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Biomedical subjects

J L Bryant

Publications and source records attributed to J L Bryant.

At least 37 records · Page 2Linked to original sources

Reduction of nitric oxide synthase activity in human neutrophils by oxidized low-density lipoproteins. Reversal of the effect of oxidized low-density lipoproteins by high-density lipoproteins and L-arginine.

Oxidized low-density lipoproteins (ox-LDL) inhibit vascular relaxation by decreasing the synthesis or rapid degradation of endothelium-derived relaxing factor (EDRF), now identified to be nitric oxide (NO). We examined the regulation of NO synthase activity in human neutrophils, which also generate NO, by lipoproteins. Isolated human neutrophils were incubated with native-LDL, ox-LDL (10-50 micrograms protein/mL), high-density lipoproteins (HDL, 100 micrograms protein/mL) or HDL+ox-LDL, and NO synthase activity was measured as conversion of [3H]L-arginine to [3H]L-citrulline. Ox-LDL, but not native-LDL or HDL, significantly decreased NO synthase activity in human neutrophils. This effect of ox-LDL was incubation time and concentration dependent. The incubation of cells with HDL or L-arginine diminished the effects of ox-LDL on NO synthase activity. Thus, ox-LDL decreases the activity of NO synthase enzyme, and this effect of ox-LDL can be modified by HDL and L-arginine.

Arginine↗

Systemic gene therapy: biodistribution and long-term expression of a transgene in mice.

We have investigated the in vivo efficacy of a systemic gene transfer method, which combines a liposomal delivery system (DLS liposomes) with episomally replicative DNA plasmids to effect long-term expression of a transgene in cells. A single i.v. injection of a plasmid DNA vector containing the luciferase gene as a marker was administered with the DLS liposomes in BALB/c mice. The luciferase gene and its product were found in all mouse tissues tested as determined by PCR analysis and immunohistochemistry. Luciferase activity was also detected in all tissues tested and was present in lung, liver, spleen, and heart up to 3 months postinjection. In contrast to the nonepisomal vectors tested (pRSV-luc and pCMVintlux), human papovavirus (BKV)-derived episomal vectors showed long-term transgene expression. We found that these episomal vectors replicated extrachromosomally in lung 2 weeks postinjection. Results indicated that transgene expression in specific tissues depended on the promoter element used, DNA/liposome formulation, dose of DNA per injection, and route of administration.

Animals↗

Leukocyte activation in the peripheral blood of patients with cirrhosis of the liver and SIRS. Correlation with serum interleukin-6 levels and organ dysfunction.

OBJECTIVE: Leukocyte adhesion plays an important role in inflammation. Adhesion molecules such as CD11b on polymorphonuclear neutrophil leukocytes (PMNs) up-regulate in response to tumor necrosis factor-alpha, interleukin-8 (IL-8), and other mediators that are involved in systemic inflammatory response syndrome (SIRS). This study examined the behavior of CD11b and other membrane molecules in SIRS in relation to serum cytokines and the severity of illness. DESIGN: Survey study. SETTING: Liver transplantation intensive care unit at a tertiary care center. PATIENTS: A consecutive sample of 22 patients admitted to the liver transplantation intensive care unit for complications related to cirrhosis of the liver in the absence of other disease. Sixteen of the patients developed SIRS and multiple organ dysfunction syndrome with suspected bacterial infections. Seven control subjects were also studied. MAIN OUTCOME MEASURES: Modified Goris organ failure score and Acute Physiology and Chronic Health Evaluation II score. RESULTS: Mean serum IL-6 levels, but not IL-1 beta or tumor necrosis factor-alpha levels, correlated with organ failure (r = 0.79, P < .001). Leukocyte cell-surface markers fluctuated from day to day. The mean of several values was more stable. Mean CD11b and CD35 on PMNs correlated with serum IL-6 level (r = 0.75, P < .001, and r = 0.77, P < .005, respectively). Up-regulation of both CD11b and CD35 display on PMNs correlated with organ failure (r = 0.74, P < .001, and r = 0.71, P < .01, respectively). Polymorphonuclear neutrophil leukocyte L-selectin, CD31, and CD16 were simultaneously decreased, consistent with PMN activation. Monocytes appeared to be activated, but the pattern of surface molecule display was different. CONCLUSIONS: In human SIRS, the circulating monocyte and PMN pools undergo alterations suggestive of leukocyte activation, including up-regulation of PMN CD11b in correlation with the serum IL-6 level and severity of organ dysfunction.

