Spor Oluşturan Bağırsak Parazitlerinin Pentaplex Real-Time PCR ile Tanımlanması

Spore-forming intestinal parasites; Cryptosporidium spp., Cyclospora spp., Cytoisospora spp., Encephalitozoon spp., and Sarcoystis spp. are very common in immunocompromised patients, but these parasites are overlooked by healthcare proffesionals. It was aimed to develop a new pentaplex real-time PCR panel for the identification of spore-forming intestinal parasites in this study. Methods: Primer-probes for pentaplex real-time PCR were designed using the “PrimerQuest Tool (Integrated DNA technologies, Coralville, USA) software program” and “Multiple sequence alignment use a computer software Primer Express™Software v3.0.1 Lience (ThermoFisher Scientific, Waltham, USA)”. The primer-probes designed in the study were spore-forming Cryptosporidium spp. (ATCC®87715™), Cyclospora spp. (ATCC®PRA-3000SD™), Cystoispora spp. (KF648871), Encephalitozoon spp. (FJ026010) and Sarcocystis spp. (ATCC®CCL-70) parasites were amplified with DNA isolates from the American Type Culture Collection (ATCC) and then these primer-probes were validated with 232 DNA samples obtained from the stools of the patient samples. Results: It was found that Cycle Threshold (Ct) ±25.7, Standard curve (R2 ): ±0,993, and Efficiency (E): %96,1 according to the results of multiplex real-time PCR analysis. Similar results were found in pentaplex real-time PCR analysis of DNA isolates of stool samples. When the pentaplex real-time PCR results of DNA samples isolates from stool samples were compared with the positivie predictive value results of traditional methods, it was found that the pentaplex results were higher. Conclusion: The new designed pentaplex real-time PCR panel can be used in the diagnosis of spore-forming intestinal parasites, which are very common in immunocompromised patients. Thus, the diagnosis of five different parasites can be made faster, more economically and faster with a single reaction.Spor oluşturan bağırsak parazitleri; Cryptosporidium spp., Cyclospora spp., Cytoisospora spp., Encephalitozoon spp., ve Sarcoystis spp. bağışıklığı baskılanmış hastalarda çok yaygındır, ancak bu parazitler sağlık personeli tarafından göz ardı edilir. Bu çalışmada spor oluşturan bağırsak parazitlerinin tanımlanması için yeni bir pentaplex real-time PCR panelinin geliştirilmesi amaçlanmıştır. Yöntemler: Pentaplex real-time PCR için primer-problar “Primer Quest Tool (Integrated DNA technologies, Coralville, USA) software programı” ve “Multiple sequence alignment use a computer software Primer Express™Software v3.0.1 Lience (ThermoFisher Scientific, Waltham, USA)” programı kullanılarak tasarlandı. Çalışmada tasarlanan primer-problar, Amerikan Tipi Kültür Koleksiyonu (ATCC)’nunda temin edilen, spor oluşturan Cryptosporidium spp. (ATCC®87715™), Cyclospora spp. (ATCC®PRA-3000SD™), Cystoispora spp. (KF648871), Encephalitozoon spp. (FJ026010), and Sarcocystis spp. (ATCC®CCL-70) parazitlerinin DNA izolatları ile amplifiye edildikten sonra hasta örneklerinden elde edilen 232 DNA örneği ile doğrulandı. Bulgular: Pentaplex real-time PCR analizi sonuçlarına göre analizin döngü Eşiği (Ct) ±25,7, standart eğrisi (R2 ): ±0,993 ve verimliliği (E): %96,1 olarak bulundu. Dışkı örneklerinin DNA izolatlarının pentaplex real-time PCR analizinde de benzer sonuçlar bulundu. Dışkı örneklerinden izole edilen DNA örneklerinin pentaplex sonuçları ile geleneksel yöntemlerin pozitif öngörme değerinin yüksek olduğu bulundu. Sonuç: Bağışıklık sistemi baskılanmış hastalarda çok sık görülen sporlu bağırsak parazitlerinin tanısında yeni tasarlanan pentaplex real-time PCR paneli kullanılabilir. Böylece beş farklı parazitin tanısı tek bir reaksiyon ile daha hızlı, daha ekonomik ve daha hızlı konulabilir

