Search

Objective: To explore the value of multi-voxel-based monomer 1H-MRS (magnetic resonance imaging) in the classification of gliomas. Methods: 26 gliomas confirmed by surgery and pathology were examined by conventional MRI, multi-voxel 2D, and monomer 1H-MRS before surgery. Three types of regions of interest (ROI) were used to measure Cho/NAA (Choline hydroxide solution/N-acetyl aspartic acid) values in the tumor. Methods: In the multi-voxel 2D hydrogen proton spectrum, the solid part of the tumor or the most intensive region of the tumor was selected to determine the Cho/NAA value. At the same time, in the multi-voxel 2D hydrogen proton spectrum, the Fun/tool spectra's copy-2DBmin analysis software was used to obtain the Cho/NAA value distribution. Anatomically superimposed the pseudo-color image, and measure the Cho/NAA value in its largest Cho/NAA value region. In addition, based on the level set algorithm, it uses the Otsu algorithm to initially locate the edema area and provide the initial contour. Results: The Cho/NAA values measured by the ROI selection method were statistically significant, and the sensitivity and specificity of judging the benign and malignant gliomas indicated that Method 3 had the highest diagnostic value. Conclusion: Multi-voxel-based monomeric 1H-MRS magnetic resonance imaging has the highest value in guiding glioma classification and can improve the accuracy of glioma classification.

Strain Q3-56T, isolated from Arctic tundra soil, was found to be a Gram-stain-negative, yellow-pigmented, oxidase- and catalase-positive, non-motile, non-spore-forming, rod-shaped and aerobic bacterium. Strain Q3-56T grew optimally at pH 7.0 and 28 °C. The strain could tolerate up to 1 % (w/v) NaCl with optimum growth in the absence of NaCl. The strain was not sensitive to oxacillin and ceftazidime. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain Q3-56T belonged to the genus Dyadobacter . Strain Q3-56T showed the highest sequence similarities to Dyadobacter luticola T17T (96.58 %), Dyadobacter ginsengisoli Gsoil 043T (96.50 %), Dyadobacter flavalbus NS28T (96.43 %) and Dyadobacter bucti QTA69T (96.43 %). The predominant respiratory isoprenoid quinone was identified as MK-7, The polar lipid profile of strain Q3-56T was found to contain one phosphatidylethanolamine, three unidentified aminolipids, three unidentified lipids and one unidentified phospholipid. The G+C content of the genomic DNA was determined to be 49.1 mol%. The main fatty acids were summed feature 3 (comprising C16 : 1 ω7c/C16 : 1 ω6c), iso-C15 : 0, C16 : 1 ω5c and iso-C16 : 1 3-OH. On the basis of the evidence presented in this study, a novel species of the genus Dyadobacter , Dyadobacter sandarakinus sp. nov., is proposed, with the type strain Q3-56T (=CCTCC AB 2019271T=KCTC 72739T). Emended descriptions of Dyadobacter alkalitolerans , Dyadobacter koreensis and Dyadobacter psychrophilus are also provided.

Objective: Septic shock is the severe complication of sepsis, with a high mortality. The inflammatory response regulates the immune status and mediates the progression of septic shock. In this study, we aim to identify the key immune-related genes (IRGs) of septic shock and explore their potential mechanism.Methods: Gene expression profiles of septic shock blood samples and normal whole blood samples were retrieved from the Gene Expression Omnibus (GEO) and Genotype-Tissue Expression Portal (GTEx). The differential expression genes (DEGs) and septic shock-specific immune-related genes (SSSIRGs) were evaluated and identified, along with the immune components by “cell type identification by estimating relative subsets of RNA transcripts (CIBERSORT, version x)” algorithm. Additionally, in order to explore the key regulatory network, the relationship among SSSIRGs, upstream transcription factors (TFs), and downstream signaling pathways were also identified by Gene Set Variation Analysis (GSVA) and co-expression analysis. Moreover, the Connectivity Map (CMap) analysis was applied to find bioactive small molecules against the members of regulation network while Chromatin Immunoprecipitation sequencing (ChIP-seq) and Assay for Targeting Accessible-Chromatin with high-throughput sequencing (ATAC-seq) data were used to validate the regulation mechanism of the network.Results: A total of 14,843 DEGs were found between 63 septic shock blood samples and 337 normal whole blood samples. Then, we identified septic shock-specific 839 IRGs as the intersection of DEGs and IRGs. Moreover, we uncovered the regulatory networks based on co-expression analysis and found 28 co-expression interaction pairs. In the regulation network, protein phosphatase 3, catalytic subunit, alpha isozyme (PPP3CA) may regulate late estrogen response, glycolysis and TNFα signaling via NFκB and HLA; Kirsten rat sarcoma viral oncogene homolog (KRAS) may be related to late estrogen response and HLA; and Toll-like receptor 8 (TLR8) may be associated with TNFα signaling via NFκB. And the regulation mechanisms between TFs and IRGs (TLR8, PPP3CA, and KRAS) were validated by ChIP-seq and ATAC-seq.Conclusion: Our data identify three SSSIRGs (TLR8, PPP3CA, and KRAS) as candidate therapeutic targets for septic shock and provide constructed regulatory networks in septic shock to explore its potential mechanism.

