Non-chemotherapy adjuvant agents in TP53 mutant Ewing sarcoma.

Ewing sarcoma (EWS) is a malignant tumor arising in bone or soft tissue that occurs in adolescent and young adult patients as well as adults later in life. Although non-metastatic EWS is typically responsive to treatment when newly diagnosed, relapsed cases have an unmet need for which no standard treatment approach exists. Recent phase III clinical trials for EWS comparing 7 vs 5 chemotherapy drugs have failed to improve survival. To extend the durability of remission for EWS, we investigated 3 non-chemotherapy adjuvant therapy drug candidates to be combined with chemotherapy. The efficacy of these adjuvant drugs was investigated via anchorage-dependent growth assays, anchorage-independent soft-agar colony formation assays and EWS xenograft mouse models. Enoxacin and entinostat were the most effective adjuvant drug in both long-term in vitro and in vivo adjuvant studies. In the context that enoxacin is an FDA-approved antibiotic, and that entinostat is an investigational agent not yet FDA-approved, we propose enoxacin as an adjuvant drug for further preclinical and clinical investigation in EWS patients.

High sustained virologic response rates, regardless of race or socioeconomic class, in patients treated with chronic hepatitis C in community practice using a specialized pharmacy team.

Approved direct-acting antiviral (DAA) regimens against hepatitis C virus (HCV) can cure nearly all patients; however, socioeconomic disparities may impact access and outcome. This study assesses socioeconomic factors, differences in insurance coverage and the drug prior authorization process in HCV-infected patients managed in community practices partnered with a dedicated pharmacy team with expertise in liver disease. This Institutional Review Board-approved, ongoing study captures data on a cohort of 2480 patients from community practices. Patients had chronic hepatitis C and were treated with DAA regimens selected by their physician. The HCV Health Outcomes Centers Network provides comprehensive patient management including a dedicated pharmacy support team with expertise in the prior authorization process. In this cohort, 60.1% were male, 49% were Hispanic Whites (HW), 37% were Non-Hispanic Whites (NHW), and 14% were Black/African American (BAA). Eighty-seven percent of patients were treatment-naïve, 74% were infected with genotype 1 virus and 63% had advanced fibrosis/cirrhosis (F3/F4 = 68.2% HW, 65.6% BAA, 55.4% NHW). Forty percent of patients were on disability with the highest percentage in the BAA group and less than one-third were employed full time, regardless of race/ethnicity. Medicare covered 42% of BAA patients versus 32% of HW and NHW. The vast majority of HW (80%) and BAA (75%) had a median income below the median income of Texas residents. Additionally, 75% of HW and 71% of BAA had median income below the poverty level in Texas. Despite the above socioeconomic factors, 92% of all prior authorizations were approved upon first submission and patients received DAAs an average of 17 days from prescription. DAA therapy resulted in cure in 95.3% of patients (sustained virologic response = 94.8% HW, 94.0% BAA, 96.5% NHW). Despite having more advanced diseases and more negative socioeconomic factors, >94% of HW and BAA patients were cured. Continued patient education and communication with the healthcare team can lead to high adherence and > 94% HCV cure rates regardless of race/ethnicity or underlying socioeconomic factors in the community setting.

Uptake of internet-delivered UK adult hearing assessment.

OBJECTIVE: To evaluate uptake of the internet-based hearing test, with respect to the 11% of UK adults that have hearing loss but do not use hearing aids. DESIGN: Feasibility study in a primary care practice in the North of England. STUDY SAMPLE: Adults aged 50-74 years were sent postal invitations to complete an internet hearing test (N = 600). Those who completed the test, those who failed (>35 dB HL in the better ear) and demographic correlates (age, gender, ethnicity and socioeconomic level) were recorded. RESULTS: 11.2% of invited adults completed the hearing test and 7.7% failed it. Those who took the test tended to have a higher socioeconomic background than those who did not. There were no differences in age, ethnicity or gender between those who took the test and those who did not. CONCLUSIONS: An estimated 70% (7.7%/11.0%) of adults with hearing loss but who do not use hearing aids took the test. Uptake was equitable across most demographic categories. Uptake was high among a study sample that was substantially more deprived than the general UK population. Internet-based hearing testing offers an efficient paradigm for identifying hearing loss.

Real-world observational experience with direct-acting antivirals for hepatitis C: baseline resistance, efficacy, and need for long-term surveillance.

