Surface Expression and Subunit Specific Control of Steady Protein Levels by the Kv7.2 Helix A-B Linker

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Workshop on climate change and salmon production, Vancouver, March 26-27, 1998: technical report

The Workshop on Climate Change and Salmon Production was held in Vancouver, Canada, 26-27 March 1998. The Workshop was organized and sponsored by the North Pacific Anadromous Fish Commission (NPAFC). Each Party to the Commission designated one scientist to the Workshop Steering Committee. Each member of the Steering Committee chaired one half-day session of the Workshop. All necessary arrangements were made by the NPAFC Secretariat in cooperation with the Steering Committee and the Canadian Party to the Commission. (PDF contains 60 pages) Over 70 scientists, industry representatives and fisheries officials attended the Workshop. There were 20 presentations of scientific papers followed by the discussion sessions. Extended abstracts are included in this Technical Report, which also contains opening address by the Chairman of the Steering Committee and short review of the Workshop by the Coordinator. The material presented in the Technical Report has not been peer reviewed and does not necessarily reflect the views of either the NPAFC or the Parties. The material has been edited by the technical editor for clarity and publication purposes only. Items in this Report should not be cited except as personal communication and with the author's permission.

Detection of hepatitis B virus markers using a biosensor based on imaging ellipsometry

A biosensor based on imaging ellipsometry (BIE) has been developed and validated in 169 patients for detecting five markers of hepatitis B virus (HBV) infection. The methodology has been established to pave the way for clinical diagnosis, including ligand screening, determination of the sensitivity, set-up of cut-off values (CoVs) and comparison with other clinical methods. A matrix assay method was established for ligand screening. The CoVs of HBV markers were derived with the help of receiver operating characteristic curves. Enzyme-linked immunosorbent assay (ELISA) was the reference method. Ligands with high bioactivity were selected and sensitivities of 1 ng/mL and 1 IU/mL for hepatitis B surface antigen (HBsAg) and surface antibody (anti-HBs) were obtained respectively. The CoVs of HBsAg, anti-HBs, hepatitis B e antigen, hepatitis B e antibody and core antibody were as follows: 15%, 18%, 15%, 20% and 15%, respectively, which were the percentages over the values of corresponding ligand controls. BIE can simultaneously detect up to five markers within 1 h with results in acceptable agreement with ELISA, and thus shows a potential for diagnosing hepatitis B with high throughput.

Report on estimating the size of dolphin schools, based on data obtained during a charter cruise of the M/V Gina Anne, October 11 -November 25, 1979

Estimates of dolphin school sizes made by observers and crew members aboard tuna seiners or by observers on ship or aerial surveys are important components of population estimates of dolphins which are involved in the yellowfin tuna fishery in the eastern Pacific. Differences in past estimates made from tuna seiners and research ships and aircraft have been noted by Brazier (1978). To compare various methods of estimating dolphin school sizes a research cruise was undertaken with the following major objectives: 1) compare estimates made by observers aboard a tuna seiner and in the ship's helicopter, from aerial photographs, and from counts made at the backdown channel, 2) compare estimates of observers who are told the count of the school size after making their estimate to the observer who is not aware of the count to determine if observers can learn to estimate more accurately, and 3) obtain movie and still photographs of dolphin schools of known size at various stages of chase, capture and release to be used for observer training. The secondary objectives of the cruise were to: 1) obtain life history specimens and data from any dolphins that were killed incidental to purse seining. These specimens and data were to be analyzed by the U.S. National Marine Fisheries Service ( NMFS ) , 2) record evasion tactics of dolphin schools by observing them from the helicopter while the seiner approached the school, 3) examine alternative methods for estimating the distance and bearing of schools where they were first sighted, 4) collect the Commission's standard cetacean sighting, set log and daily activity data and expendable bathythermograph data. (PDF contains 31 pages.)

