Common phenotype and different non-HLA genes in Graves’ disease and alopecia areata

Our previous observations clarified that Graves’ disease (GD) is the most frequent autoimmune disease in patients with alopecia areata (AA), and 42.7% of patients with AA were positive for thyrotropin receptor antibody (TRAb). A class II HLA haplotype DRB1¹15:01-DQB1¹06:02 was suggested to contribute to autoimmunity against the thyroid gland in AA. To further clarify the genetic factors contributing to organ specificity in autoimmune diseases, we studied the contribution of non-HLA genes to organ specificity in GD and AA. A high frequency of AA (13.4%) was observed in patients with GD, indicating strong phenotypic association between GD and AA. CTLA4 and TSHR were significantly associated with GD (Pc = 0.007 and Pc < 0.002, respectively), but not with AA, even in TRAb-positive patients. The difference in the association between GD and AA suggests that the CTLA4 and TSHR are not main factors contributing to determining common genetic basis among GD and AA.     

MicroRNA–21 as a novel biomarker in diagnosis and response to therapy in asthmatic children

BackgroundThe underlying molecular mechanisms leading to asthma remain largely unclear. MicroRNAs (miRNAs) are short noncoding RNAs exert powerful effects on immunological function by tuning networks of target genes that orchestrate cell activity. However, the role of miRNAs, specifically microRNA-21 (miRNA- 21), in the regulation of allergic airway inflammation is not well defined. Our aim was to investigate the serum miRNA- 21 expression levels as potential biomarker in childhood asthma [with, without inhaled corticosteroid (ICS) therapy, and steroid resistant (SR)]; and their possible contributions in disease status, its molecular target interleukin-12 (IL-12) p35, and response to therapy.Materials and methodsThis study included 175 children; 95 were asthmatic patients subdivided into 3 groups [40 asthmatic children without ICS, 40 steroid sensitive (SS) asthma children and 15 steroid resistant (SR) asthma children] and 80 were healthy children as healthy controls. The miRNA-21 expressions levels were determined by quantitative real-time polymerase chain reaction (qRT-PCR) in all children. Serum IL-12p35 and total IgE levels were measured using enzyme-linked immunosorbent assay (ELISA).ResultsThe expression levels of miRNA-21 were significantly higher in the asthmatic children than in control group (P < 0.001); with significantly higher levels in asthmatic patients without ICS or in SR patients compared to SS children (P < 0.001). On contrast, serum IL-12p35 levels were significantly decreased in asthmatic patients without ICS therapy or in SR asthma patients as compared to SS patients (P < 0.001). Our data revealed that serum miRNA-21 expression levels was significant negatively correlated with serum IL-12p35 levels and FEV1, while it was positively correlated with both sputum and blood eosinophils. Importantly, serum miRNA-21 had a predictive value in differentiating SS from SR patients, with an AUC value of 0.99, specificity of 86.7%, sensitivity of 97.5% and P < 0.001.ConclusionThis study suggested that serum miRNA-21 is stable and detectable in serum of asthmatic children, which could promise potential biomarker in diagnosis as well as in response to therapy of asthma.     

Chlamydia pneumoniae enhances the Th2 profile of stimulated peripheral blood mononuclear cells from asthmatic patients

Chlamydia pneumoniae is a cause of respiratory infection in adults and children. There is evidence for an association between atypical bacterial respiratory pathogens and the pathogenesis of asthma. We compared T helper (Th) responses in C. pneumoniae – infected peripheral blood mononuclear cells (PBMC) in patients with or without asthma. PBMC (1 × 10⁶/mL) from asthmatic patients (N = 11) and non-asthmatic controls (N = 12) were infected or mock-infected for 1 h +/− C. pneumoniae TW-183 at a multiplicity of infection (MOI) = 1 and MOI = 0.1, or cultured for 24 h +/− Lactobacillus rhamnosus GG (LGG). Interleukin (IL)-4, IL-10, IL-12, Interferon (IFN)-gamma and total IgE levels were measured in supernatants (ELISA). C. pneumoniae infection led to an increase (>50%) of IgE levels in PBMC from asthmatics, compared with mock-infected on day 10; IgE wasn’t detected in non-asthmatics. C. pneumoniae – infected PBMC from asthmatics increased levels of IL-4 and IFN-gamma after 24 h, compared with PBMC alone; levels of IL-10 and IL-12 were low. When uninfected-PBMC from asthmatics were LGG-stimulated, after 24 h, IL-4 was undetectable, but IL-10, IL-12, and IFN-gamma increased, compared with PBMC alone. Thus, C. pneumoniae infection has the ability to induce allergic responses in PBMC of asthmatics, as evidenced by production of Th2 responses and IgE.     

