RESUMEN
Haemolysis of serially collected insulin serum samples frequently causes falsely-low measured concentrations because of the release of intracellular insulin degrading enzyme (IDE). We investigated if bacitracin, an in vitro IDE inhibitor, could prevent haemolysis-induced insulin degradation during insulin sensitivity testing. Blood samples were collected from adults undergoing serial sampling for insulin sensitivity. A dose-finding study measured insulin from experimentally haemolysed samples containing five bacitracin concentrations (0-2.5 g/L) and from non-experimentally haemolysed samples. To confirm the utility of bacitracin in the clinical setting, we compared insulin in samples collected with and without 1 g/L bacitracin from a frequently sampled intravenous glucose tolerance test (FSIVGTT), where haemolysis often occurs accidentally. In the dose-finding study, bacitracin 0.25, 1 and 2.5 g/L all maximally prevented insulin degradation in experimentally haemolysed samples. Among FSIVGTT unintentionally haemolysed samples, insulin concentrations from bacitracin-containing samples were significantly higher than from those without bacitracin (P < .01), and not different from non-haemolysed samples obtained simultaneously from a second intravenous catheter (P = .07). Bacitracin did not significantly alter insulin concentrations in non-haemolysed samples. Bacitracin attenuates haemolysis-associated insulin degradation in clinical samples, enabling a more accurate assessment of insulin sensitivity and glucose homeostasis.
Asunto(s)
Resistencia a la Insulina , Preparaciones Farmacéuticas , Adulto , Bacitracina , Reposicionamiento de Medicamentos , Hemólisis , Humanos , InsulinaRESUMEN
Lack of enteral feeding increases P- and E-selectin and ICAM-1 expression on endothelial cells in organs, such as the small intestine and lung, and increases neutrophils in the intestine. These changes are associated with increased mortality after gut ischemia. We hypothesize that nutritional regimen affects endothelial ICAM-1 levels and leukocyte beta2 integrins after gut ischemia. Mice received chow, intravenous (IV) TPN, or intragastric (IG) TPN. In experiment 1, after 5 days of diet, 28 mice underwent 15 min of superior mesenteric artery (SMA) occlusion (I/R) for quantification of ICAM-1 expression in organs 3 h later. In experiment 2, after the same nutrient pretreatments of 38 mice, peripheral blood was obtained with or without gut I/R to measure CD11a and CD11b expression on myeloid cells. CD18 immunofluorescence staining was studied in the lung. Expression of ICAM-1 in the liver, kidney, and small intestine was significantly higher after IV-TPN than chow. IG-TPN reduced liver and kidney ICAM-1 levels midway between the chow and IV-TPN groups, but not intestinal expression. Expression of CD11b on the myeloid cell population in each group was similar before I/R, but CD11b levels increased after IV-TPN on circulating cells after I/R compared with all uninjured animals or injured chow or IG-TPN mice. Only IV-TPN mice had lung CD18-positive leukocytes after I/R. After I/R, lack of enteral feeding increases organ expression of ICAM-1, CD11b levels on myeloid cells, and lung of CD18 positive leukocytes. Through these changes, lack of enteral feeding may increase organ damage after gut ischemia.
