Marcadores moleculares de la asfixia perinatal / Molecular markers of perinatal asphyxia

Introducción: la asfixia perinatal es un problema de salud que puede acarrear alteraciones del neurodesarrollo en los recién nacidos. Las determinaciones en suero de enolasa específica de neurona, lactato deshidrogenasa y aspartato amino transferasa han sido utilizadas como marcadores de asfixia perinatal. Objetivos: evaluar el valor de las determinaciones en suero de lactato deshidrogenasa, aspartato amino transferasa y enolasa específica de neurona como marcadores moleculares de la asfixia perinatal. Métodos: se realizó un estudio observacional descriptivo de corte transversal. Se trabajó con una muestra intencional de 41 recién nacidos asfícticos, clasificados con distintos grados de encefalopatía hipóxico-isquémica según los criterios de Sarnat. Se tomaron muestras de suero al momento del nacimiento y a las 72 horas siguientes. Las determinaciones en suero de enolasa específica de neurona se realizaron por ELISA. Se cuantificó lactato deshidrogenasa y aspartato amino transferasa por espectrofotometría. Resultados: todos los pacientes presentaron valores elevados en suero, de los tres analitos, a las 24 y 72 horas de nacidos. Los valores enzimáticos no variaron significativamente entre las 24 y 72 horas de nacidos sin tomar en cuenta el grado de encefalopatía hipóxico-isquémica. Existe correlación positiva entre los valores enzimáticos a las 24 y a las 72 horas de enolasa específica de neurona y lactato deshidrogenasa. No fue posible diferenciar el grado de encefalopatía hipóxico-isquémica a través de los niveles en suero de estas enzimas. Conclusiones: los valores de estas determinaciones enzimáticas contribuyen a describir desde el punto de vista bioquímico el cuadro del neonato con asfixia perinatal(AU)

Introduction: perinatal asphyxia is a health problem which may cause neurodevelopmental alterations in newborns. Serum determinations of neuron-specific enolase, lactate dehydrogenase, and aspartate aminotransferase have been used as markers of perinatal asphyxia. Objectives: evaluate the value of serum determinations of lactate dehydrogenase, aspartate aminotransferase and neuron-specific enolase as molecular markers of perinatal asphyxia. Methods: a cross-sectional observational descriptive study was conducted of 41 asphyxiated newborns classified as different grades of hypoxic-ischemic encephalopathy according to Sarnat's scale. Serum samples were taken at birth and 72 hours later. Serum determinations of neuron-specific enolase were obtained by ELISA. Lactate dehydrogenase and aspartate aminotransferase were quantified by espectrophotometry. Results: all the patients had high serum values of the three analytes 24 and 72 hours after birth. Enzyme values did not vary significantly from 24 to 72 hours after birth, not considering the grade of hypoxic-ischemic encephalopathy. A positive correlation was found between enzyme values for neuron-specific enolase and lactate dehydrogenase at 24 and 72 hours. It was not possible to differentiate the grade of hypoxic-ischemic encephalopathy via the serum levels of these enzymes. Conclusions: the values of these enzyme determinations contribute to describe the status of neonates with perinatal asphyxia from a biochemical point of view(AU)

Perinatal asphyxia leads to PARP-1 overactivity, p65 translocation, IL-1ß and TNF-α overexpression, and apoptotic-like cell death in mesencephalon of neonatal rats: prevention by systemic neonatal nicotinamide administration.

Perinatal asphyxia (PA) is a leading cause of neuronal damage in newborns, resulting in long-term neurological and cognitive deficits, in part due to impairment of mesostriatal and mesolimbic neurocircuitries. The insult can be as severe as to menace the integrity of the genome, triggering the overactivation of sentinel proteins, including poly (ADP-ribose) polymerase-1 (PARP-1). PARP-1 overactivation implies increased energy demands, worsening the metabolic failure and depleting further NAD(+) availability. Using a global PA rat model, we report here evidence that hypoxia increases PARP-1 activity, triggering a signalling cascade leading to nuclear translocation of the NF-κB subunit p65, modulating the expression of IL-1ß and TNF-α, pro-inflammatory molecules, increasing apoptotic-like cell death in mesencephalon of neonate rats, monitored with Western blots, qPCR, TUNEL and ELISA. PARP-1 activity increased immediately after PA, reaching a maximum 1-8 h after the insult, while activation of the NF-κB signalling pathway was observed 8 h after the insult, with a >twofold increase of p65 nuclear translocation. IL-1ß and TNF-α mRNA levels were increased 24 h after the insult, together with a >twofold increase in apoptotic-like cell death. A single dose of the PARP-1 inhibitor nicotinamide (0.8 mmol/kg, i.p.), 1 h post delivery, prevented the effect of PA on PARP-1 activity, p65 translocation, pro-inflammatory cytokine expression and apoptotic-like cell death. The present study demonstrates that PA leads to PARP-1 overactivation, increasing the expression of pro-inflammatory cytokines and cell death in mesencephalon, effects prevented by systemic neonatal nicotinamide administration, supporting the idea that PARP-1 inhibition represents a therapeutic target against the effects of PA.

