Survival outcomes and interval between lymphoscintigraphy and SLNB in cutaneous melanoma- findings of a large prospective cohort study

Introduction: Sentinel lymph node biopsy (SLNB) in cutaneous melanoma (CM) is performed to identify patient at risk of regional and distant relapse. We hypothesized that timing of lymphoscintigraphy may influence the accuracy of SLNB and patient outcomes. Methods: We reviewed prospective data on patients undergoing SLNB for CM at a large university cancer-center between 2008-2015, examining patient and tumor demographics and time between lymphoscintigraphy (LS) and SLNB. Kaplan-Meier survival analysis assessed disease-specific (DSS) and overall-survival (OS), stratified by timing of LS. Cox multivariate regression analysis assessed independent risk factors for survival. Results: We identified 1015 patients. Median follow-up was 45 months (IQR 26-68 months). Univariate analysis showed a 6.8% absolute DSS (HR 1.6 [1.03-2.48], p= 0.04) benefit and a 10.7% absolute OS (HR 1.64 [1.13-2.38], p=0.01) benefit for patients whose SLNB was performed 12 hours (n=652). Multivariate analysis identified timing of LS as an independent predictor of OS (p=0.007) and DSS (p=0.016) when competing with age, sex, Breslow thickness (BT) and SLN status. No difference in nodal relapse rates (5.2% v 4.6%; p=0.67) was seen. Both groups were matched for age, sex, BT and SLN status. Conclusion: These data have significant implications for SLNB services, suggesting delaying SLNB >12 hours after LS using a Tc99-labelled nanocolloid has a significant negative survival impact for patients and should be avoided. We hypothesise that temporal tracer migration is the underlying cause and advocate further trials investigating alternative, 'stable' tracer-agents

A feasibility study of indocyanine green fluorescence mapping for sentinel lymph node detection in cutaneous melanoma

Objectives: Sentinel lymph node biopsy (SLNB) is standard of care for staging regional LN in AJCC stage IB-IIC melanoma; using dual localization with radiolabelled colloid and blue dye. Combining these gives optimal accuracy; drawbacks include cumulative radiation exposure for healthcare workers, coordination between disciplines and anaphylaxis. An alternative tracer agent is indocyanine green (ICG); an optical enhancer that fluoresces in the near infrared range. This prospective cohort study assesses the feasibility of using ICG as a tracer agent to detect SLN in cutaneous melanoma. Methods: Primary melanoma patients diagnosed with pT1b-pT4b tumours undergoing SLNB were recruited over a 6-month period at a tertiary referral centre. All underwent standard preoperative lymphoscintigraphy (LSG) using 20-40MBq of Tc99radiolabelled nanocolloid plus intraoperative Patent Blue dye (PBD). ICG was administered as a third tracer agent intraoperatively. Results: 62 patients (33M/29F) were recruited; median age was 61 years. Median melanoma Breslow thickness was 1.6mm. 144 specimens containing 135 SLN were excised. Concordance rate for all 3 tracer agents was 88.1%(119/135 LN); that for radioisotope/PBD was 88.2%(95%CI:82.2,93.7). There were no discordance pairs between radioisotope/PBD compared to radioisotope/PBD/ICG. Radioisotope/ICG significantly increased the sensitivity of detecting SLN to 98.5%(95%CI:94.8,99.8); p<0.00001 compared to radioisotope/PBD. Concordance rate of intraoperative ICG drainage pattern with LSG was 22.6%. Conclusion: ICG utilization showed comparable sensitivity with gold standard. Technical challenges e.g. ICG leakage into biopsy field, poor concordance with LSG limits its efficacy in melanoma SLNB. We therefore do not recommend replacing current practice with ICG alone or by using a combination with TC99

The oxygen isotopic composition of phosphate in river water and its potential sources in the Upper River Taw catchment, UK

