Low maternal serum concentrations of mannose-binding lectin are associated with the risk of shorter duration of pregnancy and lower birthweight.

Chronic inflammation has been implicated as the underlying mechanism responsible for the pathophysiology of preterm labour. Mannose-binding lectin (MBL) plays a central role in the innate immune response and is thus an important component of the first line of defense. The aim of this study was to investigate whether serum concentrations of MBL correlated with the incidence of preterm birth and low birthweight in a cohort of women with signs of threatened preterm birth. A cohort of 60 patients who presented with regular contractions and/or short cervix (group A) between 24 and 32 weeks of gestation and 20 healthy controls (group B) who had no pregnancy complications and delivered at term were recruited into a prospective study. The following outcomes were recorded: presence of preterm labour and birthweight in all patients. MBL and high sensitivity C-reactive protein levels were measured in all serum samples. The serum concentrations of MBL were significantly reduced in patients with threatened preterm labour (Group A), compared to the control Group B. Furthermore, infants born to Group A mothers with MBL deficiency (n = 13, MBL ≤100 ng/mL) had significantly lower birthweights, compared to those born to Group A women with normal MBL serum concentrations (P < .0001). Our small cohort study demonstrated a strong association between MBL deficiency and preterm delivery, and associated low birthweight. MBL deficiency could thus be considered an important risk factor for preterm birth.

[The role of T-regulatory lymphocytes in pathogenesis of preterm delivery]. / Úloha T-regulacních lymfocytu v patogenezi predcasného porodu.

OBJECTIVE: To describe the role of T-regulatory lymphocytes in pathogenesis of preterm delivery. SETTING: Department of Obstetrics and Gynecology, General University Hospital and 1st Medical Faculty, Charles University, Prague. METHOD: T-regulatory lymphocytes modulate the immune system, secure the tolerance to own antigens and prevent autoimmune disease. During pregnancy is maternal immunity in contact with the semi-allogeneic fetus due to the fetomaternal crosstalk. It seems that maternal immunity and T-regulatory lymphocytes have an effect on premature birth and other pregnancy pathologies. According to the latest data, their role in the immunomodulation of pregnant women seems to be very significant, although we still do not understand many mechanisms.

Serum copper as a marker of inflammation in prediction of short term outcome in high risk patients with chronic heart failure.

BACKGROUND: Oxidative process and inflammation are regarded as important factors in the pathogenesis of chronic heart failure. Our study was aimed at investigating the prognostic value of serum copper levels in high risk subjects with chronic heart failure. METHODS: Serum copper levels and other prognostic indicators were determined in the group of 60 patients with chronic heart failure due to ischemic heart disease: 30 consecutive subjects with acute decompensation of chronic heart failure (acute group A) and 30 patients with chronic stable heart failure (group B). Patients were followed prospectively 12 months. Primary end-point was the mean time to death and/or heart failure hospital admission. RESULTS: The mean time to death was in the group A 279.4+/-18.9 days and 351.7+/-13.6 days in the group B (p<0.0001). Cox proportional hazard model revealed that the time to death for all subjects (n=60) was affected by cardiothoracic ratio (p<0.001). The time to combined end-point death or hospital admission was affected by serum copper concentration (p<0.0001). CONCLUSION: Serum copper levels predicted short term outcome in high risk patients with chronic heart failure.

Simultaneous determination of molybdenum(VI) and tungsten(VI) and its application in elemental analysis of polyoxometalates.

Spectrophotometric determination of molybdenum(VI) and tungsten(VI) with application of Artificial Neural Networks is proposed and it was applied for elemental analysis of solid polyoxometalates. Better results in comparison with previously those achieved by previous published method were demonstrated. MALDI-TOF Mass Spectrometry was tested for possible determination of molecular weight of polyoxometalates utilizing different matrices. Phenomena observed during desorption-ionisation processes are discussed. LDI-TOF MS was found to be suitable for the determination of Mo:W ratio in polyoxometalates as a rapid screening method to follow synthetic procedure.

Hyperlipoproteinemia impairs endothelium-dependent vasodilation.

