Enterobacter sp. strain EA-1 is an electrochemically active bacterium isolated from tropical sediment in Singapore. Here, the annotated draft genome assembly of the bacterium is reported. Whole-genome comparison indicates that Enterobacter sp. EA-1, along with a previously sequenced Enterobacter isolate from East Asia, forms a distinct clade within the Enterobacter genus. Enterobacter belong to the class Gammaproteobacteria and are rod-shaped, Gram-negative, motile, facultative anaerobes. They are commonly found in soil, water, sewage, and the intestines of animals and humans and have been associated with nosocomial infections  broth at room temperature and with shaking at 150 rpm. The Wizard genomic DNA purification kit  was used according to the protocol for Gram-negative bacteria supplied by the manufacturer.The draft genome sequence was determined by shotgun sequencing on a MinION Mk1B sequencer  using R9 chemistry and library protocol SQK-LSK108. Raw sequencing data were assembled using Canu version 1.3 , manuallEnterobacter cloacae ATCC 13047, Enterobacter cloacae SBP-8, and Enterobacter sp. strain FY-07. However, a whole-genome comparison indicated an average nucleotide identity (E. cloacae ATCC 13047 (NCBI accession number CP001918), 78.09% compared to E. cloacae SBP-8 (NCBI accession number CP016906), and 93.38% compared to FY-07 (NCBI accession number CP012487), suggesting that EA-1 forms, together with FY-07, a distinct clade within the Enterobacter genus related to but separate from E. cloacae.The draft genome of EA-1 is composed of 5,145,523 bp, with an average G+C content of 54.56%, and it contains 81 tRNAs, 22 rRNAs organized in 7 operons, and 1 transfer-messenger RNA (tmRNA). A comparison of the 16S rRNA gene sequence revealed >98% identity to the 16S rRNA genes of http://www.ebi.ac.uk/Tools/psa/) using the EMBOSS Needle tool . In FY-07, these genes are arranged in three BC-producing operons: bcsI, bcsII, and bcsIII , AKI40_0197 (CWS02_01215), AKI40_0198 (CWS02_01220), AKI40_0199 (CWS02_01225), AKI40_0200 (CWS02_01230), AKI40_0201 (CWS02_01235), and AKI40_0202 (CWS02_01240). For bcsII, the FY-07 and corresponding EA-1 locus tags (in parentheses) are AKI40_0206 (CWS02_01260), AKI40_0207 (CWS02_01265), AKI40_0208 , AKI40_0209 (CWS02_01285), AKI40_0210 (CWS02_01290), and AKI40_0211 (CWS02_01295). In the case of the EA-1 locus tag corresponding to FY-07’s AKI40_0208, multiple ORFs were stitched together during nucleotide alignment in order to identify the complete homologous gene sequence which was interrupted due to frameshifting errors associated with the sequencing technology used. For bcsIII, the FY-07 and corresponding EA-1 locus tags (in parentheses) are AKI40_0894 (CWS02_05060), AKI40_0893 (CWS02_05055), AKI40_0892 (CWS02_05050), and AKI40_0891 (CWS02_05045). For other BC-related genes (not operon-associated), the FY-07 and corresponding EA-1 locus tags are AKI40_0203 (CWS02_01245), AKI40_0204 (CWS02_01250), and AKI40_0205 (CWS02_01255).In-detail genome alignments between EA-1 and FY-07 using the Artemis Comparison Tool , are alsd bcsIII . In bcsICP025776.The whole-genome shotgun project was deposited in NCBI under the accession number


Salmonella enterica subsp. diarizonae serovar 61:k:1,5,(7) strain CRJJGF_00165 , isolated from ground beef in 2007.Here, we report a 4.78-Mb draft genome sequence of the  Salmonella enterica subsp. diarizonae serovar 61:k:1,5,(7) strain CRJJGF_00165  distinguishes it from the phylogenetically distinct monophasic S. enterica subsp. IIIa serovar 61:k:1,5,(7)  is mostly associated with reptiles  strain CRJJGF_00165 was isolated from ground beef using standard microbiology techniques. SMART-PCR was used to serotype this isolate  and tRNAs were predicted with Prodigal . DNA libraries were constructed using the Nextera XT DNA preparation kit, and paired-end sequencing was performed on the Illumina HiSeq2500 platform  using a 500-cycle MiSeq reagent kit. A total of 3,848,000 reads were generated. Reads were g Velvet  into 108Prodigal  and ARAGProdigal , respectProdigal , CRISPRFProdigal , and PHAProdigal , respectRG-ANNOT . The genS. enterica subsp. IIIb serovar 61:k:1,5,(7) strain CRJJGF_00165 has been deposited in GenBank (NCBI) under the accession number JQYQ00000000. The version described here is the first version.The genome sequence of


Addendum to: Translational Psychiatry (2017) 7, e1001;doi:10.1038/tp.2016.263; published online 17 January 2017post hoc tests for clarifying the direction of the three MANOVA results thatfound significant associations between functional connectivity changes and long-termclinical outcomes. In each of the three regression tests, greater increases infunctional connectivity were associated with improved long-term clinical outcomes, asascertained by positive beta weights for the following regressors: longitudinal positivepsychotic symptoms =2.79;P=0.015), longitudinal affective symptoms =2.36; P=0.035) and subjective long-termrecovery =2.56;P=0.024). These tests replace the correlation tests originally reportedin the article.After publication, the authors determined that regression tests are the appropriate


In sub-Saharan Africa, 20%–25% of people starting antiretroviral therapy (ART) have severe immunosuppression; approximately 10% die within 3 months. In the Reduction of EArly mortaLITY (REALITY) randomized trial, a broad enhanced anti-infection prophylaxis bundle reduced mortality vs cotrimoxazole. We investigate the contribution and timing of different causes of mortality/morbidity.Participants started ART with a CD4 count <100 cells/µL; enhanced prophylaxis comprised cotrimoxazole plus 12 weeks of isoniazid + fluconazole, single-dose albendazole, and 5 days of azithromycin. A blinded committee adjudicated events and causes of death as  tuberculosis, cryptococcosis, severe bacterial infection (SBI), other potentially azithromycin-responsive infections, other events, and unknown.P < .05) but not tuberculosis, SBI, potentially azithromycin-responsive infections, or other causes (P > .3); and reduced nonfatal/fatal tuberculosis and cryptococcosis (P < .05), but not SBI, other potentially azithromycin-responsive infections, or other events (P > .2).Median pre-ART CD4 count was 37 cells/µL. Among 1805 participants, 225 (12.7%) died by week 48. Fatal/nonfatal events occurred early (median 4 weeks); rates then declined exponentially. One hundred fifty-four deaths had single and 71 had multiple causes, including tuberculosis in 4.5% participants, cryptococcosis in 1.1%, SBI in 1.9%, other potentially azithromycin-responsive infections in 1.3%, other events in 3.6%, and unknown in 5.0%. Enhanced prophylaxis reduced deaths from cryptococcosis and unknown causes (Enhanced prophylaxis reduced mortality from cryptococcosis and unknown causes and nonfatal tuberculosis and cryptococcosis. High early incidence of fatal/nonfatal events highlights the need for starting enhanced-prophylaxis with ART in advanced disease.ISRCTN43622374. In sub-Saharan Africa, 20%–25% of human immunodeficiency virus (HIV)–infected individuals starting antiretroviral therapy (ART) have a CD4 count <100 cells/µL . In the The REALITY trial ISRCTN43622374) recruited 1805 HIV-infected ART-naive adults and children ≥5 years (98% aged ≥13 years) from Zimbabwe, Uganda, Malawi, and Kenya with a CD4 count <100 cells/µL. In brief, participants were randomized 1:1 to enhanced prophylaxis vs standard prophylaxis (cotrimoxazole) ), other events (n = 63 [3.6%]), SBI (n = 33 [1.9%]), other potentially azithromycin-responsive infections (n = 23 [1.3%]), and cryptococcosis (n = 20 [1.1%]). Sixty-seven (29.8%) deaths were adjudicated as compatible with IRIS. Enhanced prophylaxis reduced all-cause mortality by 24 weeks by 27%  log-rank 8.4% wereP = .06) . There w4 and 48 .P = .007), cryptococcosis , and IRIS-compatible events  (P = .26), potentially azithromycin-responsive infections (P = .34), or other events (P = .81). Although occurring at much higher rates  events of each category occurred in 674 (37.3%) participants. The most common were other events (n = 479 participants [27.0%]) , followe = .001) , but noter rates , similarer rates . Rates fAt 48 weeks post–ART initiation, rates of fatal/nonfatal events were lowest  and participants with lower baseline CD4 count (P = .02) or self-reported vomiting (P = .005) (P = .03); those with lower baseline CD4 count (P = .05), creatinine clearance (P < .001), or albumin (P = .01); higher bilirubin (P = .05); without previous healthcare contact (P = .04); or with wasting/severe weight loss (P = .02), participant-reported fever (P = .08), or problems with mobility (P = .05) or self-care (P = .05). There was no effect of water source or household toilet type on either cause of death (P > .3).Given the impact of enhanced prophylaxis on deaths from cryptococcosis and unknown causes, we considered whether predictors of these 2 categories of death were similar, suggesting unascertained cryptococcal disease could be driving these effects. Deaths from cryptococcosis were more common in standard prophylaxis ( = .005) . Deaths Considering the 1716 participants alive and in follow-up at week 4, median last postbaseline VL before death was 95  copies/mL , with reIn the REALITY trial of patients starting ART with severe immunosuppression in sub-Saharan Africa, morbidity/mortality rates were high in the first month after starting ART, regardless of type of event, and dropped exponentially thereafter. However, even within a randomized trial with intensive follow-up, structured clinical narratives, and independent endpoint review, causes of death were difficult to ascertain and the commonest category was unknown. Furthermore, multiple causes were identified in almost one-third of deaths, highlighting the complexity of clinical presentations in patients starting ART with advanced immunosuppression.REALITY showed that enhanced prophylaxis reduces mortality by 27% , due to Of note, 62.7% of participants had no serious morbidity/mortality during their first 48 weeks on ART, despite very low pre-ART CD4 and high VL. Nevertheless, their low CD4 put them at substantially increased risk of impaired short- and long-term immunological response, reinforcing the importance of undertaking baseline CD4 to identify these patients. Where CD4 is becoming less available, our data reinforce the need to further develop CD4 point-of-care tests which at least qualitatively identify the most immunosuppressed individuals. Alternatively, total lymphocyte counts could be used to identify at least some of those at greatest risk , 21.Given current WHO guidance focusing on diagnostic tools to rule out opportunistic infections before ART initiation , our praClinical Infectious Diseases online. Consisting of data provided by the authors to benefit the reader, the posted materials are not copyedited and are the sole responsibility of the authors, so questions or comments should be addressed to the corresponding author.Supplementary materials are available at Supplementary Material 1Click here for additional data file.


