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Biomedical subjects

D J Wilson

Publications and source records attributed to D J Wilson.

At least 163 records · Page 9Linked to original sources
PubMed1992

Preparation and characterization of type 1 astrocytes cultured from adult rat cortex, cerebellum, and striatum.

Astrocytes have been prepared from adult rat cortex, cerebellum, and striatum, using a modification of the McCarthy-DeVellis (J Cell Bio 85:890, 1980) method. The cultures consist of 99% type 1 polygonal astrocytes, which divide more slowly than cells from newborn animals. One day after preparing the cultures, 90% of the cells are glial fibrillary acidic protein (GFAP)-positive and 80% are vimentin-positive by immunohistochemical staining, suggesting that they are present de novo and not derived from precursor cells. The astrocytes from adult brain respond to an elevation of intracellular cyclic AMP, following treatment with forskolin, by becoming more stellate in shape and putting out fine ramified processes. They contain the same amount of GFAP per mg protein, measured by immunoblot, as cells from newborn animals. These cultures thus offer the possibility of comparing the biochemical properties of astrocytes derived from adult animals with those from newborn animals, or with cultures of reactive astrocytes isolated from lesioned brain.

Age Factors↗
PubMed1992

The effect of hypoxia and low pH on the cytotoxicity of chlorambucil.

Studies with mouse tumors have shown that the effectiveness of certain chemotherapeutic agents can be enhanced if they are used in appropriate combination with an anti-hypertensive drug such as hydralazine. This results in reduced tumor blood flow with, among other things, a consequent decrease in oxygenation and increase in acidity in the tumor tissue. The purpose of the present work was to determine to what extent hypoxia and low pH are involved in the mechanism of this effect for chlorambucil. V79-WNRE cells were exposed to various drug concentrations under aerobic or hypoxic conditions, pH 6.4 or 7.4. Measurements of cell survival following 1 hr exposure at 37 degrees C showed that pH 6.4 produced a large potentiation of cell killing by chlorambucil (ER = 4 approx.); hypoxia, on the other hand, had little effect. The potentiation was shown to be greatest for pH values below 7.0. HPLC measurements of drug uptake were made since it was anticipated that chlorambucil, a weak acid, might tend to accumulate in cells under conditions of low extracellular pH. It was found that at an extracellular pH of 6.4 the ratio of the intracellular (Ci) and extracellular (Ce) drug concentrations was increased 4.5 and 3.6 fold for aerobic and hypoxic conditions, respectively. This probably explains most, if not all, of the cell killing potentiation observed at low pH.

Animals↗
PubMed1992

Renal hemodynamics, urinary eicosanoids, and endothelin after liver transplantation.

Patients with hepatic cirrhosis develop widespread abnormalities in kidney function and vasoactive hormones. These change rapidly after liver transplantation during immunosuppression with cyclosporine. The role of changing eicosanoid excretion and endothelin levels in regulating renal function after transplantation in humans remains uncertain. We studied 32 patients with regard to renal hemodynamics, glomerular filtration, urinary prostacyclin (6-keto-PG-F1-alpha), thromboxane (TBX2), and endothelin before and during the first four weeks after orthotopic liver transplantation. Arterial pressure rose from 106 +/- 2/61 +/- 2 to 146 +/- 4/81 +/- 2 mmHg, (P less than .001), while renal blood flow fell (686 +/- 38 to 453 +/- 24 ml/min/1.73 m2, P less than .05), as did GFR. Pretransplant excretion of 6-keto and TBX2 was above that of normal subjects and fell progressively after transplant, as did plasma renin activity and aldosterone. The 6-keto levels fell below normal after two weeks. The ratio of TBX2/6-keto remained elevated compared with normal subjects throughout the month after transplant (1.54 +/- 0.38 vs. 0.54 +/- 0.07, P less than .01). Endothelin levels rose during the first week (7.4 +/- 1.4 vs. 12.4 +/- 2.7 pg/ml, P less than .05), but fell back to baseline thereafter. These results indicate that high levels of urinary eicosanoids in patients with liver disease fall rapidly after liver transplantation during CsA immunosuppression. Unlike results in many experimental models, these data suggest that renal vasoconstriction in humans may be associated primarily with suppression in renal prostacyclin excretion rather than stimulation of thromboxane.

