Qatar Foundation Annual Research Conference Proceedings Volume 2016 Issue 1

Gold nanoparticle cores functionalized with highly oriented shells of oligonucleotides, referred to as spherical nucleic acids (SNAs), are novel three-dimensional oligonucleotide structures with unique properties that are distinct from their linear counterparts. As a result, SNAs exhibit novel biochemical activities that enable them to address many fundamental challenges associated with basic biological processes. They have been utilized to develop a number of powerful platforms for biomarker detection due to intrinsic unusual properties such as their amplifiable light scattering properties to achieve high assay sensitivity. In addition, these nanoconjugates show high binding affinities for complementary targets (reflected in elevated melting temperatures) and narrow subsequent melting transitions relative to oligonucleotide duplexes formed from conventional DNA probes of the same sequence. This behavior can be translated into significantly higher assay specificity and sensitivity to study the role of critical biomarkers in many forms of cancer, including prostate cancer (PCa).

PCa is the most common noncutaneous malignancy among men in the United States and the second most common cause of cancer mortality. Despite its prevalence, there are no specific accurate diagnostic or prognostic biomarkers. Indeed, although serum prostate specific antigen (PSA) concentration is used as a routine screening tool for prostate cancer, up to 11% of men with a PSA <  2.0 ng/ml may still have prostate cancer, and based on the serum level alone, it is not possible to distinguish between high and low risk prostate cancers. Due to the lack of specificity with PSA-based screening and harm associated with overtreatment and overdiagnosis, the United States Preventive Services Task Force has recommended that physicians do not routinely perform PSA-based prostate cancer screening. In an effort to separate diagnosis from treatment, active surveillance for men with low and very low risk prostate cancer, which combines PSA screening with rigorous scheduled prostate biopsies, has been implemented to decrease rates of overtreatment. However, active surveillance is a potential option only in a very select group of men with low grade and low volume PCa. From studies of men that meet strict pathologic criteria to begin active surveillance, nearly 70% can avoid treatment over five years. Yet, many urologists and patients are reluctant to monitor their cancer on active surveillance due to concerns for delaying treatment or potentially missing treatment during a window of cure. For aggressive PCa, some have concluded that it is undergraded at the time of diagnosis in up to 40% of prostate biopsies due to the limited accuracy of the technique. Thus, significant discrepancies between prostatic needle biopsy and radical prostatectomy (RP) specimens may be attributed to diagnostic pitfalls. Resolving such screening paradigms can be achieved by identifying novel molecular signatures capable of discriminating aggressive forms of PCa, which could lead to avoiding unnecessary biopsies, patient anxiety, or biopsy-related complications.

Detection of molecular signatures that are indicative of molecular changes related to cancer progression would provide a means for early diagnosis of PCa. MicroRNAs (miRNA, miR) are critical gene regulatory elements that are present in stable forms in serum and have emerged as potential non-invasive biomarkers for cancer diagnosis. However, advancement in analytical chemistry is required to detect low abundance miRNAs with high specificity. The research described here report the development of a novel scanometric-based miRNA profiling array, called the Scano-miR assay. This platform is highly sensitive and able to detect 1 femtomolar concentrations of miRNAs from serum and highly selective with the capability of identifying single nucleotide polymorphisms (i.e. SNPs). Indeed, it provides increased sensitivity for miRNA targets compared to molecular fluorophore-based detection systems, where 88% of the low abundance miRNA targets could not be detected under identical conditions. The application of the Scano-miR platform to high density array formats demonstrates its utility for high throughput and multiplexed miRNA profiling from various biological samples.

To assess the accuracy of the Scano-miR system, we studied the miRNA profiles of samples from men with PCa, and identified a novel molecular signature based on the differential expressions of circulating miRNA in serum samples specific to patients with clinically significant PCa. We analyzed the circulating miRNA profiles from patients with aggressive forms of PCa and compared them with those from healthy individuals and ones with indolent forms of the disease. From this study, a panel of five miRNAs PCa biomarkers has been identified as potentially useful for tracking both indolent and aggressive forms of the disease. Importantly, all patients with highly aggressive PCa in the study were the only cohort that exhibited elevated levels of an exclusively expressed miRNA in their serum samples. In addition, another miRNA biomarker was present in all samples from those with either aggressive or indolent forms of the disease but not detectable by qRT-PCR methods in samples from healthy volunteers. In addition, it is differentially expressed in patients with aggressive versus indolent forms of the disease. The other three identified miRNAs show different expression patterns, depending on the state of the disease. The signature is determined using a scoring method that calculates the aggregated levels of all five miRNAs by substracting the expression levels of down-regulated miRNAs from up-regulated ones and could be use to differentiate patients with aggressive forms of the diseases from those with indolent forms as well as healthy individuals with at least 94% and potentially 100% accuracy. This approach is important since it can be used to identify novel and low abundant miRNA targets for a wide variety of cancer diagnostic applications.

