The gridded retarding ion drift sensor for the petitSat cubeSat mission.

The Gridded Retarding Ion Drift Sensor (GRIDS) is a small sensor that will fly on the 6 U petitSat CubeSat. It is designed to measure the three-dimensional plasma drift velocity vector in the Earth's ionosphere. The GRIDS also supplies information about the ion temperature, ion density, and the ratio of light to heavy ions present in the ionospheric plasma. It utilizes well-proven techniques that have been successfully validated by similar instruments on larger satellite missions while meeting CubeSat-compatible requirements for low mass, size, and power consumption. GRIDS performs the functions of a Retarding Potential Analyzer (RPA) and an Ion Drift Meter (IDM) by combining the features of both types of instruments in a single package. The sensor alternates RPA and IDM measurements to produce the full set of measurement parameters listed above. On the petitSat mission, GRIDS will help identify and characterize a phenomenon known as plasma blobs (or enhancements).

Escherichia coli isocitrate lyase: properties and comparisons.

The glyoxylate cycle was first discovered during studies on bacteria and fungi with the ability to grow on acetate or ethanol as the sole carbon source. Isocitrate lyase, the first enzyme unique to the glyoxylate cycle, has been studied in numerous prokaryotic and eukaryotic organisms. However, information on this enzyme from Escherichia coli is limited. We have recently reported the purification and in vitro phosphorylation of this enzyme. In the present study we have examined and characterized a variety of inhibitors, the divalent cation requirement and the amino acid composition of E. coli isocitrate lyase and compared these results to those obtained with other organisms.

Phosphorylation of isocitrate lyase in Escherichia coli.

Isocitrate lyase from Escherichia coli becomes phosphorylated in vitro by an endogenous kinase when partially purified extracts are incubated with [gamma-32P]ATP. Treatment of isocitrate lyase with histidine modifying reagents, and alkaline hydrolysis of in vitro phosphorylated enzyme indicated the presence of a phosphohistidine residue. Phosphorylation of isocitrate lyase can also occur in vivo, which indicates a possible regulatory significance of this modification. In addition to phosphorylation, isocitrate lyase is capable of incorporating label from both [alpha-32P]ATP and [14C]ATP suggesting that more than one type of covalent modification occurs on this enzyme. This report reviews the studies which have demonstrated the phosphorylation and modification of isocitrate lyase from Escherichia coli.

A chemiluminescent, microparticle-membrane capture immunoassay for the detection of antibody to hepatitis B core antigen.

A chemiluminescent, microparticle-membrane capture immunoassay (CLIA/MMC) for the detection of antibody to hepatitis B core antigen (anti-HBc) is described. The assay utilizes recombinant hepatitis B core antigen coupled to carboxylated latex microparticles. Human polyclonal IgG anti-HBc labelled with acridinium competes with antibody in the sample for a limited number of binding sites on the solid phase. After a 40 min incubation at 40 degrees C, the reaction mixture is transferred to a glass fiber capture membrane and washed. A chemiluminescent signal is produced by addition of alkaline peroxide and is quantitated on a semi-automated reader as described. The CLIA/MMC assay was compared with standard EIA and RIA procedures (Corzyme and Corab, respectively, Abbott Laboratories, North Chicago, IL). Assay sensitivities were RIA greater than CLIA/MMC greater than EIA. A population of 200 normal blood donors showed nearly identical distributions with the CLIA/MMC and RIA (mean = 11% inhibition, SD = 13% for both), compared with the EIA (mean = 13% inhibition, SD = 15%). With a selected plasma population (n = 307), the CLIA/MMC immunoassay showed an excellent correlation (r = 0.94) with both the EIA and RIA procedures. Association of anti-HBc reactivity near assay cutoffs with antibody to hepatitis B surface antigen suggested relative specificity in the order RIA greater than CLIA/MMC greater than EIA. The CLIA/MMC procedure, which can be readily automated, provides a non-istopic alternative to current EIA testing with performance more nearly equivalent to RIA.

Lead in the placenta, membranes, and umbilical cord in relation to pregnancy outcome in a lead-smelter community.

