ABSTRACT
Las endotoxinas bacterianas son lipopolisacáridos (LPS) localizados exclusivamente en la membrana externa de las bacterias gramnegativas, su ingreso al organismo a través de productos parenterales puede causar fiebre, taquicardia, aumento de presión sanguínea y en algunos casos ocasiona la muerte. Existen normativas internacionales acerca del límite de endotoxina para productos farmacéuticos inyectables, tal como las inmunoglobulinas (IgG), que pueden estar expuestas a contaminación durante el proceso de producción y por lo tanto es necesario realizar pruebas para la determinación de endotoxinas bacterianas. El método del lisado de amebocitos de Limulus (LAL) es una de ellas.Este método se fundamenta en la reacción del LAL, el cual es un extracto de células sanguíneas que interaccionan con endotoxinas, activando la cascada del proceso de coagulación y originando la formación de la coagulina. En diversas ocasiones algunas proteínas intervienen en la activación o desactivación de esta cascada, bien sea potenciando o inhibiendo la formación del coágulo. En el caso de la potenciación, el calentamiento es uno de los métodos recomendados por la farmacopea estadounidense (USP) para eliminar interferencias, puesto que desnaturaliza las proteínas que causan la potenciación sin pérdida de endotoxinas. En este trabajo se validó la determinación de endotoxinas bacterianas en IgG mediante el método LAL, el cual es un método rápido y de fácilejecución, por lo que puede implementarse como ensayo de rutina en control de calidad y por ende nos permite agilizar las Liberaciones de Lotes de estos productos.
Bacterial endotoxins are lipopolysaccharides (LPS) located exclusively in the outer membrane of gram-negative bacteria, their entry into the body through parenteral products can cause fever, tachycardia, increased blood pressure and in some cases cause death. There are international standards for endotoxin limit for injectable pharmaceuticals, such as immunoglobulins (IgG), which may be exposed to contamination during the production process and therefore it is necessary to test for determination of bacterial endotoxins. The method of the Limulus amebocyte lysate (LAL) is one of them. This method is based on the LAL reaction, which is an extract of blood cells which interact with endotoxin, triggering the cascade of the coagulation process and causing the formation of coagulin. On several occasions some proteins involved in the activation or deactivation of this waterfall, either by enhancing or inhibiting clot formation. In the case of empowerment, the warming is one of those recommended by the US Pharmacopoeia (USP) to eliminate interference, since denatures proteins that cause endotoxin enhancement lossless methods. In this paper the determination of bacterial endotoxins in IgG was standardized by the LAL method, which is quick and easy to implement method, which can be implemented as a routine test in quality control and thus allows us to streamline releases Lots of these products.
Subject(s)
Humans , Male , Female , Immunoglobulins , Endotoxins , Gram-Negative Bacteria , Lipopolysaccharides , Antibody-Producing CellsABSTRACT
Objective] To observe the clinical efficacy of Agkistrodon in treating Sjogren's syndrome. [Methods] 52 patients were randomly divided into two groups: Treatment of 26 patients taking Agkistrodon and the basic prescription medicine(YIQIYANGYINQUYU drugs); 26 patients in the control received the basic prescription medicine. The clinical symptoms,the disease activity score of SSDAI,the score of TCM symptom and life quality and the index of serology were recorded in this clinical observation. [Results] The treatment group had better results of immune function than the control group after treatment, in particular had a substantial decrease in IgG. In addition, the clinical symptoms, the disease activity score of SSDAI, the score of TCM symptom and life quality were improved. [Conclusion] Agkistrodon has significantly immunosuppressive effects on the patients with Sjogren's syndrome , thus improving the patients' condition,the standard of living and the quality of life.
ABSTRACT
@#ObjectiveTo investigate the disruption of blood-brain barrier (BBB) after brain trauma in rats using IgG immunohistochemical staining.MethodsAn impact-acceleration head injury model was established with rats. Histological changes of rats' brains were observed by HE staining and light and electron microscopes at 1 h, 6 h, 24 h and 7 d after injury, and BBB permeability was analyzed semi-quantitatively by IgG immunohistochemical staining at the same time points.ResultsThe spot bleeding and brain edema was present in the damage region after brain trauma and endothelial cell damage and astrocyte swelling could be found under electron microscope. The extravasations of IgG was detected in the injured hemisphere of rats at 1 h, and achieved the peak at 6 h, remained a high level up to 24 h, and decreased at 7th d.ConclusionThe disruption of BBB function occurs after brain trauma in rats, and detection of IgG extravasations tested by immunohistochemical staining is a simple and sensitive way to investigate BBB permeability.