Elevated neopterin levels in non-allergic asthma

Neopterin is synthesized by human monocyte-derived macrophages upon stimulation with interferon-γ (IFN-γ). Measurement of neopterin concentration is useful to monitor cell-mediated (Th1-type) immune activation.In this study, we aimed to analyze the behaviour of neopterin in long lasting asthma considering its role as a marker of the Th1 environment and to establish the distinction between patients belonging either to the allergic or the non-allergic population, particularly in the elderly where asthma is often under diagnosed. Therefore we evaluated allergic parameters such as skin prick tests, IgE and hemogram (eosinophils count), and we compared our findings with neopterin values found in an age-matched control population.A group of individuals older than 65 was selected. It included 64 asthmatic patients (mean age 72 ± 5 years) and 41 healthy individuals (mean age 79 ± 7 years). In our study population, 42 patients presented positive skin tests, mainly to house dust mites. All patients were clinically stable and presented an average percentage of predicted forced expiratory volume in the first second (FEV1) of 73.6 ± 25.3 and predicted median expiratory flow percentage (MEF50) of 38.8 ± 26.7.Blood cell counts showed statistically different mean values of eosinophils between allergic and non-allergic controls (5.42 ± 4.7% versus 2.8 ± 2.8%; p < 0.04). IgE values were increased in allergic asthmatic patients when compared with non-allergic asthmatic patients (493.2 ± 549.8 IU/ml versus 85.3 ± 194.4 IU/ml; p = 0.000).Allergic asthmatic patients presented mean neopterin levels similar to those found in the control group (2.4 ± 2.8 ng/ml versus 2.1 ± 1.9 ng/ml). In contrast, in non-allergic asthmatic patients these values were higher when compared with the control group (4.0 ± 4.7 ng/ml versus 2.1 ± 1.9 ng/ml). Neopterin levels were lower in allergic asthmatic patients when compared with non-allergic asthmatic patients (2.4 ± 2.8 ng/ml versus 4.0 ± 4.7 ng/ml).Within asthmatic patients, those with higher neopterin values (>2.1 ng/ml) presented lower mean IgE values (IgE ≤ 336.58 IU) than those with lower neopterin values (≤2.1 ng/ml) who presented mean IgE values of 402.70 IU.Our initial findings may lead to a better understanding of the immunoinflammatory pathways in asthma. Further studies will probably show that serum neopterin could became a useful marker for asthma classification including in elderly patients with long lasting disease.     

IgE anti-respiratory syncytial virus antibodies detected in serum of pediatric patients with asthma

Respiratory syncytial virus (RSV) causes lower respiratory tract disease in infants and young children, and is a public health concern, as is the increase in pediatric asthma. Respiratory viral infections may trigger asthma exacerbations. However, it remains unknown whether RSV infection may have a specific association with asthma. Total serum IgE, and IgE- and IgG-anti-RSV Ab responses were studied in older asthmatic compared with non-asthmatic children (M/F, mean age: 14) (N = 30, N = 43, respectively). We found: (1) total serum IgE was higher in asthmatic compared with non-asthmatics (P < 0.001); (2) total serum IgE did correlate with IgE anti-RSV Abs (P < 0.001), and with IgG anti-RSV Abs (P = 0.008) in all subjects; (3) total serum IgE levels did correlate with IgE anti-RSV in asthmatics (P = 0.047), but not in non-asthmatics (P = 0.13); (4) IgE anti-RSV Abs did correlate with IgG anti-RSV Abs in all subjects (P = 0.001); (5) IgE- and IgG-anti RSV Abs were higher in asthma compared with no asthma (P = 0.003; <0.001, respectively); (6) there was a significant association between age and IgE anti-RSV in non-asthma (P = 0.008), but not in asthma (P = 0.64). Our findings indicate that IgE-anti-RSV Ab responses may play important roles in RSV infection and asthma.     

