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The non-obese diabetic (NOD) mouse develops spontaneous type 1 diabetes, with some features of disease that are very similar to the human disease. However, a proportion of NOD mice are naturally-protected from developing diabetes, and currently studies characterising this cohort are very limited. Here, using both immunofluorescence and multi-parameter flow cytometry we focus on the pancreatic islet morphology and immune infiltrate observed in naturally-protected NOD mice. We show that naturally-protected NOD mice are characterised by an increased frequency of insulin-containing, smaller sized, pancreatic islets. Although mice remain diabetes free, florid immune infiltrate remains. However, this immune infiltrate is skewed towards a regulatory phenotype in both T and B-cell compartments. Pancreatic islets have an increased frequency of IL-10 producing B cells and associated cell surface markers. Resident memory CD69+CD8+ T cells show a significant shift towards reduced CD103 expression, while CD4+ T cells have increased FoxP3+CTLA4+ expression. These data indicate that naturally-protected NOD mice have a unique islet signature and provide new insight into regulatory mechanisms within pancreatic islets.

In the past few years, huge advances have been made in techniques to analyse cells at an individual level using RNA sequencing, and many of these have precipitated exciting discoveries in the immunology of type 1 diabetes (T1D). This review will cover the first papers to use scRNAseq to characterise human lymphocyte phenotypes in T1D in the peripheral blood, pancreatic lymph nodes and islets. These have revealed specific genes such as IL-32 that are differentially expressed in islet –specific T cells in T1D. scRNAseq has also revealed wider gene expression patterns that are involved in T1D and can predict its development even predating autoantibody production. Single cell sequencing of TCRs has revealed V genes and CDR3 motifs that are commonly used to target islet autoantigens, although truly public TCRs remain elusive. Little is known about BCR repertoires in T1D, but scRNAseq approaches have revealed that insulin binding BCRs commonly use specific J genes, share motifs between donors and frequently demonstrate poly-reactivity. This review will also summarise new developments in scRNAseq technology, the insights they have given into other diseases and how they could be leveraged to advance research in the type 1 diabetes field to identify novel biomarkers and targets for immunotherapy.

Decreasing renal glomerular filtration rate (GFR) in association with increasing blood cadmium levels was reported in epidemiological studies of general populations. Dependence of cadmium clearance on GFR has implications for interpreting causation in these studies. Associations between cadmium clearance and creatinine clearance, a metric of GFR, were evaluated in a sample of the U.S. population. Blood to urine cadmium clearance and serum creatinine clearance were estimated in approximately 6000 individuals included in the National Health and Nutrition Examination Survey (NHANES 2009-2016). Linear regression models explained approximately 45% of variance in cadmium clearance in adults, with 74% of the explained variance attributed to creatinine clearance and 25% explained by age. In adolescents (12-20 years), linear regression models explained 55% of variance in cadmium clearance with99% of the explained variance attributed to creatinine clearance. The models predicted that halving creatinine clearance would result in a 40% decrease in cadmium clearance and a 20% rise in blood cadmium. Dependence of cadmium clearance on GFR has implications for assigning causation to studies in which increasing blood cadmium levels have been associated with increasing risk of low GFR. Statistical associations between blood cadmium and low GFR, such as elevated odds ratios in upper percentile strata of populations, may be partially a consequence of lower cadmium clearance in association with low GFR that is reverse causation.

Blood lead (Pb) clearance (CbPb) and serum creatinine clearance (CsCr), a metric of glomerular filtration rate (GFR), were estimated in approximately 7,600 subjects from the NHANES (2009-2016). Median CbPb in adults was 0.04 L/day (5th-95th percentile range: 0.01-0.12). Linear regression models explained approximately 68% of variance in CbPb in adults, with98% of explained variance attributed to CsCr. These results provide an improved quantitative understanding of the possible effects of reverse causality in the interpretation of studies of associations between blood Pb and decrements in GFR.

Autobiographical memory consists of distinct memory types varying from highly abstract to episodic. Self trait knowledge, which is considered one of the more abstract types of autobiographical memory, is thought to rely on regions of the autobiographical memory neural network implicated in schema representation, including the medial prefrontal cortex, and critically, not the medial temporal lobes. The current case study introduces an individual who, as a consequence of bilateral posterior cerebral artery strokes, experienced extensive medial temporal lobe damage with sparing of the medial prefrontal cortex. Interestingly, in addition to severe retrograde and anterograde episodic and autobiographical fact amnesia, this individual’s self trait knowledge was impaired for his current and pre-morbid personality traits. Yet, further assessment revealed that this individual had preserved conceptual knowledge for personality traits, could reliably and accurately rate another person’s traits, and could access his own self-concept in a variety of ways. In addition to autobiographical memory loss, he demonstrated impairment on non-personal semantic memory tests, most notably on tests requiring retrieval of unique knowledge. This rare case of amnesia suggests a previously unreported role for the medial temporal lobes in personal trait knowledge, which we propose reflects the critical role of this neural region in the storage and retrieval of personal semantics that are experience-near, meaning autobiographical facts grounded in spatiotemporal contexts.

