A cross-sectional study on activity impairment in primary Sjogren's syndrome.

OBJECTIVE: The aim of this cross-sectional study was to show relations between activity impairment and salivary gland involvement for patient empowerment in primary Sjogren's syndrome (pSS). METHODS: In the study, 86 patients with pSS were included. The data were collected through clinical examinations and a questionnaire regarding Work Productivity and Activity Impairment (WPAI), EULAR Sjogren's syndrome patient-reported index (ESSPRI) and Oral Health Impact Profile-14 (OHIP-14). Relations were analysed by using mediation and moderation analyses. In simple mediation analysis, an independent variable (X) influences outcome variable (Y) through a mediator variable (M) whereas a moderator variable (W) affects the direction of the relationship between the dependent (Y) and independent variables (X). RESULTS: Increases in ESSPRI-Dryness score (X) (p = 0.0189) and OHIP-14 score (M) (p = 0.0004) were associated with the poor WPAI activity impairment score (Y) in the first mediation analysis. The WPAI activity impairment score was mediated by the elevated ESSPRI-Fatigue score (X) (p = 0.03641) and low U-SFR (M) (p = 0.0000) in the second mediation analysis. In addition, ESSPRI-Pain score (W) was the significant moderator for WPAI activity impairment (Y) in patients without hyposalivation in the moderation analysis (p = 0.0010). CONCLUSION: WPAI activity impairment was affected by both ESSPRI-Dryness with OHRQoL and ESSPRI-Fatigue with SFR in glandular involvement.

Evaluation of fecal calprotectin as a marker of gastrointestinal inflammation in rosacea: A case-control study.

A higher incidence of gastrointestinal diseases has been well established in patients with rosacea. However, no screening tool has been introduced for gastrointestinal disease development in rosacea. Fecal calprotectin (FC) is a calcium-binding protein, mainly derived from polymorpho-nuclear cells, such as neutrophils. It has been established as a marker of gastrointestinal inflammation. The aim of the present study was to evaluate FC levels in patients with rosacea without any gastrointestinal diseases. A prospective, case-control study was planned to investigate the relationship between rosacea and gastrointestinal involvement by evaluating FC levels and the Gastrointestinal Symptom Rating Scale (GSRS). A total of 47 patients with rosacea and 39 healthy control subjects were included in the study. The FC levels were statistically significantly higher in rosacea group than in the control group (65.96 ± 58.86 ng/mL vs 31.99 ± 20.12 ng/mL, P = .026, respectively). A statistically significant difference was also observed in GSRS values between the patient and the control groups (30.26 ± 12.48 vs 22.62 ± 7.64, P = .001, respectively). A positive correlation was noted between FC levels and the values of GSRS in the study group (r: 0.354; P = .001) and in the rosacea group (r = 0.392, P = .006). The measurement of FC may be useful in the early detection of gastrointestinal system diseases that may accompany rosacea and may provide a pathway to develop treatment strategies targeting both skin and intestinal mucosa.

Appraising diagnostic performance of ELF test by pathological staging and digital quantification of liver fibrosis.

INTRODUCTION AND OBJECTIVES: A crucial issue when appraising the performance of non-invasive markers is the limitations of the reference standard they are compared to. Digital image analysis (DIA) was suggested as a reproducible approach expressing fibrosis numerically as a proportionate area (PA) (%). We aimed to evaluate ELF test with direct reference to PA (%), thereby explore the improvement in accuracy to discriminate significant fibrosis which may actually have been underestimated by categorical pathological staging. MATERIALS AND METHODS: PA (%) data were obtained by DIA of trichrome-stained liver biopsies of 52 chronic hepatitis patients. Paired serum samples of patients and additional 36 controls were performed to measure ELF test. Diagnostic performance characteristics of ELF test was derived in predicting significant fibrosis in the patient cohort, and also, in distinguishing healthy controls from patients with significant fibrosis. RESULTS: We found an AUROC value of 0.73 for ELF to predict significant fibrosis as assessed by DIA and a lower AUROC value of 0.66 when assessed by conventional pathology. Importantly, ELF test provided considerably high diagnostic accuracy to discriminate healthy controls from patients with significant fibrosis defined by Ishak F≥2 and TPA≥5% (AUROCs 0.93 and 0.94, respectively) with optimal ELF cut-off point of 8.4 for both. CONCLUSIONS: Digital quantification could represent a better reference standard than conventional pathology allowing a better discriminatory capability for ELF test. ELF test provided high diagnostic accuracy to discriminate healthy controls from patients with significant fibrosis suggesting a role as a screening strategy in the community setting.

Evaluation of salivary gland ultrasonography in primary Sjögren's syndrome: does it reflect clinical activity and outcome of the disease?