İNSANLARDA HASTALIK YAPAN DİROFİLARİA TÜRLERİ VE DİROFİLARİASİS

Öz Dirofilariasis insanlara rastlantısal olarak bulaşan bir zoonoz hastalıktır. Bu hastalığa doğal olarak birçok evcil ve vahşi hayvana zarar veren Dirofilaria cinsinin filarial nematodları neden olur. Dirofilaria Türkiye’nin en önemli vektörleri olan Culex, Anopheles, Aedes gibi çeşitli sivrisinek türleri tarafından insana bulaşır. Dirofilariasis etkeni Dirofilaria paraziti membran filtrasyon-asit fosfataz histokimyasal boyama yöntemi, serolojik yöntemler ve moleküler yöntemler ile tanımlanabilmektedir. Dirofilaria karnivor hayvanlarda özellikle köpeklerde ve insanlarda ciddi patolojik bozukluklara hatta ölümlere neden olabilen bir parazittir. Son yıllarda, evlerinde köpek veya kedi bakmanın artması nedeniyle insanlara Dirofilariasis hastalığının bulaşma riski giderek artmaktadır. Bu yüzden ülkemizde ve dünyada hastalığın yayılmasını önlemek için kontrol programları uygulanmalıdır. Bu derleme insan Dirofilariasis etkenlerini,  Dirofilariasis’in klinik muayenesini ve laboratuvar tanı yöntemlerini içermektedir.Abstract Dirofilariasis is one of the zoonotic fiilarial infections inadvertently affecting the humans. It is caused by filarial nematodes of genus Dirofilaria, which naturally infects several domestic and wild animals. Dirofilaria is transmitted to human by mosquitoes (Diptera, Culicidae) of various species, such as Culex, Anopheles, Aedes which are probably the most important vectors in Turkey. The Dirofilaria parasite which cause of Dirofilariasis disease can be diagnosed using membrane filtration-acide phosphates histochemical staining, serological methods and molecular methods. Dirofilaria is a parasite that can cause serious pathological disorders and even deaths in carnivorous animals especially dogs and humans. In recent years, people are increasingly at risk of being infected with Dirofilariasis diseases due to increased dog or cat feding in their homes. Therefore, control programmes should be implemented to prevent the spread of the disease in our country and world. This review includes human Dirofilariasis agents, clinical examination and laboratory diagnostic methods of Dirofilariasis

Kutanöz leyişmanyozlu hastalarda etken türlerin PCR-RFLP yöntemi ile tanımlanması

