A new Kirkpatrick-Baez-based scanning microscope for the Submicron Resolution X-ray Spectroscopy (SRX) beamline at NSLS-II.

The development, construction, and first commissioning results of a new scanning microscope installed at the 5-ID Submicron Resolution X-ray Spectroscopy (SRX) beamline at NSLS-II are reported. The developed system utilizes Kirkpatrick-Baez mirrors for X-ray focusing. The instrument is designed to enable spectromicroscopy measurements in 2D and 3D with sub-200 nm spatial resolution. The present paper focuses on the design aspects, optical considerations, and specifics of the sample scanning stage, summarizing some of the initial commissioning results.

Augmented decision-making for acral lentiginous melanoma detection using deep convolutional neural networks.

BACKGROUND: Several studies have achieved high-level performance of melanoma detection using convolutional neural networks (CNNs). However, few have described the extent to which the implementation of CNNs improves the diagnostic performance of the physicians. OBJECTIVE: This study is aimed at developing a CNN for detecting acral lentiginous melanoma (ALM) and investigating whether its implementation can improve the initial decision for ALM detection made by the physicians. METHODS: A CNN was trained using 1072 dermoscopic images of acral benign nevi, ALM and intermediate tumours. To investigate whether the implementation of CNN can improve the initial decision for ALM detection, 60 physicians completed a three-stage survey. In Stage I, they were asked for their decisions solely on the basis of dermoscopic images provided to them. In Stage II, they were also provided with clinical information. In Stage III, they were provided with the additional diagnosis and probability predicted by the CNN. RESULTS: The accuracy of ALM detection in the participants was 74.7% (95% confidence interval [CI], 72.6-76.8%) in Stage I and 79.0% (95% CI, 76.7-81.2%) in Stage II. In Stage III, it was 86.9% (95% CI, 85.3-88.4%), which exceeds the accuracy delivered in Stage I by 12.2%p (95% CI, 10.1-14.3%p) and Stage II by 7.9%p (95% CI, 6.0-9.9%p). Moreover, the concordance between the participants considerably increased (Fleiss-κ of 0.436 [95% CI, 0.437-0.573] in Stage I, 0.506 [95% CI, 0.621-0.749] in Stage II and 0.684 [95% CI, 0.621-0.749] in Stage III). CONCLUSIONS: Augmented decision-making improved the performance of and concordance between the clinical decisions of a diverse group of experts. This study demonstrates the potential use of CNNs as an adjoining, decision-supporting system for physicians' decisions.

Multimodal X-ray imaging of grain-level properties and performance in a polycrystalline solar cell.

The factors limiting the performance of alternative polycrystalline solar cells as compared with their single-crystal counterparts are not fully understood, but are thought to originate from structural and chemical heterogeneities at various length scales. Here, it is demonstrated that multimodal focused nanobeam X-ray microscopy can be used to reveal multiple aspects of the problem in a single measurement by mapping chemical makeup, lattice structure and charge collection efficiency simultaneously in a working solar cell. This approach was applied to micrometre-sized individual grains in a Cu(In,Ga)Se2 polycrystalline film packaged in a working device. It was found that, near grain boundaries, collection efficiency is increased, and that in these regions the lattice parameter of the material is expanded. These observations are discussed in terms of possible physical models and future experiments.

Design and performance of an X-ray scanning microscope at the Hard X-ray Nanoprobe beamline of NSLS-II.

A hard X-ray scanning microscope installed at the Hard X-ray Nanoprobe beamline of the National Synchrotron Light Source II has been designed, constructed and commissioned. The microscope relies on a compact, high stiffness, low heat dissipation approach and utilizes two types of nanofocusing optics. It is capable of imaging with ∼15 nm × 15 nm spatial resolution using multilayer Laue lenses and 25 nm × 26 nm resolution using zone plates. Fluorescence, diffraction, absorption, differential phase contrast, ptychography and tomography are available as experimental techniques. The microscope is also equipped with a temperature regulation system which allows the temperature of a sample to be varied in the range between 90 K and 1000 K. The constructed instrument is open for general users and offers its capabilities to the material science, battery research and bioscience communities.

Pushing the limits: an instrument for hard X-ray imaging below 20 nm.

Hard X-ray microscopy is a prominent tool suitable for nanoscale-resolution non-destructive imaging of various materials used in different areas of science and technology. With an ongoing effort to push the 2D/3D imaging resolution down to 10 nm in the hard X-ray regime, both the fabrication of nano-focusing optics and the stability of the microscope using those optics become extremely challenging. In this work a microscopy system designed and constructed to accommodate multilayer Laue lenses as nanofocusing optics is presented. The developed apparatus has been thoroughly characterized in terms of resolution and stability followed by imaging experiments at a synchrotron facility. Drift rates of ∼2 nm h(-1) accompanied by 13 nm × 33 nm imaging resolution at 11.8 keV are reported.

