Residual ctDNA after treatment predicts early relapse in patients with early-stage non-small cell lung cancer.

BACKGROUND: Identification of residual disease in patients with localized non-small cell lung cancer (NSCLC) following treatment with curative intent holds promise to identify patients at risk of relapse. New methods can detect circulating tumour DNA (ctDNA) in plasma to fractional concentrations as low as a few parts per million, and clinical evidence is required to inform their use. PATIENTS AND METHODS: We analyzed 363 serial plasma samples from 88 patients with early-stage NSCLC (48.9%/28.4%/22.7% at stage I/II/III), predominantly adenocarcinomas (62.5%), treated with curative intent by surgery (n = 61), surgery and adjuvant chemotherapy/radiotherapy (n = 8), or chemoradiotherapy (n = 19). Tumour exome sequencing identified somatic mutations and plasma was analyzed using patient-specific RaDaR™ assays with up to 48 amplicons targeting tumour-specific variants unique to each patient. RESULTS: ctDNA was detected before treatment in 24%, 77% and 87% of patients with stage I, II and III disease, respectively, and in 26% of all longitudinal samples. The median tumour fraction detected was 0.042%, with 63% of samples <0.1% and 36% of samples <0.01%. ctDNA detection had clinical specificity >98.5% and preceded clinical detection of recurrence of the primary tumour by a median of 212.5 days. ctDNA was detected after treatment in 18/28 (64.3%) of patients who had clinical recurrence of their primary tumour. Detection within the landmark timepoint 2 weeks to 4 months after treatment end occurred in 17% of patients, and was associated with shorter recurrence-free survival [hazard ratio (HR): 14.8, P <0.00001] and overall survival (HR: 5.48, P <0.0003). ctDNA was detected 1-3 days after surgery in 25% of patients yet was not associated with disease recurrence. Detection before treatment was associated with shorter overall survival and recurrence-free survival (HR: 2.97 and 3.14, P values 0.01 and 0.003, respectively). CONCLUSIONS: ctDNA detection after initial treatment of patients with early-stage NSCLC using sensitive patient-specific assays has potential to identify patients who may benefit from further therapeutic intervention.

Infant Effortful Control Mediates Relations Between Nondirective Parenting and Internalising-Related Child Behaviours in an Autism-Enriched Infant Cohort.

Internalising problems are common within Autism Spectrum Disorder (ASD); early intervention to support those with emerging signs may be warranted. One promising signal lies in how individual differences in temperament are shaped by parenting. Our longitudinal study of infants with and without an older sibling with ASD investigated how parenting associates with infant behavioural inhibition (8-14 months) and later effortful control (24 months) in relation to 3-year internalising symptoms. Mediation analyses suggest nondirective parenting (8 months) was related to fewer internalising problems through an increase in effortful control. Parenting did not moderate the stable predictive relation of behavioural inhibition on later internalising. We discuss the potential for parenting to strengthen protective factors against internalising in infants from an ASD-enriched cohort.

Anxious parents show higher physiological synchrony with their infants.

BACKGROUND: Interpersonal processes influence our physiological states and associated affect. Physiological arousal dysregulation, a core feature of anxiety disorders, has been identified in children of parents with elevated anxiety. However, little is understood about how parent-infant interpersonal regulatory processes differ when the dyad includes a more anxious parent. METHODS: We investigated moment-to-moment fluctuations in arousal within parent-infant dyads using miniaturised microphones and autonomic monitors. We continually recorded arousal and vocalisations in infants and parents in naturalistic home settings across day-long data segments. RESULTS: Our results indicated that physiological synchrony across the day was stronger in dyads including more rather than less anxious mothers. Across the whole recording epoch, less anxious mothers showed responsivity that was limited to 'peak' moments in their child's arousal. In contrast, more anxious mothers showed greater reactivity to small-scale fluctuations. Less anxious mothers also showed behaviours akin to 'stress buffering' - downregulating their arousal when the overall arousal level of the dyad was high. These behaviours were absent in more anxious mothers. CONCLUSION: Our findings have implications for understanding the differential processes of physiological co-regulation in partnerships where a partner is anxious, and for the use of this understanding in informing intervention strategies for dyads needing support for elevated levels of anxiety.

Gulf of Mexico blue hole harbors high levels of novel microbial lineages.

