The synthesis and properties of some hydrazines

A new synthetic method, applicable to the preparation of a wide range of hydrazine derivatives, is described. This involves the diborane reduction of a hydrazone, or, more conveniently, the reductive-condensation of a hydrazine and the appropriate aldehyde (or ketone). The method gives high yields and provides a particularly simple route to the relatively inaccessible 1,2-disubstituted hydrazines bearing a different group on each nitrogen. The new method has also been applied to the preparation of 1,2-disubstituted hydrazines with the same group on both nitrogens (via the azine), the very rare 1 ,2-disubstituted hydrazines bearing a tert-butyl group, trisubstituted hydrazines and monosubstituted hydrazines. Application of the reaction to the preparation of diaziridines has also been investigated. A mechanism for the reduction, supported by the isolation of a boron-containing intermediate, is suggested. Some limitations of the procedure are discussed. A general i.r. method of distinguishing the isomeric disubstituted hydrazines, as stable salts, has been developed. This has the advantages of speed and simplicity over previous methods. The mass spectra of a series of monosubstituted hydrazines, a series of 1,2-disubstituted hydrazines and some 1-benzoyl 2-alkylhydrazines have been examined in detail. The spectra are generally dominated byα -cleavage processes and the compounds show a variety of interesting rearrangement reactions. The mass spectra of some 1, 1-disubstituted hydrazines and some trisubstituted hydrazines have also been examined. Rearrangement processes occurring in the mass spectrum of tropylium fluoroborate have been examined. Similar rearrangements have been found in the spectrum of trityl fluoroborate and may be of general occurrence in the mass spectra of aromatic fluoroborates. Chemical shift values for some groups on hydrazine nitrogen are recorded and the results of tumour inhibitory tests on some hydrazines are also given.

FLQ, the Fastest Quadratic Complexity Bound on the Values of Positive Roots of Polynomials

In this paper we present F LQ, a quadratic complexity bound on the values of the positive roots of polynomials. This bound is an extension of FirstLambda, the corresponding linear complexity bound and, consequently, it is derived from Theorem 3 below. We have implemented FLQ in the Vincent-Akritas-Strzeboński Continued Fractions method (VAS-CF) for the isolation of real roots of polynomials and compared its behavior with that of the theoretically proven best bound, LM Q. Experimental results indicate that whereas F LQ runs on average faster (or quite faster) than LM Q, nonetheless the quality of the bounds computed by both is about the same; moreover, it was revealed that when VAS-CF is run on our benchmark polynomials using F LQ, LM Q and min(F LQ, LM Q) all three versions run equally well and, hence, it is inconclusive which one should be used in the VAS-CF method.

A novel role of plasma membrane calcium atpase 4 as a negative-regulator of VEGF-induced angiogenesis

Current anti-angiogenic treatments involve the attenuation of signalling via the pro-angiogenic vascular endothelial growth factor/receptor (VEGF/VEGFR) axis. Stimulation of angiogenesis by VEGF requires the activation of the calcineurin/nuclear factor of activated T-cells (NFAT) signal transduction pathway which is inhibited by Plasma Membrane Calcium ATPase 4 (PMCA4), an endogenous calcium extrusion pump. However, PMCA4s role in calcineurin/NFAT-dependent angiogenesis is unknown. Using “gain of function” studies, we show here that adenoviral overexpression of PMCA4 in human umbilical vein endothelial cells (HUVEC) inhibited NFAT activity, decreased the expression of NFAT-dependent pro-angiogenic proteins (regulator of calcineurin 1.4 (RCAN1.4) and cyclooxygenase-2) and diminished in vitro cell migration and tube formation in response to VEGF-stimulation. Furthermore, in vivo blood vessel formation was attenuated in a matrigel plug assay by ectopic expression of PMCA4. Conversely, “loss of function” experiments by si-RNA-mediated knockdown of PMCA4 in HUVEC or isolation of mouse lung endothelial cells from PMCA4−/− mice showed increased VEGF-induced NFAT activity, RCAN1.4 expression, in vitro endothelial cell migration, tube formation and in vivo blood vessel formation. Additionally, in an in vivo pathological angiogenesis model of limb ischemia, the reperfusion of the ischemic limb of PMCA4−/− mice was augmented compared to wild-type. Disruption of the interaction between endogenous PMCA4 and calcineurin by adenoviral overexpression of the region of PMCA4 that interacts with calcineurin (residues 428–651) increased NFAT activity, RCAN1.4 protein expression and in vitro tube formation. These results identify PMCA4 as an inhibitor of VEGF-induced angiogenesis, highlighting its potential as a new therapeutic target for anti-angiogenic treatments.

