ABSTRACT
The physiological role of C-reactive protein (CRP), the classical acute-phase protein, is not well documented, despite many reports on biological effects of CRP in vitro and in model systems in vivo. It has been suggested that CRP protects mice against lethal toxicity of bacterial infections by implementing immunological responses. In Achatina fulica CRP is a constitutive multifunctional protein in haemolymph and considered responsible for their survival in the environment for millions of years. The efficacy of Achatina CRP (ACRP) was tested against both Salmonella typhimurium and Bacillus subtilis infections in mice where endogenous CRP level is negligible even after inflammatory stimulus. Further, growth curves of the bacteria revealed that ACRP (50 µg/mL) is bacteriostatic against gram negative salmonellae and bactericidal against gram positive bacilli. ACRP induced energy crises in bacterial cells, inhibited key carbohydrate metabolic enzymes such as phosphofructokinase in glycolysis, isocitrate dehydrogenase in TCA cycle, isocitrate lyase in glyoxylate cycle and fructose-1,6-bisphosphatase in gluconeogenesis. ACRP disturbed the homeostasis of cellular redox potential as well as reduced glutathione status, which is accompanied by an enhanced rate of lipid peroxidation. Annexin V-Cy3/CFDA dual staining clearly showed ACRP induced apoptosis-like death in bacterial cell population. Moreover, immunoblot analyses also indicated apoptosis-like death in ACRP treated bacterial cells, where activation of poly (ADP-ribose) polymerase-1 (PARP) and caspase-3 was noteworthy. It is concluded that metabolic impairment by ACRP in bacterial cells is primarily due to generation of reactive oxygen species and ACRP induced anti-bacterial effect is mediated by metabolic impairment leading to apoptosis-like death in bacterial cells.
Subject(s)
Animals , Anti-Bacterial Agents/pharmacology , Apoptosis/drug effects , Bacillus subtilis/drug effects , Bacillus subtilis/metabolism , C-Reactive Protein/isolation & purification , C-Reactive Protein/pharmacology , Gluconeogenesis/drug effects , Glycolysis/drug effects , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/metabolism , Gram-Negative Bacterial Infections/microbiology , Hemolymph/metabolism , Homeostasis/drug effects , Immunoblotting , Lipid Peroxidation/drug effects , Male , Mice , Oxidation-Reduction , Reactive Oxygen Species/metabolism , Salmonella Infections/drug therapy , Salmonella Infections/metabolism , Salmonella Infections/microbiology , Salmonella typhimurium/drug effects , Salmonella typhimurium/metabolism , SnailsABSTRACT
C-reactive protein (CRP) is found to be a normal component of serum of freshwater air-breathing murrel Channa punctatus. Based on the property of binding with C-polysaccharide (CPS) of pneumococcus bacteria in presence of Ca2+, CRP was purified by phosphorylcholine-Sepharose affinity column chromatography. Molecular weight of the intact protein molecule was estimated to be approximately 141,000 by gel filtration. In non-reduced and reduced conditions the molecule showed molecular weight approximately 28,000 and 14,000 respectively in SDS-PAGE. Monospecific antisera was raised against the affinity purified CRP and used as a tool to detect CRP in the hepatic cytosol and egg extract. The level of CRP in the normal serum was estimated to be 220 micrograms/ml.
Subject(s)
Animals , C-Reactive Protein/isolation & purification , Chromatography, Gel , Electrophoresis, Polyacrylamide Gel , Fishes , Immunodiffusion , Immunoelectrophoresis , RadioimmunoassayABSTRACT
Los avances tecnológicos de las últimas décadas han rescatado pruebas de gran utilidad clínica como la determinación cuantitativa de los reactantes de fase aguda, dentro de los cuales, uno de los más interesantes, desde el punto de vista clínico, es la proteína C reactiva (PCR). La variada tecnología disponible del laboratorio clínico, permite al médico mediante la cuantificación rápida y confiable de los niveles de PCR, se mejore el diagnóstico y manejo de la mayoría de los procesos infecciosos y/o necróticos. En este artículo se analizan las diferentes situacuiones clínicas en las que la determinación seriada de la PCR ha demostrado ser de gran utilidad práctica dentro de las cuales las más importantes son en la cirugía de alto riesgo de infección (cardiovascular, Abdominal, de cadera y neurológica), en el manejo de enfermedades infecciosas de origen bacteriano, particularmente en inmunosuprimidos, en las enfermedades reumatológicas y en el infarto del miocardio. Existen además muchas otras indicaciones clínicas.