Identification of a New Allergen from Amaranthus retroflexus Pollen,Ama r 2

BackgroundPollinosis from Amaranthus retroflexus pollen is a common cause of respiratory allergy in Iran with a high positive rate (68.8%) among Iranian allergic patients. The aim of the present study was to evaluate the allergenicity of the A. retroflexus pollen profilin.MethodsUsing sera from twelve patients allergic to A. retroflexus pollen, IgE-binding proteins from the A. retroflexus pollen extract was identified by immunoblotting. The cDNA of A. retroflexus pollen profilin was amplified, then cloned into the pET-21b (+) vector, expressed in Escherichia coli, and finally purified by metal affinity chromatography. The IgE-binding capacity of the recombinant protein was then analyzed by the ELISA, immunoblotting, and inhibition assays, as well as by the skin prick test (SPT).ResultsImmunoblotting results indicated a 14.6 kDa protein with IgE-reactivity to 33% (4/12) among A. retroflexus pollen-allergic patients. Nucleotide sequencing of the cDNA revealed an open reading frame of 399 bp encoding for 133 amino acid residues which was belonged to the profilin family and designated as Ama r 2. A recombinant Ama r 2 (rAma r 2) was then produced in E. coli as a soluble protein which showed a strong IgEreactivity via ELISA confirmed by the SPT. Inhibition experiments revealed high IgE cross-reactivities with the profilins from other plants.ConclusionsThe profilin from the A. retroflexus pollen, Ama r 2, was firstly identified as an allergen. Moreover, rAma r 2 was produced in E. coli as a soluble immunoreactive protein with an IgE-reactivity similar to that of its natural counterpart.     

Expression of grape class IV chitinase in Spodoptera frugiperda (Sf9) insect cells

IntroductionMost of pathogenesis related (PR) proteins possess complicated structures; hence their active recombinant forms are usually produced in eukaryotic systems. In this study, we employed an insect cell line to express a recombinant form of a previously identified grape PR3 allergen categorised as class IV chitinase.MethodsGrape chitinase cDNA was amplified by RT-PCR and inserted into pFastBacHTA using restriction enzymes. The recombinant pFastBacHTA was applied for the transformation of Escherichia coli DH10Bac cells. The purified recombinant bacmid was used for transfection of Sf9 cells. Finally, the IgE-immunoreactivity of purified recombinant protein was evaluated using grape allergic patient's sera. Moreover, polyclonal anti-6His-tag and monoclonal anti-chitinase antibodies were used for further assessment of recombinant protein.ResultsSDS–PAGE analysis of the transfected Sf9 cells showed expression of a monomeric 25 kDa and a dimeric 50 kDa recombinant protein. Western blotting revealed considerable IgE reactivity of the recombinant protein with grape allergic patients’ sera. Furthermore, confirmatory assays showed specific reactivity of the recombinant protein with anti-His tag and anti-chitinase antibodies.ConclusionThis study showed that, in contrast to E. coli, insect cells are suitable hosts for the production of a soluble and IgE-reactive recombinant form of grape class IV chitinase. This recombinant allergen could be used for component resolved diagnosis of grape allergy or other immunodiagnostic purposes.     

Molecular Cloning and Expression of Cucumisin (Cuc m 1), a Subtilisin-like Protease of Cucumis melo in Escherichia coli

