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Biomedical subjects

A Holmberg

Publications and source records attributed to A Holmberg.

At least 19 recordsLinked to original sources

High levels of hepatitis B virus DNA in body fluids from chronic carriers.

Chronic infection with hepatitis B virus (HBV) is a major global health problem. Transmission is mainly blood-borne, although the route of infection during horizontal transmission in childhood is unclear. Nosocomial outbreaks of HBV have been widely reported, but have mainly focused on blood-borne transmission. There is limited knowledge of the viral load levels in other body fluids. In the present study, chronic HBV carriers were tested for the presence of HBV DNA in serum, saliva, nasopharyngeal fluid, urine and tears by means of qualitative and quantitative polymerase chain reaction (PCR) methods. Twenty-five patients who were positive for HBV DNA with both PCRs were included. Low titres in real-time PCR corresponded with weak bands in the qualitative assay. HBV DNA was found in two urine samples, 10 saliva samples, five nasopharyngeal swabs and in tear fluid from four patients. One highly viraemic HBeAg-positive carrier with serum HBV DNA levels of 7 x 10(9) genome copies had high copy numbers detected in both saliva and nasopharyngeal fluid. These results demonstrate that highly viraemic HBV carriers may have high titres of HBV DNA in other body fluids. This has particular importance for infection control programmes and regulations, underlining the importance of aiming towards regular HBV DNA testing and thus infectivity assessment of chronic carriers in order to prevent transmission.

Adult↗

Molecularly imprinted polymers for the determination of a pharmaceutical development compound in plasma using 96-well MISPE technology.

The use of molecularly imprinted polymers (MIPs) as sorbents for the solid phase extraction (SPE) of a pharmaceutical compound in development, prior to quantitative analysis was investigated. Three MIPs were synthesised using a structural analogue as the template molecule. Each polymer was prepared with different monomers and porogens. The MIPs were then tested for their performance both in organic and aqueous environments, the final aim being to load plasma directly onto the polymers. At an early development stage, there is a limited amount of compound available. Due to this limitation, reducing the amount of template required for imprinting was investigated. A MIP capable of extracting the analyte directly from plasma was produced. The specificity of the polymer allowed the method to be validated at a lower sensitivity than a more conventional SPE assay. For the first time, MIPs were packed into 96-well blocks enabling high throughput analysis. The analytical method was fully validated for imprecision and inaccuracy down to 4 ng/ml in plasma.

Drug Design↗

Activation of coagulation and fibrinolytic systems in patients with CLI is not normalized after surgical revascularisation.

OBJECTIVE: To study the activation of coagulation and fibrinolysis before, during and after surgical revascularisation in patients with critical limb ischemia (CLI). DESIGN: Prospective clinical study. MATERIALS AND METHODS: Forty patients with CLI underwent femoro-popliteal or femoro-distal reconstruction and were compared to a control-group. Measurements of prothrombin-fragment 1+2 (F1+2) and thrombin-antithrombin complex (TAT) assessed activation of coagulation. Fibrinolysis was determined by tissue plasminogen activator (tPA), plasminogen activator inhibitor (PAI-1) and fibrin degradation product (D-dimer). The inflammatory mediators: Interleukin 2 receptor (IL-2-rec), Interleukin 6 (IL-6), Interleukin 10 (IL-10) and Monocyte chemoattractant protein 1 (MCP-1) was also analysed. RESULTS: Patients (in 35 of the 40 reconstruction was possible) were operated upon using either vein (n=23) or ePTFE (n=12) grafts. Patients with CLI had a preoperative prothrombotic state as indicated by high TAT-levels and also ongoing fibrinolysis with high levels of t-PA and D-dimer. After reperfusion an ongoing prothrombotic state for the first week was demonstrated. A significant as well as defective fibrinolysis was also seen with increased levels of tPA and D-dimer unopposed by PAI-1 after one week and also after 30 days. Increased levels of inflammatory mediators IL-6, IL-10 and MCP-1 was observed after reperfusion and normalised after 30 days. CONCLUSION: This study demonstrates significant disturbances of both the coagulation and fibrinolytic systems before, during and after revascularisation for CLI. This was accompanied by release of inflammatory mediators. A prothrombotic state and increased fibrinolysis were evident also 30 days after successful revascularisation.

Aged↗

Wavelet-based image restoration for compact X-ray microscopy.