As part of a cohort study of the effects of chronic exposure to lead on pregnancy outcome and child development, lead concentrations in the umbilical cord and placental tissues (body and membranes) from 9 late fetal deaths, 23 preterm births, and 18 births associated with premature rupture of the amniotic membranes were compared with the lead concentrations in the tissues obtained at 22 normal births. Modest elevations in lead concentration were found in the placental body of late fetal deaths (stillbirths) and preterm births as well as in the cord tissue associated with preterm births and premature rupture of membranes. The geometric mean lead concentration in the membranes from late fetal deaths was 2.73 micrograms/g of dry tissue (95% confidence limits 0.69-10.8), which was 3.5 times higher than the mean found in normal births (0.78 micrograms/g, 95% confidence limits 0.61-1.00). The concentration in the membranes of preterm births was also significantly high, being 1.24 micrograms/G (0.91-1.67). Low correlations of membrane and antenatal blood lead concentrations suggest that other factors in addition to exposure to environmental lead may influence the amount of lead accumulated in the placental membranes.

Redefined duplex ultrasonographic criteria for diagnosis of carotid artery stenosis.

OBJECTIVE: To evaluate duplex ultrasonographic criteria for the determination of 50% or more and 70% or more stenosis of the diameter of the internal carotid artery based on conventional angiography in order to align ultrasonographic diagnostic categories with current clinical management schemes. PATIENTS AND METHODS: Between January 1, 1995, and June 30, 1999, 915 patients underwent both carotid duplex ultrasonography and cerebral angiography within 30 days at Mayo Clinic, Rochester, Minn. Of these patients, 294 were excluded from this study because of occlusion of one or both of the internal carotid arteries or atypical flow characteristics. In the remaining 621 patients (61 % male, 39% female; mean age, 67.7 years [range, 14-88 years]), 1218 vessels were available for correlation. Several Doppler ultrasonographic velocity variables were compared with the angiographic findings by use of receiver operating characteristic curve analysis. The primary end point was verification of optimal ultrasonographic criteria to diagnose 70% or more internal carotid artery stenosis. The secondary end point was establishment of threshold values to detect stenosis of 50% or more. RESULTS: At angiography, 382 patients had internal carotid arteries with 70% or more stenosis. Peak systolic and end diastolic velocities of the internal carotid artery and internal carotid artery:common carotid artery peak systolic velocity ratios were measured. For an internal carotid artery stenosis of 70% or more, a peak systolic velocity of 230 cm/s or more resulted in a sensitivity of 86.4%, a specificity of 90.1%, a positive predictive value of 82.7%, a negative predictive value of 92.3%, and an accuracy of 88.8%. An end diastolic velocity of 70 cm/s or more and an internal carotid artery:common carotid artery ratio of 3.2 or more yielded similar values. For an internal carotid artery stenosis of 50% or more, a peak systolic velocity of 130 cm/s or more resulted in a sensitivity of 92.1 %, a specificity of 89.5%, a positive predictive value of 90.3%, a negative predictive value of 91.3%, and an overall accuracy of 90.8%. An internal carotid artery:common carotid artery ratio of 1.6 or more yielded similar values. CONCLUSION: In our ultrasonography laboratory, a carotid artery stenosis of 70% or more (for which carotid endarterectomy is typically recommended in symptomatic patients) is diagnosed reliably with the following duplex ultrasonographic criteria: a peak systolic velocity of 230 cm/s or more, an end diastolic velocity of 70 cm/s or more, or an internal carotid artery:common carotid artery ratio of 3.2 or more.

Mutation analysis of 28 Gaucher disease patients: the Australasian experience.