X-ray microscopy and tomography detect the accumulation of bare and PEG-coated gold nanoparticles in normal and tumor mouse tissues.

We demonstrate that, with appropriate staining, high-resolution X-ray microscopy can image complicated tissue structures--cerebellum and liver--and resolve large or small amounts of Au nanoparticles in these tissues. Specifically, images of tumor tissue reveal high concentrations of accumulated Au nanoparticles. PEG (poly(ethylene glycol)) coating is quite effective in enhancing this accumulation and significantly modifies the mechanism of uptake by reticuloendothelial system (RES) organs.

The modified Devine's procedure for the management of concealed penis in children: an experience of 131 cases.

OBJECTIVE: With the current study, we aimed at describing our experience of surgical management of concealed penis using the modified Devine's procedure. PATIENTS AND METHODS: Medical records of all patients undergoing surgical management of concealed penis using the modified Devine's procedure from January 2019 to June 2021 were retrieved. Penile length was measured before and after the procedure. Parental satisfaction on the penile size, morphology, voiding status and hygiene was measured using a 5-point Likert's scale. RESULTS: A total of 131 patients were included. The mean age of the patients was 8.56 ± 2.75 years. The mean BMI of the patients was 24.17 ± 1.78 kg/m2. The patients were followed up to a mean of 15 ± 3 months. The duration of the surgery was 89.93 ± 16.85 minutes. There was a statistically significant increase in penile length from 1.8 ± 0.64 cm to 5.02 ± 1.11 cm after the procedure (p < 0.01). There was a statistically significant improvement in satisfaction status of all domains at 6, 12 and 24 weeks as compared to pre-operative levels. CONCLUSIONS: The modified Devine's technique is a simple and effective surgical technique for management of concealed penis in children producing predictable results and excellent parental satisfaction. The low rate of complications and good cosmetic outcomes lend support to its use in clinical practice.

Micropatterns of cell adhesive proteins with poly(ethylene oxide)-block-Poly(4-vinylpyridine) diblock copolymer.

Control of cell shape and behavior through the micropattern technique by spatial immobilization of adhesive proteins on a surface has provided novel insights in several aspects of cell biology, such as tissue morphogenesis, cell growth and cell differentiation, and apoptosis. In this work, we present the use of poly(ethylene oxide-block-poly(4-vinylpyridine) (PEO-b-P4VP) as a non-adhesive background to construct micropatterns of cell adhesive proteins. In the method presented, PEO-b-P4VP is used for its antifouling properties and at the same time, as a photosensitive material to define the micropatterns. The irradiation of PEO-b-P4VP with a short wavelength UV light through photolithographic mask, causes the polymer to crosslink and immobilize in the areas exposed. In the areas non-exposed the polymer can be removed. These areas can be subsequent back filled with the adhesive protein of interest to produce the final micropatterned cell chips.

High-sensitivity nanoscale chemical imaging with hard x-ray nano-XANES.

Resolving chemical species at the nanoscale is of paramount importance to many scientific and technological developments across a broad spectrum of disciplines. Hard x-rays with excellent penetration power and high chemical sensitivity are suitable for speciation of heterogeneous (thick) materials. Here, we report nanoscale chemical speciation by combining scanning nanoprobe and fluorescence-yield x-ray absorption near-edge structure (nano-XANES). First, the resolving power of nano-XANES was demonstrated by mapping Fe(0) and Fe(III) states of a reference sample composed of stainless steel and hematite nanoparticles with 50-nm scanning steps. Nano-XANES was then used to study the trace secondary phases in lithium iron phosphate (LFP) particles. We observed individual Fe-phosphide nanoparticles in pristine LFP, whereas partially (de)lithiated particles showed Fe-phosphide nanonetworks. These findings shed light on the contradictory reports on Fe-phosphide morphology in the literature. Nano-XANES bridges the capability gap of spectromicroscopy methods and provides exciting research opportunities across multiple disciplines.

Study of magnetic anisotropy and magnetization reversal using the quadratic magnetooptical effect in epitaxial Co(x)Mn(y)Ge(z)(111) films.

