Immunoexpression of TTF1 and p63 Differentiates Lung Adenocarcinomas in Sputum Samples.

CONTEXT: Differentiating NSCLC as either adeno or squamous type and identification of Epidermal Growth Factor Receptor (EGFR) mutations is clinically relevant for lung cancer patients for selecting treatment. Thyroid transcription factor-1 (TTF-1) and p63 were demonstrated as useful markers for histologic typing of lung cancer. Mutation and overexpression of EGFR has been reported in a subset of non-small cell lung cancers. If these markers can be validated for the differential diagnosis of adenocarcinoma in a sputum sample itself, it will be highly beneficial for lung cancer patients. AIMS: To evaluate whether immunocytochemical expression of TTF-1, p63, and EGFR proteins in sputum samples can be used for differential diagnosis of lung adenocarcinoma by comparing with that of the corresponding tissue samples. SETTINGS AND DESIGN: Ninety sputum samples and matched tissue samples were used for the study. SUBJECTS AND METHODS: Monolayered smears and cell blocks of sputum and the corresponding tissue samples were immunostained with the standard ABC method. The expression patterns of these markers were analyzed statistically and compared with clinic-pathological parameters. STATISTICAL ANALYSIS USED: Chi-square test and paired t-test. RESULTS: The p63 protein had a positive expression in 73.9% of SCC whereas TTF1 had positive expression in 75.8% of ADC. The EGFR expression was positive in 27 cases of adenocarcinoma, 21 cases of SCC and 19 cases of NSCLC. CONCLUSIONS: Immunocytochemistry of the aforementioned antibodies in sputum samples can be used as supplementary evidence for the subtyping of NSCLC.

Immunocytochemistry on Sputum Samples Predicts Prognosis of Lung Cancer.

CONTEXT: Despite sputum cytology being accepted as a simple and noninvasive diagnostic method for lung cancer, the clinical usefulness of sputum for evaluation of prognosis is yet to be explored. Validation of some of the markers in sputum for prognosis prediction will be highly useful for selective therapy. AIMS: This study was aimed to evaluate a reliable panel of immunocytochemical markers for their significance to predict survival. MATERIALS AND METHODS: We have analyzed the expression of p53, p16, galectin-3, and epidermal growth factor receptor (EGFR) proteins in sputum samples processed in a mucolytic agent/cellblock and compared the same with that of the corresponding tissue samples. RESULTS: Overexpression of p16 and EGFR was found to have a better survival benefit, whereas positive p53 and galectin-3 expressions had shorter period of survival. Expression patterns of all these four proteins were more or less similar in smears, cellblocks of sputum, and tissue samples except for slight changes in staining intensity which was not found to be statistically significant. No significant difference was found in the association of these proteins with survival pattern between sputum and tissue samples. CONCLUSION: This is the first report of immunocytochemistry of a panel of markers on cells exfoliated in sputum samples which suggests that analysis of immunocytochemical markers in sputum samples can be attempted as a cost-effective and reliable predictor of prognosis and survival. Accumulation of mutated p53, overexpression of galectin-3, and lower expression of p16 and EGFR proteins were found to predict poor prognosis for lung cancer.

Distribution of water in the pores of periodic mesoporous organosilicates - a proton solid state MAS NMR study.

Solid state proton (1H) magic angle spinning (MAS) NMR has been employed to study the distribution of confined water in ethane substituted periodic mesoporous organosilicate (PMOE) materials. Proton spectra acquired at different hydration levels are analysed and interpreted in terms of water clusters of various sizes and distributions of water layers on the pore surface. For comparison, we also performed similar experiments on SBA-15. The formation of larger clusters at lower hydration suggests that the pores of PMOE are getting filled with water at lower hydration levels than those in SBA-15. For PMOE, the simultaneous presence of two major resonances in the ranges 3.6-4.1 ppm and 4.4-5.2 ppm and their behaviour upon hydration imply a water layer distribution that is the sum of two contributions, corresponding to fully filled and partially filled pores or pore segments. Furthermore, the behaviour mentioned above suggests that both radial and axial filling mechanisms play a significant role in the hydration process. For SBA-15, as a function of hydration, we observed a smooth variation in the proton chemical shift of the main dynamic resonance. In accordance with previous studies, this is attributed to the gradual increase in the average thickness of water layers with an increase in hydration, and to a pore filling mechanism that is predominantly radial.

Significance of DNA Replication Licensing Proteins (MCM2, MCM5 and CDC6), p16 and p63 as Markers of Premalignant Lesions of the Uterine Cervix: Its Usefulness to Predict Malignant Potential

Cervical cancer continues to be a leading cancer among women in many parts of the world. Nation-wide screening with the Pap smear has not been implemented in India due to the lack of adequately trained cytologists. Identification of biomarkers to predict malignant potential of the identified low risk lesions is essential to avoid excessive retesting and follow up. The current study analyzed the expression patterns of DNA replication licensing proteins, proliferation inhibitor protein p16INK4A and tumor suppresser protein p63 in cervical tissues and smears to assess the ability of these proteins to predict progression. Methods: Cervical smears and corresponding tissues were immunostained using mouse monoclonal antibodies against MCM2, MCM5, CDC6, p16 and p63. Smears were treated with a non-ionic surfactant sodium deoxycholate prior to immuno-cytochemistry. The standard ABC method of immunohistochemistry was performed using DAB as the chromogen. The immunostained samples were scored on a 0-3+ scale and staining patterns of smears were compared with those of tissue sections. Sensitivity and specificity for each of these markers were calculated taking histopathology as the gold standard. Result: All the markers were positive in malignant and dysplastic cells. MCM protein expression was found to be up-regulated in LSIL, HSIL and in malignancies to a greater extent than p16 as well as p63. CDC6 protein was preferentially expressed in high grade lesions and in invasive squamous cell carcinomas. A progressive increase in the expression of DNA replication licensing proteins in accordance with the grades of cervical intraepithelial lesion suggests these markers as significant to predict malignant potential of low grade lesions in cervical smears. Conclusion: MCMs and CDC6 can be applied as biomarkers to predict malignant potential of low grade lesions identified in screening programmes and retesting / follow up might be confined to those with high risk lesions alone so that overuse of resources can be safely avoided.

Cancer notification in India.

In many developed countries, notification of cancer cases is compulsory. Developing countries including India accounts for more than half of new cancer cases in the world, however notification of cancer is not yet mandatory. The primary purpose of notification is to effect prevention and control and better utilization of resources. It is also a valuable source for incidence, prevalence, mortality and morbidity of the disease. Notification of cancer will lead to improved awareness of common etiologic agents, better understanding of common preventable causes and better utilization of health resources with better monitoring and evaluation of the effectiveness of health programs such as cancer screening and cancer treatment programs, which ultimately might improve survival. Notification of cancer can be done by the doctor or the hospital. Akin to the integrated disease surveillance project where more than 90% of the districts report weekly data through E-mail/portal, notification of cancer can be implemented if it is incorporated into the National Program for Prevention and Control of Cancer, Diabetes, Cardiovascular diseases and Stroke scheme. The need of the hour is cancer notification in India.