Women in support of non-violence against third world women

General note: Title and date provided by Bettye Lane.

"Pilado add Value" - valorisation of non-traditional marine resources

Os subprodutos da pesca têm recebido maior atenção devido à perceção dos seus impactos negativos na economia e no ambiente. Contudo, os subprodutos de crustáceos contêm vários compostos que podem ser usados como fonte de biopolímeros, como por exemplo a quitina, tendo esta uma grande variedade de aplicações biotecnológicas. O caranguejo Polybius henslowii, é um recurso marinho extremamente abundante na costa oeste Portuguesa durante os meses de Verão, não sendo contudo aproveitada para fins comerciais. Com este estudo procura-se assim contribuir para a valorização económica deste recurso, como fonte de matéria-prima para a extração de polímeros visando aplicações biotecnológicos. Para avaliar o seu potencial, a quitina foi extraída e quitosano produzido, a partir de distintas partes do corpo de Polybius henslowii: pereópodes e carapaça. O quitosano obtido, serviu, por sua vez, de matéria-prima para a produção de quitosano solúvel em água (WSC) e oligomeros de quitosano (COS), tendo todas as amostras sido caracterizadas e testadas quanto às suas propriedades antibacteriana, antifúngica e antioxidante, redução do radical 1, 1-Difenil-2-picrilhidrazil (DPPH). A caracterização das amostras de quitosano obtidas a partir de ambos os segmentos, demonstraram diferenças quanto ao seu rendimento, peso molecular e viscosidade. Os oligomeros de quitosano (COS) revelaram atividade antibacteriana contra todas as bactérias Gram-negativas testadas e a Gram-positiva, Lactobacillus planctarum (ATCC8014). COS mostraram melhores resultados na concentração mínima inibitória do que WSC contra todos os microrganismos testados, com maior inibição de crescimento demonstrada para Escherichia coli (ATCC10536) entre as concentrações de 0.125-0.0625 mg/mL. Os oligomeros demonstraram também maior atividade na redução do radical DPPH e na atividade antifúngica contra quatro espécies de fungo. A maior capacidade de redução foi obtida pelas amostras de COS obtidos a partir de pereópodes e carapaças (40%) com 1mg/mL de amostra.A capacidade de inibição do crescimento das espécies de fungos testados provou ser maior também para as amostras de oligomeros, do que para o quitosano solúvel em água. A maior atividade observada foi conseguida pelas amostras de pCOS (oligomeros de quitosano de pereópodes) e cCOS (oligomeros de quitosano de carapaças) para Cryphonectria parasitica (DSMZ 62626) com 84.5±3.14% e 85.6±2.27%, respetivamente. Tendo por base os resultados obtidos, é possível concluir pelas características bioquímicas e propriedades biológicas deste recurso marinho não tradicional, Polybius henslowii, suportam a sua utilização por indústrias biotecnológicas, promovendo assim a sua valorização económica. A exploração desta espécie vítima de captura acidental pode também aumentar desta forma a competitividade da atividade pesqueira através da reorientação e diversificação das espécies alvo, com potencial impacto no desenvolvimento sustentável nas comunidades costeiras.

Design of experiments to study the impact of process parameters on droplet size and development of non-invasive imaging techniques in tablet coating

Atomisation of an aqueous solution for tablet film coating is a complex process with multiple factors determining droplet formation and properties. The importance of droplet size for an efficient process and a high quality final product has been noted in the literature, with smaller droplets reported to produce smoother, more homogenous coatings whilst simultaneously avoiding the risk of damage through over-wetting of the tablet core. In this work the effect of droplet size on tablet film coat characteristics was investigated using X-ray microcomputed tomography (XμCT) and confocal laser scanning microscopy (CLSM). A quality by design approach utilising design of experiments (DOE) was used to optimise the conditions necessary for production of droplets at a small (20 μm) and large (70 μm) droplet size. Droplet size distribution was measured using real-time laser diffraction and the volume median diameter taken as a response. DOE yielded information on the relationship three critical process parameters: pump rate, atomisation pressure and coating-polymer concentration, had upon droplet size. The model generated was robust, scoring highly for model fit (R2 = 0.977), predictability (Q2 = 0.837), validity and reproducibility. Modelling confirmed that all parameters had either a linear or quadratic effect on droplet size and revealed an interaction between pump rate and atomisation pressure. Fluidised bed coating of tablet cores was performed with either small or large droplets followed by CLSM and XμCT imaging. Addition of commonly used contrast materials to the coating solution improved visualisation of the coating by XμCT, showing the coat as a discrete section of the overall tablet. Imaging provided qualitative and quantitative evidence revealing that smaller droplets formed thinner, more uniform and less porous film coats.