The non-obese diabetic (NOD) mouse develops spontaneous type 1 diabetes, with some features of disease that are very similar to the human disease. However, a proportion of NOD mice are naturally-protected from developing diabetes, and currently studies characterising this cohort are very limited. Here, using both immunofluorescence and multi-parameter flow cytometry we focus on the pancreatic islet morphology and immune infiltrate observed in naturally-protected NOD mice. We show that naturally-protected NOD mice are characterised by an increased frequency of insulin-containing, smaller sized, pancreatic islets. Although mice remain diabetes free, florid immune infiltrate remains. However, this immune infiltrate is skewed towards a regulatory phenotype in both T and B-cell compartments. Pancreatic islets have an increased frequency of IL-10 producing B cells and associated cell surface markers. Resident memory CD69+CD8+T cells show a significant shift towards reduced CD103 expression, while CD4+T cells have increased FoxP3+CTLA4+expression. These data indicate that naturally-protected NOD mice have a unique islet signature and provide new insight into regulatory mechanisms within pancreatic islets.

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Structural color is a pervasive natural phenomenon, caused by photonic nanostructures that refract light. Diverse organisms employ structural color to mediate ecological interactions and create specific optical effects such as iridescence. Despite its importance for living systems, the developmental, genetic, and evolutionary processes that generate structural color largely remain mysterious. Here, we focus on simple photonic structures, thin film reflectors, in the lower lamina of Junonia butterfly scales. We present multiple lines of evidence that the thickness of the lamina quantitatively controls lamina color, which is an important determinant of overall wing color, even when pigments are also present. First, in a lineage of buckeye butterflies artificially selected for blue wing color for 12 generations, a thicker lamina resulted in a color shift from brown to blue. A similar lamina thickness increase explains the appearance of blue scales in butterflies with mutations in the optix wing patterning gene. Finally, lamina thickness variation underlies the color diversity that distinguishes seasonal variants, sexes, and species throughout the genus Junonia. Thus, quantitatively tuning a single dimension of the existing scale architecture allows butterflies to evolve a broad spectrum of hues over both microevolutionary and macroevolutionary time frames. Because the lower lamina is an intrinsic component of typical butterfly scales, our findings imply that lamina structural color influences wing color in most butterflies. Significance Statement Structural colors, which result from photonic nanostructures that refract light and can create iridescence, are an important tool for many organisms. We use thin films, which are morphologically simple nanostructures that generate structural color in the lower lamina of butterfly scales, to dissect how photonic structures evolve. By combining interspecies comparisons with two different experimental approaches—artificial selection on wing color, and genetically engineered mutation of the optix wing patterning gene—we demonstrate that lamina thickness controls the wavelength (hue) of the structural color. These lamina structural colors are ubiquitous in the genus Junonia, and determine wing color along with pigments. Our results suggest that lamina structural colors probably exist in most butterflies, and that tuning lamina thickness facilitates wing color evolution.

Sleep loss contributes to the development of cardiovascular, metabolic, and neurological disorders by promoting a systemic proinflammatory phenotype. The neuroendocrine-immune mechanisms contributing to such pathologies are poorly understood. The sympathetic nervous system (SNS) regulates immunity and is often activated following sleep disturbances. The aims of this study were to determine 1) the effect of SNS inhibition on inflammatory responses to sleep fragmentation (SF) and 2) whether homeostasis can be restored after 1 wk of recovery sleep. We measured stress responses (norepinephrine and corticosterone), gene expression levels of pro- and anti-inflammatory cytokines in peripheral (heart, liver, and spleen) tissues, and protein levels of cytokines and chemokines in serum of female mice that were subjected to acute SF for 24 h, chronic SF for 8 wk, or 7 days of recovery after chronic SF. In each experiment, SF and control mice were chemically sympathectomized with 6-hydroxydopamine (6-OHDA) or injected with vehicle. Both acute and chronic SF elevated mRNA and protein levels of cytokines in peripheral tissues. Changes in inflammatory responses mirrored stress-axes activation, with increased corticosterone and norepinephrine in SF mice. 6-OHDA treatment significantly alleviated SF-induced inflammation, thus providing evidence of SNS regulation of peripheral inflammation from SF. Effects of chronic SF were more severe than acute SF, and 1 wk of recovery from SF sufficiently alleviated peripheral inflammatory responses but not NE responses.

There are now a number of different mouse models for type 1 diabetes. The best known is the nonobese diabetic (NOD) mouse which has a genetic susceptibility to autoimmune diabetes with some features that are similar to human type 1 diabetes. The mice also have a propensity to other autoimmune diatheses, including autoimmune thyroid disease and sialadenitis. In addition, it is well known that environmental factors affect the incidence of disease in these mice. While there are other rodent models, including numerous transgenic and knockout models, as well as those that express human proteins, none of these develop spontaneous diabetes over a period of time, when the natural history can be studied. We focus here on the unmanipulated NOD mouse and discuss features of the husbandry and investigation of the mice that allow for use of these long-studied mice in the pathogenesis of an autoimmune type of diabetes.

