Targeted high-throughput sequencing for genetic diagnostics of hemophagocytic lymphohistiocytosis

Background: Hemophagocytic lymphohistiocytosis (HLH) is a rapid-onset, potentially fatal hyperinflammatory syndrome. A prompt molecular diagnosis is crucial for appropriate clinical management. Here, we validated and prospectively evaluated a targeted high-throughput sequencing approach for HLH diagnostics. Methods: A high-throughput sequencing strategy of 12 genes linked to HLH was validated in 13 patients with previously identified HLH-associated mutations and prospectively evaluated in 58 HLH patients. Moreover, 2504 healthy individuals from the 1000 Genomes project were analyzed in silico for variants in the same genes. Results: Analyses revealed a mutation detection sensitivity of 97.3 %, an average coverage per gene of 98.0 %, and adequate coverage over 98.6 % of sites previously reported as mutated in these genes. In the prospective cohort, we achieved a diagnosis in 22 out of 58 patients (38 %). Genetically undiagnosed HLH patients had a later age at onset and manifested higher frequencies of known secondary HLH triggers. Rare, putatively pathogenic monoallelic variants were identified in nine patients. However, such monoallelic variants were not enriched compared with healthy individuals. Conclusions: We have established a comprehensive high-throughput platform for genetic screening of patients with HLH. Almost all cases with reduced natural killer cell function received a diagnosis, but the majority of the prospective cases remain genetically unexplained, highlighting genetic heterogeneity and environmental impact within HLH. Moreover, in silico analyses of the genetic variation affecting HLH-related genes in the general population suggest caution with respect to interpreting causality between monoallelic mutations and HLH. A complete understanding of the genetic susceptibility to HLH thus requires further in-depth investigations, including genome sequencing and detailed immunological characterization.Peer reviewe

ÇOCUKLUK ÇAĞI LÖSEMİLERİNDE VE NÖTROPENİK ATEŞ ATAĞI SIRASINDA NEOPTERİN DÜZEYLERİ

Çalışmamızda, makrofajlardan salınarak hücresel bağışıklıkta rol oynayan neopterinin; çocukluk çağı lösemilerinde tanı, indüksiyon tedavisi ve nötropenik ateş sırasındaki düzeyleri araştırıldı. 20 ALL/5AML (0-18 yaş) olgusu ile &#8216;Yeni Tanı Grubu&#8217; oluşturuldu. İndüksiyon tedavisinin 0, 15, 33. günlerinde; kan alınarak ELISA yöntemiyle çalışıldı. Neopterin düzeyleri beyaz küre (BK), mutlak nötrofil (ANS), mutlak fagosit sayısı (AFS), CRP düzeyleri ve kemik iliği aspirasyon bulgularıyla karşılaştırıldı. Hem ALL, hem AML grubunda neopterin düzeylerinin indüksiyon tedavisi boyunca diğer parametrelerle ve kemik iliği blast yüzdesi ile uyumlu olarak düştüğü saptandı (p<0,05). Çocukluk çağı lösemi olgularında nötropenik ateş atağı sırasında neopterin düzeyleri, 19 hastadaki(1-18 yaş) 30 nötropenik ateş atağında değerlendirildi. Ateşin 0, 3, 5. günlerinde neopterin düzeyleri; BK, ANS, AFS, CRP düzeyleri ile karşılaştırıldı. ANS, AFS düzeylerinde 3-5. günler arasında artış görülürken (p0,05). Ateş atakları klinik (n:15); mikrobiyolojik tanılı enfeksiyon (n:7) ve odağı bulunamamış ateş (n:8) olarak üç gruba ayrıldı. Gruplar arasında neopterin düzeyi farklı saptanmadı.. Söz edilen iki grup ile enfeksiyon kontrol grubu (n:19) (1-18 yaş) ve sağlam çocuk kontrol grubunun (n:21) (6-18 yaş) neopterin düzeyleri karşılaştırıldı. Yeni tanı grubunda 7,03 ng/ml (0,1-17,04), nötropenik ateş grubunda 0,56 ng/ml (0,1- 21,85), enfeksiyon kontrol grubunda 7,02 ng/ml (0,1-27,22) ve sağlam çocuk kontrol grubunda 0,1 ng/ml (0,1-4,29) olarak saptandı. Yeni tanı ve enfeksiyon kontrol grubunun neopterin değerlerinin en yüksek ortanca değer oldukları 137 görüldü. Nötropenik ateş grubunun neopterin düzeyi sağlam çocuk kontrol grubuna göre yüksek bulundu (p<0,05). Çalışmamız, çocukluk çağı lösemilerinde tanı anında neopterin düzeyinin yüksek olup, hastalık aktivitesini göstermede kullanılabileceğini düşündürdü

The usage of pediatric bleeding questionnaire in the diagnosis of von willebrand disease and thrombocyte function defects among Turkish children