Dissecting the Functional Impacts of Non-Coding Genetic Variation

A large proportion of the variation in traits between individuals can be attributed to variation in the nucleotide sequence of the genome. The most commonly studied traits in human genetics are related to disease and disease susceptibility. Although scientists have identified genetic causes for over 4,000 monogenic diseases, the underlying mechanisms of many highly prevalent multifactorial inheritance disorders such as diabetes, obesity, and cardiovascular disease remain largely unknown. Identifying genetic mechanisms for complex traits has been challenging because most of the variants are located outside of protein-coding regions, and determining the effects of such non-coding variants remains difficult. In this dissertation, I evaluate the hypothesis that such non-coding variants contribute to human traits and diseases by altering the regulation of genes rather than the sequence of those genes. I will specifically focus on studies to determine the functional impacts of genetic variation associated with two related complex traits: gestational hyperglycemia and fetal adiposity. At the genomic locus associated with maternal hyperglycemia, we found that genetic variation in regulatory elements altered the expression of the HKDC1 gene. Furthermore, we demonstrated that HKDC1 phosphorylates glucose in vitro and in vivo, thus demonstrating that HKDC1 is a fifth human hexokinase gene. At the fetal-adiposity associated locus, we identified variants that likely alter VEPH1 expression in preadipocytes during differentiation. To make such studies of regulatory variation high-throughput and routine, we developed POP-STARR, a novel high throughput reporter assay that can empirically measure the effects of regulatory variants directly from patient DNA. By combining targeted genome capture technologies with STARR-seq, we assayed thousands of haplotypes from 760 individuals in a single experiment. We subsequently used POP-STARR to identify three key features of regulatory variants: that regulatory variants typically have weak effects on gene expression; that the effects of regulatory variants are often coordinated with respect to disease-risk, suggesting a general mechanism by which the weak effects can together have phenotypic impact; and that nucleotide transversions have larger impacts on enhancer activity than transitions. Together, the findings presented here demonstrate successful strategies for determining the regulatory mechanisms underlying genetic associations with human traits and diseases, and value of doing so for driving novel biological discovery.

Ocean acidification and its potential effects on the early life-history of non-calcifying and calcifying echinoderm (Odontaster validus, Patiriella regularis and Arachnoides placenta) larvae, 2011

Ocean acidification, as a result of increased atmospheric CO2, has the potential to adversely affect the larval stages of many marine organisms and hence have profound effects on marine ecosystems. This is the first study of its kind to investigate the effects of ocean acidification on the early life-history stages of three echinoderms species, two asteroids and one irregular echinoid. Potential latitudinal variations on the effects of ocean acidification were also investigated by selecting a polar species (Odontaster validus), a temperate species (Patiriella regularis), and a tropical species (Arachnoides placenta). The effects of reduced seawater pH levels on the fertilization of gametes, larval survival and morphometrics on the aforementioned species were evaluated under experimental conditions. The pH levels considered for this research include ambient seawater (pH 8.1 or pH 8.2), levels predicted for 2100 (pH 7.7 and pH 7.6) and the extreme pH of 7.0, adjusted by bubbling CO2 gas into filtered seawater. Fertilization for Odontaster validus and Patiriella regularis for the predicted scenarios for 2100 was robust, whereas fertilization was significantly reduced in Arachnoides placenta. Larval survival was robust for the three species at pH 7.8, but numbers declined when pH dropped below 7.6. Normal A. placenta larvae developed in pH 7.8, whereas smaller larvae were observed for O. validus and P. regularis under the same pH treatment. Seawater pH levels below 7.6 resulted in smaller and underdeveloped larvae for all three species. The greatest effects were expected for the Antarctic asteroid O. validus but overall the tropical sand dollar A. placenta was the most affected by the reduction in seawater pH. The effects of ocean acidification on the asteroids O. validus and P. regulars, and the sand dollar A. placenta are species-specific. Several parameters, such as taxonomic differences, physiology, genetic makeup and the population's evolutionary history may have contributed to this variability. This study highlights the vulnerability of the early developmental stages and the complexity of ocean acidification. However, future research is needed to understand the effects at individual, community and ecosystem levels.

