Search

Apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is silenced by promoter methylation in many types of tumors, yet ASC's role in most cancers remains unknown. Here, we show that ASC is highly expressed in a model of medulloblastoma, the most common malignant pediatric brain cancer; ASC is also expressed in human medulloblastomas. Importantly, while ASC deficiency did not affect normal cerebellar development, ASC knockout mice on the Smoothened (ND2:SmoA1) transgenic model of medulloblastoma exhibited a profound reduction in medulloblastoma incidence and a delayed tumor onset. A similar decrease in tumorigenesis with ASC deficiency was also seen in the hGFAP-Cre:SmoM2 mouse model of medulloblastoma. Interestingly, hyperproliferation of the external granule layer (EGL) was comparable at P20 in both wild-type and ASC-deficient SmoA1 mice. However, while the apoptosis and differentiation markers remained unchanged at this age, proliferation makers were decreased, and the EGL was reduced in thickness and area by P60. This reduction in proliferation with ASC deficiency was also seen in isolated SmoA1 cerebellar granule precursor cells in vitro, indicating that the effect of ASC deletion on proliferation was cell autonomous. Interestingly, ASC-deficient SmoA1 cerebella exhibited disrupted expression of genes in the transforming growth factor-β pathway and increased level of nuclear Smad3. Taken together, these results demonstrate an unexpected role for ASC in Sonic hedgehog-driven medulloblastoma tumorigenesis, thus identifying ASC as a promising novel target for antitumor therapy.

Objective: We studied the complications and timing implications of prehospital activated charcoal (PAC). Appropriateness of PAC administration was also evaluated., Methods: We retrospectively reviewed prehospital records over 32 months for overdose cases, where PAC was administered. Cases were assessed for amount and type of ingestant, clinical findings, timing of PAC, timing of transport and arrival into the emergency department (ED), and complications. Encounter duration in cases of PAC was compared with that, for all cases during the study period, where an overdose patient who did not receive activated charcoal was transported., Results: Two thousand eight hundred forty-five total cases were identified. In 441 cases, PAC was given; and complications could be assessed. Two hundred eighty-one of these had complete information regarding timing of ingestion, activated charcoal administration, and transport. The average time between overdose and PAC was 49.8 minutes (range, 7-199 minutes; median, 41.0 minutes; SD, 30.4 minutes). Complications included emesis (7%), declining mental status (4%), declining blood pressure (0.4%), and declining oxygen saturation (0.4%). Four hundred seventeen cases of PAC had documentation of timing of emergency medical service (EMS) arrival on scene and arrival at the ED. Average EMS encounter time was 29 minutes (range, 10-53 minutes; median, 27.9 minutes). Two thousand forty-four poisoning patients were transported who did not receive PAC. The average EMS encounter time for this group was 28.1 minutes (range, 4-82 minutes; median, 27.3 minutes), not significantly different (P =.114)., Conclusions: Prehospital activated charcoal did not appear to markedly delay transport or arrival of overdose patients into the ED and was generally safe., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Pharmacodynamics and toxicodynamics are the study of the biochemical and physiological effects of therapeutic agents and toxicants and their mechanisms of action. MALDI-MS imaging offers great potential for the study of pharmaco/toxicodynamic responses in tissue owing is its ability to study multiple biomarkers simultaneously in a label-free manner. Here, existing examples of such studies examining anticancer drugs and topically applied treatments are described. Examination of the literature shows that the use of MS imaging in pharmaco/toxicodynamic studies is in fact quite low. The reasons for this are discussed and potential developments in the methodology that might lead to its further use are described.

Objectives: Physiologic and immunologic changes in pregnancy result in increased susceptibility to infection. These shifts are more pronounced in pregnancies complicated by multiple gestation. The objective of this study was to determine the association between multiple gestation and risk of infectious morbidity., Study Design: The Nationwide Inpatient Sample for the years 2008-2010 was used to identify pregnant women during admission for delivery with International Classification of Diseases codes. Logistic regression was used to compute odds ratios and 95% confidence intervals for demographic data, preexisting medical conditions, and acute medical and infectious complications for women with multiple versus singleton gestations., Results: Among women with multiple gestation, 38.4 per 1,000 women had an infectious complication compared to 12.8 per 1,000 women with singletons. The most significant infectious morbidity associated with multiple gestation was intestinal infections, pyelonephritis, influenza, and pneumonia. After controlling for confounding variables, infectious complications at delivery persisted for women with multiples, though the association was dependent on mode of delivery., Conclusions: Women with multiple gestations are at increased risk for infectious morbidity identified at the time of delivery. This association was diminished among women who had a cesarean suggesting that operative delivery is not responsible for this association.

HIV-1 and CMV are important pathogens transmitted via breastfeeding. Furthermore, perinatal CMV transmission may impact growth and disease progression in HIV-exposed infants. Although maternal antiretroviral therapy reduces milk HIV-1 RNA load and postnatal transmission, its impact on milk CMV load is unclear. We examined the relationship between milk CMV and HIV-1 load (4-6 weeks postpartum) and the impact of antiretroviral treatment in 69 HIV-infected, lactating Malawian women and assessed the relationship between milk CMV load and postnatal growth in HIV-exposed, breastfed infants through six months of age. Despite an association between milk HIV-1 RNA and CMV DNA load (0.39 log(10) rise CMV load per log(10) rise HIV-1 RNA load, 95% CI 0.13-0.66), milk CMV load was similar in antiretroviral-treated and untreated women. Higher milk CMV load was associated with lower length-for-age (-0.53, 95% CI: -0.96, -0.10) and weight-for-age (-0.40, 95% CI: -0.67, -0.13) Z-score at six months in exposed, uninfected infants. As the impact of maternal antiretroviral therapy on the magnitude of postnatal CMV exposure may be limited, our findings of an inverse relationship between infant growth and milk CMV load highlight the importance of defining the role of perinatal CMV exposure on growth faltering of HIV-exposed infants.

