Biomedical subjects

R L Simmons

Publications and source records attributed to R L Simmons.

At least 73 records · Page 4Linked to original sources

PubMed1995

Phenotypic analysis of donor cells infiltrating the small intestinal epithelium and spleen during graft-versus-host disease.

One of the principal target organs during graft-versus-host disease (GvHD) is the intestinal epithelium, although the reasons for the preferential involvement of particular organs in this disease are not known. This study analyzed the subset distribution of donor and host lymphocytes in the small intestinal epithelium and the spleen during GvHD in a parent (C57BL/6J) into F1 (C57BL/6JxDBA2/J F1) model. While the donor cell population in the spleen consisted of B and T cells, the donor cell population in the intestine contained only T cells during the course of GvHD. These infiltrating donor cells resembled the host intraepithelial lymphocytes (IELs), which are predominantly CD8+ T cells. This subset distribution of donor cells in the intestinal epithelium was remarkable since they originated from a donor splenocyte population containing few CD8+ lymphocytes. In addition, although the injected donor splenic T cells were virtually all alpha/beta TCR+, several months after GvHD induction more than 30% of the donor cells in the intestine were gamma/delta TCR+, thereby resembling the host IELs not only in their expression of CD4 and CD8, but also in their TCR expression. In contrast, no gamma/delta TCR+ donor cells were detectable in the spleen of GvHD mice. The subset distribution of donor and host IELs remained constant throughout the disease, while in the spleen a decrease of both donor and host B cells and a temporary increase of both donor and host CD8+ cells was observed. These findings demonstrate that in a given target organ during GvHD the disease process affects both donor and host lymphoid populations. In addition the different tissue microenvironments eventually lead to donor cell repopulation with a subset distribution similar to the host natural lymphoid population of the particular target organ.

Animals↗

PubMed1995

The effect of endotoxin on intestinal mucosal permeability to bacteria in vitro.

OBJECTIVE: To examine the role of the intestinal mucosa in bacterial translocation, in vitro bacterial passage across ileal mucosal segments mounted in Ussing chambers were studied in control and endotoxin (lipopolysaccharide)-treated rats. DESIGN: Experimental study. MATERIALS AND METHODS: Three groups of rats were studied. The experimental group received an intraperitoneal injection of lipopolysaccharide, while controls received an equivalent volume of saline solution; a third group received no treatment. Twenty-four hours later, all groups underwent laparotomy and organ culture to assess bacterial translocation. At the same time, a segment of mucosa from the terminal ileum of each animal was mounted in a Ussing chamber, and the transmucosal passage of labeled Escherichia coli from the luminal to serosal surface was assessed by results of serial cultures. RESULTS: In vivo bacterial translocation occurred in 100% of the lipopolysaccharide-treated animals, significantly higher than the incidence seen in controls (25%; P < .05). In vitro passage of labeled E coli across ileal mucosa in the Ussing chamber occurred in 78% of lipopolysaccharide-treated animals, while in controls transmucosal passage was seen in only 14% (P < .05). Histologic examination of mucosa from both groups using light and transmission electron microscopy demonstrated no structural differences between groups. CONCLUSIONS: Increased permeability to bacteria at the mucosal level contributes to the bacterial translocation seen in endotoxemia.

Animals↗

PubMed1995

Cyclic GMP and guanylate cyclase mediate lipopolysaccharide-induced Kupffer cell tumor necrosis factor-alpha synthesis.