Gaucher disease is the most common lysosomal storage disease. It is an autosomal recessive disorder that results from a deficiency of beta-glucocerebrosidase. Three clinical phenotypes have been described: non-neuronopathic, acute neuronopathic, and subacute neuronopathic. Genomic DNA from 28 Australasian patients of diverse ethnic origin with Gaucher disease was screened for 3 common mutations (1226G, 1448C and 84GG) using the amplification refractory mutation system (ARMS), and one uncommon mutation (1504T) by restriction enzyme digestion. Thirty-eight of the 56 independent alleles in these patients were characterized, with 1448C present in 42% and 1226G in 28% of the alleles. The 1226G mutation was associated only with the non-neuronopathic phenotype and 7 of the 15 patients who carried the 1448C mutation developed neuronopathic disease. Three infants who died in the neonatal period following a rapidly progressive neurodegenerative course carried no identifiable mutations. The 84GG mutation was carried by 2 Jewish patients and 1504T was present in one patient. It is now possible to rapidly identify the common Gaucher mutations using ARMS and restriction enzyme digestion, and our findings confirm the heterogeneity of mutations in Gaucher disease. It is also possible to predict in part the phenotypic outcome when screening patients for these mutations. We consider mutation analysis to be of most use in prenatal diagnosis and for carrier detection within affected families.

The Port Pirie cohort study: maternal blood lead and pregnancy outcome.

During a three-year period, 831 pregnant women in and around Port Pirie, South Australia--a lead smelter community with longstanding lead pollution--were enrolled in a cohort study to examine prospectively the relation between body lead burden and pregnancy outcome. Three-quarters of the enrolled women were residents of the Port Pirie municipality; the other women lived in adjacent towns and countryside. At 14-20 weeks' gestation, the Port Pirie resident women had a mean blood lead concentration of 10.6 micrograms/dl, while the mean in the other (non-Port Pirie) women was 7.6 micrograms/dl. Similar differences were observed in maternal blood samples taken at 30-36 weeks, at delivery, and from the umbilical cord. These blood lead measures, in conjunction with information collected on other risk factors, were then examined in relation to pregnancy outcome. Among 749 pregnancies followed to completion, pre-term delivery was statistically significantly associated, in a dose-response manner, with maternal blood lead concentration at delivery. Mothers of late fetal deaths (stillbirths) had blood lead concentrations at 14-20 weeks' gestation similar to those of all the other women but had lower concentrations at delivery than the other women. Outcomes of pregnancy for which no association with blood lead was detected were spontaneous abortion, low birthweight (for births at term), intrauterine growth retardation, premature rupture of the membranes, and congenital anomalies.

Port Pirie Cohort study: childhood blood lead and neuropsychological development at age two years.

The Port Pirie Cohort Study is an ongoing prospective study of the relationship between exposure to environmental lead within a lead smelter community, and neuropsychological development in early childhood. Over 600 children, originally recruited during antenatal life, underwent serial blood lead estimations up to two years of age. Systematic interview information was collected on a range of variables, and formal developmental assessment (Bayley Scales of Infant Development) was carried out at 24 months of age. Blood lead concentrations measured antenatally (maternal), at delivery (maternal and umbilical cord) and postnatally at 6, 15 and 24 months were negatively correlated (p less than 0.05) with mental development at 24 months of age. Geometric mean blood lead concentrations (microgram/dl) were 14.3, 20.8 and 21.2 at 6, 15 and 24 months of age respectively. When multiple covariates, including maternal IQ, were controlled for in multiple regression analysis, a statistically significant (p less than 0.01) inverse association was observed between blood lead concentration (PbB) measured at 6 months of age and mental development at 2 years of age. No such association was evident for psychomotor development. When the quality of the home environment (HOME Score) was added to the multiple regression model, the inverse association between blood lead concentration at 6 months of age and mental development at 2 years persisted, albeit less strongly (p = 0.07). From this analysis, it is estimated that a child with with PbB of 30 micrograms/dl at age 6 months will have a deficit of 3.3 points (approximately 3%) on the Bayley Mental Development Scale relative to a child with PbB of 10 micrograms/dl.

Sociodemographic factors modifying the effect of environmental lead on neuropsychological development in early childhood.

A long-term prospective cohort study was conducted to examine the association between prenatal and postnatal exposure to environmental lead and childhood neuropsychological development. The possible interactive effects of blood lead and some covariates on early development were explored in this study. Our data suggest that gender of the child modifies the effect of lead on the neuropsychological development during early childhood. At the ages of 2 and 4 years, girls appear to be more sensitive than boys to the neuropsychological effects of lead. However, there is no significant modification of the effect of lead by some other covariates, such as parental smoking, socioeconomic status, home environment, birth weight, and the kind of infant feeding. Evidence of interactions between environmental lead exposure and other covariates in the causation of neuropsychological deficits in childhood underscores the desirability of considering both main effects and interactions in this area of research. Such effects, if confirmed, may have implications for public health intervention strategies.