Adult↗

Tumorigenesis and metastasis of neoplastic Kaposi's sarcoma cell line in immunodeficient mice blocked by a human pregnancy hormone.

Kaposi's sarcoma (KS) occurs more often in men than in women and HIV-1-associated KS has a high occurrence in homosexual men (over 30%). Most cultures of KS tumours yield cells with properties of hyperplastic (not malignant) endothelial cells under the control of several cytokines. The role of HIV-1 may be in promoting high levels of some cytokines and providing stimulation to angiogenesis by the HIV-1 Tat protein, which synergizes with basic fibroblast growth factor in promoting these effects. Here we describe an immortalized AIDS-KS cell line (KS Y-1) and show that these cells produce malignant metastatic tumours in nude mice and are killed in vitro and in vivo (apparently by apoptosis) by a pregnancy hormone, the beta-chain of human chorionic gonadotropin. Similarly, chorionic gonadotropin kills KS SLK, cells from another neoplastic cell line (established from a non-HIV-associated KS), as well as the hyperplastic KS cells from clinical specimens grown in short-term culture, but does not kill normal endothelial cells. These results provide evidence that KS can evolve into a malignancy and have implications for the hormonal treatment of this tumour.

Acquired Immunodeficiency Syndrome↗

Use of pulsed-field gel electrophoresis for epidemiological study of Escherichia coli O157:H7 during a food-borne outbreak.

Food and patient isolates from an Escherichia coli O157:H7 outbreak associated with undercooked ground beef were characterized by pulsed-field gel electrophoresis and Shiga-like toxin genotype. Pulsed-field gel electrophoresis confirmed the epidemiologically implicated source of the two-state outbreak and differentiated between outbreak and sporadic strains.

Bacterial Toxins↗

Maternal-fetal interactions affect growth of human immunodeficiency virus type 1 transgenic mice.

Infants vertically infected with human immunodeficiency virus type 1 (HIV-1) often manifest profoundly deficient growth with failure to thrive. The pathologic mechanisms that produce growth failure associated with pediatric HIV infection are not clear. Transgenic mice homozygous for a gag/pol deletion mutant of the infectious provirus pNL4-3 have been found to manifest a similar growth failure pattern. To explore the influence of HIV-1 on fetal growth and maternal-fetal interactions, we examined intrauterine growth of transgenic and nontransgenic mice and evaluated the consequence of embryo transfer into normal and heterozygous transgenic mothers. Mice homozygous for the HIV transgene had normal intrauterine and birth weights but uniformly displayed severe growth retardation postnatally. Transgene expression was prominent in transgenic fetuses and their placentas and in uteri of transgenic mothers, as determined by Northern analysis. Although embryo transfer did not affect intrauterine growth, the pregnancy rate in transgenic mothers was markedly lower than in nontransgenic controls. In both fetal and neonatal tissues, transgene expression was significantly greater in homozygous animals when compared with heterozygotes, but the difference was magnified postnatally. These results suggest that HIV gene expression affected both mother and neonate. In the mother, expression of the HIV-1 transgene reduced postfertilization pregnancy rate. Once the animal was pregnant, however, the effects of transgene expression on the homozygous fetus were overcome in utero, possibly by the contribution of maternal factors or by inhibition of HIV-1 gene expression by a fetal or maternal factor(s). In the neonate, HIV-1 transgene expression increased dramatically in homozygotes and was associated with profound growth failure.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Preventive foot care program: a nursing perspective.

Severe foot problems often develop from minor abnormalities that are preventable. It is crucial for nurses to understand and practice the principles of preventive foot care for elderly patients, both diabetic and non-diabetic. It is also important for nurses to distinguish between lesser foot problems that can easily be treated and more serious conditions that require referral to a specialist. This article includes practical applications for a foot care program from a nursing perspective and discusses pathophysiology of foot injuries, foot assessment using a screening tool and the Semmes-Weinstein monofilaments test, procedural techniques for nail and callus care using a cordless rotary tool, and nurses' qualification for providing nail care.

