Whole peripheral blood miR-146a and miR-155 expression levels in Systemic lupus erythematosus patients.

Objective: To evaluate the diagnostic value of peripheral blood microribonucleic acid (miRNA, miR)-146a and miR-155 expression in systemic lupus erythematosus (SLE). Methods: Expression levels of miR-155 and miR-146a in whole peripheral blood samples from 40 SLE patients and 32 healthy controls (HCs) were determined by quantitative reverse transcription-polymerase chain reaction qRT-PCR (SYBR Green technology) and 2-∆∆Ct method was used for analysis. SPSS v20 was used for receiver operating characteristic (ROC) curve and Spearman correlation analysis. Results: Whole peripheral blood expression levels of miR-146a and miR-155 were overexpressed in 62.5% and 50%, respectively, of the SLE patients compared to HCs. The ROC curve analysis showed that the expression levels of miR-146a could discriminate SLE patients from HCs with area under the curve (AUC)=0.711 (95% CI: 0.585÷0.837, p=0.002, with 82.5% sensitivity and 56.2% specificity. The diagnostic accuracy of miR-155 was lower with AUC=0.691 (95% CI: 0.566÷0.817, p=0.005, with 77.5% sensitivity and 50.0% specificity. The diagnostic accuracy did improve when combination of the studied miRNAs was used in multimarker ROC curve analysis (AUC=0.716, 95% CI: 0.590÷0.842, p=0.002, 82.5% sensitivity and 56.2% specificity). miR-146a and miR-155 showed correlation with the diagnosis (rs=0.363 and 0.330, respectively) and the age of the patients (rs =0.239 and 0.366, respectively), and miR-155 showed correlation with the presence of secondary Raynaud syndrome (Spearman correlation coefficient=0.250) Conclusions: Our data showed that the expression levels of miR-146a and miR-155 in PB could be used as diagnostic biomarkers for SLE patients but larger study is needed to confirm these results. Key words: peripheral blood, miRNA, expression, systemic lupus erythematosus, biomarker.

Acute necrotizing retinal vasculitis as onset of systemic lupus erythematosus: A case report.

RATIONALE: Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by autoantibody production, complement activation, and deposition of immune complexes in tissues and organs. SLE can involve any region of the visual system. Although ocular manifestations are not part of the classification criteria for SLE, they can be observed in up to one-third of the patients with SLE. They are rarely reported at the time of disease onset. Retinal vasculitis is usually associated with active generalized disease. Due to its low frequency, we report a case of acute necrotizing retinal vasculitis as onset of SLE. PATIENT CONCERNS AND DIAGNOSIS: A 25-year-old white female was referred to the rheumatology clinic with gradually and rapid deterioration of the vision due to abnormal vessel permeability in the right fundus with edema along the vessels, occlusion of arterial branches in the middle periphery with leakage of the dye in these areas and indentical but less prominent changes with cotton wool spots in the papillomacular area and extensive hemorrhages in the left eye. The onset of malar rash, arthralgias and positive antinuclear, anti-double stranded DNA, anti-ribosomal P and anti-ß2 glycoprotein I antibodies with decreased C4 complement levels, as well as the positive lupus-band test confirmed the diagnosis of SLE. INTERVENTIONS: Aggressive immunomodulating therapy with high-dose methylprednisolone, intravenous immunoglobulin, and cyclophosphamide was used for suppression of the disease activity followed by azathioprine as maintaince therapy. OUTCOMES: Substantial improvement and partial resorption of the vasculitic changes, including central retinal artery and vein, was achieved prominently in the left eye. The study was conducted in accordance with the Declaration of Helsinki and written informed consent was obtained from the patient. Because of this, there is no need to conduct special ethic review and the ethical approval is not necessary. LESSONS: Inclusion of ocular manifestations among the classification criteria for SLE would enable earlier establishment of the diagnosis and therapeutic interventions in some instances of SLE.

Angioid streaks in aagenaes syndrome.

