Did genome duplication drive the origin of teleosts? A comparative study of diversification in ray-finned fishes

Background  

One of the main explanations for the stunning diversity of teleost fishes (~29,000 species, nearly half of all vertebrates) is that a fish-specific whole-genome duplication event (FSGD) in the ancestor to teleosts triggered their subsequent radiation. However, one critical assumption of this hypothesis, that diversification rates in teleosts increased soon after the acquisition of a duplicated genome, has never been tested.     

Integral valorization of two legumes by autohydrolysis and organosolv delignification

Two woody legumes species (Chamaecytisus proliferus L.F. ssp. palmensis and Leucaena diversifolia) were evaluated for integrally exploitation. The raw material was subjected to autohydrolysis under variable operating conditions which provided a liquid phase rich in hemicellulose oligomers and a solid phase that was used to obtain cellulose pulp and paper sheets by using organosolv procedures. The chemical properties of both C. proliferus and L. diversifolia allow their integral exploitation by using a hydrothermal treatment prior to their organosolv pulping with ethanol. The pulp yields obtained are quite high (40.3% for L. diversifolia and 58.2% for C. proliferus), and so are the sugar concentrations in the liquors from the thermal pretreatment (viz. 16.1 and 20.0 g oligomers/l in C. proliferus and L. diversifolia, respectively, and 1.5 and 1.1g xylose/l, respectively, in the two raw materials). The strength-related properties of the paper sheets obtained are acceptable (tensile index 7.76 and 10.77 kN m/kg for C. proliferus and L. diversifolia, respectively and kappa index 31 and 12.5 for C. proliferus and L. diversifolia, respectively), but somewhat worse than those provided by other raw materials such as eucalyptus; however, they can be improved by mechanical refining of the pulp.     

Pilot clinical trial of type 1 dendritic cells loaded with autologous tumor lysates combined with GM-CSF, pegylated IFN, and cyclophosphamide for metastatic cancer patients

Twenty-four patients with metastatic cancer received two cycles of four daily immunizations with monocyte-derived dendritic cells (DC). DC were incubated with preheated autologous tumor lysate and subsequently with IFN-α, TNF-α, and polyinosinic:polycytidylic acid to attain type 1 maturation. One DC dose was delivered intranodally, under ultrasound control, and the rest intradermally in the opposite thigh. Cyclophosphamide (day -7), GM-CSF (days 1-4), and pegIFN alpha-2a (days 1 and 8) completed each treatment cycle. Pretreatment with cyclophosphamide decreased regulatory T cells to levels observed in healthy subjects both in terms of percentage and in absolute counts in peripheral blood. Treatment induced sustained elevations of IL-12 in serum that correlated with the output of IL-12p70 from cultured DC from each individual. NK activity in peripheral blood was increased and also correlated with the serum concentration of IL-12p70 in each patient. Circulating endothelial cells decreased in 17 of 18 patients, and circulating tumor cells markedly dropped in 6 of 19 cases. IFN-γ-ELISPOT responses to DC plus tumor lysate were observed in 4 of 11 evaluated cases. Tracing DC migration with [(111)In] scintigraphy showed that intranodal injections reached deeper lymphatic chains in 61% of patients, whereas with intradermal injections a small fraction of injected DC was almost constantly shown to reach draining inguinal lymph nodes. Five patients experienced disease stabilization, but no objective responses were documented. This combinatorial immunotherapy strategy is safe and feasible, and its immunobiological effects suggest potential activity in patients with minimal residual disease. A randomized trial exploring this hypothesis is currently ongoing.     

An enzyme-coupled continuous spectrophotometric assay for S-adenosylmethionine-dependent methyltransferases

Modification of small molecules and proteins by methyltransferases affects a wide range of biological processes. Here, we report an enzyme-coupled continuous spectrophotometric assay to quantitatively characterize S-adenosyl-L-methionine (AdoMet/SAM)-dependent methyltransferase activity. In this assay, S-adenosyl-L-homocysteine (AdoHcy/SAH), the transmethylation product of AdoMet-dependent methyltransferases, is hydrolyzed to S-ribosylhomocysteine and adenine by recombinant S-adenosylhomocysteine/5'-methylthioadenosine nucleosidase (SAHN/MTAN, EC 3.2.2.9). Subsequently, adenine generated from AdoHcy is further hydrolyzed to hypoxanthine and ammonia by recombinant adenine deaminase (EC 3.5.4.2). This deamination is associated with a decrease in absorbance at 265 nm that can be monitored continuously. Coupling enzymes are recombinant and easily purified. The utility of this assay was shown using recombinant rat protein arginine N-methyltransferase 1 (PRMT1, EC 2.1.1.125), which catalyzes the mono- and dimethylation of guanidino nitrogens of arginine residues in select proteins. Using this assay, the kinetic parameters of PRMT1 with three synthetic peptides were determined. An advantage of this assay is the destruction of AdoHcy by AdoHcy nucleosidase, which alleviates AdoHcy product feedback inhibition of S-adenosylmethionine-dependent methyltransferases. Finally, this method may be used to assay other enzymes that produce AdoHcy, 5'-methylthioadenosine, or compounds that can be cleaved by AdoHcy nucleosidase.     

Ligand-dependent structural changes and limited proteolysis of Escherichia coli phosphofructokinase-2

An isoform (rhesus UGT1A01) orthologus to the human UGT1A1 was cloned and sequenced from female rhesus monkey liver cDNA using primers designed from the human nucleotide sequences. Open reading frame analysis of the PCR-generated product encodes a 533-amino acid protein with a proposed 27-residue signal peptide. Nucleotide sequence comparison of rhesus UGT1A01 to other rhesus UGT1A isoforms detected a single-transition mutation at nucleotide 1520 (T-->C), resulting in a neutral F to S substitution at position 507. Rhesus UGT1A01 was greater than 99 and 95% identical to cynomolgus UGT1A01 and human UGT1A1, respectively. The rhesus UGT1A01 was expressed in HK-293 cells for functional analysis. Catalytic activity of UGT1A01 was determined with 7-hydroxy-4-(trifluoromethyl)-coumarin and more specific human UGT1A1 substrates (1-naphthol, beta-estradiol, 17 alpha-ethinylestradiol, and bilirubin). Expression of UGT1A01 protein was also detected by a Western blot utilizing a polyclonal antibody developed against the human UGT1A family.     

Stress in Yucatan spider monkeys: effects of environmental conditions on fecal cortisol levels in wild and captive populations

In the Yucatán Peninsula, spider monkeys Ateles geoffroyi yucatanensis are generally found in two contrasting conditions: large tracts of conserved forest or small fragments surrounded by human populations. In the present study, we analyzed fecal cortisol levels of spider monkeys to investigate whether environmental conditions have an influence on stress; specifically, we compared fecal cortisol across individuals living in conserved forests, fragmented forests and captive conditions (zoos and pets). Radioimmunoanalysis of fecal samples from 121 individuals indicated significant differences in mean cortisol for A. g. yucatanensis based on habitat type, with the lowest levels found in the conserved forest condition. The higher cortisol levels in both fragmented forest populations and in captive individuals may be the result of metabolic and behavioral stress. The mean male fecal cortisol concentration was significantly higher than that of females, and the fecal cortisol concentration was higher in the dry season compared with the wet season in a conserved habitat. Therefore, we emphasize the importance of considering sex and seasonality when monitoring fecal cortisol concentrations of spider monkeys, and more generally of frugivores, as they face a seasonal variation in food availability. Finally, our results suggest that forest fragmentation may create long-term stressors for spider monkeys, affecting the viability of populations living under such conditions.