The role of CD14 gene promoter polymorphism in tuberculosis susceptibility

BackgroundCD14 is expressed principally by cells of monocyte/macrophage lineage and plays a pivotal role in the innate immunity to intracellular infections. Recent research findings have revealed an association between the CD14 gene promoter polymorphism and several major infectious diseases.ObjectiveThe aim of the present study was to investigate the association between the CD14-159C/T polymorphism and tuberculosis in a Turkish population.MethodsFor this purpose, 88 consecutive patients with tuberculosis (63 pulmonary, 25 extrapulmonary) and 116 control subjects were enrolled into a prospective study. We determined CD14-159 genotypes by polymerase chain reaction - restriction fragment length polymorphism analysis and also measured serum concentrations of soluble CD14 (sCD14) by using a quantitative sandwich enzyme immunoassay technique.ResultsThere was no significant difference in terms of genotype distribution between patients with tuberculosis (CC 18.2%, CT 48.9%, TT 33.0%) and controls (CC 12.9%, CT 50.9%, TT 36.2%) or between patients with pulmonary and extrapulmonary tuberculosis. Serum levels of sCD14 were significantly increased in patients with active tuberculosis compared to those with inactive tuberculosis and healthy controls (p<0.001). However, levels of sCD14 were not associated with any genotypes of CD14-159.ConclusionThe genotyping findings of the present study do not support a role for the CD14-159C/T polymorphism in the development of tuberculosis, at least in the geographical region of central Anatolia. Significantly elevated serum sCD14 levels in patients with active disease reflect the importance of the mononuclear phagocytic system activation in tuberculosis

The role of SPINK5 in asthma related physiological events in the airway epithelium

SummaryBackgroundGenetic studies have shown that variants in SPINK5 may be associated with atopic diseases and asthma. However, the functional role of SPINK5 protein in asthma has not been elucidated.ObjectivesTo determine the effects of SPINK5 on asthma related physiological events such as apoptosis, mucus and cytokine production by epithelial cells.MethodsA549 cells were transfected with SPINK5 expression vector and stimulated with increasing doses of hydrogen peroxide and neutrophil elastase (NE) for measurement of cell viability or apoptosis and analysis of mucus production. Cell viability was measured by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyl-tetrazolium bromide) assay and apoptosis by Annexin V/PI staining. Levels of IL-4, IL-6, IL-8, IL-12, IL-13, IFNγ, IL-1β and RANTES were determined by ELISA in cell culture supernatants. Mucus production was determined by RT-PCR of the MUC5AC gene and PAS staining in NE treated cells.ResultsEpithelial cells transfected with SPINK5 expression vector produced more IL-6, IL-8 and RANTES compared to non-transfected cells (p < 0.001, p = 0.003, p < 0.001, respectively). Even though cells transfected with SPINK5 vector displayed significantly higher cell death, we have not observed any clear effect of SPINK5 on apoptosis. PAS staining showed that SPINK5 slightly decreased the mucin production induced by neutrophil elastase in A549 cells. However, SPINK5 had no effect on MUC5AC transcription.ConclusionSPINK5 is an important molecule in asthma. Its role extends beyond its well known protease inhibitor properties

The Role Of Scca1 In Asthma Related Physiological Events In The Airway Epithelium And The Effect Of Promoter Variants On Asthma And Gene Function

Background: Even though the systemic level of SCCA1, a serine protease inhibitor, was shown to be elevated in asthma, its physiological role is unknown. Objective: We sought to determine the effect of SCCA1 on apoptosis, cytokine expression and mucus production by A549 cells and define the effect of promoter variants on gene expression and association with asthma. Methods: SCCA levels were measured by ELISA. Promoter variants were determined by direct sequencing. 442 asthmatic children and 191 controls were genotyped by RFLP. The functional effect of the polymorphisms was assessed in transient transfection experiments using reporter constructs. A transcription factor ELISA was used for differential binding of GATA proteins to the variant region. The effects of SCCA1 on cytokine synthesis, mucus production and apoptosis were determined in A549 cells transfected with SCCA1 pcDNA vector. MUC5AC expression in A549 cells was determined with RT-PCR. Results: SCCA1 protein level was significantly higher in asthmatic children compared to healthy controls. Four polymorphisms SCCA1 promoter that were in linkage disequilibrium were associated with skin test positivity in asthmatic children and showed higher promoter activity and higher binding of GATA-2 and GATA-3 after IL-4 + IL-13 stimulation. IL-6, IL-8 levels were significantly higher in cells transfected with SCCA1 whereas RANTES increased only after IL-4 stimulation. Transfection of A549 cells with SCCA1 resulted in decreased MUC5AC expression and conferred protection against apoptosis. Conclusion: Our results showed that SCCA1 has diverse effects on many of the cellular events that characterize asthma and its role extends beyond protease inhibition. (C) 2012 Elsevier Ltd. All rights reserved.WoSScopu