Adult↗
PubMed1992

Culture of human osteoblasts on demineralised human bone. Possible means of graft enhancement.

We cultured human osteoblasts from trabecular bone explants and confirmed their phenotype by alkaline phosphatase assay, increased cyclic adenosine monophosphate production in response to prostaglandin E2 and radiographic micro-analysis of nodules of calcification. The osteoblasts were seeded on to demineralised human bone fragments and examined at ten-day intervals over a 50-day period by scanning electron microscopy. During this time the bank bone became progressively repopulated by the cultured osteoblasts. This system may offer a means of graft enhancement in elective orthopaedic and maxillofacial surgery by delivery of cultured autologous human osteoblasts to bone defects.

Alkaline Phosphatase↗
PubMed1992

Expression of growth factor mRNA in rabbit PVR model systems.

Proliferative vitreoretinopathy (PVR) involves the formation of intravitreal fibrocellular membranes which may lead to traction retinal detachment and blindness. The cellular component of epiretinal membranes originates from the proliferation and migration of cells within the eye. Several growth factors and other cytokines are plausible candidates for directing the processes leading to membrane formation. A reproducible animal model is needed for experimental studies of cytokine expression during PVR induction or treatment. We found that intravitreal injection of > 10(6) mixed mononuclear leukocytes or adherent monocytes along with a trans-scleral incision through the pars plana leads to the development of PVR-like disease in rabbit eyes. The severity of the disease was related to the number of monocytes injected. Typically, organized membranes extending from the incision toward the optic nerve formed within one week. Progression to extensive traction retinal detachment required 1 to 4 weeks. Injection of up to 5 x 10(6) lymphocytes or freeze-thaw killed monocytes was ineffective, and coinjecting 100 micrograms endotoxin with the monocytes did not result in enhanced disease. The histological appearance of the epiretinal membranes was similar to human PVR membranes. Macrophage, cytokeratin-positive (epithelial), and fibroblast-like cells were present. Northern blot analysis of RNA extracted from the rabbit membranes revealed the presence of mRNA for acidic fibroblast growth factor (aFGF). Acidic FGF mRNA was not expressed by the injected monocytes. A comparable level of aFGF mRNA and also mRNAs for basic FGF, platelet-derived growth factor-B, and transforming growth factor beta were found in epiretinal membranes induced by a scleral incision in association with cryopexy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Detection of antibiotic and sulfonamide residues in bob veal calf tissues: 967 cases (1987-1988).

The data on antimicrobials in bob veal calves obtained by USDA-Food Safety Inspection Service (FSIS) during the fiscal year 1988 were reviewed. Of 3,095 calf antibiotic and sulfonamide test (CAST)-positive carcass submissions, 967 were chosen by FSIS for antimicrobial residue analyses. Specimens of muscle, liver, and kidney were obtained at abattoirs as a part of the FSIS program, and the results were reviewed by calf submission. At least 1 tissue from each of 425 submissions contained neomycin. Unidentified microbial inhibitors were found in 291 submissions. Streptomycin was found in 86 carcass submissions, penicillin in 81, sulfamethazine in 73, tetracycline HCl in 59, and gentamicin in 53. Other monitored agents were found in less than 50 submissions each. Only 257 submissions included evaluation for sulfonamides, but sulfamethazine was found in 28.4% of them. Chloramphenicol was not detected. Concentrations of neomycin in kidney specimens ranged from 0.25 to greater than 100 ppm, and differed among the 3 regional laboratories (West, Midwest, and East). The western region had the greatest proportion of low-concentration specimens, whereas the midwestern laboratory had the largest proportion of specimens with very high concentrations (greater than or equal to 100 ppm) of neomycin in kidney tissue. Data to identify the sources and causes of the residues were not available. However, the western laboratory evaluated CAST-positive submissions from calves certified to be antimicrobial-free, whereas the other laboratories evaluated submissions from certified and noncertified calves; therefore, this may partially explain regional patterns for neomycin quantities in kidney specimens. Neomycin predominated among CAST-positive verifications. A high prevalence of unidentified microbial inhibitors suggests further identification may be warranted.