Type 1 diabetes is an autoimmune disease resulting in the loss of insulin production and, consequently, hyperglycemia. The nonobese diabetic (NOD) mouse develops spontaneous diabetes with considerable similarity to the disease in humans. Immunological studies using the NOD mouse model allow for the investigation of the natural history of the disease and leukocyte and lymphocyte pathogenic and regulatory functions, as well as testing potential therapies for intervention. The analyses of the cellular events leading up to diabetes may utilize different in vitro cellular assays, immunohistochemistry, and in vivo adoptive transfer, to study mechanisms of the disease and the effects of therapeutic intervention. In this chapter, we describe some common techniques for phenotyping and mechanistic analyses of function, particularly of CD8+ T cells.

NOD mice, a model strain for human type 1 diabetes, express proinsulin (PI) in the thymus. However, insulin-reactive T cells escape negative selection, and subsequent activation of the CD8(+) T-cell clonotype G9C8, which recognizes insulin B15-23 via an αβ T-cell receptor (TCR) incorporating TRAV8-1/TRAJ9 and TRBV19/TRBJ2-3 gene rearrangements, contributes to the development of diabetes. In this study, we used fixed TRAV8-1/TRAJ9 TCRα-chain transgenic mice to assess the impact of PI isoform expression on the insulin-reactive CD8(+) T-cell repertoire. The key findings were: 1) PI2 deficiency increases the frequency of insulin B15-23-reactive TRBV19(+)CD8(+) T cells and causes diabetes; 2) insulin B15-23-reactive TRBV19(+)CD8(+) T cells are more abundant in the pancreatic lymph nodes of mice lacking PI1 and/or PI2; 3) overexpression of PI2 decreases TRBV19 usage in the global CD8(+) T-cell compartment; 4) a biased repertoire of insulin-reactive CD8(+) T cells emerges in the periphery regardless of antigen exposure; and 5) low-avidity insulin-reactive CD8(+) T cells are less affected by antigen exposure in the thymus than in the periphery. These findings inform our understanding of the diabetogenic process and reveal new avenues for therapeutic exploitation in type 1 diabetes.

BACKGROUND: Afterschool interventions have been found to improve the nutritional quality of snacks served. However, there is limited evidence on how these interventions affect children’s snacking behaviors. OBJECTIVE: To determine the impact of an afterschool intervention focused at the school district, site, family, and child levels on dietary consumption of foods and beverages served at snack. DESIGN: Secondary analysis of a group-randomized controlled trial. PARTICIPANTS/SETTING: Data were collected from 400 children at 20 Boston, MA afterschool sites before (fall 2010) and after (spring 2011) intervention implementation. INTERVENTION: The Out-of-School Nutrition and Physical Activity intervention aimed to promote fruits, vegetables, whole grains, and water, while limiting sugary drinks and trans fats. Researchers worked with district foodservice staff to change snack foods and beverages. Teams of afterschool staff participated in three three-hour learning collaborative sessions to build skills and created action plans for changing site practices. The intervention included family and child nutrition education. MAIN OUTCOME MEASURES: Research assistants observed dietary snack consumption using a validated measure on two days per site at baseline and follow-up. STATISTICAL ANALYSES PERFORMED: This study used multivariable regression models, accounting for clustering of observations, to assess the intervention effect, and conducted post-hoc stratified analyses by foodservice type. RESULTS: Children in intervention sites had greater decreases in consumption of juice (−0.61 ounces/snack, 95% CI −1.11 – −0.12), beverage calories (−29.0 kcal/snack, 95% CI −40.1 – −17.9), foods with trans fats (−0.12 servings/snack, 95% CI −0.19 – −0.04), total calories (−47.8 kcal/snack, 95% CI −68.3 – −27.3) and increases in consumption of whole grains (0.10 servings/snack, 95% CI 0.02–0.18) compared to controls. In post hoc analyses, sites with on-site foodservice had significant improvements for all outcomes (p

Background Providing drinking water to U.S. children during school meals is a recommended health promotion strategy and part of national nutrition policy. Urban school systems have struggled with providing drinking water to children, and little is known about how to ensure that water is served, particularly in afterschool settings. Purpose To assess the effectiveness of an intervention designed to promote water as the beverage of choice in afterschool programs. Design The Out of School Nutrition and Physical Activity Initiative (OSNAP) used a community-based collaboration and low-cost strategies to provide water after school. A group RCT was used to evaluate the intervention. Data were collected in 2010–2011 and analyzed in 2011. Setting/participants Twenty afterschool programs in Boston were randomized to intervention or control (delayed intervention). Intervention Intervention sites participated in learning collaboratives focused on policy and environmental changes to increase healthy eating, drinking, and physical activity opportunities during afterschool time (materials available at www.osnap.org). Collaboration between Boston Public Schools Food and Nutrition Services, afterschool staff, and researchers established water-delivery systems to ensure children were served water during snack time. Main outcome measures Average ounces of water served to children per day was recorded by direct observation at each program at baseline and 6-month follow-up over 5 consecutive school days. Secondary measures directly observed included ounces of other beverages served, other snack components, and water-delivery system. Results Participation in the intervention was associated with an increased average volume of water served (+3.6 ounces/day; p =0.01) during snack. On average, the intervention led to a daily decrease of 60.9 kcals from beverages served during snack ( p =0.03). Conclusions This study indicates the OSNAP intervention, including strategies to overcome structural barriers and collaboration with key actors, can increase offerings of water during afterschool snack. OSNAP appears to be an effective strategy to provide water in afterschool settings that can be helpful in implementing new U.S. Department of Agriculture guidelines regarding water availability during lunch and afterschool snack. Trial registration This study is registered at clinicaltrials.gov NCT01396473.

