A new method to continuously change the buffer conditions in samples prior to electrophoresis by counter-current dialysis has been developed using a hollow fiber membrane module (VariPerm M, bitop, Witten). It is demonstrated that the collected fractions of a column eluate after hydrophobic interaction chromatography cannot be analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) due to the high content of ammonium sulfate. In contrast, when the hollow fiber membrane module is used for on-line desalting of the eluate during chromatography, it is possible to detect distinct bands on SDS-gels even after sample concentration by evaporation. Moreover, adding polyethyleneglycol 20,000 to the counter-current dialysis buffer the hollow fiber membrane module is suitable for combined on-line sample desalting and sample concentration of living organisms during free flow electrophoresis (FFE): a culture of Nitrosomonas eutropha which could not significantly be detected by FFE without conditioning was successfully analyzed when concentrated and desalted on-line during sample application.

Ammonium Sulfate↗

PubMedDate not supplied

Alpha-receptor constriction induced by atrial fibrillation during maximal coronary dilatation.

The mechanism of coronary vasoconstriction induced by atrial fibrillation during maximal coronary dilatation was studied in 19 chloralose-urethane anesthetized dogs. Maximal coronary dilatation was achieved by carbochromene (5 mg/kg i.v.) or dipyridamole (0.2 mg/kg i.v.). Left circumflex coronary blood flow was measured with an electromagnetic flowmeter. Atrial fibrillation was compared with rhythmic atrial pacing at similar heart rates (207 +/- 12 vs. 204 +/- 12 beats/min). During maximal coronary dilatation, coronary resistance was 0.38 +/- 0.05 mm Hg X min X 100 g/ml (RU) at sinus rhythm, 0.41 +/- 0.06 RU at atrial pacing, and 0.52 +/- 0.07 RU at atrial fibrillation, that was significantly (p less than 0.005) higher than during sinus rhythm and atrial pacing. Accordingly, coronary oxygen extraction was 14 +/- 1% at sinus rhythm, 17 +/- 1% at atrial pacing (p less than 0.005 vs. sinus rhythm) and 27 +/- 2% at atrial fibrillation (p less than 0.001 vs sinus rhythm and atrial pacing). Beta-adrenoceptor blockade with propranolol (1 mg/kg i.v.) did not prevent this coronary vasoconstrictive effect. Following alpha-blockade with phenoxybenzamine (10 mg/kg i.v.), however, coronary resistance was 0.52 +/- 0.08 RU during sinus rhythm, 0.54 +/- 0.10 RU during atrial pacing and 0.57 +/- 0.09 RU during atrial fibrillation. The data suggest coronary vasoconstriction induced by atrial fibrillation mediated by an alpha-adrenoceptor mechanism.

Animals↗

PubMedDate not supplied

Expression of cytokine receptors in the placenta in term and preterm labour.

Elevated uterine concentrations of interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumour-necrosis factor-alpha (TNF-alpha) are suspected to cause increased prostaglandin release from gestational tissues, but little information is available about the expression pattern of cytokine receptors in these tissues. In this study, cytokine receptor positive cells in frozen tissue sections of placentae (n=70) and fetal membranes (n=50) were identified by immunohistological staining with monoclonal antibodies specific for IL-6 receptor, TNF receptors I and II, and IL-1 receptor I. Both subunits of the IL-6 receptor (gp130 and gp80) as well as TNF receptors I and II were expressed by fetal endothelial cells within placental villi, while IL-1-receptor I was detected exclusively in stromal cells of the maternal decidua. The IL-1 receptor I and TNF receptors I and II were expressed in both uterine quiescence and labour, irrespective of gestational age. Immunoreactivity of the gp130 subunit of the IL-6 receptor was found also throughout pregnancy, while the appearance of the gp80 subunit correlated with the presence of term and preterm labour. In case of preterm labour, expression of the gp80 subunit was predominantly detected in the absence of intrauterine infection. Therefore, it is concluded that the de novo expression of the gp80 subunit and consequently the appearance of entire IL-6 receptors in the placenta is associated with spontaneously occurring labour at term and also with preterm occurring labour in the absence of inrauterine infection.

Adult↗

PubMedDate not supplied

Ovarian cancer antigen CA 125 enhances the invasiveness of the endometriotic cell line EEC 145.