The aim of this study was to obtain real-world, US, observational data on the effect of baseline resistance-associated substitutions (RASs) on achieving sustained virologic response (SVR) in hepatitis C (HCV) patients treated with direct-acting antiviral (DAA) regimens; the need for long-term follow-up in post-SVR patients.It is uncertain if the presence of RASs limits efficacy to DAAs. Once SVR is achieved, society guidelines recommend long-term surveillance for hepatocellular carcinoma in certain patients. Real-world data are limited on these topics.Adult patients treated with DAAs at community hepatitis clinics between January 2015 and April 2017 were included in this study. Baseline resistance testing was performed before treatment. Per guidelines, post-SVR long-term monitoring was required in patients with F3 to F4 fibrosis before treatment or with elevated ALT levels (>19 U/L females; >30 U/L males).A total of 875 chronic, mostly GT1a (60%) HCV patients were treated with an approved DAA regimen. Average baseline AST and ALT were 75 and 67 U/L, respectively, and 47% had F3 to F4 fibrosis at baseline. SVR was achieved in 863 (98.6%) patients despite a high presence of baseline RASs (61%). Long-term monitoring was required post-SVR in 539 patients (62%).In a real-life, US cohort of HCV-infected patients, nearly all patients achieved SVR with available DAA regimens regardless of baseline RASs. Approximately two-thirds of these patients required long-term follow-up, despite viral eradication.

Transesterification of propylene glycol methyl ether in chromatographic reactors using anion exchange resin as a catalyst.

Reactive chromatography using an anion exchange resin is proposed for a transesterification reaction of propylene glycol methyl ether (DOWANOL™ PM) with ethyl acetate to produce propylene glycol methyl ether acetate (DOWANOL™ PMA). This reaction is studied in batch and chromatographic reactors catalyzed by an anion exchange resin. Several anion exchange resins are tested and compared based on the performance of resin as an adsorbent and a catalyst. A chromatographic column is packed with a selected catalyst, AMBERLITE™ IRA904, and both reaction and chromatographic elution are studied at different temperatures and feed concentrations. The resulting chromatograms are fitted to a mathematical model to obtain adsorption equilibrium and reaction kinetic parameters by the inverse method. Compared to esterification investigated in a previous study, transesterification has advantages such as a higher conversion at lower temperature and easy removal of the byproduct which may lead to higher productivity. Deactivation of anion exchange resins is observed and potential solutions are suggested.

Optimization of reactive simulated moving bed systems with modulation of feed concentration for production of glycol ether ester.

In this article, we extend the simulated moving bed reactor (SMBR) mode of operation to the production of propylene glycol methyl ether acetate (DOWANOL™ PMA glycol ether) through the esterification of 1-methoxy-2-propanol (DOWANOL™ PM glycol ether) and acetic acid using AMBERLYST™ 15 as a catalyst and adsorbent. In addition, for the first time, we integrate the concept of modulation of the feed concentration (ModiCon) to SMBR operation. The performance of the conventional (constant feed) and ModiCon operation modes of SMBR are analyzed and compared. The SMBR processes are designed using a model based on a multi-objective optimization approach, where a transport dispersive model with a linear driving force for the adsorption rate has been used for modeling the SMBR system. The adsorption equilibrium and kinetics parameters are estimated from the batch and single column injection experiments by the inverse method. The multiple objectives are to maximize the production rate of DOWANOL™ PMA glycol ether, maximize the conversion of the esterification reaction and minimize the consumption of DOWANOL™ PM glycol ether which also acts as the desorbent in the chromatographic separation. It is shown that ModiCon achieves a higher productivity by 12-36% over the conventional operation with higher product purity and recovery.

Does BCG vaccination protect against childhood asthma? Final results from the Manchester Community Asthma Study retrospective cohort study and updated systematic review and meta-analysis.

