Ultra-low oxygen, liquid sample cell for in situ synchrotron-based small-wide angle scattering (SAXS-WAXS).

Here, we report the design and successful implementation of an ultra-low oxygen sample cell for use on the SAXS-WAXS (small-wide angle x-ray scattering) beamline I22 at DIAMOND. The rigorous exclusion of oxygen is found to require double jacketing with purge gas throughout the entire system, pipework, pumps, and the sample cell itself. This particularly includes a "double-window" arrangement at the sample location to accommodate the very tight geometrical restrictions of the sample position. The in situ cell design also requires the additional complexity of heating the sample/solution and real-time electrochemical measurements. We demonstrate the successful implementation of this arrangement with real-time in situ characterization of an iron foil corrosion evolving under the "sweet-scale environment," very anoxic conditions common, in particular, commercial situations. The formation of iron carbonate, siderite, rather than iron oxide, indicates that our system is oxygen free down very low levels (<35 ppb at 80 °C).

Prognostic value of plasma circulating tumor DNA fraction across four common cancer types: a real-world outcomes study.

BACKGROUND: Genomic analysis of circulating tumor DNA (ctDNA) is increasingly incorporated into the clinical management of patients with advanced cancer. Beyond tumor profiling, ctDNA analysis also can enable calculation of circulating tumor fraction (TF), which has previously been found to be prognostic. While most prognostic models in metastatic cancer are tumor type specific and require significant patient-level data, quantification of TF in ctDNA has the potential to serve as a pragmatic, tumor-agnostic prognostic tool. PATIENTS AND METHODS: This study utilized a cohort of patients in a nationwide de-identified clinico-genomic database with metastatic castration-resistant prostate cancer (mCRPC), metastatic breast cancer (mBC), advanced non-small-cell lung cancer (aNSCLC), or metastatic colorectal cancer (mCRC) undergoing liquid biopsy testing as part of routine care. TF was calculated based on single-nucleotide polymorphism aneuploidy across the genome. Clinical, disease, laboratory, and treatment data were captured from the electronic health record. Overall survival (OS) was evaluated by TF level while controlling for relevant covariables. RESULTS: A total of 1725 patients were included: 198 mCRPC, 402 mBC, 902 aNSCLC, and 223 mCRC. TF ≥10% was highly correlated with OS in univariable analyses for all cancer types: mCRPC [hazard ratio (HR) 3.3, 95% confidence interval (CI) 2.04-5.34, P < 0.001], mBC (HR 2.4, 95% CI 1.71-3.37, P < 0.001), aNSCLC (HR 1.68, 95% CI 1.34-2.1, P < 0.001), and mCRC (HR 2.11, 95% CI 1.39-3.2, P < 0.001). Multivariable assessments of TF had similar point estimates and CIs, suggesting a consistent and independent association with survival. Exploratory analysis showed that TF remained consistently prognostic across a wide range of cutpoints. CONCLUSIONS: Plasma ctDNA TF is a pragmatic, independent prognostic biomarker across four advanced cancers with potential to guide clinical conversations around expected treatment outcomes. With further prospective validation, ctDNA TF could be incorporated into care paradigms to enable precision escalation and de-escalation of cancer therapy based on patient-level tumor biology.

I22: SAXS/WAXS beamline at Diamond Light Source - an overview of 10 years operation.

Beamline I22 at Diamond Light Source is dedicated to the study of soft-matter systems from both biological and materials science. The beamline can operate in the range 3.7 keV to 22 keV for transmission SAXS and 14 keV to 20 keV for microfocus SAXS with beam sizes of 240 µm × 60 µm [full width half-maximum (FWHM) horizontal (H) × vertical (V)] at the sample for the main beamline, and approximately 10 µm × 10 µm for the dedicated microfocusing platform. There is a versatile sample platform for accommodating a range of facilities and user-developed sample environments. The high brilliance of the insertion device source on I22 allows structural investigation of materials under extreme environments (for example, fluid flow at high pressures and temperatures). I22 provides reliable access to millisecond data acquisition timescales, essential to understanding kinetic processes such as protein folding or structural evolution in polymers and colloids.