Comparative growth and survival of fry of Clarias gariepinus (B), Clarias anguillaris (L), Heterobranchus bidorsalis (G), Heteroclarias, Clariabranchus and Clarias hybrids under outdoor nursery management system

Hybrids of Clariid catfishes; C. gariepinus (Netherlands), C. anguillaris, H. bidorsalis and their parental species were monitored for 8 weeks in 2 x 2 x 1m outdoor concrete tanks. The fry were fed NIFFR diet (40% crude protein) twice daily, 7 days of the week. Growth and survival records were taken weekly. The male HEB X female CLG hybrid showed an overall highest performance in growth rate while the lowest was recorded in male CLA X female CLG hybrid. The male HEB X female CLG hybrid grew at a faster rate than its reciprocal hybrid. In view or their growth rate, it is possible that the growth and survival rates or H. bidorsalis especially at the fry to fingerling stage could be improved through hybridization. The hybrid have potential as commercial food fish

Alaves B taldearen lesioak 2012-2013 denboraldian: azterketa eta hobetzeko ardatzak

Futbolean iskiotibialetan ematen diren lesioen errebisioa eta hauek ekiditeko proposamena.

Function of microRNA-146a and NF-κB in physiologic and pathologic hematopoiesis

During inflammation and infection, hematopoietic stem and progenitor cells (HSPCs) are stimulated to proliferate and differentiate into mature immune cells, especially of the myeloid lineage. MicroRNA-146a (miR-146a) is a critical negative regulator of inflammation. Deletion of the gene encoding miR-146a—expressed in all blood cell types—produces effects that appear as dysregulated inflammatory hematopoiesis, leading to a decline in the number and quality of hematopoietic stem cells (HSCs), excessive myeloproliferation, and, ultimately, to exhaustion of the HSCs and hematopoietic neoplasms. Six-week-old deleted mice are normal, with no effect on cell numbers, but by 4 months bone marrow hypercellularity can be seen, and by 8 months marrow exhaustion is becoming evident. The ability of HSCs to replenish the entire hematopoietic repertoire in a myelo-ablated mouse also declines precipitously as miR-146a-deficient mice age. In the absence of miR-146a, LPS-mediated serial inflammatory stimulation accelerates the effects of aging. This chronic inflammatory stress on HSCs in deleted mice involves a molecular axis consisting of upregulation of the signaling protein TRAF6 leading to excessive activity of the transcription factor NF-κB and overproduction of the cytokine IL-6. At the cellular level, transplant studies show that the defects are attributable to both an intrinsic problem in the miR-146a-deficient HSCs and extrinsic effects of miR-146a-deficient lymphocytes and non-hematopoietic cells. This study has identified a microRNA, miR-146a, to be a critical regulator of HSC homeostasis during chronic inflammatory challenge in mice and has provided a molecular connection between chronic inflammation and the development of bone marrow failure and myeloproliferative neoplasms. This may have implications for human hematopoietic malignancies, such as myelodysplastic syndrome, which frequently displays downregulated miR-146a expression.

Measurement of the direct CP asymmetry in b-->s+gamma via sum of exclusive B Meson decays using the BABAR detector

We perform a measurement of direct CP violation in b to s+gamma Acp, and the measurement of a difference between Acp for neutral B and charged B mesons, Delta A_{X_s\gamma}, using 429 inverse femtobarn of data recorded at the Upsilon(4S) resonance with the BABAR detector. B mesons are reconstructed from 16 exclusive final states. Particle identification is done using an algorithm based on Error Correcting Output Code with an exhaustive matrix. Background rejection and best candidate selection are done using two decision tree-based classifiers. We found $\acp = 1.73%+-1.93%+-1.02% and Delta A_X_sgamma = 4.97%+-3.90%+-1.45% where the uncertainties are statistical and systematic respectively. Based on the measured value of Delta A_X_sgamma, we determine a 90% confidence interval for Im C_8g/C_7gamma, where C_7gamma and C_8g are Wilson coefficients for New Physics amplitudes, at -1.64 < Im C_8g/C_7gamma < 6.52.

