Correction to: J Immuno Therapy Cancer. (2018) 6 (Suppl 1)https://doi.org/10.1186/s40425-018-0422-yhttps://doi.org/10.1186/s40425-018-0423-xAfter publication of this supplement , 2, it wP128Multiplexed immunofluorescent assay development for study of the PD-1/PD-L1 checkpoint in the tumor immune microenvironment (TIME)Sneha Berry, MS, Nicolas Giraldo, MD PhD, Peter Nguyen, MS, Benjamin Green, BS, Haiying Xu, Aleksandra Ogurtsova, Abha Soni, DO, Farah Succaria, MD, Daphne Wang, MS, Charles Roberts, Julie Stein, MD, Elizabeth Engle, MSc, Drew Pardoll, MD, PhD, Robert Anders, MD, PhD, Tricia Cottrell, MD, PhD, Janis M. Taube, MDJohns Hopkins University School of Medicine, Baltimore, MD, USACorrespondence: Sneha Berry (jtaube1@jhmi.edu)Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P128Janis M. Taube, MD is a contributing author and has therefore been added to the author list in this correction article. Contributing author Sneha Berry, MS is listed three times in the original article; this is no longer the case in this correction article.BackgroundMultispectral immunofluorescent (mIF) staining of formalin-fixed paraffin-embedded (FFPE) tissue allows spatially-resolved quantitative analysis of cell position and protein expression. The design and validation of mIF panels is a challenge. Our goal was to develop a 7-plex assay for characterizing PD-1 and PD-L1 expression, with high sensitivity for multiple markers and minimal bleed-through between fluorescent channels, while avoiding steric hindrance among markers occupying the same cellular compartment.MethodsSingle IF slides were stained for PD-1, PD-L1, CD8, FoxP3, CD163, and a tumor marker (e.g. Sox10/S100 for melanoma) using primary antibodies at manufacturer’s recommended concentrations and visualized with an Opal kit (PerkinElmer). Positive signal was compared to chromogenic IHC (n=3 tonsil specimens). In some instances, the kit’s HRP-polymer was substituted for one that provided greater amplification. Primary antibody titrations were performed, and the concentration with comparable signal to chromogenic IHC that showed the highest IF signal to noise ratio was selected. Using the selected primary antibody concentration, TSA dilution series were performed on n=5 tumor specimens to minimize bleed-through. Finally, the optimized single IF stains were combined into multiplex format, which was again validated to ensure no positivity loss. Images were scanned with the Vectra 3.0 and processed using inForm (Ver 2.3).Resultshigh vs. PD-1low lymphocytes.The percent positive pixels for CD163, CD8, and tumor marker expression by IF were comparable to chromogenic IHC with manufacturer’s recommended protocols (p>0.05). However, PD-1, PD-L1, and FoxP3 showed ~50% loss of signal (p<0.05), which was recovered by replacing the Opal kit's secondary HRP polymer with PowerVision (Leica). Unbalanced fluorescence intensities between 540 to 570 Opal dyes resulted in significant bleed-through and led to false positive pixels. This error was minimized >2 fold (2.5% to 1.1%) by concentrating the 570 dye and ensuring that this dye pair was used to study markers in different cellular compartments (nuclear FoxP3 vs. membrane CD8), so any residual bleed-through could be discounted during image analysis. Using the optimized panel, we are able to reliably identify cell types contributing PD-L1 and PD-1 to the TIME, and even resolve populations of PD-1ConclusionsWe demonstrate successful optimization of a 7-color multiplex panel characterizing the PD-1/PD-L1 axis to provide high quality data sets for whole slide or regional analysis of the TIME. With the use of multiparametric assays such as this, we hope to guide improved approaches to patient selection and potentially identify additional tumor types likely to respond to anti-PD-(L)1 immunotherapy.Ethics ApprovalThe study was approved by Johns Hopkins University Institutional Review Board.P308A Phase 1 study of MEDI5752, a bispecific antibody that preferentially targets PD-1 and CTLA-4 expressing T cells, in patients with advanced solid tumors2, Mark Voskoboynik3, James Kuo4, Yung-Lue Bang5, Hyun-Cheo Chung6, Myung-Ju Ahn7, Sang-We Kim8, Ayesh Perera1, Daniel Freeman1, Ikbel Achour1, Raffaella Faggioni1, Feng Xiao1, Charles Ferte1, Charlotte Lemech4Ben Tran1MedImmune, Gaithersburg, MD, USA; 2Peter MacCallum Cancer Center, Melbourne, Australia; 3Nucleus Network, Melbourne, Australia; 4Scientia Clinical Research, Sydney, Australia; 5Seoul National University Hospital, Seoul, Korea, Republic of; 6Yonsei Cancer Center, Yonsei University, Seoul, Korea, Republic of; 7Samsung Medical Center, Seoul, Korea, Republic of; 8Asan Medical Center, Songpa-Gu, Korea, Republic ofCorrespondence: Charles Ferte (fertec@MedImmune.com) Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P308Jeff Brubaker was not a contributing author and has therefore been removed from the author list in this correction article. The credentials as shown on the original article are no longer listed on this correction article.BackgroundBased on demonstrated clinical activity and manageable safety profiles, checkpoint inhibiting antibodies blocking PD 1, PD-L1, or CTLA-4 have received regulatory approvals for the treatment of various malignancies [1-5]. The combination therapy with anti-PD-1 and anti-CTLA-4 agents is approved by FDA for metastatic melanoma, renal cell carcinoma and microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR) metastatic colorectal cancer, based on improved overall survival versus either agent alone [6-10]. Numerous clinical studies of combination immunotherapy are currently investigating the same combination across a range of solid tumors [11- 15]. Although the efficacy of these drug combinations is dose dependent, the toxicity associated with anti-CTLA-4 agents, in particular, is dose limiting, thereby potentially affecting treatment outcomes with combination therapy.- MEDI5752 is a bispecific humanized IgG1 monoclonal antibody that binds PD-1 and CTLA-4. In contrast to the combination therapy, MEDI5752 exhibits a novel T cell targeting mechanism that could provide a favorable toxicity profile. In addition, we have shown that MEDI5752 can impact cell surface expression of PD-1. Based on these novel mechanisms of action, MEDI5752 may show improved efficacy and safety in comparison to co- administration of conventional anti-PD1/anti-PD-L1 and anti-CTLA-4 antibodies.MethodsThis is a Phase 1, first-time-in-human, multicenter, open-label study in patients with advanced solid tumors. The dose-escalation phase will evaluate approximately six MEDI5752 dose levels to identify a maximum tolerated dose. Dose escalation will be followed by two dose-expansion cohorts in defined setting with patients with advanced or metastatic solid tumor and tested against a control arm. Subjects will remain on treatment until confirmed progressive disease, initiation of alternative cancer therapy, unacceptable toxicity, or other reason for discontinuation. The primary endpoints are safety and efficacy (objective response in the dose-expansion phase). Secondary endpoints include additional efficacy assessment across both phases, pharmacokinetics, and immunogenicity.Trial RegistrationNCT03530397References1. Opdivo Prescribing Information. Princeton, NJ: Bristol-Myers Squibb Company, 2018. [revised 2018 Apr].2. Keytruda Prescribing Information. Whitehouse Station, NJ: Merck Sharp & Dohme Corp, 2018 [revised 2018 Jun].3. Tecentriq Prescribing Information. San Francisco (CA): Genentech, Inc, 2018. [revised 2019 Jul].4. Imfinzi Prescribing Information. Wilmington (DE): AstraZeneca Pharmaceuticals LP, 2018. [revised 2018 Feb].5. Yervoy Prescribing Information. Princeton (NJ): Bristol-Myers Squibb Company, 2018. [revised 2018 Apr].6. Larkin J, Chiarion-Sileni V, Gonzalez R, Grob JJ, Cowey CL, Lao CD, et al. Combined nivolumab and ipilimumab or monotherapy in untreated melanoma. N Engl J Med. 2015;373:23-34.7. Postow MA, Chesney J, Pavlick AC, Robert C, Grossmann K, McDermott D, et al. Nivolumab and ipilimumab versus ipilimumab in untreated melanoma. N Engl J Med. 2015;372:2006-2017.8. Motzer RJ, Tannir NM, McDermott DF, Frontera OA, Melichar B, Choueiri TK, et al. Nivolumab plus ipilimumab versus sunitinib in advanced renalcell carcinoma. N Engl J Med. 2018;378:1277-1290.9. Overman MJ, Lonardi S, Wong KYM, Lenz H-J, Gelsomino F, Aglietta M, et al. Durable clinical benefit with nivolumab plus ipilimumab in DNA mismatch repair–deficient/microsatellite instability–high metastatic colorectal cancer. J Clin Oncol. 2018;36:773-779.https://www.pharmacytimes.com/news/nivolumab-ipilimumabcombo-approved-by-fda-for-msihdmmr-colorectal-cancer. Accessed 18 July, 2018.10. Broderick JM. Nivolumab, ipilimumab combo approved by FDA for MSI-H/dMMR colorectal cancer. Pharmacy Times, 11 July, 2018. Available at: 11. Antonia S, Goldberg SB, Balmanoukian A, Chaft JE, Sanborn RE, Gupta A, et al. Safety and antitumour activity of durvalumab plus tremelimumab in non-small cell lung cancer: a multicentre, phase 1b study. Lancet Oncol. 2016a;17(3):299-308.12. Haddad R, Gillison M, Ferris RL, Harrington K, Monga M, Baudelet C, et al. Double-blind, two-arm, phase 2 study of nivolumab (nivo) in combination with ipilimumab (ipi) versus nivo and ipi-placebo (PBO) as first-line (1L) therapy in patients (patients) with recurrent or metastatic squamous cell carcinoma of the head and neck (R/M SCCHN)—CheckMate 714. Ann Oncol. 2016;27(suppl_6):1017TiP-TiP.13. Kelley RK, Abou-Alfa GK, Bendell JC, Kim T-Y, Borad MJ, Yong W-P, et al. Phase I/II study of durvalumab and tremelimumab in patients with unresectable hepatocellular carcinoma (HCC): Phase I safety and efficacy analyses. J Clin Oncol. 2017;35(15_suppl):4073.14. Janjigian YY, Ott PA, Calvo E, Kim JW, Ascierto PA, Sharma P, et al. Nivolumab ± ipilimumab in patients with advanced (adv)/metastatic chemotherapy-refractory (CTx-R) gastric (G), esophageal (E), or gastroesophageal junction (GEJ) cancer: CheckMate 032 study. J Clin Oncol. 2017;35(15_suppl):4014.15. Escudier B, Tannir NM, McDermott DF, Frontera OA, Melichar B, Plimack ER, et al. LBA5CheckMate 214: Efficacy and safety of nivolumab + ipilimumab (N+I) v sunitinib (S) for treatment-naïve advanced or metastatic renal cell carcinoma (mRCC), including IMDC risk and PD-L1 expression subgroups. Ann Oncol. 2017;28(suppl_5):mdx440.029- mdx440.029.P309Phase I dose-finding study of MIW815 (ADU-S100), an intratumoral STING agonist, in patients with advanced solid tumors or lymphomas2, Theresa Werner3, Stephen Hodi4, Wells Messersmith, MD5, Nancy Lewis6, Craig Talluto7, Mirek Dostalek6, Aiyang Tao6, Sarah McWhirter8, Damian Trujillo8, Jason Luke, MD, FACP9Funda Meric-Bernstam, MD2MD Anderson Cancer Center, Houston, TX, USA; 3University of Utah, Salt Lake City, UT, USA; 4Dana-Faber Cancer Institute, Boston, MA, USA; 5University of Colorado Cancer Center, Aurora, CO, USA; 6Novartis Pharmaceuticals Corporation, East Hanover, NJ, USA; 7Novartis Institutes for BioMedical Resea, Cambridge, MA, USA; 8Aduro Biotech Inc, Berkeley, CA, USA; 9The University of Chicago Medicine, Chicago, IL, USACorrespondence: Funda Meric-Bernstam (fmeric@mdanderson.org)Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P309Janis Callister was not a contributing author and has therefore been removed from the author list in this correction article. The redundant affiliation Articulate Science as shown on the original article is no longer listed on this correction article.BackgroundMIW815 (ADU-S100) is a novel synthetic cyclic dinucleotide that can activate human STING (STimulator of INterferon Genes) in antigenpresenting cells. In preclinical models, STING pathway activation can induce tumor antigen-specific T-cell priming within the tumor microenvironment, leading to antitumor immunity and tumor destruction.MethodsEligible patients  include those with advanced/metastatic solid tumors or lymphomas with progressive disease despite standard of care or for whom there is no standard treatment.MIW815 (ADU-S100) is administered by weekly intratumoral injections (3 weeks on/1 week off) at escalating doses (starting dose: 50μg) in 28-day cycles. Primary objectives are to characterize safety and tolerability and to identify a recommended dose for future studies.Secondary objectives include preliminary efficacy, pharmacokinetics (PK), and pharmacodynamics (PD). The study is currently in dose escalation.ResultsAs of June 15, 2018, 41 heavily pretreated patients  with various solid tumors or lymphomas were enrolled. Thirty-five patients have discontinued from the study for the following reasons: disease progression (n=26), physician/patient decision (n=8), and death (n=1); 6 patients continue to receive treatment. No dose-limiting toxicities (DLTs) were reported during the first cycle at any dose level. The most common (≥10% of patients) treatment-related AEs (TRAEs) were pyrexia , injection site pain , and headache . Grade 3/4 TRAEs included increased lipase  and elevated amylase, tumor pain, dyspnea, respiratory failure, and injection site reaction . Systemic MIW815 (ADU- S100) exposure increased with dose. On-treatment tumor biopsies showed increases in CD8 T cells infiltrating the injected tumors in a subset of patients. Preliminary antitumor activity, PK analysis, and PD data from injected lesions, noninjection lesions, and peripheral blood, will be presented.ConclusionsIntratumoral injection of MIW815 (ADU-S100) was well tolerated in doses tested thus far in patients with advanced solid tumors and lymphoma, with no DLTs reported to date. Trials evaluating combinations of MIW815 (ADU-S100) with anti-PD1 or anti-CTLA4 antibodies are ongoing.Ethics ApprovalThis study was approved by an independent ethics committee or institutional review board at each site.P383Combination therapy with M7824 (MSB0011359C) and NHSmuIL12 enhances antitumor efficacy in preclinical cancer models2, Bo Marelli2, Jin Qi2, Guozhong Qin2, Huakui Yu2, Molly Jenkins2, Kin-Ming Lo2, Joern-Peter Halle2, Yan Lan, MD2Chunxiao Xu, PhD2EMD Serono Research and Development, Belmont,MA, USACorrespondence: Yan Lan (yan.lan@emdserono.com)Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P383Colleen Stanton was not a contributing author and has therefore been removed from the author list in this correction article. The redundant affiliation Nucleus Global as shown on the original article is no longer listed on this correction article.BackgroundPD-1/PD-L1 pathway inhibition is a clinically validated approach in cancer therapy. However, most patients do not respond to the monotherapy due to multiple immunosuppressive mechanisms. Combining anti-PD-1/ PD-L1 with other immunotherapeutic agents targeting additional immunomodulatory pathways in the tumor microenvironment (TME) is one strategy to overcome resistance and improve response rates. M7824 is an innovative first-in-class bifunctional fusion protein composed of two extracellular domains of TGF-β receptor II (a TGF-β “trap”) fused to a human anti-PD-L1 IgG1 monoclonal antibody. Through simultaneous blockade of the PD-L1 and TGF-β pathways, M7824 demonstrated enhanced anti-tumor activity in preclinical models [1]. NHS-IL12, and the surrogate NHS- muIL12, are immunocytokines designed to target tumor necrotic regions to deliver IL-12 into the TME, where they can activate NK cells and CD8+ T cells to increase their cytotoxic functions. The surrogate NHSmuIL12 has demonstrated antitumor efficacy in preclinical models [2].This study is designed to investigate whether M7824 treatment may further benefit from combination therapy with NHS-muIL12.MethodsMice bearing MC38, EMT-6, or 4T1 tumors were treated with M7824, NHS-muIL12, or combination therapy. Tumor growth and survival were assessed in each model, and tumor recurrence following remission and rechallenge was evaluated in the EMT-6 model. Immune cell populations in the spleens and tumors were evaluated by flow cytometry and the frequency of tumor antigen-reactive IFNγ-producing CD8+ T cells was evaluated by an ELISpot assay in the MC38 model.ResultsCombination of M7824 and NHS-muIL12 enhanced antitumor activity and extended the survival relative to either monotherapy in preclinical tumor models. Combination therapy also enhanced the proliferation, infiltration, and cytotoxicity of CD8+ T cells relative to monotherapies. In addition, the combination therapy increased the frequency of tumor antigenreactive T cells and induced the generation of tumor-specific immune memory, as demonstrated by protection against tumor rechallenge.ConclusionsThese data demonstrate that combination therapy with M7824 and NHS-muIL12 improved anti-tumor efficacy in multiple preclinical tumor models and suggest that combining these therapies may be a promising therapeutic strategy for patients with solid tumors.References1. Lan Y, et al. Enhanced preclinical antitumor activity of M7824, a bifunctional fusion protein simultaneously targeting PD-L1 and TGF-β. Sci Trans Med. 2018;10(424).2. Fallon J, et al. The immunocytokine NHS-IL12 as a potential cancer therapeutic. Oncotarget 2014;5(7):1869-84.P391A phase 1b/2 trial of lenvatinib in combination with pembrolizumab in patients with advanced melanoma2, Nicholas Vogelzang, MD3, Allen Cohn3, Daniel Stepan4, Robert Shumaker, PhD,4, Corina Dutcus4, Matthew Guo4, Emmett Schmidt, MD PhD5, Drew Rasco6Matthew Taylor, MD2Oregon Health and Science University, Portland, OR, USA; 3McKesson Specialty Health, Las Vegas, NV, USA; 4Eisai Inc., Woodcliff Lake, NJ, USA; 5Merck & Co., Inc., Kenilworth, NJ, USA; 6South Texas Accelerated Research Therape, San Antonio, TX, USACorrespondence: Matthew Taylor (taylmatt@ohsu.edu) Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P391The redundant affiliation 1. Oxford PharmaGenesis, Oxford, UK as shown on the original article is no longer listed on this correction article.BackgroundLenvatinib is a multikinase inhibitor of VEGFR 1−3, FGFR 1−4, PDGFRα, RET, and KIT. Pembrolizumab, an anti-PD-1 antibody, is approved for the first-line treatment of patients with advanced melanoma, with objective response rates (ORR) of 21–34% . Preclinical studies indicate that lenvatinib decreases the population of tumorassociated macrophages, increases CD8+ T cell infiltration, and augments the activity of PD-1 inhibitors; therefore, lenvatinib is a rational combination partner for pembrolizumab . We report interim results of an ongoing phase 1b/2 trial evaluating lenvatinib in combination with pembrolizumab in patients with solid tumors, focusing on the advanced melanoma cohort.MethodsIn this multicenter, open-label study (NCT02501096), patients with measurable, confirmed, metastatic melanoma and ECOG performance status ≤1 received lenvatinib  + pembrolizumab . Patients were not preselected based on PDL1 status. Tumor assessments were performed by study investigators using immune-related RECIST (irRECIST). The phase 2 primary end point was ORR at 24 weeks (ORRWK24). Secondary end points included ORR, progression-free survival (PFS), and duration of response (DOR).ResultsAt the data cutoff of March 1, 2018, 21 patients were enrolled: 14 (67%) patients were PD-L1(+), 4 (19%) were PD-L1(-), 3 (14%) were not tested; and 38% of patients had ≥1 prior anticancer therapy. The primary end point of ORRWK24 was 47.6% . Additional efficacy outcomes are summarized in the table (Table 1). All patients experienced ≥1 treatment-related adverse event (TRAE). Grade 3 and 4 TRAEs occurred in 13 (62%) and 1  patients respectively. There were no fatal TRAEs. The most common any-grade TRAEs were fatigue (52%), decreased appetite (48%), diarrhea (48%), hypertension (48%), dysphonia (43%), and nausea (43%). Dose reduction and interruption due to TRAEs occurred in 13 (62%) and 10 (48%) patients, respectively.ConclusionsThe lenvatinib and pembrolizumab combination regimen was welltolerated and demonstrated encouraging clinical activity. The combination may potentially improve upon the antitumor activity of antiPD-1 monotherapies, supporting further evaluation of this regimen in patients with advanced melanoma.Trial RegistrationNCT02501096References1. Ribas A, et al. Pembrolizumab versus investigator-choice chemotherapy for ipilimumab-refractory melanoma (KEYNOTE-002): a randomised, controlled, phase 2 trial. Lancet Oncology. 