TEZ6657Tez (Yüksek Lisans) -- Çukurova Üniversitesi, Adana, 2008.Kaynakça (s.50-54) var.ix, 61 s. : rnk.res. ; 29 cm.The group of disease the leishmaniasis is caused by obligate intracellular protozoa of genus Leishmania. These are present in 3 different forms: Visceral Lesihmaniasis ( VL), Cutaneous Leishmaniasis (CL) and Mucocutaneous Leishmaniasis (MCL). The Cutaneous Leishmaniasis is skin disease transmitted by infected sand fly. Cutaneous leishmaniasis (CL) is most frequently diagnosed by clinical evaluation and is typical acute lesions can be confused with other dermatological problems, such as sporotrichosis. The Cutaneous Leishmaniosis caused by Leishmania tropica and Leishmania major is a health problem in Çukurova region. Serological tests, Indirect Immunofluorescence Antibody Test (IFAT), the Enzyme-linked Immunosorbent Assay (ELISA), direct microscopic examination of smears and culture are use conventional methods for diagnosis of Cutaneous Leishmaniosis. None of these techniques isn't a rapid and definite diagnosis metods for CL. Furtermore, these techniques are not adequate for species identification due to the morfological similarity of different Leishmania species.Leyişmanyoz, Leishmania cinsi zorunlu hücre içi protozoonların neden olduğu bir grup hastalıktır. Bunların üç faklı şekli vardır: Viseral Leyişmanyoz, Kutanöz Leyişmanyoz ve mukokutanöz leyişmanyozdur. Kutanöz leyişmanyoz enfekte tatarcık sineği ile bulaşan bir deri hastalığıdır. Kutanöz Leyişmanyoz tanısı sıklıkla klinik değerler ile konur ve tipik akut lezyonlar sporotrikoz gibi diğer dermatolojik problemler ile karışabilir. Kutanöz leyişmanyoz Çukurova bölgesinde Leishmania tropica ve Leishmania major'un neden olduğu önemli bir sağlık problemidir. Serolojik testler, indirekt immunflorasan antikor testi (IFAT), enzim-sıvı immun sorbent assay (ELISA), direk mikroskop tanısı ve kültür Kutanöz Leyişmanyoz tanısı için kullanılan geleneksel yöntemlerdir. Bu teknikler Kutanöz Leyişmanyoz tanısı için kesin ve hızlı değildir. Ayrıca bu teknikler Leishmania türlerinin morfolojik benzerliklerden dolayı tiplendirme yapmak için de uygun değildir. Bu yüzden son zamanlarda çeşitli hastalığın tanımı ve tür tayini yapmak için özellikle Polimeraz zincir reaksiyonu (PCR)'na dayalı yöntemler geliştirilmiştir.Bu çalışma Ç.Ü. Bilimsel Araştırma Projeleri Birimi Tarafından Desteklenmiştir. Proje No:TF.2996.YL.1

Identification of neutrophil extracellular trap cells in familial mediterranean fever patients.

TEZ11364Tez (Yüksek Lisans) -- Çukurova Üniversitesi, Adana, 2013.Kaynakça (s. 41-47) var.xii, 49 s. : res. (bzs. rnk.), tablo ; 29 cm.Enfeksiyon hastalıklarında mikroorganizmaların ortadan kaldırılmasında nötrofillerin fagositoz ve degranülasyondan sonraki son silahın nötrofil hücre dışı tuzaklarının olduğu bilinmektedir. Ancak bu hücrelerin otoinflamatuvar hastalıklarda özellikle Ailesel Akdeniz Ateşi hastalığındaki rolü bilinmemektedir. Bu nedenle çalışmada Ailesel Akdeniz Ateşi hastalığı ile nötrofil hücre dışı tuzakları arasındaki ilişki araştırılmıştır. Çukurova Üniversitesi Tıp Fakültesi Çocuk Romotoloji Bilim Dalı’na