Proteomic Profiling of the Blood Serum for Prediction of Premature Delivery.

Mass-spectrometric profiling of the serum in women at weeks 16-17 of gestation was carried out in order to detect proteomic predictors of preterm delivery. Changes in the production of 25 proteins (down-regulation for 13 proteins and up-regulation for 12 proteins) were detected in the sera of women whose pregnancies eventuated in premature deliveries. Among them, proteins with various regulatory functions were distinguished: antioxidant enzymes, chaperons, cytoskeleton proteins, cell adhesion molecules, and proteins involved in angiogenesis, proteolysis, transcription, inflammation processes, binding and transportation of various ligands. These results indicated the formation of proteomic imbalance as early as during trimester II, this eventually leading to premature delivery. The detected serum proteins were suggested as markers for early prediction of premature delivery.

[The imbalance of metal-containing proteins and free metal ions in the amniotic fluid during fetal growth].

The levels of zinc, copper, iron, and magnesium ions, and some of their binding proteins have been investigated in an amniotic fluid under the fetal growth retardation (FGR). FGR, developed under conditions of placental insufficiency, is characterized by a decrease in the content of zinc, iron, and magnesium ions and by an increase in the copper content in the amniotic fluid in the II and III trimesters of pregnancy. During these trimesters the levels of ceruloplasmin, ferritin, and Ca2+,Mg2+-ATPase were lower in FGR, while the level of zinc-a-2-glycoprotein was higher than during the same periods of normal pregnancy. Changes in the parameters studied in the amniotic fluid were associated with developmental disorders of the newborns. These changes obviously have a pathogenetic importance in the development of FGR, and the levels of metal ions and their ratio in the amniotic fluid can be used as markers of the pre- and postnatal pathology.

Use of psychotropic drugs during pregnancy and breast-feeding.

OBJECTIVE: To write clinical guidelines for the use of psychotropic drugs during pregnancy and breast-feeding for daily practice in psychiatry, obstetrics and paediatrics. METHOD: As we wanted a guideline with a high degree of consensus among health professionals treating pregnant women with a psychiatric disease, we asked the Danish Psychiatric Society, the Danish Society of Obstetrics and Gynecology, the Danish Paediatric Society and the Danish Society of Clinical Pharmacology to appoint members for the working group. A comprehensive review of the literature was hereafter conducted. RESULTS: Sertraline and citalopram are first-line treatment among selective serotonin reuptake inhibitor for depression. It is recommended to use lithium for bipolar disorders if an overall assessment finds an indication for mood-stabilizing treatment during pregnancy. Lamotrigine can be used. Valproate and carbamazepin are contraindicated. Olanzapine, risperidone, quetiapine and clozapine can be used for bipolar disorders and schizophrenia. CONCLUSION: It is important that health professionals treating fertile women with a psychiatric disease discuss whether psychotropic drugs are needed during pregnancy and how it has to be administered.

[Imbalance of system of glutamin - glutamic acid in the placenta and amniotic fluid at placental insufficiency].

Metabolism of glutamine and glutamic acid has been investigated in the placenta and amniotic fluid under conditions of placental insufficiency. The development of placental insufficiency is characterized by the increased content of glutamic acid and a decrease of glutamine in both placenta and amniotic fluid. These changes changes were accompanied by changes in the activity of enzymes involved in the metabolism of these amino acids. There was a decrease in glutamate dehydrogenase activity and an increase in glutaminase activity with the simultaneous decrease of glutamine synthetase activity. The compensatory decrease in the activity of glutamine keto acid aminotransferase did not prevent a decrease in the glutamine level. The impairments in the system glutamic acid-glutamine were more pronounced during the development of premature labor.

Diagnostic and remedial capability of transcervical falloposcopy in conjunction with laparoscopy.

STUDY OBJECTIVE: Tubal factor accounts for 25-30% of cases of female infertility. Laparoscopy "Gold Standard" for tubal evaluation. However, it is known that during the initial infection of the fallopian tube mucosal damage occurs, a condition which plays a decisive role in reproduction. MATERIALS AND METHODS: In this prospective randomized study, 468 infertile women with evidence of fallopian tube disease were included. In this, for 256 patients (group 1) after laparoscopic salpingolysis, salpingostomy we performed an additional step operation transcervical falloposcopy tubal dilatation (TFTD). 212 patients (group 2) produced only laparoscopic salpingolysis, salpingostomy. RESULTS: As a result, TFTD patency of the fallopian tubes for coaxial catheter was restored in 50 (78%) of 64 tubes with bilateral total occlusion, in 238 (93%) of 254 with partial occlusion of the bilateral, in 14 (58%) of 24 total unilateral occlusion and 26 (92%) of 28 with partial unilateral occlusion. Total number of pregnancies for one year in the first group of patients was 152 (59.3%), in the second 57 (27.1%), of which in the first group 147 -intrauterine pregnancies (57.4%) and in the second - 46 (21.7%). CONCLUSION: Falloposcopy surgeon provides good opportunities for the diagnosis and treatment of intralumen lesions. Significant increase in the frequency of uterine pregnancy in the case of an additional step - TFTD during salpingolysis, salpingostomy in patients with tubal factor infertility.

