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Biomedical subjects

E John

Publications and source records attributed to E John.

At least 19 recordsLinked to original sources

Effect of nimodipine on systemic, renal, and cerebral haemodynamics after a mild hypoxic-ischaemic insult in newborn piglets.

Because many Ca2+ channel blocking agents are known to cause adverse systemic effects, in this study we tested the haemodynamic side effects of nimodipine with and without a mild hypoxic-ischaemic (HI) insult in a newborn piglet model. Fifteen min after completing a brief period of asphyxia we gave i.v. nimodipine as 5 micrograms kg-1 bolus followed by 0.1 microgram kg-1 min-1 continuous infusion for 105 min in six piglets, while the same treatment was repeated in five animals without preceding HI insult. At third group of six served as sham operated controls. In the HI insult group by 105 min of nimodipine infusion the mean BP dropped by 30% and the cardiac output dropped by 23% of respective baseline. In this group, the renal blood flow dropped between 65% and 77% of baseline and regional cerebral blood flow dropped between 28% and 55% of respective baseline by 45 min and 105 min of nimodipine fashion. In the group with no prior HI insult, 105 min of nimodipine infusion caused no significant changes in these variables. Despite nimodipine infusion, HI insult caused a significant increase in the mean brain water content compared to the two control groups: 89 +/- 3.2% compared to 82.7 +/- 0.5% in the nimodipine control group, and 83.7 +/- 1.7% in the sham group (P < 0.0001). We conclude that while nimodipine therapy without prior asphyxial insult does not cause significant adverse haemodynamic effects, its infusion after even a mild HI insult could cause reduction in renal blood flow and moderate reduction in regional cerebral blood flow, BP, and cardiac output, suggesting a differential effect. A 15 min lag between HI insult and nimodipine therapy may be too long to prevent cerebral oedema.

Animals

Dysosteosclerosis.

A 5 1/2 year old boy presented with delayed development and loss of vision. A skeletal survey showed osteosclerosis consistent with dysosteosclerosis. Some unusual features of dysosteosclerosis are discussed.

Bone and Bones

Bacterial colonization of indwelling vascular catheters in newborn infants.

OBJECTIVE: To determine the incidence of bacterial colonization of intravascular catheters, to compare the incidence of colonization of intra-arterial (IA), intravenous (IV) and central venous catheters (CVC), and to determine the association, if any, between catheter withdrawal and bacterial sepsis. METHODOLOGY: A prospective observational study was carried out at the neonatal intensive care unit of a university-affiliated regional referral centre. A total of 155 catheters (45 IA, 54 IV and 56 CVC) were obtained from 96 infants admitted to the unit and the distal 0.75 cm studied under the scanning electron microscope. The adjoining 0.75 cm was cultured for bacteria. RESULTS: Scanning electron microscopy revealed that 46% of catheters had bacteria on the internal surface and 13% had bacteria on the outer surface. Greater numbers of CVC were colonized with bacteria compared to IA and IV catheters (P < 0.01). Bacterial colonization of intravascular catheters was not significantly associated with the duration the catheter remained in situ or local reaction at the site of entry of the catheter. Colonization of the external surface of the catheter was significantly associated with bacterial septicaemia (P = 0.0466). Eighty-three per cent of 155 catheters studied had coagulum on the inner or outer surface. Only 53% of these were colonized with bacteria. Bacterial colonization occurred in the absence of a coagulum in only three instances. Catheter withdrawal was not associated with bacterial sepsis. Lack of coagulum on the internal surface of the catheter was strongly associated with septicaemia during the 7 days after catheter withdrawal. CONCLUSIONS: Although significant numbers of intravascular catheters were colonized with bacteria, only colonization with the external surface was associated with catheter-related sepsis.

Analysis of Variance

Case report: neonatal platyspondylic dwarfism--a new form.

A new type of neonatal bone dysplasia is reported. Unusual radiographic abnormalities (platyspondyly with unique shape of the vertebral bodies, shortening of the long tubular bones without signs of rickets or metaphyseal dysplasia, marked shortening of well-ossified short tubular bones, schneckenbecken-like pelvis with tri-radiate acetabulum, pseudoepiphysis of the right ulna, unusual shape of calcaneus and periarticular calcifications) allow designation of this disorder as a new form of neonatal dwarfism.

Abnormalities, Multiple

Renal actions of endothelin-1 in newborn piglets: dose-effect relation and the effects of receptor antagonist (BQ-123) and cyclooxygenase inhibitor (indomethacin).