Obesity is a growing health concern, as it increases risk for heart disease, hypertension, type 2 diabetes, cancer, COVID-19 related hospitalizations and mortality. However, current weight loss therapies are often associated with psychiatric or cardiovascular side effects or poor tolerability that limit their long-term use. The hypothalamic neuropeptide, oxytocin (OT), mediates a wide range of physiologic actions, which include reproductive behavior, formation of prosocial behaviors and control of body weight. We and others have shown that OT circumvents leptin resistance and elicits weight loss in diet-induced obese rodents and non-human primates by reducing both food intake and increasing energy expenditure (EE). Chronic intranasal OT also elicits promising effects on weight loss in obese humans. This review evaluates the potential use of OT as a therapeutic strategy to treat obesity in rodents, non-human primates, and humans, and identifies potential mechanisms that mediate this effect.

Objective Bariatric surgery acutely improves glucose control, an effect that is generally sustained for years in most patients. The acute postoperative glycemic reduction is at least partially mediated by enhanced incretin secretion and islet function, and occurs independent of caloric restriction, whereas the sustained improvement in glucose control is associated with increased insulin sensitivity. However, studies in humans with bariatric surgery suggest that these elevations are not static but undergo coordinated regulation throughout the postoperative time course. The studies described here test the hypothesis that incretin secretion, islet function, and peripheral insulin sensitivity undergo temporal regulation following bariatric surgery as a means to regulate glucose homeostasis. Methods Incretin secretion, islet function, and insulin sensitivity in mice with vertical sleeve gastrectomy (VSG) were compared to sham-operated controls that were pair-fed for 90d, matching food consumption and body-weight between groups. Results Glucose clearance and insulin secretion were enhanced in VSG mice compared to controls during mixed-meal tolerance tests (MMTT) at 12 and 80 days postoperatively, as were prandial GLP-1, GIP, and glucagon levels. Insulin sensitivity was comparable between groups 14d after surgery, but significantly greater in the VSG group at day 75, despite similar body-weight gain between groups. Glucose stimulated insulin secretion was greater in VSG mice compared to controls in vivo (I.P. glucose injection) and ex vivo (islet perifusion) indicating a rapid and sustained enhancement of β-cell function after surgery. Notably, glycemia following a MMTT was progressively higher over time in the control animals but improved in the VSG mice at 80d despite weight regain. However, meal-stimulated incretin secretion decreased in VSG mice from 10 to 80 days postoperative, as did meal-stimulated and I.P. glucose-stimulated insulin secretion. This occurred over the same time period that insulin sensitivity was enhanced in VSG mice, suggesting postoperative islet output is tightly regulated by insulin demand. Conclusions These data demonstrate a dynamic, multifactorial physiology for improved glucose control after VSG, whereby rapidly elevated insulin secretion is complimented by later enhancements in insulin sensitivity. Critically, the glucose lowering effect of VSG is demonstrably larger than that of caloric-restriction, suggesting these adaptations are mediated by surgical modification of gastrointestinal anatomy and not weight-loss per se., Highlights • β-cell glucose sensitivity is enhanced 90d after VSG compared to controls, coincident with improved glucose tolerance. • Prandial GLP-1 and GIP are elevated 12d following VSG but return to preoperative levels 80d after VSG. • Insulin sensitivity is enhanced 75d after surgery, but not 14d after surgery, in mice with VSG compared to controls. • Mixed-meal glucose control is improved from 12d to 80d in VSG mice, but worsens in controls despite similar body-weight.

Black rockfish (Sebastes schlegelii) is a teleost species where eggs are fertilized internally and retained in the maternal reproductive system, where they undergo development until live birth (termed viviparity). In the present study, we report a chromosome-level black rockfish genome assembly. High-throughput transcriptome analysis (RNA-seq and ATAC-seq), coupled with in situ hybridization (ISH) and immunofluorescence, identify several candidate genes for maternal preparation, sperm storage and release, and hatching. We propose that zona pellucida (ZP) genes retain sperm at the oocyte envelope, while genes in two distinct astacin metalloproteinase subfamilies serve to release sperm from the ZP and free the embryo from chorion at pre-hatching stage. Finally, we present a model of black rockfish reproduction, and propose that the rockfish ovarian wall has a similar function to uterus of mammals. Taken together, these genomic data reveal unprecedented insights into the evolution of an unusual teleost life history strategy, and provide a sound foundation for studying viviparity in non-mammalian vertebrates and an invaluable resource for rockfish ecology and evolution research.

Exogenous ghrelin reduces glucose-stimulated insulin secretion and endogenous ghrelin protects against hypoglycemia during starvation. Islet ε-cells produce ghrelin and δ-cells express growth hormone secretagogue receptor (GHSR), suggesting the possibility of a paracrine mechanism for islet ghrelin to reach high local concentrations and affect insulin secretion. GHSR has high constitutive activity and may act independently of ghrelin. The objective in this study was to determine whether an intraislet ghrelin-GHSR axis modulates insulin secretion and glucose metabolism using mouse models lacking ghrelin (Ghrl−/−) or GHSR (Ghsr−/−). Ghsr−/− and Ghsr+/+ mice had comparable islet ghrelin concentrations. Exogenous ghrelin decreased insulin secretion in perifused isolated islets in a GHSR-dependent manner. Islets isolated from Ghrl−/− or Ghsr−/− mice did not differ from controls in glucose-, alanine-, or GLP-1–stimulated insulin secretion during perifusion. Consistent with this finding, Ghrl−/− and Ghsr−/− male mice studied after either 6 or 16 h of fasting had blood glucose concentrations comparable with those of controls following intraperitoneal glucose, or insulin tolerance tests, or after mixed nutrient meals. Collectively, our data provide strong evidence against a paracrine ghrelin-GHSR axis mediating insulin secretion or glucose tolerance in lean, chow-fed adult mice.