Examination of the role of Mycoplasma bovis in bovine pneumonia and a mathematical model for its evaluation

The authors screened 34 large cattle herds for the presence of Mycoplasma bovis infection by examining slaughtered cattle for macroscopic lung lesions, by culturing M. bovis from lung lesions and at the same time by testing sera for the presence of antibodies against M. bovis. Among the 595 cattle examined, 33.9% had pneumonic lesions, mycoplasmas were isolated from 59.9% of pneumonic lung samples, and 10.9% of sera from those animals contained antibodies to M.bovis. In 25.2% of the cases M. bovis was isolated from lungs with no macroscopic lesions. The proportion of seropositive herds was 64.7%. The average seropositivity rate of individuals was 11.3% but in certain herds it exceeded 50%. A probability model was developed for examining the relationship among the occurrence of pneumonia, the isolation of M. bovis from the lungs and the presence of M. bovis specific antibodies in sera.

Sequence analysis of complementary DNAs encoding C3 in the nurse shark ({\it Ginglymostoma cirratum\/})

Mammalian C3 is a pivotal complement protein, encoded for by a single gene. In some vertebrate species multiple C3 isoforms are products of different C3 genes. The goal of this study was to determine whether multiple genes encode for shark C3. A protocol was developed for the isolation of mRNA from shark blood for the isolation of C3 cDNA clones. RT-PCR amplification of mRNA, using sense (GCGEQNM) and antisense (TWLTAYV) primers encoding conserved regions of human C3, yielded 21 clones. The C3-like clones isolated shared 97% similarity with each other and 40% similarity to human C3. RACE-PCR amplification of shark liver RNA, using gene specific primers, yielded products ranging from 1800bp to 3000bp. Deduced amino acid sequence, corresponding to 408bp of the 1800bp fragment, was obtained which showed 51% similarity to human C3. These results suggest that nurse shark C3 might be encoded for by more than one gene. ^

Identification and localization of polyketide synthase genes from cultures of diarrheic shellfish poisoning toxin producing dinoflagellates of the genus Prorocentrum

Aquatic toxins are responsible for a number of acute and chronic diseases in humans. Okadaic acid (OA) and other dinoflagellate derived polyketide toxins pose serious health risks on a global scale. Ingestion of OA contaminated shellfish causes diarrheic shellfish poisoning (DSP). Some evidence also suggests tumor promotion in the liver by OA. Microcystin-LR (MC-LR) is produced by cyanobacteria and is believed to be the most common freshwater toxin in the US. Humans may be exposed to this acute hepatotoxin through drinking or recreational use of contaminated waters. ^ OA producing dinoflagellates have not been cultured axenically. The presence of associated bacteria raises questions about the ultimate source of OA. Identification of the toxin-producing organism(s) is the first step in identifying the biosynthetic pathways involved in toxin production. Polyketide synthase (PKS) genes of toxic and non-toxic species were surveyed by construction of clonal libraries from PCR amplicons of various toxic and non-toxic species of Prorocentrum in an effort to identify genes, which may be part of the biosynthetic pathway of OA. Analysis of the PKS sequences revealed that toxic species shared identical PKS genes not present in non-toxic species. Interestingly, the same PKS genes were identified in a library constructed from associated bacteria. ^ Subsequent bacterial small subunit RNA (16S) clonal libraries identified several common bacterial species. The most frequent 16S sequences found were identified as species of the genus Roseobacter which has previously been implicated in the production of OA. Attempts to culture commonly occurring bacteria resulted in the isolation of Oceanicaulis alexandrii , a novel marine bacterium previously isolated from the dinoflagellate Alexandrium tamarense, from both P. lima, and P. hoffmanianum. ^ Metabolic studies of microcystin-LR, were conducted to probe the activity of the major human liver cytochromes (CYP) towards the toxin. CYPs may provide alternate routes of detoxification of toxins when the usual routes have been inhibited. For example, some research indicates that cyanobacterial xenobiotics, in particular, lipopolysaccharides may inhibit glutathione S-transferases allowing the toxin to persist long enough to be acted upon by other enzymes. These studies found that at least one human liver CYP was capable of metabolizing the toxin. ^

