Apoptosis of keratinocytes and serum DNase I activity in patients with cutaneous lupus erythematosus: relationship with clinical and immunoserological parameters.

BACKGROUND: Dysregulation of apoptosis has an important role in the induction of autoimmunity. OBJECTIVE: To evaluate the influence of keratinocyte apoptosis and deoxyribonuclease I (DNase I) activity on the clinical and immunoserological parameters of cutaneous lupus erythematosus (CLE). METHODS: We studied 69 CLE patients (39 with discoid LE (DLE), 12 with subacute CLE (SCLE), 12 with acute and 6 with intermittent CLE). Thirty of sixty-nine patients fulfilled criteria for systemic LE (SLE). Apoptotic index (AI) was evaluated immunohistochemically in lesional and non-lesional, photoprotected skin. Serum DNase I activity, antichromatin and anti-ENA antibodies were measured by ELISA. Disease activity was determined by SLEDAI-2K, SLICC/ACR, CLASI and RCLASI. RESULTS: AI in lesions was higher than in non-lesional skin (P < 0.001). There was no difference in AI between CLE and SLE patients. Patients with SCLE had higher lesional AI than patients with DLE (P < 0.05). We found a positive correlation between the lesional AI with CLASI A (P < 0.05) and RCLASI D (P < 0.05). CLE and SLE patients had significantly lower DNase I activity than healthy controls (P < 0.001). Patients with normal DNase I activity and low AI had significantly lower CLASI A than patients with decreased DNase I activity and/or elevated AI (P < 0.05). CONCLUSIONS: Increased keratinocyte apoptosis characterizes lesions of all CLE forms, especially of SCLE. AI correlates with CLE markers of acute and chronic inflammation. Normal level of apoptosis and DNase I activity simultaneously reduce the level of acute inflammation in CLE. Serum DNase I activity and AI might be important biomarkers in the evaluation of CLE patients.

Spin Hall Effect and Origins of Nonlocal Resistance in Adatom-Decorated Graphene.

Recent experiments reporting an unexpectedly large spin Hall effect (SHE) in graphene decorated with adatoms have raised a fierce controversy. We apply numerically exact Kubo and Landauer-Büttiker formulas to realistic models of gold-decorated disordered graphene (including adatom clustering) to obtain the spin Hall conductivity and spin Hall angle, as well as the nonlocal resistance as a quantity accessible to experiments. Large spin Hall angles of ∼0.1 are obtained at zero temperature, but their dependence on adatom clustering differs from the predictions of semiclassical transport theories. Furthermore, we find multiple background contributions to the nonlocal resistance, some of which are unrelated to the SHE or any other spin-dependent origin, as well as a strong suppression of the SHE at room temperature. This motivates us to design a multiterminal graphene geometry which suppresses these background contributions and could, therefore, quantify the upper limit for spin-current generation in two-dimensional materials.

Libyan Thymus capitatus essential oil: antioxidant, antimicrobial, cytotoxic and colon pathogen adhesion-inhibition properties.

AIMS: In the present work, the Libyan wild-growing Thymus capitatus essential oil (EO) was evaluated for its biological properties. METHODS AND RESULTS: Carvacrol (68.19%) and thymol (12.29%) were found to be the main compounds of the oil. Antioxidant properties, determined by 2,2-diphenylpicrylhydrazyl (DPPH) assay, revealed that IC50 values were 119, 403 and 105 µg ml(-1) for oil, thymol and carvacrol respectively. Microdilution method showed strong antibacterial and especially antifungal potential. Tetrazolium (MTT) colorimetric assay indicated moderate cytotoxicity towards human cell lines MRC-5, HCT 116 and HT-29 (IC50 = 30-150 µg ml(-1)). In adhesion-inhibition assay oil and main compounds reduced adhesion of Escherichia coli and Listeria monocytogenes on colon cells HT-29 (51 and 39% of inhibition against L. monocytogenes and E. coli respectively). CONCLUSIONS: Essential oil of Th. capitatus showed moderate cytotoxic activity, together with excellent antimicrobial effect, in particular against fungi, and significant potential to reduce pathogen colonization in colon. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report that EO of Th. capitatus could protect against colonization of pathogens to colon epithelium. Thymus capitatus from Libya should be recognized as possible new source of natural antioxidants, antimicrobials as well as possible source of new chemotherapeutics.