Juniperus phoenicea (Arar), Anastatica hierochuntica (Kaff Maryam), and Citrullus colocynthis  with a set of reference compounds with established modes of action. Cluster analyses of the cytological profiles of the tested compounds suggested that these plants contain possible topoisomerase inhibitors that could be effective in cancer treatment. Using histone H2AX phosphorylation as a marker for DNA damage, we discovered that some of the compounds induced double-strand DNA breaks. Furthermore, chemical analysis of the active fraction isolated from Juniperus phoenicea revealed possible anti-cancer compounds. Our results demonstrate the usefulness of cell-based phenotypic screening of natural products to reveal their biological activities.Natural products have been used for medical applications since ancient times. Commonly, natural products are structurally complex chemical compounds that efficiently interact with their biological targets, making them useful drug candidates in cancer therapy. Here, we used cell-based phenotypic profiling and image-based high-content screening to study the mode of action and potential cellular targets of plants historically used in Saudi Arabia’s traditional medicine. We compared the cytological profiles of fractions taken from  Cancer is a leading cause of morbidity and mortality worldwide, exceeding the number of cases of illness and death due to HIV/AIDS, malaria, and tuberculosis . In 2012High-content screening (HCS) optimizes the discovery of biologically active small molecules and their subsequent development into therapeutic compounds by the use of automated microscopy in conjunction with image analysis. A high-throughput platform based on image-processing software that reviews images from automated fluorescence microscopy –10, HCS Anastatica hierochuntica (Kaff Maryam), Juniperus phoenicea (Arar), and Citrullus colocynthis , that have been used in traditional medicine in Saudi Arabia. Previous reports reported anti-cancer activity in Juniperus phoenicea and Citrullus colocynthis .–31.1H NM 4 and 5 , 33. TheThis study demonstrates the capability of HCS for cytological profiling for drug discovery in natural products. In recent years, HCS has developed from a promising concept into an efficient methodology and indispensable tool. HCS now can be implemented during the early drug discovery process as a result of its recent technological advances , 34.C. colocynthis, J. phoenicea, and A.hierochuntica. We used a library of known small molecules with assigned modes of action as reference compounds. We compared their cytological profiles with profiles retrieved from the fractionated plant extracts. We were able to predict the mode of action of pure compounds as well as fractionated extracts -βDgalactopyranoside triacetate and Estra-1,3,5(10)-triene-3,6beta,17beta-triol triacetate. Estra-1,3,5(10)-triene-3,6beta,17beta-triol triacetate (PubChem CID: 5756) is used as anti-inflammatory drug but no anti-inflammatory activity has been reported for Methyl6-O--β-Dgalactopyranoside triacetate compound.The 2,2-dimethoxybutane (PubChem CID: 137941) found in the JUN_C2_60% fraction was found to be toxic to microbial membranes –61, but polymers . This mae needed . 3-Trifl effects , 67. It  effects , in agre Tunisia . Anti-in Tunisia . Ethyl 4 Tunisia . Of the  Tunisia –75. SphiOur results demonstrate the power of HCS for drug discovery. HCS allows traditionally used medicinal plants to be assessed for the presence of bioactive compounds. Cytological profiles allow us to identify topoisomerase II inhibitor activity in the plant fractions that we tested to verify the presence of anti-cancer compounds in the plants. Further studies are needed to evaluate and better characterize the active compounds identified in our study. This can be done by guided fractionation using HPLC to isolate and eliminate the active from the non-active compounds and to retest the active compounds with HCS. Furthermore, active compounds can be synthesized and tested in vivo.S1 Table(DOCX)Click here for additional data file.S2 TableThis figure were adapted from the cited publication.(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 Table(DOCX)Click here for additional data file.S6 Table(DOCX)Click here for additional data file.S1 FigEight peaks were identified from the LC-MS chromatogram: (1) 185.98322 m/z, (2) 144.98225 m/z, (3) 144.98225 m/z, (4) 288.28995 m/z, (5) 256.26364 m/z, (6) 387.18086 m/z, (7) 415.21219 m/z, and (8) 637.30585 m/z.(TIFF)Click here for additional data file.S2 Fig4H5NO4S.Acesulfame-K, Chemical Formula: C(TIFF)Click here for additional data file.S3 Fig2H6ClO3P.Ethephon, Chemical Formula: C(TIFF)Click here for additional data file.S4 FigSphinganine, Chemical Formula: C17H37NO2.(TIFF)Click here for additional data file.S5 Fig16H33NO.Palmitic amide, Chemical Formula: C(TIFF)Click here for additional data file.S6 Fig22H26O6.Burseran or (+)Eudesmin, Chemical Formula: C(TIFF)Click here for additional data file.S7 Fig24H30O6.Estra-1,3,5(10)-triene-3,6beta,17beta-triol triacetate. Chemical Formula: C(TIFF)Click here for additional data file.S8 Fig29H48O15.Methyl 6-O--β-D-galactopyranoside. Chemical Formula: C(TIFF)Click here for additional data file.S9 FigThe spectrum was recorded at room temperature using a 600-MHz NMR spectrometer.(TIFF)Click here for additional data file.S10 FigThe spectrum was recorded at room temperature using a 600-MHz NMR spectrometer. This figure verifies the MS finding of the -6-methyl-β-D-glucopyranuronosyl]-β-D-galactopyranoside) molecules with signals of several CH3 groups around 1 ppm in (TIFF)Click here for additional data file.S11 FigA) Estra-1,3,5(10)-triene-3,6beta,17beta-triol triacetate B) Burseran C) (+)Eudesmin D) Extended aromatic region of the NMR spectrum.(TIFF)Click here for additional data file.S12 Fig(TIFF)Click here for additional data file.S13 Fig(TIFF)Click here for additional data file.S14 Fig(TIFF)Click here for additional data file.


Magnetospirillum sp. 15-1. This strain was isolated from a planted fixed-bed reactor based on its ability to degrade toluene under anaerobic conditions. The genome assembly consists of 5.4 Mb in 28 contigs and 5,095 coding sequences containing the genes involved in anaerobic toluene degradation.Here, we report the draft genome sequence of  Betaproteobacteria strains, represented by Thauera aromatica and Azoarcus sp.  continuously fed with toluene (De novo assembly was conducted with the A5-miseq pipeline (http://www.geneious.com)  (coding percentage 98.82%). The assembled sequences were functionally annotated using the RAST online service  , generat (M. magnetotacticum MS-1 (86%) (Magnetospirillum sp. XM-1 (Magnetospirillum sp. SO-1 (89%)  (M. bellicus VDY (77%) (Magnetospirillum sp. WD (77%)  , M. magn-1 (86%) , Magnetosp. XM-1  (88%), M-1 (89%) , and M. -1 (76%)  than to DY (77%)  and MagnWD (77%) . This fibss, bbs, and bam), as observed for Thauera aromatica K172 (bss operon of strain 15-1 against other anaerobic toluene degraders were performed through BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi). Results showed similarities of 99% to Magnetospirillum sp. TS-6 and 80% to both T. aromatica K172 (Azoarcus sp. (Magnetospirillum strains (excluding strain 15-1) showed that none of these strains contain genes related to aromatic compound degradation pathways.The genes for degradation of toluene under anaerobic conditions were found to be organized in three operons on the chromosome  under the accession no.


Globally, endometrial cancer is the sixth leading cause of female cancer-related deaths. Non-atypical endometrial hyperplasia (EH), has a lifetime progression rate to endometrial cancer ranging from less than 5%, if simple without atypia, to 40%, if complex with atypia. Site specific, long-acting intrauterine devices (IUDs) provide fertility sparing, progestin-based EH medical management. It is unclear which IUD is most beneficial, or if progesterone sensitizing metformin offers improved outcomes. For resolution, PubMed searches for “Mirena” or “Metformin,” “treatment,” “endometrial hyperplasia,” or “stage 1 endometrial cancer,” were performed, yielding 33 articles. Of these, 19 articles were included. The 60 mg high-dose frameless IUD/20 mcg levonorgestrel has achieved sustained regression of Grade 3 endometrial intraepithelial neoplasia for 14 years. Case series on early stage endometrial cancer (EC) treatment with IUDs have 75% or greater regression rates. For simple through complex EH with atypia, the 52 mg-IUD/10–20 mcg-LNG-14t has achieved 100% complete regression in 6-months. Clearly, IUDs have an outcome advantage over oral progestins. However, studies on metformin for EH, and of progestins or metformin for early stage EC management are underpowered, with inadequate dose ranges to achieve significant differences in, or optimal outcomes for, the treatment modalities. Therefore, outcomes from the feMMe trial for the 52 mg-IUD/10–20 mcg-LNG-14t and metformin will fill a gap in the literature. Endometincrease . Therefoincrease . Insulinincrease . Based on unopposed estrogen driving malignant endometrial proliferation, progestins have been the mainstay for fertility sparing medical management of EH and Stage I, grade I EC ,8,12,13.p < 0.04, after adjusting for age, clinical stage, grade, chemotherapy treatment, radiation treatment and presence of hyperlipidemia [Metformin has been successfully used in a few cases of progestin nonresponsive atypical EH . Cell liipidemia . With inipidemia . This waipidemia . Progestipidemia . As metfIt will be shown that oral continuous or intermittent progestin has lower regression rates (higher progression rates), than do progestin containing IUDs. In other words, the locally- long-acting intrauterine progestin delivery provided by the 52 mg-IUD/10–20 mcg-LNG-14t, the 60 mg-IUD/14 mcg-LNG, the 60 mcg-HD-frameless-IUD/20 mcg-LNG and the 60 mcg-LD-frameless-IUD/14 mcg-LNG more effectively medically manages EH than does continuous or intermittent oral progestin. PubMed searches on June 3, 2017, search terms “Mirena treatment endometrial hyperplasia” and “metformin treatment endometrial hyperplasia” with the parameters English language, free full text, published from 2012 onwards yielded 25 articles. Of these, 1 was on contraception, 1 was redundant, 1 was concerned with diagnosis, 1 focused on PCOS, 1 focused on hysterectomy in obese patients, 1 included hysteroscopic resection, 1 explored progestin resistance, and 18 were included. PubMed searches on February 11, 2017, search terms “Mirena treatment stage 1 endometrial cancer” and “metformin treatment stage 1 endometrial cancer” with the parameters English language, free full text, published from 2012 onwards, yielded 8 articles. Of these, 3 were excluded as a duplicate, 2 were concerned with hysteroscopic resection as treatment or pretreatment followed by progestin treatment, 1 was concerned with ovarian cancer and 1 was concerned with advanced EC, leaving 1 inclusion. From the four PubMed searches, 19 articles were included, 14 articles were excluded. Hand search yielded seven additional included articles as shown in Progestins modulate endometrial glands’ secretory differentiation, inhibit estrogen receptor function and endometrial cell mitosis, and are pro-apoptotic . ProgestGrade 2 endometrial endometrioid adenocarcinoma in an 18-year old nullipara with Class III obesity, NIDDM and PCOS was treated with a 52 mg-IUD/10–20 mcg-LNG-14t . CompletA multi-center randomized controlled trial (RCT) with 170 participants compared the 52 mg-IUD/10–20 mcg-LNG-14t to 10 mg MPA for 10 days monthly or continuously for EH treatment . Patholop = 0.001 [p = 0.001 [p = 0.30 [For further surveillance, 153 participants from the above trial participated in a national, multicenter RCT of the 52 mg-IUD/10-20 mcg-LNG-14t compared to cyclic MPA 10 mg for 10 days per cycle, and continuous MPA 10 mg daily for 6-month’s treatment followed by reevaluation at 6, 12, 18, and 24 months . Of the  = 0.001 . Consist = 0.001 . Howeverp = 0.30 . It is ip = 0.30 .p = 0.013, this study, which did not use intent to treat analysis, was underpowered for the 18.9 percentage-point regression rate difference to achieve statistically significance [A RCT of 60 participants in total, compared EH treatment with the 52 mg-IUD/10–20 mcg-LNG-14t to MPA 10 mg orally daily for 12 days monthly for 3-months [p < 0.01) and 55.7% cyclic NET for simple hyperplasia respectively at 12 months [At 1-year follow up, a randomized controlled trial (RCT) with 59 patients assigned to the 52 mg-IUD/10–20 mcg-LNG-14t versus 61 patients assigned to cyclic norethindrone (NET) 15 mg daily for 3-week cycles, indicated that the 52 mg-IUD/10–20 mcg-LNG-14t was more effective than cyclic NET medical management of non-atypical EH ,21. InteA single case series of 20 women with non-atypical and atypical EH, achieving sustained, complete regression for an average of 32-months with the 60mg-IUD/14 mcg-LNG or 60 mg-HD-frameless-IUD/20 mcg-LNG has been reported . A 44-yep < 0.0001 [p < 0.00001), however, this was predominantly spotting which resolved in 1-year [p = 0.08) [p = 0.28), or cancer-induced death,  [Meta-analysis of three RCT with a total of 359 participants found that the 52 mg-IUD/10–20 mcg-LNG-14t prevented tamoxifen-induced endometrial polyp formation, odds ratio (OR) 0.18, 95% confidence interval (CI): 0.13 to 1.02, < 0.0001 . Interesn 1-year . The 52  = 0.08) . The 52  = 0.71) .Clearly the 52 mg-IUD/10–20 mcg-LNG-14t’s 1 in 1000 uterine perforation risk and invasive placement in the uterus are limitations that oral medications lack. Studies with IUDs cannot be completely blinded, and cannot have an innocuous a placebo control as other RCT can have. Normally an IUD string is left palpable in the vagina for IUD removal. The presence of the IUD string alerts the participant to the in utero IUD. Furthermore, any object placed in utero can exert a mass effect on the endometrium and uterus, therefore, a plastic IUD lacking hormonal or metallic active component is not inert. Incomplete participant and trial personnel blinding contributes to a performance bias . The 52 Clearly, the 52 mg-, 60 mg-, and frameless-IUD have long acting reservoir adherence advantages over daily oral progesterone for endometrial protection. While depo progestin precludes immediate treatment discontinuation, IUDs can be removed and progestin diffusion immediately halted. Oral progestin must be continuous to assure EH regression . NeverthLike progestins, metformin is anti-proliferative on the endometrium ,13,17. MPhosphoinositide 3-kinase (P13K) proto-oncogene serine/threonine protein kinase B (Akt) mammalian target of rapamycin  signaling pathway inhibition, mitogen-activated protein kinase (MAPK) inhibition, and glucose metabolism changes form part of metformin’s anti-proliferative effect ,11. MetfMetformin upregulates GLUT4 mRNA and protein which are normally decreased in PCOS in comparison to non-PCOS . Like prp = 0.002 [A case-controlled trial of 28 participants taking 850 mg twice daily for 7 to 30 days versus 12 participants not taking metformin found an average daily dose dependent 17.2% reduction in atypical EH and endometrioid EC Ki-67, 95% confidence interval (CI) −7.0%, −27.4%,  = 0.002 . This st = 0.002 . 2, p = 0.004 [p = 0.25 [A case series of five insulin resistant PCOS patients diagnosed by the Rotterdam criteria, who had Stage 1A Grade 1 EC treated with metformin 1000 mg daily and Cyproterone/Ethinyl Estradiol 2 mg/35 mcg 21 days per month for 6 months achieved 100% complete regression . This tr = 0.004 . HOMA-IRp = 0.25 . Cyprotep = 0.25 . Cyprotep = 0.25 . TherefoA pilot study compared megestrol acetate 160 mg daily to metformin 500 mg trice daily with megestrol acetate 160 mg daily for treatment of atypical EH . Only 16Metformin use is limited by its gastrointestinal adverse effect profile. Metformin is associated with abdominal pain, diarrhea, lactic acidosis, nausea and vomiting, and taste changes . MetformConsideration should be given to head-to-head trials of the 52 mg-IUD/10–20 mcg-LNG-14t and metformin combined with oral progesterone for EH and EC medical management. In vivo dose-escalation studies on metformin are needed to establish the optimal dose for EH treatment . MetformAromatase inhibitors that would reduce estrogen concentrations, gonadotrophin-releasing hormone agonists, and selective estrogen receptor modulators could be investigated for efficacy in preventing and/or treating EH and early stage EC . GLUT1 aProgestins have least three anti-proliferative mechanisms: anti-angiogenesis, estrogen receptor inhibition and IGF-1 inhibition . HoweverCyclic progestin has a 55.7 to 72% regression rate for EH, whereas continuous progestin has an 88 to 100% regression rate ,21. The p = 0.001 [p = 0.30 [Of note, with oral and intrauterine progestin therapy, premenopausal status and higher mean estrogen level are associated with EH recurrence,  = 0.001 . Conversp = 0.30 . While bHopefully, the feMMe trial, an international, phase II, RCT of the 52 mg-IUD/10–20 mcg-LNG-14t, metformin with the 52 mg-IUD/10–20 mcg-LNG-14t, and weight loss with metformin and the 52 mg-IUD/10–20 mcg-LNG-14t, for atypical EH and early stage EC patients will provide clarity on the effective of metformin concurrent with a progestin IUD . It is bIf the feMMe trial supports metformin use with progestin IUDs for medical management of EH and early stage EC, a combined metformin and LNG IUD may become a reasonable treatment modality. A combined metformin–LNG IUD may also have contraceptive and antiglycemic benefits for female, reproductive age diabetics. However, as the feMMe trial excludes oral progestins, unanswered questions will remain after the feMMe trial. The comparative effectiveness of oral progestins combined with metformin versus that of the IUDs combined with metformin for EH or early stage EC medical management remains to be answered.


