Characterization of the PCMBS-dependent modification of KCa3.1 channel gating.

Intermediate conductance, calcium-activated potassium channels are gated by the binding of intracellular Ca(2+) to calmodulin, a Ca(2+)-binding protein that is constitutively associated with the C terminus of the channel. Although previous studies indicated that the pore-lining residues along the C-terminal portion of S6 contribute to the activation mechanism, little is known about whether the nonluminal face of S6 contributes to this process. Here we demonstrate that the sulfhydral reagent, parachloromercuribenze sulfonate (PCMBS), modifies an endogenous cysteine residue predicted to have a nonluminal orientation (Cys(276)) along the sixth transmembrane segment (S6). Modification of Cys(276) manipulates the steady-state and kinetic behavior of the channel by shifting the gating equilibrium toward the open state, resulting in a left shift in apparent Ca(2+) affinity and a slowing in the deactivation process. Using a six-state gating scheme, our analysis shows that PCMBS slows the transition between the open state back to the third closed state. Interpreting this result in the context of the steady-state and kinetic data suggests that PCMBS functions to shift the gating equilibrium toward the open state by disrupting channel closing. In an attempt to understand whether the nonluminal face of S6 participates in the activation mechanism, we conducted a partial tryptophan scan of this region. Substituting a tryptophan for Leu(281) recapitulated the effect on the steady-state and kinetic behavior observed with PCMBS. Considering the predicted nonluminal orientation of Cys(276) and Leu(281), a simple physical interpretation of these results is that the nonluminal face of S6 forms a critical interaction surface mediating the transition into the closed conformation, suggesting the nonluminal C-terminal portion of S6 is allosterically coupled to the activation gate

Phonorecords and Forfeiture of Common-Law Copyright in Music

A highly disputed issue surrounding the Copyright Act of 1909 is whether the public distribution and sale of recordings of a musical work constitutes publication. Historically, unless the author followed the Act\u27s formal requirements for obtaining statutory protection, publication injected the musical work irrevocably into the public domain. In a 1995 decision, La Cienega Music Co. v. ZZ Top, the Ninth Circuit unwisely broke from the tradition and common understanding in the music industry by holding that phonorecord distribution is a publication of musical compositions. After examining the history and purpose of the Copyright Act, as well as the legal precedents, this Comment argues that Congress did not intend phonorecords to be capable of publication. Besides being unfair to composers of original works, the court has created a split among the circuits that should be resolved

Geographic Distribution: \u3ci\u3eIguana iguana\u3c/i\u3e (Green Iguana): USA: Florida.

A geographic distribution record for Iguana iguana in Marion County, Florida

Evolutionary history determines how plant productivity responds to phylogenetic diversity and species richness

The relationship between biodiversity and ecosystem function has received a great deal of attention in ecological research and recent results, from re-analyses, suggest that ecosystem function improves with increases in phylogenetic diversity. However, many of these results have been generalized across a range of different species and clades, and plants with different evolutionary histories could display different relationships between biodiversity and ecosystem function. To experimentally test this hypothesis, we manipulated species richness and phylogenetic diversity using 26 species from two subgenera of the genus Eucalyptus (subgenus Eucalyptus and subgenus Symphyomyrtus). We found that plant biomass (a measurement of ecosystem function) sometimes, but not always, responded to increases in species richness and phylogenetic diversity. Specifically, Symphyomyrtus plants showed a positive response while no comparable effect was observed for Eucalyptus plants, showing that responses to biodiversity can vary across different phylogenetic groups. Our results show that the impacts of evolutionary history may complicate the relationship between the diversity of plant communities and plant biomass

Experimental Investigation of Blowing Effects on Turbulent Flow over a Rough Surface

A high Reynolds number turbulent channel flow facility was used to study the combined effects of roughness and flow injection on the mean flow and turbulence characteristics. It was found that the additional momentum injection through the surface enhanced the roughness effects and for the mean flow the effect of blowing was indistinguishable from that of increased roughness. However, for the turbulence statistics, this analogy broke down in that the addition of blowing resulted in behavior which did not follow that predicted by Townsend’s hypothesis. This was observed as a reduction of outer-scaled Reynolds stress well into the outer layer. The reduction in Reynolds stress was accomplished primarily by a suppression of the kinetic energy content associated with large-scale motions, which are believed to be formed at the surface but extend into the outer layer

Bioinformatics analysis of the locus for enterocyte effacement provides novel insights into type-III secretion