OBJECTIVE The role of reactive oxygen species (ROS) and their dissipation in type 1 diabetes pathogenesis have garnered considerable controversy. Our recent work has demonstrated the importance of NADPH oxidase (NOX) activity for type 1 diabetes development and modulating T-cell autoreactivity. We previously linked decreased monocyte ROS with diabetes resistance in the alloxan-resistant mouse, and NOD-Ncf1m1J mice with a genetic ablation of NOX activity had reduced and delayed type 1 diabetes compared with NOD mice. RESEARCH DESIGN AND METHODS To determine the required cellular sources of ROS that are necessary for type 1 diabetes initiation, we used antibody depletion and adoptive transfer experiments into NOD and NOD-Scid females, respectively. After receiving treatment, female mice were monitored for hyperglycemia and overt diabetes. RESULTS Depletion of macrophages and neutrophils fully protected NOD mice from type 1 diabetes. However, elimination of neutrophils alone showed no significant reduction or delay. Type 1 diabetes induction in NOD-Scid mice by adoptive transfer with NOD-Ncf1m1J splenocytes was significantly delayed compared with NOD splenocytes, suggesting macrophage ROS and modulation of effector responses are critical for diabetes. The adaptive immune response was also altered by the absence of NOX activity, as purified T cells from NOD-Ncf1m1J mice exhibited delayed transfer kinetics. Cotransfer experiments demonstrated the defect was intrinsic to NOX-deficient CD8+ T cells. After stimulation, cytotoxic T cells exhibited decreased effector function in the absence of superoxide production. CONCLUSIONS These data demonstrate that the impaired autoreactive response of NOX-deficient NOD-Ncf1m1J immune system results from an alteration in the antigen-presenting cell–T-cell axis rather than failure of neutrophils to act as effector cells and that ROS signaling is important for the initiation of β-cell–directed autoimmunity by T cells.

In 1922, Leonard Thompson received the first injections of insulin prepared from the pancreas of canine test subjects. From pancreatectomized dogs to the more recent development of animal models that spontaneously develop autoimmune syndromes, animal models have played a meaningful role in furthering diabetes research. Of these animals, the nonobese diabetic (NOD) mouse is the most widely used for research in type 1 diabetes (T1D) because the NOD shares several genetic and immunologic traits with the human form of the disease. In this article, the authors discuss the similarities and differences in NOD and human T1D and the potential role of NOD mice in future preclinical studies, aiming to provide a better understanding of the genetic and immune defects that lead to T1D.

The annual number of total hip and knee replacements roughly doubled between 1994 and 2004, and this trend is projected to continue. These surgical patients are often older and many have special postsurgical care and rehabilitation needs. However, historically they have been incorporated into the general orthopaedic patient population. The development of centers of excellence in joint replacement has become one method for meeting the special needs of this population. This article describes the journey one midwestern healthcare system has taken in establishing a consolidated, specialized joint replacement center, which has received the Joint Commission's Disease-Specific Care Certification for hip and knee replacement. Specific steps in the development of the center, implementation, evaluation and outcomes, and lessons learned are described.

Importance Millions of children attend after-school programs in the United States. Increasing physical activity levels of program participants could have a broad effect on children’s health. Objective To test the effectiveness of the Out of School Nutrition and Physical Activity (OSNAP) Initiative in increasing children’s physical activity levels in existing after-school programs. Design, Setting, and Participants Cluster-randomized controlled trial with matched program pairs. Baseline data were collected September 27 through November 12, 2010, with follow-up data collected April 25 through May 27, 2011. The dates of our analysis were March 11, 2014, through August 18, 2015. The setting was 20 after-school programs in Boston, Massachusetts. All children 5 to 12 years old in participating programs were eligible for study inclusion. Interventions Ten programs participated in a series of three 3-hour learning collaborative workshops, with additional optional opportunities for training and technical assistance. Main Outcomes and Measures Change in number of minutes and bouts of moderate to vigorous physical activity, vigorous physical activity, and sedentary activity and change in total accelerometer counts between baseline and follow-up. Results Participants with complete data were 402 racially/ethnically diverse children, with a mean age of 7.7 years. Change in the duration of physical activity opportunities offered to children during program time did not differ between conditions (−1.2 minutes; 95% CI, −14.2 to 12.4 minutes; P = .87). Change in moderate to vigorous physical activity minutes accumulated by children during program time did not differ significantly by intervention status (−1.0; 95% CI, −3.3 to 1.3; P = .40). Total minutes per day of vigorous physical activity (3.2; 95% CI, 1.8-4.7; P P P = .01) increased significantly during program time among intervention participants compared with control participants. Conclusions and Relevance Although programs participating in the OSNAP Initiative did not allot significantly more time for physical activity, they successfully made existing time more vigorously active for children receiving the intervention. Trial Registration clinicaltrials.gov Identifier:NCT01396473

