Bioactivity-guided identification and cell signaling technology to delineate the immunomodulatory effects of Panax ginseng on human promonocytic U937 cells

<p>Abstract</p> <p>Background</p> <p>Ginseng is believed to have beneficial effects against human diseases, and its active components, ginsenosides, may play critical roles in its diverse physiological actions. However, the mechanisms underlying ginseng's effects remain to be investigated. We hypothesize some biological effects of ginseng are due to its anti-inflammatory effects.</p> <p>Methods</p> <p>Human promonocytic U937 cells were used to investigate the immunomodulatory effects of ginseng following TNF-α treatment. A global gene expression profile was obtained by using genechip analysis, and specific cytokine expression was measured by quantitative RT-PCR and ELISA. HPLC was used to define the composition of ginsenosides in 70% ethanol-water extracts of ginseng. Activation of signalling kinases was examined by Western blot analysis.</p> <p>Results</p> <p>Seventy percent ethanol-water extracts of ginseng significantly inhibited the transcription and secretion of CXCL-10 following TNF-α stimulation. Nine ginsenosides including Rb₁, Rb₂, Rc, Rd, Re, Rf, Rg₁, Rg₃and Rh₁were identified in our extract by HPLC. Seven out of nine ginsenosides could significantly inhibit TNF-α-induced CXCL-10 expression in U937 cells and give comparable inhibition of CXCL-10 transcription to those with the extract. However, the CXCL-10 suppressive effect of individual ginsenosides was less than that of the crude extract or the mixture of ginsenosides. The CXCL-10 suppression can be correlated with the inactivation of ERK1/2 pathways by ginseng.</p> <p>Conclusion</p> <p>We showed ginseng suppressed part of the TNF-α-inducible cytokines and signalling proteins in promonocytic cells, suggesting that it exerts its anti-inflammatory property targeting at different levels of TNF-α activity. The anti-inflammatory role of ginseng may be due to the combined effects of ginsenosides, contributing in part to the diverse actions of ginseng in humans.</p

Differential effects of Radix Paeoniae Rubra (Chishao) on cytokine and chemokine expression inducible by mycobacteria

<p>Abstract</p> <p>Background</p> <p>Upon initial infection with mycobacteria, macrophages secrete multiple cytokines and chemokines, including interleukin-6 (IL-6), IL-8 and tumor necrosis factor-α (TNF-α), to mediate host immune responses against the pathogen. Mycobacteria also induce the production of IL-10 <it>via </it>PKR activation in primary human monocytes and macrophages. As an anti-inflammatory cytokine, over-expression of IL-10 may contribute to mycobacterial evasion of the host immunity. <it>Radix Paeoniae Rubra </it>(RPR, <it>Chishao</it>), a Chinese medicinal herb with potentials of anti-inflammatory, hepatoprotective and neuroprotective effects, is used to treat tuberculosis. This study investigates the immunoregulatory effects of RPR on primary human blood macrophages (PBMac) during mycobacterial infection.</p> <p>Methods</p> <p>The interaction of Bacillus Calmette-Guerin (BCG) with PBMac was used as an experimental model. A series of procedures involving solvent extraction and fractionation were used to isolate bioactive constituents in RPR. RPR-EA-S1, a fraction with potent immunoregulatory effects was obtained with a bioactivity guided fractionation scheme. PBMac were treated with crude RPR extracts or RPR-EA-S1 before BCG stimulation. The expression levels of IL-6, IL-8, IL-10 and TNF-α were measured by qPCR and ELISA. Western blotting was used to determine the effects of RPR-EA-S1 on signaling kinases and transcriptional factors in the BCG-activated PBMac.</p> <p>Results</p> <p>In BCG-stimulated macrophages, crude RPR extracts and fraction RPR-EA-S1 specifically inhibited IL-10 production while enhanced IL-8 expression at both mRNA and protein levels without affecting the expressions of IL-6 and TNF-α. Inhibition of BCG-induced IL-10 expression by RPR-EA-S1 occurred in a dose- and time-dependent manner. RPR-EA-S1 did not affect the phosphorylation of cellular protein kinases including MAPK, Akt and GSK3β. Instead, it suppressed the degradation of IκBα in the cytoplasm and inhibited the translocation of transcription factor NF-κB1 p50 to the nucleus.</p> <p>Conclusion</p> <p>RPR crude extracts and its fraction RPR-EA-S1 inhibited anti-inflammatory cytokine IL-10 and enhanced pro-inflammatory chemokine IL-8 expression in BCG-activated PBMac. The inhibitory effects of RPR-EA-S1 on IL-10 expression in BCG-activated PBMac may be due to the reduced nuclear translocation of NF-κB1 p50.</p

