Human trophoblast cells express the immunomodulator progesterone-induced blocking factor.

Progesterone-induced blocking factor (PIBF) is an immunomoduatory factor with anti-abortive properties. In this study, we present evidence that PIBF is synthesized in the human placenta and determine its cellular source. Expression of PIBF was analysed with polyclonal rabbit anti-human PIBF antibodies against recombinant N-terminal 48kDa PIBF in first trimester and term placental tissues and in the choriocarcinoma cell line JAR by means of immunohistochemistry, confocal laser scanning microscopy of double immunofluorescence labelling, and Western blotting; RT-PCR was performed for analysis of PIBF mRNA in isolated trophoblast cells. PIBF protein is present in human first trimester and term placenta. Double immunofluorescence labelling localised PIBF to the extravillous cytotrophoblast. PIBF is also expressed heterogeneously by syncytiotrophoblast and part of the villous cytotrophoblast. Full-length PIBF mRNA encoded by exons 1-18 is present in isolated first trimester and term villous trophoblast and in the choriocarcinoma cell line JAR. The corresponding 90kDa protein is expressed by JAR cells, first trimester and term villous trophoblast cells. In addition, these cells express PIBF proteins of 50 and 34kDa. Trophoblast is a source of PIBF; its tissue distribution suggests a role both in systemic and local (decidual) immunoregulation.

Development and validation of a liquid chromatographic method for the determination of hydroxymethylfurfural and alpha-ketoglutaric acid in human plasma.

Hydroxymethylfurfural (HMF) and alpha-ketoglutaric acid (KG) have been recently investigated as potential cancer cell damaging agents. We herein report for the first time a validated quantitative assay for their simultaneous determination in human plasma which is amenable to be applied in the future screening of the target compounds in human probands in order to properly design a targeted chemotherapeutic regimen for certain types of malignant tumors. A simple liquid chromatographic method in conjunction to derivatization after a two-step optimized solid phase clean-up procedure is described. The method is based on the reaction of HMF and KG with 2-nitrophenylhydrazine or 2,4-dinitrophenylhydrazine in an aqueous environment. Reaction conditions were studied with respect to pH, reagent volume, reaction temperature and time. Exact testing of such parameters beside careful selection of the mobile phase composition rendered feasible the quantification of the chemically significantly differing analytes along a single chromatographic run. The formed derivatives could be separated isocratically by reversed-phase LC on a C(8)-column. Detection in the UV and in the visible range is possible. Results showed good recovery and reproducibility with detection limits (S/N=3) down to 2 picomoles analyte on column. Resolution of the syn and anti geometric isomers of the HMF and KG derivatives is possible. The isomeric ratio in relation to the reaction pH is discussed.

Hydroxymethylfurfural: an enemy or a friendly xenobiotic? A bioanalytical approach.

Hydroxymethylfurfural (HMF), a well-known heterocyclic Maillard reaction product, has often been studied for its potential toxic, mutagenic, and carcinogenic effects. Recent clinical studies, however, have strongly suggested that HMF might have exciting antitumor potential. We report on the development and validation of a bioanalytical assay for HMF that could be suitable as a basis for pharmacokinetic models in cancer patients. Two strategies were tested, i.e., direct and indirect methodologies. A direct isocratic LC determination at 283 nm was designed. Two indirect attempts involved derivatization coupled to HPLC-UV. It was possible to resolve the stereoisomers of the HMF derivative, and factors influencing their equilibrium ratio are discussed. HMF was extracted from the biomatrix by solid-phase extraction using different cartridges. A comparative study was made of the implemented methods as well as the extraction protocols. Both indirect assays proved to be more sensitive and were used to assess HMF quantitatively in human plasma. However, the newly introduced derivatization conditions led to the highest sensitivity with a LOD (S/N ratio = 3) of at least 2 pmol analyte on column. The assay selectivity was satisfactory in pre- and post-dose real samples. The mean recoveries of the assays were 79% and 89%, with acceptable accuracies and reproducibilities. Figure Schematic representation of hydroxymethylfurfural (HMF) in human plasma.

Determination of phylloquinone and cholecalciferol encapsulated in granulates formed by melt extrusion.