The modular small-angle X-ray scattering data correction sequence.

Data correction is probably the least favourite activity amongst users experimenting with small-angle X-ray scattering: if it is not done sufficiently well, this may become evident only during the data analysis stage, necessitating the repetition of the data corrections from scratch. A recommended comprehensive sequence of elementary data correction steps is presented here to alleviate the difficulties associated with data correction, both in the laboratory and at the synchrotron. When applied in the proposed order to the raw signals, the resulting absolute scattering cross section will provide a high degree of accuracy for a very wide range of samples, with its values accompanied by uncertainty estimates. The method can be applied without modification to any pinhole-collimated instruments with photon-counting direct-detection area detectors.

Cytomegalic interneurons: a new abnormal cell type in severe pediatric cortical dysplasia.

A defining histopathologic feature of Taylor-type cortical dysplasia (CD) is the presence of cytomegalic neurons and balloon cells. Most cytomegalic neurons appear to be pyramidal-shaped and glutamatergic. The present study demonstrates the presence of cytomegalic GABAergic interneurons in a subset of pediatric patients with severe CD. Cortical tissue samples from children with mild, severe, and non-CD pathologies were examined using morphologic and electrophysiologic techniques. By using in vitro slices, cytomegalic cells with characteristics consistent with interneurons were found in 6 of 10 patients with severe CD. Biocytin labeling demonstrated that cytomegalic interneurons had more dendrites than normal-appearing interneurons. Whole-cell patch clamp recordings showed that cytomegalic interneurons had increased membrane capacitance and time constant compared with normal-appearing interneurons. They also displayed signs of cellular hyperexcitability, evidenced by increased firing rates, decreased action potential inactivation, and the occurrence of spontaneous membrane depolarizations. Single-cell reverse transcription-polymerase chain reaction and immunohistochemistry for GABAergic markers provided further evidence that these cells were probably cytomegalic interneurons. The pathophysiologic significance of GABAergic cytomegalic interneurons in severe CD tissue is unknown, but they could inhibit glutamatergic cytomegalic pyramidal neurons, or contribute to the synchronization of neuronal networks and the propagation of ictal activity in a subset of pediatric patients with severe CD.

Improving MRI differentiation of gray and white matter in epileptogenic lesions based on nonlinear feedback.

A new method for enhancing MRI contrast between gray matter (GM) and white matter (WM) in epilepsy surgery patients with symptomatic lesions is presented. This method uses the radiation damping feedback interaction in high-field MRI to amplify contrast due to small differences in resonance frequency in GM and WM corresponding to variations in tissue susceptibility. High-resolution radiation damping-enhanced (RD) images of in vitro brain tissue from five patients were acquired at 14 T and compared with corresponding conventional T(1)-, T(2) (*)-, and proton density (PD)-weighted images. The RD images yielded a six times better contrast-to-noise ratio (CNR = 44.8) on average than the best optimized T(1)-weighted (CNR = 7.92), T(2) (*)-weighted (CNR = 4.20), and PD-weighted images (CNR = 2.52). Regional analysis of the signal as a function of evolution time and initial pulse flip angle, and comparison with numerical simulations confirmed that radiation damping was responsible for the observed signal growth. The time evolution of the signal in different tissue regions was also used to identify subtle changes in tissue composition that were not revealed in conventional MR images. RD contrast is compared with conventional MR methods for separating different tissue types, and its value and limitations are discussed.

Contralateral hemimicrencephaly and clinical-pathological correlations in children with hemimegalencephaly.

