B Cell Synovitis and Clinical Phenotypes in Rheumatoid Arthritis: Relationship to Disease Stages and Drug Exposure.

OBJECTIVE: To define the relationship of synovial B cells to clinical phenotypes at different stages of disease evolution and drug exposure in rheumatoid arthritis (RA). METHODS: Synovial biopsy specimens and demographic and clinical data were collected from 2 RA cohorts (n = 329), one of patients with untreated early RA (n = 165) and one of patients with established RA with an inadequate response to tumor necrosis factor inhibitors (TNFi-IR; n = 164). Synovial tissue was subjected to hematoxylin and eosin and immunohistochemical staining and semiquantitative assessment for the degree of synovitis (on a scale of 0-9) and of CD20+ B cell infiltrate (on a scale of 0-4). B cell scores were validated by digital image analysis and B cell lineage-specific transcript analysis (RNA-Seq) in the early RA (n = 91) and TNFi-IR (n = 127) cohorts. Semiquantitative CD20 scores were used to classify patients as B cell rich (≥2) or B cell poor (<2). RESULTS: Semiquantitative B cell scores correlated with digital image analysis quantitative measurements and B cell lineage-specific transcripts. B cell-rich synovitis was present in 35% of patients in the early RA cohort and 47.7% of patients in the TNFi-IR cohort (P = 0.025). B cell-rich patients showed higher levels of disease activity and seropositivity for rheumatoid factor and anti-citrullinated protein antibody in early RA but not in established RA, while significantly higher histologic synovitis scores in B cell-rich patients were demonstrated in both cohorts. CONCLUSION: We describe a robust semiquantitative histologic B cell score that closely replicates the quantification of B cells by digital or molecular analyses. Our findings indicate an ongoing B cell-rich synovitis, which does not seem to be captured by standard clinimetric assessment, in a larger proportion of patients with established RA than early RA.

Enzyme assays using sensor arrays based on ion-selective carbon nanotube field-effect transistors.

In the fields of clinical diagnostics and point-of-care diagnosis as well as food and environmental monitoring there is a high demand for reliable high-throughput, rapid and highly sensitive assays for a simultaneous detection of several analytes in complex and low-volume samples. Sensor platforms based on solution-processable electrolyte-gated carbon nanotube field-effect transistors (CNT-FETs) are a simple and cost-effective alternative for conventional assays. In this work we demonstrate a selective as well as direct detection of the products of an enzyme-substrate interaction, here the for metabolic processes important urea-urease system, with sensors based on spray-coated CNT-FETs. The selective and direct detection is achieved by immobilizing the enzyme urease via certain surface functionalization techniques on the sensor surface and further modifying the active interfaces with polymeric ion-selective membranes as well as pH-sensitive layers. Thereby, we can avoid the generally applied approach for a field-effect based detection of enzyme reactions via detecting changes in the pH value due to an on-going enzymatic reaction and directly detect selectively the products of the enzymatic conversion. Thus, we can realize a buffering-capacity independent monitoring of changes in the substrate concentration.

Selective ion-sensing with membrane-functionalized electrolyte-gated carbon nanotube field-effect transistors.

In this work the ion-selective response of an electrolyte-gated carbon-nanotube field-effect transistor (CNT-FET) towards K(+), Ca(2+) and Cl(-) in the biologically relevant concentration range from 10(-1) M to 10(-6) M is demonstrated. The ion-selective response is achieved by modifying the gate-electrode of an electrolyte-gated CNT-FET with ion-selective membranes, which are selective towards the respective target analyte ions. The selectivity, assured by the ion-selective poly(vinyl chloride) based membrane, allows the successful application of the herein proposed K(+)-selective CNT-FET to detect changes in the K(+) activity in the µM range even in solutions containing different ionic backgrounds. The sensing mechanism relies on a superposition of both an ion-sensitive response of the CNT-network as well as a change of the effective gate potential present at the semiconducting channel due to a selective and ion activity-dependent response of the membrane towards different types of ions. Moreover, the combination of a CNT-FET as a transducing element gated with an ion-selective coated-wire electrode offers the possibility to miniaturize the already well-established conventional ion-selective electrode setup. This approach represents a valuable strategy for the realization of portable, multi-purpose and low-cost biosensing devices.

Side population/ABCG2-positive cells represent a heterogeneous group of haemopoietic cells: implications for the use of adult stem cells in transplantation and plasticity protocols.