Asunto(s)
Cadenas beta de Integrinas/metabolismo , Molécula 1 de Adhesión Intercelular/metabolismo , Intestino Delgado/irrigación sanguínea , Isquemia/diagnóstico , Insuficiencia Multiorgánica/prevención & control , Nutrición Parenteral , Análisis de Varianza , Animales , Biomarcadores/análisis , Antígenos CD11/análisis , Antígenos CD18/análisis , Modelos Animales de Enfermedad , Citometría de Flujo , Cadenas beta de Integrinas/análisis , Molécula 1 de Adhesión Intercelular/análisis , Mucosa Intestinal/patología , Intestino Delgado/patología , Masculino , Ratones , Ratones Endogámicos ICR , Probabilidad , Pronóstico , Valores de Referencia , Sensibilidad y EspecificidadRESUMEN
Clinical and laboratory evidence shows that enteral feeding significantly reduces pneumonia and intra-abdominal abscess formation after celiotomy for severe trauma. Supplementation of total parenteral nutrition (TPN) with glutamine (GLN) supports impaired immunity induced by TPN in several animal and human studies. This work investigates the peritoneal cellular response and polymorphonuclear neutrophil (PMN) bactericidal function after mouse chemical peritonitis after TPN with and without GLN. Thirty-three mice received chow, TPN, or 2% GLN-supplemented TPN (GLN-TPN) for 5 days. All mice then received 2 mL of a 1% glycogen solution intraperitoneally to induce cell exudation, and peritoneal exudative cells (PECs) were recovered 4 h later. Total and differential PEC numbers, as well as PMN phagocytosis, reactive oxygen intermediate production (ROI), CD11b (integrin aM chain) expression, and CD16/32 (Fcgamma II/III receptor) expression were measured. PMN, macrophage, and lymphocyte cell numbers were significantly lower with TPN than with chow or GLN-TPN groups, with no differences between chow and GLN-TPN. TPN significantly lowered peritoneal PMN phagocytosis compared with chow (P < 0.05) and approached significance with GLN-TPN (P = 0.06). There were no significant differences in ROI production or CD11b and CD16/32 expression on peritoneal PMN. GLN supplementation improved the reduction in cell exudation and PMN phagocytosis induced by TPN after chemical peritonitis.
Asunto(s)
Glutatión/farmacología , Glucógeno/farmacología , Neutrófilos/metabolismo , Neutrófilos/patología , Animales , Peso Corporal , Antígeno CD11b/biosíntesis , Adhesión Celular , Citometría de Flujo , Glutatión/metabolismo , Glucógeno/metabolismo , Inflamación , Macrófagos/metabolismo , Ratones , Fagocitosis , Especies Reactivas de Oxígeno , Receptores de IgG/biosíntesisRESUMEN
BACKGROUND: The Th2 cytokines, interleukin-4 (IL-4) and interleukin-10 (IL-10), stimulate IgA production. Total parenteral nutrition (TPN) reduces IL-4 and IL-10 messenger RNA in gut lamina propria lymphocytes, total IL-4 and IL-10 in gut homogenates, and IgA-mediated mucosal immunity. Bombesin (BBS) can maintain mucosal immunity in TPN-fed mice, but the effects of BBS on gut IL-4 and IL-10 levels and their mRNA expression in the lamina propria are unknown. METHODS: In experiment 1, mice that were fed chow, TPN, or TPN + BBS (15 microg/kg intravenously-three times a day) for 5 days were killed, and respiratory tract IgA and intestinal IgA, IL-4, and IL-10 levels were measured. In experiment 2, IL-4 and IL-10 mRNA were measured in isolated lamina propria lymphocytes from chow-, TPN-, and TPN+BBS-fed mice by reverse transcriptase-polymerase chain reaction. Intestines were harvested 1 hour after injection of 100 7 microg of lipopolysaccharide intraperitoneally. Samples were standardized to beta-actin. RESULTS: TPN-fed mice had significantly lower respiratory tract IgA levels than chow- or TPN + BBS-fed mice. TPN+BBS did not increase intestinal IL-10 or IL-10 lamina propria mRNA levels but maintained intestinal IL-4 levels and lamina propria IL-4 mRNA expression equal to those of chow-fed mice. CONCLUSIONS: BBS reverses the effects of TPN on intestinal and respiratory tract IgA levels and most effects on gut cytokines. Lamina propria cytokine mRNA levels reflect total gut cytokine concentration.
Asunto(s)
Bombesina/farmacología , Inmunoglobulina A Secretora/inmunología , Interleucina-4/análisis , Mucosa Intestinal/inmunología , Nutrición Parenteral Total , Animales , Inmunidad Mucosa , Interleucina-10/genética , Interleucina-4/genética , Masculino , Ratones , ARN Mensajero/análisisRESUMEN
BACKGROUND: Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is an adhesion molecule that directs naive T and B cells into Peyer's patches for sensitization and distribution to intestinal and extraintestinal sites. With no enteral stimulation, its expression drops rapidly in association with reduced Peyer's patch cell populations and increases rapidly with reinstitution of enteral feeding. Because both glutamine (GLN) and bombesin (BBS) preserve mucosal immunity, this study examined whether they preserve MAdCAM-1 expression. METHODS: In 2 separate experiments, animals were randomized to IV cannulation with chow, total parenteral nutrition (TPN), and (experiment 1) 15 microg/kg BBS 3 times per day or (experiment 2) an isocaloric, isonitrogenous 2% GLN-supplemented solution. After 5 days of feeding, MAdCAM-1 expression in Peyer's patches, spleen, and intestine was measured using a dual radiolabeled monoclonal antibody technique. RESULTS: MAdCAM-1 expression was not significantly improved from TPN levels either with BBS or GLN supplementation. Levels of MAdCAM-1 expression remained unchanged in non-Peyer's patch sites. CONCLUSIONS: Although MAdCAM-1 is considered the gateway molecule for cell entry into mucosal immunity, this does not seem to be the mechanism for mucosal immunity preservation in nonenterally fed mice receiving bombesin or glutamine.