Brucella species are the etiological agent of brucellosis in humans and animals. Here, we report the whole-genome sequence of Brucella melitensis strain CIIMS-BH-2, belonging to biovar 2. The draft assembly of CIIMS-BH-2 is 3.31 Mb in size, with 57.2% G+C content. Brucella species. Brucella spp. are Gram-negative, nonmotile, and facultative intracellular coccobacilli. Brucellosis in humans is characterized by undulant fever, arthritis, and gastrointestinal complications. It also causes a negative impact on livestock and public health , B. abortus (cattle), B. suis (swine), B. canis (dogs), B. ovis (sheep), B. neotomae (wood rats), B. ceti , B. pinnipedialis , B. microti (voles and foxes), B. inopinata (humans), B. papionism (baboons), and B. vulpis (foxes)  .Brucella spp. lack classical virulence factors, they have the ability to adapt to various environmental conditions  using version 2 chemistry. The sequencing of the isolate was performed on an Illumina HiSeq 2500 platform. The de novo assembly was carried out using SPAdes (version 3.10.1)  . The seqBrucella melitensis 16M. As predicted by RAST, the CIIMS-BH-2 genome consists of 3,216 open reading frames (ORFs), including 3,113 protein-coding sequences with 67 RNAs (The 16S rRNA gene sequence of CIIMS-BH-2 revealed 100% similarity to that of  67 RNAs . FurtherBrucella melitensis CIIMS-BH-2 genome sequence was deposited in the GenBank database under accession numbers CP025680 and CP025681.The


Volume 29, no. 3, Page 326–328, 2014Page 327, Legend for Fig. 1IncorrectFig. 1. Changes in viability (black line) and ATP levels (gray line) during carbon-starvation conditions.CorrectFig. 1. Changes in viability (gray line) and ATP levels (black line) during carbon-starvation conditions.


Bevacizumab addiction to triplet chemotherapy, according to FIr-B/FOx schedule, as first-line treatment in young-elderly metastatic colorectal CANCER (MCRC) patients may be more effective. Tailored treatments show worse clinical outcome in unfit patients.Elderly patients were clinically evaluated according to age and comorbidity  to select FIr-B/FOx regimen in fit or tailored treatments in unfit elderly. Limiting toxicity syndromes (LTS) were evaluated.KRAS genotype. G3-4 toxicities were diarrhea 21%, mucositis 11%, neutropenia 11%. LTS were 46%, significantly more multiple than single site. At 8 months follow-up, in 37 unfit patients: ORR 37%, PFS 7 months, OS 13 months. PFS was significantly different in KRAS wild-type compared to mutant patients, while not OS. PFS and OS were significantly worse in KRAS c.35 G > A compared to wild-type and/or other mutant.At 17 months follow-up, in 28 young-elderly patients treated with first line FIr-B/FOx: objective response rate (ORR) 79%, progression-free survival (PFS) 11 months, overall survival (OS) 21 months. Clinical outcome was not significantly different according to KRAS, and specifically c.35 G > A mutant genotype, may significantly affect clinical outcome in patients unfit for FIr-B/FOx.Careful decision-making process including evaluation of patient's fitness, and individual safety should be included to select FIr-B/FOx intensive first line regimen in young-elderly MCRC patients.  RAS genotype [KRAS wild-type patients [KRAS genotype) represents a major challenge in clinical management of MCRC patients.Different treatment options and lines of medical treatment in metastatic colorectal cancer (MCRC) patients are currently tailored according to fitness , comorbidities), metastatic extension (liver-limited (L-L) or other/multiple metastatic (O/MM)), and genotype –7. Firstgenotype , 7, 8, 1genotype , 10, 6. patients , 6. The MCRC patients are prevalently elderly but often under-treated in clinical practice, and usually underrepresented in clinical trials. They require a decision-making process combining the evaluation of fitness, according to co-morbidity, functional, and nutritional status , and selNo impact on PFS and OS was observed by age and/or comorbidities in patients treated with FOLFOX or FOLFIRI added or not to cetuximab . AdditioRecent retrospective analyses reported by our group, evaluating clinical outcome and safety of first-line FIr-B/FOx intensive regimen in fit young-elderly MCRC patients , 33, or et al. [2/w), OXP (32.5 mg/m2/w), associated to bolus plus continuous infusion 5-FU  880 mg/m2/w (88%)) failed to confirm significantly improved clinical efficacy compared to FOLFIRI in unresectable MCRC patients [HORG-FOLFOXIRI schedule proposed by Soug-lakos et al. , charact, OXP 32. mg/m2/w,patients , 5.In an unplanned subgroup evaluation of the phase III study, prognostic relevance of elderly status in both FOLFIRI and FOLFOXIRI treated patients was evaluated , and cliHORG/FOLFOXIRI intensive regimen showed a worse safety profile compared to FOLFIRI, with significantly more dose reductions and treatment delays; in elderly compared with younger patients, significantly more dose reductions , treatment delays , particularly due to toxicity were reported . In the FOLFIRI arm, dose reductions  and treatment delays were not different. However, there was no significantly different rDI of drugs in elderly compared with younger patients, in both FOLFOXIRI and FOLFIRI arms.Grade 3-4 diarrhea was significantly more prevalent in elderly, compared to younger patients, in both chemotherapy regimens  . Moreove2, 12h-timed-flat-infusion, 2 days weekly; CPT-11 160 mg/m2/BEV 5 mg/kg or OXP 80 mg/m2, weekly alternating. Cumulative Index Rating Scale (CIRS) was used to evaluate the comorbidity status, and only patients with primary and intermediate CIRS stage were enrolled [We retrospectively evaluated consecutive young-elderly patients 65 to 75 years enrolled in the previously reported phase II trial  and in tenrolled . Primaryenrolled . LTS werKRAS wild-type patients, ORR was 92%, liver metastasectomies 23% (50% in L-L patients), median PFS 14 months (4-78+ months), median OS 38 months (8-78+ months). Among 13 KRAS mutant patients, ORR was 77%, liver metastasectomies 15%, (20% in L-L patients), median PFS 7 months (3-69+ months), median OS 19 months (6-69+ months). Neither PFS nor OS were significantly different in KRAS wild-type compared with mutant patients, according to log-rank test. No BRAF mutations were detected.Young-elderly patients were 28 (42%) among overall MCRC patients enrolled fitting for FIr-B/FOx intensive treatment, according to inclusion criteria, WHO PS 0 89%, CIRS primary/intermediate. At a median follow-up of 17 months, ORR was 79%, liver metastasectomies 18% (37.5% in L-L patients), median PFS 11 months (3-78+), median OS 21 months (6-78+) . Patients’ distribution according to age and comorbidities was: young-elderly 22%, old-elderly 54%; CIRS stage primary 11%, intermediate 40%, secondary 42%. Medical treatment regimens were tailored according to age and comorbidity status in the individual patients. Eighteen patients (49%) were treated with triplet regimens; 15 patients (40%) with doublet regimens; 4 patients (11%) with mono regimens; 3 underwent up-front surgery.At a median follow-up of 8 months, ORR was 37%, median PFS was 7 months (1-13+), median OS 13 months (1+-23+). Among patients treated with triplet regimens, ORR was 37.5%, median PFS 8 months (3-12), median OS 12 months (3-23+ months) , median OS 13 months (1+-23+ months). Among 12 KRAS mutant patients evaluable for activity, ORR was 25%, median PFS 6 months (1-11 months), median OS 8 months (3-18 months). A significantly different PFS (p = 0.043), but not OS, was reported in KRAS wild-type compared with mutant patients. Significantly worse PFS and OS were reported in c.35 G > A KRAS mutant compared to wild-type , and to other mutant patients .Among 14 KRAS wild-type patients, demonstrated clinical outcome and safety profile equivalent to younger patients [Fit young elderly patients, PS<2, treated with triplet regimens consisting of chemotherapeutic drugs, or BEV addiction to doublets, or doublets plus EGFR-inhibitors in patients , 5, 8, 1patients , 34 obtaRetrospective analysis of randomized clinical trials showed that doublets CPT-11, or OXP, added to fluoropyrimidin in older patients eligible for clinical study reported ORR 18-59.4%, PFS 4.9-10.0 months and OS 8.5-20.7 months , 32, 30.Published studies showed that limiting toxicities were not significantly different in elderly patients treated with 5-FU or CPT-11 –16, sligKRAS wild-type compared to mutant patients. More, significantly worse clinical outcome (PFS and OS) may be influenced by KRAS c.35 G > A mutant genotype, compared to wild-type and/or other mutant, confirming that KRAS genotype, and specifically c.35 G > A mutant, confers different biological aggressiveness [KRAS genotype may significantly explain different PFS, and c.35 G > A KRAS mutant a significantly worse PFS and OS, compared to wild-type and other mutant.In clinical practice, selection of patients eligible for intensive medical treatment and to achieve optimal activity and clinical outcome could be performed by the evaluation of age, PS, CIRS and careful monitoring of individual safety using LTS Table . Patientsiveness –41, lessKRAS genotype, and specifically the prevalent KRAS c.35 G > A mutant status, that can discriminate significantly different clinical outcome particularly in unfit MCRC patients.We revised intensive medical treatment consisting of triplet chemotherapy regimens or more intensive triplet chemotherapy plus anti-angiogenic drug, bevacizumab, in elderly MCRC patients, that we previously developed, discussed compared with tailored medical regimens selected for unfit MCRC in clinical practice , 7, 32,


