DNA (cytosine-5)-methyltransferase 3B (DNMT 3B) polymorphism and risk of Down syndrome offspring

Down syndrome (DS) is the most common form of human genetic mental retardation. Several polymorphisms in genes coding folic acid cycle enzymes have been associated to the risk of bearing a DS child; however, the results are controversial. S-adenosyl-l-methionine (SAM) is an important intermediate of folic acid pathway and acts as methyl donor and substrate for DNA (cytosine-5)-methyltransferase 3B (DNMT3B – EC 2.1.1.37) de novo methylation processes during embryogenesis. Recent studies suggest that a functional polymorphism of DNMT 3B in maternal genotype may be associated with a decreased risk of having a DS child. We herein investigate the association of this polymorphism with the occurrence of DS in a Brazilian population. We have genotyped 111 mothers of DS infants (MDS) and 212 control mothers (CM) through PCR-RFLP. The observed genotypic frequencies were CC = 0.22; CT = 0.49 and TT = 0.29 in CM, and CC = 0.30; CT = 0.52 and TT = 0.18 in MDS. Allelic frequencies were C = 0.47 and T = 0.53 in CM and C = 0.56 and T = 0.44 in MDS. No deviation of HWE was observed, and both DNMT 3B rs2424913 genotype (χ2 = 4.53; DF = 1; P = 0.03) and allelic (χ2 = 4.90; DF = 1; P = 0.03) frequencies show significant differences between MDS and CM. The presence of the mutant DNMT 3B T allele decreases 30% the risk of bearing a DS child (OR = 0.69; 95% CI: 0.50–0.96; P = 0.03), and the risk is diminished up to 45% in association with the homozygous genotype (OR = 0.54; 95% CI: 0.31–0.96; P = 0.04). Our results suggest that women harboring the single nucleotide polymorphism DNMT 3B rs2424913 have a decreased risk of a DS pregnancy, and further studies are necessary to confirm this protective effect.     

Segmentation d’images de lésions cérébrales primitives en TEP FET : influence du volume de travail

Purpose(1) Evaluate the reproducibility of segmentation methods depending on the preselection region for tumour volume determination on ¹⁸F-fluoro-ethyl-tyrosine (FET) PET. (2) Evaluate the intra and inter-operator reproducibility of the manual delineation. (3) Compare this delineation with the segmentation methods.Materials and methodsEighteen FET PET of patients with glioblastoma were analysed. Preselection regions were determined prior to any segmentation. Two physicians delineated the tumour volume manually. The tumour volume was also delineated with a threshold method (40 and 70% of SUV_max), and a random walk based method. Pearson coefficient (r) (P < 0.05 for r > 0.468) and Jaccard indices (JI) were used to compare the volumes.ResultsManual delineation was reproducible with r = 0.97 and IJ = 0.65 for intra-operator, and r = 0.76 and IJ = 0.45 for inter-operator reproducibility. The preselection regions for a given lesion were different and the segmentation varied with the preselection region: r = 0.55 JI = 0.58; r = 0.85 JI = 0.83; r = 0.70 JI = 0.39 respectively for the threshold of 40%, 70% and the random walk. The segmentation differed form de manual delineation with r = 0.37 and JI = 0.16; r = 0.54 and JI = 0.42; r = 0.43 and JI = 0.37 respectively for the threshold of 40%, 70% and the random walk.ConclusionThe reproducibility of the segmentation methods depends extensively on the preselection region. The intra-operator reproducibility of cerebral lesion delineation on FET PET is satisfactory. The inter-operator reproducibility could be improved.     

