Molecular and Genetic Medicine

Background and objectives: The objective of this study is to determine the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in 201 Turkish patients who were referred to our clinic with venous thrombosis complications such as deep venous thrombosis, ischemic complications, thromboembolism and coronary artery disease. Methods: After isolation of genomic DNA from peripheral blood samples, polymerase chain reaction (PCR) and restriction fragment length polymorphism techniques were used for analysis. Results: Among patients with venous thrombosis complications, allelic frequencies were 0.33, 0.17 and 0.04 for MTHFR (C677T), factor V Leiden (G1691A) and prothrombin (G20210A) mutations respectively. Conclusion: Homozygosity for the MTHFR C677T mutation and/or presence of at least one copy of the A allele of the Factor V Leiden G1691A mutation was found to be associated with increased incidence of venous thrombosis complications in patients (p<0.01). The combined impact of these mutations on venous thrombosis should also be taken into consideration. In our study, prothrombin (G20210A) mutation was found not to be associated with venous thrombosis complications. We also found that the prevalence of factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in Turkish patients with venous thrombosis are comparable to results of other studies performed in Turkish and Caucasian populations. We did not observe any significant gender dependency for the factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations in venous thrombosis complications.

Casein kinase 2 (CK2) is an oncogenic protein kinase which contributes to tumor development, proliferation, and suppression of apoptosis in multiple cancer types. The mechanism by which CK2 expression and activity leads to tumorigenesis in glioblastoma (GBM), a stage IV primary brain tumor, is being studied. Recent studies demonstrate that CK2 plays an important role in GBM formation and growth through the inhibition of tumor suppressors and activation of oncogenes. In addition, intriguing new reports indicate that CK2 may regulate GBM formation in a novel manner; CK2 may play a critical role in cancer stem cell (CSC) maintenance. Since glial CSCs have the ability to self-renew and initiate tumor growth, new treatments which target these CSCs are needed to treat this fatal disease. Inhibition of CK2 is potentially a novel method to inhibit GBM growth and reoccurrence by targeting the glial CSCs. A new, orally available, selective CK2 inhibitor, CX-4945 has had promising results when tested in cancer cell lines, in vivo xenograft models, and human clinical trials. The development of CK2 targeted inhibitors, starting with CX-4945, may lead to a new class of more effective cancer therapies.

In this work we are presenting our novel adaptation of flow cytometry for the determination of change of diplococcic Gram-positive Streptococcus mitis population heterogeneity by their xylitol metabolism. The inherent population heterogeneity of the bacteria due to their growth in-chains of varying lengths was grouped into three different groups by flow cytometric analysis, designated as gates P1, P2 and P3. Gate P1 consists of the bacterial subpopulation with minimum cell wall thickness and therefore the least side scattering, while gate P3 consists of the subpopulation with the most side scattering (corresponding to the thick bacterial cell wall). According to our results gate P1 contains bacteria in long-chains, while P3 contains individual bacteria. When these diplococcic bacteria were grown in the presence of 2% xylitol more homogeneous population was seen as P1 gate was populated with approximately 80% of the total population. These results suggest that once xylitol is metabolized by these bacteria, they stabilize in long chains. These ‘xylitol stabilized’ long chains arose apparently by incomplete bacterial separation, contains individual bacteria with reduced cell wall thickness, resulted less side scattering. The optimum population homogeneity was achieved when these bacteria were grown in 2% xylitol and 300 ppm fluoride containing medium, as measured by the bacterial population percentage in gate P1. Since these bacteria in long chains are known to be in a latent phase, our findings demonstrate that if appropriately developed xylitol has potential to use as an alternative antimicrobial for life threatening diseases.

It has long been envisaged that gene disruption or gene correction in affected target cells can be efficiently conducted in vitro and in vivo and over the recent years several tools for achieving this goal were developed. Designer nucleases such as zinc finger nucleases (ZFNs) were extensively explored and more recently transcription activator-like effector nucleases (TALENs) were introduced for sequence-specific genome engineering in the mammalian genome. ZFNs and TALENs are fusion proteins containing a customized DNA-binding motif for sequence-specific DNA binding linked to a nuclease for introduction of double-stranded DNA breaks. Both systems were explored in mammalian cells using non-viral and viral delivery methods. Herein, we will provide a state-ofthe- art overview of available virus-based delivery systems for sufficient expression of functional TALENs. We will cover the molecular design of recombinant viruses containing TALEN expression cassettes and we will mention advantages and disadvantages of the respective systems. Although the relevance of these viral vector systems for novel developments in molecular medicine and genome engineering need to be further evaluated, we believe that with further improvements these viral vectors for TALEN delivery will play an emerging role in bioengineering and for establishing novel therapeutic concepts.

Objective: To investigate the ability of Cell-Free RNA BCT (BCT) blood collection device to stabilize fetal cellfree RNA in maternal plasma when compared to K3EDTA collection tubes. Design and methods: Blood samples were drawn from healthy pregnant donors into K3EDTA tubes and BCTs and kept at ambient temperature (22°C). Plasma was separated by centrifugation and cell-free RNA was extracted. Circulating fetalRNAs from plasma were quantified by digital droplet polymerase chain reaction (ddPCR). Results: Blood drawn into K3EDTA tubes showed a decrease in fetal mRNA concentration for human placental lactogen (hPL), β subunit of human chorionic gonadotropin (βhCG) and placenta-specific 4 (PLAC4) over three days of ex vivo incubation at 22°C. Blood drawn into BCTs, however, showed no significant change in mRNA concentrations over the same period of time. Conclusion: Cell-Free RNA BCT blood collection devicepreservescirculating fetal RNA in maternal blood for at least for three days at 22°C, which enhances the potential clinical utility ofcirculating fetal RNA in maternal blood for noninvasive prenatal diagnostic assay development.

