Relation between cell disruption conditions, cell debris particle size, and inclusion body release

The efficiency of physical separation of inclusion bodies from cell debris is related to cell debris size and inclusion body release and both factors should be taken into account when designing a process. In this work, cell disruption by enzymatic treatment with lysozyme and cellulase, by homogenization, and by homogenization with ammonia pretreatment is discussed. These disruption methods are compared on the basis of inclusion body release, operating costs, and cell debris particle size. The latter was measured with cumulative sedimentation analysis in combination with membrane-associated protein quantification by SDS-PAGE and a spectrophotometric pepticloglycan quantification method. Comparison of the results obtained with these two cell debris quantification methods shows that enzymatic treatment yields cell debris particles with varying chemical composition, while this is not the case with the other disruption methods that were investigated. Furthermore, the experiments show that ammonia pretreatment with homogenization increases inclusion body release compared to homogenization without pretreatment and that this pretreatment may be used to control the cell debris size to some extent. The enzymatic disruption process gives a higher product release than homogenization with or without ammonia pretreatment at lower operating costs, but it also yields a much smaller cell debris size than the other disruption process. This is unfavorable for centrifugal inclusion body purification in this case, where cell debris is the component going to the sediment and the inclusion body is the floating component. Nevertheless, calculations show that centrifugal separation of inclusion bodies from the enzymatically treated cells gives a high inclusion body yield and purity. (C) 2004 Wiley Periodicals, Inc.

Family average income and body mass index above the healthy weight range among urban and rural residents in regional Mainland China

Objective: To explore the relationship between family average income (FAI; an index of socio-economic status) and body mass index (BMI; a widely used, inexpensive indicator of weight status) above the healthy weight range in a region of Mainland China. Design: Population-based cross-sectional study, conducted between October 1999 and March 2000 on a sample of regular local residents aged 35 years or older who were selected by random cluster sampling. Setting: Forty-five administrative villages selected from three urban districts and two rural counties of Nanjing municipality, Mainland China, with a regional population of 5.6 million. Subjects: In total, 29 340 subjects participated; 67.7% from urban and 32.3% from rural areas; 49.8% male and 50.2% female. The response rate among eligible participants was 90.1%. Results: The proportion of participants classified as overweight was 30.5%, while 7.8% were identified as obese. After adjusting for possible confounding variables (age, gender, area of residence, educational level, occupational and leisure-time physical activity, daily vegetable consumption and frequency of red meat intake), urban participants were more likely to be overweight or obese relative to their rural counterparts, more women than men were obese, and participants in the lowest FAI tertile were the least likely to be above the healthy weight range. Conclusions: The proportion of adults with BMI above the healthy weight range was positively related to having a higher socio-economic status (indexed by FAI) in a regional Chinese population.

Multiple imputation for body mass index: lessons from the Australian Longitudinal Study on Women's Health

In large epidemiological studies missing data can be a problem, especially if information is sought on a sensitive topic or when a composite measure is calculated from several variables each affected by missing values. Multiple imputation is the method of choice for 'filling in' missing data based on associations among variables. Using an example about body mass index from the Australian Longitudinal Study on Women's Health, we identify a subset of variables that are particularly useful for imputing values for the target variables. Then we illustrate two uses of multiple imputation. The first is to examine and correct for bias when data are not missing completely at random. The second is to impute missing values for an important covariate; in this case omission from the imputation process of variables to be used in the analysis may introduce bias. We conclude with several recommendations for handling issues of missing data. Copyright (C) 2004 John Wiley Sons, Ltd.

Effect of body mass index changes between ages 5 and 14 on blood pressure at age 14 - Findings from a birth cohort study

Weight reduction in clinical populations of severely obese children has been shown to have beneficial effects on blood pressure, but little is known about the effect of weight gain among children in the general population. This study compares the mean blood pressure at 14 years of age with the change in overweight status between ages 5 and 14. Information from 2794 children born in Brisbane, Australia, and who were followed up since birth and had body mass index (BMI) and blood pressure measurements at ages 5 and 14 were used. Systolic and diastolic blood pressure at age 14 was the main outcomes and different patterns of change in BMI from age 5 to 14 were the main exposure. Those who changed from being overweight at age 5 to having normal BMI at age 14 had similar mean blood pressures to those who had a normal BMI at both time points: age- and sex-adjusted mean difference in systolic blood pressure 1.54 ( - 0.38, 3.45) mm Hg and in diastolic blood pressure 0.43 ( - 0.95, 1.81) mm Hg. In contrast, those who were overweight at both ages or who had a normal BMI at age 5 and were overweight at age 14 had higher blood pressure at age 14 than those who had a normal BMI at both times. These effects were independent of a range of potential confounding factors. Our findings suggest that programs that successfully result in children changing from overweight to normal-BMI status for their age may have important beneficial effects on subsequent blood pressure.