Asunto(s)
Bombesina/farmacología , Glutamina/farmacología , Inmunidad Mucosa/efectos de los fármacos , Inmunoglobulinas/metabolismo , Mucoproteínas/metabolismo , Nutrición Parenteral Total , Animales , Moléculas de Adhesión Celular , Inmunoglobulinas/efectos de los fármacos , Intestino Delgado/efectos de los fármacos , Intestino Delgado/inmunología , Intestino Delgado/metabolismo , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/metabolismo , Masculino , Mesenterio , Ratones , Ratones Endogámicos ICR , Mucoproteínas/efectos de los fármacos , Nutrición Parenteral Total/efectos adversos , Ganglios Linfáticos Agregados/efectos de los fármacos , Ganglios Linfáticos Agregados/inmunología , Ganglios Linfáticos Agregados/metabolismo , Distribución Aleatoria , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/metabolismoRESUMEN
BACKGROUND: Total parenteral nutrition (TPN) alters gut cytokines and mucosal immunity and increases intercellular adhesion molecule-1 (ICAM-1) expression, gut neutrophil levels, and mortality after gut ischemia. Supplementation of TPN with glutamine partially supports mucosal immunity by preserving respiratory and intestinal IgA levels, maintaining the proper IgA-stimulating cytokine milieu within the intestine, and reducing intestinal ICAM-1 expression and neutrophil accumulation. This work investigates whether glutamine supplementation of TPN affects mortality in mice after gut ischemic insult. METHODS: Thirty-eight mice were randomized to receive chow, TPN, or 2% glutamine-supplemented TPN (GLN-TPN) for 5 days. After feeding their respective diets, gut ischemia/reperfusion (I/R) was induced with superior mesenteric artery occlusion for 30 minutes followed by resuscitation with 1 mL saline. Survival was recorded until 72 hours after reperfusion. RESULTS: Survival time was significantly reduced in the TPN-fed mice compared with both chow-fed and GLN-TPN-fed mice (p < .05). Survival at 72 hours after reperfusion was also significantly lower in the TPN-fed mice than in the chow-fed and GLN-TPN-fed mice (p < .05) CONCLUSIONS: Glutamine supplementation of TPN significantly improves survival after gut I/R, suggesting modulation of the inflammatory response or improved gut tolerance to low-flow states.
Asunto(s)
Glutamina/administración & dosificación , Molécula 1 de Adhesión Intercelular/metabolismo , Nutrición Parenteral Total , Daño por Reperfusión/terapia , Animales , Citocinas/análisis , Glutamina/farmacología , Inmunidad Mucosa/efectos de los fármacos , Inmunoglobulina A/inmunología , Molécula 1 de Adhesión Intercelular/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos ICR , Distribución Aleatoria , Análisis de SupervivenciaRESUMEN
Single and double dichotic digit tests in Malay language were developed and standardized as an initial attempt to incorporate tests of auditory processing within the scope of audiology practice in Malaysia. Normative data under free recall, directed right-ear first, and directed left-ear first listening conditions were determined using 120 Malay children between the ages of 6 and 11 years old with normal hearing and normal academic performance. Test-retest reliability was assessed in 15 of the study subjects. In general, the double dichotic digit test produced greater differences in scores between age groups, and a greater right-ear advantage than the single dichotic digit test. In addition, the double dichotic digit test also had higher test-retest reliability. These findings suggest the double dichotic digit test is more clinically applicable.
