Expression of BAFF receptors in muscle tissue of myositis patients with anti-Jo-1 or anti-Ro52/anti-Ro60 autoantibodies

Introduction

Anti-Jo-1 and anti-Ro52 autoantibodies are common in patients with myositis, but the mechanisms behind their production are not known. Survival of autoantibody-producing cells is dependent on B-cell-activating factor of the tumour necrosis factor family (BAFF). BAFF levels are elevated in serum of anti-Jo-1-positive myositis patients and are influenced by type-I interferon (IFN). IFN-producing cells and BAFF mRNA expression are present in myositis muscle. We investigated expression of the receptors for BAFF in muscle tissue in relation to anti-Jo-1 and anti-Ro52/anti-Ro60 autoantibodies and type-I IFN markers.

Methods

Muscle biopsies from 23 patients with myositis selected based on autoantibody profile and 7 healthy controls were investigated for expression of BAFF receptor (BAFF-R), B-cell maturation antigen (BCMA) and transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI). Nineteen samples were assessed for plasma (CD138) and B-cell (CD19) markers. The numbers of positive cells per area were compared with the expression of plasmacytoid dendritic cell (pDC) marker blood dendritic cell antigen-2 (BDCA-2) and IFNα/β-inducible myxovirus resistance-1 protein (MX-1).

Results

BAFF-R, BCMA and TACI were expressed in five, seven and seven patients, respectively, and more frequently in anti-Jo-1-positive and/or anti-Ro52/anti-Ro60-positive patients compared to controls and to patients without these autoantibodies (P = BAFF-R: 0.007, BCMA: 0.03 and TACI: 0.07). A local association of receptors with B and plasma cells was confirmed by confocal microscopy. The numbers of CD138-positive and BCMA-positive cells were correlated (r = 0.79; P = 0.001). Expression of BDCA-2 correlated with numbers of CD138-positive cells and marginally with BCMA-positive cells (r = 0.54 and 0.42, respectively; P = 0.04 and 0.06, respectively). There was a borderline correlation between the numbers of positively stained TACI cells and MX-1 areas (r = 0.38, P = 0.08).

Conclusions

The expression pattern of receptors for BAFF on B and plasma cells in muscle suggests a local role for BAFF in autoantibody production in muscle tissues of patients with myositis who have anti-Jo-1 or anti-Ro52/anti-Ro60 autoantibodies. BAFF production could be influenced by type-I IFN produced by pDCs. Thus, B-cell-related molecular pathways may participate in the pathogenesis of myositis in this subset of patients.

Electronic supplementary material

The online version of this article (doi:10.1186/s13075-014-0454-8) contains supplementary material, which is available to authorized users.     

The interrelations of radiologic findings and mechanical ventilation in community acquired pneumonia patients admitted to the intensive care unit: a multicentre retrospective study

Background

Burkholderia cepacia complex (BCC) bacteria are highly virulent, typically multidrug-resistant, opportunistic pathogens in cystic fibrosis (CF) patients and other immunocompromised individuals. B. vietnamiensis is more often susceptible to aminoglycosides than other BCC species, and strains acquire aminoglycoside resistance during chronic CF infection and under tobramycin and azithromycin exposure in vitro, apparently from gain of antimicrobial efflux as determined through pump inhibition. The aims of the present study were to determine if oxidative stress could also induce aminoglycoside resistance and provide further observations in support of a role for antimicrobial efflux in aminoglycoside resistance in B. vietnamiensis.

Findings

Here we identified hydrogen peroxide as an additional aminoglycoside resistance inducing agent in B. vietnamiensis. After antibiotic and hydrogen peroxide exposure, isolates accumulated significantly less [³H] gentamicin than the susceptible isolate from which they were derived. Strains that acquired aminoglycoside resistance during infection and after exposure to tobramycin or azithromycin overexpressed a putative resistance-nodulation-division (RND) transporter gene, amrB. Missense mutations in the repressor of amrB, amrR, were identified in isolates that acquired resistance during infection, and not in those generated in vitro.

Conclusions

These data identify oxidative stress as an inducer of aminoglycoside resistance in B. vietnamiensis and further suggest that active efflux via a RND efflux system impairs aminoglycoside accumulation in clinical B. vietnamiensis strains that have acquired aminoglycoside resistance, and in those exposed to tobramycin and azithromycin, but not hydrogen peroxide, in vitro. Furthermore, the repressor AmrR is likely just one regulator of the putative AmrAB-OprM efflux system in B. vietnamiensis.     