Although the effects of endothelin-1 (ET-1) on intact or perfused adult kidney are well understood, its effects in the fetus and the newborn have not been well studied. We examined the effects of infusions of 25, 50, and 100 ng/kg of ET-1 per minute on mean blood pressure (MBP), cardiac index (CI), renal blood flow (RBF), glomerular filtration rate (GFR), and urine volume (UV) in 7- to 10-day-old piglets (n = 24). In addition, the effects of pretreatment with a receptor antagonist (BQ-123) and with a cyclooxygenase inhibitor (indomethacin) were studied in 12 separate piglets. ET-1 produced a dose- and level-dependent decrease in CI (60%), RBF (50% to 75%), GFR (66% to 80%) and MBP 15% to 17%. These changes returned to 75% to 80% of baseline 60 minutes after discontinuation of ET-1. Low-dose infusion (25 ng/kg) did not result in any changes in systemic or renal hemodynamics. Plasma half-life of ET-1 in piglets was 2.1 +/- 0.4 minutes. Pretreatment with the specific ETA receptor antagonist BQ-123 completely blocked the ET-1-induced systemic and renal hemodynamic changes. Indomethacin blocked the ET-1-induced rise in MBP but failed to block any renal changes. In fact, indomethacin accentuated the changes induced by ET-1, especially the changes in RBF, RVR, and GFR. Studies of receptor binding in the renal cortex and medulla showed that, in the cortex, the Ki value for ET-1 was 6.32 +/- 1.57, and for ET-3 it was 20.05 +/- 4.38 (p < 0.05); in the medulla, the Ki values were similar for both ET-1 and ET-3. These results indicate that in piglets the renal vascular bed is highly sensitive to ET-1, and its effects are predominantly mediated through ETA receptors.

Animals

Structural perturbations of monoclonal antibodies following radiolabelling: in vitro evaluation of different techniques.

Antibodies are labelled routinely with direct labelling techniques in which some of the native disulphide groups are reduced to sulphydryls. Using polyacrylamide gel electrophoresis (PAGE) as an analytical tool and human polyclonal IgG (HIgG) labelled with 99Tcm by the ascorbic acid (AA), stannous chloride (SnCl2) and 2-mercaptoethanol (2-ME) techniques, we have quantitatively determined any fragmentation of the protein and the radioactivity associated with fragments. HIgG labelled with 99Tcm by two bifunctional chelating agents provided the vital comparison. The results indicated that in the bifunctional chelating agent (BFCA) methods 65 and 61% of the activity was associated with protein with an apparent molecular weight of 150,000 D. The corresponding numbers for the AA, SnCl2 and 2-ME methods were 46.4, 21 and 3.9%. With 2-ME, 96% of the activity was associated with low molecular weight fragments. Although monoclonal antibody fragmentation may not affect immunoreactivity of the protein, it may influence biodistribution.

Antibodies, Monoclonal

Retinopathy of prematurity in infants less than 29 weeks' gestation at birth.

From January 1986 to December 1991 we examined the eyes of 206 infants born at Westmead Hospital, Neonatal Intensive Care Unit who were less than 29 weeks' gestation at birth to determine the incidence of retinopathy of prematurity. Eighty-five infants (41.3%) had no retinopathy of prematurity (ROP) in either eye, 82 infants had stages 1 or 2 ROP (39.8%), 29 had stage 3 ROP (14.1%) and 11 had stage 4 ROP (5.3%). Of these, cryotherapy was performed in 18; six now have bilateral retinal detachment and are blind. The more severe stages of ROP were significantly associated with an increase in the number of days of oxygen supplementation, an increase in the number of days of mechanical ventilation and the presence of patent ductus arteriosus. Infants receiving steroids for mechanical ventilator dependence had a significantly greater chance of requiring cryotherapy (11 or 22 receiving steroids versus seven of 43 without steroids; P < 0.01).

Blindness

Technetium-99m-labeled monoclonal antibodies: influence of technetium-99m binding sites.