Environmental toxicants such as methylmercury play a critical role in the pathogenesis of many neurodevelopmental disorders. Environmental exposure to methylmercury frequently occurs at low doses, most frequently through fish consumption. Although the general population is at risk for exposure, pregnant women and young children are the most vulnerable. A common symptom of perinatal exposure to methylmercury is increased sensory (visual) deficits, motor impairment, and an overall cognitive decline. Research has indicated that the developing cerebellum, specifically the cerebellar granular layer, is particularly vulnerable to methylmercury neurotoxicity. This review examines the effects of low-level methylmercury exposure on motor coordination. We specifically focus on the role of cerebellar granule cells in methylmercury neurotoxicity. We suggest that methylmercury induces oxidative stress in cerebellar granule cells, which subsequently results in apoptotic cell death. Understanding the mechanism by which methylmercury induces toxicity within the developing brain will allow for enhanced treatments and potential reversal of the detrimental effects., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Background: The use of continuous positive airway pressure (CPAP) assisted ventilation in the emergency department(ED) has been well described., Objectives: The purpose of this study was to measure the efficacy of adding pre-hospital CPAP to an urban emergency medical service (EMS) respiratory distress protocol on persons with respiratory distress., Methods: A historical cohort analysis of consecutive patients between 2005 and 2010. Groups were matched for severity of respiratory distress. Physiologic variables were the primary outcome obtained from first responders and upon triage in the ED. Additional outcomes included endotracheal intubation rate, hospital mortality, overall hospital length of stay(LOS), intensive care unit (ICU) admission, and ICU length of stay (ICU LOS)., Results: There were 410 consecutive patients with predetermined criteria for severe respiratory distress, 235 historical controls matched with 175 post-implementation patients. Average age was 67 years, 54% being male. There were significant median differences in heart and respiratory rates favoring the historical cohort (p < 0.05). There were no significant differences in intubation rate, overall hospital LOS, ICU admission rate, ICU LOS, and hospital mortality (p > 0.05).Patients that were continued on noninvasive ventilatory assistance had a significantly improved rate of intubation and ICU LOS (p < 0.05)., Conclusions: The addition of CPAP to our pre-hospital respiratory distress protocol did not improve physiologic variables.There were no differences in overall and ICU LOS between groups. Persons with apparent continued ventilatory assistance appeared to have improved rates of intubation and ICU LOS [corrected]., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Worker polyphenisms in ants enable insightful analyses of neuronal underpinnings of division of labor, a crucial aspect of animal social organization. In the ant Pheidole dentata, which has a dimorphic worker caste, serotonin titer increases in the brain with age, modulating pheromonal recruitment communication and foraging, behaviors characteristic of mature individuals. Serotonin-immunoreactive (5HT-IR) neurons are found in the mushroom bodies (MB) and may modulate multi-sensory information processing associated with cues and social signals guiding task performance. The volume of this neuropil correlates with worker subcaste and age in P. dentata, but the role of structural variation in individual extrinsic MB neurons in division of labor in ants is poorly understood. We tested the hypothesis that branching complexity in a 5HT-IR calyx input neuron (CIN) in the MBs increases with age in minor workers of P. dentata in association with task repertoire expansion. We further predicted that major workers, which are defense specialists, have less elaborate CIN axonal arbors at any age in comparison to minor workers, which are task generalists. Contrary to our predictions, immunohistochemical and morphometric analyses revealed significantly greater CIN branching in both newly eclosed and mature major workers, and identified an effect of worker age on branching complexity only in majors. Our results indicate a modulatory role of the CIN in subcaste-specific behaviors and suggest behavioral specialization may be associated with the elaboration of specific MB neurons., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Quantitative neuropathologic methods provide information that is important for both research and clinical applications. The technologic advancement of digital pathology and image analysis offers new solutions to enable valid quantification of pathologic severity that is reproducible between raters regardless of experience. Using an Aperio ScanScope XT and its accompanying image analysis software, we designed algorithms for quantitation of amyloid and tau pathologies on 65 β-amyloid (6F/3D antibody) and 48 phospho-tau (PHF-1)-immunostained sections of human temporal neocortex. Quantitative digital pathologic data were compared with manual pathology counts. There were excellent correlations between manually counted and digitally analyzed neuropathologic parameters (R² = 0.56-0.72). Data were highly reproducible among 3 participants with varying degrees of expertise in neuropathology (intraclass correlation coefficient values, >0.910). Digital quantification also provided additional parameters, including average plaque area, which shows statistically significant differences when samples are stratified according to apolipoprotein E allele status (average plaque area, 380.9 μm² in apolipoprotein E [Latin Small Letter Open E]4 carriers vs 274.4 μm² for noncarriers; p < 0.001). Thus, digital pathology offers a rigorous and reproducible method for quantifying Alzheimer disease neuropathologic changes and may provide additional insights into morphologic characteristics that were previously more challenging to assess because of technical limitations.

Background: A small group of adults disproportionately and ineffectively use acute services including emergency medical services (EMS) and emergency departments (EDs). The resulting episodic, uncoordinated care is of lower quality and higher cost and simultaneously consumes valuable public safety and acute care resources., Objective: To address this issue, we measured the impact of a pilot, EMS-based case management and referral intervention termed the San Diego Resource Access Program (RAP) to reduce EMS, ED, and inpatient (IP) visits., Methods: This was a historical cohort study of RAP records and billing data of EMS and one urban hospital for 51 individuals sequentially enrolled in the program. The study sample consisted of adults with ≥ 10 EMS transports within 12 months and others reported by prehospital personnel with significant recent increases in transports. Data were collected over a 31-month time period from December 2006 to June 2009. Data were collected for equal pre- and postenrollment time periods based on date of initial RAP contact, and comparisons were made using the Wilcoxon signed-rank test. Overall use for subjects is reported., Results: The majority of subjects were male (64.7%), homeless (58.8%), and 40 to 59 years of age (72.5%). Between the pre and post periods, EMS encounters declined 37.6% from 736 to 459 (p = 0.001), resulting in a 32.1% decrease in EMS charges from $689,743 to $468,394 (p = 0.004). The EMS task time and mileage decreased by 39.8% and 47.5%, respectively, accounting for 262 (p = 0.008) hours and 1,940 (p = 0.006) miles. The number of ED encounters at the one participating hospital declined 28.1% from 199 to 143, which correlated with a 12.7% decrease in charges from $413,410 to $360,779. The number of IP admissions declined by 9.1% from 33 to 30, corresponding to a 5.9% decrease in IP charges from $687,306 to $646,881. Hospital length of stay declined 27.9%, from 122 to 88 days. Across all services, total charges declined by $314,406., Conclusions: This pilot study demonstrated that an EMS-based case management and referral program was an effective means of decreasing EMS transports by frequent users, but had only a limited impact on use of hospital services.