Determinants of blood lead concentrations to age 5 years in a birth cohort study of children living in the lead smelting city of Port Pirie and surrounding areas.

Sources of variation and some principal determinants of blood lead concentration (PbB) were investigated in a cohort of children, followed to age 5 y, who were born near a lead smelter in Port Pirie, South Australia. The child's age and place of residence were the two variables most strongly predictive of PbB. A sharp increase in PbB occurred between 6 and 15 mo of age and was followed by a peak concentration that occurred at approximately 2 y of age, after which PbB steadily and consistently declined. Irrespective of age, the PbBs in children who lived in Port Pirie were significantly higher than levels identified in children who resided outside the city. There was no significant difference in PbB between boys and girls. Elevated PbB at each specific age was associated mainly with increased lead concentrations in the topsoil of the local residential area, employment of the father in the lead industry, parental smoking, and behaviors likely to cause ingestion of dirt. Blood samples taken from children at certain ages and during the warmer months contained more lead than samples obtained during the cooler months. The effects of these determinants on PbB during early childhood were basically consistent in both single and multivariable analyses.

Prevalence of neural tube defects in South Australia, 1966-91: effectiveness and impact of prenatal diagnosis.

OBJECTIVE: To determine trends in total prevalence of neural tube defects in South Australia during 1966-91, the impact of prenatal diagnosis on birth prevalence, and the effectiveness of prenatal screening for neural tube defects in 1986-91. DESIGN: All births and terminations of pregnancy affected by neural tube defects and information on prenatal screening were ascertained from multiple sources including the South Australian perinatal and abortion statistics collections, birth defects register, and state maternal serum alpha fetoprotein screening programme. SETTING: Southern Australia. SUBJECTS: All 1058 births and terminations of pregnancy affected by neural tube defects in 1966-91. MAIN OUTCOME MEASURES: Total prevalence and birth prevalence of individual and all neural tube defects. The proportion of screened cases detected prenatally. RESULTS: Total prevalence of neural tube defects during 1966-91 was 2.01/1000 births with no upward or downward trend. However, birth prevalence fell significantly (by 5.1% a year), with an 84% reduction from 2.29/1000 births in 1966 to 0.35/1000 in 1991 (relative risk = 0.16, 95% confidence interval 0.07 to 0.34). The fall was 96% for anencephaly and 82% for spina bifida. 85% of defects, both open and closed, were detected before 28 weeks' gestation in women screened by serum alpha fetoprotein or mid-trimester ultrasonography, or both, in 1986-91 (99.0% for anencephaly and 75.7% for spina bifida). CONCLUSIONS: While the total prevalence of neural tube defects in South Australia remained stable, prenatal diagnosis and termination of pregnancy resulted in an 84% fall in birth prevalence during 1966-91. Screening detected over four fifths of cases in 1986-91.

PIP: The authors sought to determine trends in total prevalence of neural tube defects in South Australia during the period 1966 through 1991, the impact of prenatal diagnosis on birth prevalence, and the effectiveness of prenatal screening for neural tube defects during the period 1986 through 1991. The authors studied 1058 births and terminations of pregnancy affected by neural tube defects during 1966-1991. Data on births and terminations and information on prenatal screening came from the South Australian perinatal and abortion statistics collections, birth defects registers, and the state maternal serum alpha fetoprotein screening program. Main outcome measures of the study were total prevalence and birth prevalence of individual and all neural tube defects and the proportion of screened cases detected prenatally. Total prevalence of neural tube defects during the period 1066-1991 was 2.01/1000 births with no upward or downward trend. However, birth prevalence fell significantly (by 5.1% a year) with an 84% reduction from 2.29/1000 births in 1966 to 0.35/1000 in 1991 (relative risk = 0.16, 95% confidence interval 0.07-0.34). The fall was 96% for anencephaly and 82% for spina bifida. 85% of defects, both open and closed, were detected before 28 weeks gestation in women screened by serum alpha fetoprotein or midtrimester ultrasonography, or both, in 1986-1991 (99.0% for anencephaly and 75.7% for spina bifida). While the total prevalence of neural tube defects in South Australia remained stable, the prenatal diagnosis and termination of pregnancy resulted in an 84% fail in birth prevalence during 1966-1991. Screening detected over four-fifths of the cases during 1986-1991.

