The role of polyamine architecture on the pharmacological activity of open lactone camptothecin-polyamine conjugates

A series of camptothecin open-ring lactone tripartate conjugates were synthesized, in which polyamine side chains with different architecture (ethane-1,2-diamine, spermidine, homospermidine, spermine, and 4,8,13,17-tetraza-icosane-1,20-diamine) are linked to the 21-carboxylic function through an amidic bond, while the 17-CH(2)OH is acetylated. The rationale for the synthesis of these compounds was to explore the influence of the polyamine architecture on the activity of these CPT conjugates into cells, since the positively charged ammonium cations would favor interaction through electrostatic binding to the negatively charged DNA backbone. Topoisomerase I-mediated DNA cleavage assay was used to investigate the ability of these compounds to stimulate the DNA damage. The cleavage pattern was found to be similar to that of SN38 for all the new CPTs. The CPT tripartates were tested for growth inhibition ability against the human non-small-cell lung cancer carcinoma NCI-H460 cell line. Although these compounds were less potent than topotecan, SN38, and CPT after 1 h of treatment, the antiproliferative effects greatly increased after 72 h of exposure. The growth inhibition potency during long-term exposure is correlated with the number of charges of the 21-amide substituent. Both cleavage assay and in vitro effects support the interpretation that the compounds may have inhibitory activity also in the open-ring form. The architecture of the polyamine moiety is important for antiproliferative activity, and a balance between the hydrophilic and lipophilic properties of the polyamine is critical for CPT potency.     

The use and limits of AFLP data in the taxonomy of polyploid wild potato species in Solanum series Conicibaccata

Solanum sect. Petota (tuber-bearing wild and cultivated potatoes) are a group of approximately 190 wild species distributed throughout the Americas from the southwestern United States south to Argentina, Chile, and Uruguay. Solanum series Conicibaccata are a group of approximately 40 species within sect. Petota, distributed from central Mexico to central Bolivia, composed of diploids (2n = 2x = 24), tetraploids (2n = 4x = 48) and hexaploids (2n = 6x = 64); the polyploids are thought to be polysomic polyploids. This study initially was designed to address species boundaries of the four Mexican and Central American species of series Conicibaccata with AFLP data with the addition of first germplasm collections of one of these four species, Solanum woodsonii, as a follow-up to prior morphological, chloroplast DNA, and RAPD studies; and additional species of series Conicibaccata from South America. AFLP data from 12 primer combinations (1722 polymorphic bands) are unable to distinguish polyploid species long thought to be distinct. The data suggest a complex reticulate history of the tetraploids or the need for a broad downward reevaluation of the number of species in series Conicibaccata, a trend seen in other series of sect. Petota. Separately, through flow cytometry, we report the first ploidy level of S. woodsonii, as tetraploid (2n = 48). The U.S. Government’s right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

Investigations in commonness and rarity: a comparative analysis of co-occurring, congeneric Mexican trees

Population size distributions were examined for 12 species of trees co-occurring at Chamela Biological Station in Jalisco, Mexico. Species had been selected as congeneric pairs and trios similar in gross morphology and ecology in order better to identify correlates of relative abundance. Rarer species were found unanimously to have more irregular distributions of individuals among size classes than more common species when distributions were compared to a smooth, descending curve constructed from population mean stem diameters (an exponential distribution). Examination among species of patterns of deviation from these corresponding smooth distributions indicates that the most reasonably inferred cause for the observed pattern is consistent differences in degree of fluctuation in recruitment into adult size classes. These results thereby suggest a demographic difference between locally rarer and more common species that may be generally associated with observed differences in relative abundance and indicate a focus for management of rarity in forest trees.     

Feather mites on birds: costs of parasitism or conditional outcomes?

Feather mites (suborder Astigmata, superfamilies Analgoidea, Pterolichoidea and Freyaniidae) are among the commonest ectosymbionts of birds. Most researchers have assumed they are parasites, having negative effects on hosts. Here we present evidence that suggests that feather mites may not be parasites. We develop a framework for considering conditional outcomes in these interspecific associations, dealing with different kinds of relationships between symbionts. The non-parasitic status of feather mites is supported by a literature review as well as by preliminary data on mites' food. We illustrate symbiotic relationships with a graphical model showing different scenarios in which hosts' cost-benefit relations are determined by the interactions among their symbionts.     

Photocarotenogenesis in Phycomyces: Expression of the carB Gene Encoding Phytoene Dehydrogenase

Phycomyces blakesleeanus the biosynthesis of β-carotene is under the control of blue light. The light-controlled expression of the carB gene encoding phytoene dehydrogenase was investigated with slot blot and Northern analyses. After irradiation of mycelia with short pulses of blue light the amount of carB mRNA was stimulated transiently in the subsequent dark period. Depending on the energy fluence of the light pulse the mRNA increased about 3 to 5-fold after a latency of 2-5 min. Twenty minutes after the irradiation the amount of carB mRNA decreased again and reached, after 60 min, the dark level it had prior to the irradiation. A Northern analysis showed that the carB mRNA is unstable. After irradiation the full-length mRNA (2.2 kb) as well as degraded carB mRNA increased. The degradation was specific for the carB gene, because no degradation was observed for 25S rRNA or the mRNA of the pyrG gene. The rapid turnover as well as the degradation of the carB mRNA provide for an adaptive light control and appear to represent an essential feature of the regulation of the carotene pathway. Received 11 April 2000/ Accepted in revised form 30 November 2000     

Pollen availability and abundance of predatory phytoseiid mites on natural and secondary hedgerows

