Additional file 1: Table S1. of Trends in cardiovascular risk factors among U.S. men and women with and without diabetes, 1988–2014

Survey Sample Characteristics. (DOCX 25 kb

DataSheet_1_Genes and pathways correlated with heat stress responses and heat tolerance in maize kernels.docx

Global warming leads to frequent extreme weather, especially the extreme heat events, which threating the safety of maize production. Here we selected a pair of maize inbred lines, PF5411-1 and LH150, with significant differences in heat tolerance at kernel development stage. The two maize inbred lines were treated with heat stress at kernel development stage. Compared with the control groups, transcriptomic analysis identified 770 common up- and down-regulated genes between PF5411-1 and LH150 under heat stress conditions, and 41 putative TFs were predicted. Based on the interaction term of the two-factorial design, we also identified 6,744 differentially regulated genes between LH150 and PF5411-1, 111 common up-regulated and 141 common down-regulated genes were overlapped with the differentially regulated genes, respectively. Combined with proteins and metabolites data, several key pathways including seven differentially regulated genes were highly correlated with the heat tolerance of maize kernels. The first is the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway ko04141: protein processing in endoplasmic reticulum, four small heat shock protein (sHSP) genes were enriched in this pathway, participating with the process of ER-associated degradation (ERAD). The second one is the myricetin biosynthesis pathway, a differentially regulated protein, flavonoid 3’,5’-hydroxylase [EC:1.14.14.81], catalyzed the synthesis of myricetin. The third one is the raffinose metabolic pathway, one differentially regulated gene encoded the raffinose synthase controlled the synthesis of raffinose, high level of raffinose enhances the heat tolerance of maize kernels. And the last one is the ethylene signaling pathway. Taken together, our work identifies many genes responded to heat stress in maize kernels, and finds out seven genes and four pathways highly correlated with heat tolerance of maize kernels.</p

DataSheet_1_Associations of lipid parameters with non-alcoholic fatty liver disease in type 2 diabetic patients according to obesity status and metabolic goal achievement.docx

AimsNon-obese non-alcoholic fatty liver disease (NAFLD) phenotype has sparked interest and frequently occurred in type 2 diabetes mellitus (T2DM). Information on associations between lipid parameters and NAFLD in non-obese patients with diabetes has been lacking. We aimed to investigate the relationships between lipid parameters and NAFLD according to obesity status and metabolic goal achievement in T2DM patients.MethodsA total of 1,913 T2DM patients who were hospitalized between June 2018 and May 2021 were cross-sectionally assessed. We used logistic regression models to estimate the associations of lipid parameters with NAFLD risk according to obesity and metabolic goal achievement status.ResultsHigher triglycerides, non-HDL-cholesterol, and all lipid ratios including (total cholesterol/HDL-cholesterol, triglyceride/HDL-cholesterol, LDL-cholesterol/HDL-cholesterol, non-HDL-cholesterol/HDL-cholesterol), and lower HDL-cholesterol were associated with NAFLD risk in both non-obese and obese patients. The associations were stronger in non-obese patients than in obese patients. Further, the inverse associations of total cholesterol and LDL-cholesterol with NAFLD risk were only detected in non-obese patients. Triglycerides, HDL-cholesterol, and all lipid ratios studied were significantly associated with NAFLD risk, irrespective of whether the patients achieved their HbA1c, blood pressure, and LDL-cholesterol goal. The presence of poor lipids and lipid ratios were more strongly associated with NAFLD in patients who attained the HbA1c, blood pressure, and/or LDL-cholesterol goal than in those who did not achieve the goal attainment.ConclusionsThe associations of lipids and lipid ratios with NAFLD risk were stronger in T2DM patients who were non-obese and achieved the HbA1c, blood pressure, and/or LDL-cholesterol goal attainment.</p

Graphene oxide significantly inhibits cell growth at sublethal concentrations by causing extracellular iron deficiency