Protecting a threatened coastal fish species through regional collaboration

Rainbow smelt (Osmerus mordax) are small anadromous fish that live in nearshore coastal waters during much of the year and migrate to tidal rivers to spawn during the spring. They are a key prey species in marine food webs, as they are consumed by larger organisms such as striped bass, bluefish, and seabirds. In addition, smelt are valued culturally and economically, as they support important recreational and commercial fisheries. The Atlantic Coast range of rainbow smelt has been contracting in recent decades. Historically, populations extended from the Delaware River to eastern Labrador and the Gulf of St. Lawrence (Buckley 1989). More recent observations indicate that rainbow smelt spawning populations have been extirpated south of Long Island Sound, and evidence of spawning activity is extremely limited between Long Island and Cape Cod, MA. In the Gulf of Maine region, spawning runs are still observed, but monitoring surveys as well as commercial and recreational catches indicate that these populations have also declined (e.g., Chase and Childs 2001). Many diverse factors could drive the recently noted declines in rainbow smelt populations, including spawning habitat conditions, fish health, marine environmental conditions, and fishing pressure. Few studies have assessed any of these potential threats or their joint implications. In 2004, the National Marine Fisheries Service (NMFS) listed rainbow smelt as a species of concern. Subsequently, the states of Maine, New Hampshire, and Massachusetts were awarded a grant through NMFS’s Proactive Conservation Program to gather new information on the status of rainbow smelt, identify factors that affect spawning populations, and develop a multi-state conservation program. This paper provides an overview of this collaborative project, highlighting key biological monitoring and threats assessment research that is being conducted throughout the Gulf of Maine. (PDF contains 4 pages)

The intramolecular Buchner reaction of α-diazo-β-ketonitriles : development and application to the total synthesis of (+)-Salvileucalin B

<p>Since its discovery in 1896, the Buchner reaction has fascinated chemists for more than a century. The highly reactive nature of the carbene intermediates allows for facile dearomatization of stable aromatic rings, and provides access to a diverse array of cyclopropane and seven-membered ring architectures. The power inherent in this transformation has been exploited in the context of a natural product total synthesis and methodology studies.</p> <p>The total synthesis work details efforts employed in the enantioselective total synthesis of (+)-salvileucalin B. The fully-substituted cyclopropane within the core of the molecule arises from an unprecedented intramolecular Buchner reaction involving a highly functionalized arene and an α-diazo-β-ketonitrile. An unusual retro-Claisen rearrangement of a complex late-stage intermediate was discovered on route to the natural product.</p> <p>The unique reactivity of α-diazo-β-ketonitriles toward arene cyclopropanation was then investigated in a broader methodological study. This specific di-substituted diazo moiety possesses hitherto unreported selectivity in intramolecular Buchner reactions. This technology was enables the preparation of highly functionalized norcaradienes and cyclopropanes, which themselves undergo various ring opening transformations to afford complex polycyclic structures.</p> <p>Finally, an enantioselective variant of the intramolecular Buchner reaction is described. Various chiral copper and dirhodium catalysts afforded moderate stereoinduction in the cyclization event.</p>