Body-mass management of Australian lightweight rowers prior to and during competition

Purpose: Although the body-mass management strategies of athletes in high-participation weight-category sports such as wrestling have been thoroughly investigated, little is known about such practices among lightweight rowers. This study examined the body-mass management practices of lightweight rowers before competition and compared these with current guidelines of the International Federation of Rowing Association (FISA). Quantification of nutrient intake in the 1-2 h between weigh-in and racing was also sought. Methods: Lightweight rowers (N = 100) competing in a national regatta completed a questionnaire that assessed body-mass management practices during the 4 wk before and throughout a regatta plus recovery strategies after weigh-in. Biochemical data were collected immediately after weigh-in to validate questionnaire responses. Responses were categorized according to gender and age category (Senior B or younger than 23 yr old, i.e., U23, Senior A or OPEN, i.e., open age limit) for competition. Results: Most athletes (male U23 76.5%, OPEN 92.3%; female U23 84.0%, OPEN 94.1%) decreased their body mass in the weeks before the regatta at rates compliant with FISA guidelines. Gradual dieting, fluid restriction, and increased training load were the most popular methods of body-mass management. Although the importance of recovery after weigh-in was recognized by athletes, nutrient intake and especially sodium (male U23 5.3 ± 4.9, OPEN 7.7 ± 5.9; female U23 5.7 ± 6.8, OPEN 10.2 ± 5.4 mg-kg(-1)) and fluid intake (male U23 12.1 ± 7.1, OPEN 13.5 ± 8.1; female U23 9.4 ± 7.4, OPEN 14.8 ± 6.9 mL.kg(-1)) were below current sports nutrition recommendations. Conclusion: Few rowers were natural lightweights; the majority reduced their body mass in the weeks before a regatta. Nutritional recovery strategies implemented by lightweight rowers after weigh-in were not consistent with current guidelines.

Impact of two different body mass management strategies on repeat rowing performance

Purpose: The present study was conducted to examine the impact of acute weight loss on repeat 2000-m rowing ergometer performance during a simulated multiday regatta. and to compare two different body mass management strategies between races. Methods: Competitive rowers (N = 16) were assigned to either a control (CON), partial recovery (RECpartial), or complete recovery (RECcomplete) group. Volunteers completed four trials, each separated by 48 h. No weight restrictions were imposed for the first trial. Thereafter, athletes in RECpartial and RECcomplete were required to reduce their body mass by 4% in the 24 h before trial 2, again reaching this body mass before the final two trials. No weight restrictions were imposed on CON. Aggressive nutritional recovery strategies were used in the 2 h following weigh-in for all athletes. These strategies were maintained for the 12-16 h following racing for RECcomplete with the aim of restoring at least three quarters of the original 4% body mass loss. Postrace recovery strategies were less aggressive in RECpartial; volunteers were encouraged to restore no more than half of their initial 4% body mass loss. Results: Acute weight loss increased time to complete the first at-weight performance trial by a small margin (mean 3.0, 95% CI -0.3 to 6.3 s, P = 0.07) when compared with the CON response. This effect decreased when sustained for several day,. Aggressive postrace recovery strategies tended to eliminate the effect of acute Weight loss on subsequent performance. Conclusion: Acute weight loss resulted in a small performance compromise that was reduced or eliminated when repeated over several days. Athletes should be encouraged to maximize recovery in the 12-16 h following racing when attempting to optimize subsequent performance.