Vitamin K1 (phylloquinone) and vitamin D3 (cholecalciferol) play a dominant role in bone metabolism. Both vitamins are sensitive to ultraviolet radiation, oxygen and other environmental influences. For this reason a special extrusion technology was developed, that enables an encapsulation of these sensitive substances in a matrix of carbohydrates and hydrogenated carbohydrates. To exclude decomposition products possibly originating under process conditions quantitative analysis was carried out by HPLC/UV using a modified method based on United States Pharmacopoeia. Under the used chromatographic conditions it has to be possible to separate cis-phylloquinone, trans-phylloquinone and phylloquinone 2,3-oxide, as well as pre-cholecalciferol, cis-cholecalciferol and trans-cholecalciferol. A silica column as stationary phase and a mixture of n-hexane and 1-amyl alcohol as mobile phase were used for quantification. UV detection ensued at 254 nm. A linear relationship between peak area and concentration was found over almost two orders of magnitude for cis-phylloquinone, trans-phylloquinone and cholecalciferol. The detection limits (S/N 3) on column were 0.1 microg for phylloquinone and 0.4 microg for cholecalciferol. Analytical results showed that the vitamins were encapsulated sufficiently in the used carbohydrate matrix and that they were protected against environmental influences. After granulation process all of the samples tested met the pharmacopoeial requirements.

[Peripheral analgesic effect of intra-articularly applied clonidine]. / Periphere analgetische Wirkung durch intraartikulär verabreichtes Clonidin.

BACKGROUND AND AIM: Clonidine applied intra-articularly into the knee joint has a peripheral analgesic effect. We examined intra-articularly injected clonidine to determine whether resorption with a measurable systemic concentration could be detected. METHODS: A randomised, placebo-controlled double-blind study was carried out on patients undergoing knee arthroscopies. The 69 patients were randomised into three groups: group 1 received 150 ug clonidine intra-articularly, group two 150 ug clonidine intravenously and group three a placebo. Postoperative pain therapy was carried out with i.v. morphine hydrochloride. Pain scores and side-effects were documented for 24 h. RESULTS: There were no significant differences between the three groups in demographics, duration of operation, duration of anaesthesia, diagnoses or type of operation. The pain score at rest was significantly lower in group 1. In the first 20 min, the systemic concentration of clonidine was significantly higher in the intravenous group than in the intra-articular group. CONCLUSION: Intra-articular clonidine has a postoperative analgesic effect after knee arthroscopies due to a peripheral action.

Determination of 4-amino-m-cresol and 5-amino-o-cresol and metabolites in human keratinocytes (HaCaT) by high-performance liquid chromatography with DAD and MS detection.

A multi-step gradient HPLC system combined with DAD and MS detection has been developed for the determination of the oxidation hair dyes 4-amino-m-cresol (4-AC) and 5-amino-o-cresol (5-AC) and their metabolites in the alternative testing system human keratinocytes (HaCaT) cell culture. The culture medium induced by 3-methylcholanthrene (3-MC) was fortified with 4-AC or 5-AC and incubated for 24 h at 37 degrees C in order to produce metabolites. After several pre-cleaning steps, further cleaning was done by solid-phase extraction using C18 phenyl cartridges. Optimizing chromatographic conditions, a hybrid-based RP8 column was most suitable for the separation of the metabolites formed in HaCaT. Only one conjugation product, the N-acetylated derivative, could be identified for both 4-AC and 5-AC by LC/DAD/MS. The ionisation technique used for MS analysis was Atmospheric Pressure Ionization (API).

Determination of 4-amino-m-cresol and 5-amino-o-cresol by high performance liquid chromatography and fluorescence derivatization using fluorescamine.

For the determination of two oxidation hair dyes, 4-amino-m-cresol (4-AC) and 5-amino-o-cresol (5-AC), a sensitive isocratic high performance liquid chromatography (HPLC) method using the reversed phase mode was developed. The hair dyes were pre-column derivatized with fluorescamine prior to injection. Sensitivity could be improved 10-fold for 4-AC and 50-fold for 5-AC by fluorescence detection compared to UV detection. The limit of detection was 1 ng/injection for 4-AC and 100 pg/injection for 5-AC, respectively. For the determination of both compounds in aqueous biological matrices in order to simulate conditions for penetration studies with pig skin, a solid phase extraction procedure using C18 cartridges and acetonitrile (ACN) for elution could be developed. Average recovery was 83.4% with a coefficient of variation (CV) of 2.64% for intra-day assay and 3.20% for inter-day assay for 5-AC and 2.89% and 3.41% for 4-AC, respectively.

Determination of bone and tissue concentrations of teicoplanin mixed with hydroxyapatite cement to repair cortical defects.

