Post-partum evaluation of amylin in lean patients with gestational diabetes mellitus.

The aim of our study was to compare the secretion of amylin, as well as glucose, insulin and C-peptide at baseline and in response to glucagon stimulation in 26 lean women with gestational diabetes mellitus (GDM) and in 19 age- and BMI-matched pregnant women with normal glucose tolerance (NGT). Intravenous 1-mg glucagon stimulation test was performed 6 weeks after delivery. Fasting and stimulated glucose levels were significantly higher in GDM patients than in subjects with NGT ( p<0.01 at 0 and 6 min; glucose area under the curve (AUC), 604.8+/-41.8 mg/6 min vs. 572.4+/-52.4 mg/6 min, p<0.05). Insulin AUC was also markedly higher in GDM subjects than in healthy controls (373.9+/-144.2 micro IU/6 min vs. 283.7+/-139.1 micro IU/6 min, p<0.05). There was no difference in fasting C-peptide levels between the groups studied, but stimulated concentrations, as well as C-peptide AUC were significantly higher in patients with GDM ( p<0.01 at 1 min and p<0.005 at 6 min; AUC, 27.4+/-11.3 pmol/6 min vs. 18.4+/-6.9 pmol/6 min, p<0.01). Amylin levels were higher in GDM group in comparison to healthy subjects ( p<0.005 at 1 and 6 min; amylin AUC, 113.3+/-51.2 pg/6 min vs. 72.5+/-15.7 pg/6 min; p=0.14), but in contrast to the other hormones, did not rise in response to glucagon injection. In conclusion, our results provide evidence that in patients with GDM in the post-partum period, the levels of amylin, as well as the secretion of insulin and C-peptide remain elevated, when compared to women with NTG. Further investigations are needed to clarify the significance of this elevation as a predictive factor for the development of late maternal type 2 diabetes.

[Evaluation of lipid peroxidation and acid-base status in cord blood of newborns after diabetes in pregnancy]. / Ocena peroksydacji lipidów i równowaga kwasowo-zasadowa u noworodka z ciazy powiklanej cukrzyca.

The purpose of our study was to evaluate lipid peroxidation products and scavenging enzyme activity in placenta and cord blood as well as the estimation of acid-base status and blood gases. Seventy five pregnant patients and their newborns were investigated. Twenty eight had pre-gestational diabetes mellitus (PGDM) and 19 gestational diabetes mellitus (GDM). The following parameters were measured: malondialdehyde (MDA) concentrations, glutathione (GSH) levels, the activity of CuZn dismutase (SOD) (Bioxytech, France). Base excess, pO2, pCO2 and pH were measured in arterial and venous samples. Statistical analysis was performed using Mann-Whitney U test. MDA levels and GSH content increased significantly, while SOD activities declined in diabetic group. Newborns of PDGM mothers had essentially diminished pH and rised both, pCO2 and base deficit. There were no any significant differences in parameters of acid-base balance in newborns of patients with GDM as compared with healthy patients. Our results suggest, that in diabetic patients the fetuses are exposed to increased oxidative stress. The evaluation of antioxidant defence and lipid peroxidation, apart from routine measurement of acid-base balance, might serve as a useful marker of fetal distress in diabetic patients.

Lipid peroxidation, antioxidant defence and acid-base status in cord blood at birth: the influence of diabetes.