Centrosome-declustering drugs mediate a two-pronged attack on interphase and mitosis in supercentrosomal cancer cells

Classical anti-mitotic drugs have failed to translate their preclinical efficacy into clinical response in human trials. Their clinical failure has challenged the notion that tumor cells divide frequently at rates comparable to those of cancer cells in vitro and in xenograft models. Given the preponderance of interphase cells in clinical tumors, we asked whether targeting amplified centrosomes, which cancer cells carefully preserve in a tightly clustered conformation throughout interphase, presents a superior chemotherapeutic strategy that sabotages interphase-specific cellular activities, such as migration. Herein we have utilized supercentrosomal N1E-115 murine neuroblastoma cells as a test-bed to study interphase centrosome declustering induced by putative declustering agents, such as Reduced-9-bromonoscapine (RedBr-Nos), Griseofulvin and PJ-34. We found tight ‘supercentrosomal'' clusters in the interphase and mitosis of ~80% of patients'' tumor cells with excess centrosomes. RedBr-Nos was the strongest declustering agent with a declustering index of 0.36 and completely dispersed interphase centrosome clusters in N1E-115 cells. Interphase centrosome declustering caused inhibition of neurite formation, impairment of cell polarization and Golgi organization, disrupted cellular protrusions and focal adhesion contacts—factors that are crucial prerequisites for directional migration. Thus our data illustrate an interphase-specific potential anti-migratory role of centrosome-declustering agents in addition to their previously acknowledged ability to induce spindle multipolarity and mitotic catastrophe. Centrosome-declustering agents counter centrosome clustering to inhibit directional cell migration in interphase cells and set up multipolar mitotic catastrophe, suggesting that disbanding the nuclear–centrosome–Golgi axis is a potential anti-metastasis strategy.Unlike in vitro cell cultures, cancer cells in patients'' tumor tissues have low mitotic indices and proliferation rates.¹ Consequently, drugs targeting mitosis demonstrate limited clinical efficacy, which exposes a fundamental weakness in the rationale underlying their clinical development. By contrast, classical microtubule-targeting agents (MTAs), largely believed to act by perturbing mitosis, remain the mainstay of chemotherapy in the clinic. Given the miniscule population of mitotic cells in patient tumors,^{2, 3} it stands to reason that MTAs must target interphase.⁴ This paradigm shift has spurred an intense search for novel interphase targets that combine the ‘ideal'' attributes of cancer-cell selectivity and the ability to confer vulnerability on a large proportion of tumor cells.Centrosomes, the major microtubule-organizing centers (MTOCs) of cells, are required for accurate cell division, cell motility and cilia formation.⁵ The number of centrosomes within a cell is strictly controlled, and their duplication occurs only once per cell cycle. Nearly all types of cancer cells have abnormal numbers of centrosomes,^{6, 7, 8} which correlates with chromosomal instability during tumorigenesis.^{9, 10, 11} Supernumerary centrosomes in cancer cells can cause spindle multipolarity and thus non-viable progeny. Cancer cells avoid this outcome by clustering centrosomes to assemble a pseudo-bipolar mitotic spindle, which yields viable daughter cells.¹² Thus disrupting centrosome clustering may selectively drive cancer cells with amplified centrosomes to mitotic catastrophe and apoptosis without affecting normal cells.The fate and interphase role of the supercentrosomal cluster inherited by each daughter cell at the end of a pseudobipolar mitosis is unknown. This is an important research question, because a majority of cells within tumors are in interphase and the centrosomes'' command over microtubule nucleation is crucial for the cellular organization and motility in interphase. If cancer cells cluster centrosomes in interphase, then disrupting the cluster could impact interphase-specific processes, opening up a vital therapeutic avenue. We envision that centrosome declustering would (a) derail interphase-specific polarization and migration processes and (b) precipitate multipolar mitosis culminating in apoptosis. This two-pronged strategy would impact a significantly larger proportion of tumor cells and consign them to death. Our study herein establishes that centrosome-declustering drugs (RedBr-Nos, Griseofulvin and PJ-34) achieve this two-pronged attack as a unique class of agents that exhibit multiple cellular activities.     

