High myopia and pre-eclampsia: a blinding combination.
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Biomedical subjects
Publications and source records attributed to H Cook.
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High-pressure freezing is applicable to both morphological and immunocytochemical studies. We are investigating the morphogenesis of foot-and-mouth disease virus and African swine fever virus by the use of high-pressure freezing of infected cells. Foot-and-mouth disease virus particles are not detected in sections of conventionally immersion-fixed infected cells, but when the cells are prepared by high-pressure freezing, newly formed virions are readily seen throughout the cell. We report two methods for high-pressure freezing of virally infected cells: first, two sapphire discs frozen 'face to face' with a narrow spacer to prevent cell damage and, second, a fibrous filter substrate that can be easily cut into discs to fit into the freezing planchettes. Cells readily adhere to the fibres in vitro, and the complete disc can be rapidly transferred to the planchettes for freezing. Immunolabelling studies of the microneme proteins of the parasite Eimeria tenella indicate that high-pressure freezing followed by freeze-substitution in acetone with uranyl acetate allows high-sensitivity immunolabelling for these proteins.
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This paper describes an electrostatic ion chromatographic system in which the separation selectivity for inorganic anions, especially for sulfate and phosphate, could be manipulated by altering the molar ratio of the zwitterionic and cationic surfactants in the column coating solution used to prepare the stationary phase. The zwitterionic surfactant used for this study was 3-(N,N-dimethyltetradecylammonio)propanesulfonate (Zwittergent-3-14) and the cationic surfactant was tetradecyltrimethylammonium (TTA). Using a reversed-phase C18 column (250x4.6 mm I.D.) coated with 10/10 (mM/mM) of TTA/Zwittergent-3-14 mixed micelles as the stationary phase and either NaHCO3 or Na2CO3 aqueous solution as the eluent, together with suppressed conductivity detection, baseline separation of seven model inorganic anions was obtained. The elution order for those anions was found to be F+ < HPO4(2-) < Cl- < SO4(2-) < NO2- < Br- < NO3-. Under the same conditions but using 1/10 (mM/mM) of TTA/Zwittergent-3-14 mixed micelles as the column coating solution, the elution order for these model ions was F- < HPO4(2-) < SO4(2-) < Cl- < NO2- < Br- < NO3-. The early elution of phosphate and sulfate is a unique attribute of this system. Detection limits for F-, HPO4(2-), Cl-, SO4(2-), NO2-, Br- and NO3- (S/N=3, sample injection volume 100 microl) were 0.11, 0.12, 0.12, 0.18, 0.49, 0.49, 0.52 microM, respectively.
Different species and different strains of animals commonly show very different sensitivities to carcinogenic regimes, which are often unexplained. A major possible contributory factor is variation in susceptibility to mutation, but this has not been directly demonstrated. This study therefore quantified the colonic stem cell mutation frequency in three strains of mice using two carcinogens. Stem cell mutations were identified using loss of function of glucose 6-phosphate dehydrogenase (G6PD) in individual crypts, a technique validated by several previous studies. The carcinogens dimethylhydrazine (DMH) and ethyl nitrosurea (ENU) were given to Balb/C, C57BL/6J, and C3H mice. In response to DMH, Balb/C mice were most susceptible, with approximately double the stem cell mutation frequency found in C3H and more than ten-fold that found in C57BL/6J (3.3+/-0.71 vs. 1.5+/-0.52 vs. 0.28+/-0.8x10(-4)). In response to ENU, Balb/C mice and C3H mice were equally susceptible, showing a stem cell mutation frequency approximately twice that of C57BL/6J (3.1+/-0.4 vs. 3.1+/-0.65 vs. 1.63+/-0.28x10(-4)). The observed differences among the strains with respect to somatic mutation following DMH treatment are likely to be due to the previously documented differences in metabolic conversion to the active metabolite. However, as ENU is a directly acting, rapidly inactivated mutagen, strain differences in response to ENU are unlikely to be due to strain-dependent metabolism of the mutagen and are likely to reflect differences in DNA repair efficiency, or possibly in stem cell kinetics among the strains studied. Susceptibility to the induction of colonic stem cell mutation is an important factor in susceptibility to carcinogens, whether due to differences in DNA repair or to other factors. Direct quantification of stem cell mutation frequency allows the separate identification of this component of the carcinogenic cascade and shows that it can make a major contribution to the differing susceptibility of different mouse strains.