2015;16(8):908-18.2. Robert C, et al. Pembrolizumab versus ipilimumab in advanced melanoma. N Engl J Med. 2015;372(26):2521- 2532.3. Kato Y et al. Effects of lenvatinib on tumor-associated macrophages enhance antitumor activity of PD-1 signal inhibitors. Mol Cancer Ther. 2015;14 (12Suppl2):A92.4. Kato Y. Upregulation of memory T cell population and enhancement of Th1 response by lenvatinib potentiate antitumor activity of PD-1 signaling blockade. Cancer Res. 2017;77 (13 Suppl):4614. Ethics Approval This study was approved by all relevant institutional review boards.Table 1 (abstract P391). See text for description.P392A phase 1b/2 trial of lenvatinib in combination with pembrolizumab in patients with non-small cell lung cancer2, Nicholas Vogelzang, MD3, Christopher Di Simone3, Sharad Jain, MD3, Donald Richards3, Carlos Encarnacion3, Drew Rasco4, Robert Shumaker, PhD5, Corina Dutcus5, Daniel Stepan5, Matthew Guo5, Emmett Schmidt, MD PhD6, Matthew Taylor, MD7Marcia Brose2Abramson Cancer Center of the University, Philadelphia, PA, USA; 3McKesson Specialty Health, Las Vegas, NV,USA; 4South Texas Accelerated Research Therape, San Antonio, TX, USA; 5Eisai Inc., Woodcliff Lake, NJ, USA; 6Merck & Co., Inc., Kenilworth, NJ, USA; 7Oregon Health and Science University, Portland, OR, USACorrespondence: Marcia Brose (Brosem@mail.med.upenn.edu) Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P392The redundant affiliation 1. Oxford PharmaGenesis, Oxford, UK as shown on the original article is no longer listed on this correction article.BackgroundLenvatinib is a multikinase inhibitor of VEGFR 1−3, FGFR 1−4, PDGFRα, RET, and KIT. Pembrolizumab, an anti-PD-1 antibody, is approved as a monotherapy for previously treated patients with metastatic PD-L1–positive (tumor proportion score [TPS] ≥1%) non-small cell lung cancer (NSCLC), with an objective response rate (ORR) of 18% [1]. We report interim results of an ongoing phase 1b/2 trial evaluating lenvatinib in combination with pembrolizumab in patient with solid tumors, focusing on the metastatic NSCLC cohort.MethodsIn this multicenter, open-label study (NCT02501096), patients with measurable, confirmed metastatic NSCLC and ECOG performance status ≤1 received lenvatinib  and pembrolizumab . In the phase 2 portion, patients must have had ≤2 prior lines of systemic therapy; there was no limit for phase 1b. Patients were not preselected based on PD-L1 status. Tumor assessments were performed by study investigators using immune-related RECIST (irRECIST). The phase 2 primary end point was ORR at 24 weeks (ORRWK24). Secondary end points included ORR, progressionfree survival (PFS), and duration of response (DOR).ResultsAt the data cutoff of March 1, 2018, 21 patients were enrolled. 9 (43%) Patients were PD-L1(+) (TPS ≥1%); 5 (24%) were PD-L1(-); 7 (33%) were not tested. 3 (14%) Patients were treatment-naïve; 7 (33%), 10 (48%), and 1 (5%) patients had 1, 2, and ≥3 prior lines of systemic therapy, respectively. The primary end point of ORRWK24 was 33.3% . Additional efficacy outcomes are summarized in the table (Table 1). Grade 3 and 4 treatment-related adverse events (TRAEs) occurred in 10 (48%) and 1  patients, respectively. There was 1 fatal TRAE . The most common grade 3 TRAEs were hypertension (24%), fatigue (14%), diarrhea (14%), proteinuria (10%), and arthralgia (10%).ConclusionsThe combination of lenvatinib and pembrolizumab showed promising clinical activity with a manageable safety profile in previously treated patients with metastatic NSCLC who were not preselected for PD-L1 status. Further study is warranted.Trial RegistrationNCT02501096ReferencesHerbst RS et al. Pembrolizumab versus docetaxel for previously treated, PD-L1-positive, advanced non-small-cell lung cancer (KEYNOTE-010): a randomised controlled trial. Lancet. 2016;387(10027):1540-50.Ethics ApprovalThis study was approved by all relevant institutional review boards.Table 1 (abstract P392). See text for description.P393A phase 1b/2 trial of lenvatinib in combination with pembrolizumab in patients with urothelial cancer2, Carlos Encarnacion2, Allen Cohn2, Christopher Di Simone2, Drew Rasco3, Donald Richards2, Matthew Taylor, MD4, Corina Dutcus5, Daniel Stepan5, Robert Shumaker, PhD5, Matthew Guo5, Emmett Schmidt, MD PhD6, James Mier, MD7Nicholas Vogelzang, MD2McKesson Specialty Health, Las Vegas, NV, USA; 3South Texas Accelerated Research Therape, San Antonio, TX, USA; 4Oregon Health and Science University, Portland, OR, USA; 5Eisai Inc., Woodcliff Lake, NJ, USA; 6Merck & Co., Inc., Kenilworth, NJ, USA; 7Beth Israel Deaconess Medical Center, Boston, MA, USACorrespondence: Nicholas Vogelzang (nicholas.vogelzang@usoncology.com) Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P393The redundant affiliation 1. Oxford PharmaGenesis, Oxford, UK as shown on the original article is no longer listed on this correction article.BackgroundPembrolizumab, an anti-PD-1 antibody, is approved in the secondline setting for patients (objective response rate [ORR] 21%) with advanced/metastatic urothelial cancer and in the first-line setting for patients who are ineligible for cisplatin with combined positive score ≥10 or ineligible for platinum-based chemotherapy, with ORR  [1–3]. However, there is still an unmet need for effective therapeutic options for advanced urothelial cancer. Lenvatinib is a multikinase inhibitor of VEGFR 1-3, FGFR 1-3, PDGFRα, RET and KIT. Tyrosine kinase inhibitors, such as lenvatinib, have demonstrated activity in urothelial cancer and may reverse the immunosuppressive environment that leads to immuno-oncology (IO) therapy failure. Here we present a phase 1b/2 trial to determine the safety and efficacy of lenvatinib in combination with pembrolizumab in patients with advanced urothelial cancer.MethodsIn this multicenter, open-label study (NCT02501096), patients with confirmed metastatic urothelial cancer and an ECOG PS of 0 or 1 received lenvatinib 20 mg orally once daily and 200 mg pembrolizumab intravenously every 3 weeks. Patients were not preselected based on PD-L1 status. The phase 2 primary end point was ORR at week 24 (ORRwk24), as assessed by study investigators using immune-related RECIST (irRECIST). Secondary end points included ORR, duration of response (DOR), and progression-free survival (PFS).ResultsAt the time of data cutoff , 20 patients were enrolled. 9 (45%) Patients were PD-L1(+); 5 (25%) were PD-L1(-); 6 (30%) were not tested. 4 Patients (20%) were treatment-naïve, whereas 11 (55%) and 5 (25%) patients had had 1 and 2 lines of prior anticancer therapies, respectively. No patient had received prior IO therapy. The primary end point of ORRwk24 was 25% (95% CI: 8.7–49.1). Additional efficacy outcomes are summarized in the table (Table 1). 18 (90%)Patients experienced treatment-related adverse events (TRAEs).Grade 3 and 4 TRAEs occurred in 5 (25%) and 5 (25%) patients, respectively. There was 1 fatal TRAE . The most common any-grade TRAEs were proteinuria (45%), diarrhea (40%), fatigue (30%), hypertension (30%), and hypothyroidism (30%).ConclusionsThe tyrosine kinase inhibitor (lenvatinib) and immunotherapy (pembrolizumab) regimen demonstrated activity in this study, which included patients receiving later-line treatment. The combination of lenvatinib and pembrolizumab deserves further investigation in patients with metastatic urothelial cancer.Trial RegistrationNCT02501096References1. Bellmunt J, et al. Pembrolizumab as second-line therapy for advanced urothelial carcinoma. N Engl J Med. 2017;376(11):1015-26.2. Vuky J et al. Updated efficacy and safety of KEYNOTE-052: A single arm phase 2 study investigating first-line pembrolizumab (pembro) in cisplatin-ineligible advanced urothelial cancer (UC). J Clin Oncol. 2018;36(15 Suppl):4524.3. Keytruda® (pembrolizumab) [package insert]. Whitehouse Station, NJ: Merck & Co., Inc.; 2017.Ethics ApprovalThis study was approved by all relevant institutional review boards.Table 1 (abstract P393). See text for description.P569Novel Pharmacobiotic approach to enhance the tamoxifen efficacy using bacterial extracellular vesicles as the immunotherapy in breast cancer1, Yeun-yeoul Yang1, Won-Hee Lee2, Jinho Yang2, Jong-kyu Kim1, HyunGoo Kim1, Se Hyun Paek1, Jun Woo Lee1, Joohyun Woo1, Jong Bin Kim1, Hyungju Kwon1, Woosung Lim1, Nam Sun Paik1, Yoon-Keun Kim2, Byung-In Moon1Jeongshin An, MD,PhD1Ewha Womans University, Seoul, Korea, Republic of; 2MD healthcare company, Seoul, Korea, Republic ofCorrespondence: Jeongshin An (rulru81@hanmail.net) Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P569Byung-In Moon is a contributing author and has therefore been added to the author list in this correction article.BackgroundThe anti-cancer effect of bacteria has a long history. According to Bierman et al., spontaneous remission of cancer has been observed in patients with severe bacteremia [1]. The reason was not revealed at that time, but we studied that in breast cancer. There are four main ways in which microbiota affects cancer: probiotics, prebiotics, drugs that target microbial enzymes and microbial products that have anticancer properties [2]. Among them, bacterial extracellular vesicles(EVs) are one of microbial products. In this study, we investigated the effects of bacterial EVs on the growth of breast cancer cells and tamoxifen efficacy.MethodsHere, we analized microbiota of urine samples by NGS to select the target EVs that were expected to affect the growth of breast cancer cells. A total of 347 female urine samples – from 127 breast cancer patients (cancer group) and 220 normal individuals (control group) – were collected and analyzed by NGS using a universal bacterial primer of 16S rDNA. Human breast cancer cells were cultured, and the cells were treated with EVs of S. aureus and K.pneumoniae for 72 h. Real-time polymerase chain reaction (PCR) and Western blotting for signalling molecule analysis were performed after treatment of EVs in each breast cancer cell.ResultsThere was a significant difference in the distribution of bacterial EVs between the urine samples from breast cancer patients and from normal controls. Especially, S.aureus EVs were predominant in the normal group, and K.pneumoniae was abundant in the breast cancer group. Therefore, we selected these two bacterial EVs that may have an effect on breast cancer cell growth. We found that S.aureus and K.pneumoniae EVs down-regulated cell growth in MDA-MB-231 cells. We also found that S.aureus or K.pneumoniae EVs had a synergic effect on growth inhibition of while co-treated with tamoxifen. S.aureus EVs down-regulated mRNA expression of cyclin E2 and up- regulated that of TNF-alpha which was related ERK pathway while co-treated with tamoxifen.ConclusionsThe anti-cancer effect of S.aureus and K.pneumoniae was initiated by its bacterial EVs and consequently inhibited the growth of breast cancer cells in triple negative breast cancer cells and improved the efficacy of tamoxifen in ER-positive cells. In the near future, we plan to conduct animal studies which are expected to further clarify the effect of bacterial EV on breast cancer. Ethics ApprovalThe study was approved by Ewha Womans University Medical Center‘s Ethics Board.ConsentWritten informed consent was obtained from the patient for publication of this abstract and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.References1. Bierman, H. R. et al. Remissions in leukemia of childhood following acute infectious disease. Staphylococcus and streptococcus, varicella, and feline panleukopenias. Cancer 6, 591-605 (1953).2. Zitvogel, L., Daillère, R., Roberti, M. P., Routy, B. & Kroemer, G. Anticancer effects of the microbiome and its products. Nature Reviews Microbiology 15, 465 (2017).Fig. 1 (abstract P569). See text for descriptionFig. 2 (abstract P569). See text for descriptionFig. 3 (abstract P569). See text for descriptionFig. 4 (abstract P569). See text for descriptionFig. 5 (abstract P569). See text for descriptionFig. 6 (abstract P569). See text for descriptionFig. 7 (abstract P569). See text for descriptionP651First-in-human study of FAZ053, an anti-PD-L1 mAb, alone and in combination with spartalizumab, an anti- PD-1 mAb, in patients with advanced malignancies2, David Tan3, Juan Martin-Liberal4, Shunji Takahashi5, Ravit Geva, MD6, Ayca Gucalp7, Xueying Chen8, Kulandayan Subramanian9, Jennifer Mataraza9, Jennifer Wheler9, Philippe Bedard, MD10Filip Janku, MD, PhD2MD Anderson Cancer Center, Houston, TX, USA; 3National University Cancer Institute, Singapore, Singapore 4Vall d’Hebron Institute of Oncology, Barcelona, Spain; 5The Cancer Institute Hospital of JFCR, Tokyo, Japan; 6Tel Aviv Sourasky Medical Center, Tel-Aviv, Israel; 7Memorial Sloan Kettering Cancer Center, New York, NY, USA 8Novartis Pharmaceuticals Corporation, East Hanover, NJ, USA 9Novartis Institutes for BioMedical Resea, Cambridge, MA, USA; 10Princess Margaret Cancer Centre, Toronto, ON, CanadaCorrespondence: Filip Janku (fjanku@mdanderson.org)Journal for ImmunoTherapy of Cancer 2018, 6(Suppl 1):P651Janis Callister was not a contributing author and has therefore been removed from the author list in this correction article. The redundant affiliation Articulate Science as shown on the original article is no longer listed on this correction article.BackgroundFAZ053 and spartalizumab are humanized immunoglobulin G4 monoclonal antibodies (mAbs) that bind anti-programmed death ligand-1 (PD-L1) and programmed death-1 (PD-1), respectively. We report the dose-escalation results from an ongoing Phase I study of FAZ053 ± spartalizumab in patients with advanced malignancies, enriched for patients with chordoma, a rare subtype of sarcoma.MethodsPatients received escalating doses of single-agent (SA) FAZ053 intravenously once every 3 weeks (Q3W) or 6 weeks (Q6W), or FAZ053 + spartalizumab Q3W. The primary objective was to assess the safety and tolerability of FAZ053 ± spartalizumab, and determine recommended doses for expansion (RDEs). Dose escalation was guided by an adaptive Bayesian logistic regression model following the escalation with overdose control principle.ResultsAs of the data cutoff of March 30, 2018, 61 patients received SA FAZ053 at doses 80–1600 mg Q3W or 800–1600 mg Q6W. Most patients  received prior treatment; 1 (2%) received prior anti-PD-1. FAZ053 exposure was generally dose proportional, with terminal half-life of ~16–18 days. A dose-limiting toxicity occurred in 1 patient . RDE was determined to be 1200 mg Q3W or 1600 mg Q4W. Adverse events (AEs) of all grades assessed as possibly related to treatment were reported for 33 patients (54%); most commonly (≥10%) fatigue  and pruritus ; 4 patients (7%) had Grade 3/4 treatment-related AEs, including elevated amylase (3%), renal failure, elevated lipase, elevated AST, and elevated blood CPK (each 2%). For these patients treated with SA FAZ053, partial responses (PRs) were demonstrated in 4 patients (7%) with chordoma, alveolar soft part sarcoma (ASPS), poorly differentiated carcinoma of scalp, and penile squamous cell carcinoma . Among 5 patients with chordoma treated with SA FAZ053, 1 patient has a PR, treatment ongoing >12 months, and 4 patients have stable disease ongoing (+4% to –29%). Data for 57 patients treated with combination FAZ053 (20–1200 mg) + spartalizumab 300 mg Q3W are preliminary. Updated results and biomarker data for patients receiving SA and combination treatment, including additional patients with chordoma, will be presented.ConclusionsSA FAZ053 was well tolerated and the RDE was determined to be 1200 mg Q3W. Clinical activity was observed in a range of indications including chordoma, a rare tumor without standard therapy options.Trial Registrationwww.clinicaltrials.gov; NCT02936102


