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Biomedical subjects

D E Brooks

Publications and source records attributed to D E Brooks.

At least 73 records · Page 4Linked to original sources
PubMed1990

Tonometric and tonographic studies in the normal pony eye.

Tonometric and tonographic techniques were evaluated in the normal pony and compared to other species. Applanation tonometry was utilised to calculate the ocular ridigity of the normal equine eye. The mean intraocular pressure for the pony eye was established as 23.5 mm Hg (sd +/- 4.5). Comparing the non-anaesthetised and anaesthetised pony eye (xylazine-ketamine and glycerol guaiacolate) no significant change in intraocular pressure occurred (P < 0.49). Mean aqueous outflow facility was 0.88 microliter/min/mm Hg (sd +/- 0.65) which is significantly higher (P < 0.01) than the canine and human species. The ocular rigidity values for the pony eye were higher which indicates that the eye is more rigid than either the canine or human eye.

Anesthesia, Local↗
PubMed1990

Biostatistical evidence for two distinct chronic open angle glaucoma populations.

Twenty-six eyes of 26 patients with low-tension glaucoma and 34 eyes of 34 patients with high-tension glaucoma were studied. Fifty-one measurements were available on each patient, including visual field indices, finger blood flow measurements, as well as haematological, coagulation, and biochemical and rheological variables. Multivariate analysis revealed two statistically distinct groups of patients, with low and high tension glaucoma cases equally distributed in both. The smaller group (15 patients) showed a suggestion of vasospastic finger blood flow measurements, and had a high positive correlation between the mean deviation (MD) index of field severity and the highest intraocular pressure (r = 0.715, p = 0.0008). The second, larger group (45 patients) showed disturbed coagulation and biochemical measurements, suggestive of vascular disease, and had no correlation between the MD index and the highest intraocular pressure.

Aged↗
PubMed1990

Evaluation of the Schirmer tear test in clinically normal rabbits.

Mean (+/- SD) value for results of the Schirmer tear test without use of topical anesthesia on 142 rabbit eyes was 5.30 (+/- 2.96) mm/min; range was 15 mm/min, with low value of 0 and high value of 15 mm/min. Two SD from the mean provided a normal clinical range of 0 to 11.2 mm/min for inclusion of 95% of the population. Significant differences were not evident between eye (right vs left) or gender or among breeds; interaction among eye, gender, and breed also was not observed. Although mean values for 2 breeds were significantly different from values in the other 10 breeds and from each other, they were still within the proposed normal range of 2 SD. The Schirmer tear test without topical anesthesia may be useful in rabbits for evaluation of increased values correlated with ocular irritation, rather than for determination of decreased values associated with keratoconjunctivitis sicca.

Anesthesia↗
PubMed1990

Changes in oscillatory potentials in the canine electroretinogram during dark adaptation.

Oscillatory potentials (OP) and electroretinograms (ERG) were recorded from clinically normal dogs after 5, 10, 15, 20, 30, 40, 50, and 60 minutes of dark adaptation. At the end of the adaptation period, OP were characterized by 5 distinct positive peaks, O1 through O5, with mean latencies of 14.46, 20.24, 27.38, 35.31, and 44.85 ms, respectively, and with mean amplitudes ranging from 7.20 to 34.84 microV. After 60 minutes of dark adaptation, the ERG had a mean a-wave latency of 12.03 ms and a mean b-wave amplitude of 109.29 microV. Peaks O3 and O4, which partially mask the summit of the b-wave, had mean latencies of 28.66 and 36.83 ms, respectively. The mean amplitude of the b-wave measured to the peak of O3 was 240.06 microV and 230.73 microV when measured to peak O4. Changes in the OP during dark adaptation consisted of significant (P less than 0.05) increases in the latencies of O1, O2, and O3, and significant increases in the amplitudes of O1, O3, O4, and O5. Concurrent ERG changes consisted of significant increases in the amplitudes of the a-wave and b-wave measured from O3 and O4, and significant increases in the latencies of peaks O3 and O4 on the b-wave.

Animals↗
PubMed1989

Production and isolation of large quantities of monoclonal antibody using serum-free medium and fast protein liquid chromatography.

