MR imaging for the quantitative assessment of brain iron in aceruloplasminemia: A postmortem validation study.

AIMS: Non-invasive measures of brain iron content would be of great benefit in neurodegeneration with brain iron accumulation (NBIA) to serve as a biomarker for disease progression and evaluation of iron chelation therapy. Although magnetic resonance imaging (MRI) provides several quantitative measures of brain iron content, none of these have been validated for patients with a severely increased cerebral iron burden. We aimed to validate R2* as a quantitative measure of brain iron content in aceruloplasminemia, the most severely iron-loaded NBIA phenotype. METHODS: Tissue samples from 50 gray- and white matter regions of a postmortem aceruloplasminemia brain and control subject were scanned at 1.5 T to obtain R2*, and biochemically analyzed with inductively coupled plasma mass spectrometry. For gray matter samples of the aceruloplasminemia brain, sample R2* values were compared with postmortem in situ MRI data that had been obtained from the same subject at 3 T - in situ R2*. Relationships between R2* and tissue iron concentration were determined by linear regression analyses. RESULTS: Median iron concentrations throughout the whole aceruloplasminemia brain were 10 to 15 times higher than in the control subject, and R2* was linearly associated with iron concentration. For gray matter samples of the aceruloplasminemia subject with an iron concentration up to 1000 mg/kg, 91% of variation in R2* could be explained by iron, and in situ R2* at 3 T and sample R2* at 1.5 T were highly correlated. For white matter regions of the aceruloplasminemia brain, 85% of variation in R2* could be explained by iron. CONCLUSIONS: R2* is highly sensitive to variations in iron concentration in the severely iron-loaded brain, and might be used as a non-invasive measure of brain iron content in aceruloplasminemia and potentially other NBIA disorders.

A novel assessment system of toxicity and stability of CuO nanoparticles via copper super sensitive Saccharomyces cerevisiae mutants.

CuO nanoparticles (CuO-NPs) toxicity in organisms is contributed mainly through the copper uptake by both the ionic and nanoparticle form. However, the relative uptake ratio and bioavailability of the two different forms is not well known due to a lack of sensitive and effective assessment systems. We developed a series of both copper resistant and hyper sensitive Saccharomyces cerevisiae mutants to investigate and compare the effects of CuO-NPs and dissolved copper (CuCl2), on the eukaryote with the purpose of quantitating the relative contributions of nanoparticles and dissolved species for Cu uptake. We observed the toxicity of 10 mM CuO-NPs for copper sensitive strains is equal to that of 0.5 mM CuCl2 and the main toxic effect is most likely generated from oxidative stress through reactive oxygen species (ROS) production. About 95% CuO-NPs exist in nanoparticle form under neutral environmental conditions. Assessing the cellular metal content of wild type and copper transporter 1(CTR1) knock out cells showed that endocytosis is the major absorption style for CuO-NPs. This study also found a similar toxicity of Ag for both 10 mM Ag-NPs and 0.2 mM AgNO3 in the copper super sensitive strains. Our study revealed the absorption mechanism of soluble metal based nanomaterials CuO-NPs and Ag-NPs as well as provided a sensitive and delicate system to precisely evaluate the toxicity and stability of nanoparticles.

An In Silico Approach for Assessment of the Membrane Transporter Activities of Phenols: A Case Study Based on Computational Models of Transport Activity for the Transporter Bilitranslocase.