According to the World Health Organization (WHO), cardiovascular disease is the leading cause of deaths worldwide. The left ventricle (LV), one of the four chambers of the human heart, plays a crucial role in the performance of the entire heart. The abnormal motion of its wall muscle (myocardium) is an important indicator for multiple cardiac pathologies. Nearly, half of all heart failure cases occur due to the decline in its performance. Therefore, early detection, monitoring and accurate diagnosis of left ventricle pathologies is of critical importance. Usually, the global cardiac function parameters such as ejection fraction, and ventricular volume, etc., are used to assess the cardiac structure and functions. However, the regional abnormalities are often overlooked. The regional alterations in the heart motion are important biomarkers of several cardiac pathologies such as coronary artery disease, cardiomyopathy or hypertrophic heart disease.

The myocardium moves in a complex pattern in all three directions (radial, longitudinal, and circumferential) over the cardiac cycle. It contracts/expands (narrows/widens) and twists/untwists (clockwise/counterclockwise rotation) in short axis orientation and shortens/lengthens along the long axis direction. This complex 3D motion can be captured in a non-invasive manner using the velocity-encoded MR imaging method known as Tissue Phase Mapping (TPM). TPM offers high spatial (1–3 mm) and temporal resolution (13.8 msec) in comparison with other acquisition techniques such as tagging, tissue Doppler imaging, etc., and has proven to be a robust tool for the assessment of regional and global myocardial motion.

TPM produces spatiotemporal data that are usually visualized as a series of many static images (one for each time step). Traditionally, these static images are produced using the American Heart Association (AHA) based 17-segment model that divides the LV into 17 segments and projects them on a 2D plane perpendicular to the long axis of the heart. Also, there has been some work on the visualization of the temporal relationships but they neglect the structural or spatial information. To our knowledge, there has been no work that combines both the spatial and temporal relationships in a single representation that offers a dynamic visualization of the LV over the cardiac cycle.

In this work, we propose a novel method for the dynamic visualization of the myocardium motion over the entire heart cycle. We display both spatial and temporal relationships simultaneously for improved analysis. We propose using multiple coordinated views to show all three components (radial, longitudinal, and circumferential) of the velocity vectors in separate views (one view per component). Each component is displayed as a bulls-eye or polar plot that is color-coded with respect to its corresponding component value. The layout of the plot is such that the angular axis represents the segments of the myocardium wall, and the radial axis follows along the time dimension. The coordinated views leverage human perceptual capabilities and help in bringing out the correlations and/or disparities in the myocardial motion data. The primary objective of our work is to enhance the visual analysis to improve the understanding of the physiology and pathophysiology of the heart.

The cell metabolism and its link to oncogenic signaling pathways have got significant interest due to their importance in cancer cell analysis, anticancer drugs development, spectroscopic micro imaging of human organs and tissues and many other biomedical applications. But the lack of advanced analytical tools for the investigation of living cell metabolism is still a challenge to be faced.

Since NMR spectroscopy is a reliable analytical method which gives comprehensive and rich chemical information about the composition of unknown materials and in the same time is a nondestructive technique and high speciation performance, it is one of the major technologies for metabolic profiling, hence allows in vitro and in vivo measurements of biological cells.

Typical NMR measurements are carried out in a cylindrical 5 mm tubes with approximately 700 μL sample, but measuring small and ultra-smaller samples volumes are not possible due to the limit of detection (LOD). Moreover, ensuring the viability and the proper living condition of cancer cell using the traditional NMR detectors is critical.

