City of Palm Bay v. Bauman, 475 So. 2d 1322 (5th DCA 1985)

Constitutional Law-DRUG TESTING OF FLORIDA\u27S PUBLIC EMPLOYEES: WHEN MAY A PUBLIC EMPLOYER REQUIRE URINALYSIS

The Nature and Frequency of the Gas Outbursts in Comet 67P/Churyumov-Gerasimenko observed by the Alice Far-ultraviolet Spectrograph on Rosetta

Alice is a far-ultraviolet imaging spectrograph onboard Rosetta that, amongst multiple objectives, is designed to observe emissions from various atomic and molecular species from within the coma of comet 67P/Churyumov-Gerasimenko. The initial observations, made following orbit insertion in August 2014, showed emissions of atomic hydrogen and oxygen spatially localized close to the nucleus and attributed to photoelectron impact dissociation of H2O vapor. Weaker emissions from atomic carbon were subsequently detected and also attributed to electron impact dissociation, of CO2, the relative H I and C I line intensities reflecting the variation of CO2 to H2O column abundance along the line-of-sight through the coma. Beginning in mid-April 2015, Alice sporadically observed a number of outbursts above the sunward limb characterized by sudden increases in the atomic emissions, particularly the semi-forbidden O I 1356 multiplet, over a period of 10-30 minutes, without a corresponding enhancement in long wavelength solar reflected light characteristic of dust production. A large increase in the brightness ratio O I 1356/O I 1304 suggests O2 as the principal source of the additional gas. These outbursts do not correlate with any of the visible images of outbursts taken with either OSIRIS or the navigation camera. Beginning in June 2015 the nature of the Alice spectrum changed considerably with CO Fourth Positive band emission observed continuously, varying with pointing but otherwise fairly constant in time. However, CO does not appear to be a major driver of any of the observed outbursts.Comment: 6 pages, 4 figures, accepted for publication in the Astrophysical Journal Letter

Fataluku medicinal ethnobotany and the East Timorese military resistance

BACKGROUND: An ethnobotanical study of medicinal and poisonous plants used by the East Timor resistance was undertaken in the Lautem District of East Timor to study medicinal plant use in the region. Interviews were conducted with a single key consultant from the resistance army who belonged to the Fataluku culture. This study is of importance as a historical document and because no previous medicinal ethnobotanical studies on this region exist. METHODS: A rapid ethnobotanical survey of medicinal and poisonous plants was conducted through the proposed Conis Santana National Park in the Lautem district of East Timor. Medicinal and poisonous plants were identified by a Consultant and data was collected by the authors using classical descriptive ethnobotanical techniques (i.e. no quantitative measures) through an unstructured open ended interview. RESULTS: During the survey 40 medicinal and poisonous plants were identified by the Consultant and collected by the authors. Defining characteristics of the Consultant's knowledge include a high frequency use of trees, heavily forested habitats, leaves, decoctions and drinks for a range of conditions relevant to a resistance army. CONCLUSION: Despite limitations of the study, important contributions of this study include preservation of a part of the cultural history of the resistance movement and traditional botanical knowledge of the Fataluku. Furthermore, initial findings may indicate that traditional botanical knowledge is unique amongst different East Timorese cultures in terms of plant selection

A novel deconvolution method for modeling UDP-N-acetyl-D-glucosamine biosynthetic pathways based on 13C mass isotopologue profiles under non-steady-state conditions

<p>Abstract</p> <p>Background</p> <p>Stable isotope tracing is a powerful technique for following the fate of individual atoms through metabolic pathways. Measuring isotopic enrichment in metabolites provides quantitative insights into the biosynthetic network and enables flux analysis as a function of external perturbations. NMR and mass spectrometry are the techniques of choice for global profiling of stable isotope labeling patterns in cellular metabolites. However, meaningful biochemical interpretation of the labeling data requires both quantitative analysis and complex modeling. Here, we demonstrate a novel approach that involved acquiring and modeling the timecourses of ¹³C isotopologue data for UDP-<it>N</it>-acetyl-<smcaps>D</smcaps>-glucosamine (UDP-GlcNAc) synthesized from [U-¹³C]-glucose in human prostate cancer LnCaP-LN3 cells. UDP-GlcNAc is an activated building block for protein glycosylation, which is an important regulatory mechanism in the development of many prominent human diseases including cancer and diabetes.</p> <p>Results</p> <p>We utilized a stable isotope resolved metabolomics (SIRM) approach to determine the timecourse of ¹³C incorporation from [U-¹³C]-glucose into UDP-GlcNAc in LnCaP-LN3 cells. ¹³C Positional isotopomers and isotopologues of UDP-GlcNAc were determined by high resolution NMR and Fourier transform-ion cyclotron resonance-mass spectrometry. A novel simulated annealing/genetic algorithm, called 'Genetic Algorithm for Isotopologues in Metabolic Systems' (GAIMS) was developed to find the optimal solutions to a set of simultaneous equations that represent the isotopologue compositions, which is a mixture of isotopomer species. The best model was selected based on information theory. The output comprises the timecourse of the individual labeled species, which was deconvoluted into labeled metabolic units, namely glucose, ribose, acetyl and uracil. The performance of the algorithm was demonstrated by validating the computed fractional ¹³C enrichment in these subunits against experimental data. The reproducibility and robustness of the deconvolution were verified by replicate experiments, extensive statistical analyses, and cross-validation against NMR data.</p> <p>Conclusions</p> <p>This computational approach revealed the relative fluxes through the different biosynthetic pathways of UDP-GlcNAc, which comprises simultaneous sequential and parallel reactions, providing new insight into the regulation of UDP-GlcNAc levels and <it>O</it>-linked protein glycosylation. This is the first such analysis of UDP-GlcNAc dynamics, and the approach is generally applicable to other complex metabolites comprising distinct metabolic subunits, where sufficient numbers of isotopologues can be unambiguously resolved and accurately measured.</p

