Near-infrared luminescence and RNA cleavage ability of lanthanide Schiff base complexes derived from N,N'-bis(3-methoxysalicylidene)ethylene-1,2-diamine ligands.

A complete series of lanthanide Schiff base salen-type complexes were prepared with trivalent lanthanide ions (Ln3+) and the N,N'-bis-(3-methoxysalicylidene)ethylene-1,2-diamine ligand (Ln3+=La3+, Ce3+, Pr3+, Nd3+, Sm3+, Eu3+, Gd3+, Tb3+, Dy3+, Ho3+, Er3+, Tm3+, Yb3+, Lu3+). Three unique crystal structures of La3+ and Pr3+N,N'-bis-(3-methoxysalicylidene)ethylene-1,2-diamine complexes, with the La3+ complex prepared in two different synthetic approaches, are reported, namely a dimeric [La(H2L)(NO3)3]2 (H2L=N,N'-bis-(3-methoxysalicylidene)ethylene-1,2-diamine) complex, an asymmetric two-centered [La2(H2L)2(NO3)6] complex and a discrete mononuclear [Pr(H2L)(NO3)2(H2O)2] complex. For Nd3+ and Sm3+, an isotypic mononuclear [Nd(H2L)(NO3)3] and 1D polymeric [Sm(H2L)(NO3)3(MeOH)]n structure was obtained, respectively. The whole series of complexes was tested for their ability to cleave the 20-mer RNA oligonucleotide 5'-AGC-GAU-AAG-AUU-CAU-AUA-UC-3'. Additionally three complexes (Ln3+=Nd3+, Sm3+, Ho3+) were tested for the cleavage of the 12-mer RNA oligonucleotide 5'-GCA-CCC-UGU-CAG-3'. A detailed luminescence study was additionally carried out and revealed that the Eu3+ complex emitted bright red light upon excitation at both 285.8nm and 394.4nm. The Nd3+, Er3+, and Yb3+ complexes showed strong emission in the near-infrared region after excitation at 380nm.

Offline comprehensive liquid chromatography in combination with a Deoxyribonuclease I immobilized enzymatic reactor for selective screening of oligonucleotide mixtures.

The development of a comprehensive ion-pair chromatography-immobilized enzyme reactor×ion-pair chromatography (IPC-IMER×IPC) methodology for the advanced characterization of DNA/RNA oligonucleotides (ONs) mixtures has been carried out. More in detail, a DNase I IMER has been coupled to IPC in the post column configuration, followed by the collection of the eluting fractions and reanalysis by IPC. The effect of the mobile phase over the IMER activity was qualitatively evaluated. The methodology proved to generate relevant ON degradation profiles that might be correlated with the ON stability towards nucleases. Moreover, this platform shows potential for its further implementation in selective analysis of ON mixtures and in mapping studies.

Hyphenation of a Deoxyribonuclease I immobilized enzyme reactor with liquid chromatography for the online stability evaluation of oligonucleotides.

The stability of antisense oligonucleotides (ONs) toward nucleases is a key aspect for their possible implementation as therapeutic agents. Typically, ON stability studies are performed off-line, where the ONs are incubated with nucleases in solution, followed by their analysis. The problematics of off-line processing render the detailed comparison of relative ON stability quite challenging. Therefore, the development of an online platform based on an immobilized enzyme reactor (IMER) coupled to liquid chromatography (LC) was developed as an alternative for improved ON stability testing. More in detail, Deoxyribonuclease I (DNase I) was immobilized on epoxy-silica particles of different pore sizes and packed into a column for the construction of an IMER. Subsequently, the hyphenation of the IMER with ion-pair chromatography (IPC) and ion-exchange chromatography (IEC) was evaluated, leading to the successful development of two online methodologies: IMER-IPC and IMER-IEC. More specifically, natural and modified DNA and RNA oligonucleotides were used for testing the performance of the methodologies. Both methodologies proved to be simple, automatable, fast and highly reproducible for the quantitative and qualitative evaluation of ON degradation. In addition, the extended IMER life time in combination with a more straightforward control of the reaction kinetics substantiate the applicability of the IMER-LC platform for ON stability tests and its implementation in routine and research laboratories.

Combining liquid chromatography with multiplexed capillary gel electrophoresis for offline comprehensive analysis of complex oligonucleotide samples.

