PubMed HealthSearch

Biomedical subjects

A M Lee

Publications and source records attributed to A M Lee.

14 recordsLinked to original sources

Control of product batches (before and after registration). The British approach.

In the United Kingdom, some pre-registration testing of immunological products is carried out. However, this is normally not extensive. Usually the tests are laboratory tests undertaken to verify results obtained by the manufacturer or quality controller. For many years, routine post-registration testing of batches of immunological products has been carried out. However, this has ceased and had been replaced by a new system known as the Quality Assurance/Quality Control (QA/QC) scheme. The new scheme is designed to provide the Veterinary Medicines Directorate (VMD) with assurance of the continuing quality of batches of immunological products. The basis of the scheme is a system of inspection of companies' quality control facilities and procedures, every eight months, by staff with appropriate microbiological qualifications and experience. Before an inspection is carried out, the company must provide detailed documentation on the facilities and the quality control procedures used for each product. This provides the inspector with the background information for the inspection and discussions with the company.

Animals

Presentation of the EEC directive (81/852)--quality.

The EC directive 81/852 has been revised to include a number of parts setting out the information to be included in applications for marketing authorizations for veterinary immunological products. Part 6 contains the details of the quality aspects which must be addressed by the applicant. It is divided into sections and provides the applicant with the structure and layout to be used for the presentation of the information in the dossier. The applicant must provide sufficient detail to allow the competent authority to assess the quality of the product, including aspects such as batch-to-batch consistency and the risk of contamination of batches with extraneous agents. Thus the dossier must contain details of the starting materials, the method of manufacture, and the controls and tests on the starting materials and intermediate and finished products. The details of the blending and composition must be included. Limits of acceptance should be provided. Procedures used such as inactivation and purification steps and quality control tests should have been shown to be appropriate and the results of the validation studies must be presented. The information supplied must be accompanied by results obtained from batches manufactured by the method described. A justification of the proposed shelf-life of the product must be made with supporting evidence from real-time stability studies.

Animals

Chitin utilization by marine bacteria. A physiological function for bacterial adhesion to immobilized carbohydrates.

Chitin turnover is essential for recycling carbon and nitrogen in marine ecosystems. A key step in this process is the adhesion of marine bacteria to chitin-containing particulates. Vibrio species were therefore surveyed for their ability to bind to immobilized carbohydrates, and one, Vibrio furnissii, adhered to glycosides of three sugars, N-acetylglucosamine (the preferred ligand), D-mannose, and D-glucose. A single Ca(2+)-requiring lectin is responsible for binding to the three sugars. Cells adhering to the chitin analogue divided at the same rate as cells in liquid culture. The first progeny of adherent cells continued to bind to the beads, but the population gradually shifted to a large fraction of free swimming cells, a process that may be necessary for colonization. Metabolic energy is required for cell adhesion to the glycosides, and transient (or no) adhesion occurred in incomplete growth media. The results were explained by studying met and pro mutants. Both the initiation and maintenance of lectin-mediated adhesion requires continuous protein synthesis; expression of lectin activity is a major priority of these cells and functions under conditions adequate for minimal protein synthesis, but insufficient to support cell growth. The adhesion/deadhesion apparatus is apparently used to continuously monitor the nutrient status of the environment, i.e. as a nutrient sensorium. In incomplete medium, cells deadhere, presumably to migrate to a more favorable environment.

Bacterial Adhesion

Chimeric BCR-abl messenger RNA as a marker for minimal residual disease in patients transplanted for Philadelphia chromosome-positive acute lymphoblastic leukemia.