Scientific Reports 10.1038/s41598-018-25836-4, published online 10 May 2018Correction to: In this Article, the legend of Figure 3 is incorrect:“A flowchart of the rapid (RAP) system for bone staining (RAP-B). The new procedure for bone staining which is rapid, nondestructive, and allowed to obtain high-definition, fine bone-stained specimens, was based on our RAP system. Cartilage staining (RAP-C) can be applied as single staining or optionally added before the bone staining for double staining of bone and cartilage . FIXATIVE: 5% formalin, 5% Triton X-100, 1% potassium hydroxide (KOH); B-STAINING SOLUTION: 0.05% alizarin red S, 20% ethylene glycol, 1% KOH; C-STAINING SOLUTION: 50–70% ethanol, 20% acetate, 0.015–0.02% alcian blue; CLEARING SOLUTION: 20% Tween 20, 1% KOH; ENHANCEMENT SOLUTION: 20% ethylene glycol, 5% Triton X-100, 1% KOH.”should read:“A flowchart of the rapid (RAP) system for bone staining (RAP-B). The new procedure for bone staining which is rapid, nondestructive, and allowed to obtain high-definition, fine bone-stained specimens, was based on our RAP system. Cartilage staining (RAP-C) can be applied as single staining or optionally added before the bone staining for double staining of bone and cartilage . FIXATIVE: 5% formalin, 5% Triton X-100, 1% potassium hydroxide (KOH); B-STAINING SOLUTION: 0.05% alizarin red S, 20% ethylene glycol, 1% KOH; C-STAINING SOLUTION: 50–70% ethanol, 20% acetic acid, 0.015–0.02% alcian blue; CLEARING SOLUTION: 20% Tween 20, 1% KOH; ENHANCEMENT SOLUTION: 20% ethylene glycol, 5% Triton X-100, 1% KOH.”


