Generation of recombinant hyperimmune globulins from diverse B-cell repertoires.

Plasma-derived polyclonal antibody therapeutics, such as intravenous immunoglobulin, have multiple drawbacks, including low potency, impurities, insufficient supply and batch-to-batch variation. Here we describe a microfluidics and molecular genomics strategy for capturing diverse mammalian antibody repertoires to create recombinant multivalent hyperimmune globulins. Our method generates of diverse mixtures of thousands of recombinant antibodies, enriched for specificity and activity against therapeutic targets. Each hyperimmune globulin product comprised thousands to tens of thousands of antibodies derived from convalescent or vaccinated human donors or from immunized mice. Using this approach, we generated hyperimmune globulins with potent neutralizing activity against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in under 3 months, Fc-engineered hyperimmune globulins specific for Zika virus that lacked antibody-dependent enhancement of disease, and hyperimmune globulins specific for lung pathogens present in patients with primary immune deficiency. To address the limitations of rabbit-derived anti-thymocyte globulin, we generated a recombinant human version and demonstrated its efficacy in mice against graft-versus-host disease.

Technical Note: Implications of using EGSnrc instead of EGS4 for extracting electron stopping powers from measured energy spectra.

PURPOSE: NRC Report PIRS-0626 (https://doi.org/10.4224/40000364) describes how measured electron energy deposition spectra can be used to determine the electronic stopping power. The stopping power is obtained by comparing measured spectra with spectra calculated using Monte Carlo techniques. The stopping powers reported in PIRS-0626 were obtained using the EGS4 Monte Carlo code. Since then, the EGSnrc code has been released which has more accurate electron transport algorithms. We calculate the effect on the measured stopping powers of using EGSnrc instead of EGS4. METHOD: The EGS4 spectra calculated in PIRS-0626 were based on 4 × 10 5 primary electron histories. We first show that those spectra, calculated in 1997, are consistent with current EGS4 spectra calculated using 10 8 histories. EGSnrc spectra are also calculated using 10 8 histories and these high-precision spectra are compared to extract any energy difference. The energy differences between the spectra are used to estimate the effect on the measured electronic stopping powers. RESULTS: The energy differences depend on the absorber material, the absorber thickness and the beam energy. The improved electron elastic scattering cross section of EGSnrc accounts for only part of the difference between the two codes. The effect on the extracted stopping power is largest for the lowest electron energies and can be as large as 0.9%. The calculated spectra show differences for lower energies, with the EGSnrc spectra having a larger proportion of low-energy electrons. CONCLUSION: The differences introduced by using EGSnrc instead of EGS4 can affect the estimated stopping power by almost 1% in the worst case but generally the effect is much smaller. We report corrections that can be applied to all the stopping power data in PIRS-0626. An experiment to measure the average energy to create an ion pair in air, W air , using aluminum detectors will provide an interesting test of the aluminum stopping power data as reported in PIRS-0626 and revised by this work.

Determination of Wair in high-energy electron beams using graphite detectors.

PURPOSE: ICRU Report 90 on Key Data for Ionizing-Radiation Dosimetry: Measurement Standards and Applications (2014) has reaffirmed the recommended value of the mean energy required to create an ion pair in air, Wair , to be 33.97(12) eV. The report also indicates that this "constant" of radiation dosimetry is energy independent above 10 keV, since there is no theoretical or experimental evidence to the contrary. The goal of this investigation is to obtain additional experimental determinations of Wair in high energy beams and thus to verify the suggested energy independence. METHODS: Wair can be evaluated by combining ionometric and calorimetric measurements with a calculated ratio of the absorbed dose in the ion chamber air cavity and that of the calorimeter absorbing element. In this investigation, a graphite parallel plate chamber and a graphite calorimeter were used and the dose ratio was calculated using the EGSnrc Monte Carlo code. Measurements were made in electron beams from the NRC Vickers linear accelerator at two incident energies, 20 and 35 MeV. A range of average energies at the measurement point were obtained by inserting graphite plates in the primary beam. RESULTS: The average value of Wair obtained in this investigation is 33.85(18) eV which is consistent with the recommended value of 33.97(12) eV where the number in brackets represents the combined standard uncertainty of the value, referring to the corresponding last digits. The individual values of Wair do not show any statistically significant energy dependence. CONCLUSION: The overall combined uncertainty of 0.5% meets the original target of the investigation. A larger-scale investigation, involving more individual energy points and a wider range of electron energies is required to go further and, for example, comment on the Wair energy dependency question raised by Tessier et al. [Med. Phys. 2018;45:370-381].