Breast cancer detection risk in screening mammography after a false-positive result

Background: False-positives are a major concern in breast cancer screening. However, false-positives have been little evaluated as a prognostic factor for cancer detection. Our aim was to evaluate the association of false-positive results with the cancer detection risk in subsequent screening participations over a 17-year period. Methods: This is a retrospective cohort study of 762,506 women aged 45–69 years, with at least two screening participations, who underwent 2,594,146 screening mammograms from 1990 to 2006. Multilevel discrete-time hazard models were used to estimate the adjusted odds ratios (OR) of breast cancer detection in subsequent screening participations in women with false-positive results. Results: False-positives involving a fine-needle aspiration cytology or a biopsy had a higher cancer detection risk than those involving additional imaging procedures alone (OR = 2.69; 95%CI: 2.28–3.16 and OR = 1.81; 95%CI: 1.70–1.94, respectively). The risk of cancer detection increased substantially if women with cytology or biopsy had a familial history of breast cancer (OR = 4.64; 95%CI: 3.23–6.66). Other factors associated with an increased cancer detection risk were age 65–69 years (OR = 1.84; 95%CI: 1.67–2.03), non-attendance at the previous screening invitation (OR = 1.26; 95%CI: 1.11–1.43), and having undergone a previous benign biopsy outside the screening program (OR = 1.24; 95%CI: 1.13–1.35). Conclusion: Women with a false-positive test have an increased risk of cancer detection in subsequent screening participations, especially those with a false-positive result involving cytology or biopsy. Understanding the factors behind this association could provide valuable information to increase the effectiveness of breast cancer screening.     

Endarterectomy patients with elevated levels of circulating IL-16 have fewer cardiovascular events during follow-up

Background and purposeIncreased interleukin 16 (IL-16) levels in carotid plaques have been associated with reduced incidence of cardiovascular (CV) events during follow-up in patients who underwent carotid endarterectomy (CEA). In the present study we aimed to determine whether high circulating levels of IL-16 also are associated with a decreased risk of CV events after CEA.MethodsPatients, who had their carotid plaques surgically removed (n = 473), were followed for a mean follow-up time of 3.1 years. Plasma levels of IL-16 the day before surgery were analyzed by proximity extension assay (PEA) and associated with the occurrence of CV events during follow-up (n = 98).ResultsHigh levels of circulating IL-16 were independently associated with a decreased risk of CV events when comparing the highest versus the lowest IL-16 tertile (hazard ratio [HR] 0.47; 95% CI 0.27–0.81; P = 0.007), as well as with CV deaths (HR 0.25; 95% CI 0.09–0.70; P = 0.008).ConclusionThese present findings indicate an association between IL-16 and less clinical complications of atherosclerosis in a population with known advanced carotid disease.     

Effect of diagnostic cone-beam computed tomography protocols on image quality,patient dose,and lesion detection

ObjectiveTo evaluate the effect of cone-beam computed tomography (CBCT) image acquisition protocols on image quality, lesion detection, delineation, and patient dose.Methods100-patients and a CTDI phantom combined with an electron density phantom were examined using four different CBCT-image acquisition protocols during image-guided transarterial chemoembolization (TACE). Protocol-1 (time: 6 s, tube rotation: 360°), protocol-2 (5 s, 300°), protocol-3 (4 s, 240°) and protocol-4 (3 s, 180°) were used. The protocols were first investigated using a phantom. The protocols that were found to be clinically appropriate in terms of image quality and radiation dose were then assessed on patients. A higher radiation dose and/or a poor image quality were inappropriate for the patient imaging. Patient dose (patient-entrance dose and dose-area product), image quality (Hounsfield Unit, noise, signal-to-noise ratio and contrast-to-noise ratio), and lesion delineation (tumor-liver contrast) were assessed and compared using appropriate statistical tests. Lesion detectability, sensitivity, and predictive values were estimated for CBCT-image data using pre-treatment patient magnetic resonance imaging.ResultsThe estimated patient dose showed no statistical significance (p > 0.05) between protocols-2 and -3; the assessed image quality between these protocols manifested insignificant difference (p > 0.05). Two other phantom protocols were not considered for patient imaging due to significantly higher dose (protocols-1) and poor image quality (protocol-4). Lesion delineation and detection were insignificant (p > 0.05) between protocols-2 and -3. Lesion sensitivities generated were 81–89% (protocol-2) and 81–85% (protocol-3) for different lesion types.ConclusionData acquisition using protocols-2 and -3 provided good image quality, lesion detection and delineation with acceptable patient dose during CBCT-imaging mainly due to similar frame numbers acquired.     