Pregnancy complicated by poor control of diabetes is associated with a higher risk of embryopathies, spontaneous abortions and perinatal mortality. A number of authors suggest an involvement of reactive oxygen species (ROS) in diabetic pregnancy. Determining lipid peroxidation products (LP), scavenging enzyme activities and the umbilical cord blood's acid-base balance may contribute to an adequate diagnosis of the neonate at birth. Nevertheless, such measurements seem to have limited value in practical clinical routine. The present study evaluates LP, antioxidant defence and acid-base status related to diabetic pregnancy. Twenty-eight women with type 1 diabetes (PGDM), 19 with gestational diabetes (GDM) and 13 control cases were investigated. An additional control group consisted of 15 healthy patients with negative diabetic history; all women underwent vaginal delivery. Immediately after delivery cord blood samples and placental tissue were collected for malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) determination. Additionally, pH, pCO2, pO2 and base excess were measured in both vessels and compared to identify and exclude double venous samples. MDA levels in both cord blood and placental homogenates were significantly higher in both pregestational and gestational diabetic groups, but SOD activity was significantly diminished. Cord blood GSH was markedly elevated in PGDM and GDM. We have also shown significant differences in acid-base parameters in infants of PGDM group. Statistical analysis was performed using the Mann-Whitney U-test. These findings indicate an excessive oxidative stress in pregnancy complicated by diabetes mellitus. Evaluating LP products and scavenging enzyme activities may be valuable, sensitive indexes of fetal/neonatal threat in diabetic pregnancy in humans. Since oxidative stress is an important pathway for fetal injury, we believe that obtaining adequate measurements at the time of birth would contribute to clarifying the fetal/neonatal status in a medical and legal context and might be of value in altering therapy in newborn infants.

[Diabetic patient and reproduction].

Current views and opinions on pregnancy in diabetic mothers are presented. Special attention is paid to carbohydrate metabolism, pregestational diabetes mellitus (PGDM), gestational diabetes mellitus (GDM) and unclear heterogenous etiology of gestational diabetes mellitus (GDM). Suggestions for identification, diagnosis and treatment of GDM with an emphasis on early screening strategy are given. Obstetrical management and possible perinatal complications are discussed.

[Strategy in gestational diabetes]. / Strategia postepowania w cukrzycy ciezarnych.

This paper reviews current literature on strategic policy in gestational diabetes (GDM). Emphasis is placed on early identification of carbohydrate intolerance. Definitive guidelines related to screening procedures, diagnostic criteria, glucose monitoring with threshold for insulin therapy in GDM were briefly described. Pregnancy termination and delivery management have been discussed. The importance of postpartum follow-up is pointed out. Special attention is paid to the association with increased antenatal risk of subsequent diabetes and consequently raised perinatal morbidity and/or mortality. For appropriate health-care a cooperation between patients, obstetricians and diabetologists is suggested.

[Analysis of selected methods for intrauterine stimulation of fetal pulmonary maturation]. / Analiza wybranych metod wewnatrzmacicznej stymulacji dojrzewania ukladu oddechowego u plodu.

Preterm delivery is still a major cause of infant morbidity and mortality. Premature infants develop numerous complications including respiratory distress syndrome, intraventricular hemorrhage, patent ductus arteriosus, necrotising enterocolitis, etc. Respiratory distress syndrome is the leading problem in perinatology. The incidence of RDS accounts for 70% of all infants delivered before 30 week's gestation and about 20% neonates born between 32-37 week of pregnancy. Our paper presents current opinions and is a critical review of relevant literature on antenatal treatment before anticipated preterm deliveries. Recently performed meta-analysis prove that glucocorticoid therapy is effective in reducing aforementioned pathologies of prematurity. After brief account on effects of several hormones on fetal pulmonary maturity some beneficial interactions for enhanced fetal maturity are pointed out. Glucocorticoid mechanism of action in target cells and its biochemical implications are reviewed. The use of antenatal corticoids for fetal maturation and possible adverse maternal effects follow. Finally both short-term and long-term benefits of ANS are discussed. Thus there are convincing data to support the use of ANS in fetal pulmonary maturation. The current findings suggest that the improvement of respiratory outcome may depend on enhanced expression of phospholipids and proteins of the surfactant system and enzymes of antioxidant systems. We believe that use of antenatal corticosteroids for fetal maturation results in improvement of neonatal outcome and yields substantial savings in health care costs.

[Intrauterine stimulation for fetal respiratory system maturation; benefits and risks]. / Wewnatrzmaciczna stymulacja dojrzewania ukladu oddechowego; korzysci a ryzyko.