Extracting Wair from the electron beam measurements of Domen and Lamperti.

PURPOSE: The average energy expended by an energetic electron to create an ion pair in dry air, Wair , is a key quantity in radiation dosimetry. Although Wair is well established for electron energies up to about 3 MeV, there is limited data for higher energies. The measurements by Domen and Lamperti [Med. Phys. 3, 294-301 (1976)] using electron beams in the energy range from 15 to 50 MeV can, in principle, be used to deduce values for Wair , if the electron stopping power of graphite and air are known. A previous analysis of these data revealed an anomalous variation of 2% in Wair as a function of the electron energy. We use Monte Carlo simulation techniques to reanalyze the original data and obtain new estimates for Wair , and to investigate the source of the reported anomaly. METHODS: Domen and Lamperti (DL) reported the ratio of the response of a graphite calorimeter to that of a graphite ionization chamber for broad beams of electrons with energies between 15 and 50 MeV and at different depths in graphite (including depths well beyond the range of the primary electrons, i.e., in the bremsstrahlung photon regime). Using a detailed EGSnrc model of the DL apparatus, as well as up-to-date stopping powers, we compute the dose ratio between the ionization chamber cavity and the calorimeter core, for plane-parallel electron beams. This dose ratio, multiplied by the DL measured ratio, provides a direct estimate for Wair . RESULTS: Despite an improved analysis of the original work, the extracted values of Wair still exhibit an increase as the mean electron energy at the point of measurement decreases below about 15 MeV. This anomalous trend is dubious physically, and inconsistent with extensive data for Wair obtained at lower energies. A thorough sensitivity analysis indicates that this trend is unlikely to stem from errors in extrapolation and correction procedures, uncertainties in electron stopping powers, or bias in calorimetry or ionization chamber measurements. However, we find that results are quite sensitive to the intrinsic graphite mass thickness of the detectors and to the incident beam energy. CONCLUSIONS: The DL experiment provides data in an energy regime where the electron stopping power is insensitive to the mean excitation energy of graphite - an issue plaguing Wair experiments at lower energies. Unfortunately, state-of-the-art scrutiny of the original data cannot explain the anomalous trend in terms of perturbation effects or extrapolation bias. It can only be understood in terms of speculative offsets in graphite mass thickness or beam energy. Therefore higher accuracy measurements for electron energies above 15 MeV are recommended to further resolve the value of Wair .

Electron beam water calorimetry measurements to obtain beam quality conversion factors.