Diabetic Foot↗

Growth failure and AIDS-like cachexia syndrome in HIV-1 transgenic mice.

The mechanisms which predispose to growth failure in infants and children infected with immunodeficiency virus type-1 (HIV-1) are not fully understood. The contributions of viral replication and CD4+ T cell depletion to growth failure in an HIV-1 transgenic mouse model were investigated. Mice homozygous for the transgene, a gag-pol deletion mutant of the HIV-1 provirus pNL4-3, exhibited marked cachexia, growth retardation, lymphoproliferation with a reduction in the percentage of CD4+ T cells but an increase in the absolute number of splenic CD4+ and CD8+ T cells, thymic hypoplasia, and early death. Despite the absence of T cells, athymic nude mice, homozygous for the HIV transgene, displayed comparable growth failure. The results indicate that AIDS-like cachexia may be produced by expression of viral envelope or accessory genes, need not be accompanied by absolute depletion of CD4+ T cells, and may occur independent of T cell function.

Acquired Immunodeficiency Syndrome↗

bFGF and its low affinity receptors in the pathogenesis of HIV-associated nephropathy in transgenic mice.

HIV-associated nephropathy is characterized by extensive tubulointerstitial disease with epithelial cell injury, microcystic proliferation, and tubular regeneration with glomerulosclerosis. To explore the role of bFGF as a mediator of HIV-induced interstitial disease, we utilized an HIV transgenic mouse model that manifests clinical and histological features observed in patients. In transgenic mice, simultaneous renal epithelial cell proliferation and injury were detected in vivo. In areas of microcystic proliferation, immunoreactive bFGF colocalized with extracellular matrix. Kidneys from transgenic mice had increased bFGF low affinity binding sites, particularly in the renal interstitium. In vitro, transgenic renal tubular epithelial cells proliferated more rapidly and generated tubular structures spontaneously, in marked contrast to nontransgenic renal cells where these pathologic features could be mimicked by exogenous bFGF. These studies suggest that renal bFGF and its receptors play an important role in the pathogenesis of HIV-associated nephropathy.

AIDS-Associated Nephropathy↗

Systemic reactions to immunotherapy: the American Academy of Otolaryngic Allergy morbidity and mortality survey.

Anaphylaxis may be defined as a systemic, immunoglobulin E-mediated (Gell-Coombs type I) hypersensitivity reaction triggered by exposure to an antigen in a previously sensitized patient. Anaphylaxis may occur in a variety of circumstances; however, when it occurs as the result of immunotherapy, it is of great concern to the practicing allergist. When describing or reporting anaphylaxis relating to immunotherapy, most allergists speak in terms of the types of reactions, local vs. systemic. Germane to this discussion is the use of the term systemic reaction, which can mean anything from mild allergy symptoms resulting from an allergy injection to bradycardia and hypotension (shock). In this article we report serious or significant systemic reactions, which are characterized by any of the following symptoms: urticaria, sneezing or nasal obstruction, throat tightness or congestion, wheezing, and shock (bradycardia or hypotension). There were no fatalities reported from the survey group. The overall reaction rate was 0.3%.

Anaphylaxis↗

Children's lunch intake: effects of midmorning snacks varying in energy density and fat content.

To determine the effects of preloads differing in energy density and fat content on subsequent lunch intake, 24 preschool children consumed 113 g ice-cream preloads as mid-morning snacks (containing 0, 12, or 18 g of fat, and 4 g of protein), followed nearly 2 h later by an ad libitum lunch consisting of a variety of foods. A fat-free baseline preload, consisting of dry cereal and apple juice, was also included. The baseline, fat-free, medium and high-fat preloads contained 334, 740, 953, and 1150 kJ per serving. All three ice-cream preloads produced significant but equivalent suppression of ad libitum lunch intake relative to baseline; lunch intake was not related to the fat content or energy density of the ice-cream preloads. Compensation at lunch was not macronutrient specific; children did not increase their fat intake at lunch following the fat-free preload. As energy and fat content of the preloads increased, children consumed significantly fewer foods at lunch, confirming previous findings. These changes in the variety of foods consumed were related to preference: regardless of preload, children continued to consume their preferred foods, while non-preferred foods tended to be eliminated following the higher energy preloads. Because compensation for energy was incomplete, energy intake of meals plus snacks increased with the energy content of the preload. Additional research is required to determine whether manipulating the fat and energy content of snacks produces similar effects when the unit of analysis is the child's total daily diet, rather than a single meal.

