Drug design of novel molecules using a bioisosteric and de novo techniques - a comparison

Rational drug design is an area of science that evolves continuously in order to answer contemporary demands for a decrease in novel drug discovery turnover time. Multiple drug design modalities exist which may be exploited in response to the parameters of specific drug design projects. Bioisosteric modification of existing molecules and de novo design are two such approaches, both of which were employed in parallel in this study which aimed to compare their scope and efficiency using Tricyclic Antidepressants (TCAs) and Selective Serotonin Reuptake Inhibiting (SSRIs) molecules as case studies. Results indicated that bioisosterically modified structures did not have a higher affinity for their cognate receptor when compared to the template structure while the de novo design yielded molecules that were markedly different to the template from a structural perspective, and which also bound to the cognate receptor with an affinity superior to that of the template. This study showed therefore that bioisosteric modification is of utility when minor structural variations are considered sufficiently relative to a template molecule, and could consequently be of utility in the acquisition of new patents, in the reduction of toxicity, or in the attainment of improved biological profiles. It indicated furthermore, the role of the de novo approach in the successful exploration of novel pharmacophoric space and in the generation of molecular structures with an affinity significantly greater than that of lead molecules for a target receptor.peer-reviewe

A study of Gene, Protein and miRNA alterations in women with Endometriosis.

PhDThe aim of this thesis was to improve understanding of the underlying genetic and proteomic alterations potentially contributing to endometriosis. Assessment of the genetic mechanisms and pathways controlling angiogenesis, apoptosis and inflammation allowed identification of potential aberrations contributing to disease. Tissue miRNA expression experiments show that, the ebv-mir-BART2-5p is detected in endometriosis. Endometriosis cells contained higher levels of ebv-mir-BART2-5p compared to eutopic endometrium and this finding was confirmed by quantitative PCR. In situ hybridisation for EBV on tissue microarrays did not confirm the presence of active EBV within the endometriotic epithelial cells (Figure 4-24) but 5 of the 42 endometriotic samples on TMA-A gave a positive reading for EBV presence in some of the lymphocytes. PCR on the peripheral blood monocytes confirms overall higher levels of EBV DNA in the monocytes of people with endometriosis compared to controls (see Table 9-39 in the Appendix). There were no detected EBV levels in the surgically confirmed control patient samples. The presence of ebv-mir-BART2-5p is a permissive event for the development of endometriosis potentially acting as an initiator for engraftment of endometrial cells to the peritoneum causing the development of endometriosis. It also aids in disease development by suppressing T-cell function and encouraging adhesions and angiogenesis in affected tissues. Alterations in cellular genotype also enable the ectopic endometrial cells to evade NK cells and Lymphocytes, promoting proliferation and metastasis. Effects of genes on various stages of the cell cycle checkpoints and pathways have been explored as contributors to disease development. Downstream proteins from EBV upregulation are also confirmed to be affected. Tissue microarray studies demonstrate the upregulation of Cyclin D1 (Figure 4-22) and downregulation of E-Cadherin, Maspin and BCLAF-1) (Figure 4-11, Figure 4-13 and Figure 4-15). Galectin 3 (a prominent anti-apoptotic and angiogenic member of the lectin family) proteins were also upregulated in endometriosis. Galectin-3 had already been shown to be a good therapeutic target in humans and may provide alternatives to current surgical or hormonally repressive therapy. A set of in vitro experiments have been performed and show effective disease repression with Galectin therapeutics (Galectin 3 inhibitor GCS-100), potentially opening a window for the development of novel therapeutics. Serum was analysed for miRNA and antibody protein expression profiles. Pathways linked to identified biomarkers have been explored aiding in the understanding of disease development at a molecular level. Identified miRNAs are seen to interact with a number of important pathways listed below. There could potentially be effects on cellular mitosis and meiosis, cellular structure, intra and intercellular signalling, vesicular transport including exo and endocytosis and cellular apoptosis. All of these changes can result in endometriotic cells that replicate at accelerated rates, adhere to ectopic sites, enable angiogenesis and growth, evade normal apoptotic mechanisms and evade immune responses. Certain identified pathways act as tumour suppressors that could potentially explain the non-malignant properties in the majority of cases of endometriosis. These include the p38 MAPK1 pathway and the p53 tumour suppressor genes2. Other pathways involved are known to be associated with tumorigenesis and have pro-oncogenic properties, these include the RAS, WNT, TRK receptors and MAPKKKK pathways. It is the fine balance between tumour suppressors and oncogenes that probably control the transition between endometriosis and endometrial carcinomas. Identification of protein and miRNA expression profiles predicting the presence of endometriosis allowed us to work towards developing a panel of non-invasive biomarkers (Patent P063434GB) for earlier identification and treatment of disease and aids in unlocking new methods of molecular targeting as treatment

