Introgressive hybridization between members of Odocoileus was examined using the mitochondrial cytochrome-b (maternal marker) and paternal sex-determining region Y ( Sry) genes. Eight out of 130 ...free-ranging individuals from the Panhandle and Trans-Pecos regions of Texas were determined to possess the mitochondrial haplotype of mule deer ( Odocoileus hemionus (Rafinesque, 1817)) and the paternal haplotype of white-tailed deer ( Odocoileus virginianus Rafinesque, 1832). Results indicated that hybridization between deer species in Texas (6.15%) was more broadly distributed than previously reported. Previous studies demonstrated that ancient hybridization events (1.32 mya) involved the capture of the white-tailed deer mitochondrial genome by mule deer, indicating a male mule deer × a female white-tailed deer directionality relevant to hybridization. Alternatively, contemporary hybridization events indicated a reversal in directionality and suggested a cross between a female mule deer × a male white-tailed deer. The Sry gene and species assignment based on morphological characters consistently were in agreement. Further, phylogenetic relationships between Odocoileus virginianus couesi and Odocoileus hemionus eremicus warrant additional investigation as recent hybridization (>200 years) may be a mechanism that allowed these two subspecies to evolve a unique evolutionary trajectory.
The data combine objectively measured sleep and thrice‐daily salivary cortisol collected from a 4‐day diary study in a large Midwestern city with location data on all violent crimes recorded during ...the same time period for N = 82 children (Mage = 14.90, range = 11.27–18.11). The primary empirical strategy uses a within‐person design to measure the change in sleep and cortisol from the person's typical pattern on the night/day immediately following a local violent crime. On the night following a violent crime, children have later bedtimes. Children also have disrupted cortisol patterns the following morning. Supplementary analyses using varying distances of the crime to the child's home address confirm more proximate crimes correspond to later bedtimes.
Summary
Adolescence is a sensitive period for changes in both sleep and affect. Although past research has assessed the association between affect and sleep among adolescents, few studies have ...examined both trait (typical) and day‐to‐day changes in affect, and fewer still have specifically examined negative social evaluative emotions (e.g. embarrassment) in relation to sleep. Both between‐ and within‐person variations in daily affect were examined in relation to four objectively‐measured sleep outcomes (sleep hours; sleep latency; sleep efficiency; and length of wake bouts) among adolescents. Participants (N = 77 high‐school students; 42.9% female; M = 14.37 years) wore an actiwatch and completed daily‐diaries for 3 days. The results of hierarchical linear models (controlling for age, gender, race, ethnicity, parental employment status, income, puberty and caffeine) indicated that negative social evaluative emotions and high‐arousal affective experiences generally predicted poor sleep outcomes, whereas low‐arousal affective experiences were associated with good sleep outcomes. Specifically, at the person level, adolescents reporting higher negative social evaluative emotions had shorter average sleep hours, and those experiencing higher anxiety–nervousness had longer wake bouts. In addition, individuals experiencing more dysphoria (sad, depressed, lonely) had longer average sleep hours and shorter wake bouts, while those experiencing more calmness had shorter sleep latencies. At the within‐person level, individuals had longer sleep latencies following days that they had experienced high‐arousal positive affect (e.g. excitement), and had longer wake bouts following days they had experienced more negative social evaluative emotions. The results highlight the detrimental effects of negative social evaluative emotions and high‐arousal affective states for adolescent sleep.
The effects of methionine restriction (MR) in rodents are well established; it leads to decreased body and fat mass, improved glucose homeostasis and extended lifespan, despite increased energy ...intake. Leucine restriction (LR) replicates some, but not all, of these effects of MR. To determine any differences in metabolic effects between MR and LR, this study compared 8 weeks of MR (80% restriction), LR (80% restriction) and control diet in 10-month-old C57BL/6J male mice. Body composition, food intake and glucose homeostasis were measured throughout the study and biochemical analyses of white adipose tissue (WAT) and liver were performed. MR and LR decreased body and fat mass, increased food intake, elevated lipid cycling in WAT and improved whole-body glucose metabolism and hepatic insulin sensitivity in comparison to the control diet. MR produced more substantial effects than LR on body mass and glucose homeostasis and reduced hepatic lipogenic gene expression, which was absent with the LR diet. This could be a result of amino acid-specific pathways in the liver responsible for FGF21 stimulation (causing varied levels of FGF21 induction) and Akt activation. In summary, LR is effective at improving metabolic health; however, MR produces stronger effects, suggesting they activate distinct signalling pathways.
