Quantitative Analysis of the Proteomic Response of Human Tumor Cell Lines to Sirtinol

My thesis develops and evaluates a novel, hybrid method for mass spectrometry based proteomics, with the goal of increasing the number of protein identifications, while also allowing quantitative analysis to be performed. The hybrid approach I developed uses classical SILAC labeling coupled to a “novel” scheme for enriching cysteic acid containing peptides which are generated by oxidizing the cysteine and cystine residues of proteins. The resultant acidic side chains are used for the selective enrichment of these peptides. As a result, I get the ease of labeling and enhanced quantitative performance of SILAC protocols with the added benefit of the reduction in sample complexity seen with ICAT protocols. The hybrid method enabled me to study the proteomic profiles of HI229 cancer cells following sirtinol treatment. Sirtinol, which targets members of the sirtuin family, has been proposed as a treatment for neurodegeneration and cancer. In fact, sirtinol induces senescence in H1299 cells, and I was able to observe the long term therapeutic effects of sirtinol at the proteome level. The optimized hybrid protocol yielded approximately 5 times the proteins identifications from H1299 cells, when compared to the standard protocol. Quantification of this data revealed that 140 proteins are downregulated and 88 proteins are upregulated following sirtinol treatment of H1299 cells. Moreover, an additional benefit of the protocol was an increase in the total number of post translational modifications (PTMs) identified using the hybrid approach. This work will give a better understanding of the mode of action of these compounds, as well as revealing the biological mechanisms of their effects. In conclusion, the hybrid method is useful for increasing the number of protein identifications from complex samples. It is an accurate and straightforward method for comparative studies. This method is especially useful for drug analysis studies or other binary comparisons

Hypnic headache associated with medication overuse: case report

We have recently evaluated a 54-year-old woman who had migraine without aura in her history but presenting with a typical hypnic headache (HH) which is presumably not a primary headache but associated with an ergotamine overuse headache. Her HH was relieved with a washout protocol which includes 75 mg amitriptyline daily with the addition of metoclopramide and encouraging her not to use any analgesics. Our aim was to report this unique patient to emphasize this rare association and to discuss the possible pathophysiological implications for both of these entities

Nonconvulsive Status Epilepticus

Nonconvulsive status epilepticus (NCSE) has a wide range of clinical presentations, ranging from confusion to coma accompanied by many kinds of continuous or recurrent epileptic discharges on the electroencephalogram (EEG). It may occur not only in epileptic patients but also in adults with no previous history of epilepsy. EEG is the only reliable method of diagnosing NCSE. The identification of NCSE may be particularly difficult and therefore, a high level of suspicion is essential for early diagnosis. In this review, clinical and electrophysiological findings, types, frequency, diagnosis, differential diagnosis, treatment, and prognosis of NCSE are discussed in the light of the relevant literature

Photosensitivity and Reflex Epilepsies

Reflex epilepsies (RE) objectively and consistently provoked by specific external or internal stimuli are important clues for investigating complex mechanisms of epileptogenesis. RE have many intriguing subtypes depending on the trigger. Visually-sensitive epilepsies or photosensitive epilepsies are genetically determined and constitute the major part of RE but their diversity is even huge both clinically and electrophysiologically. Reflex seizures can occur in both focal and generalized epilepsy syndromes and can also be associated with spontaneous seizures. Afferent external stimuli can be: elementary or elaborate, structured and complex. Moreover, triggering activity may also be elementary (motor movement) or elaborate (reading, chess playing), or both (reading aloud). Complex forms of RE pose many different questions and there are reported cases with various symptomatic etiologies indicating some acquired factors besides genetics. This review will focus on the current understanding of RE and its pathogenetic implications

A preliminary proteomic evaluation of smooth muscle cells in thoracic aortic aneurysms

WOS: 000332944300010Aortic aneurysm is characterized as localized degeneration of the aorta leading to advanced weakening and widening of the vessel. While the exact mechanisms have yet to be determined, current studies indicate that the degradation of extracellular matrix (ECM) proteins and apoptosis of vascular smooth muscle cells (SMCs) may result in extendibility, dilation, and rupture of the vessel. Within the aortic wall, SMCs are implicated as key components involved in disease development, as numerous molecular changes have been reported to occur. Most current studies involve either investigation of proteins constituting a group or pathway in SMCs, or analyses of the whole aortic tissue. In order to determine which proteins are important in the development of thoracic aortic aneurysms (TAAs), we performed comparative proteomic analyses using cultured SMCs from TAAs versus controls. Label-free nano LC-MS/MS analysis of cell extracts resulted in the identification of 256 proteins, 26 of which were differentially regulated by >= 1.4-fold. Both previously described and new proteins were identified that were involved in oxidative stress, ECM formation, energy metabolism, or the 14-3-3 pathway. Among these, differential expression of SerpinH1, a protease inhibitor for collagenases, was further verified via immunoblotting. Here we have attempted to shed light on the cellular mechanisms of TAAs.European Union [200647]This work was supported by the European Union 7th Framework Programme Project entitled Fighting Aneurysmal Disease (FAD), Health-2007-2.4.2-2, Grant Agreement No. 200647, and internal funding through TUBITAK.Türkiye Bilimsel ve Teknolojik Araştırma Kurumu (TÜBİTAK

Clobazam as an Add-on Therapy of Patients with Drug-resistant Epilepsy: Experience of a Tertiary Epilepsy Center

Objectives:This retrospective study aimed to overview the efficacy and side effects of clobazam in patients with drug-resistant epilepsy who were followed in a tertiary epilepsy center

HLA-DR and -DQ Associations with Multiple Sclerosis in Turkey

ABSTRACT: The DRB, DQA, and DQB subregions ofthe major histocompatibility complex (MHC) were investigatedby polymerase chain reaction and sequence-specificoligonucleotide probe hybridization (PCR/SSO) in103 multiple sclerosis (MS) patients and 101 healthycontrols from Turkey. Significant differences were detectedbetween MS and control populations in the frequencies ofDRB1*1501 [29 vs. 14, p 5 0.02, odds ratio (OR) 5 2.4],DRB1*04 (35 vs. 18, p 5 0.01, OR 5 2.3), DQB1*0302(30 vs. 15, p 5 0.02, OR 5 2.3), DQB1*0602 (27 vs. 10,p 5 0.005, OR 5 3.2), DQB1*0501 (10 vs. 24, p 5 0.01,OR 5 0.3), DQA1*0101 (16 vs. 31, p 5 0.02, OR 50.4), and DQA1*0103 (7 vs. 19, p 5 0.02, OR 5 0.3).These results confirm the proposed positive association ofthe Dw2 (DRB1*1501 DQA1*0102 DQB1*0602) haplotypewith MS in Caucasians in our Turkish population(25 vs. 8, p 5 0.003, OR 5 3.7). Furthermore, the“putative” haplotype supposed to be more frequent in theMS population of Mediterranean countries, namelyDRB1*04 DQA1*03 DQB1*0302, is also associatedwith MS in Turkey (29 vs. 12, p 5 0.006, OR 5 2.9).The presence of two different haplotypic associations inMS emphasizes the complexity of the genetic susceptibilityto MS in different populations