We designed and fabricated a novel miniaturized planer waveguide microslot NMR detector with on-board thermal regulator integrated with a microfluidic device. A tumour spheroid in a size of a few hundred μm diameter has been studied noninvasively and in a real time investigation mode. Moreover, the NMR spectra of cellular metabolites samples fall in the 100 pmol range were obtained with this microprobe in few minutes. Additionally, the planar geometry of the detector is suitable to the size and geometry requirements such different kind of microfluidic cellular sample holders for future studies such as bio-reactors.

In our research, we focus on metabolic analysis of production\degradation rates of living human cancer cells by using the Nuclear Magnetic Resonance (NMR) of different nuclei, which give direct evidence of the present status of the cell. A dual task cellular microfluidic NMR sample holder was designed with thermal and proper gas atmosphere controlled environment in order to maintain the viability of the studied living cells at near physiological conditions for the long-term in vitro studies and the hyphenation with adaptable lab on a chip technology.

Based on the developed NMR detector and the microfluidic chip, the dynamic processes of production and degradation of 23 cellular metabolites were monitored. Remarkably high concentrations of lactate and alanine were observed, being an indicator for a shift from oxidative to glycolytic metabolism. This distinctive development has proven to be a successful analytical tool for the elucidation of cellular functions and their corresponding biochemical pathways.

In our other investigation stream, living cancer cells is treated with metal based anti-cancer drugs (such as platinum based drugs); the metabolomics respond analyses combined with the (1H, 195Pt) NMR signals and correlation times will be employed to define the mechanism of anticancer drugs action and the molecular intra and extra cellular transport mechanism, by analysing quantities of the organic and inorganic 195Pt NMR signal beside the NMR chemical shifts.

Background & Purpose

Intracellular Ca²⁺ plays an important role in the cardiac physiology and development. Augmentation in intracellular Ca²⁺ activates the hypertrophic response in cardiomyocytes, although the source of the Ca²⁺ responsible for this is still elusive. We have previously shown that calcium influx through Transient receptor potential canonical 1 (TRPC1) channel is a key mediator of the cardiac hypertrophic response and lipid raft facilitates the channels assembly. Statins have recently been shown to exert pleiotropic protective effects in cardiac hypertrophy. Here, we studied whether simvastatin interfere with the membrane lipid raft formation and affects the TRPC1 channel function to protect the cardiomyocytes against hypertrophic stimuli.

Methods

H9C2 cells were obtained from American Type Culture Collection (ATCC, Manassas, VA) and maintained in high glucose DMEM supplemented with heat-inactivated 10% fetal bovine serum (FBS) (Gibco, MD), 100 units/ml penicillin, 100 μg/ml streptomycin in a water-saturated atmosphere of 5% CO2 at 37 °C. To induce hypertrophy, cells were exposed to endothelin 1 for 48 h. Cells treated with a combination of simvastain and endothelin-1 were exposed to simvastatin 12 h prior to the addition of endothelin-1 to the treatment medium and were then incubated in the presence of both drugs for a further 48 h. protein expression was analysed by western blotting. H9C2 cells were loaded with Fura-2 to measure the TRPC1 channel function and images were taken by using EasyratioPro (PTI Tech). For transient transfection, cells were transfected with TRPC1 siRNAs or STIM1 siRNA or scrambled control siRNA using Lipofectamine 3000. To measure NFAT luciferase activity, cells were transiently transfected with pNFAT-Luc (containing firefly luciferase gene from Photinus Pyralis; Clontech, Mountain View, CA) and 0.06 μg of pRL-TK (containing Renilla luciferase gene from Renilla reniformis, used as internal control; Promega, Madison, WI). To isolate lipid raft, cells were lysed in TNE buffer and lysates were mixed with 80% sucrose (w/v), and overlaid with 6 ml of 35% sucrose followed by 4 ml of 5% sucrose (in TNE buffer). Samples were centrifuged at 34,000 rpm for 18 h at 4 °C using SW 41 Ti Swinging Bucket Rotor. Ten 1.2-ml fractions were collected from the top of the tube and used as required.