Plasmodium vivax infections. Knowledge of the metabolism of PQ is critical for understanding the therapeutic efficacy and hemolytic toxicity of this drug. Recent in vitro studies with primary human hepatocytes have been useful for developing the ultra high-performance liquid chromatography coupled with high-resolution mass spectrometric (UHPLC-QToF-MS) methods for simultaneous determination of PQ and its metabolites generated through phase I and phase II pathways for drug metabolism.Primaquine (PQ), an 8-aminoquinoline, is the only drug approved by the United States Food and Drug Administration for radical cure and prevention of relapse in These methods were further optimized and applied for phenotyping PQ metabolites from plasma and urine from healthy human volunteers treated with single 45 mg dose of PQ. Identity of the metabolites was predicted by MetaboLynx using LC–MS/MS fragmentation patterns. Selected metabolites were confirmed with appropriate standards.Besides PQ and carboxy PQ (cPQ), the major plasma metabolite, thirty-four additional metabolites were identified in human plasma and urine. Based on these metabolites, PQ is viewed as metabolized in humans via three pathways. Pathway 1 involves direct glucuronide/glucose/carbamate/acetate conjugation of PQ. Pathway 2 involves hydroxylation (likely cytochrome P450-mediated) at different positions on the quinoline ring, with mono-, di-, or even tri-hydroxylations possible, and subsequent glucuronide conjugation of the hydroxylated metabolites. Pathway 3 involves the monoamine oxidase catalyzed oxidative deamination of PQ resulting in formation of PQ-aldehyde, PQ alcohol and cPQ, which are further metabolized through additional phase I hydroxylations and/or phase II glucuronide conjugations.This approach and these findings augment our understanding and provide comprehensive view of pathways for PQ metabolism in humans. These will advance the clinical studies of PQ metabolism in different populations for different therapeutic regimens and an understanding of the role these play in PQ efficacy and safety outcomes, and their possible relation to metabolizing enzyme polymorphisms.The online version of this article (10.1186/s12936-018-2433-z) contains supplementary material, which is available to authorized users. Plasmodium vivax malaria + (intense peak), 175.0842 [M + H-C5H8O2]+, 101.0593 [M + H-C10H10N2O]+, 83.0493 [M + H-C10H12N2O2]+ , 260.177 , 243.1523 , 175.0895 , 86.0959  , 160.0619 [M + H-C8H16NO2]+, 88.0384 [M + H-C14H18N2O]+ , 147.0553 [M + H-C6H11NO]+, 86.0970 [M + H-C9H6N2O2]+ , 2-Hydroxy PQ (18) and 4-hydroxy PQ (19) with the same retention time (Table 17) could not be established due to non-availability of corresponding synthetic standard.ESI-HRMS of these four metabolites gave identical molecular ion peak at m/z 453.1881 [M + H]+ which corresponds to the molecular formula C21H29N2O9. , 161.0751 [M + H-C5H11N]+, 86.0966 [M + H-C9H8N2O]+, 69.0721 [M + H-C9H11N3O]+ , 86.0966 [M + H-C12H12N2O2]+ , 175.0884 [M + H-C11H16O8]+, 101.0593 [M + H-C16H18N2O7]+  (2) and cPQ (m/z 275.1385) (5), only seven additional metabolites of PQ were detected in plasma. These include four phase I metabolites namely, hydroxy-primaquine quinone-imine (m/z 274.1549) (1), carboxyprimaquine lactam (m/z 257.1273) (3), Dihydroxy-carboxyprimaquine methylester (m/z 321.1454) (8) and carboxyprimaquine methyl ester (m/z 289.1560) (6) and three phase II metabolites, namely, glycosylated PQ (m/z 422.2315) (7), PQ N-carbamoyl glucuronide (m/z 480.1992) (4) and PQ methyl carbamate (m/z 318.1789) (9). No hydroxylated metabolites of PQ were detected in plasma.Very limited numbers of PQ metabolites were detected in plasma from the individuals treated with single dose of PQ. Besides PQ (m/z 260.1769) (2) and the major plasma metabolite cPQ (m/z 275.1385) (5) were detected in urine also. Besides these, the carboxy PQ lactam (m/z 257.1273) (3), predictably formed by spontaneous cyclization of carboxy side chain of cPQ and PQ carbamoyl glucuronide (m/z 480.1992) (4), a unique phase II metabolite formed through direct N-carbamoylation of PQ followed by glucuronidation, which were present in plasma, were also detected in urine. Glycosylated PQ (m/z 422.2315) (7), a prominent metabolite detected in plasma, was not detected in urine. Three acetylated metabolites were present in urine namely, PQ acetate (m/z 302.1875) (29), acetylated hydroxyl-PQ glucuronide conjugate (m/z 494.2127) (20) and N-Acetyl-PQ quinone-imine (m/z 316.1676) (35). Two metabolites with different retention times but identical accurate mass and MS/MS fragmentation profile corresponding to PQ glucuronide conjugate (m/z 436.2100)  were also detected in urine.The urine from the individuals treated with PQ contained large numbers of PQ metabolites. PQ (m/z 276.1694) correspond to 2-OH (18), 3-OH (21), and 4-OHPQ (14), and the fourth (17) could be 7-OH, 8-N–OH, 1-N-oxide, or 8-N-oxide of PQ. Desmethy-PQ (m/z 246.1606) (19), and desmethyl-PQ glucuronide (m/z 422.1908) (12) were also detected in urine. Two metabolites predicted as PQ-quinone-imine (m/z 274.1549)  and PQ-5,6-ortho-quinone (m/z 260.1403) (10), which is spontaneously generated from 5-OH-PQ, were also detected in urine. None of these phase I metabolites were detected in plasma. Further metabolism of hydroxyl-PQ was evident from presence of two dihydroxy-PQ (m/z 292.1647) metabolites  and a dihydroxyl PQ quinone-imine (m/z 290.15) (24). Phase II conjugation metabolites, generated from further metabolism of hydroxyl PQ metabolites, detected in urine were two monohydroxy-PQ glucuronides (m/z 452.2011)  trihydroxy-PQ glucuronide (m/z 484.1932) (23) and desmethyl-PQ glucuronide (m/z 422.1908) (12).Surprisingly, four mono hydroxylated metabolites of PQ, likely to be formed through CYP mediated pathways, were detected in urine. Three of these metabolites (m/z 451.1722) (32), hydroxyl cPQ glucuronide (m/z 467.1649) (27), trihydroxy-cPQ glucuronide (m/z 499.1545) (26) and dihydroxy cPQ methylester (m/z 321.1454) (8). Hydroxy-PQ alcohol (m/z 277.1559) (33), PQ alcohol glucuronide (m/z 437.1911) (31), hydroxyl-PQ alcohol glucuronide (m/z 453.1881) (15) detected in urine may be generated through further metabolism of PQ alcohol, the metabolite from oxidative deamination of PQ side chain alkyl amine.Further metabolism of cPQ, a major plasma metabolite, through phase I and phase II pathways of drug metabolism was evident from presence of several cPQ metabolites namely, cPQ glucuronide  [N-carbamoylation and glucuronidation [2 tension yield N-carbamoyl glucuronide conjugates [N-carbamoyl glucuronide conjugate of PQ as a predominant metabolite, when PQ was incubated in vitro with primary human hepatocytes [S-PQ generated more than two-fold higher levels of the conjugate than (−)-R-PQ [R-PQ to the oxidative deamination pathway [S-PQ than from (−)-R-PQ. In the human hepatocyte study, glycosylated PQ was detected at time zero and thought to be a simple adduct of the primary amine group of PQ and the aldehyde form of glucose via non-enzymatic means. This metabolite could also be formed in plasma by the same mechanism. Predominant metabolism of (+)-S-PQ through these pathways may explain the somewhat lower plasma concentrations of the (+)-S-PQ, despite its negligible metabolism through oxidative deamination to cPQ [Based on the comprehensive LC-ESI-HRMS and MS/MS fragmentation profiles of PQ metabolites identified in plasma and urine of healthy human volunteers treated with single dose of 45 mg, PQ may be regarded as metabolized broadly through three pathways. Pathway 1 involves direct glucuronide/glucose/carbamate/acetate conjugation of PQ. The presence of s (UGTs) . Althougnidation , 36. Incatocytes . Formati(−)-R-PQ . This ma pathway . Glycosy pathway  and was n to cPQ , 20.orthoquinone was only detected in urine but not in plasma. Recent PK and tissue distribution studies in mice showed rapid appearance of PQ-5,6-orthoquinone in high concentrations in the liver and in low concentration in plasma and its fast disappearance from the liver and circulation [orthoquinone PQ. Earlier in vitro studies have indicated the role of multiple CYP isoforms in hemolytic toxicity of PQ with predominant involvement of CYP2D6 and CYP3A4 [orthoquinone metabolite, has been linked to therapeutic efficacy of PQ [orthoquinone through CYP2D6 mediated pathway [The pathway 2  mediated pathways, generates monohydroxylated PQ metabolites. These metabolites have been implicated in therapeutic efficacy and hemolytic toxicity of PQ. Further, extension of these studies to quantitative profiling of key metabolites in malaria infected individuals treated with PQ and their correlation with therapeutic outcomes vis-à-vis CYP2D6 genotypes may be useful in determining the rational approach for application of PQ in malaria radical cure, the primary application of this drug, as well as malaria control programs.This is the most exhaustive study on identification and characterization of human metabolites of primaquine, the only approved drug for malaria radical cure. Based on overall metabolites’ profile, three pathways were predicted for metabolism by humans, (a) direct glucuronide/glucose/carbamate/acetate conjugation of PQ (b) hydroxylation of PQ at different positions on the quinoline ring and (c) oxidative deamination of PQ at the terminal amine of the aminoalkyl side chain. Metabolism of PQ through direct conjugation and oxidative deamination pathways may determine pharmacokinetics and pharmacodynamics of this important antimalarial drug. Metabolism of PQ through hydroxylations on the quinolone ring, predictably catalyzed by cytochrome PAdditional file 1: Fig. 1S. Key fragments of PQ and other metabolites using LC/ESI-QToF