Thioredoxin 1 and glutaredoxin 2 contribute to maintain the phenotype and integrity of neurons following perinatal asphyxia.

BACKGROUND: Thioredoxin (Trx) family proteins are crucial mediators of cell functions via regulation of the thiol redox state of various key proteins and the levels of the intracellular second messenger hydrogen peroxide. Their expression, localization and functions are altered in various pathologies. Here, we have analyzed the impact of Trx family proteins in neuronal development and recovery, following hypoxia/ischemia and reperfusion. METHODS: We have analyzed the regulation and potential functions of Trx family proteins during hypoxia/ischemia and reoxygenation of the developing brain in both an animal and a cellular model of perinatal asphyxia. We have analyzed the distribution of 14 Trx family and related proteins in the cerebellum, striatum, and hippocampus, three areas of the rat brain that are especially susceptible to hypoxia. Using SH-SY5Y cells subjected to hypoxia and reoxygenation, we have analyzed the functions of some redoxins suggested by the animal experiment. RESULTS AND CONCLUSIONS: We have described/discovered a complex, cell-type and tissue-specific expression pattern following the hypoxia/ischemia and reoxygenation. Particularly, Grx2 and Trx1 showed distinct changes during tissue recovery following hypoxia/ischemia and reoxygenation. Silencing of these proteins in SH-SY5Y cells subjected to hypoxia-reoxygenation confirmed that these proteins are required to maintain the normal neuronal phenotype. GENERAL SIGNIFICANCE: These findings demonstrate the significance of redox signaling in cellular pathways. Grx2 and Trx1 contribute significantly to neuronal integrity and could be clinically relevant in neuronal damage following perinatal asphyxia and other neuronal disorders.

Magnesium sulfate effect on erythrocyte membranes of asphyxiated newborns.

OBJECTIVES: Magnesium sulfate has been recognized as a neuroprotective agent against hypoxia-ischemia, mainly by the protection from the excitotoxicity associated with increased glutamate concentration. However, the mechanism of MgSO4 action is not fully understood and is considerably controversial. DESIGN AND METHODS: During the 2 first hours of life, the asphyxiated full-term newborns were treated intravenously with one dose of MgSO4 250 mg/kg body weight. At birth, after 6 and 48 h of life the activity of ATP-dependent enzymes in erythrocyte membranes: Mg2+-ATPase, Ca2+-ATPase, protein kinases A and C, were determined. Using monoclonal antibodies, the band 3 and its phosphotyrosine level were also assayed. RESULTS: The time-dependent decrease of Ca2+-ATPase activity was detected in untreated newborns, whereas MgSO4 prevented this reduction. After 48 h, protein kinases activities differed in MgSO4-treated and untreated groups. Magnesium therapy increased the amount of band 3 and diminished proteolytic degradation of this protein. CONCLUSION: Our results demonstrated, for the first time, that magnesium sulfate treatment significantly altered the activities of some important enzymes in erythrocyte membrane from asphyxiated newborns. It also reduced the post-asphyxial damages of membrane compounds. These data may partly explain the molecular mechanisms of MgSO4 action in asphyxiated newborns.

Hypoxia-induced modification of poly (ADP-ribose) polymerase and dna polymerase beta activity in cerebral cortical nuclei of newborn piglets: role of nitric oxide.

