Qatar Foundation Annual Research Forum Volume 2013 Issue 1

One important type of speech sound disorders is articulation and phonological disorders that affect children of age between 2-8 years. All normally developing children follow a unique developmental timetable, table 1 in which they acquire articulation and phonological skills to correctly pronounce both consonants and vowels to make the sound patterns of their respective language. Previous statistics, table 2 show that the number of Arab children, in the age group 2-8 years, who suffer from articulation and phonological disorders, is significant. To be able to test, assess, and evaluate the effect of therapy on these disorders, there is need to conduct research and development that would culminate in the availability of speedy, versatile and interesting computerized tools for use by Speech and Language Therapist, SLPs in standardized test, assessment, and evaluation of articulation and phonology disorders among the children, figure 1. There are prerequisites to the development of these tools such as the development of a suitable phonetic and phonological vocabulary (see table 3) of Modern Standard Arabic, MSA that is spoken by such children and which will meet the test and assessment requirements for speech disorders. The vocabulary could then be used to create a speech corpus for children with normal speech who are in the same age group as the children with disordered speech. Such speech corpus could then be used as a frame of reference to set the standards and norms for children speech. The standards and norms can be used by SLPs to compare normal children against children suspected of having the disorder. The speech corpus could also be used by researchers to develop limited vocabulary, speaker-independent phonetic speech recognizers that would help SLPs during speech therapy and evaluation. To meet the ultimate goal of finally producing efficient and versatile computerized tools to assist SLPs in their mundane tasks of handling articulation and phonological deficits among children in an efficient manner, we envisage a project with the following objectives in mind: 1) develop a comprehensive articulation and phonology vocabulary that represent all the sounds of MSA spoken by the children in all their important articulation and phonological contexts, and 2) use the vocabulary to create a norm speech corpus that represent the articulation of all sounds for children with healthy speech production. The corpus should include significant quantities of speaker-dependent data for a significant number of children with normal speech and would be segmented, labeled and transcribed according to the standards used for such tasks. The development of test and assessment vocabulary and the creation of the speech corpus from such vocabulary are the prerequisites for any investigation on articulation and phonology disorders. Firstly they would help the research community to study the disorders and secondly, after the development of suitable computerized tools, they assist the SLPs to test, assess, and evaluate the effect of treatment for articulation and phonological disorders among children. In this article, we present the status of the research and outline a roadmap to achieve our objectives and ultimate goal.

Autism spectrum disorders (ASD) are a group of neuro-developmental disabilities, which, in many cases, is characterized by severe impairments in communication skills, social interaction, psychomotor coordination, behavioral control and emotional stability often accompanied by deficits in cognition, attention and sensory processing. According to a recent study conducted by Dr. Fouad Alshaban (Shafallah Medical Genetics Center, Doha, Qatar), the accurate prevalence rate of autism-spectrum disorders in Qatar is still uncertain. However, there is a general concern that it might be increasing in the last few years. The symptoms of ASDs are typically present before age 3 years, but frequently a formal diagnosis take longer to be established, which may compromises the efficacy of developmental and life-quality improving strategies. In most countries of the world the scenario is not much different. It has been shown that autism-associated neuroligin-3 mutations disrupt tonic endocannabinoid signaling in animals. Endocaanabinoids are neuromodulators produced by the brain, which act upon the same receptors and neuronal circuits affected by exogenous cannabinoids from the plant Cannabis sativa. This system is central for brain homeostatic activity control, learning, social interaction, memory formation and emotional modulation, among several neuronal and physiological functions. Here we present our proposal to use primates and valproate-treated rodents (an animal model for autism) in order to deepen our knowledge about the involvement of defects in endocannabinoid system as etiological factors capable of generating behavioral and cognitive aspects related to human autism. The reasoning behind the use of two species is to facilitate the conceptual bridge between the findings obtained in valproate-treated rodents and the understanding of the importance of the endocannabinoid system for social interaction and emotional behavior in primates. At first, this combination has the potential to point out possible pharmacological treatments based on modulation of the endocannabinoid system and in the identification of biomarkers in rodents. Secondly, this knowledge can help to develop, and to ethically justify, the creation of primate models for the study of pharmacologically treatable aspects related to human behavior that cannot be studied in rodents. In Brazil we have the privilege to work with non-human primates in a high-standard, naturalistic facility, the Primatology Center of the University of Brasilia (in the capital of Brazil). Therefore, we are taking advantage of this capability to test new hypotheses relating endocannabinoid signaling defects with autism symptoms and eventually generate a non-human primate animal model that, in one hand, may help on the discovery of molecular markers for early diagnosis and, on the other hand, may help the development of pharmacological tools to treat important symptoms of ASD. We want to share and discuss our ongoing project in this meeting as part of an effort to foster found raising and potential collaborations between our group and research groups from Qatar.

The interest in bone and softtissue replacement achieves rapid advances throughout the last decades. Injured bone has a unique ability to regenerate which leads to complete anatomic and physiologic repair. However, not every bony defect can beself-repaired. Repair of large osseous defects caused by neoplasms, cysts, trauma, infection, congenital diseases and surgical intervention are major problems in oral and maxillofacial field. The current approach is to use bone and bone substitutes for filling such defects. The aims of filling such osseous defects are to preserve the morphologic contour, restoration of the mechanical strength and function, elimination of the dead space to reduce postoperative infection, and the prevention of ingrowths of soft tissue. When conditions are favorable for regeneration of bone, such goals can be achieved without the need for grafting techniques. However, in many large lesions or in the absence of favorable conditions in small defects, the morphological contour is not completely restored and the bone remains weak. Different grafting materials have been introduced into the field in the last decade. The profound limitations of biological grafts have prompted the use of synthetic materials as an alternative to autogenous and allogenic bone grafts.In the present invention, we succeeded in manufacturing of a novel biomaterial in the field of bone healing. Our novel biomaterial synthesized from bovine serumalbumin using our newly developed technology (sub-critical water technology). The biomaterial showed a variety of mechanical properties and biodegradability that allowed for its application in the biomedical field and in particular as an alloplastic bone substitute. In our way to validate the applicability of our novel biomaterial, we co-operated with the department of Oral and Maxillofacial Surgery Faculty of Dentistry, Al Minia University, to evaluate our material as an alloplastic bone substitute through animal test. The results revealed satisfactory results when used as a bone grafting material in surgically created bone defects in the rabbit models. The obtained results showed that these novel materials have the following properties: Biocompatibility, Nontoxic and nonirritant, Act as a haemostatic agent and maintained the blood clot inside the defect, Osteoconductive property, Biodegradability property,easily manipulated and handled.The most important findings from the obtained results revealed that these novel materials have the ability to accelerate bone healing specially at the early stages following implementation of the material. Part of the work was patented in Japan with patent number 2005-028066 and published in high impact factor scientific journals worldwide 1, 2,3, 4, 5 . In the presnt work the most new obtained data concerning biocompatibility and animal test will be presented