This article has been corrected: The authors made a mistake in size of scale bar in captions of Figures 2-5. The authors declare that this correction does not change the results or conclusions of this paper. The authors sincerely apologize for this error. The correct Figure captions are provided below. The correct scale bar size is marked in bold font.Ink4a+/CD34+) in human skeletal muscle after resistance exercise.Figure 2. Senescent endothelial progenitor cells (p16 (A) Representative immunohistochemical co-staining of muscle cross-sections (senescent cells indicated by arrows). Scale bar 55 μm. (B) Senescent endothelial progenitor cells decreased in human skeletal muscle after a single bout of resistance exercise, and to a greater extends under low protein supplemented condition. * Significant difference against Baseline, P < 0.05; † Significant difference against Low Protein, P < 0.05. Low protein: 14% protein; High protein: 44% protein in weight.+) in human skeletal muscle after resistance exercise.Figure 3. Phagocytic macrophage (CD68 (A) Representative immuno-histochemical staining of a muscle cross-section (CD68+ macrophage indicated by an arrow). Scale bar 100 μm. (B) Low protein supplementation before and after resistance exercise enhanced CD68+ macrophage infiltration in skeletal muscle above High protein trial. * Significant difference against Baseline, P < 0.05; † Significant difference against Low Protein, P < 0.05. Low protein: 14% protein; High protein: 44% protein in weight.+) in human skeletal muscle after resistance exercise.Figure 4. Regenerative macrophage (CD163 (A) Representative immunohistochemical staining of a muscle cross-section (CD163+ macrophage indicated by an arrow). Scale bar 100 μm. (B) High protein supplementation before and after resistance exercise increased CD163+ macrophage presence in human skeletal muscle 48 h after exercise. * Significant difference against Baseline, P < 0.05; † Significant difference against Low Protein, P < 0.05. Low protein: 14% protein; High protein: 44% protein in weight.Figure 5. Centrally nucleated fibers in human skeletal muscle after resistance exercise. (A) Representative hematoxylin and eosin staining of a muscle cross-section . Scale bar 100 μm. (B) (††) No difference between Low and High protein trials was found. *Significant difference against Baseline, P < 0.05. Low protein: 14% protein; High protein: 44% protein in weight.1356-1365 . https://doi.org/10.18632/aging.101472Original article: Aging. 2018; 10:1356–1365.


