Methodological development and biological observations of cell free DNA with a simple direct fluorescent assay in colorectal cancer.

BACKGROUND: Cell free DNA (cfDNA) has shown promising utility as prognostic biomarker for patients with colorectal cancer (CRC), with an ongoing need to optimize and validate the laboratory methodology. Here, we report our optimization and validation of a direct fluorescent assay and display the potential utility in patients with colorectal cancer. METHODS: Plasma cfDNA was analyzed by a direct fluorescent assay (DFA) and compared to quantification by droplet digital PCR (ddPCR). For clinical validation, baseline blood samples were available for a total of 273 patients from six different Nordic trials, covering patients with locally advanced rectal cancer (n = 176, cohorts A + B), liver limited metastatic CRC (n = 75C + D) and wide spread metastatic CRC (n = 22 E + F). RESULTS: Validating the DFA analysis with ddPCR revealed a strong correlation with an R2 of 0.81. For the clinical cohorts, the levels of cfDNA were: 0.8 ng/uL (95%CI 0.75-0.83) (A + B), 0.93 ng/uL (95%CI 0.86-1.02) (C + D) and 1.2 ng/uL (95%CI 0.85-1.47) (E + F), respectively (p < 0.01). All cohorts of colorectal cancer had higher levels of cell free DNA than healthy individuals (n = 94) (p < 0.01). CONCLUSION: Analysis of cell free DNA by a direct fluorescent assay could be an attractive laboratory option for a rapid inexpensive quantification of cell free DNA.

On-Orbit Performance of the Helioseismic and Magnetic Imager Instrument onboard the Solar Dynamics Observatory.

The Helioseismic and Magnetic Imager (HMI) instrument is a major component of NASA's Solar Dynamics Observatory (SDO) spacecraft. Since commencement of full regular science operations on 1 May 2010, HMI has operated with remarkable continuity, e.g. during the more than five years of the SDO prime mission that ended 30 September 2015, HMI collected 98.4% of all possible 45-second velocity maps; minimizing gaps in these full-disk Dopplergrams is crucial for helioseismology. HMI velocity, intensity, and magnetic-field measurements are used in numerous investigations, so understanding the quality of the data is important. This article describes the calibration measurements used to track the performance of the HMI instrument, and it details trends in important instrument parameters during the prime mission. Regular calibration sequences provide information used to improve and update the calibration of HMI data. The set-point temperature of the instrument front window and optical bench is adjusted regularly to maintain instrument focus, and changes in the temperature-control scheme have been made to improve stability in the observable quantities. The exposure time has been changed to compensate for a 20% decrease in instrument throughput. Measurements of the performance of the shutter and tuning mechanisms show that they are aging as expected and continue to perform according to specification. Parameters of the tunable optical-filter elements are regularly adjusted to account for drifts in the central wavelength. Frequent measurements of changing CCD-camera characteristics, such as gain and flat field, are used to calibrate the observations. Infrequent expected events such as eclipses, transits, and spacecraft off-points interrupt regular instrument operations and provide the opportunity to perform additional calibration. Onboard instrument anomalies are rare and seem to occur quite uniformly in time. The instrument continues to perform very well.

Circulating cell-free DNA as predictor of treatment failure after neoadjuvant chemo-radiotherapy before surgery in patients with locally advanced rectal cancer.