In paediatric epilepsy surgery patients with hemimegalencephaly (HME; n = 23), this study compared clinical, neuroimaging and pathologic features to discern potential mechanisms for suboptimal post-hemispherectomy developmental outcomes and structural pathogenesis. MRI measured affected and non-affected cerebral hemisphere volumes for HME and non-HME cases, including monozygotic twins where one sibling had HME. Staining against neuronal nuclei (NeuN) determined grey and white matter cell densities and sizes in HME and autopsy cases, including the non-affected side of a HME surgical/autopsy case. By MRI, the affected hemisphere was larger and the non-affected side smaller in HME compared with non-HME children. The affected HME side showed enlarged abnormal deep grey and white matter structures and/or T2-weighted hypointensity in the subcortical white matter in 75% of cases, suggestive of excessive pre-natal neurogenesis and heterotopias. Histopathological examination of the affected HME side revealed immature-appearing neurons in 70%, polymicrogyria (PMG) in 61% and balloon cells in 45% of cases. Compared with autopsy cases, in HME children NeuN cell densities on the affected side were increased in the molecular layer and upper cortex (+244 to +18%), decreased in lower cortical layers (-35%) and increased in the white matter (+139 to +149%). Deep grey matter MRI abnormalities and/or T2-weighted white matter hypointensity correlated with the presence of immature-appearing neurons and PMG on histopathology, decreased NeuN cell densities in lower cortical layers and a positive history of infantile spasms. Post-surgery seizure control was associated with decreased NeuN densities in the molecular layer. In young children with HME and epilepsy, these findings indicate that there are bilateral cerebral hemispheric abnormalities and contralateral hemimicrencephaly is a likely explanation for poorer post-surgery seizure control and cognitive outcomes. In addition, our findings support the hypothesis that HME pathogenesis probably involves somatic mutations that affect each developing cerebral hemisphere differently with more neurons than expected on the HME side.

Cerebral hemispherectomy in pediatric patients with epilepsy: comparison of three techniques by pathological substrate in 115 patients.

OBJECT: Cerebral hemispherectomy for intractable seizures has evolved over the past 50 years, and current operations focus less on brain resection and more on disconnection. In addition, cases involving cortical dysplasia and Rasmussen encephalitis are being identified and surgically treated in younger individuals. Few studies have been conducted to compare whether there are perioperative differences based on hemispherectomy technique and/or pathological substrate in pediatric patients with epilepsy. METHODS: In this study the authors compared, stratified by disease, anatomical (37 cases) and Rasmussen functional hemispherectomy (32 cases) with a new modified lateral hemispherotomy (46 cases). Pathological processes included cortical dysplasia (55 cases), Rasmussen encephalitis (21 cases), infarction/ischemia (27 cases), and other/miscellaneous (12 cases). The authors found differences in perioperative clinical factors based on operative technique and/or pathological substrate. In terms of technique, the lateral hemispherotomy was associated with the least intraoperative blood loss, shortest intensive care unit stay, and lowest complication rate. The anatomical hemispherectomy was associated with the longest hospital stay, delayed oral food intake, highest postsurgery fevers, and the highest incidence of shunt requirement. The functional hemispherectomy was associated with the highest reoperation rate for recurrent seizures (25%). In terms of pathology, patients with cortical dysplasia were the youngest at surgery, suffered the greatest amount of blood loss, and required the longest operative/anesthesia times compared with the other pathologically defined groups. Postoperative seizure control (range 0.5-2 years) was not statistically different according to technique or disease process and was similar to that in cases of pediatric temporal lobe epilepsy. CONCLUSIONS: The authors found differences in perioperative risks and hospital course but not postsurgery seizure control, which vary by hemispherectomy technique and/or disease process. The modified lateral hemispherotomy approach offers various advantages related to operative blood loss and reoperation compared with anatomical and functional hemispherectomies that are especially relevant in younger patients with cortical dysplasia and Rasmussen encephalitis with small and/or malformed ventricles.

Enhanced expression of a specific hyperpolarization-activated cyclic nucleotide-gated cation channel (HCN) in surviving dentate gyrus granule cells of human and experimental epileptic hippocampus.