Streptococcus pneumoniae Serotype 15A-ST63 Clone in Japan, 2012–2014 . The article has been corrected online (https://wwwnc.cdc.gov/eid/article/24/2/17-1276_article).Some data were inaccurate in the text and figures in Spread of Meropenem-Resistant


The Supporting Information files were published with tracked changes. Please view the correct files below.S1 Table(DOCX)Click here for additional data file.S2 Table(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.S7 Table(DOCX)Click here for additional data file.S1 Fig(DOCX)Click here for additional data file.S2 Fig(DOCX)Click here for additional data file.S3 Figvs. progressors using eGFR , Gd-IgA1 biomarkers, and Oxford classification . Area under the curve, AUC = 0.936. b- Receiver operating characteristic (ROC) curve for non-progressors vs. progressors using eGFR  and Oxford classification . Area under the curve, AUC = 0.836.a- ROC curve for non-progressors (DOCX)Click here for additional data file.S4 Fig2 at the time of renal-biopsy; group 2 (n = 42), eGFR <60 mL/min/1.73 m2 at the time of renal-biopsy. Prediction equation from logistic regression (predicts probability to choose group 1): Pred(group 1) = 1 / (1 + exp(-(1517.5–1.2E-02*AB-IgA-24.9*eGFR))).Area under the curve = 1.00. Accuracy of the discrimination is 100%. Group 1 (n = 35), eGFR ≥60 mL/min/1.73 m(DOCX)Click here for additional data file.S5 FigS-creat, serum creatinine (μmol/L); eGFR ; serum IgG autoantibody specific for Gd-IgA1 (U/mL). Group 1 (n = 35), eGFR ≥60 mL/min/1.73 m2 at the time of renal biopsy; group 2 (n = 42), eGFR <60 mL/min/1.73 m2 at the time of renal biopsy.(DOCX)Click here for additional data file.