Macrophage migration inhibitory factor contributes to anti-neutrophil cytoplasmic antibody-induced neutrophils activation

BackgroundMacrophage migration inhibitory factor (MIF) is an inflammatory mediator released by macrophages that is central to the innate immune system, with an upstream role in the inflammatory cascade. MIF is one of the most important pathogenic factors in the development of the autoimmune diseases. In the current study, we investigated the role of MIF in anti-neutrophil cytoplasmic antibody (ANCA)-induced neutrophil activation.MethodsPlasma levels of MIF from 31 patients with active ANCA-associated vasculitis (AAV) were analyzed by ELISA. The various effects of MIF in ANCA-induced neutrophil respiratory burst and degranulation were measured.ResultsPlasma levels of circulating MIF were significantly higher in AAV patients with active disease compared with those in remission and healthy controls. Compared with MIF-primed neutrophils, the MFI value increased significantly in MIF-primed neutrophils further activated with MPO-ANCA-positive IgG or PR3-ANCA-positive IgG (270.8 ± 9.7 vs. 421.5 ± 9.7, P < 0.001; 270.8 ± 9.7 vs. 414.1 ± 15.6, P < 0.001, respectively). Compared with MIF-primed neutrophils, the lactoferrin concentration increased significantly in the supernatant of MIF-primed neutrophils further activated by MPO-ANCA-positive IgG (567.8 ± 61.2 ng/ml vs. 1677.0 ± 42.5 ng/ml, P < 0.001) or PR3-ANCA-positive IgG (567.8 ± 61.2 ng/ml vs. 1546.0 ± 116.2 ng/ml, P < 0.001), respectively. Interleukin-8 (IL-8), IL-6 and IL-23 were involved in ANCA-induced activation of MIF-primed neutrophils.ConclusionsMIF primes neutrophils by increasing ANCA antigen translocation. The primed neutrophils can be further induced by ANCA, resulting in respiratory burst and degranulation.     

Dysregulated production of interleukin-1β upon activation of the NLRP3 inflammasome in patients with familial Mediterranean fever

Familial Mediterranean fever (FMF) is caused by mutations in pyrin, a protein expressed in innate immune cells that interacts with caspase-1 and other inflammasome components to regulate interleukin (IL)-1β maturation. Since NLRP3 inflammasome represents major source of IL-1β, we studied its protein expression and function in FMF. We isolated peripheral white blood cells (WBCs) from 20 symptoms-free FMF patients and 21 healthy individuals. Intracellular protein expression of NLRP3, caspase-1, IL-1β at baseline and after LPS/ATP sequential treatment for NLRP3 activation was assessed by immunoblotting. Secreted IL-1β was quantified by ELISA. THP-1 cells were transfected with wild-type or mutant pyrin and IL-1β secretion was measured. FMF WBCs exhibited lower NLRP3 and active caspase-1 protein expression compared to healthy individuals, and LPS/ATP treatment resulted in significantly lower intracellular IL-1β levels in FMF patients. Likewise, LPS/ATP induced caspase-1-dependent IL-1β release at significantly lower amounts in the FMF group (1182 ± 192 versus 2134 ± 245 pg/mL in controls, p = 0.004). Consistently, THP-1 cells transfected with FMF-associated M694V mutant pyrin displayed lower LPS/ATP-induced IL-1β compared with wild-type pyrin-transfected cells. FMF WBCs demonstrate reduced NLRP3-mediated IL-1β production. Additional studies are needed to define whether this finding represents a compensatory mechanism to control inflammation or is directly linked to disease pathogenesis.     

