Siriraj Medical Journal

1-5

Authors: Tumtip Sangruchi, M.D., Panitta Sithinamsuwan, M.D., Jane Manonukul, M.D., Suteekanit Hahtapornsawan, M.D., Nattawut Sermsathanasawadi, M.D., Methee Chayakulkeeree, M.D., Theerapong Krajaejun, M.D., Ankana Chaiprasert, Dr.rer.nat, Wanchai Wanachiwanawin, M.D.

Objective: Arterial pythiosis, caused by Pythium insidiosum, is a life threatening condition. The lesions usually involve main arteries of the lower extremities, which extend to the abdominal aorta. Early amputation is required to save patient’s life. The adequacy of amputation needs to ensure that the resection margins are devoid of the organisms. The aim of this study is to suggest the level that should be devoid of organisms for surgical amputation. Methods: We studied amputated limbs of patients diagnosed as arterial pythiosis. The major arteries of the limbs were dis- sected, then serial cross cut at 0.3-0.5 cm intervals, and consecutively submitted from proximal to distal segments for routine tissue processing and staining. Segments of non-in amed vessels continuing above or below the level of arterial occlusion were identi ed. To demonstrate fungal-like hyphae, all corresponding blocks of non-in amed arteries were stained with Grocott’s methenamine silver staining (GMS) andthoroughly examined under a microscope. Results: From June 2010 to June 2011, there were 4 cases of arterial pythiosis. Chronic arteritis obliterans, acute necrotizing arteritis with thrombosis, or ruptured aneurysm was demonstrated from distal to proximal segments without skip lesions. At the level of 5 cm. from the occlusion, neither arteritis nor fungal-like hyphae was identi ed. Conclusion: Based on the evidence from only 4 cases, we have proposed that at 5 cm. or more above the occlusion, the non- in amed arteries were free of P. insidiosum, while fungal hyphae might be observed in in amed soft tissue.

9-11

Authors: Jaruda Kobkitjaroen, M.D., Jintana Jaiyen, M.Sc., Siriporn Kongkriengdach, B.Sc., Sumalee Potprasart, B.Ed., Ronnachai Viriyataveekul, M.D.

The Crithidia luciliae immuno uorescence test (CLIFT) is widely used in clinical laboratories for anti-double stranded DNA (anti-dsDNA) detection. It has comparable sensitivity and speci city to the standard method, Farr assay, but employs simpler and safer techniques. A number of CLIFT kits are commercially available. We evaluated the sensitivities and speci cities of three commercial CLIFT kits, including IMTEC, Hemagen and Euroimmun. The IMTEC kit detected IgG, IgM and IgA classes, while the Hemagen and Euroimmun kits detected only the IgG class. Farr assays were performed when three kits gave discrepant results. The gold standard methods were the consensus results of all three kits together with the Farr assay. Out of one-hundred and thirty sera, 111 sera (85%) gave concordant results by all three CLIFT kits and 19 sera (15%) gave discrepant results. Sensitivities and speci cities of the IMTEC, Hemagen and Euroimmun kits were 97.4% and 94.5%, 66.7% and 100%, and 89.7% and 94.5%, respectively. The size of the IMTEC C. luciliae was smaller than those of the other two kits, leading to more dif culty in slide reading. Factors affecting the kit’s performance may include substrate preparation and immunoglobulin class detection.

16-20

Authors: Jantima Roongruangchai, D.D.S., Ph.D., Sataporn Menbangphung, M.S., Kasem Kuedpuch, B.Sc., Nachaya Amonmettajit, B.Sc., Pae Pochanasomboon, B.Sc., Kosol Roongruangchai, M.S., M.D.

Background: Esophageal atresia (EA) or stenosis is well known among the congenital abnormalities of the esophagus which may result from the failure or incomplete recanalization after solid stage during the 8th week of human development or deviation of the tracheo-esophageal septum. The occlusion and recanalization theory of gut development cannot explain some types of esophageal atresia. Objective: To study the serial section of continuous stages of esophageal development of chick embryos from related human Carnegie stages 14-23, observe histologically whether there are occlusion and recanalization processes, at which stages and to calculate the mitotic and apoptotic indices of the epithelial lining of the esophageal lumen as well as measuring the lumen areas of each stage of esophageal development. Methods: The fertilized Leghorn hen eggs (Gallus domesticus) were incubated at different time intervals to gain ve em- bryos for each developing stage 14-23. They were processed histologically into serial sections after the total bodies were photographed. The serial sections were observed at the esophageal area by one skipped ve sections technique. The length and lumen area were measured, total epithelial cell count, mitotic and apoptotic cell counts were performed and the mitotic and apoptotic indices were calculated. Results: The lengths of the esophagus increased with advanced age from 126.80 ± 3.03 μm at stages 14 to 7, 234.80 ± 396.62 μm at stage 23. The lumen areas decreased from stages 14-16 and increased from stages 17-23. There was no evidence of occlusion at any stage. At the stages of increasing the lumen area there was also no evidence of vacuole formation. The mitotic indices decreased from stages 15-20 and were constant from stage 21-23, which showed no correlation with the lumen area. The apoptotic index was highest at stage 16 which also showed no correlation with the process of recanalization. Conclusion: By studying in the continuous stages of esophageal development, we could not demonstrate histologically the occuluded esophageal lumens, or vacuolation of the lumens at any stage. In the same way, the mitotic and apoptotic indices show no correlation with the lumen area. These results lead to the conclusion that processes of occlusion and recanalization never occur in the normal esophageal development.

24-29

Authors: Chaicharoen Tantanate, M.D.

The bleeding time is one of the oldest tests for primary hemostasis which is still performed nowadays.This test evaluates mainly the primary hemostatic function. Some preanalytical variables including patients’ physiology and hema- tological status may impact test results. Various protocols of bleeding time are performed. Each method has pros and cons, although the template bleeding time is currently preferred over other methods because it is well-standardized. Interpretation of the results according to a well-established reference range is important. At the moment, indications of bleeding time are very limited due to its poor sensitivity and reproducibility. More sensitive and speci c assays have been developed for the diagnosis of speci c conditions. However bleeding time is still useful in uremic patients. A cautionary note - this test should not be used as a screening test for prediction of bleeding preoperatively.