The rise of Islamism among Turkish immigrants in Germany and the Netherlands

This study is a comparative study of the Turkish Islamist movement Milli Görüş (IGMG) in Germany and the Netherlands. It is a qualitative and a quantitative inquiry about the nature (whether it is moderate or radical) of the Milli Görüş movement in these two countries. The central research questions are: what is the reason for the rise of Islamism among Turks living in two different countries in Europe? What is the reason for the difference in the radicalization levels? Islamism refers to an ideology that turns traditional Islam into a sustained and systematic program that includes social, political, and economic affairs (Pipes 1998). The movements within the framework of Islamism range from moderate to radical. Based on the data collected during the field research conducted in Germany and the Netherlands between the years of 2004-2007, this study suggests that Islamism is a response to social marginalization which is defined as “an external social position, of isolation of the individual or groups, with a limited access to economical, political, educational and communicational resources of the community” (Contained in the law adopted by the Romanian Parliament in 28 February 2002, www.hurriyetim.com, November 25, 2004). It is hypothesized that as the level of social marginalization increases, so does the level of radicalization.

Scope & nature of young sudden cardiac death in Newfoundland & Labrador

Introduction: Sudden cardiac death (SCD) in young people (ages 2-40) is a tragedy for families and communities alike. It has multiple causes, one of which is an underlying genetic arrhythmogenic cardiomyopathy. A study from Ontario (ON) using a 2008 cohort assessed the incidence of SCD in persons aged 2-40 years to be 2.64/100,000 person-years. We hypothesized that Newfoundland & Labrador (NL) may have a higher incidence of early SCD in ages 2-40 due to possible underlying genetic causes given the historical genetic isolation of the population and the founder mutations already identified (ex. PKP2, RYR2, TMEM43). Methods: We ascertained cases of sudden death from the comprehensive Medical Examiners’ provincial database for the years 2008 and 1997; 2008 as a direct comparison to ON, and 1997 as it represented a time when the implantable cardioverter-defibrillator was not available in NL. Each case of sudden death was individually analyzed to determine likelihood of SCD. Results: There were 119 cases in 2008 and 157 cases in 1997. The incidence of SCD for ages 2-40 in 2008 was 7.32/100,000 persons. This was significantly higher than the incidence in Ontario. The incidence of SCD was not significantly higher in 1997 than 2008. Coronary artery disease was a major cause of death in all cohorts, similar to Ontario (non-significant difference). Conclusion: In general, there was a trend of more arrhythmogenic deaths in the young and more structural cardiac deaths as age increased. This reflects the cause of SCD in the young is often genetic in nature, while older deaths are often due to coronary artery disease, a disease heavily influenced by environment. To conclude, SCD in NL occurs at a higher incidence than ON, further research is needed on the topic.

Annotated record of the detailed examination of Mn deposits from COCOTOW Expedition stations in the intersection of the Siqueiros Transform Fault and the East Pacific Rise

A near-bottom geological and geophysical survey was conducted at the western intersection of the Siqueiros Transform Fault and the East Pacific Rise. Transform-fault shear appears to distort the east flank of the rise crest in an area north of the fracture zone. In ward-facing scarps trend 335° and do not parallel the regional axis of spreading. Small-scale scarps reveal a hummocky bathymetry. The center of spreading is not a central peak but rather a 20-40 m deep, 1 km wide valley superimposed upon an 8 km wide ridge-crest horst. Small-scale topography indicates widespread volcanic flows within the valley. Two 0.75 km wide blocks flank the central valley. Fault scarps are more dominant on the western flank. Their alignment shifts from directions intermediate to parallel to the regional axis of spreading (355°). A median ridge within the fracture zone has a fault-block topography similar to that of the East Pacific Rise to the north. Dominant eastward-facing scarps trending 335° are on the west flank. A central depression, 1 km wide and 30 m deep, separates the dominantly fault-block regime of the west from the smoother topography of the east flank. This ridge originated by uplift due to faulting as well as by volcanism. Detailed mapping was concentrated in a perched basin (Dante's Hole) at the intersection of the rise crest and the fracture zone. Structural features suggest that Dante's Hole is an area subject to extreme shear and tensional drag resulting from transition between non-rigid and rigid crustal behavior. Normal E-W crustal spreading is probably taking place well within the northern confines of the basin. Possible residual spreading of this isolated rise crest coupled with shear drag within the transform fault could explain the structural isolation of Dante's Hole from the remainder of the Siqueiros Transform Fault.