Offline two dimensional liquid chromatography (LC)×capillary gel electrophoresis (CGE) and LC×(24) multiplexed-CGE methodologies were developed for the separation of oligonucleotides of therapeutic size. Both ion-pair chromatography (IPC) and ion-exchange chromatography (IEC) were studied as methods for the first dimension and single and multiplexed capillary electrophoresis methods in entangled polymer solutions were used for the second dimension separations. Electrokinetic and pressure injection were evaluated for the analysis of the collected LC fractions. The comprehensive separation was optimized with standard mixtures of poly adenosine, thymidine, cytosine and uracil homodeoxyoligonucleotides up to 35 bases long. Highly orthogonal methodologies and overall peak capacities of 6435 and 6993 for IPC×CGE and IEC×CGE, respectively, were obtained within a few hours analysis time.

Effect of psoriasis activity on metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 in plasma and lesional scales.

The aim of this study was to evaluate the association between psoriasis severity and concentrations of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in plasma and scales from psoriatic lesions, measured with an enzyme immunoassay in 24 patients and analysed with respect to psoriasis area and severity index (PASI). The mean plasma concentrations of both proteins in psoriatic patients significantly exceeded the control values. The proteins were also detectable in scales. There was a significant correlation between plasma MMP-1 concentration and the disease duration. The PASI values showed significant positive correlation with plasma TIMP-1 and significant negative correlation with MMP-1 content in scales. The highest plasma MMP-1 concentration was observed in patients with mild forms whereas the highest plasma TIMP-1 concentrations were demonstrated in severe forms of psoriasis. Our results confirm the role of these proteins in pathogenesis of psoriasis. In severe forms, a decrease in both MMP-1 and TIMP-1 was observed in scales, suggesting their insufficient tissue expression, which can be a crucial element of psoriasis aggravation.

Plasma transforming growth factor beta1 as a biomarker of psoriasis activity and treatment efficacy.

Transforming growth factor-beta(1) (TGFbeta(1)) is thought to be an inhibitor of the keratinocyte hyperproliferation associated with psoriasis. The aim of this study was to evaluate plasma TGFbeta(1) and TGFbeta(2) concentrations in psoriatic patients as possible indicators of treatment efficacy. TGFbeta concentrations were measured in the plasma of 26 patients with psoriasis using an enzyme immunoassay and analysed with respect to the psoriasis area and severity index (PASI) before and after treatment with salicylic acid and/or sulphur followed by dithranol ointment. Baseline plasma concentrations of both TGFbeta(1) and TGFbeta(2) (20.3+/-2.2 ng ml(-1) and 0.14+/-0.02 ng ml(-1), respectively) did not differ significantly from control values (18.3+/-1.6 ng ml(-1) and 0.14+/-0.03 ng ml(-1), respectively). However, a significant positive correlation (r=0.69) between the baseline PASI and TGFbeta(1), but not TGFbeta(2), values was demonstrated. The pretreatment TGFbeta(1) concentration in patients with a PASI >/=15 (26.6+/-3.2 ng ml(-1)) was significantly higher than control values. There were no significant elevation of pretreatment TGFbeta(1) concentrations in patients with a PASI<15, or with respect to TGFbeta(2) in both groups. Treatment caused a significant decrease in TGFbeta(1), but only in patients with a PASI>/=15. Patients with baseline TGFbeta(1) concentrations exceeding the mean of the control group had a PASI value that was significantly higher than that of patients with a TGFbeta(1) concentration below the mean of the controls. These results confirmed an association between plasma TGFbeta(1) concentration and psoriasis severity, and demonstrated its normalization during treatment. Measurement of TGFbeta(1) in plasma should be considered as a possible biomarker of psoriasis activity during its management.

Soluble tumor necrosis factor-alpha receptor type 1 during selenium supplementation in psoriasis patients.