Summary
Methionine restriction (MR) decreases body weight and adiposity and improves glucose homeostasis in rodents. Similar to caloric restriction, MR extends lifespan, but is accompanied by ...increased food intake and energy expenditure. Most studies have examined MR in young animals; therefore, the aim of this study was to investigate the ability of MR to reverse age‐induced obesity and insulin resistance in adult animals. Male C57BL/6J mice aged 2 and 12 months old were fed MR (0.172% methionine) or control diet (0.86% methionine) for 8 weeks or 48 h. Food intake and whole‐body physiology were assessed and serum/tissues analyzed biochemically. Methionine restriction in 12‐month‐old mice completely reversed age‐induced alterations in body weight, adiposity, physical activity, and glucose tolerance to the levels measured in healthy 2‐month‐old control‐fed mice. This was despite a significant increase in food intake in 12‐month‐old MR‐fed mice. Methionine restriction decreased hepatic lipogenic gene expression and caused a remodeling of lipid metabolism in white adipose tissue, alongside increased insulin‐induced phosphorylation of the insulin receptor (IR) and Akt in peripheral tissues. Mice restricted of methionine exhibited increased circulating and hepatic gene expression levels of FGF21, phosphorylation of eIF2a, and expression of ATF4, with a concomitant decrease in IRE1α phosphorylation. Short‐term 48‐h MR treatment increased hepatic FGF21 expression/secretion and insulin signaling and improved whole‐body glucose homeostasis without affecting body weight. Our findings suggest that MR feeding can reverse the negative effects of aging on body mass, adiposity, and insulin resistance through an FGF21 mechanism. These findings implicate MR dietary intervention as a viable therapy for age‐induced metabolic syndrome in adult humans.
Advances in genomic analyses enable the identification of new proteins that are associated with disease. To validate these targets, tool molecules are required to demonstrate that a ligand can have a ...disease‐modifying effect. Currently, as tools are reported for only a fraction of the proteome, platforms for ligand discovery are essential to leverage insights from genomic analyses. Fragment screening offers an efficient approach to explore chemical space. Presented here is a fragment‐screening platform, termed PhABits (PhotoAffinity Bits), which utilizes a library of photoreactive fragments to covalently capture fragment–protein interactions. Hits can be profiled to determine potency and the site of crosslinking, and subsequently developed as reporters in a competitive displacement assay to identify novel hit matter. The PhABit platform is envisioned to be widely applicable to novel protein targets, identifying starting points in the development of therapeutics.
PhotoAffinity Bit (PhABit) is a photoreactive fragment‐screening platform to covalently capture fragment–protein interactions. Hits can be profiled and subsequently developed as reporters in a competitive displacement assay to identify novel hit matter. The PhABit platform is widely applicable to novel protein targets, identifying starting points in the development of therapeutics.
Large-scale human genetic data
have shown that cancer mutations display strong tissue-selectivity, but how this selectivity arises remains unclear. Here, using experimental models, functional ...genomics and analyses of patient samples, we demonstrate that the lineage transcription factor paired box 8 (PAX8) is required for oncogenic signalling by two common genetic alterations that cause clear cell renal cell carcinoma (ccRCC) in humans: the germline variant rs7948643 at 11q13.3 and somatic inactivation of the von Hippel-Lindau tumour suppressor (VHL)
. VHL loss, which is observed in about 90% of ccRCCs, can lead to hypoxia-inducible factor 2α (HIF2A) stabilization
. We show that HIF2A is preferentially recruited to PAX8-bound transcriptional enhancers, including a pro-tumorigenic cyclin D1 (CCND1) enhancer that is controlled by PAX8 and HIF2A. The ccRCC-protective allele C at rs7948643 inhibits PAX8 binding at this enhancer and downstream activation of CCND1 expression. Co-option of a PAX8-dependent physiological programme that supports the proliferation of normal renal epithelial cells is also required for MYC expression from the ccRCC metastasis-associated amplicons at 8q21.3-q24.3 (ref.