The sensitivity of magnetic resonance (MR) to diffusion has long been exploited as a method for investigating molecular motions in complex systems. The recognition that MR can be made sensitive to the details of anisotropic diffusion in structured materials led to the formulation of the problem as a tensor (DTI) rather than a scalar. New directions exploiting MR diffusion imaging techniques can yield essential new information previously not accessible with other imaging modalities as potential cancer biomarkers. We investigated the feasibility of applying DTI for in-vivo fiber tractography of the prostate. We carried out retrospective study of men with biopsy proven prostate cancer. All patients had undergone prostate MRI with an endorectal coil on a 1.5 T MRI scanner. Diffusion weighted images were acquired with a single shot echo-planar acquisition, with six diffusion sensitizing directions. Preliminary fiber track data was generated as in Fig. 1 using Diffusion Toolkit and displayed with TrackVis (trackvis.org). This data was then used to identify multiple hand drawn regions of interest (ROI) over areas of pathologically proven tumor, the central gland and the normal peripheral zone of the gland. For quantitative analysis, we introduced a track density parameter, which is the number of tracks in a given ROI divided by its volume, as a normalized measure of the tracks passing through the ROI. The values were statistically analyzed and correlated with the staging. One of the key goals of the study is to apply this method to analyze post radiation treatment cases where other modalities fail to yield contrast necessary to distinguish cancerous tissue from necrosis. The main challenge of fiber tractography, however, is the existence of multiple-fiber orientations within an imaging voxel which renders the diffusion tensor model inadequate. The practical limitations imposed by the requirement for a wide spectral sampling of the diffusion spectrum in q-space, can be met if these impulse diffusion gradients are replaced with chirped oscillatory gradients. In this abstract, asymmetrical chirped diffusion sensitizing gradients are shown to yield an efficient sampling of q space in a manner that asymptotically approaches the spectral coverage afforded by delta function diffusion sensitizing gradients. The asymmetrical nature of the gradients is an extension of the method recently proposed by Laun et al.[1] that allows the diffusion experiment to preserve the phase information and hence reconstruct the exact shape of the underlying structural restrictions -see Fig. 2. The challenge is the consequent reduction in diffusion sensitivity as one probes higher frequency dynamics. This problem is addressed by restricting the gradient power to a spectral bandwidth corresponding to the diffusion spectral range of the underlying restrictive geometry. Simultaneous imaging of diffusion at microscopic resolution and at temporally resolvable diffusion time-scales thus becomes possible in-vivo. Fig. 2: Inverse imaging of a triangular restriction using asymmetrical diffusion encoding gradients [1] F. B. Laun, T. A. Kuder, W. Semmler and B. Stieltjes, Phys. Rev. Lett. 107, 048012 (2011).

Autism spectrum disorder (ASD) is a clinically and etiologically heterogeneous condition with high prevalence amongst all world populations. Due to the life long morbidity, ASD poses a profound world-wide public health challenge. Despite its obviously high heritability, a genetic cause is only identified in a small fraction of patients due to its complex etiologic heterogeneity. Recently, whole genome sequencing (WGS) emerged as a powerful tool for variant discovery due to its comprehensive and uniform coverage and is expected to be beneficial for studying ASD. We used WGS to examine 20 families with ASD, mainly from Qatar, as well as from a few other Arabic countries, to detect de novo or rare inherited genetic variants predicted to be of pathogenic significance. All probands were diagnosed with ASD by at least an ADI-R (Autism Diagnostic Interview-revised) evaluation, in addition to other screening or confirmatory tools and were recruited from the Shafallah Center for Children with Special Needs. Due to the nature of the ascertainment source, all probands had associated intellectual impairment. In all probands known causes of ASD were excluded, fragile X syndrome by PCR and Southern blott, Rett syndrome (for females) by Sanger resequencing and MLPA and Copy Number Variations (CNV) by SNP genotyping on an Illumina 1M-Duo SNP chips. Among all probands, we identified deleterious de novo mutations in six (30%) families and inherited X-linked in two families (10%). We did not identify any inherited autosomal dominant mutations, but identified putative inherited autosomal recessive varations in three families (15%). The deleterious de novo variants were found in five previously unrecognized autosomal genes, and in one known autosomal ASD gene. The inherted X-linked variations were found in ASD risk genes. We are interested in deeper examination of the putative variations with autosomal recessive pattern of inheritance, since the parents of 60% of the probands are consanguineous. The deeper examination will employ the study of knock-out and knock-in mouse models and different in silico and in vitro protein modeling studies. These results so far suggest that WGS of a trio coupled with thorough bioinformatic analyses provide a powerful diagnostic tool in ASD. It is also a potent methodology for gene discovery in ASD, which may provide the basis for genetic testing for early diagnosis and early targeted intervention with a final goal of better outcome.

Exome sequencing was used to study genetic variation in the Qatari population. We sequenced 100 healthy Qataris (50X average depth, with >80% of sites at >10x depth) representing the 3 major Qatari genetic subpopulations (Q1 - Bedouin, Q2 - Persian/South Asian, Q3 - African). A total of ~174,000 high quality variants were identified, of which 1306 were predicted to cause loss of function (frameshift/stop-gain/splice/start-loss) in 1102 unique genes. Of these, 471 (36%) were novel Qatari variants (absent from public databases) of which 43 were observed in 2 or more individuals, suggesting a population allele frequency =1%. 54 of 471 loss of function mutations affected genes in the OMIM database with a confirmed disease-causing status. Of these, 30 occurred in genes known to cause severe autosomal recessive disease. Of interest, some of these mutation had allele frequencies of up to 8% in Qataris. We further focused on the subset that were nonsense mutations (n=671/1306). These occurred in 616 of 1102 unique genes and resulted in a median truncated protein length of 54.3%. 237 of 671 nonsense mutations were novel to Qataris, suggesting that a significant number of variants from this population are yet to be sampled and covered in public databases. When nonsense alleles were considered in the 3 separate Qatari subpopulations, the Q3 population displayed the highest number of nonsense alleles per individual (mean: 40.7, range: 28-55), consistent with the highest genome diversity due to African ancestry. Surprisingly, despite having the lowest number of heterozygous nonsense mutations (mean: 22.8, range: 16-31), the Q1 subpopulation had at least 20% more homozygous nonsense mutations (range: 2-15, mean: 6.7) per individual than either Q2 or Q3, consistent with the high levels of consanguinity in this population. Specifically, the Q1 had significantly higher allele frequencies than the rest of the world for 2 autosomal recessive disease genes, including 1 individual homozygous for a nonsense mutation in POMT1 which causes muscular dystrophy, and 4 individuals heterozygous for a mutation that truncates the Holoprocencephaly-causing TGIF1 gene to 7.5% full length. Additionally, the population as a whole had higher allele frequencies for 6 other autosomal-recessive-disease-causing mutations, bearing significant health implications for the Qatari population in particular, and for this highly consanguineous region of the world in general.