Phenols are the most abundant naturally accessible antioxidants present in a human normal diet. Since numerous beneficial applications of phenols as preventive agents in various diseases were revealed, the evaluation of phenols bioavailability is of high interest of researchers, consumers and drug manufacturers. The hydrophilic nature of phenols makes a cell membrane penetration difficult, which imply an alternative way of uptake via membrane transporters. However, the structural and functional data of membrane transporters are limited, thus the in silico modelling is really challenging and urgent tool in elucidation of transporter ligands. Focus of this research was a particular transporter bilitranslocase (BTL). BTL has a broad tissue expression (vascular endothelium, absorptive and excretory epithelia) and can transport wide variety of poly-aromatic compounds. With available BTL data (pKi [mmol/L] for 120 organic compounds) a robust and reliable QSAR models for BTL transport activity were developed and extrapolated on 300 phenolic compounds. For all compounds the transporter profiles were assessed and results show that dietary phenols and some drug candidates are likely to interact with BTL. Moreover, synopsis of predictions from BTL models and hits/predictions of 20 transporters from Metrabase and Chembench platforms were revealed. With such joint transporter analyses a new insights for elucidation of BTL functional role were acquired. Regarding limitation of models for virtual profiling of transporter interactions the computational approach reported in this study could be applied for further development of reliable in silico models for any transporter, if in vitro experimental data are available.

Chronic copper exposure elicit neurotoxic responses in rat brain: Assessment of 8-hydroxy-2-deoxyguanosine activity, oxidative stress and neurobehavioral parameters.

Copper (Cu), one of the essential transition metal acts as a prosthetic group for variety of proteins and metalloenzymes. However, it may be hazardous when administered in excess. Copper induced memory impairment and progression of neurodegenerative diseases have not yet been fully elucidated. The aim of the present study was to investigate the effect of exposure to copper sulphate (10mg/kg and 20mg/kg body weight, daily for 16 weeks) on brain copper concentration, few biochemical parameters indicative of oxidative stress and on different neurobehavioral functions in male Sprague Dawley rats. Copper-administered animals showed significant increase in brain copper concentration and a depleted Ceruloplasmin level. Different neurobehavioral studies revealed impaired memory and motor coordination in copper exposed rat. Spontaneous locomotors activity and depression symptoms were also noted in copper intoxicated rats. 8-hydroxy-2' -deoxyguanosine (8-OHdG) level, one of the predominant forms of free radical-induced oxidative lesions, and has been widely used as a biomarker for oxidative stress, increased in copper treated group. Copper induced oxidative stress in the brain was also evident from the increased lipid per oxidation (LPO) and nitrite level, depletion of reduced glutathione (GSH), and reduced activities of the antioxidant enzymes such as superoxide dismutase (SOD), and catalase. The present study thus suggests a significant correlation between copper induced oxidative stress and changes in neurobehavioral function in rats. The changes were more pronounced in animals exposed to a higher dose of copper (20mg/kg) than the lower dose.

Copper economy in Chlamydomonas: prioritized allocation and reallocation of copper to respiration vs. photosynthesis.

Inorganic elements, although required only in trace amounts, permit life and primary productivity because of their functions in catalysis. Every organism has a minimal requirement of each metal based on the intracellular abundance of proteins that use inorganic cofactors, but elemental sparing mechanisms can reduce this quota. A well-studied copper-sparing mechanism that operates in microalgae faced with copper deficiency is the replacement of the abundant copper protein plastocyanin with a heme-containing substitute, cytochrome (Cyt) c6. This switch, which is dependent on a copper-sensing transcription factor, copper response regulator 1 (CRR1), dramatically reduces the copper quota. We show here that in a situation of marginal copper availability, copper is preferentially allocated from plastocyanin, whose function is dispensable, to other more critical copper-dependent enzymes like Cyt oxidase and a ferroxidase. In the absence of an extracellular source, copper allocation to Cyt oxidase includes CRR1-dependent proteolysis of plastocyanin and quantitative recycling of the copper cofactor from plastocyanin to Cyt oxidase. Transcriptome profiling identifies a gene encoding a Zn-metalloprotease, as a candidate effecting copper recycling. One reason for the retention of genes encoding both plastocyanin and Cyt c6 in algal and cyanobacterial genomes might be because plastocyanin provides a competitive advantage in copper-depleted environments as a ready source of copper.

Assessment of oxidative stress in serum by d-ROMs test.

