The role of neutrophil activation in determining the outcome of pregnancy and modulation by hormones and/or cytokines.

Neutrophils are often exclusively considered as a first-line innate immune defence, able to rapidly kill or trap pathogens and causing in case of over-activation tissue damage. In the female reproductive tract, however, the presence and activity of neutrophils seems to be tightly regulated. Major players in orchestrating this regulation are cyclical steroid sex hormones present during the menstrual cycle and pregnancy. This review describes the role of sex hormones in regulating directly or indirectly the functionality of neutrophils, the role of neutrophils during fertilization and pregnancy and in controlling viral, fungal and bacterial infection. This review also discusses the consequence of overt neutrophil activation in pregnancy pathologies.

Refractory neuro-Behçet treated by tocilizumab: a case report.

A patient with central nervous system involvement of Behçet's disease was refractory to conventional immunosuppressive therapy and showed secondary failure of the anti-TNF agent infliximab. This presented as a progressive weakness of the legs and reduction in walking distance. The cerebrospinal fluid showed signs of inflammation including a vastly elevated IL-6 concentration. Given this result, the anti-IL-6 receptor antibody tocilizumab was administered and a good improvement of inflammatory parameters and a satisfactory increase of the walking distance were achieved.

[Comparison of laser treatment of patients with the replace with "computer-based laser" between experienced and unexperienced operators]. / Vergleich der laserbehandlung mit einem computergestützten lasersystem durch einen erfahrenen versus einen unerfahrenen behandelnden arzt.

INTRODUCTION: The aim of this study was to compare individual accuracy and speed of laser application using the Navilas laser system in patients with diabetic and vascular macular oedema. METHODS: The Navilas laser system was used by an experienced retina specialist and a relatively inexperienced ophthalmologist. Obtained results were retrospectively assessed with regard to precision and speed of laser application. RESULTS: Precision of laser therapy was quite similar, 88 % and 84 % for experienced and inexperienced operators, respectively. The treatment procedure took on average 5.5 minutes and 5.34 minutes for experienced and inexperienced operators, respectively. CONCLUSIONS: Precision and duration of therapy using the Navilas laser system do not seem to be correlated with the individual experience of the treating physician.

Comparative effectiveness of antitumour necrosis factor agents, biologics with an alternative mode of action and tofacitinib in an observational cohort of patients with rheumatoid arthritis in Switzerland.

BACKGROUND: Multiple biologic and targeted synthetic disease-modifying rheumatic drugs (b/tsDMARDs) are approved for the management of rheumatoid arthritis (RA), including TNF inhibitors (TNFi), bDMARDs with other modes of action (bDMARD-OMA) and Janus kinase inhibitors (JAKi). Combination of b/tsDMARDs with conventional synthetic DMARDs (csDMARDs) is recommended, yet monotherapy is common in practice. OBJECTIVE: To compare drug maintenance and clinical effectiveness of three alternative treatment options for RA management. METHODS: This observational cohort study was nested within the Swiss RA Registry. TNFi, bDMARD-OMA (abatacept or anti-IL6 agents) or the JAKi tofacitinib (Tofa) initiated in adult RA patients were included. The primary outcome was overall drug retention. We further analysed secondary effectiveness outcomes and whether concomitant csDMARDs modified effectiveness, adjusting for potential confounding factors. RESULTS: 4023 treatment courses of 2600 patients were included, 1862 on TNFi, 1355 on bDMARD-OMA and 806 on Tofa. TNFi was more frequently used as a first b/tsDMARDs, at a younger age and with shorter disease duration. Overall drug maintenance was significantly lower with TNFi compared with Tofa [HR 1.29 (95% CI 1.14 to 1.47)], but similar between bDMARD-OMA and Tofa [HR 1.09 (95% CI 0.96 to 1.24)]. TNFi maintenance was decreased when prescribed without concomitant csDMARDs [HR: 1.27 (95% CI 1.08 to 1.49)], while no difference was observed for bDMARD-OMA or Tofa maintenance with respect to concomitant csDMARDs. CONCLUSION: Tofa drug maintenance was comparable with bDMARDs-OMA and somewhat higher than TNFi. Concomitant csDMARDs appear to be required for optimal effectiveness of TNFi, but not for bDMARD-OMA or Tofa.