PURPOSE: To provide results of water calorimetry and ion chamber measurements in high-energy electron beams carried out at the National Research Council Canada (NRC). There are three main aspects to this work: (a) investigation of the behavior of ionization chambers in electron beams of different energies with focus on long-term stability, (b) water calorimetry measurements to determine absorbed dose to water in high-energy beams for direct calibration of ion chambers, and (c) using measurements of chamber response relative to reference ion chambers, determination of beam quality conversion factors, kQ , for several ion chamber types. METHODS: Measurements are made in electron beams with energies between 8 MeV and 22 MeV from the NRC Elekta Precise clinical linear accelerator. Ion chamber measurements are made as a function of depth for cylindrical and plane-parallel ion chambers over a period of five years to investigate the stability of ion chamber response and for indirect calibration. Water calorimetry measurements are made in 18 MeV and 22 MeV beams. An insulated enclosure with fine temperature control is used to maintain a constant temperature (drifts less than 0.1 mK/min) of the calorimeter phantom at 4°C to minimize effects from convection. Two vessels of different designs are used with calibrated thermistor probes to measure radiation induced temperature rise. The vessels are filled with high-purity water and saturated with H2 or N2 gas to minimize the effect of radiochemical reactions on the measured temperature rise. A set of secondary standard ion chambers are calibrated directly against the calorimeter. Finally, several other ion chambers are calibrated in the NRC 60 Co reference field and then cross-calibrated against the secondary standard chambers in electron beams to realize kQ factors. RESULTS: The long-term stability of the cylindrical ion chambers in electron beams is better (always <0.15%) than plane-parallel chambers (0.2% to 0.4%). Calorimetry measurements made at 22 MeV with two different vessel geometries are consistent within 0.2% after correction for the vessel perturbation. Measurements of absorbed dose calibration coefficients for the same secondary standard chamber separated in time by 10 yr are within 0.2%. Drifts in linac output that would affect the transfer of the standard are mitigated to the 0.1% level by performing daily ion chamber normalization measurements. Calibration coefficients for secondary standard ion chambers can be achieved with uncertainties less than 0.4% (k = 1) in high-energy electron beams. The additional uncertainty in deriving calibration coefficients for well-behaved chambers indirectly against the secondary standard reference chambers is negligible. The kQ factors measured here differ by up to 1.3% compared to those in TG-51, an important change for reference dosimetry measurements. CONCLUSIONS: The measurements made here of kQ factors for eight plane-parallel and six cylindrical ion chambers will impact future updates of reference dosimetry protocols by providing some of the highest quality measurements of this crucial dosimetric parameter.

Direct measurement of electron beam quality conversion factors using water calorimetry.

PURPOSE: In this work, the authors describe an electron sealed water calorimeter (ESWcal) designed to directly measure absorbed dose to water in clinical electron beams and its use to derive electron beam quality conversion factors for two ionization chamber types. METHODS: A functioning calorimeter prototype was constructed in-house and used to obtain reproducible measurements in clinical accelerator-based 6, 9, 12, 16, and 20 MeV electron beams. Corrections for the radiation field perturbation due to the presence of the glass calorimeter vessel were calculated using Monte Carlo (MC) simulations. The conductive heat transfer due to dose gradients and nonwater materials was also accounted for using a commercial finite element method software package. RESULTS: The relative combined standard uncertainty on the ESWcal dose was estimated to be 0.50% for the 9-20 MeV beams and 1.00% for the 6 MeV beam, demonstrating that the development of a water calorimeter-based standard for electron beams over such a wide range of clinically relevant energies is feasible. The largest contributor to the uncertainty was the positioning (Type A, 0.10%-0.40%) and its influence on the perturbation correction (Type B, 0.10%-0.60%). As a preliminary validation, measurements performed with the ESWcal in a 6 MV photon beam were directly compared to results derived from the National Research Council of Canada (NRC) photon beam standard water calorimeter. These two independent devices were shown to agree well within the 0.43% combined relative uncertainty of the ESWcal for this beam type and quality. Absorbed dose electron beam quality conversion factors were measured using the ESWcal for the Exradin A12 and PTW Roos ionization chambers. The photon-electron conversion factor, kecal, for the A12 was also experimentally determined. Nonstatistically significant differences of up to 0.7% were found when compared to the calculation-based factors listed in the AAPM's TG-51 protocol. General agreement between the relative electron energy dependence of the PTW Roos data measured in this work and a recent MC-based study are also shown. CONCLUSIONS: This is the first time that water calorimetry has been successfully used to measure electron beam quality conversion factors for energies as low as 6 MeV (R50=2.25 cm).

The older adult experiencing sepsis.

Sepsis is a potentially fatal response to infection affecting patients across the life span. Sepsis can progress from systemic inflammatory response to severe sepsis and septic shock if not recognized promptly and managed effectively. Risk factors for sepsis include age, gender, the presence of invasive devices (eg, urinary catheters), and chronic medical conditions (eg, chronic obstructive pulmonary disease). Sepsis awareness is essential and includes identification of population-focused risk factors, recognition of clinical signs and symptoms, and timely implementation of interventions. The purpose of this article was to examine sepsis in older adults, including prevalence, atypical presentation of the condition, and considerations for sepsis management in the elderly population.

The Role of the Nurse Practitioner and Asymptomatic Urinary Treatments.

