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Biomedical subjects

W L Davis

Publications and source records attributed to W L Davis.

At least 55 records · Page 3Linked to original sources

The electron microscopic immunolocalization of a copper-zinc superoxide dismutase in association with collagen fibers of periodontal soft tissues.

Periodontal soft tissues were cleaved from freshly extracted human teeth. Tissues were then prepared for subsequent biochemical and morphological studies according to the following plan: 1) immediate immersion in liquid nitrogen for the biochemical assay of superoxide dismutase (SOD); 2) immediate fixation prior to routine preparation for routine transmission electron microscopy; 3) immediate fixation prior to preparation for electron microscopic immunohistochemistry. Biochemical analysis showed that the human periodontal ligament contained about twice as much SOD activity as human skin (dermis), but considerably less enzyme activity than that seen in red blood cells. Interestingly, periodontal SOD activity appeared to decrease with age. Immunohistochemistry localized enzyme activity to the periphery of matrix collagen fibrils and to the glycocalyx of tissue fibroblasts. The pathophysiology of this enzyme regarding inflammatory diseases such as periodontitis is discussed.

Adult↗

The hospital of the future--planning for change.

This paper describes the hospital planning model developed by the North Eastern Metropolitan Region of the Health Department Victoria to forecast acute public hospital bed-day requirements in the Region. Three age-specific variables: population; separation rate; and length of stay have been used to estimate the level of demand for hospital services. The model also delineates services delivered on a same day or long stay basis. The application of the model to three local government areas demonstrates the importance of population growth and ageing on the type and level of hospital services required and the implications thereof for service delivery and the physical configuration of hospitals.

Age Factors↗

The major obstructive inflammatory patterns of the sinonasal region seen on screening sinus computed tomography.

In conclusion, inflammatory sinonasal disease can be conveniently grouped into five distinct radiological patterns, each with a different therapeutic course and surgical options. A more precise interpretation of SSCT scans is rendered when inflammatory sinonasal disease is categorized into these distinct radiological patterns. The three obstructive patterns occur due to dysfunction of the mucociliary drainage routes of the paranasal sinuses and result in specific diagnostic patterns that are recognizable on coronal SSCT examinations. When one of the three obstructive patterns is identified, detailed attention can be directed to the likely site of occlusion, with possible definition of a specific etiology. A detailed road map of relevant surgical anatomy and pathology is then available for the endoscopic surgeon. This road map can then be used for a more directed and specific functional endoscopic sinonasal surgery. This results in improved patient care and surgical result.

Humans↗

Immunohistochemical and pharmacokinetic characterization of the site-specific immunoconjugate CYT-356 derived from antiprostate monoclonal antibody 7E11-C5.

In this study, a site-specific immunoconjugate, designated CYT-356, of the prostate-reactive monoclonal antibody 7E11-C5 was characterized by immunohistological methods for reactivity with normal and neoplastic human tissues. In addition, CYT-356 labeled with 111In was assessed by in vivo imaging and pharmacokinetic studies for localization to human tumor xenografts in nude mice. The native antibody and the site-specific immunoconjugate exhibited similar patterns of reactivity with normal human tissues. Although the majority of tissues tested were negative, weak reactivity with cardiac muscle, proximal kidney tubules, and sweat glands was observed. Positive staining of normal prostate epithelial cells and glandular lumina and strong reactivity with a subset of skeletal muscle cells were also observed. CYT-356 reacted with 100% of prostate tumors examined but was negative on a variety of other neoplasms. Following i.v. administration, CYT-356-111In rapidly localized to and imaged LNCaP human prostate adenocarcinoma xenografts in nude mice, reaching maximal levels of about 30% of injected dose/g of tumor within 3 days. No unusual localization was seen to any nontumor tissue or organ; the level of radioactivity in the normal tissues and organs was at or below that seen in the blood. The localization to xenografts was antigen specific and the accessible binding sites in 100-200-mg tumors appeared to be saturated at an antibody dose between 10 and 100 micrograms. These findings suggest that the CYT-356 immunoconjugate may be useful in the diagnosis and therapy of prostate cancer.

Animals↗

Hibernation activates glyoxylate cycle and gluconeogenesis in black bear brown adipose tissue.