Changes in the expression of ion channels, contributing to altered neuronal excitability, are emerging as possible mechanisms in the development of certain human epilepsies. In previous immature rodent studies of experimental prolonged febrile seizures, isoform-specific changes in the expression of hyperpolarization-activated cyclic nucleotide-gated cation channels (HCNs) correlated with long-lasting hippocampal hyperexcitability and enhanced seizure susceptibility. Prolonged early-life seizures commonly precede human temporal lobe epilepsy (TLE), suggesting that transcriptional dysregulation of HCNs might contribute to the epileptogenic process. Therefore, we determined whether HCN isoform expression was modified in hippocampi of individuals with TLE. HCN1 and HCN2 expression were measured using in situ hybridization and immunocytochemistry in hippocampi from three groups: TLE with hippocampal sclerosis (HS; n = 17), epileptic hippocampi without HS, or non-HS (NHS; n = 10), and autopsy material (n = 10). The results obtained in chronic human epilepsy were validated by examining hippocampi from the pilocarpine model of chronic TLE. In autopsy and most NHS hippocampi, HCN1 mRNA expression was substantial in pyramidal cell layers and lower in dentate gyrus granule cells (GCs). In contrast, HCN1 mRNA expression over the GC layer and in individual GCs from epileptic hippocampus was markedly increased once GC neuronal density was reduced by >50%. HCN1 mRNA changes were accompanied by enhanced immunoreactivity in the GC dendritic fields and more modest changes in HCN2 mRNA expression. Furthermore, similar robust and isoform-selective augmentation of HCN1 mRNA expression was evident also in the pilocarpine animal model of TLE. These findings indicate that the expression of HCN isoforms is dynamically regulated in human as well as in experimental hippocampal epilepsy. After experimental febrile seizures (i.e., early in the epileptogenic process), the preserved and augmented inhibition onto principal cells may lead to reduced HCN1 expression. In contrast, in chronic epileptic HS hippocampus studied here, the profound loss of interneuronal and principal cell populations and consequent reduced inhibition, coupled with increased dendritic excitation of surviving GCs, might provoke a "compensatory" enhancement of HCN1 mRNA and protein expression.

Effect of race and resistance training status on the density of fat-free mass and percent fat estimates.

The impact of race and resistance training status on the assumed density of the fat-free mass (D(FFM)) and estimates of body fatness via hydrodensitometry (%Fat(D)) vs. a four-component model (density, water, mineral; %Fat(D,W,M)) were determined in 45 men: white controls (W; n = 15), black controls (B; n = 15), and resistance-trained blacks (B-RT; n = 15). Body density by hydrostatic weighing, body water by deuterium dilution, and bone mineral by dual-energy X-ray absorptiometry were used to estimate %Fat(D,W,M). D(FFM) was not different between B and W (or 1.1 g/ml); however, D(FFM) in B-RT was significantly lower (1.091 +/- 0.012 g/ml; P < 0.05). Therefore, %Fat(D) using the Siri equation was not different from %Fat(D,W,M) in W (17.5 +/- 5.0 vs. 18.3 +/- 5.4%) or B (14.9 +/- 5.6 vs. 15.7 +/- 5.7%) but significantly overestimated %Fat(D,W,M) in B-RT (14.0 +/- 5.9 vs. 10.4 +/- 6.0%; P < 0.05). The use of a race-specific equation (assuming D(FFM) = 1.113 g/ml) did not improve the agreement between %Fat(D) and %Fat(D,W,M), resulting in a significantly greater mean (+/-SD) discrepancy for B (1.7 +/- 1.8% fat) and B-RT (6.2 +/- 4.3% fat). Thus race per se does not affect D(FFM) or estimates of %Fat(D); however, B-RT have a D(FFM) lower than 1.1 g/ml, leading to an overestimation of %Fat(D).

Use of air displacement plethysmography for estimating body fat in a four-component model.

