Dosimetric comparisons of helical tomotherapy and step-and-shoot intensity-modulated radiotherapy in nasopharyngeal carcinoma.

PURPOSE: The study evaluates and quantifies the potential dosimetric gains of helical tomotherapy (HT) versus step-and-shoot intensity-modulated radiotherapy (SaS-IMRT) for nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: Twenty consecutive NPC patients curatively treated by HT were examined. Each case was planned by HT and SaS-IMRT (ADAC Pinnacle(3)) planning system, respectively. Dose plans were compared using dose volume histograms (DVH), conformity index (CI), homogeneity index (HI), and minimal dose to 1cc (D(min_1cc)) of the planned target volume (PTV) and a comprehensive quality index (CQI) of ten organs at risk (OARs). The prescribed dose/fractionation was 72Gy to the PTV, 64.8Gy to the elective PTV, and 54Gy to the clinically negative neck region. The plan of 54Gy to the PTV (PTV(54)) was used to evaluate the CI and HI in the target. The cumulative doses of the three PTV plans to the OARs were calculated. RESULTS: We observed the HT plans significantly improved the CI (improvement ratio: 11.9+/-5.5%) and HI (improvement ratio: 8.8+/-1.5%) of the PTV(54) compared with SaS-IMRT plans. In addition, the mean/maximal dose of most of the OARs except chiasm was significantly reduced in HT plans, with the CQI of 0.92+/-0.08. A negative result of HT in chiasm was observed but only significantly revealed in cases without skull base infiltration. CONCLUSIONS: A dosimetric gain in CI and HI of PTV and sparing of OARs was significantly obtained in HT versus SaS-IMRT plans in NPC patients. Whether such dosimetric superiority in HT could transfer into clinical advantages needs further investigation.

Time-resolved FRET and FLIM of four-way DNA junctions.

Conformational transitions in a 4-way DNA junction when titrated with ionic solutions are studied using time-resolved fluorescence resonance energy transfer. Parameters characterising the transition in terms of critical ion concentration (c1/2) and the Hill coefficient for ion binding are obtained by fitting a simple two-state model using steady-state spectra. Data obtained from a fluorescence lifetime plate reader and analysed by fitting a single exponential to donor fluorescence lifetime decays are shown to be in good agreement with the parameters obtained from steady-state measurements. Fluorescence lifetimes, however, offer advantages, particularly in being independent of fluorophore concentration, output intensity, inhomogeneity in the excitation source and output wavelength. We demonstrate preliminary FRET-FLIM images of DNA junction solutions obtained using a picosecond gated CCD which are in agreement with results from a fluorescence lifetime plate reader. The results suggest that time-resolved FRET-FLIM is sensitive to subtle structural changes and may be useful in assays based on 4-way DNA junctions.

The stability and characteristics of a DNA Holliday junction switch.

A Holliday junction (HJ) consists of four DNA double helices, with a branch point discontinuity at the intersection of the component strands. At low ionic strength, the HJ adopts an open conformation, with four widely spaced arms, primarily due to strong electrostatic repulsion between the phosphate groups on the backbones. At high ionic strength, screening of this repulsion induces a switch to a more compact (closed) junction conformation. Fluorescent labelling with dyes placed on the HJ arms allows this conformational switch to be detected optically using fluorescence resonance energy transfer (FRET), producing a sensitive fluorescent output of the switch state. This paper presents a systematic and quantitative survey of the switch characteristics of such a labelled HJ. A short HJ (arm length 8 bp) is shown to be prone to dissociation at low switching ion concentration, whereas an HJ of arm length 12 bp is shown to be stable over all switching ion concentrations studied. The switching characteristics of this HJ have been systematically and quantitatively studied for a variety of switching ions, by measuring the required ion concentration, the sharpness of the switching transition and the fluorescent output intensity of the open and closed states. This stable HJ is shown to have favourable switch characteristics for a number of inorganic switching ions, making it a promising candidate for use in nanoscale biomolecular switch devices.