Detection of rare events within solid tissues by immunocytochemistry is aided by imaging thick sections. Sections of 40--100 microm thickness of paraformaldehyde-fixed solid tissue can be prepared by use of a vibrating microtome and when immunolabelled these sections can be imaged in a confocal microscope. This approach provides excellent preservation of the structure of the sample and imposes minimal antigenic damage. In studies of the invasion of the bovine intestinal epithelium by Salmonella, this method has allowed detection of individual invading bacteria within large samples. The thick vibrating microtome sections were also used for the detection of rare apoptotic cell nuclei identified by TUNEL staining.
Chronic leg wounds are characterized by defective remodeling of the extracellular matrix, failure of reepithelialization, and prolonged inflammation. The hypothesis that this defective extracellular matrix remodeling is associated with phenotypic differences in the activity of the matrix metalloproteinases and tissue inhibitors of metalloproteinases was studied in chronic wound and patient-matched normal fibroblasts in three-dimensional collagen lattice systems. Chronic wound fibroblasts exhibited no differences in morphology or proliferation (p > 0.1) compared with patient-matched uninvolved dermal fibroblasts. The ability of chronic wound fibroblasts to reorganize extracellular matrix was significantly impaired, however, in comparison to the uninvolved dermal fibroblasts (p < 0.01). This difference in extracellular matrix reorganization was not related to differences in proliferation within the collagen lattices (p > 0.05) or attachment to type I collagen (p > 0.1). Marked differences were evident in matrix metalloproteinase-2 activity between chronic wound and patient-matched normal fibroblasts. Whereas levels of pro-matrix metalloproteinase-2 were similar between the two fibroblast populations (p > 0.1), the chronic wound fibroblasts exhibited significantly decreased levels of the 62 kDa active form of matrix metalloproteinase-2 (p < 0.01). Reverse zymography and enzyme-linked immunosorbent assay demonstrated that the decreased matrix metalloproteinase-2 activity was associated with increased production of tissue inhibitors of metalloproteinase-1 and -2 by the chronic wound fibroblasts (p < 0.05). Increased production of tissue inhibitors of metalloproteinases in chronic wound fibroblasts was also reflected in decreased levels of matrix metalloproteinase-1 (p < 0.005). These data suggest that the impaired ability of chronic wound fibroblasts to reorganize extracellular matrix in vitro is related to decreased levels of active matrix metalloproteinase-2 and matrix metalloproteinase-1 resulting from increased production of tissue inhibitors of metalloproteinase-1 and -2 by chronic wound fibroblasts. These findings provide a mechanism to explain the impaired cellular responses and extracellular matrix reorganization observed in chronic leg wounds in vivo.
Weanling pigs were used in a series of studies to determine the feeding value of an enzymatically digested protein product developed from a blend of swine and poultry abattoir by-products. The initial study used 156 pigs weaned at approximately 22 d of age to compare the product with menhaden fish meal in Phase II diets. The product supported equal growth rate, and there was no preference for diet exhibited based on inclusion level of the enzymatically digested protein product. The second study used 100 pigs weaned at approximately 21 d of age to compare the product with spray-dried animal blood cells in Phase II diets. The product supported a growth rate equal to that with the blood cells, and the combination of products enhanced growth rate (P<.05). The third study used 265 pigs to compare the product with spray-dried porcine plasma in a slope ratio growth assay. Results demonstrated a relative feeding value of 91% for the product over a 4-wk feeding period. The fourth study used 290 pigs to compare the product with spray-dried porcine plasma in Phase II diets; results demonstrated comparable growth performance. The final study used 180 pigs to compare the product with spray-dried porcine plasma in Phase I diets; results demonstrated comparable growth performance. These data indicate that the enzymatically digested abattoir by-product is a high-quality protein source for weanling pigs.
The present study compared the relationship between motivation for intimacy and level of premorbid adjustment for men and women with schizophrenia. A sample of 34 schizophrenic patients (14 male and 20 female) were studied. Stories told in response to six TAT cards were scored for Intimacy Motivation (IM). Levels of Premorbid adjustment (PA) and Asociality (ASOC) were assessed. A statistically significant interaction between gender and level of PA indicated that males with good PA had higher IM than those with poorer whereas females with good PA had lower IM than females with poor PA. The same interaction was obtained for the measure of ASOC. Findings are discussed in relation to sociocultural expectations for women. Some implications for differential treatment needs of male and female schizophrenics are suggested.