Isolation and structural determination of non-racemic tertiary cathinone derivatives

The racemic tertiary cathinones N,N-dimethylcathinone (1), N,N-diethylcathinone (2) and 2-(1-pyrrolidinyl)-propiophenone (3) have been prepared in reasonable yield and characterized using NMR and mass spectroscopy. HPLC indicates that these compounds are isolated as the anticipated racemic mixture. These can then be co-crystallized with (+)-O,O′-di-p-toluoyl-d-tartaric, (+)-O,O′-dibenzoyl-d-tartaric and (-)-O,O′-dibenzoyl-l-tartaric acids giving the single enantiomers S and R respectively of 1, 2 and 3, in the presence of sodium hydroxide through a dynamic kinetic resolution. X-ray structural determination confirmed the enantioselectivity. The free amines could be obtained following basification and extraction. In methanol these are reasonably stable for the period of several hours, and their identity was confirmed by HPLC and CD spectroscopy.

Complete framework for efficient characterisation of non-Markovian processes

A non-Markovian process is one that retains `memory' of its past. A systematic understanding of these processes is necessary to fully describe and harness a vast range of complex phenomena; however, no such general characterisation currently exists. This long-standing problem has hindered advances in understanding physical, chemical and biological processes, where often dubious theoretical assumptions are made to render a dynamical description tractable. Moreover, the methods currently available to treat non-Markovian quantum dynamics are plagued with unphysical results, like non-positive dynamics. Here we develop an operational framework to characterise arbitrary non-Markovian quantum processes. We demonstrate the universality of our framework and how the characterisation can be rendered efficient, before formulating a necessary and sufficient condition for quantum Markov processes. Finally, we stress how our framework enables the actual systematic analysis of non-Markovian processes, the understanding of their typicality, and the development of new master equations for the effective description of memory-bearing open-system evolution.

Tyrosine kinase inhibitor insensitivity of non-cycling CD34+ human acute myeloid leukaemia cells with FMS-like tyrosine kinase 3 mutations.

The efficacy of tyrosine kinase (TK) inhibitors on non-cycling acute myeloid leukaemia (AML) cells, previously shown to have potent tumourigenic potential, is unknown. This pilot study describes the first attempt to characterize non-cycling cells from a small series of human FMS-like tyrosine kinase 3 (FLT3) mutation positive samples. CD34+ AML cells from patients with FLT3 mutation positive AML were cultured on murine stroma. In expansion cultures, non-cycling cells were found to retain CD34+ expression in contrast to dividing cells. Leukaemic gene rearrangements could be detected in non-cycling cells, indicating their leukaemic origin. Significantly, the FLT3-internal tandem duplication (ITD) mutation was found in the non-cycling fraction of four out of five cases. Exposure to the FLT3-directed inhibitor TKI258 clearly inhibited the growth of AML CD34+ cells in short-term cultures and colony-forming unit assays. Crucially, non-cycling cells were not eradicated, with the exception of one case, which exhibited exquisite sensitivity to the compound. Moreover, in longer-term cultures, TKI258-treated non-cycling cells showed no growth impairment compared to treatment-naive non-cycling cells. These findings suggest that non-cycling cells in AML may constitute a disease reservoir that is resistant to TK inhibition. Further studies with a larger sample size and other inhibitors are warranted.

Refining the diagnosis and EGFR status of non-small cell lung carcinoma in biopsy and cytologic material, using a panel of mucin staining, TTF-1, cytokeratin 5/6, and P63, and EGFR mutation analysis.