Biochemical studies on brown adipose tissue removed from a hibernating black bear and a non-hibernating control animal demonstrate that this tissue: (1) can carry out cyanide-insensitive fatty acid oxidation, and (2) possesses catalase activity and the enzyme activities unique to the glyoxylate cycle, isocitrate lyase and malate synthase. These activities are all markedly increased in brown fat obtained from the hibernating animal. Additionally, hibernation enhances the ability of the tissue to synthesize glycogen in the presence of a fatty acid substrate. The glyoxylate cycle enzymes and the ability to convert fatty acid carbons to glucose have been generally regarded as being absent from vertebrate cells and tissues.

Adipose Tissue, Brown↗

Identification of glyoxylate cycle enzymes in chick liver--the effect of vitamin D3: cytochemistry and biochemistry.

Information regarding the presence of the glyoxylate cycle in chick liver was sought. This metabolic pathway has long been thought to be absent from vertebrate tissues. Previous studies in other tissues have shown that, when present, this pathway is sensitive to vitamin-D. Thus, the effect of long-term vitamin-D deficiency and subsequent vitamin-D replacement on liver structure was studied by light microscopy. In addition, specific biochemical assays for the presence of glyoxylate cycle enzymes were performed. Light microscopy of lipid extracted tissues, light microscopic histochemistry, and quantitative histochemistry showed that the hepatocytes from vitamin-D-deficient animals contained primarily lipid. Hepatocytes from normal and vitamin-D-replete livers contained primarily carbohydrate as judged by their staining with periodic acid-Schiff (PAS). Also, malate synthase positive peroxisomes were seen in hepatocytes from normal and vitamin-D-treated chicks. Structures positive for this glyoxylate cycle enzyme were rarely seen in the hepatocytes from vitamin-D-deficient animals. Biochemical analyses showed the presence of the two unique glyoxylate cycle enzymes, isocitrate lyase and malate synthase, in chick hepatocytes. The activity of these enzymes was markedly increased in the vitamin-D-replete livers. In addition, chick hepatocytes demonstrated the capacity to oxidize fatty acid in the presence of cyanide. This activity, which is characteristic of peroxisomal B-oxidation rather than mitochondrial, was stimulated by vitamin-D treatment. Lastly, incubation of chick liver in the presence of a fatty acid substrate (palmitate) led to higher tissue glycogen content. The latter was further increased in liver from vitamin-D-replete animals. These data show the presence of glyoxylate cycle enzymes in a higher vertebrate and indicate that this tissue is endowed with the capacity to convert lipid to carbohydrate.

Animals↗

Carbohydrate-derivatized immunoconjugate of the anti-(carcinoembryonic antigen) monoclonal antibody C46: immunohistological reactivity and pharmacokinetic comparison with a randomly derivatized C46 immunoconjugate.

In this study, a site-specific glycyl-tyrosyl-(N-epsilon-diethylenetriaminepentaacetic acid)-lysine (GYK-DTPA) immunoconjugate of the anti-carcinoembryonic antigen monoclonal antibody C46 (C46-GYK-DTPA) was characterized by immunohistological and immunofluorescence methods for reactivity with normal and neoplastic human tissues. In addition, pharmacokinetic studies assessed the ability of C46-GYK-DTPA labeled with 111In to localize to and image human tumor xenografts in nude mice. The native antibody and the site-specific immunoconjugate exhibited similar patterns of reactivity with normal human tissues. C46 did not bind to the surface of normal human granulocytes, which indicates lack of reactivity with normal cross-reacting antigen. C46-GYK-DTPA reacted with 100% of the colon, breast and renal carcinomas examined and with two of three lung carcinomas, but did not react with any sarcomas, melanomas or lymphomas examined. Intravenously administered C46-GYK-DTPA-111In rapidly localized to and imaged LS174T human colon adenocarcinoma xenografts in nude mice, reaching maximal levels of about 25% of injected dose/g tumor within 1 day. No unusual localization to any non-tumor tissue or organ was seen; the level of radioactivity in the normal tissues and organs was at or below that in the blood. The accessible binding sites in 1 g tumors appeared to be saturated at an antibody dose between 100 micrograms and 1000 micrograms/mouse. Further, in a direct in vivo comparison, the site-specific conjugate C46-GYK-DTPA had more favorable pharmacokinetics and better tumor localization than a randomly derivatized C46 immunoconjugate (C46-DTPA). These findings suggest that the site-specific immunoconjugate C46-GYK-DTPA may be useful in the diagnosis and therapy of colon cancer and other adenocarcinomas expressing carcinoembryonic antigen.

