Passive loading of a specimen is a relatively cheap method to use in fracture mechanical testing compared to an actively loaded specimen. For stress corrosion cracking testing it is easier to use a passively loaded specimen since the specimen easily can be placed in a specific corrosive environment. The passive method lacks information about the crack growth over time and the load can not be regulated during the test to ensure crack growth. This thesis work was mainly about finding a specimen with a region of constant KI to ensure crack growth without the need of controlling the load and to find a way to estimate the crack growth over time. The work is based on Linear Elastic Fracture Mechanics and the Finite Element Method. The thesis work resulted in a specimen with constant KI in the region 23/50 ≤ a/W ≤ 33/50 of crack growth and an equation was found to describe the relation between the crack propagation and the strain measured on the specimens back face. / Passiv belastning av en provstav är en relativt billig metod att använda för brottmekanisk provning jämfört med en aktivt belastad provstav. En passivt belastad provstav är enklare att använda vid provning av spänningskorrosion då provstaven enkelt kan placeras i en specifik korrosiv miljö. Den passiva metoden är bristande i information om spricktillväxen över tid och belastningen på provstaven kan inte justeras för att upprätthålla fortsatt spricktillväxt under provningens gång. Detta examensarbete syftade till att ta fram en provstav med ett spricktillväxtomrade med konstant KI för fortsatt spricktillväxt vid konstant belastning samt att hitta en metod för att uppskatta spricktillväxten över tid. Arbetet är baserat på linjärelastisk brottmekanik och finita elementmetoden. Arbetet resulterade i en provstav med konstant KI i spricktillväxtområdet 23/50 ≤ a/W ≤ 33/50 och en relation mellan spricktillväxt och töjningen som mäts på provstavens baksida.

Proline-rich antimicrobial peptides (PrAMPs) are promising candidates to treat bacterial infections. The designer peptide ARV-1502 exhibits strong antimicrobial effects against Enterobacteriaceae both in vitro and in vivo. Since the inhibitory effects of ARV-1502 reported for the 70 kDa heat-shock protein DnaK do not fully explain the antimicrobial activity of its 176 substituted analogs, we further studied their effect on the bacterial 70S ribosome of Escherichia coli, a known target of PrAMPs. ARV-1502 analogues, substituted in positions 3, 4, and 8 to 12 (underlined) of the binding motif D3KPRPYLPRP12 with aspartic acid, lysine, serine, phenylalanine or leucine, were tested in a competitive fluorescence polarization (FP) binding screening assay using 5(6)-carboxyfluoresceinlabeled (Cf-) ARV-1502 and the 70S ribosome isolated from E. coli BW25113. While their effect on ribosomal protein expression was studied for green fluorescent protein (GFP) in a cell-free expression system (in vitro translation), the importance of known PrAMP transporters SbmA and MdtM was investigated using E. coli BW25113 and the corresponding knockout mutants. The dissociation constant (Kd) of 201 16 nmol/L obtained for Cf-ARV-1502 suggests strong binding to the E. coli 70S ribosome. An inhibitory binding assay indicated that the binding site overlaps with those of other PrAMPs including Onc112 and pyrrhocoricin as well as the non-peptidic antibiotics erythromycin and chloramphenicol. All these drugs and drug candidates bind to the exit-tunnel of the 70S ribosome. Substitutions of the C-terminal fragment of the binding motif YLPRP reduced binding. At the same time, inhibition of GFP expression increased with net peptide charge. Interestingly, the MIC values of wild-type and DsbmA and DmdtM knockout mutants indicated that substitutions in the ribosomal binding motif altered also the bacterial uptake, which was generally improved by incorporation of hydrophobic residues. In conclusion, most substituted ARV-1502 analogs bound weaker to the 70S ribosome than ARV-1502 underlining the importance of the YLPRP binding motif. The weaker ribosomal binding correlated well with decreased antimicrobial activity in vitro. Substituted ARV-1502 analogs with a higher level of hydrophobicity or positive net charge improved the ribosome binding, inhibition of translation, and bacterial uptake.