başvuran Ailesel Akdeniz Ateşi tanısı almış 12 ataklı, 18’i remisyon döneminde ve 12 adet sağlıklı kontrol grubu olmak üzere toplam 42 hastadan kan örneği alındı. Kan örneklerinden nötrofil izolasyonu yapıldı. Nötrofillerdeki nötrofil hücre dışı tuzak hücrelerinin DNA’ları sytox boyası ile, nötrofil elastaz granülleri anti-neutrophil elastase antikoru kullanılarak, histon 3 proteinleri ise anti-histone H3 antikorları kullanılarak mikroskopta görünür hale getirildi. Ataklı olan Ailesel Akdeniz Ateşi hastalarında DNA sayısı 26x106, nötrofil elastaz sayısı 13x106, histon 3 sayısı ise 9x106 olarak bulundu. Ataksız hastalarda DNA sayısı 2x106, nötrofil elastaz sayısı 2x106, 1x106 olarak bulundu. Ataklı ve atak sonrası hastalardaki nötrofil hücre dışı tuzakları arasında anlamlı fark (p?0,05) bulunurken, remisyon dönemindeki ve sağlıklı kontrol grubu arasında anlamlı fark (p>0,05) bulunmadı. Çalışmamızın sonucunda da nötrofillern savunma stratejilerinden biri olan NETOsis olayının ve nötrofil hücre dışı tuzaklarının ataklı hastalarda yüksek derecede artığı remisyondaki hasta grubunda ise herhangi bir yükseliş olmadığı tespit edilmiştir. Bu durum nötrofil hücre dışı tuzaklarının hastalığın prognozunu etkilediğini göstermektedir.In the elimination of microorganisms in infectious diseases, neutrophils are known to be the last weapon of neutrophil extracellular traps after phagocytosis and degranulation. However, the role of these cells in autoinflammatory diseases, especially Familial Mediterranean Fever, is unknown. Therefore, the relationship between familial Mediterranean fever disease and neutrophil extracellular traps was investigated in this study. A total of 42 patients, 12 of whom were diagnosed as Familial Mediterranean Fever who were admitted to Çukurova University, Faculty of Medicine, Department of Children's Romotology, were included in the study. Neutrophil isolation was performed by using histopaque method. The DNA of neutrophil extracellular trap cells in the neutrophils was visualized by sytox staining using neutrophil elastase granules using anti-neutrophil elastase reagent, and histone 3 proteins were exposed by microscopy using anti-histone H3 antibodies.The number of DNA in the patients with Familial Mediterranean Fever was 26x106, the number of neutrophil elastase was 13x106 and the histone 3 number was 9x106. The number of DNA was 2x106, the number of neutrophil elastase was 2x106 and the histone 3 number was 1x106. There was a significant difference (p ,0.05) between the neutrophil extracellular traps in patients after attack and attack, while there was no significant difference between the remission period and healthy control group (p> 0.05). As a result of our study, it was determined that NETOsis and neutrophil extracellular trap cell numbers, which are one of the neutrophil defense strategies, increased in patients with remission and no increase in remission patients. This shows that the number of neutrophil extracellular trap cells affects the prognosis of the disease.Bu çalışma Ç.Ü. Bilimsel Araştırma Projeleri Birimi tarafından desteklenmiştir. Proje No: FYL-2018-10375

