https://www.um.edu.mt/library/oar/handle/123456789/33607 2026-04-11T07:31:15Z https://www.um.edu.mt/library/oar/handle/123456789/42540 Title: Introduction of antimicrobial susceptibility testing to Campylobacter species and sub-species. Abstract: Antibiotic resistance, particularly with the fluoroquinolones and macrolide antibiotics, has now emerged globally with thermophilic campylobacters, including mainly Campylobacter jejuni and Campylobacter coli, giving rise to concerns about how these organisms have acquired such resistance characteristics, as well as consequences for human and animal treatment. Currently, Campylobacter is the leading enteropathogenic organism worldwide. Generally, in animals this organism causes no harm, but in humans it causes campylobacteriosis with a number of side effects including the Guillian-Barre virus. It is contracted through many sources such as private reservoirs and unpasteurised milk but mainly through ingested food. Campylobacter is a zoonotic disease. Malta, having the smallest amount of broilers, when compared to the rest of Europe, sports one of the highest percentages of Campylobacter isolation from chicken carcasses. This organism and the resistance associated with the strain are both transferred to humans. Therefore, resistance acquired through the ingestion of animal feed with antibiotics (as growth promoters), is transferred to humans through the zoonotic cycle. In order to establish the Maltese setting when compared with Europe, a series of procedures were carried out. Through the period of June 2008-June 2010, all Campylobacter strains (156 isolates) were collected from human stool samples received in the Bacteriology Laboratory at Mater Dei Hospital which is the main Maltese General Hospital and also the only teaching hospital in Malta. Each Campylobacter strain was subjected to a series of five main antibiotics and the MICs of each was established. Through this, sensitive/resistance patterns of each antibiotic was achieved and was compared with the European status. Mean Inhibitory Concentration data was achieved using the E-strip method and identification was done through conventional methods. It was established that the Maltese picture is on the same baseline as that of Europe, comparatively. Through the results of the 156 isolates analyzed, evidence of the main predominance of groups affected by this organism was established. The most age group affected was the youngest group (0-10 years). In addition to this, the male sub-group showed a slightly higher incidence when compared to the female group. Although the summer season showed a peak of infectious incidence, a confluent incidence of infection was determined throughout all months of the two year period of this study. When analyzing the sensitiveresistant patterns, Erythromycin, Gentamicin and Meropenem showed quite a high sensitivity in all strains. Even when a breakdown of each sub-species was analyzed, Erythromycin remained one of the most sensitive antibiotics tested against. Although Campylobacter coli showed much more resistance than Campylobacter jejuni, Erythromycin, Gentamicin and Meropenem remained on the sensitive side, with Ciprofloxacin and Tetracycline showing quite a pattern of resistance. Further analysis was done to compare resistance between the two gender groups (male vs females) and also between two main age-groups (0-15 years in one group and 16-above in another group). Through this comparison, the younger age group (0-15 years) had the highest amount of resistant strains when compared to the older generation. On the other hand, females showed a lower sensitivity trend when compared to the male group. These results are of great importance due to the fact that on a larger scale study, there may actually be a definite trend of resistance between age groups and gender. This data further emphasizes the great importance that Malta must be involved wholeheartedly in surveys and any data collection studies that EFSA may request. The objective of this study was to establish the sensitivity pattern of the two mam antibiotics used for first line treatment which are mainly erythromycin and ciprofloxacin. Due to the level of ciprofloxacin resistance noted in this study, erythromycin remains the drug of choice in the suspicion of any Campylobacter infection. Erythromycin is still mainly sensitive when compared to tetracycline, gentamicin and ciprofloxacin. Although, erythromycin is still the ideal drug to use, one must keep in mind that Campylobacter infections are generally self-limiting, and so over-use of this antibiotic is discouraged in order to retain this sensitivity in erythromycin. Description: M.SC.PHARMACOLOGY 2010-01-01T00:00:00Z https://www.um.edu.mt/library/oar/handle/123456789/40355 Title: The influence on the transcriptional regulation of chemokine receptor 3 (CCR3) Abstract: Chemokine receptor 3 (CCR3), the major chemokine receptor expressed on eosinophils, binds promiscuously to several ligands, mainly the eotaxin family of chemokines which are up-regulated in inflammatory response. CCR3 expression in airway epithelial cells has also been proposed to play an important role in airway inflammation by promoting epithelial wound repair and subsequent tissue remodeling. The promoter region of CCR3 gene has recently been characterized in the literature and contains promoter elements which include a TAT A box and motifs for transcription factors such as NF-KB, AP-l and