Investigating in-field and out-of-field neutron contamination in high-energy medical linear accelerators based on the treatment factors of field size,depth, beam modifiers,and beam type

PurposeWe analysed the effects of field size, depth, beam modifier and beam type on the amount of in-field and out-of-field neutron contamination for medical linear accelerators (linacs).MethodsMeasurements were carried out for three high-energy medical linacs of Elekta Synergy Platform, Varian Clinac DHX High Performance and Philips SL25 using bubble detectors. The photo-neutron measurements were taken in the first two linacs with 18 MV nominal energy, whereas the electro-neutrons were measured in the three linacs with 9 MeV, 10 MeV, 15 MeV and 18 MeV.ResultsThe central neutron doses increased with larger field sizes as a dramatic drop off was observed in peripheral areas. Comparing with the jaws-shaped open-field of 10 × 10 cm, the motorised and physical wedges contributed to neutron contamination at central axis by 60% and 18%, respectively. The similar dose increment was observed in MLC-shaped fields. The contributions of MLCs were in the range of 55–59% and 19–22% in Elekta and Varian linacs comparing with 10 × 10 and 20 × 20 cm open fields shaped by the jaws, respectively. The neutron doses at shallow depths were found to be higher than the doses found at deeper regions. The electro-neutron dose at the 18 MeV energy was higher than the doses at the electron energies of 15 MeV and 9 MeV by a factor of 3 and 50, respectively.ConclusionThe photo- and electro-neutron dose should be taken into consideration in the radiation treatment with high photon and electron energies.     

Fibroblast growth factor-2 induces the activation of Src through Fes, which regulates focal adhesion disassembly

Cell migration is regulated by focal adhesion (FA) turnover. Fibroblast growth factor-2 (FGF-2) induces FA disassembly in the murine brain capillary endothelial cell line IBE, leading to FGF-2-directed chemotaxis. We previously showed that activation of Src and Fes by FGF-2 was involved in chemotaxis of IBE cells. In this study, we examined the interplay between Src and Fes. FGF-2 treatment decreased the number of FA in IBE cells, but not in cells expressing dominant-negative Fes (denoted KE5-15 cells). FGF-2 induced the activation of Src and subsequent binding to and phosphorylation of Cas in IBE cells, but not in KE5-15 cells. Focal adhesion kinase (FAK) activation and tyrosine phosphorylation by Src were also delayed in KE5-15 cells compared to parental cells. FGF-2 induced activation of Src within FA in IBE cells, but not in KE5-15 cells. Downregulation of Fes or FAK using small interfering RNA diminished Src activation by FGF-2 within FA. These findings suggest that activation of Fes by FGF-2 enhances FAK-dependent activation of Src within FA, promoting FGF-2-induced disassembly of focal adhesions.     

A novel fluorescent method employing the FRET-based biosensor “LIBRA” for the identification of ligands of the inositol 1,4,5-trisphosphate receptors

LIBRA is a fluorescent biosensor of inositol 1,4,5-trisphosphate (IP₃) and is composed of the ligand-binding domain of the rat type 3 IP₃ receptor and cyan and yellow fluorescent proteins. We examined the responses of LIBRA and its IP₃-insensitive mutant LIBRA-N to compounds known to inhibit IP₃-induced Ca²⁺ release. Heparin, a competitive antagonist of IP₃ receptors, increased the emission ratio of LIBRA but not that of LIBRA-N. In contrast, 2-aminoethoxydiphenyl borate, a known non-competitive inhibitor of IP₃ receptor, decreased the emission ratios of both LIBRA and LIBRA-N. Thus, the concurrent use of LIBRA-N with LIBRA identifies nonspecific responses. These results indicate that LIBRA and its mutant control can be used to detect specific agonists and antagonists of IP₃ receptors. We also demonstrate the utility of LIBRA and LIBRA-N in discriminating between specific and nonspecific responses in intact cells.     