We correlated polymerase chain reaction (PCR)-detectable BCR-abl fusion transcripts with cytogenetic status in 24 patients with acute lymphocytic leukemia (ALL). Of 10 Philadelphia chromosome negative (Ph-) patients, only one was found to exhibit a BCR-abl fusion transcript. Fourteen patients with Ph+ ALL, including eight in clinical remission, exhibited PCR-detectable BCR-abl rearrangements. A detectable Ph chromosome was present in only five of the eight patients in clinical remission. Of the three cytogenetically negative, BCR-abl-positive patients, two eventually succumbed to post-bone marrow transplantation (BMT) relapse. The third died of early transplant complications. Serial PCR analyses were performed on four Ph+ ALL patients in clinical remission who underwent allogeneic BMT. One patient who was PCR negative on post-BMT days 21 and 75 became PCR-positive on day 116 and died in relapse on day 154. One patient was weakly positive for BCR-abl on day 23, negative on day 56, but died of transplant complications on day 124. Two patients exhibited no post-BMT BCR-abl rearrangements and remain well on days 279 and 371. Our findings suggest that PCR analysis may be useful in the early identification of relapse in patients transplanted for Ph+ ALL.

Adult

Epikeratoplasty with nonlyophilized tissue in children with aphakia.

We studied 75 epikeratoplasty procedures using nonlyophilized tissue performed by eight ophthalmic surgeons in 70 eyes (47 patients) to correct for aphakia in children less than 8 years of age (mean age, 3.4 +/- 2.1 years). Of the 47 patients in the study, 24 were girls and 23 were boys; 23 patients had bilateral surgery. Seven of the epigrafts required removal; two were not replaced, and five underwent successful repeat epikeratoplasty. Overall, the success rate (that is, the percentage of epigrafts that remained optically and functionally clear throughout the course of this study) for the epikeratoplasty procedure was 89% (62 of 70 eyes) for initial surgery and 96% (67 of 70 eyes) for repeat surgery. The average spherical equivalent was +14.4 +/- 3.7 diopters preoperatively and +0.3 +/- 2.9 diopters one year after the operation. One year after the final surgical procedure, 42 of 56 eyes (75%) were within 3 diopters of emmetropia. In the 29 verbal patients, best-corrected visual acuity was 20/100 or better in 25 (86.2%) one year after the operation.

Aphakia

Monoclonal IgMs with anti-Gal(beta 1-3) GalNAc activity in lower motor neuron disease; identification of glycoprotein antigens in neural tissue and cross-reactivity with serum immunoglobulins.

IgM monoclonal antibodies (M-proteins) with anti-Gal(beta 1-3)GalNAc and anti-Gal(beta 1-3)GlcNAc activity that bind to gangliosides GD1b and GM1, from two patients with lower motor neuron disease were tested for binding to neural glycoproteins. The M-proteins bound to several glycoproteins in the central and peripheral nervous system including to some in the non-myelin or axonal fraction only. Peanut agglutinin (PNA) which is specific for Gal(beta 1-3)GalNAc, bound to the same protein bands. Since serological studies revealed that the M-proteins were complexed to IgG, serum immunoglobulins were tested for presence of Gal(beta 1-3)GalNAc epitopes. Both PNA and the M-proteins bound to immunoglobulin heavy and light chains, suggesting that the circulating M-proteins bind to Gal(beta 1-3)GalNAc on other immunoglobulins. These studies indicate that in addition to gangliosides the M-proteins might bind to Gal(beta 1-3)GalNAc bearing glycoproteins in vivo and that carbohydrate epitopes on immunoglobulins might have a role in the development and regulation of autoantibodies which cross-react with neural antigens and may cause neurological disease.

Antibodies, Monoclonal

In ovo growth hormone alters growth and adipose tissue development of chickens.

The effect of in ovo administration of ovine growth hormone (oGH) on growth and adipose tissue development of chickens was investigated. Unlike mammalian species, exogenous growth hormone has not been previously shown to increase growth of aves. In trial 1, fertilized eggs were injected with vehicle (.03 M NaHCO3 in .15 M NaCl, pH 8.3), 0.25, 2.5, 25 or 250 micrograms oGH on day 11 of embryogenesis. In trial 2, fertile eggs were injected with vehicle or 250 micrograms oGH. In contrast to previous studies in which GH was administered to growing birds, oGH injected in ovo in the present study increased body weights, skeletal growth and feed efficiencies of male broilers. Growth rate was not altered in females. Adipose cellularity data from both trials indicated that in ovo oGH also altered adipose tissue development of broilers. Seven-week-old male and female broilers treated with oGH during embryogenesis exhibited larger adipocytes with correspondingly less cell per gram of tissue. Additionally, adipocytes from oGH-treated broilers exhibited decreased sensitivity to glucagon, cholera toxin or theophylline-induced lipolysis responsiveness to dcAMP in ovo. Cholera toxin plus theophylline improved the lipolytic response of oGH-treated birds; thus, in broilers injected with oGH cAMP-mediated lipase activation may be reduced by a mechanism of increased phosphodiesterase activity. The results of this study indicate that growth and tissue development of chickens have been altered by mammalian GH in ovo.