UNLABELLED: A number of 99mTc-labeled monoclonal antibodies (Mabs) are being evaluated for diagnostic applications. Preparations are carried out using both direct and indirect (bifunctional chelating agent, BFCA) labeling procedures in which nonspecific 99mTc binding has been postulated. METHODS: By using the ascorbic acid (AA) reduction mediated direct labeling procedure and diaminotetrathiol (N2S4) as a BFCA method, we examined the role of specific binding sites and aliphatic E amino groups of lysine as nonspecific binding sites for 99mTc. RESULTS: Labeling yields for the direct and N2S4 "regular" preparations averaged 73% +/- 2.8% and 91% +/- 2%, for the "specific" preparations, 60% +/- 3.5% and 75% +/- 2% and for the "nonspecific" preparations 13% +/- 1.0% and 16% +/- 1% respectively. All preparations were evaluated in tumor-bearing mice. The control and specific preparations permitted excellent scintigraphic visualization of tumors; the percentages of specific preparations in the tumors being 2.1% +/- 0.5% and 3.1% +/- 0.4%, respectively. With nonspecific preparations, tumors were not visualized and the percentages of administered radioactivity per gram of tumor were only 0.9% +/- 0.2% and 0.9% +/- 0.3%, respectively. CONCLUSIONS: In these 99mTc labeling procedures, the amino group-mediated nonspecific binding of 99mTc to Mabs can be as high as 16% and contributes to increased liver uptake and decreased tumor uptake of radioactivity.

Animals

Short-term outcome of mechanically ventilated infants weighing more than 2499 g at birth: a population based study.

Very little data exist describing the neonatal outcome of infants of birthweight 2500 g or more who require mechanical ventilation. Our aim was to collect population-based data on such infants in New South Wales (NSW), and to monitor their neonatal morbidity, mortality to 1 year of age and the associated risk factors. The study group (NICUS infants) comprised all 341 infants weighing > 2499 g who were admitted to the seven neonatal intensive care units in New South Wales and mechanically ventilated for 4 h or more between 1 January and 31 December, 1987. Two groups of infants emerged: those who were preterm and mostly had hyaline membrane disease, and term and post-term infants for whom the most common problem was 'perinatal asphyxia'. The most important factors associated with dying were a birthweight of over 3499 g (OR = 2.6; CI 1.03-6.6) and a 1 min Apgar score < 4 (OR = 4.8; CI 1.4-16.9). Study group mothers were significantly more likely than all NSW mothers to have had a spontaneous abortion in the previous pregnancy (P < 0.01), a pre-existing medical condition or an obstetric complication in this pregnancy, or a Caesarean section for this delivery (P < 0.001). This is the first population-based study of high-risk neonates without congenital anomalies to clearly document the worsening prognosis associated with a birthweight over 3499 g. Further research should be directed towards identifying prenatal and perinatal factors which might minimize the morbidity and mortality in this group of babies.

Apgar Score

Surfactant replacement therapy in neonates less than 32 weeks gestation: effect on neonatal intensive care resource utilization.

The effect of synthetic surfactant (Exosurf) replacement on complications from hyaline membrane disease (HMD) in infants < 32 weeks gestation and their resource utilization within a neonatal intensive care unit was studied in 1991-92. A control group was selected from infants admitted to the same unit during the preceding 3 years when Exosurf was not available. The infants were controlled for gestation, weight and severity of HMD. Infants given Exosurf had a significant reduction in the incidence of pulmonary interstitial emphysema (PIE), and a marginal decrease in the incidence of pneumothorax. They required fewer days on the ventilator and consumed less of the scarce financial resources. There was no difference in the mortality rate among the two groups. The changes seen were more evident among those infants between 30 and 31 weeks gestation, compared to those < 28 weeks.

Case-Control Studies

Rhenium-186-labeled monoclonal antibodies for radioimmunotherapy: preparation and evaluation.

Rhenium-186 has been determined to be a leading radionuclide for radioimmunotherapy. However, the use of 186Re has been limited due to the lack of a convenient and efficient method by which the radionuclide can be bound to monoclonal antibodies. We have developed a simple technique to label IgM, IgG, fragmented antibodies and tumor necrosis factor-alpha with 186Re. This technique uses ascorbic acid (AA) for controlled reduction of antibody disulfide groups to sulfhydryls and SnCl2 in citric acid for the reduction of 186ReO4-. The labeling yields as determined by instant thin-layer chromatography, molecular filtration and gel filtration were greater than 95% and the colloid formation was less than 5%. The labeled antibodies were stable when challenged with 100 and 250 molar excess of DTPA and HSA for 24 hr at 37 degrees C. SDS-PAGE analysis and autoradiography of labeled IgM, IgG and F(ab')2 monoclonal antibodies indicated uniform labeling and that no fragmentation of the monoclonal antibodies had taken place during the labeling procedure. Immunospecificity of 186Re-labeled human neutrophil specific IgM, as determined by in vitro antigen excess assay, was comparable to that of indium-111-labeled c-DTPA-IgM and technetium-99m-labeled-IgM. A nuclear histone specific 186Re-TNT-1-F(ab')2 was evaluated in mice bearing experimental tumors. The tumor/muscle ratios at 4 and 24 hr were 5.9 +/- 0.21 and 13.8 +/- 6.7, respectively compared to that of 2.4 +/- 0.3 at 4 hr p.i. with a nonspecific protein. The labeling technique is simple, reliable and has already been adapted to a single-vial kit preparation.

