The adjuvant effect of Sophy β-glucan to the antibody response in poultry immunized by the avian influenza A H5N1 and H5N2 vaccines

Avian influenza virus vaccines produced in oil-emulsified inactivated form with antigen content of at least 160 hemagglutinin units (HAU) induced immunity in birds. However, in addition to enhancing the effect of the adjuvant(s), other additional supplemented biological compounds included in inactivated vaccines could produce higher levels of antibody. We examined in chickens, Vietnamese ducks, and muscovy ducks the adjuvant effect of Sophy β-glucan (SBG), a β-1,3-1,6 glucan produced by the black yeast Aureobasidium pollulans strain AF0-202, when administered with an avian influenza H5 subtype vaccine. In Experiment 1, 40 chickens (ISA Brown hybrid), allocated to four groups of ten each, were immunized with Oil-H5N1(VN), Oil-H5N1(CN), Oil-H5N2(CN), and saline (control group), respectively. In Experiment 2, chickens (ISA Brown hybrid), muscovy ducks (French hybrid), and Vietnamese ducks (indigenous Vietnamese) were used to further assess the effect of SBG on immunogenicity of the Oil-H5N1(VN) Vietnamese vaccine. ELISA and hemagglutination inhibition (HI) assays were used to assess the antibody response. The H5 subtype vaccines initiated significantly higher immune responses in the animals dosed with SBG, with 1.0-1.5 log2 higher HI titers and 10-20% ELISA seroconversion, compared with those not dosed with β-glucan. Notably, some of the animals dosed with SBG induced HI titers higher than 9.0 log2 following boosting immunization. Taken together, our serial studies indicated that SBG is a potential effector, such as enhancing the immune response to the H5 vaccines tested.     

Letter to the Editor: Backbone 1H, 15N and 13C Assignments for the Subunit a of the E. Coli ATP Synthase

The structure of the 30 KDa subunit a of the membrane component (F(0)) of E. coli ATP synthase is investigated in a mixture of chloroform, methanol and water, a solvent previously used for solving the structure of another integral membrane protein, subunit c. Near complete backbone chemical shift assignments were made from a set of TROSY experiments including HNCO, HNCA, HN(CA)CB, HN(CO)CACB and 4D HNCOCA and HNCACO. Secondary structure of subunit a was predicted from the backbone chemical shifts using TALOS program. The protein was found to consist of multiple elongated alpha-helical segments. This finding is generally consistent with previous predictions of multiple transmembrane alpha-helices in this polytopic protein.     

Partial Protection of Carbohydrate Derivatives. Part 91. Equilibration between 2′, 5′- and 3′, 5′-DI-O-Benzoyladenosine Derivatives Substituted at the N 6-Position,on Silica Gel

Abstract

Equilibration between 2′, 5′- and 3′, 5′-di-O-benzoyladenosine derivatives on Wakogel C-300 and Merck 60 silica gel was effectively induced to give a mixture predominantly containing the latter; further potential factors involved were also investigated.     

Synthesis,structures and characterization of the dinuclear nickel(II) complexes containing N,N,N′,N′-tetrakis[(1-ethyl-2-benzimidazolyl)methyl]-2-hydroxy-1,3-diaminopropane and an azide ion

Two dinuclear nickel(II) complexes, [Ni₂(L-Et)(N₃)(H₂O)₃](NO₃)₂ · 2H₂O (1) and [Ni₂(L-Et)(μ-1,3-N₃)(H₂O)₂](NO₃)₂ · 4H₂O (2) containing (HL-Et = N,N,N′,N′-tetrakis[(1-ethyl-2-benzimidazolyl)methyl]-2-hydroxy-1,3-diaminopropane), have been synthesized and characterized by their IR and UV–Vis spectra and magnetic susceptibilities. The crystal structures of [Ni₂(L-Et)(N₃)(H₂O)₃](NO₃)₂ · CH₃OH (1′) and [Ni₂(L-Et)(μ-1,3-N₃)(H₂O)₂](NO₃)₂ · 2C₂H₅OH (2′) similar to 1 and 2 were determined by X-ray crystallography. In 1′, the two nickel(II) ions are bridged by only an alkoxo group of L-Et⁻, while an azido and an alkoxo connect two nickel(II) ions in 2′. Magnetic susceptibility measurements (2–300 K) showed a weak ferromagnetic exchange coupling between the two nickel(II) ions (2J = 10.1 cm⁻¹) for 1. On the other hand, antiferromagnetic interactions were observed for 2 (2J = −15.8 cm⁻¹).     

The Idea Is Good,but…: Failure to Replicate Associations of Oxytocinergic Polymorphisms with Face-Inversion in the N170

Background

In event-related potentials, the N170 manifests itself especially in reaction to faces. In the healthy population, face-inversion leads to stronger negative amplitudes and prolonged latencies of the N170, effects not being present in patients with autism-spectrum-disorder (ASD). ASD has frequently been associated with differences in oxytocinergic neurotransmission. This ERP-study aimed to investigate the face-inversion effect in association with oxytocinergic candidate genes. It was expected that risk-allele-carriers of the oxytocin-receptor-gene-polymorphism (rs53576) and of CD38 (rs379863) responded similar to upright and inverted faces as persons with ASD. Additionally, reactions to different facial emotional expressions were studied. As there have been difficulties with replications of those molecular genetic association studies, we aimed to replicate our findings in a second study.