The chicken ovalbumin upstream promoter-transcription factors modulate genes and pathways involved in skeletal muscle cell metabolism

The chicken ovalbumin upstream promoter-transcription factors ( COUP-TFs) are orphan members of the nuclear hormone receptor ( NR) superfamily. COUP-TFs are involved in organogenesis and neurogenesis. However, their role in skeletal muscle ( and other major mass tissues) and metabolism remains obscure. Skeletal muscle accounts for similar to 40% of total body mass and energy expenditure. Moreover, this peripheral tissue is a primary site of glucose and fatty acid utilization. We utilize small interfering RNA ( siRNA)-mediated attenuation of Coup-TfI and II ( mRNA and protein) in a skeletal muscle cell culture model to understand the regulatory role of Coup-Tfs in this energy demanding tissue. This targeted NR repression resulted in the significant attenuation of genes that regulate lipid mobilization and utilization ( including Ppar alpha, Fabp3, and Cpt-1). This was coupled to reduced fatty acid beta-oxidation. Additionally we observed significant attenuation of Ucp1, a gene involved in energy expenditure. Concordantly, we observed a 5-fold increase in ATP levels in cells with siRNA-mediated repression of Coup-TfI and II. Furthermore, the expression of classical liver X receptor ( LXR) target genes involved in reverse cholesterol transport ( Abca1 and Abcg1) were both significantly repressed. Moreover, we observed that repression of the Coup-Tfs ablated the activation of Abca1, and Abcg1 mRNA expression by the selective LXR agonist, T0901317. In concordance, Coup-Tf-siRNA-transfected cells were refractory to Lxr-mediated reduction of total intracellular cholesterol levels in contrast to the negative control cells. In agreement Lxr-mediated activation of the Abca1 promoter in Coup-Tf-siRNA cells was attenuated. Collectively, these data suggest a pivotal role for Coup-Tfs in the regulation of lipid utilization/cholesterol homeostasis in skeletal muscle cells and the modulation of Lxr-dependent gene regulation.

Change in lean body mass Is a major determinant of change in areal bone mineral density of the proximal femur: A 12-year observational study

Our objective was to assess the contribution of lean body mass (LBM) and fat body mass (FBM) to areal bone mineral density (aBMD) in women during the years surrounding menopause. We used a 12-year observational design. Participants included 75 Caucasian women who were premenopausal, 53 of whom were available for follow-up. There were two measurement periods: baseline and 12-year follow-up. At both measurement periods, bone mineral content and aBMD of the proximal femur, posterior-anterior lumbar spine, and total body was assessed using dual-energy X-ray absorptiometry (DXA). LBM and FBM were derived from the total-body scans. General health, including current menopausal status, hormone replace therapy use, medication use, and physical activity, was assessed by questionnaires. At the end of the study, 44% of the women were postmenopausal. After controlling for baseline aBMD, current menopausal status, and current hormone replacement therapy, we found that change in LBM was independently associated with change in aBMD of the proximal femur (P = 0.001). The cross-sectional analyses also indicated that LBM was a significant determinant of aBMD of all three DXA-scanned sites at both baseline and follow-up. These novel longitudinal data highlight the important contribution of LBM to the maintenance of proximal femur bone mass at a key time in women's life span, the years surrounding menopause.

An FTO variant is associated with Type 2 diabetes in South Asian populations after accounting for body mass index and waist circumference

Aims - A common variant, rs9939609, in the FTO (fat mass and obesity) gene is associated with adiposity in Europeans, explaining its relationship with diabetes. However, data are inconsistent in South Asians. Our aim was to investigate the association of the FTO rs9939609 variant with obesity, obesity-related traits and Type 2 diabetes in South Asian individuals, and to use meta-analyses to attempt to clarify to what extent BMI influences the association of FTO variants with diabetes in South Asians. Methods - We analysed rs9939609 in two studies of Pakistani individuals: 1666 adults aged = 40 years from the Karachi population-based Control of Blood Pressure and Risk Attenuation (COBRA) study and 2745 individuals of Punjabi ancestry who were part of a Type 2 diabetes case–control study (UK Asian Diabetes Study/Diabetes Genetics in Pakistan; UKADS/DGP). The main outcomes were BMI, waist circumference and diabetes. Regression analyses were performed to determine associations between FTO alleles and outcomes. Summary estimates were combined in a meta-analysis of 8091 South Asian individuals (3919 patients with Type 2 diabetes and 4172 control subjects), including those from two previous studies. Results - In the 4411 Pakistani individuals from this study, the age-, sex- and diabetes-adjusted association of FTO variant rs9939609 with BMI was 0.45 (95% CI 0.24–0.67) kg/m2 per A-allele (P = 3.0 × 10-5) and with waist circumference was 0.88 (95% CI 0.36–1.41) cm per A-allele (P = 0.001). The A-allele (30% frequency) was also significantly associated with Type 2 diabetes [per A-allele odds ratio (95% CI) 1.18 (1.07–1.30); P = 0.0009]. A meta-analysis of four South Asian studies with 8091 subjects showed that the FTO A-allele predisposes to Type 2 diabetes [1.22 (95% CI 1.14–1.31); P = 1.07 × 10-8] even after adjusting for BMI [1.18 (95% CI 1.10–1.27); P = 1.02 × 10-5] or waist circumference [1.18 (95% CI 1.10–1.27); P = 3.97 × 10-5]. Conclusions - The strong association between FTO genotype and BMI and waist circumference in South Asians is similar to that observed in Europeans. In contrast, the strong association of FTO genotype with diabetes is only partly accounted for by BMI.