Tumor necrosis factor-alpha (TNF-alpha) is an important mediator in sepsis and septic shock. Kupffer cells (KCs) are the resident macrophages of the liver and are potent producers of TNF-alpha in response to inflammatory stimuli such as bacterial endotoxin or lipopolysaccharide (LPS). Although the effects of exogenous cytokines such as interferon-gamma on TNF-alpha production by macrophages have been fairly well studied, the intracellular pathways regulating KC TNF-alpha synthesis are largely unknown. We investigated the role of guanylate cyclase and cGMP in LPS-induced KC TNF-alpha synthesis. Exogenous 8-BrcGMP and dbcGMP increased LPS-stimulated TNF-alpha synthesis but had no effect on KC TNF-alpha in the absence of LPS. Sodium nitroprusside (SNP), a nitric oxide-releasing substance that stimulates guanylate cyclase, increased TNF-alpha synthesis in response to LPS, whereas methylene blue and LY83583, guanylate cyclase inhibitors, decreased KC TNF-alpha synthesis. The inhibitory effect of methylene blue could be overcome with exogenous dbcGMP or SNP. Our results demonstrate that guanylate cyclase and cGMP mediate LPS-induced KC TNF-alpha synthesis and suggest that agents that alter cyclic nucleotide metabolism in KCs may affect the response of these cells to inflammation and inflammatory stimuli.

Aminoquinolines↗

PubMed1995

Prostanoids inhibit Kupffer cell nitric oxide synthesis.

Kupffer cells are the largest population of fixed tissue macrophages in the body and produce a number of mediators that are involved in host defense. These mediators include cytokines such as tumor necrosis factor-alpha and interleukin-1, prostaglandins, oxygen radicals, and nitric oxide. Prostaglandins are produced by adjacent endothelial cells in addition to Kupffer cells and regulate a number of cellular functions in a wide array of cells, but their role in nitric oxide synthesis is controversial. We studied the role of prostaglandins in regulating lipopolysaccharide (LPS)-induced nitric oxide synthesis in cultured rat Kupffer cells. Prostaglandin E2 (PGE2) inhibited Kupffer cell nitric oxide synthesis in a dose-dependent fashion in both 24- and 48-hr cultures. The effect of PGE2 persisted at low and high LPS concentrations. Prostaglandin analogues as well as other prostanoids also inhibited Kupffer cell nitric oxide synthesis. These data show that exogenous prostaglandins suppress Kupffer cell nitric oxide synthesis and may represent an important endogenous regulator of nitric oxide production.

Animals↗

PubMed1995

The effect of nitric oxide on fetal pulmonary artery smooth muscle growth.

Endothelium-derived nitric oxide (NO) relaxes fetal pulmonary arterial vessels through activation of guanylate cyclase and increasing smooth muscle cyclic guanosine 3', 5'-monophosphate (cGMP). Exogenous NO administered as a gas at low concentrations shares this effect, decreasing pulmonary artery resistance and increasing in pulmonary blood flow. NO, endogenously synthesized or inhaled as a gas, may affect cellular growth in the underlying pulmonary vascular smooth muscle media. We report the effects of NO and cGMP upon DNA synthesis and proliferation of passaged pulmonary vascular smooth muscle cells from fetal rats. Smooth muscle cells from rat fetal pulmonary artery (RFPASM; 18-19 day gestation; term 21 days) were treated in culture with sodium nitroprusside (SNP), isosorbide dinitrite (ISDN)--both NO-generating vasodilators--or 8-bromo-cGMP, a cell-permeant cGMP analog. All agents inhibited thymidine uptake at concentrations of 10(-3)-10(-2) M. Lower concentrations (10(-5)-10(-4) M) of SNP and ISDN increased [3H]-thymidine ([3H]TdR) uptake, an effect not seen with cGMP at similar concentrations. Exposing RFPASM to authentic NO gas in a deoxygenated medium inhibited [3H]TdR uptake only. NO appears to have a biphasic effect on DNA synthesis in passaged RFPASM, with stimulation at micromolar concentrations and inhibition at higher levels. NO may thus alter vascular smooth muscle growth and pulmonary vascular remodeling in conditions complicated by pulmonary hypertension and treated with inhaled NO.

Animals↗

PubMed1995

Characterization of inducible nitric oxide synthase expression in endotoxemic rat cardiac myocytes in vivo and following cytokine exposure in vitro.

