Sequence analysis of phosphoserine-containing peptides. Modification for picomolar sensitivity

Sequencing of phosphoserine-containing peptides yields normally no identifiable PTH-derivatives at those positions where phosphoserine is located. Here a new method is described which allows identification of the position of phosphoserine by chemical modification just before sequence analysis. In a one-step microbatch reaction, phosphoserine present in the intact peptide can be transformed quantitatively into stable derivatives such as beta-methylaminoalanine (MAA), S-ethanolcysteine or S-ethylcysteine. These derivatives are detectable during microsequencing with less than 100 pmol peptide using an Applied Biosystems gas-phase sequencer equipped with an on-line PTH amino acid analyzer.     

Mutagenic potential of permethrin in the Drosophila melanogaster (Diptera: Drosophilidae) sex-linked recessive lethal test

When permethrin was tested for mutagenicity in Drosophila melanogaster Meigen with the sex-linked recessive lethal test, it was nonmutagenic under conditions of this study. The frequencies of spontaneous mutation for permethrin and the negative control were 0.135% and 0.133%, respectively; the spontaneous mutation frequency for positive control was 12.6%. The difference between the mutation frequency of permethrin and the negative control was not significant.     

Colonization rates and community structure of diatoms on three different rock substrata in a lotic system

Diatom assemblages were monitored at weekly intervals over a 5 week period on Verde limestone, Supai sandstone, and Andesitic basalt substrata in a mountain stream in northern Arizona, U.S.A. Density, Shannon-Weiner diversity, evenness, and community similarity (SIMI) were used to compare colonization patterns and community structure between individual substratum types. Average standing crop values were nearly two-fold higher on sandstone than on either basalt or limestone substrata after the first week of the study. It is proposed that differences in micro-surface features between substrata and possibly the rate of substratum solubilization may cause these differences in density early in the colonization period. Following the initial week, standing crop and community structure were significantly similar on all substrata for the remainder of the study period. Maximum densities were attained by the third week and remained relatively constant on all substrata for the remainder of the study.

SEM micrographs demonstrated that surfaces of submerged substrata in streams are modified after the first week by the accumulation of organic aggregates. The establishment of an “organic matrix” early in the colonization process may provide relatively similar attachment surfaces for microbial invasion. This appears to reduce the initial microtopographic differences displayed by substrata and allows for a more uniform colonization pattern.     

Tea catechins’ affinity for human cannabinoid receptors

Among the many known health benefits of tea catechins count anti-inflammatory and neuroprotective activities, as well as effects on the regulation of food intake. Here we address cannabimimetic bioactivity of catechin derivatives occurring in tea leaves as a possible cellular effector of these functionalities. Competitive radioligand binding assays using recombinant human cannabinoid receptors expressed in Chem-1 and CHO cells identified (–)-epigallocatechin-3-O-gallate, EGCG (K_i=33.6 μM), (–)-epigallocatechin, EGC (K_i=35.7 μM), and (–)-epicatechin-3-O-gallate, ECG (K_i=47.3 μM) as ligands with moderate affinity for type 1 cannabinoid receptors, CB1. Binding to CB2 was weaker with inhibition constants exceeding 50 μM for EGC and ECG. The epimers (+)-catechin and (–)-epicatechin exhibited negligible affinities for both CB1 and CB2. It can be concluded that central nervous cannabinoid receptors may be targeted by selected tea catechins but signaling via peripheral type receptors is less likely to play a major role in vivo.     

Ultrafast vascular strain compounding using plane wave transmission

Deformations of the atherosclerotic vascular wall induced by the pulsating blood can be estimated using ultrasound strain imaging. Because these deformations indirectly provide information on mechanical plaque composition, strain imaging is a promising technique for differentiating between stable and vulnerable atherosclerotic plaques. This paper first explains 1-D radial strain estimation as applied intravascularly in coronary arteries. Next, recent methods for noninvasive vascular strain estimation in a transverse imaging plane are discussed. Finally, a compounding technique that our group recently developed is explained. This technique combines motion estimates of subsequently acquired focused ultrasound images obtained at various insonification angles. However, because the artery moves and deforms during the multi-angle acquisition, errors are introduced when compounding. Recent advances in computational power have enabled plane wave ultrasound acquisition, which allows 100 times faster image acquisition and thus might resolve the motion artifacts. In this paper the performance of strain imaging using plane wave compounding is investigated using simulations of an artery with a vulnerable plaque and experimental data of a two-layered vessel phantom. The results show that plane wave compounding outperforms 0° focused strain imaging. For the simulations, the root mean squared error reduced by 66% and 50% for radial and circumferential strain, respectively. For the experiments, the elastographic signal-to-noise and contrast-to-noise ratio (SNR_e and CNR_e) increased with 2.1 dB and 3.7 dB radially, and 5.6 dB and 16.2 dB circumferentially. Because of the high frame rate, the plane wave compounding technique can even be further optimized and extended to 3D in future.     