Results

Endothelin 1 treatment induced hypertrophic response in cardiomyocytes which were confirmed by measuring cell size and hypertrophic markers, ANF and BNP (60% increase compare to control). Endothelin 1 treatment significantly induced calcium overload as revealed by an increase intracellular Ca²⁺ and subsequent NFAT nuclear activation compared with control. Moreover, hypertrophic stimuli increases the expression and function of TRPC1 and activate the recruitment of TRPC1 to membrane lipid raft domain from non-raft in hypertrophic cardiomyocytes (HC). Co-immunoprecipitation of STIM1, a Ca²⁺ sensor in the sarcoplasmic reticulum (SR), revealed that the functional interaction between STIM1 and TRPC1 was increased in HC and lipid raft domain facilitates the interaction. Silencing TRPC1 or STIM1 by respective siRNAs significantly prevented the calcium overload and NFAT activation in HC. Simvastatin treatment prevent the calcium overload and attenuates the hypertrophic response in H9C2 cells. Moreover, cholesterol depletion by simvastatin markedly reduced the calcium influx via TRPC1 by affecting the lipid raft domain and decrease the interaction of TRPC1 and STIM1 in hypertrophic cardiomyocytes.

Conclusion

Our results indicate that Ca²⁺ influx through TRPC1 provides a unique source of Ca²⁺ to activate pathologic cardiac hypertrophy and blockade of TRPC1 function by affecting the lipid raft domain formation in hypertrophic cardiomyocytes may be another important therapeutic mechanism of simvastatin in preventing cardiac hypertrophy.

Mycoplasma, the smallest self-replicating microorganism, is one of the most significant problems in the cell-culture field. It belongs to the class of Mollicutes, which are characterized by the absence of a cell wall, making them resistant to commonly used antibiotics, such as streptomycin and penicillin. Because of their small size, Mycoplasmas can pass through the 220-nm pores of filters used to sterilize culture media. Owing to these criteria, contaminated cell-culture with Mycoplasma not easily detected for extended periods of time, since Mycoplasma cannot be visualized by the naked eye or even by light microscopy and do not produce overt turbid growth commonly associated with bacterial and fungal contamination. The three major sources of Mycoplasma contamination in cell-culture are 1) Cross-contamination from infected cultures, 2) contaminated culture reagents (e.g. serum and trypsin), and 3) infected laboratory personnel with M. orale or M. fermentans. Knowing the importance of cell-culture as a significant research tool for a variety of biomedical disciplines, several effective methods and kits were developed to detect Mycoplasma contamination of cell-culture and others to eliminate the contamination using antibiotics.

The frequency of Mycoplasma contamination in cell-culture has been extensively discussed in the literature. Previous reports show high incidences of Mycoplasma contamination reaching up to 35% worldwide, with extreme incidences of 65% in Argentina and 80% in Japan.

Mycoplasma contamination of cell-culture has diverse and comprehensive consequences, ranging from unsafe biological products, to erroneous experimental results. Due to the lack of amino acid biosynthesis genes, Mycoplasma competes with its' host cell nutrients and biosynthetic precursors, leading to DNA, RNA and protein synthesis alterations. Using microarray platforms, numerous gene-expression studies have shown that Mycoplasma contamination significantly up regulates and down regulates thousands of genes like oncogenes, tumor suppressors, cytokines and growth factors. Therefore, in gene-expression studies, it is critical to ensure that transcriptional changes due to Mycoplasma contamination in cell-culture are eliminated and gene-expression profile is restored to normal level after anti-Mycoplasma treatment, to avoid biased results.

For this purpose, first, we developed a survey to investigate the incidence of Mycoplasma contamination in cell-cultures and the techniques frequently used for the detection and elimination of Mycoplasma in research laboratories at Qatar. Based on the output of this survey, SKOV3 ovarian carcinoma cells, LookOut® Mycoplasma PCR Detection Kit from Sigma and BM-Cyclin from Roche were selected as most commonly used cells, detection kit and elimination kit of Mycoplasma, respectively.

Second, we used RNA-seq technology, for the first time, to investigate the transcriptional changes of Mycoplasma contamination in cell-culture and the ability of anti-Mycoplasma treatment in reversing these changes.

RNA samples extracted from uncontaminated (control), Mycoplasma-contaminated and anti-Mycoplasma treated cultured cell (Skov3) were sequenced on illumine 4000 Hi-seq. Generated data were analyzed by comparing gene expression profiles of these three conditions and summarizing all differentially expressed genes and their associated pathways. Preliminary data demonstrated that mycoplasmas alter the expression of hundreds of genes in cultured Skov3 cells. A variety of pathways are affected, and both up-regulation and down-regulation were seen. Deeper analysis specifying significantly altered pathways are in progress along with analyzing data of the post anti-Mycoplasma treatment.

