RESUMEN
OBJECTIVE: The host response to pulmonary Mycobacterium tuberculosis (Mtb) infection results in granuloma formation in an effort to limit infection, but the host immune cells also provide an environment in which Mtb persists. Granuloma formation requires immune cell infiltration and concurrent extensive remodeling of pulmonary tissue which we hypothesize to be the result of increased matrix metalloproteinases (MMP) activity. DESIGN: C57BL/6 mice infected with virulent Mtb (H37Rv) via intratracheal inoculation were treated with a synthetic inhibitor of MMP activity (BB-94). Mice were assessed for colony forming units, granuloma morphology, leukocyte recruitment and cytokine levels over 90 days of infection. RESULTS: BB-94 treated mice had significantly decreased numbers of pulmonary and blood-borne Mtb early during disease, increased collagen deposition within early granulomas and significantly decreased pulmonary leukocyte recruitment when compared to vehicle-treated, Mtb-infected mice. Cytokine expression did not differ significantly between groups. CONCLUSION: Events of early granuloma formation can be modified by inhibiting MMP activity, by decreasing leukocyte recruitment, a major source of MMPs during infection, enhancing the establishment of granulomas and decreasing blood-borne dissemination of Mtb.
Asunto(s)
Granuloma/patología , Inhibidores de la Metaloproteinasa de la Matriz , Fenilalanina/análogos & derivados , Fenilalanina/administración & dosificación , Tiofenos/administración & dosificación , Tuberculosis Pulmonar/patología , Animales , Recuento de Colonia Microbiana , Citocinas/análisis , Esquema de Medicación , Femenino , Recuento de Leucocitos , Pulmón/enzimología , Pulmón/patología , Ratones , Ratones Endogámicos C57BL , Tuberculosis Pulmonar/enzimologíaRESUMEN
The aim of this study was to investigate matrix metalloproteinase (MMP) expression during the immune response to pulmonary Cryptococcus neoformans (Cne) infection. The immune response generated in C.B-17 and C57BL/6 mice to pulmonary Cne infection has previously been characterized as type 1 and type 2, respectively, differing in the cytokines produced and leukocytes recruited during infection, influencing the extent of Cne clearance from the lung. The focus of this study was to examine changes in expression of MMP-2 and tissue inhibitor of metalloproteinase (TIMP)-2 in the lungs of Cne-infected mice during the two types (type 1 vs. type 2) of responses. C.B-17 mice that formed well-defined granulomas had elevated levels of pulmonary MMP-2 mRNA early during infection. C57BL/6 mice that had poorly defined cellular aggregates did not express detectable levels of pulmonary MMP-2 mRNA until later in the infection. Specific expression of MMP/TIMP was correlated with the type of immune response present, resolution of Cne infection and the resulting lung pathology.
Asunto(s)
Criptococosis/microbiología , Cryptococcus neoformans , Granuloma del Sistema Respiratorio/microbiología , Metaloproteinasa 2 de la Matriz/biosíntesis , Animales , Criptococosis/inmunología , Criptococosis/metabolismo , Modelos Animales de Enfermedad , Granuloma del Sistema Respiratorio/inmunología , Granuloma del Sistema Respiratorio/metabolismo , Recuento de Leucocitos , Pulmón/inmunología , Pulmón/metabolismo , Metaloproteinasa 2 de la Matriz/genética , Ratones , Ratones Endogámicos C57BL , ARN Mensajero/análisis , Factores de Tiempo , Inhibidor Tisular de Metaloproteinasa-2/biosíntesis , Inhibidor Tisular de Metaloproteinasa-2/genéticaRESUMEN
There is a causal relationship between obesity-associated diabetes and an increased risk of infection. The ability of obese (fa/fa) Zucker rats, a model for non-insulin-dependent diabetes mellitus (NIDDM), to clear Candida albicans from the circulation and tissues was compared to that of lean (Fa/fa, Fa/Fa) Zucker rat controls as a measure of immune function. The ID50 necessary to establish tissue colonisation in lean Zucker rats was 1.18 log10 times greater than that determined for the obese Zucker rats. Nine days after intravenous (i.v.) injection of a yeast suspension, the organs of obese rats had a 10-fold greater yeast/g organ burden than did lean rats. The kidney was determined to be the primary target organ for colonisation. Germ-tube formation by C. albicans occurred at a rate 1.5 times faster in serum from obese rats than in serum from lean rats. Peritoneal polymorphonuclear leucocytes, resident macrophages and thioglycollate-elicited macrophages from lean Zucker rats displayed a significantly higher ability to kill ingested yeast cells than analogous cell populations from obese Zucker rats.