Survival outcomes of a population-based cohort of adolescent and young adult (AYA), acute lymphoblastic leukemia (ALL) patients in Ontario, Canada: The IMPACT Cohort

Introduction Adolescents and young adults (AYA) are an understudied population in cancer research. The Initiative to Maximize Progress in Adolescent and Young Adult Cancer Therapy (IMPACT) cohort includes Ontario individuals aged 15-21 years, diagnosed with a malignancy during 1992-2011. This cohort contains a rich source of patient, disease, and treatment data. Objectives and Approach The IMPACT cohort was created using chart review and linkages to population-based health services databases. The cohort is comprised of AYA with a primary diagnosis of acute leukemia, Hodgkin lymphoma, non-Hodgkin lymphoma, sarcoma, or testicular cancer. For the current study, we focused on acute lymphoblastic leukemia (ALL) patients, and examined survival outcomes of those treated at pediatric vs adult centers, with adult vs pediatric protocols. We assessed 5-year event-free survival (first of relapse, progression, secondary malignant neoplasm, or death) and overall survival. Results The IMPACT cohort contains 2,963 patients, of which 152/271 ALL patients were treated at adult centers. The 5-year event-free survival (EFS ± standard error) among those treated at a pediatric vs. adult center was 72%±4% vs. 56%±4% (p = 0.03), respectively. The 5-year overall survival (OS) was 82%±4% vs. 64%±4% (p Conclusion/Implications ALL patients treated with a pediatric protocol at a pediatric center experienced better, crude survival outcomes compared to those at an adult center. The IMPACT cohort offers the opportunity to study a myriad of questions on different cancer groups, with the ultimate goal of improving outcomes among the AYA population

Mortality among single fathers as compared with single mothers and partnered fathers: a cohort study

Introduction Single-parent families are becoming increasingly common around the world with a particularly steep rise in households headed by single fathers. Research suggests that single parenthood is associated with adverse outcomes, however, little is known about the health profile or risk of death of single fathers compared to other parents. Objectives and Approach We aimed to examine mortality risk in single fathers compared to single mothers and partnered parents, which is the first head-to-head comparison of single and partnered parent groups. Our population consisted of 871 single fathers, 4590 single mothers, 16,341 partnered fathers and 18,688 partnered mothers. We combined cross-sectional samples derived from the Ontario component of Statistics Canada’s Canadian Community Health Survey (2001 to 2012) linked to health administrative databases to identify medical conditions, health service use, all-cause mortality and cause of death. We investigated differences in mortality using Cox proportional hazards models adjusted for socio-demographic characteristics, lifestyle and clinical factors. Results Each family group was followed up for a median of 11.10 years (IQR 7.36–13.54). Single fathers had a poor risk profile, particularly among lifestyle factors, such as low fruit and vegetable consumption and alcohol consumption. The mortality rate was three-fold higher for single fathers compared to single mothers and partnered fathers (5.81, 1.74 and 1.94, respectively). Even after adjustment, single fathers had a significantly higher risk of death compared to both single mothers (hazard ratio (HR): 2.49 (95% CI: 1.20 – 5.15, p=0.01)) and partnered fathers (HR: 2.06 (95% CI: 1.11 – 3.83, p=0.02). Small sample sizes precluded us from identifying the leading cause of death for single fathers. Conclusion/Implications We found that single fathers had the least favorable risk factor profile and greatest risk of mortality, which may be mitigated by physician intervention. Our study highlights that through cross-sectoral data linkages we can further advance our knowledge of social factors and their profound effects on health