Crohn's disease (CD) has a bimodal distribution in incidence, with a second peak in the elderly. However, its diagnosis in the elderly is difficult due to a wider range of more common differential diagnoses such as diverticulitis, ischaemic colitis and colorectal cancer. We report a likely case of CD in a 95-year-old woman. She presented with diarrhoea and rectal bleeding and was found to have multiple pleomorphic ulcers with a patchy cobblestone mucosa on sigmoidoscopy. Histopathology demonstrated focal ulceration, altered crypt architecture and adjacent neutrophil polymorph infiltration with no granolomata or features of malignancy. The patient passed away after steroid treatment was started. This case is a reminder that CD can present in the elderly and highlights the challenging diagnosis and high mortality of CD-related hospitalisation in the elderly. When considering management, attention should be given to comorbid disease, age-related changes in pharmacokinetics and patients social circumstances.

Objectives: This study sought to measure the impact of pre-hospital (PH) electrocardiography (ECG) on scene-to-hospital time for patients with chest pain of cardiac origin and those with ST-segment elevation myocardial infarction (STEMI)., Background: Pre-hospital ECG decreases door-to balloon (D2B) time for STEMI patients. However, obtaining a PH ECG might prolong scene time. We investigated the impact of obtaining a PH ECG on both scene and transport times for patients with chest pain suspected of cardiac origin., Methods: City of San Diego Emergency Medical System runsheets of patients with chest pain from January 2003 to April 2008 were analyzed. The scene times and transport times were compared before (from January 2003 to December 2005) and after (from January 2006 to April 2008) implementation of the PH ECG. Among patients with a PH ECG, median scene times and transport times were compared in patients with and without STEMI., Results: There were 21,742 patients evaluated for chest pain during the study period. Implementation of PH ECG resulted in minimal increases in median scene time (19 min, 10 s vs. 19 min, 28 s, p = 0.002) and transport time (13 min, 16 s vs. 13 min, 28 s, p = 0.007). However, compared with chest pain patients, in STEMI patients (n = 303), shorter median scene time (17 min, 51 s vs. 19 min, 31 s, p < 0.001), transport time (12 min, 34 s vs. 13 min, 31 s, p = 0.006), and scene-to-hospital time was observed (30 min, 45 s vs. 33 min, 29 s, p < 0.001)., Conclusions: Obtaining a PH ECG for patients with chest pain minimally prolongs scene and transport times. Further, for STEMI patients, both scene times and transport times are actually reduced leading to a potential reduction in total ischemic time., (Copyright © 2012 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.)

Background: Recent studies have described a gender bias against women in the setting of acute coronary syndrome (ACS)., Objectives: We sought to measure the impact that a prehospital electrocardiogram (PH ECG) has on prehospital total scene time to hospital arrival time, comparing men and women with the complaint of chest pain (cCP)., Methods: This study retrospectively analyzed San Diego Emergency Medical Services (EMS) runsheets of patients with cCP before and after implementation of the PH ECG protocol. The average scene time (ST), transport time (TT), and total scene-to-arrival-at-hospital time (STH) were compared. After stratification by gender, times were compared in patients with ST-elevation myocardial infarction (STEMI) to those without STEMI., Results: Of 21,742 EMS activations for patients with cCP, there were no significant differences overall. When stratified by gender, there was a significant reduction of ST (00:19:16 min vs. 00:20:48 min, p<0.001, 95% CI 00:01:17-00:01:48) and STH (00:33:22 min vs. 00:35:44 min, p<0.001, 95% CI 00:01:21-00:02:24) favoring men in cases without STEMI. In cases of STEMI, men had a significant reduction in ST (00:17:27 min vs. 00:20:29 min, p<0.001, 95% CI 00:01:24-00:04:40) and STH (00:30:30 min vs. 00:34:25 min, p<0.01, 95% CI 00:01:23-00:06:26) times compared to women., Conclusion: Prehospital ECG implementation led to no significant differences in pre- and post-implementation times. In cases of STEMI, men had significantly reduced scene time and scene-to-hospital time when compared to women. The precise reason for these disparities remains unknown., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The Institute for Healthcare Improvement Open School for Health Professions is an international organization that provides the next generation of health-care leaders with the skills to lead improvement in health care. This article discusses how doctors can get involved and implement change at their hospital.

Background: Given the frequency with which many different strains of Staphylococcus aureus are found in various prehospital settings, this study sought to characterize S aureus isolates taken from one such environment. The objectives were to determine the frequency of S aureus in front-line, advanced life support (ALS) ambulances throughout the Chicago metropolitan area, and to generate antibiograms (antibiotic resistance profiles) for each S aureus isolate using 8 clinically relevant antibiotics., Methods: Samples were obtained from 26 sites in 71 ambulances from 34 different Chicago-area municipalities. Selected colonies that demonstrated a growth pattern consistent with that of S aureus were subjected to a latex agglutination test specific for S aureus. Antibiograms and genetic analyses were performed on all latex agglutination test-positive isolates., Results: At least one S aureus isolate was found in approximately 69% of all ambulances in the study. Of all isolates detected, 77% showed resistance to at least one antibiotic, and 34% displayed resistance to 2 or more antibiotics. Some level of oxacillin resistance was found in 21% of isolates; however, only slightly more than half of these oxacillin-resistant isolates were found to carry the methicillin-resistant S aureus-specific SCCmec cassette. Some 12% of all isolates were ultimately determined to be methicillin-resistant S aureus, whereas the remaining 88% were methicillin-sensitive S aureus with varying antibiograms., Conclusions: Antibiotic resistance appears to be prevalent in S aureus isolates detected in Chicago area ALS ambulances. Given the ease with which S aureus can survive on inanimate surfaces and exchange antibiotic resistance elements, a conscientious approach to the application of existing cleaning techniques, especially in key ambulance sites, is needed. Future work will include further characterizing isolates using multiple techniques, as well as follow-up studies with interested municipalities., (Copyright © 2012 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.)