Accumulation of weathered pp'-DDE in xylem sap of grafted watermelon

Movement of weathered p,p'-dichlorodiphenyldichloroethane (p,p'-DDE) from contaminated soil to the rhizosphere pore water to the xylem sap of grafted watermelon was studied under green house conditions. p,p'-DDE concentrations in pore water and xylem sap was compared in intact plants, homografted, and compatible heterografts of Cucurbita pepo spp. pepo and Citrullus lanatus plants. An average p,p'-DDE concentrations in pore water of contaminated soil ranged from 0.36 microg/L to 0.55 microg/L and there were no statistically significant among the cultivars. Conversely, the xylem sap p,p'-DDE concentration of heterografted watermelon having a zucchini rootstock and watermelon scion was 71 microg/L and it was greater than intact watermelon plants (0.49 microg/L) but less than that of intact plants of zucchini (141 microg/L). Homografting showed no effect on xylem sap p,p'-DDE concentrations of the identical cultivars. The bio-concentration factors (BCFs) which is an average p,p'-DDE concentration in xylem sap over average p,p'-DDE in pore water were 344, 325, 197, 1.28, and 0.89 for intact plant of zucchini, homografted zucchini, heterografted watermelon, homografted watermelon, and intact plant of watermelon, respectively. Xylem sap p,p'-DDE concentrations of the heterografted watermelon plants were clearly influenced by plant phylogeny and enhanced by the zucchini rootstock compared to intact watermelon plants.     

Histological and enzyme histochemical investigation of the hemal nodes of the hair goat

We investigated the structure of the hemal node in six healthy hair goats using histological and enzyme histochemical methods. After processing, tissue sections were stained with Crossman's trichrome, Gordon-Sweet's silver and Pappenheim's panoptic stains. Alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACP-ase) were demonstrated in frozen sections. Hemal nodes were encapsulated by connective tissue and few smooth muscle cells. Several trabeculae originated from the capsule and extended into the hemal node. A subcapsular sinus was present beneath the capsule and was continuous with the deeper sinuses. Subcapsular and deep sinuses were filled with erythrocytes. The parenchyma consisted of lymphoid follicles, diffuse interfollicular lymphocytes and irregular wide lymphoid cords. Cortical and medullary regions were not distinct. ANAE (+) and ACP-ase (+) cells were located mainly in the germinal centers of the lymphoid follicles and also were scattered equally in the interfollicular region and lymphoid cords. Monocytes, macrophages and reticular cells displayed a diffuse positive reaction, whereas localized granular positivity was observed in lymphocytes. We demonstrated that the general structure of the hair goat hemal nodes is similar to that of other ruminant species.     

Lymphocyte DNA damage and oxidative stress in patients with iron deficiency anemia

Oxidant stress has been shown to play an important role in the pathogenesis of iron deficiency anemia. The aim of this study was to investigate the association between lymphocyte DNA damage, total antioxidant capacity and the degree of anemia in patients with iron deficiency anemia. Twenty-two female with iron deficiency anemia and 22 healthy females were enrolled in the study. Peripheral DNA damage was assessed using alkaline comet assay and plasma total antioxidant capacity was determined using an automated measurement method. Lymphocyte DNA damage of patients with iron deficiency anemia was significantly higher than controls (p<0.05), while total antioxidant capacity was significantly lower (p<0.001). While there was a positive correlation between total antioxidant capacity and hemoglobin levels (r=0.706, p<0.001), both total antioxidant capacity and hemoglobin levels were negatively correlated with DNA damage (r=-0.330, p<0.05 and r=-0.323, p<0.05, respectively). In conclusion, both oxidative stress and DNA damage are increased in IDA patients. Increased oxidative stress seems as an important factor that inducing DNA damage in those IDA patients. The relationships of oxidative stress and DNA damage with the severity of anemia suggest that both oxidative stress and DNA damage may, in part, have a role in the pathogenesis of IDA.     

Adsorption of Cd(II) ions from aqueous solutions using activated carbon prepared from olive stone by ZnCl2 activation

This study is aimed to remove Cd(II) ions from aqueous solutions by adsorption. As adsorbent, activated carbon prepared from olive stone, an agricultural solid by-product was used. Different activating agent (ZnCl(2)) amounts and adsorbent particle size were studied to optimize adsorbent surface area. The adsorption experiments were conducted at different parameters such as, adsorbent dose, temperature, equilibrium time and pH. According to the experiments results, the equilibrium time, optimum pH, adsorbent dosage were found 60 min, pH > 6 and 1.0 g/50 ml respectively. The kinetic data supports pseudo-second order model and intra-particle model but shows very poor fit for pseudo-first order model. Adsorption isotherms were obtained from three different temperatures. These adsorption data were fitted with the Langmuir and Freundlich isotherms. In addition, the thermodynamic parameters, standard free energy (DeltaG(0)), standard enthalpy (DeltaH(0)), standard entropy (DeltaS(0)) of the adsorption process were calculated. To reveal the adsorptive characteristics of the produced active carbon, BET surface area measurements were made. Structural analysis was performed using SEM-EDS. The resulting activated carbons with 20% ZnCl(2) solution was the best sample of the produced activated carbons from olive stone with the specific surface area of 790.25 m(2)g(-1). The results show that the produced activated carbon from olive stone is an alternative low-cost adsorbent for removing Cd(II).     