Background: Treatment of patients with locally advanced rectal cancer (LARC) is based on a combination of chemo-radiotherapy (CRT) and surgery. The rate of distant recurrences remains over 25%. Circulating cell-free DNA (cfDNA) in plasma is a mixture of normal and cancer-specific DNA segments and is a promising biomarker in patients with colorectal cancer. The aim of our study was to investigate plasma cfDNA as a prognostic marker for outcome in patients with LARC treated with neoadjuvant CRT and surgery. Patients and methods: In total, 123 patients with LARC were included in 2 biomarker studies. Patients were treated with neoadjuvant CRT before TME surgery. Fifty-two (42%) of the patients received induction chemotherapy with capecitabine + oxaliplatin. Total cfDNA was measured by direct fluorescent assay in EDTA plasma samples obtained at baseline, after induction chemotherapy, and after CRT. Serial samples 5 years after surgery were collected in 51 patients (41%). Results: Median follow-up was 55 months. Distant or local recurrence was seen in 30.9% of the patients. Patients with baseline cfDNA levels above the 75th quartile had a higher risk of local or distant recurrence and shorter time to recurrence compared with patients with plasma cfDNA below the 75th percentile (HR = 2.48, 95% CI: 1.3-4.8, P = 0.007). The same applied to disease-free survival (DFS) (HR = 2.43, 95% CI: 1.27-4.7, P = 0.015). In multivariate analysis, a high cfDNA level was significantly associated with time to progression and DFS. During follow-up, the association remained significant regardless of time point for sample analysis. Conclusion: We have demonstrated an association between a high baseline plasma level of cfDNA and increased risk of recurrence, shorter time to recurrence, and shorter DFS in patients with LARC. Consequently, cfDNA could potentially improve pre- and post-treatment risk assessment and facilitate individualized therapy for patients with LARC.

Synthesis of ligand-free CZTS nanoparticles via a facile hot injection route.

Single-phase, ligand-free Cu2ZnSnS4 (CZTS) nanoparticles that can be dispersed in polar solvents are desirable for thin film solar cell fabrication, since water can be used as the solvent for the nanoparticle ink. In this work, ligand-free nanoparticles were synthesized using a simple hot injection method and the precursor concentration in the reaction medium was tuned to control the final product. The as-synthesized nanoparticles were characterized using various techniques, and were found to have a near-stoichiometric composition and a phase-pure kesterite crystal structure. No secondary phases were detected with Raman spectroscopy or scanning transmission electron microscopy energy dispersive x-ray spectroscopy. Furthermore, high resolution transmission electron microscopy showed large-sized nanoparticles with an average diameter of 23 nm ± 11 nm. This approach avoids all organic materials and toxic solvents that otherwise could hinder grain growth and limit the deposition techniques. In addition the synthesis route presented here results in nanoparticles of a large size compared to other ligand-free CZTS nanoparticles, due to the high boiling point of the solvents selected. Large particle size in CZTS nanoparticle solar cells may lead to a promising device performance. The results obtained demonstrate the suitability of the synthesized nanoparticles for application in low cost thin film solar cells.

Femtosecond-laser ablation dynamics of dielectrics: basics and applications for thin films.

Laser ablation of dielectrics by ultrashort laser pulses is reviewed. The basic interaction between ultrashort light pulses and the dielectric material is described, and different approaches to the modeling of the femtosecond ablation dynamics are reviewed. Material excitation by ultrashort laser pulses is induced by a combination of strong-field excitation (multi-photon and tunnel excitation), collisional excitation (potentially leading to an avalanche process), and absorption in the plasma consisting of the electrons excited to the conduction band. It is discussed how these excitation processes can be described by various rate-equation models in combination with different descriptions of the excited electrons. The optical properties of the highly excited dielectric undergo a rapid change during the laser pulse, which must be included in a detailed modeling of the excitations. The material ejected from the dielectric following the femtosecond-laser excitation can potentially be used for thin-film deposition. The deposition rate is typically much smaller than that for nanosecond lasers, but film production by femtosecond lasers does possess several attractive features. First, the strong-field excitation makes it possible to produce films of materials that are transparent to the laser light. Second, the highly localized excitation reduces the emission of larger material particulates. Third, lasers with ultrashort pulses are shown to be particularly useful tools for the production of nanocluster films. The important question of the film stoichiometry relative to that of the target will be thoroughly discussed in relation to the films reported in the literature.

Induction chemotherapy with capecitabine and oxaliplatin followed by chemoradiotherapy before total mesorectal excision in patients with locally advanced rectal cancer.