[Background] Chlamydia trachomatis (CT) is one of the most common bacterial sexually transmitted infections (STIs) worldwide, and is a major cause of pelvic inflammatory disease, infertility, and ectopic pregnancy. A recent study has found larger than expected prevalence of CT in Qatar, with a prevalence of about 5% among women attending primary health-care centers. The driver of such higher than expected prevalence is not clear. We examined whether the lack of screening programs for CT could contribute to explaining this level of prevalence. [Method] We constructed a mathematical model to examine the public health impact of different CT treatment approaches. The model was parameterized by the prevalence of CT in Qatar along with state of the art empirical evidence of CT natural history and transmission. The population was divided into different risk and age groups, and a mixing matrix was introduced to describe the mixing between these groups. The impact of treatment was assessed at endemic equilibrium. Univariate sensitivity analyses were conducted. [Results] Nearly 63% of CT infections in the population were asymptomatic. A symptomatic treatment approach, where 90% of symptomatic cases were treated, reduced CT prevalence by only 6.5%. An opportunistic screening program, where 20% of the population was screened annually for CT, reduced the prevalence by 36%. Routine screening program, where 50% of the population was screened annually for CT, reduced the prevalence by 93%. The large impact of screening on CT transmission was driven not only by the diagnosis of asymptomatic infections and treating them, but also by reducing effectively the duration of the infectious period during which infected persons can transmit the infection to other individuals. [Discussion] Our results suggest that a key contributor to the higher than expected prevalence of CT in Qatar, is the non-availability of CT-specific screening programs to detect and treat asymptomatic CT infection, such as those available in West Europe and North America. Current programs that focus on treatment of syndromic cases are missing the majority of cases. The undiagnosed infections are driving further transmissions, and a larger CT prevalence. Opportunistic targeted screening could be a meaningful starting point for CT screening programs in Qatar, with potentially routine Pap smear testing as an entry point. Our findings indicate the utility of developing STI-specific programs in Qatar as part of the large expansion of health care services in this country. Detecting CT infections and linking them with appropriate treatment channels could alleviate an unnecessary disease burden and its consequences in the population.

Acute respiratory failure is one of the major causes for concern in intensive care units (ICU). It is a condition that occurs when fluid builds up in the air sacs in the lungs. When this happens the lungs cannot release oxygen into the blood and get rid of carbon dioxide from the body. The lack of oxygen in the blood will cause the organs to malfunction and in severe cases could lead to failure of vital organs such as heart and brain. Patients with acute respiratory failure are usually supported by the mechanical ventilator. The goal of mechanical ventilation is to ensure adequate ventilation, which involves a magnitude of gas exchange that leads to the desired blood level of carbon dioxide, and adequate oxygenation, which involves a blood concentration of oxygen that will ensure proper function of vital organs. A plausible reference lung volume pattern, which corresponds to an optimal patient outcome, is specified by clinical personnel and the task of automated mechanical ventilation is to apply appropriate input pressure so that the output from the system is able to track the given reference voulme pattern. One of the main challenges in mechanical ventilation that needs to be carefully considered is that the applied pressure cannot be arbitrarily large since large applied pressure could potentially damage the lungs, and hence, worsen patients outcome. Another challenge in designing an efficient control algorithm for mechanical ventilation is that the parameters characterizing the dynamics of the lungs, that is, the lung resistances and lung compliances, vary from patient to patient as well as within the same patient under different conditions. Furthermore, the volume of each compartment of the lungs cannot be directly measured and only the total volume of the lungs is available. Therefore, a robust control methodology, which only uses output information and limited applied pressure, is needed in order to design an efficient control algorithm for automated mechanical ventilation. In this research work, we propose an output-feedback sliding mode control methodology for a nonlinear multicompartment respiratory system with amplitude and integral input constraints. The amplitude input constraint is needed to ensure that the applied pressure is not too large, as not to damage the lungs, while the integral input constraint enforces the upper bound on the amount of work done by the ventilator. The proposed controller only uses output information (i.e., the total volume of the lungs) and automatically adjusts the applied input pressure so that the system is able to track a given reference volume pattern in the presence of parameter uncertainty (i.e., modeling uncertainty of the lung resistances and lung compliances) and system disturbances. A Lyapunov-based approach is presented for the stability analysis of the system and the proposed control framework is applied to a two compartment lung model to show the efficacy of the proposed control method.

Background: Myeloproliferative Neoplasms (MPNs) are clonal hematopoietic disorders, classified into three different phenotypes Polycythemia Vera (PV), Essential Thrombocytosis (ET) and Primary Myelofibrosis (PMF). This disorder is associated with the presence Jak2V617F mutation in 90% of PV and 50% of ET and PMF patients as well as other mutations such as Jak2 exon 12 and MPL. However, there are still significant numbers of MPNs cases that are negative to most common genetic anomalies and many mutations are still unknown Aim: To identify and elucidate genetic defects causing MPNs that could serve as disease biomarker. Methods: 7 MPN patients and healthy individuals from 3 consanguineous families were consented into the study. Peripheral blood samples were collected from 6 patients and 4 healthy individuals. Further, genomic DNA was extracted and used for multiplex PCR amplification of 190 amplicons targeting 739 cancer associated mutations in 604 loci from 46 key cancer genes, using the Ion AmpliSeq™ Kit. Next Generation Sequencing (NGS) was performed through the Ion Torrent Personal Genome Machine using the 318 chip and was analyzed with the Torrent Suite Software (Full list of mutations targeted can be found at http://www.bcm.edu/geneticlabs/index.cfm?pmid=21681). Mutation details were obtained from the Catalogue Of Somatic Mutations In Cancer (COSMIC) database. A human genome (hg) 19 DNA sequence (http://www.ncbi.nlm.nih.gov/refseq/rsg/) was used as references for this panel of genes. The nomenclature of mutations is based on the convention recommended by the Human Genome Variation Society (http://www.hgvs.org/mutnomen/). The confirmation of NGS data was performed using RQ-PCR or Sanger sequencing and Jak2 exon 12 mutations were studied. Results: Out of four PV patients, three were positive for the JAK2 V617F mutation. One patient had mutations in PDGFRA 838-840delGLA, APC Q1285fs, NPM1 Splice Site Loss and PTEN S10N. The second patient had mutations in KIT M541L, KDR Q472H, TP53 P72R, while the third patient had PI3KCA I391M, KIT M541L, KDR Q472H and P53 P72R mutations. The fourth patient was negative to the JAK2 V617F mutation, but showed PDGFR A838-840delGLA and SMARCB1 T72K mutations. Out of three ET patients, one patient was positive for JAK2 V617F, SMARCB1 T72K, IDH1 K115Q and PDGFRA 838-840delGLA mutations. Two patients were negative for the JAK2 V617F and MPL mutations, one patient had mutations in PDGFRA 838-840delGLA, APC Q1285fs and SMAD4 198-200delPAL, while the other patient had mutations in CDKN2A S73R, APC Q1285fs and PDGFRA 838-840delGLA. Furthermore, the PDGFRA 838-840delGLA and APC Q1285fs mutations were also found in healthy individuals. Conclusion: This study was able to identify a list of deleterious somatic mutations such as missense and frame shift mutations, in-frame deletions, splice loss sites and Single Nucleotide Variation (SNV) in MPNs patients and healthy individuals. Our preliminary results suggest that the MPNs patients in Qatar have complex mutations. Evidences show that there exists a possibility of the disease arising out of the accumulation of genetic alteration and not as the consequence of a single genetic event. This could possibly be due to the high rate of consanguineous marriages in Qatar i.e. the "Founder Effect".