P450c17, a key enzyme in the steroid generation pathway, plays an important role in the production of sex steroid and cortisol. In this study, two cyp17 gene isoforms, Pocyp17-I and Pocyp17-II were isolated from Paralichthys olivaceus gonads. Domain architecture analysis of Pocyp17-I and Pocyp17-II revealed that they had three regions important to enzymatic function. Structural analysis showed that Pocyp17-I and Pocyp17-II had 8 and 9 exons respectively, and the difference was caused by the insertion of an extra intron (intron1) in the latter. Quantitative real-time polymerase chain reaction results indicated that the expression of these two genes showed sexually dimorphism that Pocyp17-I and Pocyp17-II were highest expressed in testis and ovary, respectively. The in situ hybridization analysis of gonads indicated that Pocyp17-I and Pocyp17-II mRNA were both detected in oocytes, spermatocytes and Sertoli cells. After injection of androgen and estrogen (17α-methyltestosterone, 17β-estradiol) of different concentrations, the expression level of Pocyp17-I decreased significantly (P 0.01), whereas estrogen had no influence on Pocyp17-II, but androgen upregulated the expression of Pocyp17-II (P 0.05). Moreover, Pocyp17-I expression level was down-regulated significantly by NR0b1 but up-regulated by NR5a2 (P 0.05), whereas Pocyp17-II expression level was down-regulated significantly by NR0b1 and NR5a2 (P 0.05). All these results demonstrated that there were differences in expression patterns, feedback actions of sex hormones and transcriptional regulations between cyp17-I and cyp17-II, which revealed that cyp17-I and cyp17-II might perform different functions in sex hormones biosynthesis and gonadal differentiation in Japanese flounder.

Convenient and time-saving one-step strategies for detecting ultralow concentrations of protein biomarkers play key roles in rapid disease diagnosis. In this study, we report a one-step detection method based on a nanofibrous sensing platform via the combination of proximity-induced DNA strand displacement (PiDSD), catalytic hairpin assembly (CHA) amplification and thioflavin T (ThT) binding. The interface behaviors on the nanofibrous membrane were studied to promote interface reaction kinetics and thermodynamics. Thrombin was used as a model biomarker, and the nanofibrous sensing platform achieved a limit of detection as low as 1.0 pM, a wide linear range of 50 pM to 5 nM, excellent specificity and good long-term stability. Compared with previous one-step thrombin detection methods, our one-step detection method is label-free, convenient and much more sensitive; it has potential applications for protein detection in point-to-care testing (POCT) and early diagnosis.

Dental caries is one of the most prevalent human oral diseases. The pH of the oral environment is lower than the critical pH point, which causes the demineralization of the hard tissues of teeth and finally leads to dental caries. Therefore, it is important to have an accurate, sensitive detecting assay of the critical pH. An electrospun fibrous membrane was reported to modulate immobilized bromocresol green (BCG), a common pH indicator, by shifting the acid dissociation constant (pKa) from 4.6 to 5.3. Based on spectroscopic and colorimetric characterizations, it was demonstrated that the BCG–polystyrene/polyvinylpyrrolidone (BCG–PS/PVP) electrospun fibrous membrane achieved a sensitive and fast response; moreover, its use exhibits a convenient sensing strategy for visualizing the critical pH point of 5.5 that induces tooth demineralization. Compared with a common pH meter or pH test papers, the BCG–PS/PVP electrospun fibrous membrane could distinguish pH 5.5 from its adjacent pH value with higher accuracy and much more convenience. We could also evaluate the inhibitory effect of the anti-caries agents on bacterial growth and acid production by the platform. The BCG–PS/PVP electrospun fibrous membrane is expected to have potential applications in the monitoring and prevention of early caries.

Spermatogenesis is the process during which spermatocytes undergo meiosis to produce spermatozoa. Sertoli-Sertoli and Sertoli-spermatogenic junctions are important for the maturity and release of sperms. These junctions, which are related to male infertility, have been reported in mammals but are unclear in teleost species. In this study, the RNAs in the gonads of 1.5-year-old Japanese flounders were sequenced using Illumina. The miRNA expression profiles in the mature gonads of males and females were characterized. A total of 277 mature miRNAs were screened in both gonads. Among them, 25 miRNAs were expressed differently in the ovary and testis. Afterward, miR-1388 was selected for its high expression level in the ovary. Gene ontology analysis revealed that the target genes of miR-1388 were highly involved in cell-cell adhesion. Nectin2l, the target of miR-1388, is a Ca-independent cell adhesion molecule that was demonstrated to be testis-biased via quantitative real-time PCR and located in Sertoli cells and spermatocytes via in situ hybridization. Our present study predicted the potential functions of sex-biased miRNA and indicated that miR-1388 regulates cell junctions by inhibiting nectin2l, which may have an important role in the later development of testes. Our study lays the ground work of miRNAs' function in the cell junctions of germ cells.