Long-term immunogenicity after yellow fever vaccination in immunosuppressed and healthy individuals.

BACKGROUND: The live-attenuated yellow fever vaccine (YFV) is generally contraindicated in immunosuppressed patients. Our aim was to investigate if immunosuppressive therapy impairs the long-term protection against yellow fever virus in patients who had received YFV prior to the start of their immunosuppressive therapy. METHODS: Our study examined 35 healthy individuals and 40 immunosuppressed patients with autoimmune diseases or organ transplants. All individuals had received YFV prior to the onset of their immunosuppression. We analysed the long-term influence of the immunosuppressive therapy on the YFV protective immunity by measuring neutralising antibodies (NA) with the Plaque Reduction Neutralisation Test (PRNT). We assessed risk factors for a negative PRNT result (titre below 1: 10) and their influence on the magnitude of the NA. RESULTS: A median time interval of 21.1 years (interquartile range 14.4-31.3 years) after the YFV in all patients, a total of 35 immunosuppressed patients (88%) were seropositive (PRNT ≥ 1:10) compared to 31 patients (89%) in the control group. The geometric mean titres of NA did not differ between the groups. The duration of an underlying rheumatic disease was the only risk factor found for a lower magnitude of NA. An insufficient level of NA was found in nine subjects (12%) who had received a single dose of YFV (in one subject, the number of YFV doses was unknown). CONCLUSION: The use of an immunosuppressive drug started after the administration of the YFV did not affect long-term persistence of NA. A second dose of YFV may be necessary to secure long-term immunity.

Zygnematalean zygospores: morphological features and use in species identification.

Suitable morphological characteristics for identification of zygnematalean algae were determined using a combination of classical light microscopy (LM) techniques, fluorescence microscopy (with blue and green excitation), scanning electron microscopy (SEM) and specialized culture methods. Characteristics of spore germination, growth and reproduction under culture conditions identified Zygnema chalybeo-spermum in a mixture of zygnematalean spores collected from a small fishpond in Czechia. Reproduction in general, particularly aplanospore formation and lateral conjugation was more frequent and occurred earlier in a nutrient poor medium than in a nutrient rich medium, where vegetative growth was more vigorous. Variability in spore size was wide and influenced by the origin (sexual and/or asexual) of the spores. Asexual spores, particularly partenospores were rounded and significantly smaller than sexual ones. Thus spore morphology alone (size and shape) is not a particularly helpful characteristic for species identification. The mesospore of mature spores contained lipids and a sporopolenin-like material (algaenan), which showed green autofluorescence with blue excitation. The mesospore ornamentation, the only characteristic found that is suitable for identification purposes, can be observed easily in LM and SEM after exospore removal by KOH treatment. Detailed LM and SEM observations of the zygospores of all Zygnema species thus could provide basic data necessary for the preparation of an atlas and key for species identification which, after completion with molecular methods, brings clarification into the genetic relationships between morphospecies.

B cell receptor signaling and autoimmunity.

The immune receptors of lymphocytes are able to sense the nature of bound ligands. Through coupled signaling pathways the generated signals are appropriately delivered to the intracellular machinery, allowing specific functional responses. A central issue in contemporary immunology is how the fate of B lymphocytes is determined at the successive developmental stages and how the B cell receptor distinguishes between signals that induce immune response or tolerance. Experiments with mice expressing transgenes or lacking signal transduction molecules that lead to abnormal lymphocyte development and/or response are providing important clues to the mechanisms that regulate signaling thresholds at different developmental stages. The studies are also revealing novel potential mechanisms of induction of autoimmunity, which may have a bearing on the understanding of human diseases.

In vitro and in vivo evaluation of iodine-123-Ro 16-0154: a new imaging agent for SPECT investigations of benzodiazepine receptors.