We showed previously that hepatic vitamin A concentrations of captive rhesus monkeys (Macaca mulatta) are subtoxic to toxic, with livers exhibiting stellate cell hypertrophy and hyperplasia. Although marmoset (Callithrix jacchus) livers are also high in vitamin A, no stellate cell irregularities were observed. To further characterize the effects of high dietary vitamin A from preformed sources, stored serum samples were analyzed from monkeys used for biomedical research and housed at the Wisconsin Primate Research Center. The monkeys had been fed commercially available monkey diets, providing vitamin A (as retinyl acetate) at levels exceeding NRC recommendations by a factor of four. The serum from both rhesus and marmoset monkeys had total serum vitamin A (retinol, retinyl esters and metabolites) within the expected range for both species, i.e., 1.44 +/- 0.34 and 1.41 +/- 0.72 micromol/L serum for rhesus and marmoset monkeys, respectively. However, high serum retinyl ester concentrations as a percentage of total serum vitamin A were present in both species, 12 +/- 5.1% (range, 5.5-23%) for rhesus and 27 +/- 14% (range, 10-57%) for the marmosets. Serum retinol concentrations were normal, i.e., 1.21 +/- 0.28 (rhesus) and 0.92 +/- 0.43 micromol/L (marmoset), compared with published values.

Olfaction plays an important role in the foraging behavior of rodents, but little is known about how this sense varies among taxa that evolved in different ecological settings. Field experiments were conducted in 10- by 10-m enclosures to test for interspecific differences in foraging success for ponderosa pine (Pinus ponderosa) or sunflower seeds, based on olfaction, of 4 rodent species: 2 heteromyid rodents (Panamint kangaroo rat, Dipodomys panamintinus, and Great Basin pocket mouse, Perognathus parvus) and 2 nonheteromyid rodents (yellow pine chipmunk, Tamias amoenus, and deer mouse, Peromyscus maniculatus). All species found large, shallow caches more frequently than small, deep caches. Chipmunks had the lowest foraging success for buried sunflower seeds under dry conditions. Deer mice did as well as the heteromyid rodents on large shallow caches but found relatively few small, deep caches. The heteromyids were the only species that found small, deep sunflower seed caches under dry conditions, and Panamint kangaroo rats always found more caches than did Great Basin pocket mice. These results confirm that olfaction plays an important role in foraging of rodents for buried seeds and support the notion that the olfactory sense of rodent species has evolved greater sensitivity to seeds in different ecological contexts.

An analysis of epidemiological studies of associations between exposure to cadmium and kidney toxicity was conducted. Dose-response functions relating low-molecular-weight (LMW) proteinuria to various indices of cadmium dose (dietary cadmium intake, urinary cadmium excretion, or tissue cadmium burden) were obtained from 15 studies of diverse exposures (occupational, general environmental, environmental contamination). Estimates of the dose corresponding to probabilities of LMW proteinuria of 0.1, 0.15, or 0.2 were transformed from the reported dose units into corresponding estimates of target organ dose (microgram Cd/g renal cortex, RC) by simulation using a pharmacokinetics (PK) model. The median RC associated with a 0.1 probability (RC10M) of LMW proteinuria was predicted to be 153 micrograms Cd/g cortex (95% confidence interval [CI]: 84-263). The lower confidence limit on the RC10M (RC10L, 84 micrograms/g cortex) was predicted to be attained with a constant chronic intake of 1 microgram/kg/d in females or 2.2 micrograms/kg/d in males. The RC10L was 2.5-5 times higher than the median RCs predicted to result from dietary cadmium intake in U.S. nonsmokers (microgram Cd/g cortex: 33, females; 17, males) and 1.6-3 times higher than the corresponding 95th percentile RCs (53, females; 27, males). Additional exposure from smoking cigarettes (approximately 20 cigarettes/d, 3 micrograms Cd inhaled/d) was predicted to increase the median RC (microgram/g cortex) by approximately 45-70% (48, females; 29, males); however, predicted 95th percentile RCs for smokers (66, females; 38, males) were lower than the RC10L. These results indicate that, for most of the U.S. population, dietary-derived risks are likely to be negligible, in the absence of exposures from other sources.

BACKGROUND Existing approaches for measuring hemostasis parameters require multiple platforms, can take hours to provide results, and generally require 1–25 mL of sample. We developed a diagnostic platform that allows comprehensive assessment of hemostatic parameters on a single instrument and provides results within 15 min using 0.04 mL of blood with minimal sample handling. METHODS T2 magnetic resonance (T2MR) was used to directly measure integrated reactions in whole blood samples by resolving multiple water relaxation times from distinct sample microenvironments. Clotting, clot contraction, and fibrinolysis stimulated by thrombin or tissue plasminogen activator, respectively, were measured. T2MR signals of clotting samples were compared with images produced by scanning electron microscopy and with standard reference methods for the following parameters: hematocrit, prothrombin time, clot strength, and platelet activity. RESULTS Application of T2MR methodology revealed conditions under which a unique T2MR signature appeared that corresponded with the formation of polyhedral erythrocytes, the dynamics and morphology of which are dependent on thrombin, fibrinogen, hematocrit, and platelet levels. We also showed that the T2MR platform can be used for precise and accurate measurements of hematocrit (%CV, 4.8%, R2 = 0.95), clotting time (%CV, 3.5%, R2 = 0.94), clot strength (R2 = 0.95), and platelet function (93% agreement with light transmission aggregometry). CONCLUSIONS This proof-of-concept study demonstrates that T2MR has the potential to provide rapid and sensitive identification of patients at risk for thrombosis or bleeding and to identify new biomarkers and therapeutic targets with a single, simple-to-employ analytic approach that may be suitable for routine use in both research and diverse clinical settings.