Calcineurin-NFAT Signaling Pathway Mediates NHE1 Induced Cardiac Hypertrophy Mohamed Mlih, Fatima Mraiche College of Pharmacy, Qatar University, Doha, Qatar In the myocardium, the Na+/H+ exchanger isoform 1 (NHE1), a plasma membrane protein that regulates intracellular pH has been shown to be critical in the development of cardiac hypertrophy. Inhibition of NHE1 activity/expression has previously been demonstrated to produce cardioprotective effects. Recent reports have reported that transgenic mice overexpressing active NHE1 developed spontaneous cardiac hypertrophy in association with elevated levels of osteopontin (OPN). Osteopontin is a secreted protein involved in many physiological pathways including bone formation, immune response, vascularization and cardiac hypertrophy. The physiological pathway in which active NHE1 induces OPN expression in the heart remains unknown. Recently, NHE1 was described as a key regulator of the calcineurin-NFAT pathway. The calcineurin-NFAT pathway is a hypertrophic pathway induced by intracellular increase in calcium concentration that induces NFAT translocation to the nucleus. In the nucleus, NFAT interacts with the transcription factor GATA-4 and together activate hypertrophic gene. Our hypothesis is that NHE1 through the calcineurin/Nfat pathway induces OPN expression leading to cardiac hypertrophy. To verify this hypothesis , rat cardiomyoblasts (H9c2 cells) were infected with activate NHE1 adenovirus in the presence and absence of a calcineurin inhibitor, FK506 . H9c2 cells infected with active NHE1 showed an increase of NHE1 activity (NHE1: 376.5%) and exhibit a 1.6 fold increase in cell area compared to the control, which was attenuated in the presence of FK506. Our preliminary results show that NHE1 activation increases osteopontin protein expression. In addition, H9c2 cells infected with active NHE1 also demonstrate a significant activation of the transcription factor GATA-4 (NHE1: 149% ±28% , p<0.05). The activation of GATA-4 induced by active NHE1 is inhibited by FK506 treatment. In conclusion, NHE1 through the calcineurin/NFAT pathway induces cardiac hypertrophy in H9c2 cells. This new finding will give us a better understanding of signaling pathway mediating NHE1 induced cardiac hypertrophy and allow us to identify alternative therapeutic targets for the treatment heart failure.

Various betaine materials - single molecules or polymers - represent one of the best tools for prevention of biofouling on surfaces due to their biomimetic character and unique properties [1]. Betaines consist of internal salts in a single unit between positive quaternary ammonium and negative sulfo, carboxy or phosphate groups. This contribution highlights our progress in preparation and application of betaine materials to resist non-specific interactions on a surface of biosensors and membranes. Various betaine derivates with sulfo- [2] or carboxylbetaine groups were synthetized from lipoic acid as a natural precursor. Disulfide moiety from lipoic acid structure assists in attachment of a betaine derivative to a gold surface applied for preparation of biosensors. Biosensor devices were characterized by a set of tools including electrochemical impedance spectroscopy, FTIR, AFM, XPS, QCM, etc. Betaine derivatives introduced to the surface of various biosensors allowed their effective work in complex samples such as human plasma with a high reliability of detection and with a detection limit down to a femtomolar level for detection of specific glycoproteins. Carboxybetaine derivate provided a dual function for preparing biosensors i.e. a non biofouling surface and introduction of functional groups for covalent immobilization of sensoric biomolecules after performing a NHS/EDC coupling chemistry, as well. Moreover, formation, characterization and application of nanofibres and hydrogel from sulfo and carboxybetaine polymer as a membrane precursor will be discussed. Optimal formation of nanofibres was performed via an electrospinning process from trifluoroethanol solution. Characterization of membranes prepared was done by water sorption, FTIR, AFM and SEM techniques. Preliminarily proteins and cells attachment tests showed a dramatic decrease in adsorption and adhesion. Keywords: betaines, biomimetic, biosensor Acknowledgement: The financial support from the Slovak research and development agency APVV 0282-11 is acknowledged. The research leading to these results has received funding from the European Research Council under the European Union's Seventh Framework Programme (FP/2007-2013)/ERC Grant Agreement No. 311532 References: [1] P. Sobolciak, I. Lacik, P. Kasak Chem. Papers, 2011, 105, 918-925. [2] T. Bertok, L. Klukova, A. Sediva, P. Kasak, V. Semak, M. Micusik, M. Omastova, L. Chovanová, M. Vlcek, R. Imrich, A. Vikartovska, J. Tkac Anal. Chemistry 2013, 85, 7324-7332.

Background: The efficacy of medical male circumcision (MMC) in reducing female to male HIV transmission has been established by three clinical trials. Programs are being developed for massive roll-out of MMC as an HIV intervention across multiple countries in sub-Saharan Africa (SSA). We aimed in this study to explore generic results of the effectiveness of MMC as an HIV intervention. Method: A population-level deterministic model was constructed to describe a prototype heterosexual HIV epidemic in SSA. The model consists of a set of coupled nonlinear differential equations that stratifies the population into compartments according to sex, circumcision status, age group, sexual risk group, and HIV status and stage of infection. The model was parameterized by state of the art empirical data of HIV natural history and transmission. Sensitivity analyses with respect to different effects were conducted. Results: The effectiveness of MMC, defined as the number of MMCs needed to avert one HIV infection, was inversely proportional to HIV prevalence with as little as about 10 MMCs needed for each infection averted in settings at high HIV prevalence. The effectiveness also varied substantially with each of the 5-years age groups targeted, with the highest effectiveness found by targeting the 20-24 years age group. Risk targeting strongly affected the estimated effectiveness. MMC effectiveness was an order of magnitude higher by targeting the high-risk groups as opposed to the general population. The speed of scale-up influenced also the estimated effectiveness in the short-term, but was less influential for long-term time horizons. MMC was found to be generally cost-effective, with the cost per infection averted being less than $1000 for most settings. Conclusion: Our findings demonstrate that MMC is an effective HIV prevention intervention, and likely to be cost-effective in most settings in SSA. The highest MMC effectiveness is attained by targeting young sexually active males and high-risk groups. These generic results of MMC effectiveness suggest the need for custom-designed assessments for each country separately, taking into account the likely temporal evolution of the epidemic in the next few decades, the age and geographic distribution of HIV infection, population pyramid, and the country-specific cost for each MMC performed.

