Proteomic analysis of plasma proteins after low-level laser therapy in rats.

The laser radiation absorbed by cells induces production of reactive oxygen species (ROS), followed by the development of oxidative stress. Proteins are major targets for ROS due to their abundance in biological systems. The aim of the present pilot study was to examine the effects of transcutaneous laser blood irradiation (TLBI), i.e., low-level laser therapy (LLLT) at 830 nm on plasma proteome in Wistar rats. Rats were irradiated in the heart area (i.e. coronary arteries) daily (i.e., for 9-day period), by commercially available GaAsAl diode laser (Maestro/CCM, Medicom Prague, Czech Republic, lambda=830 nm, power density 450mW/cm(2), daily dose 60,3 J/ cm(2), irradiation time 134 sec). The comparison of blood plasma proteome from irradiated and non-irradiated rats was performed utilizing 2D electrophoresis followed by MALDI TOF/TOF mass spectrometry. LLLT led to a quantitative change in the acute phase proteins with antioxidant protection i.e., haptoglobin (log(2) fold change (FC)=3.5), hemopexin (log(2) FC=0.5), fibrinogen gamma (log2 FC=1.4), alpha-1-antitrypsin (log(2) FC=-2.2), fetuin A (log2 FC=-0.6) and fetuin B (log2 FC=-2.3). In comparison to conventional biochemical methods, the changes in protein levels in blood plasma induced by LLLT offer a deeper insight into the oxidative stress response.

Differential Urinary Proteomic Analysis of Endometrial Cancer.

Endometrial cancer is one of the most frequent gynecological malignancies present in more than 95 % of all uterine cancers. In spite of that, screening of such disease is not commonly performed in clinical practice due to enormous costs and relatively low sensitivity. Therefore, developing an effective screening test to diagnose endometrial cancer at early stages is of great importance for the clinical area of investigation. In this work, we applied urinary proteomics (i.e., bottom-up proteomic approach followed by nano HPLC-ESI-MS/MS) in patients with endometrial cancer, with respect to find proteins aimed for the early diagnostics and screening. According to the results, the significant semi-quantitative changes were observed in urinary proteome of treated patients. The proteins that may be pivotal in pathogenesis of endometrial cancer, like cadherin-1 (CDH1), vitronectin (VTN) and basement membrane specific-heparan sulphate proteoglycan core protein (HSPG2) were down-regulated, when compared to the control group. Ultimately, it can be stated that urinary proteomics has a potential for the searching of cancer protein biomarkers based on their altered concentration.

Value of Ki67 immunostain in identification of malignancy in serous effusions.

The distinction between malignant and benign serous effusions continues to be a challenging and a frequent problem to cytopathologists. Recently, immunostains employing various antibodies have improved the diagnostic accuracy of malignant effusions. We investigated the usefulness of Ki67 (MIB1) antigen immunostaining in the evaluation and diagnosis of malignant serous effusions. Cell block sections from a total of 54 cases of serous effusions cytologically diagnosed as malignant (28), suspicious (6), and benign (20) were immunostained with MIB1 monoclonal antibody to the Ki67 nuclear proliferation antigen according to the avidin-biotin immunoperoxidase method. The patients were 30 women and 24 men with an average age of 58 yr. Ki67 (MIB1) immunostain labeling index (LI) values were higher than 20% in 23 of 28 (82%) cytologically malignant, in 3 of 6 (50%) suspicious, and in 1 of 20 (5%) benign/reactive. Further investigation revealed histologic, radiologic, and/or clinical evidence of malignancy in the 3 suspicious (but not in the benign/reactive) cases with Ki67 LI values higher than 20%. Correlation between Ki67 LI (> 20%) and cytologic effusion type (benign, suspicious, or malignant) was statistically significant (P < 0.0001). Ki67 immunostaining has value as an adjunct testing to cytomorphology and other immunostains in distinguishing benign from malignant effusions. The addition of Ki67 immunostaining to conventional cytology appears more sensitive than cytomorphology alone and may assist in arriving at accurate diagnoses in suspicious cases with inconclusive cytomorphologic features.

Improved detection of adenocarcinoma of serous fluids with p53 immunocytochemistry.

OBJECTIVE: To investigate the potential value of p53 protein immunostaining in identifying malignant cells in serous fluids. STUDY DESIGN: We applied p53 immunostaining to 26 cytologically malignant, 8 suspicious and 34 benign specimens of serous fluids from 68 patients. For comparison, staining for carcinoembryonic antigen (CEA) was also done on all the specimens. RESULTS: CEA was positive in 23 of 26 (88%) cytomorphologically malignant, 3 of 8 (38%) suspicious and 1 of 34 benign cases. p53 Nuclear immunostaining was positive in 12/26 (46%) malignant, 2/8 (26%) suspicious and no benign cases. Correlation between p53 staining and serous fluid type (benign, suspicious or malignant) was significant. The P based on Fisher's exact test was < .0001. Two cases that were reported cytomorphologically as suspicious stained positively with p53; further investigation in those cases confirmed the diagnosis of metastatic adenocarcinoma. CONCLUSION: p53 Immunostaining of serous fluids seems to be of value in identifying carcinoma cells, especially in those cases that show inconclusive or bland cytologic features. Combining p53 with CEA immunostains in clinically or cytologically suspicious cases may assist in recognition of carcinoma cells and in pursuing an appropriate therapeutic approach.

P53 protein immunohistochemical expression in colonic adenomas with and without associated carcinoma.

OBJECTIVES: P53 protein immunohistochemical (IHC) expression was investigated in a series of colonic adenomas and carcinomas to determine the p53 immunohistochemical expression of adenomas in general compared with carcinomas, the difference in staining pattern between adenomas with associated carcinoma and those without associated carcinoma, and the difference in p53 staining in the usual adenomas (low-grade dysplasia) compared with those harboring high-grade dysplasia. METHODS: The study involved a series of 20 adenomas without concurrent carcinoma (group 1 adenoma), 29 adenomas with concurrent carcinoma (group 2 adenoma), and 20 carcinomas. Sections of the paraffin-embedded tissues were stained with DO-7 p53 monoclonal antibody after microwave antigen-retrieval method. Cases with nuclear staining in > or = 20% of the tumor cells were considered positive. RESULTS: Analysis of results showed that 65% of carcinomas and 37% of all adenomas were reactive with p53 IHC staining (p = 0.03). With respect to the adenomas, 30% of group 1 and 41% of group 2 adenomas were reactive for p53 protein (p = 0.42). CONCLUSIONS: Our data demonstrate a statistically significant higher p53 expression rate in colonic carcinomas than in adenomas, and that adenomas with concurrent carcinomas are more frequently p53 positive than those without concurrent carcinoma, but this was not statistically significant. Also, p53 expression is more frequent and intense in adenomas with high-grade dysplasia (10/20, 50%) than in ordinary adenomas with low-grade dysplasia (8/29, 28%), which suggests a strong correlation between the degree of dysplasia in colonic neoplasia and p53 expression pattern.