Nuclear factor κB (NF-κB) is a family of transcription factors that play central roles in multiple biological functions, including immune and inflammatory responses and tumorigenesis. The regulation of the activity of NF-κB is very sophisticated. Post-translational modifications of NF-κB have long been known to play an essential role in the regulation of NF-κB activity. Since the p65 subunit (RelA) is the major subunit of the typical NF-κB heterodimer, more attention has been recently devoted to how p65 is regulated by the post-translational modifications. In this review, we provide an overview of the most recent developments in the regulation of p65 by post-translational modifications.

c-Jun activation domain-binding protein 1 (JAB1) was originally identified as a c-Jun co-activator and subsequently discovered to be a component of the COP9 signalosome (CSN) complex. JAB1, which is also known as CSN5, affects many partner proteins through protein-protein interaction, which leads to protein degradation and transcriptional activation. Thereby, JAB1/CSN5 functions as a multifunctional protein involved in the regulation of cell cycle, signal transduction, and DNA repair. In particular, JAB1/CSN5 plays an essential role in tumorigenesis by degrading tumor suppressor proteins and activating oncogenic transcription factors. JAB1/CSN5 overexpression has been observed in various types of cancer, and it has been multifunctionally involved in cancer progression. In this review, we provide an overview of the roles of JAB1/CSN5 in tumorigenesis and summarize recent findings that highlight the novel roles of JAB1/CSN5 in this process

Objectives: Most times in the medical practice, when we have a case of bone fracture, the patient is sent to the Orthopedic Department in order to treat locally the bone fracture with bone consolidation, without thinking at other diseases, of which the patient could suffer from and which in fact represent the real cause of the fracture, exception making the women at menopause, which make frequent fractures on background of osteoporosis and the cancers with different localizations complicate with bone metastasis, in these situations the fractures appear on pathologic bone and have a reserved prognosis. We should investigate further for other pathologies in order to find the real cause of the fracture. Methods: Present the case of a young girl aged 19, comes for a consultation with her mother with a complaint of fatigue, loss of appetite, lack of concentration and attention at school, getting tired easily after minimal physical and intellectual effort, the patient also mentions that about eight months ago she sustained a minor trauma to her left forearm which resulted in a fracture, which was then followed by a fracture of the radius and resulted in her being referred to the orthopedics department where her arm had to be fitted with metal rods. The principal signs was: bone deformation, bone shortening, thin bones, abnormally fragile bones, small muscles, joints and weak tendons, formation of thick scars, small somatic conformation, defective dentition (incomplete dentition , the teeth was damaged, the teeth falled quickly),the cornea was transparent blue(blue sclera) and also her mother had blue sclera. The molecular genetic test was used for clinic diagnosis confirmation and releaved mutations in COL1 A1 and COL1 A2 genes responsabile of sintesis of type I procollagen and confirmed the disease osteogenesis imperfecta. Results: The case of the fracture of the forearm was a very rare genetic disease - osteogenesis imperfecta- a congenital autosomal dominance. A.D. illness and occur even if only one parent transmits the effected gene. As in this case the daughter inherited the disease from her mother. Conclusions: Initially the case appeared to be a trivial case with a simple forearm bone fracture./Subsequently detailed physical examination revealed clinical signs such as somatic changes dentition and more importantly blue sclera pointing us towards the extremely rare disease osteogenesis imperfecta / An examination of the patient’s mother revealed the same signs, confirming the patient had inherited the autosomal dominant disease from her mother./Starting from a simple fracture and some nonspecific clinical symptoms, the final diagnosis was an unexpected surprise to finally discover a hereditary disease with autosomal dominant transmission which is extremely rare - osteogenesis imperfecta. This clinical case should point out that sometimes when a simple fracture is discovered there may be previously unknown underlying disease which may have contributed to the injury and fractures can also be caused by other bone disease./ The molecular genetic test was used for clinic diagnosis confirmation and releaved mutations in COL1 A1 and COL1 A2 genes responsabile of sintesis of type I procollagen and confirmed the disease osteogenesis imperfecta.

Increasing burden of obesity, the metabolic syndrome (MetS), Type 2 Diabetes Mellitus (T2DM), and CardioVascular Diseases (CVD) in developing countries has created an urgent need for more researches on the various factors responsible for increasing their prevalence. Literature search was carried out using the terms obesity, insulin resistance, diabetes, dyslipidemia, physical inactivity, the metabolic syndrome, and developing countries, in order to review the criteria and risk factors of MetS and its prevalence in Middle East countries from web sites and published documents. The prevalence of the syndrome and its underlying factors was also compared between some of the Middle East and some of European and American countries. The pattern of the components of metabolic syndrome varies in the Middle East region in comparison with Europe and North America. In addition to differences of diet behaviors, the genetic factors and some of geographic dependent elements may influence the variation of metabolic syndrome architecture worldwide particularly in the Middle East region.