Back ground The volume of cardiac diagnostic procedures involving the use of ionizing radiation has increased rapidly in recent years. Coronary computed tomography angiography (CCTA) has been increasingly used in the diagnosis of coronary artery disease (CAD). The CT is an excellent technique to make a diagnosis of disorders in many different parts of the body, however, because the heart is constantly beating, CT scan could never be used to image it in the past. The main advantages of coronary CT is non-invasiveness, rapid acquisition of high-resolution images and high diagnostic accuracy .With rapid improvements in spatial and temporal resolution, CCTA not only visualizes coronary anatomy and characterizes plaque components, but also allows for quantitative analysis of coronary stenosis. OBJECTIVE. The purpose of this study was to assess and compare the radiation doses of different coronary CTA (CTA) protocols: second-generation dual-source 128-MDCT, and single-source 64-MDCT. MATERIALS AND METHODS. CT Systems and Protocols Coronary CTA protocols used with two different systems were evaluated: a single-source 64-MDCT scanner with adaptive section collimation (Somatom Definition 64 AS), and a second-generation dual-source 128-MDCT scanner (Somatom Definition Flash). DISCUSSIONS This study highlights two important findings in the radiation dose associated with the prospective ECG-triggered CCTA. Firstly, a low radiation dose can be achieved in CCTA between different generations of CT scanners with the application of the prospective ECG-triggering protocol. Secondly, BMI affects the radiation dose significantly. RESULTS. Regardless of coronary CTA protocol and CT system, imaging at 100 kV lowered the ED 40-50%. In retrospectively gated 120-kV coronary CTA, the ED ranged from 4.4-21 mSv and in prospectively triggered 120-kV step and shoot coronary CTA, the ED ranged from 3.08- to 7.8 mSv. The lowest ED of all protocols (1.2 mSv) was observed in prospectively triggered high-pitch 100-kV coronary CTA performed with dual-source 128-MDCT. Patient measurements showed similar dose reductions for prospective triggering and low voltage settings without an influence on signal-to-noise ratio or image quality. CONCLUSION. In conclusion, a low radiation dose can be achieved in a low and regular heart rate with a prospective ECG-triggering protocol, regardless of the CT scanner generation. Although there is no significant difference in the effective dose between genders, BMI is identified as the main factor that significantly affects the radiation dose in prospective ECG-triggered CCTA in this study. A combination of prospective triggering with low voltage settings and high pitch is an effective measure for reducing the ED of coronary CTA to values of 1.2-4.8 mSv. it offers a dose reduction potential of more than 80%, which to our knowledge has not been achieved with any other coronary CTA technique. the High-pitch helical coronary CTA is associated with a very low radiation dose is one of the good option for any general coronary screening with the limitation of single phase images and the prospective sequencial mode is almost (95%) suit for all patients with bit more radiation dose than high pitch with the advantage of multiphase and diagnostic accuracy.

Heart failure (HF) is one of the most common causes for death, to prevent HF the understanding of the (patho-)physiology is key and the development of efficient tools is an important contribution for cardiac research. Cardiac tissue slice is an increasingly popular model for cardiac electrophysiology research and pharmacological compound testing. We use optical mapping to investigate cardiac tissue slices. In this study, we perform dual imaging of trans-membrane potential (Vm) and the intracellular free calcium concentration transient (CaT) in tissue slices from the rabbit, using a relatively high spatio-temporal resolution. We detail our method for processing of the data to extract relevant characteristics of the action potential (AP) and CaT. We found that slices needed a recovery time of about 40-70 minutes after cutting, before the AP reaches a steady-state. To characterise the CaT, AP and conduction properties, we used a combination of multi-point and field stimulation, to avoid results biased by source-sink mismatches. The tight control of experimental conditions (e.g. slice 'recovery protocol' and stimulation method), leads to reproducible results, this was shown in our other studies based on using this standardised methodology [1,2]. The monitoring of multiple parameters (Vm, CaT) simultaneously at high spatial resolution (compared to patch clamp, sharp electrode, or multi-electrode arrays) allows one to study not only the properties of each parameter in a spatially detailed manner, but also the spatio-temporal interrelation between them. Especially for drug safety studies this method is superior to lower resolution methods. References: 1. Lee P, Klos M, Bollensdorff C, Hou L, Ewart P, Kamp TJ, Zhang J, Bizy A, Guerrero-Serna G, Kohl P, Jalife J, Herron TJ (2012) Simultaneous Voltage and Calcium Mapping of Genetically Purified Human Induced Pluripotent Stem Cell-Derived Cardiac Myocyte Monolayers. Circ Res 2. Lee P, Wang K, Woods CE, Yan P, Kohl P, Ewart P, Loew LM, Terrar DA, Bollensdorff C (2012) Cardiac electrophysiological imaging systems scalable for high-throughput drug testing. Pflugers Arch 464 (6):645-656. doi:10.1007/s00424-012-1149-0

Cystic Fibrosis (CF) is a genetic disorder that is caused by mutations in the gene for the CF Transmembrane Conductance Regulator (CFTR). CFTR is an ABC transporter chloride channel containing five domains: two membrane-spanning domains (MSD1 & 2) connected by two nucleotide-binding domains (NBD1 & 2) and linked by a regulatory domain. The most common CF causing mutation is the deletion of phenylalanine 508 (?F508) in NBD1. It was shown that ?F508 thermodynamically destabilizes NBD1, as a result, misfold and degrade CFTR channel in the endoplasmic reticulum and prevents its processing and translocation to the plasma membrane. An effective drug that rescue the channel enhance its folding will increase its concentration on the plasma membrane. Here, we show stabilization of ?F508-NBD1 with second site mutations was not sufficient to increase the ?F508-CFTR folding efficiency and membrane concentration to that of the wild-type level. However, the introduction of additional mutations that stabilizes the interaction between NBD1 and MSD2 of CFTR in the presence of ?F508-NBD1 stabilization mutations increased the ?F508-CFTR folding efficiency and biogenesis from ~2% to 80% of the wild-type. As a result, a two-component drug that energetically stabilizes ?F508-NBD1 and maintain the NBD1-MSD2 interface interactions are required for wild-type like folding, processing, and transport function, suggesting a two step correction process.

Qatar Biobank (QBB), the first very large scale, long-term public biorepository in Qatar, is designed to build a powerful research infrastructure for future investigations of the lifestyle, metabolic and genetic risk factors by collecting comprehensive phenotypic baseline data among healthy volunteers, including ECG, blood pressure, anthropometry, spirometry, retinal imaging, carotid 3D ultrasound, arterial stiffness, total body iDXA, in addition to detailed personal lifestyle and clinical data. QBB collects and stores blood samples subdivided in 68 aliquotes for different future research purposes. Qatar Biobank understands that building a successful biobank depends on the willing participation of the public to come forward to contribute. The recruitment of participants requires insight into the public's existing knowledge of biobanking, level of willingness and an understanding of the motivators and barriers to participation. From December 2012 to June 2013, 503 participants completed anonymously a feedback form to evaluate their experience. The aim of this specific survey is to gain insight into recruitment methods, incentive to participate and satisfaction with various aspects of the visit. This will enable assessment of the processes of registration, scheduling, checking in, consent and reporting back results. Around 75% of those who completed the feedback form participated in QBB to improve the health of future generations and to have a comprehensive health checkup. 95% were willing to participate again if needed and 85% would recommend participation to friends and family. 87% and 95% thought the length of the questionnaire was appropriate and the questions not too intrusive, respectively. Finally, 91% found the staff welcoming, 89% rated the check-in process as "good" and 76% found scheduling an appointment straightforward. Further comments from participants suggested appreciation of the protection of confidentiality and privacy that was displayed throughout the process and that the process was smooth and enjoyable.