Objective The blood–brain barrier (BBB) regulates the entry of substrates and peptides into the brain. Ghrelin is mainly produced in the stomach but exerts its actions in the central nervous system (CNS) by crossing the BBB. Once present in the CNS, ghrelin can act in the hypothalamus to regulate food intake, in the hippocampus to regulate neurogenesis, and in the olfactory bulb to regulate food-seeking behavior. The goal of this study was to determine whether the primary signaling receptor for ghrelin, the growth hormone secretagogue receptor (GHSR), mediates the transport of ghrelin from blood to brain. Methods We utilized the sensitive and quantitative multiple-time regression analysis technique to determine the transport rate of mouse and human acyl ghrelin (AG) and desacyl ghrelin (DAG) in wildtype and Ghsr null mice. We also measured the regional distribution of these ghrelin peptides throughout the brain. Lastly, we characterized the transport characteristics of human DAG by measuring the stability in serum and brain, saturability of transport, and the complete transfer across the brain endothelial cell. Results We found the transport rate across the BBB of both forms of ghrelin, AG, and DAG, were not affected by the loss of GHSR. We did find differences in the transport rate between the two isoforms, with DAG being faster than AG; this was dependent on the species of ghrelin, human being faster than mouse. Lastly, based on the ubiquitous properties of ghrelin throughout the CNS, we looked at regional distribution of ghrelin uptake and found the highest levels of uptake in the olfactory bulb. Conclusions The data presented here suggest that ghrelin transport can occur independently of the GHSR, and ghrelin uptake varies regionally throughout the brain. These findings better our understanding of the gut-brain communication and may lead to new understandings of ghrelin physiology., Highlights • Ghrelin is transported across the BBB in the absence of GHSR. • Human DAG is transported the fastest compared to mDAG > hAG > mAG. • The olfactory bulb contained the greatest amount of ghrelin.

A plasmonic nanoplatform to perform an enzyme-free, naked-eye, and trace discrimination of single-base mutation from fully matched sequence is reported. The nanoplatform showed great potential to enhance catalytic hairpin assembly (CHA) amplification efficiency and biocatalytic activity of hemin/G-quadruplex (DNAzyme). When human immunodeficiency virus (HIV) DNA biomarker was used as the model analyst, a naked-eye detection with high selectivity and high sensitivity down to 10–17 M in whole serum was achieved by observing red-to-blue color change. Single-base mismatch and two-base mismatch were detected at the low concentrations of 10–11 and 10–8 M, respectively. The naked-eye detection based on the enzyme-free plasmonic nanoplatform is expected to have potential applications ranging from quick detection and early diagnostics to point-of-care research.

Bariatric surgeries, including vertical sleeve gastrectomy (VSG), resolve diabetes in 40–50% of patients. Studies examining the molecular mechanisms underlying this effect have centered on the role of the insulinotropic glucagon-like peptide 1 (GLP-1), in great part because of the ∼10-fold rise in its circulating levels after surgery. However, there is currently debate over the role of direct β-cell signaling by GLP-1 to mediate improved glucose tolerance following surgery. In order to assess the importance of β-cell GLP-1 receptor (GLP-1R) for improving glucose control after VSG, a mouse model of this procedure was developed and combined with a genetically modified mouse line allowing an inducible, β-cell–specific Glp1r knockdown (Glp1rβ-cell-ko). Mice with VSG lost ∼20% of body weight over 30 days compared with sham-operated controls and had a ∼60% improvement in glucose tolerance. Isolated islets from VSG mice had significantly greater insulin responses to glucose than controls. Glp1r knockdown in β-cells caused glucose intolerance in diet-induced obese mice compared with obese controls, but VSG improved glycemic profiles to similar levels during oral and intraperitoneal glucose challenges in Glp1rβ-cell-ko and Glp1rWT mice. Therefore, even though the β-cell GLP-1R seems to be important for maintaining glucose tolerance in obese mice, in these experiments it is dispensable for the improvement in glucose tolerance after VSG. Moreover, the metabolic physiology activated by VSG can overcome the deficits in glucose regulation caused by lack of β-cell GLP-1 signaling in obesity.

Streptomyces lincolnensis is generally utilized for the production of lincomycin A (Lin-A), a clinically useful antibiotic to treat Gram-positive bacterial infections. Three methylation steps, catalyzed by three different S-adenosylmethionine (SAM)-dependent methyltransferases, are required in the biosynthesis of Lin-A, and thus highlight the significance of methyl group supply in lincomycin production. In this study, we demonstrate that externally supplemented SAM cannot be taken in by cells and therefore does not enhance Lin-A production. Furthermore, bioinformatics and in vitro enzymatic assays revealed there exist two SAM synthetase homologs, MetK1 (SLCG_1651) and MetK2 (SLCG_3830) in S. lincolnensis that could convert l-methionine into SAM in the presence of ATP. Even though we attempted to inactivate metK1 and metK2, only metK2 was deleted in S. lincolnensis LCGL, named as ΔmetK2. Following a reduction of the intracellular SAM concentration, ΔmetK2 mutant exhibited a significant decrease of Lin-A in comparison to its parental strain. Individual overexpression of metK1 or metK2 in S. lincolnensis LCGL either elevated the amount of intracellular SAM, concomitant with 15% and 22% increase in Lin-A production, respectively. qRT-PCR assays showed that overexpression of either metK1 or metK2 increased the transcription of lincomycin biosynthetic genes lmbA and lmbR, and regulatory gene lmbU, indicating SAM may also function as a transcriptional activator. When metK1 and metK2 were co-expressed, Lin-A production was increased by 27% in LCGL, while by 17% in a high-yield strain LA219X.

