[Iatrogenic femoral AV-fistula].

The common femoral artery is a widely used for access in endovascular interventions. Various complications, such as hematoma, pseudoaneurysm and AV-fistula (AVF), can arise from arterial punctures with estimated prevalence between 1-10%. AVF is a rare complication with prevalence p<1%. AVF can cause a hemodynamic change in the form of a arteriovenous shunt (AV-shunt). AV-shunts in the groin are usually small and asymptomatic but tend to be symptomatic with larger and persistent AVFs which can present with leg claudication or high outpute heart failure.

[Spontaneous retroperitoneal hemorrhage].

A 50-year old male presented to our emergency department with sudden abdominal pain. Upon arrival he was diaphoretic, pale and tachycardic. A CT showed retroperitoneal hemorrhage with suspected tumor at the left adrenal gland. He was quickly stabilized with intravenous fluids and blood transfusion. Rebleed occurs roughly a week after discharge and a new CT showed a visceral pseudoaneurysm from the left middle adrenal artery. The pseudoaneurysm was embolized and the patient discharged in good condition. Follow-up MRI depicted reabsorption of the hematoma and no adrenal tumor. Thus, the etiology of the previous retroperitonal hemorrhage is considered spontaneous.

Changes of center of rotation and femoral offset in total hip arthroplasty.

BACKGROUND: It is a challenge to reconstruct the center of rotation (COR) and femoroacetabular offset anatomically in total hip arthroplasty (THA). We addressed the controversy how we manage to preserve the COR and femoroacetabular offset with an uncemented total hip prosthesis implanted with free hand technique. METHODS: We analyzed a prospective series of 73 patients who underwent primary THA. The series was composed of 40 females and 33 males, mean age 64 years (range, 35-90 years). The reasons for THA were primary osteoarthritis (n=63) and developmental dysplasia of the hip (n=10). Pre- and postoperative X-rays were done in a standardized format for anterior-posterior (AP) radiographs of the pelvis, and digitalized measurements were done using SectraTM. We compared preoperative measurements with the final outcome to determine changes in COR and femoral offset. RESULTS: We found that 40 patients had their COR and 34 patients had their femoral offset preserved within preoperative 5.0 mm limits. Twenty-three patients had both their values of COR and femoral offset preserved within 5.0 mm limits. While a significant correlation was found between changes of femoral and global offset (r=0.786, P<0.001), there were no correlation between changes of acetabular and femoral offset (r=-0.027, P=0.822). CONCLUSIONS: Using an uncemented THA and free hand technique, there is a fair reproducibility of anatomy. The variations were mostly minor, but our results indicate a potential for better restoring the location of COR and femoral offset when planning and implanting an uncemented THA.

Characterization of cold-adapted Atlantic cod (Gadus morhua) trypsin I--kinetic parameters, autolysis and thermal stability.

Atlantic cod trypsin I is a highly active cold-adapted protease. This study aimed at further characterization of this enzyme with respect to kinetic parameters, sites of autolysis and stability. For that purpose, trypsin I was purified by anion exchange chromatography. Its purity and identity was verified by SDS-PAGE analysis and mass spectrometry. Concomitantly, another cod trypsin isozyme, trypsin X, previously only described from its cDNA sequence was detected in a separate peak from the ion exchange chromatogram. There was a stepwise increase in the catalytic efficiency (k(cat)/K(m)) of cod trypsin I obtained with substrates containing one to three amino acid residues. As expected, the activity of trypsin I was maintained for longer periods of time at 15 degrees C than at higher temperatures. The residues of the trypsin I molecule most sensitive to autolysis were identified using Edman degradation. Eleven autolytic cleavage sites were detected within the trypsin I molecule. Unfolding experiments demonstrated that autolysis is a contributing factor in the stability of trypsin I. In addition, the data shows that cod trypsin I is less stable towards thermal unfolding than its mesophilic bovine analogue.

Recombinant cold-adapted trypsin I from Atlantic cod-expression, purification, and identification.

Atlantic cod trypsin I is a cold-adapted proteolytic enzyme exhibiting approximately 20 times higher catalytic efficiency (kcat/KM) than its mesophilic bovine counterpart for the simple amide substrate BAPNA. In general, cold-adapted proteolytic enzymes are sensitive to autolytic degradation, thermal inactivation as well as molecular aggregation, even at temperatures as low as 18-25 degrees C which may explain the problems observed with their expression, activation, and purification. Prior to the data presented here, there have been no reports in the literature on the expression of psychrophilic or cold-adapted proteolytic enzymes from fish. Nevertheless, numerous cold-adapted proteolytic microbial enzymes have been successfully expressed in bacteria and yeast. This report describes successful expression, activation, and purification of the recombinant cod trypsin I in the His-Patch ThioFusion Escherichia coli expression system. The E. coli pThioHis expression vector used in the study enabled the formation of a fusion protein between a highly soluble fraction of HP-thioredoxin contained in the vector and the N-terminal end of the precursor form of cod trypsin I. The HP-thioredoxin part of the fusion protein binds to a metal-chelating ProBond column, which facilitated its purification. The cod trypsin I part of the purified fusion protein was released by proteolytic cleavage, resulting in concomitant activation of the recombinant enzyme. The recombinant cod trypsin I was purified to homogeneity on a trypsin-specific benzamidine affinity column. The identity of the recombinant enzyme was demonstrated by electrophoresis and chromatography.