The correct name is: Victor W. M. van Hinsbergh. The correct citation is: Nauta TD, Duyndam MCA, Weijers EM, van Hinsbergh VWM, Koolwijk P (2016) HIF-2α Expression Regulates Sprout Formation into 3D Fibrin Matrices in Prolonged Hypoxia in Human Microvascular Endothelial Cells. PLoS ONE 11(8): e0160700. doi:


Primulina cardaminifolia Yan Liu & W.B. Xu (Gesneriaceae), a distinct new species with imparipinnate leaves, is described and illustrated from a limestone valley in Guangxi Zhuangzu Autonomous Region, China. To assure its generic placement and phylogenetic affinity, phylogenetic analyses were performed using DNA sequences of nuclear ITS and chloroplast trnL-F intron spacer region. Additionally, somatic chromosome number was counted and pollen stainability was tested.Primulina; however, two phylogenetically distinct ITS sequence types were detected, suggesting a probable hybrid origin. Its pollen stainability is 100% and its chromosome number, 2n = 36, is congruent with all known counts of diploid species of the genus.Phylogenetic analyses support its placement in Primulina cardaminifolia and suggest that it could have derived from homoploid hybrid speciation. Color plates, line drawings and a distribution map are provided to aid in identification.All available data support the recognition of the new species The online version of this article (doi:10.1186/1999-3110-54-19) contains supplementary material, which is available to authorized users. Chirita D. Don  Mich. Möller & A. Weber] made without support from molecular data by Weber et al. .Although the new generic circumscription reflects better the evolutionary relationships and ecological preferences of the genus, diagnostic characters of  rosette  and the r et al.  was recePrimulina, which is described and illustrated here. Chromosome count of the new species is in agreement with those reported for Primulina in Christie et al.  has been deposited in HAST. Root tips were gathered and pretreated in 2 mM 8-hydroxyquinoline at 15–18°C for about 6 h and fixed overnight in an ethanol-acetic acid solution (3:1) below 4°C. The chromosomes were stained and macerated in 2% acetic orcein with 1 N hydrochloric acid (10:1). Classification of chromosome complement based on centromere position at mitotic metaphase followed Levan et al.  and cycle-sequenced using the T7 and SP6 promoter primer pairs.DNA sequences of the nuclear ribosomal internal transcribed spacers (ITS) and the chloroplast u et al. . BecausetrnL-F regions as separated matrices. Primulina pinnata (W.T. Wang) YinZ. Wang, which was not included in Xu et al.  YinZ. Wang, and Didymocarpus podocarpus C.B. Clarke chosen as outgroups based on recent phylogenetic analyses  and maximum likelihood (ML) criteria implemented in MEGA5. MP trees were searched using the Tree-Bisection-Reconnection (TBR) search option with the initial trees setting at 50, MP search level setting at 5, and maximum number of trees setting at 2000. Clade supports were calculated based on 100 bootstrap resamplings . ML trees were reconstructed using the nearest-neighbor-interchange (NNI) method with all site used and the initial tree automatically selected under the model(s) selected by MEGA5. Clade supports of ML analysis were evaluated based on 100 bootstrap resamplings .For phylogenetic analyses, matrices of Xu et al.  were adou et al.  because Wei-Bin Xu & Yan Liu 08050 . 碎米薺葉報春苣苔 Figures Yan Liu & W.B. Xu, sp. nov.—TYPE: CHINA. Guangxi Zhuangzu Autonomous Region, Laibin Shi (City), Fenghuang Zhen (Township), alt. 280 m, on moist limestone rock face in a valley, 14 July 2008, Primulina cardaminifolia Yan Liu & W.B. Xu resembles Primulina pinnata (W.T. Wang) YinZ. Wang in having imparipinnate leaves, but is clearly distinct from this species by the ovate-cordate terminal leaflet of 3–7 × 3–6.5 cm, 1 or 2 pairs of broadly ovate to sub-ovate lateral leaflets, 1–3-branched cymes with 3 to 10-flowers, and the entire-margined calyx lobes with acuminate apex.Herbs perennial. Rhizome subterete, 3–5 mm across. Leaves 5–7, in basal rosette, imparipinnate, 10–20 cm long, papery when dry; petiole subterete, 6–12.5 cm long, densely pubescent; terminal leaflet ovate-cordate, 3–7 × 3–6.5 cm, apex obtuse, base cordate, margin repand to irregularly pinnately lobed, densely pubescent on both surfaces; lateral leaflets 1 or 2 pairs, opposite or alternate, broadly ovate to rotund, 1–3 × 1–3 cm, margin repand to irregularly pinnately lobed, densely pubescent on both surfaces, petiolules short, 2–10 mm long. Cymes 2–4, axillary, 1–3-branched, 3–10-flowered; peduncle 4–8 cm long, 1–2 mm in diam., densely pubescent; bracts 2, opposite, linear-lanceolate, 7–8 × 2–3 mm, margin entire, pubescent; pedicel 4–7 mm long, densely pubescent. Calyx 5-parted to base, lobes lanceolate-linear, 8–12 × 2–3 mm, apex acuminate, outside pubescent, inside glabrous, margins entire. Corolla white to pale purple, 3.2–3.5 cm long, outside glandular-pubescent, inside sparsely puberulent, with 2 pale-yellow stripes; corolla tube 2.1–2.3 cm long, 8–12 mm in diam. at the mouth, 2.5–3 mm in diam. at the base; limb pale purple, distinctly 2-lipped; adaxial lip 2-parted to over the middle, lobes oblong, 8–10 × 6–7 mm; abaxial lip 3-lobed to over the middle, lobes oblong, 12–13 × 4–5 mm; stamens 2, adnate to 1.5 cm above the corolla tube base; filaments linear, ca. 1.2 cm long, geniculate above the base, sparsely glandular-puberulent; anthers ca. 3 mm long, ca. 1.5 mm wide, dorsifixed, glabrous; staminodes 2, ca. 5 mm long, apex capitate, glabrous, adnate to ca. 6 mm above the base of corolla tube. Disc ring-like, ca. 1 mm in height, margin repand, glabrous. Pistil 2.5–2.8 cm long, ovary 7–8 mm long, ca. 1.5 mm across, puberulent; style 1.5–1.8 cm long, ca. 0.6 mm across, puberulent; stigma obtrapeziform, ca. 2 mm long, apex 2-lobed. Capsules not seen.CHINA. Guangxi Zhuangzu Autonomous Region, Laibin Shi, Fenghuang Zhen, 3 July 2008, Wei-Bin Xu & Yan Liu 08040 (IBK); same locality, 8 Sep 2008, Wei-Bin Xu & Kuo-Fang Chung 08472 (IBK), Shin-Ming Ku et al. 2035 (HAST); same locality, 28 June 2007, Hong-Jin Wei & Wei-Bin Xu 07244 (IBK).Primulina cardaminifolia is extremely rare, currently known only from the type locality in Laibin Shi, Guangxi Zhuangzu Autonomous Region, China .The specific epithet is derived from the leaves resembling those of the genus Primulina cardaminifolia resembles Primulina pinnata (W.T. Wang) YinZ. Wang  YinZ. Wang  and 634 (P. cardaminifolia-B) bp, respectively, while only one cpDNA sequence type was detected in the species. With the addition of these two ITS sequences, the ITS matrix contained 28 accessions of 675 aligned positions, of which 182 (26.96%) were parsimoniously informative. Based on the Kimura 2-parameter model using a discrete Gamma distribution (K2 + G) with 5 rate categories selected by the corrected Akaike Information Criterion (AICc) implemented in MEGA5, a single ML tree (log likelihood = −4114.9869) was recovered  and low  supports in ITS and cpDNA datasets, respectively . Primulina cardaminifolia was collected from the type locality [S.M. Ku 2035(HAST)].GenBank accession numbers: Species: (ITS/Didymocarpus podocarpus C.B. Clarke: (DQ912688/FJ501514); Petrocodon dealbatus Hance: (FJ501358/FJ501537); Petrocodon scopulorum (Chun) YinZ. Wang [=Tengia scopulorum Chun]: (GU350637/GU350669); Primulina bipinnatifida (W.T. Wang) Yin Z. Wang [=Chiritopsis bipinnatifida W.T. Wang]: (DQ872842/DQ872806); Primulina cardaminifolia Yan Liu & W.B. Xu: ; Primulina cordifolia (D. Fang & W.T. Wang) YinZ. Wang [=Chiritopsis cordifolia D. Fang & W.T. Wang]: (DQ872845/DQ872803); Primulina dryas (Dunn) Mich. Möller & A. Weber [=Chirita sinensis Lindl.]: (FJ501348/FJ501524); Primulina gemella (D. Wood) YinZ. Wang [=Chirita gemella D. Wood]: (FJ501345/FJ501523); Primulina glandulosa  Yin Z. Wang : (DQ872841/DQ872804); Primulina heterotricha (Merr.) YinZ. Wang [=Chirita heterotricha Merr.]: (DQ872826/DQ872816); Primulina linearifolia (W.T. Wang) YinZ. Wang [=Chirita linearifolia W.T. Wang]: (DQ872834/DQ872810); Primulina longgangensis (W.T. Wang) YinZ. Wang [=Chirita longgangensis W.T. Wang]: (FJ501347/AJ492290); Primulina luochengensis (Yan Liu & W.B. Xu) Mich. Möller & A. Weber [=Wentsaiboea luochengensis Yan Liu & W.B. Xu]: (HQ633046/HQ632949); Primulina minutimaculata (D. Fang & W.T. Wang) YinZ. Wang [=Chirita minutimaculata D. Fang & W.T. Wang]: (DQ872828/DQ872815); Primulina multifida B. Pan & K.F. Chung: (JX507031/JX506756); Primulina ophiopogoides (D. Fang & W.T. Wang) YinZ. Wang [=Chirita ophiopogoides D. Fang & W.T. Wang]: (DQ872829/DQ872814); Primulina pinnata (W.T. Wang) YinZ. Wang [Chirita pinnata W.T. Wang]: (FJ501349/FJ501526); Primulina pinnatifida (Hand.-Mazz.) Yin Z. Wang [=Chirita pinnatifida (Hand.-Mazz.) B.L. Burtt]: (FJ501350/FJ501527); Primulina pseudomollifolia W.B. Xu & Yan Liu: (JX506869/JX506759); Primulina pteropoda (W. T. Wang) Yan Liu [=Chirita pteropoda W.T. Wang]: (DQ872827/DQ872817); Primulina repanda var. guilinensis (W.T. Wang) Mich. Möller & A. Weber [=Chiritopsis repanda var. guilinensis W.T. Wang]: (DQ872846/DQ872808); Primulina spadiciformis (W.T. Wang) Mich. Möller & A. Weber [=Chirita spadiciformis W.T. Wang]: (FJ501346/AJ492291); Primulina spinulosa (D. Fang & W.T. Wang) YinZ. Wang [=Chirita spinulosa D. Fang & W.T. Wang]: (DQ872830/DQ872813); Primulina tabacum Hance: (FJ501352/AJ492300); Primulina weii Mich. Möller & A. Weber : (DQ872832/DQ872811); Primulina wentsaii (D. Fang & L. Zeng) YinZ. Wang [=Chirita wentsaii D. Fang & L. Zeng]: (DQ872831/DQ872812).