Assessment of oxidative stress is an important but technically challenging procedure in medical and biological research. The reactive oxygen metabolites (d-ROMs) test is a simple assay marketed for analyzing the total amount of hydroperoxides in serum via the Fenton's reaction. Earlier reports have raised a suspicion that a part of the signal detected in the assay comes from sources other than metabolites generated by oxidative stress. The aim of this study was to identify which serum components interfere with the d-ROMs signal. By application of sodium azide, ethylenediaminetetraacetic acid, sodium dodecylsulphate, varying temperature, and spiking endogenous substances we demonstrate that in the case of mammalian sera the assay determines ceruloplasmin (CP) activity with potential interferences from hydroperoxides, iron level, thiols, and albumin. In sera of avian species hydroperoxides contribute more to the test outcome, but the CP part is insensitive to inhibition by azide. In conclusion, this assay has deficiencies in terms of detecting realistic concentrations of hydroperoxides, is mostly measuring CP and is also interfered with other serum components, making it very difficult to interpret in most biological systems.

[Endogenous intoxication and biochemical protection in children with celiac disease: state assesment and correlation-regression analysis].

AIM: The purpose of the study was to determine the characteristics of endogenous intoxication parameters, biochemical protection and reveal their interaction in children with celiac disease. MATERIALS AND METHODS: 81 children aged from 1 to 16 years with celiac disease were examined in acute and remission periods. In erythrocytes, blood serum and urine we determined low and moderate molecular weight substances (LMMWS), oligopeptides OP); in erythrocytes--the value of erythrocyte mechanical hemolysis (MH), malondialdehyde (MDA) content, the activity of glutathione reductase GR) and superoxide dismutase (SOD); in blood serum--ceruloplasmin (CP) level, alcohol dehydrogenase (ADH) activity; in erythrocytes and blood serum--glutathione transferase (GT), and calculated intoxication index (II). RESULTS: In children with celiac disease in acute and remission periods LMMWS, OP, II levels in blood were statistically significantly high, while LMMWS level in urine was low. In both periods MH activity was high (p < 0.001), and GSR (p < 0.001) and SOD (p < 0.01) levels were low. We revealed the correlation between MDA and II (r = 0.67; p = 0.006), erythrocyte LMMWS and SOD (r = -0.61; p = 0.015), erythrocyte LMMWS and ADH (r = 0.62; p = 0.006), between GT and OP in urine (r = -0.31; p = 0.026), GTand MDA (r = 0.68; p = 0.000), GT and MH (r = -0.46; p = 0.004), between MDA and CP (r = 0.57; p = 0.002) that made it possible to develop the models of dependence of the parameters in relation to each other. CONCLUSION: In celiac disease there is endogenous intoxication. The changes of the first and the second phases of biotransformation, antioxidant protection is an essential factor of the disease pathogenesis, since they have an effect on endogenous intoxication formation that should be taken into consideration in therapy.

Clinical profile, prognostic indicators and outcome of Wilson's disease in children: a hospital based study.

BACKGROUND: Wilson's disease is a common metabolic disease of the tropics, which is treatable, if diagnosed early. In the paediatric group, the manifestations are mainly hepatic. AIMS: The objective was to study the varied presentations of the disease and to evaluate the diagnostic values of conventional tests in children. The prognostic importance of different indices in liver disorders was also assessed. METHOD: The prospective work was carried out in the Paediatric Medicine Department of Nilratan Sircar Medical College & Hospital (NRSMCH) over a span of three years on children 1 through 12 years of age who fulfilled the prerequisite inclusion criteria. RESULTS: The mean age of the 34 children was 7.7 +/- 2.13 years. Predominant liver involvement was seen in 17 patients, neurological disturbance in 7 and purely hematological manifestations in 2 cases; the remaining 8 children were incidentally diagnosed whilst screening the siblings of affected subjects. In our series the sensitivity of various diagnostic tests was: 24 hour urinary copper excretion--100%, serum ceruloplasmin less than 20 mg/ dL--82.3%, K-F rings--32.35%. Eighteen of the 23 followed up cases (78.2%) responded to medical treatment. The sensitivity and specificity of the new Wilson Index was more than the Nazer index in predicting mortality with liver involvement. CONCLUSION: The superiority of the new Wilson Index over the Nazer index has to be validated on a larger scale. As the outcome of management was very promising, a high index of suspicion in pertinent cases can not only check mortality, but also prevent florid manifestations.

