Anders Nygren

In situ genotyping individual DNA molecules by target-primed rolling-circle amplification of padlock probes.

Methods are needed to study single molecules to reveal variability, interactions and mechanisms that may go undetected at the level of populations of molecules. We describe here an integrated series of reaction steps that allow individual nucleic acid molecules to be detected with excellent specificity. Oligonucleotide probes are circularized after hybridization to target sequences that have been prepared so that localized amplification reactions can be initiated from the target molecules. The process results in strong, discrete detection signals anchored to the target molecules. We use the method to observe the distribution, within and among human cells, of individual normal and mutant mitochondrial genomes that differ at a single nucleotide position.

MR imaging of arrhythmogenic right ventricular cardiomyopathy: Morphologic findings and interobserver reliability

Background: Magnetic resonance (MR) imaging is frequently used to diagnose arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVC/D). However, the reliability of various MR imaging features for diagnosing ARVC/D is unknown. The purpose of this study was to determine which morphologic MR imaging features have the greatest interobserver reliability for diagnosing ARVC/D. Methods: Forty-five sets of films of cardiac MR images were sent to 8 radiologists and 5 cardiologists with experience in this field. There were 7 cases of definite ARVC/D as defined by the Task Force criteria. Six cases were controls. The remaining 32 cases had MR imaging because of clinical suspicion of ARVC/D. Readers evaluated the images for the presence of (a) right ventricle (RV) enlargement, (b) RV abnormal morphology, (c) left ventricle enlargement, (d) presence of high T1 signal (fat) in the myocardium, and (e) location of high T1 signal (fat) on a Likert scale with formatted responses. Results: Readers indicated that the Task Force ARVC/D cases had significantly more (χ2 = 119.93, d.f. = 10, p < 0.0001) RV chamber size enlargement (58%) than either the suspected ARVC/D (12%) or no ARVC/D (14%) cases. When readers reported the RV chamber size as enlarged they were significantly more likely to report the case as ARVC/D present (χ2= 33.98, d.f. = 1, p < 0.0001). When readers reported the morphology as abnormal they were more likely to diagnose the case as ARVC/D present (χ2 = 78.4, d.f. = 1, p < 0.0001), and the Task Force ARVC/D (47%) cases received significantly more abnormal reports than either suspected ARVC/D (20%) or non-ARVC/D (15%) cases. There was no significant difference between patient groups in the reported presence of high signal intensity (fat) in the RV (χ2 = 0.9, d.f. = 2, p > 0.05). Conclusions: Reviewers found that the size and shape of abnormalities in the RV are key MR imaging discriminates of ARVD. Subsequent protocol development and multicenter trials need to address these parameters. Essential steps in improving accuracy and reducing variability include a standardized acquisition protocol and standardized analysis with dynamic cine review of regional RV function and quantification of RV and left ventricle volumes.

Effect of intravenous contrast media on proximal and distal tubular hydrostatic pressure in the rat kidney.

The effect of i.v. injection of contrast media (CM, 1 600 mg I/kg b.w.) on proximal and distal tubular hydrostatic pressure (PTHP, DTHP) in the rat was investigated using a micropuncture technique. The PTHP and DTHP after injection of diatrizoate, iohexol, ioxaglate, or mannitol returned to control values within approximately 20 min. However, following iotrolan injection PTHP was still elevated above control levels after 35 min while DTHP remained elevated throughout the experiment (50 min). Iotrolan has a lower osmotic potential than the other CM when given in equivalent iodine doses. The concentration of iotrolan may thus increase more along the tubules than the other CM and consequently lead to a higher viscosity of urine, resulting in increases in PTHP and DTHP. The high intratubular pressure induced by iotrolan may explain our previous findings of reduced single nephron glomerular filtration rate caused by this CM.

Effects of Intravenous Contrast Media on Cortical and Medullary Blood Flow in the Rat Kidney

The effect of slow intravenous infusion of contrast medium (CM) (1600 mg I/kg body weight) on cortical blood flow (BF) and medullary BF in rat kidneys was investigated by laser-Doppler flowmetry on either renal cortex or exposed renal papillas (inner medulla). The effect on cortical BF was evaluated after infusion of either ioxaglate, iohexol, or ioxithalamate. Mannitol and Ringers solution were used as control substances. The effect on medullary BF was examined after infusion of either ioxaglate, iohexol, iopamidol, ioxithalamate, or mannitol. BF was measured continuously during a 30-minute control period and a 60-minute experimental period, starting with the CM infusion. Cortical BF was unchanged in the ioxaglate group and significantly increased in the iohexol, ioxithalamate, and mannitol groups (P less than .05). Medullary BF was moderately increased in the ioxaglate group (P less than .05) but moderately decreased in the groups that received iohexol, iopamidol, ioxithalamate, or mannitol (P less than .05). The reduction in medullary BF following infusion of the ratio 3.0 nonionic CM and of the ratio 1.5 ionic CM might be one contributory mechanism to the pathogenesis of CM nephropathy, especially in the presence of microangiopathy in the kidney.