Exploration of oxygen-depleted marine environments has consistently revealed novel microbial taxa and metabolic capabilities that expand our understanding of microbial evolution and ecology. Marine blue holes are shallow karst formations characterized by low oxygen and high organic matter content. They are logistically challenging to sample, and thus our understanding of their biogeochemistry and microbial ecology is limited. We present a metagenomic and geochemical characterization of Amberjack Hole on the Florida continental shelf (Gulf of Mexico). Dissolved oxygen became depleted at the hole's rim (32 m water depth), remained low but detectable in an intermediate hypoxic zone (40-75 m), and then increased to a secondary peak before falling below detection in the bottom layer (80-110 m), concomitant with increases in nutrients, dissolved iron, and a series of sequentially more reduced sulfur species. Microbial communities in the bottom layer contained heretofore undocumented levels of the recently discovered phylum Woesearchaeota (up to 58% of the community), along with lineages in the bacterial Candidate Phyla Radiation (CPR). Thirty-one high-quality metagenome-assembled genomes (MAGs) showed extensive biochemical capabilities for sulfur and nitrogen cycling, as well as for resisting and respiring arsenic. One uncharacterized gene associated with a CPR lineage differentiated hypoxic from anoxic zone communities. Overall, microbial communities and geochemical profiles were stable across two sampling dates in the spring and fall of 2019. The blue hole habitat is a natural marine laboratory that provides opportunities for sampling taxa with under-characterized but potentially important roles in redox-stratified microbial processes.

Experimental Realization of a Quantum Dot Energy Harvester.

We demonstrate experimentally an autonomous nanoscale energy harvester that utilizes the physics of resonant tunneling quantum dots. Gate-defined quantum dots on GaAs/AlGaAs high-electron-mobility transistors are placed on either side of a hot-electron reservoir. The discrete energy levels of the quantum dots are tuned to be aligned with low energy electrons on one side and high energy electrons on the other side of the hot reservoir. The quantum dots thus act as energy filters and allow for the conversion of heat from the cavity into electrical power. Our energy harvester, measured at an estimated base temperature of 75 mK in a He^{3}/He^{4} dilution refrigerator, can generate a thermal power of 0.13 fW for a temperature difference across each dot of about 67 mK.

Why is the H3+ hot spot above Jupiter's Great Red Spot so hot?

Recent observations of Jupiter's Great Red Spot indicate that the thermosphere above the storm is hotter than its surroundings by more than 700 K. Possible suggested sources for this heating have thus far included atmospheric gravity waves and lightning-driven acoustic waves. Here, we propose that Joule heating, driven by Great Red Spot vorticity penetrating up into the lower stratosphere and coupling to the thermosphere, may contribute to the large observed temperatures. The strength of Joule heating will depend on the local inclination angle of the magnetic field and thus the observed emissions and inferred temperatures should vary with planetary longitude as the Great Red Spot tracks across the planet. This article is part of a discussion meeting issue 'Advances in hydrogen molecular ions: H3+, H5+ and beyond'.

Effects of late gestation shearing on BW, feed intake and plasma metabolite concentrations in Rambouillet ewes managed outdoors during winter.

The majority of lambs in the United States are born from late winter to early spring and pregnant ewes are generally sheared in the last third of pregnancy. Although there are benefits to shearing before parturition, shorn animals may be more vulnerable to the cold, highly variable climatic conditions associated with these seasons. The objective of this study was to determine if late gestation shearing induces differences in individual BW, dry matter intake (DMI) and plasma metabolite concentration of finewool ewes managed outdoors during winter. Thirty-six mature, pregnant Rambouillet ewes (3.8±0.45 years; 76.8±11.4 kg) were managed in a drylot with ad libitum access to pelleted alfalfa in bunks capable of measuring individual daily DMI. The treatment group consisted of ewes sheared at ~5 weeks before the estimated parturition date (shorn; n=18). Unshorn ewes (n=18) remained in full fleece throughout the experiment and were shorn on the last day of the experiment ~2 weeks before the estimated parturition date. Blood was collected on days 0 (before shearing shorn group), 7, 14 and 21 (before shearing unshorn group) of the trial, and plasma was isolated and analyzed for non-esterified fatty acid (NEFA), ß-hydroxybutyrate (BHB) and glucose (GLU) concentrations. There was no effect of shearing on ewe DMI or BW during the trial (P⩾0.35). Plasma NEFA and GLU concentrations were similar (P⩾0.36) between shearing groups, though plasma BHB concentration was 103.7 µmol/l greater (24.1%; P<0.01) in unshorn ewes. Lamb BW at birth was not affected (P=0.30) by ewe shearing treatment. Under conditions of this study, no differences in economically important aspects of sheep production were observed between shorn and unshorn pregnant ewes.

