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Biomedical subjects

J Robinson

Publications and source records attributed to J Robinson.

At least 19 recordsLinked to original sources

A comparative analysis of two preparations of human chorionic gonadotrophin.

A biophysical analysis of two pharmaceutical preparations of human chorionic gonadotrophin (Endo and Profasi) demonstrated that both contained high hCG specific activity. HCG protein analysis by occlusion HPLC and SDS gel electrophoresis showed similar profiles for both products. Although the purified protein components of both preparations were similar their were differences in the contents of the unpurified preparations. The Endo preparation exhibited greater inter-vial variability and contained an unidentified aromatic substance which interacted with the desalting column.

Chemical Phenomena

Activation of neutrophil reactive-oxidant production by synovial fluid from patients with inflammatory joint disease. Soluble and insoluble immunoglobulin aggregates activate different pathways in primed and unprimed cells.

Cell-free synovial fluid from patients with rheumatoid arthritis stimulated the NADPH oxidase activity in human neutrophils, which reached a peak 15-20 min after addition. Insoluble immunoglobulin aggregates isolated from these fluids activated a similar pattern of oxidase activity. However, when synovial fluid was added to neutrophil suspensions which had been previously exposed to granulocyte-macrophage colony-stimulating factor, the stimulated oxidase activity was biphasic, in that an additional transient activity was observed which reached a peak within 5 min of addition. The additional neutrophil-stimulating activity could not be sedimented by centrifugation at 330,000 g-min, and only activated oxidase activity in neutrophils which had previously been primed. The neutrophil-stimulating activity in this soluble fraction was removed by Protein A affinity chromatography, and activity was recovered in eluates from this column. Thus activity in this soluble fraction from synovial fluid is attributed to the presence of soluble immunoglobulin aggregates. Whereas oxidase activity stimulated by the isoluble immunoglobulin aggregates was inhibited by staurosporine (and hence largely dependent on the activity of protein kinase C), the activity stimulated by the soluble immunoglobulin aggregates was staurosporine-insensitive. The soluble immunoglobulin aggregates were present at significantly higher levels in synovial fluids from patients with rheumatoid arthritis compared with those from other joint arthropathies. Thus rheumatoid synovial fluids possess heterogeneous immunoglobulin aggregates which activate neutrophils via distinct molecular pathways. As neutrophils within rheumatoid joints are primed, the soluble immunoglobulin aggregates are likely to be of importance in disease pathology.

Alkaloids

Light and the neonatal eye.

The lighting environment where the baby born prematurely is placed is different from that experienced in utero. As early exposure to light may affect the immature visual system we have attempted to quantify the neonatal ocular light dose. Lighting surveys performed in 7 neonatal units (NNUs) suggested that mean unit illuminance was 470 lux (range 192-890 lux) and intensive care areas within the NNU were significantly brighter than their corresponding low dependency nurseries. The spectral power distribution of fluorescent lights in NNUs was weighted towards the blue end of the spectrum. Datalogging studies demonstrated that between about 30% and 98% of environmental light was incident on the eyelid, which acts as a predominantly red-pass filter, permitting 21% transmission at 700 nm with less than or equal to 5% transmission at 580 nm. Eye-lid opening frequency was quantified: 45% less than or equal to 26 weeks gestational age and decreasing to 7% at 28 weeks gestational age. The onset of the pupillary reflex to light was between 30 and 34 weeks postmenstrual age, the mean diameter was 3.46 mm before this event and 3.02 mm afterwards. Retinal irradiance values calculated from these data show that it is a function of postmenstrual age. Further studies are required to determine its effect on the immature visual system.

Eye

Progressive vision loss. A rare manifestation of familial cavernous angiomas.

We studied four generations of a family in which the index case had progressive loss of vision secondary to a cavernous angioma of the optic nerve and chiasm. Magnetic resonance imaging of the brain revealed multiple, asymptomatic intracerebral cavernous angiomas. Brain magnetic resonance imaging scans of the family members revealed multiple cavernous angiomas in the brother and paternal grandfather, but none in the father or his siblings. Autopsy reports of the paternal great grandfather noted multiple cavernous angiomas in the brain and abdominal viscera. We believe our patient to be the sixth reported case in which a cavernous angioma involved the optic chiasm and optic nerve. Magnetic resonance imaging is a sensitive and specific method of detecting cavernous angiomas. Cavernous angiomas have an autosomal dominant pattern of inheritance with variable penetrance. Surgical intervention in patients with symptomatic cavernous angiomas depends on the location and size of the lesion and associated surgical risks.