Asunto(s)
Candidiasis/inmunología , Diabetes Mellitus Tipo 2/inmunología , Modelos Animales de Enfermedad , Neutrófilos/inmunología , Ratas Zucker , Animales , Glucemia/análisis , Candida albicans/crecimiento & desarrollo , Candida albicans/inmunología , Candida albicans/fisiología , Candidiasis/complicaciones , Diabetes Mellitus Tipo 2/complicaciones , Susceptibilidad a Enfermedades , Inmunocompetencia , Riñón/microbiología , Riñón/patología , Macrófagos Peritoneales/inmunología , Masculino , Obesidad/complicaciones , Fagocitosis , Ratas , Triglicéridos/sangreRESUMEN
BACKGROUND: To further characterize selected pathologic features on a biochemical level, the authors analyzed the nuclear magnetic resonance metabolite and phospholipid spectra of 30 malignant colon tumors using 31P magnetic resonance spectroscopy. METHODS: Eleven individual generic phospholipids were identified in the spectra of 17 phospholipid extracts, and 31 individual phosphatic metabolites were identified in the spectra of 13 perchloric acid extracts. The metabolites and lipids were quantified for statistical intergroup comparisons based on tumor stage, lymph node status, differentiation, mucin production, blood vessel invasion (BVI), and lymphatic vessel invasion (LVI). RESULTS: Significant elevations in the relative concentration of alpha-glycerol phosphate were noted when comparing AJCC tumor classification (T3 vs. T2, 0.92 +/- 0.14 vs. 0.46 +/- 0.11, P < 0.009), tumor differentiation (moderately vs. well differentiated, 0.92 +/- 0.14 vs. 0.46 +/- 0.11, P < 0.009), and BVI (presence vs. absence, 1.03 +/- 0.04 vs. 0.68 +/- 0.10, P < 0.028) by elastic tissue stain. Among the tissue phospholipids analyzed, the relative concentration of a choline phospholipid was significantly different when comparing moderately and poorly differentiated tumors (6.26 +/- 0.56 vs. 3.29 +/- 0.30, P < 0.001), T2 and T3 tumors (3.90 +/- 0.45 vs. 6.31 +/- 0.56, P < 0.009), and mucin-positive vs. mucin-negative tumors (4.46 +/- 0.56 vs. 6.83 +/- 0.76, P < 0.028). Differences in lymph node status of the cases analyzed in this study (lymph node positive vs. lymph node negative) were noted for tumors with elevated levels of sphingomyelin (8.13 +/- 0.40 vs. 6.88 +/- 0.16, P < 0.02), diminished levels of phosphatidylinositol (5.25 +/- 0.27 vs. 6.38 +/- 0.34, P < 0.02), elevated levels of beta-glycerol phosphate (5.30 +/- 0.70 vs. 1.20 +/- 0.06, P < 0.05), and elevated levels of glycerol 3-phosphoserine (0.48 +/- 0.01 vs. 0.23 +/- 0.02, P < 0.002). CONCLUSIONS: The characteristic differences in the phospholipid and intermediate phosphate metabolite profiles identified through magnetic resonance spectroscopic and histopathologic analysis may provide important information regarding the nature of tumor and cell membrane metabolism. Differences in these profiles may identify markers useful for biologic behavior, provide prognostic information, and characterize the impact of the pathologic features of colon cancer.
Asunto(s)
Neoplasias del Colon/metabolismo , Fosfatos/metabolismo , Fosfolípidos/metabolismo , Diferenciación Celular , Neoplasias del Colon/patología , Humanos , Metástasis Linfática , Espectroscopía de Resonancia Magnética , Invasividad Neoplásica , PronósticoRESUMEN
Saponified phospholipid extracts of malignant and normal human breast and colon surgical tissue specimens (n = 45) generate characteristic phosphodiester profiles using 31P magnetic resonance (MR) spectroscopy. The resultant 31P MR spectroscopic profiles of the analyzed tissues are used to differentiate malignant from normal. The appearance of an uncharacterized resonance at 0.29 delta in the malignant tissue spectra (50% of breast and 75% of colon specimens) is the most notable qualitative finding. Quantitatively, malignant colon tissues differ from normal colon tissues with depressed levels of phosphatidylserine and elevated levels of glycerol 3-phosphorylglycerol and an index measuring the summation of phospholipid polar head group residues with free hydroxyl groups. Malignant breast tissues have significantly elevated levels of glycerol 3-phosphorylethanolamine and significantly depressed levels of glycerol 3-phosphorylcholine compared to normal breast tissues, reflecting a perturbation in the balance of lipid residues that are the respective breakdown products of phosphatidylethanolamine and phosphatidylcholine. The concentration of the polar head group residues is compared to 31P MR spectroscopic profiles of colon and breast tissue phospholipids, in order to demonstrate the quantitative nature of the technique employed.