BackgroundOral allergy syndrome resulted from plant-derived foods is frequent among adults. Allergy to melon (cucumis melo) is one of the most frequent fruit allergies in Iran. Three different major allergens have been found in Cucumis melo that Cuc m 1 (cucumisin) has been identified as the major allergen of melon. Cucumisin is an alkaline serine protease that it is found as a 78 kDa protein in precursor form. The aim of this study was production of recombinant Cuc m 1 in Escherichia coli (E. coli) cells and characterization of its allergenicity property.MethodsProduction of recombinant Cuc m 1 was carried out by cDNA cloning technique into the pET32b(+) vector using specific primers designed based on cucumisin nucleotide sequence available in Genebank database, cucumisin encoding gene and directional cloning method. Cloned plasmid into E. coli TOP10 was transformed into E. coli BL21 and expression of the protein was induced by iptg. The recombinant protein was purified via Ni-NTA affinity chromatography using histidine tag in recombinant protein. IgE binding of this protein was assessed by IgE-immunoblotting, ELISA and inhibition ELISA.ResultsThe directional cloning was resulted in expression of a fusion Cuc m 1. Immunoblotting with sera of patients allergic to melon showed strong reactivity with purified protein band. Inhibition assays demonstrated that purified rCuc m 1 could be the same with natural form of Cuc m 1 in total extract.ConclusionsIn the present study, we have provided a functional recombinant cucumisin allergen, rCuc m 1 with 86 kDa, which may be used as a standard allergen for clinical diagnosis and study of allergy to melon.     

Decreased A20 mRNA and protein expression in peripheral blood mononuclear cells in patients with type 2 diabetes and latent autoimmune diabetes in adults

AimsA20 is a negative regulator of nuclear factor kappa B activation and the central gatekeeper in inflammation and immunity. While its role in type 1 diabetes has been widely studied, its expression level in immune cells from type 2 diabetes (T2D) and latent autoimmune diabetes in adult (LADA) patients remains unclear. This study aimed to clarify whether the expression of A20 is altered in patients with T2D or LADA.MethodsQuantitative real-time polymerase chain reaction and western blotting were utilized to determine the expression of A20 mRNA and protein respectively in peripheral blood mononuclear cells (PBMCs) from patients with T2D (n = 36) or LADA (n = 17) and sex- and age-matched healthy controls (n = 34).ResultsThe mRNA and protein expression of A20 in PBMCs from T2D and LADA patients was significantly decreased compared with healthy controls (P < 0.05). Furthermore, A20 mRNA and protein expression was significantly lower in newly diagnosed T2D patients (≤1 year since diagnosis) than in patients with a long T2D duration (>1 year since diagnosis) (P < 0.05).ConclusionsOur results suggest that decreased expression of A20 in PBMCs may be involved in the pathogenesis of diabetes, and targeting A20 may offer a potential therapeutic tool in the treatment of diabetes.     

Evaluation of serum IgE in peach-allergic patients with systemic reaction by using recombinant Pru p 7 (gibberellin-regulated protein)

Background

Lipid transfer protein (LTP) is a major fruit allergen. It has, however, recently been revealed that the systemic reaction in peach-allergic patients is related not only to LTP (Pru p 3) but also to gibberellin-regulated protein (Pru p 7). We investigated recombinant Pru p 7 (rPru p 7) for its potential use in worldwide standardization for the diagnosis of peach allergy.

Role of advanced glycation end product (AGE)-induced receptor (RAGE) expression in diabetic vascular complications

AimsVascular complications are the major causes of morbidity and mortality in diabetic subjects. Interaction of advanced glycation end products (AGEs) with their receptor (RAGE) induces signal transduction that culminates in vascular complications. Therefore, in the present study we investigated the dependence of RAGE expression on circulating AGEs and evaluated the outcome of AGE–RAGE interaction by the oxidative stress and nature of vascular complications in type 2 diabetes mellitus (T2DM) patients.MethodsRAGE expression was determined by quantitative real-time PCR and western blotting, serum AGEs were estimated by ELISA and spectrofluorometry and oxidative stress markers namely protein carbonyl (PCO), advanced oxidation protein products (AOPP) and lipid peroxidation (MDA) were assayed spectrophotometerically in 75 T2DM patients (DM without vascular complication n = 25; DM with microvascular complications n = 25; DM with macrovascular complications n = 25) and 25 healthy controls.ResultsSerum AGE level was significantly higher in diabetic patients having vascular complications as compared to T2DM without complications (p < 0.01). RAGE m-RNA expression level in PBMCs assayed by quantitative real time PCR was four times higher in diabetic subjects without vascular complications while DM patients having microvascular and macrovascular complications showed 12 fold and 8 fold higher RAGE m-RNA expression respectively compared to healthy controls. Circulating AGE level showed significant positive correlation with RAGE m-RNA expression and oxidative stress markers.ConclusionAGE-mediated exacerbation of RAGE expression may contribute to oxidative stress generation that plays a key role in pathogenesis of vascular complications in diabetes.     