Adolescent

PCR amplification of HIV-1 proteinase sequences directly from lab isolates allows determination of five conserved domains.

HIV-1 replication requires limited proteolysis of gag and gag-pol encoded precursor proteins by a specific viral proteinase (PR). Sequences of 20 different HIV-1 strains were compared in order to determine regions of conservation and variability within the PR gene. Viral strains included: (a) five new ones derived from New Orleans patient isolates, (b) four established ones grown in our laboratory, (c) eight, whose sequences were published in the Los Alamos Data Base (1990), (d) one Ugandan, and (e) two Brazilian isolates. In the first two groups, HIV proviral DNA extracted from infected lymphocytes was grown in tissue culture and directly amplified by PCR using specific primers flanking the PR gene. Amplified DNA was directly sequenced using a modified di-deoxy sequencing procedure. Sequence data showed a 25% variation among the 20 different HIV strains studied at the amino acid level, including 8% nonconservative changes and 17% conservative changes. Moreover, five noncontiguous regions were able to be delineated in which the PR showed no amino acid changes. These areas included amino acids (I) 1-9 (amino terminal sequence); (II) 21-32 (sequence around the active site); (III) 47-56 (top of the flap); (IV) 78-88; and (V) 94-99 (carboxy terminal sequence). Our results are consistent with those obtained from X-ray crystallography studies as well as single site mutational analysis.

Acquired Immunodeficiency Syndrome

Improved outcome of respiratory syncytial virus infection in a high-risk hospitalized population of Canadian children. Pediatric Investigators Collaborative Network on Infections in Canada.

PURPOSE: To determine the outcomes in children at high risk for death or complications from respiratory disease who are hospitalized with respiratory syncytial virus (RSV) infection. DESIGN: Retrospective chart review. SETTING: Twelve pediatric tertiary care centers. PATIENTS: All hospitalized children with an RSV infection diagnosed by a positive antigen detection test result or viral isolation during the study period from 1988 to 1991, encompassing three winter seasons. Charts from patients in the following high-risk groups were reviewed in detail: (1) congenital heart disease, (2) chronic lung disease, (3) immunodeficiency, (4) age less than 6 weeks, (5) gestational age less than 36 weeks, and (6) hypoxia (defined as oxygen saturation less than 90% or arterial oxygen pressure less than 60 mm Hg). MEASUREMENTS: The age of all children, the date of RSV identification, and the use of oxygen supplementation, intensive care, and ventilatory support. In addition, the duration of these treatments and the duration of hospitalization were noted. Left-to-right shunting and pulmonary hypertension before RSV infection were determined in those children with congenital heart disease. The nature of the chronic lung disease was noted. Death within 2 weeks of RSV identification was recorded, and the use of ribavirin, bronchodilators, and corticosteroids was determined. RESULTS: Significant year-to-year variation in the frequency of RSV infection was confirmed, with a peak during the 1989-1990 winter noted by the majority of centers (p = 0.0001). Of the 1584 patients in the study, 260 had underlying cardiac disease, 200 had chronic lung disease, 35 had compromised immune function, 378 had been premature, 373 were less than 6 weeks of age, and 338 had hypoxia. Seventeen patients died within 2 weeks (mortality rate 1%); significantly more patients with underlying cardiac disease (3.4%) or lung disease (3.5%) died. Immunocompromised patients had the longest hospital stay (median 39 days), followed by those patients with underlying cardiac or pulmonary disease (11 days); patients less than 6 weeks of age (5 days) and those with hypoxia (6 days) had the shortest hospital stays. Patients with underlying cardiac and pulmonary disease also required oxygen supplementation for a significantly longer period. CONCLUSION: The year-to-year variation in frequency of RSV infection was confirmed in this study. Morbidity and mortality rates associated with RSV infection in a high-risk population in Canada were significantly lower than previously reported.

Adolescent

Myringoplasty for the anterior perforation: experience with the Kerr flap.