Neonatal screening strategy for cystic fibrosis using immunoreactive trypsinogen and direct gene analysis.

OBJECTIVE: To assess the effectiveness of a two tier neonatal screening strategy for cystic fibrosis, which combines estimation of immunoreactive trypsinogen followed by direct gene analysis in dried blood spot samples collected at age 5 days. DESIGN: Prospective study of two tier screening strategy. The first tier of testing immunoreactive trypsinogen concentration was measured in dried blood spot samples from neonates aged 4-5 days. In the second tier direct gene analysis to detect cystic fibrosis mutations deltaF508 and deltaI506 was performed in those blood spot samples which produced the highest 1% of immunoreactive trypsinogen values. Direct gene analysis was also performed on blood spot samples from infants with suspected or confirmed meconium ileus, regardless of the immunoreactive trypsinogen value. SETTING: The South Australian Neonatal Screening Programme, operating from the department of chemical pathology at Adelaide Children's Hospital. Subjects--All 12,056 neonates born in South Australia between December 1989 and June 1990. No selection criteria were applied. INTERVENTIONS: All infants found to have two recognised cystic fibrosis mutations on direct gene analysis were referred directly for clinical management, and those with one recognised cystic fibrosis mutation were recalled for a sweat test; their families were given genetic counselling. MAIN OUTCOME MEASURES: Direct or exclusion of cystic fibrosis by sweat testing of neonates identified as being at high risk of cystic fibrosis on screening and of those at minimum risk but whose subsequent clinical history raised suspicion about the disease. RESULTS: Of the 12,056 infants screened, 11,907 (98.8%) were reported as "cystic fibrosis not indicated" on the basis of low immunoreactive trypsinogen values. Of the 148 (1.23%) infants with raised immunoreactive trypsinogen values and one (0.008%) with meconium ileus, 132 (1.09%) were reported as cystic fibrosis not indicated, four (0.033%) were identified as having cystic fibrosis, and 13 (0.108%) were recalled for sweat testing after direct gene analysis for the presence of the deltaF508 and deltaI506 cystic fibrosis mutations. No cases of affected infants are known to have been missed to date. CONCLUSION: The strategy of measurement of immunoreactive trypsinogen followed by direct gene analysis is a highly specific neonatal screen for cystic fibrosis, requiring only 2.8 families to be contacted for every case of cystic fibrosis diagnosed.

Neonatal screening for cystic fibrosis using immunoreactive trypsinogen and direct gene analysis: four years' experience.

OBJECTIVE: To assess the performance and impact of a two tier neonatal screening programme for cystic fibrosis based on an initial estimation of immunoreactive trypsinogen followed by direct gene analysis. DESIGN: Four year prospective study of two tier screening strategy. First tier: immunoreactive trypsinogen measured in dried blood spot samples from neonates aged 3-5 days. Second tier: direct gene analysis of cystic fibrosis mutations (delta F508, delta I506, G551D, G542X, and R553X) in samples with immunoreactive trypsinogen concentrations in highest 1% and in all neonates with meconium ileus or family history of cystic fibrosis. SETTING: South Australian Neonatal Screening Programme, Adelaide. SUBJECTS: All 88,752 neonates born in South Australia between December 1989 and December 1993. INTERVENTIONS: Neonates with two identifiable mutations were referred directly for clinical assessment and confirmatory sweat test; infants with only one identifiable mutation were recalled for sweat test at age 3-4 weeks. Parents of neonates identified as carriers of cystic fibrosis mutation were counselled and offered genetic testing. MAIN OUTCOME MEASURES: Identification of all children with cystic fibrosis in the screened population. RESULTS: Of 1004 (1.13%) neonates with immunoreactive trypsinogen > or = 99th centile, 912 (90.8%) had no identifiable mutation. 23 neonates were homozygotes or compound heterozygotes; 69 carried one identifiable mutation, of whom six had positive sweat tests. Median age at clinical assessment for the 29 neonates with cystic fibrosis was 3 weeks; six had meconium ileus and two had affected siblings. 63 neonates were identified as carriers of a cystic fibrosis mutation. Extra laboratory costs for measuring immunoreactive trypsinogen and direct gene analysis were $A1.50 per neonate screened. CONCLUSION: This strategy results in early and accurate diagnosis of cystic fibrosis and performs better than screening strategies based on immunoreactive trypsinogen measurement alone.