PURPOSE: To compare measurements of body density (D(b)) obtained from air displacement plethysmography (AP) and hydrostatic weighing (HW) and to determine the accuracy of substituting D(b) via AP (D(b)-AP) for D(b) via HW (D(b)-HW) in estimating body fatness (%Fat(4C)) and the composition and density of the fat-free mass (Dffm) from a four-component model (fat, mineral, water, and protein). METHODS: D(b) was measured in 50 young adults using AP and HW. Total body water via deuterium dilution, bone mineral content via dual-energy x-ray absorptiometry, and D(b) were used to estimate %Fat(4C). RESULTS: D(b)-AP and D(b)-HW were highly correlated (r = 0.89, SEE = 0.008 g x mL(-1)), but D(b)-AP (1.065 +/- 0.003 g x mL(-1)) was significantly higher (P < 0.05) than D(b)-HW (1.058 +/- 0.003 g x mL(-1)), resulting in a mean difference of 2.8%fat. Differences between %Fat(4C-AP) (17.8 +/- 1.2%) and %Fat(4C-HW) (19.3 +/- 1.2%) were significant (P < 0.05), but the SD of the differences (2.3%) was low. When D(b)-AP was used in a four-component model in place of D(b)-HW, the calculated Dffm was significantly higher (1.109 +/- 0.002 vs 1.105 +/- 0.002 g x mL(-1)) based on a higher (P < 0.05) protein fraction (22.0 +/- 0.4% vs 20.6 +/- 0.4%) and lower (P < 0.05) water (71.1 +/- 0.4% vs 72.4 +/- 0.4%) and mineral fractions (7.0 +/- 0.1% vs 7.1 +/- 0.1%). CONCLUSIONS: AP yields a higher D(b) than HW and may not be a valid method for measuring D(b) or estimating %fat using densitometry. However, due to relatively small bias and low individual error, D(b)-AP is an acceptable substitute for D(b)-HW when estimating %fat with a four-component model in young adults.

The metabolism of beta-chloroprene: preliminary in-vitro studies using liver microsomes.

Based on analogy with butadiene and isoprene, the metabolism of beta-chloroprene (2-chloro-1,3-butadiene, CD) to reactive intermediates is likely to be a key determinant of tumor development in laboratory rodents exposed to CD by inhalation. The purpose of this study is to identify species differences in toxic metabolite (epoxide) formation and detoxification in rodents and humans. The in-vitro metabolism of CD was studied in liver microsomes of B6C3F1 mice, Fischer/344 and Wistar rats, Syrian hamsters, and humans. Microsomal oxidation of CD in the presence of NADP(+), extraction with diethyl ether, and analysis by GC-mass selective detection (MSD) indicated that (1-chloroethenyl)oxirane (CEO) was an important metabolite of CD in the liver microsomal suspensions of all species studied. Other potential water-soluble oxidative metabolites may have been present. The oxidation of CD was inhibited by 4-methyl pyrazole, an inhibitor of CYP 2E1. CEO was sufficiently volatile at 37 degrees C for vial headspace analysis using GC-MSD single ion monitoring (m/z=39). CEO was synthesized and used to conduct partition measurements along with CD and further explore CEO metabolism in liver microsomes and cytosol. The liquid-to-air partition coefficients for CD and CEO in the microsomal suspensions were 0.7 and 58, respectively. Apparent species differences in the uptake of CEO by microsomal hydrolysis were hamster approximately human>rats>mice. Hydrolysis was inhibited by 1,1,1-trichloropropene oxide, a competitive inhibitor of epoxide hydrolase. A preliminary experiment indicated that the uptake of CEO in liver cytosol by GSH conjugation was hamster>rats approximately mice (human cytosol not yet tested). In general, the results suggest that metabolism may help explain species differences showing a greater sensitivity for CD-induced tumorigenicity in mice, for example, compared with hamsters. Additional experiments are in progress to quantify the kinetic parameters of CD oxidation and CEO metabolism by enzymatic hydrolysis and conjugation by glutathione S-transferase for in cytosol. A future goal is to use the kinetic rates to parameterize a physiologically based toxicokinetic model and relate the burden of toxic metabolite to the cancer dose-response observed in experimental animals.

The unidentified diffuse interstellar bands as evidence for large organic molecules in the interstellar medium.

Since 1921, astronomers have been aware of a set of spectral absorption features that are formed in interstellar space but are not identifiable with any known atomic or molecular species. Today some 200 of these so-called diffuse interstellar bands (DIBs) are known, yet none has yet been identified unambiguously with any specific carrier. Recent evidence from astronomical observations, laboratory spectroscopy, and chemical studies suggests that the responsible species are large carbon-bearing molecules. This paper provides a brief history of the DIB problem, a review of suggested carriers, and a description and discussion of new results from the telescope and from laboratory measurements of chemical reaction rates in candidate carriers.

Alternative uses for the haemostatic vascular sheath.