Erosive tooth wear in children and adolescents

To determine the local prevalence of erosive tooth wear in the child population and to identify the degree to which local demographic and socioeconomic factors influence prevalence, a multi-stage cluster sample of three, five, eight, twelve and fifteenyear old Maltese school children were identified. The children were clinically examined under standardised conditions and provided a questionnaire to be filled directly (twelve and fifteen-year-olds) or by the parents/legal guardians (three, five and eight-year-olds). A total of 2508 children were examined. Of these, 232 three-year-old, 338 five-year-old children, 337 eight year-old children, 642 twelve-year-old children and 560 fifteen-year-old children returned a questionnaire and were analysed. The prevalence of erosive tooth wear was > 70% in all age cohorts. Erosion experience also increased in both extent and severity with age in each dentition. Significant higher incidences were observed in eight-year old males, eight-year old overweight children, eight and fifteen-year-olds attending public schools, locality (Gozo > Malta), history of vomiting in fifteen-year olds, and children from lower socioeconomic parental status in five, eight and fifteen-year-olds. The prevalence of erosive tooth wear is high in school aged Maltese children. This easily preventable tooth condition deserves targeted public health programmes to improve the oral health of future generations.peer-reviewe

PRESENCE OF ANISAKIS PEGREFFII IN FARMED SEA BASS ( DICENTRARCHUS LABRAX L.) COMMERCIALIZED IN SOUTHERN ITALY: A FIRST REPORT

We examined 151 European sea bass (Dicentrarchus labrax L.) samples from farms and fish markets of Sicily (Southern Italy) for Anisakidae larvae detection. All the samples were examined by visual inspection and modified chloro-peptic digestion. Two nematode larvae were found in the viscera of only one European sea bass sample from a farm located in Greece (FAO 37.3), giving a total prevalence of infestation of 0.7%. No other parasites were found after chloro-peptic digestion of the samples. The larvae were morphologically ascribed, at genus level, to morphotypes I and molecularly identified as Anisakis pegreffii. To the best if our knowledge, this is the first report on the presence of anisakid parasites in farmed European sea bass of Mediterranean Sea. Our findings suggest that the risk of exposure to Anisakidae nematodes in farmed European sea bass remains very low. However, further data on Mediterranean farms are needed to have a detailed risk analysis

Social and Emotional Education - Building inclusive schools and ownership of values - Programme of activities

Young people need to have a balanced set of cognitive, social and emotional competences in order to help them navigate successfully through the developmental tasks, situational challenges, and transitions they are set to face in their pathway to adulthood

The Relationship of Surface Characteristics and Antimicrobial Performance of Pulp Capping Materials