The assessment of blood copper status in cattle: a comparison of measurements of caeruloplasmin and elemental copper in serum and plasma.

AIM: To evaluate the effect of test, either copper (Cu) concentration or caeruloplasmin (CP) activity, and sample type, either serum or plasma, on the diagnosis of blood Cu status in cattle. METHODS: Paired serum and heparinised plasma samples taken from 125 cattle in 13 herds were tested for Cu concentration and CP activity. The individual results for serum Cu concentration and serum and plasma CP activities were compared with the plasma Cu concentration results, as were their diagnostic values as determined by reference ranges, i.e. 'marginal', 'adequate', 'excess'. RESULTS: The overall mean serum Cu concentration was 2.92 micromol/L lower than the mean plasma Cu concentration; however, there was significant variability between individual samples, and the 95% limits of agreement ranged from 0.44 micromol/L more to 6.28 micromol/L less. The relationship between CP activity and plasma Cu concentration was less variable; the 95% prediction interval for plasma Cu concentration from CP activity was +/- 2.8 micromol/L, and was unaffected by whether CP activity was measured in plasma or serum. Using the threshold currently recommended for 'marginal' status of <8.0 micromol/L for serum Cu concentration identified a significantly different population of cattle than a threshold of <9.0 micromol/L for plasma samples. Altering the threshold to <7.0 micromol/L for serum Cu concentration produced better agreement. For CP activity, a threshold of 15 IU/L for both serum and plasma identified the same population as a threshold of <9 micromol/L for plasma Cu concentration. CONCLUSIONS: Serum Cu concentration is not a suitable substitute for plasma Cu concentration for the detection of 'marginal' blood Cu status in cattle as the individual variability in the apparent loss of Cu during clotting is too great. In this study, CP activity, in both serum and plasma, was found to be a suitable substitute for the detection of 'marginal' blood Cu status. CLINICAL RELEVANCE: The use of serum Cu concentration rather than plasma Cu concentration in the diagnosis of Cu responsive disease in cattle needs to be re-evaluated as does the way in which individual sample results are used in such tests.

A kinetic method amenable to automation for ceruloplasmin estimation with inexpensive and stable reagents.

OBJECTIVES: To develop a kinetic, inexpensive, automatable assay for ceruloplasmin as ferroxidase. DESIGN AND METHODS: Dithiothreitol has been used to stabilize the substrate and quinolones to complex the ferric. RESULTS: Mean ferroxidase activity in healthy adults, Wilson's disease and pregnancy was 787.29, 178.5 and 1828.09 IU/L, respectively. Correlation coefficient of ferroxidase vis-à-vis copper, ferroxidase by ferrozine and ceruloplasmin were 0.90, 0.94 and 0.92, respectively. CONCLUSIONS: Norfloxacin method is a simple, inexpensive and automatable kinetic assay.

Functional assessment of the carboxy-terminus of the Wilson disease copper-transporting ATPase, ATP7B.

The carboxy-terminus of ATP7B, the protein defective in the copper-transport disorder Wilson disease, was investigated with respect to its role in copper delivery to the ferroxidase ceruloplasmin. We use yeast as a model system to assess the functional capabilities of ATP7B variants. The yeast ferroxidase, Fet3p, acquires copper from Ccc2p and cannot function if Ccc2p is impaired; expression of wild-type ATP7B in ccc2 yeast complements the iron-deficient phenotype. Our results demonstrate that the C-terminus of ATP7B is necessary for protein stability, as removal of the nonmembranous terminus leads to reduced protein levels and cessation of growth in iron-limited medium. Growth is partially restored when an additional three amino acids are present and is near wild-type levels when only one-third of the C-terminus is present. Measurement of ferroxidase activity is a more sensitive indicator of copper transport function and allowed identification of impaired variants not detected with the growth assay.