A method for the production and purification of monoclonal antibody on a large laboratory scale is described. It involves the growth of monoclonal antibodies in serum-free medium, ultrafiltration through an Amicon XM100A filter and fast protein liquid chromatography on a mono Q column with an ionic strength and pH elution gradient. Up to 30 mg of antibody per litre of cell culture supernatant was purified in one day. The procedure can be carried out at 4 degrees.

Animals↗
PubMed1989

Involvement of the MN blood group antigen in shear-enhanced hemagglutination induced by the Escherichia coli F41 adhesin.

An adhesin from Escherichia coli F41 with an apparent subunit molecular weight of 28,000 daltons was isolated by using (NH4)2SO4 precipitation at pH 10 and Sephacryl S-500 gel filtration. The hemagglutination (HA) properties of the native high-molecular-weight adhesin were studied by using a viscometric assay, which provided a quantitative index of the degree of agglutination present as a function of time at a known rate of shear. Shear was found to enhance the degree of agglutination over a 20-min period. A strong, shear-enhanced HA was observed for all donors with the MM or MN blood type studied, but those with the NN blood type showed very little HA. In the microtiter HA assay, the selectivity of the adhesin for MM over NN erythrocytes was found to be dependent on pH and temperature. At 21 degrees C and pH 7.4, there was little difference in HA between the two blood types, but NN cells were progressively more weakly agglutinated than MM cells as the pH or the temperature was increased. Glycophorin A, which bears the M or N determinant, was isolated from individuals with the MM and NN blood types and was shown to be an effective inhibitor of the reaction, with the MM type being the more effective in both microtiter and viscometric assays. Acidic monosaccharides, particularly sialic acid, were also effective inhibitors of HA, although they were less potent on a molar basis than glycophorin. The adsorption isotherm of 125I-labeled adhesin was measured, and the binding was shown to be strongly inhibited by MM glycophorin and somewhat less strongly by NN glycophorin. Collectively, these data strongly suggest that glycophorin AM is a receptor for the F41 adhesin.

Adhesins, Escherichia coli↗
PubMed1989

Scanning electron microscopy of corrosion casts of the optic nerve microcirculation in dogs.

Scanning electron microscopy of vascular corrosion casts of the optic nerve region in normal and glaucomatous Beagles demonstrated that the blood supply to the laminar optic nerve is derived from short posterior ciliary arteries, cilioretinal arteries, and longitudinal pial vessels. The short posterior ciliary arteries formed a ring of striated pillars around the scleral canal. The central retinal artery was not present in the dog. Differences between the casts in normal and glaucomatous dogs were not detected.

Animals↗
PubMed1989

Ultrastructural changes in laminar optic nerve capillaries of beagles with primary open-angle glaucoma.

Ultrastructural examination of optic nerve capillaries in the canine lamina cribrosa revealed many spherical, membrane-bound, electron-dense inclusions that closely resembled Weibel-Palade bodies, in pericytes and endothelial cells of preglaucomatous, early, moderately, and advanced affected Beagles with hereditary primary open-angle glaucoma. This ultrastructural difference between the laminar capillary endothelial cells of normal and glaucomatous Beagles could represent a functional vascular disorder, because Weibel-Palade bodies are associated with microcirculatory abnormalities.

Animals↗
PubMed1989

Morphologic changes in the lamina cribrosa of beagles with primary open-angle glaucoma.

Optic nerve axoplasmic flow is known to be impaired at the scleral lamina cribrosa in Beagles with hereditary primary open-angle glaucoma and is similar to the condition in human beings. Trypsin and detergent digestion to remove all neural, vascular, and glial cellular tissue revealed a well-developed scleral lamina cribrosa in the normal dog in this study. Beagles with primary open-angle glaucoma had signs of mechanical distortion of the anterior lamina cribrosa prior to the detection of ophthalmoscopic changes in the optic nerve head. This change in the supporting architecture of the optic nerve began early and increased in severity as the disease process progressed.

Animals↗
PubMed1988

Column-based separation of erythrocytes using aqueous polymeric two-phase systems.