Surgical closure of the anteriorly located tympanic membrane perforation can present a problem. The lack of anterior support for the graft frequently leads to graft failure if an underlay method is used whereas anterior blunting is a complication of onlay techniques in this situation. The experience with the Kerr flap, an underlay graft fashioned to include a tab of fascia which is placed laterally under the annulus and the anterior meatal skin, is presented. This method gave a 97.5 percent closure rate with no cases of anterior marginal blunting and a mean auditory threshold gain of 8.5 dB (95 percent confidence limits 5 to 11.9 dB, P less than 0.01) was achieved at the frequencies tested. The use of the Kerr flap is recommended when repairing the anteriorly placed tympanic membrane perforation.

Adolescent

Retinopathy of prematurity: a prospective study. Review at six months.

A cohort of infants of birthweight < or = 1700 g studied prospectively for retinopathy of prematurity (ROP) has been reviewed at 6 months corrected age and the findings related to the neonatal data. The overall incidence of strabismus was 6.4% (30/468), rising from 3.1% (7/229) without ROP to 29.2% (7/24) with stage 3. Strabismus and fusional ability were significantly related to presence and severity of ROP, and abnormal neonatal cranial ultrasound findings. Binocular visual acuity was measured in 340 infants between 20 and 40 weeks corrected age. Eight were subnormal, all due to neurological problems. For the remainder, despite falling within the normal range, there was a significant trend (p < 0.001) for lower acuities with increasing ROP severity. Cycloplegic refraction on 387 infants revealed, with increasing ROP severity: 1, a significant trend towards myopia; 2, increased magnitude of astigmatism; 3, alteration of the astigmatic axis; 4, increased incidence of anisometropia. For the first three categories there was an insignificant trend between no ROP and stage 2, reaching significance only between stage 2 and 3. The predominant axis of astigmatism in infants without ROP was between 60 degrees and 120 degrees, but with ROP there was a significant trend away from this direction.

Age Factors

Light and the immature visual system.

Preterm neonates are cared for in an environment which is continuously and brightly lit, and is very different to that experienced at any other time of life. The amount of light reaching the neonatal eye is governed by two groups of factors. Physical factors include intensity, spectral characteristics and duration of light exposure. Physiological factors are: Frequency of eyelid opening, transmission of light through the closed eyelid, pupil reactivity and area, transmission by the ocular tissues and retinal surface area. Retinal irradiance declines with increasing postmenstrual age. Mechanisms by which light may affect the neonatal eye and clinical conditions in which light have been implicated are considered.

Dose-Response Relationship, Radiation

Natural history of retinopathy of prematurity: a prospective study.

The natural history of retinopathy of prematurity (ROP) has been studied prospectively in 572 infants < or = 1,700 g birthweight using a protocol designed specifically to investigate the subtle features of this condition. Acute ROP developed in 50.9%. All ROP stages 1 and 2 underwent complete resolution and of the 27 (4.7%) infants with stage 3/4 disease, cicatricial sequelae developed in six. Incidence and severity increased with decreasing birthweight and gestational age. Onset was not confined to the temporal retina but exhibited a predilection to start first in the nasal retina of the most immature neonate. The vertical retinal regions were relatively spared. Retinal arteriolar tortuosity developed around three months postnatally and was related to ROP severity but not its presence. The age at onset and rate of progression of retinopathy were largely determined by the stage of development but were also modified by systemic and local factors. The relevance of these findings to ROP screening is discussed.

Acute Disease

Family structure and changes in living arrangements among elderly nonmarried parents.

We used Longitudinal Study of Aging data to investigate change between 1984 and 1986 in living arrangements of older nonmarried parents, focusing on transitions from living alone and living with children to living alone, with children, with others, and institutionalization. Predictors reflect opportunities for coresidence (number and gender of children), resources, needs, and (indirect measures of) attitudes. We find that those with more children are more likely to change from living alone to living with a child. However, number of children does not affect the odds of moving from living with a child to other arrangements (including institutionalization). Gender of children does not affect tendencies to begin coresidence, although there is slight evidence of more movement out of coresidence for those with sons. Findings are interpreted in terms of the influence of adult children's and parents' needs in determining coresidence.

Aged

The use and limitations of Phaneuf's Nursing Audit.