Lipopolysaccharide (LPS) treatment results in widespread expression of the inducible isoform of nitric oxide (NO) synthase (iNOS). Although there is evidence for the expression of iNOS in heart tissue, regulation of myocardial iNOS expression is not known. To determine the time course and degree of iNOS induction in the adult heart, we examined iNOS mRNA expression and enzyme activity in (1) rat left ventricular tissue after LPS treatment in vivo, and (2) cultured, long-term rat cardiac myocytes maintained in serum and exposed to interleukin-1 beta, tumor necrosis factor-alpha, interferon-gamma, and/or LPS. iNOS mRNA was detected by Northern blot analysis and in situ hybridization. iNOS enzyme activity was measured in extracts of whole heart, and nitrate and nitrite (the stable end-products of NO) accumulation was quantified in cardiomyocyte culture media. iNOS mRNA was not detected in untreated hearts or cultured myocytes but was apparent within 3 h in both hearts obtained from LPS-treated animals and in cytokine-treated myocytes. In whole heart, iNOS mRNA expression peaked by 6 h after LPS and declined by 12 and 24 h. In situ hybridization demonstrated perinuclear localization of iNOS mRNA in both cardiac vascular smooth muscle and myocytes with maximal expression at 6 h after LPS injection. In cardiac myocytes, iNOS expression was maximal at 12 to 24 h, persisted through 48 h, and was partially inhibited by dexamethasone. Interferon-gamma was the most potent single cytokine with regards to myocyte iNOS induction. Nitric oxide release in cytokine-stimulated cardiac myocytes was largely in the form of nitrate and was associated with increased glucose uptake and lactate release; the former finding indicates that NO interacts with myocardial heme proteins and/or oxyradicals, while the latter suggests inhibition of oxidative metabolism. Although non-myocardial cells may significantly contribute to iNOS expression in whole heart tissue, significant iNOS expression and NO production also take place within the myocyte. Induced NO production may regulate myocardial perfusion and impair myocardial function and metabolism.

Animals↗

PubMed1995

Hemorrhagic shock.

A great deal has been learned about the pathophysiologic condition of hemorrhagic shock. The response of the hormonal and inflammatory mediator systems in patients in hemorrhagic shock appears to represent a distinct set of responses different from those of other forms of shock. The classic neuroendocrine response to hemorrhage attempts to maintain perfusion to the heart and brain, often at the expense of other organ systems. This intense vasoconstriction occurs via central mechanisms. The response of the peripheral microcirculation is driven by local tissue hypoperfusion that results in vasodilation in the ischemic tissue bed. Activation of the systemic inflammatory response by hemorrhage and tissue injury is an important component of the pathophysiologic condition of hemorrhagic shock. Activators of this systemic inflammatory response include ischemia/reperfusion injury and neutrophil activation. Capillary "no-flow" with prolonged ischemia and "no-reflow" with reperfusion may initiate neutrophil activation in patients in hemorrhagic shock. The mechanisms that lead to decompensated and irreversible hemorrhagic shock include (1) "arteriolar hyposensitivity" as manifested by progressive arteriolar vasodilation and decreased responsiveness of the microcirculation to alpha-agonists, and (2) cellular injury and activation of both proinflammatory and counterinflammatory mechanisms. These changes represent a failure of the microcirculation. Redistribution of cardiac output and persistent gut ischemia after adequate resuscitation may also contribute to the development of irreversible hemorrhagic shock. Treatment of hemorrhagic shock includes rapid operative resuscitation to limit activation of the mediator systems and abort the microcirculatory changes that result from hemorrhagic shock. Volume resuscitation and control of hemorrhage, should occur simultaneously. The end point in volume resuscitation of hemorrhagic shock must be maintenance of organ system and cellular function. Whether we use adequate urine output, correction of lactic acidemia, optimization of oxygen delivery, or oxygen consumption as our specific goal, the general objective is to provide adequate crystalloid solution and packed red blood cells to achieve and maintain normal organ and cellular perfusion and function.

Adrenal Glands↗

PubMed1995

Use of visual perception in estimating static postural stresses: magnitudes and sources of errors.