GATA-I. In this study, we investigated the effects of drugs whose action include an anti-inflammatory component, and for which literature evidence for a transcriptional mechanism exists, on the transcriptional regulation of CCR3 expression. pGL3E luciferase-based reporter deletion constructs were generated for the 1.6kb CCR3 promoter region, using standard cloning approaches in DH5α E. Coli cells. Each promoter construct was transfected in a pulmonary epithelial cell line (A549) in microwell plate format and stimulated with drugs (glucocorticoid dexamethasone, endogenous glucocorticoid cortisol, and theophylline) in a dose dependent manner. This study has shown that CCR3 transcription in cytokine unstimulated A549 cells can be regulated by glucocorticoids and theophylline. A tri-phasic response (i.e. activating at low concentration, repressive at medium concentration and activating at high concentration) in CCR3 transcription response to dexamethasone was observed, indicating a complex transcriptional regulatory mechanism. Dexamethasone induced an nF-κB independent transcriptional repression of CCR3 in the unstimulated A549, possibly involving the interaction of the activated glucocorticoid receptor with AP-l in a trans-repressive molecular mechanism. Furthermore, CCR3 transcription response to the endogenous glucocorticoid cortisol was significantly different than that observed for dexamethasone. Our results have also shown that theophylline significantly represses CCR3 transcription in the absence of glucocorticoids. Description: M.SC.PHARMACOLOGY 2010-01-01T00:00:00Z https://www.um.edu.mt/library/oar/handle/123456789/39156 Title: Gender and access to medicines through the national health services Abstract: Gender and access to medicines through the National health services It is only recently that the issue of gender differences have prompted scrutiny and generated interest within the medical field. In the past few decades, gender differences in activities, goals and longevity were taken for granted, being quite predictable, however today this situation has changed, making gender differences more open to change (Verbrugge, 1985). The overall aim of the study is to identify whether there is a gender bias in terms of access to NHS medicines in Malta, as per diseases listed under the Fifth Schedule of the Social Security Act (Schedule V). This study aimed at determining whether selected diseases listed on Schedule V are more prevalent in males or in females. Specific literature searches to enable gender sensitive assessment were conducted using Medline®, Pub med®, Google scholar® and specific journals. To establish gender bias in terms of access to drugs on Schedule V, the Maltese national formulary was retrieved, together with the relevant drug use protocols. All information gathered was transferred in Microsoft Excel® for further analysis. International consensus guidelines were retrieved from NICE and SIGN. When not available, other international consensus guidelines were consulted including the BNF 58. To investigate the possibility of gender bias in access to drugs through omission of diseases as approved by Schedule V, a section of gender specific drug treatable diseases from the ICD-10 was analyzed. The results obtained clearly demonstrate higher gender prevalence in certain diseases (e.g. angina, chronic rheumatoid arthritis and systemic sclerosis are more prevalent in females; while congestive heart failure, Parkinson's disease, schizophrenia and hepatic cirrhosis are more prevalent in males) and no gender revalence in others (e.g. myocardial infarction and peptic ulcer disease). The available literature does not always indicate the gender predominance. In some instances this is not reported at all. Gender sensitive literature for Crohn's disease, diabetes insipidus and Huntington's chorea is not available, while there is no clear evidence to the gender prevalence for ulcerative colitis, chronic renal failure and psoriasis. For the intent of this study, various international consensus guidelines including SIGN, NICE and the BNF 58 were referred to and compared to the local formulary and the pertinent protocols to evaluate what is available and approved for use by protocol on Malta's national formulary with respect to the diseases listed on Schedule V. No gender bias in terms of access of medications for conditions listed on Schedule V was found. Following the analysis of gender specific drug treatable conditions for ICD-10 resulted in a list of conditions which are omitted from Schedule V, possibly creating a bias in terms of access of drugs for these conditions. The analysis leads to the conjecture that most of the medications listed on the national formulary are available and approved for use in diseases approved, as recommended by the international consensus guidelines taken as a reference. Consequently one can conclude that there seems to be no gender bias in terms of access to medications for the diseases approved by the Schedule V. The specific gender-related drug-treatable conditions of the genito-urinary system namely male disorders of the genital organs, inflammatory diseases of the prostate and male infertility, and female related disorders namely menopause, pelvic Inflammatory disease, disorders of menstruation and female infertility are not conditions which entitle patients to free medications through NHS. Mental health and musculoskeletal disorders (osteoporosis) are