Background and aim In most species, including humans, food preference is primarily controlled by nutrient value. However, the gut-brain pathways involved in preference learning remain elusive. The aim of the present study, performed in C57BL6/J mice, was to characterize the roles in nutrient preference of two critical elements of gut-brain pathways, i.e. the duodenum and vagal gut innervation. Methods Adult wild-type C57BL6/J mice from a normal-weight cohort sustained one of the following three procedures: duodenal-jejunal bypass intestinal rerouting (DJB), total subdiaphragmatic vagotomy (SDV), or sham surgery. Mice were assessed in short-term two-bottle preference tests before and after 24 h s exposures to solutions containing one of glutamate, lipids, sodium, or glucose. Results DJB and SDV interfered in preference formation in a nutrient-specific manner: whereas normal preference learning for lipids and glutamate was disrupted by both DJB and SDV, these interventions did not alter the formation of preferences for glucose. Interestingly, sodium preferences were abrogated by DJB but not by SDV. Conclusions Different macronutrients make use of distinct gut-brain pathways to influence food preferences, thereby mirroring nutrient-specific processes of food digestion. Specifically, whereas both vagal innervation and duodenal sensing appear critical for generating responses to fats and protein, glucose preferences recruit post-duodenal, vagal-independent pathways in pair with the control of glucose homeostasis. Overall, our data suggest that the physiological processes involved in digesting and absorbing fats, amino acids, and glucose overlap with those mediating learned preferences for each of these nutrients.

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb) is still a leading cause of death worldwide. M. tuberculosis has evolved multipronged strategies to subvert host immune defenses and establish an immunologically privileged niche in macrophages. Rv0426c has been predicted to be an effector involved in the Mtb-host interactions. To investigate the potential role played by Rv0426c, we constructed recombinant M. smegmatis strains with heterologous expression of Rv0426c. We observed that Rv0426c recombinants became more susceptible to various stresses by increasing cell wall permeability, however with elevated early survival rate within macrophages. This was accompanied by decreased levels of pro-inflammatory cytokines and host cell apoptosis. The data suggested that Rv0426c was a new player involved in the interactions between Mtb and macrophages.

The P450 side-chain cleavage enzymes P450scc (Cyp11a) and 11β-hydroxylase (Cyp11b) play important roles in sex steroid and cortisol production. Here, two duplicates of cyp11 genes were identified in Japanese flounder (Paralichthys olivaceus): Pocyp11a and Pocyp11b, respectively. Phylogenetic analysis and amino acid sequence alignment revealed that Pocyp11a and Pocyp11b shared significant identity with sequences of other teleost fish species. The quantitative real-time polymerase chain reaction (qRT-PCR) results indicated that among the studied tissues, brain tissue showed the highest expression of Pocyp11a, followed by kidney and testis tissues, whereas Pocyp11b expression was highest in the testis. The expression patterns of these two genes showed sexual dimorphism, with both genes showing higher expression in the testis than in the ovary. In-situ hybridization analysis demonstrated that Pocyp11a and Pocyp11b mRNA were both detected in oocytes, spermatocytes, and Sertoli cells, indicating that they might be involved in hormone synthesis. The expression levels of Pocyp11a and Pocyp11b were significantly downregulated by treatment with 17α-methyltestosterone (17α-MT) in the testis and ovary in both in vivo and studies. In vivo studies showed that Pocyp11a and Pocyp11b transcripts were suppressed by 17β-estradiol (E2 ) treatment in both the testis and ovary. In addition, in vitro studies showed that the expression level of Pocyp11b was decreased by treatment with E2 , whereas that of Pocyp11a was largely unaffected. Moreover, the expression levels of Pocyp11a and Pocyp11b in the testis cell line were significantly upregulated after NR0b1 and NR5a2 (p < .05) treatment. These results indicate that Pocyp11a and Pocyp11b might play important roles in sex hormone biosynthesis. Our research can assist future studies of the mechanisms of steroid biosynthesis and functional differences between cyp11a and cyp11b in Japanese flounder.

Ghrelin is a 28-peptide hormone that was originally discovered as a ligand for the growth hormone secretagogue receptor (GHSR) and noted for its roles in food intake and feeding behavior. Beyond this, ghrelin may also regulate glucose tolerance and insulin secretion. In times of starvation, ghrelin defends against hypoglycemia. Contrastingly, ghrelin removal increases insulin secretion and improves glucose tolerance in metabolically-stressed mice. Though ghrelin is predominantly produced by stomach X/A cells, it is also made in the pancreas. Within the pancreatic islet, ghrelin is produced by ε-cells and GHSR is predominantly expressed in somatostatin-secreting δ-cells. This positioning suggests that, locally, islet ghrelin could reach interstitial levels that are higher than detected in circulation and act in a paracrine manner to inhibit insulin secretion. Additionally, GHSR has high constitutive activity and may act in the absence of ghrelin. We hypothesized that a ghrelin-GHSR paracrine axis within the islet modulates insulin secretion and tested this using parallel in vivo and ex vivo approaches in genetic rodent models lacking ghrelin (Ghrl-/-) or GHSR (GHSR-/-). Glycemic responses in GHSR-/- and GHSR+/+male mice did not differ after i.p. glucose tolerance tests (IPGTT) after a 6- or 16-hr fast, oral mixed meal tolerance test (MMTT), or insulin sensitivity during insulin tolerance test (ITT). Female GHSR-/- mice had impaired glucose tolerance after 6-hr fast IPGTT compared to GHSR+/+ (glucose AUC, p=0.01) but did not differ from controls during 16-hr fasted IPGTT, MMTT, or ITT. These data suggest constitutive GHSR activity does not mediate in vivo glucose tolerance. Ghrl-/- and Ghrl+/+ male mice did not differ in glycemia in response to IPGTTs, MMTT, or ITT. Female Ghrl-/- mice showed a slight improvement compared to Ghrl+/+ during MMTT (glucose AUC, p=0.04) but did not differ in other tests. Thus, endogenous ghrelin does not substantially alter glucose tolerance. Given that many factors can affect in vivo glucose tolerance, we perifused isolated islets. Here, exogenous ghrelin robustly decreased glucose-stimulated insulin secretion (GSIS, p