OBJECTIVE: Tumor necrosis factor-alpha (TNF-alpha) and its receptors play important roles in the induction and maintenance of psoriatic lesions. Selenium (Se), a trace element with immunomodulatory properties, is usually decreased in psoriasis patients. We examined the influence of Se supplementation on soluble TNF-alpha receptor type 1 (sTNF-R1) and topical treatment in psoriasis patients. METHODS: The study was conducted in between January and June 2002. Twenty-two inpatients with active plaque psoriasis received topical treatment with 5% salicylic acid ointment, 0.1% to 0.3% dithranol ointment, and 200 microg daily of Se as selenomethionine (SeMet; n = 11, group 1) or placebo (n = 11, group 2) for 4 wk. Psoriasis Area and Severity Index (PASI) score and Se and sTNF-R1 concentrations were assessed at baseline and every 2 wk. Control sera were obtained from 10 healthy subjects. For statistical analysis, parametric tests were used, and the level of significance was set at P = 0.05. RESULTS: The baseline sTNF-R1 levels were 1.87 +/- 0.58 ng/mL (1.98 +/- 0.44 ng/mL in group 1 and 1.75 +/- 0.69 ng/mL in group 2, P = 0.34) in psoriasis patients and 1.65 +/- 0.25 ng/mL in control subjects (P = 0.17); baseline Se concentrations were 48.31 +/- 13.20 microg/L (48.31 +/- 13.20 microg/L in group 1 and 50.35 +/- 13.49 microg/L in group 2, P = 0.41) in psoriasis patients and 58.30 +/- 17.21 microg/L in control subjects (P = 0.05). A positive correlation between PASI and sTNF-R1 was noticed (r = 0.36, P = 0.04; r = 0.51 in group 1 and r = 0.18 in group 2). After 4 wk, almost complete remission of skin lesions was achieved in both groups, but the PASI score was higher in group 1 than in group 2 (4.30 +/- 3.92 and 1.67 +/- 1.17, respectively; P < 0.05). TNF-R1 levels were 1.81 +/- 0.42 ng/mL in group 1 and 1.33 +/- 0.40 ng/mL in group 2 (P = 0.01), and the correlation between PASI score and TNF-R1 level became inverse (r = -0.24 in group 1 and r = -0.59 in group 2). Se concentrations were 107.51 +/- 18.08 microg/L in group 1 and 56.83 +/- 15.32 microg/L in group 2 (P < 0.01). CONCLUSIONS: Increased level of sTNF-R1 may be an indicator of active psoriasis. Supplementation with selenomethionine was ineffective as adjuvant treatment in plaque psoriasis and may contribute to the maintenance of elevated TNF-R1 concentration in psoriasis patients despite the remission of skin lesions.

Association between psoriasis severity and transforming growth factor beta(1) and beta (2) in plasma and scales from psoriatic lesions.

Psoriasis is an inflammatory skin disorder with hyperproliferation of keratinocytes, that can be the result of insufficient inhibitory effect of transforming growth factors-beta (TGF-beta). The aim of this study was to evaluate an association between TGF-beta(1) and -beta(2) in plasma or scales from psoriatic lesions and the severity of the disease. TGF-beta concentrations were measured with an enzyme immunoassay in 41 patients with psoriasis. The mean plasma concentrations of TGF-beta(1) and TGF-beta(2) in patients were: 15.7 +/- 1.4 and 0.15 +/- 0.02 ng/ml respectively. It was also detectable in scales and varied from 24 to 1159 and from 0 to 2.95 pg/mg protein respectively. Plasma TGF-beta(1) correlated significantly with psoriasis area and severity index (PASI). Significant correlation was also demonstrated between TGF-beta(1) concentration in scales and sedimentation rate or the disease duration. There were no correlation between PASI and plasma TGF-beta(2), scales TGF-beta(1) and TGF-beta(2). The highest mean concentration of TGF-beta(1) in scales of patients with mild form of the disease (203 +/- 65 pg/mg protein) and the lowest in severe form (147 +/- 54 pg/mg protein) have been shown. These findings demonstrated association between PASI and plasma levels of TGF-beta(1), that should be considered as a possible indicator of psoriasis activity.

[Spreading of HIV infection and syphilis in Poland in 1988-1998]. / Szerzenie sie zakazenia HIV i kily w Polsce w latach 1988-1998.

OBJECTIVE: To evaluate the epidemiological threat of HIV infection and early syphilis (es) in Poland with regard to the principal modes of HIV transmission and the relationship between HIV and es in 1988-98. METHODS: We analyzed es incidence rates and newly notified HIV cases and their trends in four regions: East (E), South (S), West (W) and Central (C); proportion of injecting drug users (IDUs), men who have sex with men (MSM), and persons of unknown mode of HIV transmission (UMTPs) among reported HIV infections and correlation between HIV infection and es incidence rates. RESULTS: HIV infection notification rate curves in four regions were two-phase: since 1988 they were parabolic with the highest values in 1989-90 and the lowest in 1993-95, then they had increasing trends in four regions. Proportion of MSM was the highest (> 10% among HIV+) in C till 1996, since 1993 it increased in E, W and S. Proportion of IDUs was 80% in late 80s, then decreased (50-60% in 1993) and rose in E, S and C since 1993. Proportion of UMHTs was inversely correlated to the percentage of IDUs in E, W and C and increased in W and C since 1993. In the analysed period es incidence rate slightly increased only in E and positively correlated to HIV incidence in this region. CONCLUSIONS: IDUs in Poland may constitute a 'core group' contributing to the spread of HIV together with other sexually transmitted infections from former USSR. Increasing number of UMTPs makes the epidemiological situation of HIV infection difficult to evaluate.