Meclofenamic acid selectively inhibits FTO demethylation of m6A over ALKBH5

Two human demethylases, the fat mass and obesity-associated (FTO) enzyme and ALKBH5, oxidatively demethylate abundant N⁶-methyladenosine (m⁶A) residues in mRNA. Achieving a method for selective inhibition of FTO over ALKBH5 remains a challenge, however. Here, we have identified meclofenamic acid (MA) as a highly selective inhibitor of FTO. MA is a non-steroidal, anti-inflammatory drug that mechanistic studies indicate competes with FTO binding for the m⁶A-containing nucleic acid. The structure of FTO/MA has revealed much about the inhibitory function of FTO. Our newfound understanding, revealed herein, of the part of the nucleotide recognition lid (NRL) in FTO, for example, has helped elucidate the principles behind the selectivity of FTO over ALKBH5. Treatment of HeLa cells with the ethyl ester form of MA (MA2) has led to elevated levels of m⁶A modification in mRNA. Our collective results highlight the development of functional probes of the FTO enzyme that will (i) enable future biological studies and (ii) pave the way for the rational design of potent and specific inhibitors of FTO for use in medicine.     

Correlations of fecal bacterial communities with age and living region for the elderly living in Bama,Guangxi, China

Bama County (Guangxi, China) is famous for its longevous population. In this study, intestinal microflora of 17 healthy elderly subjects of different ages and from different regions (rural and urban) in Bama, were analyzed by denaturing gradient gel electrophoresis (DGGE). Significant effects of age and living region on the whole intestinal bacterial communities were observed by redundancy analysis (RDA). A total of 11 bacterial strains that were correlated with age and living region were identified using a t-value biplot combined with band sequencing. Four bacterial strains were correlated with both age and living region of the elderly in Bama. Two Bacteroides strains and one Ruminococcaceae strain were abundant in the rural, younger elderly; conversely, one Desulfovibrio strain was high in the urban, older elderly. Another Bacteroidetes strain was only correlated with the participant’s age, and its abundance increased with the age of the elderly. The richness of one Clostridium sordellii strain, which was only correlated with the elderly living region, was high in the urban elderly. The study also found five other novel bacterial strains that were correlated with the age or living region of the elderly in Bama. These results expand our understanding of age- and region-effects on the intestinal microflora of the elderly and raise the possibility of developing probiotics originating from centenarians.     

Combination Therapy with the Histone Deacetylase Inhibitor LBH589 and Radiation Is an Effective Regimen for Prostate Cancer Cells

Radiation therapy (RT) continues to be one of the most popular treatment options for localized prostate cancer (CaP). The purpose of the study was to investigate the in vitro effect of LBH589 alone and in combination with RT on the growth and survival of CaP cell lines and the possible mechanisms of radiosensitization of this combination therapy. The effect of LBH589 alone or in combination with RT on two CaP cell lines (PC-3 and LNCaP) and a normal prostatic epithelial cell line (RWPE-1) was studied by MTT and clonogenic assays, cell cycle analysis, western blotting of apoptosis-related and cell check point proteins, and DNA double strand break (DSB) repair markers. The immunofluorescence staining was used to further confirm DSB expression in treated CaP cells. Our results indicate that LBH589 inhibited proliferation in both CaP and normal prostatic epithelial cells in a time-and-dose-dependent manner; low-dose of LBH589 (IC₂₀) combined with RT greatly improved efficiency of cell killing in CaP cells; compared to RT alone, the combination treatment with LBH589 and RT induced more apoptosis and led to a steady increase of sub-G1 population and abolishment of RT-induced G2/M arrest, increased and persistent DSB, less activation of non-homologous end joining (NHEJ)/homologous recombination (HR) repair pathways and a panel of cell cycle related proteins. These results suggest that LBH589 is a potential agent to increase radiosensitivity of human CaP cells. LBH589 used either alone, or in combination with RT is an attractive strategy for treating human CaP.     

Structure and Composition of Natural Gmelin Larch (Larix gmelinii var. gmelinii) Forests in Response to Spatial Climatic Changes

Background

Many theoretical researches predicted that the larch species would decrease drastically in China under future climatic changes. However, responses of the structural and compositional changes of Gmelin larch (Larix gmelinii var. gmelinii) forests to climatic changes have rarely been reported.

Methodology/Principal Findings

Field survey was conducted to examine the structures and compositions of natural Gmelin larch forests along a climatic gradient. Stepwise linear regression analyses incorporating linear and quadratic components of climatic and non-climatic factors were performed on the structural and compositional attributes of those natural Gmelin larch forests. Isothermality, Max Temperature of Warmest Month (TempWarmestMonth), Precipitation of Wettest Month (PrecipWettestMonth), Precipitation Seasonality (PrecipSeasonality) and Precipitation of Driest Quarter (PrecipDriestQuarter) were observed to be effective climatic factors in controlling structure and composition of Gmelin larch forests. Isothermality significantly affected total basal area of larch, while TempWarmestMonth, PrecipWettestMonth and PrecipSeasonality significantly affected total basal area of Mongolian pine, and PrecipDriestQuarter significantly affected mean DBH of larch, stand density of larch and total basal area of spruce and fir.