Previous studies have shown that poly (ADP-ribose) polymerase (PARP) and DNA polymerase beta, nuclear enzymes, are associated with cell replication and DNA repair. The present study tests the hypothesis that hypoxia results in increased PARP and DNA polymerase activity in cerebral cortical neuronal nuclei to repair the hypoxia-induced damage to genomic DNA. Studies were conducted in 13 anesthetized and ventilated newborn piglets (age 3-5 days) divided into normoxic (n=5) and hypoxic (n=8) groups. Hypoxia was induced by decreasing inspired oxygen from 21% to 7% for 60 min. Cerebral tissue hypoxia was documented biochemically by determining the tissue levels of ATP and phosphocreatine (PCr). Following isolation of the cortical neuronal nuclei, the activity of PARP and DNA polymerase beta was determined. During hypoxia, the tissue ATP level decreased by 73% from 4.12+/-0.67 micromol/g brain to 1.12+/-0.34 micromol/g brain, and PCr decreased by 78% from 4.14+/-0.68-0.90+/-0.20 micromol/g brain. In hypoxic neuronal nuclei, PARP activity significantly increased from 5.88+/-0.51 pmol NAD/mg protein/h in normoxic nuclei to 10.04+/-2.02 (P=0.001). PARP activity inversely correlated with tissue ATP (r=0.78) and PCr levels (r=0.81). Administration of N-nitro-L-arginine prior to hypoxia decreased the hypoxia-induced increase in PARP activity by 67%. Endogenous DNA polymerase beta activity increased from 0.96+/-0.13 in normoxic nuclei to 1.39+/-0.18 nmol/mg protein/h in hypoxic nuclei (P<0.005). DNA polymerase beta activity in the presence of exogenous template increased from 1.54+/-0.14 in the normoxic to 2.42+/-0.26 nmol/mg protein/h in the hypoxic group (P<0.005). DNA polymerase beta activity in the presence or absence of template inversely correlated with the tissue ATP (r=0.95 and 0.84, respectively) and PCr levels (r=0.93 and 0.77, respectively). These results demonstrate that the activity of PARP and DNA polymerase beta enzymes increase with the increase in degree of cerebral tissue hypoxia. Furthermore, the results demonstrate a direct correlation between the PARP and the DNA polymerase beta activity. We conclude that tissue hypoxia results in increased PARP and DNA polymerase beta activities indicating activation of DNA repair mechanisms that may result in potential neuronal recovery following hypoxia and the hypoxia-induced increase in PARP activity is NO-mediated.

Catecholamine-induced oligodendrocyte cell death in culture is developmentally regulated and involves free radical generation and differential activation of caspase-3.

Oligodendrocyte cultures were used to study the toxic effects of catecholamines. Our results showed that catecholamine-induced toxicity was dependent on the dose of dopamine or norepinephrine used and on the developmental stage of the cultures, with oligodendrocyte progenitors being more vulnerable. A role for oxidative stress and apoptosis on the mechanism of action of catecholamines on oligodendrocyte cell death was next assessed. Catecholamines caused a reduction in intracellular glutathione levels, an accumulation in reactive oxygen species and in heme oxygenase-1, the 32 kDa stress-induced protein. All these changes were prevented by N-acetyl-L-cysteine, a thiocompound with antioxidant activity and a precursor of glutathione, and were more pronounced in progenitors than mature cells, which could contribute to their higher susceptibility. Apoptotic cell death, as assessed by activation of caspase-9 and -3 and cleavage of poly(ADP-ribose) polymerase (a substrate of caspase-3), was only observed in oligodendrocyte progenitors. Pretreatment with zVAD, a general caspase inhibitor, prevented activation of caspase-9 and -3, DNA fragmentation, and decreased progenitors cell death. Furthermore, the expression levels of procaspase-3 and the ratio of the proapoptotic protein bax to antiapoptotic protein bcl-xl were several folds higher in immature than mature oligodendrocytes. Taken together, these results strongly suggest that the catecholamine-induced cytotoxicity in oligodendrocytes is developmentally regulated, mediated by oxidative stress, and have characteristics of apoptosis in progenitor cells.

Protein kinases activities in erythrocyte membranes of asphyxiated newborns.

