A novel cell line panel reveals non-genetic mediators of platinum resistance and phenotypic diversity in high grade serous ovarian cancer.

OBJECTIVES: Resistance to cancer therapy is an enduring challenge and accurate and reliable preclinical models are lacking. We interrogated this unmet need using high grade serous ovarian cancer (HGSC) as a disease model. METHODS: We created five in vitro and two in vivo platinum-resistant HGSC models and characterised the entire cell panel via whole genome sequencing, RNASeq and creation of intraperitoneal models. RESULTS: Mutational signature analysis indicated that platinum-resistant cell lines evolved from a pre-existing ancestral clone but a unifying mutational cause for drug resistance was not identified. However, cisplatin-resistant and carboplatin-resistant cells evolved recurrent changes in gene expression that significantly overlapped with independent samples obtained from multiple patients with relapsed HGSC. Gene Ontology Biological Pathways (GOBP) related to the tumour microenvironment, particularly the extracellular matrix, were repeatedly enriched in cisplatin-resistant cells, carboplatin-resistant cells and also in human resistant/refractory samples. The majority of significantly over-represented GOBP however, evolved uniquely in either cisplatin- or carboplatin-resistant cell lines resulting in diverse intraperitoneal behaviours that reflect different clinical manifestations of relapsed human HGSC. CONCLUSIONS: Our clinically relevant and usable models reveal a key role for non-genetic factors in the evolution of chemotherapy resistance. Biological pathways relevant to the extracellular matrix were repeatedly expressed by resistant cancer cells in multiple settings. This suggests that recurrent gene expression changes provide a fitness advantage during platinum therapy and also that cancer cell-intrinsic mechanisms influence the tumour microenvironment during the evolution of drug resistance. Candidate genes and pathways identified here could reveal therapeutic opportunities in platinum-resistant HGSC.

Synthesis of Self-Assembled Randomly Oriented VO2 Nanowires on a Glass Substrate by a Spin Coating Method.

Randomly oriented vanadium dioxide (VO2) nanowires were produced on a glass substrate by spin coating from a cosolvent. SEM studies reveal that highly dense VO2 nanowires were grown at an annealing temperature of 400 °C. X-ray diffraction (XRD) provides evidence of the high crystallinity of the VO2 nanowires-embedded VO2 thin films on the glass substrate at 400 °C. Characterization by high-resolution transmission electron microscopy (HR-TEM) confirmed the formation of VO2 nanowires. The optical band gap of the nanowires-embedded VO2 thin films was also calculated from the transmittance data to be 2.65-2.70 eV. The growth mechanism of the solution-processed semiconducting VO2 nanowires was proposed based on both solvent selection and annealing temperature. Finally, the solar water splitting ability of the VO2 nanowires-embedded VO2 thin films was demonstrated in a photoelectrochemical cell (PEC).

Hydrothermal Synthesis and Crystal Structure of a (Ba0.54K0.46)4Bi4O12 Double-Perovskite Superconductor with Onset of the Transition Tc ∼ 30 K.

A new superconducting double perovskite was successfully synthesized by a low-temperature hydrothermal reaction at 240 °C. The crystal structure refinement of this double perovskite was done by single-crystal X-ray diffraction, and it had a cubic unit cell of a = 8.5207(2) Å with space group Im3̅m (No. 229). This superconducting double-perovskite chemical composition was estimated by electron probe microanalysis and was similar to the refined data. The superconducting transition temperature of the double perovskite was ∼30 K; the electrical resistivity began to fall at ∼25 K, and zero resistivity occurred below 7 K. Moreover, temperature-dependent resistivity under various magnetic fields and isothermal magnetization measurements ensured the nature of a type II superconductor for the sample. Finally, the metallic nature of the material was investigated by a first-principles study.

Hydrothermal Synthesis, Structure, and Superconductivity of Simple Cubic Perovskite (Ba0.62K0.38)(Bi0.92Mg0.08)O3 with Tc ∼ 30 K.