Compact water-window X-ray microscopy with short exposure times will always be limited on photons owing to sources of limited power in combination with low-efficency X-ray optics. Thus, it is important to investigate methods for improving the signal-to-noise ratio in the images. We show that a wavelet-based denoising procedure significantly improves the quality and contrast in compact X-ray microscopy images. A non-decimated, discrete wavelet transform (DWT) is applied to original, noisy images. After applying a thresholding procedure to the finest scales of the DWT, by setting to zero all wavelet coefficients of magnitude below a prescribed value, the inverse DWT to the thresholded DWT produces denoised images. It is concluded that the denoising procedure has potential to reduce the exposure time by a factor of 2 without loss of relevant image information.

Algorithms↗

Physical activity in relation to aerobic fitness and body fat in 14- to 15-year-old boys and girls.

The purpose of this study was to examine the strength of the relationship between different variables of physical activity and aerobic fitness and body fat in adolescent boys and girls. Activity energy expenditure (AEE), time spent in a sedentary state, and time spent engaged in moderate and vigorous physical activity (MVPA, > or = 50% peak oxygen uptake, VO2peak) were assessed by the minute-by-minute heart rate monitoring method in 82 randomly selected 14- to 15-year olds (42 boys, 40 girls). Body fat was determined by measuring skinfold thicknesses. VO2peak was measured by indirect calorimetry. Somatic maturity level was determined by percentages of adult (i.e. 18 years) height attained at examination. AEE was related to aerobic fitness for both genders (boys, r = 0.30, P = 0.056; girls, r = 0.45, P = 0.003). For boys, there was a significant relationship between maturity level and VO2peak (r = 0.48, P < 0.001). For both genders, body fat was significantly and negatively related to VO2peak (r = -0.48 and r = -0.43, P < 0.01). Body fat and maturity explained 47% of the variation in VO2peak in boys, whereas AEE and body fat explained 22% of the variation in VO2peak in girls. No significant associations between physical activity variables and the data on body fat were observed. The total amount of physical activity (AEE) was related to VO2peak, at least in adolescent girls. Although VO2peak seems to be influenced by the maturity level in adolescent boys, the data support the promotion of a daily active lifestyle among young people.

Adipose Tissue↗

Occurrence of neurotrophin receptors and transmitters in the developing Xenopus gut.

The ontogeny of gut innervation in the anuran amphibian Xenopus laevis was studied using immunohistochemistry on sections of whole larvae from NF stages 38-52. Immunoreactivity to acetylated tubulin confirmed the presence of nerve fibres as early as stages 38-39. Actin immunoreactivity was found at stage 41, indicating the presence of smooth muscle cells. Trk-like neurotrophin receptors were occasionally found in nerve fibres as soon as stages 38-39. Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP) immunoreactivities coexisted in nerves innervating the gut wall from stages 40-41, and nitric oxide synthase (NOS) from stage 42. Substance P/neurokinin A (SP/NKA) occurred at stage 42. In all these cases, the first fibres were observed in the oesophagus. Calcitonin gene-related peptide (CGRP) was first observed in nerves at stage 48. In general, VIP/PACAP and NOS innervation was denser than the tachykinin innervation. In conclusion, the development of nerve fibres in the Xenopus gut is probably dependent on neurotrophins that may act via Trk-like receptors and occur before the gut wall is fully organised morphologically. Feeding in Xenopus larvae starts at NF stage 45. The study demonstrates that several of the transmitters investigated are expressed in the gut innervation (and in endocrine cells) prior to this stage.

Animals↗

Gadolinium-enhanced magnetic resonance angiography, digital subtraction angiography and duplex of the iliac arteries compared with intra-arterial pressure gradient measurements.

PURPOSE: to compare gadolinium-enhanced magnetic resonance angiography (Gd-MRA), digital subtraction angiography (DSA) and duplex of the iliac arteries with intra-arterial pressure gradient measurement as the reference method. MATERIALS AND METHODS: Gd-MRA, DSA and duplex examinations of the iliac arteries were performed in 30 patients (60 arteries) with lower-limb arterial occlusive disease. In 29 arteries, pressure measurements were made (n=25) or the artery was found to be occluded on catheterisation (n=4). An aortofemoral peak systolic pressure gradient of 20 mmHg or more was regarded as haemodynamically significant. Stenoses with a diameter reduction of 50% or more on MRA or DSA, or an increase in peak systolic velocity greater than 150% (duplex) were considered significant. MRA examinations were evaluated by means of maximum intensity projections (MRA-MIP) and using source images and curved multiplanar reconstruction (MRA-MPR). RESULTS: the sensitivity (specificity) for a significant iliac artery stenosis were 81% (75%) for MRA-MIP, 76% (75%) for MRA-MPR, 86% (88%) for DSA, and 72% (88%) for duplex. CONCLUSION: with intra-arterial pressure measurements as the reference method, similar results were achieved with Gd-MRA, DSA and duplex concerning the detection of haemodynamically significant iliac artery stenoses. The use of source images and multiplanar reconstructions resulted in higher accuracy for the detection of occlusions.