Murine side population (SP) cells may have an increased ability to engraft lethally irradiated mice and lack CD34 expression. Strategies using CD34 as a primary marker of haemopoietic stem cells may therefore result in the exclusion of a primitive stem cell population. The molecular basis for the murine SP phenotype has been attributed to the multidrug-resistance transporter ABCG2. This study aimed to investigate ABCG2 expression from a variety of human sources and investigate the relationship between ABCG2 expression, the SP phenotype, and expression of markers such as CD34 and CD133. SP cells were observed in different haemopoietic sources, but a significant increase in the number of SP cells was observed in PB following granulocyte colony-stimulating factor mobilisation. No direct correlation between the frequency of SP cells and the expression of ABCG2 was observed. SP cells were identified in both lineage-positive and lineage-negative population and ABCG2 expression was enriched in lineage-negative SP cells. Lineage-negative SP cells were devoid of CD34 expression but enriched for CD133. Subsequent analysis revealed that ABCG2 and CD133 are coexpressed. Together, these data suggest that the ABCG2 transporter is neither required nor responsible for the SP phenotpye in many human blood cells.

CCR1 chemokine receptor expression isolates erythroid from granulocyte-macrophage progenitors.

Simple methods that separate progenitor cells of different hemopoietic lineages would facilitate studies on lineage commitment and differentiation. We used an antibody specific for the chemokine receptor CCR1 to examine mononuclear cells isolated from cord blood samples. When CD34(+) cells were separated into CD34(+)CCR1(+) and CD34(+)CCR1(-) cells and plated in colony-forming assays, the granulocyte/macrophage progenitors were found almost exclusively in the CD34(+)CCR1(+) cells. In contrast, the CD34(+)CCR1(-) cells contained the majority of the erythroid progenitors. There was a highly significant difference (P<0.002) in the total percentage distribution of both granulocyte-macrophage colony-forming cells and erythroid burst-forming units between the two populations. This is the first report of separation of erythroid progenitors from granulocyte/macrophage progenitors using a chemokine receptor antibody in cord blood samples. These results suggest that at the clonogenic progenitor cell stage the expression of CCR1 might be lineage-specific. This method should prove useful for studies on erythroid progenitor and granulocyte/macrophage differentiation.

The Gas-Phase Electric Dipole Moments of the Symmetric Top Tertiary Butyl Molecules (t)BuX, X=F, Cl, Br, I, CN, and NC.

The electric dipole moments of symmetric top tertiary butyl derivatives were determined from Stark effect measurements made with cavity Fourier transform microwave spectroscopy under conditions of supersonic expansion. The resulting values are 1.9562(15), 2.1817(16), 2.2574(17) and 2.2122(17) D for tertiary butyl fluoride, chloride, bromide, and iodide, and 4.0129(30), and 4.0640(31) for tertiary butyl cyanide and isocyanide, respectively. The experimental values are compared with ab initio calculations and with experimental and calculated dipole moments for the corresponding methyl derivatives. Copyright 2001 Academic Press.

[Ultrasonographic diagnosis of fetal congenital abnormalities]. / Diagnostyka ultrasonograficzna wad wrodzonych u plodu.

Retrospective analysis of ultrasonographic examinations in 81 women who delivered newborns with congenital malformations was performed. Totally 147 anatomical defects in 83 newborns were found. 85 of 147 abnormalities (57.8%) were detected prenatally, most commonly (79.6%) in the third-trimester. All cases of coverings defects, as well as, all cases of anencephaly were diagnosed with ultrasonography. Malformations of urinary tract in 94.4%, central nervous system in 82.9%, osteoarticular system in 30%, heart defects in 27.8% and face abnormalities in 21.4% were already sonographically visualized before birth.

The angleICI Bending Satellites in the Millimeter-Wave Rotational Spectra of CH(2)I(2) and CD(2)I(2).

Rotational transitions in the first four excited states of the low-frequency angleICI bending mode, nu(4), have been assigned in the mm-wave rotational spectra of CH(2)I(2) and of CD(2)I(2). Measurements of transition frequencies, made over the frequency region 167-326 GHz and for J" up to 190, allowed determination of sextic level spectroscopic constants for all states. The changes in spectroscopic constants with vibrational excitation show very small anharmonicity, in spite of the very low frequency of this mode (121 cm(-1)). Vibrational excitation affects the moments of inertia in such a way that the planar moment P(b), about the plane perpendicular to both angleICI and angleHCH, is practically invariant. Vibrational change in P(c), the moment along the principal axis in the HCH plane and perpendicular to the angleHCH bisector, has been successfully reproduced with an ab initio harmonic force field so that there is no discernible vibrational change in angleHCH on excitation of angleICI. Finally, the change in P(a) leads to estimated vibrational change of +0.12 degrees in the value of angleICI itself. Copyright 2000 Academic Press.