[Amino acid composition of amniotic fluid during physiological and complicated pregnancy].

An ion exchange chromatographic technique was used to study the content of 18 free amino acids in the amniotic fluid of women with physiological and complicated pregnancy. The amino acid spectrum of amniotic fluid in placental insufficiency was found to be characterized by the lower content of arginine, serine, glycine, alanine, cysteine, tyrosine, phenylalanine, and glutamine, which are involved in many metabolic processes required to maintain biochemical homeostasis during gestation. There was an opposite tendency for dicarboxylic amino acids and histidine, the levels of which increased, by enhancing nitrogen imbalance. The degree of the found impairments of a number of amino acids considerably increased in case of birth of babies with brain lesions, which permits use of their amniotic fluid determination to predict the neonatal status.

[Proteins-markers of placental insufficiency].

The proteomic analysis of the amniotic fluids of women with physiological pregnancy and pregnancy, complicated with placental insufficiency has been carried out on the II and III trimesters. The following difference in protein patterns have been recognized: i) appearance of several proteins lacking in physiological pregnancy; ii) absence of several proteins detectable during physiological pregnancy--hippocalcin-like protein 1, CDC37-like protein, NKG2D ligand 2 (II trimester), CDC37-like protein, NKG2D ligand 2 (III trimester). The established differences in the amniotic fluid spectrum, obviously, have the pathogenetic meaning in the placental insufficiency development. The revealed proteins of distinction may serve as markers of this obstetrical pathology.

Two mutants of human heparin binding protein (CAP37): toward the understanding of the nature of lipid A/LPS and BPTI binding.

Heparin binding protein (HBP) is an inactive serine protease homologue with important implications in host defense during infections and inflammations. Two mutants of human HBP, [R23S,F25E]HBP and [G175Q]HBP, have been produced to investigate structure-function relationships of residues in the putative lipid A/lipopolysaccharide (LPS) binding site and BPTI (bovine pancreatic trypsin inhibitor) binding site. The X-ray structures have been determined at 1.9 A resolution for [G175Q]HBP and at 2.5 A resolution for the [R23S,F25E]HBP mutant, and the structures have been fully refined to R-factors of 18.2 % and 20.7 %, respectively. The G175Q mutation does not alter the overall structure of the protein, but the ability to bind BPTI has been eliminated, and the mutant mediates only a limited stimulation of the LPS-induced cytokine release from human monocytes. The lipid A/LPS binding property of [G175Q]HBP is comparable with that of native HBP. The R23S,F25E mutations do not affect the binding of lipid A/LPS and BPTI or the LPS-induced cytokine release from human monocytes. This shows that two diverse ligands, lipid A/LPS and BPTI, do not share binding sites. Previously, there was convincing evidence for the proposed lipid A/LPS binding site of HBP. Unexpectedly, the extensive structural changes introduced by mutation of Arg23 and Phe25 do not affect the binding of lipid A/LPS, indicating that another not yet identified site on HBP is involved in the binding of lipid A/LPS.

Heparin-binding protein targeted to mitochondrial compartments protects endothelial cells from apoptosis.

Neutrophil-borne heparin-binding protein (HBP) is a multifunctional protein involved in the progression of inflammation. HBP is stored in neutrophil granules and released upon stimulation of the cells in proximity to endothelial cells. HBP affects endothelial cells in multiple ways; however, the molecular and cellular mechanisms underlying the interaction of HBP with these cells are unknown. Affinity isolation and enzymatic degradation demonstrated that HBP released from human neutrophils binds to endothelial cell-surface proteoglycans, such as syndecans and glypican. Flow cytometry indicated that a significant fraction of proteoglycan-bound HBP is taken up by the endothelial cells, and we used radiolabeled HBP to determine the internalization rate of surface-bound HBP. Confocal and electron microscopy revealed that internalized HBP is targeted to perinuclear compartments of endothelial cells, where it colocalizes with mitochondria. Western blotting of isolated mitochondria from HBP-treated endothelial cells showed that HBP is present in 2 forms - 28 and 22 kDa. Internalized HBP markedly reduced growth factor deprivation-induced caspase-3 activation and protected endothelial cells from apoptosis, suggesting that uptake and intracellular routing of exogenous HBP to mitochondria contributes to the sustained viability of endothelial cells in the context of locally activated neutrophils.

Elimination of the Cys558-Cys566 bond in Lys78-plasminogen--effect on activation and fibrin interaction.

Plasminogen contains a unique disulphide bond, Cys558-Cys566, responsible for the cyclic nature of the peptide sequence surrounding the activation site at Arg561-Val562. A recombinant [Ser558, Ser566]-Lys78-plasminogen variant was produced in which the two cysteine residues were replaced by serine residues. The variant was used to study the functional implications of removing the structural restrains imposed to the activation loop by this disulphide bond. Elimination of the Cys558-Cys566 bond attenuated activation by urokinase-type plasminogen activator (uPA) and tissue-type plasminogen activator (tPA), but resulted in an increased susceptibility to cleavage by trypsin and plasma kallikrein. Two opposite effects on the interaction of plasminogen with streptokinase were produced by modification of this bond; (a) attenuation of the rate at which the active complex with streptokinase was formed and (b) a 7.5-fold increase in plasminogen activation catalysed by this complex. Activation by tPA in the presence of fibrin, in contrast to activation in its absence, was not attenuated by elimination of this disulphide bond. However, the activation rate as a function of plasminogen concentration followed a different saturation curve, and the fibrin degradation pattern was changed. The results suggest that the Cys558-Cys566 disulphide bond is of importance for the specificity of plasminogen. This applies to its activation and also to its role in subsequent fibrin clot degradation.