Abattoirs↗
PubMed1991

Evidence for isoleucine as a positive effector of the ilvBN operon in Salmonella typhimurium.

Concerted efforts were directed towards understanding the control of acetohydroxy acid synthase (AHAS) in the gyrB mutant hisU1820 of Salmonella typhimurium. A media shift from valine to valine plus isoleucine causes a dramatic 4 to 5 fold burst of AHAS valine sensitive activity which appears to be dependent on translation. DJ19, an isolated valine sensitive derivative of the gyrB mutant, maintains a dramatic increase in AHAS valine sensitive activity upon the addition of isoleucine to valine supplemented cultures, suggesting that the isoleucine effect is specific for valine sensitive AHAS. Evidence supports isoleucine as a positive effector on valine sensitive AHAS expression and that the gyrB mutation accentuates the isoleucine effect.

Acetolactate Synthase↗
PubMed1991

Antibiotic and sulfonamide residues from Food Safety Inspection Service bob veal calf tissues by region, from October 1987 to September 1988.

Of 3,095 bob veal carcasses declared positive to the calf antibiotic and sulfonamide test (CAST) performed by USDA-FSIS personnel during the fiscal year 1988, 967 were submitted to USDA-FSIS laboratories for intensified antimicrobial evaluation of kidney, liver, and muscle specimens. In the western region, CAST-positive specimens were analyzed from certified calves, that is, calves declared free of antimicrobials by their producers. In the midwestern and eastern regions, 20% of the CAST-positive carcass submissions were certified and 20% had not been certified. Among the regions, neomycin was the most commonly reported agent from kidney specimens; however, there were some distinct differences among regions (24.6% of the specimens in the Midwest, 37.4% in the East, and 46.8% in the West). Reports of gentamicin in specimens also were different among regional laboratories (9.0% in the Midwest, 12.1% in the East, 3.1% in the West). Sulfamethazine was less frequently reported by the western laboratory, never exceeding 2% among the 3 tissues tested, whereas the eastern and midwestern laboratories had at least 1 tissue type that was greater than or equal to 9%. Not all specimens were subjected to sulfonamide evaluation. Regional patterns were noticed for unidentified microbial inhibitors, which were most evident for liver in which 24.6% of the specimens at the western laboratory were classified as having unidentified microbial inhibitors, and only 6.3% of specimens at the eastern laboratory were classified as having unidentified microbial inhibitors.

Animals↗
PubMed1991

Full length mouse glycophorin gene constructed using recombinant polymerase chain reaction.

Recently, an incomplete cDNA clone for a major mouse glycophorin gene, pGP315, and a genomic clone, pGX7 (which contains the first exon and nucleotide sequences around the transcription start sites) was isolated and sequenced by Matsui et al. (1). Since there were no available restriction sites for the construction of a full length mouse glycophorin A gene, the recombinant PCR technique was adapted to splice together the above two partial sequence clone inserts to obtain a full length recombinant DNA fragment (1053 bp) containing the proper sequence of mouse glycophorin A cDNA. The PCR reconstructed DNA fragments were verified by: gel electrophoresis to contain the expected sizes, hybridization to probes made from the DNA components before recombination, and confirmed by the restoration of a previously destroyed restriction enzyme site. The corrected gene sequence for pGP315 is also reported.

Animals↗
PubMed1991

Species specificity in mouse glycophorin.

89, 65, 46 and 29 Kd mouse glycophorin proteins identified during polyacrylamide electrophoresis of mouse erythrocytes have been further characterized. These proteins (1) stain positive with Periodic Acid Schiff reagent after sodium hydroxide treatment; (2) labeled using [125I] in intact cells; (3) co-isolated along with integral membrane proteins in the pellet fraction of sodium hydroxide treated ghosts; and (4) demonstrated a molecular weight downshift after neuraminidase treatment during electrophoresis. We have called them mouse Sialoglycoproteins 1, 2, 3 and 4. Immuno-blot analysis revealed distinct species specificity between human and mouse erythrocyte ghosts, and some cross-reactivity between rat and mouse erythrocyte ghosts.