Assessment of copper status in epileptic patients treated with anticonvulsant drugs by measuring the specific oxidase activity of ceruloplasmin.

Significant increases in serum levels and decreases in hair copper levels have been previously described in epileptic patients treated with anticonvulsant drugs. A condition not directly related to copper nutriture, such as chronic treatment with these drugs, could increase the serum concentrations of copper and ceruloplasmin and would mask a possible copper deficiency produced by drug-increased biliary copper excretion. Serum immunoreactive ceruloplasmin concentration and its oxidase activity were determined in 90 adult epileptic patients treated with phenobarbital (n=60), phenytoin (n=70), carbamazepine (n=33) and valproic acid (n=8). The levels of ceruloplasmin and oxidase activity were significantly higher (P<0.001) than in an age and gender-matched control group (n=49). The significant correlations (P<0.01) between ceruloplasmin and the urinary excretion of D-glucaric acid, serum gamma-glutamyltransferase (GGT) and drug score in the patients group, would suggest that phenobarbital-type enzyme-inducing agents may increase the hepatic synthesis of ceruloplasmin. In 11 patients with a beta-globulin migrating GGT isoform (GGT3), a sensitive marker of cholestasis, the levels of ceruloplasmin, oxidase activity and total GGT activity were significantly higher (P<0.05) than in the group of 79 patients without the GGT3 isoform; consequently, in some cases a drug-induced cholestasis may also contribute to the increase of serum copper and ceruloplasmin. The values obtained for the specific oxidase activity of ceruloplasmin (activity per unit mass of enzyme protein) suggest that in the most of the cases, chronic administration of phenobarbital, phenytoin, carbamazepine or valproic acid, does not produce marginal or moderate copper deficiency.

Costs of gestation in an Arctic ruminant: copper reserves in muskoxen.

The transfer of trace minerals between mother and fetus may be critical for survival of young ruminants especially among species at high latitudes, which gestate during a long winter and grow through a brief summer. We examined the distribution of copper and metalloproteins (ceruloplasmin and metallothionein) in muskoxen and their fetuses, three times during gestation. Hepatic levels of copper were high in mothers (179 microg g(-1) whole tissue) and did not change through gestation, whereas fetuses accumulated large reserves of Cu (>300 microg g(-1)), likely stored in proteins such as metallothionein, during the last third of gestation. The effect of fetal Cu demands on the pregnant female was tested by supplementation of Cu by subcutaneous injections of Cu gluconate (30 mg Cu/week) during pregnancy. Maternal copper supplementation did not significantly increase hepatic Cu in newborns (412 microg g(-1) for supplemented vs. 303 microg g(-1) for unsupplemented neonates), probably because the diet was already adequate in copper (14 microg g(-1) dry matter). Ceruloplasmin activity declined in pregnant muskoxen that had not received injections of Cu and suggested increased systemic demands for copper during late gestation. Supplies of Cu to the fetus could be limited either by low levels of Cu in the maternal liver, or in the maternal diet during late winter when fetal gains in mass and liver Cu are greatest.

Assessment of microvascular leakage via sputum induction: the role of substance P and neurokinin A in patients with asthma.

