The role of occlusion and micro-incontinence in the pathogenesis of penile lichen sclerosus: an observational study of pro-inflammatory cytokines' gene expression.

PURPOSE: To assess the expression of selected cytokines in penile lichen sclerosus (PLS) and associate them with the occurrence of micro-incontinence (MI) in different stages of PLS. METHODS: The skin biopsies from 49 PLS affected, and 13 from nonlesional foreskins (healthy control adult males undergoing circumcision due to phimosis caused by short frenulum) were obtained. All specimens were used for RNA extraction and RT-qPCR. Quantitative assessment of the gene expression of interleukin 1-A (IL-1A), interleukin 1-B (IL-1B), interleukin 1 receptor antagonist (IL-1RN), interleukin 6 (IL-6), transforming growth factor ß1 (TGF-ß1), and interferon-gamma (INF-γ) was performed. To determinate the presence of MI, the patients were asked about voiding patterns, especially leaking tiny drops of urine from the urethral meatus after urination. RESULTS: IL-1A, IL-6, and INF-γ mRNA levels were approximately 150, 16, and 59 times higher in PLS than in control samples, respectively. The highest IL-1A mRNA levels were observed in early PLS (n = 13), INF-γ in moderate PLS (n = 32), while IL-6 in severe PLS (n = 4). MI was noted in 45 PLS patients vs. 0 in control (p < 0.0001). IL-1A and IL-6 vs control ratios were concentration (ca.) 400 and 30 times higher, respectively, in MI PLS samples than in PLS without MI. CONCLUSION: Occlusion and irritating urine effect are associated with the clinical progression of penile LS with increased mRNA expression of IL-1A, INF-γ, and IL-6 pro-inflammatory cytokines in the foreskin.

Induction of hormone-sensitive lipase/cholesteryl esterase gene expression by C/EBPα independently of the PKA pathway in the adrenocortical Y-1 cells.

The effect of C/EBPα on the expression of LIPE gene encoding hormone-sensitive lipase/cholesteryl esterase (HSL) was investigated in Y-1 CCL79 cells. It was found that transfection of these cells with the vector overexpressing C/EBPα increased both the level of LIPE transcript, measured by RT-qPCR, and the luminesce emitted by luciferase reporter gene fused to the -2150 fragment of LIPE promoter. Activation of adenylyl cyclase by forskolin resulted in 2.5-fold increase in the intensity of luminescence and over 3-fold increase in luminescence was observed when the cells were cotransfected with the vector overexpressing C/EBP. The incubation of C/EBP-cotransfected cells with forskolin caused over 6-fold increase in the intensity of luminescence, suggesting that the effects of C/EBPα and forskolin are additive. The analysis of sequence of the proximal LIPE promoter showed multiple binding sites for various transcription factors including C/EBPα site, which is located between nucleotides -46 bp and -59 bp. When the Y-1 cells were transfected with the recombinant vector containing -60 bp fragment of LIPE promoter fused to the luciferase reporter gene and were cotransfected with the vector overexpressing C/EBPα, the luminescence increases about 9-fold indicating that C/EBPα stimulates the expression of LIPE by reacting with its response element. The results indicate that C/EBPα stimulates the expression of LIPE independently of the PKA pathway by binding to a response element situated within the -60 bp fragment of LIPE promoter. This suggests that C/EBPα might be involved in the regulation of LIPE expression and thus cholesterol supply for steroid hormone synthesis.

Interatomic potential parameters of CdHe van der Waals complex derived from excitation spectrum of the C1 1(51P1)<--X10+(51S0) vibrational transition.