OBJECTIVES: To evaluate associations between salivary gland ultrasonography (SGUS) and clinical characteristics, disease activity and outcome in patients with primary Sjögren's syndrome (pSS). METHODS: The parotid and submandibular salivary glands were examined by ultrasonography using two different scoring systems proposed by Hocevar et al. and Milic et al. on 85 pSS patients. Patients with inhomogeneity/hypoechoic areas with scores ≥2 in parotid and submandibular glands were classified as severe parotid or severe submandibular involvements, respectively. Disease activity and patient-reported severity were evaluated using the European League Against Rheumatism Sjögren's Disease Activity Index (ESSDAI) and the European League Against Rheumatism Sjögren's Patient Reported Index (ESSPRI). Salivary gland functional capacity was investigated by unstimulated whole saliva flow rate (U-WSFR). RESULTS: Of the activity scores, ESSPRI dryness component was higher in pSS patients who had scores above the cut-off values for Hocevar (6.1±2.3 vs. 4.9±2.6, p=0.026). The patients with any type of systemic involvement more frequently showed higher SGUS scores, according to both Hocevar (72.4 vs. 44.6%, p=0.013) and Milic (75.9 vs. 51.8%, p=0.026). These patients also showed a higher percentage of severe parotid/submandibular changes on US imaging (65.5 vs. 33.9%, p=0.005 and 75.9 vs. 51.8%, p=0.026 respectively). Higher SGUS scores according to cut-off values of both scoring systems and severe parotid/submandibular involvements were associated with both anti-Ro or double anti-Ro/La autoantibodies and inversely associated with U-WSFR. CONCLUSIONS: SGUS may be a useful imaging modality for the selection of patients with more severe disease status or who may require a tight follow-up schedule.

Sequential Measurements of Pentraxin 3 Serum Levels in Patients with Ventilator-Associated Pneumonia: A Nested Case-Control Study.

PURPOSE: The main purpose of this study was to investigate the dynamics of pentraxin 3 (PTX3) compared with procalcitonin (PCT) and C-reactive protein (CRP) in patients with suspicion of ventilator-associated pneumonia (VAP). MATERIALS AND METHODS: We designed a nested case-control study. This study was performed in the Surgical Intensive Care Unit of a tertiary care academic university and teaching hospital. Ninety-one adults who were mechanically ventilated for >48 hours were enrolled in the study. VAP diagnosis was established among 28 patients following the 2005 ATS/IDSA guidelines. RESULTS: The median PTX3 plasma level was 2.66 ng/mL in VAP adults compared to 0.25 ng/mL in non-VAP adults (p < 0.05). Procalcitonin and CRP levels did not significantly differ. Pentraxin 3, with a 2.56 ng/mL breakpoint, had 85% sensitivity, 86% specificity, 75% positive predictive value, and 92.9% negative predictive value for VAP diagnosis (AUC = 0.78). CONCLUSIONS: With the suspicion of VAP, a pentraxin 3 plasma breakpoint of 2.56 ng/mL could contribute to the decision of whether to start antibiotics.

Th17-Inducing Conditions Lead to in vitro Activation of Both Th17 and Th1 Responses in Behcet's Disease.

OBJECTIVES: Interleukin-17 (IL-17) has been associated with the pathogenesis of various autoimmune/inflammatory diseases. The aim of this study was to investigate the expression of Th17-related immunity in an innate immunity-dominated vasculitis, namely Behcet's disease (BD). METHODS: Peripheral blood mononuclear cells from 37 patients (age: 38.5 ± 9.8 years) with BD, and 25 healthy controls (HC) (age: 39.1 ± 9.3 years), were cultured in Th17-inducing conditions (IL-6, Phytohemagglutinin (PHA), IL-1ß, and IL-23) for 6 days. Cultured cells were stained with CD4, CD8, CD3, TCR gamma/delta, CD19, interferon-γ (IFN-γ), and IL-17 antibodies to determine the intracellular cytokine secretion by flow cytometry. RESULTS: IL-17 expression by CD8+ and γδ+ T cells was higher in BD compared to HC (p = 0.004, p = 0.003, respectively). No differences were observed between the groups in the IL-17 production by B cells. Under Th17-inducing conditions, production of IFN-γ by CD4+, CD8+, and γδ+ T cells was also higher in BD compared to HC (p < 0.05 in all). CONCLUSION: Our results suggest that under Th17-stimulating conditions, T cells express both IL-17 and IFN-γ in BD. More prominent IL-17 and IFN-γ production by all lymphocyte subsets in BD might be associated with the increased innate responses, early tissue neutrophil infiltrations and late adaptive immunity in BD.