The use of perinatal steroid therapy, first introduced in 1972 is effective in precocious maturation of human lungs. Antenatal corticosteroid therapy results in reduction of fetal mortality, respiratory distress syndrome, intraventricular hemorrhage in preterm babies. These benefits extend to a broad range of gestational age. They comprise the interval between 24 and 34 weeks of human pregnancy and are not limited by the infant's gender or race. The beneficial effects of corticosteroids are the best pronounced after more than 24 hours from the beginning of the treatment. Noteworthy is that therapy less than 24 hours of duration may also improve outcomes. In the presence of premature rupture of membranes, or better with intact membranes, antenatal corticosteroids reduce frequency of RDS, IVH and finally mortality and morbidity. Review of meta-analyses based on randomized trials supports general option that premature infants whose mothers received corticosteroids before delivery are less likely to develop RDS and its complications. Recent data showed that benefits derived from ANS are additive to those of surfactant therapy, rendering the latter more effective. Followup of children up to 12 years of age indicate that ANS do not impair physical growth or psychomotor development. Short-term adverse effects including maternal infection, maternal pulmonary edema were not clearly demonstrated. Pulmonary edema has not been reported when ANS were used alone (i.e. not in combination with betamimetic tocolytics). No long-term unwanted effects on maternal adrenal function have been observed. There is no serious maternal risk resulting from immunosuppressive effect of corticosteroid therapy on maternal immune system. Although glucocorticoid therapy is likely to provoke insulin resistance, and thereby deterioration in diabetic control, and potentially causes cortisol resistance in the fetal lung, the results of scarce randomized trials are not conclusive. In any rate strict control of maternal diabetes mellitus reduces incidence of RDS. Current available data are not indicative of higher risk of fetal mortality in association with maternal hypertensive disease and ANS. In conclusion, most randomized trials of ANS has provided a positive evidence of efficacy and safety of this highly cost effective therapy in most common clinical situations. However, further trials and more precise estimates are justified on ANS treatment specifically related to blood glucose control and evidence concerning the promotion of fetal lung maturity in babies of women with diabetes mellitus. Although benefits of the corticosteroid therapy are beyond any doubts, more experience is needed to assess the effect of ANS on maternal and/or fetal infection in presence of premature rupture of membranes. And finally, further assessments are required on antenatal corticosteroids with dose regimens in patients with multifetal gestation, more common after wide use of techniques of the assisted human reproduction.

[Use of oxygen therapy during labour and acid-base status in the newborn]. / Tlenoterapia stosowana w czasie porodu a parametry równowagi kwasowo-zasadowej u noworodka.

The effect of supplemental maternal oxygen therapy on acid-base status has for years been a subject of controversies. There is still no general agreement about oxygen administration in pregnancy or in labour. Many clinicians believe that in normal pregnancy maternal oxygen treatment has merely negligible, if any, influence on the foetus. Maternal oxygen therapy has frequently been suggested when oxygen supply to the foetus was insufficient, viz. in antepartum or intrapartum hypoxia. The aim of this study was to determine whether a brief maternal hyperoxygenation during caesarean section or during the second stage of normal delivery affects cord blood acid-base status measured at birth. Patients with uneventful term pregnancy admitted to the delivery room in spontaneous labour or indicated for elective caesarean section were matched and prospectively ascribed to the control or treatment group. The latter received 60% oxygen for ca. 10 min at 15 L/min by a face mask. We studied 41 normal term infants of healthy mothers (24 from caesarean sections, the remaining 17 delivered vaginally). The control group consisted of normal newborns chosen according to the same criteria except that their mothers had no oxygen supplementation. Immediately after delivery, umbilical cord paired blood samples were drawn. Arterial and venous specimens were analysed for blood gases and acid-base balance parameters. Statistical analysis assisted by computer software was performed using Mann-Whitney U test. A p value < 0.05 was considered significant. All tested parameters related to the acid-base status and blood gases were similar in the treatment and control groups. Concluding, a short period of maternal hyperoxygenation either in the second stage of vaginal spontaneous delivery or during caesarean section did not affect umbilical cord blood acid-base status measured at birth.