The use of Phaneuf's Nursing Audit in an action research project that is investigating quality assessment and peer review is discussed. A brief description of the project is given followed by an analysis of the audit tool. This analysis suggests that the tool has more disadvantages than advantages. The paper concludes with a discussion about why the tool may be of use despite the disadvantages identified.

Clinical Nursing Research

Relationship of the human immunodeficiency virus type 1 gp120 third variable loop to a component of the CD4 binding site in the fourth conserved region.

Neutralizing antibodies that recognize the human immunodeficiency virus gp120 exterior envelope glycoprotein and are directed against either the third variable (V3) loop or conserved, discontinuous epitopes overlapping the CD4 binding region have been described. Here we report several observations that suggest a structural relationship between the V3 loop and amino acids in the fourth conserved (C4) gp120 region that constitute part of the CD4 binding site and the conserved neutralization epitopes. Treatment of the gp120 glycoprotein with ionic detergents resulted in a V3 loop-dependent masking of both linear C4 epitopes and discontinuous neutralization epitopes overlapping the CD4 binding site. Increased recognition of the native gp120 glycoprotein by an anti-V3 loop monoclonal antibody, 9284, resulted from from single amino acid changes either in the base of the V3 loop or in the gp120 C4 region. These amino acid changes also resulted in increased exposure of conserved epitopes overlapping the CD4 binding region. The replication-competent subset of these mutants exhibited increased sensitivity to neutralization by antibody 9284 and anti-CD4 binding site antibodies. The implied relationship of the V3 loop, which mediates post-receptor binding steps in virus entry, and components of the CD4 binding region may be important for the interaction of these functional gp120 domains and for the observed cooperativity of neutralizing antibodies directed against these regions.

Animals

Discontinuous, conserved neutralization epitopes overlapping the CD4-binding region of human immunodeficiency virus type 1 gp120 envelope glycoprotein.

Monoclonal antibodies have been isolated from human immunodeficiency virus type 1 (HIV-1)-infected patients that recognize discontinuous epitopes on the gp120 envelope glycoprotein, that block gp120 interaction with the CD4 receptor, and that neutralize a variety of HIV-1 isolates. Using a panel of HIV-1 gp120 mutants, we identified amino acids important for precipitation of the gp120 glycoprotein by three different monoclonal antibodies with these properties. These amino acids are located within seven discontinuous, conserved regions of the gp120 glycoprotein, four of which overlap those regions previously shown to be important for CD4 recognition. The pattern of sensitivity to amino acid change in these seven regions differed for each antibody and also differed from that of the CD4 glycoprotein. These results indicate that the CD4 receptor and this group of broadly neutralizing antibodies recognize distinct but overlapping gp120 determinants.

Antibodies, Monoclonal

Role of cAMP-dependent protein kinase in cAMP-mediated vasodilation.

In this study, the role of adenosine 3',5'-cyclic monophosphate (cAMP)-dependent protein kinase A (PKA) in cAMP-dependent relaxation was assessed in the isolated-perfused rat lung using a PKA inhibitor, Rp-cAMPS, 8-bromo-cAMP (8-BrcAMP), and the diterpene activator of adenylate cyclase (AC), forskolin (FSK). A role for K+ channels was also assessed with the nonselective K+ channel blocker, tetraethylammonium (TEA, 10 mM), and an ATP-sensitive K+ channel inhibitor, glibenclamide (GLI, 100 microM). Both 8-BrcAMP (0.1-1.0 mM) and RSK (0.1-10 microM) dose-dependently attenuated the peak pressor response to alveolar hypoxia (HPR). Rp-cAMPS potentiated the HPR and attenuated 8-BrcAMP-mediated vasodilation but had no effect on FSK-mediated vasodilation. FSK-mediated vasodilation was not mimicked by 1,9-dideoxy-FSK, which is biologically inactive on AC but alters K+ channels identically to FSK, nor was it attenuated by the platelet-activating factor antagonist SRI 63-441 or the cyclooxygenase inhibitor indomethacin. TEA, but not GLI, attenuated FSK-mediated vasodilation. Similarly, TEA attenuated 8-BrcAMP-mediated vasodilation. These results support roles for PKA and indirect gating of a non-ATP-sensitive K+ channel in mediating cAMP-dependent pulmonary vasodilation.

8-Bromo Cyclic Adenosine Monophosphate