Method

Seventy-two male subjects in the first-, and seventy-eight young male subjects in the replication-study conducted a face-inversion-paradigm, while recording EEG. DNA was extracted from buccal cells.

Results

Results revealed stronger N170-amplitudes and longer latencies in reaction to inverted faces in comparison to upright ones. Furthermore, effects of emotion on N170 were evident. Those effects were present in the first and in the second study. Whereas we found molecular-genetic associations of oxytocinergic polymorphisms with the N170 in the first study, we failed to do so in the replication sample.

Conclusion

Results indicate that a deeper theoretical understanding of this research-field is needed, in order to generate possible explanations for these findings. Results, furthermore, support the hypotheses that success of reproducibility is correlated with strength of lower original p-values and larger effect sizes in the original study.     

Sociology is an applied science,and the application is social change: a contribution to the discussion of Aldon Morris’ The Scholar Denied

I argue that W.E.B. Du Bois’ expulsion from academic sociology at the beginning of the twentieth century was not only animated by the gatekeepers’ desire to maintain academe as a whites-only domain. It also constituted an active effort to defend Social Darwinist dogma, which dominated (white) social science at that time, from the formidable challenge of a resourceful and scientifically superior perspective. The successful exclusion of Du Bois (and his growing legion of colleagues) allowed sociology to drift into functionalist dogma, with its immutable hierarchy and denial of sociology’s role in facilitating social change. The exclusion of Du Boisean analysis, which placed human agency – and especially subaltern groups – at the centre of social change – rendered sociological analysis irrelevant to addressing social problems and social justice, until the Civil Rights Movement in America broke down the walls of the sociological ghetto, allowing it to access the rich Du Boisean perspective.     

The α2L111R,N122D Isoform of the Na,K-ATPase Expressed in HeLa Cells Does Not Undergo an Adipocyte-like Increase in Activity in Response to Insulin

In the rat adipocyte, insulin increases potassium uptake by a preferential activation of the α2 isoform of the Na,K-ATPase. The question under consideration here is whether expression of the α2 isoform is sufficient to replicate its differential activation by insulin. Accordingly, we compared the effect of insulin on the activity of the ouabain resistant rat α1 and α2RD (α2L111R,N122D) isoforms in HeLa cells. In HeLa cells, in contrast to the rat adipocyte, insulin produces an increase of equal magnitude in the rate of⁸⁶Rb⁺/K⁺uptake by the ouabain resistant rat α1 and rat α2RD subunits. We conclude that the mechanism of insulin activation of the α2RD isoform in HeLa cells differs from that of the wild type α2 isoform in the rat adipocyte.     

3′/5′-Regioselectivity of Introduction of the 9-Fluorenyl-Methoxycarbonyl Group to 2′-o-Tetrahydropyran-2-YL-and 2′-O-(4-Methoxytetrahydropyran-4-YL-)-Nucleosides: Useful Intermediates for Solid-Phase-Rna-Synthesis

Abstract

X-ray structure analysis of the more laevorotatory isomers of 2′-O-tetrahydropyranyl-4N-benzoylcytidine (4b) and of 2′-O-tetrahydropyranyluridine (5b) confirmed their chirality at the satellite anomeric centre C2″ to be S. The other diastereomers (4a resp. 5a) exhibited an unexpected reversal of 3′/5′-regio-selectivity when treated with 9-fluorenylmethoxycarbonyl chloride in pyridine. The X-ray crystallographic results form the basis for a mechanistic proposal.     

The 3′-Phosphoadenosine 5′-Phosphosulfate Transporters,PAPST1 and 2, Contribute to the Maintenance and Differentiation of Mouse Embryonic Stem Cells

Recently, we have identified two 3′-phosphoadenosine 5′-phosphosulfate (PAPS) transporters (PAPST1 and PAPST2), which contribute to PAPS transport into the Golgi, in both human and Drosophila. Mutation and RNA interference (RNAi) of the Drosophila PAPST have shown the importance of PAPST-dependent sulfation of carbohydrates and proteins during development. However, the functional roles of PAPST in mammals are largely unknown. Here, we investigated whether PAPST-dependent sulfation is involved in regulating signaling pathways required for the maintenance of mouse embryonic stem cells (mESCs), differentiation into the three germ layers, and neurogenesis. By using a yeast expression system, mouse PAPST1 and PAPST2 proteins were shown to have PAPS transport activity with an apparent K_m value of 1.54 µM or 1.49 µM, respectively. RNAi-mediated knockdown of each PAPST induced the reduction of chondroitin sulfate (CS) chain sulfation as well as heparan sulfate (HS) chain sulfation, and inhibited mESC self-renewal due to defects in several signaling pathways. However, we suggest that these effects were due to reduced HS, not CS, chain sulfation, because knockdown of mouse N-deacetylase/N-sulfotransferase, which catalyzes the first step of HS sulfation, in mESCs gave similar results to those observed in PAPST-knockdown mESCs, but depletion of CS chains did not. On the other hand, during embryoid body formation, PAPST-knockdown mESCs exhibited abnormal differentiation, in particular neurogenesis was promoted, presumably due to the observed defects in BMP, FGF and Wnt signaling. The latter were reduced as a result of the reduction in both HS and CS chain sulfation. We propose that PAPST-dependent sulfation of HS or CS chains, which is regulated developmentally, regulates the extrinsic signaling required for the maintenance and normal differentiation of mESCs.     