OBJECTIVES: Perinatal asphyxia represents a major cause of acute brain impairment and mortality in neonates. To develop the effective therapies able to reduce post-asphyxial damages, the understanding of biochemical processes accompanying asphyxia appears to be of the great relevance. DESIGN AND METHODS: The activities of protein kinases A and C, and tyrosine kinases in erythrocyte membranes of healthy and asphyxiated neonatals were compared. Using monoclonal antibodies the band 3 presence and its phosphotyrosine levels were assayed. RESULTS: In asphyxiated erythrocyte membranes the activities of PKA and tyrosine kinases increased, whereas the activity of PKC was reduced in relation to healthy newborns. Under asphyxia the band 3 has been overphosphorylated; however, its amount decreased. CONCLUSION: These findings may provide some evidence for a potential role of asphyxia in disturbance of phosphorylation processes in erythrocytes, as reflected by altered protein kinases activities. The diminished band 3 presence may be partially responsible for the impairment of erythrocyte function.

Effects of hypothermia on neonatal hypoxic-ischemic brain injury in the rat: phosphorylation of Akt, activation of caspase-3-like protease.

Neuroprotective mechanisms of hypothermia have not been clearly established especially in the immature brain. To investigate the effect of hypothermia on cell death and cell survival signal pathways, we studied caspase-3-like activity and activation of Akt in a rat model of neonatal hypoxic-ischemic (H-I) brain injury. Seven-day-old rats underwent a combination of left common carotid artery ligation and exposure to 8% O(2) for 1-h (n=32). During recovery, the body temperature was reduced to 30 degrees C for 24 h in 16 animals, but was kept at 37 degrees C in 16 animals. Post-ischemic hypothermia was shown to diminish the caspase-3-like activity compared to normothermia at 6 and 24 h after H-I. Phospho-Akt was increased during the early reperfusion period after H-I in the normothermia group, but hypothermia rather decreased this enhanced phosphorylation of Akt following H-I. These results indicated that hypothermia may have some depressant effects on both cell death and cell survival signal pathways, and that Akt conceivably may not play a major role in the neuroprotective effect of hypothermia in the immature brain.

Intracellular generation of free radicals and modifications of detoxifying enzymes in cultured neurons from the developing rat forebrain in response to transient hypoxia.

To address the influence of oxidative stress and defense capacities in the effects of transient hypoxia in the immature brain, the time course of reactive oxygen species generation was monitored by flow cytometry using dihydrorhodamine 123 and 2',7'-dichlorofluorescein-diacetate in cultured neurons issued from the fetal rat forebrain and subjected to hypoxia/reoxygenation (6 h/96 h). Parallel transcriptional and activity changes of superoxide dismutases, glutathione peroxidase and catalase were analyzed, in line with cell outcome. The study confirmed hypoxia-induced delayed apoptotic death, and depicted increased mitochondrial and cytosolic productions of free radicals (+30%) occurring over the 48-h period after the restoration of oxygen supply, with sequential stimulations of superoxide dismutases. Whereas catalase mRNA levels and activity were augmented by cell reoxygenation, glutathione peroxidase activity was transiently repressed (-24%), along with reduced glutathione reductase activity (-27%) and intracellular glutathione depletion (-19%). Coupled with the neuroprotective effects of the glutathione precursor N-acetyl-cysteine (50 microM), these data suggest that hypoxia/reoxygenation-induced production of reactive oxygen species can overwhelm glutathione-dependent antioxidant capacity, and thus may contribute to the resulting neuronal apoptosis.

Characterization of erythrocyte compounds in asphyxiated newborns.

Perinatal hypoxic-ischemic damage remains a major cause of acute mortality in infants. In our study we have shown that ATP-powered calcium pump was degraded in asphyxiated erythrocyte membranes. Moreover, the activity of Ca2+-ATPase, the enzyme that is solely responsible for maintenance of calcium homeostasis in erythrocytes, was reduced by 50% compared to healthy newborns. We have also detected the enhanced lipid peroxidation in asphyxiated erythrocyte ghosts. To elucidate the potential mechanisms of the calcium pump damage, we have examined the effect of peroxynitrite on Ca2+-ATPase purified from adult human erythrocyte membranes. We have concluded that calcium pump is a direct target for peroxynitrite action in vitro. Our results indicate that erythrocyte membrane compounds could be a primary target for asphyxia-induced damage, and the impairment of the plasma membrane Ca2+-ATPase function could be, in part, mediated by reactive oxygen species.

Postasphyxial reoxygenation reduces the activity of Na+/K+-ATPase in the erythrocytes of newborn piglets.