Animals↗
PubMed1991

Ocular clinicopathologic study of gyrate atrophy.

We performed a histopathologic study of whole globes obtained post mortem from a patient with well-documented, vitamin B6-responsive gyrate atrophy. The retina in the posterior pole had focal areas of photoreceptor atrophy with adjacent retinal pigment epithelial hyperplasia. An abrupt transition from the near normal retina to a zone of near total atrophy of the retina, retinal pigment epithelium, and choroid was present in the fundus midperiphery. Electron microscopic examination disclosed abnormalities of the mitochondria of the corneal endothelium and the non-pigmented ciliary epithelium. Similar, but less severe, mitochondrial abnormalities were present in the photoreceptors.

Aged↗
PubMed1991

Chick frontonasal process excision significantly affects mid-facial development.

The midfacial region in vertebrates may be considered as developing from five separate processes, namely the central frontonasal process (FNP) and the paired maxillary and lateral nasal processes. Relatively little is known about the mechanisms whereby these processes interact to produce structures of the neonatal/adult face. This study was undertaken to gain some insights into the events involved in this process, and involved observing the effects on facial development in the chick of surgical excision of the FNP, prior to its fusion with the other facial processes. In the absence of the FNP, outgrowth of the upper beak was dramatically reduced, agenesis of the primary palate occurred, and development of the maxillary processes and palatal shelves was impaired. Thus, in the chick, the frontonasal process plays a major role in midfacial morphogenesis. Not only does the FNP provide the primary palate and a contribution to the development of the nasal septum, it is also important in the ordered development of the maxillary processes and of the definitive secondary palate--contributions which have not emerged clearly from in vitro and teratogenic studies.

Animals↗
PubMed1991

Isolation of murine sialoglycoprotein using consecutive chromatography.

Affinity columns and high performance liquid chromatography were employed consecutively to obtain 89, 65, 46 and 29 kilodalton sialoglycoproteins from mouse erythrocyte ghosts free of the Band 3 protein which traditionally co-purifies with these proteins. The purification scheme involves Concanavalin A, Wheat Germ Agglutinin and/or Limulus lectin Sepharose 4B columns. We have designated these glycophorin-like proteins Sialoglycoproteins 1, 2, 3, and 4, respectively. Sialoglycoprotein 2 can be isolated independently using a Limulus column combination, while Sialoglycoproteins 3 and 4 were isolated separately during high performance liquid chromatography, demonstrating heterogeneity in binding properties between these sialoglycoproteins.

Animals↗
PubMed1991

Synthetic murine epidermal growth factor sequence 20-31 is mitogenic and angiogenic.

Epidermal growth factor (EGF) and its homologue, transforming growth factor alpha (TGF alpha), are mitogenic, angiogenic and tumour-promoting polypeptides. Much effort has therefore been directed towards the development of EGF/TGF alpha antagonists as a potential cancer therapy. Initial reports that some EGF/TGF alpha synthetic fragments possess EGF-receptor binding activity have not been confirmed in subsequent studies. We have found, however, that the murine EGF B-loop sequence: Ac-[(S-acetamidomethyl)-Cys20,31]-EGF-(20-31)-NH2 [(mEGF-(20-31)] produces biological effects consistent with the parent molecule in bovine, murine, chick and human, but not rat, model systems. In parallel experiments, both mEGF and mEGF-(20-31) elicit migratory, cytoprotective, growth-stimulatory, growth-inhibitory and angiogenic responses. The reverse B-loop sequence, mEGF-(31-20), is also mitogenic and angiogenic. The C-loop sequence, mEGF-(33-42), has no mitogenic or angiogenic activity when applied alone, does not block the mitogenic effect of mEGF, but does block the angiogenic effect of mEGF. It has not been established that the EGF receptor is the target for these fragments, but the results suggest that the residual biological activities of EGF fragments merit further investigation.

