First clinical experience with real-time portal imaging-based breath-hold monitoring in tangential breast radiotherapy.

Background and purpose: Real-time treatment monitoring with the electronic portal imaging device (EPID) can conceptually provide a more accurate assessment of the quality of deep inspiration breath-hold (DIBH) and patient movement during tangential breast radiotherapy (RT). A system was developed to measure two geometrical parameters, the lung depth (LD) and the irradiated width (named here skin distance, SD), along three user-selected lines in MV EPID images of breast tangents. The purpose of this study was to test the system during tangential breast RT with DIBH. Materials and methods: Measurements of LDs and SDs were carried out in real time. DIBH was guided with a commercial system using a marker block. Results from 17 patients were assessed. Mean midline LDs, , per tangent were compared to the planned mLDs; differences between the largest and smallest observed () per tangent were calculated. Results: For 56% (162/288) of the tangents tested, were outside the tolerance window. All but one patient had at least one fraction showing this behaviour. The largest difference found between an and its planned mLD was -16.9 mm. The accuracy of patient positioning and the quality of marker-block-based DIBH guidance contributed to the differences. Fractions with patient position verification using a single EPID image taken before treatment showed a lower rate (34%), suggesting reassessment of setup procedures. Conclusions: Real-time treatment monitoring of the internal anatomy during DIBH delivery of tangential breast RT is feasible and useful. The new system requires no additional radiation for the patient.

A system for real-time monitoring of breath-hold via assessment of internal anatomy in tangential breast radiotherapy.

The deep inspiration breath-hold (DIBH) technique assists in sparing the heart, lungs, and liver during breast radiotherapy (RT). The quality of DIBH is currently assessed via surrogates which correlate to varying degrees with the patient's internal anatomy. Since modern linacs are equipped with an electronic portal imaging device (EPID), images of the irradiated anatomy streamed from EPIDs and analyzed in real time could significantly improve assessment of the quality of DIBH. A system has been developed to quantify the quality of DIBH during tangential breast RT by analyzing the "beam's eye view" images of the treatment fields. The system measures the lung depth (LD) and the distance from the breast surface to the posterior tangential radiation field edge (skin distance, SD) at three user-defined locations. LD and SD measured in real time in EPID images of two RT phantoms showing different geometrical characteristics of their chest wall regions (computed tomography dose index [CTDI] and "END-TO-END" stereotactic body radiation therapy [E2E SBRT]) were compared with ground truth displacements provided by a precision motion platform. Performance of the new system was evaluated via static and dynamic (sine wave motion) measurements of LD and SD, covering clinical situations with stable and unstable breath-hold. The accuracy and precision of the system were calculated as the mean and standard deviation of the differences between the ground truth and measured values. The accuracy of the static measurements of LD and SD for the CTDI phantom was 0.31 (1.09) mm [mean (standard deviation)] and -0.10 (0.14) mm, respectively. The accuracy of the static measurements for E2E SBRT phantom was 0.01 (0.18) mm and 0.05 (0.08) mm. The accuracy of the dynamic LD and SD measurements for the CTDI phantom was -0.50 (1.18) mm and 0.01 (0.12) mm, respectively. The accuracy of the dynamic measurements for E2E SBRT phantom was -0.03 (0.19) mm and 0.01 (0.11) mm.

Multilayer Capsules of Bovine Serum Albumin and Tannic Acid for Controlled Release by Enzymatic Degradation.

With the purpose to replace expensive and significantly cytotoxic positively charged polypeptides in biodegradable capsules formed via Layer-by-Layer (LbL) assembly, multilayers of bovine serum albumin (BSA) and tannic acid (TA) are obtained and employed for encapsulation and release of model drugs with different solubility in water: hydrophilic-tetramethylrhodamine-isothiocyanate-labeled BSA (TRITC-BSA) and hydrophobic 3,4,9,10-tetra-(hectoxy-carbonyl)-perylene (THCP). Hydrogen bonding is proposed to be predominant within thus formed BSA/TA films. The TRITC-BSA-loaded capsules comprising 6 bilayers of the protein and polyphenol are benchmarked against the shells composed of dextran sulfate (DS) and poly-l-arginine (PARG) on degradability by two proteolytic enzymes with different cleavage site specificity (i.e., α-chymotrypsin and trypsin) and toxicity for murine RAW264.7 macrophage cells. Capsules of both types possess low cytotoxicity taken at concentrations equal or below 50 capsules per cell, and evident susceptibility to α-chymotrypsin resulted in release of TRITC-BSA. While the BSA/TA-based capsules clearly display resistance to treatment with trypsin, the assemblies of DS/PARG extensively degrade. Successful encapsulation of THCP in the TRITC-BSA/TA/BSA multilayer is confirmed, and the release of the model drug is observed in response to treatment with α-chymotrypsin. The thickness, surface morphology, and enzyme-catalyzed degradation process of the BSA/TA-based films are investigated on a planar multilayer comprising 40 bilayers of the protein and polyphenol deposited on a silicon wafer. The developed BSA/TA-based capsules with a protease-specific degradation mechanism are proposed to find applications in personal care, pharmacology, and the development of drug delivery systems including those intravenous injectable and having site-specific release capability.

