Glycan Residues Balance Analysis - GReBA: A novel model for the N-linked glycosylation of IgG produced by CHO cells.

The structure of N-linked glycosylation is a very important quality attribute for therapeutic monoclonal antibodies. Different carbon sources in cell culture media, such as mannose and galactose, have been reported to have different influences on the glycosylation patterns. Accurate prediction and control of the glycosylation profile are important for the process development of mammalian cell cultures. In this study, a mathematical model, that we named Glycan Residues Balance Analysis (GReBA), was developed based on the concept of Elementary Flux Mode (EFM), and used to predict the glycosylation profile for steady state cell cultures. Experiments were carried out in pseudo-perfusion cultivation of antibody producing Chinese Hamster Ovary (CHO) cells with various concentrations and combinations of glucose, mannose and galactose. Cultivation of CHO cells with mannose or the combinations of mannose and galactose resulted in decreased lactate and ammonium production, and more matured glycosylation patterns compared to the cultures with glucose. Furthermore, the growth rate and IgG productivity were similar in all the conditions. When the cells were cultured with galactose alone, lactate was fed as well to be used as complementary carbon source, leading to cell growth rate and IgG productivity comparable to feeding the other sugars. The data of the glycoprofiles were used for training the model, and then to simulate the glycosylation changes with varying the concentrations of mannose and galactose. In this study we showed that the GReBA model had a good predictive capacity of the N-linked glycosylation. The GReBA can be used as a guidance for development of glycoprotein cultivation processes.

Business recovery: an assessment framework.

This paper presents a Business Recovery Assessment Framework (BRAF) to help researchers and practitioners design robust, repeatable, and comparable studies of business recovery in various post-disruption contexts. Studies assessing business recovery without adequately considering the research aims, recovery definitions, and indicators can produce misleading findings. The BRAF is composed of a series of steps that guide the decisions that researchers need to make to ensure: (i) that recovery is indeed being measured; (ii) that the indicators of recovery that are selected align with the objectives of the study and the definition of recovery; and, where necessary, (iii) that appropriate comparative control variables are in place. The paper draws on a large dataset of business surveys collected following the earthquakes in Canterbury, New Zealand, on 4 September 2010 and 22 February 2011 to demonstrate the varied conclusions that different recovery indicators can produce and to justify the need for a systematic approach to business recovery assessments.

Organisational resilience following the Darfield earthquake of 2010.

This paper presents the preliminary findings of a study on the resilience and recovery of organisations following the Darfield earthquake in New Zealand on 4 September 2010. Sampling included organisations proximal and distal to the fault trace, organisations located within central business districts, and organisations from seven diverse industry sectors. The research captured information on the challenges to, the impacts on, and the reflections of the organisations in the first months of recovery. Organisations in central business districts and in the hospitality sector were most likely to close, while organisations that had perishable stock and livestock were more heavily reliant on critical services. Staff well-being, cash flow, and customer loss were major concerns for organisations across all sectors. For all organisations, the most helpful factors in mitigating the effects of the earthquake were their relationship with staff members, the design and type of buildings, and critical service continuity or swift reinstatement of services.

Model-based design and control of a small-scale integrated continuous end-to-end mAb platform.

A continuous integrated bioprocess available from the earliest stages of process development allows for an easier, more efficient and faster development and characterization of an integrated process as well as production of small-scale drug candidates. The process presented in this article is a proof-of-concept of a continuous end-to-end monoclonal antibody production platform at a very small scale based on a 200 ml alternating tangential flow filtration perfusion bioreactor, integrated with the purification process with a model-based design and control. The downstream process, consisting of a periodic twin-column protein A capture, a virus inactivation, a CEX column and an AEX column, was compactly implemented in a single chromatography system, with a purification time of less than 4 hr. Monoclonal antibodies were produced for 17 days in a high cell density perfusion culture of CHO cells with titers up to 1.0 mg/ml. A digital twin of the downstream process was created by modelling all the chromatography steps. These models were used for real-time decision making by the implementation of control strategies to automatize and optimize the operation of the process. A consistent glycosylation pattern of the purified product was ensured by the steady state operation of the process. Regarding the removal of impurities, at least a 4-log reduction in the HCP levels was achieved. The recovery yield was up to 60%, and a maximum productivity of 0.8 mg/ml/day of purified product was obtained.

Combined effects of glycosylation precursors and lactate on the glycoprofile of IgG produced by CHO cells.