BACKGROUND: Preoperative chemoradiation in patients with locally advanced rectal cancer has no impact on overall survival (OS) and distant recurrences. The aim of the study was to evaluate local downstaging, toxicity and long-term outcome in patients with locally advanced rectal cancer after induction therapy with capecitabine and oxaliplatin (CAPEOX) followed by radiotherapy concomitant with capecitabine [chemoradiotherapy (CRT)] before total mesorectal excision (TME). PATIENTS AND METHODS: Patients with T4 tumors, all T3N+ tumors or T3 tumors involving or with a distance ≤1 mm to the mesorectal fascia were included. Patients were planned for two cycles of CAPEOX followed by radiotherapy concomitant with capecitabine. TME was carried out 6 weeks after the completion of CRT. RESULTS: Of 84 consecutively admitted patients starting induction CAPEOX, 77 patients underwent surgery. R0 resection was seen in 94% and T downstaging in 69%. In the intention-to-treat group, pathological complete response was seen in 23%. Five-year disease-free survival (DFS) and OS were 63% [95% confidence interval (CI), 52.2% to 73.7%] and 67% (95% CI, 56.1% to 77.3%), respectively. Grade 3/4 toxicity was seen in 18%, and four deaths occurred within 2 months of therapy. CONCLUSION: Induction chemotherapy before CRT and surgery showed a high local control rate and promising long-term outcome as OS and DFS.

The T allele of rs7903146 TCF7L2 is associated with impaired insulinotropic action of incretin hormones, reduced 24 h profiles of plasma insulin and glucagon, and increased hepatic glucose production in young healthy men.

AIMS/HYPOTHESIS: We studied the physiological, metabolic and hormonal mechanisms underlying the elevated risk of type 2 diabetes in carriers of TCF7L2 gene. METHODS: We undertook genotyping of 81 healthy young Danish men for rs7903146 of TCF7L2 and carried out various beta cell tests including: 24 h glucose, insulin and glucagon profiles; OGTT; mixed meal test; IVGTT; hyperglycaemic clamp with co-infusion of glucagon-like peptide (GLP)-1 or glucose-dependent insulinotropic polypeptide (GIP); and a euglycaemic-hyperinsulinaemic clamp combined with glucose tracer infusion to study hepatic and peripheral insulin action. RESULTS: Carriers of the T allele were characterised by reduced 24 h insulin concentrations (p < 0.05) and reduced insulin secretion relative to glucose during a mixed meal test (beta index: p < 0.003), but not during an IVGTT. This was further supported by reduced late-phase insulinotropic action of GLP-1 (p = 0.03) and GIP (p = 0.07) during a 7 mmol/l hyperglycaemic clamp. Secretion of GLP-1 and GIP during the mixed meal test was normal. Despite elevated hepatic glucose production, carriers of the T allele had significantly reduced 24 h glucagon concentrations (p < 0.02) suggesting altered alpha cell function. CONCLUSIONS/INTERPRETATION: Elevated hepatic glucose production and reduced insulinotropic effect of incretin hormones contribute to an increased risk of type 2 diabetes in carriers of the rs7903146 risk T allele of TCF7L2.

Wave-like properties of solar supergranulation.

Supergranulation on the surface of the Sun is a pattern of horizontal outflows, outlined by a network of small magnetic features, with a distinct scale of 30 million metres and an apparent lifetime of one day. It is generally believed that supergranulation corresponds to a preferred 'cellular' scale of thermal convection; rising magnetic fields are dragged by the outflows and concentrated into 'ropes' at the 'cell' boundaries. But as the convection zone is highly turbulent and stratified, numerical modelling has proved to be difficult and the dynamics remain poorly understood. Moreover, there is as yet no explanation for the observation that the pattern appears to rotate faster around the Sun than the magnetic features. Here we report observations showing that supergranulation undergoes oscillations and supports waves with periods of 6-9 days. The waves are predominantly prograde, which explains the apparent super-rotation of the pattern. The rotation of the plasma through which the pattern propagates is consistent with the motion of the magnetic network.

Helioseismic measurement of solar torsional oscillations.