The haemostatic vascular sheath has several non-vascular applications in interventional radiology. It is used to preserve access to an organ (i.e. contrast can be introduced without removing catheters and guidewires). There are also advantages regarding patient comfort and safety. Three such applications are described.

Serum cholesterol levels in college students: opportunities for education and intervention.

Elevated serum cholesterol levels have been shown to be associated with premature atherosclerosis in adolescents and young adults. The National Cholesterol Education Program recommends cholesterol screening for all adults aged 20 years or older, but normative data on the college-age population are limited. At a university where lipid profiles are made available to students in selected health/wellness courses, the authors analyzed and summarized lipid profiles on 1,088 undergraduates. Mean total cholesterol levels were similar for men (165 +/- 33 mg/dL) and women (168 +/- 27 mg/dL). The men, however, had significantly lower high-density lipoprotein (HDL) cholesterol and higher low-density lipoprotein (LDL) cholesterol than the women. One hundred twenty-one students (11.1% of the sample) had elevated serum cholesterol levels (LDL-C > or = 130 mg/dL). Cholesterol screening can be used as an educational tool for college students to reinforce the link between lipid levels and health habits.

An infrared spectral match between GEMS and interstellar grains.

Infrared spectral properties of silicate grains in interplanetary dust particles (IDPs) were compared with those of astronomical silicates. The approximately 10-micrometer silicon-oxygen stretch bands of IDPs containing enstatite (MgSiO3), forsterite (Mg2SiO4), and glass with embedded metal and sulfides (GEMS) exhibit fine structure and bandwidths similar to those of solar system comets and some pre-main sequence Herbig Ae/Be stars. Some GEMS exhibit a broad, featureless silicon-oxygen stretch band similar to those observed in interstellar molecular clouds and young stellar objects. These GEMS provide a spectral match to astronomical "amorphous" silicates, one of the fundamental building blocks from which the solar system is presumed to have formed.

Evaluation of the BOD POD for assessing body fat in collegiate football players.

PURPOSE: The purpose of this investigation was to evaluate the accuracy of a new air displacement plethysmograph, BOD POD Body Composition System, for determining %fat in collegiate football players. METHODS: Body fatness was estimated from body density (Db), which was measured on the same day using the BOD POD and hydrostatic weighing (HW) in 69 Division IA football players. In addition, 20 subjects were whole body scanned using dual-energy x-ray absorptiometry, DXA (Lunar DPX-L) to assess total body mineral content and %fat. Mineral content and HW determined Db were used to compute %fat from a three-component model (3C; fat, mineral, and residual). RESULTS: Test-retest reliability for assessing %fat using the BOD POD (N = 15) was 0.994 with a technical error of measurement of 0.448%. Mean (+/- SEM) Db measured with the BOD POD (1.064 +/- 0.002 g x cc(-1) was significantly greater (P < 0.05) than HW (1.060 +/- 0.002 g x cc(-1)), thus resulting in a lower %fat for the BOD POD (15.1 +/- 0.8%) compared with HW (17.0 +/- 0.8%). Similar results (N = 20) were found for DXA (12.9 +/- 1.2%) and the 3C (12.7 +/- 0.8%) where %fat scores were significantly higher (P < 0.05) than scores determined using the BOD POD (10.9 +/- 1.0%). CONCLUSIONS: Db measured with the BOD POD was higher than the criterion HW, thus yielding lower %fat scores for the BOD POD. In addition, BOD POD determined %fat was lower than DXA and 3C determined values in a subgroup of subjects. Assessment of %fat using the BOD POD is reliable and requires minimal technical expertise; however, in this study of collegiate football players, %fat values were underpredicted when compared to HW, DXA, and the 3C model.

Transcolonic retrograde ureteric catheterization assisted by 3-D computed tomography.

A 42-year-old woman had bladder exstrophy at birth, treated by ureterocolic anastomosis of her single kidney. She suffered recurrent hyperammonaemia, leading to comas, but refused an ileal conduit. During her most recent coma, it was decided to divert her urine to test whether this would reduce hyperammonaemia: this was accomplished by transcolonic retrograde catheterization of the ureter. This was only possible after computed tomography ureterography to show the ureterocolic anastomosis.