Pilot Study Using Recombinant Antigens r-PROE and r-IGLL1 for the Serodiagnosis of Feather Duvet Lung

BackgroundFeather duvet lung (FDL) is an underestimated form of acute and chronic hypersensitivity pneumonitis. Serological tests for FDL need to be validated. We investigated the ability of recombinant pigeon Proproteinase E (r-PROE) and Immunoglobulin-lambda-like-polypeptide-1 (r-IGLL1) proteins to support the serological diagnosis of FDL, and propose them as a serological tool for clinicians to differentiate cases from FDL and Bird fancier's lung (BFL).MethodsSpecific IgG antibodies against r-PROE and r-IGLL1, analyzed with ELISA, were measured in patients diagnosed with FDL (n = 31), BFL (n = 15) controls exposed (n = 15) and unexposed to feathers (n = 15).ResultsThe sensitivity and specificity of the r-PROE ELISA for the serological diagnosis of FDL cases versus exposed and unexposed controls were 74.2% and 86.7% respectively, with an index threshold of 0.5 (AUC: 0.89). In addition, this serological test was effective to support the serological diagnosis of FDL and BFL cases with significantly different thresholds. The r-IGLL1 ELISA was only effective for the serological diagnosis of BFL. Also, these two serological tests were useful for the diagnosis of both chronic and acute forms.ConclusionsThe new diagnostic test for FDL using r-PROE protein should help to detect overt and hidden cases of FDL. The combination of both test will help the clinician in distinguish between the etiology of birds or feathers duvet.     

Increased plasma levels of NGAL, a marker of neutrophil activation, in patients with abdominal aortic aneurysm

ObjectiveNeutrophil gelatinase-associated lipocalin (NGAL) plasma concentrations have been associated with cardiovascular diseases. We aimed to assess the association of NGAL with abdominal aortic aneurysm (AAA).MethodsNGAL concentrations were analyzed by Western blotting in conditioned medium of polymorphonuclear neutrophils (PMNs) from AAA patients (n = 22) and controls (n = 11), and also in aortic biopsies from AAA patients and healthy controls (n = 10). Plasma NGAL concentrations were measured by ELISA in three groups of subjects from France (n = 60), Spain (n = 75) and Australia (n = 100) and associated with AAA presence and growth.ResultsPMNs isolated from AAA patients secreted significantly greater amounts of NGAL than PMNs from controls. Luminal thrombus released large amounts of NGAL compared to abluminal AAA thrombus, AAA wall and healthy aortic media. Plasma NGAL concentrations were significantly higher in patients with AAA than controls from France [115 (78–200) vs. 94 (72–114) ng/ml, p < 0.001]. NGAL plasma concentrations in AAA patients from Spain correlated with other markers of thrombus activity (plasmin–antiplasmin complexes and D-dimer). Furthermore, a positive correlation between plasma NGAL and retrospective AAA growth (rho = 0.4, p = 0.01) was observed, which remained significant after adjusting for other risk factors. Plasma NGAL was only weakly associated with prospective growth in both Spanish and Australian patients.ConclusionsNGAL is released by PMNs and by the luminal part of AAA thrombus. NGAL plasma levels were increased in AAA patients compared with healthy subjects and correlated with retrospective AAA growth. Further studies in larger subjects groups are needed to confirm the association between NGAL and AAA presence and growth.     