Comparaison de différentes méthodes de segmentation en TEP/TDM pour le ciblage en radiothérapie des carcinomes épidermoïdes des VADS

IntroductionTo assess interobserver variability for biological target volume (BTV) delineation and to compare the reproductibility of different semiautomatic segmentation methods in pretreatment 18-fluorodeoxyglucose positron emission tomography (PET/CT) of head and neck squamous cell carcinoma (HNSCC).Patients and methodsPatients with histologically proved HNSCC referred to the nuclear medicine service in Brest for pretreatment PET/CT were prospectively included from February 2009 to June 2010. Three nuclear medicine physicians (two specialized in oncology) delineated manually and independently BTV on each primary tumor. Four semiautomatic segmentation methods have been studied; three using a fixed threshold and one applying an adaptive threshold based on the signal-to-background ratio (Daisne). The variability between κ observers and/or methods has been assessed. The concordance between the various BTV intersections and unions has been also assessed.ResultsThirty patients (29M; 1F) were included. The primary site location was oropharynx in six patients, oral cavity in 10 patients, hypopharynx in five patients and larynx in nine patients. A statistically significant global interobserver variability (P = 0.01) was showed, but without statistically difference between the two experienced oncologists (P = 0.15). The maximal concordance of the two experienced observers with the semiautomatic methods was found for the Daisne method (CI = 61.5%; κ = 0.68), expressing a good agreement according to the Landis and Koch criteria, better than with the segmentation method using a fixed threshold with 40% of maximal signal intensity (CI = 52.1%; κ = 0.53).ConclusionOur results suggest the feasibility of achieving HNSCC BTV delineation by PET/CT using semiautomatic methods, in particular those which apply an adaptative threshold but under the supervision of an experienced operator.     

Post-chemotherapy target volumes are safe as boost volume for intact breast radiotherapy in locally advanced breast cancer

PurposeThe purpose of our study is to evaluate the challenges in identification of postoperative complexes (POC), the utility of clips in delineation of clinical target volume for boost in LABC downstaged with neoadjuvant chemotherapy (NACT) and to correlate this with patterns of recurrence.Methods and materialsLABC patients who underwent NACT followed by BCS and radiotherapy (2007–2014) were the subject of our analysis. The data on visibility and characteristics of postoperative cavity (POC), concordance of its volume with clip volume on radiation planning scan were retrieved. A 1 cm margin beyond POC was delineated as a clinical target volume (CTV). Postoperative whole breast and supraclavicular radiotherapy (50 Gy/25fractions/5wk or 42.4 Gy/16#/3 wk) followed by boost (10−16 Gy/5-8#/1–1.5wk) were delivered. Patterns of recurrence were evaluated.ResultsOut of 60 patients, 28.3% patients had stage II disease and 71.7% had stage III disease. 25% patients achieved pathological CR (complete response). The median POC volume was 30 cc and the median clip volume was 40 cc. The concordance of POC volume with clip volume was seen in 80%. Clips served as a good surrogate for POC in 80% of patients. At a median follow-up of 65 months (IQ range 32–84 months), and a lost to follow-up rate of 11.6 %, 3.3% (n = 2) patients had local recurrence (LR) and 8.3% (n = 5) had regional recurrence (LRR) in the supraclavicular region.ConclusionsDelineation of post NACT excision cavity as POC for boost radiotherapy is safe. Clips serve as a good surrogate for CTV delineation in 75% patients.     