Iodine-123 metaiodobenzylguanidine scintigraphic assessment of the transplanted human heart: Evidence for late reinnervation

Objectives.This study attempted to determine whether cardiac sympathetic reinnervation occurs late after orthotopic heart transplantation.Background.Metaiodobenzylguanidine (MIBG) is taken up by myocardial sympathetic nerves. Iodine-123 (I-123) MIBG cardiac uptake reflects intact myocardial sympathetic innervation of the heart. Cardiac transplant recipients do not demonstrate I-123 MIBG cardiac uptake when studied <6 months from transplantation. However, physiologic and biochemical studies suggest that sympathetic reinnervation of the heart can occur >1 year after transplantation.Methods.We performed serial cardiac I-123 MIBG imaging in 23 cardiac transplant recipients early (<-1 year) and late (>1 year) after operation. In 16 subjects transmyocardial norepinephrine release was measured late after transplantation.Results.No subject had visible I-123 MIBG uptake on imaging <1 year after transplantation. However, 11 (48%) of 23 subjects developed visible cardiac I-123 MIBG uptake 1 to 2 years after transplantation. Only 3 (25%) of 12 subjects with a pretransplantation diagnosis of idiopathic cardiomyopathy demonstrated I-123 MIBG uptake compared with 8 (73%) of 11 with a pretransplantation diagnosis of ischemic or rheumatic heart disease (p = 0.04). All 10 subjects with a net myocardial release of norepinephrine had cardiac I-123 MIBG uptake; all 6 subjects without a net release of norepinephrine had no cardiac I-123 MIBG uptake.Conclusions.Sympathetic reinnervation of the transplanted human heart can occur >1 year after operation, as assessed by I-123 MIBG imaging and the transmyocardial release of norepinephrine. Reinnervation is less likely to occur in patients with a pretransplantation diagnosis of idiopathic cardiomyopathy than in those with other etiologies of congestive heart failure

Contributions of Ethnicity to Differential Item Functioning of Cannabis Abuse and Dependence Symptoms

Cannabis is the most widely used illicit drug in the United States, and as a result, it is associated with significant public health costs. The present study sought to investigate whether item response theory (IRT) methods could be used to identify meaningful differences in how cannabis abuse and dependence symptoms (determined by criteria from the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) function as indices of the severity of misuse across two ethnic groups: Native Americans and European Americans

The Effects of Circumcision on the Penis Microbiome

Circumcision is associated with significant reductions in HIV, HSV-2 and HPV infections among men and significant reductions in bacterial vaginosis among their female partners.We assessed the penile (coronal sulci) microbiota in 12 HIV-negative Ugandan men before and after circumcision. Microbiota were characterized using sequence-tagged 16S rRNA gene pyrosequencing targeting the V3-V4 hypervariable regions. Taxonomic classification was performed using the RDP Naïve Bayesian Classifier. Among the 42 unique bacterial families identified, Pseudomonadaceae and Oxalobactericeae were the most abundant irrespective of circumcision status. Circumcision was associated with a significant change in the overall microbiota (PerMANOVA p = 0.007) and with a significant decrease in putative anaerobic bacterial families (Wilcoxon Signed-Rank test p = 0.014). Specifically, two families-Clostridiales Family XI (p = 0.006) and Prevotellaceae (p = 0.006)-were uniquely abundant before circumcision. Within these families we identified a number of anaerobic genera previously associated with bacterial vaginosis including: Anaerococcus spp., Finegoldia spp., Peptoniphilus spp., and Prevotella spp.The anoxic microenvironment of the subpreputial space may support pro-inflammatory anaerobes that can activate Langerhans cells to present HIV to CD4 cells in draining lymph nodes. Thus, the reduction in putative anaerobic bacteria after circumcision may play a role in protection from HIV and other sexually transmitted diseases

Inducible colitis-associated glycome capable of stimulating the proliferation of memory CD4+ T cells