BACKGROUND: The Manchester Community Asthma Study (MANCAS) found a protective effect against the risk of wheeze at age 6 to 11 years for children given neonatal BCG vaccination. Our subsequent systematic review and meta-analysis suggested that BCG vaccination did not protect against allergic sensitization but might have exerted a protective effect against nonatopic asthma. OBJECTIVES: We sought to assess whether the protective effect of BCG vaccination on wheeze observed in the MANCAS cohort was maintained at age 13 to 17 years and to incorporate the findings from this final MANCAS analysis into an updated systematic review and meta-analysis. METHODS: BCG vaccination status was determined from health records and respiratory outcomes from questionnaire responses. We updated the systematic review and used fixed-effects and random-effects modeling to undertake meta-analyses. RESULTS: There were 1608 participants in the final MANCAS analysis. The 12-month prevalence of wheeze was 15.1%. There was no difference in prevalence between those who were and were not BCG vaccinated (15.8% vs 14.3%; relative risk, 1.05; 95% CI, 0.94-1.19). The updated meta-analysis incorporated 4 new studies: this showed that the protective effect of BCG vaccination against the development of asthma identified in our previous meta-analysis was attenuated (odds ratio, 0.95; 95% CI, 0.89-1.00). No protective effect of BCG was seen for sensitization, eczema/atopic dermatitis, rhinoconjunctivitis, or allergy in general. CONCLUSIONS: Taken together, the final results of the MANCAS cohort and the updated systematic review and meta-analysis provide clearer evidence that any protective effect of BCG vaccination on childhood asthma is likely to be transient.

Making audit actionable: an example algorithm for blood pressure management in chronic kidney disease.

Despite widespread use of clinical guidelines, actual care often falls short of ideal standards. Electronic health records (EHR) can be analyzed to provide information on how to improve care, but this is seldom done in sufficient detail to guide specific action. We developed an algorithm to provide practical, actionable information for care quality improvement using blood pressure (BP) management in chronic kidney disease (CKD) as an exemplar. We used UK clinical guidelines and EHR data from 440 patients in Salford (UK) to develop the algorithm. We then applied it to 532,409 individual patient records, identifying 11,097 CKD patients, 3,766 (34%) of which showed room for improvement in their care: either through medication optimization or better BP monitoring. Manual record reviews to evaluate accuracy indicated a positive-predictive value of 90%. Such algorithms could help improve the management of chronic conditions by providing the missing link between clinical audit and decision support.

Prevalence of respiratory symptoms at two time points in a population of children in Manchester--a cohort study.

OBJECTIVE: Although the prevalence of asthma and atopy has been noted to have increased in recent decades, patterns of asthma prevalence have, traditionally, been difficult to track. Most reports on trends in childhood asthma have been cross-sectional measuring the prevalence in cohorts of similar aged children at different time points. The aim of this paper is to report on the prevalence of symptoms in the same cohort at two separate time points. DESIGN: Retrospective cohort study. SETTING: Community-based study, Central Manchester. PARTICIPANTS: MANCAS1, study n=5086, participation n=2414. MANCAS2, study n=6338, participation n=1608. Children born in a hospital in Manchester within specified dates and still living or attending a school in Central Manchester were eligible for inclusion. Children on an 'at-risk' register or living with short-term carers were excluded. OUTCOME MEASURES: Data on respiratory symptoms were collected at two separate time points using parent completed questionnaires. RESULTS: Response rate for MANCAS1 was 47.5% and 25.4% for MANCAS2. There were 801 individuals for whom a response to both studies was received. There was a significant reduction in the prevalence of night cough (29.5% vs 18.3%, McNemar <0.01) and antibiotic use for respiratory infections (9.1% vs 4.3%, McNemar <0.01) between the two study time points. The prevalence of hay fever/eczema increased (41.6% vs 46.9%, McNemar <0.01) between the two studies. There was no significant difference in the prevalence of wheeze, exercise-induced wheeze or asthma medication. CONCLUSIONS: Although this report of respiratory symptom prevalence in the same population at two time points over a 7-year period shows a constant burden of asthma symptoms, there is some suggestion of variability in asthma symptom prevalence within the cohort as the children matured while the burden of allergy symptoms increased.

Standards of care for occupational asthma: an update.

BACKGROUND: The British Thoracic Society (BTS) Standards of Care (SoC) Committee produced a standard of care for occupational asthma (OA) in 2008, based on a systematic evidence review performed in 2004 by the British Occupational Health Research Foundation (BOHRF). METHODS: BOHRF updated the evidence base from 2004-2009 in 2010. RESULTS: This article summarises the changes in evidence and is aimed at physicians, nurses and other healthcare professionals in primary and secondary care, occupational health and public health and at employers, workers and their health, safety and other representatives. CONCLUSIONS: Various recommendations and evidence ratings have changed in the management of asthma that may have an occupational cause.

Temporal change in health-related quality of life: a longitudinal study in general practice 1999-2004.