Asymptomatic urinary tract infections (aUTIs) are common among older adults in long-term care facilities (LTCFs) and studies have shown that they are inappropriately treated with antibiotics. We retrospectively characterized treatment strategies among 89 cases of aUTIs before and after a long-term facility hired a full-time nurse practitioner (NP). We found that residents with aUTIs were prescribed significantly more supportive treatment strategies after hiring an NP. However, there was no significant drop in the rate of inappropriate antibiotic treatments for aUTIs after hiring an NP.

The journey toward shared governance: the lived experience of nurse managers and staff nurses.

AIMS: The purpose of the study was to explore the lived experience of nurse managers and staff nurses in shared governance. BACKGROUND: Shared governance refers to systems and services aligned in partnership. The information gained by studying the lived experience of nurse managers and staff nurses in shared governance is valuable for providing knowledge of empowerment. METHODS: A qualitative design was used. Data were collected through a semi-structured interview using five questions with 11 Registered Nurses. Data were analysed through thematic analysis. RESULTS: Four themes emerged from data analysis. Nurse managers identified the journey of patient satisfaction; journey of empowerment; journey of self-management and journey of wellness. Staff nurses identified the journey of development and implementation of best practice; journey to provide quality patient care, journey to a new culture of nursing; and journey of a variety of challenges. CONCLUSIONS: This study supports the idea that collaboration between nurse managers and staff nurses develops a journey toward shared governance. IMPLICATIONS FOR NURSING MANAGEMENT: Nursing management can use findings to empower nurses to collaborate with nurse managers toward best practice. This adds to current knowledge that partnership of nurse managers and staff nurses, supports and encourages ownership in shared governance.

Millennial generation student nurses' perceptions of the impact of multiple technologies on learning.

AIM: To determine how millennial nursing students perceive the effects of instructional technology on their attentiveness, knowledge, critical thinking, and satisfaction. BACKGROUND Millennial learners develop critical thinking through experimentation, active participation, and multitasking with rapid shifts between technological devices. They desire immediate feedback. METHOD; A descriptive, longitudinal, anonymous survey design was used with a convenience sample of 108 sophomore, junior, and senior baccalaureate nursing students (participation rates 95 percent, winter, 85 percent, spring). Audience response, virtual learning, simulation, and computerized testing technologies were used. An investigator-designed instrument measured attentiveness, knowledge, critical thinking, and satisfaction (Cronbach's alphas 0.73, winter; 0.84, spring). RESULTS: Participants positively rated the audience response, virtual learning, and simulation instructional technologies on their class participation, learning, attention, and satisfaction. They strongly preferred computerized testing. CONCLUSION: Consistent with other studies, these students engaged positively with new teaching strategies using contemporary instructional technology. Faculty should consider using instructional technologies.

Human patient simulators and interactive case studies: a comparative analysis of learning outcomes and student perceptions.

Although human patient simulators provide an innovative teaching method for nursing students, they are quite expensive. To investigate the value of this expenditure, a quantitative, quasi-experimental, two-group pretest and posttest design was used to compare two educational interventions: human patient simulators and interactive case studies. The sample (N = 49) consisted of students from baccalaureate, accelerated baccalaureate, and diploma nursing programs. Custom-designed Health Education Systems, Inc examinations were used to measure knowledge before and after the implementation of the two educational interventions. Students in the human patient simulation group scored significantly higher than did those in the interactive case study group on the posttest Health Education Systems, Inc examination, and no significant difference was found in student scores among the three types of nursing programs that participated in the study. Data obtained from a questionnaire administered to participants indicated that students responded favorably to the use of human patient simulators as a teaching method.

Treatment head disassembly to improve the accuracy of large electron field simulation.