BACKGROUND: Like many other pathogens, enterohaemorrhagic and enteropathogenic strains of Escherichia coli employ a type-III secretion system to translocate bacterial effector proteins into host cells, where they then disrupt a range of cellular functions. This system is encoded by the locus for enterocyte effacement. Many of the genes within this locus have been assigned names and functions through homology with the better characterised Ysc-Yop system from Yersinia spp. However, the functions and homologies of many LEE genes remain obscure. RESULTS: We have performed a fresh bioinformatics analysis of the LEE. Using PSI-BLAST we have been able to identify several novel homologies between LEE-encoded and Ysc-Yop-associated proteins: Orf2/YscE, Orf5/YscL, rORF8/EscI, SepQ/YscQ, SepL/YopN-TyeA, CesD2/LcrR. In addition, we highlight homology between EspA and flagellin, and report many new homologues of the chaperone CesT. CONCLUSION: We conclude that the vast majority of LEE-encoded proteins do indeed possess homologues and that homology data can be used in combination with experimental data to make fresh functional predictions

Experimental Investigation of Blowing Effects on Turbulent Flow Over a Rough Surface

Entry of spacecraft into an atmosphere occurs at hypersonic speeds, extremely complex flow fields, with aerothermodynamic effects which can cause the surface to be subjected to extreme heating. It is therefore important to protect the vehicle and its payload using a thermal protection system (TPS). Heat shields, which are an important part of a TPS, can be of either ablative or non-ablative types. For an ablative TPS, the energy is dissipated through surface material charring and ablation, as well as releasing gasses which serve to carry energy away from the TPS and thus reduce the total heat flux into the vehicle

Investigation of Turbulent Structure Modification by Momentum Injection Into Turbulent Flow Over a Rough Surface

Utilizing a turbulent channel flow facility, experiments were performed to determine the modifications made to the temporal and spatial structure of turbulence over a sinusoidally rough surface due to momentum injection through the surface. As expected, when compared to the smooth-walled conditions, it was found that the surface roughness increased wall shear stress, decreased the turbulence energy content near the wall, and shifted that energy content to wavelengths on the order of the roughness elements. The addition of momentum injection was found to produce further modifications analogous to increasing the roughness height by increasing the wall shear stress and further reducing the energy content of the near-wall turbulence while simultaneously shifting it to longer wavelengths

Progress in Lithotripsy Research

Shock wave lithotripsy (SWL) for the non-invasive treatment of kidney stones was introduced in the United States in 1984. SWL virtually eliminated the need for open surgery to remove kidney stones, and it did not take long for physicians and patients to endorse this revolutionary technology. Early reports told of the efficient removal of even the most troublesome stones without apparent complications, and SWL quickly became the "treatment modality of choice." It was not long, however, before concerned physicians began to report the occurrence of adverse effects in SWL, particularly involving vascular trauma and including cases of severe hemorrhage in the kidney and acute renal failure — significant side effects of serious consequence. Researchers quickly recognized the challenge and opportunity to determine the mechanisms of shock wave action in lithotripsy, and in 1988, the Acoustical Society of America held the first in a series of popular sessions devoted to the topic of shock waves in medicine. The goal of the inaugural session was to improve the fundamental understanding of lithotripsy — to bring better devices and treatments to patients. The goal of this paper is to report on progress in this effort

Porcine Models of Pancreatic Cancer

Pancreatic cancer is the fourth most common cause of cancer-related deaths in both men and women. The 5-year survival rate for metastatic pancreatic cancer is only 8%. There remains a need for improved early diagnosis and therapy for pancreatic cancer. Murine models are the current standard for preclinical study of pancreatic cancer. However, mice may not accurately reflect human biology because of a variety of differences between the two species. Remarkably, only 5–8% of anti-cancer drugs that have emerged from preclinical studies and entered clinical studies have ultimately been approved for clinical use. The cause of this poor approval rate is multi-factorial, but may in part be due to use of murine models that have limited accuracy with respect to human disease. Murine models also have limited utility in the development of diagnostic or interventional technology that require a human-sized model. So, at present, there remains a need for improved animal models of pancreatic cancer. The rationale for a porcine model of pancreatic cancer is (i) to enable development of diagnostic/therapeutic devices for which murine models have limited utility; and (ii) to have a highly predictive preclinical model in which anti-cancer therapies can be tested and optimized prior to a clinical trial. Recently, pancreatic tumors were induced in transgenic Oncopigs and porcine pancreatic ductal cells were transformed that contain oncogenic KRAS and p53-null mutations. Both techniques to induce pancreatic tumors in pigs are undergoing further refinement and expansion. The Oncopig currently is commercially available, and it is conceivable that other porcine models of pancreatic cancer may be available for general use in the near future