BACKGROUND: In order to assess and plan for changing healthcare needs, the lack of available information regarding temporal changes in the health-related quality of life of a population must be addressed. AIM: This paper aims to describe such changes over 5 years in a general population. DESIGN OF STUDY: Longitudinal postal questionnaire study. SETTING: UK general practice. METHOD: This was a longitudinal postal questionnaire study in two general practice populations, using the generic instrument EQ-5D to measure health-related quality of life. Individuals were included if they responded to three postal surveys in 1999, 2001, and 2004 and there were three consecutive values of EQ-5D(index) available between 1999 and 2004. RESULTS: A total of 2498 subjects were included in the study. After adjustment for potential confounders (including ageing), health-related quality of life declined significantly over the observation period. The change in EQ-5D(index) was from 0.79 to 0.74 and for EQ-5D(vas) 76.8 to 73.3 (P for both trends <0.001). CONCLUSION: Health-related quality of life deteriorated in these populations over 5 years. In an era of improvements in mortality, this has important implications for the use of health-related quality of life data in healthcare planning and resource allocation.

How do common chronic conditions affect health-related quality of life?

BACKGROUND: Comorbidity is common. National Institute for Health and Clinical Excellence (NICE) guidelines have been created to make best use of resources to improve patients' quality of life but do not currently take account of comorbidity. The effect of multiple chronic conditions with regard to health-related quality of life (HRQoL) is poorly researched. Criticisms of previous research have been due to patient-defined chronic conditions, lack of quantification of the effects of confounding factors, selection of affected patients only, small sample sizes, and upper age limits. AIM: This study aims to address these issues, looking into the impact of combinations of chronic conditions on HRQoL. DESIGN OF THE STUDY: Participants filled in a questionnaire containing general health information, specific respiratory questions, and the EQ-5D measure of HRQoL. The questionnaires were then matched up to their GP records to obtain their disease status for six common chronic diseases (asthma, chronic obstructive pulmonary disease, ischaemic heart disease, hypertension, diabetes, and cerebrovascular disease). METHOD: Data from a mailed questionnaire were analysed from 5169 patients aged >16 years from two general practices in Wythenshawe, Manchester in 2004. Completion of the questionnaire was taken to indicate consent to participate. RESULTS: Significant correlations were found between a lower HRQoL and increasing numbers of chronic conditions (P<0.001), increasing age, possible obstructive airway disease, lack of higher education, smoking, and female sex. These all remained significant following regression, except for sex, with number of chronic conditions being a strong predictor of the weighted health state index score, EQ-5D(index) (coefficient = -0.079, P<0.001). CONCLUSION: Increasing numbers of chronic conditions have a strong negative effect on HRQoL.

Prevalence of airflow obstruction in patients attending a rapid access chest pain clinic.

BACKGROUND: Many UK hospitals have set-up specialised chest pain clinics to deal promptly and efficiently with cases of possible cardiac chest pain. It is possible that a proportion of patients attending these clinics will have a respiratory cause for their chest pain, or respiratory disease in addition to their cardiac pain. This study aimed to determine the prevalence of airflow obstruction, ischaemic heart disease and dual pathology in such patients. METHODS: Spirometry was performed on patients referred to a rapid access chest pain clinic over a 12-month period (target population of 400 patients). The main outcome measure was the prevalence of airflow obstruction (defined using spirometry), ischaemic heart disease and dual pathology. RESULTS: 405 subjects participated in the study. Abnormal spirometry was detected in 21% of patients (n=85). Airflow obstruction was the predominant lung function abnormality and was detected in 60 patients. Ischaemic heart disease was diagnosed in 21% of patients (n=85). Dual pathology was found in 4% of patients (n=17). CONCLUSIONS: Previous studies have reported a link between impaired lung function and future cardiovascular morbidity and mortality. This study suggests that airflow obstruction is an important alternative differential diagnosis in patients referred to a rapid access chest pain clinic. The identification of abnormal spirometry may help to better risk-stratify patients for future cardiovascular events and allow interventions to be instituted.

Prevalence of respiratory symptoms, features of asthma, and characteristics associated with respiratory disease, in 6-11 year olds in Manchester.