PURPOSE: The purposes of this study are to improve the accuracy of source and geometry parameters used in the simulation of large electron fields from a clinical linear accelerator and to evaluate improvement in the accuracy of the calculated dose distributions. METHODS: The monitor chamber and scattering foils of a clinical machine not in clinical service were removed for direct measurement of component geometry. Dose distributions were measured at various stages of reassembly, reducing the number of geometry variables in the simulation. The measured spot position and beam angle were found to vary with the beam energy. A magnetic field from the bending magnet was found between the exit window and the secondary collimators of sufficient strength to deflect electrons 1 cm off the beam axis at 100 cm from the exit window. The exit window was 0.05 cm thicker than manufacturer's specification, with over half of the increased thickness due to water pressure in the channel used to cool the window. Dose distributions were calculated with Monte Carlo simulation of the treatment head and water phantom using EGSnrc, a code benchmarked at radiotherapy energies for electron scatter and bremsstrahlung production, both critical to the simulation. The secondary scattering foil and monitor chamber offset from the collimator rotation axis were allowed to vary with the beam energy in the simulation to accommodate the deflection of the beam by the magnetic field, which was not simulated. RESULTS: The energy varied linearly with bending magnet current to within 1.4% from 6.7 to 19.6 MeV, the bending magnet beginning to saturate at the highest beam energy. The range in secondary foil offset used to account for the magnetic field was 0.09 cm crossplane and 0.15 cm inplane, the range in monitor chamber offset was 0.14 cm crossplane and 0.07 cm inplane. A 1.5%/0.09 cm match or better was obtained to measured depth dose curves. Profiles measured at the depth of maximum dose matched the simulated profiles to 2.6% or better at doses of 80% or more of the dose on the central axis. The profiles along the direction of MLC motion agreed to within 0.16 cm at the edge of the field. There remained a mismatch for the lower beam energies at the edge of the profile that ran parallel to the direction of jaw motion of up to 1.4 cm for the 6 MeV beam, attributed to the MLC support block at the periphery of the field left out of the simulation and to beam deflection by the magnetic field. The possibility of using these results to perform accurate simulation without disassembly is discussed. Phase-space files were made available for benchmarking beam models and other purposes. CONCLUSIONS: The match to measured large field dose distributions from clinical electron beams with Monte Carlo simulation was improved with more accurate source details and geometry details closer to manufacturer's specification than previously achieved.

Benchmarking of Monte Carlo simulation of bremsstrahlung from thick targets at radiotherapy energies.

Several Monte Carlo systems were benchmarked against published measurements of bremsstrahlung yield from thick targets for 10-30 MV beams. The quantity measured was photon fluence at 1 m per unit energy per incident electron (spectra), and total photon fluence, integrated over energy, per incident electron (photon yield). Results were reported at 10-30 MV on the beam axis for Al and Pb targets and at 15 MV at angles out to 90 degrees for Be, Al, and Pb targets. Beam energy was revised with improved accuracy of 0.5% using an improved energy calibration of the accelerator. Recently released versions of the Monte Carlo systems EGSNRC, GEANT4, and PENELOPE were benchmarked against the published measurements using the revised beam energies. Monte Carlo simulation was capable of calculation of photon yield in the experimental geometry to 5% out to 30 degrees, 10% at wider angles, and photon spectra to 10% at intermediate photon energies, 15% at lower energies. Accuracy of measured photon yield from 0 to 30 degrees was 5%, 1 s.d., increasing to 7% for the larger angles. EGSNRC and PENELOPE results were within 2 s.d. of the measured photon yield at all beam energies and angles, GEANT4 within 3 s.d. Photon yield at nonzero angles for angles covering conventional field sizes used in radiotherapy (out to 10 degrees), measured with an accuracy of 3%, was calculated within 1 s.d. of measurement for EGSNRC, 2 s.d. for PENELOPE and GEANT4. Calculated spectra closely matched measurement at photon energies over 5 MeV. Photon spectra near 5 MeV were underestimated by as much as 10% by all three codes. The photon spectra below 2-3 MeV for the Be and Al targets and small angles were overestimated by up to 15% when using EGSNRC and PENELOPE, 20% with GEANT4. EGSNRC results with the NIST option for the bremsstrahlung cross section were preferred over the alternative cross section available in EGSNRC and over EGS4. GEANT4 results calculated with the "low energy" physics list were more accurate than those calculated with the "standard" physics list.

Oil and gas projects in the Western Amazon: threats to wilderness, biodiversity, and indigenous peoples.