Animals↗

Immunolocalization of secretory protein-I or chromogranin A in amphibian urinary bladder granular cell granules.

The presence of secretory protein-I (SP-I) or chromogranin A (CGA) in granules isolated from the granular cells of the amphibian urinary bladder epithelium was investigated using ultraimmunohistochemistry. Granules were isolated by cell fractionation using Percoll density gradients. SP-I was isolated and purified from bovine parathyroid glands. Antibodies were raised in rabbits and purified by affinity chromatography. Ultraimmunocytochemistry, employing the avidin-biotin-peroxidase (ABC-complex) procedure, was used to localize SP-I on thin sections of isolated granules. About 27% of the granules from control (-ADH) cells were SP-I+, while 51% of the granules fractionated from hormone treated (+ADH) cells were positive for this protein (p less than 0.0001). Accordingly, granules from ADH-treated cells also showed a significant (p less than 0.0001) increase in total protein.

Animals↗

Retropharyngeal space: evaluation of normal anatomy and diseases with CT and MR imaging.

A retrospective analysis of computed tomographic (CT) and magnetic resonance (MR) images and clinical records of 39 patients with retropharyngeal space (RPS) lesions was completed. The review was undertaken to answer the following questions: (a) what is the spectrum of lesions of the RPS? (b) what imaging features mark a lesion as originating in the RPS? and (c) is there a difference between the radiologic pattern of the suprahyoid and infrahyoid portions of the neck? Of the 39 patients in the study, nine had RPS infections, 17 had RPS malignancies, two had benign tumors, seven had RPS pseudotumors, and four had hematoma or air in the RPS after trauma. RPS lesions demonstrated two distinct radiologic patterns: a nodal pattern and a nonnodal pattern. The nodal pattern, found only in the suprahyoid neck, occurs when infection or tumor begins in the lymph nodes of the RPS. The lesions may be unilateral or bilateral, but the middle part of the RPS is spared. The nonnodal pattern, found primarily in the infrahyoid RPS, results when the infection or tumor directly invades the RPS or goes beyond the nodes of the RPS. The nonnodal lesion appears rectangular and spans the RPS from side to side.

Carcinoma, Squamous Cell↗

Ethanol induces the generation of reactive free radicals by neural crest cells in vitro.

Developmental craniofacial anomalies related to the neural crest derived ectomesenchymal cell population are associated with fetal alcohol syndrome (FAS). Information regarding any potential relationship between ethanol, free radicals, and the viability, proliferation, etc., of isolated neural crest cells was sought. The hypersensitivity of neural crest cells to ethanol was observed. This drug severely depressed cell viability while simultaneously inducing the generation of such reactive oxygen intermediates (ROI) as superoxide, hydrogen peroxide, and hydroxyl anions. Addition of the free radical scavenging enzyme superoxide dismutase to the culture medium significantly reversed these effects of ethanol. The cytotoxicity of ethanol was further confirmed by the release of radiolabeled chromium (51Cr) from cells prelabeled prior to ethanol treatment. This effect was also depressed by the addition of superoxide dismutase. Interestingly, an assay for superoxide dismutase activity showed that neural crest cells may be devoid of this enzyme. The latter may help to explain the overt sensitivity of these cells to such a broad spectrum of teratogens, many of which can either dissociate directly into ROI, or cause the radicalization of biological structures and molecules. Plasmalemmal lipids (via lipid peroxidation) and DNA are at an especially high risk from uncontrolled ROI. Changes in neural crest cell surface morphology, i.e., loss of microvilli, formation of xeiotic blebs, as well as the "leakage" of radiolabeled Cr from prelabeled cells, would seem to show that ethanol, as a result of induced free radical formation, alters the physiology and biochemistry of the cell membrane. These findings however, should not exclude other potential sites for ETOH-induced cell injury related to free radicals, especially the nuclei (DNA), mitochondria, organelle membranes, and the cytoskeleton.

Animals↗

Generation of radical oxygen species by neural crest cells treated in vitro with isotretinoin and 4-oxo-isotretinoin.