Nickel and cobalt are heavy metals found in land, water, and air that can enter the body primarily through the respiratory tract and accumulate to toxic levels. Nickel compounds are known to be carcinogenic to humans and animals, while cobalt compounds produce tumors in animals and are probably carcinogenic to humans. People working in industrial and manufacturing settings have an increased risk of exposure to these metals. The cytotoxicity of nickel and cobalt has individually been demonstrated; however, the underlying mechanisms of co-exposure to these heavy metals have not been explored. In this study, we investigated the effect of exposure of H460 human lung epithelial cells to nickel and cobalt, both alone and in combination, on cell survival, apoptotic mechanisms, and the generation of reactive oxygen species and double strand breaks. For simultaneous exposure, cells were exposed to a constant dose of 150 μM cobalt or nickel, which was found to be relatively nontoxic in single exposure experiments. We demonstrated that cells exposed simultaneously to cobalt and nickel exhibit a dose-dependent decrease in survival compared to the cells exposed to a single metal. The decrease in survival was the result of enhanced caspase 3 and 7 activation and cleavage of poly (ADP-ribose) polymerase. Co-exposure increased the production of ROS and the formation of double strand breaks. Pretreatment with N-acetyl cysteine alleviated the toxic responses. Collectively, this study demonstrates that co-exposure to cobalt and nickel is significantly more toxic than single exposure and that toxicity is related to the formation of ROS and DSB., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Previous studies examining methods of monitoring the training and match load in soccer players have simply compared those methods to each other, not to changes in fitness. Training and match load measures from nine professional youth soccer players were collected for a period of six weeks. A lactate threshold test was conducted before and after this period. Mean weekly training and match load as determined by session-RPE, Banister's TRIMP, Team TRIMP and individualised TRIMP (iTRIMP) were correlated with each other, percentage changes in the velocity at 2 mmol · L(-1) (vLT) and 4 mmol · L(-1) (vOBLA) blood lactate concentration, and heart rate at 2 mmol · L(-1) (LT(HR)) and 4 mmol · L(-1) (OBLA(HR)). There were no significant changes in fitness across the six weeks: vLT (p = 0.54), vOBLA (p = 0.16), LT(HR) (p = 0.51) and OBLA(HR) (p = 0.63). Banister's TRIMP was significantly correlated with session-RPE (r = 0.75; p = 0.02) and Team TRIMP (r = 0.92; p < 0.001). The percentage change in vLT was significantly correlated to mean weekly iTRIMP (r = 0.67; p = 0.04). The results suggest that an individualised measure of internal load (iTRIMP) related better than other methods to changes in vLT in professional youth soccer players.

Hippocampal sclerosis is a relatively common neuropathological finding (∼10% of individuals over the age of 85 years) characterized by cell loss and gliosis in the hippocampus that is not explained by Alzheimer's disease. Hippocampal sclerosis pathology can be associated with different underlying causes, and we refer to hippocampal sclerosis in the aged brain as hippocampal sclerosis associated with ageing. Much remains unknown about hippocampal sclerosis associated with ageing. We combined three different large autopsy cohorts: University of Kentucky Alzheimer's Disease Centre, the Nun Study and the Georgia Centenarian Study to obtain a pool of 1110 patients, all of whom were evaluated neuropathologically at the University of Kentucky. We focused on the subset of cases with neuropathology-confirmed hippocampal sclerosis (n=106). For individuals aged≥95 years at death (n=179 in our sample), each year of life beyond the age of 95 years correlated with increased prevalence of hippocampal sclerosis pathology and decreased prevalence of 'definite' Alzheimer's disease pathology. Aberrant TAR DNA protein 43 immunohistochemistry was seen in 89.9% of hippocampal sclerosis positive patients compared with 9.7% of hippocampal sclerosis negative patients. TAR DNA protein 43 immunohistochemistry can be used to demonstrate that the disease is usually bilateral even when hippocampal sclerosis pathology is not obvious by haematoxylin and eosin stains. TAR DNA protein 43 immunohistochemistry was negative on brain sections from younger individuals (n=10) after hippocampectomy due to seizures, who had pathologically confirmed hippocampal sclerosis. There was no association between cases with hippocampal sclerosis associated with ageing and apolipoprotein E genotype. Age of death and clinical features of hippocampal sclerosis associated with ageing (with or without aberrant TAR DNA protein 43) were distinct from previously published cases of frontotemporal lobar degeneration TAR DNA protein 43. To help sharpen our ability to discriminate patients with hippocampal sclerosis associated with ageing clinically, the longitudinal cognitive profile of 43 patients with hippocampal sclerosis associated with ageing was compared with the profiles of 75 controls matched for age, gender, education level and apolipoprotein E genotype. These individuals were followed from intake assessment, with 8.2 (average) longitudinal cognitive assessments. A neuropsychological profile with relatively high-verbal fluency but low word list recall distinguished the hippocampal sclerosis associated with ageing group at intake (P<0.015) and also 5.5-6.5 years before death (P<0.005). This may provide a first step in clinical differentiation of hippocampal sclerosis associated with ageing versus pure Alzheimer's disease in their earliest stages. In summary, in the largest series of autopsy-verified patients with hippocampal sclerosis to date, we characterized the clinical and pathological features associated with hippocampal sclerosis associated with ageing.