Using the techniques of relaxed stringency polymerase chain reaction and genomic library screening, we have isolated homologous canine and human genes that encode a novel putative seven transmembrane G-protein-linked receptor. The gene encodes an open reading frame (ORF) of 993 bp. The sequences of the canine and human ORFs are highly conserved, sharing 89% nucleotide identity and 92% amino acid similarity between the two species. Northern blot analysis demonstrates that mRNA transcripts of the gene are abundantly expressed in testis and spleen with a lesser degree of expression observed in several other tissues associated with endocrine and immunologic/hematologic function. The gene, designated GPR18, was localized to human chromosome 13q32 using fluorescence in situ hybridization.
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A sexually dimorphic distribution of galanin (GAL)-like immunoreactive (ir) neurons and fibers was found in the brain and pituitary of goldfish. The rostralmost GAL-ir perikarya were found in the area ventralis telencephali pars supracommissuralis dorsal to the anterior commissure. In the diencephalon, there was several GAL-ir perikarya in the nucleus preopticus periventricularis (NPP). Males had many GAL-ir perikarya in the nucleus preopticus pars parvocellularis (NPOpp) and isolated GAL-ir perikarya in the NPO pars magnocellularis, and lateral to the NPO; in females GAL-ir perikarya were not found in these sites. A large GAL-ir neuronal aggregation was observed in the nucleus lateralis tuberis pars posterioris (NLTp). Several ir perikarya were present in the nucleus posterioris tuberis; however, unlike in other regions the males revealed fewer neurons than females. Besides the established innervation of the pituitary gland by the NPP, NPO and NLT, the present study revealed GAL-ir perikarya of these nuclei apparently also innervating the telencephalon, thalamus, optic tectum, tegmentum and even some areas of the rhombencephalon. Isolated perikarya were found in the nucleus posterioris periventricularis, the dorsal vicinities of the nucleus recessus lateralis (NRL), nucleus recessus posterioris, and nucleus saccus vasculosus, and in the medulla oblongata ventral to the vagal lobes. In the pituitary gland, GAL-ir fibers ramify and terminate among the pars distalis cells. A small percentage of growth hormone-secreting cells colocalize GAL. In males, most GAL-ir cells of the proximal pars distalis (PPD) showed granular ir product in the entire cell, and some had one or two large granules; in females the ir PPD cells showed clusters of a few fine ir granules of uniform size in each. Sexual dimorphism was also found in the olfactory bulb, telencephalon, infundibulum, mesencephalic tegmentum, optic tectum and medulla oblongata, the males having a more extensive GAL-ir fiber system than the females. Galanin may play a role in both hypophysiotropic and motor functions.
Millions of women wear high heels on a daily basis; however, few studies have analyzed the changes high heels (positive heel inclination) have on posture. The purpose of this study was to determine whether positive heel inclination changed the postural alignment of the head, spine, pelvis, and knees. Fifteen female college students ((mean age = 22.7, SD = 3.7 years) had sagittal plane angles measured for the cervical spine, thoracic spine, lumbar spine, sacral spine, and knee joints in addition to anterior/posterior displacements of the head and pelvis. All variables were assessed by a Metrecom Skeletal Analysis System, a three-dimensional electrogoniometer. Six randomized trials, three at zero heel inclination and three at 5 cm positive heel inclination, were measured. Analysis of variance results indicated positive heel inclination of subjects brought about significantly lower anterior pelvic tilt, lumbar lordosis, and sacral base angles when compared with zero heel inclination (p < .01). Clinically, patients with low back pain may be affected by high heel usage because of the reduction of the normal lumbar lordosis.
In previous studies, we have demonstrated that inhibin and activin are stimulatory to goldfish gonadotropin release. In the present study, the distribution of immunoreactive inhibin and activin subunits (alpha, beta A, and beta B) in goldfish gonads was examined with domain-specific antibodies against mammalian inhibin and activin subunits, using the avidin-biotin-peroxidase complex method. In the ovary, follicle cells surrounding the oocyte were heavily stained with anti-porcine beta A and human beta B subunits. The cytoplasm of previtellogenic oocytes, at all stages of the ovarian cycle, also showed strong reactions with anti-beta A and beta B. As ovarian recrudescence progressed, oocytes that started to accumulate cortical vesicles gradually lost their immunoreactivity, with the reaction intensity inversely proportional to the amount of cortical vesicles in the oocyte; when oocytes became full of cortical vesicles, the oocytoplasm was no longer immunoreactive to anti-beta A and beta B. The staining with antiporcine alpha subunit was similar to that with anti-beta A and beta B but the reaction intensity was weaker than that with anti-beta A and beta B. In the testis, the mature sperm in the tubular lumen exhibited a strong immunoreaction to anti-alpha antibody, while the interstitial tissue was completely negative. However, the immunoreactivities with anti-beta A and beta B in the testis were exclusively restricted to the interstitial tissues. These results provide evidence for the presence of inhibin and activin-like molecules in the goldfish and reveal the distribution of these molecules in the goldfish gonads.