Avidin adsorption on silicone elastomer: effects of stretching and polyelectrolyte coatings.

Zeta potential and avidin adsorption of silicone elastomer (SE) and SE coated with polyethyleneimine (PEI) and heparin is studied under controlled hydrodynamic conditions. Zeta potential (ζ) of SE depends on the duration of contact with buffer (0.02M MOPS-KCl, pH 7.2): ζ descends from 12mV to a plateau value of -32mV during 5h and decreases further if SE is stretched (30% elongation). The time variation of ζ is attributed to surface charging of the core polymer PDMS. Coating the negatively charged SE with PEI results in ζ=42mV; a layer of heparin on top of PEI yields ζ=-46mV. On the negatively charged surfaces, avidin adsorption levels off at 5mV. Desorption of avidin from SE is negligible even when SE with adsorbed avidin is stretched, or when the stretching force is removed after adsorption of avidin. The results can be employed for designing biomaterials with properties such as surface charge and protein adsorption adjustable by stretching; the strong attachment of avidin can allow for further surface modification.

The BAD project: data mining, database and prediction of protein adsorption on surfaces.

Protein adsorption at solid-liquid interfaces is critical to many applications, including biomaterials, protein microarrays and lab-on-a-chip devices. Despite this general interest, and a large amount of research in the last half a century, protein adsorption cannot be predicted with an engineering level, design-orientated accuracy. Here we describe a Biomolecular Adsorption Database (BAD), freely available online, which archives the published protein adsorption data. Piecewise linear regression with breakpoint applied to the data in the BAD suggests that the input variables to protein adsorption, i.e., protein concentration in solution; protein descriptors derived from primary structure (number of residues, global protein hydrophobicity and range of amino acid hydrophobicity, isoelectric point); surface descriptors (contact angle); and fluid environment descriptors (pH, ionic strength), correlate well with the output variable-the protein concentration on the surface. Furthermore, neural network analysis revealed that the size of the BAD makes it sufficiently representative, with a neural network-based predictive error of 5% or less. Interestingly, a consistently better fit is obtained if the BAD is divided in two separate sub-sets representing protein adsorption on hydrophilic and hydrophobic surfaces, respectively. Based on these findings, selected entries from the BAD have been used to construct neural network-based estimation routines, which predict the amount of adsorbed protein, the thickness of the adsorbed layer and the surface tension of the protein-covered surface. While the BAD is of general interest, the prediction of the thickness and the surface tension of the protein-covered layers are of particular relevance to the design of microfluidics devices.

Fate of prions in soil: adsorption kinetics of recombinant unglycosylated ovine prion protein onto mica in laminar flow conditions and subsequent desorption.

Prions can be disseminated in soils. Their interaction with soil minerals is a key factor for the assessment of risks associated with the transport of their infectivity. We did not examine here the infectivity itself but the adsorption kinetics of an ovine recombinant prion protein (ovine PrPrec), as a noninfectious model protein, on muscovite mica, a phyllosilicate with surface properties analogous to soil clays, in conditions of laminar flow through a channel. The protein was labeled with (125)I, and its adsorption examined between pH 4.0 and 9.0. At wall shear rate 100 s(-1), we found the process to be controlled mainly by transport at the beginning of the adsorption process. Additional experiments at 1000 s(-1) (pH 5 and 6) determined that the diffusion coefficient was in accordance with the hydrodynamic radius measured by size exclusion chromatography. The pseudo-plateau of the interfacial concentration with time was compatible with more than a monolayer and suggests the presence of aggregates. Desorption was not observed in the presence of buffer between pH 4 and 9 and sheep plasma, while the addition of alkaline detergent or 10(-1) M NaOH allowed an almost complete removal from the interface. The ensemble of results suggests that the largely irreversible adsorption of the ovine PrPrec onto mica is mainly due to electrostatic attraction between the protein and the highly negatively charged mica surface. Possible consequences for the mode of dissemination of prion proteins in soils are indicated.