We have synthesized a new superconducting perovskite bismuth oxide by a facile hydrothermal route at 220 °C. The choice of starting materials, their mixing ratios, and the hydrothermal reaction temperature was crucial for obtaining products with superior superconducting properties. The structure of the powder sample was investigated using laboratory X-ray diffraction, high-resolution synchrotron X-ray diffraction (SXRD) data, and electron diffraction (ED) patterns [transmission electron microscopy (TEM) analysis]. The refinement of SXRD data confirmed a simple perovskite-type structure with a cubic cell of a = 4.27864(2) Å [space group Pm3̅m (No. 221)]. Elemental analysis detected magnesium in the final products, and a refinement based on SXRD and inductively coupled plasma data yielded an ideal undistorted simple cubic perovskite-type structure, with the chemical composition (Ba0.62K0.38)(Bi0.92Mg0.08)O3. ED patterns also confirmed the simple cubic perovskite structure; the cube-shaped microstructures and compositional homogeneity on the nanoscale were verified by scanning electron microscopy and TEM analyses, respectively. The fabricated compound exhibited a large shielding volume fraction of about 98% with a maximum Tcmag of ∼30 K, which was supported by the measured bismuth valence as well. Its electrical resistivity dropped at ∼21 K, and zero resistivity was observed below 7 K. The compound underwent thermal decomposition above 400 °C. Finally, the calculated band structure showed a metallic behavior for this hydrothermally synthesized bismuth oxide.

Superconducting double perovskite bismuth oxide prepared by a low-temperature hydrothermal reaction.

Perovskite-type structures (ABO3) have received significant attention because of their crystallographic aspects and physical properties, but there has been no clear evidence of a superconductor with a double-perovskite-type structure, whose different elements occupy A and/or B sites in ordered ways. In this report, hydrothermal synthesis at 220 °C produced a new superconductor with an A-site-ordered double perovskite structure, (Na(0.25)K(0.45))(Ba(1.00))3(Bi(1.00))4O12, with a maximum T(c) of about 27 K.

Scrutinizing transport phenomena and recombination mechanisms in thin film Sb2S3 solar cells.

The Schockley-Quisser (SQ) limit of 28.64% is distant from the Sb2S3 solar cells' record power conversion efficiency (PCE), which is 8.00%. Such poor efficiency is mostly owing to substantial interface-induced recombination losses caused by defects at the interfaces and misaligned energy levels. The endeavor of this study is to investigate an efficient Sb2S3 solar cell structure via accurate analytical modeling. The proposed model considers different recombination mechanisms such as non-radiative recombination, Sb2S3/CdS interface recombination, Auger, SRH, tunneling-enhanced recombination, and their combined impact on solar cell performance. This model is verified against experimental work (Glass/ITO/CdS/Sb2S3/Au) where a good coincidence is achieved. Several parameters effects such as thickness, doping, electronic affinity, and bandgap are scrutinized. The effect of both bulk traps located in CdS and Sb2S3 on the electrical outputs of the solar cell is analyzed thoroughly. Besides, a deep insight into the effect of interfacial traps on solar cell figures of merits is gained through shedding light into their relation with carriers' minority lifetime, diffusion length, and surface recombination velocity. Our research findings illuminate that the primary contributors to Sb2S3 degradation are interfacial traps and series resistance. Furthermore, achieving optimal band alignment by fine-tuning the electron affinity of CdS to create a Spike-like conformation is crucial for enhancing the immunity of the device versus the interfacial traps. In our study, the optimized solar cell configuration (Glass/ITO/CdS/Sb2S3/Au) demonstrates remarkable performance, including a high short-circuit current (JSC) of 47.9 mA/cm2, an open-circuit voltage (VOC) of 1.16 V, a fill factor (FF) of 54%, and a notable improvement in conversion efficiency by approximately 30% compared to conventional solar cells. Beyond its superior performance, the optimized Sb2S3 solar cell also exhibits enhanced reliability in mitigating interfacial traps at the CdS/Sb2S3 junction. This improved reliability can be attributed to our precise control of band alignment and the fine-tuning of influencing parameters.

Structural Engineering and Coupling of Two-Dimensional Transition Metal Compounds for Micro-Supercapacitor Electrodes.

The development of portable, wearable, and miniaturized integrated electronics has significantly promoted the immense desire for planar micro-supercapacitors (MSCs) among the extremely competitive energy storage devices. However, their energy density is still insufficient owing to the low electrochemical performance of conventional electrode materials. Compared with their bulk counterparts, the large specific surface area and fast ion transport with efficient intercalation of two-dimensional (2D) transition metal compounds have spurred the research platforms for their exploitation in the creation of high-performance MSCs. This Outlook presents a systematic summary of cutting-edge research on atomically thin, layered structures of transition metal dichalcogenides, MXenes, and transition metal oxides/hydroxides. Special emphasis is given to the rapid and durable storage of ions, benefiting from the low ion diffusion barriers of host interlayer spaces. Moreover, various strategies have been described to circumvent the structural damage due to the volume change and simultaneously evincing remarkable electronic properties.