<p>Graphene oxide (GO)-based materials are increasingly being used in medical materials and consumer products. However, their sublethal effects on biological systems are poorly understood. Here, we report that GO (at 10 to 160 mg/L) induced significant inhibitory effects on the growth of different unicellular organisms, including eukaryotes (i.e. <i>Saccharomyces cerevisiae</i>, <i>Candida albicans</i>, and <i>Komagataella pastoris</i>) and prokaryotes (<i>Pseudomonas fluorescens</i>). Growth inhibition could not be explained by commonly reported cytotoxicity mechanisms such as plasma membrane damage or oxidative stress. Based on transcriptomic analysis and measurement of extra- and intracellular iron concentrations, we show that the inhibitory effect of GO was mainly attributable to iron deficiency caused by binding to the O-functional groups of GO, which sequestered iron and disrupted iron-related physiological and metabolic processes. This inhibitory mechanism was corroborated with supplementary experiments, where adding bathophenanthroline disulfonate—an iron chelating agent—to the culture medium exerted similar inhibition, whereas removing surface O-functional groups of GO decreased iron sequestration and significantly alleviated the inhibitory effect. These findings highlight a potential indirect detrimental effect of nanomaterials (i.e. scavenging of critical nutrients), and encourage research on potential biomedical applications of GO-based materials to sequester iron and enhance treatment of iron-dependent diseases such as cancer and some pathogenic infections.</p

Additional file 1: Table S1. of The triglyceride and glucose index (TyG) is an effective biomarker to identify nonalcoholic fatty liver disease

Odds ratios for NAFLD in different quartiles of TyG index or ALT in ALT <40 and ≥40 U/L groups. Table S2 Diagnostic value of TyG and ALT for NAFLD in ALT <40 and ≥40 U/L groups. (DOCX 19 kb

Targeting the Subpocket Enables the Discovery of Thiadiazole–Pyridazine Derivatives as Glutaminase C Inhibitors

As glutaminase C (GAC) has become an attractive target for cancer treatment by regulating glutaminolysis, thus, interest in GAC inhibitors has risen in recent years. Herein, a potential binding subpocket comprising basic residues was identified, and through extensive structure–activity relationship studies, promising inhibitors 11 and 39 were identified with robust GAC inhibitory activity and A549 cell antiproliferative activity. X-ray crystallography of the 11–GAC and 27–GAC complexes revealed a novel binding mode against GAC. The potency of 11 and 27 against GACK320A further highlighted the importance of the binding. Notably, compounds 11 and 39 regulated the cellular metabolite, thereby increasing reactive oxygen species by blocking glutamine metabolism. Compound 11 also exhibited excellent antiproliferative activity in the A549 cell xenograft model. We further proved that 11 is a safe GAC allosteric inhibitor. A basic subpocket is proposed that might provide new strategies for the development of novel GAC inhibitors in the future

Analysis of Complexes Containing Pre-miRNA Processing Activity, Dcr-1, and Loqs

<div><p>(A) S2 cell lysate was fractionated by gel filtration chromatography and analyzed for pre-<i>let-7</i> processing activity, and Dcr-1, Dcr-2, and Loqs proteins.</p> <p>(B) The sizes of the distinct complexes containing Loqs (∼630 kDa), Dcr-1 (∼480 kDa), and Dcr-2 (∼230 kDa) and the broad complex containing pre-miRNA processing activity (∼525 kDa) were estimated using molecular weight standards (thyroglobulin, 669 kDa; ferritin, 440 kDa; catalase, 232 kDa; aldolase, 158 kDa; bovine serum albumin, 67 kDa; ovalbumin, 43 kDa; chymotrypsinogen A, 25 kDa) and recombinant Dcr-2 and R2D2 proteins (rDcr-2 and rR2D2). The blue asterisk denotes the peak of pre-<i>let-7</i> processing activity detected in (A).</p> <p>(C) Fractions containing the Dcr-1 peak were pooled and immunoprecipitated with either anti-Dcr-1 or anti-Loqs antibodies. Western blotting with anti-Dcr-1 and anti-Loqs antibodies demonstrated that Dcr-1 and Loqs remained associated through gel filtration chromatography.</p></div

Silencing of a miRNA-Responsive YFP Reporter Requires <i>loqs</i> but Not <i>r2d2</i>