AIMS: This paper describes the prevalence of respiratory symptoms, features of asthma, and characteristics associated with respiratory disease in 6-11 year old children in an historical cohort study. METHODS: The study included 5086 children, all born in the same maternity unit in the north west of England over a four-year period. The prevalence of respiratory symptoms, features of asthma, and characteristics associated with respiratory disease were determined by the use of parent-completed questionnaires. Skin prick tests were used to ascertain atopic status. RESULTS: The response was 47.5%. The prevalence of wheeze, asthma medication use and atopic sensitisation were 20.3%, 16.2% and 37.1% respectively. Wheeze and atopy were significantly more prevalent in boys (22.4% versus 17.9% and 43.0% versus 29.3%, respectively). CONCLUSIONS: This study identified a high prevalence of respiratory disease in this population and provides a baseline for monitoring trends in respiratory disease in 6-11 year old children.

Factors influencing the response to postal questionnaire surveys about respiratory symptoms.

BACKGROUND: Response rates to postal questionnaires have been falling in recent years. AIM: To examine factors affecting the response to five postal respiratory questionnaire surveys. DESIGN: Cross sectional study. SETTING: General practice. METHOD: Five surveys were conducted in all adults registered with two UK general practices using an ECRHQ-based questionnaire, with two reminders at 4-week intervals. RESULTS: Response rates declined over time (1993 - 71.2%; 1995 - 70.5%; 1999 - 65.5%; 2001 - 65.3%; 2004 - 46.9%). Age and gender of non-responders were available for 2001 and 2004: responders were older (mean 48.8 years vs 37.6, p<0.001; 50.5 vs 38.8, p<0.001) and more likely to be female (54.9% vs 44.9%, p<0.001; 55.3% vs 48.5%, p<0.001). The response rate was increased by 18% (2004) and 23% (2001) by the use of two reminders. Early responders were older and more likely to be females, but were less likely to smoke than late responders after reminders. There was no important association between respiratory symptoms and associated feature prevalence and stage of response. CONCLUSION: Declining response rates may represent reduced motivation and reluctance to share personal information. Qualitative exploration of late/non-response could help reduce bias when planning and analysing such surveys. The use of two reminders is an important factor in improving response.

Long term prognosis in preschool children with wheeze: longitudinal postal questionnaire study 1993-2004.

OBJECTIVES: To follow a population of preschool children with and without parent reported wheeze over a period of 6-11 years to determine prognosis and its important predictive factors. DESIGN: Longitudinal series of five postal surveys based on the international study of asthma and allergies in childhood questionnaire carried out between 1993 and 2004. SETTING: Two general practice populations, south Manchester. PARTICIPANTS: 628 children aged less than 5 years at recruitment and those with at least six years' follow-up data. MAIN OUTCOME MEASURES: Parent completed questionnaire data for respiratory symptoms and associated features. RESULTS: Of 628 children included in the study, 201 (32%) had parent reported wheeze at the first observation (baseline), of whom 27% also reported the symptom on the second occasion (persistent asthma). The only important baseline predictors of persistent asthma were exercise induced wheeze (odds ratio 3.94, 95% confidence interval 1.72 to 9.00) and a history of atopic disorders (4.44, 1.94 to 10.13). The presence of both predictors indicated a likelihood of 53.2% of developing asthma; if only one feature was present this decreased to 17.2%, whereas if neither was present the likelihood was 10.9%. Family history of asthma was not predictive of persistent asthma among children with preschool wheeze. CONCLUSION: Using two simple predictive factors (baseline parent reported exercise induced wheeze and a history of atopic disorders), it is possible to estimate the likelihood of future asthma in children presenting with preschool wheeze. The absence of baseline exercise induced wheeze and a history of atopic disorders reduces the likelihood of subsequent asthma by a factor of five.

Dynamic response to heat--a novel physical characteristic of breast cancer.

We report a novel observation about breast cancer. We imaged 6 fresh pathological specimens of breast cancer (1 mastectomy and 5 lumpectomies) with a sensitive infrared camera. We found that, following exposure to hot air for 1min, the cancers warmed up much less than the surrounding normal breast with a difference ranging from 12 to 20 degrees C. We believe that this physical property of cancer should be explored and may give us better insight into the nature of cancer, its diagnosis (including intraoperative diagnosis) and targeted treatment.

Retraction by surface ferromagnetisation of target tissues: preliminary studies on feasibility of magnetic retraction for endoscopic surgery.