Surface adsorption of zwitterionic surfactants: n-alkyl phosphocholines characterised by surface tensiometry and neutron reflection.

The surface adsorption of n-dodecyl phosphocholine (C12PC) has been characterised by a combined measurement of surface tension and neutron reflectivity. The critical micellar concentration (CMC) was found to be 0.91 mM at 25 degrees C in pure water. At the CMC, the limiting area per molecule (A(cmc)) was found to be 52+/-3 A2 and the surface tension (gamma(cmc)) to be ca. 40.0+/-0.5 mN/m. The parallel study of chain isomer n-hexadecyl phosphocholine (C16PC) showed a decrease of the CMC to 0.012 mM and a drop of gamma(cmc) to 38.1+/-0.5 mN/m. However, A(cmc) for C16PC was found to be 54+/-3 A2, showing that increase in alkyl chain length by four methylene groups has little effect on A(cmc). The almost constant A(cmc) suggested that the limiting area per molecule was determined by the bulky PC head group. It was further found that the surface tension and related key physical parameters did not vary much with temperature, salt addition, solution pH or any combination of these, thus showing that surface adsorption and solution aggregation from PC surfactants is largely similar to the zwitterionic betaine surfactants and is distinctly different from ionic and non-ionic surfactants. The thickness of the adsorbed monolayers measured from both dC12hPC and dC16hPC was found to be 20-22 A at the CMC from neutron reflectivity. Neither A(cmc) nor layer thickness varied with alkyl chain length, indicating that as the alkyl chain length became longer it was further tilted away from the surface normal direction and the layer packing density increased. It was also observed that the thickness of the layer varied little with surfactant concentration, indicating that the average conformational orientation of the alkyl chain remained unchanged against varying surface coverage.

Single molecule fluorescence imaging and its application to the study of DNA condensation.

Single molecule fluorescence imaging incorporated with optical tweezers and a laminar flow cell has been used to monitor the kinetic process of DNA condensation induced by spermidine. It was found that at least two steps were involved in the condensation process of the hydrodynamically-stretched linear DNA; a lag period followed by a rapid collapse of DNA. The lag time increased with the flow speed and the collapse time remained short within the range of the flow speed studied. The effect of salt concentration on the condensation process was examined, and the results suggest that the longer lag time observed in the higher salt buffer probably results from the displacement of bound cations and rearrangement of spermidine on the DNA. The flow-speed dependence of the lag time suggests that a nucleation event at the free end of the DNA, i.e. formation of a loop, may play a vital role in the kinetic process of condensation.

Unusual 2-aminopurine fluorescence from a complex of DNA and the EcoKI methyltransferase.

The methyltransferase, M.EcoKI, recognizes the DNA sequence 5'-AACNNNNNNGTGC-3' and methylates adenine at the underlined positions. DNA methylation has been shown by crystallography to occur via a base flipping mechanism and is believed to be a general mechanism for all methyltransferases. If no structure is available, the fluorescence of 2-aminopurine is often used as a signal for base flipping as it shows enhanced fluorescence when its environment is perturbed. We find that 2-aminopurine gives enhanced fluorescence emission not only when it is placed at the M.EcoKI methylation sites but also at a location adjacent to the target adenine. Thus it appears that 2-aminopurine fluorescence intensity is not a clear indicator of base flipping but is a more general measure of DNA distortion. Upon addition of the cofactor S-adenosyl-methionine to the M.EcoKI:DNA complex, the 2-aminopurine fluorescence changes to that of a new species showing excitation at 345 nm and emission at 450 nm. This change requires a fully active enzyme, the correct cofactor and the 2-aminopurine located at the methylation site. However, the new fluorescent species is not a covalently modified form of 2-aminopurine and we suggest that it represents a hitherto undetected physicochemical form of 2-aminopurine.

Interaction of the ocr gene 0.3 protein of bacteriophage T7 with EcoKI restriction/modification enzyme.