). These results demonstrate that transcriptional lineage factors are essential for oncogenic signalling and that they mediate tissue-specific cancer risk associated with somatic and inherited genetic variants.
Carboxylesterases (CEs) are a class of enzymes that catalyze the hydrolysis of esters in a variety of endogenous and exogenous molecules. CEs play an important role in drug metabolism, in the onset ...and progression of disease, and can be harnessed for prodrug activation strategies. As such, the regulation of CEs is an important clinical and pharmaceutical consideration. Here, we report the first ratiometric sensor for CE activity using Raman spectroscopy based on a bisarylbutadiyne scaffold. The sensor was shown to be highly sensitive and specific for CE detection and had low cellular cytotoxicity. In hepatocyte cells, the ratiometric detection of esterase activity was possible, and the result was validated by multimodal imaging with standard viability stains used for fluorescence microscopy within the same cell population. In addition, we show that the detection of localized ultraviolet damage in a mixed cell population was possible using stimulated Raman scattering microscopy coupled with spectral phasor analysis. This sensor demonstrates the practical advantages of low molecular weight sensors that are detected using ratiometric Raman imaging and will have applications in drug discovery and biomedical research.
Methods for rapid identification of chemical tools are essential for the validation of emerging targets and to provide medicinal chemistry starting points for the development of new medicines. Here, ...we report a screening platform that combines 'direct-to-biology' high-throughput chemistry (D2B-HTC) with photoreactive fragments. The platform enabled the rapid synthesis of >1000 PhotoAffinity Bits (HTC-PhABits) in 384-well plates in 24 h and their subsequent screening as crude reaction products with a protein target without purification. Screening the HTC-PhABit library with carbonic anhydrase I (CAI) afforded 7 hits (0.7% hit rate), which were found to covalently crosslink in the Zn
2+
binding pocket. A powerful advantage of the D2B-HTC screening platform is the ability to rapidly perform iterative design-make-test cycles, accelerating the development and optimisation of chemical tools and medicinal chemistry starting points with little investment of resource.
A photoreactive fragment screening platform employing direct-to-biology high-throughput chemistry (D2B-HTC) for the rapid iterative synthesis and screening of libraries of photoaffinity bits.
Protein tyrosine phosphatase-1B (PTP1B) negatively regulates insulin and leptin signaling, rendering it an attractive drug target for treatment of obesity-induced insulin resistance. However, some ...studies suggest caution when targeting macrophage PTP1B, due to its potential anti-inflammatory role. We assessed the role of macrophage PTP1B in inflammation and whole-body metabolism using myeloid-cell (LysM) PTP1B knockout mice (LysM PTP1B). LysM PTP1B mice were protected against lipopolysaccharide (LPS)-induced endotoxemia and hepatic damage associated with decreased proinflammatory cytokine secretion in vivo. In vitro, LPS-treated LysM PTP1B bone marrow-derived macrophages (BMDMs) displayed increased interleukin (IL)-10 mRNA expression, with a concomitant decrease in TNF-α mRNA levels. These anti-inflammatory effects were associated with increased LPS- and IL-10-induced STAT3 phosphorylation in LysM PTP1B BMDMs. Chronic inflammation induced by high-fat (HF) feeding led to equally beneficial effects of macrophage PTP1B deficiency; LysM PTP1B mice exhibited improved glucose and insulin tolerance, protection against LPS-induced hyperinsulinemia, decreased macrophage infiltration into adipose tissue, and decreased liver damage. HF-fed LysM PTP1B mice had increased basal and LPS-induced IL-10 levels, associated with elevated STAT3 phosphorylation in splenic cells, IL-10 mRNA expression, and expansion of cells expressing myeloid markers. These increased IL-10 levels negatively correlated with circulating insulin and alanine transferase levels. Our studies implicate myeloid PTP1B in negative regulation of STAT3/IL-10-mediated signaling, highlighting its inhibition as a potential anti-inflammatory and antidiabetic target in obesity.
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