Quantitative PCR method to detect a 13-kb deletion in the MURR1 gene associated with copper toxicosis and HIV-1 replication

The recently discovered locus for copper toxicosis (CT) in Bedlington terriers (BT) has a 13-kb deletion enveloping the 187-bp exon-2 of the MURR1 gene. This MURR1 gene is not only involved with biliary copper excretion but also associated with HIV-1 replication. The microsatellite C04107 lying in an intron of the MURR1 gene is highly associated with the disease but shows haplotype diversity. The only solid molecular test for the disease is by showing the deletion in exon-2 in cDNA in liver tissue; this test is not robust on RNA from peripheral leukocytes because of their low MURR1 expression level. Because of these drawbacks, we developed a new quantitative PCR (Q-PCR) protocol. Here we show that the MURR1 exon-2/exon-3 ratio measured by Q-PCR on genomic DNA correlates perfectly with the microsatellite marker and with RT-PCR data from blood samples, buccal swabs, and liver biopsies. In view of the important role of MURR1 in cells of many tissues, this new test has a wide range of applications in comparative biomedical research. Furthermore, Q-PCR on DNA may be a new tool in general to analyze mutations that cannot be approached by standard methods.Robert P. Favier and Bart Spee contributed equally to this work.     

Neutrophil inhibition with L-selectin-directed MAb improves or worsens survival dependent on the route but not severity of infection in a rat sepsis model.

Both route and severity of infection may influence immunomodulator agents in sepsis. We studied the effect of each variable on HRL-3, an L-selectin-directed MAb that inhibits neutrophil function, in a rat sepsis model. Animals (n = 800) were randomized to be treated with either HRL-3 or placebo and to receive Escherichia coli either intravenously (IV) or intrabronchially (IB) in doses producing low or high mortality rates. Animals received antibiotics and were observed for 168 h. Route but not dose of E. coli altered the effects HRL-3 on mortality rate (mean hazards ratio +/- SE). With IV E. coli, compared with control, HRL-3 was beneficial and reduced the hazards ratio both early (0 to 6 h; -0.75 +/- 0.23) and late (6 to 168 h; -0.72 +/- 0.36) (P = 0.001 and 0.04, respectively, over all E. coli doses). In contrast, with IB E. coli HRL-3 reduced the hazards ratio early (-1.1 +/- 0.36) but worsened it late (0.87 +/- 0.23) (P = 0.002 for both effects over all E. coli doses) in patterns significantly different from IV E. coli (P < 0.0001). Compared with control, although HRL-3 did not alter lung neutrophil numbers or injury score at 6 or 168 h with IV E. coli (P = ns for all), it reduced both early and increased them late with IB E. coli (P 相似文献   

Spectroscopy and photophysics of flavin-related compounds: 3-benzyl-lumiflavin.

Molecular structure, spectroscopic and photophysical data for the singlet state of 3-benzyl-lumiflavin in different solvents are presented. Theoretical studies concerning singlet-singlet and triplet-triplet excitation energies were carried out using time-dependent density functional theory (TD-DFT) calculations. These predictions are in good agreement with the experimental results, which reflect the solvent interactions. All the observable singlet-singlet transitions have pi-pi* character. The title compound appears to be an efficient sensitizer of the production of singlet oxygen (phi(Delta)= 0.53). The crystal structure of 3-benzyl-lumiflavin is also presented, along with its solid-state photophysical data.     

Quantitative floral phenology at the landscape scale: Is a comparative spatio-temporal description of ‘‘flowering landscapes’’ possible?

Entomophilous flowers form the food resources for insect pollinators. Many pollinator species forage at the landscape scale and depend on floral resources that are highly variable in space and time. We present a general model approach in which the floral resources of plant communities are estimated by the floral phenology and the cover of entomophilous plant species. We applied this landscape model in a case study for three landscape sections (1.5–2.2 km²) with strongly differing land-use patterns. The comparison between a conservation area and two agricultural landscapes shows extreme differences in the quantities and in the course of floral resources.

In a stepwise simplification of the landscape model we tested the effects of input data with lower spatio-temporal resolution. Even if input data for floral phenology and vegetation have a low resolution, the landscape model allows a ranking of landscape-specific floral resource potentials. The results of the case study encourage the use of landscape models to estimate floral resource potentials. The assessment of floral resource potentials may help to define this essential landscape quality for evaluation in practical nature conservation.