Biocompatibility of new nanostructural materials based on active silicate systems and hydroxyapatite: in vitro and in vivo study.

AIM: To evaluate in vitro cytotoxicity and in vivo inflammatory response to new nanostructural materials based on active calcium silicate systems (CS) and hydroxyapatite (HA-CS). METHODOLOGY: Cytotoxicity of eluates of new nanostructural noncommercial materials CS and HA-CS, and MTA (White MTA, Angelus(®) Soluções Odontológicas, Londrina, Brazil) as a control, were tested using the MTT assay on MRC-5 cells. Eluates of set materials were tested in 100% and 50% concentrations, 24 h, 7 days and 21 days post-elution. The pH values were determined for undiluted eluates of set materials. Polyethylene tubes containing the test materials (CS, HA-CS, MTA) were implanted in subcutaneous tissue of Wistar rats. Histopathological examinations were conducted at 7, 15, 30 and 60 days after the implantation. Data were statistically analyzed using three-way and one-way anova Tukey's post hoc test as well as Kruskall-Wallis test with Dunn's post hoc test at α = 0.05. RESULTS: All materials significantly reduced cell viability; especially when undiluted eluates were used (P < 0.001). After 24 h elution, cell viability was 10 ± 1.8%, 49.5 ± 4.2% and 61 ± 7.4%, for MTA, and HA-CS, respectively. However, CS and HA-CS were significantly less toxic than the control material MTA (P < 0.05). Cytotoxicity could be at least partially attributed to pH kinetics over time. Dilution of eluates of all tested materials resulted in better cell survival. Histopathological examination indicated similar inflammatory reaction, vascular congestion and connective tissue integrity associated with CS, HA-CS and MTA at each observation period (P > 0.05). The only significant difference was found for capsule thickness, that is thicker capsule was associated with HA-CS compared to MTA at 60 days (P = 0.0039). HA-CS induced moderately thick capsules (median score 3, score range 2-3), whereas MTA resulted in thin capsule formation (median score 2, score range 1-3). CONCLUSIONS: Evaluation of cytotoxicity and inflammatory response indicated better biocompatibility of CS and HA-CS, in comparison with MTA (White MTA, Angelus(®) Soluções Odontológicas, Londrina, Brazil).

New nanostructural biomaterials based on active silicate systems and hydroxyapatite: characterization and genotoxicity in human peripheral blood lymphocytes.

AIM: To characterize and investigate the genotoxic effect of a new endodontic cement based on dicalcium- and tricalcium-silicate (CS) with hydroxyapatite (HA) on human lymphocytes. METHODOLOGY: Hydrothermal treatment was applied for synthesis of CS and HA. The final mixture HA-CS, with potential to be used in endodontic practice, is composed of CS (34%) and HA (66%). Human lymphocytes were incubated with HA, HA-CS and CS for 1 h, at 37 °C and 5% CO2. Cell viability was determined using the trypan blue exclusion assay. To evaluate the level of DNA damage comet assay (single cell gel electrophoresis) was performed. For the statistical analysis anova and Duncan's Post Hoc Test were used. RESULTS: The SEM analysis indicated that CS consisted mostly of agglomerates of several micrometers in size, built up from smaller particles, with dimensions between 117 and 477 nm. This is promising because dimensions of agglomerates are not comparable with channels inside the cell membranes, whereas their nano-elements provide evident activity, important for faster setting of these mixtures compared to MTA. Values of DNA damage obtained in the comet assay indicated low genotoxic risk of the new endodontic materials. CONCLUSIONS: The significantly improved setting characteristics and low genotoxic risk of the new material support further research.

Correlation of human papilloma virus infection with cytology, colposcopy and histopathological examination of the bioptic tissue in low- and high-grade intraepithelial lesions.