Sequencing of 16S rRNA reveals a distinct salivary microbiome signature in Behçet's disease.

Behçet's disease (BD) is characterized by recurrent oro-genital ulcers, mucocutaneous lesions, and serious organ involvement. We investigated the salivary microbiome in BD using high-throughput sequencing of the 16S rRNA V4 region. Stimulated saliva samples were collected from 31 BD patients and 15 healthy controls, and in 9 BD patients, a second saliva sample was collected following dental and periodontal treatment. Sequence analysis identified a total of 908 operational taxonomic units (OTUs) present across all samples. Patients had a microbial community structure that is significantly less diverse than healthy controls. The most overabundant species in BD was Haemophilus parainfluenzae, while the most depleted included Alloprevotella rava and species in the genus Leptotrichia. Periodontal treatment improved oral health indices in BD but had no short-term effect on bacterial community structure. Neither the BD-associated genetic risk locus within the HLA-B/MICA region nor being on immunosuppressive medications explained the differences between patients and controls.

Epigenome-wide scan identifies a treatment-responsive pattern of altered DNA methylation among cytoskeletal remodeling genes in monocytes and CD4+ T cells from patients with Behçet's disease.

OBJECTIVE: The pathogenesis of Behçet's disease (BD), an inflammatory disease with multisystem involvement, remains poorly understood. This study was undertaken to investigate whether there are DNA methylation abnormalities in BD that might contribute to the pathogenesis of the disease. METHODS: We examined genome-wide DNA methylation patterns in monocytes and CD4+ T cells from 16 male patients with untreated BD and age, sex, and ethnicity-matched healthy controls. Additional samples were collected from 12 of the same BD patients after treatment and achievement of disease remission. Genome-wide DNA methylation patterns were assessed using the Infinium HumanMethylation450 BeadChip array, which includes >485,000 individual methylation sites across the genome. RESULTS: We identified 383 CpG sites in monocytes, and 125 in CD4+ T cells, that were differentially methylated between BD patients and controls. Bioinformatic analysis revealed a pattern of aberrant DNA methylation among genes that regulate cytoskeletal dynamics, suggesting that aberrant DNA methylation of multiple classes of structural and regulatory proteins of the cytoskeleton might contribute to the pathogenesis of BD. Further, treatment of BD modified the differences in DNA methylation observed in patients compared to controls; indeed, among CpG sites that were differentially methylated after disease remission versus before treatment, there was widespread reversal of the direction of aberrant DNA methylation observed between the patient and control groups. CONCLUSION: The results of this epigenome-wide study-the first such study in BD-provide strong evidence that epigenetic modification of cytoskeletal dynamics underlies the pathogenesis of BD and its response to treatment.

Pro-inflammatory cytokine and caspase-1 responses to pattern recognition receptor activation of neutrophils and dendritic cells in Behcet's disease.

OBJECTIVE: Activated innate immunity is implicated in the pathogenesis of Behcet's disease (BD). To clarify the mechanisms of innate immune responses, we investigated inflammasome activation in dendritic cells (DCs) and neutrophils, following stimulation with two different pattern recognition receptors (PRRs) RIG-1-like (RLR) and NOD-like (NLR) in patients with BD. METHODS: Sixteen active BD patients with mucocutaneous lesions and 17 healthy controls (HCs) were included in this study. DCs were generated from monocytes. DCs and isolated neutrophils were activated by RLR and NLR ligands. Caspase-1 activation and expression of p38 and RIP2 were determined by flow cytometry. Levels of IL-1ß, IL-6, TNF-α, IFN-α and IL-18 in culture supernatants were measured by ELISA. RESULTS: Activation of caspase-1 following intracellular PRR stimulation was found to be of similar levels in DCs and neutrophils of BD patients compared with HCs. However, activation of DCs from BD patients to NOD2 stimulus measured by the expression of RIP2 and p38 as well as IL-18 levels was found to be slightly defective (P < 0.05). In neutrophil cultures, IL-6 levels were lower in response to all stimuli in patients with BD compared with HCs (P < 0.01). CONCLUSION: Inflammasome formation following stimulation with NOD1/NOD2 and RIG measured by caspase-1 activation, cytokine levels and expression of RIP2 and p38 seems to be functionally normal in DCs and neutrophils of BD patients, although slightly defective responses in some pathways and cytokine levels were observed. These results may suggest that caspase-1-independent pathways such as toll-like receptors may be more prominent in BD pathogenesis.

Salivary levels of HNP 1-3 are related to oral ulcer activity in Behçet's disease.

