Clinic-pathological aspect of gastro-intestinal stromal tumors at tertiary care Hospital India / Aspecto clínico e patológico dos tumores estromais gastrointestinais em hospital terciário da Índia

Abstract Background This study defines the disease profile in south Indian population and determine the clinic-pathological aspects of Gastro-Intestinal Stromal Tumors. Method In this prospective study patients diagnosed of gastrointestinal stromal tumors were taken thorough clinical examination and a database of Anthropometric details and clinical details were analyzed. Pathological data included tumor size, presence or absence necrosis, mitotic counts, immunohistochemistry for CD-117, CD-34. Results There were 44 patients with confirmed diagnosis of gastro-intestinal stromal tumor. The highest incidence was found in the 6th decade. The most common symptoms were abdominal pain and gastrointestinal bleed. Stomach was most frequent site for gastro-intestinal stromal tumors. Immunochemistry for CD-117 was positive in 93.18% cases. Majority of tumors (79.5%) had pure spindle cell morphology and mitotic activity showed that 34% of the GISTs were of the high risk group. Forty two patients were suggestive of surgery as the primary treatment after presentation. Conclusion Abdominal pain was the most common presenting complaint. Majority of the tumors aroused from the stomach. The majority of the tumors had pure spindle cell morphology and 93% of the tumors were CD-117 positive. A significant relationship between tumor size, tumor necrosis and mitotic activity with large tumors having necrosis and high mitotic rate having high risk of malignancy, was observed. Surgical resection is considered mainstay of treatment of gastro-intestinal stromal tumor. Imatinib therapy should be given to patients in moderate to severe risk categories.

Resumo Justificativa Este estudo define o perfil da doença na população do sul da Índia e determina os aspectos clínicos e patológicos dos tumores estromais gastrointestinais. Método Neste estudo prospectivo, os pacientes diagnosticados com tumor estromal gastrointestinl foram submetidos a um exame clínico completo, e uma série de dados dos pacientes, incluindo detalhes antropométricos e clínicos, foram analisados. Os dados patológicos incluíram tamanho do tumor, presença ou ausência de necrose, contagem mitótica e imuno-histoquímica para CD-117, CD-34. Resultados Havia 44 pacientes com diagnóstico confirmado de tumor estromal gastrointestinal. A maior incidência foi encontrada na 6ª década de vida. Os sintomas mais comuns foram dor abdominal e sangramento gastrointestinal. O estômago foi o local mais frequente para tumores estromais gastrointestinais. A imuno-histoquímica para CD-117 foi positiva em 93,18% dos casos. A maioria dos tumores (79,5%) apresentava morfologia pura de células fusiformes e a atividade mitótica mostrou que 34% dos GISTs pertenciam ao grupo de alto risco. Quarenta e dois pacientes receberam indicação para cirurgia como tratamento primário após a apresentação. Conclusão A dor abdominal foi a queixa mais comum. A maioria dos tumores afetava o estômago, apresentava morfologia pura de células fusiformes e 93% eram CD-117 positivos. Foi observada uma relação significativa entre o tamanho do tumor, a necrose tumoral e a atividade mitótica, com os tumores grandes apresentando necrose e alta taxa mitótica com alto risco de malignidade. A ressecção cirúrgica é considerada o principal tratamento do tumor estromal gastrointestinal. A terapia com imatinibe deve ser administrada a pacientes em categoria de risco de moderadas a grave.

In vitro cardiogenesis can be initiated in human CD34+ cells.

BACKGROUND: The extensive damage that occurs in the cardiac tissue after myocardial infarct is the major concern in post infarct management. It is very well known that adult stem cells mobilized by administration of G-CSF result in homing of stem cells into the damaged myocardium. This is because of the fact that stem cells have the ability to proliferate and capacity to generate into multiple cell lineages. METHOD: A healthy donor was selected as per the guidelines given by the institutional ethical committee and Helsinki declaration. The donor was given G-CSF 5 microg/kg/day and stem cells were harvested from the peripheral blood using Fresenius ASTec204 cell separator. The PBSC were then evaluated by immunohistochemical staining using anti-human CD34 monoclonal antibodies. The cells were then cultured in DMEM with 10% FCS for 17 weeks and in vitro cardiogenesis was initiated by adding 4 microM/l 5'Azacytidine. RESULTS: In vitro cardiogenesis was initiated in pure CD34+ cells with 5' Azacytidine. The cells showed spontaneous beating after 24 hours of treatment and after 5 weeks, the cells connected with the adjoining cells by a myotube. In these cells, expression of myosin light chain (MLC2v) gene and GATA-4 transcription factor validated the development of cardiomyocytes. CONCLUSION: It is observed that the transplantation of autologous stem cells/fetal cardiomyocytes in the heart scar tissue developed due to infarct, limited the scar expansion, and prevented post infarct heart failures. Homing process due to the transplantation of autologous stem cells is time consuming; therefore, transplantation of cardiomyocytes developed from autologous stem cells could be the future method of correcting the infracted myocardium.