Dendritic cells (DCs) play a key role in innate and adaptive immunity but the access to sufficient amount of DCs for basic and translational research has been limited.We established a novel ex vivo system to develop and expand DCs from hematopoietic stem/progenitor cells (HPCs). Both human and mouse HPCs were expanded first in feeder culture supplemented with c-Kit ligand (KL, stem cell factor, steel factor or CD117 ligand), Flt3 ligand (fms-like tyrosine kinase 3, Flt3L, FL), thrombopoietin (TPO), IL-3, IL-6, and basic fibroblast growth factor (bFGF), and then in a second feeder culture ectopically expressing all above growth factors plus GM-CSF and IL-15.In the dual culture system, CD34+ HPCs differentiated toward DC progenitors (DCPs), which expanded more than five orders of magnitude. The DCPs showed myeloid DC surface phenotype with up-regulation of transcription factors PU.1 and Id2, and DC-related factors homeostatic chemokine ligand 17 (CCL17) and beta-chemokine receptor 6 (CCR6). Multiplex ELISA array and cDNA microarray analyses revealed that the DCPs shared some features of IL-4 and IL-15 DCs but displayed a pronounced proinflammatory phenotype. DCP-derived DCs showed antigen-uptake and immune activation functions analogous to that of the peripheral blood-derived DCs. Furthermore, bone marrow HPC-derived DCP vaccines of tumor-bearing mice suppressed tumor growth in vivo.This novel approach of generating DCP-DCs, which are different from known IL-4 and IL-15 DCs, overcomes both quantitative and qualitative limitations in obtaining functional autologous DCs from a small number of HPCs with great translational potential.

Background: It is widely reported that hexose sugars slow gastric emptying (GE) via osmoreceptor stimulation but this remains uncertain. We evaluated the effects of a panel of hexoses of differing molecular structure, assessing the effects of osmolality, intra-individual reproducibility and the role of the CCK(1) receptor, in the regulation of GE by hexoses., Methods: Thirty one healthy non-obese male and female subjects were studied in a series of protocols, using a (13) C-acetate breath test to evaluate GE of varying concentrations of glucose, galactose, fructose and tagatose, with water, NaCl and lactulose as controls. GE was further evaluated following the administration of a CCK(1) receptor antagonist. Three subjects underwent repeated studies to evaluate intra-individual reproducibility., Key Results: At 250 mOsmol, a hexose-specific effect was apparent: tagatose slowed GE more potently than water, glucose and fructose (P < 0.05). Fructose (P < 0.05) also slowed GE, but with substantial inter-, but not intra-, individual differences. As osmolality increased further the hexose-specific differences were lost. At 500 mOsmol, all hexoses slowed GE compared with water (P < 0.05), whereas lactulose and saline did not. The slowing of GE by hexose sugars appeared to be CCK(1) receptor-dependent., Conclusions & Inferences: The effects of hexose sugars on GE appear related to their molecular structure rather than osmolality per se, and are, at least in part, CCK(1) receptor-dependent., (© 2010 Blackwell Publishing Ltd.)

We evaluated the association between mini-mental status examination (MMSE) scores proximal to death and the values of 43 different clinical and pathological parameters. Studies were performed using data from 334 elderly, longitudinally evaluated research subjects who had undergone autopsy and satisfied inclusion criteria from an initial study group of 501. Interindividual variance in MMSE scores was used as a surrogate for the severity of cognitive impairment linked to aging (CILA). A statistical linear regression-based model provided a framework for assessing the parameters with significant, direct impact on CILA severity. Strong association between CILA and Alzheimer's disease (AD) pathology, especially isocortical neurofibrillary tangles, was evident. The pattern of association between AD lesion densities with cognitive impairment severity was biologically informative, with neuritic plaques having more impact in relatively high-functioning individuals. Abundant isocortical Lewy bodies tended to be an additive pathology correlating with final MMSE scores approximately 10 points lower. In a subset of cases we found evidence for association between TDP-43-related pathology and CILA severity, independent of AD or hippocampal sclerosis. There was no support for independent association between CILA severity and most evaluated indices including diffuse plaques, argyrophilic grains, heart disease, education level, apolipoprotein E alleles or diabetes.

Human haematopoietic progenitor/stem cells (HPCs) differentiate into functional T cells in the thymus through a series of checkpoints. A convenient in vitro system will greatly facilitate the understanding of T-cell development and future engineering of therapeutic T cells. In this report, we established a lentiviral vector-engineered stromal cell line (LSC) expressing the key lymphopoiesis regulator Notch ligand, Delta-like 1 (DL1), as feeder cells (LSC-mDL1) supplemented with Flt3 ligand (fms-like tyrosine kinase 3, Flt3L or FL) and interleukin-7 for the development of T cells from CD34(+) HPCs. We demonstrated T-cell development from human HPCs with various origins including fetal thymus (FT), fetal liver (FL), cord blood (CB) and adult bone marrow (BM). The CD34(+) HPCs from FT, FL and adult BM expanded more than 100-fold before reaching the beta-selection and CD4/CD8 double-positive T-cell stage. The CB HPCs, on the other hand, expanded more than 1000-fold before beta-selection. Furthermore, the time required to reach beta-selection differed for the various HPCs, 7 days for FT, 14 days for FL and CB, and 35 days for adult BM. Nevertheless, all of the T cells developed in vitro were stalled at the double-positive or immature single-positive stage with the exception that some CB-derived T cells arrived at a positive selection stage. Consequently, the LSC-mDL1 culture system illustrated diverse T-cell development potentials of pre- and post-natal and adult human BM HPCs. However, further modification of this in vitro T-cell development system is necessary to attain fully functional T cells.

Background: Recognizing factors that cause prehospital stroke delays may improve time of presentation to the Emergency Department (ED) and allow earlier treatment of acute stroke patients., Study Objectives: To determine the impact of stroke recognition by emergency medical dispatchers (EMD) and paramedics (PM) on ED arrival time in a large urban Emergency Medical Services system., Methods: Retrospective study of patients aged 18 years or more identified as having acute stroke by EMD, PM, or stroke neurologists from January 1, 2005 to December 31, 2005. Data were acquired from computer-assisted dispatch records, paramedic assessments, ICD-9 (International Classification of Diseases, 9(th) Revision) databases, and a hospital stroke registry. Paramedic time to scene, scene time, and total run time were computed for patients with final hospital diagnosis of stroke and grouped into missed strokes and identified strokes by EMD and PM. Time intervals were compared between missed and identified strokes as well as between incidents where EMD and PM agreed or disagreed., Results: A total of 1067 patients were eligible for the study; 22 were excluded for missing data. For true strokes, EMD and PM were in agreement 27.3% of the time. The median RT was 2.5 min shorter when there was agreement between the providers than when there was disagreement (36.5 min; interquartile range [IQR] 30-43 vs. 39 min.; IQR 33-45, respectively)., Conclusions: Prehospital scene time and run times for acute strokes are less when there is diagnostic concordance between dispatchers and paramedics. Time intervals did not differ between missed and recognized strokes.

