Norethisterone oenanthate as an injectable contraceptive: use of a modified dose schedule.

Norethisterone oenanthate (NET-OEN) was given as an injectable contraceptive to 295 healthy women over 1606 woman-months. A modified injection schedule was used. There were no pregnancies, and the 12-month, life-table, use-related discontinuation rate was 39.1/100 users. Menstrual disturbance (10.8/100 women), minor side effects (13.5/100 women), and personal reasons (12.0/100 women) were the main causes of use-related discontinuation. There was no difference in use-related discontinuation rates between women receiving their first injection during a normal menstrual period and those receiving it immediately after a pregnancy. There were no serious side effects. The use of NET-OEN in certain groups of women is recommended, particularly in those in need of highly effective contraception, who cannot or do not wish to take oral contraceptives, who are lactating, or who are awaiting hospital admission for sterilisation.     

REDUCTION OF THE PECTORAL SPINE AND GIRDLE IN DOMESTICATED CHANNEL CATFISH IS LIKELY CAUSED BY CHANGES IN SELECTION PRESSURE

Locked pectoral spines of the Channel Catfish Ictalurus punctatus more than double the fish's width and complicate ingestion by gape‐limited predators. The spine mates with the pectoral girdle, a robust structure that anchors the spine. This study demonstrates that both spine and girdle exhibit negative allometric growth and that pectoral spines and girdles are lighter in domesticated than in wild Channel Catfish. This finding could be explained by changes in selection pressure for spine growth during domestication or by an epigenetic effect in which exposure to predators in wild fish stimulates pectoral growth. We tested the epigenetic hypothesis by exposing domesticated Channel Catfish fingerlings to Largemouth Bass Micropterus salmoides predators for 13 weeks. Spines and girdles grow isometrically in the fingerlings, and regression analysis indicates no difference in proportional pectoral growth between control and predator‐exposed fish. Therefore a change in selection pressure likely accounts for smaller pectoral growth in domesticated Channel Catfish. Decreasing spine growth in older fish suggests anti‐predator functions are most important in smaller fish. Additionally, growth of the appendicular and axial skeleton is controlled differentially, and mechanical properties of the spine and not just its length are an important component of this defensive adaptation.     

Pharmacological characterization of INCB3344, a small molecule antagonist of human CCR2

The chemokine receptor 2 (CCR2) directs migration of monocytes and has been proposed to be a drug target for chronic inflammatory diseases. INCB3344 was first published as a small molecule nanomolar inhibitor of rodent CCR2. Here, we show that INCB3344 can also bind human CCR2 (hCCR2) with high affinity, having a dissociation constant (K_d) of approximately 5 nM. The binding of the compound to the receptor is rapid and reversible. INCB3344 potently inhibits hCCR2 binding of monocyte chemoattractant protein-1 (MCP-1) and MCP-1-induced signaling and function in hCCR2-expressing cells, including ERK phosphorylation and chemotaxis, and is competitive against MCP-1 in vitro. INCB3344 also blocks MCP-1 binding to monocytes in human whole blood, with potency consistent with in vitro studies. The whole blood binding assay described here can be used for monitoring pharmacodynamic activity of CCR2 antagonists in both preclinical models and in the clinic.     

Analyses of Current-Generating Mechanisms of Shewanella loihica PV-4 and Shewanella oneidensis MR-1 in Microbial Fuel Cells