Lipid peroxidation and scavenging enzyme activity in streptozotocin-induced diabetes.

The aim of this study was to evaluate lipid peroxidation and scavenging enzyme activity in streptozotocin-induced diabetes, and then to establish whether moderate doses of nonenzymatic antioxidant vitamin E play a role in the antioxidant defence system in diabetic pregnant rats and their offspring. The study group consisted of 30 normal female Wistar rats, which were given a single dose of streptozotocin (40 mg/kg) and were mated 7 days later. Subsequently, the diabetic animals were divided into two matched groups: the first supplemented with vitamin E (30 mg/100 g chow), and the other fed with a standard diet lacking vitamin E. Controls consisted of 15 pregnant rats. On the first day after delivery, the rats were decapitated and homogenates of maternal liver and uterus as well as neonatal lungs and liver were prepared. Then the following parameters were measured: malondialdehyde (MDA) concentrations in the homogenates and blood serum, glutathione (GSH) levels, the activity of CuZn-superoxide dismutase (SOD) and glutathione peroxidase (GPx), and glycaemia. The neonates of diabetic rats were smaller than the healthy ones and serum glucose concentration was markedly higher in the diabetic animals. MDA levels were significantly increased, whereas GSH, SOD and GPx were markedly diminished in the diabetic adult rats and their offspring in comparison to the control group. In the animals supplemented with alpha-tocopherol, MDA concentrations were significantly lower, GSH content and SOD activities were markedly elevated most tissues studied, whereas GPx remained unchanged. We conclude that, by monitoring the activity of selected scavenging enzymes, information on ongoing biological oxidative stress and thereby on the fetus/neonate status may be obtained. Our results suggest that diabetic pregnant rats and their neonates are exposed to an increased oxidative stress and that vitamin E supplementation may reduce its detrimental effects.

Antioxidant therapy and streptozotocin-induced diabetes in pregnant rats.

The aim of our study was to analyse the effect of chronic hyperglycaemia on lipid peroxidation and scavenging enzyme activity in pregnant animals and their offspring supplemented and not supplemented with vitamin E - a natural antioxidant. Thirty pregnant female Wistar rats were used in our experiments. Diabetes was induced on day 7 of pregnancy using a single does of streptozotocin (40 mg/kg). Diabetic animals were divided into two equal groups: vitamin E supplemented and those fed with standard diet. Our controls consisted of 15 healthy rats. On day 1 after delivery homogenates of maternal liver and uterus as well as neonatal lungs and liver were prepared. Then the following parameters were measured: malondialdehyde (MDA) concentrations in the homogenates and blood serum, glutathione (GSH) levels, the activity of CuZn superoxide dismutase (SOD) and glutathione peroxidase (GPx) (Bioxytech, France). Statistical analysis was performed using Mann-Withney U test. The neonates of diabetic rats were smaller than those from healthy rats and serum glucose concentration was markedly higher in diabetic animals, both in mothers and neonates. MDA levels increased significantly, whereas GSH content and SOD as well as GPx activities were markedly diminished in diabetic pregnant rats and their offspring in comparison with the control group. In animals supplemented with tocopherol, MDA concentrations declined significantly, GSH contents and SOD activities were markedly elevated in almost all types of tissues studied, whereas glutathione peroxidase remained suppressed. Our results suggest that diabetic pregnant rats and their neonates are exposed to oxidative stress (OS), but vitamin E supplementation could in part reduce the imbalance between uncontrolled reactive oxygen species generation and scavenging enzyme activity, and may potentially serve as a useful prophylactic factor against OS development:

[Familial translocation t (9;16) in a patient with irregular menstrual cycles]. / Rodzinna translokacja t (9;16) u pacjentki z nieregularnymi cyklami miesiaczkowymi.