Brains that have many neurofibrillary tangles (NFTs) in medial temporal lobe structures (Braak stage III or IV) but no cortical neuritic plaques (NPs) may be a diagnostic dilemma; they also raise questions about the amyloid cascade hypothesis of Alzheimer disease (AD) in which NFT development is thought to occur downstream of the development of amyloid plaques. To determine the clinical, demographic, and biological factors related to NFT+/NP- cases, we analyzed 26 NFT+/NP- patient brains identified from the University of Kentucky AD Center autopsy cohort (n=502); most of these patients were at least 85 years old and lacked profound antemortem cognitive impairment. A subset of the cases had NFTs in the medulla oblongata. Aberrant trans-activator regulatory DNA-binding protein 43 immunohistochemical staining was seen in 5 of the 26 cases with the clinical diagnoses of AD or mild cognitive impairment. We also queried cases in the National Alzheimer's Coordinating Center Registry (n=5,108) and found 219 NFT+/NP- cases. Those patients had a relatively high likelihood of belonging to a birth cohort with the highest incidence of influenza infection during the 1918 to 1919 pandemic. This observation may link the pathogenesis in NFT+/NP- cases to encephalitis during childhood. We conclude that NFT+/NP- cases comprise approximately 5% of aged individuals in multiple data sets; these cases are not necessarily within the spectrum of AD.

Natural Abs, which arise without known immune exposure, have been described that specifically recognize cells dying from apoptosis, but their role in innate immunity remains poorly understood. Herein, we show that the immune response to neoantigenic determinants on apoptotic thymocytes is dominated by Abs to oxidation-associated Ags, phosphorylcholine (PC), a head group that becomes exposed during programmed cell death, and malondialdehyde (MDA), a reactive aldehyde degradation product of polyunsaturated lipids produced following exposure to reactive oxidation species. While natural Abs to apoptotic cells in naive adult mice were dominated by PC and MDA specificities, the amounts of these Abs were substantially boosted by treatment of mice with apoptotic cells. Moreover, the relative amounts of PC and MDA Abs was affected by V(H) gene inheritance. Ab interactions with apoptotic cells also mediated the recruitment of C1q, which enhanced apoptotic cell phagocytosis by immature dendritic cells. Significantly, IgM Abs to both PC and MDA were primary factors in determining the efficiency of serum-dependent apoptotic cell phagocytosis. Hence, we demonstrate a mechanism by which certain natural Abs that recognize neoantigens on apoptotic cells, in naive mice and those induced by immune exposure to apoptotic cells, can enhance the functional capabilities of immature dendritic cells for phagocytic engulfment of apoptotic cells.

We studied Alzheimer's disease (AD) pathology in the precuneus and surrounding brain areas. Anatomically, the precuneus corresponds to the medial portion of human cerebral cortical Brodmann Area 7. This study utilized patients from the University of Kentucky Alzheimer's Disease Center autopsy cohort. Data from 47 brains were used comprising patients of differing antemortem cognitive impairment severities, each with longitudinal clinical data and extensive neuropathological data. We assessed whether the precuneus and surrounding areas are differentially vulnerable to AD-type pathological lesions (diffuse amyloid plaques, neuritic amyloid plaques, and neurofibrillary tangles). Eleven areas of brain were evaluated for each case: amygdala, hippocampal CA1, subiculum, entorhinal cortex, frontal cortex, superior and middle temporal gyri, inferior parietal lobule, occipital cortex, posterior cingulate gyrus, Brodmann Area 31, and the precuneus proper. Like other areas of neocortex, the precuneus demonstrated increased diffuse and neuritic amyloid plaques early in the evolution in AD, and increased neurofibrillary tangles late in AD. Correcting for the antemortem cognitive status of the patients, there was no evidence of an increase in the density of AD-type pathology in the precuneus or neighboring areas relative to other areas of cerebral neocortex. Our results are not consistent with the idea that the precuneus is involved in a special way with plaques or tangles relative to other areas of neocortex.

Objective: To determine the efficacy of electrocardiography (ECG) in detecting left ventricular hypertrophy (LVH) in pediatric hypertension (HT)., Study Design: Concomitant echocardiograms and electrocardiograms in 108 children with HT were reviewed. Left ventricular mass (LVM), assessed by echocardiography, was used as a basis for a diagnosis of LVH (echo LVH) using accepted pediatric criteria. Using Wilcoxon's rank-sum test, 14 ECG variables were compared between subjects with and without echo LVH. Spearman correlations were used to examine the linear association between echo LVH and these ECG variables. The sensitivity and specificity of ECG in diagnosing LVH were computed., Results: Of the 108 subjects studied, 35 (32%) met the pediatric criteria for LVH; of these, 8 (7.4%) also met the adult criteria (>51 g/m(2.7)) for LVH. Mean values for only 5 ECG criteria differed significantly among the groups: RI, SaVR, RaVL, RI+SIII, and SVI+RV6 (P < .05). Significant correlations were found for several ECG criteria and at least 1 measure of LVM, but the magnitudes were modest. Standard ECG criteria predicted LVH with high specificity (>90%) but low sensitivity (<35%). RI >10 mm was identified as demonstrating a modestly improved positive likelihood ratio of approximately 3., Conclusions: ECG is not an adequate predictor of LVH for clinical use in HT.

