Diagnostic values of gene expression of cytokeratin - 19 mrna and mammaglobin mrna in breast cancer patients using real-time polymerase chain reaction

Breast cancer is the most frequent cancer in women. It constitutes almost 20% of all malignancies in women. Currently it affects approximately 6% of the female population. Even before clinical detection of a primary tumour, cancer cells can invade the adjacent structures from where they travel through lymphatic and blood vessels as circulating tumour cells [CTCs]. CTCs colonize distant organ sites as disseminated tumour cells [DTCs] and eventually form microscopic deposits [micrometastasis < 2 mm in diameter], which may remain dormant, but then ultimately lead to an overt metastatic disease. Cytokeratins [CKs] have become the most widely accepted protein markers for the detection of epithelial tumour cells in mesenchymal tissues, BM, blood and lymph nodes. Based on its breast cancer-association and somewhat unique breast-specific pattern of expression, mammaglobin was believed to be an excellent candidate for a novel and clinically useful breast tumor marker, especially in detecting micrometastasis. This study was performed on one hundred female Individuals. They classified into: Group I: 20 apparently healthy females as control group. Group II: 20 females with stage I breast cancer . Group HI: 20 females with stage II breast cancer. Group IV: 20 females with stage III breast cancer. Group V: 20 females with stage IV breast cancer. The following specific investigations were done for all the studied persons:-Cancer Antigen 15-3 [CA15-3] and Carcinoembryonic Antigen [CEA] using chemilmnmescent immunometric assay [IMMULITE 1000 Analyzer]. Cytokeratin-19 mRNA [CK-19] mRNA and mammaglobin mRNA by Real-time polymerase chain reaction [RT-PCR]. In group I [control group]: All the 20 healthy control females had low expression values for CK-19 and Mammaglobin. In group 2 [stage I breast cancer]: 35% of patients had over expression values for CK-19 and 20% had over expression values for Mammaglobin. In group 3[stage II breast cancer]: 47.4% of patients had over expression values for CK-19 and 47.4% had over expression values for Mammaglobin. In group 4 [stage III breast cancer] 68.4% of patients had over expression values for CK-19 and 73.7% had over expression values for Mammaglobin. In group 5 [stage IV breast cancer]: 95% of patients had over expression values for CK-19 and 95% had over expression values for Mammaglobin. Our findings support that all patients with breast cancer should be evaluated by CK-19 and Mammaglobin as a regular laboratory assessment beside the routine tumour markers specially in early stages of breast cancer to detect CTCs at the time of diagnosis

New pH-Metric method for determination of some angiotensin converting enzyme inhibitors through their complex formation with some transition metal ions

Study of mixed ligand complexes of Co[2+] and Zn[2+] with three Angiotensin converting enzyme inhibitors [ACE] was conducted in aqueous solution by pH metric titration. These are Enalapril maleate [I], Perindopril [II] and Moexipril [III] and one of the amino acid family [glycine]. Protonation constants of the three drugs and stability constants of the binary and ternary complexes were determined at [25 +/- 1] [degree sign] C and ionic strength 0.1M NaCl. The stability constants of the binary complexes of I, II and III with Co[2+] and Zn[2+] were performed and the results indicate the presence of one formation constant for the complexes of Co[2+] and Zn[2+] ions with II and III and two formation constants for I. By introducing glycine in the above binary complexes, ternary complexes were formed. The formation constant of the ternary complexes of I, II and III with Co[2+] and Zn[2+] ions with glycine was also calculated at the same temperature and ionic strength. This potentiometric technique was utilized also for the determination of the cited drugs

Spectrophotometric determination of ACE inhibitor drugs through ion-pair formation

Two simple and sensitive spectrophotometric methods were described for the determination of some angiotensin converting enzyme inhibitor drugs [ACE] such as Enalapril maleate [I], Perindopril [II], and Moexipril [Ill]. The two methods involved the formation of ion-pair reactions. The first method was based on the interaction of the cited drugs with bromothymol blue [BTB] at pH 2.5 followed by extraction by methylene chloride which absorbed at 404 nm for I, III and 415 nm for II. The second method was based on the reaction of molybdenum [V]-thiocyanate with the drugs followed by extraction of the complexes formed by methylene chloride which absorbed at 468 nm. The optimum conditions for the ion-pairs formation were established. Beer's law was obeyed for the first method over concentration ranges of 10-48, 4-40 and 5-45 micro g ml[-1] for I, II and III; while for the second method, it was obeyed over concentration ranges of 10-80 micro gml [-1] for l and II and 1-60 micro gm[-1] for III, respectively. The relative standard deviation calculated for Beer's law concentrations, were found to be less than +/- 1% for both methods. The limits of detection [LOD] calculated from the calibration curves in the first method were 0.4, 0.1 and 0.3 micro gm[-1] for I, II and III; respectively; whereas in the second method were 0.18, 0.2 and 0.35 micro gml[-1] for I, II and III, respectively. The two methods could be applied successfully for the determination of the three drugs in tablets, with no evidence of interference from excipients

Spectrophotometric microdetermination of Mo [V] in pure and in steel using its thiocyanate complex interaction with chloroquine and pyrimethamine reagents

A rapid simple, accurate and sensitive spectrophotometric method was developed for the determination of a low concentration of molybdenum in pure and in complicated matrix. It is based on the extraction of the ion-pairs of Mo [V]-thiocyanate-chloroquine [1] and Mo [V]-thiocyanate-pyrimethamine [2], which are orange-red in color into dichloromethane from hydrochloric acid medium. The ion-pairs have an absorption maximum at gamma = 467 and 471 nm with molar absorptivities [epsilon] of 6.16 x 103 and 5.80 x 103 1/mol/cm for the Mo [V] ion- pairs [1] and [2]. The Sandell sensitivity was found to be 0.027 and 0.042 mug/cm2 for [1] and [2]. The effect of temperature, reagent concentration, time and diverse ions upon the absorbance of the ion- pairs was critically assessed. Beer's law was valid over a concentration range of 2.0-22.0 and 2.0-25.0 mug Mo/ml in case of [1] and [2]. Both reagents [chloroquine and pyrimethamine] were successfully applied for the determination of molybdenum in synthetic mixtures and molybdenum steels

Elisa in detection of entamoeba histolytica antigens in stools

The modified double antibody sandwish ELISA was applied to detect E. histolytica antigens in the stool samples of 20 patients have amoebiasis [symptomatic or asymptomatic], 10 with intestinal parasites other than E. histolytica and 10 healthy individuals. The method was found to be sensitive [90%] and specific [85%]. It is concluded that ELISA could be applied for a large scale epidemiological study and immunodiagnosis of amoebiasis