The effects of heat treatment on physical properties and surface roughness of red-bud maple (Acer trautvetteri Medw.) wood

Heat treatment is often used to improve the dimensional stability of wood. In this study, the effects of heat treatment on physical properties and surface roughness of red-bud maple (Acer trautvetteri Medw.) wood were examined. Samples obtained from Düzce Forest Enterprises, Turkey, were subjected to heat treatment at varying temperatures and durations. The physical properties of heat-treated samples were compared against controls in order to determine their; oven-dry density, air-dry density, and swelling properties. A stylus method was employed to evaluate the surface characteristics of the samples. Roughness measurements, using the stylus method, were made in the direction perpendicular to the fiber. Three main roughness parameters; mean arithmetic deviation of profile (Ra), mean peak-to-valley height (Rz), and maximum roughness (Rmax) obtained from the surface of wood, were used to evaluate the effect of heat treatment on the surface characteristics of the specimens. Significant differences were determined (p>0.05) between surface roughness parameters (Ra, Rz, Rmax) at three different temperatures and three periods of heat treatment. The results showed that the values of density, swelling and surface roughness decreased with increasing temperature treatment and treatment times. Red-bud maple wood could be utilized successfully by applying proper heat treatment techniques without any losses in investigated parameters. This is vital in areas, such as window frames, where working stability and surface smoothness are important factors.     

Lymphatic mapping and sentinel node biopsy in gynecological cancers: a critical review of the literature

Although it does not have a long history of sentinel node evaluation (SLN) in female genital system cancers, there is a growing number of promising study results, despite the presence of some aspects that need to be considered and developed. It has been most commonly used in vulvar and uterine cervivcal cancer in gynecological oncology. According to these studies, almost all of which are prospective, particularly in cases where Technetium-labeled nanocolloid is used, sentinel node detection rate sensitivity and specificity has been reported to be 100%, except for a few cases. In the studies on cervical cancer, sentinel node detection rates have been reported around 80–86%, a little lower than those in vulva cancer, and negative predictive value has been reported about 99%. It is relatively new in endometrial cancer, where its detection rate varies between 50 and 80%. Studies about vulvar melanoma and vaginal cancers are generally case reports. Although it has not been supported with multicenter randomized and controlled studies including larger case series, study results reported by various centers around the world are harmonious and mutually supportive particularly in vulva cancer, and cervix cancer. Even though it does not seem possible to replace the traditional approaches in these two cancers, it is still a serious alternative for the future. We believe that it is important to increase and support the studies that will strengthen the weaknesses of the method, among which there are detection of micrometastases and increasing detection rates, and render it usable in routine clinical practice.     

Genotoxicity testing of fluconazole in vivo and in vitro

The genotoxic effects of the antifungal drug fluconazole (trade name triflucan) were assessed in the chromosome aberration (CA) test in mouse bone-marrow cells in vivo and in the chromosome aberration, sister chromatid exchange (SCE) and micronucleus (MN) tests in human lymphocytes. Fluconazole was used at concentrations of 12.5, 25.0 and 50.0 mg/kg for the in vivo assay and 12.5, 25.0 and 50.0 microg/ml were used for the in vitro assay. In both test systems, a negative and a positive control (MMC) were also included. Six types of structural aberration were observed: chromatid and chromosome breaks, sister chromatid union, chromatid exchange, fragments and dicentric chromosomes. Polyploidy was observed in both the in vivo and in vitro systems. In the in vivo test, fluconazole did not significantly increase the frequency of CA. In the in vitro assays, CA, SCE and MN frequencies were significantly increased in a dose-dependent manner compared with the negative control. The mitotic, replication and cytokinesis-block proliferation indices (CBPI) were not affected by treatments with fluconazole. According to these results, fluconazole is clastogenic and aneugenic in human lymphocytes, but these effects could not be observed in mice. Further studies should be conducted in other test systems to evaluate the full genotoxic potential of fluconazole.