The first time observed excitation spectrum of the C(1)1(5(1)P(1))<--X(1)0+(5(1)S(0)) transition in CdHe van der Waals molecules is reported. Vibrational spectrum in the UV region (2286.0-2296 A) was excited in a continuous molecular-jet-expansion beam of CdHe seeded in helium using an in-house-built nitrogen-dye laser system. The excitation spectrum exhibits two vibrational components (v'<--v''=0) highly broadened by means of unresolved rotational structure and some additional contributions of "hot-bands" components (v'<--v''=1). The last effect is due to an extremely small separation of the vibrational levels in the ground X(1)0+ state of the CdHe molecule, where v'=0 vibrational level is separated from v''=0 by merely 6.0 cm(-1). It follows therefore that even in an extremely cold environment (T(v) approximately 10K) of a jet-expansion beam the population of v''=1 level is feasible, due to some residual collisions, and hence the v'<--v'=1 transitions are highly probable. The assignment of vibrational bands and numerical analysis of the spectrum was based and obtained with the aid of a rigorous computer simulation of the C(1)1<--X(1)0+ transition including the impact of rotational structure and hot-bands contributions. As a result we obtained optical potential parameters of the C(1)1(5(1)P(1)) state of CdHe molecule which are further discussed in terms of our recent (and only existing) experimental results regarding the X(1)0+, B1(5(3)P(1)) and A0+(5(3)P(1)) states of CdHe as well as in terms of ab initio calculations results.

Smokers with airway obstruction are more likely to quit smoking.

BACKGROUND: Chronic obstructive pulmonary disease (COPD), usually caused by tobacco smoking, is one of the leading causes of morbidity and mortality. Smoking cessation at an early stage of the disease usually stops further progression. A study was undertaken to determine if diagnosis of airway obstruction was associated with subsequent success in smoking cessation, as advised by a physician. METHODS: 4494 current smokers (57.4% men) with a history of at least 10 pack-years of smoking were recruited from 100 000 subjects screened by spirometric testing for signs of airway obstruction. At the time of screening all received simple smoking cessation advice. 1177 (26.2%) subjects had airway obstruction and were told that they had COPD and that smoking cessation would halt rapid progression of their lung disease. No pharmacological treatment was proposed. After 1 year all subjects were invited for a follow up visit. Smoking status was assessed by history and validated by exhaled carbon monoxide level. RESULTS: Nearly 70% attended a follow up visit (n = 3077): 61% were men, mean (SD) age was 52 (10) years, mean (SD) tobacco exposure 30 (17) pack-years, and 33.3% had airway obstruction during the baseline examination. The validated smoking cessation rate in those with airway obstruction was 16.3% compared with 12.0% in those with normal spirometric parameters (p = 0.0003). After correction for age, sex, nicotine dependence, number of cigarettes smoked daily, and lung function, success in smoking cessation was predicted by lower lung function, lower nicotine dependence, and lower tobacco exposure. CONCLUSIONS: Simple smoking cessation advice combined with spirometric testing resulted in good 1 year cessation rates, especially in subjects with airway obstruction.

The structure of the lowest electronic Rydberg state of CdAr complex determined by laser double resonance method in a supersonic jet-expansion beam.

The lowest E1(3Sigma(+)) Rydberg state of the CdAr van der Waals (vdW) complex was investigated by means of an optical-optical double resonance (OODR) method of laser spectroscopy in conjunction with a free jet-expansion molecular beam. Two dye lasers were employed for the two-step excitation. The A0(+)(3Pi(+)) and B1(3Sigma(+)) states were used as intermediates in the excitation process from the X0(+)(1Sigma(+)) ground state. Two types of bound-bound excitation spectra of the E1<--A0(+) and E1<--B1 transitions were recorded indicating the existence of two, well defined minima in the E1-state potential energy (PE) curve. First, considerably deep, with the well depth of D(e)'(E1(2))=1309.0 cm(-1) and second, separated by a positive PE barrier, with D(e)'(E1(2))=24.2 cm(-1). Combination of bound-bound and first-time observed bound-free excitation spectra enabled a complete determination of the spectroscopical parameters of the PE curve of the E1-Rydberg state, the height of the PE barrier and its approximate location. In the excitation spectra of the E1<--B1 transition, a nodal structure of the bound-free transitions was observed and elucidated by a projection of the B1-state vibrational wave-functions onto the E1-state potential barrier and/or onto the repulsive part of the E1-state PE curve. The experimental results of our investigation coincides well with recently published results of ab initio calculation of Czuchaj and co-workers [Chem. Phys. 248 (1999) 1; Chem Phys. 263 (2001) 7; Theor. Chem Acc. 105 (2001) 219].

Laser spectroscopy of CdKr molecules in ultraviolet region.