Neutrophils are essential for successful host eradication of bacterial pathogens and for survival to polymicrobial sepsis. During inflammation, the bone marrow provides a large reserve of neutrophils that are released into the peripheral circulation where they traverse to sites of infection. Although neutrophils are essential for survival, few studies have investigated the mechanisms responsible for neutrophil mobilization from the bone marrow during polymicrobial sepsis. Using a cecal ligation and puncture model of polymicrobial sepsis, we demonstrated that neutrophil mobilization from the bone marrow is not dependent on TLR4, MyD88, TRIF, IFNARα/β, or CXCR2 pathway signaling during sepsis. In contrast, we observed that bone marrow CXCL12 mRNA abundance and specific CXCL12 levels are sharply reduced, whereas splenic CXCR4 mRNA and cell surface expression are increased during sepsis. Blocking CXCL12 activity significantly reduced blood neutrophilia by inhibiting bone marrow release of granulocytes during sepsis. However, CXCL12 inhibition had no impact on the expansion of bone marrow neutrophil precursors and hematopoietic progenitors. Bone marrow neutrophil retention by CXCL12 blockade prevented blood neutrophilia, inhibited peritoneal neutrophil accumulation, allowed significant peritoneal bacterial invasion, and increased polymicrobial sepsis mortality. We concluded that changes in the pattern of CXCL12 signaling during sepsis are essential for neutrophil bone marrow mobilization and host survival but have little impact on bone marrow granulopoiesis.

Computer-assisted insulin protocols (CAIPs) contain complex mathematical algorithms to assist with insulin dosing. This study compared the quality of glucose control utilizing a CAIP with a paper-based insulin protocol (PBIP).This before-after study identified consecutive patients who received continuous insulin therapy for at least 24 h. Patients were stratified into two groups (PBIP and CAIP). The target blood glucose range for both was 80-110 mg/dL. Hypoglycemia was defined as the percentage of patients with any glucose value40 mg/dL. Variability was measured by reporting the SD for each patients mean glucose value.There were 192 patients evaluated (PBIP, n = 145; CAIP, n = 47). More glucose readings were within target range using the CAIP protocol (49 ± 14% vs. 40 ± 12%, P0.001), but no difference in mean glucose was noted (113 ± 11 mg/dL with CAIP vs. 116 ± 11 mg/dL with PBIP, P = 0.067). The incidence of hypoglycemia was similar between the CAIP and PBIP groups, respectively (2.1% vs. 4.1%, P = 0.518). Glucose variability was lower with the CAIP (25 ± 9 mg/dL vs. 31 ± 11 mg/dL, P = 0.001). The CAIP required more frequent blood glucose assessments (16 ± 2 vs. 12 ± 2 per day, P0.001), more insulin dosing adjustments (14 ± 3 vs. 5 ± 2 per day, P0.001), and more time per day (84 ± 15 vs. 51 ± 8 min per patient, P0.001) compared with the PBIP.A CAIP will lead to minor improvements in glucose control and decrease glucose variability but will not change the rate of hypoglycemia or response to insulin therapy. These differences could largely be due to more aggressive monitoring and titrations required by a CAIP.

Sepsis, the systemic inflammatory response to microbial infection, induces changes in both innate and adaptive immunity that presumably lead to increased susceptibility to secondary infections, multiorgan failure, and death. Using a model of murine polymicrobial sepsis whose severity approximates human sepsis, we examined outcomes and defined requirements for survival after secondary Pseudomonas aeruginosa pneumonia or disseminated Listeria monocytogenes infection. We demonstrate that early after sepsis neutrophil numbers and function are decreased, whereas monocyte recruitment through the CCR2/MCP-1 pathway and function are enhanced. Consequently, lethality to Pseudomonas pneumonia is increased early but not late after induction of sepsis. In contrast, lethality to listeriosis, whose eradication is dependent upon monocyte/macrophage phagocytosis, is actually decreased both early and late after sepsis. Adaptive immunity plays little role in these secondary infectious responses. This study demonstrates that sepsis promotes selective early, impaired innate immune responses, primarily in neutrophils, that lead to a pathogen-specific, increased susceptibility to secondary infections.

Reactive oxygen species are used by the immune system to eliminate infections; however, they may also serve as signaling intermediates to coordinate the efforts of the innate and adaptive immune systems. In this study, we show that by eliminating macrophage and T cell superoxide production through the NADPH oxidase (NOX), T cell polarization was altered. After stimulation with immobilized anti-CD3 and anti-CD28 or priming recall, T cells from NOX-deficient mice exhibited a skewed Th17 phenotype, whereas NOX-intact cells produced cytokines indicative of a Th1 response. These findings were corroborated in vivo by studying two different autoimmune diseases mediated by Th17 or Th1 pathogenic T cell responses. NOX-deficient NOD mice were Th17 prone with a concomitant susceptibility to experimental allergic encephalomyelitis and significant protection against type 1 diabetes. These data validate the role of superoxide in shaping Th responses and as a signaling intermediate to modulate Th17 and Th1 T cell responses.