The flumazenil analogue, Ro 16-0154, a benzodiazepine partial inverse agonist, has been labeled by halogen exchange to enable SPECT investigations of central benzodiazepine receptors in the human brain. The purified 123I-Ro 16-0154 was found to be stable in rat brain preparations and to be metabolized in rat liver preparations. Its pharmacologic properties were comparable to those of flumazenil. The biodistribution in rats (1 hr postinjection) resulted in a high brain-to-blood ratio of 16. Clinical studies revealed images of the benzodiazepine receptor density in the brain. Since the receptor labeling was markedly reduced by injection of flumazenil, it was considered to be specific. Storage defects due to pathologic cerebral blood flow and changed receptor density were detected; this shows the potential usefulness of the substance for diagnostic purposes, e.g., the differential diagnosis of various forms of epilepsy.

Vasculitides in hairy cell leukemia.

Forty-two cases of vasculitis coincident with hairy cell leukemia (HCL) have been reported, of which 17 had panarteritis nodosa (PAN), 21 had cutaneous leukocytoclastic vasculitis (LCV), and 4 had vessel wall infiltration by hairy cells. PAN generally occurred after the diagnosis of HCL, splenectomy, and infection. HBs antigen was detected in 3 of 12 patients tested, whereas immune complexes were positive in 3 of 4 patients tested. LCV was often preceded by infection and was frequently detected before HCL. Serum immunoglobulin levels were generally elevated when measured. Cryoglobulins, complement, rheumatoid factor, and antinuclear antibodies showed no clear association with vasculitis in HCL. These reports suggest a role for infection and splenectomy as contributing factors to vasculitis.

123I-labeling and evaluation of Ro 43-0463, a SPET tracer for MAO-B imaging.

Using the copper assisted halogen exchange the MAO-B inhibitor Ro 43-0463, N-(2-aminoethyl)-5-iodo-2-pyridinecarboxamide, was labelled with 123I as well as with 125I to allow in vitro and in vivo investigations including SPET with healthy volunteers. Ro 43-0463 is known to inhibit reversibly and specifically MAO-B, having an IC50 of 3 x 10(-8) Mol/L. The labeling in the presence of CuSO4 and ascorbic acid was optimised, varying time (30 to 105 min), precursor concentration (1-3.5 mg) and temperature (130-200 degrees C). The labeling yield ranged between 60 and 70%. Purification was achieved with Lichrosorb RP-18 (5 micron, 250 x 8 mm) and 1.5 mL/min 0.36 M H3PO4/EtOH 97/3 [0.01 M (NH4)2HPO4]. After neutralisation and sterile filtration the final activity concentration ranged between 18.5 and 37 MBq/mL. Biodistribution studies showed a brain to blood ratio greater than 1 within 1 h p.i. The main radiation burden calculated from these animal data is to alimentary and excretory organs and the ovaries. Autoradiography was performed using rat brain slices and 5 nM [125I]Ro 43-0463 in TRIS-buffer pH 7.4 for 90 min at 20 degrees C. Its radioactivity pattern corresponds to the known distribution of MAO-B in the rat brain. By displacement with L-deprneyl the highly specific binding of R0 43-0463 was proven in vitro. SPECT studies with normal volunteers corresponded with the pattern found in autoradiography.

In vitro and in vivo testing of the dopamine D1 ligand [123I]SCH 23982 with respect to its potential application in SPET investigations.

[123I]SCH 23982, a dopamine D1 ligand, was labelled in a large scale process and then tested in vitro for binding to rat brain sections and membranes. Because of the promising values of KD = 1.5 x 10(-10) M and Bmax = 0.7 x 10(-11) mol/g, in vivo evaluation was performed on rats and normal volunteers to test its possible usefulness for SPET imaging. In competition experiments, a higher binding in the presence of sulpiride was found while ketanserin displaced [123I]SCH 23982 only at a 10,000-fold excess. Differences between rats and men were seen with respect to their metabolism. SPET investigations failed because the washout of [123I]SCH 23982 was too rapid.

[76Br]Bromodeoxyuridine, a potential tracer for the measurement of cell proliferation by positron emission tomography, in vitro and in vivo studies in mice.