Association Of Plasma And Urinary Mutant DNA With Clinical Outcomes In Muscle Invasive Bladder Cancer.

Muscle Invasive Bladder Cancer (MIBC) has a poor prognosis. Whilst patients can achieve a 6% improvement in overall survival with Neo-Adjuvant Chemotherapy (NAC), many do not respond. Body fluid mutant DNA (mutDNA) may allow non-invasive identification of treatment failure. We collected 248 liquid biopsy samples including plasma, cell pellet (UCP) and supernatant (USN) from spun urine, from 17 patients undergoing NAC. We assessed single nucleotide variants and copy number alterations in mutDNA using Tagged-Amplicon- and shallow Whole Genome- Sequencing. MutDNA was detected in 35.3%, 47.1% and 52.9% of pre-NAC plasma, UCP and USN samples respectively, and urine samples contained higher levels of mutDNA (p = <0.001). Longitudinal mutDNA demonstrated tumour evolution under the selective pressure of NAC e.g. in one case, urine analysis tracked two distinct clones with contrasting treatment sensitivity. Of note, persistence of mutDNA detection during NAC predicted disease recurrence (p = 0.003), emphasising its potential as an early biomarker for chemotherapy response.

Investigating the poor outcomes of BRAF-mutant advanced colorectal cancer: analysis from 2530 patients in randomised clinical trials.

Background: To improve strategies for the treatment of BRAF-mutant advanced colorectal cancer (aCRC) patients, we examined individual data from patients treated with chemotherapy alone in three randomised trials to identify points on the treatment pathway where outcomes differ from BRAF wild-types. Patients and methods: 2530 aCRC patients were assessed from three randomised trials. End-points were progression-free survival, response rate, disease control rate, post-progression survival (P-PS) and overall survival. Treatments included first-line oxaliplatin/fluorouracil (OxFU) and second-line irinotecan. Clinicians were unaware of BRAF-status. Results: 231 patients (9.1%) had BRAF-mutant tumours. BRAF-mutation conferred significantly worse survival independent of associated clinicopathological factors known to be prognostic. Compared with wild-type, BRAF-mutant patients treated with first-line OxFU had similar DCR (59.2% versus 72%; adjusted OR = 0.76, P = 0.24) and PFS (5.7 versus 6.3 months; adjusted HR = 1.14, P = 0.26). Following progression on first-line chemotherapy, BRAF-mutant patients had a markedly shorter P-PS (4.2 versus 9.2 months, adjusted HR = 1.69, P < 0.001). Fewer BRAF-mutant patients received second-line treatment (33% versus 51%, P < 0.001), but BRAF-mutation was not associated with inferior second-line outcomes (RR adjusted OR = 0.56, P = 0.45; PFS adjusted HR = 1.01, P = 0.93). Significant clinical heterogeneity within the BRAF-mutant population was observed: a proportion (24.3%) had good first-line PFS and P-PS (both >6 months; OS = 24.0 months); however, 36.5% progressed rapidly through first-line chemotherapy and thereafter, with OS = 4.7 months. Conclusions: BRAF-mutant aCRC confers a markedly worse prognosis independent of associated clinicopathological features. Chemotherapy provides meaningful improvements in outcome throughout treatment lines. Post-progression survival is markedly worse and vigilance is required to ensure appropriate delivery of treatment after first-line progression.

Advanced maternal age causes adverse programming of mouse blastocysts leading to altered growth and impaired cardiometabolic health in post-natal life.