Magnetic anisotropy, magnetization reversal and the magnetooptic Kerr effect in Co(x)Mn(y)Ge(z) have been studied over a range of compositions between 0 and 50 at.% of Ge and between 1 and 3 in the Co to Mn atomic ratio, including the Heusler alloy Co(2)MnGe. A strong quadratic magnetooptic Kerr effect has been observed within a narrow region of composition centered around the Co to Mn atomic ratio of 2, which has been used to probe and quantify the magnetic anisotropy and magnetization reversal of the system. The anisotropy is sixfold with a weak uniaxial component, and it exhibits sensitive dependence on composition, especially on the atomic ratio between Co and Mn. The magnetization reversal process is consistent with the single-domain Stoner-Wohlfarth model.

Trimetazidine hydrochloride as a new treatment for patients with peripheral vascular disease--an exploratory trial.

OBJECTIVE: To evaluate the therapeutic effect of trimetazidine hydrochloride in peripheral vascular disease patients who had Rutherford classification grade 2-3. PATIENTS AND METHODS: 72 patients with Rutherford classification grade 2-3 in peripheral vascular were recruited successfully, they were randomly assigned to control group (35 cases) and trimetazidine group (37 cases), patients in control group received conventional treatment and trimetazidine group received conventional treatment plus trimetazidine hydrochloride for 6 months. Their ankle brachial index (ABI), maximum walking distance, pain onset time and the maximum walking time were compared before and after the treatment. RESULTS: After 6 months' treatment, the ABI, maximum walking distance, pain onset time and the maximum walking time in two groups were both improved of when compared with before treatment (p < 0.05). The maximum walking distance, pain onset time and the maximum walking time in trimetazidine group were improved better than control (p < 0.05) while no evident improvement in ABI between the 2 groups (p > 0.05). CONCLUSIONS: Conventional therapy plus trimetazidine hydrochloride could significantly improve the clinical symptoms of patients with Rutherford classification Grade 2-3 in peripheral vascular.

Association between variants of IL-8 and IL-10 genes, and efficacy of transcatheter arterial chemoembolization and subsequent prognosis in patients with liver cancer.

OBJECTIVE: Our objective was to examine the association between single nucleotide polymorphisms of interleukin (IL)-8 (rs4073 and rs2227306) and IL-10 (rs1800871 and rs1800872) genes, and clinical effects of transcatheter arterial chemoembolization (TACE) and subsequent prognosis in patients with liver cancer. PATIENTS AND METHODS: 115 patients with liver cancer underwent TACE. Venous blood specimens were collected for genomic DNA extraction. The restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) analysis was used to detect the above variants of IL-8 and IL-10 genes. In addition, blood levels of alpha fetal protein (AFP) were quantified by radioimmunoassay. Patients were followed up to uncover the association of the above genotypes with treatment efficacy and survival. RESULTS: Patients with the homozygous genotype AA or homozygous genotype TT (respectively, -251 and +781 sites) of IL-8 gene, and wild-type genotype TT or homozygous genotype AA (respectively, -819 and -592 sites) of IL-10 gene showed the best effectiveness of TACE. Furthermore, these patients also exhibited the lowest AFP levels and the longest survival after the treatment. CONCLUSIONS: Clinical efficacy of TACE and patient survival in liver cancer are associated with specific variants of IL-8 and IL-10 genes.

Molecular cloning and transcriptional mapping of the mouse cellular retinoic acid-binding protein gene.

The gene encoding the mouse cellular retinoic acid-binding protein (CRABP) has been isolated from a mouse genomic library and its structure has been determined. This gene spans approximately 10.5 kb and consists of four exons encoding 24, 59, 38, and 16 amino acid residues, respectively. This gene structure is very similar to the structures of other related genes belonging to the same protein family such as the human cellular retinol-binding protein, the rat cellular retinol-binding protein II, the rat fatty acid-binding protein, and the mouse adipocyte P2 protein. The site for transcription initiation has been mapped to the 93rd nucleotide upstream from the translation initiation codon ATG using both primer extension and RNase protection assays. From the DNA sequence, the promoter of the CRABP gene resembles those found in the "housekeeping" genes in that it is very G/C rich, lacks a TATA box, and contains multiple copies of the sequence GGGCGG. The deduced amino acid sequence of the translated region is identical to the amino acid sequence of the known bovine CRABP, and the DNA sequence of the transcribed region from the mouse gene shows approximately 78% homology to that of the bovine cDNA.

Circadian change of dopaminergic neuron activity: effects of constant light and melatonin.

Twenty-four hour profiles of tuberoinfundibular (TI), nigrostriatal and mesolimbic dopaminergic (DA) neuronal activities were assessed in estrogen-primed ovariectomized rats using DOPAC and DOPA levels in terminal regions of DA neurons. Significant decreases in DOPAC and DOPA levels in the median eminence were observed at 17.00 and 21.00 h, which corresponded with higher serum prolactin levels. DOPAC or DOPA levels in the striatum and nucleus accumbens were, however, significantly higher during the dark (21.00-05.00 h) phase. In rats kept under conditions of continuous light, no late afternoon decline in median eminence DOPA was observed; this decline could be reinstated by repeated injections of melatonin between 18.00 and 01.30 h for 3 days. In summary, circadian rhythms of central DA neurons were shown and melatonin may play an entraining role.