Listeria monocytogenes is a Gram-positive, rod-shaped, non-spore-forming bacterium which is an important foodborne bacterial pathogen for humans worldwide with high mortality rates. Here, we report a 2,964,284-bp draft genome sequence of Listeria monocytogenes strain ATCC 7644 (American Type Culture Collection). Listeria monocytogenes is a Gram-positive, rod-shaped, non-spore-forming bacterium and an important foodborne bacterial pathogen for humans worldwide with high mortality rates  isolated from a human was obtained from ATCC  and stored at −80°C. Genomic DNA was isolated from overnight cultures grown at 37°C on brain and heart infusion agar using the Qiagen DNeasy blood and tissue DNA purification kit . Three sequencing libraries were constructed from three independent cultures/DNA extractions using the Nextera XT DNA sample preparation kit , and library quality was analyzed using a BioAnalyzer . Libraries were sequenced with the Illumina MiSeq platform  using v3 reagent kits. A total of 2,889,385 300-bp paired-end reads were generated.http://jgi.doe.gov/data-and-tools/bbtools/). Preprocessed reads were assembled using SPAdes v3.10.1 , and overlapping pairs were merged (bbmerge) using the bbtools software suite ( v3.10.1  and poli v3.10.1 . Assemblh v1.1.1 , which ph v1.1.1 . Two of NGZM00000000. The version described in this paper is the first version, NGZM01000000.This whole-genome shotgun project has been deposited in DDBJ/ENA/GenBank under the accession no.


P = 0.03, I2 = 0%; CC vs. GG: OR = 0.59, 95% CI = 0.36–0.97, P = 0.04, I2 = 0%; GC+CC vs. GG: OR = 0.61, 95% CI = 0.37–0.99, P = 0.05, I2 = 0%). In summary, current evidence indicates that the microRNA-608 rs4919510 G>C polymorphism maybe an important factor of DSCs susceptibility, especially in Caucasian population.Previous epidemiologic studies have revealed a possible association between microRNA-608 rs4919510 G>C polymorphism and digestive system cancers (DSCs) risk, but the results were not consistent. We therefore performed an updated meta-analysis to explore the association between microRNA-608 rs4919510 G>C polymorphism and DSCs risk. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to assess the relationship between the microRNA-608 rs4919510 G>C polymorphism and DSCs risk. Heterogeneity, cumulative analyses, sensitivity analyses, and publication bias were also conducted to examine the statistical power. Eight published articles with nine independent case-control studies involving 10,836 individuals were included in this meta-analysis. Overall, no significant association was found between microRNA-608 rs4919510 G>C polymorphism and DSCs risk in general populations. But some significant protective effects were observed in the subgroup of Caucasian population group in three genetic models (C vs. G: OR = 0.82, 95% CI, 0.68–0.99,  Digestive system cancers (DSCs), which comprise esophageal cancer (EC), gastric cancer (GC), hepatocellular cancer (HC), pancreatic cancer (PC) colorectal cancer (CRC) and other solid carcinoma, is one of the most common malignancies with increasing incidence and mortality worldwide  are the most common type of genetic variation in people. SNPs alter gene function and/or expression, consequently affecting downstream biologic pathways and increasing cancer risk  statement  were searched for relevant studies that focused on the association between microRNA-608 rs4919510 G>C polymorphism and DSCs risk from inception up to January 1, 2018. Only studies that were written in English and Chinese were included. Moreover, the bibliographies of the collected studies and relevant reviews were retrospected to identify additional articles. The following search terms and strategy was used :#1 microRNA 608#2 microRNA-608#3 mir 608#4 mir-608#5 rs4919510#6 #1 OR #2 OR #3 OR #4 OR #5#7 polymorphism#8 variant#9 mutation#10 #7 OR #8 OR #9#11 cancer#12 tumor#13 neoplasm#14 #11 OR #12 OR #13#15 #6 AND #10 AND #14Studies were included based on the following criteria: (1) only case-control studies that focusing on the association between the microRNA-608 rs4919510 G>C polymorphism and DSCs risk; and (2) studies had to provided sufficient frequency data on the genotype distribution to evaluate the crude odds ratios (ORs) and 95% confidence intervals (CIs); and; (3) studies had to be published only in English and Chinese; and (4) only publications with the largest or most recently updated sample data were included when there were some overlapping or duplicate publications on the same theme.Two researchers (Li and Song) independently reviewed and extracted relevant information from all included studies, including name of the first author, publishing date, country or region where the study was conducted, race, control design, sample sizes of the cases and controls, frequency data of genotype distribution, genotyping method, Hardy-Weinberg equilibrium (HWE) assessment in controls, and cancer type. Quality evaluation of the included studies was performed by the two researchers using the modified Newcastle-Ottawa scale (NOS). The scores ranged from 0 points (worst) to 11 points (best) Table ; studiesQ-test and I2 statistical method , co-dominant models (GC vs. GG and CC vs. GG), dominant model (GC+CC vs. GG), and recessive model (CC vs. GG+GC). Heterogeneity between the included studies was calculated using Cochran's A total of 164 articles were collected. The study selection process was presented in Figure P = 0.98, I2 = 53.4%; GC vs. GG: OR = 1.07, 95% CI = 0.93–1.24, P = 0.35, I2 = 43.0%; CC vs. GG: OR = 1.01, 95% CI = 0.83–1.24, P = 0.90, I2 = 56.4%; GC+CC vs. GG: OR = 1.05, 95% CI = 0.89–1.23, P = 0.58, I2 = 56.4%,  .Publication bias was examined with Begg's test and no apparent asymmetry of the funnel plot was found Figure . These rMicroRNAs is a family of small, noncoding, evolutionarily conserved endogenous RNAs consisting of 22 nucleotides that can regulate gene expression through a post-transcriptional pathway binding to the 3′-UTR of target mRNAs , and the results were calculated without gene-gene and gene-environment risk factors, leading to a failure to interpret the potential interaction mechanisms. Third, almost all of the included studies focused on Asian and Caucasian populations, which would restrict the application of our results to other populations.In summary, our meta-analysis demonstrated that the microRNA-608 rs4919510 G>C polymorphism might play an important role in DSCs susceptibility. Further studies in different races and regions with larger population sizes are needed to confirm our findings.X-FL, J-KS, and Y-MN conceived the study. X-FL, J-KS, and J-WC searched the databases and extracted the data. J-KS, Y-QZ, and ML analyzed the data. X-FL, J-KS, and J-WC wrote the draft of the paper. JZ and Y-MN reviewed the manuscript. All the authors approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Molecular classifications of breast cancer have resulted in improved treatments. However, treatments for triple negative breast cancer (TNBC) are lacking. Analysis of molecular targets for TNBC is a priority. One potential candidate is androgen receptor (AR) phosphorylation. This study assessed the role of AR phosphorylation at ser81/ser515 and their two upstream effectors, cyclin-dependent kinase 1 (pCDK1) and extracellular-regulated kinase 1/2 (pERK1/2) in 332 ductal breast cancer patients by immunohistochemistry.p = 0.038), decreased size (p = 0.001), invasive grade (p < 0.001), necrosis (p = 0.003), b-lymphocytes (p = 0.020), molecular subtype (p < 0.001) and estrogen receptor (ER)/progesterone receptor (PR)-status (p < 0.001). The cohort was therefore stratified into ER+ve and ER-ve patients. In ER+ve tumours, pERK1/2 combined with AR-515 associated with improved CSS (p = 0.038), smaller size (p = 0.004), invasive grade (p = 0.001), decreased b-lymphocytes (p = 0.013) and increased plasma cells (p = 0.048). In contrast, in TNBC patients, phosphorylation of AR-515 associated with poorer CSS (p = 0.007). pERK1/2 combined with AR-515 associated with decreased inflammation (p = 0.003), increased tumour stroma (p = 0.003) and tumour budding (p = 0.011), with trends towards decrease CSS (p = 0.065) and macrophage levels (p = 0.093).pERK1/2 combined with AR-515 associated with improved cancer-specific survival (CSS, In Conclusions, AR-515 may be an important regulator of inflammation in breast cancer potential via ERK1/2 phosphorylation. AR-515 is a potential prognostic marker and therapeutic target for TNBC. Approximately 1.7 million cases of breast cancer are diagnosed every year. About 522,000 die of breast cancer yearly making it the 5th most common cause of cancer death globally . In the The majority of breast cancers comprise of luminal A and luminal B subtypes. These subtypes display oestrogen (ER) and/or progesterone receptors (PR) rendering these cancer cells susceptible to endocrine therapies e.g. anti-oestrogen therapies (Tamoxifen) , 8. In HIt has been shown that TNBC patients displaying pathological complete response (pCR) had better outcomes in terms of progression-free survival than patients who had not achieved pCR . pCR is Although numerous studies on AR have been published, the exact role of AR as tumour-promoting or anti-tumorigenic remains undecided. It is known that AR signalling plays a role in regulating cancer cell proliferation and apoptosis –19. AR aIt was also reported that when Tamoxifen-resistant breast cancer cells that expressed high levels of AR were exposed to bicalutamide (anti-androgen), cell growth was shown to decrease as well as reversing Tamoxifen resistance , indicatThe MAPK/ERK signalling cascade is heavily involved in the regulation of cancer cell proliferation, growth and differentiation. The stimulation of this pathway leads to the activation of a cascade of kinases , which ultimately leads to the activation of ERK1/2, which is associated with poor prognosis in breast cancer . One funin vitro kinase assays demonstrated an increase in ser-308 phosphorylation by CDK1 is responsible for Serine–81 phosphorylation [In contrast, AR phosphorylation at ser-81 is the prototypical site for dihydrotestosterone (DHT) activation, which is thought to be facilitated by activated Cdk1, as observed by Chen et al where Cdk1 inhibitors decreased ser-81 phosphorylation of AR in LNCaP cells and similarly decreased AR protein expression and transcriptional activity. Whereas transfected Cdk1 stimulated AR phosphorylation at ser-81 and increased AR protein expression . This surylation . Cdk1 phrylation .This study aims to further understand the role of androgens and AR activation in breast cancer by determining its effect on patient survival. This is followed by the determination of the role of upstream pathways (i.e. Cdk1 and ERK1/2 pathways) on AR activation and their effects on cancer specific survival (CSS). We aim to confirm that the phosphorylation of AR at either ser-515 or ser-81 is a surrogate for AR activation and assess if the pathways responsible for this phosphorylation are potential targets for therapeutic intervention.n = 332, Figure Only patients with staining for AR, AR-81 and AR-515 were included in the analysis (p = 0.842) nor AR-515 (p = 0.563) were associated with CSS alone. Therefore, upstream effectors of AR phosphorylation were investigated. AR-81 significantly correlated with nuclear CDK1 (p < 0.001) and AR-515 correlated with nuclear pERK1/2 (p < 0.001) as expected. However, AR-81 still showed no associations even when combined with CDK1 in the nucleus (p = 0.344). In contrast, AR-515 was significantly associated with improved CSS when combined with ERK1/2 phosphorylation in the nucleus . When nuclear pERK1/2 was assessed alone it was also significantly associated with improved CSS (p = 0.004). Therefore, further analysis was confined to nuclear pERK1/2 and AR-515.The relationship between nuclear AR activation and CSS is shown in Table p = 0.001), lower grade (p < 0.001), ER/PR+ve tumours (p < 0.001), molecular subtype (p < 0.001), lower necrosis (p = 0.003), weak Klintrup-Makinen (KM) grade (p = 0.004) and lower b-lymphocyte levels (p = 0.020). There was also a trend towards decreased HER2 (p = 0.096) and lower lymphovascular invasion (p = 0.099). When pERK1/2 was combined with AR-515 significant associations were still seen with smaller size (p = 0.017), lower grade (p < 0.001), ER+ve tumours (p = 0.013), PR+ve tumours (p = 0.008), decreased HER2 (p = 0.041), molecular subtype (p < 0.001), lower necrosis (p = 0.006), weak KM grade (p = 0.005) and lower b-lymphocytes (p = 0.023). Significant associations were now also seen with decreased proliferation (p = 0.009), lymphovascular invasion (p = 0.017) and increased tumour budding (p = 0.047).The relationship between pERK1/2, AR-515, clinicopathological characteristics, and local inflammatory response is presented in Table p = 0.008). As pERK1/2 and AR-515 strongly correlated in ER+ve patients (p < 0.001) they were combined. However, when combined the survival effect was weakened (p = 0.038), suggesting that pERK can also act independently of AR-515 in ER+ve breast cancers. When ER+ve patients where stratified further, there was no difference in CSS between luminal A and luminal B subtypes for pERK1/2 alone or combined with AR-515 therefore further analysis was carried out in all ER+ve tumours (n = 228).As pERK1/2 plus AR-515 associated with molecular subtype, we firstly stratified patients into ER+ve and ER-ve tumours . pERK1/2p = 0.004), lower grade (p = 0.001), molecular subtype (p = 0.024), decreased proliferation (p = 0.006), lower b-lymphocytes (p = 0.013) and increased plasma cells (p = 0.048). There was also a trend towards decreased necrosis (p = 0.096). When pERK1/2 was combined with AR-515 significant associations where still seen with lower grade (p = 0.002), molecular subtype (p = 0.001), decreased proliferation (p = 0.001), lower b-lymphocytes (p = 0.006) and increased plasma cells (p = 0.023). Significant associations were now also seen with decreased necrosis (p = 0.009) and lymphovascular invasion (p = 0.029).The relationship between pERK1/2, AR-515, clinicopathological characteristics, and local inflammatory response in ER+ve tumours is presented in Table p = 0.023) may be due to differences between the two ER-ve subtypes. Therefore, ER-ve patients were stratified into HER2-type and TNBC to further investigate these effects (p = 0.010), however no significant effect on CSS was observed for pERK1/2 alone (p = 0.534) and only a trend towards poorer outcome was seen when combined with AR-515 (p = 0.065). However, high levels of AR-515 combined with total AR, representing full AR activation, associated with a significantly poorer outcome  and no significant associations were seen in these patients. However, there was a trend towards improved patient survival with AR-515 phosphorylation (p = 0.056) alone and when combined with pERK1/2 (p = 0.076), confirming our hypothesis that these two ER-ve subtypes respond differently to AR-515 phosphorylation.It was observed in patients with ER-ve breast cancer that high pERK1/2 plus AR-515 had a trend towards an effect on survival rate. It was hypothesised that the loss of significance and weaker correlation between pERK1/2 and AR-515 in ER-ve patients ( effects . In patip = 0.033), lower KM grade (p = 0.004), increased tumour-stroma percentage (TSP) (p = 0.033) and increased tumour budding (p = 0.007). When combined with pERK1/2 associations were still seen with lower KM grade (p = 0.003), increased TSP (p = 0.003) and increased tumour budding (p = 0.011). Trends towards association were also seen for increased age (p = 0.075) and decreased macrophages (p = 0.093).Due to the detrimental effect of AR-515 phosphorylation in TNBC, associations with clinicopathological characteristics were assessed Table . Full acp = 0.002), invasive grade (p = 0.013), lymph node involvement (p = 0.009), necrosis (p < 0.001), proliferation index (p < 0.001), lymphatic invasion (p < 0.001), cytotoxic t-lymphocytes (p = 0.005), TSP (p = 0.009), tumour budding (p = 0.001) and angiogenesis (p = 0.043) were significantly associated with poorer CSS. Lymphovascular invasion (p = 0.054) and b-lymphocytes (p = 0.064) trended towards significance.For ER+ve tumours, since pERK1/2 plus AR-515 had significant associations with improved patient survival, these factors were taken into univariate and multivariate analysis (Table p = 0.036), proliferation index (p = 0.014), cytotoxic t-lymphocytes (p = 0.001), and TSP (p = 0.033) were independent prognostic factors, with lymphatic invasion (p = 0.060), tumour budding (p = 0.084), angiogenesis (p = 0.051) and pERK1/2 (p = 0.073) showing trends towards independence. However, pERK1/2 plus AR-515 does not appear to be independently prognostic in these patients (p = 0.682).On multivariate survival analysis including pERK1/2 plus AR-515; necrosis (p = 0.022), lymph node involvement (p < 0.001), necrosis (p = 0.030), lymphatic invasion (p = 0.001), lymphvascular invasion (p < 0.001), macrophages (p = 0.011) and cytotoxic t-lymphocytes (p = 0.047) were significantly associated with poorer CSS. Age (p = 0.055) and plasma cells (p = 0.055) trended towards significance.For TNBC, since AR-51 plus total AR had significant associations with poorer patient survival, these factors were taken into univariate and multivariate analysis (Table 7). On univariate analysis, size (p = 0.006), lymphovascular invasion (p = 0.002), and macrophages (p = 0.002) were independent prognostic factors. However, AR-515 plus total AR (p = 0.957) nor AR alone (p = 0.801) appear to be independently prognostic in these patients.On multivariate survival analysis including AR-515 plus total AR; lymph node involvement . The study was approved by the Research Ethics Committee of the West Glasgow University Hospitals NHS Trust.Patients presenting with invasive ductal breast cancer at Glasgow Royal Infirmary, Western Infirmary and Stobhill Hospital between 1995 and 1998 with formalin-fixed paraffin embedded blocks of the primary tumour available for evaluation were studied  were retrieved from the routine reports. Tumour grade was assigned according to the Nottingham Grading System. ER and PR status were assessed on tissue microarrays (TMAs) using immunohistochemistry (IHC) with Dako  ER antibody and Leica  PR antibody and scored according to the American Society of Clinical Oncology and College of American Pathologists guidelines with a cut-off value of 1% positive tumour nuclei . Her2 stFull-section haematoxylin and eosin (H&E) slides for the 474 patients were used to score local inflammatory infiltrate according to Klintrup-Makinen (KM) criteria. KM scoring of slides was carried out as previously described. Briefly, tumours were scored on four-point scores based on appearances at the tumour invasive margin. A score of 0 signified that there were no inflammatory cells at tumour's invasive margin; score 1 indicated a mild and patchy inflammatory cells; score 2 denoted a prominent band-like inflammatory reaction at the invasive margin; and score 3 revealed a florid cup-like inflammatory infiltrate at the invasive edge , 44. IndFull-section H&E slides were also used to score the tumour stroma percentage (TSP) as previously reported . BrieflyLymphatic and blood vessel (lymphovascular) invasion were assessed, on 2.5-μm thick sections, using IHC staining with the lymphatic endothelial marker D2-40  diluted 1:100 and vascular endothelial marker Factor VIII  diluted 1:100 as previously described . Ki67 prThe molecular subtypes were defined as follows: Luminal A: oestrogen (ER) and/or progesterone receptor (PR) positive, Her-2 negative or low proliferative index (< 15%); Luminal B: hormone receptor positive, Her-2 positive or high proliferative index (> 15%); Her-2 subtype: Her-2 positive and hormone receptor negative, any proliferative index; and triple negative: Her-2 negative, hormone receptor negative, any proliferative index.2O2. Sections were further blocked using 5% horse serum (10% casein for pERK1/2) in Tris-buffered saline (TBS). Antibodies for pCdk1 (Abcam), pERK1/2 , AR-81 , AR-515  were incubated overnight at 4°C diluted in Dako antibody diluent at 1:150 (Dako UK Ltd.) Total AR antibody (Dako UK Ltd.) was incubated overnight at 4C at 1:4000. Bound antibody complex was visualised using EnVision plus kit (Dako UK Ltd.) followed by 3,3-diaminobenzidine tetrahydrochloride . Nuclei were counterstained with haematoxylin and blued with Scots Tap Water Substitute, dehydrated through graded alcohol and xylene and mounted with Di-N-Butyl Phthalate in xylene (DPX).Antibody validation for AR-81, AR-515, pERK1/2 and pCDK1 was carried out as previously described , 26. ImmTMAs were scanned using the Hamamatsu NanoZoomer  at × 20 magnification, and visualization was carried out using Slidepath Digital Image Hub, version 4.0.1 .Only nuclear expression was scored as this represents the active form of each protein. Tissue staining intensity was scored by two blinded independent observers using a weighted histo-score (H-score) method , 49. Then = 332). Cut-off values for high or low protein expression were determined by the median of the average values of the intensity of staining. The relationships between variables were assessed using contingency table analysis with the X2 test for linear trend. Correlations coefficients were analysed using a Spearman's rho. Kaplan– Meier curves with log rank analysis was used to examine the effect of protein expression on CSS. Univariate survival analysis was performed using Cox proportional hazards regression. Variables with P-value of < 0.05 were entered into a multivariable model using a backwards conditional method for all patients and triple negative patients. All statistical analyses were two-sided and significance defined as P-value < 0.05. All statistical analysis was performed using the SPSS software version 22 .Only patients with a score for AR, AR-81 and AR-515 were included in the analysis (