Scientific Reports 10.1038/s41598-018-34127-x, published online 23 October 2018Correction to: This Article contains errors in Figure 1, where 4 microbial diversity patterns were omitted. The correct Fig. As a result, the Figure legend,A. mali gut microbiotas. Alpha-diversity in bacterial  and fungal  gut communities of A. mali gut. Beta-diversity in bacterial (E) and fungal (F) gut communities estimated via principal coordinate analysis (PCoA) based on Bray-Curtis distance.”“Microbial diversity patterns for should read:A. mali gut microbiotas. Alpha-diversity in bacterial  and fungal  gut communities of A. mali gut. Beta-diversity in bacterial (E) and fungal (F) gut communities estimated via principal coordinate analysis (PCoA) based on Bray-Curtis distance.”“Microbial diversity patterns for


Scientific Reports 10.1038/s41598-019-54905-5, published online 05 December 2019Correction to: This Article contains a typographical error in the Acknowledgements section.“Ministry of Education and Science of Russian Federation (Grant № 12.1223.2017/AP to S.S. and E.R.).”should read:“Ministry of Education and Science of Russian Federation (Grant No. 18-15-00172 to S.S.and E.R.)”


Acinetobacter radioresistens strain DD78 (= CCUG 69565) is a soil hydrocarbon-degrading and biosurfactant-producing bacterium isolated from chronically crude oil-polluted soil of the Aconcagua River mouth in Chile. The 3.25-Mb A. radioresistens DD78 genome (41.8% GC content) was completely sequenced, with 4 replicons, 2,970 coding sequences, and 77 tRNAs. Acinetobacter radioresistens strain DD78 (= CCUG 69565) is a soil hydrocarbon-degrading and biosurfactant-producing bacterium isolated from chronically crude oil-polluted soil of the Aconcagua River mouth in Chile. The 3.25-Mb A. radioresistens DD78 genome (41.8% GC content) was completely sequenced, with 4 replicons, 2,970 coding sequences, and 77 tRNAs. Acinetobacter species are aerobic Gram-negative coccobacilli present in soil, freshwater, sediment, polluted environments, and clinical samples , improving hydrocarbon bioavailability (A. radioresistens DD78 (CCUG_69565), a soil hydrocarbon-degrading and biosurfactant-producing bacterium from crude oil-polluted soil of the Aconcagua River mouth in Central Chile, which was isolated using enrichment in diesel (0.1% [vol/vol]) in Bushnell Hass mineral medium . The PacBio reads were trimmed and assembled into 5 contigs using SMRTLink v5 and the Hierarchical Genome Assembly Process v4.0 (HGAP4.0) with default parameters , proteins for osmoprotectant glycine betaine synthesis , and the osmoprotectant transporters osmo-dependent choline transporter, sodium/proline symporter, and aspartate/alanine antiporter  were identified (>30% identity) with BLASTP using the Uniprot-KB/Swiss-Prot database. Arsenic and heavy metal  resistance genes are in pAr1. Catabolic genes encoding an alkane monooxygenase , a rubredoxin NAD(H) reductase/rubredoxin system (E3H47_10395 and E3H47_10400), and the catechol  and benzoate catabolic pathways  are chromosomal. Comparative complete 16S rRNA gene sequence analysis showed a close relationship (99.7% identity) with A. radioresistens FO-1T  and <97.5% identity with other Acinetobacter species (A. radioresistens CIP 103788T, A. lwoffii ATCC 9957T, and A. tandoii CIP 107469T showed ANIb values of 98.9%, 74.1%, and 73.6%, respectively, indicating that strain DD78 most likely belongs to the species A. radioresistens. The genome sequence of A. radioresistens DD78 provides essential data on alkane degradation, salt resistance, and biosurfactant production. samples , 2. A. b alkanes , 4. A. rlability , 6. Here78 CCUG_6565, a some sizes . The 5 cme sizes . The DD7me sizes  identifime sizes  identifi isolate . Genes e species  with A. Acinetobacter radioresistens DD78 genome sequences have been deposited in DDBJ/ENA/GenBank under the accession number GCA_005519305 and the SRA accession number SRX5548971. The version discussed here is the first version. The BioProject number for the publicly available raw data is PRJNA528312.The


Blood cultures were negative. Laparoscopic nodal biopsy showed sheets of medium-sized lymphocytes diffusely expressing CD30, TIA-1, granzyme B, and ALK, but not T-cell markers including CD2, CD3, CD4, CD5, CD7, CD8, and βF1, indicating ALK+ ALCL of null cell phenotype. Bone marrow biopsy showed two small aggregates of tumor cells in a background of normal tri-lineage hematopoiesis. ALK immunostaining revealed singly scattered positive cells (Anaplastic large cell lymphoma (ALCL) frequently involves both nodal and extranodal sites and is rarely leukemic. A 21-year-old male presented with abdominal pain. His complete blood count, which had been normal four months ago, showed increasing white cell counts from 14.9x10ve cells  in additve cells . The dis


Correction: J Intensive Carehttps://doi.org/10.1186/s40560-018-0352-2In the original publication of this article , an AuthSome data concerning specifically ECMO-treatment in patients with septic shock have been published previously in a German journal with respect to a completely different research question: Friedrichson B, Fichte J, Banjas N, Schütz M, Hopf H-B. Extrakorporale Membranoxygenierung bei erwachsenen Patienten mit septischem Schock. Anästhesie und Intensivmed. 2018;59:698–704.


Haemophilus influenzae and Streptococcus pneumoniae has been implicated in the pathogenesis of otitis media, mainly in chronic and recurrent cases. We studied the “in vitro” biofilm production by these 2 species isolated alone or together from the nasopharynx of children with acute otitis media.Biofilm production by H. influenzae and 191 S. pneumoniae strains.The studied strains were from 3 pneumococcal conjugate vaccine (PCV) periods: pre-PCV7, post-PCV7/pre-PCV13 and post-PCV13. A modified microtiter plate assay with crystal violet stain was used to study the biofilm production of 182 H. influenzae and 128/191 (66.8%) S. pneumoniae strains produced biofilm. The proportion of biofilm-producing H. influenzae strains was greater with than without the isolation of S. pneumoniae in the same sample . Conversely, the proportion of biofilm-producing S. pneumoniae strains was not affected by the presence or not of H. influenzae (66.3% vs 67.4%). S. pneumoniae serotypes 6B, 15B/C, 19A, 35F and 35B were the better biofilm producers (80%). Serotypes 11A, 14, 15A, 19F and 19A were more associated with H. influenzae biofilm-producing strains. Overall, 89/94 (94.6%) of cases with combined isolation showed biofilm production by S. pneumoniae or H. influenzae.Overall, 117/181 (64.6%) H. influenzae and S. pneumoniae strains isolated from the nasopharynx of children with acute otitis media, which reinforces the results of studies suggesting the importance of biofilm in the pathogenesis of acute otitis media.This study emphasizes the high proportion of biofilm production by The online version of this article (10.1186/s12879-018-3657-9) contains supplementary material, which is available to authorized users. Acute otitis media (AOM) is the leading cause of bacterial infections in childhood, about 700 million cases each year, and the leading cause of antibiotic prescription . More thStreptococcus pneumoniae and non-typable Haemophilus influenzae (NT-Hi)  as well as Moraxella catarrhalis : strong production > 1.10, moderate production 0.35–1.10, negative < 0.35.The cut-offs used to determine the intensity of biofilm production in isolates of S. pneumoniae  were classified as strong biofilm producers, 55 (30.2%) moderate producers and 64 (35.1%) non-producers. One strain could not be evaluated. Overall, 117/181 (64.6%) NT-Hi strains produced biofilm  were classified as strong biofilm producers, 64 (33.5%) moderate producers and 63 (33%) non-producers. Overall, 128/191 (66.8%) S. pneumoniae strains produced biofilm , whether isolated alone or with S. pneumoniae strains  or with S. pneumoniae strains . Overall, the proportion of NT-Hi biofilm-producing strains was greater when isolated with S. pneumoniae   or with NT-Hi strains (71.9 and 62.5% in pre-PCV7 and post-PCV13 periods). Overall, the proportion of S. pneumoniae biofilm-producing strains did not differ whether isolated alone or with NT-Hi strains   that produced biofilm was similar to that for other serotypes: 57.4% (31/54) versus 70.6% (96/137), p = 0.09 (data not shown). Moreover, the proportion did not differ between the 6 additional S. pneumoniae serotypes included in PCV13 and other serotypes . Strains of serotypes 6B, 15B/C, 19A, 35F, and 35B produced biofilm in more than 80% of the cases. In contrast, strains of serotypes 23B, 23F,19F were the lowest producers (40% of strains)  of the combination. H. influenzae biofilm-producing strains were isolated more often with serotypes 11A, 14, 15A, 19F and 19A. This association did not affect biofilm production by S. pneumoniae.The p = 0.44.Susceptibility or resistance to penicillin did not differ with and without biofilm production: 64.2% (61/96) versus 69.5% (66/95), -Hi strains produced biofilm when isolated with S. pneumoniae strains. Serotypes of S. pneumoniae were not associated with biofilm production by NT-Hi strains.Overall, 75.5% (72/96) of NTS. pneumoniae or H. influenzae or both produced biofilm in 94.6% (89/94) of cases  or fever + otalgia . These relations were not observed for S. pneumoniae .Table -Hi and S. pneumoniae. Among NT-Hi strains, the proportion of β-lactamase–producing strains decreased from 51.5% (33/64) in the pre-PCV7 period to 17.8% (10/56) and 17.7% (11/62) in the post-PCV7/pre-PCV13 and post-PCV13 periods, respectively (p < 0.001). As well, the proportion of S. pneumoniae strains with decreased susceptibility to penicillin decreased from 65.7% in the pre-PCV7 period to 45.4% (29/64) and 39.7% (25/63) in the post-PCV7/pre-PCV13 and post-PCV13 periods (p = 0.003). Among S. pneumoniae strains, the proportion of those with erythromycin susceptibility increased from 46.8% (30/64) to 81% (51/63) between the post-PCV7/pre-PCV13 and post-PCV13 periods, respectively (p < 0.001).The proportion of resistant strains significantly decreased from the pre-PCV7 to post-PCV13 periods for both NT. pneumoniae serotypes changed according to the study period  with no variation in proportion for S. pneumoniae over PCV implementation periods. Of note, the proportion of NT-Hi biofilm-producing strains was greater when H. influenzae strains were isolated with S. pneumoniae, which agreed with the results of Hong et al. : strong production > 1.10, moderate production 0.35–1.10, negative < 0.35. Strains were classified according to agreement of at least 2 of the 3 methods used to calculate biofilm production. (XLS 195 kb)Supplementary Data S1: Different cut-offs used to determine the biofilm production by Additional file 2:H. influenzae: the different cut-offs used were as followed where AB represented the stained wells containing attached bacteria, CW the stained control wells containing bacteria-free medium only and G the bacterial growth control i) AB – CW: strong production > 0.30, moderate production 0.10–0.30, negative < 0.10 ii) AB / CW: strong production > 6, moderate production 2–6, negative < 2 iii)[(AB – CW) / G]: strong production > 1.10, moderate production 0.35–1.10, negative < 0.35. Strains were classified according to agreement of at least 2 of the 3 methods used to calculate biofilm production. (XLSX 76 kb)Supplementary Data S2: Different cut-offs used to determine the biofilm production by Additional file 3:. pneumoniae serotypes by study period . (DOCX 15 kb)Supplementary Data S3: Distribution of S