OBJECTIVE: Ovarian cancer antigen CA 125 is used widely for diagnostic and therapeutic evaluation of ovarian cancer. Although the initial description of CA 125 was in 1981, its function is still unknown. In collagen gel system the invasiveness of the endometriotic cell line EEC 145 is enhanced by a proteinaceous factor in peritoneal fluid mimicking molecular features of CA 125. Therefore we hypothesize that this factor is CA 125. METHODS: The influence of heat-treated peritoneal fluid on the invasiveness of the endometriotic cell line EEC 145 and several human carcinoma cell lines (EJ28, RT112, AN3 CA, RL95-2, HEC-1-A, HeLa, MCF7, T-47D, and SK-OV-3) was investigated in a collagen gel invasion assay by addition of (1); 10% heat-treated peritoneal fluid, (2); peritoneal fluid after immunoprecipitation with antibodies directed to CA 125, and (3); purified CA 125. RESULTS: The invasion index of the endometriotic cell line EEC 145 significantly increased from 4.78 +/- 0.98 to 6.57 +/- 1.58 (P = .0001) by addition of 10% heat-treated peritoneal fluid to the culture medium. The invasion-promoting effect of peritoneal fluid was abolished by reduction of the CA 125 concentration through immunoprecipitation and was mimicked in a dose-dependent manner by addition of CA 125 to the culture medium. The invasiveness of the investigated human carcinoma cell lines was not affected by heat-treated peritoneal fluid of CA 125. CONCLUSION: The results are consistent with the hypothesis that CA 125 can influence invasiveness in a benign endometriotic cell line.

Ascitic Fluid↗

PubMedDate not supplied

[Intercellular adhesion molecular 1, a diagnostic serum marker in chorioamnionitis, pre-eclampsia and HELLP syndrome].

INTRODUCTION: Chorioamnionitis and pregnancy-induced hypertension both are extremely feared complications of human pregnancy. Activation or disturbance of normal endothelial cell function may be involved in the pathogenesis of both kinds of disease. The aim of our study was to compare the diagnostic value of soluble intercellular-adhesion-molecule-1 (ICAM-1) with that of C-reactive protein (CRP) and white blood cell count for the detection of chorioamnionitis in patients with preterm labor. In addition, we examined if concentrations of ICAM-1 were also increased in case of preeclampsia or HELLP-syndrome. MATERIALS AND METHODS: ICAM-1, CRP and leucocyte count were estimated in 50 cases of normal term delivery, 97 cases of uncontrollable preterm labor, 16 cases of preeclampsia and 9 cases of HELLP-syndrome before delivery. RESULTS: From 97 women delivering preterm, chorioamnionitis was histologically confirmed for 48 women. Maternal serum levels of ICAM-1 (p < 0.001), CRP (p < 0.001) and leucocyte count (p < 0.02) were significantly higher in the group of preterm delivering patients (< 37 weeks gestation) with histologically confirmed chorioamnionitis in comparison to preterm delivering patients in the absence of chorioamnionitis. In the group of patients delivering preterm (< 37 weeks gestation) because of preeclampsia or HELLP-syndrome, ICAM-1 (p < 0.001), as well as CRP (p < 0.006) concentrations were also significantly increased in comparison to patients delivering preterm in the absence of chorioamnionitis. CONCLUSIONS: Elevated levels of ICAM-1 in the serum of pregnant women may be considered as an important risk factor for the development of complications in pregnancy associated with inflammatory induced changes of maternal endothelial cell functions.

Adult↗

PubMedDate not supplied

[Disturbed utero-tubal transport in hysterosalpingoscintigraphy as a predictive functional test for IVF therapy].

Hysterosalpingoscintigraphy (HSSG) is a simple method to evaluate the transport function of uterus and fallopian tubes. There is a quick uptake of radionuclides into the uterus and a transport to the side bearing the dominant follicle in 70 % of the patients in the late follicular phase of the cycle. Uptake and transport of the immotile radionuclides imitate the directed sperm transport through the female genital tract at the time of ovulation. 214 of 796 infertility patients with proven patency of fallopian tubes (27 %) showed only an uptake of the particles without a transport towards the fallopian tubes (negative HSSG). In these patients no spontaneous pregnancy occurred and pregnancy rate remained low by the means of timed intercourse or insemination. Indeed, the pregnancy rate (8.4 %) was significantly lower compared with the patients who became pregnant by timed intercourse, insemination or spontaneously and had a positive HSSG before (15 %; p=0.001). However, the pregnancy rate that could only be achieved by methods of ART was significantly higher in the group of patients with negative HSSG (57 % vs. 25 %, p=0.05). Our data suggest that HSSG is a new method to evaluate the integrity of the inner genital tract's transport function, especially in patients suffering from idiopathic infertility. Impaired transport function (negative HSSG) should be considered as an indication for IVF-treatment.

Fallopian Tubes↗

PubMedDate not supplied

Differences in acidogenicity of S. sobrinus and S. rattus are linked to the catalytic efficiency of the glycolytic key enzyme phosphofructokinase.