5-Bromo-2'-deoxyuridine (BrUdR) labeled with 77Br and 76Br was compared with 5-iodo-2'-deoxyuridine (IUdR) labeled with 125I or 131I, first in vitro then in in vivo experiments in mice. The results showed a significantly higher incorporation of BrUdR into DNA than IUdR, which can be explained by the greater similarity (size and surface hydrophilicity of the molecules) of BrUdR to thymidine. Both tracers are dehalogenated quickly in vivo but not in vitro. Free bromide is excreted more slowly than iodide, resulting in a higher background activity level after the application of [76Br]BrUdR and compensates for the favorable DNA incorporation. 76Br has more favorable properties than 124I for imaging purposes with positron emission tomography (PET) because of a very convenient half-life (16 h vs. 4.15 days) and about double the positron yield per decay. However, the more favorable physical properties are balanced by the slower excretion and thus the estimated radiation dose is higher in the case of 76Br than 124I. Thus, both tracers, [124I]IUdR and [76Br]BrUdR are potentially suitable but not optimal to measure cell proliferation in vivo. The difference between the two tracers is small and the extrapolation from mice to human difficult, and thus it cannot be concluded if one of the tracers would be better than the other for imaging of cancer patients.

The effect of nonsteroidal anti-inflammatory drugs on cAMP-dependent protein kinase-mediated phosphorylation by human chondrocytes in culture.

Although a principal pharmacological action of nonsteroidal anti-inflammatory drugs (NSAIDs) is to blunt eicosanoid synthesis by inhibiting cyclooxygenase, recent evidence indicates that NSAIDs may also interact directly with proteins that control the activity of adenylyl cyclase. Since the only physiological mechanism governing the action of cyclic AMP occurs via activation of its receptor, cyclic AMP-dependent protein kinase (cAMPPk; Kinase A), we determined whether NSAIDs affect intracellular substrate phosphorylation dependent on cAMPPk. The incorporation of 32Pi into cellular proteins that are substrates for cAMPPk was determined in intact human non-arthritic, aged non-arthritic and osteoarthritic chondrocytes in the presence or absence of NSAIDs, namely, sodium meclofenamate, indomethacin, tiaprofenic acid and sodium salicyclate. The transfer of [32P]-ATP was employed to identify phosphoproteins in a membrane fraction prepared from chondrocyte homogenates in the presence or absence of these NSAIDs. The lowest concentration of NSAID was similar to NSAID concentrations achieved during therapy for the arthritides. In intact human chondrocyte strains, activation of cAMPPk by dibutyryl cAMP (dBcAMP) resulted in the phosphorylation of intracellular substrates with an apparent M(r) of 55kD, 42kD, 26kD, 25kD, 22kD, 21.5kD, 20.5kD, and 17kD when examined by autoradiography after 12.5% SDS/PAGE. The NSAIDs augmented or potentiated phosphorylation of these proteins which were cAMPPk-dependent. In the chondrocyte membrane fraction, protein phosphorylation by cAMP was mimicked by isobutylmethylxanthine or by the purified catalytic subunit of bovine cAMPPk. NSAIDs augmented chondrocyte phosphorylation in the chondrocyte membrane fraction.(ABSTRACT TRUNCATED AT 250 WORDS)

Insufficiency fractures of the sacrum. Twenty cases and review of the literature.

Over the past 10 years, 231 insufficiency fractures of the sacrum have been reported in the literature. These fractures, which are due to osteopenia, form a distinct subgroup of pathologic fractures. In 1.8% (n = 20) of the older-than-55-years female patients (total = 1,015) admitted to the authors' Rheumatology department between 1989 and 1991, a fracture of the sacrum was diagnosed. In all cases the diagnosis was confirmed with computed tomography. Frequency, age, sex, diagnosis, underlying diseases, and associated fractures of the 20 cases of insufficiency fractures of the sacrum are described and compared with those previously reported. Insufficiency sacral fracture as a cause of low-back pain in women older than 55 years of age is concluded to be a clinical entity.

123I-iomazenil: a quantitative study of the central benzodiazepine receptor distribution.