Diagnosis of Leishmania species which has Laishmaniasis effect is very.

TEZ9700Tez (Doktora) -- Çukurova Üniversitesi, Adana, 2012.Kaynakça (s. 46-51) var.viii, 52 s. : res. (bzs. rnk.), tablo ; 29 cm.Leyişmanyoz etkeni olan Leishmania türlerinin tanımlanması hastalığın klinik özelliklerini belirlemek, hastalığın seyrini takip etmek ve hastalara uygun tedavi uygulamak için çok önemlidir. Leyişmanyoz’un epidemiyoloji ile ilgili klasik bilgilerde Leishmania tropica (L. tropica) ve Leishmania major (L. major)’un Kutanöz Leyişmanyoz’a, Leishmania infantum (L. infantum) ve Leishmania donovani (L. donovani)’nin ise Visseral Leyişmanyoz’a neden olduğu bildirilmektedir. Son yıllarda, Leishmania türlerinin genotiplerinin ve patojenitesinin belirlenmesinde polimeraz zincir reaksiyonuna (PCR) dayalı moleküler yöntemler geliştirilmiştir. Bu çalışmalar sonucunda KL hikâyesi olmayan VL olgularında L. tropica, VL hikâyesi olmayan KL olgularında ise L. infantum türleri izole edilmiştir. Bu durum literatürde rapor edilen klasik bilgilerin doğru olmadığı, L. infantum ile L. tropica’nın her iki formdaki Leyişmanyoz’a neden olabileceği ihtimalini ortaya çıkartmıştır. Bu çalışmada Kutanöz Leyişmanyoz’lu olgulardan L. tropica (%49), L. infantum (% 36), L. major (%15), Visseral Leyişmanyoz’lu olgularda L. İnfantum (%56), L. donovani (%31) ve L. tropica (%13) türleri izole edildi. Kutanöz Leyişmanyoz’lu olgulardan izole edilen L. infantum ve L. major’un haplotip, L. tropica’nın heterotip olduğu tespit edildi. Visseral Leyişmanyoz’lu olgulardan izole edilen L. infantum, L. donovani ve L. tropica’nın haplotip olduğu tespit edildi. Kutanöz Leyişmanyoz’lu olgulardan izole edilen L. infantum’lar ile Visseral Leyişmanyoz’lu olgulardan izole edilen L. infantum’ların nükleotit dizileri arasında benzerlik görülmektedir. Bu durum Kutanöz Leyişmanyoz olgularından izole edilen L. infatum türlerinin Visseral Leyişmanyoza neden olabileceğini düşündürmektedir. Çalışmada Kutanöz Leyişmanyoz hikayesi olmayan Visseral Leyişmanyoz olgularında ise L. tropica türü tespit edildi. Çalışmanın sonucunda Visseral Leyişmanyoz etkeni olarak bilinen L. infantum’un Kutanöz Leyişmanyoz’lu olgularda da görülebileceği, Kutanöz Leyişmanyoz etkeni olarak bilinen L. tropica’nın Visseral Leyişmanyoz’lu olgularda da görüldüğü saptandı.Diagnosis of Leishmania species which has Laishmaniasis effect is very important for determination of clinic features of disease, pursuing the course of disease and apply proper therapy to patients. In the classical information about epidemiology, it is known that Laishmaniasis causes Leishmania tropica (L. tropica) and Leishmania major (L. major) causes Cutaneous Laishmaniasis, Leishmania infantum (L. infantum) and Leishmania donovani (L. donovani) causes Visceral Laishmaniasis. In the recent years, molecular methods based on polymerase chain reaction (PCR) have been developed in the determination of genotypes and pathogenicity of Leishmania species. As a result of these studies L. tropica was isolated in VL cases without KL story, L. infantum species were isolated in KL cases without VL story. This situation reveals the probability that classical information reported in literature may be wrong, L. infantum and L. tropica may cause Laishmaniasis in both forms. In this study, L. tropica (49%), L. infantum (36%), L. major (15%) from Cutaneous Laishmaniasis; L. infantum (56%), L. donovani (31%) and L. Tropica (13%) species in Visceral Laishmaniasis were isolated. It was seen that L. İnfantum and L. major from Cutaneous Leishmaniasis were haplotype, L. tropica from Cutaneous Leishmaniasis was heterotype. It was found that L. infantum, L. donovani and L. tropica from Visseral Leishmaniasis were haplotype. The L. infantum’s nucleotide series has similarity with nucleotide series of L. İnfantum isolated from cases with Visceral Laishmaniasis. This situation makes us think that L. infantum species isolated from Cutaneous Laishmaniasis cases may cause Visceral Laishmaniasis. In the study L. tropica species were determined in Visceral Laishmaniasis cases which have no Cutaneous Laishmaniasis story. As a result of the study, it was determined that L. infantum which is known as the factor of Visceral Laishmaniasis can be observed in Cutaneous Laishmaniasis and that L. tropica which is known as the factor of Cutaneous Laishmaniasis can be observed in Visceral Laishmaniasis.Bu çalışma Ç.Ü. Bilimsel Araştırma Projeleri Birimi tarafından desteklenmiştir. Proje No: TF2010D11

Molecular epidemiology of Blastocystis

Blastocystis pathogenicity and classification was newly illuminated with molecular genetic studies and recently the parasite was found in the focus of many researchers. Several molecular methods such as; polymerase chain reaction (PCR), PCR-restriction fragment length polymorphism, random amplified polymorphic DNA, real-time polymerase chain reaction and DNA sequencing analyses can be used in genotyping of Blastocystis. Blastocystis parasites may cause diarrhea, abdominal pain, bloating, gas, irritability, anorexia, cramps, vomiting, dehydration, insomnia, nausea, loss of appetite, weight loss, fatigue symptoms and also could be asymptomatic cases. In this review, it was aimed to summarize the associations between Blastocystis subtypes and pathogenicity