Very little is known about the magnitudes and sources of errors associated with the visual estimation of postural classification displayed on TV screens. This study was conducted to address this issue. Sixty-three subjects participated in the experiments. The findings indicate that: (1) subjects found it difficult to evaluate upper extremity postures (particularly the elbow and the wrist), while the postures around the lower back were the easiest to evaluate; (2) the lower extremity positions affected the ability of the subjects to accurately classify postures around the wrist, elbow, shoulder, neck, and lower back, with the estimates being > 70% for sitting and > 60% for standing (except for the elbow); and (3) in general, flexion and extension are easier to evaluate than neutral and non-neutral postures.

Adult↗

PubMed1995

cAMP enhances inducible nitric oxide synthase mRNA stability in cardiac myocytes.

The effects of adenosine 3',5'-cyclic monophosphate (cAMP) on cardiac myocyte nitric oxide (NO) production were studied. Maximal nitrite (NO2(-)) production by cultured neonatal rat cardiac myocytes was achieved with 500 U/ml interleukin-1 beta (IL-1 beta) for 48 h (4.6 +/- 0.3 nmol/1.25 x 10(5) cells; n = 12). Cardiac myocytes exposed to 500 U/ml IL-1 beta for 48 h stained positively for inducible nitric oxide synthase (iNOS) by immunohistochemistry. Forskolin (FSK; adenylate cyclase stimulator) or dibutyryl cAMP (DBcAMP; membrane-permeable cAMP analogue) administration alone had no effect on NO2(-) production. The addition of FSK or DBcAMP to IL-1 beta significantly increased NO2-) levels vs. IL-1 beta alone (9.7 +/- 0.6 and 10.9 +/- 0.8 vs. 4.6 +/- 0.3 nmol/1.25 x 10(5) cells per 48 h, respectively; P < 0.01; n = 12). Semiquantitative reverse transcriptase-polymerase chain reaction revealed increased iNOS mRNA in myocytes treated with FSK+IL-1 beta or DBcAMP+IL-1 beta vs. those treated with IL-1 beta alone. The addition of FSK or DBcAMP to IL-1 beta increased iNOS mRNA half-life over IL-1 beta treatment alone (10.6, 11.7 vs. 2.4 h, respectively). Cardiac myocytes do not express iNOS in response to cAMP alone. Rather, cAMP enhances iNOS mRNA stability following cytokine exposure.

Animals↗

PubMed1995

Lipopolysaccharide binding protein participation in cellular activation by LPS.

Lipopolysaccharide binding protein (LBP) is a plasma protein that plays an important intermediary role in host-endotoxin (LPS) interactions. LBP binds with high affinity to the lipid A portion of LPS and then interacts with the monocytic differentiation antigen CD14 to markedly up-regulate TNF-alpha production by mononuclear phagocytes. In the presence of LBP, 100-fold less LPS is required to trigger this cytokine response. LBP has been implicated in the interaction of LPS with both CD14+ (monocytes, macrophages, neutrophils) and CD14-cells (endothelial and epithelial cells) to promote such varying responses as secretion of cytokines and nitric oxide (NO), expression of adhesion molecules, production of tissue factor, and activation of neutrophils. Non-CD 14-bearing cells, such as endothelial cells, can also respond to LPS through this pathway via the soluble form of CD14, an interaction that is similarly enhanced by LBP. Recently, it has been proposed that LBP cannot only potentiate host responses to LPS but can also facilitate the neutralization of LPS under certain conditions. LBP has been described as acting as a lipoprotein transfer protein facilitating the transfer of LPS both to the target receptor (CD14) and lipoprotein (HDL).

Acute-Phase Proteins↗

PubMed1995

Nitric oxide synthase inhibitor alters papillary muscle force-frequency relationship.

