Background Severe congenital neutropenia is a rare disease, and autosomal dominantly inherited ELANE mutation is the most frequently observed genetic defect in the registries from North America and Western Europe. However, in eastern countries where consanguineous marriages are common, autosomal recessive forms might be more frequent. Method Two hundred and sixteen patients with severe congenital neutropenia from 28 different pediatric centers in Turkey were registered. Results The most frequently observed mutation was HAX1 mutation (n = 78, 36.1%). A heterozygous ELANE mutation was detected in 29 patients (13.4%) in our cohort. Biallelic mutations of G6PC3 (n = 9, 4.3%), CSF3R (n = 6, 2.9%), and JAGN1 (n = 2, 1%) were also observed. Granulocyte colony‐stimulating factor treatment was given to 174 patients (80.6%). Two patients died with infectious complications, and five patients developed myelodysplastic syndrome/acute myeloblastic leukemia. The mean (± mean standard error) follow‐up period was 129.7 ± 76.3 months, and overall survival was 96.8% (CI, 94.4–99.1%) at the age of 15 years. In Turkey, severe congenital neutropenia mostly resulted from the p W44X mutation in the HAX1 gene. Conclusion In Turkey, mutation analysis should be started with HAX1, and if this is negative, ELANE and G6PC3 should be checked. Because of the very high percentage of consanguineous marriage, rare mutations should be tested in patients with a negative mutation screen.

Objective: Atherosclerosis is a chronic inflammatory condition and is one of the main causes of death worldwide. Macrophages play important roles in the formation of atherosclerotic plaques. Apoptosis is progressively observed while plaques develop, although the precise mechanisms and outcomes of apoptosis in atherosclerosis development and progression are still contradictory. This study was conducted to explore the effects of simvastatin and retinoic acid receptor-related orphan receptor alpha (RORa) ligands on apoptosis in human acute monocytic leukemia (THP-1) macrophage cells. Methods: Briefly, the occupancy of RORa in the promoter regions of apoptotic pathway genes was demonstrated in THP-1 cell lines using chromatin immunoprecipitation (ChIP) analysis. In order to modulate RORa activity, THP-1 macrophage cells were treated with specific ligands (CPG52608 and SR1001) and then viability as well as count of THP-1 macrophage cells were analyzed. Results: We observed that simvastatin and both RORa ligands had a tendency to decrease THP-1 macrophage cell viability in culture. When compared with non-treated controls, simvastatin significantly decreased cell viability (p=0.04) and cell count (p=0.03). However, this negative effect of simvastatin seemed to be partly prevented by RORa ligands. In addition, bioinformatics analysis of ChIP-on-chip data demonstrated that several genes that are involved in the apoptotic pathway were likely RORa target genes. These genes were involved in the regulation of apoptosis through various pathways. Conclusion: In summary, our study suggest that simvastatin-mediated macrophage apoptosis might be modulated by SR1001 administration. However, involvement of RORa in this modulation through potential apoptotic target genes remains elusive.

The scope of cell-free DNA (cfDNA) testing was expanded to the genome, which allowed screening for rare chromosome anomalies (RCAs). Since the efficiency of the test for RCAs remains below the common aneuploidies, there is a debate on the usage of expanded tests. This study focuses on the confirmatory and follow-up data of cases with positive cfDNA testing for RCAs and cases with screen-negative results in a series of 912 consecutive cases that underwent invasive testing following cfDNA testing. Chorion villus sampling (CVS), amniocentesis (AS), fetal blood sampling, and term placenta samples were investigated using classical cytogenetic and molecular cytogenetic techniques. Out of 593 screen-positive results, 504 (85%) were for common aneuploidies, 40 (6.7%) for rare autosomal trisomies (RATs), and 49 (8.3%) for structural chromosome anomalies (SAs). Of the screen-positives for RATs, 20 cases were evaluated only in fetal tissue, and confined placental mosaicism (CPM) could not be excluded. Among cases with definitive results (n = 20), the rates of true positives, placental mosaics, and false positives were 35%, 45%, and 10%, respectively. Among screen-positives for SAs, 32.7% were true positives. The confirmation rate was higher for duplications than deletions (58.3% vs. 29.4%). The rate of chromosomal abnormality was 10.9% in the group of 256 screen-negatives with pathological ultrasound findings. This study provides further data to assess the efficiency of expanded cfDNA testing for RATs and SAs. The test efficiency for cfDNA seems to be higher for duplications than for deletions, which is evidence of the role of expert ultrasound in identifying pregnancies at increased risk for chromosome anomalies, even in pregnancies with screen-negatives. Furthermore, we discussed the efficiency of CVS vs. AC in screen-positives for RATs.