[Junior physicians educate themselves. Educational value of trainees' work at a department of surgery]. / Reservelaeger uddanner sig selv. Uddannelsesvoerdi af arbejdet på en kirurgisk afdeling.

UNLABELLED: In order to investigate the educational value of trainee surgeons' work, all work done by trainees at a Department of Surgery was recorded in a prospective and consecutive way for four weeks. It was recorded as fixed categories divided into intervals of 15 minutes. The work could be classified in three dimensions: Character, educational value and geography. The youngest trainees performed routine work without educational value. They had some practical jobs which could be done by other personnel. The work did not fulfil the educational claims for a trainee surgeon in phase I. The second group of trainees performed much routine work without educational value. They had no practical jobs that could have been performed by non-doctors. They had some work with educational value but not enough to fulfil the goals of a trainee surgeon in phase II. CONCLUSION: Doctors' clinical education could be improved by giving them work with educational value and supervision. Supervision should be performed by specialists who should also contribute to the routine work without educational value.

[Extracellular DNA in the blood of pregnant women]. / Vnekletochnaia DNK v krovi beremennykh zhenshchin.

The level of extracellular DNA increases in the blood of women during pregnancy. By means of PCR, the full-size Alu repeats were observed among extracellular blood DNA repeats of pregnant women. Furthermore, with Tc65 type primer the PCR method allowed to observe in the blood DNA fragments flanked by inverted Alu repeats (inter Alu repeats). The presence of such a type of inter Alu repeats was estimated in the blood of women being in the first trimester of pregnancy only, but was not estimated among blood DNA fragments of women of the last trimester of pregnancy. It is discussed which types of cells may serve as a source of extracellular blood DNA (either trophoblast cells, lymphocytes, or decidual cells), the significance of such DNA for pregnancy being appreciated.

[The functional activity of the lymphoid cell population in the decidua in the late period of pregnancy]. / Funktsional'naia aktivnost' populiatsii limfoidnykh kletok otpadaiushchei obolochki v pozdnie sroki beremennosti.

Cytotoxic and suppressor activity of decidual cells lymphoid population were analysed in 14 women with normal pregnancy and labour, in 8 women with light pre-eclampsia and 11 women with early and premature rupture of amniotic sac. In late stages of pregnancy decidua is characterised by high suppressor activity of the lymphoid cell population. In early and premature rupture of amniotic sac suppressor activity of decidual cell lymphoid population was significantly decreased.

[The morphofunctional characteristics of the deciduous membrane in pregnancy edema]. / Morfofunktsional'nye osobennosti otpadaiushchei obolochki pri otekakh beremennykh.

An analysis of DNA content in decidua cells and decidua cell number in compact decidual layer was performed in 19 women with physiological pregnancy and labour, in 31 women with oedema of pregnancy, in 29 women with light and in 11 women with severe pre-eclampsia. Significant difference in DNA content in the group with oedema of pregnancy and in the groups with other gestosis forms was demonstrated. Thus, a conclusion is made that oedema may have an independent genesis.

[DNA synthesis and content in human decidual cells at different stages of differentiation based on flow cytometry data]. / Sintez i soderzhanie DNK v kletkakh detsidua cheloveka, nakhodiashchikhsia na raznykh étapakh differentsirovni, po dannym protochnoi tsitometrii.

Flow cytometry analysis of the DNA content of human decidual cells at the physiologically normal pregnancy and in the case of toxicosis was carried out. During the normal pregnancy DNA-histogram parameters were seen to vary: the number of S-phase cells decreased, the coefficient of variation of the G1/0 peak increased. These alterations correlated positively with the increase in the share of decidual cells with properties of terminally differentiated cells. The most pronounced quantitative alterations in variability of DNA content in G1/0 cells were observed in cases of toxicosis of pregnancy. Phenomena of variability of the nuclear DNA in the terminally differentiated decidual cells is considered to be a sign of cell death through apoptosis.

[Several kinetic features of nitrogenase reactions]. / Nekotorye kineticheskie osobennosti nitrogenaznykh reaktsii.

The dependence of the nitrogen reduction rate on N2-concentration under high nitrogen pressures was measured. The reason for the sharp break of the Arrenius curves in nitrogenase reactions at 20 degrees C was examined. The influence of nitrogen on isotope effect catalyzed by nitrogenase was studied. The kinetic schemes that explain the peculiarities of the nitrogenase reactions were analyzed by the graph's theory. On the base of new conception of inhibitor-"interceptor" of electrons the literature data were discussed.