Atherogenic lipoproteins can cause endothelial dysfunction in the initial stage of atherogenesis. In our study we examined 134 patients with defined hyperlipoproteinemia (non-HDL cholesterol>4.1 mmol/l or triglycerides>2.5 mmol/l or taking any of lipid lowering drugs)--94 men and 40 women. The subgroup of controls of comparable age contained 54 normolipidemic individuals--30 men and 24 women. Patients with hyperlipoproteinemia revealed significantly lower ability of endothelium-dependent flow-mediated vasodilation (EDV) measured on brachial artery (4.13+/-3.07 vs. 5.41+/-3.82 %; p=0.032) and higher carotid intima media thickness than normolipidemic controls (0.68+/-0.22 vs. 0.58+/-0.15 mm; p=0.005). In regression analysis, EDV correlated significantly with plasma concentrations of oxLDL (p<0.05) HDL-cholesterol (p<0.05), Apo A1 (p<0.05), ATI (p<0.01) and non-HDL cholesterol (p<0.05). Patients with hyperlipoproteinemia showed higher plasma levels of oxLDL (65.77+/-9.54 vs. 56.49+/-7.80 U/l; p=0.015), malondialdehyde (0.89+/-0.09 vs. 0.73+/-0.08 micromol/l; p=0.010) and nitrites/nitrates (20.42+/-4.88 vs. 16.37+/-4.44 micromol/l; p=0.018) indicating possible higher long-term oxidative stress in these patients.

[Ferritin, oxidative stress and coronary atherosclerosis]. / Feritin, oxidacní stres a koronární ateroskleróza.

BACKGROUND AND AIM: In the recent years several studies showed the association between body iron stores, represented by serum ferritin, and atherosclerosis. It was proposed that iron bound to ferritin catalyzes the formation of highly reactive forms of oxygen free radicals which subsequently cause the oxidative modification of atherogenic lipoproteins. Aim of our study was to compare serum ferritin concentrations and certain markers of oxidative stress in patients with and without coronarographically assessed coronary vascular disease. METHODS AND RESULTS: Measurements were performed in 216 subjects at the age of 35-60 years. The patient group included 76 patients with coronarographically assessed coronary vascular disease (CVD) (mean age 51.16 +/- 5.713 years) and 140 healthy controls (mean age 50.21 +/- 5.331 years). The plasma concentration of ferritin was higher in patients (169.04 +/- 63.899 micrograms/l) than controls (87.70 +/- 41.394 micrograms/l), p < 0.001. The group of patients revealed significantly lower plasma concentrations of anti-oxLDL antibodies, nitrites/nitrates, tocopherol and high density lipoprotein cholesterol (HDL-cholesterol) than controls; on the contrary patients had significantly higher concentrations of hemoglobin, thrombocytes and triacylglycerols. In the whole cohort of investigated subjects, ferritin correlated positively with retinol, body mass index (BMI), total-cholesterol, triacylglycerols, low density lipoprotein cholesterol (LDL-cholesterol), blood glucose, creatinine, uric acid, alaninaminotransferase (ALT), aspartateaminotransferase (AST), hematocrite, erythrocytes, with occurrence of CVD and with sex. Inverse correlation was observed between ferritin and HDL-cholesterol. CONCLUSIONS: Our observations are consistent with the hypothesis that high stored iron levels, measured by serum ferritin concentrations, may contribute to the oxidative stress and thus elevate the risk for development of CVD.

Experience with the iodine-123 and technetium-99m labelled anti-granulocyte antibody MAb47: a comparison of labelling methods.

Four different methods of radiolabelling the anti-granulocyte monoclonal antibody MAb47 were compared and their influence on diagnostic value studied. The best clinical images were obtained following labelling with iodine-123 by the Iodogen method and direct labelling with technetium-99m after tris-(carboxyethyl)-phosphine treatment of MAb47 to achieve disulphide bridge reduction. 99mTc labelling using a specific ligand (MAb47-mtp), or a second method involving direct reduction with mercaptoethanol, led to an increased background activity in clinical studies, thus impeding the diagnosis of chronic disease. Fresh infections were clearly localized by all four preparations. The elimination of the activity from the blood was slower in the case of the iodinated MAb47, while the collected urine samples showed an excretion of about 10% of the injected activity per day independent of the labelling method. The results in terms of sensitivity and specificity were rather similar for all labelling methods and ranged from 90% to 99%.

An experimental immunoscintigraphic study with anti-CEA monoclonal antibody (DG2).

Monoclonal antibody D11-DG2 (DG2) against carcinoembryonic antigen (CEA) was examined for suitability for radioimmunodetection of human tumors grown in nude mice. Antibodies DG2 and a control antibody of the same IgG1 subclass were labeled with 131I and injected into mice bearing one of three types of CEA-containing tumors (cell lines LS 174T, HT-29 and Rec S) and/or a CEA-negative tumor (Rec R). Gamma-camera imaging and distribution studies revealed that CEA-containing tumors selectively accumulate DG2 but Rec R does not. As the tumors differ in CEA-content, the highest accumulation of 131I-DG2 (corresponding to the best scintigraphic imaging) was found in LS 174T tumors, intermediate in Rec S and lowest in HT-29 tumors. The mean tumor-to-blood ratios on the sixth day after antibody administration were 4.6, 3.2, and 2.1, respectively, in the control experiments the value of this parameter was always lower than 1. The results showed the applicability of DG2 for immunoscintigraphic studies in patients. Furthermore, a positive correlation was found between the uptake of anti-CEA antibody and CEA-content in the tumors.