Sequence specific complexation of b DNA at sites containing g,c base pairs

<p>A series of eight related analogs of distamycin A has been synthesized. Footprinting and affinity cleaving reveal that only two of the analogs, pyridine-2- car box amide-netropsin (2-Py N) and 1-methylimidazole-2-carboxamide-netrops in (2-ImN), bind to DNA with a specificity different from that of the parent compound. A new class of sites, represented by a TGACT sequence, is a strong site for 2-PyN binding, and the major recognition site for 2-ImN on DNA. Both compounds recognize the G•C bp specifically, although A's and T's in the site may be interchanged without penalty. Additional A•T bp outside the binding site increase the binding affinity. The compounds bind in the minor groove of the DNA sequence, but protect both grooves from dimethylsulfate. The binding evidence suggests that 2-PyN or 2-ImN binding induces a DNA conformational change.</p> <p>In order to understand this sequence specific complexation better, the Ackers quantitative footprinting method for measuring individual site affinity constants has been extended to small molecules. MPE•Fe(II) cleavage reactions over a 10^5 range of free ligand concentrations are analyzed by gel electrophoresis. The decrease in cleavage is calculated by densitometry of a gel autoradiogram. The apparent fraction of DNA bound is then calculated from the amount of cleavage protection. The data is fitted to a theoretical curve using non-linear least squares techniques. Affinity constants at four individual sites are determined simultaneously. The distamycin A analog binds solely at A•T rich sites. Affinities range from 10^(6)- 10^(7)M^(-1) The data for parent compound D fit closely to a monomeric binding curve. 2-PyN binds both A•T sites and the TGTCA site with an apparent affinity constant of 10^(5) M^(-1). 2-ImN binds A•T sites with affinities less than 5 x 10^(4) M^(-1). The affinity of 2-ImN for the TGTCA site does not change significantly from the 2-PyN value. At the TGTCA site, the experimental data fit a dimeric binding curve better than a monomeric curve. Both 2-PyN and 2-ImN have substantially lower DNA affinities than closely related compounds.</p> <p>In order to probe the requirements of this new binding site, fourteen other derivatives have been synthesized and tested. All compounds that recognize the TGTCA site have a heterocyclic aromatic nitrogen ortho to the N or C-terminal amide of the netropsin subunit. Specificity is strongly affected by the overall length of the small molecule. Only compounds that consist of at least three aromatic rings linked by amides exhibit TGTCA site binding. Specificity is only weakly altered by substitution on the pyridine ring, which correlates best with steric factors. A model is proposed for TGTCA site binding that has as its key feature hydrogen bonding to both G's by the small molecule. The specificity is determined by the sequence dependence of the distance between G's.</p> <p>One derivative of 2-PyN exhibits pH dependent sequence specificity. At low pH, 4-dimethylaminopyridine-2-carboxamide-netropsin binds tightly to A•T sites. At high pH, 4-Me_(2)NPyN binds most tightly to the TGTCA site. In aqueous solution, this compound protonates at the pyridine nitrogen at pH 6. Thus presence of the protonated form correlates with A•T specificity.</p> <p>The binding site of a class of eukaryotic transcriptional activators typified by yeast protein GCN4 and the mammalian oncogene Jun contains a strong 2-ImN binding site. Specificity requirements for the protein and small molecule are similar. GCN4 and 2-lmN bind simultaneously to the same binding site. GCN4 alters the cleavage pattern of 2-ImN-EDTA derivative at only one of its binding sites. The details of the interaction suggest that GCN4 alters the conformation of an AAAAAAA sequence adjacent to its binding site. The presence of a yeast counterpart to Jun partially blocks 2-lmN binding. The differences do not appear to be caused by direct interactions between 2-lmN and the proteins, but by induced conformational changes in the DNA protein complex. It is likely that the observed differences in complexation are involved in the varying sequence specificity of these proteins.</p>

Experimente über die Wirkung von UV-B auf pelagische Fischembryonen in einem Sonnenscheinsimulator

One of the supposed effects of the observed ozone depletion is the increase of solar UV-B irradiation at the seasurface. This will cause an impact on certain compartments of marine ecosystems. Especially, sensitive developmental stages of pelagic fish embryos might be affected. Embryos of dab (Limanda limanda) and plaice (Pleuronectes plalessa) were experimentally exposed 10 different amounts of UVB irradiation in a sunshine simulator. This programmable device allows the dosage of realistic solar irradiation in quality and guantity. Experiments were carried out in March 1995 and February 1996. Either artificially inserninated and reared emhryos of dab and plaice or embryos caught in the German Bight were exposed to simulated solar irradiation. The 1995 experiments served to identify the effective irradiation dosages. For the 1996 experiments irradiation applied was much lower, being dose to realistic valucs expected over the North Sea as a consequence of ozone depletion. The following end points were studied: 1. Mortality, 2. sublethal morphological effects (malformations), 3. DNA damage, 4. changes in buoyancy of embryos measured as changes in osmolarity of the perivitelline fluid. Conditions for the simulation of daylight were a c1oudless sky with a solar zenith distance of 34 % (air mass 1.2). The adopted ozone depletion was 40 % corresponding to 180 DU (Dobson Units) instead of 300 DU. In the 1995 experiments time and dosage dependent influenccs on mortality and buoyancy of embryos of dab and plaice were found. Even in those embryos which were protected from the UV-B spectral range a loss of buoyancy was registered after 12 hours in the simulator. No diffcrences in DNA integrity as determined by DNA unwinding of exposed and control embryos were found. Also with lower amounts of irradiation in the 1996 experiments dosage dependent acute mortality, malformations, and impact on the buoyancy of the emhryos was registered. Sublethal effects occurred as well in embryos protected against UV-B in the exposure chambers, but were not found in the dark controls. The impact of low dosages of UV-B on the buoyancy of pelagic fish embryos might indicate an important ecological threat and deserves further studies.