The Supreme Council of Health in Qatar aims to improve its health care system by strengthening the capabilities of its health care workforce. A university in Qatar supports this national objective by offering a 2 year Bachelor of Nursing program to post-diploma nurses. The attributes gained through baccalaureate level study, particularly clinical reasoning skills and critical analysis are important for nurses to effectively participate in health care delivery, improve patient experiences and outcomes (Aiken et. al., 2014) and advance into leadership roles.

The primary purpose of this funded study was to explore the experience of post-diploma nurses returning to Bachelor level study in Qatar. A secondary purpose was to develop the research skills and capacity of undergraduate nursing students through a collaborative research approach under the mentorship of an experienced research team from the university nursing faculty and a research active, health care industry partner. Enhancement of attitudes and beliefs toward the value of research may occur as team members work toward the common goal of completing this research. Participating in a funded research project that is relevant to nursing education and health care institutions in Qatar will undoubtedly increase mentors and student researchers personal investment and commitment to the Qatari research culture and community.

This descriptive qualitative study used a Focus Group approach to explore the experience of post-diploma nurses returning to study for a degree in nursing. A volunteer sample of 19 post-diploma nurses participated in this study.

A brief demographic survey was completed and to stimulate the participants’ thoughts related to their own personal experience about returning to school, they completed a ‘Reflective Tool’ prior to focus group participation. Five focus groups were conducted co-facilitated by student and faculty research team members. The interviews were transcribed verbatim and data were analyzed using the Framework Approach (Gale, Heath, Cameron, Rashid, & Redwood, 2013).

A collaborative approach was embedded in analysis. Initially all members read and re-read the transcripts to become familiar with the data set. One interview was initially coded reaching line-by-line agreement of the whole research team. This produced a working analytical framework with tentative codes and descriptions. These were used by analysis teams (two students and a faculty mentor) to undertake initial coding of remaining transcripts; another researcher coded one transcript independently. Any additional codes generated with descriptions were added to the analytical framework. Emergent thematic categories were used to clarify and collapse codes. Subsequently all data was charted against emergent categories to generate understanding of the post- diploma nurses’ experience. Data amenable to descriptive quantitative analysis were derived from the demographic information and reflective tool.

Preliminary findings from this study provide understanding of the motivations and challenges of post-diploma nurses returning to study. For example the significance of support from the university and sponsoring health care institution is emerging as vital to academic performance and success. The contribution of this work is the insights it offers to nurse educators working with international post-diploma nurses returning to study and organizations seeking to manage part-time study of its employees. This project will contribute to the international knowledge about this topic by adding a unique Qatar context. Dissemination of research results to the international nursing community will help raise Qatar's international research profile.

References

Aiken, L.H., Sloane, D.M., Bruyneel, L., Van den Heede, K., Griffiths, P.,… Sermeus. W. (2014). Nurse staffing and education and hospital mortality in nine European countries: a retrospective observational study. The Lancet, 1362631-8, p. 1824-1830. doi: 10.1016/S0140-6736(13)62631-8.

Gale, N., Heath, G., Cameron, E, Rashid, S., and Redwood, S. (2013). Using the framework method for the analysis of qualitative data in multi-disciplinary health research. BMC Medical Research Methodology, 13:117. doi:10.1186/1471-2288-13-117

Neuroblastoma is a childhood cancer that is frequently treated with CDDP. Upon chemotherapeutic treatment, neuroblastoma patients might develop drug resistance. A large body of evidence associates the cytotoxic effect of CDDP in cancer cells with the formation of DNA adducts and thus interference with the DNA replication of cancer cells but also with the increase of the intracellular calcium concentration ([Ca²⁺]i) that leads to apoptosis. Nevertheless, it is not understood how epigenetic mechanisms, such as DNA methylation and chromatin modification might influence the effect of CDDP on cytotoxicity and [Ca²⁺]i of neuroblastoma cells.

Therefore, here we investigate in human neuroblastoma cell lines SH-SY5Y and IMR-32 the changes in cytotoxicity and calcium homeostasis induced by the treatment with epigenetic modulators: TSA and 5-AZA but also in combination with CDDP.