STUDY QUESTION: Does advanced maternal age (AMA) in mice affect cardiometabolic health during post-natal life in offspring derived from an assisted reproduction technology (ART) procedure? SUMMARY ANSWER: Offspring derived from blastocysts collected from aged female mice displayed impaired body weight gain, blood pressure, glucose metabolism and organ allometry during post-natal life compared with offspring derived from blastocysts from young females; since all blastocysts were transferred to normalized young mothers, this effect is independent of maternal pregnancy conditions. WHAT IS KNOWN ALREADY: Although studies in mice have shown that AMA can affect body weight and behaviour of offspring derived from natural reproduction, data on the effects of AMA on offspring cardiometabolic health during post-natal development are not available. Given the increasing use of ART to alleviate infertility in women of AMA, it is pivotal to develop ART-AMA models addressing the effects of maternal aging on offspring health. STUDY DESIGN, SIZE, DURATION: Blastocysts from old (34-39 weeks) or young (8-9 weeks) C57BL/6 females mated with young CBA males (13-15 weeks) were either subjected to differential cell staining (inner cell mass and trophectoderm) or underwent embryo transfer (ET) into young MF1 surrogates (8-9 weeks) to produce young (Young-ET, 9 litters) and old (Old-ET, 10 litters) embryo-derived offspring. Offspring health monitoring was carried out for 30 weeks. PARTICIPANTS/MATERIALS, SETTING, METHODS: All animals were fed with standard chow. Blood pressure was measured at post-natal Weeks 9, 15 and 21, and at post-natal Week 30 a glucose tolerance test (GTT) was performed. Two days after the GTT mice were killed for organ allometry. Blastocyst cell allocation variables were evaluated by T-test and developmental data were analysed with a multilevel random effects regression model. MAIN RESULTS AND THE ROLE OF CHANCE: The total number of cells in blastocysts from aged mice was decreased (P < 0.05) relative to young mice due to a lower number of cells in the trophectoderm (mean ± SEM: 34.5 ± 2.1 versus 29.6 ± 1.0). Weekly body weight did not differ in male offspring, but an increase in body weight from Week 13 onwards was observed in Old-ET females (final body weight at post-natal Week 30: 38.5 ± 0.8 versus 33.4 ± 0.8 g, P < 0.05). Blood pressure was increased in Old-ET offspring at Weeks 9-15 in males (Week 9: 108.5 ± 3.13 versus 100.8 ± 1.5 mmHg, Week 15: 112.9 ± 3.2 versus 103.4 ± 2.1 mmHg) and Week 15 in females (115.9 ± 3.7 versus 102.8 ± 0.7 mmHg; all P < 0.05 versus Young-ET). The GTT results and organ allometry were not affected in male offspring. In contrast, Old-ET females displayed a greater (P < 0.05) peak glucose concentration at 30 min during the GTT (21.1 ± 0.4 versus 17.8 ± 1.16 mmol/l) and their spleen weight (88.2 ± 2.6 ± 105.1 ± 4.6 mg) and several organ:body weight ratios (g/g × 10(3)) were decreased (P < 0.05 versus Young-ET), including the heart (3.7 ± 0.06 versus 4.4 ± 0.08), lungs (4.4 ± 0.1 versus 5.0 ± 0.1), spleen (2.4 ± 0.06 versus 3.2 ± 0.1) and liver (36.4 ± 0.6 versus 39.1 ± 0.9). LIMITATIONS, REASONS FOR CAUTION: Results from experimental animal models cannot be extrapolated to humans. Nevertheless, they are valuable to develop conceptual models that can produce hypotheses for eventual testing in the target species (i.e. humans). WIDER IMPLICATIONS OF THE FINDINGS: Our data show that offspring from mouse embryos from aged mothers can develop altered phenotypes during post-natal development compared with embryos from young mothers. Because all embryos were transferred into young mothers for the duration of pregnancy to normalize the maternal in vivo environment, our findings indicate that adverse programming via AMA is already established at the blastocyst stage. Whilst human embryos display increased aneuploidy compared with mouse, we believe our data have implications for women of AMA undergoing assisted reproduction, including surrogacy programmes. STUDY FUNDING/COMPETING INTERESTS: This work was supported through the European Union FP7-CP-FP Epihealth programme (278418) to T.P.F. and the BBSRC (BB/F007450/1) to T.P.F. The authors have no conflicts of interest to declare.

Review of brucellosis cases from laboratory exposures in the United States in 2008 to 2011 and improved strategies for disease prevention.

Five laboratory-acquired brucellosis (LAB) cases that occurred in the United States between 2008 and 2011 are presented. The Centers for Disease Control and Prevention (CDC) reviewed the recommendations published in 2008 and the published literature to identify strategies to further prevent LAB. The improved prevention strategies are described.

Gigahertz quantized charge pumping in graphene quantum dots.