INTRODUCTION: It is now believed that the majority of cervical cancer is preceded by long-term infection with high-risk types of the human papilloma virus (HPV). The presence of HPV high-risk types (HR-HPV) in the cells of intraepithelial change multiplies the possibility of its progressive development to high-grade cervical precancer and invasive disease. AIM: This study examined the correlation of HPV infection with cytology, colposcopy, and histopathological examination of the bioptic tissue in low- and high-grade cervical lesions. MATERIALS AND METHODS: This research was conducted as a study section. Data collection was performed during a ten-year period, at the University Clinic of Gynecology and Obstetrics - Narodni Front in Belgrade (Serbia). The basic set included 1,927 patients. Colposcopy, cytology, histopathology, and HPV test verification was made in all patients. Statistical analysis was performed using the SPSS program, version 17.0. Contingency tables were used to assess the degree of correlation of variables and chi-square test was used to determine the level of statistical significance in this study. A p value < 0.05 was considered statistically significant. RESULTS: Among 1,927 women studied, 635 (32.95 %) had abnormal cytological findings and among these, 272 (42.83%) were HR-HPV positive. There was a statistical difference between colposcopic and cytological findings in patients with HR-HPV (x2 = 35.33, p = 0.000). There was also a statistically significant difference between histophatological and colposcopical findings in patients with HR-HPV (x2 = 10,171, p = 0.001). Only HR-HPV types 16 and 18 showed a statistical significance compared to histopathological findings, unlike other HR-HPV. An important finding was that the authors found an abnormal colposcopy in 93.30% patients with low-grade intraepithelial neoplasia and 68.05% patients with low-grade squamous intraepithelial lesion (LSIL) had normal cytology and was 70.15 % HR-HPV negative. CONCLUSION: The findings imply that among high-grade intraepithelial neoplasias, the authors found a high presence of HPV type 16 and 18, and a statistical significant presence of HPV 16 in low-grade intraepithelial neoplasia, unlike other HR-HPV types in low-grade intraepithelial findings. The authors found a significant statistical correlation with abnormal cytology and presence of HPV type 16 in both groups (LSIL and high-grade squamous intraepithelial lesion (HSIL). The authors also found an abnormal colposcopy in 93.30% of patients with low-grade intraepithelial neoplasia, while 68.05% of patients with LSIL had normal cytology and were HR-HPV negative in 70.15% of the cases.

Progressive multifocal leukoencephalopathy associated with mycophenolate mofetil treatment in a woman with lupus and CD4+ T-lymphocyte deficiency.

There is an increase in the number of patients with systemic lupus erythematosus (SLE) reported as developing progressive multifocal leukoencephalopathy (PML) while on intensive immunosuppressive therapy. A 39-year-old HIV-negative woman with a 10-year history of SLE presented with progressive left-side weakness while on maintenance therapy with oral prednisone and mycophenolate mofetil (MMF). On several occasions low CD4+ T-lymphocyte counts were found (68/µL). Brain magnetic resonance imaging (MRI) revealed a large lesion in the right subcortical fronto-parietal region and a smaller one in the left frontal subcortex, corresponding to the PML. In cerebrospinal fluid, polymerase chain reaction (PCR) for JC virus (JCV) was negative, but anti-JCV antibodies were highly positive. Diagnosis of probable PML was made and MMF was withdrawn. The patient's condition improved with marked reduction of left-side weakness and an increase in CD4(+) T-lymphocyte count (141/µL). Follow-up MRI showed regression of lesions and over the next 6 months the patient remained stable. In spite of the grave prognosis associated with PML, SLE patients can have an excellent outcome if immunosuppressants are discontinued as soon as the correct diagnosis is made. SLE patients with associated low CD4(+) T-lymphocyte counts should be monitored for the development of PML during immunosuppressive therapy in particular.

Comparison of electrolytic status (Na+, K+, Ca2+, Mg2+) in preterm and term deliveries.

