In the context of pharmaceutical development, it is abundantly clear that there is a need for greater understanding and control of crystalline phases. The field of crystal engineering is poised to address such issues and has matured into a paradigm for the supramolecular synthesis of new compounds with desired properties. Crystal structures are unpredictable by nature, however, the interactions that lead to crystal formation are becoming much more predictable. By means of model compound studies, the delineation of the hierarchy of hydrogen bonding between complementary functional groups or supramolecular heterosynthons can be accomplished. Competitive co-crystallization studies along with data extracted from the Cambridge Structural Database (CSD) can be utilized in understanding the reliability of supramolecular heterosynthons without the need for endless co-crystallization experiments. In effect, this ability to understand supramolecular heterosynthons can allow crystal engineers to rationally design co-crystals with a high rate of success. It has been suggested that pharmaceutical co-crystals could play a significant part in the future of API formulation since in principle they will outnumber pharmaceutical salts, polymorphs and solvates combined. The focus of this thesis is the understanding of the primary amide functional group and its hydrogen bonding capabilities; as well as the synthesis of model compounds in order to develop a blueprint for the design of pharmaceutical co-crystals using APIs that contain a primary amide functional group.

No description available.

Η στεφανιαία νόσος (ΣΝ) αποτελεί τη συχνότερη πάθηση του καρδιαγγειακού συστήματος. Κύρια αιτία της νόσου είναι η αθηροσκλήρυνση, που σήμερα θεωρείται πλέον μια χρόνια φλεγμονώδης αντίδραση του αγγειακού τοιχώματος. Η λειτουργικότητα του ενδοθηλίου του αγγειακού τοιχώματος που αποτελεί προγνωστικό δείκτη καρδιαγγειακών συμβαμάτων και μέθοδο ελέγχου της ανταπόκρισης σε φάρμακα μπορεί να προσδιοριστεί με μέτρηση της αγγειοδιαστολής που διαμεσολαβείται από τη ροή (Flow-Mediated Dilation, FMD) στη βραχιόνιο αρτηρία. Σημαντικό ρόλο στην παθοφυσιολογία της αθηροσκλήρυνσης ασκεί το σύστημα Ρενίνης-Αγγειοτενσίνης-Αλδοστερόνης (Renin-Angiotensin-Aldosterone System – RAAS) που διακρίνεται σε ενδοκρινές και ιστικό. Κεντρικά σημεία του συστήματος RAAS που αποτελούν και φαρμακευτικούς στόχους είναι η μετατροπή της Αγγειοτενσίνης Ι (ΑΤ Ι) σε Αγγειοτενσίνη ΙΙ ( ΑΤ ΙΙ ) με τη δράση του μετατρεπτικού ενζύμου της ΑT Ι (Angiotensin Converting Enzyme-ACE) και η επίδραση της ΑΤΙΙ στους υποδοχείς της. Στο ιντρόνιο 16 του γονιδίου του ενζύμου ACE(17q23) έχει βρεθεί ο πολυμορφισμός I/D που προκύπτει από την παρουσία ( Insertion– I) ή την απουσία (Deletion–D) μιας Αlu αλληλουχίας μήκους 287 bp, δημιουργώντας τρείς διακριτούς γονότυπους: II, ID και DD. Οι ομοζυγώτες DD παρουσιάζουν αύξηση κατά 50% των επιπέδων του ACE στον ορό σε σχέση με τους ομοζυγώτες II. Οι ετεροζυγώτες ID εμφανίζουν ενδιάμεσα επίπεδα. Με δεδομένο το ρόλο του συστήματος RAAS στη ΣΝ, ο ρόλος του πολυμορφισμού ACE I/D έχει καταστεί αντικείμενο μελέτης ως προς την εκδήλωση στεφανιαίας νόσου και την ανταπόκριση στεφανιαίων ασθενών στα διάφορα θεραπευτικά σχήματα αποκλεισμού του συστήματος RAAS. Στην παρούσα μελέτη προσδιορίστηκε ο πολυμορφισμός ACE I/D σε 100 φυσιολογικά άτομα και σε 100 στεφανιαίους ασθενείς. Το συμπέρασμα μετά τη στατιστική ανάλυση ήταν ότι ο γονότυπος DD δεν συνδέεται στατιστικώς σημαντικά με στεφανιαία νόσο στον ελληνικό πληθυσμό που εξετάστηκε (p>0,05). Επιπλέον προσδιορίστηκε ο πολυμορφισμός ACE I/D σε 30 στεφανιαίους ασθενείς που έλαβαν διάφορα θεραπευτικά σχήματα αποκλεισμού του συστήματος RAAS. Η ανταπόκριση των ασθενών στην αγωγή εκτιμήθηκε μέσω της μεταβολής της FMD. Ο μικρός αριθμός των ασθενών και η μεγάλη τυπική απόκλιση στις μεταβολές της FMD δεν επέτρεψε την εξαγωγή ασφαλών συμπερασμάτων για το ρόλο του πολυμορφισμού στην ανταπόκριση των ασθενών στην φαρμακευτική αγωγή. / The Renin-Angiotensin-Aldosterone System (RAAS) is an important factor for the pathogenesis of Coronary Artery Disease (CAD). The key component of RAAS is the Angiotensin Converting Enzyme (ACE). An Insertion/Deletion polymorphism (I/D) has been identified in ACE gene which accounts for half the variance of serum ACE levels (1). Subsequent reports investigated the relationship between the D allele and cardiovascular diseases, icluding CAD, with conflicting results (2). We determine ACE I/D polymorphism genotype in 100 normal individuals and 100 patients with CAD of greek origin. No association was found between D allele and CAD. Moreover, we determine ACE I/D polymorphism in 30 patients with CAD who were in treatment inhibiting RAAS. No association was found between ACE I/D polymorphism and respond.

