Identification of a novel inhibitor (NSC 665564) of dihydroorotate dehydrogenase with a potency equivalent to brequinar

A novel inhibitor of dihydroorotate dehydrogenase (DHO-DH) has been discovered using data from the National Cancer Institute's in vitro drug screen. Upon analysis of cytotoxicity results from the sixty tumor cell lines used in this screen, the COMPARE program predicted that NSC 665564 was likely to have the same mechanism of inhibition as brequinar, a known potent inhibitor of DHO-DH. We validated this prediction experimentally using MOLT-4 lymphoblast and found the IC50 of brequinar (0.5 microM) and NSC 665564 (0.3 microM) were comparable and that this induced cytotoxicity was reversed by either uridine or cytidine. The enzyme target of NSC 665564 was shown to be identical to that of brequinar when incubation with each drug followed by a 1 h pulse with [14C] sodium bicarbonate resulted in cellular accumulation of [14C]N-carbamyl-L-aspartic acid and [14C]L-dihydroorotic acid, with concurrent marked depletion of CTP and UTP. The Ki's for NSC 665564 and brequinar were 0.14 and 0.24 microM, respectively, when partially purified MOLT-4 mitochondria (the site of DHO-DH) were used. These results show that mechanistic predictions obtained using correlations from the COMPARE algorithm are independent of structure since the structure of NSC 665564 is dissimilar to that of other established DHO-DH inhibitors.     

Are toxicological responses in laboratory (inbred) zebrafish representative of those in outbred (wild) populations? - A case study with an endocrine disrupting chemical

Laboratory animals tend to be more inbred and less genetically diverse than wild populations, and thus may differ in their susceptibility to chemical stressors. We tested this hypothesis by comparing the responses of related inbred (theoretical inbreeding F(IT) = n + 0.25) and outbred (F(IT) = n) zebrafish (Danio rerio) WIK/Wild family lines to an endocrine disrupting chemical, clotrimazole. Exposure of inbred and outbred zebrafish to 2.9 μg clotrimazole/L had no effect on survival, growth, or gonadal development. Exposure of both lines to 43.7 μg clotrimazole/L led to male-biased sex ratios compared with controls (87% versus 55% and 92% vs 64%, for inbred and outbred males, respectively), advanced germ cell development, and reduced plasma 11-ketotestosterone concentrations in males. However, outbred males (but not inbred males) developed testis that were more than twice the weight of controls, which corresponded with a proliferation of Leydig cells and maintenance of the expression (rather than down-regulation occurring in inbreds) of gonadal aromatase (cyp19a1a) and insulin-like growth factor (igf1). Our results illustrate that the effects of an endocrine disrupting chemical (clotrimazole) on some end points (here testis development) can differ between inbred and outbred zebrafish. This highlights the need for reporting pedigree/genetic information and consistency in the responses of laboratory animals (e.g., by using model compounds as positive controls).     

2-(4-Aminophenyl)benzothiazoles: novel agents with selective profiles of in vitro anti-tumour activity

OBJECTIVE: To evaluate the results of cervical cytology screening in the National Breast and Cervical Cancer Early Detection Program and to compare the findings with results from other screening programs. METHODS: We analyzed data on 312,858 women aged 18 years and older who received one or more Papanicolaou smears, and follow-up if indicated, from October 1991 through June 1995 at screening sites across the United States providing comprehensive National Breast and Cervical Cancer Early Detection Program services. RESULTS: Of the women screened, more than half were 40 years or older; slightly less than half (44%) were of racial and ethnic minorities. During the first screening cycle, 3.8% of Papanicolaou tests were reported as abnormal (squamous intraepithelial lesion [SIL] or squamous cell cancer); proportions of abnormals decreased with increasing age. The age-adjusted rate of biopsy-confirmed cervical intraepithelial neoplasia (CIN) II or worse among women screened was 7.4 per 1000 Papanicolaou tests; rates of CIN were highest among young women, but cancer rates peaked among women in their 50s and 60s. The percentages of first screening cycle-Papanicolaou tests interpreted as high-grade SIL and squamous cell carcinoma associated with biopsy-confirmed CIN II or worse (the positive predictive value) were 56.0% for CIN II/III and 3.7% for invasive cancer. Of the 150 invasive cancers diagnosed, 54.0% were classified as local disease. CONCLUSION: Observed results emphasize the duality of cervical neoplasia-CIN in younger women and invasive cancer in older women. This finding points to the importance of reaching both younger and older women for cervical cancer screening.     