BACKGROUND: The western Amazon is the most biologically rich part of the Amazon basin and is home to a great diversity of indigenous ethnic groups, including some of the world's last uncontacted peoples living in voluntary isolation. Unlike the eastern Brazilian Amazon, it is still a largely intact ecosystem. Underlying this landscape are large reserves of oil and gas, many yet untapped. The growing global demand is leading to unprecedented exploration and development in the region. METHODOLOGY/PRINCIPAL FINDINGS: We synthesized information from government sources to quantify the status of oil development in the western Amazon. National governments delimit specific geographic areas or "blocks" that are zoned for hydrocarbon activities, which they may lease to state and multinational energy companies for exploration and production. About 180 oil and gas blocks now cover approximately 688,000 km(2) of the western Amazon. These blocks overlap the most species-rich part of the Amazon. We also found that many of the blocks overlap indigenous territories, both titled lands and areas utilized by peoples in voluntary isolation. In Ecuador and Peru, oil and gas blocks now cover more than two-thirds of the Amazon. In Bolivia and western Brazil, major exploration activities are set to increase rapidly. CONCLUSIONS/SIGNIFICANCE: Without improved policies, the increasing scope and magnitude of planned extraction means that environmental and social impacts are likely to intensify. We review the most pressing oil- and gas-related conservation policy issues confronting the region. These include the need for regional Strategic Environmental Impact Assessments and the adoption of roadless extraction techniques. We also consider the conflicts where the blocks overlap indigenous peoples' territories.

Nicaragua: an example of commitments and strengths despite problems of poverty.

Nicaragua is located in the middle of the Central American isthmus between the countries of Honduras and Costa Rica. It is the largest Central American country and is equivalent in size to the state of Georgia. Nicaragua is cited by Pan American Health Organization as one of the poorest third-world countries. One factor that continues to contribute to Nicaragua's chronic poverty state is the demographics of the country. Nearly half of all Nicaraguans are under 15 years of age, and more than a quarter are between the ages of 15 and 29 years. Only a quarter of the population is over 30 years of age. Beyond the hardship and poverty, there is a country rich in beauty. Nicaragua has a beautiful countryside with lush green mountains, black sand beaches of the Pacific Ocean, and the natural wonder of active volcanoes. It is easy to become engulfed by the tranquility of these surroundings and to steer away from the harsh conditions of the country. It is, however, a temporary escape from reality, for it was the hardships and unfavorable circumstances of this country that are never forgotten and which persist until today. This article focuses on a variety of interventions used to assist Nicaragua with their health care and state of well-being.

Experimental determination of beam quality factors, kQ, for two types of Farmer chamber in a 10 MV photon and a 175 MeV proton beam.

Absorbed doses determined with a sealed water calorimeter operated at 4 degrees C are compared with the results obtained using ionization chambers and the IAEA TRS-398 code of practice in a 10 MV photon beam (TPR(20,10) = 0.734) and a 175 MeV proton beam (at a depth corresponding to the residual range, R(res) = 14.7 cm). Three NE 2571 and two FC65-G ionization chambers were calibrated in terms of absorbed-dose-to-water in (60)Co at the Swedish secondary standard dosimetry laboratory, directly traceable to the BIPM. In the photon beam quality, calorimetry was found to agree with ionometry within 0.3%, confirming the k(Q) values tabulated in TRS-398. In contrast, a 1.8% deviation was found in the proton beam at 6 g cm(-2) depth, suggesting that the TRS-398 tabulated k(Q) values for these two ionization chamber types are too high. Assuming no perturbation effect in the proton beam for the ionization chambers, a value for (w(air)/e)(Q) of 33.6 J C(-1) +/- 1.7% (k = 1) can be derived from these measurements. An analytical evaluation of the effect from non-elastic nuclear interactions in the ionization chamber wall indicates a perturbation effect of 0.6%. Including this estimated result in the proton beam would increase the determined (w(air)/e)(Q) value by the same amount.

An investigation of the photon energy dependence of the EPR alanine dosimetry system.