Ο σκοπός της παρούσας διπλωματικής εργασίας ήταν να διερευνήσει την ενδεχόμενη συσχέτιση του γενετικού πολυμορφισμού -308 (G→A) του προαγωγέα (promoter) του γονιδίου του TNFα με την κλινική υποτροπή ή αγγειογραφική επαναστένωση, σε 40 Έλληνες ασθενείς μετά από αγγειοπλα-στική των στεφανιαίων αρτηριών (PTCA). Στη μελέτη περιελήφθησε και ένας πληθυσμός 30 υγιών μαρτύ-ρων. Η παρούσα μελέτη αποσκοπούσε ακριβώς στο να διερευνήσει τη συχνότητα της κλινικής υποτροπής ή αγγειογρα-φικής επαναστένωσης 6-8 μήνες μετά από PTCA, σε πληθυσ-μούς της περιοχής μας σε σχέση με το γονότυπο των ασθενών αυτών για το παραπάνω γονίδιο. Η πλειοψηφία των ασθενών 95% (38/40) ήταν ομόζυγοι για τον άγριο τύπο W του αλληλομόρφου και είχαν τον γονότυπο WW, 5% (2/40) ήταν ετεροζυγώτες WM, ενώ δεν ανευρέθησαν ομοζυγώτες MM για τον πολυμορφισμό. Στο δείγμα των 30 υγιών μαρτύρων, 90% (27/30) ήταν ομόζυγοι για τον άγριο τύπο W του αλληλομόρφου και είχαν τον γονότυπο WW, 10% (3/30) ήταν ετεροζυγώτες WM, ενώ δεν ανευρέθησαν ομοζυ-γώτες MM για τον πολυμορφισμό. Αγγειογραφική επαναστέ-νωση συνέβη σε 7 (οι 6/7 ασθενείς έχουν WW γονότυπο και ο 1/7 έχει τον WM γονότυπο) από τους 40 ασθενείς. Στην παρούσα μελέτη, δεν παρατηρήθηκε στατιστικά σημαντική συσχέτιση του -308 γενετικού πολυμορφισμού του TNFα με την αγγειογραφική επαναστένωση ή την κλινική υποτροπή μετά από αγγειοπλαστική των στεφανιαίων αρτηριών (PTCA), δείχνοντας ότι ο -308 γενετικός πολυμορφισμός του TNFα δεν είναι από μόνος του ένας βασικός παράγοντας κινδύνου, τουλάχιστον στον Ελλαδικό πληθυσμό με στεφανιαία νόσο. Βέβαια, τα αρνητικά μας ευρήματα σχετίζονται άμεσα με τη χαμηλή συχνότητα εμφάνισης του συγκεκριμένου πολυμορφισμού στους διάφορους πληθυσμούς, αλλά και με τον πολύ μικρό αριθμό ατόμων των παρατηρήσεων. / A follow-up study was conducted to investigate whether -308 genetic polymorphism of TNF-α gene was associated with the increased risk of restenosis in 40 Greek coronary artery disease patients undergoing coronary angioplasty and stent implanta-tion. For comparison of genotype frequency, a control group of 30 asymptomatic individuals was also studied. The end-point of the current study was the incidence of restenosis at 6-8 months of clinical follow-up. The majority of patients (38/40) had the WW genotype (homozygous for the wild-no polymorphic type allele) and only 2/40 patients had the WM genotype (heterozygous). Patients homozygous for the polymorphism (MM genotype) were not found. The frequency distribution was not different from that of the control subjects. Restenosis occurred in 7 of the 40 patients. In the population studied, -308 genetic polymorphism of TNF-α gene was not found to predispose patients to an increased incidence of restenosis. Nevertheless, these findings should be considered as preliminary, taking into account the small number of patients that were studied and the rarity of the -308 genetic polymorphism of TNF-α gene.