A highly specific and sensitive isocratic reversed-phase high performance liquid chromatography (HPLC) method for the determination of the major component of teicoplanin in tissue is reported. Comparing fluorescamine and o-phthalaldehyde (OPA) as derivatizing agents, the derivative formed with the latter exhibits superior fluorescence intensity allowing detection of femtomole quantities. Pretreatment for tissue samples is by solid-phase extraction which uses Bakerbond PolarP C(18) cartridges and gives effective clean up from endogenous by-products. Linearity was given from 0.6 to 100 ng per injection. The coefficient of variation did not exceed 5.8% for both interday and intraday assays. It was found that when bone defects are repaired with a hydroxyapatite-teicoplanin mixture, the antibiotic does not degrade, even when it is in the cement for several months. The stability of teicoplanin in bone cement was determined fluorodensitometrically.

Localization of indoleamine 2,3-dioxygenase in human female reproductive organs and the placenta.

Indoleamine 2,3-dioxygenase (IDO) has been implicated in regulation of feto-maternal tolerance and protection against intracellular and extracellular pathogens. We have studied the expression of IDO in the human female reproductive tract and the placenta by immunohistochemistry. Endometrial glandular and surface epithelial cells showed increasing IDO expression during the course of the menstrual cycle. In term placenta, IDO was irregularly localized to the mesenchymal core and found in isolated areas of the syncytiotrophoblast. In first trimester pregnancy, IDO was not present in placental villi, but was present in glandular epithelium of the decidua, and there were distinctly positive cells scattered in the connective tissue, sometimes in conjunction with lymphoid aggregates. The endothelium of spiral arteries and of capillaries showed some, albeit no generalized, reactivity. IDO was also present in the epithelium of cervical glands and of Fallopian tubes. Specificity of antibody binding was confirmed by Western blot analysis. IDO mRNA was detected in first trimester decidua as determined by RT-PCR. IDO is secreted, as determined by analysis of cervical mucus by high pressure liquid chromatography for the presence of the tryptophan metabolite L-kynurenine, indicating IDO activity. Our results support the concept of IDO providing a mechanism of innate immunity protecting against ascending infections in the female reproductive tract.

The carcinostatic and proapoptotic potential of 4-hydroxynonenal in HeLa cells is associated with its conjugation to cellular proteins.

BACKGROUND: Previous studies have shown that the lipid peroxidation product 4-hydroxynonenal (HNE) acts as a cell growth modulator if used at low, physiological concentrations being strongly cytotoxic at higher concentrations for a number of cells. These effects of HNE also appeared to be mutually dependent on the effects of serum growth factors. The aim of this investigation was to study the concentration-dependent response of human cervical carcinoma (HeLa) cells in vitro with respect to the intracellular uptake of exogenous HNE, the cellular energy metabolism, DNA synthesis, overall gene expression and susceptibility to apoptosis. MATERIALS AND METHODS: MTT assay was applied as an index of energy metabolism and the replicative activity was quantitated by the 3H-thymidine incorporation assay. The occurence and intracellular distribution was studied with monoclonal antibodies directed against HNE-protein conjugates. Binding of HNE to serum proteins was determined with the same antibodies by Western blotting. Differential gene expression was studied by differential display RT-PCR while a novel photometric assay, denoted Titer-TACS, was used for in situ detection and quantitation of apoptosis in monolayer cell cultures. RESULTS: A physiological concentration of HNE (1 microM) had hardly any effect on the parameters of the replicative activity and the energy metabolism. No morphological changes were observed and the number of HNE-positive cells was not significantly different when compared to the untreated control cells, while most of the aldehyde appeared to be bound to serum proteins (albumin fraction). A ten-fold higher concentration (10 microM) was found to be cytostatic. Spindle-shaped cells with a picnotic nucleus were observed occasionally, as well as membrane blebs, which were HNE-positive. The number of HNE-positive cells was significantly increased compared both to the control cells and cells treated with 1 microM HNE, but in the presence of serum the effects of 10 microM HNE were negated due to its binding to the serum proteins. Finally, 100 microM HNE was cytotoxic for the HeLa cells. Most of the cells were picnotic, together with a few spindle-shaped or oval cells. The staining for HNE was diffuse and strong (90% of the cells were HNE-positive) while even binding of the aldehyde to serum proteins did not prevent its cytotoxic effects. This concentration of HNE caused acute stress response of the cells resulting in the decreased expression of several as yet unidentified genes. The altered pattern of gene expression was followed by programmed cell death, i.e. an increased number of apoptotic cells after treatment with low (1 and 10 microM) concentrations of HNE. A rebound effect was observed, i.e. a decrease of apoptotic cells after 24 hours followed by an overshooting increase after 48 hours. CONCLUSIONS: For HeLa carcinoma cells there appears to be a concentration range of HNE where it does not cause necrosis but preferentially apoptosis. At this concentration range HNE is cytochemically detectable within the cells as a protein conjugate. It is proposed that a possible differential sensitivity of cancer cells and their normal counterparts to the cytostatic activity of HNE should be explored.