PURPOSE OF INVESTIGATION: The objective of this study was to evaluate the electrolytic status of Na+, K+, Ca+, and Mg2+ in serum and red blood cells in idiopathic preterm and term deliveries. METHODS: The study included 105 pregnant women diagnosed with idiopathic premature delivery (study group) and 36 pregnant women with physiologically term delivery (controls). Samples of mother's blood were collected and analyzed for the level of electrolytes in the serum/plasma and red blood cells. RESULTS: Measured values of magnesium in red blood cells in the study group were far lower than physiological values, intracellular calcium levels were higher in the study group compared to levels measured in the controls. Sodium concentrations in cells were significantly lower in subjects with premature delivery. CONCLUSION: The magnesium intracellular level is the best representative value of magnesium in the body.

Anti-PR3 immune responses induce segmental and necrotizing glomerulonephritis.

Wegener's granulomatosis (WG) is a life-threatening autoimmune vasculitis that affects lungs, kidneys and other organs. A hallmark of WG is the presence of classic anti-neutrophil cytoplasmic antibodies (c-ANCA) against self-proteinase 3 (PR3). Little is known about the role of these antibodies and PR3-specific immune responses in disease development. In this study, we demonstrate that PR3-specific autoimmune responses are pathogenic in non-obese diabetic (NOD) mice with an impaired regulatory arm of the immune response. Immunization of autoimmunity prone NOD mice with rmPR3 (recombinant mouse PR3) in complete Freund's adjuvant (CFA) resulted in high levels of c-ANCA, without detectable disease development. However, when splenocytes from these immunized mice were transferred into immunodeficient NOD-severe combined immunodeficiency (SCID) mice, the recipient mice developed vasculitis and severe segmental and necrotizing glomerulonephritis. No disease developed in NOD-SCID mice that received splenocytes from the CFA-alone-immunized donors (controls), indicating that disease development depends upon PR3-specific immune responses. In contrast to the pathology observed in NOD-SCID mice, no disease was observed when splenocytes from rmPR3-immunized C57BL/6 mice were transferred into immunodeficient C57BL/6-RAG-1(-/-) mice, suggesting that complex and probably multi-genetic factors play a role in the regulation of disease development.

Protective effect of linalool, myrcene and eucalyptol against t-butyl hydroperoxide induced genotoxicity in bacteria and cultured human cells.

We studied the protective effect of monoterpenes myrcene, eucalyptol and linalool against t-butyl hydroperoxide (t-BOOH) induced genotoxicity in reverse mutation assay with Escherichia coli WP2 IC185 strain and its oxyR mutant IC202, and with the comet assay in human hepatoma HepG2 and human B lymphoid NC-NC cells. The monoterpenes were tested in concentration ranges 0.05-1.5 mg/plate and 0.01-1.0 microg/ml in bacteria and mammalian cells, respectively. Suppression of t-BOOH induced mutagenesis was detected only in IC202 strain, and correlated with the observed inhibition of lipid peroxidation by the three monoterpenes. Linalool and myrcene strongly suppressed t-BOOH induced mutagenesis. Eucalyptol, in addition to moderate suppression of t-BOOH induced mutagenesis, suppressed also spontaneous mutagenesis. In NC-NC cells linalool and myrcene reduced t-BOOH induced DNA damage by about 50% at 0.01 microg/ml, while eucalyptol was less efficient (about 50% reduction at 1.0 microg/ml). In HepG2 cells linalool and eucalyptol reduced DNA damage by 30% and 40%, respectively, while myrcene was ineffective. The repair of t-BOOH induced DNA damage, studied in HepG2 cells, was not affected by monoterpenes. The results indicate that linalool, eucalyptol and myrcene have substantial protective effect against oxidant induced genotoxicity, which is predominately mediated by their radical scavenging activity.

Anti-double stranded DNA and lupus syndrome induced by interferon-beta therapy in a patient with multiple sclerosis.

We present a 43-year-old woman with relapsing-remitting multiple sclerosis (MS) who developed lupus syndrome after 32 months of IFN-beta-1a therapy. She presented with malaise, myalgia, arthralgia and fever. Laboratory tests showed high erythrocyte sedimentation rate, anaemia and lymphopenia. Antibodies to double stranded DNA (dsDNA) of IgG, IgM and IgA classes were detected on Critidia luciliae. Additionally, high levels of anti-nucleosomal antibodies, low levels of anti-histone and anti-Ro/SSA antibodies were also found. Diagnosis of drug-induced SLE was established. Treatment with IFN-beta was discontinued and oral prednisone was started. Twelve weeks after cessation of IFN-beta therapy, the patient's symptoms completely resolved and autoantibodies disappeared. To the best of our knowledge, this is the first report of a patient with MS in whom treatment with IFN-beta induced lupus syndrome and antibodies to dsDNA and nucleosome.