In this report we present a family with identified carriers of unique reciprocal translocation t (9; 16) (q31; q13) detected through karyotyping of the patient with irregularity of menstrual cycles. Genetic risk for birth of a child with congenital anomalies was estimated as low (0.6%). However, risk for abortion was high. We suggest introducing cytogenetic studies in such cases.

[Gene therapy perspectives in cystic fibrosis]. / Perspektywy zastosowania terapii genowej w mukowiscydozie.

Cystic fibrosis (CF) is a frequent autosomal recessive genetic disease. The isolation of the gene at the CF locus assigned to the long arm of chromosome 7 band q 31 and defining description of its protein named CFTR (cystic fibrosis transmembrane conductance regulator) promoted understanding the basic biochemical defect. Brief review of relevant literature demonstrates that glycoprotein CFTR is a chloride channel and is activated by a combination of phosphorylation by protein kinase A and binding of ATP. Most common mutation of CF gene, a deletion of the three nucleotides encoding phenylalanine (Delta F508) results in disturbance of chloride transport through membrane of epithelial cells involved in pathomechanism of CF. The way for gene therapy in CF is open, however therapeutic progress is noted on both pharmacologic arena and on the gene cure front. Recombinant vectors utilizing the adenovirus system with high efficiency of CFTR gene transfer to airway epithelium demonstrated in a rat model look promising. The use of retroviruses for CFTR transfer is also advanced mode of somatic gene therapy. An alternative approach suggesting the use of germ line cells is prerequisite of the development of the preimplantation/preconception genetic CF diagnosis. A number of safety and efficacy issues have to be addressed for all approaches before human trials can be implemented.

Cord blood triglyceridemia in cases of placental insufficiency.

Cord blood triglyceride concentration increased in cases of placental insufficiency. The significant relationship has been found between elevated triglyceride, decreased Apgar score, and low pH values. We suggest that the measurement of cord blood triglyceridemia might be used as a valuable indicator of chronic fetal distress.

[Degradation and consumption of one's own glycogen in placental tissue in pregnancy complicated by gestosis and anemia]. / Rozklad i zuzycie glikogenu wlasnego przez tkanke lozyska z ciazy powiklanej gestoza i niedokrwistoscia.

We estimated in vitro degradation and consumption of glycogen by placental tissue derived from pregnancies complicated by gestosis and anemia. Placental tissue was incubated in suitable medium. Glycogen concentration either before or after incubation was assayed. Glycogen consumption was expressed as micromoles of glucose per 1 g of wet tissue, the percentage of glycogen consumption has been shown in tables. It is concluded that in advanced gestosis and severe anemia of pregnancy the consumption of native glycogen is increased, consequently it may lead to metabolic insufficiency of human placenta.

[Effect of diseases in pregnancy on certain placental functions. Enzymatic systems degrading placental glycogen in pregnancy with gestosis]. / Ocena wplywu chorób ciezarnej na wybrane funkcje lozyska. Uklady enzymatyczne rozkladajace glikogen w lozysku od Ciezarnej z gestoza.

Pathways of human placental glycogen degradation either in normal or gestotic pregnancy were examined. Determinations of glycogen phosphorylase served as an index of glycogen cleavage in the phosphorylitic pathway. The activity of hydrolytic route was measured by estimating placental glucoamylase. Samples of placental tissue were obtained after delivery (between the 36th and the 40th week of pregnancy). In placentas derived from gestotic cases elevated activity of phosphorylase A, the decrease in phosphorylase B activity and the rise in glucoamylase were found. It suggests that gestosis may evoke certain impairment of placental glycogen metabolism deteriorating chiefly the glycogen degradation as it may be observed in hypoxic experiments.

Umbilical cord gamma-glutamyl transpeptidase and placental dysfunction.