Room temperature ionic liquids with two symmetric ions

Room temperature ionic liquids typically contain asymmetric organic cations. The asymmetry is thought to enhance disorder, thereby providing an entropic counter-balance to the strong, enthalpic, ionic interactions, and leading, therefore, to lower melting points. Unfortunately, the synthesis and purification of such asymmetric cations is typically more demanding. Here we introduce novel room temperature ionic liquids in which both cation and anion are formally symmetric. The chemical basis for this unprecedented behaviour is the incorporation of ether-containing side chains – which increase the configurational entropy – in the cation. Molecular dynamics simulations indicate that the ether-containing side chains transiently sample curled configurations. Our results contradict the long-standing paradigm that at least one asymmetric ion is required for ionic liquids to be molten at room temperature, and hence open up new and simpler design pathways for these remarkable materials

Rhabdomyosarcoma cells show an energy producing anabolic metabolic phenotype compared with primary myocytes

<p>Abstract</p> <p>Background</p> <p>The functional status of a cell is expressed in its metabolic activity. We have applied stable isotope tracing methods to determine the differences in metabolic pathways in proliferating Rhabdomysarcoma cells (Rh30) and human primary myocytes in culture. Uniformly ¹³C-labeled glucose was used as a source molecule to follow the incorporation of ¹³C into more than 40 marker metabolites using NMR and GC-MS. These include metabolites that report on the activity of glycolysis, Krebs' cycle, pentose phosphate pathway and pyrimidine biosynthesis.</p> <p>Results</p> <p>The Rh30 cells proliferated faster than the myocytes. Major differences in flux through glycolysis were evident from incorporation of label into secreted lactate, which accounts for a substantial fraction of the glucose carbon utilized by the cells. Krebs' cycle activity as determined by ¹³C isotopomer distributions in glutamate, aspartate, malate and pyrimidine rings was considerably higher in the cancer cells than in the primary myocytes. Large differences were also evident in de novo biosynthesis of riboses in the free nucleotide pools, as well as entry of glucose carbon into the pyrimidine rings in the free nucleotide pool. Specific labeling patterns in these metabolites show the increased importance of anaplerotic reactions in the cancer cells to maintain the high demand for anabolic and energy metabolism compared with the slower growing primary myocytes. Serum-stimulated Rh30 cells showed higher degrees of labeling than serum starved cells, but they retained their characteristic anabolic metabolism profile. The myocytes showed evidence of de novo synthesis of glycogen, which was absent in the Rh30 cells.</p> <p>Conclusion</p> <p>The specific ¹³C isotopomer patterns showed that the major difference between the transformed and the primary cells is the shift from energy and maintenance metabolism in the myocytes toward increased energy and anabolic metabolism for proliferation in the Rh30 cells. The data further show that the mitochondria remain functional in Krebs' cycle activity and respiratory electron transfer that enables continued accelerated glycolysis. This may be a common adaptive strategy in cancer cells.</p

Exploring changing attitudes to non-invasive liver fibrosis tests in secondary care pathways: comparison of two national surveys