Isolation of Cystic Echinococcosis Causative Agents of Echinococcus equinus and Echinococcus ortleppi Species from Humans in the Central Anatolia Region of Türkiye

Echinococcus granulosus sensu strico’nun neden olduğu kistik ekinokokkoz (KE) hastalığı, tarımın ön planda olması, sosyo-ekonomik durumun düşük olması ve hijyenik şartlarda olmayan hayvan kesimleri nedeniyle Türkiye’de çok sık görülen hastalıklardan biri olmasına rağmen ülkede ihmal edilmektedir. Has- talığın morbiditesi, mortalitesi ve tedavisinde yaşanan zorluklar göz önüne alındığında bu hastalıkla ilgili daha fazla çalışmalar yapılmasına ve önlemler alınmasına ihtiyaç duyulmaktadır. Bu çalışmada İç Anadolu bölgesindeki KE hastalarından izole edilen Echinococcus izolatlarını genotiplendirmek amaçlanmıştır. KE tanısı almış 60 hastadan alınan doku örneklerinden DNA izolasyonu QIAamp DNA FFPE doku (Qiagen, Hilden, Almanya) kiti kullanılarak yapılmıştır. Genotiplendirme için Echinococcus parazitinin cytochrome c oxidase subunit gen bölgesi hedef alınmış, JB3 ve JB4.5 primerleri kullanılmıştır. Polimeraz zincir reaksiyo- nu [polymerase chain reaction (PCR)] ürünleri QIAquick PCR pürifikasyon kitinin (Qiagen, Hilden, Alman- ya) kullanma talimatlarına göre saflaştırılmıştır. PCR ürünleri ABI Prism BigDye Terminator V3.1 Cycle se- quencing kiti (ThermoFisher Scientific, Litvanya) kullanılarak hazırlanmış ve örneklerdeki nükleotit dizileri ABI 3100 (Applied Biosystems, Birleşik Devletler) dizi analizi cihazı ile değerlendirilmiştir. Çalışmada elde edilen nükleotit dizileri, MetaPIGA2, MRBAYES v.3.1.2, “phyogenetic analysis using parsimony (PAUP v.4.0b10)”, Unigen programları, maksimum likelihood (ML), Bayesian (BI) ve parsimoni (P) yöntemleri kullanılarak filogenetik analiz yapılmıştır. Echinococcus izolatlarının %88.4 (53/60)’ünün E.granulosus s.s. olduğu bunların ise %41.7 (25/60)’sinin genotip 1, %30 (18/60)’unun genotip 3, %16.7 (10/60)’si- nin ise genotip 2 genotipinde olduğu saptanmıştır. Çalışmadaki diğer Echinococcus izolatlarının %6.6 (4/60)’sının E.equinus ve %5.5 (3/60)’inin E.ortleppi olduğu tespit edilmiştir. E.equinus ve E.ortleppi’nin atipik yerleşimli kistlerden ve kırsal bölgelerde yaşayan insanlardan izole edildiği görülmüştür. Kentsel böl- gelerde yaşayan KE hastalarında E.equinus ve E.ortleppi türlerine rastlanmamıştır. İç Anadolu bölgesinde köpek ve büyükbaş hayvancılık yetiştiriciliğine bağlı olarak KE olguları yaygın olup hastalık etkeni Echino- coccus türleri çeşitlilik göstermektedir. Echinococcus türlerinin genotiplendirilmesi KE tedavisi ve kontrol stratejilerinin gelişmelerinde etkilidir. Bu çalışmanın sonuçları son yıllarda dünyada ve Türkiye’de “tek sağlık” anlayışının temelini oluşturan KE mücadelesinde etkin rol oynabilir.Cystic Echinococcosis (CE) caused by Echinococcus granulosus sensu strico is neglected in Türkiye de- spite being one of the common diseases due to agriculture being at the forefront, low socioeconomic status and unhygienic animal slaughter. Considering the morbidity, mortality, and difficulties in treatment, more studies and precautions are needed regarding this disease. In this study, it was aimed to genotype Echinococcus isolated from CE patients in the Central Anatolia region. DNA isolation from tissue samples taken from 60 CE patients was performed using the QIAamp DNA FFPE tissue kit. Cytochrome c oxidase subunit gene region of Echinococcus was targeted and JB3/JB4.5 primers were used for genotyping. Poly- merase chain reaction (PCR) products were purified according to the instructions for use of the QIAquick PCR purification kit. PCR products were prepared using the ABI Prism BigDye Terminator V3.1 Cycle sequencing kit and the nucleotide sequences in the samples were evaluated with the ABI 3100 sequenc- ing device. The nucleotide sequences obtained in the study were analyzed using MetaPIGA2, MRBAYES v.3.1.2, phyogenetic analysis using parsimony, Unigen programs, maximum likelihood, Bayesian and par- simony methods. It has been found that 88.4% (53/60) of Echinococcus isolates were E.granulosus s.s. in this study. It has been genotyped as 41.7% (25/60) G1, 30.0% (18/60) G3 and 16.7% (10/60) G2 geno- type. It has been determined that 6.6% (4/60) of the other Echinococcus isolates were E.equinus and 5.5% (3/60) were E.ortleppi. It was observed that E.equinus and E.ortleppi were isolated from atypically located cysts and from those living in rural areas. The E.equinus and E.ortleppi species were not found in CE patients living in urban areas. CE cases are common in the Central Anatolia region due to dog and cattle breeding, and the disease agent Echinococcus species vary. Genotyping of Echinococcus species is effective in the de- velopment of CE treatment and control strategies. Study results can play an active role in the fight against CE, which has formed the basis of the “one health” approach in the world and in Türkiye in recent years