Prevalence and Correlates of Depressive Symptoms among Medical Students: a Cross-sectional Single-centre Study.

BACKGROUND: Depressive symptoms are common among medical students. The aim of the present study was to determine the prevalence and risk factors of depressive symptoms among medical students in Sultan Qaboos University in Oman. METHOD: A cross-sectional study was conducted among a random sample selected from 1041 medical students at Sultan Qaboos University, Oman. The Patient Health Questionnaire-9 (PHQ-9) was used to screen for depressive symptoms. A logistic regression model was used to determine risk factors for depressive symptoms. RESULTS: Of 197 medical students selected, 189 (61 men and 128 women) responded. The PHQ-9 results showed that the prevalence of depressive symptoms was 41.3%. In multivariate analysis, female students were more likely than male students to develop depression (adjusted odds ratio = 2.866, p = 0.004). Medical students with a family history of depression were more likely to develop depression than those without a family history of depression (adjusted odds ratio = 4.150, p = 0.014). CONCLUSIONS: Depressive symptoms are common among medical students in Sultan Qaboos University. Risk factors for depressive symptoms are female sex and family history of depression.

The proteinase activated receptor-2 (PAR-2) mediates mitogenic responses in human vascular endothelial cells.

Proteolytically cleaved receptors, typified by the functional thrombin receptor (TR), represent a novel class of receptors that mediate signaling events by functional coupling to G proteins. Northern blot analysis completed with a human proteinase activated receptor-2 (PAR-2) cDNA as probe demonstrated the approximately 3.5kb PAR-2 transcript in total cellular RNA from human umbilical vein endothelial cells (HUVEC). Microspectrofluorimetry using Fura2-loaded HUVEC demonstrated a dose-dependent elevation in intracellular calcium transients ([Ca2+]i) to murine PAR39-44 (SLIGRL, putative neoligand after cleavage), with an approximate EC50 of 30 microM, and evidence for homologous desensitization with complete recovery at 45 min. Xenopus oocytes microinjected with TR cRNA failed to respond to 200 microM PAR39-44, and TR-targeted antisense oligonucleotides specifically abrogated thrombin-induced but not PAR39-44-mediated [Ca2+]i, excluding the possibility that TR/PAR-2 cell-surface coexpression was structurally linked. HUVEC incubated with PAR39-44 demonstrated a dose- and time-dependent mitogenic response similar to that seen with thrombin or TR42-47 (TR-activating peptide, SFLLRN). Preactivation of HUVEC with either PAR39-44 or thrombin resulted in heterologous desensitization to the corresponding agonist, an effect that was mediated primarily by TR internalization as evaluated by immunofluorescence and quantitative ELISA. These results ascribe a previously unrecognized function to the PAR-2 receptor, imply that a natural enzyme agonist may circulate in plasma, and suggest the presence of an additional regulatory mechanism controlling receptor activation events in vascular endothelial cells.

A survey of new long-stay hospital patients in an Irish health board area.

OBJECTIVES: The aims of this study were to determine the number and rate of accumulation of new long-stay hospital patients in one of Ireland's eight health board areas, to describe their demographic and clinical features, and to assess their needs in relation to possible community placement. METHODS: Demographic and clinical information was obtained on all patients over age 17 who had been continuously hospitalized in area hospitals for more than one year and less than six years on the census day of March 1, 1992. The Community Placement Questionnaire was used to rate the patients' social functioning, problem behavior, physical disability, social contact, and needs for accommodation and day care. RESULTS: The survey identified 175 new long-stay patients, mainly middle aged to elderly. Schizophrenia was the most common psychiatric diagnosis. The bed occupancy rate for these patients was 14 per 100,000 population, and the annual accumulation rate was 2.3 per 100,000 population. CONCLUSIONS: New long-stay patients were chronically ill with significant psychiatric and social disabilities. Involuntary patients were overrepresented in the group. Two-thirds could be placed in the community if facilities were available and had sufficiently high staffing levels.