Mites occurring on a number ofnatural and secondary (i.e., planted) hedgerowslocated in north-eastern Italy were monitoredwith particular attention being paid tophytoseiids (Acari: Phytoseiidae). Pollendiversity and abundance were analysed tohighlight potential relationships betweenpollen availability and phytoseiids. Secondaryhedgerows were structured in modules containinga single species (field maple, hornbeam,dogwood, hazel, and elderberry) andmodules containing a mixture of theabove-mentioned species. These hedgerows werelocated in an experimental farm where they werecontiguous to a vineyard. Natural hedgerowswere located in the same area. The pollen ofPoaceae was the most important component ofnon-arboreal pollen (NAP), and that ofelderberry of arboreal pollen (AP). Pollendiversity was higher on natural than onsecondary hedgerows. Generally, pollendensities reached relatively high levels fromlate April to late June and decreased insummer. Pollen was more abundant on naturalthan on secondary hedgerows and phytoseiidabundance showed a similar trend. On elderberryand hornbeam in natural hedgerows a potentialrelationship between pollen availability andthe abundance of phytoseiids, in particular ofEuseius finlandicus (Oudemans) was found.Experimental pollen applications increased thefecundity and the abundance of this species.     

A Sox10 expression screen identifies an amino acid essential for Erbb3 function

The neural crest (NC) is a population of embryonic stem cells that gives rise to numerous cell types, including the glia and neurons of the peripheral and enteric nervous systems and the melanocytes of the skin and hair. Mutations in genes and genetic pathways regulating NC development lead to a wide spectrum of human developmental disorders collectively called neurocristopathies. To identify molecular pathways regulating NC development and to understand how alterations in these processes lead to disease, we established an N-ethyl-N-nitrosourea (ENU) mutagenesis screen utilizing a mouse model sensitized for NC defects, Sox10^LacZ/+. Out of 71 pedigrees analyzed, we identified and mapped four heritable loci, called modifier of Sox10 expression pattern 1–4 (msp1–4), which show altered NC patterning. In homozygous msp1 embryos, Sox10^LacZ expression is absent in cranial ganglia, cranial nerves, and the sympathetic chain; however, the development of other Sox10-expressing cells appears unaffected by the mutation. Linkage analysis, sequencing, and complementation testing confirmed that msp1 is a new allele of the receptor tyrosine kinase Erbb3, Erbb3^msp1, that carries a single amino acid substitution in the extracellular region of the protein. The ENU-induced mutation does not alter protein expression, however, it is sufficient to impair ERBB3 signaling such that the embryonic defects observed in msp1 resemble those of Erbb3 null alleles. Biochemical analysis of the mutant protein showed that ERBB3 is expressed on the cell surface, but its ligand-induced phosphorylation is dramatically reduced by the msp1 mutation. These findings highlight the importance of the mutated residue for ERBB3 receptor function and activation. This study underscores the utility of using an ENU mutagenesis to identify genetic pathways regulating NC development and to dissect the roles of discrete protein domains, both of which contribute to a better understanding of gene function in a cellular and developmental setting.     

Antibody action after phagocytosis promotes Cryptococcus neoformans and Cryptococcus gattii macrophage exocytosis with biofilm-like microcolony formation

Antibody-mediated phagocytosis was discovered over a century ago but little is known about antibody effects in phagolysosomes. We explored the consequences of antibody-mediated phagocytosis for two closely related human pathogenic fungal species, Cryptococcus neoformans and Cryptococcus gattii , of which C. neoformans encompasses two varieties: neoformans and grubii. The interaction between C. neoformans varieties grubii and neoformans and host cells has been extensively studied, but that of C. gattii and macrophages remains largely unexplored. Like C. neoformans , antibody-mediated phagocytosis of C. gattii cells was followed by intracellular replication, host cell cytoplasmic polysaccharide accumulation and phagosomal extrusion. Both C. gattii and C. neoformans cells exited macrophages in biofilm-like microcolonies where the yeast cells were aggregated in a polysaccharide matrix that contained bound antibody. In contrast, complement-opsonized C. neoformans variety grubii cells were released from macrophages dispersed as individual cells. Hence, both antibody- and complement-mediated phagocytosis resulted in intracellular replication but the mode of opsonization affected the outcome of exocytosis. The biofilm-like microcolony exit strategy of C. neoformans and C. gattii following antibody opsonization reduced fungal cell dispersion. This finding suggests that antibody agglutination effects persist in the phagosome to entangle nascent daughter cells and this phenomenon may contribute to antibody-mediated protection.     

Passive administration of monoclonal antibodies to Anthrolysin O prolong survival in mice lethally infected with <Emphasis Type="Italic">Bacillus anthracis</Emphasis>

Background  

Bacillus anthracis has two major virulence factors: a tripartite toxin that produces lethal and edema toxins and a polyglutamic acid capsule. A recent report suggested that a toxin belonging to the cholesterol dependant cytolysin (CDC) family, anthrolysin O (ALO) was a new virulence factor for B. anthracis but subsequent studies have questioned its relevance in pathogenesis. In this study, we examined the immunogenicity of recombinant anthrolysin O (rALO) in mice.     

Rapid Development and Optimization of Tablet Manufacturing Using Statistical Tools

The purpose of this paper was to develop a statistical methodology to optimize tablet manufacturing considering drug chemical and physical properties applying a crossed experimental design. The assessed model drug was dried ferrous sulphate and the variables were the hardness and the relative proportions of three excipients, binder, filler and disintegrant. Granule properties were modeled as a function of excipient proportions and tablet parameters were defined by the excipient proportion and hardness. The desirability function was applied to achieve optimal values for excipient proportions and hardness. In conclusion, crossed experimental design using hardness as the only process variable is an efficient strategy to quickly determine the optimal design process for tablet manufacturing. This method can be applied for any tablet manufacturing method.