Protection of pancreatic beta cells is an approach to prevent autoimmune type 1 diabetes (T1D) and to protect transplanted islets. Reactive oxygen species (ROS) are important mediators of beta cell death during the development of T1D. We have examined the role of elevated ROS dissipation in the prevention of T1D using the ALR mouse strain. The selection of ALR, for resistance against alloxan-induced free radical-mediated diabetes, led to a strain of mice with an elevated systemic as well as pancreatic ROS dissipation. Independent genetic mapping studies have identified ALR-derived diabetes protective loci. Conplastic and congenic mouse as well as cell line studies have confirmed the genetic mapping and demonstrated that the elevated ROS dissipation protects ALR beta cells from autoimmune destruction. Our data support the hypothesis that elevated ROS dissipation protects beta cells against autoimmune destruction and prevents T1D development.

We investigated the role of cerebral motor structures during mental hand rotation. Neural activity was measured with event-related potentials (ERPs) in 16 healthy participants while they performed handedness judgments of visually presented hands. Mental rotation was associated with ERP amplitude modulations as early as 170 ms but most strongly during a time window of about 600–800 ms. Source analysis of ERPs during these time windows indicated generators in bilateral extrastriate and parietal cortices. The results do not support a direct involvement of anterior motor cortices in the neural computations underlying mental rotation. However, motor regions may play a role in providing ongoing kinaesthetic feedback during mental rotation or in checking the results of the imagined transformation.

We measured brain potentials from human subjects performing a mental rotation task requiring right-left judgments of misoriented hands, and a control task requiring palm-back judgments of the same stimuli. High-density, 128-channel event-related potentials (ERPs) were recorded from 16 normal, right-handed subjects. There was a main effect of task at five different latencies: 148 ms (occipital), 180 ms (parietal), 388 ms (vertex), 556 ms (central-parietal), and 900 ms (vertex). Source estimations derived from topographic data indicate that frontal brain regions were strongly activated after 300 ms in the control task, but not until about 900 ms in the rotation task. We conclude that the neural computations underlying mental hand rotation may be recruited from relatively early stages of visuo-perceptual analysis; these early computations influence subsequent processing within a parietal-prefrontal system for the integration of perception with action.

Abstract 1118 There are no currently available tests to rapidly assess and reliably identify both impaired hemostasis and hypercoagulable states, in part because of difficulties in measuring integrated reactions in whole blood using a single sensitive and clinically useful platform. Methods like T2 magnetic resonance (T2MR) can provide rich information from complex samples, such as changes in the blood during hemostasis, by measuring signals emanating from the hydrogen nuclei within the sample, primarily in water. We used a 14”x6”x7” portable instrument to measure changes in T2MR that provide a continuous report on the dynamically changing microscopic environment of water during coagulation of whole blood (WB), platelet-poor or platelet-rich plasma (PRP). In these initial foundational studies, we measured T2MR continuously over 20 mins using 34 μL blood samples from consented normal adult donors. We tested clotting of WB initiated by the addition of thrombin or kaolin + calcium. Platelet activation was achieved in WB by addition of ADP or arachidonic acid in the presence of reptilase and factor XIIIa with and without addition of the platelet inhibitors aspirin or 2-methylthioadenosine 5'-monophosphate (2-MeSAMP) and results were confirmed by standard platelet aggregometry. At normal platelet counts from 1.5–3×105/μL and normal hematocrit (HCT) from 38%-48%, T2MR gave two curves corresponding to: (1) water trapped within a retracted clot and (2) water in the surrounding liquid, i.e. serum (Fig. 1a). Platelet counts Our proof-of-principle studies show that T2MR technology can be applied to measurement of blood clotting across a range of hemostatic conditions. This single technology may be applicable to the study, diagnosis, and management of a spectrum of disorders that range from impaired hemostasis to hypercoagulable states. These T2MR studies represent the first application of this technology to hemostasis and thrombosis. Additional studies will be needed to develop a more complete understanding of the biochemical events measured by T2MR and to more fully explore its clinical utility. Figure 1. Examples of T2MR 3D surfaces for 34 μL of citrated whole blood mixed activated by adding a dried formulation of CaCl2 (11 mM) and kaolin ( Disclosures: Cines: T2 Biosystems: Research Funding. Lebedeva:T2 Biosystems: Research Funding. Massefski:T2 Biosystems: Employment. Papkov:T2 Biosystems: Employment. Thayer:T2 Biosystems: Employment. Lowery:T2 Biosystems: Employment.

In order to characterize the tandem rrnB promoters transcribing one of the ribosomal RNA operons in E. coli we subcloned the basic promoter unit. This 185 bp fragment extends from -64 to +121 counted from the transcription start site of upstream promoter P1. The start site of downstream promoter P2 is also included in the promoter cartridge. S1 mapping experiments show that both promoters on this fragment are active in vivo. BAL-31 deletion mutations generated at the start site for promoter P2 were also tested by S1 mapping. Transcription from P2 remained active in all cases with the exception of one construction which lacks the -10 region. This demonstrates that the sequences downstream from the -10 region of P2 are not essential for basic promoter function.