The ocr protein, the product of gene 0.3 of bacteriophage T7, is a structural mimic of the phosphate backbone of B-form DNA. In total it mimics 22 phosphate groups over approximately 24 bp of DNA. This mimicry allows it to block DNA binding by type I DNA restriction enzymes and to inhibit these enzymes. We have determined that multiple ocr dimers can bind stoichiometrically to the archetypal type I enzyme, EcoKI. One dimer binds to the core methyltransferase and two to the complete bifunctional restriction and modification enzyme. Ocr can also bind to the component subunits of EcoKI. Binding affinity to the methyltransferase core is extremely strong with a large favourable enthalpy change and an unfavourable entropy change. This strong interaction prevents the dissociation of the methyltransferase which occurs upon dilution of the enzyme. This stabilisation arises because the interaction appears to involve virtually the entire surface area of ocr and leads to the enzyme completely wrapping around ocr.

Inhibition of toxicity and protofibril formation in the amyloid-beta peptide beta(25-35) using N-methylated derivatives.

Beta (25-35) is a fragment of beta-amyloid that retains its wild-type properties. N-methylated derivatives of beta(25-35) can block hydrogen bonding on the outer edge of the assembling amyloid, so preventing the aggregation and inhibiting the toxicity of the wild-type peptide. The effects are assayed by Congo Red and thioflavin T binding, electron microscopy and an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] toxicity assay. N-methyl-Gly-25 has similar properties to the wild- type, while five other methylation sites have varying effects on prefolded fibrils and fibril assembly. In particular, N-methyl-Gly-33 is able to completely prevent fibril assembly and reduces the toxicity of prefolded amyloid. With N-methyl-Leu-34 the fibril morphology is altered and toxicity reduced. A preliminary study of beta(25-35) structure in aqueous solution was made by small-angle neutron scattering (SANS). The protofibrillar aggregates are best described as a disc of radius 140 A and height 53 A (1 A = 0.1 nm), though the possibility of polydisperse aggregates cannot be ruled out. No aggregates form in the presence of N-methyl-Gly-33. We suggest that the use of N-methylated derivatives of amyloidogenic peptides and proteins could provide a general solution to the problem of amyloid deposition and toxicity and that SANS is an important technique for the direct observation of protofibril formation and destruction in solution.

Beta-casein adsorption at the hydrophobized silicon oxide-aqueous solution interface and the effect of added electrolyte.

The effect of the presence of NaCl, CaCl(2), or MgCl(2) at the same ionic strength on the structure of beta-casein layers adsorbed on hydrophobic surfaces has been investigated by neutron reflectivity measurements. The data were fitted to a four-layer model. The volume fraction versus distance profiles have a similar shape whether beta-casein is adsorbed from NaCl, CaCl(2), and MgCl(2) of the same ionic strength or whether the protein concentration is lowered 10 times. In particular at larger distances from the surface, the volume fraction values are low and similar. However, close to the hydrophobic surface the volume fraction of protein decreases in the order CaCl(2) > MgCl(2) > NaCl. We have also used a specific proteolytic enzyme, endoproteinase Asp-N, which cleaves off the hydrophilic part of beta-casein, as a tool to reveal the interfacial structure of the protein. For all the different types of added electrolytes, endoproteinase Asp N only affects the outermost beta-casein layer. Subsequent addition of beta-casein in all cases led to large increases in amounts adsorbed and in the thickness of the outer layers.

beta-Casein adsorption at the silicon oxide--aqueous solution interface.