Distribution, kinetics and immunoscintigraphy of 131-I labelled intact antifibrinogen-fibrin antibody (AbFbg) and its F(ab')2 fragment.

131-I-labelled anti fibrin-fibrinogen antibody (AbFbg) was compared with its F(ab')2 fragment in distribution studies and by immunoscintigraphy with a view to tumour visualization in tumour bearing rats. The distribution studies indicated that the intact antibody is more concentrated in tumour tissue than the F(ab')2 fragment. By 168h after injection, when tumour-to-tissue ratios were highest in the majority of tissues, the tumour concentration of intact antibody was 3 to 4 times that of the F(ab')2 fragment. The intact antibody is more suitable than the F(ab')2 fragment for tumour imaging especially in the abdominal region where the highest tumour-to tissue ratios were obtained with intact antibody in liver, spleen, intestines and kidneys.

[Immunoscintigraphy in experiments and clinical practice: its possibilities, advantages and limitations]. / Imunoscintigrafie v experimentu, klinice, její moznosti, výhody a omezení.

The authors present their experience gained in preparing, isolating and labeling antibodies with radionuclides for the purpose of using them in immunoscintigraphy. The experimental part includes results obtained with different labeled antibodies and their F/ab/2 fragments in distribution studies, involving also immunoscintigraphic imaging of tumors. The clinical part presents results of immunoscintigraphy obtained with the commercial antibody kits Iodomab and Imacis in patients with tumors of the digestive tract.

Antiferritin antibodies in immunoscintigraphic detection of human tumor xenografts.

Affinity-purified antibodies against human placental ferritin and their F(ab)2 fragments labeled with 131I were examined for suitability for radioimmunodetection of ferritin-containing tumors. The nude mouse model (BALB/c, nu/nu) with xenografts of HeLa cell tumors and human adenocarcinoma of the rectum (with proven ferritin content) was used. Gamma-camera imaging and tissue distribution studies revealed that both kinds of tumor selectively accumulate antiferritin antibodies and their fragments. In large necrotic tumors nonspecific uptake of radiolabeled normal IgG occurred, but otherwise there was no tumor localisation. This study, in accordance with the literature, confirms the utility of antiferritin antibodies for the detection of human tumors in an animal model.

Identification of leukemia cell-derived inhibitory activity (LIA) in conditioned media from human myeloid leukemic cell line ML-2.

Conditioned media from the human myeloid leukemic cell line ML-2 contain a factor that inhibits the entry of normal CFU-GM into S phase of mitotic cycle as measured by the 3H-TdR suicide technique. This factor was detected in conditioned media prepared by incubating 5 X 10(6) ML-2 cells/ml or 1 X 10(6) ML-2 cells/ml in serum-free RPMI for 5 or 24 hours respectively, and was isolated by ultrafiltration through an XM 300 Diaflo membrane followed by chromatography on Sepharose 6 B. Ferritin, prepared from human placenta, had the same inhibitory effect on CFU-GM. Antibodies against human placental ferritin completely inactivated the inhibitory effect of both human placental ferritin and the factor released from ML-2 cells. The inhibitory activity produced by the cell-line ML-2 was considered as LIA (leukemia cell-derived inhibitory activity) earlier found in HL-60 cell line and AML and CML cells.

Ferritin-bearing lymphocytes in Hodgkin's disease.

In view of the reported association of Hodgkin's disease (HD) and ferritin, ferritin-bearing lymphocytes were followed during 2-year period in 79 HD patients. Indirect immunofluorescent method was used to evaluate the percentage of ferritin positive cells. In 22 untreated patients a high percentage of ferritin-bearing circulating lymphocytes (mean value 37.3%) was found. In regard to the extent of the disease higher values were found in clinical stage III and IV (mean value 40.6%) as compared to the stage I and II (mean value 26.2%). Similarly, 17 patients in relapse and with disease progression had mean values 41%. These proportions of cells were significantly lower in 44 patients in complete remission with mean value of 8.7% (60 examinations). In 30 healthy controls the mean value was 1.4%. Repeatedly performed examinations of ferritin-bearing lymphocytes during the follow-up period in 17 patients showed to be an important prognostic tool. A negative correlation of ferritin-bearing lymphocytes with E-rosette-forming cells was found. Iron content in peripheral blood lymphocytes was confirmed cytochemically after pre-incubation with antiferritin antibody. The results support the presumed role of ferritin in impaired cellular immunity in HD and suggest diagnostic and prognostic value of the examination of ferritin-bearing lymphocytes in HD.