Antiribosomal-P protein antibodies in a patient with systemic lupus erythematosus and non-Hodgkin's lymphoma: more than coincidental finding?

Patients with systemic lupus erythematosus (SLE) are at an increased risk of lymphomas, but mechanisms underlying this association are obscure. Recently, it has been shown that antiribosomal-P protein (anti-P) antibodies cross-react with phospholipids and enhance the production of cytokines which may influence lymphomagenesis. We report a 46-year-old woman who suffered high grade diffuse large B-cell non-Hodgkin's lymphoma (DLBCL) 28 months after the diagnosis of SLE. Development of lymphoma was associated with occurrence of serum monoclonal IgM, and pronounced prolongation of phospholipid-dependent clotting tests. Anti-P IgG antibodies were highly positive both on HEp-2 cells and in ELISA test. Anticardiolipin, anti-beta2 glycoprotein I, and antiprothrombin IgM antibodies have also been found in high concentrations. Complete remission of DLBCL and SLE, with normalisation of clotting tests, and disappearance of M component was achieved with administration of rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone. The progression of SLE to DLBCL associated with presence of anti-P antibodies has not been previously reported. This association may not be coincidental, but further investigations are required to confirm this hypothesis.

Tunnel barrier enhanced voltage signal generated by magnetization precession of a single ferromagnetic layer.

We report the electrical detection of magnetization dynamics in an Al/AlOx/Ni80Fe20/Cu tunnel junction, where a Ni80Fe20 ferromagnetic layer is brought into precession under ferromagnetic resonance conditions. The dc voltage generated across the junction by the precessing ferromagnet is enhanced about an order of magnitude compared to the voltage signal observed when the contacts in this type of multilayered structure are Ohmic. We discuss the relation of this phenomenon to magnetic spin pumping and speculate on other possible underlying mechanisms responsible for the enhanced electrical signal.

Clinical and serological follow-up of 71 patients with anti-mitochondrial type 5 antibodies.

Antimitochondrial M5 type antibodies (AMA M5) have been described in patients with antiphospholipid syndrome (APS), thrombocytopenia and autoimmune hemolytic anemia. The aim of this study was to describe the clinical and immunological characteristics of a series of patients with AMA M5. We analyzed 71 patients with AMA M5 seen consecutively at our centres during the last 8 years. The clinical and immunological characteristics of diseases expression were compared with 70 consecutive disease control patients without AMA M5. Compared with the control group, AMA M5 positive patients presented higher prevalence of false positive VDRL test (P < 0.001) and thrombocytopenia (P = 0.002) with lower levels of anti-beta2 glycoprotein I antibodies. In AMA M5 patients (56 female, 15 male) a heterogeneous group of disorders were found. Twenty-seven (38%) patients fulfilled the Sapporo criteria for the classification of the APS. Laboratory criteria were met in 55 (77.5%), and clinical criteria in 31 (43.7%) patients. Six patients initially presented with non-criteria features of APS during follow-up period developed APS. Younger patients with AMA M5 should be carefully observed for the development of APS, even in the absence of serological criteria, while elderly must be screened for monoclonal gammopathy and hematological disorders.

Serum lipids and anti-oxidized LDL antibodies in primary antiphospholipid syndrome.