Impact of coronal and sagittal views on lung gross tumor volume delineation

Background and purposeTo study the impact of coronal and sagittal views (CSV) on the gross tumor volume (GTV) delineation on CT and matched PET/CT scans in non-small cell lung cancer.Material and methodsGTV delineations were performed by 11 experienced radiation oncologists on CT and PET/CT in 22 patients. Two tumor groups were defined: Group I: Primary tumors surrounded by lung or visceral pleura, without venous invasion, and without large extensions to the chest wall or the mediastinum. Group II: Tumors invading the hilar region, heart, large vessels, pericardium, and the mediastinum and/or associated with atelectasis. Tumor volumes and inter-observers variations (SD) were calculated and compared according to the use of axial view only (AW), axial/coronal/sagittal views (ACSW) and ACSW/PET (ACSWP).ResultsCSV were not frequently used (57.4% out of 242 delineations on CT). For group I, ACSW didn’t improve significantly mean GTVs. SDs were small on CT and on PET (SD = 0.3 cm). For group II, ACSW had 27–46% smaller observer variation (mean SD = 0.7 cm) than AW (mean SD = 1.1 cm). The smaller observer variation of ACSW users was associated with, on average, a 40% smaller delineated volume (p = 0.038). Mean GTV of ACSWP was 21% larger than mean GTV of ACSW on CT.ConclusionsFor smaller lung tumors surrounded by healthy lung tissue the effect of multiple axis delineation is limited. However, application of coronal and sagittal windows is highly beneficial for delineation of more complex tumors, with atelectasis and/or pathological lymph nodes even if PET is used.     

HPLC with pulsed amperometric detection for sorbitol as a biomarker for diabetic neuropathy

This study describes the optimal analytical conditions for sorbitol analysis by a high-performance anion-exchange chromatography-pulsed amperometric detection method. Its clinical utility as a diagnostic tool was established by measuring sorbitol in the sciatic nerves or salivary glands of diabetes mellitus-induced mice. Sorbitol was completely separated from other monosaccharides on an anion-exchange column with 100 mM NaOH as eluent. The limit of detection (S/N = 3) and limit of quantification (S/N = 10) were 0.03 ng (3 ng/g) and 0.10 ng (10 ng/g), respectively. The linear dynamic range was 0.01–50.0 μg/g (r² = 0.9997 and 0.9989 for sciatic nerves and salivary glands, respectively), and the mean recoveries for intra- or inter-day assays were in the range of 98.5–103.9%. This method easily identified diabetic and normal groups, making it a practical procedure for the rapid screening of diabetic neuropathy.     

Novel formulation of a double threshold algorithm for the estimation of muscle activation intervals designed for variable SNR environments

The aim of this work is the development of an improved formulation of the double threshold algorithm for sEMG onset–offset detection presented by Bonato and co-workers. The original formulation, which keeps the threshold fixed, suffers from performance degradation whenever the SNR changes during the analysis. The novel approach is designed to be adaptive to SNR changes in either burst or inter-burst zones of sEMG signals recorded in static and dynamic conditions. The detection parameters (i.e. detection and false alarm probabilities) are updated on the basis of an on-line estimation of the SNR. The proposed formulation has been assessed on both simulated and real sEMG data. For constant SNR the performance of the original formulation is confirmed (for SNR > 8 dB, bias and standard deviation less than 10 and 15 ms, respectively; detection percentage higher than 95%), while the novel implementation performs better with time-varying SNR (for SNR varying in the range 10–25 dB the standard approach detection percentage decreases at 50%). Detection on signals recorded during isometric contractions at different force levels confirms the performance on simulated signals (StD = 134 ms; FP = 22%, and StD = 42 ms; FP = 2%, respectively for standard and novel implementation calculated as average on five experimental trials). The pseudo real-time detection allowed by this formulation can be profitably exploited by biofeedback applications based on myoelectric information.     