Pulp capping materials need to be able to protect the pulp but also bond to the overlying restorative materials. Light-curable pulp capping materials bond better to restorative materials and are easier to place than most water-based cements. The aim of this study was to characterize new light-curable tricalcium silicate-based pulp capping materials and compare their surface and antimicrobial properties with clinically available Theracal (Bisco, Schaumburg, IL) and Biodentine (Septodont, Saint-Maur-des-Fossés, France). The surface characteristics of 3 light-curable pulp capping materials based on a resin and filled with tricalcium silicate and tantalum oxide radiopacifier and Theracal and Biodentine were assessed by scanning electron microscopy, X-ray diffraction, and contact angle measurement. The radiopacity was measured following ISO 6876 standards. The antimicrobial activity was determined by the direct contact test and the antibiofilm activity by the adenosine triphosphate assay and the confocal laser scanning Live/Dead assay (Invitrogen, Eugene, OR) using a polymicrobial culture. The surface characteristics of the materials varied with the unfilled resin and Biodentine exhibiting a hydrophobic surface. Biodentine showed significantly higher antimicrobial properties in the direct contact test, but this property was absent in the antibiofilm activity tests. The resins filled with tricalcium silicate and Theracal showed higher antimicrobial activity than Biodentine in the adenosine triphosphate and live/dead assays. The surface characteristics of a material affect its antimicrobial properties. The experimental resin-modified materials exhibited comparable antimicrobial properties with other light-curable pulp capping agents. Further long-term studies on the materials' antimicrobial activity are required to assess whether they can result in better clinical outcomes

Geographic distance, water circulation and environmental conditions shape the biodiversity of Mediterranean rocky coasts

11 páginas, 2 tablas, 3 figuras.Ecological connectivity is important for effective marine planning and biodiversity conservation. Our aim was to identify factors important in influencing variation in benthic community structure on shallow rocky reefs in 2 regions of the Mediterranean Sea with contrasting oceanographic regimes. We assessed beta (β) diversity at 146 sites in the littoral and shallow sublittoral from the Adriatic/Ionian Seas (eastern region) and Ligurian/Tyrrhenian Seas (western region) using a null modelling approach to account for variation in species richness. The distance decay relationship between species turnover within each region and geographic distance by sea was determined using generalised linear models. Mantel tests were used to examine correlations between β diversity and connectivity by ocean currents, estimated from Lagrangian dispersal simulations. Variation in β diversity between sites was partitioned according to environmental and spatial components using a distance-based redundancy approach. Species turnover along a gradient of geographic distance was greater by a factor of 3 to 5 in the western region than the eastern region, suggesting lower connectivity between sites. β diversity was correlated with connectivity by ocean currents at both depths in the eastern region but not in the western region. The influOPEN ACCESS ence of spatial and environmental predictors of β diversity varied considerably between regions, but was similar between depths. Our results highlight the interaction of oceanographic, spatial and environmental processes influencing benthic marine β diversity. Persistent currents in the eastern region may be responsible for lower observed β diversity compared to the western region, where patterns of water circulation are more variable.This work was supported by the European Community’s 7th Framework Programme (FP7/ 2007−2013) under Grant Agreement No. 287844 for the project ‘Towards COast to COast NETworks of marine protected areas (from the shore to the high and deep sea), coupled with sea-based wind energy potential (CoCoNet)’.Peer reviewe

Hepatitis E Virus Occurrence in Pigs Slaughtered in Italy

In Europe, foodborne transmission has been clearly associated to sporadic cases and small clusters of hepatitis E in humans linked to the consumption of contaminated pig liver sausages, raw venison, or undercooked wild boar meat. In Europe, zoonotic HEV-genotype 3 strains are widespread in pig farms but little information is available on the prevalence of HEV positive pigs at slaughterhouse. In the present study, the prevalence of HEV-RNA positive pigs was assessed on 585 animals from 4 abattoirs located across Italy. Twenty-one pigs (3.6%) tested positive for HEV in either feces or liver by real-time RT-PCR. In these 21 pigs, eight diaphragm muscles resulted positive for HEV-RNA. Among animals collected in one abattoir, 4 out of 91 plasma tested positive for HEV-RNA. ELISA tests for the detection of total antibodies against HEV showed a high seroprevalence (76.8%), confirming the frequent exposure of pigs to the virus. The phylogenetic analyses conducted on sequences of both ORF1 and ORF2 fragments, shows the circulation of HEV-3c and of a novel unclassified subtype. This study provides information on HEV occurrence in pigs at the slaughterhouse, confirming that muscles are rarely contaminated by HEV-RNA compared to liver, which is the most frequently positive for HEV