Bands of slower and faster rotation, the so-called torsional oscillations, are observed at the Sun's surface to migrate in latitude over the 11-year solar cycle. Here, we report on the temporal variations of the Sun's internal rotation from solar p-mode frequencies obtained over nearly 6 years by the Michelson Doppler Imager (MDI) instrument on board the Solar and Heliospheric Observatory (SOHO) satellite. The entire solar convective envelope appears to be involved in the torsional oscillations, with phase propagating poleward and equatorward from midlatitudes at all depths throughout the convective envelope.

In search of the most suitable technique for truncal spinal immobilization with associated radiography.

The spineboard (SB) and the vacuum mattress (VM) are utilized for prehospital and emergency department (ED) immobilization of the spine. While permitting excellent pictures to be taken, the SB is a very painful device that can only be used for a limited time. The current study investigated the feasibility of different models of the VM for radiography. Computed tomography for each of seven VMs showed different degrees of shadows from the sac containing the polystyrole balls. This was related first of all to a high contrast of the sac in comparison with its contents and to a sometimes considerable shrinkage of the latter that resulted in further folding into the sac and, secondly, also to very broad mattresses, additional chambers within the VM and various grips and supports for lifting the mattress. Therefore, some features designed for prehospital use of the VM are actually acting against its use for diagnostic purposes. The aim of this study was to discuss and identify possible properties of a device that is useful for both immobilization and diagnostic purposes.

Angular distribution of electron temperature and density in a laser-ablation plume

The angular distribution of electron temperature and density in a laser-ablation plume has been studied for the first time. The electron temperature ranges from 0.1 to 0.5 eV and is only weakly dependent on the angle in the low-intensity range studied here. In contrast, the typical ion energy is about 2 orders of magnitude larger, and its angular distribution is more peaked about the target normal. The derived values of the electron density are in agreement with the measured values of ion density.

Changes in sexual behavior after orthopedic replacement of hip or knee in elderly males--a prospective study.

The aim of this study was to investigate two questions: Does arthroplastic surgery affect the patient's status as being sexually active; and if patients are sexually active, does surgery affect their erectile function? The study was designed prospectively and the patients filled in a questionnaire concerning sexuality and erectile function before and 6 months after alloplastic hip-or knee surgery. Ninety-nine males were included, mean age 70.6 y. The results demonstrate that 17% of patients lost a sexual activity that they had preoperatively, and no one regained sexual activity after surgery. A correlation between increasing age and risk of losing sexual activity was demonstrated. 26.1% lost a normal erectile function they had preoperatively, while 6.7% regained normal erections. A similar correlation between increasing age and increased risk was demonstrated. From this study of elderly males undergoing orthopaedic alloplastic surgery it is concluded that the risk of losing sexual activity and erectile capability is increased after surgery, and especially in the group where sexual functions are already impaired.

Deeply Penetrating Banded Zonal Flows in the Solar Convection Zone.

Helioseismic observations have detected small temporal variations of the rotation rate below the solar surface that correspond to the so-called "torsional oscillations" known from Doppler measurements of the surface. These appear as bands of slower- and faster-than-average rotation moving equatorward. Here we establish, using complementary helioseismic observations over 4 yr from the GONG network and from the MDI instrument on board SOHO, that the banded flows are not merely a near-surface phenomenon: rather, they extend downward at least 60 Mm (some 8% of the total solar radius) and thus are evident over a significant fraction of the nearly 200 Mm depth of the solar convection zone.

Dynamic variations at the base of the solar convection zone

We have detected changes in the rotation of the sun near the base of its convective envelope, including a prominent variation with a period of 1.3 years at low latitudes. Such helioseismic probing of the deep solar interior has been enabled by nearly continuous observation of its oscillation modes with two complementary experiments. Inversion of the global-mode frequency splittings reveals that the largest temporal changes in the angular velocity Omega are of the order of 6 nanohertz and occur above and below the tachocline that separates the sun's differentially rotating convection zone (outer 30% by radius) from the nearly uniformly rotating deeper radiative interior beneath. Such changes are most pronounced near the equator and at high latitudes and are a substantial fraction of the average 30-nanohertz difference in Omega with radius across the tachocline at the equator. The results indicate variations of rotation close to the presumed site of the solar dynamo, which may generate the 22-year cycles of magnetic activity.