Association of IL-17A and IL-17F polymorphisms with gastric cancer risk in Asians: A meta-analysis

Increasing number of studies focused on the association of IL-17A rs2275913 and IL-17F rs763780 polymorphisms with gastric cancer (GC) risk. However, the results were inconsistent. To elucidate the exact association, we performed the present meta-analysis. Databases including PubMed, Web of knowledge and Chinese National Knowledge Infrastructure (CNKI) were systematically searched for potentially eligible literatures. Odds ratios (OR) and their 95% confidence interval (CI) were used to evaluate the strength of association. Eight studies for IL-17A rs2275913 (3345 cases and 4427 controls) and five studies for IL-17F rs763780 (1784 cases and 2592 controls) were finally included. The results indicated that individuals with AA genotype of IL-17A rs2275913 polymorphism were associated with increased GC risk compared with wild-type GG (OR = 1.61, 95% CI = 1.17–2.23, P = 0.004); A allele was significantly associated with increased GC risk compared with G allele (OR = 1.22, 95% CI = 1.06–1.41, P = 0.007). IL-17F rs763780 polymorphism was also significantly associated with increased GC risk (CC vs. CT: OR = 1.40, 95% CI = 1.04–1.88, P = 0.025; CT vs. TT: OR = 1.35, 95% CI = 1.16–1.58, P < 0.001; C allele vs. T allele: OR = 1.30, 95% CI = 1.15–1.47, P < 0.001). In summary, IL-17A rs2275913 A/G polymorphism and IL-17F rs763780 C/T polymorphism might be associated with increased GC risk in Asians. Further large-scale studies are still required to confirm the results of this meta-analysis.     

Implementation and evaluation of an integrated computerized asthma management system in a pediatric emergency department: A randomized clinical trial

ObjectiveThe use of evidence-based guidelines can improve the care for asthma patients. We implemented a computerized asthma management system in a pediatric emergency department (ED) to integrate national guidelines. Our objective was to determine whether patient eligibility identification by a probabilistic disease detection system (Bayesian network) combined with an asthma management system embedded in the workflow decreases time to disposition decision.MethodsWe performed a prospective, randomized controlled trial in an urban, tertiary care pediatric ED. All patients 2–18 years of age presenting to the ED between October 2010 and February 2011 were screened for inclusion by the disease detection system. Patients identified to have an asthma exacerbation were randomized to intervention or control. For intervention patients, asthma management was computer-driven and workflow-integrated including computer-based asthma scoring in triage, and time-driven display of asthma-related reminders for re-scoring on the electronic patient status board combined with guideline-compliant order sets. Control patients received standard asthma management. The primary outcome measure was the time from triage to disposition decision.ResultsThe Bayesian network identified 1339 patients with asthma exacerbations, of which 788 had an asthma diagnosis determined by an ED physician-established reference standard (positive predictive value 69.9%). The median time to disposition decision did not differ among the intervention (228 min; IQR = (141, 326)) and control group (223 min; IQR = (129, 316)); (p = 0.362). The hospital admission rate was unchanged between intervention (25%) and control groups (26%); (p = 0.867). ED length of stay did not differ among intervention (262 min; IQR = (165, 410)) and control group (247 min; IQR = (163, 379)); (p = 0.818).ConclusionsThe control and intervention groups were similar in regards to time to disposition; the computerized management system did not add additional wait time. The time to disposition decision did not change; however the management system integrated several different information systems to support clinicians’ communication.     