Plasma profiling by a protein array approach identifies IGFBP-1 as a novel biomarker of abdominal aortic aneurysm

ObjectiveCytokines are important mediators of immune-inflammatory responses implicated in abdominal aortic aneurysm (AAA) pathogenesis. Our objective was to investigate the cytokine expression profile in plasma of AAA patients.MethodsCytokine protein expression was measured in plasma of 5 large AAA patients (aortic size >50 mm) and 5 controls (aortic size <30 mm) using a 20-cytokine antibody-based protein array. IGFBP-1 plasma concentrations were analyzed by ELISA. IGFBP-1 protein levels were analyzed in AAA thrombus by immunohistochemistry and Western blot. Platelet aggregation was assessed by conventional optical aggregometry.ResultsSeveral proteins including MIP-3alpha (CCL20), Eotaxin-2 and IGFBP-1 were increased in AAA patients compared to controls. Among them, IGFBP-1 concentrations were significantly higher in large AAA patients vs control subjects. These data were validated in plasma of patients with large AAA (n = 30) compared to matched controls (n = 30) [834(469–1628) vs 497(204–893) pg/ml, p < 0.01]. Furthermore, the potential association of IGFBP-1 with AAA size was analyzed in a second independent group of subjects [large AAA (n = 59), small AAA patients (aortic size = 30–50 mm, n = 54) and controls (n = 30)]. Interestingly, IGFBP-1 levels correlated with AAA size (r = 0.4, p < 0.001), which remained significant after adjusting for traditional risk factors. IGFBP-1 was localized in the luminal part of AAA thrombus and IGFBP-1 levels were increased in AAA thrombus conditioned media compared to media layer and healthy media. Interestingly, IGFBP-1 abrogated the potentiation of ADP-induced platelet aggregation triggered by IGF-1.ConclusionsIGFBP-1 has been identified by a protein array approach as a potential novel biomarker of AAA. The biological role of IGFBP-1 in AAA pathogenesis could be related to the modulation on the effect of IGF-1 on platelet aggregation.     

Interstitial fluid contains higher in vitro IGF bioactivity than serum: A study utilizing the suction blister technique

ContextCirculating insulin-like growth factors (IGFs) are bound in complexes which affect their tissue-accessibility. Interstitial fluid is in close proximity to target cells, but the IGF-system is not well-described herein.ObjectiveTo perform a thorough comparison of the IGF-system in suction blister fluid (SBF) vs. in serum, with emphasis on bioactive IGF levels.DesignEight hour study including samples collected in the fasting state (20 h) and after a meal.SettingClinical research facility.ParticipantsSix healthy males (age 37.0 ± 8.8 years, BMI 22.5 ± 1.4 kg/m²) (mean ± SD).Main outcome measureSerum and SBF concentrations of bioactive IGF (determined in vitro by specific IGF-I receptor (IGF-IR) phosphorylation assay), immunoreactive IGF and IGF binding protein (IGFBP) levels, Western ligand blotting (WLB) of IGFBPs and IGFBP-3 Western immunoblotting (WiB).ResultsThe ability of SBF to phosphorylate the IGF-IR in vitro was 41 ± 27% higher than that of serum (P = 0.007 by repeated measures ANOVA). By contrast, immunoreactive IGF and IGFBP-concentrations were approximately 50% lower in SBF than in serum (all P ≤ 0.002). A marked difference in the composition of IGFBPs between serum and SBF was observed, including 3-fold elevated amounts of IGFBP-3 fragments in SBF (P < 0.001). For both IGF-I, IGF-II and IGFBP-2, the effect of food intake differed between serum and SBF (all P ≤ 0.03).ConclusionDespite lower concentrations, the in vitro IGF bioactivity was higher in SBF than in serum. This may relate to an increased enzymatic IGFBP-degradation and an altered IGFBP-composition in SBF, making more IGF-I and -II accessible to the IGF-IR. The impact of food intake on the IGF system differs between serum and interstitial fluid.     

Interacción cardiorrenal y evolución de la insuficiencia cardiaca. ¿Tiene un papel la proteína de unión del factor de crecimiento de tipo insulina 2?