We provide evidence for an immediate effect of NG-monomethyl-L-arginine (L-NMMA) on the force-frequency relationship in isolated hamster papillary muscles. L-NMMA (competitive inhibitor of nitric oxide synthase) reversed the force-frequency relationship (staircase effect) in isolated hamster papillary muscles from negative to positive (P < .01; ANOVA; n = 6). The addition of L-arginine (substrate for nitric oxide synthase) blocked the L-NMMA effect (P < .01; ANOVA; n = 6). The addition of the nitric oxide (NO) donor, sodium nitroprusside (NTP), significantly increased the level of cGMP in the tissue bath (P < .01; test; n = 6) and reversed the positive inotropic effect of L-NMMA on staircase (P < .01; ANOVA; n = 6). The addition of 8-Br-cGMP to the bath resulted in a concentration-dependent decrease in tension generated by the papillary muscles (n = 6). Methylene blue (known inhibitor of cGMP) mimicked the effect of L-NMMA on staircase (P < .01; ANOVA; n = 6). L-NMMA also significantly blunted the negative inotropic effect of ryanodine (SR calcium release channel regulator) (P < .01; ANOVA; n = 6). The positive inotropic effect of Bay K 8644 (sarcolemmal, L-type calcium channel regulator) was not affected by L-NMMA (P = NS; ANOVA; n = 6). L-NMMA had no effect on either [3H]ryanodine or [3H]PN200-110 (sarcolemmal, L-type calcium channel regulator) binding to cardiac membranes. These findings support a cGMP-dependent role for endogenous NO in myocardial E-C coupling.

Amino Acid Oxidoreductases↗

PubMed1994

Chronotropic effects of cytokines and the nitric oxide synthase inhibitor, L-NMMA, on cardiac myocytes.

We and others have proposed that cytokine-stimulated nitric oxide (NO) production is responsible for reversible myocardial depression in sepsis, trauma and ischemia. An effect of NO on cardiac sarcolemmal L-type calcium channels has also recently been proposed. The spontaneous beating rate of neonatal cardiac myocytes is regulated by the sarcolemmal L-type calcium channel. Accordingly, we sought to determine if cytokine-stimulated NO production could also regulate beating rates of neonatal cardiac myocytes. Treatment of neonatal rat cardiac myocytes with TNF, IL-1, IL-6, 10(-5)M NMA, or 10(-3)M NMA significantly enhanced spontaneous beating rates compared to untreated myocytes in serum-free media for 48 hours (p < or = .01; n = 12 for each). Only IL-1 treatment resulted in significant nitrite levels vs. control over 48 hours (4.2 +/- 0.7 vs. 0.3 +/- 0.2 nmoles/1.25 x 10(-5) cells, respectively) (n = 12). Nitrite production by IL-1 was inhibited by 10(-3)M NMA but not 10(-5)M NMA (0.3 +/- 0.2 vs. 4.1 +/- 0.6 nmoles; p < .01; n = 12). The addition of 10(-5)M NMA to TNF, IL-1, and IL-6 did not alter the effect of the cytokines on the spontaneous beating rates of the cardiac cells (p < or = .01; n = 12 for each). These results strongly suggest that cytokines and NMA affect cardiac myocyte spontaneous beating rates through mechanisms independent of NO.

Amino Acid Oxidoreductases↗

PubMed1994

Phenotypic characterization of a novel bone marrow-derived cell that facilitates engraftment of allogeneic bone marrow stem cells.