The effects of isotretinoin (IR) and its primary metabolite (in the human), 4-oxo-isotretinoin (4-OIR or OIR), on isolated chick neural crest cells (NCC's) in culture were studied. NCC's were found to be deficient in both superoxide dismutase (SOD) and catalase, two of the enzymes known to function in the "scavenging" (dismutation) of toxic radical oxygen species (ROS) such as the superoxide anion and hydrogen peroxide. The addition of IR or OIR to the culture medium significantly depressed the viability of the NCC's when compared to untreated cells. OIR was more potent in this regard than IR. In the presence of either IR or OIR, NCC's generated superoxide anions (O2.), hydrogen peroxide (H2O2), and hydroxyl anions (OH.). OIR was again more potent. The cytotoxicity of IR or OIR was demonstrated by the "leakage" of radioactive chromium from prelabeled cells. The latter is suggestive of a primary surface membrane defect, most logically via the induction of lipid peroxides by the retinoids. The latter is accompanied by an increase in membrane permeability and porosity as evidenced by the fact that various fluorescently labeled molecules, including BSA-FITC (MW 69,000), gain entrance into the cytoplasm of the retinoid treated cells. No label was seen in the cytoplasm of similarly treated control cells. When SOD (200 units/ml) or catalase (400 units/ml) was added to the culture media of IR- or OIR-treated NCCs, cell viability was increased and the concentration of the various ROS generated was decreased. Membrane leakiness to chromium and FITC-BSA was also decreased in the presence of these enzymes. Free radicals, when not inactivated (dismutated), are known to be pathobiotic to most cells. Cell membranes are at a particular high risk from ROS which induce structural, physiological, and biochemical alterations in the cell membrane. The latter can have a negative effect on cell permeability, maintenance of normal ionic gradients, membrane enzyme activity, cell-to-cell communication, etc. Such defects can ultimately culminate in hypoplasia, aplasia, and cell necrosis. This study has shown that NCC's may be overtly sensitive to ROS, especially since these undifferentiated cells apparently lack inherent SOD and/or catalase activity. From this study it appears as if both IR and OIR perturb the normal functional state of NCC's by "triggering" the generation of ROS. This may certainly explain the teratogenicity of these drugs as related to the viability of neural crest derived ectomesenchymal cells and normal craniofacial morphogenesis.

Animals↗

Glyoxylate cycle in the epiphyseal growth plate: isocitrate lyase and malate synthase identified in mammalian cartilage.

Peroxisomes were identified in chondrocytes from all zones of the mammalian epiphyseal growth plate by using light microscopic techniques for the cytochemical demonstration of catalase, the marker enzyme for these organelles. Additional cytochemistry showed the presence of malate-synthase-positive structures within the chondrocytes. The latter enzyme, also associated with peroxisomes, is unique to the glyoxylate shunt, a metabolic pathway thought to be absent in vertebrate tissues. The glyoxylate cycle allows the net conversion of lipid to carbohydrate, i.e., gluconeogenesis. Biochemical studies on growth plate cartilage indicate that this tissue has the capacity to carry out cyanide-insensitive B-oxidation of fatty acids. The latter takes place in a nonmitochondrial compartment, most likely the peroxisomal compartment. Additionally, both of the unique enzymes associated with the glyoxylate cycle, i.e., isocitrate lyase and malate synthase, were also identified in a cell-free homogenate of this cartilage. These studies indicate that cartilage, a poorly vascularized tissue characterized by its low oxygen tension and anaerobic glycolysis, may have the capacity to convert lipid to carbohydrate, i.e., gluconeogenesis via the glyoxylate pathway. In this way, cartilage may be unique among mammalian tissues.

Animals↗

Insect hemolymph factor promotes muscle histolysis in Solenopsis.

Hemolymph was collected from both normal (virgin) females (control hemolymph) and artificially inseminated females (experimental hemolymph) of Solenopsis, ssp., the imported fire ant (primarily S. invicta and S. geminata). When the control hemolymph was injected into normal females, breakdown of the thoracic flight musculature was not seen 24 hr postinjection. In contrast, when the experimental hemolymph was injected into normal females, flight muscle histolysis was marked 24 hr postinjection. When the experimental hemolymph was heated to 70 degrees C prior to injection into normal females, subsequent flight muscle breakdown was not seen. The injection of freshly collected semen into normal females produced no effect on flight muscle structure. Also, the injection of the experimental hemolymph into normal, alate males produced no muscle histolysis. These observations suggest that the hemolymph from inseminated females contains a factor (or factors) that induces the specific breakdown and subsequent dissolution of the complex thoracic flight musculature. Other thoracic muscles (leg muscles, intersegmental muscles, etc.) are not affected. Based on observations made in both insect muscles as well as vertebrate skeletal muscles, it is suggested that such a hemolymph factor may act by disrupting the structural integrity of muscle cell membranes, resulting in significant changes in membrane permeability, especially with regard to calcium ions.