Design and performance of a scanning ptychography microscope.

We have designed and constructed a dedicated instrument to perform ptychography measurements and characterization of multilayer Laue lenses nanofocusing optics. The design of the scanning microscope provides stability of components and minimal thermal drifts, requirements for nanometer scale spatial resolution measurements. We performed thorough laboratory characterization of the instrument in terms of resolution and thermal drifts with subsequent measurements at a synchrotron. We have successfully acquired and reconstructed ptychography data yielding 11 nm line focus.

Compact prototype apparatus for reducing the circle of confusion down to 40 nm for x-ray nanotomography.

We have constructed a compact prototype apparatus for active correction of circle of confusion during rotational motion. Our system combines fiber optic interferometry as a sensing element, the reference cylinder along with the nanopositioning system, and a robust correction algorithm. We demonstrate dynamic correction of run-out errors down to 40 nm; the resolution is limited by ambient environment and accuracy of correcting nanopositioners. Our approach provides a compact solution for in-vacuum scanning nanotomography x-ray experiments with a potential to reach sub-nm level of correction.

Performance and characterization of the prototype nm-scale spatial resolution scanning multilayer Laue lenses microscope.

Synchrotron based x-ray microscopy established itself as a prominent tool for noninvasive investigations in many areas of science and technology. Many facilities around the world routinely achieve sub-micrometer resolution with a few instruments capable of imaging with the spatial resolution better than 100 nm. With an ongoing effort to push the 2D/3D resolution down to 10 nm in the hard x-ray regime both fabrication of the nano-focusing optics and stability of a microscope become extremely challenging. In this work we present our approach to overcome technical challenges on the path towards high spatial resolution hard x-ray microscopy and demonstrate the performance of a scanning fluorescence microscope equipped with the multilayer Laue lenses focusing optics.

Imaging cells and sub-cellular structures with ultrahigh resolution full-field X-ray microscopy.

Our experimental results demonstrate that full-field hard-X-ray microscopy is finally able to investigate the internal structure of cells in tissues. This result was made possible by three main factors: the use of a coherent (synchrotron) source of X-rays, the exploitation of contrast mechanisms based on the real part of the refractive index and the magnification provided by high-resolution Fresnel zone-plate objectives. We specifically obtained high-quality microradiographs of human and mouse cells with 29 nm Rayleigh spatial resolution and verified that tomographic reconstruction could be implemented with a final resolution level suitable for subcellular features. We also demonstrated that a phase retrieval method based on a wave propagation algorithm could yield good subcellular images starting from a series of defocused microradiographs. The concluding discussion compares cellular and subcellular hard-X-ray microradiology with other techniques and evaluates its potential impact on biomedical research.

An EMT spectrum defines an anoikis-resistant and spheroidogenic intermediate mesenchymal state that is sensitive to e-cadherin restoration by a src-kinase inhibitor, saracatinib (AZD0530).

The phenotypic transformation of well-differentiated epithelial carcinoma into a mesenchymal-like state provides cancer cells with the ability to disseminate locally and to metastasise. Different degrees of epithelial-mesenchymal transition (EMT) have been found to occur in carcinomas from breast, colon and ovarian carcinoma (OC), among others. Numerous studies have focused on bona fide epithelial and mesenchymal states but rarely on intermediate states. In this study, we describe a model system for appraising the spectrum of EMT using 43 well-characterised OC cell lines. Phenotypic EMT characterisation reveals four subgroups: Epithelial, Intermediate E, Intermediate M and Mesenchymal, which represent different epithelial-mesenchymal compositions along the EMT spectrum. In cell-based EMT-related functional studies, OC cells harbouring an Intermediate M phenotype are characterised by high N-cadherin and ZEB1 expression and low E-cadherin and ERBB3/HER3 expression and are more anoikis-resistant and spheroidogenic. A specific Src-kinase inhibitor, Saracatinib (AZD0530), restores E-cadherin expression in Intermediate M cells in in vitro and in vivo models and abrogates spheroidogenesis. We show how a 33-gene EMT Signature can sub-classify an OC cohort into four EMT States correlating with progression-free survival (PFS). We conclude that the characterisation of intermediate EMT states provides a new approach to better define EMT. The concept of the EMT Spectrum allows the utilisation of EMT genes as predictive markers and the design and application of therapeutic targets for reversing EMT in a selective subgroup of patients.