Fourteen patients with temporal lobe epilepsy, 9 patients after amygdalohippocampectomy and 3 healthy volunteers were examined with the new benzodiazepine receptor marker 123I-Iomazenil and SPECT. For comparison perfusion SPECT studies with 99mTc-HMPAO were done and a quantitative ROI analysis of the data performed. This quantitative analysis consisted of calculation of right-to-left ratios for 123I-Iomazenil SPECTs, whereby values of 1 were obtained with narrow standard deviations. ROI measurements of the medial occipital, frontal and parietal cortex, the cerebellum and white matter showed a pattern of benzodiazepine receptor concentration in concordance with that previously found in PET and autoradiographic studies, if 123I-Iomazenil ROIs were normalized to the corresponding 99mTc-HMPAO ROIs. The abnormal distribution in the temporal lobes will not be discussed in this paper.

Evaluation of a multicentre study with Iomazenil--a benzodiazepine receptor ligand.

After showing in an earlier publication that Iomazenil is a potent benzodiazepine receptor antagonist, the substance has been distributed to 11 clinical centres in Europe for further tests. The protocol asked for volunteers, epileptic cases and patients with Alzheimer's disease. Prior to the Iomazenil examination, flow images by perfusamine or HMPAO were required, and as comparative methods EEG, computed tomography (CT) and magnetic resonance imaging (MRI) were performed. The results allowed first the determination of the normal distribution of the benzodiazepine receptors in the human brain. The highest uptake was found in medial occipital cortex. Second, the evaluation of the epileptic cases shows a 100% positive prediction value for Iomazenil compared to 92% for flow images. Negative prediction values were calculated as 81% for Iomazenil and 54% for flow images. Furthermore, one group reported the successful diagnosis of Alzheimer's disease at an early stage. The visual image examination was tentatively compared to a more objective semiquantitative one based on quotients of corresponding left/right regions of interest. This semiquantitative method has not proved successful yet, but the problems have been identified. A more precise protocol for further studies is therefore proposed.

Determination of polychlorinated dibenzo-p-dioxins and dibenzo-furans in solid residues from wood combustion by HRGC/HRMS.

PCDD/PCDF were determined in solid samples from wood combustion. The samples included grate ashes, bottom ashes, furnace ashes as well as fly and cyclone ashes. The solid waste samples were classified into bottom and fly ash from native wood and bottom and fly ash from waste wood. For each of the four classes concentration distribution patterns from individual congeners, the sums of PCDD/PCDF and the international toxicity equivalents (I-TEQ) values are given. The I-TEQ levels of fly ash from waste wood burning can be approximately up to two thousand times higher than the values from fly ashes of natural wood. The I-TEQ levels in bottom ashes from waste wood combustion systems are as low as the corresponding ashes from the combustion of native wood. Grate ash samples from waste wood combustion systems with low carbon burnout show high levels of PCDD/PCDF.

Quality assurance and quality control methods for animal and human applications of no-carrier-added 6-[18F]fluorodopa for PET investigations of dopaminergic systems.

No-Carrier-Added (NCA) 6-[18F]fluoro-L-dopa (6-FDOPA) is being produced routinely for PET investigations of dopaminergic systems at our Institute. We describe here in detail the quality assurance methods involved in its multi-step developmental stage as a radiopharmaceutical. A method to remove toxic copper ions to prepare an injectable solution is described. The stability and shelf-life of NCA 6-FDOPA was also examined and results are discussed. Quality control involved three major aspects: (a) chemical purity, (b) radiochemical purity and (c) enantiomeric excess. A method for quick quality control of individual batch preparations is described.

Biological agents in monotherapy for the treatment of rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic, systemic, inflammatory disease, which results in joint destruction and permanent disability. The advent of disease-modifying antirheumatic drugs (DMARDs) has made a profound impact on the outcome and prognosis of RA. Methotrexate (MTX) is a central agent in RA therapy, and is used either alone or in combination with biological DMARDs. However, a large proportion of RA patients (20%-40%) either do not respond to or are unable to tolerate MTX or the alternative agents used in place of MTX (including leflunomide, sulfasalazine, azathioprine, hydroxycholoquine and combination DMARDs). For these patients, monotherapy with biological DMARDs is a key treatment option that balances tolerability with improved clinical outcomes. This article reviews the data for four biological agents approved for use as monotherapy in Switzerland (adalimumab, certolizumab pegol, etanercept and tocilizumab) in order to formulate a consensus statement on their roles in biologic monotherapy of RA.