MicroRNA profile in immune response of alveolar and cystic echinococcosis patients

*Eroğlu, Fadime ( Aksaray, Yazar )It is known that miRNAs are effective in immune response in the diagnosis and treatment of many infectious diseases. However, the miRNAs profile is unknown in Alveolar and Cystic Echinococcosis which can be fatal if left untreated. The miRNAs profile that activates the T and B cells forming the immune system in Alveolar and Cystic Echinococcosis patients was investigated in this study. A total of 50 liver tissue samples were obtained from Alveolar and Cystic Echinococcosis patients in Kilis State Hospital Pathology Laboratory in southeast of Turkey. The circulating cell-free miRNAs were evaluated by a quantitative real-time polymerase chain reaction, statistically calculated within ΔΔCt values and fold changes were evaluated by Welch T test, in which P <.05 was considered to be significant. Twenty-five microRNAs, including let-7a-5p, let-7c, let-7e-5p, miR-15b-5p, miR16, miR-17-5p, miR-23a-5p, miR-24-3p, miR-25-3p, miR-26a-3p, miR-26b-3p, miR-29b-3p, miR-29c-3p, miR-30a-5p, miR-30b-5p, miR-30c-5p, miR-30d-5p, miR-30e-5p, miR-98-5p, miR-101-3p, miR-106b-5p, miR-125b-5p, miR-142-5p, miR-222-3p and miR-223-3p, were found as down-regulated in Alveolar and Cystic Echinococcosis patients than control groups. Twelve miRNAs, including miR-15a-5p, miR-21-5p, miR-27a-3p, miR-29a-3p, miR-146a-5p, miR-181a-5p, miR-181b-5p, miR-181d, miR-181c-5p, miR-195-5p, miR-214-3p and miR-365-3p, were found as up-regulated in Alveolar and Cystic Echinococcosis patients than healthy person. It has been shown that T- and B-cell activities are related in the progressive of both Alveolar and Cystic Echinococcosis in this study. The miRNA panel activated by T and B cells may be important for exploring the mechanisms underlying early development in Alveolar and Cystic Echinococcosis providing novel information that may be used to discover new therapeutics for these diseases