Bone marrow transplantation is an accepted therapy for hematologic malignancies, aplastic anemia, metabolic disorders, and solid tumors. However, graft-versus-host disease (GVHD) and failure of engraftment have limited the widespread application of this technology to nonmalignant disease states. The use of purified bone marrow stem cells has been suggested as an approach to promote engraftment yet avoid GVHD. Although bone marrow stem cells, purified by cell sorting, engraft and repopulate lethally irradiated genetically identical recipients, they do not engraft in major histocompatibility complex (MHC)-disparate allogeneic recipients. We report for the first time the characterization of a novel cell population of donor bone marrow origin, separate from the hematopoietic stem cell, that facilitates engraftment of purified allogeneic bone marrow stem cells in an MHC-specific fashion without causing GVHD. Although 1,000 purified stem cells (c-kit+/Sca-1+/lineage-) reliably repopulate syngeneic mouse recipients, 10 times that number do not engraft in MHC-disparate allogeneic recipients. The addition of as few as 30,000 facilitating cells (CD8+/CD45R+/TCR-) is sufficient to permit engraftment of purified stem cells in MHC-disparate recipients. The cell surface phenotype of this purified cellular population differs significantly from other characterized lineages of lymphoid or myeloid origin. Based on multiparameter rare-events cell sorting, the facilitating fraction is CD8+, CD3+, CD45R+, Thy 1+, class IIdim/intermediate but alpha beta-TCR- and gamma delta-TCR-. This cellular population comprises approximately 0.4% of the total bone marrow and is separate from the hematopoietic stem cell. The coadministration of purified facilitating cells plus stem cells to optimize engraftment yet avoid GVHD may expand the potential application of bone marrow transplantation to disease states in which the morbidity and mortality associated with conventional BMT cannot be justified.

Animals↗

PubMed1994

Catalytic conversion of L-arginine to nitrate by platinum electrodes.

Platinum electrodes (PLE) have been used recently to study the physiologic effects of the enzymatic conversion of L-arginine (L-arg) to nitric oxide (NO) by nitric oxide synthase (NOS). We sought to determine if PLE alone could catalytically convert L-arg to NO without NOS. Accordingly, NO2- + NO3- levels (NOx) were measured by HPLC from a physiologic Tyrode's buffer in the presence and absence of electrical stimulation with PLE or silver chloride electrodes (SCE) for 10 min (n = 6 for each condition). PLE stimulated at 1Hz produced 24 +/- 0.7 and 15.4 +/- 2.8 microM NOx from D-arg and L-arg (100mM) respectively. PLE + L-arg stimulated from 0 to 5 Hz produced increasing NOx (2.9 +/- 0.27;15.4 +/- 2.8;19.7 +/- 4;32 +/- 5;37 +/- 6.2;43 +/- 7 microM). SCE+L-arg showed no increase in NOx above background. The addition of 1% BSA to the Tyrode's buffer further increased NOx levels. NO2- levels were undetectable, indicating all NOx levels were NO3-. We conclude that PLE can catalytically convert L-Arg to NO3- via electron transfer without NOS.

Arginine↗

PubMed1994

Molecular cloning, characterization, and tissue distribution of rat lipopolysaccharide binding protein. Evidence for extrahepatic expression.

LPS binding protein (LBP) is a glycoprotein present in normal serum that becomes markedly elevated during acute phase responses. LBP has been reported to greatly potentiate host responses to endotoxin or LPS. Therefore, LBP may play a critical role in the body's response to injury and infection. Little is known about the factors regulating production of LBP. To investigate the regulation of LBP expression, we have cloned the full-length cDNA for rat LBP. The deduced amino acid sequence of rat LBP was highly homologous with that reported for rabbit and human LBP. The sequence of rat LBP further refines the conserved regions found within the family of proteins that bind LPS; this family is comprised of bactericidal permeability-increasing protein and LBP from multiple species. Use of the rat LBP cDNA clone for Northern blot analysis reveals that LBP mRNA levels are markedly up-regulated in liver during acute phase responses. However, in contrast to previous reports, we also find evidence of extrahepatic expression of LBP under these induced conditions. The presence of LBP mRNA in activated tissues other than liver suggests that LBP may play a larger role in local tissue responses to LPS than previously appreciated.

Acute-Phase Proteins↗

PubMed1994

Screening donors for xenotransplantation. The potential for xenozoonoses.

Xenotransplantation is a potential solution to the current donor shortage for solid organ transplantation. The transmission of infectious agents from donor organs or bone marrow to the recipient is a well-recognized phenomenon following allotransplantation. Thus the prospect of xenotransplantation raises the issue of xenozoonoses--i.e., the transmission of animal infections to the human host. Anticipating an increasing number of baboon to human transplants, 31 adult male baboons (Papio cynocephalus) from a single colony in the United States were screened for the presence of antibody to microbial agents (principally viral) that may pose a significant risk of infection. Antibody to simian cytomegalovirus, simian agent 8 and Epstein-Barr virus, was found in 97% of animals tested. Antibody to simian retroviruses and Toxoplasma gondii was found in 30% and 32% respectively. Discordant results were found when paired samples were examined by two primate laboratories. This was particularly noted when methodologies were based on cross-reaction with human viral antigens. These results highlight the need to develop specific antibody tests against the species used for xenotransplantation.