Influence of Contrast Media on Single Nephron Glomerular Filtration Rate in Rat Kidney: A comparison between diatrizoate, iohexol, ioxaglate, and iotrolan

The effects of slow (10 min) i.v. infusions of contrast media (CM, 1 600 mg I/kg b.w.) on single nephron glomerular filtration rate (SNGFR) in the rat kidney were investigated using a micropuncture technique. Diatrizoate, iohexol, or ioxaglate did not change SNGFR, although a tendency towards a transient suppression was seen during the infusion phase. Iotrolan infusion, however, decreased SNGFR (p < 0.05) and the value still remained below the control value 25 min after the start of infusion. Iotrolan is a nonionic dimeric CM and has a lower osmotic effect in the tubules than the ionic dimeric CM and the monomeric CM when given in iodine equivalent doses. These characteristics of iotrolan have probably some influence on the depression of SNGFR after iotrolan injection.

The haemodynamic effects of iodinated water soluble radiographic contrast media : a review

All classes of iodinated water-soluble radiographic contrast media (RCM) are vasoactive with the iso-osmolar dimers inducing the least changes in the vascular tone. The mechanisms responsible for RCM-induced changes in the vascular tone are not fully understood and could be multifactorial. A direct effect on the vascular smooth muscle cells causing alterations in the ion exchanges across the cell membrane is thought to be an important factor in RCM-induced vasodilatation. The release of the endogenous vasoactive mediators adenosine and endothelin may also play a crucial role in the haemodynamic effects of RCM particularly in the kidney. In addition, the effects of RCM on blood rheology can cause a reduction in the blood flow in the microcirculation. The purpose of this review is to discuss the pathophysiology of the haemodynamic effects of RCM and to offer some insight into the biology of the endothelium and vascular smooth muscle cells as well as the pharmacology of the important vasoactive mediators endothelin and adenosine.

Iodine concentrations in the rat kidney measured by x-ray microanalysis Comparison of concentrations and viscosities in the proximal tubules and renal pelvis after intravenous injections of contrast media

Purpose: To measure the iodine concentrations in the proximal tubules and renal pelvis after i.v. injections of contrast media (CM) at 1600 mg I/kg b.w., using a micro-puncture technique and X-ray microanalysis Material and Methods: The correlation between the viscosity of each CM and its iodine concentration was evaluated and the viscosity of the fluid in the proximal tubule and renal pelvis was estimated in rats Results: After iotrolan injection, the iodine concentration in the proximal tubular fluid had increased to values about three times higher than those reached with diatrizoate, iohexol and ioxaglate. Similarly, iotrolan tended to produce a higher iodine concentration in the renal pelvis than did the other CM. the urine viscosity in the renal pelvis was much higher after the iotrolan injection than after the other CM injections Conclusion: High urine viscosity after iotrolan injection can at least partly explain our previous findings of a prolonged increase in tubular hydrostatic pressure and a prolonged decrease in the single nephron glomerular filtration rate after administration of this CM

Effect of furosemide or mannitol before injection of a non-ionic contrast medium on intrarenal oxygen tension.

Oxygen tension (pO2) in rat renal cortex and outer medulla was studied after an intravenous injection of mannitol or furosemide, followed 10 minutes later by an intravenous injection of the non-ionic X-ray contrast medium (CM) iopromide (370 mg iodine/ml). Ten minutes after mannitol injection, before injection of CM, pO2 in the medulla had decreased from a control level of 32 +/- 3 to 28 +/- 4 mm Hg. The addition of CM caused a further decrease, to 24 +/- 5 mm Hg, which was a significant reduction. Ten minutes after furosemide injection the pO2 in the medulla had increased significantly, from a control value of 32 +/- 2 to 44 +/- 4 mm Hg. Injection of CM caused a significant decrease in pO2 to 37 +/- 3 mm. Ringers solution (n = 6) caused no changes. We conclude that pretreatment with mannitol or furosemide does not prevent the CM-induced decrease in pO2 in the outer medulla.

Influence of Contrast Media on Single Nephron Glomerular Filtration Rate in Rat Kidney

The effects of slow (10 min) i.v. infusions of contrast media (CM, 1 600 mg I/kg b.w.) on single nephron glomerular filtration rate (SNGFR) in the rat kidney were investigated using a micropuncture technique. Diatrizoate, iohexol, or ioxaglate did not change SNGFR, although a tendency towards a transient suppression was seen during the infusion phase. Iotrolan infusion, however, decreased SNGFR (p<0.05) and the value still remained below the control value 25 min after the start of infusion. Iotrolan is a nonionic dimeric CM and has a lower osmotic effect in the tubules than the ionic dimeric CM and the monomeric CM when given in iodine equivalent doses. These characteristics of iotrolan have probably some influence on the depression of SNGFR after iotrolan injection.

Effects of High- and Low-Osmolar Contrast Media on Renal Plasma Flow and Glomerular Filtration Rate in Euvolaemic and Dehydrated Rats: A Comparison between Ioxithalamate, Iopamidol, Iohexol and Ioxaglate

The effects of a slow intravenous injection of contrast media (CM) on renal function and haemodynamics were investigated in euvolaemic and dehydrated rats. Iodine-equivalent doses (1600 mg I/kg body weight) of ioxithalamate, ioxaglate, iopamidol and iohexol were used. Glomerular filtration rate (GFR) and renal plasma flow (RPF) were assessed with clearance techniques. In euvolaemic rats no statistically significant decrease in GFR or RPF was found after CM injections. In the dehydrated rats the changes in GFR were more pronounced and this was significantly decreased in the ioxithalamate and iopamidol groups while RPF was still not decreased. This study indicates that dehydration potentiates adverse effects of CM on GFR and that there may be differences between the effects of low-osmolar and high-osmolar CM on GFR and also between different low-osmolar CM.