The NHS has been moving rapidly towards establishing clinical directorates in all specialties. It has been part of an attempt, along with the resource management initiative, to involve doctors in management, and to encourage them to take responsibility for the managerial and resource dimensions to clinical activity. Haydn Cook identifies a problem.
Ninety-six crossbred barrows were randomly allotted to five replications of four treatments to determine the effects of diets containing 20% extruded full-fat soybeans (FFS) or 4% butter (B) on the growth, composition, and sensory characteristics of finishing pigs. Pigs were housed five per pen, except for one replication that had four pigs per pen, in environmentally regulated barns. Pigs were given ad libitum access to a corn-soybean meal diet for 11 wk (control); a corn-extruded soybeans and soft wheat midds diet for 11 wk (FFS-11); a corn-soybean meal diet for 6 wk, changed to a corn-extruded soybeans and soft wheat midds diet for 5 wk (FFS-5); or a corn-soybean meal diet for 6 wk, changed to a corn-soybean meal, soft wheat midds, and 4% butter diet for 5 wk (B). Feed intake and weight gain were measured once every 2 wk. Pigs were slaughtered and carcass data were collected. Sensory characteristics (tenderness, juiciness, pork flavor intensity, off-flavor intensity, and overall acceptability), shear force, moisture, and fat content were determined for the longissimus muscle. Sensory characteristics (pork flavor, off-flavor, rubbery, cohesiveness, and juiciness) and 2- thiobarbituric acid values were determined for ground pork (30% fat) after 1, 4, and 7 d for control, FFS-11, and B treatments. No differences (P > .05) in ADG were observed between diets. Feed efficiency of the FFS-11 group was better (P < .05) than that of the control or B groups. No consistent differences were observed for carcass, sensory, or shear-force characteristics of the longissimus muscle or ground pork.(ABSTRACT TRUNCATED AT 250 WORDS)
Dispersed pituitary cells of the goldfish were incubated with biotinylated [D-Lys6, Pro9-N-ethylamide] salmon gonadotropin-releasing hormone (sGnRH-A) then avidingold (10 nm), and were fixed, embedded and sectioned. Cells were identified as gonadotrophs, somatotrophs, or prolactin cells using specific hormone antisera and protein-A gold (20 nm) as a marker. Attachment of the biotinylated sGnRH-A to the pituitary cell sections was determined by scanning cell surfaces for the smaller gold particles using the transmission electron microscope. Attachment was observed on gonadotrophs and somatotrophs, but was negligible on prolactin cells. Preincubation with unlabelled salmon gonadotropin-releasing hormone or chicken II gonadotropin-releasing hormone, or omission of the salmon gonadotropin-releasing hormone analog, prevented the reaction. The direct visualization of specific gonadotropin-releasing hormone receptors on gonadotrophs and somatotrophs supports the existence of direct stimulatory actions of gonadotropin-releasing hormone on gonadotropin and somatotropin release in gold-fish.
Central axons of sensory ganglion (SG) neurons of the Xenopus tail enter the spinal cord via the ventral roots and travel dorsally and rostrally following a diagonal course within the lateral marginal zone (LMZ) to reach the dorsolateral fasciculus (DLF) (Nordlander et al.: Brain Res., 440:391-395, 1988). Axons are dispersed as they cross the cord. At the DLF they turn and travel together rostrally, sharing the fascicle with axons of primary sensory neurons (Rohon-Beard cells) already present in the tract. In this paper we analyze the growth patterns of the central projections of SG axons in the tail by using HRP applied to proximal branches of tail spinal nerves. Growth cones of the diagonal route are variable in configuration, often bearing processes that spread within the LMZ. Once the DLF, growth cones change shape, becoming distinctly linear. While growth cones navigating the diagonal part of the route never contact or fasciculate with other diagonal SG axons, SG growth cones and axons of the DLF are more closely associated with their fellows. Measurements of the slopes of SG axons in the diagonal route indicated a limited range with a mean of 23 degrees with respect to the cord axis. On the basis of these observations, we conclude that 1) navigational patterns for growth cones of this pathway differ for the diagonal versus the DLF part of its course, and 2) fasciculation is not a mechanism used by SG axons to reach the DLF, but that instead, each axon is able to find its way independently.