Purpose: The main objective of this study was to develop and validate a simple, rapid and stability-indicating ultra-performance liquid chromatography (UPLC) method for the determination of Letrozole in pharmaceutical gels. Methods: Chromatographic separation was achieved using Aquity UPLC BEH C-18 (2.1 x 50 mm, 1.7µm particle size) with an isocratic mobile phase consisting of acetonitrile: water (35:65, v/v). The flow rate was set to 0.3 ml/min, injection volume was set to 1µl, and the UV detection was carried out at λ= 240 nm. The method was validated as per ICH guidelines. Letrozole was subjected to different stress conditions (acidic, basic, oxidative, thermal, UV-radiation) to assess the stability indicating power of the developed UPLC method. Letrozole was also analyzed in different emulgel formulations containing a mixture of oil, surfactants, co-surfactants and gelling polymers. Furthermore, the stability of letrozole was assessed in the presence of different emulgel ingredients , where the mixtures of the saturated solubility studies were kept at 50°C, and then analyzed by the proposed method at predetermined time intervals. Results: The retention time of Letrozole was 1.80 min (RSD=0.12%). The method was linear over the concentration range of 2.5-200 µg/ml (R2 = 0.9999). The limit of detection (LOD) and the limit of quantitation (LOQ) were found to be 0.025 µg/ml and 0.125 µg/ml, respectively. The inter-day and intra-day precisions (% RSD) were less than 2%. All validation parameters were in the acceptable range. Neither the degradation products, nor the emulgel components interfere with the analysis of letrozole peak. Percentage recovery in emulgels were in agreement with the labeled spiked amounts. Letrozole was found unstable in the presence of PEG400, diethylene gycolmonoethyl ether, oleic acid, glycerylmonooleate when stored at high temperature of 50°C. Conclusions: The proposed method is simple, rapid, accurate, and can be successfully used for the analysis of letrozole's content and its stability in the pharmaceutical emulgels. Acknowledgements: This work has been financially supported by a grant to HM Younes and AA Sallam provided by Qatar National Research Foundation through the National Priorities Research Program grant # 4 - 496 - 3 - 157. Ahmed Abu Helwa is a research assistant financially supported by the same grant.

Cardiovascular diseases are a major cause of global disability and death worldwide. The prevalence of heart disease is quite high in the Unites States and similar trends are becoming apparent in various Middle Eastern and Gulf region countries such as Qatar. It has been suggested that within 10 years, atherosclerosis and its complications, mainly coronary heart disease, will become the leading cause of death and loss of productive life years worldwide. The statin therapy is one of the most effective strategies for lowering plasma low-density lipoprotein, thereby preventing atherosclerosis and reducing the incidence of heart attacks. The statin group of drugs, which include Zocor, Lipitor, and Crestor, competitively inhibit the activity of the endoplasmic reticulum (ER)-localized enzyme HMG CoA reductase. The reductase catalyzes the reduction of HMG CoA to mevalonate, a rate-determining step in the synthesis of cholesterol as well as essential nonsterol isoprenoids such as dolichol, heme, and the farnesyl and geranylgeranyl groups that are found attached to many cellular proteins. Despite the successful application in curbing cholesterol levels, statin therapy is limited because they disrupt feedback regulation of reductase causing the enzyme to accumulate to high levels. One mechanism for feedback regulation of reductase involves sterol-induced ubiquitination and subsequent degradation of the enzyme from ER membranes. Thus, a complete understanding of molecular mechanisms for reductase degradation will provide targets for novel cholesterol lowering drugs that will aid statins or be a better alternative to prevent atherosclerosis and coronary heart disease. An unresolved aspect of HMG CoA reductase degradation is how the enzyme is removed from ER membranes through a reaction called dislocation and degraded in the cytosol. To answer this, we reconstituted the cytosolic dislocation of reductase in a cell-free system. Characterization of this system revealed that dislocation of reductase from membranes required the in vitro additions of the oxysterol 25-hydroxycholesterol, the nonsterol isoprenoid geranylgeraniol, an energy source, and rat liver cytosol. We were also able to show that dislocation of reductase in the cell-free system was stimulated by two compounds, Apomine and SR-12813 that mimic sterols in stimulating reductase degradation in intact cells. This finding illustrates the utility of our cell-free system in the identification of new drugs that directly stimulate degradation of reductase. Moreover, establishment of the cell-free system positions us to identify proteins in rat liver cytosol required for reductase degradation. Modulating expression of these newly identified proteins may augment reductase dislocation and subsequent degradation, rendering them new therapeutic targets for coronary heart disease.

Background & Objectives: Sprint races are extremely popular in athletic competitions. Although those events have hitherto mainly been studied from a split-time perspective, at present, the nature of the adjustments in sprint mechanics and the extent to which stride parameters are altered are yet to be fully elucidated. This study was undertaken to test the hypothesis that single, maximal treadmill sprints (100 m, 200 m and 400 m) modify running kinetic-kinematic variables and spring-mass characteristics, with larger magnitude changes as sprint distance increases. Methods: Eleven physically active males performed 100-, 200- and 400-m running sprints on a validated instrumented sprint ergometer (ADAL3D-WR, Medical Development - HEF Tecmachine, Andrézieux-Bouthéon, France), which allowed subjects to run and produce speed ''freely'', i.e. with no predetermined belt speed imposed. Vertical and horizontal ground reaction forces were measured continuously (averaged every 50 m distance intervals) and used to determine stride parameters and spring-mass behavior. Results: Compared with the initial 50 m, running speed decreased (P<0.001) by 8±2%, 20±4% and 39±7% at the end of the 100, 200 and 400 m, respectively. All sprint distances (with the exception of stride length in the 100 m) induced significant (P<0.05) increments in contact (+7±4%, +22±8% and +36±13%) and swing times (+12±15%, +16±15% and +16±9%) and decrements in stride lengths (-1±4%, -5±5% and -41±2%) and frequencies (-6±3%, -13±7% and -22±8%) at the end of the 100, 200 and 400 m, respectively. Running kinetics all significantly decreased (P<0.001): mean vertical and total ground reaction forces for each step were reduced by about 2, 7 and 17% in the 100, 200 and 400 m, while net horizontal forces decreased by 14±17%, 41±21% and 121±35% on average. On the 100 m, the only spring-mass parameters to change (P<0.001) were an increased center of mass vertical displacement and a decreased vertical stiffness (+13±7% and -12±6%, respectively), with no change in leg stiffness. All the leg-spring variables significantly changed (P<0.05) over the 200 and 400 m: peak vertical forces: -6±3% vs. -14±9%; center of mass vertical displacement: +36±20% vs. +48±23%; leg compression: +8±8% vs. +8±9%; vertical stiffness: -30±11% vs. -63±20%; leg stiffness: -12±8% vs. -23±18%, respectively. Multiple linear regression analysis with best subset approach showed that 96% of the variation in speed was explained by stride length (ß=1.7) and frequency (ß=4.5) during 100 m. Stride length (ß=3.7) was also important predictor for speed during 200 m along with flight (ß=-24.9) and contact (ß=-25.6) times (r2=0.98), while leg stiffness showed a negative association (ß=-0.08) for predicting speed in the 400 m sprint model together with stride length (ß=-24.8) and contact time (ß=1.9) (r2=0.92). Conclusion: Along with speed, the magnitude of changes in running kinematics-kinetics and spring-mass behavior over short (100 m), intermediate (200 m) and long (400 m) treadmill sprint increased with sprint distance. Stride length was positively associated with speed in all sprint distances. Future research should evaluate the effects of various individualized training interventions on optimizing musculo-skeletal stiffness regulation, and their effects on sprinting speed development and maintenance.