The aim of our study was to determine whether the impairment of Na+/K+ pump is detectable in erythrocytes during hypoxia and reoxygenation. Acute asphyxia was induced in 10 newborn piglets for 1 h by bilateral pneumothorax. The Na+/K+-ATPase activity, Na+, K+ and ATP content of RBCs were determined in baseline condition (paO2: 60.4 +/- 9.3 mm Hg), at the end of the hypoxic period (1 h) (paO2: 30.2 +/- 10.3 mm Hg), then hourly during the reoxygenation phase (2, 3, 4 h) (paO2: 54.8 +/- 9.0, 56.1 +/- 8.7, 57.2 +/- 9.6 mm Hg). The Na+/K+-ATPase activity was constant during the first 3 h. However, it decreased at 4 h (676 +/- 168 versus baseline 833 +/- 141 U, p < 0.05). The highest ATP content was measured also at this point (4.32 +/- 0.57 versus baseline 3.27 +/- 0.45 mmol/l RBC, p < 0.01). The Na+ content was lower at 1 and 2 h (14.0 +/- 1.8; 13.8 +/- 1.2 versus baseline 15.7 +/- 1.2 mmol/100 g Hb, p < 0.05), but later it became normal. Plasma monovalent cationic levels and intracellular K+ content did not alter during the experiment. Our results indicate that the deterioration of enzyme activity occurs within the same time-frame that previously described morphological alterations in brain tissue develop, so the RBC Na+/K+-ATPase activity might reflect the progress of posthypoxial brain damage.

Decrease of brain protein kinase C, protein kinase A, and cyclin-dependent kinase correlating with pH precedes neuronal death in neonatal asphyxia.

BACKGROUND: Acidosis, energy depletion, overstimulation by excitatory amino acids, and free radical-mediated reactions are the major, current concepts for the explanation of damage and death resulting from asphyxia. Impaired protein phosphorylation by protein kinase C represents another mechanism incriminated in cell death. METHODS: We used a nonsophisticated perinatal asphyxia model to study brain (frontal cortex) pH, ATP, protein kinases PKC, PKA, and cyclin-dependent kinase. We used o-tyrosine, a marker for hydroxyl radical attack, and LPO 586, a spectrophotometric assay, to study lipid peroxidation products. The antioxidant enzymes catalase, superoxide dismutase, and glutathione peroxidase were used in the frontal cortex. In addition, a cell death ELISA and histology to evaluate cell death were performed. RESULTS: Brain pH and protein kinases were decreasing with the length of the asphyctic periods, and energy depletion was shown by a drop of ATP levels, whereas no evidence for the involvement of free radical-mediated mechanisms was obtained. Cell death was shown by the cell death ELISA as early as 10 minutes after the asphyctic period, and histologically, cell death could be revealed but not before day 8 after asphyxia. CONCLUSION: Acidosis and/or impaired protein kinases, but not free radical mechanisms, may play a role in the pathobiochemistry of cell death in neonatal asphyxia of the rat.

Perinatal asphyxia-induced changes in rat brain tyrosine hydroxylase-immunoreactive cell body number: effects of nicotine treatment.

Perinatal asphyxia (15-22 min) was induced to male Sprague-Dawley rat pups during the last day of gestation and the surviving pups were sacrificed at 4 weeks of age. Brain sections were stained for tyrosine hydroxylase immunoreactivity and Cresyl violet. With increasing duration of perinatal asphyxia a reduction in the number of tyrosine hydroxylase immunoreactive (TH-IR) nerve cell bodies was found in the locus ceruleus, probably reflecting an increased death of noradrenaline nerve cell bodies. In contrast, perinatal asphyxia (15-20 min) resulted in an increased number of TH-IR nerve cell bodies in the A9 (zona compacta of the substantia nigra) and the A10 (ventral tegmental area) regions of the mesencephalon, probably reflecting an increased survival of dopamine nerve cell bodies. Perinatal asphyxia for longer than 20 min periods reduced the number of TH-IR cell bodies in the 4 week old rat, even below those found in control animals, indicating that when asphyxia is induced for a period leading to almost 100% mortality, a long-term reduction of the number of mesencephalic dopamine neurons is produced. It has previously been shown that a 4 week postnatal nicotine (0.2 micromol/kg per h) treatment counteracts the asphyxia-induced increase in TH-IR cell body number in the substantia nigra and ventral tegmental area. Such nicotine treatment did not influence the reduction in TH-IR cell bodies in the locus ceruleus following 15-20 min of perinatal asphyxia.