Immune responses are modified by a diverse and abundant repertoire of carbohydrate structures on the cell surface, which is known as the glycome. In this study, we propose that a unique glycome that can be identified through the binding of galectin-4 is created on local, but not systemic, memory CD4+ T cells under diverse intestinal inflammatory conditions, but not in the healthy state. The colitis-associated glycome (CAG) represents an immature core 1–expressing O-glycan. Development of CAG may be mediated by down-regulation of the expression of core-2 β1,6-N-acetylglucosaminyltransferase (C2GnT) 1, a key enzyme responsible for the production of core-2 O-glycan branch through addition of N-acetylglucosamine (GlcNAc) to a core-1 O-glycan structure. Mechanistically, the CAG seems to contribute to super raft formation associated with the immunological synapse on colonic memory CD4+ T cells and to the consequent stabilization of protein kinase C θ activation, resulting in the stimulation of memory CD4+ T cell expansion in the inflamed intestine. Functionally, CAG-mediated CD4+ T cell expansion contributes to the exacerbation of T cell–mediated experimental intestinal inflammations. Therefore, the CAG may be an attractive therapeutic target to specifically suppress the expansion of effector memory CD4+ T cells in intestinal inflammation such as that seen in inflammatory bowel disease

Paradoxical risk perception and behaviours related to Avian Flu outbreak and education campaign, Laos

<p>Abstract</p> <p>Background</p> <p>In Laos, small backyard poultry systems predominate (90%). The first lethal human cases of highly pathogenic avian influenza (HPAI) occurred in 2007. Few studies have addressed the impact of outbreaks and education campaigns on a smallholder producer system. We evaluated awareness and behaviours related to educational campaigns and the 2007 HPAI outbreaks.</p> <p>Methods</p> <p>During a national 2-stage cross-sectional randomised survey we interviewed 1098 households using a pre-tested questionnaire in five provinces representative of the Southern to Northern strata of Laos. We used multivariate analysis (Stata, version 8; Stata Corporation, College Station, TX, USA) to analyse factors affecting recollection of HPAI educational messages, awareness of HPAI, and behaviour change.</p> <p>Results</p> <p>Of the 1098 participants, 303 (27.6%) received training on HPAI. The level of awareness was similar to that in 2006. The urban population considered risk to be decreased, yet unsafe behaviours persisted or increased. This contrasted with an increase in awareness and safe behaviour practices in rural areas.</p> <p>Reported behaviour changes in rural areas included higher rates of cessation of poultry consumption and dead poultry burial when compared to 2006. No participants reported poultry deaths to the authorities. Overall, 70% could recall an educational message but the content and accuracy differed widely depending on training exposure. Washing hands and other hygiene advice, messages given during the HPAI educational campaign, were not recalled. Trained persons were able to recall only one message while untrained participants recalled a broader range of messages. Factors associated with an awareness of a threat of AI in Laos were: having received HPAI training, literacy level, access to TV, recent information, living in rural areas.</p> <p>Conclusion</p> <p>We report a paradoxical relationship between unsafe behaviours and risk perception in urban areas, as well as exposure to HPAI training and message misinterpretation. Future educational campaigns need to be tailored to specific target populations and farming styles, for example, small holder farms as compared to commercial farms. Special attention must be given to varying risk perceptions and the risk of misinterpretation of key messages, economic hardship, and real life consequences of reporting.</p

Intercomparison of Field Methods for Acquiring Ground Reflectance at Railroad Valley Playa for Spectral Calibration of Satellite Data

Ground reflectance was acquired at the Railroad Valley Playa calibration site in Nevada USA using different methods of collection. The data was collected near the time and date of Landsat 8 OLI and Sentinel-2 satellite overpasses so an inter-comparison could be made with the reflectance products to determine which method was more suitable for vicarious calibration. The field spectrometers and reference panels were characterized before the field campaign. A continuous acquisition method was compared to stop and measure collections. Both acquisition methods were collected along an 80 m east-west transect as well as for a series of north-south transects over an 80 x 320 m area, with the stop and measure method being performed at random sampling locations. The measurements were performed using two field spectrometers by three teams of two people to compare the repeatability. The aim of the field campaign was to determine the variability due to the operator and the method of collection