study of angiogenesis in human colorectal tumors by using anti-Cd34 antibody. [Assessment by light microscope and computer-aided image analysis system]

Angiogenesis plays a crucial role in tumorigensis; several reports have described a significant increase in microvessel density [MVD] in colorectal carcinogenesis. There are several methods to measure the angiogenesis in neoplasms, but immunohistochemistry seems to be the mainstay of all. This method enable us to measure the tumor microvessel densities highlighted by using antibodies directed against endothelial cell markers like CD31, CD34 or others; then assessment of MVD by manual count of the number of microvessels in what appears to be the most vascular area of the tumor [called the hot spot] using a protocol described by Weidner et al. Automated cellular imaging system is used to analyze immunohistochemically stained slides studies have shown that the device offers accurate precision and reproducibility of immunostained slide analysis exceeding that possible with manual evaluation which was the prevalent method. To assess the angiogenesis in normal, adenomatous [benign] and malignant colorectal tissues using CD34 and the microvessels will be measured both manually by hot spot method as the MVD and by the use of computerized image analysis system as fraction area, we correlate between microvessels density and fraction area with various clinicopathological parameters in colorectal cancer [CRC], and to compare between the results which obtained from both methods. Paraffin embedded archival materials from 50 cases including three normal resection [non tumoruos] margins, 12 benign colonic lesions and 35 colonic adenocarcinoma were used. 5mm section were cut and they were stained by anti CD34 antibody Angiogenesis was measured as MVD by two methods: manually by light microscope and by a computer image analysis system [as fraction area]. Then the MVD and fraction area were correlated with different clinicopathological parameters. This study demonstrate that there is a statistical difference in MVD and fraction area in both hot spot method and CIAS respectively between benign and malignant tumors. P value < 0.05 in hot spot method and less than 0.001 in CIAS and there was highly significant correlation between MVD and fraction area with the grade. There was significant increase in MVD and fraction area from well differentiated to moderately differentiated and to poorly differentiated. There was no significant correlation between MVD and lymph node involvement by hot spot method but CIAS proved a significant correlation between fraction area and lymph node involvement. Both methods [hot spot and CIAS] proved no significant correlation with age, sex, size of the tumor, site of the tumor, stage of the tumor and the number of lymph node involvement

Study of angiogenesis, P53 and NM23 expression in colorectal carcinoma

Colorectal carcinoma [CRC] is not uncommon in Egypt. Sustained angiogenesis is characteristic of several pathological conditions including tumor growth. Many researches were conducted to investigate the role of P53 in colorectal carcinogenesis; also the role of NM23 in tumor progression and for metastasic potential is not so clear in CRC. In this research we are aiming to study the pattern and density of angiogenesis in colorectal carcinomas, in addition to other histopathological prognostic factors. Besides we are also aiming to study the expression of P53 and NM23 and the relation between these three factors [Angiogenesis, P53 and NM23] in different grades and stages of colorectal carcinomas. The study comprised 44 resection specimens of colorectal carcinomas collected from specimens of department of pathology, Faculty of Medicine, Tanta University Hospital and from same of the private laboratories. Each block was stained by hematoxylin and eosin [H and E], immunohistochemical stain for CD34, VEGF, P53 and NM23. Histopathological assessment of grading was done according to WHO classification and staged according to TNM classification. The present study includes 44 cases of colorectal adenocarcinoma 35 were male patients and 9 cases were females. The age range was 20 to 80 years. The patients were grouped into three groups as follows: Group [1]: CRC without nodal or distant metastasis, group [II]: CRC with nodal but no distant metastasis and group [III]: CRC with distant metastasis. Most of the tumors were conventional invasive adenocarcinomas, on studying angiogenesis we found that the relation of microvascular density [MVD] detected by CD34 was not significant with the tumor size. Besides there were a significant results between the microvascular density and metastatic history of the disease. There were significant results between VEGF expression and the studied variables. The study ol apoptosis using P53 reveals significant relation between it and the studied variables but not with the nodal or blood metastasis, there was an inverse relation between NM23 and both VEGF and P53, but the results was not significant with the tumor grade and size, so in the present study the relation between VEGF, P53 and NM23 was significant but each marker have its own variable result with the grade .size and the stage of the tumor. In the present study it can be concluded that MVD at vascular hot spots is a very important predictive factor in CRC in addition to VEGF .P53 over expression has an impact on the biological behavior of CRC being more expressed in biological aggressive tumors and it has a role in angiogenicStudy of Angiogenesis, P53 and NM23 Expression in Colorectal Carcinoma activity of the tumor. We also concluded that NM23 is an important metastatic suppressor gene in CRC. So NM23, P53, VEGF and MVD by CD34 all are important to predict the outcome of the disease and they can be used as a guide for treatment. MVD, microvascular density; VEGF, vascular endothelial growth factor; CRC, colorectal carcinoma