This contribution describes the biochemical properties of two catalytically different phosphofructokinases (PFKs) purified from Streptococcus rattus LB 2 (PFK-rat) and Streptococcus sobrinus OMZ 65 (PFK-sob), respectively. Steady-state kinetics revealed K(M) = 0. 8 mM for PFK-rat and K(M) = 0.08 mM for PFK-sob for F-6-P as the substrate. The enzymes also differ in their pH profiles: whereas the highest activity of PFK-rat was measured at pH = 8.0, the optimum pH of PFK-sob was at pH = 7.0. In addition, compared to PFK-sob, PFK-rat was more sensitive against the allosteric inhibitor ATP. PFK catalyzes a committed step of glycolysis, the main acid producing catabolic pathway. Thus, the catalytically more efficient enzyme isolated from S. sobrinus OMZ 65, especially at low pH, could explain the comparably high acidogenicity of this strain.

Bacterial Proteins↗

PubMedDate not supplied

The tumor markers TPA, TPS, TPACYK and CYFRA 21-1 react differently with the keratins 8, 18 and 19.

The commercially available tumor marker tests TPA, TPS, TPACYK and CYFRA 21-1 react with simple epithelium keratins. From clinical studies it can be deduced that the pattern of keratin recognition must be different for each of these tests. We therefore studied the reactivity of the keratin fragment combinations K8/K18 and K8/K19 in the different tests and determined the reactivity of the corresponding soluble antibodies with purified keratin 8, 18 and 19 in immunoblots. TPS and CYFRA 21-1 were found to distinguish clearly between the keratin fragment combinations K8/K18 (TPS) and K8/K19 (CYFRA 21-1). TPA and TPACYK reacted with both combinations, however, with different intensities. On immunoblots the CYFRA 21-1 antibodies reacted exclusively with K19, whereas the antibodies of the other assays reacted with at least 2 of the keratins investigated.

Antibodies↗

PubMedDate not supplied

Molecular systematics of the nepetoideae (family Labiatae): phylogenetic implications from rbcL gene sequences.

Total DNA was extracted from 41 species (20 genera) of the subfamily Nepetoideae (family Labiatae). Using rbcL-specific primers, the rbcL gene wsa amplified by polymerase chain reaction (PCR) and sequenced directly. RbcL sequences were evaluated with character state (maximum parsimony; PAUP) and distance methods (neighbour-joining; MEGA). In agreement with classical systematics all taxa studied cluster within the Nepetoideae and are clearly distinguished from members of the subfamily Lamioideae. A number of distinctive clades are apparent within the Nepetoideae: I-Collinsonia, II-Lavandula, III-Agastache, Glechoma, IV-Satureja, Hyssopus, Dracocephalum, V-Nepeta, VI-Hormium, VII-Prunella, VIII-Melissa, Ocimum, IX-Monarda, Mentha, X-Origanum, Thymus, XI-Salvia, XII-Rosmarinus, and XIII-Perovskia. At least five main branches representing the clades I, II, III to VII, VIII, and IX to XIII respectively, can be distinguished within the Nepetoideae studied. They might be considered representing the tribes (according to Cantino, 1992) Elsholtzieae (I). Lavanduleae (II), and Mentheae (III-XIII). The tribe Mentheae needs to be subdivided into at least three main groups (clades III-VII, VIII and IX-XIII). Majorana hortensis which is often classified as Origanum hortensis does not cluster with Origanum and deserves a genetic status of its own.

Base Composition↗

PubMedDate not supplied

MIA, a novel serum marker for progression of malignant melanoma.

MIA was isolated previously as a small soluble protein secreted from malignant melanoma cell lines in vitro. Highly restricted expression patterns in melanocytic tumors were identified in vivo. We therefore quantitated serum levels of MIA protein by means of a non-radioactive ELISA and investigated whether MIA provides clinically relevant parameters in patients with malignant melanomas. Here we report enhanced MIA serum levels in 13% and 23% of patients with stage I and II disease, respectively, and in 100% with stage III or IV disease. Response to therapy in stage IV disease correlated with changes in MIA serum levels and surgical removal of metastases led to normalization of serum values. Repeated measuring of sera from 350 patients with a history of stage I or II melanoma during follow-up, we detected 32 patients developing positive MIA values. At the time of serum analysis 15 of them had developed metastases and one presented with metastatic disease 6 months later. In conclusion, MIA represents a novel serum marker for systemic malignant melanoma revealing the highest sensitivity and specificity among currently available markers.

Adult↗

PubMedDate not supplied

Surgical closure of atrial septal defects. Results of the circumclusion technique.

Explore the source record for details and available documents.

Adolescent↗