How reliable are lower limb biomechanical variables during running and cutting tasks

The purpose of this study was to compare the within- and between-days reliability of lower limb biomechanical variables collected during running and cutting tasks. Methods: 15 recreational athletes, 7 males and 8 females, took part in three testing sessions, two sessions on the same day with an hour gap and another session one week later. Kinematic and kinetic data during running and 90° side step cutting tasks gathered using a ten-camera motion analysis system (Qualisys) and a force platform (AMTI) embedded into the floor. Results: During both tasks, within-day ICC values for joint angles (ICCrun = 0.63–0.94 and ICCcut = 0.63–0.96) were higher than between days (ICCrun = 0.51–0.72 and ICCcut = 0.42–0.83). Out of five moments tested in each task, within-day ICC values (ICCrun = 0.64–0.89 and ICCcut = 0.79–0.94) were higher than between days (ICCrun = 0.58–0.91 and ICCcut = 0.83–0.92). During running task, within and between-day SEM values for joint moments ranged between (0.07–0.39 N m kg) and between (0.98°–5.14°) for joint angles. While during cutting, SEM values for moments ranged between (0.13–0.56 N m kg) and between (1.73–5.15) for joint angle measurement. The GRF data, in both tasks, were more reliable (ICCrun ⩾ 0.84 and ICCcut ⩾ 0.88) as compared to angles (ICCrun ⩾ 0.51 and ICCcut ⩾ 0.42), and moments (ICCrun ⩾ 0.58 and ICCcut ⩾ 0.79) data. These findings are relevant to those undertaking intervention studies because of the potential for large measurement variability when examining certain variables, which would then require considerable changes in these variables to show “real” effects of the interventions beyond measurement error.     

Patterns of metachronous metastases after curative treatment of colorectal cancer

BackgroundThis study aimed to provide information on timing, anatomical location, and predictors for metachronous metastases of colorectal cancer based on a large consecutive series of non-selected patients.MethodsAll patients operated on with curative intent for colorectal cancer (T_anyN_anyM₀) between 2003 and 2008 in the Dutch Eindhoven Cancer Registry were included (N = 5671). By means of active follow-up by the Cancer Registry staff within ten hospitals, data on development of metastatic disease were collected. Median follow-up was 5.0 years.ResultsOf the 5671 colorectal cancer patients, 1042 (18%) were diagnosed with metachronous metastases. Most common affected sites were the liver (60%), lungs (39%), extra-regional lymph nodes (22%), and peritoneum (19%). 86% of all metastases was diagnosed within three years and the median time to diagnosis was 17 months (interquartile range 10–29 months). Male gender (HR = 1.2, 95%CI 1.03–1.32), an advanced primary T-stage (T4 vs. T3 HR = 1.6, 95%CI 1.32–1.90) and N-stage (N1 vs. N0 HR = 2.8, 95%CI 2.42–3.30 and N2 vs. N0 HR = 4.5, 95%CI 3.72–5.42), high-grade tumour differentiation (HR = 1.4, 95%CI 1.17–1.62), and a positive (HR = 2.1, 95%CI 1.68–2.71) and unknown (HR = 1.7, 95%CI 1.34–2.22) resection margin were predictors for metachronous metastases.ConclusionsDifferent patterns of metastatic spread were observed for colon and rectal cancer patients and differences in time to diagnosis were found. Knowledge on these patterns and predictors for metachronous metastases may enhance tailor-made follow-up schemes leading to earlier detection of metastasized disease and increased curative treatment options.     

Microspheres of chitosan/carboxymethyl cashew gum (CH/CMCG): Effect of chitosan molar mass and CMCG degree of substitution on the swelling and BSA release

Chitosan/carboxymethyl cashew gum microspheres (CH/CMCG) were prepared with carboxymethyl cashew gum with two different degrees of substitution (DS) and loaded with bovine serum albumin (BSA). In water, for microspheres formed using low molar mass chitosan (LCH) sample swelling was observed for both CMCG samples and CMCG sample with higher DS showed greater swelling. Using high molar mass chitosan (HCH) sample swelling was observed only for microsphere with high DS of CMCG (DS = 0.44). At pH 7.4, the HCH sample led to a lower degree of swelling. The diffusion coefficients D_v were higher for the higher DS of CMCG in both media and the HCH sample had a lower D_v than LCH one. Faster BSA release rates were observed for beads prepared with the higher DS, whereas those prepared with DS = 0.16 took twice the time to reach similar release profiles. All microsphere systems investigated had a non-Fickian BSA release mechanism.     