Complex regional pain syndrome (CRPS) with resistance to local anesthetic block: a case report.

We present a case of complex regional pain syndrome (CRPS) Type 1 in a 12-year-old girl. The patient did not respond to the usual therapeutic modalities used to treat CRPS, including physical therapy, lumbar sympathetic block, epidural local anesthetic block, intravenous lidocaine infusion, or other oral medications. Of note is the fact that, during epidural block, the patient demonstrated a resistance to local anesthetic neural blockade in the area of the body involved with the pain problem. The mechanism of this resistance could be related to the changes in the dorsal horn cells of the spinal cord, secondary to activation of N-methyl-D-aspartate receptors, which may play a role in the pathophysiology of this pain syndrome.

The standardization of the Denver Developmental Screening Test on Arab children from the Middle East and north Africa.

A total of 936 children of Arab ethnic origin and culture were tested for the purpose of the standardization of the Denver Developmental Screening Test (DDST) on children from the Middle East and North Africa. There were 457 males and 479 females included in the study, with a race distribution of 216 white, 96 black, and 624 of mixed racial origin. The sample was divided into three age groups, with the age-range all-inclusive being from birth to six years. Five social classes were included. Accordingly, a new DDST screening form was designed and presented for the population studied. Age norms of developmental milestones on the personal-social, fine motor-adaptive, language, and gross motor skills are presented here.

The VLA-2 (alpha 2 beta 1) I domain functions as a ligand-specific recognition sequence for endothelial cell attachment and spreading: molecular and functional characterization.

The integrin VLA-2 (alpha 2 beta 1), generally considered to represent the specific collagen receptor on human endothelial cells, contains an alpha 2-subunit inserted I domain with structural similarity to the type A domains found within the recently described superfamily of receptor-ligand recognition proteins. This region of the cDNA has now been isolated and used for molecular and functional characterization of this heterodimeric receptor complex. Comparative sequence analysis with the porcine homologue revealed 93% amino acid sequence identity, suggestive of a developmentally conserved function. To complete structure/function studies, this region of the human cDNA was expressed as a chimeric protein in Escherichia coli, and a rabbit polyclonal antibody (anti-I domain) was used to study determinants of endothelial cell attachment and spreading in vitro. Quantifiable and visual disruption of endothelial cell attachment to gelatin, type I collagen, and laminin was evident using the specific anti-I domain antibody, with minimal inhibitory effects demonstrable using fibronectin or fibrinogen matrices. Therefore, these data would suggest that the alpha 2 beta 1 I domain confers ligand-binding specificity for both known alpha 2 beta 1 substrates (laminin and collagen), and that this region subserves a regulatory function in the molecular processes controlling endothelial cell attachment and spreading in vitro.

Conscious sedation with intermittent midazolam and fentanyl in electrophysiology procedures.

OBJECTIVES: To determine the safety and efficacy of intermittent midazolam and fentanyl conscious sedation for electrophysiology procedures (EP). BACKGROUND: Intermittent midazolam and fentanyl conscious sedation was administered in 700 consecutive cases (175 radiofrequency ablations, 163 EP studies, 261 pacemakers, and 101 implantable cardioverter-defibrillators) for 471 patients (239 males, 51%) mean age 65 +/- 15 years. The mean dose of midazolam was 0.063 mg/kg/hr and fentanyl was 0.591 microgram/kg/hr. METHODS: Cardiac rate and rhythm were monitored continuously, while blood pressure and arterial oxygen saturation were noninvasively assessed every 5 minutes. Drugs were administered in aliquots of 0.5 to 2.0 mg of midazolam and 6.25 to 25 micrograms of fentanyl as determined by clinical condition every 15 to 30 minutes. RESULTS: There were no deaths. In no case was endotracheal intubation required. Mild hypoxemia (SaO2 > 80%, but < 90%) occurred in 17 cases (2.4%) and was easily reversed with verbal stimulation and oropharyngeal repositioning (12 cases, 1.7%), increased F1O2 (3 cases, 0.4%), or intravenous naloxone (2 cases, 0.3%). Reversible hypotension (systolic blood pressure < 90, but > 60 mmHg) occurred in 14 patients (2.0%) and was corrected with intravenous crystalloid bolus or flumazenil (10 cases, 1.4%) or inotrope infusion (4 cases, 0.6%). No patient stay was prolonged due to sedation. Only five patients (0.7%) had any recollection of the procedure, while two (0.3%) were aware of pain. All hypoxemic episodes occurred during the first hour, whereas 43% (6/14) of hypotensive episodes occurred after the first hour. CONCLUSION: Conscious sedation with intermittent midazolam and fentanyl is safe and efficacious for a broad range of EP procedures.