Formation of 8-iso-PGF(2alpha) and thromboxane A(2) by stimulation with several activators of phospholipase A(2) in the isolated human umbilical vein.

We investigated the effects of the phospholipase A(2) (PLA(2)) activators calcium ionophore A 23187, hydrogen peroxide (H(2)O(2)), bradykinin (BK), histamine and noradrenaline (NA) on the 8-iso-prostaglandin (PG)F(2alpha) formation in the isolated human umbilical vein and the isolated rabbit ear. For comparison, the influence of these substances on the thromboxane A(2) (TXA(2)) release was also investigated. The release of total (esterified as well as free) 8-iso-PGF(2alpha), free 8-iso-PGF(2alpha) and TXB(2), the stable metabolite of TXA(2), was determined by specific enzyme immunoassays. The results show that bolus injections of 5.4 mmol H(2)O(2), 30 nmol A 23187, 10 nmol BK, 50 nmol histamine and 20 nmol NA caused an increased release of total 8-iso-PGF(2alpha) in the umbilical vein and the rabbit ear. A perfusion with H(2)O(2) at a final concentration of 0.3 mM also increased the release of this isoprostane. Increased formation of free 8-iso-PGF(2alpha) was induced by A 23187 injection and by both modes of H(2)O(2) administration, but not by the other treatments. Bolus injections of A 23187, BK and histamine induced an increased release of TXB(2) in both organs. Both modes of H(2)O(2) administration and NA showed no releasing effects. In conclusion, our results show that the substances used are able to stimulate the formation of 8-iso-PGF(2alpha) concurrently with the release of PGs. This effect might be of pathophysiological relevance in inflammatory and cardiovascular diseases in which an enhanced release of free radicals, BK, histamine or NA play an important role.

Characterization of prostanoid receptors mediating actions of the isoprostanes, 8-iso-PGE(2) and 8-iso-PGF(2alpha), in some isolated smooth muscle preparations.

We investigated the contracting actions of the isoprostanes (isoPs), 8-iso-prostaglandin (PG) F(2alpha) and 8-iso-PGE(2), in comparison to the effects of the thromboxane (TX) A(2)-mimetic U 46619 and the traditional prostaglandin PGE(2) in the isolated rat aorta, isolated rat gastric fundus and the isolated guinea-pig ileum. U 46619 and 8-iso-PGF(2alpha) caused contractions in the rat aorta and rat gastric fundus in a concentration-dependent manner, whereas these agonists showed no effects in the guinea-pig ileum. However, 8-iso-PGE(2) and PGE(2) caused contractions in all isolated organs used. The prostanoid TP-receptor antagonist SQ 29,548 (10 nM) significantly antagonized vasoconstrictions induced by the agonists used in the rat aorta. SQ 29,548 at a final concentration of 3 microM, but not at lower concentrations, significantly inhibited contractions induced by U 46619, 8-iso-PGF(2alpha) and 8-iso-PGE(2) in the rat fundus. Responses to PGE(2) were unchanged. The prostanoid EP(1)-receptor antagonist SC 51089 (3 microM) significantly inhibited contractions induced by 8-iso-PGE(2) and PGE(2) in the rat fundus and in the guinea-pig ileum. SC 51089 had no effect on responses to any of the agonists tested. Our results show that 8-iso-PGE(2), in contrast to 8-iso-PGF(2alpha), can also cause contractions by activation of the EP(1)-receptors in the rat gastric fundus and the guinea-pig ileum. The findings of the present study do not support the existence of a unique isoP-receptor in the tissues used.

Influence of polyunsaturated fatty acids on vasoconstrictions induced by 8-iso-PGF(2alpha) and 8-iso-PGE(2).