Asunto(s)
Adenocarcinoma/química , Neoplasias de la Mama/química , Carcinoma Intraductal no Infiltrante/química , Neoplasias del Colon/química , Espectroscopía de Resonancia Magnética , Fosfolípidos/metabolismo , Femenino , HumanosRESUMEN
Phospholipids of 16 malignant and 11 non-malignant human colon specimens were analyzed using a chloroform-methanol analytical reagent in conjunction with 31P magnetic resonance spectroscopy (MRS) at 202.4 MHz. Sixteen individual generic phospholipids were identified and quantified for statistical intergroup comparisons. Statistically significant elevations in the relative concentrations of lysophosphatidylcholine and phosphatidylcholine plasmalogen were seen in malignant tissues along with significantly depressed levels of sphingomyelin and phosphatidylethanolamine plasmalogen. The malignant and non-malignant tissue groups were further differentiated by the detection of the minor phospholipids, lysophosphatidylcholine plasmalogen, lysophosphatidylethanolamine plasmalogen, lysophosphatidic acid and phosphatidylglycerol exclusively present in the malignant tissues and by significant changes in computed phospholipid metabolic indices that were dominated by choline containing lipids. The 31P MRS methods used represent an advancement over previous protocols for identifying and quantifying major and minor tissue phospholipids making this the first direct study of membrane phospholipids in human colon tissues using 31P MRS. The phospholipid profiles obtained may provide important information regarding the nature of the malignant cell's membrane system and identify markers which may be used to estimate malignant propensity, aggressiveness of disease and provide prognostic information.
Asunto(s)
Adenocarcinoma/química , Colon/química , Neoplasias del Colon/química , Fosfolípidos/análisis , Adenocarcinoma/diagnóstico , Colon/patología , Neoplasias del Colon/diagnóstico , Humanos , Espectroscopía de Resonancia MagnéticaRESUMEN
Phosphatic metabolite profiles of 19 malignant and normal human colon specimens were analyzed by techniques of perchloric acid extraction and 31P magnetic resonance spectroscopy at 202.4 MHz. Thirty-one individual phosphorus-containing intermediates of metabolism were identified and quantified for statistical intergroup comparisons. Elevations in relative concentrations of phosphorylethanolamine, IMP, NADP 2'-P, an uncharacterized resonance at 3.72 delta, glycerol 3-phosphorylcholine, phosphorylated glycans and the nucleoside diphosphosugars were seen in malignant tissues concurrently with reductions in relative concentrations of phosphorylcholine, phosphocreatine (PCr), and ATP. The malignant and normal tissue groups were further characterized and contrasted by computing metabolic indices from spectral data. Significant elevations in phosphomonoesters, glycerolphosphodiesters, the ratio of phosphorylethanolamine/phosphorylcholine, and phosphomonoesters/inorganic orthophosphate were detected in malignant tissues along with significant reductions in the ratios of PCr/inorganic orthophosphate, PCr/ATP, the energy charge of the adenylate system and the tissue energy modulus. These results revealed significant alterations in high energy metabolism, low energy metabolism, and membrane metabolism characteristic of malignant tissues. The reduction in high energy phosphates ATP and PCr was balanced by the net increase in nucleoside diphosphosugar and a shift in equilibrium to metabolism involving low energy phosphomonoesters. The spectral data of the tumors, which were of epithelial origin, demonstrated minor metabolites not previously detected in tissue extract analysis of malignant tissues. Detection of these minor metabolites represents an indirect measurement of phospholipid metabolism in malignant tissues.
Asunto(s)
Neoplasias del Colon/metabolismo , Fosfatos/metabolismo , Membrana Celular/metabolismo , Metabolismo Energético , Epitelio/metabolismo , Humanos , Técnicas In Vitro , Espectroscopía de Resonancia Magnética , Lípidos de la Membrana/metabolismo , Músculos/metabolismo , Fosfolípidos/metabolismoRESUMEN
The kinetic patterns of DNA synthesis in wild-type (RAD+) and rad 52 mutants of yeast, which exhibit high levels of synchrony during meiosis, are comparable. However, RAD 52 mutants accumulate single-strand breaks in parental DNA during the DNA synthesis period. Thus, the product of the RAD 52 gene has a role in meiotic DNA metabolism, as well as in the repair of DNA damage during mitotic growth. The observed breaks may be unresolved recombination intermediates.