IN REVIEWING the literature of infectious synovitis it is observed that the etiological agent or agents causing the disease remain in doubt. Olson (1956) has reported isolating a large particle virus, as well as pleuropneumonia like organisms (PPLO) and two unidentified viruses from disease outbreaks. Cover (1956) has described the etiological agent as a virus or virus-like organism, and Cover and Benton (1957) have recently been able to isolate the agent from many of the internal organs including the testes and ovary. Leece et al. (1955) cultivated the virus on heart fibroblasts. Wills and Delaplane (1955) suggested possible hatching egg transmission in one experiment. In reference to the control of the disease, differences in results have occurred with both furazolidone (nf-180) and antibiotics. Olson et al. (1956a) reported that in birds inoculated with two of seven strains of infectious synovitis, the infection could not be controlled by feeding nf-180 at . . .

NADH-cytochrome b5 oxidoreductase (Ncb5or) is an endoplasmic reticulum (ER)-associated redox enzyme involved in fatty acid metabolism, and phenotypic abnormalities of Ncb5or(-/-) mice include diabetes and lipoatrophy. These mice are lean and insulin-sensitive but become hyperglycemic at age 7 weeks as a result of β-cell dysfunction and loss. Here we examine early cellular and molecular events associated with manifestations of β-cell defects in Ncb5or(-/-) mice. We observe lower islet β-cell content in pancreata at age 4 weeks and prominent ER distention in β-cells by age 5 weeks. Ultrastructural changes progress rapidly in severity from age 5 to 6 weeks, and their frequency rises from 10% of β-cells at 5 weeks to 33% at 6 weeks. These changes correlate temporally with the onset of diabetes. ER stress responses and lipid load in Ncb5or(-/-) β-cells were assessed with isolated islets from mice at age 5 weeks. Expression levels of the stress marker protein Grp78/BiP and of phosphorylated eIF2α protein were found to be reduced, although their transcript levels did not decline. This pattern stands in contrast to the canonical unfolded protein response. Ncb5or(-/-) β-cells also accumulated higher intracellular levels of palmitate and other free fatty acids and exhibited greater reactive oxygen species production than wild-type cells. An alloxan-susceptible genetic background was found to confer accelerated onset of diabetes in Ncb5or(-/-) mice. These findings provide the first direct evidence that manifestations of diabetes in lean Ncb5or(-/-) mice involve saturated free fatty acid overload of β-cells and ER and oxidative stress responses.

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C ATCH PHRASES CAPTURE some of the flavor of "new administration." Among others, these include "open systems," "temporary," "changing," "dialectical," and "problem-solving" organizations, and "existential executives." The approaches place at least as much emphasis upon interactions between organizations and their external environments as upon the internal workings of complex organizations, because the single organization, whatever its comprehensiveness, can accomplish little by itself. Because the interactions include individuals and organizations without formal authority to order each other around, concepts of hierarchy and executive responsibility provide insufficient operational guidance. There is a need, now in the early stages of articulation, to reorient public policy processes in these directions; to establish, for example, a "variety of mechanisms that link national and local governments, academia, and the business community," among other circles of influence.1 Whatever the extent of demands from individual citizens and groups that they be able to participate, the demands seem linked more to policy than to electoral processes.2 The trends may portend, perhaps for the first time, a true synthesis of politics and administration, and the conceptualization and implementation of new processes may be the most important items on the agenda. This is because policy processes can determine who participates and, if properly designed, can decrease the likelihood of poor decisions and make it easier to reverse such decisions when they are made. For the most part, however, these new approaches have not been put to work in the most important policy processes. Recommendations for change in presidential processes, for example, tend to follow traditional models. One sees demands that the Department of State "take charge" of foreign policy, or that a "Super Secretary" standing "above" the Cabiet handle both foreign and military affairswith the assistance of a "strong" White House staff.3 Predictions of domestic "superdepartments" modeled after the Department of Defense have been followed by presidential requests to construct such departments in the hope they will encompass all the needed "authority" and "resources." 5 This persistent drive for overhead consolidation may be marching us to the rear. The notion seems to remain that if an organization can only be made big enough, all we need do is erect walls around it, treat it as a closed system, and apply hallowed concepts of command and control. Traditional models do indeed have a strong grip; even the language of administration is inherently hierarchical, making it difficult to conceptualize alternatives or describe them when they exist. This article concentrates upon the past, present, and future implications of one such alternative. The National Security Council (NSC), statutorily established in 1947 as a formal linkage of the President with his foreign policy advisers in the Executive Branch, has functioned as an interactive system, or process, for most of its lifetime, but some of its more profound impli

From one of the most popular text-books 1 on midwifery in our medical colleges, I quote the following: "The intimate connection between ovulation and menstruation is now admitted by most physiologists, and it is held that the determining cause of the discharge is the periodic maturation of the Graafian follicles. and when the ovaries are removed by operation, menstruation does not take place." One of our latest physiologies, also popular as a text-book 2 in our medical colleges, states the following: "Women do not menstruate when both ovaries have been removed by operation. Some instances have been recently recorded, indeed, of a sanguineous discharge, occurring periodically from the vagina after both ovaries have been removed for disease; and it has been inferred from this that menstruation is a function independent from the ovaries, but this evidence is not conclusive." Most of us have been educated along the line of the