The absolute configurations of four terpenes were determined based on their optical rotary powers. Incubation experiments with  R)-(+)-limonene is found in citrus fruits. Cineol is one of the main constitutents of eucalyptus oil and (+)-carvone is present in caraway. Some famous sesquiterpenes are α-humulene from hops, α-patchoulene from patchouli oil, and β-cedrene from juniper D = +120) , wh, whStrepades ago , the fultructure . The sig carbons . The HMB = +120) , while i2+ cofactor to which in turn the substrate’s diphosphate portion is bound D24 = +7.1 . This points to the same enantiomer as reported from the heartwood of Cryptomeria japonica D22 = −51.3 . The opposite enantiomer was reported from Eucalyptus macarthuri ([α]D = +30.5) + calcd for C15H24+, 204.1873; found, 204.1872; IR (diamond ATR) : 2964 (m), 2925 (m), 2873 (m), 1670 (w), 1447 (w), 1412 (w), 1377 (w), 1259 (s), 1090 (s), 1016 (s), 866 (w), 797 (s), 700 (w), 662 (w) cm−1.I = 1483 ); MS (EI(+)-T-Muurolol (2): GC (HP 5): I = 1640 (literature (HP 5): I = 1640 + calcd for C15H26O+, 222.1978; found, 222.1982; IR (diamond ATR) : 3353 , 2956 (m), 2925 (m), 2869 (m), 1722 (m), 1639 (w), 1446 (m), 1376 (s), 1319 (m), 1258 (w), 1180 (m), 1081 (s), 955 (m), 910 (w), 837 (m), 802 (w) 614 (w), 539 (w) cm−1.I = 1493 ); MS (EI(−)-7-epi-α-Eudesmol (4): GC (HP 5): I = 1661 (literature (HP 5): I = 1662 + calcd for C15H26O+, 204.1873, found. 204.1870; IR (diamond ATR) : 3388 , 2966 (s), 2910 (s), 2850 (s), 1662 (w), 1440 (s), 1376 (w), 1281 (m), 1260 (m), 1220 (s), 1139 (m), 1101 (m), 1021 (s), 937 (w), 873 (m), 841 (s) 798 (w), 755 (w), 707 (w), 649 (w), 568 (w) cm−1.(−)-γ-Cadinene (5): GC (HP 5): I = 1514 (literature (HP 5): I = 1513 + calcd for C15H24+, 204.1873; found, 204.1865; IR (diamond ATR) : 2925 (s), 2850 (s), 1525 (m), 1458 (s), 1366 (s), 1302 (s), 1192 (w), 1087 (s), 1030 (s), 963 (m), 921 (w), 823 (s), 691 (m) 608 (s) cm−1.I = 1513 ); MS (EI(−)-(E)-β-Caryophyllene (6): GC (HP 5): I = 1429 (literature (HP 5): I = 1428 + calcd for C15H24+, 204.1873; found, 204.1868; IR (diamond ATR) : 3066 (w), 2924 (s), 2856 (s), 1670 (m), 1631 (m), 1449 (s), 1382 (s), 1367 (s), 1276 (m), 1257 (w), 1227 (w), 1182 (m), 1106 (w), 1067 (w); 1019 (w), 936 (m), 885 (s), 842 (m), 813 (m), 764 (w) 742 (s), 641 (w), 544 (m) cm−1.I = 1428 ); MS (EIE. coli BL 21 transformants was inoculated from an overnight preculture. The cells were grown to an OD600 = 0.4 before IPTG (0.4 mM) was added. The cultures were incubated at 18 °C and 160 rpm overnight. E. coli cells were harvested by centrifugation at 4 °C and 8000 rpm for 10 min. Protein purification was performed by Ni2+-NTA affinity chromatography with Ni2+-NTA superflow (Novagen) as reported above. For the incubation experiment in 2H2O with 7-epi-α-eudesmol synthase and (6-13C)FPP, the soluble enzyme fraction was washed with binding buffer  and then eluted twice with 10 mL elution buffer . Each pure protein fraction from 0.5 L 2YT liquid culture was concentrated with a Vivaspin20 concentration tube  for 0.5 to 1.5 h at 6000 rpm to 2 mL enzyme fraction. Incubation experiments were performed with the pure protein (2 mL) and the 13C-labeled substrate (0.8 mg to 3.0 mg in 2 mL 2H2O or H2O) at 28 °C overnight. The reaction mixture was extracted with 2 × 0.4 mL (2H6)benzene and directly measured by NMR.For each incubation experiment a 0.5 L 2YT liquid culture (containing kanamycin (50 mg/L)) of File 11–5.Gas chromatograms of extracts from enzyme reactions and NMR spectra of compounds