Background: Prehospital personnel in Emergency Medical Service (EMS) systems have varying levels of accuracy in stroke recognition. Identifying the accuracy of emergency medical dispatcher using Medical Priority Dispatch Systems (MPDS) stroke protocol and paramedics may help understand the accuracy of stroke recognition in about 3000 emergency medical dispatch systems and prehospital systems world wide., Objective: Our aim was to assess the accuracy of stroke identification in emergency medical dispatchers (EMD) with high compliance to MPDS protocol and paramedics using Cincinnati Prehospital Stroke Scale (CSS)., Methods: This was a retrospective observational study. Data was acquired from a computer assisted dispatch (CAD) system, a computerized paramedic record database and discharge diagnosis from billing records or stroke registry containing all stroke assessments of patients who presented to the participating study hospitals within 12 hours of symptom onset. We included patients 18 years or older, identified as having stroke by EMD and city agency paramedics. We excluded patients taken to hospitals not participating in the study, patients with a dispatch determinant of Stroke (card 28) not transported by City EMS agency (SDMSE) to participating hospitals, patients in the stroke registry not transported by SDMSE or patients with no final outcome data. A stroke neurologist or hospital discharge diagnosis of stroke (physician diagnosis) was used to determine the sensitivity and predictive values of EMD and paramedic recognition of stroke., Results: Of 882 patients with a dispatch determinant of stroke using MPDS Stroke protocol, 367 had a final discharge diagnosis of stroke. This gives a sensitivity of 83% and a positive predictive value of 42% for EMD using MPDS Stroke protocol. Of 477 patients with a paramedic assessment of stroke using CSS, 193 had a final discharge diagnosis of stroke. This gives a sensitivity of 44% and a PPV of 40% for paramedics using CSS., Conclusions: In our EMS system, EMD using MPDS Stroke protocol with a high compliance has a higher sensitivity than paramedics using CSS.

Amnestic mild cognitive impairment (MCI) appears to be a very early stage of Alzheimer's disease (AD). The amyloid-beta peptide (Abeta) is believed to be a possible substrate for AD, but little is currently known about Abeta alterations in MCI and how these changes compare to later stages of disease. In the present study Abeta was differentially extracted from the brains of age-matched control, MCI, and AD cases and compared with plaque counts. For comparison, APPxPS-1 knock-in mice were processed in parallel. We observed that Abeta42 was significantly elevated in MCI subjects, even though there was no significant alteration in the total amount of Abeta. Relative Abeta solubility within the different extractable pools was identical between AD and MCI subjects, with both significantly altered relative to controls. Temporal analysis of Abeta levels and solubility in a knock-in mouse model of Abeta pathogenesis recapitulated many of the salient features observed in AD. Characterization of the SDS fraction showed some similarities between aged knock-in mice and AD subjects. These data suggest that distinct changes in Abeta occur throughout the progression of AD, and that elevations in Abeta42 occur at an early, clinically defined stage.

The p38 mitogen-activated protein kinase is a stress-activated enzyme responsible for transducing inflammatory signals and initiating apoptosis. In the Alzheimer's disease (AD) brain, increased levels of phosphorylated (active) p38 were detected relative to age-matched normal brain. Intense phospho-p38 immunoreactivity was associated with neuritic plaques, neuropil threads, and neurofibrillary tangle-bearing neurons. The antibody against phosphorylated p38 recognized many of the same structures as an antibody against aberrantly phosphorylated, paired helical filament (PHF) tau, although PHF-positive tau did not cross-react with the phospho-p38 antibody. These findings suggest a neuroinflammatory mechanism in the AD brain, in which aberrant protein phosphorylation affects signal transduction elements, including the p38 kinase cascade, as well as cytoskeletal components.

Oven methods for determining moisture (volatiles) in forages and other animal feeds are empirical. The moisture concentration obtained depends upon the time and temperature the sample was dried and is influenced by the presence of other volatiles than water. A validated reference method to measure water in forages and animal feeds could be used to evaluate the appropriateness of oven methods for various types of animal feeds and forages. Karl Fischer titration is a well-established method for determining water. However, thorough extraction of water from forages and feeds is a challenge because they often contain cellular structures that release water slowly. Water was successfully extracted into methanol-formamide (50 + 50) by high-speed homogenization and then titrated directly at 50 degrees C with a one-component Karl Fischer reagent based on imidazole. The method is described in detail, results of day-to-day repeatability and laboratory-to-laboratory reproducibility are reported, and preliminary comparison data between oven methods are provided.

The goals of this study were twofold: to determine whether species differences in Abeta N-terminal heterogeneity explain the absence of neuritic plaques in the aged dog and aged bear in contrast to the human; and to compare Abeta N-terminal isoforms in parenchymal vs cerebrovascular Abeta (CVA) deposits in each of the species, and in individuals with Alzheimer disease (AD) vs nondemented individuals. N-terminal heterogeneity can affect the aggregation, toxicity, and stability of Abeta. The human, polar bear, and dog brain share an identical Abeta amino acid sequence. Tissues were immunostained using affinity-purified polyclonal antibodies specific for the L-aspartate residue of Abeta at position one (AbetaN1[D]), D-aspartate at N1 (AbetaN1[rD]), and pyroglutamate at N3 (AbetaN3[pE]) and p3, a peptide beginning with leucine at N17 (AbetaN17[L]). The results demonstrate that each Abeta N-terminal isoform can be present in diffuse plaques and CVA deposits in AD brain, nondemented human, and the examined aged animal models. Though each Abeta N-terminal isoform was present in diffuse plaques, the average amyloid burden of each isoform was highest in AD vs polar bear and dog (beagle) brain. Moreover, the ratio of AbetaN3(pE) (an isoform that is resistant to degradation by most aminopeptidases) vs AbetaN17(L)-x (the potentially nonamyloidogenic p3 fragment) was greatest in the human brain when compared with aged dog or polar bear. Neuritic plaques in AD brain typically immunostained with antibodies against AbetaN1(D) and AbetaN3(pE), but not AbetaN17(L) or AbetaN1(rD). Neuritic deposits in nondemented individuals with atherosclerotic and vascular hypertensive changes could be identified with AbetaN1(D), AbetaN3(pE), and AbetaN1(rD). The presence of AbetaN1(rD) in neuritic plaques in nondemented individuals with atherosclerosis or hypertension, but not in AD, suggests a different evolution of the plaques in the two conditions. AbetaN1(rD) was usually absent in human CVA, except in AD cases with atherosclerotic and vascular hypertensive changes. Together, the results demonstrate that diffuse plaques, neuritic plaques, and CVA deposits are each associated with distinct profiles of Abeta N-terminal isoforms.