Although members of the genus Shewanella have common features (e.g., the presence of decaheme c-type cytochromes [c-cyts]), they are widely variable in genetic and physiological features. The present study compared the current-generating ability of S. loihica PV-4 in microbial fuel cells (MFCs) with that of well-characterized S. oneidensis MR-1 and examined the roles of c-cyts in extracellular electron transfer. We found that strains PV-4 and MR-1 exhibited notable differences in current-generating mechanisms. While the MR-1 MFCs maintained a constant current density over time, the PV-4 MFCs continued to increase in current density and finally surpassed the MR-1 MFCs. Coulombic efficiencies reached 26% in the PV-4 MFC but 16% in the MR-1 MFCs. Although both organisms produced quinone-like compounds, anode exchange experiments showed that anode-attached cells of PV-4 produced sevenfold more current than planktonic cells in the same chamber, while planktonic cells of MR-1 produced twice the current of the anode-attached cells. Examination of the genome sequence indicated that PV-4 has more c-cyt genes in the metal reductase-containing locus than MR-1. Mutational analysis revealed that PV-4 relied predominantly on a homologue of the decaheme c-cyt MtrC in MR-1 for current generation, even though it also possesses two homologues of the decaheme c-cyt OmcA in MR-1. These results suggest that current generation in a PV-4 MFC is in large part accomplished by anode-attached cells, in which the MtrC homologue constitutes the main path of electrons toward the anode.Some species of dissimilatory metal-reducing bacteria (DMRB) are able to reduce solid metal oxides as terminal electron acceptors and generate currents in microbial fuel cells (MFCs) (2, 11, 14, 30, 46). Although mixed cultures are often used in MFC experiments (13), studies seeking a mechanistic understanding of electron transfer to electrode surfaces typically target pure cultures of such DMRB, due to the complexity in microbial communities. Presently, two model DMRB, Shewanella oneidensis MR-1 and Geobacter sulfurreducens PCA (2, 3, 12, 18, 31), are used in most investigations.S. oneidensis MR-1 is a metabolically diverse DMRB that has been studied extensively for its potential use in bioremediation applications. For this reason, MR-1 was the first Shewanella species to have its genome completely sequenced and annotated (10). In addition, since the first report in 1999 when this microorganism was shown to have the ability to transfer electrons to the electrode without an exogenously added mediator (14), it has also become one of the model organisms for the study of electron transfer mechanisms in MFCs.Although the molecular mechanisms for extracellular electron transfer have not yet been elucidated fully, c-type cytochromes (c-cyts) appear to be the key cellular components involved in this process (38). In S. oneidensis MR-1, OmcA and MtrC are outer membrane (OM), decaheme c-cyts that are considered to be involved in the direct (directly attached) electron transfer to solid metal oxides and anodes of MFCs (9, 20, 22, 23, 47). Several pieces of evidence suggest that OmcA and MtrC form a complex and act in a cooperative manner (33, 37, 42), and these results correlate with the fact that the genes encoding these proteins constitute an operon-like cluster in the chromosome (1). It has also been shown that MtrC and OmcA have overlapping functions as terminal reductases of metal oxides (25, 38). OmcA and MtrC are also present on the surface of nanowires and may be involved in the long-range transfer of electrons (8). In addition to direct electron transfer, MR-1 has the ability to produce water-soluble electron-shuttle compounds (quinones and flavins) that are involved in the mediated electron transfer from cells to distant solid electron acceptors (metal oxides or MFC anodes) (21, 27, 44).Recently, the genome sequences of nearly 20 Shewanella strains have been completed and annotated, opening the door to study the diversity of their extracellular electron transfer mechanisms. A comparison of their genomes has shown that although they have some consensus OM c-cyt genes, variations exist in the number and order of these genes in their metal reductase-containing loci (6). One such species is S. loihica strain PV-4, which was recently isolated from an iron-rich microbial mat near a deep-sea hydrothermal vent located on the Loihi Seamount in Hawaii (7, 32). The phenotypic and phylogenetic characteristics of PV-4 were determined, with a subsequent study focusing on the metal reduction and iron biomineralization capabilities of this bacterium (32). Initial experiments performed in our laboratory revealed that PV-4 developed a c-cyt-dependent deep red color that was much more striking than that of strain MR-1 when grown anaerobically with iron oxide as the terminal electron acceptor (26). This allowed us to assume that PV-4 could have a high extracellular electron transfer ability. Accordingly, the present study evaluated the current-producing ability of strain PV-4 in MFCs and examined the roles of some c-cyts in extracellular electron transfer. Special attention was paid to the comparison of PV-4 with MR-1 to reveal differences in mechanisms for extracellular electron transfer. We report herein differences between these strains in the roles of OM c-cyts for extracellular electron transfer, the behaviors and metabolic patterns of MFC, and the resultant MFC performances.     

Glucosinolate polymorphism in wild cabbage (Brassica oleracea) influences the structure of herbivore communities

Natural plant populations often show substantial heritable variation in chemical structure of secondary metabolites. Despite a great deal of evidence from laboratory studies that these chemicals influence herbivore behaviour and life history, there exists little evidence for the structuring of natural herbivore communities according to plant chemical profiles. Brassica oleracea (Brassicaceae) produces aliphatic glucosinolates, which break down into toxins when leaf tissue is damaged. Structural diversity in these glucosinolates is heritable, and varies considerably at two ecological scales in the UK: both within and between populations. We surveyed herbivore attack on plants producing different glucosinolates, using 12 natural B. oleracea populations. In contrast to the results of previous studies in this system, which suffered low statistical power, we found significant differential responses of herbivore species to heritable glucosinolates, both within and between plant populations. We found significant correlations between herbivore infestation rates and the presence or absence of two heritable glucosinolates: sinigrin and progoitrin. There was variation between herbivore species in the direction of response, the ecological scale at which responses were identified, and the correlations for some herbivore species changed at different times of the year. We conclude that variation in plant secondary metabolites can structure the community of herbivores that attack them, and propose that herbivore-mediated differential selection deserves further investigation as a mechanism maintaining the observed diversity of glucosinolates in wild Brassica.