General colorimetric method for DNA diagnostics allowing direct solid-phase genomic sequencing of the positive samples.

A system for rapid colorimetric detection of specific genome DNA fragments amplified by the polymerase chain reaction (PCR) is described that has been designed to allow direct solid-phase sequencing of positive samples. The amplified material is immobilized on magnetic beads by using the biotin streptavidin system. An Escherichia coli lac operator DNA sequence is incorporated in the amplified material during the second step of a nested primer procedure.

Rapid detection and sequencing of specific in vitro amplified DNA sequences using solid phase methods.

We describe a rapid solid phase assay for detection and sequencing of DNA sequences based on selective introduction of biotin and isotope into the specific DNA fragment amplified by the polymerase chain reaction (PCR). A two-step PCR procedure is used to lower the background signal. The in vitro amplified material is immobilized on magnetic beads with covalently coupled streptavidin and the amount of bound label is measured.

Rapid colorimetric detection of in vitro amplified DNA sequences.

A colorimetric assay to detect immobilized amplified nucleic acids has been designed. This approach provides a rapid assay, suitable for clinical diagnosis, to analyze DNA sequences amplified by the polymerase chain reaction. The specific DNA sequences are captured on a solid support by the use of a recombinant fusion protein consisting of the Escherichia coli lac repressor and staphylococcal protein A.

Affinity purification of specific DNA fragments using a lac repressor fusion protein.

A new method for purification of specific DNA sequences using a solid phase technique has been developed based on a fusion between the Escherichia coli lac repressor gene (lacI) and the staphylococcal protein A gene (spa). The fusion protein, expressed in Escherichia coli, is active both in vivo and in vitro with respect to its three functional activities (DNA binding, IPTG induction, and IgG binding). The recombinant protein can be immobilized in a one-step procedure with high yield and purity using the specific interaction between protein A and the Fc-part of immunoglobulin G.

Contact allergy to Compositae plants in patients with summer-exacerbated dermatitis.

16 patients with summer-exacerbated dermatitis were examined by patch testing and photopatch testing with a battery of Compositae and standard allergens. IgE and RAST, and the thresholds to UVA and UVB, were determined. 6 female and 2 male patients showed allergic contact reactions to one or more Compositae extracts from flowers common in Sweden, also used in skin care products.

Nickel chloride or nickel sulfate? Irritation from patch-test preparations as assessed by laser Doppler flowmetry.

To obtain an optimal test preparation for diagnosing nickel contact sensitivity, one can vary the nickel salt, vehicle and concentration, length of exposure, patches, and tape. Nickel chloride has been suggested as an alternative to nickel sulfate in the standard tray. However, a sensitive and noninvasive method of recording skin blood flow (laser Doppler flowmetry) showed in healthy subjects that the chloride was more irritating to the skin than the sulfate when petrolatum was used as vehicle.

Experience with the Hemasite device in haemodialysis and haemofiltration patients with vascular access problems.

Multiple vascular access failure is a frequent problem and we report our experience with 32 Hemasite devices implanted in 28 patients with end-stage renal disease. All of the patients were on maintenance haemodialysis or haemofiltration with a range of treatment of 1-17 years. Their mean age was 58 +/- 14 years (range 28-72 years).

Influence of the age of cadaveric kidney donors on transplantation outcome and rate of surgical complications.

Cadaveric kidney graft recipients, treated according to a strict, high dose CyA protocol, were followed prospectively for one year. The aim was to study the impact of donor age on transplantation outcome in a homogenously immunosuppressed patient material. The patients were divided into 2 groups; G1: donor age less than or equal to 50 years (mean 34.

Platelet-derived growth factor receptors in the kidney--upregulated expression in inflammation.

Major features of a long-standing inflammation in the kidney are vascular proliferation, glomerulosclerosis, interstitial fibrosis and tubular atrophy, leading to a gradual deterioration of the renal function. In this study we have investigated the expression of B-type receptors for platelet-derived growth factor (PDGF) in frozen sections from normal and inflamed kidneys. Immunohistochemical techniques, employing two monoclonal antibodies specific for PDGF B-type receptors, were used.

The heterodimeric association between the membrane proteins of Semliki Forest virus changes its sensitivity to low pH during virus maturation.

