A radioimmunoassay of plasma estradiol-17beta with the use of polyethylene glycol to separate free and antibody-bound hormone

A radioimmunoassay for plasma estradiol-17β was developed using polyethylene glycol to separate free from antibody-bound hormone. Specificity for estradiol-17β was achieved by a modified celite microcolunm procedure in which estradiol was.separated from interfering estrogens, including estrone. Using trace ³H-estradiol to monitor procedural losses, the method was shown to be sensitive and accurate. Intra- and inter-assay coefficient of variation of the method was 8.7 and 10.6%, respectively. Polyethylene glycol used for antibody precipitation appears to be a generally applicable method for steroid hormone radioimmunoassays. The simplicity, precision and rapid analysis, coupled with its lack of time dependence and ease in automation, makes this a convenient and practical method.     

Characterization of Mus musculus Papillomavirus 1 Infection In Situ Reveals an Unusual Pattern of Late Gene Expression and Capsid Protein Localization

Full-length genomic DNA of the recently identified laboratory mouse papillomavirus 1 (MusPV1) was synthesized in vitro and was used to establish and characterize a mouse model of papillomavirus pathobiology. MusPV1 DNA, whether naked or encapsidated by MusPV1 or human papillomavirus 16 (HPV 16) capsids, efficiently induced the outgrowth of papillomas as early as 3 weeks after application to abraded skin on the muzzles and tails of athymic NCr nude mice. High concentrations of virions were extracted from homogenized papillomatous tissues and were serially passaged for >10 generations. Neutralization by L1 antisera confirmed that infectious transmission was capsid mediated. Unexpectedly, the skin of the murine back was much less susceptible to virion-induced papillomas than the muzzle or tail. Although reporter pseudovirions readily transduced the skin of the back, infection with native MusPV1 resulted in less viral genome amplification and gene expression on the back, including reduced expression of the L1 protein and very low expression of the L2 protein, results that imply skin region-specific control of postentry aspects of the viral life cycle. Unexpectedly, L1 protein on the back was predominantly cytoplasmic, while on the tail the abundant L1 was cytoplasmic in the lower epithelial layers and nuclear in the upper layers. Nuclear localization of L1 occurred only in cells that coexpressed the minor capsid protein, L2. The pattern of L1 protein staining in the infected epithelium suggests that L1 expression occurs earlier in the MusPV1 life cycle than in the life cycle of high-risk HPV and that virion assembly is regulated by a previously undescribed mechanism.     

Paradigmatic pragmatism and the politics of diversity

How migration and mobilizations of difference are accommodated at the local level is a burning question. Concepts adopted by local governments and the capacities of cities to formulate and implement these have received increasing attention, but often without examining the ideas and norms that underlie local concepts and practices. This article assesses the hypothesis of local-level pragmatism, which it rejects, and develops the notion of ‘paradigmatic pragmatism’ to characterize how local-level politics address mobilizations of difference. Based on empirical findings from Amsterdam, Antwerp and Leeds, and comparing the content and the implementation of ‘diversity policies’, I argue against a dichotomy of pragmatic vs ideational politics. Instead, these cities draw on a variety of ideas and pragmatically combine them under the header of diversity. This paradigmatic pragmatism invites further research on the effects of potential incompatibilities of immigrant policy ideas.     

Hydrological extremes modulate nutrient dynamics in mediterranean climate streams across different spatial scales

We studied benthic macroinvertebrate communities upstream and downstream of five small reservoirs (surface release in autumn–winters) (north Spain) to assess the effect of flow regulation on structural and functional characteristics of stream ecosystems. We based our approach on the use of structural metrics (density, biomass, richness and diversity) in combination with two functional diversity indices based on biological and ecological traits: FD_PG index, related to species richness, and FD_Q, which incorporates evenness across taxa. Although water physicochemical parameters were unaffected by the reservoirs during the study period (autumn–winter), macroinvertebrate metrics were lower below the dams, with detritivores (shredders and collector-gatherers) being the most affected. The alder leaf breakdown rate estimated by the litter-bag technique was related to the density, biomass, richness, diversity and FD_PG index of shredders, compromising the ecosystem functioning. The most plausible origin for the observed differences in macroinvertebrate metrics between upstream and downstream reaches was the change of the flow regime caused by the impoundments at downstream sites, leading to droughts in summer in those naturally permanently flowing streams. The observed functional diversity loss might reduce the chances of the community to override natural or man-induced fluctuations in their environment with possible repercussions on important ecosystem functions and services.     

Transferability of Material Composition Indicators for Residential Buildings: A Conceptual Approach Based on a German‐Japanese Comparison

Most anthropogenic material stocks and flows are associated with the building sector. Several recent studies have developed material composition indicators (MCIs) suitable for calculating material stocks and flows of the building sector using bottom‐up approaches, which hold great potential to provide information to support resource efficiency policies. A major limitation is the lack of country‐specific MCIs. This study aims to introduce a concept for a better transferability of MCI across different contexts by proposing requirements for defining MCIs and to discuss options and limits of the transferability. We take existing MCIs for residential buildings in Germany and Japan as case studies and make them comparable by applying harmonization methods. Based on that, similarities and differences are systematically identified and discussed, considering their socioeconomic, cultural, technical, and environmental factors. Our results indicate significant limitations to the transferability of MCIs for detached houses, while bigger apartment complexes show greater homogeneity despite the very different environments in which they are constructed. This indicates that while it is possible to assume foreign MCIs as plausible for large constructions, local coefficients need to be estimated for smaller single‐family homes.     