Adipose Tissue

Effects of electrical stimulation and an intracellular calcium chelator on calcium movement in suspensions of isolated myocardial muscle cells.

The procedure of Haworth RA, Hunter DR and Berkoff HA (J Mol Cell Cardiol, 1980; 12:715-23) for the isolation of myocardial muscle cells from rat hearts has been modified by the addition of a step which involves centrifugation of the cells through a Percoll gradient. This increased the proportion of rod-shaped cells from 47 +/- 2.2 to 80 +/- 1.3% (mean +/- SEM, n = 7). In the absence of electrical stimulation but in the presence of 1.3 mmol . litre-1 Ca2+ less than 2% of the cells beat spontaneously. This number was not increased by addition of the Ca2+-selective ionophore A23187. A method in which isolated myocytes suspended in a cyclindrical incubation chamber are stimulated to beat by electrical impulses is described. At 1.3 mmol . litre-1 extracellular Ca2+, electrical stimulation increased by 30% the amount of 45Ca2+ exchanged in the period 0.25 to 3 min following addition of 45Ca2+. For myocytes subjected to electrical stimulation, the amount of 45Ca2+ exchanged increased as the concentration of extracellular Ca2+ increased. At 0.5 mmol . litre-1 extracellular Ca2+ verapamil reduced the amount of 45Ca2+ exchanged by 15% while La3+ reduced the amount of 45Ca2+ exchanged by 80%. Incubation of myocytes with the acetoxymethyl ester of the intracellular Ca2+ chelating agent bis (o-aminophenoxy)-ethane-N,N,N'N'-tetraacetic acid (BAPTA) for 45 min led to an inhibition of contraction.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Evaluation of the Oxi/Ferm tube system for identification of nonfermentative Gram-negative bacilli.

The Oxi/Ferm tube system designed for identification of oxidative-fermentative gram-negative bacteria was evaluated for identification of nonfermentative gram-negative bacilli by comparing it with the conventional system. The nine biochemical reactions used as the initial tests in the Oxi/Ferm tube system showed a close agreement with the same conventional reactions. However, the system was only 41% accurate to species or biotype for complete identification of 239 isolates which were well distributed in 48 species and biotypes and included organisms not listed in the Oxi/Ferm tube identification list. The system correctly identified 56% of the test organisms when the degree of identification was based on the manufacturer's guidance list. However, all isolates of Acinetobacter calcoaceticus, Bordetella bronchiseptica, Pseudomonas aeruginosa, P. diminuta, and group IIf were completely identified by the system.

Bacteriological Techniques

Relationship between birth weight and perceptual motor performance in children.

A perceptual-motor performance test (jumping, skipping, balancing, walking) was administered to 150 5- and 6-yr.-old children categorized according to birth weight. Differences were found among the three birth-weight groups, with the children of low birth weight scoring significantly lower than the groups of medium and high birth weight. A substantial positive correlation of .75 was obtained between birth weight and the perceptual-motor performance (total number of items passed).

Birth Weight

A scheme for identification of nonfermentative gram-negative bacteria.

A scheme was designed to speed the identification of 50 nonfermentative gram-negative bacteria to genus, species, and biotype using a minimal numer of biochemical tests. When the scheme was evaluated in our laboratory, one technologist identified 93.6% of 188 known isolates (46 species). Later another technologist, given 161 of the same isolates (46 species) as unknowns, identified 91.9%. The isolates that were misidentified in both cases were atypical organisms with aberrant biochemical reactions, plus 1 typical organism with a delayed oxidation of glucose.

Bacteriological Techniques