Evaluation of migraine in patients with ischaemic stroke.

An association between migraine and ischaemic stroke has been observed for many years, but the exact mechanisms by which migraine can lead to stroke are still unknown. The purpose of this study was to determine the prevalence of migraine headaches in patients with ischaemic stroke. In this prospective cohort study, we assessed 323 patients with ischaemic stroke; these diagnoses were assigned based on the International Headache Society criteria for migraine with or without aura. Patients were recruited without major risk factors such as stroke, hypertension, hyperlipidaemia, diabetes, taking oral contraceptive pills, history of drug abuse and trauma in issue of their Stroke. Data were collected via a written questionnaire upon admission and were analysed with SPSS version 16 software. Comparisons were performed using Mann-Whitney's U test and chi-square and t-test. Migraine headache was present in 11.2% (36 of 323) of patients, 8.1% of women and 3.1% of men. Migraine prevalence was highest in the age over 60  years. There was a history of migraine without aura for over 2  years in 6.2% of patients with ischaemic stroke. Also, we found no significant correlation between migraine headache and location of the lesion in patients with ischaemic stroke.

Preparation of antibody against horseradish peroxidase using hybridoma technology.

BACKGROUND: Monoclonal antibody against horseradish peroxidase (HRP) has many applications which peroxidase anti-peroxidase. Peroxidase-antiperoxidase (PAP) complex formation is its most known and important usage. This complex is used in many immunohistochemical and immunocytochemical staining techniques. OBJECTIVE: The aim of this study was the preparation of anti-HRP monoclonal antibody through hybridoma technology. METHODS: The BALB/c mice were immunized by repeated injections of HRP. After the confirmation of their immunization by ELISA test, the spleen lymphocytes and SP2/0 myeloma cells were hybridized using PEG as fusing agent. The hybridoma cells were then selected by culturing in HAT medium. Identification and selection of anti-HRP producing clones were done by ELISA test on culture supernatants of the obtained clones. To acquire the monoclones, limiting dilution was performed twice and the effect of finally obtained antibodies on enzyme activity was investigated by a specific ELISA test. In vivo tumor induction method was used for production of concentrated antibody. At last class and subclass of the obtained antibodies were determined by Isostrip Kit. RESULTS: After seven rounds of cell fusions, 224 clones were obtained, from which, six ones were anti-HRP producers. Two clones (P1F11 and P2F6) with higher antibody secretion were selected and subcloned. Both derived hybridoma monoclones (P1F11D2 and P2F6F3) were producing antibodies from IgG1 subclass with kappa (Kappa) light chains which didn't affect the enzyme activity. The electrophoresis of ascetic fluid of tumor induced mice showed an obvious band in gamma (gamma) position. CONCLUSION: The obtained monoclonal antibodies are from IgG class and don't affect the enzyme activity, therefore it seems that they are suitable for PAP complex production.

Production of monoclonal antibody against alkaline phosphatase.

BACKGROUND: Monoclonal antibody against alkaline phosphatase (Alp) has many applications which Alp-anti Alp complex (APAAP) formation is one of the most important ones. This complex is applicable in many immunohistochemical and immunocytochemical techniques such as diagnosis of various kinds of leukemias, lymphomas, skin diseases, kidney dysfunctions, etc. OBJECTIVE: Production of anti-Alp monoclonal antibody for utilization in APAAP complex. METHODS: After several arranged injections of Alp to Balb/c mice and determining the specific antibody titer by ELISA test, the spleen lymphocytes of immunized mice and Sp2/0 myeloma cells were fused using polyethylene glycol as fusing agent and hybridoma cells were cultured in HAT medium. Identification and selection of anti-Alp producing clones were done by performing ELISA test on supernatants of all resulting clones. Limiting dilution method was used to attain monoclones and the effect of obtained antibodies on enzyme activity was investigated by a specific ELISA test. For production of concentrated Ab, the hybridoma cells were injected to peritoneal cavity of mice and the produced ascetic fluids were collected. Finally class and subclass of the obtained antibodies were determined by Isostrip kit. RESULTS: After six rounds of fusion, 104 Hybridoma clones were obtained and two Anti-Alp producing clones (A_1G_8 and A_1G_9) were selected and subcloned. Both antibodies were IgG_1 with kappa (kappa) light chains. These antibodies did not affect the enzyme activity and the electrophoresis of ascetic fluids showed an obvious band in gamma (gamma) position. CONCLUSION: Because these antibodies are from IgG class and don't affect the enzyme activity, it seems that they are suitable for APAAP complex formation.