<div><p>(A) A YFP transgene expressed from the Pax6-promoter showed strong fluorescence in the eye and weaker fluorescence in the antennae. Due to the underlying normal red eye pigment, the YFP fluorescence was observed in only those ommatidia that are aligned with the optical axis of the stereomicroscope. In heterozygous <i>loqs</i>^f00791/CyO flies bearing a miR-277-responsive, Pax6-promotor-driven, YFP transgene, YFP fluorescence was visible in the antennae but was repressed in the eye. In contrast, in homozygous mutant <i>loqs</i>^f00791 flies, YFP fluorescence was readily detected in the eye. A strong mutation in <i>r2d2</i> did not comparably alter repression of the miR-277-regulated YFP reporter. The exposure time for the unregulated YFP reporter strain was one-fourth that used for the miR-277-responsive YFP strain. The exposure times were identical for the heterozygous and homozygous <i>loqs</i> and <i>r2d2</i> flies.</p> <p>(B) Additional images of eyes from <i>loqs</i>^f00791 heterozygous and homozygous flies bearing the miR-277-responsive YFP reporter transgene diagrammed in (A).</p> <p>(C) Quantification of fluorescence of the miR-277-responsive YFP transgene in eyes heterozygous or homozygous for <i>loqs</i> or <i>r2d2</i>. The maximum pixel intensity was measured for each eye (excluding antennae and other tissues where miR-277 does not appear to function). The graph displays the average (<i>n</i> = 13) maximum pixel intensity ± standard deviation for each homozygous genotype, normalized to the average value for the corresponding heterozygotes. Statistical significance was estimated using a two-sample Student's <i>t</i>-test assuming unequal variance.</p> <p>The images in (A) were acquired using a sensitive, GFP long-pass filter set that transmits yellow and red autofluorescence. Images in (B) and for quantitative analysis were acquired using a YFP-specific band-pass filter set that reduced the autofluorescence recorded.</p></div

Loqs Is Associated with Pre-miRNA Processing Activity in S2 Cells

<div><p>(A) Pre-miRNA processing activity co-immunoprecipitates with myc-tagged Loqs PB and with endogenous Dcr-1 or endogenous Loqs, but not with myc-tagged GFP.</p> <p>(B) Pre-miRNA processing activity co-purifies by immunoprecipitation with both Loqs protein isoforms that interact with Dcr-1, Loqs PA, and Loqs PB. The extracts used in (A) and (B) were independently prepared.</p></div

loqs^f00791 Fail to Maintain Germ-Line Stem Cells

<div><p>(A) Wild-type ovarioles contain a germarium and a developmentally ordered array of six to eight egg chambers, whereas <i>loqs</i>^f00791 mutant ovarioles contain a smaller than normal germarium, two or three pre-vitellogenic egg chambers, and a late-stage egg chamber. Wild-type and <i>loqs</i> ovarioles are shown at the same magnification.</p> <p>(B) In wild-type ovarioles, the germarium contains several newly formed germ-line cysts surrounded by somatic follicle cells. In contrast, <i>loqs</i>^f00791 mutant germaria contain few germ-line cells, which are not organized into distinct cysts. The follicle cell layer is also significantly reduced in<i>loqs</i>^f00791 germaria.</p> <p>(C) Wild-type and <i>loqs</i> mutant germaria labeled for α-Spectrin (green) and filamentous Actin (red). In wild type, anti-α-Spectrin labels the spectrosome (ss), a structure characteristic of germ-line stem cells, which are normally found at the anterior of the germarium, apposed to the somatic terminal cells (tc). The cystoblasts, the daughters of the stem cells, also contain a spectrosome, but are located posterior to the stem cells. In <i>loqs</i> mutant ovaries, spectrosome-containing cells were not detected, indicating that normal germ-line stem cells are not present. These observations indicate that stem cells are not maintained.</p> <p>In (A) and (B), ovaries were labeled for filamentous actin (red) using rhodamine phalloidin, DNA (blue) using TOTO3 (Molecular Probes), and the germ-line marker Vasa (green) using rabbit anti-Vasa antibody detected with fluorescein-conjugated anti-rabbit secondary antibody. In (B) and (C), wild-type and <i>loqs</i> germaria are shown at the same magnification.</p></div