Çocukluk çağında hafif derecedeki kanama diyatezlerinin tanısı sağlam olguların da benzer semptomlar bildirmesiyle zor olmaktadır. Bu olgularda tanıya ulaşmada ayrıntılı ve doğru bir kanama öyküsünün elde edilmesiyle standardize pediatrik kanama skoru geliştirilmiştir. Çalışmamızda Türk çocuklarında pediatrik kanama skoru daha önceden tanı almış Von Willebrand Hastalığı (VWH) (n=46), Trombosit Fonksiyon Bozukluğu (TFB) (n=65) ve yakınması olmayan sağlam `Kontrol 1'(n=38) ve kanama yakınması olan ancak hemostatik olarak normal bulunan `Kontrol 2'(n=32) gruplarına uygulandı. Pediatrik kanama skorunun (PKS) cutt off değeri ? 3 olarak saptandı (AUC=0.785 %95 CI:0.718-0.852). Von Willebrand Hastalığı grubu ve Kontrol 1 grubunu ayırt etmede sensitivite, spesifite, Pozitif Prediktif Değer (PPD) ve Negatif Prediktif Değer (NPD) sırasıyla %93,5; %97,4; %97,7; %92,5 saptandı. Von Willebrand Hastalığı grubu ve Kontrol 2 grubunu ayırt etmede ise sensitivite, spesifite, PPD ve NPD sırasıyla %93,5; %87,5; %91,5;%90,3 saptandı. Sensitivite, spesifite, PPD ve NPD sırasıyla TFB ve Kontrol 1 grubunu ayırt etmede %89,2; %97,4; %98,3; %84,1; TFB ve Kontrol 2 grubunu ayırt etmede %89,2; %87,5; %93,5; %80 olarak saptandı. Sonuç olarak, PKS'nin kanama diyatezi ile sağlam olguları ayırt etmede iyi bir araç olduğu ancak kesin tanı için ek hemostatik laboratuar tetkiklerinin gerekli olduğu düşünüldü.The diagnosis of mild bleeding disorders is not easy in the pediatric population as most of the `healthy?subjects also report bleeding symptoms. In order to get a precise bleeding history Pediatric Bleeding Questionnaire (PBQ) has been developed. In our study, Turkish children previously diagnosed with Von Willebrand Disease (VWD), Thrombocyte Function Defect (TFD) and healthy children without any symptoms (Control group 1) and healthy children with symptoms but found hemostatically normal (Control group 2) were analysed with PBQ. The cutt off level was found to be ? 3. The sensitivity, specifity, PPV and NPV of PBQ to define VWD vs. Control group 1 was %93.5; %97.4; %97.7. %92.5; VWD vs. Control group 2 was %93.5; %87.5; %91.5;%90.3; TFD vs. Control group 1 was %89.2; %97.4; %98.3; %84.1 and TFD vs. Control group 2 was %89.2; %87.5; %93.5; %80 respectively. In conclusion, PBQ was found to be useful in the diagnosis of VWD and TFD in Turkish children, however to reach an exact diagnosis hemostatic laboratory tests are stil warranted

Type I allergic hypersensitivity reactions due to ethylene oxide sterilised leucocyte filters in patients with thalassaemia: report of four cases.

Ethylene oxide (EO) is a highly reactive gas used in sterilisation of heat sensitive medical devices, such as infusion sets, cannulae, intubation materials, ventriculoperitoneal shunts, dialysis catheters and stents. Allergic reactions due to EO have been reported in haemodialysis patients, patients undergoing extracorporeal photopheresis and donors of plasmapheresis. Clinical manifestations vary considerably and generally do not allow differentiation between IgE-mediated anaphylaxis and anaphylactoid reactions. We report four patients with thalassaemia who experienced anaphylaxis during transfusion due to ethylene oxide sterilised leucocyte filters. The aim of this report is to highlight the fact that frequently transfused patients can have allergic reactions due to EO particles left in leucocyte filters

Adaptation and validation of Turkish version of musculoskeletal pain intensity and interference questionnaire for musicians

