Changing scenario of therapeutic apheresis in India in the last 14 years.

There has been a remarkable change in the scenario of therapeutic apheresis in the last 14 years in India. The crude method of manual removal of blood followed by separation of plasma by gravity, keeping it in the bottle for a long time, has now been totally replaced by plasmapheresis, centrifugation, membrane filtration, and immunoadsorption techniques. The indications for use have also changed from a list of limited indications in the beginning to include all immune complex disorders. The clinical beneficiaries have also increased from blood bankers to nephrologists and immunologists in addition to oncologists. Efforts are now underway with the help of the Indian Society for Apheresis (founded in 1985) to popularize the newer methods of cryofiltration, photopheresis and heparin extracorporeal low-density lipoprotein (HELP) and DALI apheresis systems besides the specialized techniques of immunoadsorption using filters, columns, or ligands. This is suggestive of a positive trend for the treatment of immune complex disorders.     

Parental Communication Not to Smoke and Adolescent Cigarette Smokers' Readiness to Quit: Differences by Age

There is limited information on the relationship between parental practices that specifically discourage current cigarette smoking and adolescent cessation, and how this relationship varies by age. Among 1629 adolescent smokers, self-reported receipt of parental communication not to smoke was significantly and positively associated with readiness to quit. The strength and significance of this association decreased from early to middle adolescence and was not significant in late adolescence.     

Double skin island pectoralis major myocutaneous flap with nipple-areola complex preservation: a case report.

A horizontal double skin island pectoralis major myocutaneous flap with preservation of nipple-areola complex is described, a different modification used to reconstruct a complex defect following radical resection of a locally advanced gingival carcinoma. The skin islands are horizontally separated by a distance of 2.5 cm but rely on one vascular pedicle for their blood supply. The muscle between the skin islands provides the soft tissue bulk between the internal mucosal and external skin lining. The need for a separate segmental flap or a split-thickness skin graft for mucosal lining was eliminated.     

EGFR gene amplification in breast cancer: correlation with epidermal growth factor receptor mRNA and protein expression and HER-2 status and absence of EGFR-activating mutations.

The human epidermal growth factor receptor (HER) family of receptor tyrosine kinase has been extensively studied in breast cancer; however, systematic studies of EGFR gene amplification and protein overexpression in breast carcinoma are lacking. We studied EGFR gene amplification by chromogenic in situ hybridization (CISH) and protein expression by immunohistochemistry in 175 breast carcinomas, using tissue microarrays. Tumors with >5 EGFR gene copies per nucleus were interpreted as positive for gene amplification. Protein overexpression was scored according to standardized criteria originally developed for HER-2. EGFR mRNA levels, as measured by Affymetrix U133 Gene Chip microarray hybridization, were available in 63 of these tumors. HER-2 gene amplification by fluorescence in situ hybridization (FISH) and protein overexpression by immunohistochemistry were also studied. EGFR gene amplification (copy number range: 7-18; median: 12) was detected in 11/175 (6%) tumors, and protein overexpression was found in 13/175 (7%) tumors. Of the 11 tumors, 10 (91%) with gene amplification also showed EGFR protein overexpression (2+ or 3+ by immunohistochemistry). The EGFR mRNA level, based on Affymetrix U133 chip hybridization data, was increased relative to other breast cancer samples in three of the five tumors showing gene amplification. Exons 19 and 21 of EGFR, the sites of hotspot mutations in lung adenocarcinomas, were screened in the 11 EGFR-amplified tumors but no mutations were found. Three of these 11 tumors also showed HER-2 overexpression and gene amplification. Approximately 6% of breast carcinomas show EGFR amplification with EGFR protein overexpression and may be candidates for trials of EGFR-targeted antibodies or small inhibitory molecules.     

Feasibility of using tissue microarrays for the assessment of HER-2 gene amplification by fluorescence in situ hybridization in breast carcinoma.

Tissue microarrays (TMAs) have been commonly used to study protein expression by immunohistochemistry (IHC). However, limited data exist on the validity of using TMAs to study gene amplification. In this study, we evaluated the feasibility of using breast carcinoma TMAs to study HER-2 gene amplification by fluorescence in situ hybridization (FISH). In addition, hormonal receptor status (ER and PR) and HER-2 protein overexpression by IHC were also studied, and results were compared with whole tissue sections. FISH for HER-2 was performed on formalin-fixed paraffin-embedded tissue from 114 invasive breast carcinomas both on whole tissue sections and on TMAs containing the same tumors. The TMA was created using 0.6-mm tissue cores with four sampled cores per tumor from the same tissue block used for whole section FISH. The PathVysion HER-2 probe kit was used for the FISH analysis. A ratio of HER-2:Chromosome17 > or =2.0 was interpreted as positive for gene amplification. The ER or PR was interpreted as positive when nuclear staining was detected in more than 10% of tumor cells. The HER-2 IHC (HercepTest; DAKO Corp, Carpinteria, CA) results were interpreted as 0, 1+, 2+, and 3+ according to standard criteria. The FISH results in the TMA and whole sections were concordant in 99 out of 101 successfully analyzed cases (99%). The FISH scores were consistent among the two to four cores in the majority of the cases. ER and PR results were concordant between whole sections and TMA cores in 97% (107/110) and 89% (97/109) cases, respectively. The overall concordance for HER-2 status by IHC between whole sections and TMA cores was 86% (94 out of 109 cases). TMAs are a reliable approach to study HER-2 gene amplification in a high throughput manner.     

Chromogenic in situ hybridization for the detection of HER-2/neu gene amplification in breast cancer with an emphasis on tumors with borderline and low-level amplification: does it measure up to fluorescence in situ hybridization?

We compared chromogenic in situ hybridization (CISH) with fluorescence in situ hybridization (FISH) for assessing HER-2/neu gene amplification using tissue microarrays (TMAs) made from formalin-fixed, paraffin-embedded tissue blocks from 113 cases of invasive breast carcinoma. TMAs were created using 0.6-mm tissue cores with 4 sampled cores per tumor. For both assays, a HER-2/chromosome 17 signal ratio of 2.0 or more was considered positive for gene amplification. The average ratio of cores from the same tumor was used for determination of gene amplification status of that particular tumor Of 113 cases, 102 were tested successfully by both assays. The results were concordant in 100.0% of cases (63 amplified; 39 nonamplified). All 22 cases of borderline (ratio, 2.0-2.5) or low-level (ratio, 2.6-3.9) amplification by FISH also showed HER-2 gene amplification by CISH. CISH is as sensitive as FISH in detecting borderline and low-level HER-2 amplification. Reliable recognition of the invasive carcinoma area by light microscopy and preservation of the test slides are added advantages of CISH. CISH performs as well as FISH in the analysis of HER-2 gene amplification in breast cancer and might have advantages in certain situations.