The glycosylation profile of therapeutic monoclonal antibodies (mAbs) is a crucial quality parameter for industrial Immunoglobulin G (IgG) production. Several alternative carbon sources, which function as glycosylation precursors, have been reported to impact the glycosylation pattern. Since the cells give priority to glucose uptake, the presence of this substrate can lower the effects of alternative sugars on the glycosylation. In order to get a better understanding of the influence of alternative sugars on the glycosylation and to investigate how they impact each other, combinations of mannose, fructose, galactose and fucose were fed to Chinese hamster ovary (CHO) cells in batch culture when the glucose became depleted and the lactate, accumulated in the culture, was used as carbon source. Feeding with a feed containing mannose or glucose decreased by 3-7% the percentage of high mannose glycans compared to a feed without mannose or glucose. Feeding with a feed containing galactose led to 8-20% increase of monogalactoglycans (G1) glycans and 2-6% rise of digalactoglycans (G2) glycans compared to feeding without galactose or glucose. The cells fed with fucose exhibited a significantly higher concentration of intracellular GDP-Fucose. This work indicates that a feeding strategy based on non-glucose sugars and potentially lactate, could be adopted to obtain a targeted glycosylation profile.

Alcohol use, misuse, abuse, and dependence in later adulthood.

Considerable research has focused on alcohol problems in older adults, but the clinical utilization of this knowledge has lagged at least 3 decades behind the scientific developments. This unfortunate situation takes on added significance as the "baby boomer" generation ages because more of them drink more often in larger quantities than previous generations. This chapter focuses more on the ramifications of use, misuse, and abuse than on chronic dependence because the prevalence in the former categories far outweighs the latter. Older alcohol misusers and abusers are at excess risk for myriad physical problems and premature death because alcohol interacts with the natural aging process in negative ways to increase risks for injuries, hypertension, cardiac dysrhythmic events, cancers, gastrointestinal problems, neurocognitive deficits, bone loss, and emotional challenges, most notably depression. Low volume and less than daily alcohol consumption appear to be protective against blood clots in the coronary and brain vessels, bone loss and falls, and cognitive decline compared with current abstainers. At higher levels, alcohol has the opposite effect. Research findings strongly support positive outcomes of case finding, referral, and treatment of older adults who are misusing or abusing alcohol. However, there is ample evidence that health care providers across the spectrum of primary, acute, and long-term care ignore the signs and symptoms of alcohol misuse and abuse in their older patients and treat symptoms and sequelae of the abuse rather than confronting the abuse itself. Recommendations for changes in practice are made together with ideas for additional research in several areas where the current state of knowledge is inadequate, conflicting, or based on narrowly homogeneous samples.

The case for alcohol research as a focus of study by nurse researchers.

Sixty percent of the U.S. population drinks alcohol. Although numerous investigators have shown that low-volume alcohol intake has positive influences on cardiovascular health, bone density, and cognition, there is a fine line between positive, neutral, and negative ramifications of alcohol consumption on health. Alcohol accounts for 7% of the global burden of disease and injury from all causes and for 10% to 11% of all illnesses and death each year worldwide. So alcohol use is a two-edged sword. Psychiatric nurses have a long history of involvement with alcoholic patients, and alcohol users, misusers, and abusers comprise a significant percentage of the patient load in every specialty and subspecialty of nursing. Yet nursing education has neglected this important area of content in general nursing curricula, staff development has not trained mainstream nurses to routinely assess for alcohol problems among hospital patients, and primary care providers have failed to do case finding. During the past 25 years, the federal government has funded curricular and faculty development programs to bring alcohol information into the core of health provider training. The process has been halting and minimally successful at best. One ramification of the inattention to alcohol-related education is the dearth of nurse-scientists engaged in programs of research in the field of alcohol abuse. A federally funded faculty development program in the 1990s focused attention on this issue, and a small cadre of nurses were trained to do related research. Several of the authors in this volume are members of that group. A brief overview of the focus of each of the remaining chapters in this volume is presented. A rationale is provided for the importance of this area of research for nursing knowledge and quality patient care in essentially all areas of nursing practice. Finally, several burning research questions are posed that would most appropriately be answered through nursing research.

A theoretical model of the role of brain stem nuclei in alcohol-mediated arrhythmogenesis in older adults.