The molecular epidemiology of cystic and alveolar echinococcosis disease in southeast Turkey

Abstract Background: The migration of humans and climatic and environmental chang-es cause the emergence of infectious diseases. This study aimed to investigate the changes in the molecular epidemiology of the Echinococcosis disease in the southeast region of Turkey after migrations. Methods: Overall, 159 tissues samples were taken from suspected cases of Echinococcosis at the Kilis State Hospital in the southeast region of Turkey. All of the tissues samples were analyzed using histopathology methods, PCR, Re-al-time PCR methods, DNA sequencing, and phylogenetic analyses in labora-tories. Results: The positivity values of the histopathology, the polymerase chain reaction, and the Real-time PCR methods were found to be 14.5% (23/159), 15.7% (25/159), and 16.9% (27/159), respectively. 32.0 % (8/25) E. multilocu-laris of Echinococcus isolates and 68% (17/25) E. granulosus of Echinococcus iso-lates were identified using PCR methods. 58.8% (10/17) of the E. granulosus isolates were found to be Genotype 1% and 41.2% (7/17) E. granulosus isolates were found to be Genotype 3. Conclusion: Molecular methods play an important role in the epidemiology, treatment, and diagnosis of diseases. Increasing immigration in a geographical area may create social, economic, and health problems in that area. For this reason, epidemiological studies of infectious diseases should be updated in areas with immigration

Identification of leishmania tropica from pediatric visceral leishmaniasis in southern mediterranean region of Turkey

Background and Objective: Protozoa of the genus Leishmania are obligate intracellular parasites, and Leishmania species cause a spectrum of species-specific clinical symptoms known as cutaneous, mucocutaneous, and visceral leishmaniasis. For example, Leishmania major and Leishmania tropica cause cutaneous leishmaniasis, while Leishmania infantum and Leishmania donovani cause visceral leishmaniasis (VL). However, molecular studies in recent years have shown that Leishmania species cause different clinical symptoms. Objectives: Our aim was to evaluate the relationship between the clinical and molecular characterization of leishmania isolates in children with VL defined in Turkey, an intercontinental transitional region. Methods: The clinical diagnosis of VL was confirmed by detecting amastigotes in the bone marrow aspirate and/or the rK39 test and/or molecular methods (genus-specific PCR, Real-Time PCR, ITS1 PCR-RFLP, DNA sequencing). Results: Most of the VL patients were referred from the districts of Adana (53.3%) and others from neighboring provinces; Hatay (16.6%), Osmaniye (3%), Gaziantep (3%), Adiyaman (3%), and 20% case were Syrian immigrants A clinical diagnosis of VL was confirmed in 30 patients with different diagnostic methods. 93% was found positive with microscopic examination, 79.1% with rK39 dipstick test, and 60% with genus-specific PCR assay in clinical samples. The Leishmania isolates were identified as L. infantum (40%), L. donovani (26.7%), and L. tropica (23.3%) using Real-Time PCR, ITS1 PCR-RFLP, and DNA sequencing. There was no cutaneous finding in any case in clinical examination. The most common clinical findings were fever (93.3%) and splenomegaly (90%), followed by hepatomegaly (76.6%). The most common laboratory finding was thrombocytopenia (86.6%), followed by anemia (70%). In addition, hemophagocytic lymphohistiocytosis was detected in bone marrow aspiration in two of our patients. Since pentavalent antimony salts treatment initially failed in four patients, it was necessary to switch to Liposomal-Amphotericin B with treatment success. Conclusions: The presence of L. tropica in VL patients, despite the absence of cutaneous findings in any of the cases, shows that this strain can cause VL, contrary to conventional knowledge. In the Adana province, where this study was carried out, L. infantum from CL cases in previous studies should be taken into account, and visceral spread in CL cases and accompanying cutaneous lesions in VL cases should be investigated in detail