Bariatric surgeries including vertical sleeve gastrectomy (VSG) ameliorate obesity and diabetes. Weight loss and accompanying increases to insulin sensitivity contribute to improved glycemia after surgery; however, studies in humans also suggest weight-independent actions of bariatric procedures to lower blood glucose, possibly by improving insulin secretion. To evaluate this hypothesis, we compared VSG-operated mice with pair-fed, sham-surgical controls (PF-Sham) 2 weeks after surgery. This paradigm yielded similar postoperative body weight and insulin sensitivity between VSG and calorically restricted PF-Sham animals. However, VSG improved glucose tolerance and markedly enhanced insulin secretion during oral nutrient and i.p. glucose challenges compared with controls. Islets from VSG mice displayed a unique transcriptional signature enriched for genes involved in Ca(2+) signaling and insulin secretion pathways. This finding suggests that bariatric surgery leads to intrinsic changes within the islet that alter function. Indeed, islets isolated from VSG mice had increased glucose-stimulated insulin secretion and a left-shifted glucose sensitivity curve compared with islets from PF-Sham mice. Isolated islets from VSG animals showed corresponding increases in the pulse duration of glucose-stimulated Ca(2+) oscillations. Together, these findings demonstrate a weight-independent improvement in glycemic control following VSG, which is, in part, driven by improved insulin secretion and associated with substantial changes in islet gene expression. These results support a model in which β cells play a key role in the adaptation to bariatric surgery and the improved glucose tolerance that is typical of these procedures.

Perturbations in carbohydrate, lipid, and protein metabolism contribute to obesity-induced type 2 diabetes (T2D), though whether alterations in ketone body metabolism influence T2D pathology is unknown. We report here that activity of the rate-limiting enzyme for ketone body oxidation, succinyl-CoA:3-ketoacid-CoA transferase (SCOT/Oxct1), is increased in muscles of obese mice. We also found that the diphenylbutylpiperidine pimozide, which is approved to suppress tics in individuals with Tourette syndrome, is a SCOT antagonist. Pimozide treatment reversed obesity-induced hyperglycemia in mice, which was phenocopied in mice with muscle-specific Oxct1/SCOT deficiency. These actions were dependent on pyruvate dehydrogenase (PDH/Pdha1) activity, the rate-limiting enzyme of glucose oxidation, as pimozide failed to alleviate hyperglycemia in obese mice with a muscle-specific Pdha1/PDH deficiency. This work defines a fundamental contribution of enhanced ketone body oxidation to the pathology of obesity-induced T2D, while suggesting pharmacological SCOT inhibition as a new class of anti-diabetes therapy.

The current study aimed to explore the functions and roles of microRNA-193b (miR-193b) in the myocardium with ischemia-reperfusion (I/R) injury and a potential therapeutic method for myocardial I/R injury. The mice were subjected to myocardial I/R with or without miR-193b pretreatment. The infarct size and myocardial enzymes were detected. The terminal deoxynucleotidyl transferase dUTP nick-end labeling assay was conducted to investigate the effect of miR-193b on cardiomyocyte apoptosis. The expression levels of miR-193b and mastermind-like 1 (MAML1) were validated by quantitative real-time polymerase chain reaction and Western blot analysis. The results suggested that the miR-193b expression level was significantly downregulated in the myocardium with I/R injury compared with control group. miR-193b overexpression is able to reduce infarct size and myocardial enzymes after myocardial I/R injury. Furthermore, overexpression of miR-193b could alleviate the apoptosis level after myocardial I/R injury. Taken together, the present study demonstrated that upregulated miRNA-193b alleviated myocardial I/R injury via targeting MAML1.

The ghrelin-producing ε cell represents the fifth endocrine cell type in human pancreatic islets. The abundance of ε cells in adult pancreas is extremely low, which has hampered the investigation on the molecular pathways regulating the development and the function of this cell type. In this study, we explored the molecular features defining the function of pancreatic ε cells isolated from adult nondiabetic donors using single-cell RNA sequencing technology. We focus on transcription factors, cell surface receptors, and genes involved in metabolic pathways that contribute to regulation of cellular function. Furthermore, the genes that separate ε cells from the other islet endocrine cell types are presented. This study expands prior knowledge about the genes important for ε cell functioning during development and provides a resource to interrogate the transcriptome of this rare human islet cell type.

Persisters, stochastic dormant variants of normal bacteria cell, represent a significant portion of the survivors upon exposure to antibiotics and other environmental stresses, which contributes substantially to high level antibiotics tolerance. Glutamine is a crucial component of the Mycobacteria nitrogen pool that is indispensable for survival upon stresses. To study whether a synergistic effect exists between glutamine and antibiotics against Mycobacterial persisters, the efficacy of rifampicin alone or together with exogenous glutamine upon Mycobacterium smegmatis mc2 155 persisters was monitored. The result showed that glutamine decreases M. smegmatis tolerance to rifampicin upon starvation. The reactive oxygen species level of the strains treated with rifampicin and glutamine increased. The synergism of glutamine and rifampicin to kill persisters might derive from altering the oxidative phosphorylation and TCA cycle, as both evidenced by both ATP level increase and transcriptome change. Glutamine might represent a synergistic agent of rifampicin to kill Mycobacteria persisters.