Uncertainty about the mechanism of alcohol-mediated arrhythmogenesis and the effect of alcohol use on arrhythmic risk among older adults is an increasing concern in light of population aging and recent reports that moderate alcohol consumption may protect older adults against coronary artery disease. In this review, a theoretical model of the role of brain stem nuclei in alcohol-mediated arrhythmogenesis in older adults is developed. The model is based on the hypothesis that the effects of alcohol on central autonomic pathways of cardiac control may alter the threshold for alcohol-mediated arrhythmogenesis among older adults. Findings from multiple lines of research including cellular, electrophysiological, epidemiological, experimental, and clinical studies in human, animal, and in vitro models were synthesized in developing the model. Suggestions for future research on the topic of alcohol-mediated arrhythmogenesis in older adults are offered.

The association between moderate drinking and heart rate variability in healthy community-dwelling older women.

The relationships among moderate alcohol use, autonomic tone, and arrhythmogenesis in older adults have not been adequately studied. Knowledge about these relationships is of increasing importance in light of population aging and recent epidemiological findings that associate moderate alcohol use with decreased rates of coronary artery disease. The purpose of this study was to assess the association between moderate drinking and autonomic tone in older women. Fifty-two Caucasian female participants (age 69 +/- 5.2) were enrolled in the study. Autonomic tone was estimated by time-domain and frequency-domain measures of heart rate variability. Multivariate analysis revealed that alcohol consumption rate in the sample accounted for approximately one third of the 24-h variability in the SDNN and the SDANN, measures of variability cycle lengths of 24-h and more than 5-min, respectively. Significant contributions of alcohol consumption rate to the shorter-term time-domain measures rMSSD and ASDNN, all frequency-domain measures, and HR were not confirmed. However, repeated measures ANOVA revealed that, between the hours of 0000 and 0600, women who drank approximately 0.5 to 3 standard drinks per day had significantly lower [log] HF and [log] LF power compared to abstainers and a tendency toward sympathetic predominance during the evening and nighttime hours. The authors discuss the implications of these findings.

A review of the second decade of the Annual Review of Nursing Research series.

This chapter includes a review of the second decade of Annual Review of Nursing Research, Volumes 11-20. The authors analyze the content of these volumes and summarize the significant changes in the nursing scientific community. Also described are the contextual changes related to the development of nursing research.

Predictors of alcohol misuse and abuse in older women.

PURPOSE: To determine the predictive ability of self-report questions, physical measures, and biomarkers to detect alcohol misuse and abuse among older women. DESIGN AND METHODS: Healthy women volunteers age 60 and older who fit selection criteria were enrolled. The 135 participants were divided into nondrinkers (ND; n = 63) and drinkers (D; n = 72) based on self-reports of quantity and frequency of standard drinks consumed per month. The mean ages for the groups were 69.2 (ND) and 69.6 (D). FINDINGS: The best predictor was a score >0 on the T-ACE, a four-item instrument to detect alcohol abuse. Other significant predictors were: (a) behaviors: smoking, mixing over-the-counter (OTC) drugs with alcohol, heavy coffee drinking, using alcohol to sleep, and less sleep latency; and (b) biomarkers: higher mean corpuscular volume (MCV), hemoglobin (Hgb), hematocrit (Hct), and high-density lipoprotein cholesterol (HDL). The heaviest drinker subgroup had more physical stigmata, including broken blood vessels in nose and larger liver spans. CONCLUSIONS: The "best predictor model" showed that older women who were at risk for alcohol misuse or abuse had T-ACE scores of 1 or higher, used two or more OTC drugs regularly, drank large amounts of coffee, used alcohol to fall asleep, and had less sleep latency. Because positive T-ACE scores have high sensitivity and specificity for alcohol abuse, scores of 1 or greater should be addressed in clinical settings, e.g., referrals for more definitive diagnoses and relevant treatment.

Outcomes of a program to enhance exercise self-efficacy and improve fitness in Black and Hispanic college-age women.

A quasi-experimental design was used to test the outcomes of an exercise program directed towards Black and Hispanic college-age women. Forty-four women (36 Black, 7 Hispanic, and 1 Black/Hispanic) attended exercise classes three times per week for 16 weeks. At program completion, women were classified as either high attendees (n = 26) or low attendees (n = 18). Compared to low attendees, the high attendees had significantly higher exercise self-efficacy (p <.001), perceived benefits and barriers (p =.004), aerobic fitness, flexibility, muscle strength, and percentage of body fat (all p <.001). Daily activity levels improved significantly in the high attendance group following the program (p <.001) and at 8 weeks post-program completion (p =.01).