The need to reduce both point and diffuse phosphorus pollution to aquatic ecosystems is widely recognised and in order to achieve this, identification of the different pollutant sources is essential. Recently, a stable isotope approach using oxygen isotopes within phosphate (δ18OPO4) has been used in phosphorus source tracing studies. This approach was applied in a one-off survey in September 2013 to the River Taw catchment in south-west England where elevated levels of phosphate have been reported. River water δ18OPO4 along the main channel varied little, ranging from + 17.1 to + 18.8‰. This was no > 0.3‰ different to that of the isotopic equilibrium with water (Eδ18OPO4). The δ18OPO4 in the tributaries was more variable (+ 17.1 to + 18.8‰), but only deviated from Eδ18OPO4 by between 0.4 and 0.9‰. Several potential phosphate sources within the catchment were sampled and most had a narrow range of δ18OPO4 values similar to that of river Eδ18OPO4. Discharge from two waste water treatment plants had different and distinct δ18OPO4 from one another ranging between + 16.4 and + 19.6‰ and similar values to that of a dairy factory final effluent (+ 16.5 to + 17.8‰), mains tap water (+ 17.8 to + 18.4‰), and that of the phosphate extracted from river channel bed sediment (+ 16.7 to + 17.6‰). Inorganic fertilizers had a wide range of values (+ 13.3 to + 25.9‰) while stored animal wastes were consistently lower (+ 12.0 to + 15.0‰) than most other sources and Eδ18OPO4. The distinct signals from the waste water treatment plants were lost within the river over a short distance suggesting that rapid microbial cycling of phosphate was occurring, because microbial cycling shifts the isotopic signal towards Eδ18OPO4. This study has added to the global inventory of phosphate source δ18OPO4 values, but also demonstrated the limitations of this approach to identifying phosphate sources, especially at times when microbial cycling is high

Souporcell: robust clustering of single-cell RNA-seq data by genotype without reference genotypes.

Methods to deconvolve single-cell RNA-sequencing (scRNA-seq) data are necessary for samples containing a mixture of genotypes, whether they are natural or experimentally combined. Multiplexing across donors is a popular experimental design that can avoid batch effects, reduce costs and improve doublet detection. By using variants detected in scRNA-seq reads, it is possible to assign cells to their donor of origin and identify cross-genotype doublets that may have highly similar transcriptional profiles, precluding detection by transcriptional profile. More subtle cross-genotype variant contamination can be used to estimate the amount of ambient RNA. Ambient RNA is caused by cell lysis before droplet partitioning and is an important confounder of scRNA-seq analysis. Here we develop souporcell, a method to cluster cells using the genetic variants detected within the scRNA-seq reads. We show that it achieves high accuracy on genotype clustering, doublet detection and ambient RNA estimation, as demonstrated across a range of challenging scenarios.We acknowledge the Wellcome Sanger Institute’s DNA Pipelines for construction of the 10x sequencing libraries. We thank Allan Muhwezi and Andrew Russell for assistance with parasite culture and 10x Single-cell 3’ RNA-seq respectively. In addition, we would like to thank Matthew Young for useful conversations about ambient RNA, Mirjana Efremova for providing information about the maternal/fetal data, and Katie Gray for assistance in interpreting the previously unannotated cluster. The Wellcome Sanger Institute is funded by the Wellcome Trust (grant 206194/Z/17/Z), which supports MKNL and MH. This work was supported by an MRC Career Development Award (G1100339) to MKNL. We would like to acknowledge the Wellcome Trust Sanger Institute as the source of the human induced pluripotent cell lines that were generated under the Human Induced Pluripotent Stem Cell Initiative funded by a grant from the Wellcome Trust and Medical Research Council, supported by the Wellcome Trust (WT098051) and the NIHR/Wellcome Trust Clinical Research Facility, and acknowledges Life Science Technologies Corporation as the provider of Cytotune (HipSci.org). The Cardiovascular Epidemiology Unit is supported by core funding from the UK Medical Research Council (MR/L003120/1), the British Heart Foundation (RG/13/13/30194; RG/18/13/33946) and the National Institute for Health Research [Cambridge Biomedical Research Centre at the Cambridge University Hospital’s NHS Foundation Trust]. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care

Improved perioperative seroma and complication rates following the application of a 2-layer negative pressure wound therapy system after inguinal lymphadenectomy for metastatic cutaneous melanoma

