Antibody degradation in tobacco plants: a predominantly apoplastic process.

BACKGROUND: Interest in using plants for production of recombinant proteins such as monoclonal antibodies is growing, but proteolytic degradation, leading to a loss of functionality and complications in downstream purification, is still a serious problem. RESULTS: In this study, we investigated the dynamics of the assembly and breakdown of a human IgG(1)κ antibody expressed in plants. Initial studies in a human IgG transgenic plant line suggested that IgG fragments were present prior to extraction. Indeed, when the proteolytic activity of non-transgenic Nicotiana tabacum leaf extracts was tested against a human IgG1 substrate, little activity was detectable in extraction buffers with pH > 5. Significant degradation was only observed when the plant extract was buffered below pH 5, but this proteolysis could be abrogated by addition of protease inhibitors. Pulse-chase analysis of IgG MAb transgenic plants also demonstrated that IgG assembly intermediates are present intracellularly and are not secreted, and indicates that the majority of proteolytic degradation occurs following secretion into the apoplastic space. CONCLUSIONS: The results provide evidence that proteolytic fragments derived from antibodies of the IgG subtype expressed in tobacco plants do not accumulate within the cell, and are instead likely to occur in the apoplastic space. Furthermore, any proteolytic activity due to the release of proteases from subcellular compartments during tissue disruption and extraction is not a major consideration under most commonly used extraction conditions

Orientation and experience in the perception of form: A study with the arizona whale-kangaroo

When subjects are presented with the Arizona whale-kangaroo, an ambiguous figure, perception of the whale is more common than perception of the kangaroo. However, this difference is smaller in Australian than American subjects. Perception of the kangaroo is more orientation dependent than perception of the whale, which is perceived at all orientations of the stimulus. Together with the difference between subject populations, this effect reveals an influence of past experience on the perception of this new ambiguous figure. Perception of the whale versus the kangaroo differs in both reconstrual of parts and realignment of the object-centered reference frame. Observers report reference frame reconstruals before reference frame reversals, shedding light on the organization of object memory

Relation of C-reactive protein to body fat distribution and features of the metabolic syndrome in Europeans and South Asians.

OBJECTIVE: To investigate the association between circulating C-reactive protein (CRP) concentrations and indices of body fat distribution and the insulin resistance syndrome in South Asians and Europeans. DESIGN: : Cross-sectional study. SUBJECTS: A total of 113 healthy South Asian and European men and women in West London (age 40-55 y, body mass index (BMI) 17-34 kg/m(2)). MEASUREMENTS: Fatness and fat distribution parameters (by anthropometry, dual-energy X-ray absorptiometry and abdominal CT scan); oral glucose tolerance test with insulin response; modified fat tolerance test; and CRP concentration by sensitive ELISA. RESULTS: Median CRP level in South Asian women was nearly double that in European women (1.35 vs 0.70 mg/1, P=0.05). Measures of obesity and CRP concentration were significantly associated in both ethnic groups. The correlation to CRP was especially strong among South Asians (P0.15). CONCLUSION: We suggest that adiposity and in particular visceral adipose tissue is a key promoter of low-grade chronic inflammation. This observation may in part account for the association of CRP with markers of the metabolic syndrome. Future studies should confirm whether CRP concentrations are elevated in South Asians and whether losing weight by exercise or diet, or reduction in visceral fat mass, is associated with reduction in plasma CRP concentrations

Radical scavenging activity of selected medicinal plants from Limpopo Province of South Africa

Plants collected from Limpopo province of South Africa were investigated for their antioxidative potential using the DPPH radical scavenging assay. Acetone extracts of Flueggea virosa had the highest antioxidant activity with an IC50 value of 30 ìg/ml, closely matching the ascorbic acid with an IC50 value of 25 ìg/ml. The lowest antioxidant readings were observed with extracts of Rhynchosia venulosa (root extract) and Ficus ingens (leaf extract). Acetone extract of Bridelia virosa leaves had the highest phenolic content (156 mg GAE/g extract), while the lowest content was recorded for R. venulosa root extract and leaf extract of F. ingens (8.3 and 17.7 mg GAE/g extract, respectively). There was a linear correlation between antioxidant activityand total phenolic content. Extracts with high phenolic content had low IC50 values, while extracts with low phenolic concentrations had high IC50 values

Breastfeeding Knowledge and Attitudes amongst HealthWorkers in a Health Care Facility in South-South Nigeria: The Need for Middle Level Health Manpower Development