INTRODUCTION: The dichotomization of non-small cell carcinoma (NSCLC) subtype into squamous (SQCC) and adenocarcinoma (ADC) has become important in recent years and is increasingly required with regard to management. The aim of this study was to determine the utility of a panel of commercially available antibodies in refining the diagnosis on small biopsies and also to determine whether cytologic material is suitable for somatic EGFR genotyping in a prospectively analyzed series of patients undergoing investigation for suspected lung cancer. METHODS: Thirty-two consecutive cases of NSCLC were first tested using a panel comprising cytokeratin 5/6, P63, thyroid transcription factor-1, 34betaE12, and a D-PAS stain for mucin, to determine their value in refining diagnosis of NSCLC. After this test phase, two further pathologists independently reviewed the cases using a refined panel that excluded 34betaE12 because of its low specificity for SQCC, and refinement of diagnosis and concordance were assessed. Ten cases of ADC, including eight derived from cytologic samples, were sent for EGFR mutation analysis. RESULTS: There was refinement of diagnosis in 65% of cases of NSCLC to either SQCC or ADC in the test phase. This included 10 of 13 cases where cell pellets had been prepared from transbronchial needle aspirates. Validation by two further pathologists with varying expertise in lung pathology confirmed increased refinement and concordance of diagnosis. All samples were adequate for analysis, and they all showed a wild-type EGFR genotype. CONCLUSION: A panel comprising cytokeratin 5/6, P63, thyroid transcription factor-1, and a D-PAS stain for mucin increases diagnostic accuracy and agreement between pathologists when faced with refining a diagnosis of NSCLC to SQCC or ADC. These small samples, even cell pellets derived from transbronchial needle aspirates, seem to be adequate for EGFR mutation analysis.

A Loop-mediated Isothermal Amplification Method for Rapid Direct Detection and Differentiation of Non-pathogenic and Verocytotoxigenic Escherichia coli in Beef and Bovine Faeces

The aim of this study was to develop a multiplex loop-mediated isothermal amplification (LAMP) method capable of detecting Escherichia coli generally and verocytotoxigenic E. coli (VTEC) specifically in beef and bovine faeces. The LAMP assay developed was highly specific (100%) and able to distinguish between E. coli and VTEC based on the amplification of the phoA, and stx1 and/or stx2 genes, respectively. In the absence of an enrichment step, the limit of detection 50% (LOD50) of the LAMP assay was determined to be 2.83, 3.17 and 2.83-3.17 log CFU/g for E. coli with phoA, stx1 and stx2 genes, respectively, when artificially inoculated minced beef and bovine faeces were tested. The LAMP calibration curves generated with pure cultures, and spiked beef and faeces, suggested that the assay had good quantification capability. Validation of the assay, performed using retail beef and bovine faeces samples, demonstrated good correlation between counts obtained by the LAMP assay and by a conventional culture method, but suggested the possibility of false negative LAMP results for 12.5-14.7% of samples tested. The multiplex LAMP assay developed potentially represents a rapid alternative to culture for monitoring E.coli levels in beef or faeces and it would provide additional information on the presence of VTEC. However, some further optimisation is needed to improve detection sensitivity.

Conclusion: patterns of non-territorial autonomy

This article seeks to generalise about the significance of non-territorial autonomy as a mechanism for the management of ethnic conflict on the basis of a set of case studies covering the Ottoman empire and its successor states, the Habsburg monarchy, the Jewish minorities of Europe, interwar Estonia, contemporary Belgium, and two indigenous peoples, the Sami in Norway and Maori in New Zealand. It begins by assessing the extent to which the spatial distribution of ethnonational communities determined the range of autonomy options available—whether these might be territorial or whether only non-territorial autonomy would be realistic. The article continues with an assessment of the significance of ‘autonomy’ in circumstances where the institutions with which it is associated enjoy a non-territorial rather than a territorial writ. It concludes by suggesting that in almost all cases where autonomy is extended to a minority within a state this is exercised on a territorial basis, and that in many cases of non-territorial autonomy, or national–cultural autonomy, the powers assumed by the ‘autonomous’ institutions are substantially symbolic. It argues that notwithstanding the limited empirical evidence for the existence of non-territorial autonomy, this device should not be written off at a normative level.