Child Nutritional Physiological Phenomena↗

Long-term dietary fish oil supplementation protects against ischemia-reperfusion-induced myocardial dysfunction in isolated rat hearts.

Dietary fish oil has been shown to exert protective effects against arrhythmias and myocardial infarction after coronary artery occlusion. However, the effects of fish oil on ischemia-reperfusion-induced cardiac dysfunction are not known. This study was designed to examine if long-term dietary fish oil protects against a rise in coronary perfusion pressure and myocardial contractile dysfunction following ischemia and reperfusion. Fifteen Sprague-Dawley rats (age 7 to 9 weeks) were fed fish oil-rich chow for 4 to 5 weeks and 11 rats from the same batch were fed ordinary chow. Three fish oil-fed rats were also fed ad libitum indomethacin for the last 2 days. Isolated hearts from both groups were perfused on a Langendorff apparatus and were subjected to 25 minutes of global ischemia and 20 minutes of reperfusion. Myocardial phospholipid acid content was also measured. After 4 to 5 weeks of dietary fish oil supplementation, myocardial content of long-chain polyunsaturated fatty acid (PUFA) (C20-C22) and omega-3 PUFA was increased and that of omega-6 PUFA was decreased in the fish oil-fed group (all p < 0.01). Following global ischemia and reoxygenation, there was a reduction in the force of cardiac contraction and an increase in coronary perfusion pressure. However, reduction in the force of cardiac contraction was less in the hearts of fish oil-fed rats than in the control hearts (49 +/- 9% vs 63 +/- 5%, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cutaneous disorders and viral gene expression in HIV-1 transgenic mice.

Patients infected with HIV-1 experience several hyperproliferative skin disorders, including seborrheic dermatitis, ichthyosis, and psoriasis. Transgenic mice carrying a subgenomic HIV-1 proviral construct lacking the gag and pol genes were found to develop proliferative epidermal lesions, manifested as diffuse epidermal hyperplasia in homozygous transgenic mice and benign papillomas in heterozygous transgenic mice. Nonpapillomatous skin from both homozygotes and heterozygotes expressed viral RNA, and the viral envelope protein gp120 was localized to the suprabasal keratinocyte. Papillomas contained increased amounts of both viral mRNA and envelope glycoprotein. Exposure of transgenic mice to doses of ultraviolet B (UV-B) irradiation that induced cutaneous injury increased viral gene expression and resulted in the development of papillomas within 14-21 days. Cutaneous injury induced by phenol and liquid nitrogen had similar effects. These data support a role for HIV-1 gene products in the pathogenesis of proliferative epidermal disorders associated with HIV-1 infection. Further, they suggest that the process of wound repair increases HIV-1 gene expression in this transgenic mouse model.

Animals↗

Co-purification of 130 kD nitric oxide synthase and a 22 kD link protein from human neutrophils.

The synthesis of nitric oxide (NO) from L-arginine has been demonstrated in several cell types. Both constitutive and inducible forms of NO synthase have been described in different cells. We purified the constitutive form of NO synthase enzyme in human neutrophils using a two-column procedure. Crude 100,000g supernatant of human neutrophils was passed through a 2'-5'-ADP-agarose column followed by a DEAE-Bio-Gel A anion exchange column. NO synthase enzyme migrated as a single band (MW approximately 130,000) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Its activity was dependent upon nicotinamide adenine dinucleotide phosphate (NADPH) and (6R)-tetrahydro-L-biopterin (BH4). In addition, flavin adenine dinucleotide (FAD) was also found to be essential for its maximal activity. A second NADPH, FAD-dependent component (MW approximately 22kD) was also found consistently on the SDS-PAGE gel. These observations suggest co-regulation between NO synthase enzyme and this NADPH, FAD-dependent component, which may be associated with the superoxide radical generating system.

Amino Acid Oxidoreductases↗