The Effect of N-Terminal Cyclization on the Function of the HIV Entry Inhibitor 5P12-RANTES.

Despite effective treatment for those living with Human Immunodeficiency Virus (HIV), there are still two million new infections each year. Protein-based HIV entry inhibitors, being highly effective and specific, could be used to protect people from initial infection. One of the most promising of these for clinical use is 5P12-RANTES, a variant of the chemokine RANTES/CCL5. The N-terminal amino acid of 5P12-RANTES is glutamine (Gln; called Q0), a residue that is prone to spontaneous cyclization when at the N-terminus of a protein. It is not known how this cyclization affects the potency of the inhibitor or whether cyclization is necessary for the function of the protein, although the N-terminal region of RANTES has been shown to be critical for receptor interactions, with even small changes having a large effect. We have studied the kinetics of cyclization of 5P12-RANTES as well as N-terminal variations of the protein that either produce an identical cyclized terminus (Glu0) or that cannot similarly cyclize (Asn0, Phe0, Ile0, and Leu0). We find that the half life for N-terminal cyclization of Gln is roughly 20 h at pH 7.3 at 37 °C. However, our results show that cyclization is not necessary for the potency of this protein and that several replacement terminal amino acids produce nearly-equally potent HIV inhibitors while remaining CC chemokine receptor 5 (CCR5) antagonists. This work has ramifications for the production of active 5P12-RANTES for use in the clinic, while also opening the possibility of developing other inhibitors by varying the N-terminus of the protein

Injury Standards in Section 337 Investigations

In this article the authors analyze the impact of the Trade Act of 1974 on Section 337 cases. They focus on the standards the International Trade Commission (ITC) uses in making its injury determinations in Section 337 cases. The authors also address the limited nature of judicial review over final ITC determinations, and conclude that the enhanced role of the ITC in Section 337 proceedings brought about by the Trade Act of 1974 creates a need for private litigants to build a thorough record on the issue of actual or incipient injury in proceedings before the IT

Injury Standards in Section 337 Investigations

In this article the authors analyze the impact of the Trade Act of 1974 on Section 337 cases. They focus on the standards the International Trade Commission (ITC) uses in making its injury determinations in Section 337 cases. The authors also address the limited nature of judicial review over final ITC determinations, and conclude that the enhanced role of the ITC in Section 337 proceedings brought about by the Trade Act of 1974 creates a need for private litigants to build a thorough record on the issue of actual or incipient injury in proceedings before the IT

Implementation of mean-timing and subsequent logic functions on an FPGA

This article describes the implementation of a mean-timer and coincidence logic on a Virtex-5 FPGA for trigger purposes in a particle physics experiment. The novel feature is that the mean-timing and the coincidence logic are not synchronized with a clock which allows for a higher resolution of approximately 400 ps, not limited by a clock frequency.Comment: 15 pages, 11 figure

Development of a 1 GS/s high-resolution transient recorder

With present-day detectors in high energy physics one is often faced with short analog pulses of a few nanoseconds length which may cover large dynamic ranges. In many experiments both amplitude and timing information have to be measured with high accuracy. Additionally, the data rate per readout channel can reach several MHz, which makes high demands on the separation of pile-up pulses. For such applications we have built the GANDALF transient recorder with a resolution of 12bit@1GS/s and an analog bandwidth of 500 MHz. Signals are digitized and processed by fast algorithms to extract pulse arrival times and amplitudes in real-time and to generate experiment trigger signals. With up to 16 analog channels, deep memories and a high data rate interface, this 6U-VME64x/VXS module is not only a dead-time free digitization unit but also has huge numerical capabilities provided by the implementation of a Virtex5-SXT FPGA. Fast algorithms implemented in the FPGA may be used to disentangle possible pile-up pulses and determine timing information from sampled pulse shapes with a time resolution in the picosecond range. Recently the application spectrum has been extended by implementing a 128-channel time-to-digital converter inside the FPGA and an appropriate input mezzanine card

Biochemistry and functional aspects of human glandular kallikreins

Human urinary kallikrein was purified by gel filtration on Sephacryl S-200 and affinity chromatography on aprotinin-Sepharose, followed by ion exchange chromatography on DEAE-Sepharose. In dodecylsulfate gel electrophoresis two protein bands with molecular weights of 41,000 and 34,000 were separated. The amino acid composition and the carbohydrate content of the kallikrein preparation were determined; isoleucine was identified as the only aminoterminal amino acid. The bimolecular velocity constant for the inhibition by diisopropyl fluorophosphate was determined as 9±2 l mol–1 min–1. The hydrolysis of a number of substrates was investigated and AcPheArgOEt was found to be the most sensitive substrate for human urinary kallikrein. Using this substrate an assay method for kallikrein in human urine was developed. It was shown by radioimmunoassay that pig pancreatic kallikrein can be absorbed in the rat intestinal tract. Furthermore, in dogs the renal excretion of glandular kallikrein from blood was demonstrated by radioimmunological methods

Highlighting continued uncertainty in global land cover maps for the user community

In the last 10 years a number of new global datasets have been created and new, more sophisticated alorithms have been designed to classify land cover. GlobCover and MODIS v.5 are the most recent global land cover products available, where GlobCover (300 m) has the finest spatial resolution of other comparable products such as MODIS v.5 (50 m) and GLC-2000 (1 km). This letter shows that the thematic accuracy in the cropland domain has decreased when comparing these two latest products. This disagreement is also evident spatially when examining maps of cropland and forest disargeement between GLC-2000, MODIS and GlobCover. The analysis highlights the continued uncertainty surrounding these prducts, with a combined forest and cropland disagreement of 893 Mha (GlobCover versus MODIS v.5). This letter suggests that data sharing efforts and the provision of more 'in situ' data for training, calibration and validation are very important conditions for improving future global land cover products

The Longitudinal Polarimeter at HERA

The design, construction and operation of a Compton back-scattering laser polarimeter at the HERA storage ring at DESY are described. The device measures the longitudinal polarization of the electron beam between the spin rotators at the HERMES experiment with a fractional systematic uncertainty of 1.6%. A measurement of the beam polarization to an absolute statistical precision of 0.01 requires typically one minute when the device is operated in the multi-photon mode. The polarimeter also measures the polarization of each individual electron bunch to an absolute statistical precision of 0.06 in approximately five minutes. It was found that colliding and non-colliding bunches can have substantially different polarizations. This information is important to the collider experiments H1 and ZEUS for their future longitudinally polarized electron program because those experiments use the colliding bunches only.Comment: 21 pages (Latex), 14 figures (EPS