Aged↗

Hemorheologic variables in critical limb ischemia before and after infrainguinal reconstruction.

PURPOSE: Plasma and whole blood viscosity are elevated in patients with intermittent claudication. The objectives of this study were to investigate whether critical limb ischemia influences hemorheologic variables and whether the rheologic variables in blood from the affected limb differ from the general circulation. We also intended to study whether successful infrainguinal reconstruction improved hemorheologic variables. METHODS: Ten consecutive patients with critical limb ischemia (CLI) underwent arterial reconstruction, one patient with profundaplasty and nine patients with bypass procedures. Venous blood was sampled from the antecubital vein (arm) and the femoral vein (leg) of the affected limb 1 day before and 1 month after surgery. Ten control subjects (matched according to age, sex, diabetic status, and renal insufficiency) were also sampled. Whole blood viscosity, plasma viscosity, erythrocyte aggregation tendency, and erythrocyte fluidity (the latter variable describing the deformability of the erythrocytes) were measured by means of rotational viscometry. Erythrocyte volume fraction was also determined. Fibrinogen was measured in the patients with CLI. RESULTS: Erythrocyte fluidity, blood viscosity, and erythrocyte volume fraction were lower in patients with CLI than in control subjects (P <.01, P <.01, and P <.05, respectively). No major differences between cubital and femoral vein blood were seen before or after the operation in patients with CLI or in control subjects. Successful revascularization did not influence the hemorheologic variables, except for a decrease in blood viscosity in femoral vein blood (P <.05). CONCLUSION: Hemorheologic properties was impaired in patients with CLI. Because no differences were seen between the systemic and local circulation and because no major improvement occurred 1 month after arterial reconstruction, other mechanisms besides local tissue ischemia may play a role.

Aged↗

Factors affecting motivation to treatment in severely dependent alcoholics.

OBJECTIVE: This article explores environmental, developmental and personality-related factors as predictors of motivation to treatment for alcoholism, using such concepts as: social situation on admission, treatment career, age, education and adventurous patterns of behavior. METHOD: The original population consists of 603 severely dependent alcoholic patients (76% men, 42% compulsorily committed) with a mean age of 40 years. In structural equation model estimation, 85% of the original cases were used (N = 511), made up of compulsorily as well as voluntarily admitted patients. A reduced model was tried on voluntary patients alone. RESULTS: In the main model, patterns of behavior and age were not related to the motivation to treatment, while treatment career had some negative impact. Social problems were related to less motivation to treatment. In turn, a worse social situation was related to more treatment experience. The reduced model on voluntary patients alone confirmed the main findings, although more previous treatment tended to be related to more motivation. CONCLUSIONS: Motivation was mostly related to a better social situation (i.e., having "more to lose"). A collapse in the social situation was more of an obstacle than a promoter of motivation. There was no support for statements that "maturing out" would result in more motivation, and findings were inconclusive concerning the impact on motivation of having had many previous treatment experiences. The pattern of behavior was not directly related to the level of motivation. Based on these findings, the relevance of social work for motivation to treatment is discussed.

Adult↗

Coagulation and fibrinolysis after open infrarenal abdominal aortic aneurysm repair in a long-term perspective.

In patients with abdominal aortic aneurysms (AAA) the coagulation and fibrinolytic systems have been found to be activated preoperatively. Does the increased activity of the coagulation and fibrinolytic systems persist after AAA surgery in a long-term perspective? Prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT), tissue plasminogen activator (tPA), human plasminogen activator inhibitor type 1, and human cross-linked fibrin degradation product (D-dimer) were analysed in 18 patients after open AAA surgery (postop-AAA). The median time between surgery and blood sampling was 19 months (range, 5-37 months). Comparisons were made with both preoperative values of 23 patients with AAA (preop-AAA) as well as 20 age-matched healthy controls (AMC). F1+2, TAT, and D-dimer in preop-AAA were significantly higher compared to AMC (p<0.001). In post-op AAA patients these parameters were significantly lower compared to preop-AAA (p<0.05 for F1+2 and TAT, p<0.001 for D-dimer). However, TAT and D-dimer levels were still higher in postop-AAA than in AMC (p<0.01 for both). The activity of the coagulation and fibrinolytic systems seems to decrease after AAA surgery. However, the activity is still higher than in healthy AMC. A possible explanation may be that the thrombogenicity is lower in a vascular graft than in an aneurysmal sac but still higher than in a nonaneurysmal aorta.

