Sperm tyrosine kinase activity analyzes with immunocytochemistry

İstanbul Bilim Üniversitesi, Sağlık Bilimleri Enstitüsü, Histoloji ve Embriyoloji Anabilim Dalı Yüksek Lisans Programı.Akrozom reaksiyonu sonucu fertilizasyonun gerçekleşmesi spermlerin dişi genital sisteminde ilerlerken kazandıkları kapasitasyon yeteneğine bağlıdır. Kapasitasyon yeteneği kazanan ve hiperaktivasyon gösteren spermler akrozom reaksiyonu ve fertilizasyonu gerçekleştirebilirler. Fosfo tirozin kinaz aktivasyonunun kapasitasyon, akrozom reaksiyonu ve dolayısıyla fertilizasyon sürecinde etkili olduğu ve bu süreçte reaktif oksijen türevlerinin tirozin kinaz aktivasyonunu etkilediği bildirilmiştir. Çalışma, semende reaktif oksijen türevi kaynağı lökositlerin artmış olduğu lökospermi vakaları ile normospermili vakalardan seçilen iki grupta gerçekleştirildi. Ayrıca her iki grup, sperm hazırlama yönteminde kullanılan santrifüj işlemi sonucu ortaya çıkan reaktif oksijen türevlerinin olası etkilerinin değerlendirilmesi amacıyla iki alt gruba ayrıldı. İkiye ayrılan semen örnekleri santrifüj işleminin uygulanmadığı swim-up ve santrifüj uygulanan konsantrasyon yöntemleriyle hazırlandı ve immünositokimya için sperm yaymaları hazırlandı. Hazırlanan örnekler streptavidin-biotin peroksidaz yöntemiyle fosfo tirozin monoklonal antikoruyla işaretlendi. Yapılan değerlendirmelerde sperm tirozin kinaz aktivasyonunun spermlerin kuyruk bölgesinde lokalize olduğu, vakaların ekspresyon seviyelerinin bireysel farklılıklar gösterdiği izlendi. Normospermili vakalarda konsantrasyon grubunda swim-up grubuna göre aktivasyonun arttığı ancak bu artışın anlamlı olmadığı izlendi. Lökospermili vakalarda da konsantrasyon yöntemi uygulanan grupta bir artış gözlendi. Normospermili ve lökospermili gruplar arasında ise anlamlı bir fark tespit edilmedi. Normo ve lökospermili vakalarda reaktif oksijen türevlerinin sperm tirozin kinaz aktivitesi üzerine etkili olduğu söylenebilirse de bireysel farklılıkları ve sonuçlarını açıklayacak ileri çalışmaların yürütülmesi gerekmektedir.Acrosome reaction and fertilization depends on the capacitation ability gained by the sperms while in the female reproductive tract. Sperms that gain capacitation ability and show hyperactivity can realize acrosome reaction and fertilization. It is reported that phospho tyrosine kinase activation is effective in achieving capacitation, acrosome reaction and hence in fertilization process and reactive oxygen species have an effect in achieving tyrosine kinase activation during this process. This study was conducted in two groups by selecting cases of normospermia and cases of leukocytospermia with increased levels of leukocytes due to reactive oxygen species in semen. Additionally both groups have been separated into two subgroups each, in order to evaluate the effects of reactive oxygen species that appear in the centrifuge operation used during sperm preparation process. Semen samples that were separated into two were prepared by using swim up method where centrifuge process was not used and concentration method where centrifuge process was used and sperm smears were prepared for immunocytochemistry. The prepared samples were marked with anti-phospho tyrosine monoclonal antibody by using streptavidin-biotin peroxidase method. In the evaluation process, it was observed that sperm tyrosine kinase activation was localized in the tail section of the sperms, and expression levels of each case showed individual variances. Among the normospermia cases, it was observed that the activation increased in the concentration group in relation to the swim-up group, but this increase was not significant. In the leukocytospermia cases also the concentration group had increased activation. No significant differences were observed between normospermia and leukocytospermia groups. Even though it can be said that both among the normospermia cases and leukocytospermia cases, reactive oxygen derivatives are effective in sperm tyrosine kinase activity, further studies should be held in order to explain individual differences and results

Prenatal diagnosis of Comel-Netherton syndrome with PGD, case report and review article

İstanbul Bilim Üniversitesi, Tıp Fakültesi.Introduction Comel-Netherton syndrome (C-NS) is an autosomal recessive disorder of the skin, hair and immune system first reported by Comel in 1949 and Netherton in 1958 (1, 2). This syndrome presents at or soon after birth with generalized erythroderma, scaling, and/or continuous peeling of the skin resembling nonbulloous congenital ichthyosiform erythroderma or peeling skin syndrome. In the neonatal period, 20% of the babies suffer from hypernatremic dehydration, electrolyte imbalances, perturbed thermoregulation, failure to thrive and recurrent infections which may result in neonatal demise [3–5]. The skin lesions are often pruritic, resemble atopic eczema, and show an unstable, undulating course. They are usually accompanied by hair shaft abnormalities that develop during early childhood and may result in diffuse alopecia. The hallmark of C-NS is trichorrhexis invaginata (bamboo hair), but other abnormalities, including pili torti (twisted hair) and trichorrhexis nodosa (hair of varying diameter) have been observed. Markedly elevated IgE levels, allergic reactions to food and common antigens, malnutrition, and increased susceptibility to skin, respiratory tract or systemic infections are also characteristic [6, 7]. There are nearly 150–160 cases of C-NS reported in the literature, its incidence might be 1/200.000 [6] due to challenging diagnostic problems during infancy and early childhood. This syndrome has overlapping features with atopic dermatitis and other recessive ichthyosis. Most patients with C-NS are sporadic cases however, there are reports of affected siblings and of consanguinity in about 10% of the families with C-NS which is common in autosomal recessive inheritance [8–10]. In this study, we present two consanguineous Turkish families (two sisters married with two brothers) of prenatal diagnosis of Netherton syndrome and successful intracytoplasmic sperm injection (ICSI) pregnancies using PGD and a review of literature. To our knowledge, this is the first report of PGD and ICSI in Comel-Netherton syndrom