Subtype analysis of Blastocystis sp. isolates from human and canine hosts in an urban community in the Philippines

Blastocystis sp. is a common gut-dwelling protist of both humans and animals. A cross-sectional survey among humans and their dogs was conducted to determine the prevalence of Blastocystis infection and to characterize the subtype (ST) distribution in an urban community in the Philippines. Fecal specimens from 1,271 humans and 145 dogs were collected and inoculated in diphasic culture medium. Prevalence of Blastocystis by culture was 13.0% (95% CI = 11.2–15.0) and 14.5% (95% CI = 9.6–21.2) for humans and dogs, respectively. A total of 168 culture isolates were genotyped using polymerase chain reaction (PCR) with seven pairs of ST-specific sequence-tagged-site (STS) primers. In humans, the ST present in this study were ST1 with 22.6% (95% CI = 17.2–29.0), ST2 with 3.1% (95% CI = 1.3–6.7), ST3 with 41.4% (95% CI = 34.9–48.6), ST4 with 14.8% (95% CI = 10.5–20.6), ST5 with 4.1% (95% CI = 2.0–8.0), and unknown ST with 13.9% (95% CI = 9.6–19.4). In dogs, the ST present in this study were ST1 with 4.3% (95% CI = 0.0–29.0), ST2 with 8.7% (95% CI = 1.3–28.0), ST3 with 17.4% (95% CI = 6.4–37.7), ST4 with 13.0% (95% CI = 3.7–33.0), ST5 with 13.0% (95% CI = 3.7–33.0), and unknown ST with 47.8% (95% CI = 29.2–67.0). This is the first study that reported Blastocystis ST4 in human and canine hosts in the Philippines.     

Syntheses and pharmacological characterization of novel thiazole derivatives as potential mGluR5 PET ligands

Four novel thiazole containing ABP688 derivatives were synthesized and evaluated for their binding affinity towards the metabotropic glutamate receptor subtype 5 (mGluR5). (E)-3-((2-(Fluoromethyl)thiazol-4-yl)ethynyl)cyclohex-2-enone O-methyl oxime (FTECMO), the ligand with the highest binding affinity (K_i = 5.5 ± 1.1 nM), was labeled with fluorine-18. [¹⁸F]-FTECMO displayed optimal lipophilicity (log D_pH7.4 = 1.6 ± 0.2) and high stability in rat and human plasma as well as sufficient stability in rat liver microsomes. In vitro autoradiography with [¹⁸F]-FTECMO revealed a heterogeneous and displaceable binding in mGluR5-rich brain regions. PET imaging with [¹⁸F]-FTECMO in Wistar rats, however, showed low brain uptake. Uptake of radioactivity into the skull was observed suggesting in vivo defluorination. Thus, although [¹⁸F]-FTECMO is an excellent ligand for the detection of mGluR5 in vitro, its in vivo characteristics are not optimal for the imaging of mGluR5 in rats in vivo.     

Between-day reliability of a hand-held dynamometer and surface electromyography recordings during isometric submaximal contractions in different shoulder positions

Functional shoulder assessments require the use of objective and reliable standardized outcome measures. Therefore, the aim of this study was to examine the between-day reliability of a hand-held dynamometer when measuring muscle strength during flexion, abduction, and internal and external rotation as well as surface electromyography (EMG) when measuring muscle activity from m. trapezius superior and deltoideus anterior. Twenty-four healthy subjects participated and performed four isometric contractions measured with a hand-held dynamometer and EMG. Both relative and absolute reliability were calculated based on the mean of the last three of the four repetitions. EMG amplitude was assessed calculating both absolute and normalized root-mean-square (RMS) values. The reliability of the hand-held dynamometer was high (LOA = 3.2–7.6% and ICC = 0.89–0.98). The absolute reliability for EMG showed similar results for absolute RMS values (LOA = 20.0–68.4%) and normalized RMS values (LOA = 42.4–66.5%). However, the results concerning the relative reliability showed higher ICC for absolute RMS values (ICC = 0.82–0.92) compared with normalized values (ICC = 0.57–0.72).The outcome measurements of this study with healthy subjects were found reliable and, therefore, have the potential to detect changes in muscle strength and muscle activity.     