Aagenaes syndrome, also called lymphoedema cholestasis syndrome 1 (LSC1), is characterized by neonatal intrahepatic cholestasis, often lessening and becoming intermittent with age and severe chronic lymphoedema, mainly affecting the lower extremities. The condition is autosomal recessively inherited, and the gene is located on chromosome 15q. The locus, LCS1, was mapped to a 6.6 cM region on chromosome 15. Angioid streaks are visible irregular crack-like dehiscences in bruch's membrane that are associated with atrophic degeneration of the overlying retinal pigment epithelium. Angioid streaks have been described to be associated with pseudoxanthoma elasticum, paget's disease, sickle-cell anaemia, acromegaly, Ehlers-Danlos syndrome, and diabetes mellitus, but also appear in patients without any systemic diseases. Patients with angioid streaks are generally asymptomatic, unless the lesions extend towards the foveola or develop complications such as traumatic bruch's membrane rupture or macular choroidal neovascularization.

Purification and properties of a new thermostable cyclodextrin glucanotransferase from Bacillus pseudalcaliphilus 8SB.

A new cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) from an alkaliphilic halotolerant Bacillus pseudalcaliphilus 8SB was studied in respect to its γ-cyclizing activity. An efficient conversion of a raw corn starch into only two types of cyclodextrins (ß- and γ-CD) was achieved by the purified enzyme. Crude enzyme obtained by ultrafiltration was purified up to fivefold by starch adsorption with a recovery of 62% activity. The enzyme was a monomer with a molecular mass 71 kDa estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and native PAGE. The CGTase exhibited two pH optima, at pH 6.0 and 8.0, and was at most active at 60 °C and pH 8.0. The enzyme retained more than 80% of its initial activity in a wide pH range, from 5.0 to 11.0. The CGTase was strongly inhibited by 15 mM Cu(2+), Fe(2+), Ag(+), and Zn(2+), while some metal ions, such as Ca(2+), Na(+), K(+), and Mo(7+), exerted a stimulating effect in concentration of 5 mM. The important feature of the studied CGTase was its high thermal stability: the enzyme retained almost 100% of its initial activity after 2 h of heating at 40-60 °C; its half-life was 2 h at 70 °C in the presence of 5 mM Ca(2+). The achieved 50.7% conversion of raw corn starch into 81.6% ß- and 18.4% γ-CDs after 24 h enzyme reaction at 60 °C and pH 8.0 makes B. pseudalcaliphilus 8SB CGTase industrially important enzyme for cyclodextrin production.

Isolation of novel alkaliphilic bacillus strains for cyclodextrin glucanotransferase production.

New alkaliphilic Bacillus producers of cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) were isolated from 17 Bulgarian alkaline and normal habitats (springs and soils) by three steps of a selection. None of the isolates obtained, producing CGTase, appeared to be thermophilic in character. One hundred and thirty-seven strains were estimated for CGTase activity by batch cultivation in a liquid alkaline medium. Twenty-seven of them had a detectable CGTase activity in their culture supernatants under the enzyme assay conditions, despite of the significant growth of all isolates. The phenotypic properties of three selected strains (20RF, 8SB and 24WE) were determined. They were aerobic endospore-forming Bacillus strains: two of them were obligated alkaliphiles (20RF and 8SB) and one, alkalitolerant (24WE). Both obligated alkaliphiles were further characterised by 16S rRNA analysis. According to the full 16S rRNA gene sequences obtained and deposited to the NCBI GenBank database, both isolated obligated alkaliphiles 20RF and 8SB were clustered into the group of alkaliphilic Bacillus species. The exhibited CGTase production by them (230-250 U ml(-1) for 20RF and 130-160 U ml(-1) for 8SB) defined these new isolates as promising producers of the enzyme, especially Bacillus sp. 8SB synthesising thermostable alkaline beta-CGTase. Both new enzymes from 20RF and 8SB Bacillus strains formed only two types of cyclodextrins, beta and gamma, which could be of interest for their easy separation and industrial production.