The Rotational Spectrum of CBrClF2 (Halon BCF): II. The Lowest Excited Vibrational States and Nuclear Quadrupole Coupling Tensors

The analysis of the millimeter-wave rotational spectrum of the well-known halon molecule CBrClF2 was extended to rotational transitions in the excited vibrational states v9 = 1 (307 cm-1) and v5 = 1 (337 cm-1). The two states are appreciably coupled by a- and b-axis Coriolis interaction which was treated explicitly and spectroscopic constants were determined from measurements at frequencies from 121 to 330 GHz and J" from 18 to 111. The present work, together with our previous study (1996. J. Mol. Spectrosc. 177, 240-250), results in accurate constants in the rotational Hamiltonian for the four lowest vibrational states of C79Br35ClF2 and C81Br35ClF2. The double nucleus hyperfine structure in low-J transitions of the two 35Cl isotopomers of CBrClF2 was also measured in the region 3-14 GHz with a supersonic beam, cavity Fourier transform microwave spectrometer. All constants in both the inertial and the principal nuclear quadrupole coupling tensors have been determined for both Br and Cl nuclei. Copyright 1997 Academic Press. Copyright 1997Academic Press

Catecholamine and blood pressure regulation by gonadotropin-releasing hormone analogs in amphibians.

Analogs of gonadotropin-releasing hormone (GnRH) occur in the brain, plasma, and sympathoadrenal system of anuran amphibians. The present experiments studied the effects of GnRH and [Trp7, Leu8]-GnRH on plasma catecholamines and cardiovascular function in conscious adult bullfrogs (Rana catesbeiana) and cane toads (Bufo marinus). Both GnRH analogs elicited dose-dependent (0.1-1 nmol.kg-1) increases in arterial norepinephrine, epinephrine, and blood pressure levels when injected intravenously into toads. In bullfrogs, [Trp7, Leu8]-GnRH (1 nmol.kg-1) increased arterial norepinephrine concentration approximately 10-fold without affecting the concentrations of norepinephrine sulfate, norepinephrine glucuronide, epinephrine, epinephrine sulfate, or epinephrine glucuronide. The noradrenergic response of bullfrogs to [Trp7, Leu8]-GnRH was specific to the neurohormone because it could be inhibited by [D-pGlu1, D-Phe2, D-Trp3,6]-GnRH. The sympathomimetic activities of the GnRH analogs did not depend on changes in temperature, which occur seasonally in natural habitats, because similar noradrenergic responses were observed at 4 and 22 degrees C. GnRH and [Trp7, Leu8]-GnRH (0.01-10 nmol.kg-1) did not raise arterial blood pressure in bullfrogs despite their pressor actions in toads. This interspecific difference was remarkable because cardiovascular responses to norepinephrine, angiotensin II, and vasotocin in bullfrogs were similar to those in toads. The parallels between catecholamine and blood pressure responses suggest that epinephrine is the principal mediator of the blood pressure response to native GnRH analogs in toads. In bullfrogs, [Trp7, Leu8]-GnRH mobilizes norepinephrine but not epinephrine, and the noradrenergic effect is insufficient to raise blood pressure. These observations are consistent with a physiological role for native GnRH analogs in the regulation of the sympathoadrenal system in anuran amphibians.

IgG and IgE antibody response following Pollinex-R immunotherapy.

Specific antibodies to ragweed pollen antigens were studied in 22 patients given preseason Pollinex-R (glutaraldehyde-modified ragweed tyrosine-adsorbate) (PR), 17 without immunotherapy, and 8 on maintenance doses of aqueous ragweed extracts. The PR-group showed about fourfold increases in IgG antibodies in season when compared with pretreatment levels (P less than .001). IgG antibody changes from before season to season in other groups and IgE antibody changes in all three groups were not significant. Despite this, IgG antibodies in season had fallen from their peak achieved with PR. About one-third of PR-treated patients still manifested significant symptomatology irrespective of changes in IgG antibodies. We thus conclude that although changes in specific IgG may be an important correlate in PR-immunotherapy, additional mechanisms for clinical responsiveness remain to be clarified.

[Detection of pentachlorophenol in natural rubber latex]. / Wykrywanie pieciochlorofenolu w lateksie kauczuku naturalnego

The method of detection of pentachlorophenol in natural rubber latex is proposed. Pentachlorophenol is isolated from other nonrubber-like substances by thin-layer chromatography and identified by spectroscopic method in UV-light. Isolation of pentachlorophenol is carried out from water extracts obtained from the dry caoutchouc films, so the same method can be used for examination of the rubber articles designed for the medicinetoo.