The electron paramagnetic resonance (EPR) alanine dosimetry system is based on EPR measurements of radicals formed in alanine by ionizing radiation. The system has been studied to determine its energy dependence for photons in the 10-30 MV region relative to those of 60Co and to find out if the system would be suitable for dosimetry comparisons. The irradiations were carried out at the National Research Council, Ottawa, Canada and the doses ranged from 8 to 54 Gy. The EPR measurements were performed at the University of Oslo, Norway. The ratio of the slope of the alanine reading versus dose-to-water curve for a certain linac photon beam quality and the corresponding slope for a reference 60Co gamma-radiation gives an experimental measure of the relative dose-to-water response of the EPR alanine dosimetry system. For calculating the linear regression coefficients of these alanine reading versus dose curves, the method of weighted least squares was used. This method is assumed to produce more accurate regression coefficients when applied to EPR dosimetry than the common method of standard least squares. The overall uncertainty on the ratio of slopes was between 0.5 and 0.6% for all three linac energies. The relative response for all the linac beams compared to cobalt was less than unity: by about 0.5% for the 20 and 30 MV points but by more than 1% for the 10 MV point. The given standard uncertainties negate concluding that there is any significant internal variation in the measured response as a function of beam quality between the three linac energies. Thus, we calculated the average dose response for all three energies and found that the alanine response is 0.8% (+/-0.5%) lower for high energy x-rays than for 60Co gamma-rays. This result indicates a small energy dependence in the alanine response for the high-energy photons relative to 60Co which may be significant. This result is specific to our dosimetry system (alanine with 20% polyethylene binder pressed into a particular shape) including its waterproofing sleeve of PMMA (2 mm thick); however, we expect that this result may apply to other similar detectors.

Evaluation of an ion-exchange membrane for the purification of plasmid DNA.

Separation and purification of large quantities of plasmid DNA (pDNA) is a particularly difficult manufacturing issue because of the relatively low capacity, flow rate and purity observed using traditional bead-based chromatography. The objective of the present study was to evaluate the performance of anion-exchange membranes for the purification of pDNA from Escherichia coli lysate solution. The fate of host-cell protein and endotoxin relative to pDNA was measured and used to calculate recoveries, mass balances, dynamic capacities and purification factors as a function of the flow rate and loading volume of the lysate solution. Breakthrough curves were not sigmoidal and symmetric in shape. They rose sharply at first, and then slowly towards, but never reaching, saturation. Conversely, elution curves were independent of flow rate. pDNA bound tightly to the membranes, whereas protein and endotoxin did not. Dynamic binding capacity for pDNA was 20-25 times greater, and the flow rate was 55-550 times greater, than values observed for beads. However, some pDNA bound irreversibly to the membrane surface and was not removed completely during elution. The intrinsic rate of pDNA adsorption to the membrane was found to be rate-limiting, whereas effects of liquid-phase mass transfer and flow non-idealities were negligible. These results were interpreted using models of adsorption that included steric effects using the 'car-parking-problem' model, and surface residence time effects using the spreading model. This work demonstrated the advantages of ion-exchange membranes compared with beads for the purification of large biomolecules such as pDNA.

Water Calorimetry: A Correction to the Heat Defect Calculations.

In a recent publication, we used a reaction model (model III) to calculate the heat defect for the irradiation of aqueous solutions with ionizing radiation at 21 °C. Subsequent work has revealed that the literature value used for one of the rate constants in the model was incorrect. A revised model (model IIIR) incorporates the correct rate constant for 21 °C. Versions of models III and IIIR were created for irradiations at 4 °C. For our current water calorimetry protocol, the values of the heat defect for H2/O2-water (water saturated with a flow of 43 % H2 and 57 % O2, by volume) at 21 °C predicted by model III and model IIIR are similar but the value for 4 °C predicted by III is 30 % smaller than the value predicted by IIIR. Model IIIR predicts that the values of the heat defect at 21 °C and 4 °C lie within the range -0.023±0.002, in agreement with the values obtained from our water calorimetry measurements done using pure water and H2-saturated water at 21 °C and 4 °C. The yields of hydrogen peroxide in H2/O2-water at 21 °C and 4 °C were measured and agree with the predictions of model IIIR. Our water calorimetry measurements made with pure water and H2-saturated water are now of sufficient quality that they can be used to determine the heat defect for H2/O2-water better than can be done by simulations. However, consistency between the three systems continues to be an excellent check on water purity which is crucial, especially for the pure water system.