Neutron reflectometry was used to investigate the time-dependent beta-casein adsorption at the silica-aqueous solution interface. The transient and steady-state structural characteristics of the adsorbed layer were determined from reflectivity curves, fitted to three-layer and two-layer models. The results show that the beta-casein adsorption to silica is very slow. The adsorption process involves the formation of an inner dense protein layer with a mean thickness of about 30 A onto which a more hydrated outer layer is self-associated. The surface excess and the total layer thickness of the asymmetric bilayer were, after 5 h adsorption time, estimated to be about 6.5 mg/m2 and 105 A, respectively. The adsorption behavior observed on silica contrasts with that previously reported for hydrophobic substrates, where beta-casein adsorption is much more rapid and the final surface excess is less than half of that observed for silica. Rinsing the silica surface with protein-free buffer resulted in a substantial desorption; much more pronounced than observed for hydrophobic substrates. This behavior suggests a weak adsorption affinity for a fraction of the adsorbed casein molecules; most likely the outer self-associated casein molecules in the adsorbed bilayer. The comparative desorption from hydrophobic surfaces was shown to be marginal. The difference between the layer structures adopted on hydrophobic and hydrophilic surfaces is also mirrored in the effects that the addition of a specific proteolytic enzyme (endoproteinase Asp-N) has on the adsorbed layer properties. The rinsing and endoproteinase cleavage processes result together in more than 80% reduction of the originally adsorbed mass at the silica surface. Only a thin but dense adsorbed layer remains after these treatments. The corresponding reduction reported for the hydrophobic adsorbent system was only about 20%. It is concluded that beta-casein adsorption on silica results in the formation of an asymmetric surface bound bilayer that stands in strong contrast to the monolayer structure formed at hydrophobic surfaces. This finding support the previous results obtained by using ellipsometry. The study also shows that neutron reflection, despite its limitations in time resolution, can be used for studying dynamic interfacial phenomena in protein systems.

Comparative study of daily activities of pregnant and non-pregnant women after in vitro fertilization and embryo transfer.

BACKGROUND AND PURPOSE: The purposes of this study were to explore daily activity following embryo transfer (ET) in women who had undergone in vitro fertilization (IVF), and to compare the differences between pregnant and non-pregnant cycles. METHODS: This prospective survey was conducted in a medical center in northern Taiwan. Subjects were women who had completed one IVF cycle. Using a structured questionnaire, the subjects were asked whether they had changed their normal daily activities during the 2-week waiting period following ET. Data were collected on the day of ET and on the day of pregnancy testing before the outcome of the treatment was known. After the results of the pregnancy test were known, subjects were divided into pregnant and non-pregnant groups, with 30 subjects in each group. RESULTS: Of the 60 participants, 56 reported that they had tried to rest in bed for more than 2 hours following ET, and 54 reported that they limited their self-care activity on the day of ET. During the 2-week waiting period following ET, 55 walked at a slow pace, 51 limited their social activity, 47 avoided the use of stairs, 45 limited their self-care activity, and 40 adjusted their work load. However, a comparison of daily activity variables between the two groups yielded no significant differences. CONCLUSIONS: Most subjects reported that they restricted their daily activities following IVF/ET, even though their doctors suggested that they return to their normal routines. Bed rest was not correlated with successful IVF.

Neutron reflection from a dimyristoylphosphatidylcholine monolayer adsorbed on a hydrophobised silicon support.

Neutron specular reflection has been used to study the structure of a monolayer of dimyristoylphosphatidylcholine (DMPC) deposited using the Langmuir-Blodgett technique onto a silicon oxide substrate. A self-assembled monolayer of octadecyltrichlorosilane with a deuterated alkyl chain (d-OTS) had been previously bonded onto this silicon oxide substrate which rendered it hydrophobic. In the system under study, the alkyl chains of the phospholipid were found to penetrate extensively into the d-OTS layer with the mixed chain region (d-OTS and DMPC) having a total thickness of 30.5 A. This mixed region was divided into two halves for analysis; the 'lower half' (nearest to the substrate surface) was found to comprise anchored d-OTS chains mixed with the lipid chains in the volume ratio approx. 0.60:0.35. The corresponding volume ratio in the 'upper half' of this region was determined to be approx. 0.50:0.40. The thicknesses of these regions were found to be 17.9 A (incorporating approx. 6% solvent) and 12.6 A (incorporating approx. 9% solvent) for the lower and upper halves respectively. The DMPC head groups were found to be confined to the most external layer (furthest away from the silicon substrate). This layer was found to have a thickness of 9.4 A and included a small fraction of the lipid alkyl chains with approx. 47% solvent.