Microvascular leakage is an important feature of inflammation. However, the assessment of vascular leakage has seldom been used to monitor airway inflammation in asthma. The aim of this study was to determine the effect of inhaled substance P, a potent neurokinin 1 (NK1) agonist and mediator of plasma extravasation, on markers of microvascular leakage in induced sputum from patients with asthma. In a crossover study, sputum was induced before and 30 minutes after inhalation of substance P or neurokinin A (as control) by 12 subjects with atopic and mild, steroid-naive asthma. The levels of alpha2-macroglobulin, ceruloplasmin, albumin, and fibrinogen were determined in induced sputum as markers of leakage. Substance P induced a significant increase in the levels of alpha2-macroglobulin, ceruloplasmin, and albumin in induced sputum (median fold change, 3.1, 2.2, and 2.9, respectively) (p < 0.013), whereas inhaled neurokinin A was not able to induce significant changes (p > 0.31). The increase in sputum leakage markers was not associated with the cumulative dose of substance P (p > 0.12). These results indicate that NK1 receptor stimulation causes a rapid increase in microvascular leakage as shown in induced sputum in patients with asthma. This investigational model of "dual induction" (first leakage, then sputum) may therefore be useful to test the antiexudative effect of newly develop drugs, such as NK1 antagonists.

Assessment of copper status in pregnancy by means of determining the specific oxidase activity of ceruloplasmin.

BACKGROUND: Conditions not directly related to copper nutriture, such as pregnancy, infections and inflammation, which increase serum copper concentration even during copper deprivation, may be expected to conceal changes in copper status. It has been suggested that the specific enzymatic activity of ceruloplasmin (activity per unit mass of enzyme protein) may be a sensitive indicator of copper status and is not affected by factors such as hormones or sex. In this study, we investigated the behaviour of specific oxidase activity of ceruloplasmin and the copper/ceruloplasmin ratio in pregnant women. METHODS: Copper, immunoreactive ceruloplasmin and its oxidase activity were determined in serum from 52 women in the last trimester of normal pregnancy, and in 50 control women of similar age living in the same area and who were not taking oral contraceptives. The results are expressed as mean+/-S.E.M. RESULTS: In the group of pregnant women, significantly higher serum levels of copper, ceruloplasmin and its oxidase activity were found than in the control group (p < 0.001). In both groups, a high correlation was found between these biochemical variables (r > or =0.905, p < 0.001). However, in the group of pregnant women the specific oxidase activity for ceruloplasmin (364.4+/-3.3 vs. 407.5+/-3.8 U/g) and the copper/ceruloplasmin ratio (2.82+/-0.03 vs. 3.19+/-0.04 microg/mg) were significantly lower than in the control group (p < 0.001). CONCLUSIONS: Although pregnancy accelerates the rate of ceruloplasmin protein synthesis and release with an increase of serum copper, the decrease in specific oxidase activity of circulating ceruloplasmin would be an indicator of the degree of depletion of the mother's copper deposits in order to deal with the foetus' needs.

[Cholestasis assessment by activity of antioxidant enzymes and composition of plasma lipoproteins in patients with liver diseases]. / Otsenka kholestaza po aktivnosti antioksidantnykh fermentov i sostavu lipoproteidov plazmy krovi bol'nykh s patologiei pecheni.

AIM: Investigation of activity of copper-containing enzymes in plasma superoxide dismutase (SOD) and ceruloplasmin (CP) in comparison with concentrations of lipoproteins (LP) of the main classes in patients with chronic hepatic diseases (CHD). MATERIALS AND METHODS: SOD activity, CP and LP in plasma were measured in 90 patients with CHD. RESULTS: An inverse relationship was found between SOD activity and CP content in CHD. SOD/CP ratio proved informative in cholestasis assessment. An increased ratio beta-LP/alpha-LP was noticed in patients with primary biliary cirrhosis. This value and the disease severity correlated. CONCLUSION: Patients with cholestatic hepatic lesions exhibited inhibition of enzyme utilization of superoxide radicals in plasma in line with enhancement of CP secretion.

Copper intake and assessment of copper status.

The diagnosis of marginal copper deficiency has not been perfected despite an increased understanding of the physiologic roles of copper. The use of nonstandardized procedures and the effects of factors other than copper nutriture have impeded identification of an ideal indicator of copper nutritional status in humans. A review of studies of experimental copper deprivation conducted in adult humans over the past 12 y indicated that between 1.0 and 1.25 mg Cu/d is needed by adults for copper maintenance for periods of up to 6 mo and that < or = 2.6 mg Cu/d for periods of up to 42 d is not sufficient for recovery from copper deprivation. Copper-containing enzymes in blood cells, such as erythrocyte superoxide dismutase and platelet cytochrome-c oxidase, may be better indicators of metabolically active copper and copper stores than plasma concentrations of copper or ceruloplasmin because the enzyme activities are sensitive to changes in copper stores and are not as sensitive to factors not related to copper nutriture.