Introduction The CXC-chemokine CXCL4 is released from alpha granules of activated platelets and is involved in platelet aggregation. This protein is a chemo-attractant for numerous other cell types and also functions as an inhibitor of hematopoiesis, T-cell function, tumor-induced angiogenesis and modulation of host inflammatory response. Due to its multiple functions, CXCL4 is a potential clinical anti-tumor target. We examined the effects of the CXCL4 gene on biological properties of a colorectal cancer cell line by in vitro functional tests. RNA interference (RNAi) methodology by means of gene specific stealth siRNA technology was used to control the expressional level of the CXCL4 gene. Aim of study * To determine the effect of CXCL4 knockdown by RNAi methodology on biological properties of CC531 cells in vitro. * To evaluate the role of CXCL4 gene in colorectal cancer and later on its metastasis in animal rat liver model. Materials and Methods * Rat colorectal CC531 cancer cells were inplanted into the rat liver. After selected time intervals, CC531 cancer cells were re-isolated and their expression profile was studied by mRNA micro-array analysis. In vitro cultured CC531 cells were used as control in this study. * Transient knockdown of the CXCL4 gene was achieved in CC531 cells by using Lipofectamine 2000 transfection reagent along with 100nM gene specific stealth siRNA. * After 24, 48 and 72 h of treatment with siRNA, the extent of knockdown of CXCL4 was determined at mRNA level by RT-PCR and qPCR. * Functional effects of CXCL4 knockdown on CC531 cells were investigated by proliferation, colony formation, migration and scratch assays. Results In vivo Expression of CXCL4 Implanted colorectal CC531 cancer cells exhibited 3-4 times lower expression in the rat liver for an initial period of 3 weeks and came back to normal level after week 4. Knock-down of CXCL4 Exposure to siRNA species reduced the expression of CXCL4 at mRNA level after 24-72h with maximum reduction of 70% after 24h. MTT Assay Following the post-transfection period, we found a moderate decline in cell proliferation by 20, 26 and 8% for 24, 48 and 72h, respectively, in knockdown CC531 cells. Colony Formation Assay We observed reduced ability of knockdown CC531 cells to produce small and large colonies. Migration Assay Significant reduction in directional migration was observed in the knockdown CC531 cells especially after 48 and 72hr. Scratch Assay Photomicrographs, obtained after 12 and 24 hours of scrape wounding showed a significant reduction of knockdown CC531 cells to cover the scratched area as compare to control CC531 cells. Conclusion Higher expression of CXCL4 contributes to enhanced cell proliferation, migration, colonization and wound healing abilities of colorectal CC531 cancer cells in vitro. Based on these preliminary results, we will also study the effect of CXCL4 in a liver cancer metastasis model to evaluate an anticipated correlation with colorectal cancer progression.

Introduction: According to the World Health Organization one in five road traffic fatalities are related to impaired driving involving drunk-driving (DD), drugs, illicit substances as well as prescription medications. The proportion of DD-related road fatality (or DDRF %) is the recommended indicator in setting road safety goals for DD prevention. The literature however is relatively silent about the availability of DDRF % in high-income (HIC), medium-income (MIC) and low-income (LIC) countries. Objective: The study assessed the availability of DDRF % in different countries and particularly with respect to their income and alcohol use patterns. This research is in line with Qatar National Research Strategy pillar H.E.1.8 (prevention of motor vehicle crashes and injuries) and the Grand Challenges Qatar. Methods: Availability of DDRF % was extracted from the two recent global status reports on road safety (2009) and on alcohol and health (2011) and assessed with respect to country income and alcohol use patterns. Results: Report on road safety included more DDRF% than report on alcohol and health (n=90 vs. 27). DDRF% was significantly (P&lt;0.01) more available in high-income countries (77%, n=30/39) versus middle income countries (52%, n=47/90) and low-income countries (28%, n=13/46). DDRF% distribution ranged from 5% to over 40%, however, we noted no differences between country&#39;s income status and the distribution of DDRF%. We did observe that DDRF%s were not available in the high-income countries of the Eastern Mediterranean Region. DDRF%s were available in 88% countries with high consumption (&lt;20% abstainers) One in two countries with moderate consumption did not report DDRF%. Conclusions: Data on the contribution of DD to fatality rates are generally inadequate, especially in HICs of Eastern Mediterranean region, MICs and LICs. These findings indicate the need to strengthen road safety data collection on DD, drugs and medication use while driving in above mentioned countries.

Background: Blood vessels are comprised of three distinct layers, the intima, media and outermost layer, the adventitia that, in medium to large vessels, is surrounded by a cushion of perivascular adipose tissue (PVAT). PVAT is comprised of discrete adipocytes containing a network of capillaries and nerve fibres as well as a variety of other cell types. A number of adipokines have been identified in the PVAT of human blood vessels, including leptin and adiponectin. Since PVAT is in close proximity with the adventitia there is the potential for this layer to influence vessel tone and therefore blood flow. Of particular interest is the beneficial role PVAT may play in blood vessels used as bypass grafts in patients requiring coronary artery bypass grafting (CABG). This study investigated the expression of leptin and adiponectin in the main vessels used as bypass conduits, which include the internal thoracic artery (ITA) and saphenous vein (SV). Methods and Results: SV and ITA samples were obtained at heart surgery - all with PVAT intact. &#39;Local&#39; subcutaneous fat (SV=calf/thigh; ITA = sternum) was also collected and used for: 1) ELISA, 2) histology and IHC vessels and, 3) mRNA expression. Human arterial and venous endothelial cell lines were established to ascertain endothelial leptin and adiponectin synthesis and regulation. Dense immunostaining for leptin was observed in the PVAT of both the vein and artery with no evidence of staining associated with endothelial cells lining the vessel lumen, the vasa vasorum or capillaries (as identified using CD31). Like leptin there was dense immunostaining for adiponectin in both the vein and the artery. However, discrete staining was also associated with endothelial cells lining the lumen of both the artery and the vein as well as to those of the vasa vasorum and capillaries embedded in PVAT. No leptin mRNA expression was detectable in either arterial or venous endothelial cell lines, however, adiponectin was present in both and appeared inducible by insulin. Conclusions: We show, for the first time, that the endothelium expresses adiponectin and that endothelial cell adiponectin appears to increase upon insulin stimulation. Whether these adiponectin concentrations are of functional importance in the endothelium and contribute to the vasodilatory capacity of grafted vessels are yet to be ascertained.