[The experience and adaptative behaviors of the infertile women who were suffering from in vitro fertilization failure].

The purposes of this study were to explore the women's experience and adaptative behaviors when they suffered from in vitro fertilization failure. Under a survey designed using purposive sampling, 30 IVF fertilization failure women were recrutied. A semi-structured interview guide was developed for data collection. When the subjects were informed for the IVF fertilization failure, they were interviewed and data were collected. All data were translated in a narrative form and analyzed by content analysis. The results showed there were four types of experience and each of them had different adaptative behaviors: (A) They were disappointed and confused to the fertilization failure; they coped by accepting the fate, decreasing the expectation, looking for the failing factors, and looking for the successful factors. (B) They felt painful during the treatment course; they coped by reviewing the treatment course, having a rest, doing something that they had delayed, and planning to go ahead. (C) They felt unfair to the outcome of the effort; they coped by blaming the fate, wanting to change the mode of treatment, or by refusing to accept the treatment again. (D) They did not know what to do next; they coped by avoiding the fact or searching for more consultation. The findings can be used to establish a nursing care standard for the IVF fertilization failure women. Accordingly, the nurses can help the infertile women to recover from their grief and restitute a positive prospect of their future lives.

Planar fabrication process of a high-coupling-efficiency interface between optical waveguides of large index difference.

A high-coupling-efficiency interface connecting two optical waveguides of large index difference is reported. Both theoretical analysis and an experimental study were performed. High coupling efficiency is achieved by matching the mode profiles in two waveguides as well as connecting the waveguides with an antireflection (AR) section. Analysis shows a coupling efficiency as high as 98% between glass-SiO(2) and Ga(0.72)Al(0.28)As-Ga(0.59)Al(0.41)As waveguides with an AR section of TiO(2). An index difference of 1.8 for the TE(0) mode between these two waveguides is obtained. To implement the glass-SiO(2) waveguide with an AR section in the host AlGaAs waveguide, a new quadrilevel photomasking and lift-off process is developed. A coupling efficiency of 83% was measured for the TE(0) mode at the 1.15-µm wavelength, a very high value considering the large index difference (1.75) between the two waveguides that were connected. The design concept and the fabrication process developed should facilitate efficient integration of low- and high-index waveguides on the same substrate to produce a greater variety of photonic devices and modules.

2 × 2 single-mode zero-gap directional-coupler thermo-optic waveguide switch on glass.

A 2 × 2 single-mode thermo-optic waveguide switch fabricated on a glass substrate is reported. The device uses a zero-gap directional-coupler configuration with a heater electrode to increase the waveguide index of refraction by the use of the thermo-optic effect. With an interaction length of 6 mm and using 0.63-µm He-Ne laser light, the device experiences a complete switchover of light from a straight-through channel to a crossed channel at an electrical drive power of 0.25 W under a quasi-steady-state condition and 0.67 W under a pulsed condition of 800 -s in pulse width. The device response is independent of light polarization and insensitive to ambient temperature. Transient thermal analysis is carried out with a two-dimensional transient temperature solution derived previously [IEEE Trans. Components Hybrids Manuf. Technol. 13, 980 (1990)]. A thermo-optic equation is established to relate the optical intensity to the temperature rise in the waveguide. The deduced optical responses based on the thermo-optic equation and the calculated thermal response agree extremely well with the measured responses, thus verifying the thermo-optic equation and the thermal-analysis method. We also confirm the fundamental principle of the zero-gap directional coupler and find that the 2 × 2 single-mode switch will be a useful component in future fiber-optic and integrated-optic systems.

Ascending cholangitis complicated by pyogenic Iiver abscess.

No abstract available.