Conclusions/Significance

The summer and winter temperatures and precipitations are all predicted to increase in future in Northeast China. Our results showed the increase of total basal area of spruce and fir, the suppression of regeneration and the decrease of stand density of larch under increased winter precipitation, and the decrease of total basal area of larch under increased summer temperature in the region of current Gmelin larch forest. Therefore, we suggest that larch would decrease and spruce and fir would increase in the region of future Gmelin larch forest.     

Synthesis and biological evaluation of novel F-18 labeled pyrazolo[1,5-a]pyrimidine derivatives: potential PET imaging agents for tumor detection

Two novel pyrazolo[1,5-a]pyrimidine derivatives, 7-(2-[¹⁸F]fluoroethylamino)-5-methylpyrazolo[1,5-a]pyrimidine-3-carbonitrile ([¹⁸F]FEMPPC, [¹⁸F]1) and N-(2-(3-cyano-5-methylpyrazolo[1,5-a]pyrimidin-7-ylamino)ethyl)-2-[¹⁸F]fluoro-4-nitrobenzamide ([¹⁸F]FCMPPN, [¹⁸F]2), have been designed and successively labeled with ¹⁸F by the nucleophilic substitution employing tosylate and nitryl as leaving groups, respectively. The radiochemical synthesis of both compounds was completed within 60 min with final high-performance liquid chromatography purification included. The corresponding radiochemical yields (without decay correction) were approximately 35% and 30%, respectively. Meanwhile, we compared the uptake characteristics of [¹⁸F]1 and [¹⁸F]2 with those of [¹⁸F]FDG and L-[¹⁸F]FET in S180 tumor cells. Furthermore, the tumor uptake of [¹⁸F]1 and [¹⁸F]2 was assessed in mice bearing S180 tumor and compared with [¹⁸F]FDG and L-[¹⁸F]FET in the same animal model. In vitro cell uptake studies showed [¹⁸F]1 had higher uptake than [¹⁸F]FDG, [¹⁸F]2 and L-[¹⁸F]FET over the 2 h period. In ex vivo biodistribution showed tumor/brain uptake ratios of [¹⁸F]2 were 12.35, 10.44, 8.69 and 5.13 at 15 min, 30 min, 60 min and 120 min post-injection, much higher than those of L-[¹⁸F]FET (2.43, 2.54, 2.93 and 2.95) and [¹⁸F]FDG (0.59, 0.61, 1.02 and 1.33) at the same time point. What’s more, the uptake of [¹⁸F]1 in tumor was 1.88, 4.37, 5.51, 2.95 and 2.88 at 5 min, 15 min, 30 min, 60 min and 120 min post-injection, respectively. There was a remarkable increasing trend before 30 min. The same trend was present for L-[¹⁸F]FET before 30 min and [¹⁸F]FDG before 60 min. Additionally, the tumor/brain uptake ratios of [¹⁸F]1 were superior to those of [¹⁸F]FDG at all the selected time points, the tumor/muscle and tumor/blood uptake ratios of [¹⁸F]1 at 30 min were higher than those of L-[¹⁸F]FET at the same time point. MicroPET image of [¹⁸F]1 administered into S180 tumor-bearing mouse acquired at 30 min post-injection illustrated that the uptake in S180 tumor was obvious. These results suggest that compound [¹⁸F]1 could be a new probe for PET tumor imaging.     

A heat shock protein 90 binding domain in endothelial nitric-oxide synthase influences enzyme function