Background. Norepinephrine (NE) is a powerful regulator of various adipose metabolic and vascular functions, including vasoconstriction and tissue fibrosis. Recent data suggests that this may be through autocrine/paracrine effects on local resistance vessel function and morphology. Therefore, the aims of this study were to investigate in human sub-cutaneous and omental adipose tissue (SAT and OAT): 1) NE synthesis, 2) NE-mediated arteriolar vasoconstriction from non-diabetic versus diabetic obese subjects, 3) the induction of collagen genes and its deposition in the tissue. Methods and Results. AT from the abdominal subcutaneous and intra-abdominal greater omental depots was obtained during surgery (~5g each) and quickly transported in serum-free medium to the laboratory. The tissue was used for (i) histology and immunohistochemical analysis, (ii) dual wire myography for assessment of vascular reactivity, (iii) organ culture for assessment of NE release, (iv) protein extraction and NE ELISA, and (v) collagen gene type Iα1 mRNA expression. In the non-diabetic group: TH immunoreactivity, NE concentration, and maximal vasoconstriction were significantly higher in OAT compared to SAT (p&lt;0.05). But arterioles from OAT showed lower NE sensitivity compared to SAT (Log EC50: SAT versus OAT, -7.3&#177;0.6 versus -6.2&#177;0.6, p&lt;0.01). In the diabetic group: no significant depot differences were seen in NE synthesis or vasoconstriction. SAT arterioles showed significantly lower sensitivity to NE (10-8 M to 10-7.5 M, p&lt;0.05) compared to non-diabetic group. Collagen deposition and gene expression were greater in OAT than SAT of non-diabetics, while levels were elevated but comparable within both depots of diabetic subjects. Conclusions. Elevated, chronic, local NE synthesis in OAT of non-diabetic subjects may desensitize NE-induced vasoconstriction, as greater NE synthesis in both depots of diabetic patients may explain the abolishment of the depot-specific differences and sensitivity to NE in obese patients. High NE levels may also explain greater collagen gene expression and its deposition in the OAT and in diabetes.

Calreticulin (CRT) is a ubiquitously expressed protein in mammalian cells, with both calcium binding and chaperone activity. As such CRT is involved in quality control process during the folding and maturation of proteins in endoplasmic reticulum. Recent research in our lab showed that overexpression of CRT under control of Tie-2 promoter resulted in the development of metastatic lung adenocarcinoma. In order to examine genes involved in the development of lung cancer we carried out microarray analysis on RNA isolated from mouse lung tumors versus control lungs. In this screen we observed a significant increase (over 2 fold) in Metastasis Associated Lung Adenocarcinoma Transcript-1 (MALAT-1) long noncoding RNA (LncRNA) in the lungs of CRT overexpressing mouse models. MALAT-1 has been documented to be up-regulated in the human lung adenocarcinoma. This LncRNA has been shown to be associated with metastasis. Thus the aim of our study was to investigate the mechanism of regulation of MALAT-1 expression and role of CRT in this process. No data is available on the mechanism of regulation of MALAT-1 expression. We hypothesized that CRT as a regulator of intracellular calcium homeostasis regulated MALAT-1 expression thus inducing the development of metastatic lung adenocarcinoma in our mice model. Our data showed a significant correlation between intracellular calcium levels and expression and stability of MATAT-1 RNA level as determined using quantitative real time PCR. Treatment of adenocarcinoma cells with BAPTA-AM (to reduce intracellular calcium) resulted in a significant reduction in MALAT-1 level. Furthermore, treating the cells with thapsigargin (to elevate intracellular calcium) induced a 2 fold increase in MALAT-1 expression. We also demonstrated that knock down of MALA-1 expression using specific siRNA reduces the proliferation of adenocarcinoma cell derived from our CRT overexpressing mouse. Our data demonstrate for the first time involvement of calcium binding protein CRT and intracellular level of calcium on expression and stability of MALAT-1 that is involved in development of lung adenocarcinoma. This research was funded by Qatar National Research Fund (NPRP4-043-3-016).

Background Osteoporosis is a common skeletal disorder characterized by low bone mass and microarchetectual deterioration of bone tissue with increased susceptibility to fractures. Osteoporosis is considered a multifactorial polygenic disease. The prevalence of postmenopausal osteoporosis in Palestine, based on of BMD at femoral neck, total hip and spine was 24%, 14% and 29.7% respectively. Previous studies on the effect of genetic polymorphisms on bone mineral density (BMD) showed significant correlation between various haplotypes and mutations in the VDR and MTHFR and low bone mineral density [BMD] among Palestinian postmenopausal subjects. Estrogens are known to play an important role in regulating bone homeostasis. Estrogen act through binding to Estrogen Receptor α (ERα) which is a member of the nuclear receptor superfamily of ligand activated transcription factors. Vitamin K hydrochinon is an important cofactor for gamma carboxylation of osteocalcin, a major bone matrix protein . The reduction of vitamin K to vitamin K hydrochinon depends on the vitamin K epoxide reductase complex subunit 1 (VKORC1). Aim In the present study, correlation between specific XbaI and PvuII polymorphisms in the ERα gene and selected polymorphisms in the VKORC1 gene with low BMD and fractures risk were investigated in 345 postmenopausal Palestinian women including 165 osteoporotic, 93 osteopenic and 86 normal subjects using allele-specific polymerase chain reaction (PCR) and RFLP-PCR technology. Results The data showed significance association between the XX haplotype of the XbaI in ERα gene and lower BMD at the hip (P=0.012). Similarly the PP haplotype of PvuII ER α gene was significantly associated with lower hip BMD ( P=0.03). In addition, the 9041G allele [GG and GA] was significantly more frequent in patients with low BMD (P = 0.012). In our cohort, a genetic variation in the 3'-region of the VKORC1 gene (9041 AG and GG) was significantly associated with low BMD. The data also revealed the +2255 TT haplotype which is linked to lower activity of the enzyme was associated with low BMD while the presence of the C allele [CC or CT], linked to higher activity at this locus, was associated with normal BMD levels [P=0.02]. Conclusions These findings indicate that specific polymorphisms in the ERα gene and VKORC1 gene are correlated with variation in BMD levels among our subjects. These results along with previous data with the VDR and MTHFR genes provide evidence for the strong correlation between genetics and osteoporosis in our population may be significant for treatment decisions and screening of osteoporotic patients. The involvement of other genes variation like the osteoprotegrin and TNF superfamily member genes are underway. The overall data will eventialy be employed for direct correlation and evaluation between the genetic background of patients and the efficacy of selected specific drugs used to treat osteoporosis.