Mitogenic responses mediated through the proteinase-activated receptor-2 are induced by expressed forms of mast cell alpha- or beta-tryptases.

The proteinase-activated receptor-2 (PAR-2) is the second member of a putative larger class of proteolytically activated receptors that mediate cell activation events by receptor cleavage or synthetic peptidomimetics corresponding to the newly generated N-terminus. To further study the previously identified mitogenic effects of PAR-2, we used the interleukin-3 (IL-3)-dependent murine lymphoid cell line, BaF3, for generation of stable cell lines expressing PAR-2 (BaF3/PAR-2) or the noncleavable PAR-2 mutant PAR-2(Arg36 --> Ala36). Only BaF3 cells expressing either wild-type or mutated receptor exhibited mitogenic responses when grown in IL-3-deficient media supplemented with PAR-2 activating peptide (SLIGRL, PAR39-44). This effect was dose dependent with an EC50 of approximately 80 micromol/L, sustained at 24, 48, and 72 hours, and was also demonstrable using thrombin receptor peptide TR42-47. Because tryptase shares approximately 70% homology with trypsin (previously shown to activate PAR-2), we studied recombinantly expressed forms of alpha- and beta-tryptases as candidate protease agonists for PAR-2. Hydrolytic activity of the chromogenic substrate tosyl-glycyl-prolyl-argly-4-nitroanilide acetate was present as a sharp peak at Mr approximately 130, confirming the presence of secretable and functionally active homotetrameric alpha- and beta-tryptases in transfected COS-1 cells. Dose-dependent proliferative responses were evident using either secreted form of tryptase with maximal responses seen at approximately 3 pmol/L (0.1 U/L). Receptor proteolysis was necessary and sufficient for mitogenesis because active site-blocked tryptase failed to induce this response, and proliferative responses were abrogated in BaF3 cells expressing PAR-2(Arg36 --> Ala36). These results specifically identify both forms of mast cell tryptases as serine protease agonists for PAR-2 and have implications for elucidating molecular mechanisms regulating cellular activation events mediated by proteases generated during inflammatory, fibrinolytic, or hemostatic-regulated pathways.

Vascular endothelial growth factor induces tissue factor and matrix metalloproteinase production in endothelial cells: conversion of prothrombin to thrombin results in progelatinase A activation and cell proliferation.

Production of vascular endothelial growth factor (VEGF) by cancer cells at invasive and metastatic sites is an important aspect of tumor angiogenesis. Although known primarily as a mitogen and a vascular permeability factor (VPF) for endothelial cells, VEGF/VPF has been proposed to induce the expression of procoagulant factors in endothelial cells. In this study, we have explored the ramifications of VEGF induction of tissue factor (TF) in human umbilical vein endothelial cells (HUVECs) and subsequent activation of progelatinase A. Within 3 hr of incubation with VEGF/VPF, endothelial cells accelerate TF generation as measured using chromogenic substrate assays for coagulation factors Xa and thrombin. Incubation of VEGF/VPF-pre-treated cells with prothrombin and factors X, Va, and VIIa at 37 degrees C and subsequent generation of thrombin resulted in activation of secreted endothelial progelatinase A as demonstrated by gelatin zymography. Anti-thrombin III or antibodies to TF inhibited thrombin generation and progelatinase A activation. VEGF/VPF also directly increased HUVEC secretion of interstitial collagenase, tissue inhibitor of metalloproteinases (TIMP-1) and, to a lesser extent, gelatinase A. The effect of thrombin on endothelial proliferation in serum-free media was examined. Thrombin was a growth factor for HUVECs at a lower dose than that required for progelatinase A activation. Whereas TIMP-2 abrogated thrombin-induced progelatinase A activation, it had no significant effect on thrombin-induced endothelial cell growth. We propose that an early step in tumor angiogenesis involves VEGF-induced thrombin generation and increased MMP production with subsequent activation of endothelial progelatinase A and degradation of the underlying basement membrane.