8-iso-PGF(2alpha) and 8-iso-PGE(2), which are released in vivo by free radical catalyzed peroxidation of arachidonic acid, are equipotent vasoconstrictors in vivo and in vitro. It is assumed that they exert this effect via activation of the thromboxane A(2) (TP) receptor or a TP-receptor-like isoprostane receptor. Increased levels of 8-iso-PGF(2alpha) have been detected in human cardiovascular diseases. It has been found that polyunsaturated fatty acids (PUFAs) have many beneficial effects in cardiovascular diseases, including antivasoconstrictor actions. Therefore, we investigated the influence of perfusions with eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and dihomo-gamma-linolenic acid (DGLA) at final concentrations of 3 and 30 micromol/l on vasoconstriction induced by 8-iso-PGF(2alpha), 8-iso-PGE(2) and the thromboxane A(2) mimetic U 46619 in the vasculature of the isolated perfused rabbit ear. Additionally, the effect of indomethacin (final concentration 3 micromol/l) on the effects of the PUFAs was investigated. Our results show that the PUFAs at a concentration of 30 micromol/l caused a significant inhibition of the vasoconstrictions induced by 8-iso-PGF(2alpha), 8-iso-PGE(2) and U 46619. Furthermore, it can be assumed that a part of the inhibitory effect of DGLA is due to the effect of a cyclooxygenase product, probably PGE(1), because indomethacin reduced the inhibitory effect of DGLA.

Reaction patterns of monoclonal antibodies to HLA-G in human tissues and on cell lines: a comparative study.

We compared the immunohistochemical reaction patterns of HLA-G-specific antibodies 87G, 4H84, G233, 16G1, and BFL.1 on human placentas under three different preparative conditions and on cryosections of other human tissues. Human and murine cell lines, either naturally expressing or transfected with HLA-G, were analyzed for their reaction patterns by immunocytochemistry and flow cytometry. Antibodies HCA2, TP25.99, W6/32 to classical HLA class I, anti-beta(2)-m and various non-HLA-G expressing cell lines were used as controls. The binding ability of the antibodies depends on the histotechnical procedure used. 4H84 and HCA2 bind to HLA-G despite aldehyde fixation and also paraffin embedding. 87G does not bind HLA-G in studies involving fixation with aldehydes. G233 labels HLA-G in aldehyde fixed but not paraffin embedded tissues. By immunocytochemistry HLA-G2 is merely detected with antibodies 4H84 and HCA2. MAb 16G1 binds to HLA-Gsol transfected cell lines only. The HLA-G specificity of mAb BFL.1 was considered as doubtful because it failed to react with most of the HLA-G transfected cell lines. Binding of 87G to the surface of monocytes or U-937 cells stimulated with IFN-gamma and GM-CSF is an Fc-receptor mediated phenomenon.

Influence of isoprostanes on vasoconstrictor effects of noradrenaline and angiotensin II.

The isoprostanes, 8-iso-prostaglandin F2alpha and 8-iso-prostaglandin E2, which are released in vivo by free radical-catalyzed peroxidation of arachidonic acid, are potent vasoconstrictors. Increased formation of 8-iso-prostaglandin F2alpha has been detected in human cardiovascular diseases, in which enhanced plasma levels of noradrenaline and angiotensin II have harmful vasoconstrictor effects. Therefore, we investigated the influence of perfusions with the thromboxane A2 mimetic, U 46619, and with the isoprostanes, 8-iso-prostaglandin F2alpha, 8-iso-prostaglandin E2, 8-iso-prostaglandin E1 and 8-iso-prostaglandin F3alpha, on the vasoconstrictor effects of noradrenaline and angiotensin II in the isolated perfused rabbit ear. Our results demonstrate that perfusions with U 46619, 8-iso-prostaglandin E2 and 8-iso-prostaglandin F2alpha, at a subthreshold concentration (30 nM), amplified the vasoconstrictions induced by noradrenaline or angiotensin II significantly. In addition, the results show that U 46619, 8-iso-prostaglandin F2alpha, 8-iso-prostaglandin E2 and 8-iso-prostaglandin E1, which were applied as a bolus, induced much more pronounced vasoconstrictions than prostaglandin F2alpha, prostaglandin E2 and prostaglandin F3alpha. Prostaglandin E1 and 8-iso-prostaglandin F3alpha, showed no effects. In conclusion, it can be assumed that the powerful vasoconstrictions induced by 8-iso-prostaglandin E2 and 8-iso-prostaglandin F2alpha and their potentiating effects on vasoconstrictions induced by noradrenaline or angiotensin II might be of pathophysiological relevance in cardiovascular diseases.

Vascular effects of isoprostanes after endothelial damage.