Single-electron pumps are set to revolutionize electrical metrology by enabling the ampere to be redefined in terms of the elementary charge of an electron. Pumps based on lithographically fixed tunnel barriers in mesoscopic metallic systems and normal/superconducting hybrid turnstiles can reach very small error rates, but only at megahertz pumping speeds that correspond to small currents of the order of picoamperes. Tunable barrier pumps in semiconductor structures are operated at gigahertz frequencies, but the theoretical treatment of the error rate is more complex and only approximate predictions are available. Here, we present a monolithic, fixed-barrier single-electron pump made entirely from graphene that performs at frequencies up to several gigahertz. Combined with the record-high accuracy of the quantum Hall effect and proximity-induced Josephson junctions, quantized-current generation brings an all-graphene closure of the quantum metrological triangle within reach. Envisaged applications for graphene charge pumps outside quantum metrology include single-photon generation via electron-hole recombination in electrostatically doped bilayer graphene reservoirs, single Dirac fermion emission in relativistic electron quantum optics and read-out of spin-based graphene qubits in quantum information processing.

Non-ohmic behavior of carrier transport in highly disordered graphene.

We report measurements of disordered graphene probed by both a high electric field and a high magnetic field. By applying a high source-drain voltage, Vsd, we are able to study the current-voltage relation I-Vsd of our device. With increasing Vsd, a crossover from the linear I-Vsd regime to the non-linear one, and eventually to activationless-hopping transport occurs. In the activationless-hopping regime, the importance of Coulomb interactions between charged carriers is demonstrated. Moreover, we show that delocalization of carriers which are strongly localized at low T and at small Vsd occurs in the presence of high electric field and perpendicular magnetic field.

Tunable Kondo physics in a carbon nanotube double quantum dot.

We investigate a tunable two-impurity Kondo system in a strongly correlated carbon nanotube double quantum dot, accessing the full range of charge regimes. In the regime where both dots contain an unpaired electron, the system approaches the two-impurity Kondo model. At zero magnetic field the interdot coupling disrupts the Kondo physics and a local singlet state arises, but we are able to tune the crossover to a Kondo screened phase by application of a magnetic field. All results show good agreement with a numerical renormalization group study of the device.

Unraveling quantum Hall breakdown in bilayer graphene with scanning gate microscopy.

Investigating the structure of quantized plateaus in the Hall conductance of graphene is a powerful way of probing its crystalline and electronic structure and will also help to establish whether graphene can be used as a robust standard of resistance for quantum metrology. We use low-temperature scanning gate microscopy to image the interplateau breakdown of the quantum Hall effect in an exfoliated bilayer graphene flake. Scanning gate images captured during breakdown exhibit intricate patterns where the conductance is strongly affected by the presence of the scanning probe tip. The maximum density and intensity of the tip-induced conductance perturbations occur at half-integer filling factors, midway between consecutive quantum Hall plateau, while the intensity of individual sites shows a strong dependence on tip-voltage. Our results are well-described by a model based on quantum percolation which relates the points of high responsivity to tip-induced scattering in a network of saddle points separating localized states.

Measuring the complex admittance of a carbon nanotube double quantum dot.

We investigate radio-frequency (rf) reflectometry in a tunable carbon nanotube double quantum dot coupled to a resonant circuit. By measuring the in-phase and quadrature components of the reflected rf signal, we are able to determine the complex admittance of the double quantum dot as a function of the energies of the single-electron states. The measurements are found to be in good agreement with a theoretical model of the device in the incoherent limit. In addition to being of fundamental interest, our results present an important step forward towards noninvasive charge and spin state readout in carbon nanotube quantum dots.

Transport spectroscopy of an impurity spin in a carbon nanotube double quantum dot.

We make use of spin selection rules to investigate the electron spin system of a carbon nanotube double quantum dot. Measurements of the electron transport as a function of the magnetic field and energy detuning between the quantum dots reveal an intricate pattern of the spin state evolution. We demonstrate that the complete set of measurements can be understood by taking into account the interplay between spin-orbit interaction and a single impurity spin coupled to the double dot. The detection and tunability of this coupling are important for quantum manipulation in carbon nanotubes.

Nanoanalysis of graphene layers using scanning probe techniques.

Graphene is an almost ideal two-dimensional system. Unlike other two-dimensional electron gas systems realized in silicon or gallium arsenide, the electron wave functions are very close to the surrounding environment. While this causes problems in trying to passivate the surface without reducing the mobility, it does allow direct electrical access to the two-dimensional surface states using scanning probe techniques. In this review, we look at recent advances in the nanoanalytics of the surface and edges of graphene using scanning probe techniques.