Animals

Structure of pentaamminechlororuthenium(III) bisulfate tetrahydrate.

[RuCl(NH3)5](HSO4)2.4H2O, M(r) = 487.88, triclinic, P1, a = 10.0422 (5), b = 14.1044 (7), c = 6.3273 (6) A, alpha = 100.369 (5), beta = 98.655 (6), gamma = 81.354 (4) degree, V = 864.5 (3) A3, Z = 2, Dm = 1.86 (1), D chi = 1.874 g cm-3, lambda(MO K alpha) = 0.71073 A, mu = 13.3 cm-1, F(000) = 498, T = 296 (1) K, R = 0.025 for 4622 unique reflections with I greater than 3 sigma(I). The structure contains (NH3)5RuCl2+ cations, bisulfate anions and solvate water molecules all linked by an extensive hydrogen-bonding network. Each Ru ion is coordinated in a distorted octahedral fashion by five ammonia molecules and a chloride ion. The Ru--Cl distance [2.3742 (5) A] is typical for RuIII--Cl linkages. The Ru--NH3 distances span a narrow range [2.096 (2)-2.119 (2) A] and provide no evidence for a trans effect.

Molecular Structure

Evaluation of biological response modifiers in the enhancement of tumor uptake of technetium-99m labeled macromolecules. A preliminary report.

Imaging tumors with radioactive monoclonal antibodies remains attractive but continues to be challenging. With the hypothesis that the use of biological response modifiers (BRMs) may augment the tumor uptake, technetium-99m(99mTc)-labeled tumor necrosis factor (TNF) and nuclear histone specific TNT-1-F(ab')2 were evaluated in tumor bearing mice given a single dose of interferon (IFN). Ukrain or pokeweed mitogen as BRMs. As early as 1.5 h post injection (p.i.) of the radioactive macromolecules, the absolute tumor uptake (% administered dose/g) of each agent was enhanced (e.g., TNF, control = 1.8 +/- 0.4, Ukrain = 3.2 +/- 0.5, P = 0.006) and tumor to muscle ratios were elevated (e.g., TNF, control a 4.1 +/- 2.2, interferon 8.3 +/- 2.7, P = 0.01). The absolute tumor uptake remained practically unchanged at 4 h p.i. Generally with BRMs, the blood clearance was rapid and tumor/blood ratios and tumor/muscle ratios were higher than in the control group, increasing to greater than 200% for IFN as a BRM. The early enhancement in tumor uptake of macromolecules, leading to excellent delineation of tumors by scintigraphy is highly encouraging and warrants further studies to explore the full potential of BRMs.

Animals

Recovery of tracheal epithelium following high frequency ventilation at low inspired humidity.

We used scanning electron microscopy (SEM) and light microscopy (LM) to study the recovery of tracheal epithelium in newborn lambs damaged by high frequency flow interrupted ventilation (HFFIV) at low inspired humidity (30%). Newborn lambs were mechanically ventilated for 6 h, allowed to recover and subsequently killed at 2, 7 or 14 days. The recovery of the trachea above and below the tip of the endotracheal tube (ETT) was studied at these time periods and compared to a control non-intubated group and a group killed immediately after 6 h of ventilation. Above and below the ETT, SEM and LM revealed deciliation to be greatest 2 days after ventilation. The damaged tracheal mucosa had converted to non-ciliated epidermoid squamous metaplastic cells. Recovery was not complete by 14 days, although the squamous cells had already differentiated into goblet and ciliated columnar epithelial cells. No difference was seen in the rate of recovery of the tracheal mucosa above or below the tip of the ETT.

Animals

A synthetic analogue of melittin aggregates in large oligomers.

An analogue of melittin synthesized in the group of E. T. Kaiser (DeGrado, W. F., F. J. Keźdy, and E. T. Kaiser. 1981. J. Am. Chem. Soc. 103:679-681) was investigated by Raman spectroscopy and fluorescence anisotropy decay. In water, the analogue is completely alpha-helical and aggregates in large oligomers of about 50 monomers. In vesicle membranes, it undergoes orientational fluctuations similar to melittin. The most significant difference from melittin, therefore, is the formation of straight helixes and their aggregation in large oligomers in water. We interpret this as a consequence of the lacking proline residue in the analogue. We, furthermore, hypothesize that the increased tendency for aggregation causes the increased hemolytic activity of the analogue.

Amino Acid Sequence