The functional role of interleukin (IL)-4 in the development of airway hyperresponsiveness (AHR) and pulmonary eosinophilia in response to sensitization and challenge of mice with sheep red blood cells (SRBC) was examined. Control- and SRBC-sensitized A/J mice were treated with an antibody to the murine IL-4 receptor (anti-IL-4R) 3 days before intratracheal challenge with the antigen or vehicle only. Blockade of IL-4R significantly reduced antigen-induced AHR and prevented increases in goblet cells and bronchoalveolar lavage (BAL) eosinophils. Treatment with anti-IL-4R did not affect antigen-induced increases in lung mRNA and BAL protein levels of IL-5 and interferon-gamma or IL-4 mRNA but did significantly increase IL-4 protein levels. Antigen-induced AHR was not reduced by treatment with antibodies to the adhesion molecules, vascular cell adhesion molecule-1 and very late activation antigen-4. Administration of IL-4 over a 7-day period did not increase airway reactivity or induce any changes in BAL cell numbers in naive mice. These results demonstrate that IL-4 is necessary for in vivo development of antigen-induced AHR, goblet cell metaplasia, and pulmonary eosinophilia.

Immunocytochemistry, using antibodies specific for different carboxy termini of beta-amyloid. A beta 40 and A beta 42(43), was used to compare beta-amyloid deposits in aged animal models to nondemented and demented Alzheimer's disease human cases. Aged beagle dogs exhibit diffuse plaques in the absence of neurofibrillary pathology and the aged polar bear brains contain diffuse plaques and PHF-1-positive neurofibrillary tangles. The brains of nondemented human subjects displayed abundant diffuse plaques, whereas the AD cases had both diffuse and mature (cored) neuritic plaques. Diffuse plaques were positively immunostained with an antibody against A beta 42(43) in all examined species, whereas A beta 40 immunopositive mature plaques were observed only in the human brain. Anti-A beta 40 strongly immunolabeled cerebrovascular beta-amyloid deposits in each of the species examined, although some deposits in the polar bear brain were preferentially labeled with anti-A beta 42(43). beta-amyloid deposition was evident in the outer molecular layer of the dentate gyrus in the aged dog, polar bear, and human. Within this layer, A beta 42 was present as diffuse deposits, although these deposits were morphologically distinct in each of the examined animal models. In dogs, A beta 42 was cloud-like in nature; the polar bear demonstrated a more aggregated type of deposition, and the nondemented human displayed well-defined deposits. Alzheimer's disease cases were most frequently marked by neuritic plaques in this region. Taken together, the data indicate that beta-amyloid deposition in aged mammals is similar to the earliest stages observed in human brain. In each species, A beta 42(43) is the initially deposited isoform in diffuse plaques.

Aged canines exhibit central neuropathological changes strikingly similar to those seen in patients with Alzheimer's disease. In this study, brain tissue from pure bred beagles raised in a controlled environment were examined for Alzheimer-like pathology. The mean age of the animals was 15.6 years. The incidence of plaques among these 29 dogs was 65.5%. Of the 19 samples that demonstrated Alzheimer-like pathology, 18 were characterized as diffuse and one as neuritic. Plaque density was found to be independent of age. Plaque numbers were highest in the perirhinal cortex and the adjacent temporal cortex. Familial influence on plaque development is supported by congruence within 15 of the 16 litters examined (p < 0.001). In this environmentally controlled group the diffuse plaques were rarely converted to the dense neuritic plaques found in Alzheimer's disease.

Fast atom bombardment mass spectrometry and gas chromatography-mass spectrometry were used to analyze bile acids in the body fluids of an infant (L.C.) whose liver contained no immunoreactive peroxisomal 3-oxoacyl-CoA thiolase. The profiles were compared with those of six patients with undetectable peroxisomes (Zellweger syndrome) and two siblings (N.B. and I.B.) whose defect of peroxisomal beta-oxidation could not be localized by morphological studies of peroxisomes or by immunoblotting of peroxisomal beta-oxidation proteins. 3 alpha, 7 alpha, 12 alpha-Trihydroxy-5 beta-cholestan-26-oic acid (THCA) was present in bile and plasma of all patients. However, bile from L.C., N.B. and I.B. contained unconjugated varanic acid (3 alpha, 7 alpha, 12 alpha, 24-tetrahydroxy-5 beta-cholestan-26-oic acid) as the major C27 bile acid, whereas bile from Zellweger patients contained only small amounts of varanic acid. In the bile from L.C. two isomers of varanic acid were present; in the bile from N.B. and I.B. a single isomer predominated. L.C., N.B., and I.B. all produced bile containing small amounts of (24E)-3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholest-24-en-26-oic acid [( 24E]-delta 24-THCA), its [24Z]- isomer, 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholest-23-en-26-oic acid and 3 alpha, 7 alpha, 12 alpha-trihydroxy-27-nor-5 beta-cholestan-24-one. The results provide evidence for peroxisomal pathways for cholic acid synthesis in man via THCA, delta 24-THCA and varanic acid and show that bile acid analyses can be used to diagnose peroxisomal thiolase deficiency.

The properties, tissue and cellular distribution of intestinal transglutaminase have been investigated. Transglutaminase was assayed with dimethylcasein and [14C]putrescine as substrates. The enzyme has maximum activity at pH 10, although more reliable assays are made at pH 9. Transglutaminase showed an absolute requirement for Ca2+ and exhibited linear assay kinetics. The Km for putrescine was approx. 0.15 mmol/l. Tissue distribution studies suggest transglutaminase is more active in the more muscular segments of the gut. The cellular localization in jejunum was investigated by sequential cell release techniques. Approximately 2 per cent of the total activity was found in the enterocytes and crypt cells. Most of the activity was in the submucosa and serosa suggesting an interstitial cell localization. Acute hypoplastic enteropathy induced by methotrexate was accompanied by a striking decrease in mucosal transglutaminase but the activity returned to control values by 72 h. There was no significant increase in activity during the period of intense crypt cell hyperplasia and it is concluded that intestinal transglutaminase is not implicated in crypt cell proliferation.