3T3 Cells↗
PubMed1991

Transconjunctival frontalis suspension (TCFS).

We have devised a frontalis suspension technique originally described in an article entitled, "Transconjunctival frontalis suspension for blepharoptosis" (Trans Am Acad Ophthal Otolaryngol 83:684-92), by Swan and Tongue in 1977. This modified transconjunctival frontalis suspension (TCFS) procedure is described. It has been used at the Oregon Health Sciences University in its original or modified form since 1970. To better understand how this procedure successfully corrects blepharoptosis in patients with poor levator function, a study was undertaken to discern the anatomic relationship of the suspension material to important ocular adnexal structures. The findings are based on gross anatomic dissection and histologic examination of a cadaver eyelid after postmortem transconjunctival fascia lata frontalis suspension. The distal, horizontal limb of the fascia suspends the central portion of the levator aponeurosis in a hammock-type fashion. The two vertical limbs pierce the aponeurosis at the level of Whitnall's ligament and incorporate this ligament. The fascia then exits the orbit through the orbital septum near the arcus marginalis and is incorporated into the frontalis muscle near the junction of the medial one-third and lateral two-thirds of the brow, temporal to the supraorbital neurovascular bundle. This anatomic location is not only relatively easily and safely attained, but also maximizes the principles of more traditional frontalis suspensions while minimizing the effort and surgical manipulation.

Blepharoptosis↗
PubMed1991

N-acetyl-beta-D-glucosaminidase, etiologic agent, and duration of clinical signs for sequential episodes of chronic clinical mastitis in dairy cows.

This study examined effects of repeated episodes of clinical mastitis in chronically infected quarters on milk N-acetyl-beta-D-glucosaminidase activity and duration of clinical signs. Milk samples were collected at each clinical onset from 49 chronic mastitis cases on a 1700-cow Michigan dairy farm. There were 49 first episodes of clinical mastitis, 49 second episodes, and 13 episodes of third or more. Agents isolated were Staphylococcus aureus (18.4%), Staphylococcus (7.3%), no growth (20.2%), environmental pathogens (streptococci other than agalactiae, Escherichia coli, Klebsiella spp., Enterobacter spp., and Citrobacter spp.) (22.0%), other pathogens (Serratia spp., Bacillus spp., diphtheroids [Corynebacterium spp. and Actinomyces pyogenes], Pseudomonas spp., and Nocardia spp.) (11.9%), mixed pathogens (two agents isolated) (12.8%), and contaminated samples (7.3%). Etiologic agents, duration of clinical signs, and NAGase did not differ by episode number. The correlation between log of NAGase and log of time until clinical recovery was .34. The relationship between NAGase and duration of clinical signs was strongest for second episodes, and weakest for third and greater episodes of chronic mastitis.

Acetylglucosaminidase↗
PubMed1991

Temporal patterns of lost milk production following clinical mastitis in a large Michigan Holstein herd.

Cows with three hundred and sixteen cases of clinical mastitis were sampled by microbiologic culture during a 6-mo period on a 1700-cow Michigan Holstein dairy farm. Daily milk weights were obtained on all cows before clinical onset and for 60 d after onset. Predicted post-mastitis production, projected on the basis of premastitis production and the lactation curves of contemporary non-mastitic herdmates, was compared with actual daily milk production during the 60 d following clinical onset. Cows experiencing clinical mastitis produced approximately 341 kg less salable milk during the 60 d after clinical onset compared with projected production. This milk loss included both decreased production and milk withheld from market following antibiotic treatment. Pluriparous cows lost 2.06 times as much milk as first lactation cows, and cows with mastitis occurring before 150 d in lactation lost 1.40 times as much milk when compared with other cows. Cows with mastitis occurring in the winter showed a milk loss 1.37 times greater than cows with mastitis in summer. The identity of the mastitis agent isolated from the clinical case was not strongly associated with the drop in milk production in the 60 d following clinical onset.

Animals↗