Animals↗

PubMed1994

Differential induction of nitric oxide synthase in hepatocytes during endotoxemia and the acute-phase response.

OBJECTIVE: Nitric oxide (NO) is a potent biologic mediator produced by hepatocytes following exposure to cytokines and lipopolysaccharide (LPS). These cytokines are also known to regulate induction of the hepatic acute-phase response. The objective of this study was to determine whether inducible nitric oxide synthase (iNOS), the enzyme that produces NO, is expressed as part of the hepatic acute-phase response. DESIGN: The gene expression for inducible NOS (iNOS) as well as alpha 1-acid glycoprotein (AGP), an established acute-phase reactant, was measured by Northern blot analysis in rat hepatocytes in vivo during endotoxemia (LPS injection) and during the acute-phase response produced by hindlimb turpentine injection. Hepatocyte iNOS messenger RNA (mRNA) levels were correlated with iNOS activity and circulating plasma nitrite and nitrate levels. In vitro, iNOS and AGP mRNA levels were determined in cultured hepatocytes stimulated with interleukin 6 (IL-6), interleukin 1 beta (IL-1 beta), tumor necrosis factor alpha (TNF-alpha), or dexamethasone. RESULTS: The AGP mRNA levels were increased in vivo following both LPS and turpentine injection, while iNOS expression was induced only by LPS injection. Hepatocyte iNOS activity and plasma nitrite and nitrate levels also increased after LPS treatment. In vitro, the cytokine combination IL-6, IL-1 beta, and TNF-alpha induced hepatocyte iNOS expression but had minimal effects on AGP in the absence of dexamethasone. Addition of dexamethasone alone markedly increased AGP mRNA levels, with further increases seen with TNF-alpha or IL-1 beta addition. In contrast, dexamethasone decreased iNOS expression. CONCLUSION: The results show that hepatocyte iNOS expression is not part of the acute-phase response induced by remote inflammation and indicates that iNOS is differentially regulated from the acute-phase reactant, AGP.

Acute-Phase Reaction↗

PubMed1994

Quantitative and morphologic analysis of bacterial translocation in neonates.

OBJECTIVE: To elucidate the mechanisms of bacterial translocation in animals fed a conventional formula by correlating transmucosal bacterial passage in vitro with the structural characteristics of the neonatal intestinal mucosa. DESIGN: Newborn rabbits were randomized to receive a conventional formula or breast milk. Bacterial translocation to the mesenteric lymph nodes, liver, and spleen was quantitated after 7 days, and transmucosal passage of bacteria was measured in vitro using the Ussing chamber. The mucosal membranes were examined by light, transmission electron, and confocal laser scanning microscopy. RESULTS: Bacterial passage was rarely seen in the breast milk-fed animals in contrast to the formula-fed animals. Unlike the normal-appearing membranes from breast milk-fed animals, the epithelial cells of formula-fed animals were vacuolated but healthy, with normal polarization and microvillus border by confocal laser scanning microscopy. Villi of formula-fed animals were less densely packed than those of the breast milk-fed animals. Bacterial adhesion, internalization, and transmucosal passage were seen only in membranes from formula-fed animals. Transmission electron microscopy demonstrated bacteria incorporating into the epithelial surface through an active phagocytic process, with rearrangement of the actin cytoskeleton. Once internalized, these bacteria were seen within the cytoplasmic vacuoles and subsequently in the submucosa. No bacteria passed between epithelial cells. CONCLUSION: Morphological changes in the intestinal mucosa of formula-fed newborn rabbits may increase permeability to bacteria.

Animals↗