BACKGROUND: Magnetic retraction has potential advantages over existing direct physical retraction means (e.g., forceps) in terms of providing complete atraumatic retraction, avoiding tumour cell exfoliation as well as offering the possibility of noncontact retraction. This paper describes a pilot study of surface magnetic retraction of the gastric mucosa to facilitate resection. METHODS: Fifteen porcine stomach specimens were used in this pilot study. The uniaxial tensile properties of retracted mucosa were initially studied using a tensiometer. Magnetic media of ferromagnetic microparticles (stainless steel 410) dispersed in cyanoacrylate liquid were prepared at four different concentrations, and a neodymium permanent magnet was used to magnetically retract the media. The media was finally surface-glued to the target mucosa for performing a simulated surgical procedure. RESULTS: The force measurement data show that the retraction forces increased as the concentrations of microparticles and magnetic media volumes increased. A magnetic media concentration of 2 g/mL was most suitable since it offered sufficient retraction force from a small volume of applied media, e.g., the observed magnetic forces exerted on 50 microL of media were 1.42 N by a 3-mm magnet and 3.75 N by a 6-mm magnet, respectively, both being more than sufficient for the mucosal retraction. The additional forces required for dissection with four alternative instruments, i.e., electrosurgery hook, snares, scalpel or scissors, were also measured, e.g., the total force required to retract up to 10 mm and resect the mucosa with snares was 0.36 +/- 0.17 N. In a simulated surgical procedure (resection of gastric mucosa with glued magnetic medium) retraction by the magnet allowed resection of the tented mucosa by an electrosurgical snare. CONCLUSION: Surface ferromagnetisation of target mucosal tissues could enable magnetic retraction for endoscopic surgery.

National survey on the roles and training of primary care respiratory nurses in the UK in 2006: are we making progress?

OBJECTIVES: To describe nurse-led UK general practice asthma and chronic obstructive pulmonary disease (COPD) care, and the training undertaken to support it. METHODS: Questionnaires were sent to 500 randomly-selected UK asthma and COPD practice nurses. RESULTS: 382 nurses (76%) completed the practice characteristics section, 389 (78%) described their asthma roles and training, and 368 (74%) described their COPD roles and training. 96 practices (25%; 95%CI 21-29%) ran designated asthma clinics, 87 (23%; 95%CI 19- 27%) ran designated COPD clinics, and 170 (45%; 95%CI 40-49%) did not run designated respiratory clinics. Of the 255 nurses with an advanced asthma role, 51 (20%; 95%CI 15-25%) did not have accredited asthma training. Of the 215 nurses with an advanced COPD role, 111 (52%; 95%CI 45-58%) did not have accredited COPD training. CONCLUSION: Patients are increasingly being seen outside of designated asthma or COPD clinics, often by nurses with an advanced role. It is important that nurses have the training to fulfil this role.

Use of glycol ethers for selective release of periplasmic proteins from Gram-negative bacteria.

Genetic modification of Gram-negative bacteria to express a desired protein within the cell's periplasmic space, located between the inner cytoplasmic membrane and the outer cell wall, can offer an attractive strategy for commercial production of therapeutic proteins and industrial enzymes. In certain applications, the product expression rate is high, and the ability to isolate the product from the cell mass is greatly enhanced because of the product's compartmentalized location within the cell. Protein release methods that increase the permeability of the outer cell wall for primary recovery, but avoid rupturing the inner cell membrane, reduce contamination of the recovered product with other host cell components and simplify final purification. This article reports representative data for a new release method employing glycol ether solvents. The example involves the use of 2-butoxyethanol (commonly called ethylene glycol n-butyl ether or EB) for selective release of a proprietary biopharmaceutical protein produced in the periplasmic space of Pseudomonas fluorescens. In this example, glycol ether treatment yielded approximately 65% primary recovery with approximately 80% purity on a protein-only basis. Compared with other methods including heat treatment, osmotic shock, and the use of surfactants, the glycol ether treatment yielded significantly reduced concentrations of other host cell proteins, lipopolysaccharide endotoxin, and DNA in the recovered protein solution. The use of glycol ethers also allowed exploitation of temperature-change-induced phase splitting behavior to concentrate the desired product. Heating the aqueous EB extract solution to 55 degrees C formed two liquid phases: a glycol ether-rich phase and an aqueous product phase containing the great majority of the product protein. This phase-splitting step yielded an approximate 10-fold reduction in the volume of the initial product solution and a corresponding increase in the product's concentration.