High dietary iron intake is associated with impaired islet-cell function and increased risk of type 2 diabetes mellitus (T2DM) while iron chelation leads to improved glucose tolerance. Bariatric surgery has multiple metabolic benefits but iron-deficiency anemia is a common occurrence following surgery, presumably because of preferential iron absorption in the upper intestine. We hypothesized that a duodenal-jejunal bypass (DJB) procedure that bypasses the proximal small bowel and vertical sleeve gastrectomy (VSG) that eliminate 80% of the stomach without altering the intestine would have differential effects on iron metabolism and glucose homeostasis. C57BL6J mice were fed either high- (2000 ppm) or low-iron diet (4 ppm) for 16 weeks before DJB (n=6 per diet group), VSG (n=6/7 per diet group) or sham surgery (n = 11 per diet group). Intraperitoneal glucose, and insulin, tolerance tests were performed at Day 0, 10 and 20 after surgery. Compared to animals given a low-iron diet, 4 months of high-iron diet significantly impaired glucose tolerance (p Disclosure J. Niu: None. J.D. Douros: None. J. Campbell: None. M. Capozzi: None. S.M. Gray: None. F. Lorenzo: None. D. D'Alessio: Consultant; Self; Intarcia Therapeutics, Inc., Eli Lilly and Company, Merck & Co., Inc., Novo Nordisk A/S. D. McClain: None. J. Tong: None.

In the online published article, row value “pIB139-metK1-metK2” in table 1 has been processed incorrectly. The correct table is given below

Reductions in calorie intake contribute significantly to the positive outcome of bariatric surgeries. However, the physiological mechanisms linking the rerouting of the gastrointestinal tract to reductions in sugar cravings remain uncertain. We show that a duodenal-jejunal bypass (DJB) intervention inhibits maladaptive sweet appetite by acting on dopamine-responsive striatal circuitries. DJB disrupted the ability of recurrent sugar exposure to promote sweet appetite in sated animals, thereby revealing a link between recurrent duodenal sugar influx and maladaptive sweet intake. Unlike ingestion of a low-calorie sweetener, ingestion of sugar was associated with significant dopamine effluxes in the dorsal striatum, with glucose infusions into the duodenum inducing greater striatal dopamine release than equivalent jejunal infusions. Consistently, optogenetic activation of dopamine-excitable cells of the dorsal striatum was sufficient to restore maladaptive sweet appetite in sated DJB mice. Our findings point to a causal link between striatal dopamine signaling and the outcomes of bariatric interventions.

R-spondin 1 (Rspo1) is a potential female-determining gene in mammals that could regulate the Wnt/β-catenin signaling pathway. The deletion of Rspo1 causes sex reversal in females. To investigate sexual determination and differentiation, we cloned and analyzed the Rspo1 gene in Cynoglossus semilaevis. Phylogenetic and gene structure analyses revealed that Rspo1 gene exhibited high sequence conservation and contained an N-terminal signal peptide, two furin-like cysteine-rich domains (FU1 and FU2), a thrombospondin type 1 repeat, and a C-terminal region enriched with basic charged amino acids. qRT-PCR revealed that Rspo1 expressed sexual dimorphism in gonad, with higher expression levels in the ovary than in the testis, thus, suggesting the involvement of Rspo1 in gonad differentiation. In situ hybridization results demonstrated that Rspo1 was expressed in premature germ cells, including spermatogonia and spermatocytes in the testis and stage II and stage III oocytes in the ovary. The methylation levels in two CpG sites of Rspo1 promoter significantly differed among females, males, and pseudomales. After 30days of exposure to high temperature, the expression of Rspo1 significantly decreased in female individuals, some of which were prone to males. However, no difference of Rspo1 gene expression was observed between the control group and high-temperature group in males. These preliminary findings suggested that Rspo1 played a crucial role in sex determination and development. This study laid the groundwork for further sex control breeding techniques in C. semilaevis.

Objective In Chinese adults, the trend of central obesity and its longitudinal association with hypertension, independent of general obesity, was examined. Methods A 12-year longitudinal analysis was conducted using data retrieved from the China Health and Nutrition Survey. This study examined 6096 individuals (normotensive in 1997) who were followed up with in 2000, 2004, 2006, and 2009. Prevalence of hypertension in 2009 was predicted by baseline central obesity and waist circumference changes during a 12-year follow-up period along with confounding covariates using multiple logistic regressions. Results Between 1997 and 2009, the prevalence of central obesity increased from 17.3% to 39.4% and was highest among individuals ≥ 60 years of age in 1997. By 2009, 26.8% of the participants developed hypertension. The odds ratio of developing hypertension during the 12-year study period for Chinese adults with central obesity at baseline was 1.79 (95% confidence interval = 1.36–2.35) compared to those without central obesity, controlling for general obesity, demographics, smoking/drinking behavior, and fat intake. Conclusions Among Chinese adults, central obesity increases the risk for developing hypertension later in life, even after controlling for general obesity, smoking, drinking, and high fat intake among other factors. Waist circumference should be targeted in the efforts of hypertension prevention.

Little is known on the extent of weight misperception and its relation with dietary intake among Chinese youth.The study aimed to investigate extent and correlates of weight misperception and its relation with dietary intake among Chinese youth.Data pertaining to Chinese youth, 6-17 years of age, from the 2004-2009 China Health and Nutrition Survey (N = 3,923) were analyzed using STATA version 12.1. The heights and weights of the participants were measured by well-trained health workers at the participants' homes or a local clinic following the reference protocol recommended by the World Health Organization. The dietary intake data of the participants were collected on three consecutive days at both the household and individual levels.Of the children 6-11 years of age, 18.9% were underweight and 15.3% were overweight. Among the children 12-17 years of age, 18.3% were underweight and 8.1% were overweight. Less than 60 % of Chinese youth accurately estimated their weight status. Nutrition knowledge was positively related to a perception of being overweight (adjusted odds ratio [AOR] = 1.98, p = .007) among children 12-17 years of age. A perception of being overweight was positively associated with fat and protein intake among children 6-11 years of age (p .05).A discrepancy exists between the actual and perceived weight status of Chinese youth. Efforts are needed in China to promote accurate weight perception, healthy weight, and eating behaviors.