Scientific Reports 10.1038/s41598-017-01512-x, published online 02 May 2017Correction to: This Article contains an error in Reference 29:PLoS One12(4), e0175267 (2017)’‘Jose Carlos Mancera Gracia, Silvie Van den Hoecke, Xavier Saelens, Kristien Van Reeth & Mirco Schmolke. Effect of serial pig passages on the adaptation of an avian H9N2 influenza virus to swine. should read:PLoS One12(4), e0175267 (2017)’‘Mancera Gracia, J. C., Van den Hoecke, S., Saelens, X. & Van Reeth, K. Effect of serial pig passages on the adaptation of an avian H9N2 influenza virus to swine.


Ralstonia solanacearum is a major phytopathogenic bacterium that attacks many crops and other plants around the world. In this study, a novel actinomycete, designated strain NEAU-SSA 1T, which exhibited antibacterial activity against Ralstonia solanacearum, was isolated from soil collected from Mount Song and characterized using a polyphasic approach. Morphological and chemotaxonomic characteristics of the strain coincided with those of the genus Streptomyces. The 16S rRNA gene sequence analysis showed that the isolate was most closely related to Streptomyces aureoverticillatus JCM 4347T (97.9%). Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain formed a cluster with Streptomyces vastus JCM4524T (97.4%), S. cinereus DSM43033T (97.2%), S. xiangluensis NEAU-LA29T (97.1%) and S. flaveus JCM3035T (97.1%). The cell wall contained LL-diaminopimelic acid and the whole-cell hydrolysates were ribose, mannose and galactose. The polar lipids were diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), hydroxy-phosphatidylethanolamine (OH-PE), phosphatidylinositol (PI), two phosphatidylinositol mannosides (PIMs) and an unidentified phospholipid (PL). The menaquinones were MK-9(H4), MK-9(H6), and MK-9(H8). The major fatty acids were iso-C17:0, C16:0 and C17:1 ω9c. The DNA G+C content was 69.9 mol %. However, multilocus sequence analysis (MLSA) based on five other house-keeping genes , DNA–DNA relatedness, and physiological and biochemical data showed that the strain could be distinguished from its closest relatives. Therefore, it is proposed that strain NEAU-SSA 1T should be classified as representatives of a novel species of the genus Streptomyces, for which the name Streptomyces sporangiiformans sp. nov. is proposed. The type strain is NEAU-SSA 1T (=CCTCC AA 2017028T = DSM 105692T). Ralstonia solanacearum is the causal agent of bacterial wilt, one of the most devastating plant pathogenic bacteria around the world [he world , which hhe world , includihe world . TherefoStreptomyces, within the family Streptomycetaceae, is the largest genus of the phylum Actinobacteria, first proposed by Waksman and Henrici (1943) [http://www.bacterio.net/streptomyces.html), which are widely distributed in soils throughout the world. Therefore, members of novel Streptomyces species are in demand as sources of novel, environmentally friendly, commercially significant, naturally bioactive compounds [T with inhibitory activity against phytopathogenic bacterium Ralstonia solanacearum was isolated and subjected to the polyphasic taxonomy analysis. Results demonstrated that the strain represents a novel species of the genus Streptomyces, for which the name Streptomyces sporangiiformans sp. nov. is proposed.The actinobacteria are known to produce biologically active secondary metabolites, including antibiotics, enzymes, enzyme inhibitors, antitumour agents and antibacterial compounds ,5,6. Thei 1943)  and currompounds ,9. Durin43  and cT was isolated from soil collected from Mount Song , Dengfeng, Henan Province, China. The soil sample was air-dried at room temperature for 14 days before isolation for actinomycetes. After drying, the soil sample was ground into powder and then suspended in sterile distilled water, followed by a standard serial dilution technique. The diluted soil suspension was spread on humic acid-vitamin agar (HV) [−1) and nalidixic acid (20 mg L−1). After 28 days of aerobic incubation at 28 °C, colonies were transferred and purified on the International Streptomyces Project (ISP) medium 3 [v/v) at −80 °C for long-term preservation.Strain NEAU-SSA 1gar (HV)  supplemew/v) were tested in GY (Glucose-Yeast extract) medium [2PO4/0.1 M NaOH; pH 9.0–10.0, 0.1 M NaHCO3/0.1 M Na2CO3; and pH 11.0–12.0, 0.2 M KH2PO4/0.1 M NaOH. Hydrolysis of Tweens  and production of urease were tested as described by Smibert and Krieg [2S were examined as described previously [Gram staining was carried out by using the standard Gram stain, and morphological characteristics were observed using light microscopy  and scanning electron microscopy  using cultures grown on ISP 3 agar at 28 °C for 6 weeks. Samples for scanning electron microscopy were prepared as described by Jin et al. . Cultura) medium  at 28 °Cnd Krieg . The utieviously ,20.18 Column  that had a mobile phase consisting of acetonitrile: 0.05 mol L−1 phosphate buffer pH 7.2  at a flow rate of 0.5 mL min−1. The peak detection used an Agilent G1321A fluorescence detector  with a 365 nm excitation and 455 nm longpass emission filters. The whole-cell sugars were analyzed according to the procedures developed by Lechevalier and Lechevalier [18 Column  at 270 nm. The mobile phase was acetonitrile-iso-propyl alcohol . To determine cellular fatty acid compositions, the strain NEAU-SSA 1T was cultivated in GY medium in shake flasks at 28 °C for 4 days. Fatty acid methyl esters were extracted from the biomass as described by Gao et al. [Biomass for chemotaxonomic studies was prepared by growing the organisms in GY medium in shake flasks at 28 °C for 5 days. Cells were harvested using centrifugation, washed with distilled water, and freeze-dried. The isomer of diaminopimelic acid (DPA) in the cell wall hydrolysates was derivatized and analyzed using an HPLC (High Performance Liquid Chromatography) method  with an hevalier . The polhevalier . Menaquihevalier . Extracto et al.  and analT was cultured in GY medium for 3 days to the early stationary phase and harvested using centrifugation. The chromosomal DNA was extracted according to the method of sodium dodecyl sulfate (SDS)-based DNA extraction [T (1412bp) was obtained and compared with type strains available at the EzBioCloud server (https://www.ezbiocloud.net/), retrieved using NCBI BLAST  and then submitted to the GenBank database. Phylogenetic trees were constructed based on the 16S rRNA gene sequences of strain NEAU-SSA 1T and related reference species. Sequences were multiply aligned in Molecular Evolutionary Genetics Analysis (MEGA) software version 7.0 using the Clustal W algorithm and trimmed manually where necessary. Phylogenetic trees were constructed with neighbor-joining [T to its closely related strains, phylogenetic relationships of the strain NEAU-SSA 1T were also confirmed using sequences of five individual housekeeping genes  for core-genome analysis. The sequences of NEAU-SSA 1T and its related strains were obtained from the genomes or GenBank/EMBL/DDBJ (European Molecular Biology Laboratory/DNA Data Bank of Japan). GenBank accession numbers of the sequences used are given in atpD-gyrB-recA-rpoB-trpB. Phylogenetic analysis was performed as described above. Genome mining for bioactive secondary metabolites was performed using “antibiotics and secondary metabolite analysis shell” (antiSMASH) version 4.0 [For DNA extraction, strain NEAU-SSA 1traction . PCR amptraction ,30. The -joining  and maxi-joining  algorith-joining . The sta-joining . A dista-joining . All pos-joining . To furtsion 4.0 .T was extracted using the method of SDS-based DNA extraction [® 2.0 Fluorometer (Thermo Scientific). Whole-genome sequencing was performed on the Illumina HiSeq PE150  platform. A-tailed, ligated to paired-end adaptors, and PCR amplified samples with a 350 bp insert were used for the library construction at the Beijing Novogene Bioinformatics Technology Co., Ltd. Illumina PCR adapter reads and low-quality reads from the paired-end were filtered using a quality control step using our own compling pipeline. All good-quality paired reads were assembled using the SOAP (Short Oligonucleotide Alignment Program) denovo [https://github.com/aquaskyline) into a number of scaffolds. Then, the filter reads were handled by the next step of the gap-closing.For draft genome sequencing and assembly, the genomic DNA of strain NEAU-SSA 1traction . The har) denovo ,39 , then sheared using a JY92-II ultrasonic cell disruptor . The DNA renaturation rates were determined in 2 × SSC at 70 °C. The experiments were performed with three replications and the DNA–DNA relatedness value was expressed as a mean of the three values. Several genomic metrics are now available to distinguish between orthologous genes of closely related prokaryotes, including the calculation of average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values [T and S. flaveus JCM3035T (JOCU00000000) using the ortho-ANIu algorithm from Ezbiotaxon and the genome-to-genome distance calculator (GGDC 2.0) at http://ggdc.dsmz.de.Because of a lacking number of genome sequences of y et al.  under coy et al. , using atraction . The con) values ,43. In tT against two pathogenic bacteria  was evaluated using the agar well diffusion method [T, the strain was cultured in tryptone-glucose-soluble starch-yeast extract medium , and the inhibitory activity was tested. Briefly, strain NEAU-SSA 1T was inoculated into MB medium and incubated at 28 °C for seven days in a rotary shaker. The supernatant (100 mL for this study) was obtained via centrifugation at 8000 rpm and 4 °C for 10 min and subsequently extracted by using an equal volume of ethyl acetate. Then, the extract was dried in a rotary evaporator at 40 °C and eluted with proper volume methanol (1 mL used in this study). The cell precipitate was extracted with an equal volume of methanol and also condensed as above. After that, the antibacterial activity was evaluated using the agar well diffusion method, and each well contained 200 µL of the methanol extract. To examine the effect of temperature on antibacterial activity, the ten-fold dilution methanol extract was placed in a water bath at 40, 60, 80, and 100 °C for 30 min, and then cooled to room temperature. The antibacterial activity was evaluated using the agar well diffusion method.The antibacterial activity of strain NEAU-SSA 1n method  with theT showed that the strain had the typical characteristics of the genus Streptomyces. Observation of 6-week cultures of strain NEAU-SSA 1T grown on ISP 3 medium revealed that it formed well-developed, branched substrate hyphae and aerial mycelia. Sporangia consisted of cylindrical, and rough-surfaced spores (0.6–0.8 μm × 0.9–1.6 μm) were produced on aerial mycelia, but spore chains were not observed  with an optimal level of 0–1% (w/v). Detailed physiological characteristics are presented in the species description  (29.5%), MK-9(H6) (41.2%), and MK-9(H8) (29.4%). The cellular fatty acid profile of strain NEAU-SSA 1T was composed of iso-C17:0 (30.9%), C16:0 (26.4%), C17:1ω9c (19.9%), C15:0 (7.8%), C17:0 (4.4%), C14:0 (3.3%), iso-C16:0 (1.7%), anteiso-C15:0 (1.7%), C18:1ω9c (1.7%), C16:0 1-OH (1.2%), and iso-C18:0 (1.1%). The polar lipids of the strain consisted of diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), hydroxy-phosphatidylethanolamine (OH-PE), phosphatidylinositol (PI), two phosphatidylinositol mannosides (PIMs), and an unidentified phospholipid (PL)  . All theT were affiliated with the genus Streptomyces and most closely related to S. aureoverticillatus JCM 4347T (97.9%). Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain formed a cluster with S. vastus JCM4524T (97.4%), S. cinereus DSM43033T (97.2%), S. xiangluensis NEAU-LA29T (97.1%), and S. flaveus JCM3035T (97.1%) in the neighbor-joining tree  and S. flaveus JCM 3035T (7%); and the strain could grow at pH 11.0, while S. vastus JCM4524T, S. cinereus DSM 43033T, and S. xiangluensis NEAU-LA29T could not. Other phenotypic differences included the production of H2S; decomposition of cellulose; liquefaction of gelatin; growth temperature; hydrolysis of Tweens ; and utilization of L-arabinose, D-galactose, D-fructose, D-maltose, lactose, L-rhamnose, D-ribose, D-sorbitol, D-mannose, raffinose, D-xylose, myo-inositol, L-glutamine, glycine, L-threonine, L-tyrosine, L-serine, L-proline, L-asparagine, and L-arginine.Comparison of phenotypic characteristics between strain NEAU-SSA 1 strains . DiffereT is considered to represent a novel species within the genus Streptomyces, for which the name Streptomyces sporangiiformans is proposed.On the basis of morphological, physiological, chemotaxonomic, and phylogenetic results, strain NEAU-SSA 1Streptomyces sporangiiformans .w/v) NaCl. Growth was observed at temperatures between 15 and 45 °C, with an optimum temperature of 28 °C. Positive for decomposition of Tweens (40 and 80) and cellulose, hydrolysis of aesculin and starch, liquefaction of gelatin and production of urease; and negative for coagulation and peptonization of milk, hydrolysis of Tween 20, production of H2S, and reduction of nitrate. D-fructose, D-galactose, D-glucose, inositol, lactose, D-maltose, D-mannose, D-raffinose, L-rhamnose, and D-sucrose were utilized as sole carbon sources, but not L-arabinose, dulcitol, D-ribose, D-sorbitol, or D-xylose. L-alanine, L-arginine, L-asparagine, L-aspartic acid, creatine, L-glutamic acid, L-glutamine, L-proline, L-serine, and L-threonine were utilized as sole nitrogen sources, but not glycine or L-tyrosine. Cell wall contained LL-diaminopimelic acid and the whole-cell hydrolysates were ribose, mannose, and galactose. The polar lipids contained diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), hydroxy-phosphatidylethanolamine (OH-PE), phosphatidylinositol (PI), two phosphatidylinositol mannosides (PIMs), and an unidentified phospholipid (PL). The menaquinones were MK-9(H4), MK-9(H6), and MK-9(H8). Major fatty acids were iso-C17:0, C16:0, and C17:1ω9c.Gram-stain-positive, aerobic actinomycete that formed well-developed, branched substrate hyphae and aerial mycelia. Sporangia consisted of cylindrical and rough surfaced spores (0.6–0.8 μm × 0.9–1.6 μm) were produced on aerial mycelia, but spore chains were not observed. Good growth on ISP 3, ISP 4, ISP 7, and Nutrient agar media; moderate growth on ISP 1, ISP 2, ISP 5, ISP 6, and Czapek’s agar media; and poor growth on Bennett’s agar medium. Growth occurred at pH values between 6.0 and 11.0, the optimum being pH 7.0. Tolerates up to 6.0% NaCl and grows optimally in 0–1% (T (=CCTCC AA 2017028T = DSM 105692T), isolated from soil collected from Mount Song, Dengfeng, Henan Province, China. The DNA G+C content of the type strain was 69.9 mol %, calculated from the assembly for the draft genome sequence. The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain NEAU-SSA 1T is MH842151. This Whole Genome Shotgun project has been deposited at DDBJ/ENA/GenBank under the accession VCHX00000000. The version described in this paper is version VCHX00000000.2.The type strain was NEAU-SSA 1T exhibited antibacterial activity against Ralstonia solanacearum with inhibitory zone diameters of 23 mm  suggested that strain NEAU-SSA 1T belonged to the genus Streptomyces. Physiology and biochemical characteristics, together with DDH relatedness values and ANI values, clearly indicated that strain NEAU-SSA 1T could be differentiated from the closely related strains S. aureoverticillatus JCM 4347T, S. vastus JCM 4524T, S. cinereus DSM 43033T, S. xiangluensis NEAU-LA29T, and S. flaveus JCM 3035T. Based on the polyphasic analysis, it is proposed that strain NEAU-SSA 1T should be classified as representatives of a novel species of the genus Streptomyces, for which the name Streptomyces sporangiiformans sp. nov. is proposed. The type strain is NEAU-SSA 1T (=CCTCC AA 2017028T = DSM 105692T).A novel strain NEAU-SSA 1


These articles have been corrected: The Figure 2G in ‘Upregulation of long non-coding RNA PlncRNA-1 promotes proliferation and induces epithelial-mesenchymal transition in prostate cancer,’ (https://doi.org/10.18632/oncotarget.15318), is a duplication of Figure 3F in the published article ‘PlncRNA-1 induces apoptosis through the Her-2 pathway in prostate cancer cells,’ (https://doi.org/10.4103/1008-682X.178849). The proper Figure 2G is shown below. The authors declare that these corrections do not change the results or conclusions of this paper.26090-26099.https://doi.org/10.18632/oncotarget.15318Original article: Oncotarget. 2017; 8:26090–26099.


Lysinibacillus sp. OL1 and Enterococcus sp. OL5, were isolated from boron fertilizer-amended cauliflower plantation field soils in India. Here, we report the draft genome sequences of OL1 (4.87 Mb) and OL5 (3.93 Mb) to explore the intricacies of boron tolerance in bacteria.Two novel boron-tolerant, arsenic-resistant, Gram-positive bacterial strains,  Lysinibacillus sp. OL1 and Enterococcus sp. OL5, were isolated from boron fertilizer-amended cauliflower plantation field soils in India. Here, we report the draft genome sequences of OL1 (4.87 Mb) and OL5 (3.93 Mb) to explore the intricacies of boron tolerance in bacteria.Two novel boron-tolerant, arsenic-resistant, Gram-positive bacterial strains,  The enriched culture was dilution plated on boric acid-supplemented tryptone soya agar (TSA) for isolation and screening of boron-tolerant strains, as described previously (3BO3 and sodium arsenite (NaAsO2), respectively, was tested in TSA supplemented with different concentrations of boron or arsenic , following methods described previously (Boron (B) occurs in soil solution as a nonionized molecule (Ht growth . Boron-t arsenic  with minhttp://www.usadellab.org/cms/?page=trimmomatic) high-quality data, which were subsequently assembled into 179 scaffolds  using SPAdes v3.11.1 with default parameters (http://hannonlab.cshl.edu/fastx_toolkit/download.html). For OL5, a genomic DNA library was constructed using an Ion Xpress Plus fragment library kit  and sequenced on a 530 chip of the Ion S5 system (Thermo Fisher Scientific). Before sequence retrieval from Ion S5, low-quality and polyclonal reads were filtered out and the adaptor sequences trimmed using the inbuilt PGM software. A total of 3,993,359 bp of adapter-free, quality-filtered reads  were obtained and subsequently assembled into 46 contigs  using SPAdes v3.13.0 with default parameters  under the accession numbers PRJNA521315 and PRJNA549579.These whole-genome shotgun projects have been deposited at DDBJ/ENA/GenBank under the accession numbers