(Table 2) Age determination of Aegean Sea sediments

The modern Aegean Sea is an important source of deep water for the eastern Mediterranean. Its contribution to deep water ventilation is known to fluctuate in response to climatic variation on a decadal timescale. This study uses marine micropalaeontological and stable isotope data to investigate longer-term variability during the late glacial and Holocene, in particular that associated with the deposition of the early Holocene dysoxic/anoxic sapropel S1. Concentrating on the onset of sapropel-forming conditions, we identify the start of 'seasonal' stratification and highlight a lag in d18O response of the planktonic foraminifer N. pachyderma to termination T1b as identified in the d18O record of G. ruber. By use of a simple model we determine that this offset cannot be a function of bioturbation effects. The lag is of the order of 1 kyr and suggests that isolation of intermediate/deep water preceded the start of sapropel formation by up to 1.5 kyr. Using this discovery, we propose an explanation for the major unresolved problem in sapropel studies, namely, the source of nutrient supply required for export productivity to reach levels needed for sustained sapropel deposition. We suggest that nutrients had been accumulating in a stagnant basin for 1-1.5 kyr and that these accumulated resources were utilized during the deposition of S1. In addition, we provide a first quantitative estimate of the diffusive (1/e) mixing timescale for the eastern Mediterranean in its "stratified" sapropel mode, which is of the order of 450 years.

Morphology of Cycladophora davisiana davisiana from the Atlantic Ocean

In recent years, temporal fluctuations in the abundance of C. d. davisiana have been used frequently as a highresolution stratigraphic and paleoenvironmental tool. The modern ecology and morphologic variation (temporal and geographic) of this radiolarian species is evaluated to ascertain its potential stratigraphic and paleoenvironmental significance. Statistics were obtained on the width and height of all C. d. davisiana segments from Pleistocene populations of differing ages from the Northern Hemisphere (Labrador Sea and Iceland-Faeroe Ridge) and Southern Hemisphere (Namibian shelf and Meteor Rise). Results reveal that segment height variations between and within populations are more conservative than segment width. The mean sizes of the thorax and first abdominal segment have distinguishable differences between C. d. davisiana found in the North and South Atlantic. All populations have no significant difference in first abdominal segment width, however, mean heights of this segment differ greatly between populations of the North and South Atlantic. Second abdominal segment sizes show no clear population grouping. Size differences in post-cephalic segment size of these populations would appear to be related to some isolation of gene pools and possibly unknown paleoenvironmental factors. Temporal changes in the postcephalic size of C. d. davisiana may be used to: (1) identify temporally equivalent peaks in abundance of the species in a given region, (2) possibly evaluate the degree of mixing of water'masses between regions, and (3) trace the initial spread of the species from its area of origin. Cleve's 1887 plankton samples, between Greenland and Spitzsbergen, were studied and used in conjunction with other data to make the following conclusions on the modern ecology of C. d. davisiana in the Arctic and Greenland-Norwegian Seas. (1) It is presently absent in surface water plankton samples, (2) it currently lives at depths below 500 m, where it is rare, (3) it does not live in the upper 200 m under Arctic ice but is rare at greater depths, (4) it is absent in the upper 200 m near permanent Greenland Sea ice where normal oceanic salinity prevails, and (5) it is most common in deep marginal fjord environments which may serve as a refuge for the species during interglacial periods. In the Atlantic Ocean, the abundance of C. d. davisiana does not exceed 1% of the assemblage between the Subtropical Convergence of each hemisphere. In the Norwegian and Labrador Seas the species may occasionally be in the range of 1-5% of the modern radiolarian assemblage and never more than 5% in the southern high latitudes. Apparently only in the modern Sea of Okhotsk, does the species presently occur in high abundance. We concur with Morley and Hays (1983) that increased abundances are likely caused by the development of a strong low-salinity surface layer associated with seasonal sea ice melting and a strong temperature minimum above warmer and higher salinity intermediate waters. Similar conditions were frequent during the Pleistocene in the high latitudes and its modern scarcity outside the Sea of Okhotsk must be related to the absence of the presently unique conditions in the latter region.

(Table 1) Age determination of sediments from the Campbell Plateau

Campbell Plateau occupies a key position in the southwest Pacific sector of the Southern Ocean. The plateau confines and steers the Antarctic Circumpolar Current (ACC) along its flanks, isolating the Subantarctic plateau from cold polar waters. Oxygen and carbon isotope records from Campbell Plateau cores provide new records of water mass stratification for the past 130 kyr. During glacial climes, strengthening of the Subantarctic Front (SAF) caused waters over the plateau flanks to be deeply mixed and ~3°C cooler. Waters of the plateau interior remained stratified and isolated from the cold southern waters. In the west, waters cooled markedly (~4°C) owing to reduced entrainment of Tasman Sea water. Marked cooling also occurred north of Campbell Plateau under increased entrainment of polar water by a branch of the SAF. The ACC remained along the flanks of Campbell Plateau during the last interglacial, when interior waters were stratified and warmer by ~1°C than now.