Animals↗

The immunocytochemical localization of superoxide dismutase in the enterocytes of the avian intestine: the effect of vitamin D3.

Both light microscopical and electron microscopical immunocytochemical techniques were utilized to localize CuZn-superoxide dismutase (SOD) in the duodenum of normal, rachitic and vitamin-D3-replete chicks. This enzyme catalyses the dismutation of the superoxide anion, a toxic free radical generated during the normal aerobic metabolism of most respiring cells. Light microscopy showed no SOD activity associated with the duodenal enterocytes of normal and rachitic chicks. However, in rachitic animals subsequently treated with vitamin D, i.e. vitamin-D-replete chicks, intense immunoreactivity for the enzyme was seen in association with the apical border of the duodenal absorptive cells. Immunostaining for SOD was not seen in goblet cells. With electron microscopy, immunostaining for SOD activity was identified in association with the apical microvilli and, to a lesser degree, with the terminal web, a well as in association with both lysosomes and peroxisomes. From this report it appears that there is a physiological relationship between vitamin D, SOD and the intestinal absorptive cell. However, the precise relationship must await further clarification.

Animals↗

The ultrastructural immunocytochemical localization of superoxide dismutase in the amphibian urinary bladder: effect of aldosterone.

Transmission electron microscopy and immunohistochemistry, the latter employing the avidin-biotin-peroxidase (ABC complex) technique, were utilized to localize copper-zinc superoxide dismutase (CuZn-SOD) enzyme activity in the epithelial cells of the toad urinary bladder mucosa. This 'scavenger' enzyme catalyses the dismutation (reduction-oxidation) of the superoxide anion (O2-), a toxic free radical generated during normal cellular respiration. In unstimulated epithelial cells, enzyme activity was seen in the cytosol of granular, mitochondrial-rich and goblet cells. The basal cells were generally devoid of enzyme activity. In addition to the cytosol, SOD activity was also seen in association with the apical plasma membrane of the epithelial cells. In the presence of the steroid hormone aldosterone (10(-7)M, 30 min-6h), CuZn-SOD activity was markedly increased along the luminal mucosal membrane of granular, mitochondrial-rich and goblet cells. This increase was seen as early as 30 min after the addition of hormone, and as long as 6h after treatment. The cytosolic reaction was usually decreased or absent under these conditions. From the data presented, it appears that CuZn-SOD is involved in electrolyte (sodium) transport in the epithelial cells of the toad urinary bladder. The latter may involve hormone-induced alterations in luminal cell membrane structure and chemistry.

Aldosterone↗

The immunohistochemical localization of superoxide dismutase activity in the avian epithelial growth plate.

Superoxide dismutase (SOD) is a 'scavenger' enzyme which catalyses the dismutation (reduction-oxidation) of the superoxide anion (O2-.), a toxic free radical generated during normal cellular respiration. Light microscopy employing immunohistochemistry was utilized for localizing SOD activity in the chick epiphyseal cartilage. Antibodies to mammalian liver CuZn-SOD were prepared and the avidin-biotin-peroxidase technique (ABC complex) was utilized to localize activity for this enzyme in the growth plate cartilage. The localization of enzyme activity varied in accordance with the characteristic zonation pattern of the growth plate (zone of proliferation, zone of maturation, zone of cell hypertrophy and zone of matrix calcification). In the upper regions of the epiphyseal cartilage (the zones of proliferation and maturation), where the vascularity is poor and the oxygen tension low, SOD activity was localized within the chondrocytes. No extracellular activity was observed. However, in the lower regions of the growth plate (the zones of cell hypertrophy and matrix calcification), where both the vascularity and the oxygen tensions are increased, SOD activity was intense in both the chondrocytes and the surrounding extracellular matrix. Thus, the distribution of SOD enzyme activity in this tissue seems to vary in accordance with the level of oxygen present. The significance of the extracellular SOD activity, seen in the lower aspects of the growth plate cartilage, may indicate the sensitivity of matrix components, especially collagen, to toxic free radicals such as the superoxide anion.

Animals↗