Crystal and molecular structure of [N,N′-ethylenebis(methyl-2-amino-1-cyclopentenedithiocarboxylato)]copper(II): A quantitative relationship between tetrahedral distortion and redox potentials in copper N2S2 compounds and the relevance to type I copper(II) centers

The crystal and molecular structure of the copper(II) complex of the N₂S₂ tetradentate ligand, ethylenebis(methyl-2-amino-1-cyclopentenedithiocarboxylate), was solved at room temperature by a single crystal x-ray diffraction study. The complex crystallizes in the orthorhombic space group P2₁2₁2₁ with a = 7.739(1) Å, b = 13.893(2) Å, c = 17.096(3) Å, V = 1838(1) ų, ?_observed = 1.56 g cm^?3 and ?_calculated = 1.57 g cm^?3 for a molecular weight of 434.2, and Z = 4. Diffraction data were collected with a Syntex P$\text{1}$ diffractometer using graphite-monochromatized Cu (λ = 1.5418 Å) radiation. The heavy atoms were located from a Patterson synthesis; all other nonhydrogen atoms were located using difference Fourier techniques, and hydrogen atoms were placed in calculated positions. Final refinement resulted in discrepancy indices of R = 0.067 and goodness of fit of 2.92 for all 995 reflections (5° < 2θ < 100°) greater than three times their standard deviation. The molecules are monomeric and well separated. Bond distances in the two ”halvesldquo; of the ligand are sufficiently different to suggest that different resonance structures exist in each portion. This agrees with the rhombic symmetry displayed by the frozen glass esr spectrum of the compound (_xx ≠ g_yy). The dihedral angle between the planes defined by the CuN₂ and CuS₂ planes is 20.0°, indicating a rather distorted inner coordination sphere. The copper(II)-copper(I) reduction potentials found for this compound and the trimethylene and tetramethylene analogs were determined to be ?1.01, ?0.79, and ?0.64 V respectively. A quantitative relationship between tetrahedral distortion and redox potentials is obtained, and these results are discussed in terms of ”blueldquo; copper(II) sites in proteins. Trends in CuS and CuN bonding patterns in the same three compounds are discussed with regard to the short CuS (cys) bond distance in plastocyani Finally, a brief discussion of the optical spectra of these three compounds, their variation, and their significance with respect to tetrahedral symmetry in copper(II) protein sites is presented.     

Synthesis of the 2′-,3′-Didehydro-2′-,3′-dideoxy and 2′-,3′-Dideoxy Derivatives of 6-Azauridine and a New Route to 2′-,3′-Didehydro-2′-,3′-dideoxy-5-chlorouridine

Abstract

The 5′-O-(4,4′-dimethoxytrityl) and 5′-O-(tert-butyldimethylsilyl) derivatives of 2′-,3′-O-thiocarbonyl-6-azauridine and 2′,3′-O-thiocarbonyl-5-chlorouridine were synthesized from the parent nucleosides by reaction with 4, 4′-dimethoxytrityl chloride and tert-butyldimethylsilyl chloride, respectively, followed by treatment with 1,1′-thiocarbonyldiimidazole. Introduction of a 2′-,3′-double bond into the sugar ring by reaction of the 5′-protected 2′-,3′-O-thionocarbonates with 1, 3-dimethyl-2-phenyl-1, 3, 2-diazaphospholidiine was unsuccessful, but could be accomplished satisfactorily with trimethyl phosphite. Reactions were generally more successful with the 5′-silylated than with the 5′-tritylated nucleosides. Formation of 2′-,3′-O-thiocarbonyl derivatives proceeded in higher yield with 5′-protected 6-azauridines than with the corresponding 5-chlorouridines because of the propensity of the latter to form 2,2′-anhydro derivatives. In the reaction of 5′-O-(tert-butyldimethylsilyl)-2′-,3′-O-thiocarbonyl-6-azauridine with trimethyl phosphite, introduction of the double bond was accompanied by N³-methylation. However this side reaction was not a problem with 5′-O-(tert-butyldimethylsilyl)-2′-, 3′-O-thioarbonyl-5-chlorouridine. Treatment of 5′-O-(tert-butyldimethylsilyl)-2′-, 3′-didehydro-2′-,3′-dideoxy-6-azauridine with tetrabutylammonium fluoride followed by hydrogenation afforded 2′-,3′-dideoxy-6-azauridine. Deprotection of 5′-O-(tert-butyldimethylsilyl)-2′-, 3′-didehydro-2′-,3′-dideoxy-5-chlorouridine yielded 2′-,3′-didehydro-2′-,3′-dide-oxy-5-chlorouridine.