Cell-free V(D)J recombination

V(D)J recombination generates diversity in the immune system through the lymphoid-specific assembly of multiple gene segments into functional immunoglobulin and T-cell receptor genes. The first step in V(D)J recombination is cleavage of DNA at recombination signal sequences. Cleavage produces a blunt DNA end on each signal sequence and a hairpin end on adjacent coding gene segments, and can be reproduced in vitro by using purified RAG and RAG2 proteins. The later steps involve processing and joining of the cleaved DNA ends, and until now have been studied only in cells. Here we reconstitute the complete V(D)J recombination reaction in a cell-free system. We find that the RAG proteins are not only involved in cleavage, but are also needed in the later steps for efficient joining of coding ends. Joining is largely directed by short pieces of identical sequence in the coding flanks, but addition of human DNA ligase I results in greater diversity. Coding junctions contain short deletions as well as additions complementary to a coding flank (P nucleotides). Addition of non-templated nucleotides into coding junctions is mediated by terminal deoxyribonucleotidyl transferase. The cell-free reaction can therefore reproduce the complete set of processing events that occur in cells.     

Screening of elemental impurities in commercial detonation nanodiamond using sector field inductively coupled plasma-mass spectrometry

The quantitation of 55 elemental impurities in 15 commercial samples of detonation nanodiamond (DND), micron-sized diamond of high pressure/high temperature synthesis and detonation synthesis soot (DS) was achieved using a direct sector field ICP-MS analytical method. A set of 23 elements contributing more than 99.8 mass % of all impurities was selected and used as markers for the evaluation of DND purity. Obtained data were evaluated to identify important information on possible sources of nanodiamond contamination during purification, disaggregation, solubilisation or stabilisation of suspensions. Distinctive groups of elements (including Cr, Mn and S; Mo, W, V and Ti; Na, B and Si; Ca and Mg) occurring at elevated levels in DND can be readily visualised using radar plots, and can be related to the detonation synthesis (construction materials of explosion chamber, admixture in coolant, detonator type) and/or purification processes (type of oxidation process and reagents). The contaminant profile for each respective DND can be also considered as a fingerprint, characteristic for every producer and technology used. Results obtained also highlight how DND can be considered as effective collectors of various inorganic impurities from chemical reagents, glassware, sonotrode and other materials used during processing, including disaggregation and stabilisation of suspensions.     

Evaluating low-cost spectrometer designs for utility in reflectance and transmittance applications

Recent advances in low-cost microprocessors and sensors have allowed for the development of low-cost spectrometers for public experimentation and education outreach. In the context of the present paper and other recent research, low-cost spectrometers are important remote sensing instruments for numerous applications in citizen science and education. We built five spectrometer designs then tested them for absorption using concentrations of fluids and reflectance using colour swatches and minerals found on Mars as well as other materials possessing a high near infra-red (NIR) reflectance. These results were then compared with a laboratory grade spectrometer. We found that while all spectrometers produced meaningful results during the fluid concentration tests, there were considerable differences in accuracies between home-made instruments during the reflectance testing. Two spectrometer designs were identified that produced useful results for a range of low-cost scientific applications.     