Excitation spectra of the A0+(5(3)P1)<--X0+(5(1)S0), B1(5(3)P1)<--X0+ and D1(5(1)P1)<--X0+ transitions as well as a fluorescence spectrum of the D1(v') = 16 --> X0+ transition in CdKr van der Waals molecule has been recorded in an experiment of a continuous supersonic molecular beam crossed with a pulsed dye laser beam. A rigorous analysis of the excitation spectra based on Birge-Sponer and LeRoy-Bernstein methods aided with a computer calculation, utilizing a program of near dissociation expansion lead to the X0+- and improved A0+-, B1- and D1-state characterizations. Evidence of a theoretically predicted potential barrier within the B1-state well was found and discussed with regard to the latest result of ab initio calculation. An analysis of bound-bound and bound-free parts of the fluorescence spectrum confirmed a v'-assignment and indicated that Morse and Maitland-Smith M-S (n0, n1) functions are adequate representations of the bound well and repulsive wall of the X0+-state potential energy curve, respectively. All spectroscopic characteristics obtained in this work are compared with newest theoretical studies of this complex.

Double-well potential energy curve of cadmium-krypton molecule in the B1(5(3)P1) excited state.

The real shape of a double-well B1(5(3)P1)-state potential in CdKr van der Waals molecule was reconstructed applying both the experimental data, using a Birge-Sponer method-based analysis of the B1 <-- X0+(5(1)S0) transition in excitation spectrum, and theoretical result of recent ab initio calculation. An inverse perturbation approach method was used for defining an accurate B1-state potential energy curve.

Laser excitation spectrum and spectroscopic potential parameters of Cd2 molecule in the 1u(5(3)P2) energy state.

Excitation spectrum of Cd2 van der Waals complex was observed in a supersonic free-jet expansion beam. Cadmium dimers were seeded in argon environment (a carrier gas) while a pulsed dye laser served as an excitation light source. Well resolved structures of vibrational bands arising from transition between 1u(3pi(u)) and X0g+ (1sigmag+) states were observed. Using a Birge-Sponer method for analyzing the vibrational excitation spectrum, the first-time determination of the spectroscopical potential parameters of the 1u molecular state was performed. We obtained D'e = 723 +/- 10 cm(-1), omega'e = 28.9 +/- 1.0 cm(-1), omega'e x'e = 0.260 +/- 0.002 cm(-1) and R'e = 3.93 +/- 0.05 A. The latter value was estimated with the help of the computer-simulation of the spectrum. The results are compared with recent results of ab initio calculation of Czuchaj et al. (Chem. Phys. Lett. 225 (1994) 233) and demonstrate a reasonable agreement.

Treatment of intestinal Behçet's syndrome with chimeric tumour necrosis factor alpha antibody.

Few patients with Behçet's syndrome have gastrointestinal ulceration. Such patients are difficult to treat and have a higher mortality. Faced with refractory symptoms in two patients with intestinal Behçet's, we used the tumour necrosis factor alpha (TNF-alpha) monoclonal antibody infliximab to induce remission. Both women (one aged 27 years, the other 30 years) presented with orogenital ulceration, pustular rash, abdominal pain, bloody diarrhoea due to colonic ulceration, weight loss, and synovitis. One had thrombophlebitis, digital vasculitis, perianal fistula, and paracolic abscess; the other had conjunctivitis and an ulcer in the natal cleft. Treatment with prednisolone, methyl prednisolone, and thalidomide in one and prednisolone, colchicine, and cyclosporin in the other was ineffective. After full discussion, infliximab (3 mg/kg, dose reduced because of recent sepsis in one, and 5 mg/kg in the other) was administered. Within 10 days the ulcers healed, with resolution of bloody diarrhoea and all extraintestinal manifestations. A second infusion of infliximab was necessary eight weeks later in one case, followed by sustained (>15 months) remission on low dose thalidomide. Remission was initially sustained for 12 months in the other but thalidomide had to be stopped due to intolerance, and a good response to retreatment lasted only 12 weeks without immunosuppression, before a third infusion. The cause of Behçet's syndrome is unknown but peripheral blood CD45 gammadelta T cells in Behçet's produce >50-fold more TNF-alpha than controls when stimulated with phorbol myristate acetate and anti-CD3. Infliximab could have a role for inducing remission in Behçet's syndrome.