Para-influenza pneumonia in DiGeorge syndrome two years after thymic epithelial transplantation.

A patient with DiGeorge syndrome developed pneumonia caused by para-influenza virus type 3 two years after immunological reconstitution with foetal thymic eptihelium. There was a transient reduction of mitogen-induced lymphocyte transformation at the time of the pneumonia. Although she recovered from the pneumonia, brochitis persisted and the virus could still be isolated from her pharyngeal secretions 3 1/2 months later.

Evaluation of a state-wide neonatal screening programme.

A screening programme which was already established to detect phenylketonuria in the newborn period in South Australia was extended to include screening for galactosaemia, homocystinuria, hereditary tyrosinaemia, histidinaemia, maple syrup urine disease and severe alpha 1-antitrypsin deficiency for a trial period. Later, screening for hypothyroidism was introduced. Results suggest that screening for galactosaemia and hypothyroidism are useful additions to the programme. Screening for trrosinaemia and alpha 1-antitrypsin deficiency produced a high number of requests for repeat samples, causing anxiety and no positive benefit to patients. Homocystinuria, an eminently treatable condition, was not detected, nor was maple syrup urine disease, a much less readily treatable condition. Histidinaemia was detected only once. Screening for tyrosinaemia, alpha 1-antitrypsin deficiency, maple syrup urine disease and histidinaemia has been discontinued. Newborn screening in South Australia currently includes tests for phenylketonuria, hypothydroidism, galactosaemia and homocystinuria.

Purification and Characterization of Cytosolic NADP Specific Isocitrate Dehydrogenase from Pisum sativum.

Cytosolic NADP-specific isocitrate dehydrogenase was isolated from leaves of Pisum sativum. The purified enzyme was obtained by ammonium sulfate fractionation, ion exchange, affinity, and gel filtration chromatography. The purification procedure yields greater than 50% of the total enzyme activity originally present in the crude extract. The enzyme has a native molecular weight of 90 kilodaltons and is resolved into two catalytically active bands by isoelectric focusing. Purified NADP-isocitrate dehydrogenase exhibited K(m) values of 23 micromolar for dl-isocitrate and 10 micromolar for NADP, and displayed optimum activity at pH 8.5 with both Mg(2+) and Mn(2+).

Evidence of histidine phosphorylation in isocitrate lyase from Escherichia coli.

Escherichia coli isocitrate lyase (EC 4.1.3.1.) can be phosphorylated in vitro by an ATP-dependent reaction. The enzyme becomes phosphorylated by an endogenous kinase when partially purified sonic extracts are incubated with [gamma-32P]ATP. Treatment of isocitrate lyase with diethyl pyrocarbonate, a histidine-modifying reagent, blocked incorporation of [32P]phosphate from [gamma-32P]ATP. The isoelectric point of the enzyme was altered by treatment with phosphoramidate, a histidine phosphorylating agent, which suggests that isocitrate lyase can be phosphorylated at a histidine residue(s). Immunoprecipitated 32P-labeled isocitrate lyase was subjected to alkaline hydrolysis, mixed with chemically synthesized phosphohistidine standards, and analyzed by anion exchange chromatography. Characterization of the phosphoamino acid was based on the demonstration that the 32P-labeled product from alkali-hydrolyzed isocitrate lyase comigrated with synthetic 1-phosphohistidine. In addition, loss of catalytic activity after treatment with potato acid phosphatase indicates that catalytically active isocitrate lyase is the phosphorylated form of the enzyme.