Background: Perioperative complications following inguinal lymphadenectomy, including seroma formation, are frequent. We have employed a 2-layer negative pressure wound therapy (2-LNPWT) as a method to reduce seroma rate and perioperative complications. We present the outcome of our initial experience with 2-LNPWT and compare the outcomes of its use with traditional closed suction drains (CSDs).  Materials and methods: A non-randomised retrospective case–control series was analysed. Surgeons performing inguinal lymphadenectomy for metastatic cutaneous melanoma utilised either the 2-LNPWT therapy or traditional CSDs according to their practice preference.  Results: The study included 111 patients. The cohorts were well matched for gender, disease burden, body mass index and comorbidities. The 2-LNPWT technique was associated with significantly better postoperative outcomes than CSD, in terms of incidence of seroma formation (26.9% vs 49.4%; p < 0.03), period of drainage (15 days vs 20 days; p = 0.005) and return to theatre rate (0% vs 15.3%; p = 0.03). The overall seroma rate was 44.1%. The only significant association with seroma initiation was the type of drainage system used (2-LNPWT 31.2% vs CSD 58.3%; p < 0.03; OR 3.0). The method of drainage did not alter the course of an established seroma. There was no significant difference in overall or disease-specific survival detected between the 2 groups.  Conclusion: This retrospective non-randomised case control study has demonstrated the safe use of a novel application of negative pressure wound therapy that significantly reduced the incidence of seroma formation and postoperative complication rate for inguinal lymphadenectomy for melanoma

Observations of Massive Star Forming Regions with Water Masers: Mid-Infrared Imaging

We present here a mid-infrared imaging survey of 26 sites of water maser emission. Observations were obtained at the InfraRed Telescope Facility 3-m telescope with the University of Florida mid-infrared imager/spectrometer OSCIR, and the JPL mid-infrared camera MIRLIN. The main purpose of the survey was to explore the relationship between water masers and the massive star formation process. It is generally believed that water masers predominantly trace outflows and embedded massive stellar objects, but may also exist in circumstellar disks around young stars. We investigate each of these possibilities in light of our mid-infrared imaging. We find that mid-infrared emission seems to be more closely associated with water and OH maser emission than cm radio continuum emission from UC HII regions. We also find from the sample of sources in our survey that, like groups of methanol masers, both water and OH masers have a proclivity for grouping into linear or elongated distributions. We conclude that the vast majority of linearly distributed masers are not tracing circumstellar disks, but outflows and shocks instead.Comment: 49 pages; 23 figures; To appear in February 2005 ApJS; To download a version with better quality figures, go to http://www.ctio.noao.edu/~debuizer

Risk Stratification of Sentinel Node Metastasis Disease Burden and Phenotype in Stage III Melanoma Patients

Background: Currently, all patients with American Joint Committee on Cancer (AJCC) pT2b-pT4b melanomas and a positive sentinel node biopsy are now considered for adjuvant systemic therapy without consideration of the burden of disease in the metastatic nodes. Methods: This was a retrospective cohort analysis of 1377 pT1-pT4b melanoma patients treated at an academic cancer center. Standard variables regarding patient, primary tumor, and sentinel node characteristics, in addition to sentinel node metastasis maximum tumor deposit size (MTDS) in millimeters and extracapsular spread (ECS) status, were analyzed for predicting disease-specific survival (DSS). Results: The incidence of SN+ was 17.3% (238/1377) and ECS was 10.5% (25/238). Increasing AJCC N stage was associated with worse DSS. There was no difference in DSS between the IIIB and IIIC groups. Subgroup analyses showed that the optimal MTDS cut-point was 0.7 mm for the pT1b-pT4a SN+ subgroups, but there was no cut-point for the pT4b SN+ subgroup. Patients with MTDS <0.7 mm and no ECS had similar survival outcomes as the N0 patients with the same T stage. Nodal risk categories were developed using the 0.7 mm MTDS cut-point and ECS status. The incidence of low-risk disease, according to the new nodal risk model, was 22.3% (53/238) in the stage III cohort, with 49% (26/53) in the pT2b-pT3a and pT3b-pT4a subgroups and none in the pT4b subgroup. Similar outcomes were observed for overall and distant metastasis-free survival. Conclusion: We propose a more granular classification system, based on tumor burden and ECS status in the sentinel node, that identifies low-risk patients in the AJCC IIIB and IIIC subgroups who may otherwise be observed