OBJECTIVE: The link between specific antibodies and atherogenesis in primary antiphospholipid syndrome (PAPS) is less strong than for thrombosis, although clearly the two processes are related and thrombosis is the main complication of atherosclerosis, a process known as atherothrombosis. The aim of this study was to investigate the influence of serum lipid levels and anti-oxidized LDL (oxLDL) antibodies on the clinical features of 42 patients with PAPS (mean age 40.45+/-13.37; 32 women and 10 men), and to compare them with 47 control subjects (mean age 39.68+/-13.93; 33 women and 14 men). METHODS: Total cholesterol, HDL and triglyceride concentrations were determined by enzymatic methods. LDL was calculated according to the Friedwald formula. Anticardiolipin, anti-oxidized LDL and anti-Beta2glycoprotein I antibodies were detected by ELISA. RESULTS: A significant association was found between arterial events and triglyceride, LDL and cholesterol concentrations, but multivariate analysis showed that cholesterol concentrations were the most important predictor of arterial events (p=0.012). Cerebrovascular insults were the most significantly associated with cholesterol concentrations (p=0.011). Myocardial infarctions were more frequently present in patients more than 40 years of age (p=0.032). No significant association of the investigated parameters with venous thromboses was found. Recurrent abortions were not associated with the presence or concentrations of the investigated parameters. Although patients had increased concentrations of anti-oxLDL antibodies, no significant association was found between the titres of anti-oxLDL antibodies and clinical features of APS. CONCLUSION: In patients with PAPS, lipid concentrations are a better predictor for arterial events than anti-oxLDL antibodies.

Heart and renal failure in renovascular hypertension caused by giant cell arteritis--case report.

We report a case of a male teenager with severe heart and acute renal failure as the dominant clinical manifestations of renovascular hypertension (RVH) caused by atypical giant cell arteritis (GCA). Unrecognized RVH and treatment of the consequent heart failure by angiotensin-converting enzyme inhibitors (ACEI) probably contributed to progression of renovascular disease to bilateral renal artery occlusion. Recurrent "flash" pulmonary edemas could not be prevented until surgical revascularization of the only functioning right kidney was achieved by an aortorenal bypass. Prompt post-operative normalization of heart function and arterial hypertension occurred despite the histopathological finding of the resected renal artery compatible with GCA and 4-year duration of significant renovascular disease. At the last check-up, the patient was asymptomatic, with normal arterial pressure on the prescribed treatment: carvedilol, hydrochlorothiazide, prednisolone 20 mg daily and aspirin. Subsequent follow-up is necessary to observe the evolution of GCA as an exceptionally rare cause of RVH.

Rothmund-Thomson syndrome. The first case with plantar keratoderma and the second with coeliac disease.

We report two unusual patients with Rothmund-Thomson syndrome (RTS), a rare genodermatosis. The first patient is a 5-year-old girl with congenital poikiloderma, photosensitivity, plantar punctate keratoderma, stunted growth and severe mental retardation. Plantar keratoderma associated with RTS has been reported only once. The second patient is a 21-year-old female presenting with rounded "moon" face, trunk obesity, coeliac disease, short stature and mild mental retardation. This is the first case of RTS associated with coeliac disease.

Ovarian neoplasms and pregnancy.

The aim of our study was to investigate the incidence of ovarian malignant neoplasms in pregnancy. In the prospective study, during a 2-year period of time, we had the ethical dilemma concerning the conciliar treatment vs the patient's wish to deliver and save the reproductive capabilities. We examined 37 pregnant women; the incidence of ovarian cancer was 13.5% vs 6.5% in other works. Diagnoses were made by ultrasound criteria and physical examinations. Statistically significant results were obtained by Student t test. Mean gestational age was 20.1 weeks and mean age 31.1 years. Treatment depends on the neoplasms type, grade, and presence of the metastatic pathways. In benign neoplasms, we used laparoscopic treatment with minor invasion. Distribution of benign neoplasms was in the same range as that in other works.

Choriocarcinoma--postdisease ultrasonographic findings.

This is a case report of consequences that malignant gestational trophoblastic disease (GTD) can cause on reproductive health protruding into uterine wall and damaging uterine tissue. Transvaginal Doppler ultrasound examination can be of great value in detecting molar tissue, protrusion of malignant trophoblast in uterine wall, and neovasularization in malignant tissue. It is expected to measure a low resistance index in a field of neovascularization, because neovascularization in malignancy is not rare and those vessels do not have muscular stratum. This case is an example of possible irreversible serious and large damages that can be seen after successful treatment of GTD. They are warning on possible high degree of malignancy in GTD as well as on possible serious impact on reproductive health.