A Potentially Fatal Outcome of Oral Contraceptive Therapy: Estrogen-Triggered Hereditary Angioedema in an Adolescent

Hereditary angioedema (HAE) is characterized by recurrent angioedema attacks with no urticaria. This disease has a high mortality due to asphyxia. Level of complement component 4 (C4), C1 esterase inhibitor (C1-INH) level and function, and genetic mutations determine different endotypes of HAE. Clinical presentation and the triggers of vasogenic edema may change according to the endotypes. An adolescent girl with oligomenorrhea, obesity, hirsutism, and acanthosis nigricans was diagnosed with polycystic ovary syndrome and prescribed ethinyl estradiol and cyproterone acetate containing oral contraceptive (OC). On the sixteenth day of treatment, she developed angioedema of the face, neck, and chest leading to dyspnea. Adrenaline, antihistamine, and corticosteroid treatments were ineffective. In the family history, the patient’s mother and two cousins had a history of angioedema. C1-INH concentrate was administered with a diagnosis of HAE. C4 and C1-INH level and activity were normal. Genetic analysis identified a mutation in the factor 12 (F12) gene, and the diagnosis of F12-related HAE was made. OC treatment was discontinued. She has had no additional angioedema attacks in the follow-up period of two years. OC containing estrogen may induce the life-threatening first attack of F12-related HAE even in children. Recurring angioedema attacks in the family should be asked before prescribing estrogen-containing OC pills

Pepsin levels and oxidative stress markers in exhaled breath condensate of patients with gastroesophageal reflux disease

KISA, Ucler/0000-0002-8131-6810; Soyer, Tutku/0000-0003-1505-6042WOS: 000327140800012PubMed: 24210194Aim: To evaluate the pepsin and oxidative stress markers in exhaled breath condensate (EBC) in patients with gastroesophageal reflux disease (GERD). Patients and Method: Patients with a presumptive diagnosis of GERD with recurrent respiratory and gastrointestinal problems aged between 2 and 14 years were included in the study. All patients underwent pH monitoring. Patients with a reflux index (RI) >= 4 were assessed as the reflux group, and those with an RO 0.05)]. Conclusion: Decreased levels of NOX in patients with GER disease suggest increased oxidative stress in airways of these patients. (C) 2013 Elsevier Inc. All rights reserved.Kirikkale University Scientific Research UnitKirikkale UniversityThis study was supported by Kirikkale University Scientific Research Unit and presented in National Congress of Turkish Pediatric Surgeons on 30 September-3 October 2009, in Malatya, Turkey

Monocyte chemoattractant protein-4 core promoter genetic variants: influence on YY-1 affinity and plasma levels.

Monocyte chemoattractant protein-4 (MCP-4) is a CC chemokine implicated in the recruitment of eosinophils, monocytes, and T-lymphocytes in diseases of mucosal inflammation, including asthma. We tested the hypothesis that there is a genetic basis for differences in MCP-4 expression among individuals by evaluating the effects of core promoter variants on MCP-4 expression. We identified two single-nucleotide T-to-C polymorphisms in the MCP-4 core promoter that occur 896 and 887 base pairs preceding the transcription initiation site. The -887 variant alters a consensus binding motif for the transcription factor YY-1. Electrophoretic mobility shift assay demonstrated that YY-1 containing nuclear extracts from tumor necrosis factor-alpha-stimulated peripheral blood mononuclear cells had greater avidity for the wild-type (YY-1 motif intact) sequence than for the variant sequence. Increasing doses of a YY-1 expression vector induced significantly greater reporter activity from MCP-4 core promoter expression constructs of the wild-type compared with the variant sequence in transient transfection experiments. The external validity of these observations was demonstrated by measuring plasma levels of MCP-4 from individuals with the alternative forms of the gene. Individuals bearing haplotypic variants of the MCP-4 core promoter that avidly bind the transcription factor YY-1 had higher plasma levels of MCP-4 than did individuals with variants with lower binding avidity (490, 360, and 360 pg/ml; P \u3c 0.01). Our findings suggest that the MCP-4 core promoter YY-1 binding motif is functional, modulates the transcriptional regulation of the MCP-4 gene, and that part of the variance in the systemic expression of MCP-4 is determined by core promoter genetic variants