The isoprostanes, 8-iso-PGF2alpha and 8-iso-PGE2, are powerful vasoconstrictors in vitro and in vivo. Increased formation of 8-iso-PGF2alpha was detected in atherosclerotic patients. In this disease endothelial injuries appear, followed by a reduced release of nitric oxide (NO). Our results show that the vasoconstrictor effects of 8-iso-PGF2alpha as well as of 8-iso-PGE2 were amplified after endothelial damage of the vasculature of the isolated perfused rabbit ear. Also vasoconstrictions induced by both isoprostanes were amplified by perfusions with the NO-synthase blocker N(G)-nitro-L-arginine methylester (L-NAME) at a final concentration of 100 micromol/l. Therefore, we can assume that the amplification of the vasoconstrictor effects of the isoprostanes used after damage of endothelium might be mainly due to the reduced formation of NO.

Biocompatibility of heparin and low-molecular mass heparin in a new implantable medication pump.

The present paper deals with the evaluation of normal heparin and low-molecular mass heparin as to their use in a new implantable medication pump. Latest versions of different Pharmacopoeias recommend techniques, which require large sample volumes for aqueous injectable drug solutions. Therefore, a commercially available enzymatic kit was used to determine heparin activity. As this test is designed for heparin plasma levels, transformation of the chromophore occurred only after adding heparin AT III complex, which is usually present in plasma. The applied test was found to be suitable regarding reproducibility, selectivity and sensitivity. There was no loss of enzyme activity of both normal heparin and low-molecular mass heparin within an investigation period of 8 weeks; thus, no decomposition products are to be expected and the biocompatibility of pump materials with both tested heparin preparations is evident.

Solid-phase synthesis of peptide nucleic acid (PNA) monomers and their oligomerization using disulphide anchoring linkers.

A new simple solid-phase method has been developed for synthesizing Boc-protected peptide nucleic acid (PNA) monomers. An immobilized backbone 3 was built on Expansin resin using an ester disulphide handle: 2-hydroxypropyl-dithio-2'-isobutyric acid (HPDI). The base acetic acids of thymine 5, Z-cytosine 9, Z-adenine 12, and 6-O-benzyl guanine 17 were prepared and coupled to the immobilized backbone. The HPDI handle was cleaved under mild conditions by cyanolysis or assisted hydrolysis with tris(2-carboxyethyl)phosphine (TCEP) to give undamaged PNA monomers. These monomers were coupled to form oligomers by solid-phase method with another disulphide linkage: aminoethyldithio-2-isobutyric acid (AEDI) grafted on an amino-functionalized TentaGel resin, using in situ neutralization and TBTU as activating reagent. Final cleavage of the AEDI linker gave PNA bearing a cysteamide residue that could be useful for optimizing PNA properties. Oligomers of up to 16 residues long were assembled.

Determination of SDZ ICM 567 in blood and muscle microdialysis samples by microbore liquid chromatography with ultraviolet and fluorescence detection.

Fast, simple and accurate methods for the determination of SDZ ICM 567, the 7-methoxy derivative of tropisetron, in microdialysates have been developed. Sampling by microdialysis from freely moving rats in the portal and jugular vein offers a new technology for pharmacokinetic studies by direct and continuous measurement of unbound drug concentrations with time. SDZ ICM 567 can be identified in small sample volumes of dialysates on a microbore high-performance liquid chromatography column-switching system with ultraviolet detection. In addition, determination of SDZ ICM 567 by fluorimetric detection has been developed for muscle microdialysates from rats. [14C]SDZ ICM 567 was used as reference substance for the estimation of the amount of substance transferred through the dialysis membrane. The radioactive measurement (RA) gave the recovery information, whereas the liquid chromatographic method detected the sum of [14C]SDZ ICM 567 and dialyzed SDZ ICM 567.

Phenotypic comparison of natural killer cells from peripheral blood and from early pregnancy decidua.

In this descriptive flow cytometric study we analyzed the phenotype of human large granular lymphocytes from the decidua (DLGL) of first-trimester pregnancy. Expression of CD56 at high density on DLGL suggests a relationship to the small CD56bright+ subpopulation of peripheral blood natural killer (PBNK) cells. In comparison, these cell types differ in respect to the expression of a variety of adhesion molecules and receptors implicated in homing, migration and activation. In contrast to CD56bright+ PBNK cells, DLGL were still brighter for CD56 and show higher expression for CD29 and CD45RO. Less expression was found for CD15s, CD43, CD44, CD45RA, CD62L and HLA-DR. CD11a to c and CD18 were distributed in bimodal form on DLGL, part of the cells being negative. In summary, we found considerable differences between the cell surface marker profiles of DLGL and PBNK cells (subpopulations of the latter being separately analyzed).