Introduction and objectivesPreliminary results suggest that high circulating insulin-like growth factor binding protein 2 (IGFBP2) levels are associated with mortality risk in heart failure (HF) patients. As IGFBP2 levels are increased in patients with chronic kidney disease (CKD), which is associated with a higher mortality risk in HF patients, we examined whether IGFBP2 is associated with CKD in HF patients, and whether CKD modifies the prognostic value of this protein in HF patients.MethodsHF patients (n = 686, mean age 66.6 years, 32.7% women) were enrolled and followed up for a median of 3.5 (min-max range: 0.1-6) years. Patients were classified as having CKD with decreased estimated glomerular filtration rate (eGFR < 60 mL/min/1.73 m²) or as having CKD with nondecreased eGFR (≥ 60 mL/min/1.73 m²). Serum IGFBP2 was detected by ELISA.ResultsIGFBP2 was increased (P < .001) in CKD patients with decreased eGFR (n = 290, 42.3%) compared with patients with nondecreased eGFR. IGFBP2 was directly associated with NT-proBNP (P < .001) and inversely associated with eGFR (P < .001), with both associations being independent of confounding factors. IGFBP2 was directly and independently associated with cardiovascular and all-cause death (P < .001) in the whole group of patients, but showed a stronger association with cardiovascular death in CKD patients with decreased eGFR (P for interaction < .05), improving risk prediction in these patients over clinically relevant risk factors.ConclusionsSerum IGFBP2 is associated with impaired renal function and prognosticates cardiovascular death in patients with HF and CKD with decreased eGFR. Thus, there is an effect modification of CKD on circulating IGFBP2 and on its association with cardiovascular mortality in HF patients.     

A comparative analysis of novel cardiovascular biomarkers in patients with chronic heart failure

BackgroundHeart failure (HF) with reduced ejection fraction remains a major therapeutic challenge. The aim of this study was to investigate the role of novel cardiovascular biomarkers, i.e. soluble suppression of tumorigenicity (sST2), growth-differentiation factor-15 (GDF-15), soluble urokinase plasminogen activator receptor (suPAR) and heart-type fatty acid binding protein (H-FABP) in patients with ischaemic (ICM) or dilative cardiomyopathy (DCM).Materials and methodsA total of 200 patients were enrolled in this study: 65 were diagnosed with DCM and 59 patients suffering from ICM were included. 76 patients without coronary artery disease or signs of heart failure were included as controls. Plasma samples of all patients were analyzed by use of ELISA.ResultsLevels of sST2, suPAR and H-FABP were significantly higher in ICM and DCM patients compared to the control group (p < 0.0001). However, there were no significant differences between ICM and DCM in biomarker levels. Ejection fraction correlated inversely with cardiac biomarkers (sST2 p < 0.0001, GDF-15 p = 0.0394, suPAR p = 0.0029, H-FABP p < 0.0001). Similarly, CRP levels also showed a positive correlation with cardiac biomarkers. Renal insufficiency (p < 0.0001) and diabetes (sST2 p = 0.0021, GDF-15 p = 0.0055, suPAR p = 0.0339, H-FABP p = 0.0010) were significantly associated with a rise in cardiac biomarkers.ConclusionNovel cardiovascular biomarkers such as ST2, GDF-15, uPAR and H-FABP could offer a great potential for more precise diagnostic in ICM and DCM patients. H-FABP was the most promising marker in our study, followed by sST2, uPAR and GDF-15. Additional prospective studies will be necessary to further evaluate the potential clinical benefits in routine treatment of HF.     