Temporal integrity of an airborne odor stimulus is greatly affected by physical aspects of the odor delivery system

There is currently a debate about the role played by temporal patterns in neural activity in olfactory coding. An accurate analysis of this question, however, is only possible if the temporal properties of a stimulus itself are well defined. So far, no technique with sufficient temporal resolution has been available to accomplish this. Using a photoionization detector (PID), we show that the configuration of the odor delivery apparatus and the airflow settings greatly influence the integrity of a stimulus profile within an odor delivery apparatus. In a situation where pulsatile odor stimuli are applied to a stationary preparation, we tested the effect of 1) axial and off-center location within the airstream, 2) airflow of the odor delivery, 3) exit tube length, 4) exit tube diameter, 5) orientation of the odor delivery device in relation to the exhaust flow, and 6) exhaust tube air speed. This has important implications for the study of time in olfaction; significant planning must be incorporated into the design of the experiment to provide a well-defined odor delivery system.     

Solvent Friction Changes the Folding Pathway of the Tryptophan Zipper TZ2

Because the rate of a diffusional process such as protein folding is controlled by friction encountered along the reaction pathway, the speed of folding is readily tunable through adjustment of solvent viscosity. The precise relationship between solvent viscosity and the rate of diffusion is complex and even conformation-dependent, however, because both solvent friction and protein internal friction contribute to the total reaction friction. The heterogeneity of the reaction friction along the folding pathway may have subtle consequences. For proteins that fold on a multidimensional free-energy surface, an increase in solvent friction may drive a qualitative change in folding trajectory. Our time-resolved experiments on the rapidly and heterogeneously folding β-hairpin TZ2 show a shift in the folding pathway as viscosity increases, even though the energetics of folding is unaltered. We also observe a nonlinear or saturating behavior of the folding relaxation time with rising solvent viscosity, potentially an experimental signature of the shifting pathway for unfolding. Our results show that manipulations of solvent viscosity in folding experiments and simulations may have subtle and unexpected consequences on the folding dynamics being studied.     

Structural and Functional Studies of the Yeast Class II Hda1 Histone Deacetylase Complex

Yeast class II Hda1 histone deacetylase (HDAC) complex is an H2B- and H3-specific HDAC in Saccharomyces cerevisiae consisting of three previously identified subunits, the catalytic subunit scHda1p and two non-catalytic structural subunits scHda2p and scHda3p. We co-expressed and co-purified recombinant yeast class II HDAC complex from bacteria as a functionally active and trichostatin-A-sensitive hetero-tetrameric complex. According to an extensive analysis of domain organization and interaction of all subunits (or domains), the N-terminal domain of scHda1p associates through the C-terminal coiled-coil domains (CCDs) of the scHda2p-scHda3p sub-complex, yielding a truncated scHda1pHDAC-scHda2pCCD2-scHda3pCCD3 complex with indistinguishable deacetylase activity compared to the full-length complex in vitro. We characterized the interaction of the HDAC complex with either single-stranded or double-stranded DNA and identified the N-terminal halves of scHda2p and scHda3p as binding modules. A high-resolution structure of the scHda3p DNA-binding domain by X-ray crystallography is presented. The crystal structure shows an unanticipated structural homology with the C-terminal helicase lobes of SWI2/SNF2 chromatin-remodeling domains of the Rad54 family enzymes. DNA binding is unspecific for nucleotide sequence and structure, similar to the Rad54 enzymes in vitro. Our structural and functional analyses of the budding yeast class II Hda1 HDAC complex provide insight into DNA recognition and deacetylation of histones in nucleosomes.     

Ryanodine receptor mutations in arrhythmia: The continuing mystery of channel dysfunction

Mutations in RyR2 are causative of an inherited disorder which often results in sudden cardiac death. Dysfunctional channel behaviour has been the subject of many investigations varying from single channel analysis through to complex animal models. This review discusses recent advances in the field, describes the controversy surrounding the exact consequences of RyR2 mutation and how the disparate data may be reconciled. This heterogeneity of function with respect to the effects of polymorphisms, phosphorylation, cytosolic and luminal Ca²⁺ as well as inter-domain interactions may have important implications for the recent pharmaceutical therapies which have been put forward. We surmise that a comprehensive characterisation of mutations on a case-by-case basis may be beneficial for the development of specifically targeted therapies.     