Previous reports suggest heat shock protein 90 (hsp90) associates with endothelial nitric-oxide synthase (eNOS) to increase nitric oxide (*NO) generation. Ansamycin inhibition of chaperone-dependent activity increases eNOS generation of superoxide anion (O(2)(*)) upon enzyme activation. In the present study we identify where hsp90 binds to eNOS using overlapping decoy peptides based on the amino acid (aa) sequence of eNOS (291-420). B1, B2, and B3 peptides inhibited hsp90 association with eNOS in cell lysates from proliferating bovine aortic endothelial cells. B2 (aa 301-320), common to both B1 and B3, decreased stimulated *NO production and hsp90 association in bovine aortic endothelial cells. The B2/B3 peptide was redesigned to TSB2 that includes a TAT protein transduction domain and shortened to 14 aa. TSB2 impaired vasodilation of isolated facialis arteries in vitro and in vivo and increased eNOS-dependent O(2)(*) generation in native endothelial cells on mouse aortas, whereas a control peptide, TSB(Ctr), which has the four glutamic acids in TSB2 substituted with alanine, showed no such effects. Site-directed mutagenesis of eNOS at 310, 314, 318, and 323 Glu to Ala yields an eNOS mutant that exhibited reduced hsp90 association and generated O(2)(*) rather than *NO upon activation. Together, these data demonstrate that hsp90 associates with eNOS at aa 310-323. Moreover, a decoy peptide based on this sequence is sufficient to displace hsp90 from eNOS and uncouple eNOS activity from *NO generation. Thus, Glu-310, Glu-314, Glu-318, and Glu-323 in eNOS, although each does not do much by itself, synergistically they increase "cooperativity" in the association step that is critical for maintaining hsp90-eNOS interactions and promoting coupled eNOS activity. Such chaperone-dependent signaling may play an important role in modulating the balance of *NO and O(2)(*) generation from eNOS and, therefore, vascular function.     

64层螺旋CT三维重建技术对下肢动脉粥样硬化病变的显示

目的探讨64层螺旋CT三维重建技术对下肢动脉病变的显示。方法103例下肢动脉粥样硬化的患者行64层螺旋CT血管造影检查资料在工作站进行多种三维重建,分析其影像表现。结果103例患者均获得了成功的造影图像,2459根下肢动脉血管得到了显示。其中845根(34.4%)下肢动脉血管正常;672根(27.3%)轻度狭窄;327根(13.3%)中度狭窄;292根(11.9%)重度狭窄,323根(13.1%)下肢动脉血管闭塞。结论64层螺旋CT三维重建是又一种无创性的、可靠的、简便的检查下肢动脉粥样硬化的方法,具有较高的临床价值。     

Knockdown of PSMC3IP suppresses the proliferation and xenografted tumorigenesis of hepatocellular carcinoma cell

Hepatocellular carcinoma (HCC) is the fifth most frequent malignancy and the second leading cause of cancer-related death worldwide. Proteasome 26S subunit ATPase 3 interacting protein (PSMC3IP) is an oncogene in breast cancer, while its role in HCC remains unclear. Here, we found that PSMC3IP was critical for the cell proliferation and tumorigenic capacity of HCC cells. Upregulation of PSMC3IP was observed in HCC specimens, and high PSMC3IP expression predicted poor overall survival of HCC patients. In vitro, knockdown of PSMC3IP blunted the proliferation and colony formation of BEL-7404 and SMMC-7721 cells. Likewise, PSMC3IP silencing suppressed the xenografted tumor development of BEL-7404 cells. Mechanistically, apoptosis was enhanced after PSMC3IP knockdown in both BEL-7404 and SMMC-7721 cells. At the molecular level, TP53 and GNG4 were upregulated and eukaryotic translation initiation factor 4E (EIF4E) and insulin like growth factor 1 receptor (IGF1R) were downregulated in shPSMC3IP compared with shCtrl BEL-7404 cells. Therefore, targeting PSMC3IP maybe a promising strategy for HCC.     

Remarkably Enhanced Hydrogen Generation of Organolead Halide Perovskites via Piezocatalysis and Photocatalysis

To alleviate photoinduced charge recombination in semiconducting nanomaterials represents an important endeavor toward high‐efficiency photocatalysis. Here a judicious integration of piezoelectric and photocatalytic properties of organolead halide perovskite CH₃NH₃PbI₃ (MAPbI₃) to enable a piezophotocatalytic activity under simultaneous ultrasonication and visible light illumination for markedly enhanced photocatalytic hydrogen generation of MAPbI₃ is reported. The conduction band minimum of MAPbI₃ is higher than hydrogen generation potential (0.046 V vs normal hydrogen electrode), thereby rendering efficient hydrogen evolution. In addition, the noncentrosymmetric crystal structure of MAPbI₃ enables its piezoelectric properties. Thus, MAPbI₃ readily responds to external mechanical force, creating a built‐in electric field for collective piezophotocatalysis as a result of effective separation of photogenerated charge carriers. The experimental results show that MAPbI₃ powders exhibit superior piezophotocatalytic hydrogen generation rate (23.30 µmol h^?1) in hydroiodic acid (HI) solution upon concurrent light and mechanical stimulations, much higher than that of piezocatalytic (i.e., 2.21 µmol h^?1) and photocatalytic (i.e., 3.42 µmol h^?1) hydrogen evolution rate as well as their sum (i.e., 5.63 µmol h^?1). The piezophotocatalytic strategy provides a new way to control the recombination of photoinduced charge carriers by cooperatively capitalizing on piezocatalysis and photocatalysis of organolead halide perovskites to yield highly efficient piezophotocatalysis.