Evidence of association of interleukin-23 receptor gene polymorphisms with Egyptian rheumatoid arthritis patients

BackgroundThe identification of additional genetic risk factor is an on-going process that will aid in the understanding of rheumatoid arthritis (RA) aetiology. A genome-wide association scan in Crohn’s (CD) disease highlighted the interleukin-23 receptor (IL23R) gene as a susceptibility factor. Since the IL-23/IL-17 pathway is known to associate with other autoimmune disease, including rheumatoid arthritis and systemic sclerosis, we hypothesised that IL23R could be a shared susceptibility gene. The rare allele of IL23R single nucleotide polymorphism (SNP) rs11209026 (Arg381Gln) confers strong protection against CD. Our aim was to analyse IL23R SNP (rs11209026, rs2201841, and rs10889677) and to detect its association with RA in Egyptian patients.MethodsA group of Egyptian patients with RA (n = 120) and apparently healthy persons as controls (n = 120) was genotyped for rs11209026, rs2201841 and rs10889677 by real time/polymerase chain reaction (real-time/PCR) for the first SNP and restriction fragment length polymorphism/PCR (RFLP/PCR) in the last two SNPs.ResultsOur data emphasise that the AA genotype of rs11209026 (Arg381Gln) was significantly associated with RA patients compared to the controls (P value = 0.001).We did not find any significant association between either rs2201841 or rs10889677 and the development of rheumatoid arthritis (P value = 1.000 & 0.562 respectively).ConclusionOur results suggest that IL23 receptor AA genotype variant of rs11209026 would contribute to RA aetiology; consequently, it might be a genetic marker for RA. We need to address the subgroup of patients who will benefit from the selective suppression of the IL23 signalling which would represent new perspectives toward a personalized therapy of RA patients by further studies.     

Maturation of human iDCs by IL-18 plus PGE2, but not by each stimulus alone,induced migration toward CCL21 and the secretion of IL-12 and IFN-γ

Dendritic cells (DCs) are potent antigen-presenting cells that initiate the primary immune response and whose functional properties in vivo depend on the maturation stimulus. We describe the functional properties of human monocyte-derived DCs after the maturation of immature DCs (iDCs) for 2 days with LPS (100 ng/ml), PGE2 (1 μg/ml), CD40L (1 μg/ml) or IL-18 (200 ng/ml) and with CD40L + PGE2 and IL-18 + PGE2 mixtures at the same concentrations as above. Neither IL-18 nor PGE2 alone stimulated IL-12 or IFN-γ secretion. When administered simultaneously to 1 × 10⁶ iDCs/ml, IL-18 + PGE2 induced the secretion of 131.4 ± 6.7 pg IL-12/ml and 355 ± 87 pg IFN-γ/ml but there was no detectable IL-10 secretion. However, PGE2 alone stimulated the secretion of 208 ± 89 pg IL-10/ml whereas IL-18 alone did not stimulate the secretion of IL-10, IL-12, TNF-α or INF-γ. When the mixture of CD40L + PGE2 was used, only migration toward CCL19 and CCL21 was induced. CD40L did not stimulate the secretion of IL-10, IL-12, TNF-α or IFN-γ and did not stimulate migration toward CCL19 or CCL21. The extent of stimulation of T cell proliferation was essentially the same for all stimuli at the concentrations given above. New properties such as IL-12 and INF-γ secretion and migration toward CCL21 emerged when a mixture of IL-18 + PGE2 was employed. These data show that when the pairs of stimuli reported here were used simultaneously their effect was not additive. This system can be used to prepare mDCs with properties useful for cell therapy and also as a model to investigate the mechanisms of cytokine secretion and cell migration.     

Genetic polymorphisms in TNFSF13 and FDX1 are associated with IgA nephropathy in the Han Chinese population