Endothelial function parameters in patients with unstable angina and infection with Helicobacter pylori and Chlamydia pneumoniae

BackgroundEndothelial dysfunction may be a factor linking infection with atherosclerosis. The aim of our study was to assess the relationship between seropositivity to Helicobacter pylori (Hp) and/or to Chlamydia pneumoniae (Cp) and some endothelial function parameters in patients with unstable angina.MethodsIn 31 patients with unstable angina, we determined the serum concentration of the von Willebrand factor (vWF), thrombomodulin, tissue plasminogen activator antigen, and tissue plasminogen activator inhibitor type 1 antigen, the concentration of IgG antibodies to Hp and Cp (all by ELISA), and the level of C-reactive protein. The Western blot test was performed for all patients seropositive to Hp. It allowed us to identify 15 different antigen proteins of Hp.ResultsSixty-one percent of the patients were seropositive to both Hp and Cp, and 35% were seropositive to Hp only. We did not find significant differences in serum concentrations of endothelial function parameters and CRP between the two groups of patients. The patients seropositive to both Hp and Cp had a significantly higher serum concentration of vWF when Hp did not contain the 95 kDa protein (p = 0.01) and a significantly higher serum concentration of PAI-1:Ag when Hp did not contain the 57 kDa protein (p = 0.002) and the 66 kDa protein (p = 0.02).ConclusionThe results show that the antigenic profile of bacteria may play a more significant role in coronary artery disease than seropositivity.     

Vitamin D levels in patients with Behçet’s disease: Significance and impact on disease measures

Aim of the workThis study aimed to investigate serum levels of vitamin D in patients with Behçet’s disease (BD) and to evaluate their relationship to disease activity as well as different disease measures.Patients and methodsForty-two patients with BD were enrolled into this study. These patients were subjected to detailed history taking, thorough clinical examination including assessment of disease activity according to Behçet’s Disease Current Activity Form (BDCAF) score and performed laboratory investigations including erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), serum calcium, serum phosphorus and serum alkaline phosphatase. Serum 25-hydroxyvitamin D (vitamin D) levels were determined using Enzyme-Linked-Immunosorbent Assay (ELISA). A control group of 41 age and sex matched healthy controls was also included.ResultsThe mean level of 25-hydroxyvitamin D (30.65 ± 12.87 ng/ml) was significantly decreased in BD patients compared to the controls (37.98 ± 15.76 ng/ml) (p = 0.02). Significant negative correlations of serum vitamin D levels with patients’ ages (p = 0.03), ESR (p < 0.001), CRP (p < 0.001) and BDCAF (p = 0.003) were found; whereas, there was no significant correlation with disease duration (p = 0.6). In multivariate regression analysis age (p = 0.02), colchicine therapy (0.008), ESR (0.02) and CRP (0.03) were found to be the independent effectors on vitamin D serum levels.ConclusionSerum levels of vitamin D were significantly lower in BD patients compared to controls. Associations were found between vitamin D levels and age, BDCAF as well as ESR and CRP in BD patients. Low vitamin D may predispose BD patients to active disease, especially in older subjects.     

Change in tear protein profile in diabetic retinopathy with duration of diabetes

AimsTo study change in tear protein profile with duration of diabetes and severity of diabetic retinopathy (DR) in type 2 diabetes patients.Materials and methodsTear protein profile was ascertained by SDS PAGE method in 30 patients with DR (group A) and 37 patients without DR (group B).ResultsSix distinct bands of proteins were identified; these proteins are as follows: 91 kDa (P1), 66 kDa (P2), 60 kDa (P3), 30 kDa (P4), 18.4 kDa (P5) and 14.4 kDa (P6). Prevalence of P3 was significant (p = 0.036) in group A, especially in cases with diabetes ≤8 years compared with diabetes >8 years (p = 0.0107). In group B, P2 was significantly prevalent (p < 0.0013) in cases with diabetes ≤8 years compared to diabetes >8 years. Considering the changes in terms of duration of diabetes in general, patients with diabetes of ≤8 years, P3 was significantly prevalent in group A compared to group B (p = 0.004); and when the duration of diabetes is >8 years, P2 was found significantly more in group A compared to group B (p = 0.01). No significant difference in P3 (p = 0.025), P4 (p = 0.2877), P5 (p = 0.4801), P6 (p = 0.0985) was observed in mild to moderate NPDR group compared to severe NPDR to PDR group. P1 and P2 were present only in severe NPDR and PDR.ConclusionVariable protein expression was observed with duration of diabetes and severity of diabetic retinopathy.     