Copper in infant nutrition: safety of World Health Organization provisional guideline value for copper content of drinking water.

BACKGROUND: Copper is an essential nutrient for humans. Recently, a limit of 31.48 micromol/l (2 mg/l) was proposed by the World Health Organization as the provisional guideline value for copper content of drinking water. The objective of the study was to determine the tolerance of chronic exposure to drinking water with low or high copper content in infants. METHODS: Healthy infants (n = 128) were randomly assigned to receive drinking water with less than 1.57 micromol/l (<0.1 mg/l) (n = 48) or 31.48 micromol/l (2 mg/l) of copper (n = 80) from 3 to 12 months of age. At 6, 9, and 12 months of age, serum concentrations of copper, ceruloplasmin, and superoxide dismutase; erythrocyte metallothionein; bilirubin; transaminases; and gamma-glutamyl transferase were measured. RESULTS: Small differences in biochemical indexes of copper nutrition were observed between the groups, but there was no evidence of adverse or toxic effects. These findings may be explained by an adaptive response to the higher copper intake, limiting copper absorption, and increasing biliary secretion, as well as by an increase in copper storage. It is also possible that the sensitivity of the biochemical indicators employed to detect differences in copper status is limited. CONCLUSION: No acute or chronic adverse consequences of consuming water with copper content of 31.48 micromol/l (2 mg/l) were detected in infants during the first year of life. The results support the safety of the World Health Organization's provisional guideline value for copper in drinking water during infancy.

[Evaluation of fetal condition in pregnancy complicated by hypertension--biochemical assessment of amniotic fluid. II. Enzymes]. / Ocena stanu wewnatrzmacicznego plodu w nadcisnieniu tetniczym krwi--poprzez badania biochemiczne plynu owodniowego. II. Enzymy.

Sixty two samples of amniotic fluid were submitted to biochemical investigation including 31 samples from women with pregnancy complicated by hypertension (studied group with blood pressure -65 +/- 15/95 +/- 5 mm Hg) and 31 samples deriving from healthy pregnant women (control group with mean blood pressure 118 +/- 10/74 +/- +/- 9 mm Hg). The following parameters of amniotic fluid were measured: 1) aminotransferases: alanine AlAT and aspartate AspAT, 2) alkaline phosphatase (APt) and its thermostable isoenzyme (APh), 3) ceruloplasmin (Crlp), 4) alpha-amylase (alpha-Amy). The study showed pregnancy complicated by hypertension is related to fetal salivary gland's immaturity presenting decreased activity of alpha amylase in amniotic fluid. Amniotic fluids deriving from women with pregnancy complicated by hypertension showed normal activities of AlAT, AspAT, APt, APh and Crlp.

Assessment of copper nutritional status.

Despite increased interest in the role of copper deficiency in clinical problems and an increased understanding of the physiological roles of copper, the diagnosis of a marginal deficiency has not been perfected. The use of non-standardized procedures and the effects of factors other than copper nutriture have impeded identification of the "ideal" indicator of copper nutritional status in adult humans. The specific activity of copper enzymes, or of copper-containing enzymes in blood cells, such as erythrocyte superoxide dismutase and platelet or leukocyte cytochrome c oxidase, may be a better indicator of metabolically active copper stores than the serum concentration of copper or ceruloplasmin, because the enzyme activities are sensitive to changes in copper stores and are not as sensitive to factors not related to copper nutriture. A single index, such as serum copper concentration, is inadequate for assessing the total body copper nutriture of an individual and must be supported by corroborating evidence.