IgA nephropathy (IgAN) is the most common primary glomerulonephritis worldwide, and its pathogenesis is influenced by both genetic and environmental factors. In this study, we evaluated 23 tag single-nucleotide polymorphisms (tSNPs) in 21 IgAN-associated genes, in 200 subjects with IgAN and 310 healthy gender- and age-matched unrelated control subjects with no history of renal disease or hypertension. Using the co-dominant model, we found that two genotypes of rs3803800 in TNFSF13 were associated with an increased risk of IgAN: “GA” (OR = 1.03, 95% CI = 0.71–1.51, p = 0.018) and “AA” (OR = 2.45, 95% CI = 1.29–4.65, p = 0.018). The “AA” genotype was also associated with an increased risk of IgAN in the recessive model (OR = 2.41, 95% CI = 1.30–4.46, p = 0.018), as was the genotype “AA” rs10488764 in FDX1 (OR = 1.88, 95% CI = 1.01–3.53, p = 0.048). Interestingly, we found that the allele “A” of rs3803800 in TNFSF13 is associated with a decreased risk of IgAN in females (OR = 0.43, 95% CI = 0.20–0.95, p = 0.009), but with an increased risk in males (OR = 1.78, 95% CI = 0.86–3.66, p = 0.009). Our findings, combined with previously reported results, suggest that TNFSF13 and FDX1 have potential roles in IgAN in the Han Chinese population. This information may be useful in the development of early prognostics for IgAN.     

An asthma management system in a pediatric emergency department

IntroductionPediatric asthma exacerbations account for >1.8 million emergency department (ED) visits annually. Asthma guidelines are intended to guide time-dependent treatment decisions that improve clinical outcomes; however, guideline adherence is inadequate. We examined whether an automatic disease detection system increases clinicians’ use of paper-based guidelines and decreases time to a disposition decision.MethodsWe evaluated a computerized asthma detection system that triggered NHLBI-adopted, evidence-based practice to improve care in an urban, tertiary care pediatric ED in a 3-month (7/09–9/09) prospective, randomized controlled trial. A probabilistic system screened all ED patients for acute asthma. For intervention patients, the system generated the asthma protocol at triage for intervention patients to guide early treatment initiation, while clinicians followed standard processes for control patients. The primary outcome measures included time to patient disposition.ResultsThe system identified 1100 patients with asthma exacerbations, of which 704 had a final asthma diagnosis determined by a physician-established reference standard. The positive predictive value for the probabilistic system was 65%. The median time to disposition decision did not differ among the intervention (289 min; IQR = (184, 375)) and control group (288 min; IQR = (185, 375)) (p = 0.21). The hospital admission rate was unchanged between intervention (37%) and control groups (35%) (p = 0.545). ED length of stay did not differ among the intervention (331 min; IQR = (226, 581)) and control group (331 min; IQR = (222, 516)) (p = 0.568).ConclusionDespite a high level of support from the ED leadership and staff, a focused education effort, and implementation of an automated disease detection, the use of the paper-based asthma protocol remained low and time to patient disposition did not change.     

Regulatory role of Cdx-2 and Taq I polymorphism of vitamin D receptor gene on chemokine expression in pulmonary tuberculosis

Vitamin D receptor (VDR) gene variants have been shown to be regulating the immune response in tuberculosis. We studied the regulatory role of VDR promoter Cdx-2 and 3′UTR TaqI gene variants on chemokine levels from culture filtrate antigen (CFA) stimulated with or without 1,25(OH)₂D₃ treated peripheral blood mononuclear cells of 50 pulmonary tuberculosis patients (PTB) and 51 normal healthy controls (HCs). In CFA with 1,25(OH)₂D₃ treated cultures, the MIP-1α, MIP-1β, RANTES levels were significantly decreased in Cdx-2 AA genotype compared to GG genotype, while a significantly increased MIG level was observed in Cdx-2 AA genotype (p < 0.05). In TaqI polymorphism, tt genotype significantly decreased MIP-1β and RANTES levels compared to TT genotype. Moreover, a significantly increased level of IP-10 and MIG was observed in TaqI tt genotype compared with TT genotype (p < 0.05). The results suggests that the 1,25(OH)₂D₃ may alter the chemokine response through the VDR polymorphic variants during infection.     