Prostate-specific antigen (PSA) testing has caused a steep increase in the incidence of prostate cancer, especially the incidence of localised low risk disease. In order to decrease the overdiagnosis accompanied by PSA testing, analysis of inherited genetic variants have been suggested as potential tools for clinical assessment of disease risk. With the aim of minimizing overtreatment and postpone side-effects of curative treatment for low risk prostate cancer, active surveillance, a treatment strategy with initial surveillance and deferred radical prostatectomy at the time of progression has evolved.  The aim of this thesis was to study the validity of PSA (paper I) and inherited genetic variants (paper II) for early diagnosis of prostate cancer, to assess the extent of PSA testing in Sweden (paper III), and to study the safety of deferred radical prostatectomy in localised low to intermediate risk prostate cancer (paper IV). The study designs were i) case-control studies nested within the Västerbotten intervention project (paper I and II), ii) observational study in the Cancer Register of Sweden (paper III), and iii) observational study in the NPCR Follow-up study (paper IV). PSA had a high validity in predicting a prostate cancer diagnosis with an area under the receiver operating characteristics (ROC) curve of 0.86 (95% CI, 0.84 to 0.88). A combined test, including PSA, the ratio of free to total PSA, and 33 single nucleotide polymorphisms (SNPs) in a genetic risk score, increased the area under curve to 0.87 (95% CI, 0.85 to 0.89). The estimated uptake of PSA testing among men aged 55 to 69 years increased from zero to 56% between 1997 and 2007 and there were large variations in the uptake of PSA testing between counties in Sweden. After a median follow-up time of eight years there was no significant difference in presence of any one or more adverse pathology features or prostate cancer specific mortality after primary compared to deferred radical prostatectomy in localised low to intermediate risk prostate cancer. Results from these studies indicate that PSA and the hitherto identified SNPs are not suitable biomarkers in single-test prostate cancer screening. It is possible to estimate the uptake of PSA testing on a population level. Initial surveillance and deferred radical prostatectomy represent a feasible treatment strategy in localised low to intermediate risk prostate cancer.

No description available.

Introduction: As a part of the Gene Regulators in Disease project (GRID), this study aims to create a novel high throughput method to discover the genetic effect on gene translation, taking advantage of the rationale that efficiently translated mRNAs associate with multiple ribosomes, while less active ones with fewer or none. / Methods: Lymphoblastoid cell lines (LCLs) from 44 HapMap European individuals were used for polyribosomal fractionation and establishing the sample bank for the future study. The fractionated mRNA samples of 10 out of the 44 individuals were run on an Illumina GoldenGate Beadarray to detect allelic imbalance (developed by the group of T.J. Hudson and T.M. Pastinen). / Results: This study established a high-quality RNA bank, including 1,100 RNA fraction samples. By the Illumina chip, translational imbalance was detected in 75 out of 1483 (5.06%) assays, and 63 out of269 (23.4%) genes. The translational effect was well replicable by the resequencing method. / Conclusion: This study found that genetic effect on gene translation is a common mechanism of expression regulation. Our best hit found in the integrin beta 1 binding protein 1 gene (ITGB1BP1 ) highlights the role of mRNA 3'UTR secondary structure in gene translation. / Keywords: Gene translation, High throughput genotyping, Human genetics, Polyribosome, RNA, Single nucleotide polymorphism