conditions with a female prevalence. This study deduced that these conditions are omitted from Malta's official disease list for which entitlement to free medications is approved. Looking at the percentage of employed women in relation to the positions occupied and wages earned, one reaches the conclusion that a high percentage of the Maltese female population is financially dependent. To add to this, another important consideration to be made is the fact that females tend to outlive males, and that most often they rely on males for economic resources, this means that a large percentage of older women are at risk of dependency, isolation and poverty (WHO, 2003). In light of these facts, together with the findings of this present study, it is plausible to conclude that the fact that the above mentioned gender-related drug treatable conditions which are omitted for the list of diseases, for which free medications are approved puts a large proportion of the Maltese female population at risk. Most of the medications needed to manage or treat the female related conditions mentioned earlier are already available for use through NHS; however these are not approved for the conditions investigated in this analysis. This clearly indicates a bias in terms of access for the female patient suffering from any of the mentioned condition. Description: M.SC.PHARMACOLOGY 2010-01-01T00:00:00Z https://www.um.edu.mt/library/oar/handle/123456789/38724 Title: High resolution mapping of a DNA locus genetically linked to febrile seizures Abstract: Epilepsy is regarded as a diverse family of disorders having in common an abnormally increased predisposition to seizures. Approximately 70 per cent of all patients with epilepsy lack an obvious symptomatic cause, and therefore are presumed to have a predominantly genetic basis for their condition. To date, a number of mutations in ion channel and neuroreceptor constituent genes have been identified in some syndromes with clear Mendelian inheritance. Moreover, various studies have shown that epilepsy syndromes tend to aggregate in families. The main aims of this study were to narrow down a previously identified linkage region on chromosome 20 by performing a linkage study using short tandem repeat markers, and to identify and sequence two genes found within this region which could be predisposing the family to the familial febrile seizures phenotype. This study was carried out on a Maltese three generation family having seven members affected by febrile seizures. All thirteen family members were genotyped for seven short tandem repeat markers found in the linkage region on chromosome 20 previously identified in a study conducted by Cassar in 2008. These markers were spaced at approximately 2cM from each other, using the deCODE map as a reference. Following genotyping, linkage analysis was performed and the two candidate genes chosen from the newly narrowed region were NKAIN4 and HRH3. Primer design for the exon/intron junctions of these two genes was carried out using NCBI's Primer-Blast designing tool and the respective reference sequences. Samples from an affected individual and a non-affected individual were sequenced. Subsequently the sequences obtained were first compared to each other and then to their reference sequences to determine any significant differences. Analysis of DNA from affected family members reduced the previously identified linkage region to about 5cM on chromosome 20q13.3. A maximum multipoint LOD score of 2.67 was obtained when 0.9 penetrance was used. This score is suggestive evidence for linkage. The identified locus has never been associated to familial febrile seizures syndrome before. This region spans a total of 84 genes, of which NKAIN4 and HRH3 were selected for further analysis. These two genes have been previously associated to epilepsy or seizures and are both expressed in the central nervous system. In fact, five different single nucleotide polymorphisms were found on analysis of the sequences obtained from the affected and non-affected individuals. These variations were characterised using the relevant online databases, however for various reasons none of these point mutations seemed to be immediately implicated in the formation of the febrile seizures observed in the family. Even though four of the observed single nucleotide polymorphisms in NKAIN4 were not found in the exons of the gene, this is not conclusive evidence that they do not confer a disease risk to the family. A literature review shows that heterogeneity in untranslated regions of a gene is likely to have an impact on protein expression in an individual, possibly leading to disease. This difference in protein expression may happen through various mechanisms, namely polymorphisms in micro RNA targets and disruption of splicing, leading to unstable mRNAs and defective protein structure. This project has identified a novel locus associated to the familial febrile seizures syndrome in a Maltese family and contributes to a better understanding of the condition. Discovery of gene mutations that predispose to epilepsies have led to the possibility of using gene therapy in the hope of overcoming common problems observed with the currently available drug therapy, especially in cases of pharmacoresistance. It is evident that the knowledge gained through such genetic studies can be applied to the search for improved antiepileptic drugs. Description: M.SC.PHARMACOLOGY 2010-01-01T00:00:00Z