This cross-sectional study was designed to assess the knowledge and attitudes of health workers in South-South Nigeria using a semi-structured, self-administered questionnaire completed by 36 respondents. Majority(69.4%) of participants were community health officers/ community health extension workers (CHO/CHEW). Findings indicate that human milk was acceptable to 97.2% of participating health workers. All participants approved of exclusive breastfeeding and giving colostrum to newborns.However, 8.3% identified < 4 months as the recommended length of exclusive breastfeeding duration, 8.3% specified > 6 months and 83.3% indicated 4–6 months. Only 36.1% knew that breastfeeding should last for a period of 2 years and beyond. One third of the respondents could not name more than two components of breast milk and 75.1% failedto identify more than three advantages of breastfeeding. Disturbingly, mental retardation (11.1%), diarrheal disease (2.8%) and failure to thrive (16.7%) were some of the disadvantages of breastfeeding identified. This study illustrates substantial and unacceptable knowledge gaps amongfrontline cadres of health workers. Immediate training and continuing medical education of health workers aimed at bridging these gaps, will help improve newborn health

Individualised benefit-harm balance of aspirin as primary prevention measure - a good proof-of-concept, but could have been better ...

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Aging Effects of Caenorhabditis elegans Ryanodine Receptor Variants Corresponding to Human Myopathic Mutations

Delaying the decline in skeletal muscle function will be critical to better maintenance of an active life-style in old age. The skeletal muscle ryanodine receptor, the major intracellular membrane channel through which calcium ions pass to elicit muscle contraction, is central to calcium ion balance, and is hypothesized to be a significant factor for age-related decline in muscle function. The nematode Caenorhabditis elegans is a key model system for the study of human aging and strains with modified C. elegans ryanodine receptors corresponding to human myopathic variants linked with malignant hyperthermia and related conditions were generated. The altered response of these strains to pharmacological agents reflected results of human diagnostic tests for individuals with these pathogenic variants. Involvement of nerve cells in the C. elegans responses may relate to rare medical symptoms concerning the central nervous system that have been associated with ryanodine receptor variants. These single amino acid modifications in C. elegans also conferred a reduction in lifespan and an accelerated decline in muscle integrity with age, supporting the significance of ryanodine receptor function for human aging

Rare earth elements (REEs) in the tropical South Atlantic and quantitative deconvolution of their non-conservative behavior

This study presents new concentration measurements of dissolved REEs (dREEs) along a full-depth east-west section across the tropical South Atlantic (~12°S), and uses these data to investigate the oceanic cycling of the REEs. Enrichment of dREEs, associated with the redox cycling of Fe-Mn oxides, is observed in the oxygen minimum zone (OMZ) off the African shelf. For deeper-waters, a multi-parameter mixing model was developed to deconvolve the relative importance of physical transport (i.e. water mass mixing) from biogeochemical controls on the dREE distribution in the deep Atlantic. This approach enables chemical processes involved in REE cycling, not apparent from the measurements alone, to be distinguished and quantified. Results show that the measured dREE concentrations below ~1000 m are dominantly controlled (>75%) by preformed REE concentrations resulting from water mass mixing. This result indicates that the linear correlation between dREEs and dissolved Si observed in Atlantic deep waters results from the dominantly conservative behaviour of these tracers, rather than from similar chemical processes influencing both dREEs and Si. Minor addition of dREEs (~10% of dNd and ~5% of dYb) is observed in the deep (>~4000 m) Brazil Basin, resulting from either remineralization of particles in-situ or along the flow path. Greater addition of dREEs (up to 25% for dNd and 20% for dYb) is found at ~1500 m and below ~4000 m in the Angola Basin near the African continental margin. Cerium anomalies suggest that different sources are responsible for these dREE addition plumes. The 1500 m excess is most likely attributed to dREE release from Fe oxides, whereas the 4000 m excess may be due to remineralization of calcite. Higher particulate fluxes and a more sluggish ocean circulation in the Angola Basin may explain why the dREE excesses in this basin are significantly higher than that observed in the Brazil Basin. Hydrothermal venting over the mid-Atlantic ridge acts as a regional net sink for light REEs, but has little influence on the net budget of heavy REEs. The combination of dense REE measurements with water mass deconvolution is shown to provide quantitative assessment of the relative roles of physical and biogeochemical processes in the oceanic cycling of REEs.X.-Y. Zheng was supported by the Clarendon Scholarship, the Exeter College Mandarin Scholarship from University of Oxford, the Chinese Student Awards from the Great Britain–China Educational Trust (GBCET) and W Wing Yip and Brothers bursaries.This is the author accepted manuscript. The final version is available from Elsevier via http://dx.doi.org/10.1016/j.gca.2016.01.01