Cell separation using aqueous polymeric two-phase systems is well established. For separations of cells having similar partition coefficients a multi-step counter-current distribution procedure has been used. As an alternative strategy, we have developed a column-based technique in which the dextran-rich phase of a dextran-polyethylene glycol (PEG) phase system is immobilized onto derivatized chromatography beads. The PEG-rich phase is used as the eluent. A separation of erythrocytes from different species utilizing an affinity ligand was performed. The following behaviour was observed: (i) no elution occurs below a critical affinity ligand concentration; (ii) cell recovery is proportional to affinity ligand concentration; (iii) elution volume is independent of affinity ligand concentration. The above suggest that no multi-step partitioning occurs. However, the separation achieved is considerably better than in a corresponding single-tube separation. A 1:1 mixture of dog and human erythrocytes was separated to a purity of greater than 98% using the column-based technique. The corresponding optimized single-tube separation gave a maximum purity of 84%.

Animals↗
PubMed1988

Column chromatographic separation of cells using aqueous polymeric two-phase systems.

Cell separation using aqueous polymeric two-phase systems is well established. For separations of cells having similar partition coefficients a multistep countercurrent distribution procedure has to be used. However, its operation is limited by time and apparatus constraints. As an alternative strategy we have developed a chromatographic technique in which the dextran-rich phase of a dextran/polyethylene glycol (PEG) phase system is immobilized onto derivatized agarose beads. The PEG-rich phase is used as the eluent. Inclusion of PEG-fatty acid affinity ligand gradients into the eluent produces separations of mammalian erythrocytes based on the differential interaction between the fatty acid and the erythrocyte membranes. A model separation of dog and human erythrocytes has been carried out.

Cell Separation↗
PubMed1988

Development of a general ligand for immunoaffinity partitioning in two phase aqueous polymer systems.

The effect on the partition of erythrocytes in a two phase aqueous polymer system based on dextran T500 and polyethylene glycol (PEG) 8000 of a combination of immunoaffinity ligands, namely, rabbit immunoglobulin G (IgG) and PEG 1900-modified monoclonal IgG, was examined as a potential cell separation technique. Several hybridoma lines secreting mouse monoclonal IgG specific for the Fc receptor of rabbit IgG were raised. The monoclonal IgG was modified by cyanuric chloride attachment of PEG 1900, causing the modified antibody to partition predominantly into the PEG-rich upper phase of the systems. The PEG-modified monoclonal IgG was used as an affinity ligand in the two phase polymer system to specifically increase the partition of rabbit anti-NN glycophorin IgG. The rabbit IgG was applied together with the PEG-modified monoclonal IgG to increase the partition of human erythrocytes. The same system had no effect on the partition of rabbit erythrocytes. These experiments demonstrate that a monoclonal antibody can be modified and used as a general reagent with which to alter cell partition in two phase aqueous polymer systems in an immunologically specific manner.

Affinity Labels↗
PubMed1988

Exogenous methemoglobin as a cyanide antidote in rats.

The effects of the administration of methemoglobin (MetHb) prepared in vitro were evaluated in Sprague-Dawley rats given increasing doses of potassium cyanide (KCN). Median lethal dose (LD50) studies were conducted by giving intraperitoneal injections of KCN (in 0.3- to 0.5-ml volumes), then 2 min later administering intravenous (iv) doses of 1000, 1500, or 2500 mg/kg of MetHb through the tail vein. Control rats received an equivalent volume of saline. The resulting LD50 values for KCN were 7.4 +/- 1.1, 11.7 +/- 1.1, 13.9 +/- 1.0, and 14.2 +/- 1.0 mg/kg (mean +/- SD) for the control (no MetHb) and 1000-, 1500-, and 2500-mg/kg dose groups, respectively. Additional groups of rats were given 1000, 1500, or 2500 mg/kg MetHb and submitted for necropsy. The gross finding of darkened kidneys was present in both dose groups, but became consistent and more prominent in the 2500-mg/kg dose group. Evidence of pathologic changes was not present in other organs. Single-dose pharmacokinetic studies were conducted using iv doses of 1600 and 2500 mg/kg MetHb. The elimination half-life was similar in both doses (62.6 min), but the volume of distribution (95.3 +/- 7.2 and 126.3 +/- 5.2 ml/kg, mean +/- SE) and clearance (1.1 +/- 0.1 and 1.5 +/- 0.1 ml/min/kg) were significantly different (P less than 0.05) for the 1600- and 2500-mg/kg dose groups, respectively. From these data we conclude that although MetHb is cleared from the vascular system rapidly, it may be an effective and nontoxic antidote for doses of cyanide up to twice that of the control LD50.