Orexin A associates with inflammation by interacting with OX1R/OX2R receptor and activating prepro-Orexin in cancer tissues of gastric cancer patients

ObjectiveGastric cancer (GC) has been become the second leading cause for cancer-associated death. This study aimed to investigate Orexin A levels and associated receptors in tumor tissues of GC patients.Patients and methodsForty-six consecutive gastric cancer patients (GC, n = 46) and 13 chronic atrophic gastritis patients (CAG, n = 13) were recruited. Meanwhile, 18 health individuals visiting Medical Examination Department were involved as control (N group, n = 18). ELISA was used to examine Orexin A concentration. Immunohistochemistry assay was used to examine OX1R and OX2R. HE staining was applied to evaluate inflammation. qRT-PCR was employed to detect OX1R, OX2R, prepro-Orexin mRNAs. Serum Helicobacter pylori (H. pylori) infection was measured.ResultsOrexin A expression in GC patients was significantly up-regulated compared to N group and CAG group (p < 0.05). Orexin A expression was increased in CAG group compared to N group (p < 0.05). Gastric cancer tissues exhibited significantly obvious inflammation compared to N group and CAG group (p < 0.05). OX1R and OX2R expressions were significantly down-regulated in GC group compared to N group and CAG group (p < 0.05). OX1R and OX2R were lower significantly in GC group compared to CAG group (p < 0.05). Prepro-Orexin was significantly depleted in tumor tissues of GC group compared to N group and CAG group (p < 0.05). Orexin A expression was un-associated with gender, age and differential grades (p > 0.05). CAG and GC patients demonstrated higher H. pylori infection rates.ConclusionOrexin A was associated with inflammation by interacting with OX1R/OX2R receptor and activating prepro-Orexin in tumor tissues of gastric cancer patients.     

CXC ligand 13 in rheumatoid arthritis and its relation to secondary Sjögren’s syndrome

Aim of the workThe aim of the present study was to measure the level of the chemokine CXC ligand 13 protein (CXCL13) in the plasma and unstimulated saliva of rheumatoid arthritis (RA) patients in order to find out its role in the disease activity and its relation to secondary Sjögren’s syndrome (sSS).Patients and methodsThe study was conducted on thirty rheumatoid arthritis patients attending the Outpatient Clinic of Rheumatology and Rehabilitation department of Ain shams University Hospitals. The patients’ group had been classified into group (1) which included fifteen RA patients associated with sSS diagnosed according to the American–European Consensus Group Classification Criteria and group (2) which included fifteen RA patients not associated with sSS. Ten healthy subjects were included as a control group. Patients were subjected to full history taking, clinical examination, and laboratory detection of CXCL13 level in the plasma and saliva of patients as well as the control groups using ELISA technique. Assessment of disease activity in RA patients was done using the disease activity score (DAS28).ResultsPlasma levels of CXCL13 were significantly higher in RA patients than control group (p < 0.001). Plasma levels of CXCL13 were significantly correlated with the RA disease activity (r = 0.677, p < 0.001) and disease duration (r = 0.406, p < 0.05), while the salivary levels were higher in those with sSS and correlated with sSS disease duration (r = 0.536, p < 0.05). A highly significant correlation was found between salivary CXCL13 and severity of sSS (r = 0.816, p < 0.001). Salivary levels of CXCL13 above 110 pg/ml may diagnose sSS with sensitivity 80% and specificity 84%.ConclusionThe results of this preliminary study point out the importance of CXCL13 as a marker for RA disease activity, its role in diagnosing sSS, and estimation of sSS severity.     