Breastfeeding and IL-10 levels in children affected by cow’s milk protein allergy: A restrospective study

AimTo assess the protective role of breast-feeding in infants with CMPA-related AEDS as well as IL-10 utility as marker of disease evolution.Methods64 breast-feeding children with CMPA-related AEDS (31 males and 33 females; mean age 5.56 ± 2.41 months; 21 mild AEDS; 25 moderate AEDS; 18 severe AEDS) and 60 artificial feeding babies (33 males and 27 females; mean age 6.01 ± 2.08 months; 26 mild AEDS; 19 moderate AEDS; 15 severe AEDS) were evaluated. In all patients serum IL-10 levels were detected.ResultsSignificant Score Atopic Dermatitis (SCORAD) index point differences between breastfed and not breastfed children (p < 0.001) have been detected. The serum IL-10 levels were lower in children with CMPA-related AEDS as compared to the healthy control group (p < 0.001). Moreover, a significant inverse correlation between serum IL-10 levels and SCORAD in both enrolled groups has been also noted. In particular, IL-10 levels, in both groups, were significantly lower in children with severe symptoms. Conversely, serum IL-10 levels were significantly increased in children with mild-severe symptoms in both groups. Furthermore, breastfed children, with lower severe symptoms, had higher serum IL-10 levels. Finally, serum total IgE levels were negatively correlated with serum IL-10 levels in both breastfed and non-breastfed children with CMPA-related AEDS (p < 0.001).ConclusionsWe reported that exclusive breast-feeding induces hyposensitization in children with CMPA-related AEDS and it is associated with minor disease severity and higher serum IL-10 levels, resulting as useful disease-monitor marker.     

Urban air pollution produces up-regulation of myocardial inflammatory genes and dark chocolate provides cardioprotection

Air pollution is a serious environmental problem. Elderly subjects show increased cardiac morbidity and mortality associated with air pollution exposure. Mexico City (MC) residents are chronically exposed to high concentrations of fine particulate matter (PM_2.5) and PM-associated lipopolysaccharides (PM-LPS). To test the hypothesis that chronic exposure to urban pollution produces myocardial inflammation, female Balb-c mice age 4 weeks were exposed for 16 months to two distinctly different polluted areas within MC: southwest (SW) and northwest (NW). SW mice were given either no treatment or chocolate 2 g/9.5 mg polyphenols/3 times per week. Results were compared to mice kept in clean air. Key inflammatory mediator genes: cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), and the LPS receptor CD14 (cluster of differentiation antigen 14) were measured by real-time polymerase chain reaction. Also explored were target NFκB (nuclear factor κB), oxidative stress and antioxidant defense genes.TNF-α, IL-6, and COX-2 were significantly increased in both NW and SWMC mice (p = 0.0001). CD14 was up-regulated in SW mice in keeping with the high exposures to particulate matter associated endotoxin. Chocolate administration resulted in a significant down-regulation of TNF-α (p < 0.0001), IL-6 (p = 0.01), and IL-1β (p = 0.02). The up-regulation of antioxidant enzymes and the down-regulation of potent oxidases, toll-like receptors, and pro-apoptotic signaling genes completed the protective profile.Exposure to air pollution produces up-regulation of inflammatory myocardial genes and endotoxin plays a key role in the inflammatory response. Regular consumption of dark chocolate may reduce myocardial inflammation and have cardioprotective properties in the setting of air pollution exposures.     

Polymorphisms of IFN-γ (+874A/T) and IL-12 (+1188A/C) in tuberculosis patients and their household contacts in Hyderabad,India