Neurocognitive function in HIV infected patients on antiretroviral therapy

OBJECTIVE To describe factors associated with neurocognitive (NC) function in HIV-positive patients on stable combination antiretroviral therapy. DESIGN We undertook a cross-sectional analysis assessing NC data obtained at baseline in patients entering the Protease-Inhibitor-Monotherapy-Versus-Ongoing-Triple therapy (PIVOT) trial. MAIN OUTCOME MEASURE NC testing comprised of 5 domains. Raw results were z-transformed using standard and demographically adjusted normative datasets (ND). Global z-scores (NPZ-5) were derived from averaging the 5 domains and percentage of subjects with test scores >1 standard deviation (SD) below population means in at least two domains (abnormal Frascati score) calculated. Patient characteristics associated with NC results were assessed using multivariable linear regression. RESULTS Of the 587 patients in PIVOT, 557 had full NC results and were included. 77% were male, 68% Caucasian and 28% of Black ethnicity. Mean (SD) baseline and nadir CD4+ lymphocyte counts were 553(217) and 177(117) cells/µL, respectively, and HIV RNA was <50 copies/mL in all. Median (IQR) NPZ-5 score was -0.5 (-1.2/-0) overall, and -0.3 (-0.7/0.1) and -1.4 (-2/-0.8) in subjects of Caucasian and Black ethnicity, respectively. Abnormal Frascati scores using the standard-ND were observed in 51%, 38%, and 81%, respectively, of subjects overall, Caucasian and Black ethnicity (p<0.001), but in 62% and 69% of Caucasian and Black subjects using demographically adjusted-ND (p = 0.20). In the multivariate analysis, only Black ethnicity was associated with poorer NPZ-5 scores (P<0.001). CONCLUSIONS In this large group of HIV-infected subjects with viral load suppression, ethnicity but not HIV-disease factors is closely associated with NC results. The prevalence of abnormal results is highly dependent on control datasets utilised. TRIAL REGISTRY ClinicalTrials.gov, NCT01230580

Marine20—the marine radiocarbon age calibration curve (0 – 55,000 cal BP)

T.J. Heaton is supported by a Leverhulme Trust Fellowship RF-2019-140\9, “Improving the Measurement of Time Using Radiocarbon”. M Butzin is supported by the German Federal Ministry of Education and Research (BMBF), as Research for Sustainability initiative (FONA); www.fona.de through the PalMod project (grant numbers: 01LP1505B, 01LP1919A). E. Bard is supported by EQUIPEX ASTER-CEREGE and ANR CARBOTRYDH. Meetings of the IntCal Marine Focus group have been supported by Collège de France.The concentration of radiocarbon (14C) differs between ocean and atmosphere. Radiocarbon determinations from samples which obtained their 14C in the marine environment therefore need a marine-specific calibration curve and cannot be calibrated directly against the atmospheric-based IntCal20 curve. This paper presents Marine20, an update to the internationally agreed marine radiocarbon age calibration curve that provides a non-polar global-average marine record of radiocarbon from 0–55 cal kBP and serves as a baseline for regional oceanic variation. Marine20 is intended for calibration of marine radiocarbon samples from non-polar regions; it is not suitable for calibration in polar regions where variability in sea ice extent, ocean upwelling and air-sea gas exchange may have caused larger changes to concentrations of marine radiocarbon. The Marine20 curve is based upon 500 simulations with an ocean/atmosphere/biosphere box-model of the global carbon cycle that has been forced by posterior realizations of our Northern Hemispheric atmospheric IntCal20 14C curve and reconstructed changes in CO2 obtained from ice core data. These forcings enable us to incorporate carbon cycle dynamics and temporal changes in the atmospheric 14C level. The box-model simulations of the global-average marine radiocarbon reservoir age are similar to those of a more complex three-dimensional ocean general circulation model. However, simplicity and speed of the box model allow us to use a Monte Carlo approach to rigorously propagate the uncertainty in both the historic concentration of atmospheric 14C and other key parameters of the carbon cycle through to our final Marine20 calibration curve. This robust propagation of uncertainty is fundamental to providing reliable precision for the radiocarbon age calibration of marine based samples. We make a first step towards deconvolving the contributions of different processes to the total uncertainty; discuss the main differences of Marine20 from the previous age calibration curve Marine13; and identify the limitations of our approach together with key areas for further work. The updated values for ΔR, the regional marine radiocarbon reservoir age corrections required to calibrate against Marine20, can be found at the data base http://calib.org/marine/.Publisher PDFPeer reviewe