Evaluation of Zymodemes of Pathogen and Nonpathogen Entamoeba histolytica

Entamoeba histolytica is the protozoan responsible for human amoebosis. It infects 10% of the world population, causing 50 million cases of dysentery or liver abscesses and to death of 100.000 humans per year. As with other parasitic infections, amoebosis is a consequence of poverty and bad sanitary conditions. According to epidemiological studies, only 10% of the effected people are infected with pathogenic strains. By enzyme electrophoresis, 23 zymodemes of E. histolytica have been identified. Of these, nine have been associated with pathology in the host. In this study, seven E. histolytica isolates were analized for their zymodeme patterns and the enzymes used to classify E. histolytica include glucosphosphate isomerase (GPI), hexokinase (HK), L-Malate (ME), and phosphoglucomutase (PGM) by observing the migration pattern of isoenzyme bands in polyacrylamide gel electrophoresis (PAGE). Our isolates were taken from different geographical parts; Diyarbakır (four isolates), Izmir (two isolates) and Ankara (one isolate) regions. HK enzyme is characterized by two fast-migrating bands in four isolates of symptomatic patients (three isolates from Diyarbakır region, and one isolate from Izmir region) and one isolate from on asymptomatic patient. PGM band were observed in three isolates (Diyarbakır region). GPI band was observedas in two isolates (Diyarbak›r region), whereas as a beta band in one isolate (Diyarbakır region). ME band is not found in any of the isolates

Ankara'da i?nsan ve buza?ılarda Cryptosporidium spp. Varlı?ının nested PCR ve carbol fuchsin boyama yöntemleri ile belirlenmesi]

In this study, a total of 130 fecal samples, obtained from 98 humans and 32 calves from different hospitals and dairy farms in Ankara, respectively were analyzed with carbol fuchsin staining and nested PCR to determine the presence of Cryptosporidium spp. Nested PCR revealed a total of 13 (10.0%) positive amplification results; 12 (37.5%) were from calves and 1(1.02%) from humans. Oocysts were observed in 8 (6.15%) fecal samples belonging to 7 (21.88%) calves and 1 (1.02%) human by carbol fuchsin staining. Concerning the detection technics, it was concluded that nested PCR can be successfully used in diagnosis of cryptosporidiosis, and although carbol fuchsin staining method has sufficient specificity it is less sensitive, especially in cases characterized with excretion of fewer oocysts

In vitro efficacy of hyperbaric oxygen therapy against Leishmania tropica promastigotes and amastigotes

Aim: To assess the efficiency of hyperbaric oxygen (HBO) therapy on L. tropica, which is the major causative agent of cutaneous leishmaniasis in Turkey. Materials and methods: the effects of HBO on Leishmania tropica promastigote and amastigote forms were studied at a pressure of 2.5 atmosphere absolute (ata). We examined the cytotoxic effect of HBO on promastigotes using the trypan blue exclusion test and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye-reduction assay. To investigate the effects of HBO treatment on the growth of the promastigotes, HBO-treated and non-HBO-treated cell culture flasks were incubated at 26 °C for 3 days, while the effects of HBO on L. tropica amastigotes were detected using the acridine orange staining method. Results: We observed that the cytotoxic eff ect emerged aft er 2 h and reached a maximum at 6 h with both methods. the number of live promastigotes was monitored for 24 h after treatment and there was a slight elevation at 2 h of treatment, but significant suppression was seen after 4 and 6 h of treatment (P ;lt; 0.001). We detected that the effect of HBO on L. tropica amastigote forms increased by 3.3%, 17.2%, and 38.5% at 2, 4, and 6 h, respectively. Conclusion: the results presented here indicate that HBO treatment could be used as supportive therapy in association with chemotherapeutic agents against L. tropica infection.Amaç: Bu çalışmanın amacı, Türkiye’de kutanöz leishmaniasisin en önemli etkeni L. tropica üzerinde HBO tedavisinin etkinliğini değerlendirmektir. Yöntem ve gereç: Leishmania tropica promastigot ve amastigot formları üzerinde hiperbarik oksijenin etkileri 2,5 atmosfer basıncı (ata) altında çalışıldı. HBO’nun promastigot formları üzerindeki sitotoksik etkileri, tripan mavisi testi ve MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) yöntemiyle incelendi. HBO tedavisinin L. tropica promastigot formları üzerinde etkilerini araştırmak amacıyla HBO uygulamasına maruz bırakılmış ve bırakılmamış hücre kültür plakları 26oC’de üç gün boyunca inkübe edildi. Diğer taraftan HBO’nun L. tropica amastigot formları üzerine olan etkileri akridin oranj boyama yöntemiyle tespit edildi. Bulgular: Her iki yöntem ile sitotoksik etkinin iki saat sonra başladığı ve altı saat sonra en üst seviyeye ulaştığı gözlemlendi. Canlı promastigotların sayısı tedaviden sonra 24 saat boyunca izlendi ve tedavinin ikinci saatinin sonunda promastigot sayılarında hafif bir artış olduğu, dört ve altı saat sonrasında ise promastigot sayılarının artışında önemli bir azalma olduğu tespit edildi (P 0,001). Diğer taraft an HBO tedavisinin L. tropica amastigot formları üzerinde etkisinin 2, 4 ve 6. saatler için sırasıyla % 3,3, % 17,2 ve % 38,5 olduğu gözlemlendi. Sonuç: Bu çalışmada elde edilen sonuçlar, HBO tedavisinin L. tropica formlarına karşı kemoterapi ile birlikte destekleyici tedavi olarak kullanılabileceğini göstermektedir

