Characterization of Bioactive Recombinant Human Lysozyme Expressed in Milk of Cloned Transgenic Cattle

BACKGROUND: There is great potential for using transgenic technology to improve the quality of cow milk and to produce biopharmaceuticals within the mammary gland. Lysozyme, a bactericidal protein that protects human infants from microbial infections, is highly expressed in human milk but is found in only trace amounts in cow milk. METHODOLOGY/PRINCIPAL FINDINGS: We have produced 17 healthy cloned cattle expressing recombinant human lysozyme using somatic cell nuclear transfer. In this study, we just focus on four transgenic cattle which were natural lactation. The expression level of the recombinant lysozyme was up to 25.96 mg/L, as measured by radioimmunoassay. Purified recombinant human lysozyme showed the same physicochemical properties, such as molecular mass and bacterial lysis, as its natural counterpart. Moreover, both recombinant and natural lysozyme had similar conditions for reactivity as well as for pH and temperature stability during in vitro simulations. The gross composition of transgenic and non-transgenic milk, including levels of lactose, total protein, total fat, and total solids were not found significant differences. CONCLUSIONS/SIGNIFICANCE: Thus, our study not only describes transgenic cattle whose milk offers the similar nutritional benefits as human milk but also reports techniques that could be further refined for production of active human lysozyme on a large scale

Recommended sleep duration is associated with higher consumption of fruits and vegetables; cross-sectional and prospective analyses from the UK Women’s Cohort Study

Background: High intakes of fruit and vegetable has been shown to protect against diseases and all-cause mortality however, the associations between sleep and fruit and vegetable consumption are not well characterized. This study aims to explore both cross-sectional and prospective associations between sleep duration and fruit and vegetable intakes in UK women. This is the first study to demonstrate the prospective association between sleep duration and fruit and vegetable consumption. Methods: Cross–sectional and prospective data were obtained from the UK Women’s Cohort Study. Sleep duration was assessed by self-report of average hours slept on weekdays and weekends and diet was assessed by a 4-day food diary at baseline and follow-up (~ 4 years later). Sleep duration was categorized as short (≤6 h/d), recommended (7–9 h/d) and long (≥9 h/d). Regression analyses adjusting for age, socio-economic status, smoking, ethnicity and total energy intake were used and restricted cubic spline models were developed to explore potential non-linear associations between sleep duration and fruit and vegetable intakes. Results: In adjusted cross-sectional analyses, short sleepers had on average 17 g/d (95% CI -30 to-4, p = 0.01) and long sleepers had 25 g/d (95% CI -39 to − 12, p < 0.001) less total fruits and vegetables compared to Recommended Sleepers (RS). In adjusted prospective analyses, short sleepers had on average 85 g/d (95% CI -144 to − 26, p = 0.005) less total fruits and vegetables in comparison to RS. Restricted cubic spline models showed that the cross-sectional (p < 0.001) and prospective (p = 0.001) associations between sleep duration and fruit and vegetable intakes were non-linear with women sleeping 7–9 h/d having the highest intakes. Conclusions: Fruit and vegetable consumption differed between sleep duration categories with UK women sleeping the recommended 7–9 h/day having the highest intake of fruits and vegetables in cross-sectional and prospective analyses. These findings suggest that sleeping the recommended duration is associated with higher consumption of fruits and vegetables. Sleep is an overlooked lifestyle factor in relation to fruit and vegetable consumption and more notice is vital. Further studies are required to clarify the underlying mechanisms for these associations

Development of an item bank for the EORTC Role Functioning Computer Adaptive Test (EORTC RF-CAT)

Background Role functioning (RF) as a core construct of health-related quality of life (HRQOL) comprises aspects of occupational and social roles relevant for patients in all treatment phases as well as for survivors. The objective of the current study was to improve its assessment by developing a computer-adaptive test (CAT) for RF. This was part of a larger project whose objective is to develop a CAT version of the EORTC QLQ-C30 which is one of the most widely used HRQOL instruments in oncology. Methods In accordance with EORTC guidelines, the development of the RF-CAT comprised four phases. Phase I involved the conceptualization of RF. In Phase II, a provisional list of items was defined and revised by experts in the field. In phase III, feedback was obtained from cancer patients in various countries. Phase IV comprised field testing in an international sample, calibration of the item bank, and evaluation of the psychometric performance of the RF-CAT. Results Phases I-III yielded a list of 12 items eligible for phase IV field-testing. The field-testing sample included 1,023 patients from Austria, Denmark, Italy, and the UK. Psychometric evaluation and item response theory analyses yielded 10 items with good psychometric properties. The resulting item bank exhibits excellent reliability (mean reliability = 0.85, median = 0.95). Using the RF-CAT may allow sample size savings from 11 % up to 50 % compared to using the QLQ-C30 RF scale. Conclusions The RF-CAT item bank improves the precision and efficiency with which RF can be assessed, promoting its integration into oncology research and clinical practice