Treatment schemes for calcium homeostasis experiments were performed in four different groups: A) treatment with CDDP only (control); B) acute treatment with 5-AZA or TSA (without CDDP); C) pre-treatment with both 5-AZA and TSA (before CDDP application), D) pre-treatment with 5-AZA (before CDDP). For cytotoxicity test: the Trypan Blue Cytotoxicity test was performed using the automated cell counter ViCellXR (Beckman Coulter, Germany), for exposure times ranging 24–72 h. The concentrations used were for 5-AZA ranging 1–200 μM while for TSA 1–10 μM were used.

The results showed significant increase of neurblastoma cells cytotoxicity upon treatment, especially for the combinatory treatment 5-AZA and TSA. We observed a time and concentration dependent increased cytotoxicity. The most efficient treatment was the combination 5-AZA and TSA, showing additive effects. TSA showed higher efficiencity triggering cytotoxic effects in neuroblastoma cells than 5AZA. Furthermore, the combinatory treatment of neuroblastoma cells with epigenetic modulators and CDDP increased the efficiency of CDDP treatment in a synergistic manner, effect that could be confirmed in IMR32 cells. Thus, the combination 5-AZA/CDDP or TSA/CDDP or 5-AZA /TSA/CDDP was more effective in triggering cell death of neuroblastoma cells than each of the compound alone.

For the investigation of changes in [Ca²⁺]i we performed calcium imaging studies using the calcium sensitive dye Fluo-4-AM in Tyrode's buffer with calcium (1.5 mM). Fluorescent images were taken with Olympus Microscope BX51 Wi with Xenon Arc Burner and “Xcellence rt” software. CDDP (1 μM) treatment (control group) increased the [Ca²⁺]i (30.3% ± 11.7) group “A” in SH-SY5Y cells. This effect of CDDP was reversed by the pretreatment with 5-AZA and TSA group “C” (5.1% ± 5.1) (p

Viruses, Bacteria and Fungal pathogens have been responsible for destruction and degradation of plants worldwide. Most often, these pathogens live a symbiotic relationship with the plant, enhancing plant growth and at times as predators, causing a wide variety of diseases. Date palms are cultivated widely in the Middle Eastern region due to their multi-faceted uses and are valued most for their nutritious fruit – the date. The trees flourish in deep sandy loam soils and can also grow in salty and alkaline soils. An old Arab folk saying “Its feet in water and its head in file” aptly describe Date Palm trees. Therefore, the importance of Date Palm trees to Qatar and in broad to the region (Arabian Peninsula) cannot be undermined.

Date palm cultivation is hindered by several constraints; especially it's vulnerability to a wide range of bacterial, viral and fungi pathogens present in the environment. Outbreaks of diseases like Bayoud disease caused by Fusarium oxysporum was reported in Morocco and spread to neighboring countries Algeria and Tunisia, resulting in destruction of millions of trees since its outbreak a century ago. This outbreak impacted socio economic conditions of farmers as poor quality but disease resistant cultivars started dominating the landscape at the expense of commercial cultivars. In Qatar, localized incidences of disease in Date Palm trees have been reported. In 2003, Department of Agricultural Development reported a first instance of Neck bending disease on Date Palm trees growing alongside the road of Majlis AL-Taawin located in Doha, Qatar. Neck Bending was previously reported in Iraq. Therefore, continuous efforts are needed to scan for opportunistic environmental pathogens, which in future can lead to disease outbreaks. In this work, we present our findings of environmental pathogens detected with our new Date-Pathogen pipeline (DPP) using geographical sampling of Date Palm Trees covering all municipalities in the State of Qatar.