GNB3) gene has been implicated in obesity risk; however, the molecular mechanism of GNB3-related disease is unknown. GNB3 duplication is responsible for a syndromic form of childhood obesity, and an activating DNA sequence variant (C825T) in GNB3 is also associated with obesity. To test the hypothesis that GNB3 overexpression causes obesity, we created bacterial artificial chromosome (BAC) transgenic mice that carry an extra copy of the human GNB3 risk allele. Here we show that GNB3-T/+ mice have increased adiposity, but not greater food intake or a defect in satiety. GNB3-T/+ mice have elevated fasting plasma glucose, insulin, and C-peptide, as well as glucose intolerance, indicating type 2 diabetes. Fasting plasma leptin, triglycerides, cholesterol and phospholipids are elevated, suggesting metabolic syndrome. Based on a battery of behavioral tests, GNB3-T/+ mice did not exhibit anxiety- or depressive-like phenotypes. GNB3-T/+ and wild-type animals have similar activity levels and heat production; however, GNB3-T/+ mice exhibit dysregulation of acute thermogenesis. Finally, Ucp1 expression is significantly lower in white adipose tissue (WAT) in GNB3-T/+ mice, suggestive of WAT remodeling that could lead to impaired cellular thermogenesis. Taken together, our study provides the first functional link between GNB3 and obesity, and presents insight into novel pathways that could be applied to combat obesity and type 2 diabetes.The G-protein beta subunit 3 ( LEP) as the first obesity gene [delta]Ct method using Inguinal WAT (iWAT) and BAT were dissected, weighed and immediately homogenized on ice in CelLytic MT Mammalian Tissue Lysis/Extraction Reagent  with protease inhibitor cocktail. Total protein concentration was determined using Pierce BCA Protein Assay Kit .Citrate synthase activity for iWAT and BAT extracts were measured using the Citrate Synthase Assay Kit  following the manufacturer’s instructions.t-test was used to compare two groups, and one-way ANOVA was used to compare more than two groups. Comparisons with p-values <0.05 were considered significant.Statistical analyses were performed using GraphPad Prism version 6 for Mac . Data are presented as mean ± SD (or SEM where indicated). Unpaired Student’s GNB3-T/+. To determine the expression levels of human GNB3 and endogenous Gnb3 we performed quantitative RT-PCR in RNA from whole brain, olfactory bulb, hypothalamus, and cerebellum of 5-week-old GNB3-T/+ and WT mice. As expected, we did not detect human GNB3 in WT mice. Notably, human GNB3 expression was much greater than endogenous Gnb3 in whole brain, olfactory bulb, hypothalamus and cerebellum of GNB3-T/+ mice as calculated by delta cycle threshold values , and 50-fold greater than endogenous Gnb3 in iWAT and BAT  were not significantly different in  WT mice . At 20 wther sex . Inflamm obesity . Growth ale mice . It is iGNB3-T/+ mice could be due to greater food intake, lack of activity, or a metabolic defect in energy expenditure. To investigate these possibilities, we measured food consumption at ages 5, 10, 15, 20 and 25 weeks  in male mice. At 5 weeks old, VO2, heat and RER measurements are similar in GNB3-T/+ and WT mice , this difference was not significant.In our transgenic model, GNB3-T/+ mice, indicated by elevated fasting plasma glucose levels and GTT response. Though GNB3-T/+ mice did not have glucose intolerance prior to obesity, 5-week old female GNB3-T/+ mice had elevated fasting blood glucose, and both female and male GNB3-T/+ mice had elevated fasting plasma insulin. This could indicate the beginning stage of impaired glucose metabolism prior to obesity. However, GNB3-T/+ mice do not respond to the ITT like other mouse models of type 2 diabetes, indicating a milder phenotype.Glucose intolerance and type 2 diabetes are apparent at 20 weeks in GNB3-T/+ mice, consistent with lower circulating GH in obese humans [GNB3-T/+ mice at 20 weeks. Thyroid hormones control multiple physiological systems and have an important role in regulating basal metabolic rate, lipolysis, as well as the differentiation process in the adipose tissue [GNB3-T/+ mice at 20 weeks, which could indicate hypogonadotropic hypogonadism [GH was reduced in obese e humans . Anothere tissue . FSH, LHgonadism .GNB3-T/+ mice could be due to increased calorie intake, reduced activity, or a defect in metabolism that results in lower energy expenditure. Our results from food intake measurements and levels of fasting ghrelin, PYY, and amylin hormones revealed that hyperphagia or a satiety defect are not responsible. Novelty suppressed feeding tests also show no significant difference in the amount of sucrose eaten, latency to feed, or total feeding time between the GNB3-T/+ and WT mice, indicating that anxiety-like feeding behaviors are not involved in GNB3-related obesity. GNB3-T/+ and WT mice do not have a statistically significant difference in locomotor activity or oxygen consumption. However, it is possible that subtle differences in locomotion and/or oxygen consumption could contribute to increased adiposity. Future energy expenditure experiments conducted at thermoneutrality and/or brown fat induction experiments could shed light on the effects of GNB3 overexpression. Further, behavioral assessments indicate that there are no substantial anxiety- or depressive-like phenotypes in GNB3-T/+ mice, and that these affective phenotypes are unlikely to add to the relationship between GNB3 overexpression and obesity.Obesity is caused by an energy imbalance between calories consumed and calories expended. The increased adiposity in GNB3-T/+ mice. Ucp1 in mitochondria dissipates chemical energy in the form of heat, mainly in BAT, through a process called nonshivering thermogenesis [+ cells [+ adipose tissue which has a gene expression pattern more similar to beige cells than to classic brown cells in the mouse [GNB3 overexpression alters the gene expression profiles of both BAT and WAT, GNB3 appears to have the greatest effect in beige-cell containing iWAT.Ucp1 expression in adipose tissues provides a clue to the underlying defect in ogenesis . Recentlogenesis . In addi[+ cells –48 known[+ cells  or brown[+ cells . Beige c[+ cells  and deve[+ cells . Adult hhe mouse . Though GNB3-T/+ mice has lower expression of beige and brown adipocyte markers. Additionally, GNB3-T/+ mice showed markedly worsened beige adipocyte function in subcutaneous fat pads as indicated by lower levels of Ucp1 in iWAT. Subcutaneous adipose tissue in GNB3-T/+ mice acquired properties of visceral fat indicated by elevated expression of adipogenic markers, Pparg and adiponectin. Overall, GNB3 overexpression stimulates a conversion of subcutaneous WAT, particularly in the inguinal depot, into a less UCP1+ and a less beige but whiter tissue. We show that this white-like remodeling of iWAT and loss of brown and beige properties in BAT is accompanied by increased adiposity in mice fed normal chow. Together, these data implicate GNB3 overexpression in impaired WAT and BAT, and for the first time provide a functional link between GNB3 and obesity pathogenesis. However, the specific causes of GNB3-related obesity remain to be determined. Future studies of GNB3 overexpression are needed to dissect the molecular mechanisms by which GNB3 alters adipose tissue metabolism, signaling, and energy expenditure.BAT in S1 Fign = 8–11 mice per group in A, C, and D; n = 1–8 mice per group in B; n = 14–22 mice per group in E, F. Data are ± SD. No significant difference between GNB3-T/+ and WT of same sex by unpaired Student’s t-test.A. gWAT weight/body weight (gWAT%), B. iWAT weight/body weight (iWAT%), C. BAT weight/body weight (BAT%), D. liver weight/body weight (liver%), E. DXA lean mass and F. DXA fat mass of 5-week-old mice. (TIF)Click here for additional data file.S2 Fign = 8–11 mice per group in A, n = 7 mice per group in B-D, n = 9–13 mice per group in E-H, n = 10–14 mice per group in I, n = 11–15 mice per group in J. Data are ± SD. *P < 0.05, **P < 0.01 vs. WT of same sex by unpaired Student’s t-test.A. Fasting blood glucose, B. fasting plasma insulin, C. C-peptide, D. leptin, E. triglycerides, F. cholesterol, G. phospholipids and H. non-esterified fatty acids. I. GTT and AUC of female and J. male mice. All mice are 5 weeks old. (TIF)Click here for additional data file.S3 FigGNB3-T/+ mice have elevated fasting plasma resistin during obesity.Male  A,B. Fasting plasma glucagon; C,D. resistin; E,F. GIP; G,H. IL-6; I.J., TNF-a; and K,L. MCP-1 in 5-week and 20-week-old mice, respectively. n = 7 mice per group in A, C, E, G, I, K; n = 10–13 mice per group in B, n = 11–20 mice per group in D, F, H, J, L. Data are ± SD. *P < 0.05 vs. WT of same sex by unpaired Student’s t-test.(TIF)Click here for additional data file.S4 FigGNB3-T/+ male mice have lower GH, TSH, FSH, LH and prolactin during obesity. A,B. Fasting plasma growth hormone (GH); C,D. thyroid-stimulating hormone (TSH); E,F. follicle-stimulating hormone (FSH); G,H. luteinizing hormone (LH); I,J. prolactin; K,L. adrenocorticotropic hormone (ACTH) levels of 5 week and 20 week old mice, respectively. n = 7 mice per group in A, C, E, G, I, K; n = 11–20 mice per group in B, D, F, H, J, L. Data are ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 vs. WT of same sex by unpaired Student’s t-test.(TIF)Click here for additional data file.S5 Fign = 7 mice per group in A-C, n = 19–21 mice per group in D-F. Data are ± SD in A-C, and ± SEM in D-F. *P < 0.05, **P < 0.01, ***P < 0.001 vs. WT of same sex by unpaired Student’s t-test.A. Fasting plasma ghrelin, B. PYY and C. amylin. D. Total movement in X-plane, E. horizontal ambulatory activity, and F. vertical activity averaged over a 72-hour period. All mice are 5 weeks old. (TIF)Click here for additional data file.S6 Fig2, B. heat produced normalized over body weight, and C. RER (VCO2/VO2) averaged over a 72-hour period. D. Acute cold stress at 4°C. All mice are 5 weeks old. n = 19–21 mice per group in A-C, n = 5–8 mice per group in D. Data are ± SEM in A-C; and ± SD in D-F. ****P < 0.0001 vs. WT of same sex by unpaired Student’s t-test.A. VO(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 TableGNB3-T/+ mice and WT littermates were subjected to behavioral tests in order to evaluate anxiety/depressive-like behaviors and learning and memory.(DOCX)Click here for additional data file.


Nature Communications7: Article number: 11929; DOI: 10.1038/ncomms11929 (2016); Published 06222016; Updated 03172017In this Article, residues D128 and D132 of ASC are consistently referred to incorrectly as D130 and D134, respectively. These errors appear in the Results, Methods, Fig. 2, Fig. 3 and Supplementary Fig. 1.