Asunto(s)
Trastornos de la Percepción Auditiva/diagnóstico , Pruebas de Audición Dicótica/normas , Lenguaje , Estimulación Acústica , Niño , Pruebas de Audición Dicótica/métodos , Femenino , Humanos , Malasia , Masculino , Desarrollo de Programa , Estándares de Referencia , Valores de Referencia , Reproducibilidad de los ResultadosRESUMEN
OBJECTIVE: To assess whether lack of enteral feeding significantly impairs generation of specific immune responses to an acute viral infection. SUMMARY BACKGROUND DATA: Parenteral feeding provides adequate nutrients to meet metabolic needs, but lack of enteral stimulation creates a defect in mucosal immunity characterized by loss of IgA-mediated defenses in the respiratory tract. METHODS: The enzyme-linked Immunospot (ELISPOT) assay was used to determine accumulation of immunologic cells in the nasal passages after diet manipulation. Viral shedding and nasal IgA levels were measured in additional groups of mice. RESULTS: After determining the time course of antibody-forming cells (AFCs) via ELISPOT to an active infection with the A/PR8 influenza virus, a significant reduction was found in total AFCs, IgA-producing AFCs, and IgG-producing AFCs over the course of a 13-day experiment with significant depression in viral-specific respiratory IgA levels. Eight days following an active infection, seven of nine total parenteral nutrition-fed animals continued to have viral shedding in the nasal passages compared to one of nine chow-fed mice and one of six animals fed a complex enteral diet. CONCLUSIONS: Lack of enteral stimulation significantly impairs the generation of IgA-mediated mucosal immunity.
Asunto(s)
Formación de Anticuerpos , Alimentos , Nutrición Parenteral , Infecciones del Sistema Respiratorio/inmunología , Virosis/inmunología , Animales , Peso Corporal , Ganglios Linfáticos/citología , Ganglios Linfáticos/inmunología , Masculino , Ratones , Líquido del Lavado Nasal/citología , Líquido del Lavado Nasal/inmunología , Cuello , Factores de TiempoRESUMEN
OBJECTIVE: To determine the influence of route of nutrition on gut mucosal addressin cellular adhesion molecule-1 (MAdCAM-1) expression and the effect of MAdCAM-1 blockade on gut-associated lymphoid tissue (GALT) lymphocyte populations and established respiratory antibacterial immunity. SUMMARY BACKGROUND DATA: Lymphocytes, sensitized to antigens in Peyer's patches, migrate via mesenteric lymph nodes and home to intestinal lamina propria. MAdCAM-1 located on endothelial cells regulates this trafficking. Experimentally, parenteral nutrition (PN) decreases GALT cell mass and mucosal immunity when compared with enteral feeding. METHODS: In experiment 1, MAdCAM-1 expression was quantified in 32 mice after 4 days of feeding chow, a complex diet, intragastric (IG)-PN, or PN. In experiment 2, MAdCAM-1 was measured in 102 mice 0, 4, 8, 12, 24, 48, or 72 hours after starting PN and at 0, 4, 8, 12, 24, or 48 hours after reinstituting chow following 5 days of PN. In experiment 3, 56 mice received chow, PN, chow + MECA-367 (anti-MAdCAM-1 mAb), or chow + Isotype control Ab (IsoAb) for 5 days, followed by Peyer's patches, lamina propria, and intraepithelial lymphocyte yield with respiratory and intestinal IgA levels. In experiment 4, 10 days after Pseudomonas immunization, mice received chow + MECA-367 or chow + IsoAb for 4 days followed by 1.2 x 108 Pseudomonas intratracheally. RESULTS: Diet and route affect MAdCAM-1 expression (chow > complex diet > IG-PN > PN). Decreased MAdCAM-1 expression occurred within hours of starting PN in Peyer's patches, but not mesenteric lymph nodes or the intestine, and recovered quickly with enteral refeeding. MAdCAM-1 blockade reduced all GALT populations. Blockade had little effect on IgA levels and partially impaired the late response of established respiratory immunity. CONCLUSIONS: Enteral feeding affects MAdCAM-1 expression. Complete MAdCAM-1 blockade reduces GALT lymphocytes to PN levels, but the chow feeding stimulus preserves IgA and early antibacterial resistance, implying the existence of non-MAdCAM-1 mechanisms to preserve mucosal immunity.