ÖZGelişim evrelerinin veya dokuya özgü fizyolojik süreçlerin düzenlenmesinde ve yürütülmesinde farklılaşmış gen ekspresyonu büyük önem taşımaktadır. "Subtractive" hibridizasyon yöntemi dokuya özgü ekspresyonu olan genlerin tespit edilmesi için oldukça etkin bir yöntemdir. Daha önceki çalışmalarda laboratuvarımızda BALB/c ırkı farelerde kalbe özgü ekspresyonu olan genleri içeren "Subtractive" hibridizasyon cDNA kütüphanesi oluşturulmuştur. Önemli bir transkripsiyon faktörü olan Taube Nuss (Tbn), bu kütüphaneden elde edilen genlerden birisidir. Bu çalışmadaki amacımız, Tbn geni ile ilişkili olduğu düşünülen diğer genlerin ekspresyonlarının araştırılması ve Tbn yolağının belirlenmesidir. Çalışma kapsamında BALB/c ırkı farelerden elde edilen kalp ve iskelet kası dokularına özgü Tbn geninin ekspresyonu Northern Blot analizi ile araştırılmıştır. Ardından, Tbn genine özgü siRNA'lar kullanılarak H9c2 rat kardiyomiyoblast hücre soyunda Tbn geni sessizleştirilmiştir. Yolak analizleri sonucunda Tbn ile ilişkili olabileceği tespit edilen Pparg, Cebpa, Taf10 ve Myocd genlerinin sessizleştirilen hücrelerdeki ekspresyon değişimleri araştırılmıştır. Bu hücrelerde, Myocd gen ekspresyonunun azaldığı ve Pparg gen ekspresyonunun arttığı saptanmıştır. Bu sonuçlar, Tbn geninin kalp dokusunda düzenleyici bir rol oynadığını işaret etmektedir. ABSTRACTDifferential gene expression is important in the regulation and maintenance of developmental stages and tissue specific physiological processes. The subtractive hybridization method is an efficient method for the detection of genes that have tissue specific expressions. In our previous work, our team had constructed a BALB/c mice heart tissue specific subtractive hybridization cDNA library. Taube Nuss (Tbn), which is a crucial transcription factor, is one of the genes that were isolated from this library. In this study, our aim was to analyze the expression of possible Tbn related genes and identify the Tbn pathway. In the scope of this study, the Tbn gene expression in BALB/c mice heart and skeletal tissue was analyzed with the Northern Blot technique. Then, the Tbn gene was silenced in H9c2 rat cardiomyoblast cell line using siRNA specific to the TBN gene. The gene expression levels of Pparg, Cebpa, Taf10 and Myocd genes, which were possibly related to the Tbn pathway, were analyzed in silenced cells. We observed that the expression of the Myocd gene was decreased, whereas the expression of the Pparg gene was increased in these cells. These results suggest that the Tbn gene plays a regulatory role in cardiac tissue.

Yardımla üreme tedavileri sırasında, insan preimplantasyon embriyoları genellikle in vitro kültür ortamı, hasta yaşı, ovarian stimülasyon protokolleri ve gametlere bağlı nedenlerle gelişimin çeşitli aşamalarında duraklayabilmektedir. Gelişimsel duraklamanın (arrest) gözlendiği çiftlerde normal gelişim gösteren diğer embriyoların transfer edilmesinden sonra implantasyon başarısızlığı ve düşük gözlenebilmektedir. Diğer yandan, embriyonik arrestin altında yatan nedenler kesin olarak açıklanabilmiş değildir. Bu çalışmada amacımız, erken dönem embriyonik arrestin yardımla üreme teknikleri başarısı üzerine etkisinin araştırılmasıdır.Gereç ve Yöntem: Bu retrospektif çalışma kapsamında, 2010 ve 2018 yılları arasında yardımla üreme tedavisi gören ve ovulatuvar faktör, tubal faktör, endometriozis, açıklanamayan infertilite, ve erkek faktörü tanısı konulmuş olan 620 infertil çiftin intrasitoplazmik sperm enjeksiyonu-embriyo transferi (ICSI-ET) siklusu değerlendirilmiştir. Çalışma grupları, embriyonik arrest gözlenen hastalar ve tüm embriyoları duraklamadan normal hızda gelişim gösteren (kontrol) hastalardan oluşmaktadır. Çalışma datası, tek-yönlü değişken analizi (ANOVA) ve Ki-kare testi kullanılarak analiz edilmiştir.Bulgular: Çalışma toplumunda, kullanılan ilaç dozu, oosit sayısı, fertilizasyon oranı, iyi kaliteli (GI) ve düşük kaliteli (GIII) embriyo oranları anlamlı olarak arrest yönünde artmışken, gebelik oranlarının azaldığı (p<0,0001) ve canlı doğum oranlarında iki grup arasında bir farklılığın olmadığı (p=0,449) tespit edilmiştir.Sonuç: Embriyo kohortunda arrest embriyoların bulunmasının yardımla üreme teknikleri başarısı üzerine olumsuz etkilerinin olduğu çeşitli çalışmalarda belirtilmiştir. Çalışmamızda elde edilen bulgular bu doğrultudadır.