BACKGROUND: Saliva contains antimicrobial peptides derived from oral epithelium as well as neutrophils in the innate immune response. The aim of this study was to examine the association between salivary human neutrophil peptide (HNP) 1-3 levels originating from neutrophils and oral ulcers in patients with Behçet's disease (BD). METHODS: Ninety-five patients with BD (F/M: 39/56; mean age: 38.7 ± 11.9 years) and 53 healthy controls (HC; F/M: 23/30; mean age: 35.2 ± 10.1 years) were included in the study. The disease control group (F/M: 20/33; mean age: 33.7 ± 10.7 years) was comprised of patients with oral infection regarding endodontic infection (n = 32) and pericoronitis (n = 21). Salivary HNP 1-3 levels of groups were measured in unstimulated samples by ELISA (Hycult, the Netherlands). RESULTS: A statistically significant increase was found in salivary HNP 1-3 levels of patients with BD (2268.28 ± 1216.38 µg/ml) compared with HC (1836.49 ± 857.76 µg/ml), patients with endodontic infection (849.9 ± 376.1 µg/ml), and patients with pericoronitis (824.3 ± 284.02 µg/ml; P = 0.024, 0.000 and 0.000, respectively). The ratio of active oral ulcer (100%, n = 14) was higher in low HNP 1-3 levels (≤ 1000 µg/ml) than the others (66.7%, n = 54) in active patients with BD (P = 0.008). Moreover, salivary HNP 1-3 levels were significantly lower in patients with endodontic infection and patients with pericoronitis compared with those in the HC group and patients with BD (P = 0.000). CONCLUSION: A decrease in salivary HNP 1-3 levels might be a biological factor for predisposition to oral ulcers in patients with BD and oral infection in healthy patients.

Salivary levels of antimicrobial peptides Hnp 1-3, Ll-37 and S100 in Behcet's disease.

BACKGROUND: Oral ulcer is the cardinal clinical sign and increased neutrophilic activity is a part of the pathogenesis in Behcet's disease (BD). Saliva, as a part of the innate immune response, contains antimicrobial peptides (AMPs) that are derived from both oral epithelial cells and neutrophils. The aim of this study was to investigate the associations between salivary levels of AMPs HNP 1-3, LL-37 and S100 and disease course in patients with Behcet's disease (BD). METHODS: Fifty-three patients with BD and 44 healthy controls (HC) were included in the study. Disease severity score reflecting organ involvement was calculated. Salivary HNP 1-3, LL-37 and S100 levels were measured in unstimulated saliva samples by ELISA. RESULTS: Salivary HNP 1-3 and S100 levels in BD patients (2715.2 ± 1333.4 µg/ml and 430.6 ± 203.9 ng/ml) were significantly higher compared to HC (1780.6 ± 933.2 µg/ml and 365.3 ± 84.7 ng/ml) (p = 0.000 and p = 0.004, respectively). Although LL-37 levels were also higher in BD than HC (190.9 ± 189.1 vs 143.1 ± 128.9 ng/ml), no significant difference was observed (p = 0.53). Salivary HNP 1-3 and LL-37 levels were associated with the severity of BD (mild disease: 1975.1 ± 1174.2 µg/ml and 115.9 ± 109.4 ng/ml vs severe disease: 2955.7 ± 1305.6 µg/ml and 215.3 ± 203.8 ng/ml, p=0.020 and p=0.031, respectively). Salivary LL-37 levels also correlated with the number of monthly oral ulcers (r = 0.5 p = 0.000). CONCLUSION: An increase in salivary HNP 1-3 and S100 levels might be associated with enhanced local and systemic innate responses in BD.

Association of polymorphisms of NOD2, TLR4 and CD14 genes with susceptibility to gastric mucosa-associated lymphoid tissue lymphoma.

BACKGROUND: The clinical course of Helicobacter pylori infection is highly variable and is influenced by both microbial and host factors, including the genetic composition of the infecting strains and variations in the host immune responses. A genetic risk profile for gastric cancer has been identified, but genetic susceptibility to develop gastric mucosa-associated lymphoid tissue (MALT) lymphoma is unclear. The aim of this study was to evaluate the relationship between NOD2, TLR4 and CD14 genetic polymorphisms and the development of gastric MALT-lymphoma. MATERIALS AND METHODS: Fifty-six patients with primary gastric MALT lymphoma and 51 patients with H. pylori infection were enrolled in this study. The polymorphisms were detected by the PCR-restriction fragment length polymorphism (RFLP) method of allele-specific PCR. RESULTS: No polymorphisms in the NOD2 and TLR4 genes were found to be associated with the development of gastric MALT lymphoma. Carriers of the CD14 gene -159T allele had a marginally increased risk of developing gastric MALT lymphoma than the controls (p=0.042). CONCLUSION: The -159C/T genetic polymorphism of the CD14 gene may be implicated in the development of gastric MALT lymphoma.