The possible cost effectiveness of peripheral blood stem cell mobilization with clophosphamide and the late addition of G-CSF

The purpose of this study was to develop a cost-effective protocol for the mobilization of peripheral blood stem cells (PBSC) in patients with malignancy. Thirty consecutive patients were randomized to mobilize PBSC with the late addition of a standard 250 microg dose of G-CSF (Neutrogen) from day 8 or early addition of the same dose of G-CSF from day 2, following cyclophosphamide (CY) 4 g/m2. The median yield of CD34+ cells from evaluated patients was 7.87 x 10(6)/kg (range, 2.06-27.25), collected in a median of four apheresis (range, 2-9). Target CD34 + cell doses > or = 2.0 x 10(6)/kg were achieved in all patients able to be evaluated. There were no statistically significant differences in CD34+ cell yields or toxicities. Overall engraftment occurred with median days to neutrophils > or = 0.5 x 10(9)/L or platelets > 20 x 10(9)/L of 11 and 17 days, respectively. However, the duration of G-CSF administration was markedly shorter in the late use of G-CSF group than in the early use of G-CSF group, with a median of 9 days compared with 15 days (p>0.001). PBSC harvesting after priming with CY plus delayed use of G-CSF made it a safe and cost-effective procedure.

Comparison of CD34+ subsets and clonogenicity in human bone marrow, granulocyte colony-stimulating factor-mobilized peripheral blood, and cord blood

To compare the clonogenicity and distribution of CD34+ subsets in bone marrow (BM), granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood (PB) and cord blood (CB), we analyzed in vitro colony formation and CD34+ cells co-expressing differentiation molecules (CD38, HLA-DR), myeloid associated molecules (CD13, CD33), a T-cell associated molecule (CD3), and a B-cell associated molecule (CD19) from mononuclear cells (MNCs) in the three compartments. The proportions of CD34+CD38- cells (BM: 4.4+/-2.8%, PB: 5.3+/-2.1%, CB: 5.9+/-3.9%) and CD34+HLA-DR cells (BM: 4.7+/-3.4%, PB: 5.5+/-2.3%, CB: 6.1+/-3.7%) did not differ significantly among the compartments. In contrast, a significantly higher proportion of CD34 cells of PB and CB co-expressed CD13 (75.0+/-11.4%, 77.7+/-17.3%) and CD33 (67.1 +/-5.7%, 56.8+/-10.3%) compared with those of BM (43.0+/-6.3%, 27.6+/-5.1%) and a significantly higher number of granulocyte-macrophage colony-forming units (CFU-GM) and erythroid burst-forming units (BFU-E) were detected in MNCs derived from PB and CB compared with those from BM (p<0.01). The proportion of CD34+CD19+ cells was higher in BM (34.9+/-11.9%) than those in PB (5.6+/-3.0%) and CB (4.7=2.1%) (p<0.05). The proportion of CD34+CD3+ was comparable in all three compartments. In conclusion, our findings show that MNCs of mobilized PB and CB display similar phenotypic profiles of CD34+ subsets and clonogenicity, different from those of BM.