Reference Protocol to Assess Analytical Performance of Higher Order Structural Analysis Measurements: Results from an Interlaboratory Comparison.

Measurements of protein higher order structure (HOS) provide important information on stability, potency, efficacy, immunogenicity, and biosimilarity of biopharmaceuticals, with a significant number of techniques and methods available to perform these measurements. The comparison of the analytical performance of HOS methods and the standardization of the results is, however, not a trivial task, due to the lack of reference protocols and reference measurement procedures. Here, we developed a protocol to structurally alter and compare samples of somatropin, a recombinant biotherapeutic, and describe the results obtained by using a number of techniques, methods and in different laboratories. This, with the final aim to provide tools and generate a pool of data to compare and benchmark analytical platforms and define method sensitivity to structural changes. Changes in somatropin HOS, induced by the presence of zinc at increasing concentrations, were observed, both globally and at more localized resolution, across many of the methods utilized in this study and with different sensitivities, suggesting the suitability of the protocol to improve understanding of inter- and cross-platform measurement comparability and assess analytical performance as appropriate.

Effects of carbohydrate dose and frequency on metabolism, gastrointestinal discomfort, and cross-country skiing performance.

This study investigated carbohydrate ingestion of varied doses and frequencies during a simulated cross-country skiing time trial. Ten men and three women (age: 30 ± 7 years; V ˙ O 2 m a x : 59.6 ± 5.7 mL/kg/min) completed four, 30-km classic technique roller skiing time trials on a treadmill. A 1:1 maltodextrin-fructose carbohydrate solution was provided at high (2.4 g/min; HC) and moderate (1.2 g/min; MC) ingestion rates, each at high (six feeds; HF) and low (two feeds; LF) frequencies. In the LF trials, blood glucose was elevated following carbohydrate ingestion (at 4 and 19 km) but was reduced at 14 and 29 km compared with HF strategies (P ≤ 0.05). Gastrointestinal discomfort was higher in HC-LF compared with all other trials (P ≤ 0.05). Whole-body lipid oxidation was lower and carbohydrate oxidation was higher in LF compared with HF trials (P ≤ 0.05). While performance time was not significantly different between trials (140:11 ± 15:31, 140:43 ± 17:40, 139:12 ± 15:32 and 140:33 ± 17:46 min:s in HC-HF, HC-LF, MC-HF, and MC-LF, respectively; P > 0.05), it was improved with trial order (P < 0.001). There was no effect of order on any other variable (P > 0.05). Altering carbohydrate dose or frequency does not affect cross-country ski performance. However, low-frequency carbohydrate ingestion resulted in poorer maintenance of euglycemia, reduced lipid oxidation, and increased gastrointestinal discomfort.

Lupus-Prone Mice Resist Immune Regulation and Transplant Tolerance Induction.

The strongly immunogenic environment in autoimmune diseases such as lupus may pose a stringent barrier to transplantation. Despite available murine models of lupus, transplant tolerance in this setting has yet to be fully investigated in highly penetrant genetic models of disease. Such studies are of clear clinical importance because lupus is a transplant indication in which transplanted kidneys have a substantially increased risk of rejection including a role for recurrent nephritis. In the fully penetrant B6.SLE123 mouse, we determined that CD4 T follicular helper and germinal center B cells were significantly expanded compared with healthy controls. We traced this expansion to resistance of effector CD4 T and B cells in B6.SLE123 mice to regulation by either CD4 T regulatory cells (CD4Tregs) or CD8 T regulatory cells (CD8Tregs), despite demonstrating normal function by Tregs in this strain. Finally, we determined that B6.SLE123 mice resist anti-CD45RB-mediated tolerance induction to foreign islet allografts, even in the absence of islet autoimmunity. Overall, B6.SLE123 lupus-prone mice are highly resistant to transplant tolerance induction, which provides a new model of failed tolerance in autoimmunity that may elucidate barriers to clinical transplantation in lupus through further cellular and genetic dissection.

A human flora-associated rat model of the breast-fed infant gut.