Placental dysfunction has been suspected if human placental lactogen level and/or cystine-aminopeptidase activity were lower than 10th centile. Significant rise in gamma-glutamyl transpeptidase (GGTP) has been found in samples of arterial cord blood of newborns born to mothers with placental insufficiency. We observed the relationship between GGTP activity and decrease in cord pH and lower Apgar score of corresponding infants. We suggest that chronic hypoxemia as a consequence of placental dysfunction may result in damage of fetal liver cells.

Cloning of human lysozyme gene and expression in the yeast Saccharomyces cerevisiae.

cDNA clones encoding human lysozyme were isolated from a human histiocytic cell line (U-937) and a human placenta cDNA library. The clones, ranging in size from 0.5 to 0.75 kb, were identified by direct hybridization with synthetic oligodeoxynucleotides. The nucleotide sequence coding for the entire protein was determined. The derived amino acid sequence has 100% homology with the published amino acid (aa) sequence; the leader sequence codes for 18 aa. Expression and secretion of human lysozyme in Saccharomyces cerevisiae was achieved by placing the cloned cDNA under the control of a yeast gene promoter (ADH1) and the alpha-factor peptide leader sequence.

Two zinc fingers of a yeast regulatory protein shown by genetic evidence to be essential for its function.

The best-understood protein structure involved in DNA binding is the helix-turn-helix motif. A second DNA-binding domain, the finger structure, has been proposed on the basis of sequence analysis, partial proteolysis and zinc content of Xenopus transcription factor TFIIIA. Other eukaryotic proteins were subsequently found to contain contiguous repeat units of the postulated finger motif. Each repeat unit contains thirty amino acids and is thought to bind a zinc atom using two cysteines and two histidines as ligands. The protein loop or finger between apparent zinc ligands is rich in DNA-binding residues and is thought to make specific contacts with DNA. The yeast protein ADR1, a positive regulator of transcription of the gene ADH2, contains two finger domains in a region of the protein required for transcriptional activation. Nineteen independently isolated adr1 mutations induced by hydroxylamine were found at nine different amino-acid positions, seven of which are in the two finger domains. All four mutations that altered invariant cysteine or histidine residues led to an adr1 null phenotype. Only one other mutation caused an adr1 null phenotype. Thus, one finger domain is not sufficient for ADR1 activity. This provides the first evidence that, as is consistent with the proposed model, the invariant cysteine and histidine residues are essential for the formation of the finger structure.

Cord blood serum creatine kinase isoenzymes with placental dysfunction.

It has been suggested that perinatal asphyxia is not generally followed by neurological impairment unless there is preexisting chronic fetal distress. In cases of brain damage one can observe elevated levels of CK-BB. The purpose of our study was to evaluate CK isoenzymes in umbilical cord blood sera of newborns affected by chronic fetal distress. Fetal distress reflected by placental dysfunction was characterized by a diminished HPL level and decreased activity of CAP. We estimated CK isoenzymes with the use of DEAE-sepharose CL-6B column chromatography. Total CK activity was measured using kits supplied by Boehringer-Mannheim (Monotest CK-NAC aktiviert). The clinical state of examined newborns was estimated. Investigations were carried out in the group of 57 infants delivered after 37 weeks of gestation. Total CK activity in cord sera ranged from 40 to 400 U/l. Our results showed a significant rise of CK-BB activity in cord sera of newborns delivered from pregnancies with placental dysfunction (figure 2) as well as in cases of asphyxiated infants (figure 3). We were unable to demonstrate differences in total CK, CK-MM and CK-MB activities in all examined groups of newborns. Other authors have confirmed that severe asphyxia results in increase in CK-BB activity in cord blood. Infants with ominous fetal heart rate patterns have higher CK-BB activity. There are several possible sources for CK-BB activity in umbilical cord blood sera, i.e. fetal brain, lung, gastrointestinal tract, placenta and uterus. It appears that the brain is most likely the source of elevated CK-BB activity found in cord blood in cases of placental dysfunction.