Monoclonal antibodies against human urinary bladder carcinomas: selectivity and utilization for gamma scintigraphy.

Mouse monoclonal antibodies to human urinary bladder carcinoma cells have been examined by indirect membrane immunofluorescence using a panel of 27 human cell lines. Two of the monoclonal antibodies, 7E9 (IgG3) and S2C6 (IgGl), were found to distinguish between urinary bladder carcinoma cells and normal urothelium. The third monoclonal antibody, T24.06.5(IgGl), discriminated among cell lines of urothelial and non-urothelial origin but did not distinguish between urinary bladder carcinoma and normal urothelial cells. None of the of the antibodies was found to be strictly selective, and occasional cross-reactions with unrelated cell types were observed. The monoclonal antibody 7E9, showing the highest degree of selectivity, was further examined by an indirect immunoperoxidase technique on frozen tissue sections from 19 patients. The antibody reacted with all (7/7) bladder carcinomas examined and gave negative results with control normal bladder mucosa (0/8) and unrelated tumor tissue (0/4) sections. The 7E9 antibody was purified by protein A affinity chromatography, labeled with 131I and used for gamma-scintigraphy in nude mice xenografted with human urinary bladder carcinoma T24. The 7E9 antibody was capable of locating the T24 xenografts in nude mice; it localized preferentially in the T24 tissue compared to normal mouse tissues. The T24 xenografts could not be detected by gamma-scintigraphy with 131I-labeled monoclonal antibody against human mammary carcinoma cells and two other control antibodies. Likewise the 131I-labeled 7E9 antibody was not capable of locating human mammary carcinoma xenografts in nude mice.

Tumour localization of radiolabelled monoclonal antibody in mice bearing human urinary bladder (T24) carcinoma xenografts.

Selectivity of mouse monoclonal antibody 7E9 (IGG3) directed against human urinary bladder carcinoma cells has been examined by indirect membrane immunofluorescence, using a panel of 31 human cell lines. The 7E9 monoclonal antibody discriminated between urinary bladder carcinoma cells and normal urothelium or cells of non-urothelial origin, although occasional reactions with bladder carcinoma-unrelated cell types were observed. The 7E9 antibody was purified by protein A affinity chromatography, labeled with 131I and used for gamma scintigraphy in nude mice xenografted with human urinary bladder carcinoma T24. The 7E9 antibody was capable of locating the T24 xenografts in nude mice; it localized preferentially in the T24 tissue compared to normal mouse tissues. The T24 xenografts could not be detected by gamma scintigraphy with 131I-labelled monoclonal antibody against human mammary carcinoma cells and two other control antibodies. Likewise, the 131I-labelled 7E9 antibody was not capable of locating human mammary carcinoma xenografts in nude mice.

Immunoscintigraphy and distribution study of experimental tumours using 131I-labelled anti-fibrinogen antibodies.

131I-labelled antibodies to the fragment E of human fibrin, anti-rat fibrinogen, non-immune rabbit IgG and 67Ga-citrate were used for scintigraphy and distribution studies in experimental tumours in rats. The best visualisation was achieved with 131I-labelled anti-rat fibrinogen antibodies at intervals longer than 48 h after administration. Tumour to tissue ratios found in distribution studies performed formed more than 48 h after administration were higher in most examined tissues when using 131I-labelled antibodies than those obtained with 67Ga-citrate.

Lymphocyte nucleoli activation and changes of the mononuclear phagocytic system in mice during 3-methylcholanthrene carcinogenesis.

After an intra-muscular application of 3-methylcholanthrene (MCA) in a dose of 0.2 mg/animal to SPF mice, strain C57B1/10, changes in the lymphocyte nucleoli activation (LNA) and changes of the mononuclear phagocytic system (MPS) in blood and tissues were monitored. The tests were carried out after the first and sixth weeks of the MCA administration, when the tumours were not yet palpable, and 12 weeks after the MCA injection, when the tumours were manifest in 100% of animals. In the first and sixth week after MCA application, the number of non-activable lymphocytes with micronucleoli increased at the expense of the resting ring-shaped forms. In the 12th week, a highly significant drop of active lymphocytes with nucleolonemas and compact nucleoli and particularly a high number of afunctional lymphocytes with micronucleoli were observed in tumour-bearing mice. The phagocytic activity of peripheral mononuclears was significantly increased in the same time interval. MPS activity, monitored on the basis of the colloidal carbon clearance, was stepwise inhibited in animals with originating tumours. It should be pointed out that the LNA test and probably the changes in carbon clearance were early indicators of involvement in chemical carcinogenesis.