Enterobacter cloacae is a major nosocomial pathogen causing bloodstream infections. We retrospectively conducted a study to assess antimicrobial susceptibility and phylogenetic relationships of E. cloacae bloodstream isolates in two tertiary university-affiliated hospitals in Shanghai, in order to facilitate managements of E. cloacae bloodstream infections and highlight some unknowns for future prevention.E. cloacae bloodstream isolates were consecutively collected from 2013 to 2016. Antimicrobial susceptibility was determined by disk diffusion. PCR was performed to detect extended-spectrum β-lactamase (ESBL), carbapenemase and colistin resistance (MCR-1) gene. Plasmid-mediated AmpC β-lactamase (pAmpC) genes were detected using a multiplex PCR assay targeting MIR/ACT gene  and other plasmid-mediated genes, including DHA, MOX, CMY, ACC, and FOX. eBURST was applied to analyze multi-locus sequence typing (MLST).Fifty-three non-duplicate E. cloacae isolates, 8(15.1%) were ESBL producers, 3(5.7%) were carbapenemase producers and 18(34.0%) were pAmpC producers. ESBL producers bear significantly higher resistance to cefotaxime (100.0%), ceftazidime (100.0%), aztreonam (100.0%), piperacillin (87.5%), tetracycline (75.0%), and trimethoprim-sulfamethoxazole (62.5%) than non-producers (p<0.05). PAmpC- and non-producers both presented low resistance rates (<40%) to all antibiotics (p>0.05). SHV  and MIR/ACT  predominated in ESBL and pAmpC producers respectively. Moreover, 2 isolates co-carried TEM-1, SHV-12, IMP-26 and DHA-1. MLST analysis distinguished the 53 isolates into 51 STs and only ST414 and ST520 were assigned two isolates of each (2/53).The rates of resistance to all tested antibiotics were <40%. Among 53 E. cloacae bloodstream isolates in the two hospitals. Multiclonality disclosed no evidence on spread of these isolates in Shanghai. The simultaneous presence of ESBL, carbapenemase and pAmpC detected in 2 isolates was firstly reported in Shanghai, which necessitated active ongoing surveillances and consistent prevention and control of E. cloacae.The antimicrobial resistance rates were low among 53  Enterobacter cloacae is an important emerging pathogen, causing various nosocomial infections, including respiratory infections, bloodstream infections (BSIs) and surgical site infections ). Escherichia coli ATCC 25922 was used for quality control.Antimicrobial susceptibility of the  by CLSI . The ant Edition). For tig Edition); for polblaTEM, blaSHV, blaCTX-M , blaOXA , blaVEB, blaGES, and blaPER), carbapenemase genes  and plasmid-mediated colistin resistance gene (mcr-1), using primers previously described [http://blast.ncbi.nlm.nih.gov/BLAST).Polymerase chain reaction (PCR) was performed to confirm the existence of ESBL genes  as described by Miyoshi-Akiyama et al.[http://pubmlst.org/ecloacae/). STs not found in the database were submitted. The eBURST version 3.0 was used to analyze the clustering of related STs. The eBURST algorithm can group strains according to their allelic profiles by employing a user-specified group definition, as well as drawing a rough sketch to show the genetic relationship[A multilocus sequence typing (MLST) scheme was used to assign ma et al.. The comationship. In thisP value of <0.05.SAS 8.2  was used for statistical analysis. Continuous variables were presented as median and interquartile range. The chi-square or Fisher’s exact test was used to compare the disparity between different groups for categorical variables. Differences were considered statistically significant at a two-tailed The age of the 53 patients ranged from 23 to 86 years . Male patients (37/53) were more than females (16/53). Near half of the episodes were derived from the surgery (22/53), and where most patients suffered from malignant tumor (10/22) .E. cloacae isolates, 8(15.1%) were ESBL producers, 3(5.7%) were carbapenemase producers and 18(34.0%) were pAmpC producers (P<0.05). Of note, pAmpC- and non-producers both presented low resistance rates (<40%) to all antibiotics, which did not differ significantly in the two groups (p>0.05) (As summarized in roducers . The hig(p>0.05) .E. cloacae isolates were positive for pAmpC genes, of which 15 (83.3%) isolates were detected with MIR/ACT gene and the other 3 (16.7%) were detected with DHA-1 gene , blaVEB, blaGES, blaPER, blaVIM, blaKPC, blaOXA-48 and blamcr-1 were detected.Of the 8 ESBL producers, SHV enzymes  predominated with all identified as SHV-12, and CTX-M enzymes  followed, including CTX-M-15 and CTX-M-65  . A totalA-1 gene . Of the A-1 gene . Besideshttps://pubmlst.org/bigsdb?db=pubmlst_ecloacae_seqdef). It was indicated in MLST analysis distinguished 51 different STs, clustered into 3 non-overlapping clonal complexes (CCs) and 43 singletons . Only STE. cloacae was frequently implicated in nosocomial infections[E. cloacae isolates[E. cloacae worsening clinical outcome and prolonging hospitalization duration have been alarmingly increasing since 21st century[E. cloacae bloodstream isolates harboring resistant genes were mostly associated with elderly patients from the surgery or Intensive Care Unit  produced ESBL, which was higher than that in north-eastern USA (10.1%)[Enterobacter spp. collected from all sites[E. cloacae [E. cloacae isolates due to various modes of transmission.Of all 15.1% 8/5 producedall sites. Unlike all sites, SHV wasall sites, and simall sites. SHV-12, cloacae . What’s  cloacae , 26, whiE. cloacae was once reported in Taiwan[PAmpC can preferentially hydrolyze all β-lactams except fourth-generation cephalosporins and carbapenems and may serve as the reservoir for the emergence of antibiotic resistance . The higin Taiwan, but firPseudomonas aeruginosa isolate in Singapore [E. cloacae isolates, but interestingly, the two IMP-producers in our study were both detected as IMP-26. It was still noted that IMP-producing E. cloacae had caused several outbreaks or possible spread in other countries [E. cloacae. NDM-1 was the dominant carbapenemase of carbapenems-resistant E. cloacae in Henan China, and in contrast to VIM-1 the most prevalent in Spain and other southern Europe [Carbapenemase spread has been increasingly reported worldwide over the last decade. We founingapore  and few ountries , 33, 34,n Europe , 36; whiE. cloacae has once denoted the association of distinct clonal groups with genetic lineages of higher prevalence and/or wider geographical spread [E. cloacae bloodstream isolates did not evolve from a unique ancestral background with 51 STs distinguished from 53 isolates and 8 isolates (15.1%) assigned to 3 CCs Click here for additional data file.


VOLUME 104104(3) July, page 242http://dx.doi.org/10.3163/1536-5050.104.3.014.Bramer WM, Giustini D, de Jonge GB, Holland L, Bekhuis T. De-duplication of database search results for systematic reviews in EndNote. J Med Libr Assoc. 2016 Jul;104(3):240–3. DOI: Page 242: Table 1, row C3, should state:Review the top references without page numbers and those with page numbers starting with the number 1 for equivalent author names.Only articles without page numbers or page numbers starting with 1 should be reviewed.


Correction to:The ISME Journal (2017) 11, 932–944; doi:10.1038/ismej.2016.172Updated online 28 February 2017: This article was originally published under a CC BY-NC-SA 4.0 license, but has now been made available under a CC-BY 4.0 license. The PDF and HTML versions of the paper have been modified accordingly.


In the original article, there was a mistake in the published supplementary material. The literature search string as mentioned in the section “Literature Search and Selection Strategy” never appeared in the published material. The search string as applied in Medline appears below. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way.Ovid Technologies, Inc. Email ServiceDatabase: Epub Ahead of Print, In-Process & Other Non-Indexed Citations, Ovid MEDLINE(R) Daily and Ovid MEDLINE(R)<1946 to Present>Search Strategy:1 Hyperparathyroidism, Primary/dt [Drug Therapy] (99)2 Hyperparathyroidism, Primary/(2211)3 Parathyroid adenom*.mp. (4188)4 (Primary adj2 Hyperparathyr*).mp. (9043)5 2 or 3 or 4 (11256)6 exp Diphosphonates/(22931)7 bisphosphonate*.mp. (14211)8 diphosphonat*.mp. (17321)9 bisphosphonic*.mp. (152)10 clodron*.mp. (2326)11 alendron*.mp. (4714)12 etidron*.mp. (3123)13 ibandron*.mp. (1063)14 incadron*.mp. (78)15 medron*.mp. (4405)16 minodron*.mp. (104)17 neridron*.mp. (94)18 olpadron*.mp. (77)19 pamidron*.mp. (2920)20 risedron*.mp. (1775)21 tiludron*.mp. (156)22 zoledron*.mp. (4145)23 cinacalcet.mp. (1071)24 Cinacalcet Hydrochloride/(735)25 "amg 073".mp. (24)26 amg073.mp. (0)27 krn 1493.mp. (3)28 krn1493.mp. (4)29 mimpara.mp. (20)30 parareg.mp. (0)31 regpara.mp. (1)32 sensipar.mp. (30)33 exp Isoflavones/(15928)34 ipriflavone.mp. (268)35 Denosumab/(925)36 denosumab.mp. (1712)37 amg 162.mp. (33)38 amg162.mp. (3)39 amgiva.mp. (0)40 prolia.mp. (32)41 xgeva.mp. (16)42 blosozumab.mp. (14)43 ly 2541546.mp. (0)44 ly2541546.mp. (1)45 romosozumab.mp. (45)46 amg 785.mp. (27)47 amg785.mp. (0)48 cdp 7851.mp. (0)49 cdp7851.mp. (3)50 odanacatib.mp. (156)51 ("mk 0822" or mk0822 or mk822 or mk 822).mp. (5)52 etelcalcetide.mp. (5)53 (amg 416 or amg416).mp. (12)54 (kai 4169 or kai4169).mp. (1)55 (ono 5163 or ono5163).mp. (0)56 telcalcetide.mp. (0)57 velcalcetide.mp. (4)58 or/6-57 (49081)59 5 and 58 (459)60 1 or 59 (491)The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


We report here the first complete genome sequences of genotype GI.3, GI.4, GI.6, GI.7, and GII.7 sapovirus strains, detected from fecal samples of acute gastroenteritis patients. Complete or nearly complete genome sequences of all 18 genotypes of human sapoviruses are now available for phylogenetic analysis and primer design. Genetically diverse sapovirus (SaV) strains have been detected in fecal specimens from patients with acute gastroenteritis –4. We ha–6–de novo assembly of consensus SaV genome sequence were performed first, as described ((AY237420), GII.3 (AY603425), GII.5 (LC190463), and GII.6 (AY646855) SaV genomes in combination with two primers designed in the previously determined VP1-encoding-region sequence  or QIAamp viral RNA minikit (Qiagen). Library preparation from the purified RNA for sequencing on an Illumina MiSeq platform (Illumina) and escribed , 10. Forescribed  using thsequence . FurtherGII.5 LC10463, andsequence 11. The 5GII.5 LC10463, andThe genomes of GI.3 Hu/OH08021/2008/JP, GI.4 Hu/SV/Chiba/000496/2000, GI.6 Hu/SV/Chiba/000764/2000, GI.7 Hu/D1714-B/2008/JPN, and GII.7 Hu/20072248/2008/JP SaV strains consist of 7,442, 7,436, 7,443, 7,452, and 7,462 nt, respectively, excluding the poly(A) tail. All of these SaV genomes were predicted to encode two ORFs, a short 5′ untranslated region (UTR) (12 or 13 nt long) and a 3′-UTR 78 to 112 nt long). The 5′ terminal sequence was conserved as GTG, similarly to those of other SaVs , 11.2 nt longThe new sequence data determined in this study will be useful in designing more broadly reactive primers and probes for human SaV detection PCR, as well as establishment of a nonstructural protein coding region typing scheme like that recently established for norovirus .AB623037, AJ606693, AJ606694, AB522390, and AB630067, respectively.The genome sequences of GI.3 Gu/OH08021/2008/JP, GI.4 Hu/SV/Chiba/000496/2000, GI.6 Hu/SV/Chiba/000764/2000, GI.7 Hu/D1714-B/2008/JPN, and GII.7 Hu/20072248/2008/JP SaV strains have been deposited in GenBank under the accession no.


Nature Communications8: Article number: 15110; DOI: 10.1038/ncomms15110 (2017); Published: 04272017; Updated: 0613201752’ and ‘pSANG10-3F-BG4 (Addgene 55756)52 was transformed into….’ The correct citation is given below.In this Article, the antibody ‘BG4’ is consistently referred to incorrectly as ‘hf2’. These errors appear in the Results and Methods. Additionally, the Article incorrectly cites reference 52 in the sentences ‘A recent study described hf2, an engineered single-chain antibody specific for G4 DNANat. Chem.5, 182–186 (2013).Biffi, G., Tannahill, D., McCafferty, J. & Balasubramanian, S. Quantitative visualization of DNA G-quadruplex structures in human cells.