Applications of sterile larvae of Lucilia sericata for the treatment of chronic wounds

Bu çalışmada, maggot tedavisinin (MT) etkinliğini değerlendirmek ve en uygun ve etkin orijinal steril larva içeren paket dizaynlarının geliştirmek amaçlanmıştır. Maggot tedavisinin en iyi kullanım ve etkinliği için larva içeren paket sistemi geliştirilmiştir. Çalışmada çeşitli boyutlarda (ortalama 11,9±11,8 cm2) yaralar bulunan 23 diyabetik ayak ülseri (DAÜ), iki iskemik kronik yaralı olmak üzere toplam 25 kronik yaralı hasta alınmıştır. Yaranın klinik durumu tedavi öncesi ve sonrası ayrı ayrı değerlendirilmiştir. Tedavi öncesi, yaralarda Gram pozitif kok ve Gram negatif basil en sık olarak saptanmıştır. Hastalara MT (ortalama 16,2±8,6 gün) steril ikinci dönem larvaları içeren paket (ortalama 3,6±1,6 paket) ile birlikte hiperbarik oksijen (HBO) tedavisi uygulanmıştır. Tedavi sonucunda 19 hastanın (%76) yarasında tam bir debridman sağlanırken, altı hastada (%24) kısmi bir debridman oluşmuştur. Sonuç olarak; kronik yarası olan hastaların tedavilerinde steril maggot tedavisinin “paket” olarak uygulanmasının pratik olması yanında tedavide etkin olabildiği gözlenmiştir.The objective of this study was to assess the efficacy of maggot therapy and to develop decontained maggot designs for the best use and effect. In this study, we could maintenance of Lucilia sericata colony and sterile maggots in the insectariums. Contained maggot designs would be developed for the best use and effect for maggot therapy. We have enrolled 25 patients with non-healing chronic wounds (23 diabetic foot wounds, 2 ischemic foot wounds,) various wound size (mean 11,9±11,8 cm2 ). The clinical state of the wounds documented before and after maggot therapy. Gram-positive cocci and Gram-negative bacilli were the most frequently isolated from patients with chronic wounds in pretreatment. Along with packet that contains the sterile larvae of the second stage (average mean number of applications 3,6±1,6 packet; average mean 16,2±8,6 days) hyperbaric oxygen (HBO) was applied to patients. As a result of treatment of 19 patients (76%) provided a complete wound debridement, on the other hand, a partial debridement was occurred in six patients (24%). As a result, application of maggot therapy as a "package" in the treatment of patients with chronic wounds is practical and effective