A total of 96 leaf samples were sequenced on HiSeq2500 using libraries prepared by Genotyping-by-Sequencing methodology. We identified 4 different variants of the virus Enterobacteria spp in 31 samples. They belonged to two municipalities, Al Wakrah and Al Rayyan region. We uncovered two different bacteria namely Propionibacterium acnes and Staphylococcus epidermidis in a total of 52 samples growing in the Al Rayyan Municipality region in Qatar. Eight different variants of Propionibacterium was found in all the samples except one. The above bacterial pathogens are causative agents to cause acnes in human skin and known to have no effects on plants. A recent study in 2014 has reported an example of inter kingdom bacterial host transfer involving human pathogen Propionibacterium acnes Zappae and grapevine plants. This could suggest that the strain of Propionobacterium acnes identified in our samples might be in future case of interkingdom transfer from human to date palm trees. Two fungal pathogens – Saccharomyces cerevisiae and Fusarium graminearium were also found in 42 samples. It has been shown that Saccharomyces cerevisiae produces plant hormone indole-3-acetic acid (IAA), which in sufficient quantity triggers fungal cells to be more infectious in host organism. This infection was found in trees from Al Wakrah and Al Raayan municipalities. Fusarium graminearium are known to cause fusarium head blight in wheat, barley and oat. Previously, other Fusarium species oxysporum have been found to cause Bayoud disease in date palm trees which have resulted in degradation of date palm trees in Morocco and Algeria. However, Fusarium graminearium is reported in Qatar for the first time in date palm trees growing at Doha, Al Khor, Al Daayen, Al Shamal and Umm Salal municipalities. The above results are preliminary analysis and will be explored further in the next few months.

A lot of strategies for smart approaches on surfaces were applied such as hydrogel layer, polymer brushes or self-assembly monolayers (SAM). [1] Nowadays switchable zwitterionic materials consisting of molecules with internally balanced charge between positive ammonium and negative carboxy group are promising candidates for this application. [2] They can combine antifouling properties of their zwitterion state and complexation or sticky character in their pre-zwitterionic carboxybetaine ester form. Zwitterionic forms possess antibiofouling properties due to electrostatic interaction between charged moieties, highly hydration capability and overall neutral charge in material as well as biomimetic character because zwitterions are structural similarity to biomembranes. We showed that modifications of surface by zwitterionic based self-assemble monolayer allow enhance detection limit of biosensors down to 10–15 M for analyte, [3,4] or improve electrorheological response. [5]

Carboxybetaine esters have cationic character and permit complexation with polyanionic bioabsorbents as well as character of counter ion can adjust wettability and interaction with biomolecules.

These studies will present on the utilization of pre-zwiterionic molecules: carboxybetaine based derivates formed from lipoic acid precursor in order to modify surface for construction of impedimetric lectin biosensors and for tuning wettability and interaction with DNA and other charged (bio)molecules.

Novel pre-zwitterionic carboxybetaine ester (hydrolysable and photolysable) derivates were synthetized by protocol consists of several synthetic steps and fully characterized. Subsequently, modification of a gold surface was performed by a self-assembled monolayer deposited from a solution containing prezwitterion molecules. Self-assembly monolayer, formed from derivates, was characterized by set instrumentation as atomic force microscopy, quartz crystal microbalance XPS, contact angle etc.

Hydrolysable carboxybetaine derivate was able to from complex with polycationic DNA molecules to preconcentrate and release at pH dependent manner. During course of hydrolysis carboxybetaine ester is transferred to carboxybetaine zwitterionic form to promote DNA release due to formation of carboxylate negative charge. Additionally, gradient in wettability can be observed within progress of hydrolysis and present of long perfluorinated or aliphatic types of counter ions. For example switch in wettability can be achieved only by simple and rapid couterion exchange between superhydrophilic (contact angle (CA) below 10° (to very high hydrophobilic (CA over 140°) on rough gold surface. After completed hydrolyses zwitterionic surface can be utilized as a platform for biosensor surface with nonfouling properties. Carboxylic functionality allows immobilizing sensing molecules as lectins for electrochemical impedance spectroscopy by means of EDC/NHS chemistry. This methodology provides opportunity for ultrasensitive detection up to 10–15 M of lectins which may result of a biomarker discovery on several diseases in whole media.

Moreover utilization of photolabile ester of carboxybetaine derivates allowing spatially control wettability and pattering with photomask was performed. Photolabile 2-nitrophenyl methyl ester group was introduced to pre-zwitterionic molecule and after irradiation of prepared surface with light at 365 nm was transformed from carboxybetaine ester group to zwitterionic carboxybetaine. Progress of photolysis can be observed by change of surface zeta potential, quartz crystal microbalance and contact angle measurement. This irreversible switch along with different interaction of biological species before and after photolysis will be discussed in this contribution as well.