Several cytokine gene variants have shown to be associated with host susceptibility to infectious diseases including tuberculosis (TB). High rates of transmission were identified within household members of TB patients. In this study, we examined whether single nucleotide polymorphisms of IFN-γ +874A/T and IL-12 +1188A/C affect susceptibility to TB. Genomic DNA from patients with active disease, their household contacts HHC and healthy controls HC was genotyped for IFN-γ +874A/T and IL-12 +1188A/C SNPs by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). IFN-γ +874 AA and AT genotypes were significantly with different frequencies in patients and total HHC as compared to HC (p < 0.0001). In patients IL-12 +1188 AC and CC genotypes were associated with TB (p < 0.003, p < 0.008). In total HHC AC, CC genotypes and both alleles (A&C) were significantly different as compared to HC (p < 0.004, p < 0.001, p < 0.034) and the same result was obtained when HHC were stratified into related (p < 0.02, p < 0.001) and unrelated (p < 0.009, p < 0.017) individuals. Allelic frequencies, however, were significant only in related contacts (p < 0.021). Generalized multifactor dimensionality reduction method (GMDR) testing revealed high risk combinations of several genotypes in IFN-γ & IL-12 genes. Our findings suggest an important role of genetic variations of IFN-γ and IL-12 for susceptibility to TB.     

Involvement of genes encoding apoptosis regulatory factors (FAS,FASL, TRAIL,BCL2, TNFR1 and TNFR2) in the pathogenesis of autoimmune thyroid diseases

Apoptosis is necessary for the maintenance of self-tolerance by eliminating autoreactive immune cells in the periphery. To clarify the association between the pathogenesis of autoimmune thyroid disease (AITD) and genes encoding apoptosis regulatory factors, we genotyped the FAS −1377G/A, −670A/G, FASL −844C/T, TRAIL −716C/T, BCL2 −938C/A, +127G/A, TNFR1 −383A/C and TNFR2 +676T/G polymorphisms. The frequencies of the FASL −844CC and BCL2 −938AA genotypes were significantly lower in AITD patients than in control subjects (P = 0.0101 and 0.0307, respectively). The frequency of the TNFR2 +676TT genotype was significantly lower in Graves’ disease (GD) patients than in controls (P = 0.0284). The serum sFasL level was significantly higher in GD and Hashimoto’s disease (HD) patients than in control subjects (P = 0.0003 and 0.0017, respectively). The serum sFasL levels in control subjects were significantly lower than those in intractable GD, GD in remission, and HD without treatment (P = 0.0310, 0.0007 and 0.0002, respectively). The serum sFasL levels in HD with treatment were significantly lower than those in HD without treatment (P = 0.0490). The polymorphisms in genes encoding apoptosis regulatory factors (FASL, BCL2) and serum levels of sFasL may be associated with immune dysregulation.     

Bone mineral density in pediatric survivors of Hodgkin and non-Hodgkin lymphomas

PurposeTo assess skeletal mass in survivors of childhood Hodgkin disease (HD) and non-Hodgkin lymphoma (NHL) 1–5 years after treatment, and to identify potential risk factors influencing bone mineral density (BMD).Patients/methodsThis cross-sectional study was conducted in a cohort of 43 survivors (HD = 31; NHL = 12); mean age: 16.21 ± 4.4. Total body bone mineral content (TBMC) and density (TBBMD), and lumbar spine density (LSBMD) were determined using dual-energy X-ray absorptiometry.ResultsThree of all 43 patients developed low BMD. No significant differences in height, weight, and/or BMD Z-scores were found between HD and NHL survivors, children who received and did not receive radiotherapy, and the groups with different chemotherapeutic blocks. No differences were noted between the Z-scores of BMC (mean ± SD: 0.31 ± 1.29 vs. −0.089 ± 0.61, p = 0.165), TBBMD (mean ± SD: −0.32 ± 1.21 vs. −0.27 ± 0.91, p = 0.76), or the LSBMD (mean ± SD: −0.183 ± 1.54 vs. −0.17 ± 0.87, p = 0.637) in subgroups, in accordance with time after therapy (subgroup I < 2 years and subgroup II > 2 years after treatment). In HD survivors, age at diagnosis only affected the TBBMD Z-score (a decrease of 0.127 in total BMD Z-score per each year, R² = 0.999, p < 0.001).ConclusionsChildhood lymphoma survivors demonstrate no significant deficits in bone mass and tend to maintain their BMD within the normal range when presenting during one to five years’ follow-up. However, this specific group requires longitudinal investigation to assess the pattern of peak bone mass achievement and the risk of future bone loss.