Serum enzyme activities in full-term asphyxiated and healthy newborns: enzyme kinetics during the first 144 hours of life.

Little is known about the kinetics of most serum enzymes during the first hours of life, and even less about the effect on such enzyme activities of perinatal hypoxia-ischaemia. It was the aim of the present study to evaluate the serum kinetics of seven differently located cell enzymes in healthy and asphyxiated newborns during the 1st week of life. The serum activities of cytoplasmic and mitochondrial [aspartate aminotransferase (ASAT), creatine kinase (CK), glutamate dehydrogenase (GLDH), lactate dehydrogenase (LDH), and hydroxybutyrate dehydrogenase (HBDH)] and membrane-bound (gamma-glutamyl-transferase and leucine arylaminidase) enzymes were prospectively measured in full-term asphyxiated (n = 49) and healthy (n = 87) newborns during the first 144 h of life. The blood samples were taken serially at five fixed times: 0 (cord), 12, 24, 72, and 144 h postpartum. The asphyxiated newborns had significantly increased serum activities of ASAT, LDH, and HBDH up to 72 h postpartum, whereas healthy newborns showed higher CK and GLDH activities. Only the activities of ASAT, LDH, and HBDH seemed to depend on the oxygen supply of the fetus or newborn. If other causes of increased serum enzyme activities, e.g. liver diseases, haemolytic disorders, tumours, or inborn errors of metabolism, are excluded, elevated serum activities of ASAT, LDH, and HBDH should draw one's attention to a perinatal hypoxic-ischaemic insult of the newborn.

[Urinary N-acetyl-beta-D-glucosaminidase activity in healthy, polycythemic and hypoxic newborn infants]. / Vizelet N-acetyl-beta-D-glucosaminidase aktivitás egészséges, polycythaemiás és hypoxiás újszülöttekben.

Urinary N-acetyl-beta-D-glucosaminidase (NAG) activity was assayed in 20 polycythemic newborns and prematures, together with 50 prematures suffering from hypoxia on the 1st, 2nd, 4th, 14th, and 28th day after birth. The enzyme was also assayed in 101 healthy newborns which provided normal reference values. NAG activity was factored by the creatinine concentration to given an index. There were significant difference in the NAG indices either between full-term and preterm babies or between appropriate for gestational age (AGA) and small for gestational age (SGA) neonates of the normal group. However, NAG excretion on the first day of life was significantly raised in the case of polycythemic newborns. Following partial plasma exchange, on the 14th day the NAG activity returned to the normal range. NAG activities of premature babies suffering from idiopathic respiratory distress syndrome (IRDS) were significantly elevated on the 1st, 2nd, 4th day but fell sharply to the 14th day. NAG activity fell to normal values by the 28th day. These results suggest that the urinary NAG index is a sensitive indicator of the renal tubular damage during the newborn period.

Asphyctic lesion: proliferation of tyrosine hydroxylase-immunoreactive nerve cell bodies in the rat substantia nigra and functional changes in dopamine neurotransmission.

Asphyxia was induced in male rat pups by performing a delayed cesarean section on pregnant Sprague-Dawley rats. Oxygen saturation and heart rate were recorded during induction of asphyxia. Animals were sacrificed at 3 weeks of age. Brain sections were stained for tyrosine-hydroxylase (TH), dopamine-and-cyclic-AMP-regulated-phosphoprotein-32 (DARPP-32) immunoreactivity (IR) and thionein. Increasing time of asphyxia caused a reduction in the number of nerve cell bodies in the CA1 and CA3 regions of the hippocampus reflecting neuronal death. Furthermore, asphyxia resulted in an increased number of TH-IR nerve cell bodies indicative of a proliferation of dopaminergic neurons in the zona compacta of the substantia nigra. Finally, a significant decrease in rearing was observed in asphyctic animals during the habituation phase, as well as following apomorphine-induced (1 mg/kg s.c.) postsynaptic dopamine receptor stimulation. On the other hand, the apomorphine-induced increase in locomotion was enhanced in asphyctic animals. The implications of these findings for hyperkinesia and attention deficits in disorders resulting from asphyxia are discussed.

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.