This contribution was made possible by NPRP grant 6-381-1-078 from the Qatar National Research Fund (a member of the Qatar Foundation). The statements contained are entirely the responsibility of the authors.

References

[1] Barner, H. G.; Lutz, J.-F.; Wischerhoff, E.; Badi, N.; Laschewsky, A.; Lutz, J.-F., Smart Polymer Surfaces: Concepts and Applications in Biosciences in Bioactive Surfaces, eds. Borner, H.;Lutz, J.-F., Springer Berlin Heidelberg, 2011, 240, pp. 1–33.

[2] M. Ilcikova, J. Tkac, P. Kasak, “Switchable materials containing polyzwitterion moieties.” Polymers, accepted

[3] T. Bertok,; A. Sediva,; J. Filip,; M. Ilcikova, P. Kasak, D. Velic, E. Jane, M. Mravcová, J. Rovenský, P. Kunzo, P. Lobotka, V. Smatko, A. Vikartovska, “ Carboxybetaine interface for electrochemical glycoprofiling of antibodies isolated from human serum” Langmuir, 2015, 31, 7148–7157.

[4] T. Bertok, L. Klukova, A. Sediva, P. Kasak, V. Semak, M. Micusik, M. Omastova, L. Chovanová, M. Vlček, R. Imrich, A. Vikartovska, J. Tkáč Ultrasensitive Impedimetric Lectin Biosensors with Efficient Antifouling Properties Applied in Glycoprofiling of Human Serum Samples, Anal. Chemistry 2013, 85 (15), 7324–7332.

[5] M. Ilcikova, M. Mrlik, V. Babayan, P. Kasak, Graphene Oxide Modified by Betaine Moieties for Improvement of Electrorheological Performance, RSC Advances, 2015, 5, 57820–57827.

Introduction

Organ donation and transplant is still an evolving field in Qatar. In Qatar, a qualitative study to understand the perspective of the healthcare workers, towards barriers, promoters and system level challenges in organ donation and transplant was lacking. Hence, very limited literature is available on these issues as are experienced by healthcare workers actually involved in the various stages of this process.

Objectives

The objective of the present study was to conduct a qualitative study using phenomenological approach, with the help of focus group discussions to explore (1) Transplant system level issues; and, (2) To understand why people choose to or not to register as organ donors.

Methods

Several key stakeholders in the healthcare sector were included in the discussions. Participants were healthcare professionals (a) who are involved in organ donation and transplant activities (coordinators, surgeons, physicians), and (b) healthcare professionals involved in organ donation promotion campaigns in Qatar.

An experienced moderator from the research team was employed to conduct the discussion and the trained research assistants collected the data. The audio recordings were transcribed by professional transcribers, coded using NVivo software, analyzed in the light of Theory of Planned Behavior and researches in the similar field, and peer reviewed to derive a conclusion.

Results

The study was able to uncover several gaps in the system that are impacting the consent process and leading to under-utilization and wastage of available organs. Some key system level issues identified during the discussions were communication gap between transplant committee and some of the departments doing transplants, absence of multidisciplinary teams for organ assessment and participating through various stages of transplant process, difficulties arising because of lack of centralized centers for organ donation and transplant where all formalities could be carried out from start till the end and training deficiency reported by campaign volunteers as well as coordinators besides others. Besides this, the study was able to enlist the difficulties faced by the healthcare workers working in field of donor registrations and transplant. The study also brought out volunteers' views based on their direct interaction with public, on why people choose to or not to register during the organ donation campaigns in Qatar. Finally, the study identified some concerns in the process of organ donation and transplant where formulating new policies and protocols or amending existing ones, could affect the efficiency positively.

Conclusion

The study concludes that most challenges in organ donation and transplant in Qatar can be dealt with by focusing on creating awareness and educating people about the various issues related to organ donation through continuous campaigns and extensive media coverage of the issue. Consents, which are the core issue behind the gap between brain death cases culminating into donors, can be improved by ensuring early communication about donation decision by the donor to his/her family. Also, under-utilization and wastage could be reduced by transplant committee representation from relevant departments involved in transplants, and having multidisciplinary teams to assess the deceased donors' organs and work through the entire transplant process. This study can be referred to for further policy making in the area of organ donation and transplant in Qatar and modifying certain aspects of campaigns to make them more effective.