DALDA analogues containing alpha-hydroxymethylamino acids

To evaluate the role of aromatic amino-acids residues, four analogues of the mu-selective opioid peptide agonist DALDA (H-Tyr-D-Arg-Phe-Lys-NH2) containing the amphiphilic, a,a-disubstituted amino acid (R)- or (S)-alpha-hydroxymethyltyrosine (HmTyr) in position 1 and (R)- or (S)-alpha-hydroxymethylphenylalanine (HmPhe) in position 3 of the peptide sequence were synthesized. Only the [(R)-HmPhe3)]DALDA analogue displayed full agonistic activity in both the guinea pig ileum and the mouse vas deferens assays and turned out to be a delta receptor-selective opioid agonist.     

Conversion of delta-, kappa- and mu-receptor selective opioid peptide agonists into delta-, kappa- and mu-selective antagonists

2',6'-Dimethyl substitution of the Tyr(1) residue of opioid agonist peptides and deletion of the positively charged N-terminal amino group or its replacement with a methyl group has recently been shown to represent a general structural modification to convert opioid peptide agonists into antagonists. This conversion requires the syntheses of opioid peptide analogues containing either 3-(2,6-dimethyl-4-hydroxyphenyl)propanoic acid (Dhp) or (2S)-2-methyl-3-(2,6-dimethyl-4-hydroxyphenyl)propanoic acid [(2S)-Mdp] in place of Tyr(1). Using this approach, delta-, kappa- and mu-selective opioid peptide agonist peptides were successfully converted into corresponding delta-, kappa- and mu-selective antagonists, whereby receptor selectivity was often maintained or even improved. Thus, two (2S)-Mdp(1)-analogues of the delta-selective cyclic enkephalin analogue H-Tyr-c[D-Pen-Gly-Phe(pF)-Pen]-Phe-OH turned out to be potent and selective delta antagonists. Most successful was the development of kappa antagonists derived from dynorphin A (Dyn A), including the highly potent and selective kappa-antagonist [(2S)-Mdp(1)]Dyn A(1-11)-NH(2) (dynantin) and the enzymatically stable octapeptide analogue [(2S)-Mdp(1),MeArg(7),D-Leu(8)]Dyn A(1-8)-NH(2). The (2S)-Mdp(1)-analogues of dynorphin B and alpha-neoendorphin also were kappa antagonists and may be useful as pharmacological tools in studies of kappa receptor subtypes. Finally, the Dhp(1)-analogues of the mu-selective cyclic enkephalin analogue H-Tyr-c[N(epsilon ),N(beta)-carbonyl-D-Lys(2),Dap(5)]enkephalinamide and of endomorphin-2 were moderately potent mu opioid antagonists.     

Influence of tidal volume on pulmonary NO release,tissue lipid peroxidation and surfactant phospholipids

Mechanical stress during ventilation may cause or aggravate acute lung injury. This study investigates the influence of low vs. high tidal volume (V(t)) on factors known to play key roles in acute lung injury: nitric oxide release, eNOS and iNOS gene expression, lipid peroxidation (LPO), and surfactant phospholipids (PL). Isolated rabbit lungs were subjected to one of three ventilation patterns for 135 min (V(t)-PEEP): 6 ml/kg-0 cm H(2)O. 12 ml/kg-0 cm H(2)O 6 ml/kg-5 cm H(2)O, 12 ml/kg-0 cm H(2)O, and 6 ml/kg-5 cm H(2)O resulted in comparable peak inspiratory pressure (PIP). This allowed comparing low and high V(t) without dependence on PIP. Ventilatory patterns did not induce changes in pulmonary artery pressure, vascular permeability (K(f,c)), PIP or pulmonary compliance. High V(t) in comparison with both of the low V(t) groups caused an increase in BALF-nitrite (30.6+/-3.0* vs. 21.4+/-2.2 and 16.2+/-3.3 microM), BALF-PL (1110+/-19* vs. 750+/-68 and 634+/-82 microg/ml), and tissue LPO product accumulation (0.62+/-0.051* vs. 0.48+/-0.052 and 0.43+/-0.031 nmol/mg), *P<0.05 each. Perfusate nitrite and BALF-PL composition (assessed by use of 31P-NMR spectroscopy and MALDI-TOF mass spectrometry) did not differ among the groups. High V(t) ventilation reduced eNOS gene expression but did not affect iNOS expression. The increased release of NO and the accumulation of LPO products may represent early lung injury while elevated BALF-PL may reflect distension-induced surfactant secretion.