There is a paucity of recent data on Chinese childhood overweight and underweight prevalence especially since 2004.The purpose of this study was to examine trends in underweight and overweight/obesity ("overweight" hereafter) prevalence and energy balance-related behaviors of Chinese youth from 2004 to 2009.Data from the China Health and Nutrition Survey, 2004-2009 (N = 4,061 students aged 6-18 years), were analyzed. Trained health workers took anthropometric measures at the participant's house or at a local clinic following a reference protocol recommended by the World Health Organization. The international age- and sex-specific body mass index reference standard proposed by the International Obesity Task Force was used to define underweight and overweight children in this study.Among 6- to 11-year-old boys, underweight prevalence increased from 14.5% (2004) to 20.1% (2009, p = 0.068). Among 12- to 18-year-old boys, however, overweight prevalence increased from 7.5 to 12.6% (p = 0.034). From 2004 to 2009, after-school sedentary behavior increased from 2.3 to 3.4 h/day for 6- to 11-year-olds (p 0.001) and from 2.2 to 3.1 h/day for 12- to 18-year-olds (p 0.01). Meanwhile, the total energy intake decreased 7% for 6- to 11-year-olds (p 0.05) and 10% for 12- to 18-year-olds (p 0.01).Both underweight and overweight Chinese students are increasing, with underweight increases more pronounced in 6- to 11-year-olds and overweight increases more pronounced in 12- to 18-year-olds. Nationwide efficacious interventions are needed that improve the diet, decrease sedentary behavior, and encourage a healthy and realistic body image in Chinese youth.

The nodal signaling pathway has been shown to play crucial roles in inducing and patterning the mesoderm and endoderm, as well as in regulating neurogenesis and left-right axis asymmetry. Here, we present the first complete cDNA and genomic sequences as well as the promoter predication of the Dnah9 gene in the Japanese flounder. The 15,558-bp-long cDNA is divided into 96 exons and spread over 138 kb of genomic DNA. Protein sequence comparison showed that it shares higher identity with other vertebrate orthologs, with an ATP binding dynein motor, AAA domain and microtubule binding stalk of dynein motor. Dnah9 exhibited maternal and ubiquitous expression in all cells of the early development stages, but became concentrated in the head at 1 DAH, as identified by qRT-PCR and in situ hybridization methods. Furthermore, after nodal signaling was inhibited, the level of Southpaw did not change significantly at early development stage (50 % epiboly) but increased significantly at late stages (27-somite stages and 1 DAH), as well as the expression of Lefty, an inhibitor of nodal signaling, increased continuously. On the other hand, the expression level of Dnah9 decreased. The transcription factor binding site of FAST-1 (SMAD interacting protein) was identified in the transcription region of Dnah9 by the promoter analysis, which might format the complexes of SMADs, FAST-1 and the transcription region of Dnah9 served as a bridge of Dnah9 and nodal signaling. All evidences indicated that Dnah9 might be downstream of nodal during the early development stages, and an indirect function through SMADs for nodal signaling pathway.

Antarcticite, CaCl2 · 6H2O, is an ideal phase change material (PCM) due to its high-energy storage density and good thermal conductivity. In this chapter, supercooling and subsequent solidification behavior of antarcticite are studied based on the cooling curve method and DSC measurement. The results show that the minor SrCl2 · 6H2O as the nucleator and carboxymethyl cellulose as the thickening agent could significantly reduce supercooling and partly restrain the phase separation. Moreover, we incorporated antarcticite as PCM into building envelopes in four different cases, and the simulation of the heat transfer processes showed that the temperature fluctuation could be reduced to about 2 °C in the best case.

Background: Internet-based interventions are more cost-effective than conventional interventions and can provide immediate, easy-to-access, and individually tailored support for behavior change. Waist circumference is a strong predictor of an increased risk for a host of diseases, such as hypertension, diabetes, and dyslipidemia, independent of body mass index. To date, no study has examined the effect of Internet-based lifestyle interventions on waist circumference change. Objective: This study aimed to systematically review the effect of Internet-based interventions on waist circumference change among adults. Methods: This meta-analysis reviewed randomized controlled trials (N=31 trials and 8442 participants) that used the Internet as a main intervention approach and reported changes in waist circumference. Results: Internet-based interventions showed a significant reduction in waist circumference (mean change –2.99 cm, 95% CI −3.68 to −2.30, I 2 =93.3%) and significantly better effects on waist circumference loss (mean loss 2.38 cm, 95% CI 1.61-3.25, I 2 =97.2%) than minimal interventions such as information-only groups. Meta-regression results showed that baseline waist circumference, gender, and the presence of social support in the intervention were significantly associated with waist circumference reduction. Conclusions: Internet-based interventions have a significant and promising effect on waist circumference change. Incorporating social support into an Internet-based intervention appears to be useful in reducing waist circumference. Considerable heterogeneity exists among the effects of Internet-based interventions. The design of an intervention may have a significant impact on the effectiveness of the intervention. [J Med Internet Res 2015;17(7):e181]