Serum trace elements in diagnosis of breast malignancy

Breast cancer is a highly heterogeneous disease globally. The absolute risk of breast cancer increases with age and is higher in postmenopausal women. This study tried to find out role of serum trace elements [cadmium, cobalt, copper and zinc] and biomarkers in diagnosis of breast malignancy. Levels of serum trace elements were estimated by atomic absorption spectrophotometry in serum samples of 23 breast cancer patients of stage III. Analysis of protein pattern was observed by sodium dodecyl polyacrylamide gel electrophoresis [SDS-PAGE]. In menstruating women, the mean serum level of cadmium, copper and cobalt were significantly lowered as compared to control, however, zinc showed no significant difference. In premenopausal and postmenopausal women, the mean serum level of cadmium, copper, cobalt and zinc was decreased significantly in patients as compared to controls [p<0.001]. The ratio of Cu/Zn was also decreased in patients when compared to controls. Three major peptide bands with molecular weights of 33 kDa, 52 kDa and 185 kDa appeared in these samples when subjected to SDS PAGE. Serum trace elements are significantly lower in breast cancer patients compared to controls. The exact mechanism responsible for the alterations in trace element levels in patients with breast cancer is unclear and requires further evaluation. Gel electrophoresis may be important to find out breast cancer markers that may help in diagnosing the disease and response to treatment.

Screening of pneumococcal pneumonia by amplification of pneumolysin gene in children visiting hospitals in Lahore, Pakistan

Streptococcus pneumoniae is a common worldwide potential pathogen causing pneumonia among children and the detection of pneumococcal infections by conventional culturing techniques is cumbersome. The present study describes a comparative analysis of sensitive nested-PCR and bacterial culture in pediatric patients with clinical and radiological indication of S. pneumoniae infection. PCR was performed using outer primers to amplify a 348-bp region and inner primers a 208-bp region of the pneumolysin gene. For pneumolysin PCR assay, DNA from peripheral blood and middle ear fluid [MEF] samples was extracted by salting out method. The sensitivity of the assay was evaluated with about 0.06 pg of purified S. pneumoniae genomic DNA. Among 90 MEF culture negative samples from acute otitis media pediatric patients, 8.8% pneumolysin-PCR positivity was detected, demonstrating the sensitivity and reliability of PCR for rapid pneumonia evaluation. Binomial test of proportionality performed on [SPSS 17] gives P<0.05 indicating that PCR technique is statistically significant and sensitive in the diagnosis of S. pneumoniae infection. The research work evaluated the effectiveness and efficacy of nested-PCR for detecting S. pneumoniae in pediatric patients with clinical and radiological confirmation of bacterial infection. This simplified method permitted quick selection of the patients and played a significant role in preliminary management of pneumococcal infections

Alpha1-microglobulin: a marker for early detection of tubular disorders in diabetic nephropathy

Tubular damage as suggested by tubular proteinuria is a recognised feature of glomerulonephritis. The objectives of the study were to compare the level of alpha-microglobulin in normal and diabetic patients, and also to find out whether the level of alpha-1 microglobulin could become a laboratory marker for tubulo-interstitial damage in diabetic nephropathy. Twenty-nine registered Type II diabetic patients of either sex were studied. The patients' age ranged from 41-50 years who were admitted in the medical ward, and those who visited the outdoor department of Sir Ganga Ram Hospital, Lahore were included in the study. The duration of study was one year from June 2006 to June 2007. Ten normal subjects with no history of diabetes were taken as controls. Blood samples and 24 hour urine samples of patients of all groups were collected. The levels of urinary protein and blood sugar were estimated by auto analyser. Proteinuria positive urinary samples were analysed by SDS-PAGE electrophoresis. The level of alpha-1 urinary protein was significantly increased in the group of diabetic patients as compared to the urinary protein level of normal subjects. Blood sugar level was also significantly increased in patients as compared to controls. Level of low molecular weight protein alpha-1 microglobulin showed an electrophoresis band of 28 Kda with an average volume of 6741.88 in the urine sample of patients. On the other hand, a very light, hardly recognizable band was observed in normal subjects. Urinary alpha-1 microglobulin provides a noninvasive and inexpensive diagnostic alternative for the diagnosis and monitoring of urinary tract disorders, i.e., early detection of tubular disorders of diabetic nephropathy. We propose that SDSPAGE electrophoresis is a comparatively inexpensive diagnostic approach to detect this marker in the urine sample

Diagnostic significance of adenosine deaminase in pleural tuberculosis

Tuberculosis [TB] is a major cause of pleural effusion, which in TB usually has lymphocytic and exudative characteristics. The analysis adenosine deaminase [ADA] activity is a very useful diagnostic approach to achieve a more rapid and precise diagnosis in the cases of pulmonary TB. Fifty male and fifty female patients presenting with tuberculous pleural effusion were included in the study. The patients were taken from the medical ward of Sir Ganga Ram Hospital. Activity of adenosine deaminase [ADA] was estimated by the technique of Sodium dodecyl sulphate electrophoresis [SDS-EF] using 10% polyacrylamide gel. There is an increased level of enzyme ADA in pleural fluid of tuberculous patients as compared to normal subjects. Estimation of ADA activity may provide the basis for the rapid and efficient diagnosis of pleural TB in different clinical settings. However study should be carried out on large number of patients to reach a better conclusion

Electrophoretic analysis of serum proteins in prostate cancer

Objective: This work was undertaken to analyse qualitatively serum proteins separated by SDS-PAGE and stained with Coomassie Brilliant Blue R-250, to detect novel proteins, which could possibly serve as markers for the early detection of prostate cancer

Design: This study [carried out on subjects who were confirmed by biopsy as cancer prostate [CaP] patients] was a preliminary step towards the detection of some new protein markers for CaP

Place and Duration of Study: This study was conducted at the Institute of Biochemistry and Biotechnology University of the Punjab Lahore, from January to March 2003

Subjects and Methods: Serum samples, 36 cases of cap and 36 controls of similar age group were separated by SDS-PAGE on a 12% gel, and then stained using Coomassie Brilliant Blue R-250. Protein fractions were analyzed using the computer software program "GeneGenius Gel Documentation and Analysis System"

Results: The appearance and relative raw volumes of 14 major protein fractions ranging in molecular weight from 0.23-157 kD in each group were studied. It was seen, on the whole, that the raw volume of most of the protein fractions decreased in the CaP cases as compared to the controls. Furthermore, the protein fractions of 1.27, 100, 114, 122 and 140 kD were absent in a significant number of the cases. However, it was seen that the protein fraction of 0.23 kD was absent in all the control samples while it appeared in most of the cases

Conclusion: A number of proteins were found to have been lost during malignant transformation. Further investigations are warranted to identify these novel proteins using 2D-PAGE followed by immunoblotting