Streaming potential in cylindrical pores of poly(ethylene terephthalate) track-etched membranes: variation of apparent zeta potential with pore radius.

Streaming potential variation with pressure measured through poly(ethylene terephthalate) track-etched membranes of different pore sizes led to the determination of an apparent interfacial potential zetaa in the presence of 10-2 M KCl. The variation of zetaa with the pore radius r0 is interpreted by (i) the electric double layer overlap effect and (ii) the presence of a conductive gel layer. We propose a method which integrates both effects by assuming a simple model for the conductive gel at the pore wall. We observed the following three domains of pore size: (i) r0 > 70 nm, where surface effects are negligible; (ii) approximately 17 nm < r0 < 70 nm, where the pore/solution interface could be described as a conductive gel of thickness around 1 nm; (iii) r0 < approximately 17 nm, which corresponds to the region strongly damaged by the ion beam and is not analyzed here. The first one (zeta = -36.2 mV) corresponds to the raw material when etching has completely removed the ion beam predamaged region, which corresponds to the second intermediate domain (zeta = -47.3 mV). There the conductance of the gel layer deduced from the treatment of streaming potential data was found to be compatible with the number of ionic sites independently determined by the electron spin resonance technique.

Adsorption of alpha-chymotrypsin onto mica in laminar flow conditions. Adsorption kinetic constant as a function of tris buffer concentration at pH 8.6.

We examined the adsorption kinetics of alpha-chymotrypsin (pH 8.6, 10(-2) to 0.5 M Tris buffer) on muscovite mica in conditions of laminar flow through a slit. The range of buffer concentrations is between two limits: (i) no adsorption in 1 M Tris and (ii) no desorption in 10(-3) M Tris. Studying the dependence of adsorption kinetics on the wall shear rate leads to the determination of the interfacial adsorption kinetic constant ka and the diffusion coefficient. The obtained value for the diffusion coefficient is close to the one expected from the molecular size of alpha-chymotrypsin. The interfacial adsorption kinetic constant of alpha-chymotrypsin decreases when ionic strength increases, while the initial desorption constant (over a part of all the adsorbed population) shows the contrary. Although alpha-chymotrypsin is almost at its isoelectric point, the effect of ionic strength on the adsorption kinetics suggests the importance of electrostatic interactions between the protein and mica. We observed an increase in the adsorption rate, at a surface coverage near 0.14 microg cm(-2), for adsorption in 10(-2) M Tris and the low wall shear rates (<300 s(-1)). This change in the adsorption rate suggests a structural transition, that we assume again to be due to electrostatic interactions, but between proteins. The large dipole moment of the protein may induce this transition, illustrated here by the ferroelectric/antiferroelectric pattern. The variation of the zeta potential with interfacial concentration seems to be in agreement with such a model.

Kinetics of adsorption, desorption, and exchange of alpha-chymotrypsin and lysozyme on poly(ethyleneterephthalate) tracked film and track-etched membrane.

Adsorption kinetics of (125)I-radiolabeled alpha-chymotrypsin at pH 8.6 was studied in a laminar regime between two walls of poly(ethyleneterephthalate) tracked films and membranes. Adsorption kinetics in the presence of solution (10 microg/mL), desorption by rinsing with buffer, and the following exchange of proteins by flowing unlabeled solution were measured. At pH 8.6, alpha-chymotrypsin is almost neutral and can be mostly removed from the film surface, contrary to positive lysozyme adsorbed at pH 7.4. Results suggest that alpha-chymotrypsin is irreversibly adsorbed in pores, while desorption and exchange occur on membrane flat faces. A method is proposed to determine adsorption kinetics in the pores. Kinetics of desorption and exchange of alpha-chymotrypsin from the film surface can be described by stretched exponential functions in the examined time domain with the same exponent, beta approximately 0.62, which does not depend also on the former adsorption duration. However, the mean residence time at the interface is about 2.5 times greater in the presence of only the buffer than that in the presence of solution. This effect could be explained by a fast exchange at the arrival of unlabeled solution for a part of the adsorbed population.