Adaptation and validation ofTurkish version of musculoskeletal painintensity and interference questionnaire formusiciansBS Akel, Ö BelenHacettepe University, Ankara, Turkey. Objective: There are certain pain questionnaires orscales that are translated and adapted to Turkish;however, we observed an absence of an assessmenttool to examine pain in Turkish-speaking musicians.The Musculoskeletal Pain Intensity and InterferenceQuestionnaire for Musicians (MPIIQM) evaluated notonly the severity of pain, but also the impact of pain onquality of life and the experience of playing the instrument. The aim of this study was to translate and adaptthe MPIIQM in Turkish; and to test its validity and reliability for Turkish-speaking musicians whom havepain and pain-related music-playing issues.Methods: The MPIIQM was translated into Turkish bytwo independent native Turkish speakers. Translationswere compared for inconsistencies and aggregatedinto a single Turkish version. This version then alsowas back-translated into English by two independentnative English speakers. After the back-translationswere compared for inconsistencies and aggregatedinto a single form, the final English version and the original questionnaire were also compared for inconsistencies. Finally, the original English questionnaireand the Turkish questionnaire was reviewed by abilingual team, to check for the errors of interpretation and nuances that might have been missed. TheTurkish questionnaire was finalized after consensus.The study was conducted on 60 professional musicians whom had pain-related playing issues. TheMcGill Pain Questionnaire and Disabilities of Arm,Shoulder and Hand questionnaire (DASH) were alsoadministered, within an interval of 7 days (retest).Instrument test-retest reliability was assessed withthe interclass correlation coefficient (ICC) and withthe Pearson’s correlation coefficient.Results: Translation and back-translation revealedno major difficulties. Reliability of the Turkish versionof the questionnaire was very good, with high consistency and reproducibility. The MPIIQM Turkish versionshowed a high correlation with the DASH and McGillquestionnaires.Conclusions: In conclusion, the results displayed thatthe Turkish version of the MPIIQM is a reliable andvalid region-specific version and proper for use onmusicians. It seemed to be a reliable, consistent andvalid instrument in evaluating the pain intensity andimpact of pain on musicians.</p

Infection associated hemophagocytic syndrome in children: a retrospective analysis of 15 cases

Amaç:Hemofagositik sendrom (HFS) aşırı immün aktivasyona bağlı hızlı ilerleyen ve hayatı tehdit eden birsendromdur. HFS ile ilişkili klinik tablo ve laboratuvar anormalliklerinin erken tanınması ve tedaviye hemenbaşlanması hayat kurtarıcı olabilmektedir. Bu çalışmada enfeksiyon ilişkili hemofagositik sendrom (EİHFS)tanısıyla izlediğimiz çocuk hastalarda; altta yatan enfeksiyöz etkenlerin, klinik, laboratuvar bulguların ve tedavisonuçlarının incelenmesi amaçlanmıştır.Gereç ve yöntem: Aralık 2012 ile Ocak 2016 arasında EİHFS tanısı konan çocuk hastaların kayıtları geriyedönük olarak incelendi.Bulgular: Çalışmaya yaşları ortanca 6 yaş (2-16 yaş) olan toplam 15 hasta (8 erkek, 7 kız) dahil edildi. HFS’yitetikleyen en sık enfeksiyöz etken olarak Brusella (8 hasta, %53) saptandı. Diğer altta yatan enfeksiyöz etkenler;3 hastada Leishmania, 1 hastada Salmonella Typhi, 1 hastada Mycobacterium tuberculosis, 1 hastada parvovirüsB19 ve 1 hastada influenza A (H3N2) idi. Ateş, bi/pansitopeni, hiperferritinemi ve serum transaminaz düzeylerindeyükseklik tüm hastalarda (%100), splenomegali 12 hastada (%80) mevcuttu. Tüm hastaların kemik iliği aspirasyonincelemesinde hemofagositoz görüldü. Tüm hastalara altta yatan enfeksiyonuna yönelik antimikrobiyal tedavi;uzamış ateş ve klinik durumunda giderek kötüleşme olan 8 hastaya intravenöz immünglobülin (İVİG), 2 hastayaİVİG+steroid tedavileri verildi. Olguların %93’ünde (14 hasta) iyileşme kaydedildi. İnfluenza A (H3N2) pnömonisiolan sadece 1 hastada (%6) ölüm görüldü.Sonuç: Hekimler, enfeksiyon nedeniyle izlenen hastalarda uzamış ateş, splenomegali ve sitopeninin varlığındaHFS gelişimi konusunda farkındalığa sahip olmalıdır

Peripheral Neuropathy: Not a Feature of Childhood Thalassemia

Background: Chronic anemia in thalassemia patients may cause multiple complications such as bone deformities, growth retardation, and peripheral neuropathy. Aim: To examine the presence of possible electrophysiological changes in children diagnosed with thalassemia and to investigate the clinical factors affecting the electrophysiological findings in those children. Subjects and Methods: A hospital based prospective study. This prospective study included 154 children, who were diagnosed as having thalassemia and 100 control cases. Demographic features and laboratory data were recorded. All cases were examined electrophysiologically using standard procedures. Results: Totally 154 patients and 100 control cases were included in the study. Neurological examination did not indicate any abnormalities in any of the participants. Any evidence of large-fiber neuropathy was not present in any of the participants. Conclusions: In this study, we did not find any cases with neuropathy. With these results we can conclude that, thalassemia at early stages is not a risk factor for polyneuropathy in thalassemia patients under follow-up

Diagnostic Value of Neopterin during Neutropenic Fever and Determination of Disease Activity in Childhood Leukemias

Neopterin, a pteridine group compound that is secreted from macrophages is shown to be increased in adult leukemia; however there are few studies in childhood leukemia. This study aimed to investigate neopterin levels during childhood leukemia treatment and neutropenic fever episodes for the possibility of using as a marker for disease activity and differentiation of infections