Symbolic Dynamics in the Free-Fall Equal-Mass Three-Body Problem

The paper has been presented at the 12th International Conference on Applications of Computer Algebra, Varna, Bulgaria, June, 2006

The relationship of fear of intimacy, body image avoidance and body mass index

Issues of body image and ability to achieve intimacy are connected to body weight, yet remain largely unexplored and have not been evaluated by gender. The underlying purpose of this research was to determine if avoidant attitudes and perceptions of one's body may hold implications toward its use in intimate interactions, and if an above average body weight would tend to increase this avoidance. The National Health and Nutrition Examination Survey (NHANES, 1999-2002) finds that 64.5% of US adults are overweight, with 61.9% of women and 67.2% of men. The increasing prevalence of overweight and obesity in men and women shows no reverse trend, nor have prevention and treatment proven effective in the long term. The researcher gathered self-reported age, gender, height and weight data from 55 male and 58 female subjects (determined by a prospective power analysis with a desired medium effect size (r=.30) to determine body mass index (BMI), determining a mean age of 21.6 years and mean BMI of 25.6. Survey instruments consisted of two scales that are germane to the variables being examined. They were (1) Descutner and Thelen of the University of Missouri‘s (1991) Fear-of-Intimacy scale; and (2) Rosen, Srebnik, Saltzberg, and Wendt's (1991) Body Image Avoidance Questionnaire. Results indicated that as body mass index increases, fear of intimacy increases (p<0.05) and that as body mass index increases, body image avoidance increases (p<0.05). The relationship that as body image avoidance increases, fear of intimacy increases was not supported, but approached significance at (p<0.07). No differences in these relationships were determined between gender groups. For age, the only observed relationship was that of a difference between scores for age groups [18 to 22 (group 1) and ages 23 to 34 (group 2)] for the relationship of body image avoidance and fear of intimacy (p<0.02). The results suggest that the relationship of body image avoidance and fear of intimacy, as well as age, bear consideration toward the escalating prevalence of overweight and obesity. An integrative approach to body weight that addresses issues of body image and intimacy may prove effective in prevention and treatment.

Further evidence for the relationship between eating frequency, body mass index, and physical activity

The relationship between the frequency of eating, physical activity and Body Mass Index (BMI) was investigated. Seventy five women, aged 24 to 55, were recruited from Florida International University. Via interview, subjects provided information regarding demographics and habitual eating frequency over 24-hours, and completed both the Baecke Questionnaire of Habitual Physical Activity and the Health Insurance Plan of New York Questionnaire on Physical Activity. Pearson correlations and partial correlation coefficients were used to assess the relationship between eating frequency, physical activity, age, and BMI. Results revealed significant positive correlations between eating frequency and total physical activity scores, and leisure time physical activity scores, but not between eating frequency and physical activity on the job. Partial correlations suggest that there may be an effect of eating frequency on BMI both through an effect on physical activity and through another mechanism. These results suggest that more frequent eaters tend to be more physically active, which may partially explain why lower body weights is associated with more frequent eating.

Contrasting effects of summer and winter warming on body mass explain population dynamics in a food-limited Arctic herbivore

ACKNOWLEDGEMENTS We thank the Governor of Svalbard for permission to undertaker the research. We are especially grateful to Steve Coulson, and the logistical and technical staff at the University Centre in Svalbard (UNIS) for supporting the field campaigns. The data collection would not have been possible without the contribution of numerous field assistants, including veterinary students from the Norwegian School of Veterinary Science. Statistical advice was provided by Mark Brewer and David Elston, BioSS. The work was supported mainly by grants from U.K. Natural Environment Research Council the Norwegian Research Council, and the Macaulay Development Trust. Additional financial support has come from the Amundsen Foundation, Centre for Ecology and Hydrology, The Macaulay Institute, the NINA, UNIS, and the Norwegian School of Veterinary Science