Detectability of American Mink Using Rafts to Solicit Field Signs in a Population Control Context

Abstract: American mink (Neovison vison) are an ecologically damaging invasive species where they have been introduced in Europe. Effectiveness of mink population control by trapping has been difficult to assess, without knowing how efficiently mink are caught by traps or detected by other methods. Use of track-recording rafts to detect mink and guide trapping effort has proved efficient and leads to a supposition that no detection indicates absence of mink. To draw this conclusion with any confidence requires a measure of detectability. We applied occupancy models to data from an earlier study to estimate detectability of individual American mink on track-recording rafts. Estimated detectability of individual mink, per raft, and 2-week check period varied between 0.4 in late summer and 0.6 in late autumn. By inference, risk of failing to detect a mink that was present would be <5% given 4–6 independent opportunities to detect it. These opportunities could be created either by using a raft spacing that ensured multiple detections of each mink or by monitoring rafts through a succession of check intervals. Within certain simple constraints, raft location did not contribute substantially to detection probability. These findings will allow field operators, strategists, and funders to assess with confidence the success of efforts to control mink density. We expect the estimation of individual detectability to be similarly valuable in population control or eradication of other species.     

Gold finder: a computer method for fast automatic double gold labeling detection,counting, and color overlay in electron microscopic images

This work presents a computerized method to identify, detect, evaluate, and, by colored overlay, present gold particle pairs in electron microscopy (EM), even in wide-field views. Double gold immunolabeled specimens were analyzed in a LEO 912 electron microscope equipped with a 2k x 2k-pixel slow-scan cooled CCD camera connected to a computer with analySIS 3.1 PRO image processing software. The acquisition of a high-resolution and high-dynamic-range image by the camera allowed correct segmentation of the gold particles, separating them from other cell structures and from the substrate. Particle identification was performed by a classification module designed by us. Based on shape and size, the computer recognized the group of small particles and classified them as either singular or clustered and differentiated these from the single bigger type. The final image shows the particle types separated and colored, and indicates the total number of objects encountered in the specific region of interest. Moreover, a montage tool allowed us to obtain final representative images of large microscopic fields, which on analysis by the Gold Finder module provided information on the distribution and localization of antigens comparable to that provided by the wide-field light microscope images.     

Hydrolyzable ATP and PIP(2) modulate the small-conductance K+ channel in apical membranes of rat cortical-collecting duct (CCD)

The small-conductance K+ channel (SK) in the apical membrane of the cortical-collecting duct (CCD) is regulated by adenosine triphosphate (ATP) and phosphorylation-dephosphorylation processes. When expressed in Xenopus oocytes, ROMK, a cloned K+ channel similar to the native SK channel, can be stimulated by phosphatidylinositol bisphosphate (PIP2), which is produced by phosphoinositide kinases from phosphatidylinositol. However, the effects of PIP2 on SK channel activity are not known. In the present study, we investigated the mechanism by which hydrolyzable ATP prevented run-down of SK channel activity in excised apical patches of principal cells from rat CCD. Channel run-down was significantly delayed by pretreatment with hydrolyzable Mg-ATP, but ATP gamma S and AMP-PNP had no effect. Addition of alkaline phosphatase also resulted in loss of channel activity. After run-down, SK channel activity rapidly increased upon addition of PIP2. Exposure of inside-out patches to phosphoinositide kinase inhibitors (LY294002, quercetin or wortmannin) decreased channel activity by 74% in the presence of Mg-ATP. PIP2 added to excised patches reactivated SK channels in the presence of these phosphoinositide kinase inhibitors. The protein kinase A inhibitor, PKI, reduced channel activity by 36% in the presence of Mg-ATP. PIP2 was also shown to modulate the inhibitory effects of extracellular and cytosolic ATP. We conclude that both ATP-dependent formation of PIP2 through membrane-bound phosphoinositide kinases and phosphorylation of SK by PKA play important roles in modulating SK channel activity.