Homocysteine and endothelial markers are increased in patients with chronic liver diseases

BackgroundHyperhomocysteinaemia is a disorder of methionine metabolism, in which a liver plays a role. It may be frequently due to nutritional deficiencies, particularly low folate status. The aim of the study was to evaluate the serum concentration of homocysteine (Hcy) in patients with chronic liver diseases (CLD), and to assess the relation between Hcy, folate levels, and endothelial markers.MethodsSeventy-one patients with CLD and 51 healthy subjects of similar sex and age were investigated. There were 19 patients with steatosis and 52 patients with fibrosis/cirrhosis, classified by the Child–Pugh score as groups A, B and C. Fasting serum Hcy and folate levels were measured by the IMx diagnostic system (ABBOTT, USA). Plasma thrombomodulin (TM) and von Willebrand factor (vWF) as markers of endothelial dysfunction/damage were determined by ELISA methods.ResultsA significant increase of Hcy in all groups of patients with CLD was found: steatosis (P = 0.0036), fibrosis/cirrhosis — groups A, B and C (P = 0.0067, P < 0.0001, P = 0.0005, respectively). No significant changes of serum folate in CLD patients were observed, but there was an inverse correlation between Hcy and folate concentrations (r² = 0.1076, P = 0.0003). A significant increase of endothelial markers in CLD patients was found: TM in steatosis (P = 0.029), fibrosis/cirrhosis — group A (P = 0.0010), groups B and C (P < 0.0001, respectively), vWF in fibrosis/cirrhosis — group A (P = 0.0003), groups B and C (P < 0.0001, respectively). No significant correlation between serum Hcy and endothelial markers was observed.ConclusionHyperhomocysteinaemia and abnormalities of endothelial function are demonstrated in CLD patients. The impairment of liver metabolism and local changes in vessel integrity are supposed to play a main role.     

Antibacterial activity of Citrus hystrix toward Salmonella spp. infection

IntroductionCitrus hystrix is widely used by Indonesians as a traditional medicine for gastrointestinal diseases, including Salmonella spp. infection. We investigated the antibacterial activity of the ethanolic peel extract of C. hystrix against Salmonella typhimurium.MethodsThe antibacterial activity was evaluated both in vitro and in vivo. The minimum inhibitory concentration (MIC) of the extract was determined at a concentration of 0.625% by agar dilution assay. Later, the in vivo antibacterial activity was examined by the administration of 16 mg of the extract daily for three consecutive days in a mouse model infected with S. typhimurium.ResultsThe bacterial loads of S. typhimurium in the ileum, liver, and spleen decreased after 24 h of administration of the extract (p = 0.00008, p = 0.00084, and p = 0.00003, respectively).ConclusionThe ethanolic peel extract of C. hystrix shows antibacterial activity against S. typhimurium, indicating the potential of C. hystrix as an effective treatment for Salmonella spp. infection.     

Expression of soluble sCD163 in serum of psoriatic patients is modulated by Goeckerman therapy

BackgroundCD163 is the monocyte/macrophage receptor for haptoglobin–haemoglobin complexes. The aim of this study was to assess the kinetics in the expression of CD163 on monocytes and the concentration of soluble sCD163 in serum of psoriatic patients in order to examine the effect of Goeckerman therapy.MethodssCD163 was measured in 71 patients before and after therapy, and in 57 healthy donors. A subgroup of 40 patients and 25 controls was used to assess the expression of membrane CD163. sCD163 was evaluated by ELISA. Flow cytometry method was used to determine the expression of membrane CD163 on monocytes, expressed as mean fluorescence index (MFI).ResultsBefore therapy, the serum level of sCD163 was significantly higher in our patients than in controls (P = 0.0154). However, we observed a profound decrease in sCD163 in our patients after therapy (P = 0.0037). Similar to sCD163, pre-treatment expression of CD163 on monocytes was significantly more enhanced in patients than that in controls (P = 0.0078). There was a trend towards down-regulation of the expression after therapy, nonetheless, the change was not statistically significant compared to the values before therapy (P = 0.8666). This was also confirmed by comparison with controls which displayed lower expression of CD163 than patients after therapy (P = 0.0019). The disease activity, expressed as PASI score, was significantly decreased in our patients by GT (P = 0.0001).ConclusionsWhile sCD163 level in psoriatic patients was diminished after GT therapy, CD163 expression on monocytes was altered only to a minor extent.