Interfacial Strain Gradients Control Nanoscale Domain Morphology in Epitaxial BiFeO3 Multiferroic Films

Domain switching pathways fundamentally control performance in ferroelectric thin film devices. In epitaxial bismuth ferrite (BiFeO₃) films, the domain morphology is known to influence the multiferroic orders. While both striped and mosaic domains have been observed, the origins of the latter have remained unclear. Here, it is shown that domain morphology is defined by the strain profile across the film–substrate interface. In samples with mosaic domains, X‐ray diffraction analysis reveals strong strain gradients, while geometric phase analysis using scanning transmission electron microscopy finds that within 5 nm of the film–substrate interface, the out‐of‐plane strain shows an anomalous dip while the in‐plane strain is constant. Conversely, if uniform strain is maintained across the interface with zero strain gradient, striped domains are formed. Critically, an ex situ thermal treatment, which eliminates the interfacial strain gradient, converts the domains from mosaic to striped. The antiferromagnetic state of the BiFeO₃ is also influenced by the domain structure, whereby the mosaic domains disrupt the long‐range spin cycloid. This work demonstrates that atomic scale tuning of interfacial strain gradients is a powerful route to manipulate the global multiferroic orders in epitaxial films.     

Polarimetric Classification in a Tailings Deposition Area at the Timika Mine Site, Indonesia

Abstract.  Preliminary results indicate that polarimetric synthetic aperture radar (SAR) data can be useful in characterizing remote tailings sites in Indonesia. Recent launches of polarimetric radar satellites offer new opportunities to use such data. Wishart polarimetric radar classification was used to map tailings conditions and vegetative stress. We assessed the accuracy of our analysis and found that almost all scatterers could be discriminated properly, and that the technique was particularly useful in distinguishing saturated versus relatively dry tailings. Since tropical regions are subject to severe atmospheric disturbances, tailings stability is a major issue; there is thus high potential value in using radar remote sensing to aid revegetation and tailings stabilization.     

Controlled ultraviolet (UV) photoinitiated fabrication of monolithic porous layer open tubular (monoPLOT) capillary columns for chromatographic applications

An automated column fabrication technique that is based on a ultraviolet (UV) light-emitting diode (LED) array oven, and provides precisely controlled "in-capillary" ultraviolet (UV) initiated polymerization at 365 nm, is presented for the production of open tubular monolithic porous polymer layer capillary (monoPLOT) columns of varying length, inner diameter (ID), and porous layer thickness. The developed approach allows the preparation of columns of varying length, because of an automated capillary delivery approach, with precisely controlled and uniform layer thickness and monolith morphology, from controlled UV power and exposure time. The relationships between direct exposure times, intensity, and layer thickness were determined, as were the effects of capillary delivery rate (indirect exposure rate), and multiple exposures on the layer thickness and axial distribution. Layer thickness measurements were taken by scanning electron microscopy (SEM), with the longitudinal homogeneity of the stationary phase confirmed using scanning capacitively coupled contactless conductivity detection (sC(4)D). The new automated UV polymerization technique presented in this work allows the fabrication of monoPLOT columns with a very high column-to-column production reproducibility, displaying a longitudinal phase thickness variation within ±0.8% RSD (relative standard deviation).     

Processable Thermally Conductive Polyurethane Composite Fibers

The demand for wearable electronics has resulted in an increasing interest in the development of functional fibers, with a specific focus upon the development of electrically conductive fibers incorporable into garments. However, the production of thermally conductive fibers for heat dissipation has been largely neglected. Owing to the very rapid development of miniaturized wearable electronics, there is an increasing need for the development of thermally conductive fibers as heat sinks and thermal management processes. In this study, thermally conductive but electrically insulating boron nitride nanopowder (BNNP) fillers are used to effectively enhance the thermal conductivity and mechanical properties of elastomeric polyurethane fibers. Thermal conductivity enhancement of more than 160% is achieved at very low loadings of BNNP (less than 5 wt%) with an improvement in the mechanical properties of the unmodified fiber. These thermally conductive fibers are also incorporated into 3D textile structures as a proof of processability.