Darbo tikslas - ištirti STAT 5 geno įvairovę Lietuvoje auginamų galvijų tarpe. LVA Gyvūnų genetikos laboratorijoje įdiegta galvijų STAT 5 geno, ištyrimo metodika. Pirmą kartą ištirtas STAT 5 geno polimorfizmas Lietuvoje auginamų galvijų tarpe Darbo uždaviniai: 1. Surinkti ir išanalizuoti mokslinę literatūrą apie mėsinių galvijų STAT 5 geną ir ištirti jo įtaką produktyviosioms savybėms, ieškoti straipsnių internetinėse svetainėse nagrinėjama tema; 2. Įdiegti galvijų STAT 5 geno tyrimo metodiką LVA K.Janušausko Gyvūnų genetikos laboratorijoje; 3. Ištirti STAT 5 geno įvairovę Lietuvoje auginamų galvijų tarpe, naudojant sumodeliuotus pradmenis pagal geno seką; 4. Nustatyti STAT 5 geno aleli�� ir genotipų dažnius skirtingose galvijų veislėse. / Object and tasks of work. Analyse and summarize literature about STAT 5 gene in cattle. Introduce bovine STAT 5 gene research methodology at K. Janušauskas Laboratory of Animal Genetics, LVA..Investigate STAT 5 gene polymorphism and distribution of different alleles and genotypes in cattle breeds bred in Lithuania. Research methodology. DNA extraction from hair roots; PGR to amplify STAT 5 gene; RFLP method-STAT 5 – enzyme AvaI; Electrophoresis in agarose gel; Staining with etidium bromide; Genotyping; Statistical analysis of data. Results and conclusions. Two alleles C and T were found in tested cattle group identified by PGR-RFLP method. There was found a bit lower heterozigosity than expected in STAT 5 gene locus. The frequency of C allele was 0,89, T allele – 0,11, CC genotipe – 0,79, Ct genotipe -0,21. TT genotipe was not found. In all tested cattle breeds prevailing was found C allele and CC genotipe varying from 50 percent in Sharole breed to 100 percent in simental breed. CT heterozygote genotipe in pargest frequency 0,5 found in Sharole breed and lowest frequency 0,03 in Lithuanian black and white.

The heat capacity and the enthalpy of crystallization of the crystalline phases at the end of cooling must be known in order to determine the excess energy of mixing two pure triglycerides, trilaurin and trimyristin, cooled at different cooling rates. The present investigation was carried out using Differential Scanning Calorimetry, DSC, Modulated Differential Scanning Calorimetry, MDSC®, and Thermal Relaxation (in a Physical Properties Measurement System, PPMS). It was found that enthalpy of crystallization values can be measured to within ? 2% (SE) with DSC Q100 TA Instruments. To achieve this, an experimental procedure and a data analysis method are proposed. It was not possible in this study to obtain accurate and reproducible heat capacity values using a DSC Q100 instrument. The values were shown to be significantly by the position of the sample pan in the measuring sensor. PPMS Cp values were within the literature values.

The cost of feed remains an important factor affecting the profitability of beef production, and the difficulty of recording feed intake is a major limitation in an industry-wide selection program. Novel genomics approaches offer opportunities to select for efficient cattle. Therefore, the main objective of this work was to identify genetic markers responsible for genetic variation in feed efficiency traits as well as to understand the molecular basis of feed efficiency traits. The candidate gene approach revealed new single nucleotide polymorphisms (SNPs) in the Cholecystokinin B receptor (CCKBR) and pancreatic anionic trypsinogen (TRYP8) genes that showed strong evidence of association with feed efficiency traits. An in silico approach was proposed as a cost-effective method for SNP discovery. SNPs within genes Pyruvate carboxylase, ATPaseH+, UBQEI, UCP2, and PTI showed evidence of association with carcass traits without negatively affecting feed efficiency traits. The polymorphisms within genes CCKBR and TRYP8 were associated with pancreas mass and pancreatic exocrine secretion. A fine-mapping study on 1,879 SNPs revealed 807 SNPs with significant associations corresponding to 1,012 genes. These 807 SNPs represented a genomic heritability of 0.32 and 89% of the genetic variance of residual feed intake (RFI). Genomic breeding values estimated from the SNP set (807) were highly correlated (0.96) to the breeding values estimated from a mixed animal model. The 10 most influential SNPs were located in chromosomes 16, 17, 9, 11, 12, 20, 15, and 19. Enrichment analysis for the identified genes (1,012) suggested 110 biological processes and 141 pathways contributed to variation in RFI. The 339 newly identified SNPs corresponding to 180 genes identified by fine-mapping were tested for association with feed efficiency, growth, and carcass traits. Strong evidence of associations for RFI was located on chromosomes 8, 15, 16, 18, 19, 21, and 28. Combing validated SNPs from fine-mapping and the candidate gene approach may help develop a DNA test panel for commercial use and increase our understanding of the biological basis of feed efficiency in beef cattle. / The Ministry of Higher Education of Egypt