Interatomic potentials of cadmium-argon B1(3sigma+) and X0+(1sigma+) states based on near-dissociation expansion and 'hot' bands observed in the B1 <-- X0+ excitation spectrum.

B1(3sigma+) <-- X0+(1sigma+) excitation spectrum of the cadmium-argon van der Waals molecule has been recorded in the experiment of a continuous supersonic molecular beam crossed with a pulsed dye laser beam. The B1-state dissociation energy was directly observed and a long-range behavior of the B1-state intermolecular potential was derived using a near-dissociation expansion procedure of LeRoy and Bernstein. A first-time direct determination of the X0+ ground state characteristics based on 'hot' bands observed is presented as well. The new result is compared with other experimental and theoretical reports available in the literature.

Generation and characterization of an anti-idiotypic antibody specific for intercellular adhesion molecule-1.

Intercellular adhesion molecule-1 (ICAM-1) is an inducible ligand for the LFA-1/MAC-1 family of leukocyte adhesion molecules. The results reported herein show that a mouse monoclonal anti-idiotypic antibody, CA3, specific for R6.5 anti-ICAM-1 mAb, shares conformational homology with the original epitope bound by R6.5. The CA3 bound specifically to R6.5 F(ab) fragments and blocked the binding of R6.5 to its ICAM-1 epitope; no CA3 binding was detected to a second anti-ICAM-1 mAb F(ab) fragment nor to control mouse IgG F(ab) fragments. Similarly, the interaction of CA3 with R6.5 was inhibited by sICAM-1. However, CA3 was ineffective in inhibiting CD18-dependent cell aggregation. A rabbit anti-CA3 response (Ab3) further indicated that CA3 is of the Ab2 beta type. Significant binding of anti-CA3 to sICAM-1 was demonstrated and anti-CA3 competed with R6.5 for binding to sICAM-1. Anti-CA3 bound to both soluble and cell-surface-associated ICAM-1. However, unlike R6.5, significant binding both to reduced sICAM-1 and to native sICAM-1 was exhibited by anti-CA3, whereas the binding of R6.5 to reduced sICAM-1 was undetectable.

Wavelength dependence of fluorescence spectra of liquid coumarin solutions.

The excitation wavelength dependence of steady-state fluorescence spectra of low-viscosity dye solutions is reexamined. Measurements are made on some coumarins in methyl alcohol at room temperature. It is found that the fluorescence spectra depend on the excitation energy and their profile changes are due to the excitation wavelength dependence of reorientation processes in the excited state.

A form of circulating ICAM-1 in human serum.

A circulating form of the usually membrane-bound intercellular adhesion molecule-1 (ICAM-1) was identified and characterized in normal human serum, and in sera from patients with leukocyte adhesion deficiency (LAD). The molecule, designated circulating ICAM-1 (cICAM-1) was detected and quantitated by sandwich ELISA. Levels of cICAM-1 in sera from normal individuals ranged from 100 to 200 ng/ml. Sera from LAD patients had elevated cICAM-1 levels ranging from 200 to 700 ng/ml. The elevated levels of cICAM-1 in LAD sera may be due to an inability to adsorb cICAM-1 to cell-bound LFA-1 or may be an indirect result of the pathology accompanying the syndrome. cICAM-1 bound to mAb specific for four distinct ICAM-1 epitopes localized in domains D1, D2, D4, and D5, and displayed similar molecular size properties as recombinant soluble ICAM-1 on FPLC size-exclusion chromatography. When immobilized via a domain D5-specific mAb, cICAM-1 mediated function (LFA-1)-dependent lymphocyte adhesion equivalent to sICAM-1. These data indicate that cICAM-1 contains most, if not all, of the five extracellular domains of membrane ICAM-1, as well as the ability to bind specifically to LFA-1. The cellular source of cICAM-1 appeared to be from mononuclear cells; only lymphoid cell lines or primary PBMC cultures had detectable levels of cICAM-1 in cell culture supernatants. Because cICAM-1 retains the ability to bind specifically to LFA-1, it may act to regulate cell adhesion by promoting de-adhesion. Alternatively, cICAM-1 may be the indirect consequence of inflammation or tissue damage. As such, the detection of cICAM-1 could be useful as a marker of inflammatory disease.