New Mediterranean biodiversity records (October 2015)

The Collective Article “New Mediterranean Biodiversity Records” of the Mediterranean Marine Science journal offers the means to publish biodiversity records in the Mediterranean Sea. The current article has adopted a country-based classification and the countries are listed according to their geographic position, from west to east. New biodiversity data are reported for 7 different countries, although one species reported from Malta is new for the entire Mediterranean basin, and is presumably also present in Israel and Lebanon (see below, under Malta). Italy: the rare native fish Gobius kolombatovici is first reported from the Ionian Sea, whilst the alien jellyfish Rhopilema nomadica and the alien fish Oplegnathus fasciatus are first reported from the entire country. The presence of O. fasciatus from Trieste is concomitantly the first for the entire Adriatic Sea. Finally, the alien bivalve Arcuatula senhousia is reported for the first time from Campania (Tyrrhenian Sea). Tunisia: a bloom of the alien crab ortunus segnis is first reported from the Gulf of Gabes, where it was considered as casual. Malta: the alien flatworm Maritigrella fuscopunctata is recorded in the Mediterranean Sea for the first time, on the basis of 25 specimens. At the same time, web searches include possible unpublished records from Israel and Lebanon. The alien crab P. segnis, already mentioned above, is first formally reported from Malta based on specimens collected in 1972. Concomitantly, the presence of Callinectes sapidus in Maltese waters is excluded since based on misidentifications. Greece: the Atlantic northern brown shrimp Penaeus atzecus, previously known from the Ionian Sea from sporadic records only, is now well established in Greek and international Ionian waters. The alien sea urchin Diadema setosum is reported for the second time from Greece, and its first record from the country is backdated to 2010 in Rhodes Island. The alien lionfish Pterois miles is first reported from Greece and concomitantly from the entire Aegean Sea. Turkey: the alien rhodophyte Antithamnion hubbsii is first reported from Turkey and the entire eastern Mediterranean. New distribution data are also provided for the native fishes Alectis alexandrina and Heptranchias perlo. In particular, the former record consists of a juvenile measuring 21.38 mm total length, whilst the latter by a mature male. Cyprus: the rare native cephalopod Macrotritopus defilippi, and the alien crab Atergatis roseus, sea slug Plocamopherus ocellatus and fish Cheilodipterus novemstriatus are first recorded from the entire country. Lebanon: the alien crabs Actaea savignii and Matuta victor, as well as the alien fish Synanceia verrucosa, are first recorded from the entire country. In addition, the first Mediterranean record of A. savignii is backdated to 2006, whilst the high number of M. victor specimens observed in Lebanon suggest its establishment in the Basin. The Atlantic fishes Paranthias furcifer and Seriola fasciata, and the circumtropical Rachycentron canadum, are also first reported from the country. The P. furcifer record backdates its presence in the Mediterranean to 2007, whilst S. fasciata records backdate its presence in the eastern Mediterranean to 2005. Finally, two of these latter species have been recently ascribed to alien species, but all three species may fit the cryptogenic category, if not a new one, better.peer-reviewe

Service-learning in dental education. The experience in Malta

Service learning has gained relevance in dental education in the past decades. Traditionally it is integrated in community dentistry as a key component of the dental curriculum. However emerging trends in our communities’ demand further exploration and development of this notion. Community dentistry, through service learning, provides the framework for the student to understand the concept of social responsibility. Furthermore, it enhances the educational experience in the different aspects of dental education, including improving their clinical skills, self-confidence and empathy. At the same time, it proves to benefit the communities at large by providing access to oral health care. In this paper, we present our experiences in service learning and growing as a community-engaged faculty by providing healthcare to vulnerable populations, creating programs and opening several clinical facilities, and finally earning the respect of the community. Service learning has become one of our greatest strengths as a Faculty.peer-reviewe