OBJECTIVES: Bacterial colonization of the infant gut may have important influences on the development of gastrointestinal, respiratory, and allergic disease. Early diet is a major determinant of the gut microflora. It is very difficult to carry out studies in human infants that can investigate the interaction of diet, flora, and mucosa. In this study we have developed an infant human flora-associated (IHFA) rat model to allow such investigation. METHODS: Germ-free infant rats were infected with fecal bacteria from exclusively breast-fed infants and were maintained on a modified infant formula for 8 weeks. The fecal and cecal contents were collected and compared with feces of breast-fed infants for bacterial populations, bacterial metabolites, and enzymes and for the ability to inhibit adhesion of pathogenic bacteria to human mucosal cells. RESULTS: The IHFA cecum and feces were dominated by lactic acid bacteria, Bifidobacterium, and lactobacilli, which were representative of the infant feces. The fecal short-chain fatty acid profile was dominated by acetic and lactic acid in a similar manner to human infant feces. Other bacterial metabolites were similar to those of the human infant. Rat intestinal samples were able to inhibit the adhesion of pathogens to mucosal cells, but to a lesser extent than the human samples. CONCLUSIONS: This IHFA infant model of the intestinal flora of the breast-fed infant is considered valid for studying the effect of diet on bacterial colonization and metabolism.

Climate change and forest fires.

This paper addresses the impacts of climate change on forest fires and describes how this, in turn, will impact on the forests of the United States. In addition to reviewing existing studies on climate change and forest fires we have used two transient general circulation models (GCMs), namely the Hadley Centre and the Canadian GCMs, to estimate fire season severity in the middle of the next century. Ratios of 2 x CO2 seasonal severity rating (SSR) over present day SSR were calculated for the means and maximums for North America. The results suggest that the SSR will increase by 10-50% over most of North America; although, there are regions of little change or where the SSR may decrease by the middle of the next century. Increased SSRs should translate into increased forest fire activity. Thus, forest fires could be viewed as an agent of change for US forests as the fire regime will respond rapidly to climate warming. This change in the fire regime has the potential to overshadow the direct effects of climate change on species distribution and migration.

The role of fire in the boreal carbon budget.

To reconcile observations of decomposition rates, carbon inventories, and net primary production (NPP), we estimated long-term averages for C exchange in boreal forests near Thompson, Manitoba. Soil drainage as defined by water table, moss cover, and permafrost dynamics, is the dominant control on direct fire emissions. In upland forests, an average of about 10-30% of annual NPP was likely consumed by fire over the past 6500 years since these landforms and ecosystems were established. This long-term, average fire emission is much larger than has been accounted for in global C cycle models and may forecast an increase in fire activity for this region. While over decadal to century times these boreal forests may be acting as slight net sinks for C from the atmosphere to land, periods of drought and severe fire activity may result in net sources of C from these systems.

Efficacy of self-care measures for perimenstrual syndrome (PMS).

More than 90% of women are affected by one or more of the signs and symptoms of perimenstrual syndrome (PMS), which causes disruptions at work and in family relations and can be economically costly. Self-care measures relating to lifestyle modifications, most particularly nutrition and exercise, can be helpful in preventing and/or alleviating the number and severity of symptoms. This quasi-experimental study of a pre-test and post-test design showed a statistically significant increase in self-care measures for PMS with a significant decrease in symptoms, thus impacting on the control of PMS.

The PIE system: complete planning and documentation of nursing care.

To address a number of difficulties with nursing documentation, a process-oriented documentation system called the Problem-Intervention-Evaluation (PIE) system was developed and implemented on a 35-bed medical unit at Craven County Hospital, New Bern, North Carolina. The PIE system simplifies the documentation process and unifies the care plan and progress notes into a complete, concise record of nursing care actually planned and provided. Informal evaluations indicate that the PIE system increases job satisfaction for nursing staff, improves the quality of documentation, and promotes continuity of care from shift to shift and from day to day. The article raises questions for further evaluation.