The budding and the fusion processes of the enveloped animal virus Semliki Forest virus serve the purpose of transporting its nucleocapsid, containing its genome, from the cytoplasm of an infected cell into that of an uninfected one. We show here that, in the infected cell, the viral membrane (spike) proteins p62 and E1 are organized as heterodimers which are very resistant to dissociation in acidic conditions. In contrast, the mature form of the heterodimer, E2E1, which is found in the virus particle and which is generated by proteolytic processing of p62, is very prone to dissociate upon treatment with mildly acidic buffers.

Preservation-induced pancreatitis in an isolated perfused pancreas model in the dog.

In order to investigate pancreatitis caused by cold ischemic damage to pancreatic grafts, an isolated, normothermic, ex vivo perfusion model was employed. Canine pancreases were subjected to 24 and 48 h of cold ischemia and then reperfused. The results showed that cold ischemia results in pancreatitis as measured by weight gain (tissue edema) and elevated leakage of amylase into the perfusate.

Relevance of additive components of University of Wisconsin cold-storage solution. An experimental study in the rat.

The purpose of this study was to evaluate the value of additive components and colloid included in the University of Wisconsin (UW) solution. Therefore, this solution was compared with a solution consisting of the basic components of the UW solution (potassium lactobionate, raffinose, phosphate buffer and MgSO4). We employed a method of measuring the amount of chromium-51-labeled erythrocyte trapped in the medullary vasculature 20 min after reperfusion of kidney grafts cold-stored for 24-48 h in either the basic UW (bUW) or the original UW (oUW) solution.

Influence of different preservation solutions and intentional hemodilution of recipient on viability of preserved and transplanted rat kidneys.

A total of 81 rat kidney grafts, flushed out and cold stored in either Sacks' or University of Wisconsin (UW) solution, were transplanted into hemodiluted (Hct = 30% +/- 4%) or untreated (Hct = 43% +/- 3%) recipients. The cold ischemia times (CIT) used were 24 and 36 h. One week after transplantation, the surviving recipients (n = 67) were contralaterally nephrectomized.

Percutaneous absorption of 3 organic solvents in the guinea pig (I). Effect of physical and chemical injuries to the skin.

The influence of injuries on the percutaneous absorption of 3 organic solvents (butanol, toluene, 1,1,1-trichloroethane) was studied in the guinea pig. Acute (stripping, sandpaper abrasion, needle abrasion, delipidization) and subacute (irritant and allergic contact dermatitis) injuries were induced by physical or chemical treatment of the skin. Absorption of solvents was quantified by blood analysis.

The effect of type of preservation solution and hemodilution of the recipient on postischemic erythrocyte trapping in kidney grafts. An experimental study in the rat.

The role of recipient hemodilution for postischemic renal medullary red cell trapping was investigated after different periods of cold storage in a conventional cold storage solution (Sacks'). At all cold storage times investigated (4, 12, 24, and 48 hr) medullary red cell trapping was reduced by isovolemic hemodilution, with about 50% reduction of recipient hematocrit. Trapping was also reduced when a modification of a new preservation solution (University of Wisconsin solution [UW]) was used and compared with flush-out and storage in a standard preservation solution (Sacks').

Continuous arteriovenous hemofiltration in the treatment of 100 critically ill patients with acute renal failure: report on clinical outcome and nutritional aspects.

The clinical outcome for 100 consecutive patients with multiorgan failure including acute renal failure (ARF) was studied. Fifty-eight of the patients had acute renal failure due to complications during and after major surgery. Seventy-three of the patients had a urine output of less than 400 ml/24 hours.

Organ flush out solutions and cold storage preservation solutions: effect on organ cooling and post ischemic erythrocyte trapping in kidney grafts. An experimental study in the rat.

Three flush out and cold storage solutions were tested for flow rate during flush out, ability to decrease organ temperature, degree of tissue edema and degree of preservation damage of the kidney as measured by the amount of post transplantation erythrocyte trapping in the renal medulla. The solutions tested were a modification of a new perfusion solution (University of Wisconsin; mUW), a standard preservation solution (Sacks') and an extracellular histidine solution (Frödin-Wolgast; FW). The flow rate was significantly higher for FW compared to mUW and Sacks' and consequently a more rapid decrease of organ temperature was achieved.

Outcome of continuous arteriovenous haemofiltration (CAVH) in one centre.

Continuous arteriovenous haemofiltration (CAVH) has been adopted as the treatment of choice for acute renal failure (ARF) in critically ill patients in the intensive care units of Uppsala University Hospital since 1982. To know the outcome of CAVH during the last one and a half year this retrospective study was done on those patients seen in July 1987-December 1988. Forty patients aged 2 months-84 years (mean 57 years) were included.