Animals↗
PubMed1988

Hormonal regulation of rat androgen-binding protein (ABP) messenger ribonucleic acid and homology of human testosterone-estradiol-binding globulin and ABP complementary deoxyribonucleic acids.

Complementary DNA clones coding for rat androgen-binding protein (rABP) were isolated from a rat testis cDNA library constructed in the bacteriophage lambda gt11. The library was screened immunochemically, using two different antibodies against rABP. The identity of the isolated clones was confirmed by epitope selection and DNA sequence analysis. The mRNA encoding rABP could be detected in the testes of 20- and 46-day-old-rats, but not in the 10-day-old rats by hybridization with 32P-labeled rABP cDNA in a Northern blot of poly(A)+-RNA fractioned by agarose gel electrophoresis. No hybridization signal was seen with poly(A)+-RNA isolated from kidney and liver. The rABP mRNA appeared as a single species with a size of 1.65 kilobase, sufficient to encode a protein of 42,000 daltons. The concentration of rABP mRNA in the testes of 37-day-old hypophysectomized rats increased after treatment with testosterone and FSH, given alone or in combination. Sequence and hybridization analysis of cDNAs for rABP, human testosterone-estradiol-binding globulin, and human ABP demonstrates that the cDNAs for human testosterone-estradiol binding globulin and human ABP have greater sequence similarity with each other than either has with rABP.

Amino Acid Sequence↗
PubMed1987

Partition of proteins in aqueous polymer two-phase systems and the effect of molecular weight of the polymer.

The partition of substances in aqueous polymer two-phase systems is influenced by the molecular weight of the phase-forming polymers. We investigate how the effect of the molecular weight of the polymers depends on the molecular weight of the partitioned protein. We show that the magnitude of change of the partition is very small for proteins of molecular weights around 10,000, but increases almost linearly up to molecular weights of 250,000.

Dextrans↗
PubMed1987

Phase partitioning in space and on earth.

In aqueous solution at low concentrations, the neutral polymers dextran and poly(ethylene glycol) (PEG) rapidly form a two-phase system consisting of a PEG-rich phase floating on top of a dextran-rich phase. Biological particles and macromolecules tend to partition differentially between the phases and the liquid-liquid phase interface in these systems. Bioparticle partitioning has been shown to be related to physiologically important surface properties such as membrane charge or lipid composition. Affinity partitioning into the PEG-rich phase can be accomplished by coupling PEG to a ligand having affinity for specific cells or macromolecules. Subpopulations can be identified or separated using multi-step countercurrent distribution (CCD). Incomplete understanding of the influence of gravity on the efficiency and quality of the impressive separations achievable by partitioning, and appreciation for the versatility of this efficient technique, have led to its study for low-gravity biomaterials processing. On Earth, two-phase systems rapidly demix because of density differences between the phases. In low-gravity, demixing has been shown to occur primarily by coalescence. Polymer surface coatings, developed to control localization of demixed phases in low-g, have been found to control electroosmosis which adversely affects electrophoretic separation processes on Earth and in space. In addition PEG-derivatized antibodies have been synthesized for use in immunoaffinity cell partitioning.

Dextrans↗
PubMed1987

Developmental expression and androgenic regulation of the mRNA for major secretory proteins of the rat epididymis.

The concentration of the mRNAs for two pairs of androgen-dependent secretory proteins of the rat epididymis has been measured by filter hybridization with cDNA during the course of post-natal development and following castration and androgen replacement of adult animals. The rate of synthesis of the secretory proteins, measured in an in vitro incubation system, was also ascertained during post-castration involution of the epididymis and during subsequent androgen replacement. The mRNAs for the proteins were first detected at 20 days of age and their levels increased rapidly to reach near adult levels between 30 and 40 days. In the caput epididymidis the levels of the mRNAs declined rapidly during the first week after castration and became undetectable by 16 weeks. Administration of testosterone propionate to animals castrated for 16 weeks reinduced the levels of mRNAs to maximum values by 1 week, but for one pair of proteins the reinduced maximum was only 30% of precastration values. This result, in conjunction with other information, is suggestive that this pair of proteins is under multihormonal control. In the cauda epididymidis, the kinetics of change in mRNA level during androgen withdrawal and replacement were slower than in the caput. In all instances, changes in the rate of synthesis of the secretory proteins followed by observed changes in their level of mRNA, suggesting an absence of androgenic control of translation in the epididymidis.

Animals↗