INTRODUCTION: The increasing availability of non-invasive tests (NITs) has created the opportunity to explore their use in improving risk stratification of advanced liver disease. The study aimed to determine the attitudes and practices among UK secondary care specialists, focusing primarily on attitudes to fibrosis assessment and the use of NITs. METHODS: Two web-based surveys were circulated, first between 2014 and 2015 (survey 1), and again in 2021 (survey 2). The surveys were promoted via the British Society of Gastroenterology, the British Association for the Study of the Liver and using Twitter. RESULTS: In survey 1, 215 healthcare professionals (HCPs) completed the online survey. 112 HCPs completed survey 2. 71 acute UK trusts were represented in survey 1 compared with 60 trusts in survey 2. Between the two surveys, the proportion of HCPs performing fibrosis assessment in all or nearly all cases rose from 45.1% to 74.1% (χ2=25.01; p&lt;0.0001). 46.5% (n=33/71) respondents in acute services reported the use of NITs in clinical pathways in survey 1, rising to 70.0% (n=42/60) in survey 2 (χ2=7.35; p=0.007). Availability of tests has increased but is not universal. The proportion reporting availability as a barrier to uptake fell from 57.2% of responses in survey 1 to 38.4% in 2021 χ2=11.01; p=0.0009). CONCLUSION: Between 2014 and 2021, the role of NITs in fibrosis assessment has risen substantially, as has the proportion of clinicians using NITs in clinical pathways to assess risk of liver disease. Poor access to NITs remains the predominant barrier

Altered regulation of metabolic pathways in human lung cancer discerned by 13C stable isotope-resolved metabolomics (SIRM)

<p>Abstract</p> <p>Background</p> <p>Metabolic perturbations arising from malignant transformation have not been systematically characterized in human lung cancers <it>in situ</it>. Stable isotope resolved metabolomic analysis (SIRM) enables functional analysis of gene dysregulations in lung cancer. To this purpose, metabolic changes were investigated by infusing uniformly labeled ¹³C-glucose into human lung cancer patients, followed by resection and processing of paired non-cancerous lung and non small cell carcinoma tissues. NMR and GC-MS were used for ¹³C-isotopomer-based metabolomic analysis of the extracts of tissues and blood plasma.</p> <p>Results</p> <p>Many primary metabolites were consistently found at higher levels in lung cancer tissues than their surrounding non-cancerous tissues. ¹³C-enrichment in lactate, Ala, succinate, Glu, Asp, and citrate was also higher in the tumors, suggesting more active glycolysis and Krebs cycle in the tumor tissues. Particularly notable were the enhanced production of the Asp isotopomer with three ¹³C-labeled carbons and the buildup of ¹³C-2,3-Glu isotopomer in lung tumor tissues. This is consistent with the transformations of glucose into Asp or Glu via glycolysis, anaplerotic pyruvate carboxylation (PC), and the Krebs cycle. PC activation in tumor tissues was also shown by an increased level of pyruvate carboxylase mRNA and protein.</p> <p>Conclusion</p> <p>PC activation – revealed here for the first time in human subjects – may be important for replenishing the Krebs cycle intermediates which can be diverted to lipid, protein, and nucleic acid biosynthesis to fulfill the high anabolic demands for growth in lung tumor tissues. We hypothesize that this is an important event in non-small cell lung cancer and possibly in other tumor development.</p

A dosimetric analysis of respiration-gated radiotherapy in patients with stage III lung cancer

BACKGROUND: Respiration-gated radiotherapy can permit the irradiation of smaller target volumes. 4DCT scans performed for routine treatment were retrospectively analyzed to establish the benefits of gating in stage III non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: Gross tumor volumes (GTVs) were contoured in all 10 respiratory phases of a 4DCT scan in 15 patients with stage III NSCLC. Treatment planning was performed using different planning target volumes (PTVs), namely: (i) PTV(routine), derived from a single GTV plus 'conventional' margins; (ii) PTV(all phases )incorporating all 3D mobility captured by the 4DCT; (iii) PTV(gating), incorporating residual 3D mobility in 3–4 phases at end-expiration. Mixed effect models were constructed in order to estimate the reductions in risk of lung toxicity for the different PTVs. RESULTS: Individual GTVs ranged from 41.5 – 235.0 cm(3). With patient-specific mobility data (PTV(all phases)), smaller PTVs were derived than when 'standard' conventional margins were used (p < 0.001). The average residual 3D tumor mobility within the gating window was 4.0 ± 3.5 mm, which was 5.5 mm less than non-gated tumor mobility (p < 0.001). The reductions in mean lung dose were 9.7% and 4.9%, respectively, for PTV(all phases )versus PTV(routine), and PTV(gating )versus PTV(all phases). The corresponding reductions in V(20 )were 9.8% and 7.0%, respectively. Dosimetric gains were smaller for primary tumors of the upper lobe versus other locations (p = 0.02). Respiratory gating also reduced the risks of radiation-induced esophagitis. CONCLUSION: Respiration-gated radiotherapy can reduce the risk of pulmonary toxicity but the benefits are particularly evident for tumors of the middle and lower lobes