Atrial flutter and atrial tachycardia detection using Bayesian approach with high resolution time–frequency spectrum from ECG recordings

Contemporary methods of atrial flutter (AFL), atrial tachycardia (AT), and atrial fibrillation (AF) monitoring, although superior to the standard 12-lead ECG and symptom-based monitoring, are unable to accurately discriminate between AF, AFL and AT. Thus, there is a need to develop accurate, automated, and comprehensive atrial arrhythmia detection algorithms using standard ECG recorders. To this end, we have developed a sensitive and real-time realizable algorithm for accurate AFL and AT detection using any standard electrocardiographic recording. Our novel method for automatic detection of atrial flutter and atrial tachycardia uses a Bayesian approach followed by a high resolution time–frequency spectrum. We find the TQ interval of the electrocardiogram (ECG) corresponding to atrial activity by using a particle filter (PF), and analyze the atrial activity with a high resolution time–frequency spectral method: variable frequency complex demodulation (VFCDM). The rationale for using a high-resolution time–frequency algorithm is that our approach tracks the time-varying fundamental frequency of atrial activity, where AT is within 2.0–4.0 Hz, AFL is within 4.0–5.3 Hz and NSR is found at frequencies less than 2.0 Hz. For classifications of AFL (n = 22), AT (n = 10) and normal sinus rhythms (NSR) (n = 29), we found that our approach resulted in accuracies of 0.89, 0.87 and 0.91, respectively; the overall accuracy was 0.88.     

A study on the association of TNF-α-308, IL-6-174, IL-10-1082 and IL-1RaVNTR gene polymorphisms with rheumatic heart disease in Pakistani patients

Inflammation is an important contributor to the pathogenesis of rheumatic heart disease (RHD), a disorder of heart valves caused by a combination of immune, genetic and environmental factors. Cytokines are important mediators of inflammatory and immune responses. The aim of this study was to investigate the role of cytokine gene polymorphisms and their potential usefulness as biomarkers in RHD patients from Pakistan. We screened 150 RHD patients and 204 ethnically matched controls for tumor necrosis factor (TNF)-α^-308G/A, interleukin (IL)-10^?1082 G/A, interleukin (IL)-6^-174 G/C and a variable number of tandem repeats (VNTRs) polymorphism of the IL-1Ra gene using polymerase chain reaction. The results showed that TNF-α^-308 A and IL-6^-174 G alleles were associated with susceptibility to RHD (p = 0.000; OR = 2.81; CI = 1.5–5.14 and p = 0.025; OR = 1.50; CI = 1.04–2.16 respectively). The TNF-α^-308 AA and GA genotypes were associated with susceptibility to RHD (p = 0.012; OR = 9.94; CI; 1.21–217.3 and p = 0.046; OR = 1.97; CI = 0.98–3.97 respectively) while the GG genotype seemed to confer resistance (p = 0.003; OR = 0.39; CI = 0.20–0.76). The GG genotype for IL-6^-174 was significantly associated with predisposition to RHD (p = 0.015; OR = 2.6; CI = 1.17–5.85). The A1 (four repeats) and A2 (two repeats) alleles at the IL-1Ra VNTR polymorphism were associated with resistance and susceptibility to RHD respectively. However, this polymorphism deviated from Hardy–Weinberg equilibrium in both patients and controls in our population. TNF-α^-308 and IL-6^-174 polymorphisms may be useful markers for the identification of individuals susceptible to RHD in Pakistan. These individuals could be provided aggressive prophylactic intervention to prevent the morbidity and mortality associated with RHD.