CLARITY and PACT-based imaging of adult zebrafish and mouse for whole-animal analysis of infections.

Visualization of infection and the associated host response has been challenging in adult vertebrates. Owing to their transparency, zebrafish larvae have been used to directly observe infection in vivo; however, such larvae have not yet developed a functional adaptive immune system. Cells involved in adaptive immunity mature later and have therefore been difficult to access optically in intact animals. Thus, the study of many aspects of vertebrate infection requires dissection of adult organs or ex vivo isolation of immune cells. Recently, CLARITY and PACT (passive clarity technique) methodologies have enabled clearing and direct visualization of dissected organs. Here, we show that these techniques can be applied to image host-pathogen interactions directly in whole animals. CLARITY and PACT-based clearing of whole adult zebrafish and Mycobacterium tuberculosis-infected mouse lungs enables imaging of mycobacterial granulomas deep within tissue to a depth of more than 1 mm. Using established transgenic lines, we were able to image normal and pathogenic structures and their surrounding host context at high resolution. We identified the three-dimensional organization of granuloma-associated angiogenesis, an important feature of mycobacterial infection, and characterized the induction of the cytokine tumor necrosis factor (TNF) within the granuloma using an established fluorescent reporter line. We observed heterogeneity in TNF induction within granuloma macrophages, consistent with an evolving view of the tuberculous granuloma as a non-uniform, heterogeneous structure. Broad application of this technique will enable new understanding of host-pathogen interactions in situ.

Synthesis and evaluation of novel functionalised indoles as anticancer agents

This thesis outlines the design and application of new routes towards a range of novel bisindolylmaleimide and indolo[2,3-a]carbazole derivatives, and evaluation of their biological effects and their chemotherapeutic potential. A key part of this work focussed on utilising a hydroxymaleimide as a replacement for the prevalent lactam/maleimide functionality and forming a series of novel derivatives through substitution on the indole nitrogens. To achieve this, a robust synthetic strategy was developed which allowed access to key maleic anhydride intermediates using Perkin-type methodology. These hydroxymaleimides were further modified via a Lossen rearrangement to furnish a series of analogues containing a 6-membered F-ring. The theme of F-ring modulation was further expanded through the utilisation of a second route involving the design and synthesis of β-keto ester intermediates, which afforded novel derivatives containing pyrazolone and isocytosine headgroups, and various N-substituents. Work on a further route involving a dione intermediate resulted in the isolation of a bisindolyl derivative with a novel imidazole F-ring. Following the synthesis of 42 novel compounds, extensive screening was undertaken using the NCI-60 cell line screen, with twelve candidates progressing to evaluation via the five dose assay. This led to the identification of several lead compounds with high cytotoxicity and excellent selectivity profiles, which included derivatives with low nanomolar GI50 values against specific cancer cell lines, and also derivatives with selective cytotoxicity. Preliminary results from a kinase screen indicated noteworthy selectivity towards GSK3α/β and PIM1 kinases, with low micromolar IC50 values being observed for these enzymes.

Organic geochemistry of sediments from sapropel 94 of ODP Hole 161-974C

Analysis of the molecular composition of the organic matter (OM) from whole sediment samples can avoid analytical bias that might result from isolation of components from the sediment matrix, but has its own analytical challenges. We evaluated the use of GC * GC-ToFMS to analyze the pyrolysis products of six whole sediment samples obtained from above, within and below a 1 million year old OM-rich Mediterranean sapropel layer. We found differences in pyrolysis products of alkyl pyrroles, probably derived from chlorophyll, in pyrolysates of the sapropels but not in those of the marls suggests that higher marine productivity and greater OM preservation accompanied deposition of the sapropels. Detection of tetramethyl benzenes considered to be pyrolysis products of isorenieratene in the sapropel samples is evidence that nitrogen-fixing green sulfur bacteria contributed to the high productivity. Greater abundances of shorter chain aliphatic hydrocarbons, pyrroles, furans and alkyl aromatics in the pyrolysates of sapropel samples relative to the marls confirm better preservation of marine OM in the sapropels. In addition, the presence of greater amounts of thiophenes in the sapropels than in the marls is consistent with the existence of euxinic conditions during sapropel deposition. The combination of whole sediment pyrolysis and GC × GC-ToFMS is promising, but the procedure requires careful selection of its multiple analytical variables, particularly the pyrolysis temperature and the operational features of the GC columns.