The Sphingolipid Receptor S1PR2 Is a Receptor for Nogo-A Repressing Synaptic Plasticity

Nogo-A is a membrane protein of the central nervous system (CNS) restricting neurite growth and synaptic plasticity via two extracellular domains: Nogo-66 and Nogo-A-Δ20. Receptors transducing Nogo-A-Δ20 signaling remained elusive so far. Here we identify the G protein-coupled receptor (GPCR) sphingosine 1-phosphate receptor 2 (S1PR2) as a Nogo-A-Δ20-specific receptor. Nogo-A-Δ20 binds S1PR2 on sites distinct from the pocket of the sphingolipid sphingosine 1-phosphate (S1P) and signals via the G protein G₁₃, the Rho GEF LARG, and RhoA. Deleting or blocking S1PR2 counteracts Nogo-A-Δ20- and myelin-mediated inhibition of neurite outgrowth and cell spreading. Blockade of S1PR2 strongly enhances long-term potentiation (LTP) in the hippocampus of wild-type but not Nogo-A^−/− mice, indicating a repressor function of the Nogo-A/S1PR2 axis in synaptic plasticity. A similar increase in LTP was also observed in the motor cortex after S1PR2 blockade. We propose a novel signaling model in which a GPCR functions as a receptor for two structurally unrelated ligands, a membrane protein and a sphingolipid. Elucidating Nogo-A/S1PR2 signaling platforms will provide new insights into regulation of synaptic plasticity.     

Interferon Regulatory Factor-1 Protects from Fatal Neurotropic Infection with Vesicular Stomatitis Virus by Specific Inhibition of Viral Replication in Neurons

The innate immune system protects cells against invading viral pathogens by the auto- and paracrine action of type I interferon (IFN). In addition, the interferon regulatory factor (IRF)-1 can induce alternative intrinsic antiviral responses. Although both, type I IFN and IRF-1 mediate their antiviral action by inducing overlapping subsets of IFN stimulated genes, the functional role of this alternative antiviral action of IRF-1 in context of viral infections in vivo remains unknown. Here, we report that IRF-1 is essential to counteract the neuropathology of vesicular stomatitis virus (VSV). IFN- and IRF-1-dependent antiviral responses act sequentially to create a layered antiviral protection program against VSV infections. Upon intranasal infection, VSV is cleared in the presence or absence of IRF-1 in peripheral organs, but IRF-1^−/− mice continue to propagate the virus in the brain and succumb. Although rapid IFN induction leads to a decline in VSV titers early on, viral replication is re-enforced in the brains of IRF-1^−/− mice. While IFN provides short-term protection, IRF-1 is induced with delayed kinetics and controls viral replication at later stages of infection. IRF-1 has no influence on viral entry but inhibits viral replication in neurons and viral spread through the CNS, which leads to fatal inflammatory responses in the CNS. These data support a temporal, non-redundant antiviral function of type I IFN and IRF-1, the latter playing a crucial role in late time points of VSV infection in the brain.     

The Distribution of Metals in Soils and Pore Water at Three U.S. Military Training Facilities

Small arms firing ranges at military training facilities can have enormous heavy metal burdens (percent level) in soils. Currently there are few published works that quantify the metal content of soils and waters at military installations or speculate on the potential for migration of these contaminants into groundwater. This article documents metals in soils and waters at nine small arms training ranges at three military installations in the U.S. Soil samples were collected from the surface and shallow subsurface. The results demonstrated that lead, antimony, copper, and zinc were the principal contaminants of interest and mapping a site's lead and copper surface distributions would adequately define the extent of impacted soil. Lower metal concentrations at three of the ranges reflected previous remediation by means of physical separation and mechanical removal of metallic fragments followed by fixation treatment with Maectite^TM. Except for the treated ranges where mixing had occurred, subsurface soil samples indicated limited vertical migration. Several of the ranges were also monitored for trace element migration in the vadose zone by means of suction-cup lysimeters. This pore-water sampling indicated ceramic suction-cup lysimeters are useful for assessing relative concentrations but require care in evaluation because of potential sorption losses. Monitoring of soil water at ranges should include antimony and zinc; the former because, in contrast to the other metals, it is typically soluble in an anionic form, and the latter because of its greater solubility and mobility.     

Current and substrate preferences of benthic invertebrates in the rivers of the Hindu Kush-Himalayan region as indicators of hydromorphological degradation

The study introduces an approach to obtaining information about the preferences of benthic invertebrates for substrate and current velocity in a region with little prior knowledge of benthic invertebrates. These preferences are then used for river assessment. Substrate-specific sampling of 271 reference sites was conducted in lower mountainous and lowland areas of the Hindu Kush-Himalaya region. Statistical analysis revealed significant preferences for substrate type and current velocity for 50 taxa of Ephemeroptera, Plecoptera, Trichoptera, Coleoptera, Diptera, Odonata, Mollusca, and Oligochaeta. A 20-point system was developed to assign scores for substrate and current preferences. Scores from seven taxa of Ephemeroptera and Trichoptera revealed low ecological potential in response to habitat alteration. These data were used to develop four preference metrics. The Lithal metric is composed of 34 taxa with significant preferences for stony substrates (fine gravel to bedrock size). The Lithophile metric contains 21 taxa with strong statistical links to stony substrates, which were also found on other substrates. The Lithobiont metric consists of 13 taxa exclusively found on stones. The Lotic metric consists of 11 taxa with significant preferences for moderate-to-fast current velocities. Multi-habitat sampling was conducted at 181 sites reflecting a hydromorphological gradient. The Mann–Whitney U test and box-and-whisker plots were applied to test the relationship of the new metrics to hydromorphological stress. Of the four new metrics, the Lithal, Lithophile, and Lotic were able to detect impacts of hydromorphological degradation.