Aged↗

Truncated fragments in polymerase chain reaction-based DNA sequencing.

Reconstitution experiments were performed by using an ordinary dye-primer protocol of a template spiked with known amounts of truncated fragments. We observed that as little as 0.2 mole-percentage of the truncated fragment caused sequence interpretation problems. Two protocols were developed for sequencing with dye-labeled terminators; this eliminates the problems with truncated fragments, which are adapted to a one-dye chemistry. One was designed for single extension sequencing using T7 DNA polymerase and one for cycle sequencing. To avoid precipitation and centrifugation and to facilitate automation, the dye-terminator protocols included the use of a biotinylated sequencing primer. Thus, the Sanger fragments were recovered and, by magnetic separation, washed and released by formamide, EDTA, and heat treatment before loading on the electrophoresis gel. Integrated procedures for sequencing PCR products using one-dye-labeled terminators suitable for automation are described. High quality data in terms of long reads and detection of polymorphisms is obtained. The protocols serve as attractive alternatives to internal labeling and dye-primer approaches.

Base Sequence↗

Selection analysis of single-stranded clones using solid-phase techniques.

An efficient selection analysis of single-stranded DNA fragments using subtractive hybridization on magnetic beads and fluorescence-based fragment analysis is described. This approach facilitates either direct cloning of captured fragments or removal of common clones from cDNA or shot-gun clone libraries.

Animals↗

Cytokine and fibrinogen response in patients undergoing open abdominal aortic aneurysm surgery.

OBJECTIVES: To assess whether open abdominal aortic aneurysm (AAA) surgery influences cytokines and fibrinogen. METHODS: Twenty-three consecutive patients operated on for AAA were compared to 11 operated controls and 20 age-matched controls. Cubital blood was sampled pre-, intra- and postoperatively and femoral blood also sampled intraoperatively. RESULTS: Preoperatively, interleukin (Il)-6 was elevated in AAA patients. During aortic clamping, Il-6, Il-10 and monocyte chemoattractant protein-1 (MCP-1) increased significantly (p < 0.001, p < 0.01 and p < 0.05 respectively) while soluble interleukin-2 receptor (sIl-2R) and fibrinogen decreased significantly (p < 0.001 for both). After aortic declamping, Il-6, Il-10 and MCP-1 had further significant increases compared with levels during aortic clamping while sIl-2R had a further non-significant and fibrinogen a significant decrease (p < 0.05 in cubital and p < 0.001 in femoral blood). One week postoperatively Il-6, Il-10 and MCP-1 had all decreased but were still significantly elevated compared with baseline values while sIl-2R and fibrinogen showed an increase in comparison with baseline (p < 0.001 for both). Intraoperative levels of Il-6 and Il-10 showed a significant co-variation with the magnitude of operative trauma. CONCLUSIONS: These data indicate that open AAA surgery induces a profound inflammatory and coagulative response which persists at one week postoperatively.

Aged↗

Ischaemia and reperfusion during open abdominal aortic aneurysm surgery induce extensive thrombin generation and activity.

OBJECTIVES: does open surgery for abdominal aortic aneurysm (AAA) influence coagulation? METHODS: in 23 patients operated on for AAA, cubital blood was sampled pre-, intra- and postoperatively. Femoral blood was also sampled intraoperatively. RESULTS: preoperatively, prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT) and soluble fibrin (SF) were elevated in AAA patients. During aortic clamping all parameters increased significantly in cubital blood (p<0.01) as well as in femoral blood (p<0.001) and after aortic declamping F1+2 and TAT increased further. F1+2, TAT and SF were significantly higher in femoral than cubital blood. Postoperatively F1+2 and TAT returned to preoperative values, while SF still had a significantly higher level than preoperatively (p<0.001). Blood loss showed co-variation with F1+2 increase in femoral blood after aortic declamping (p<0.05). CONCLUSIONS: these data indicate that the coagulation system was strongly activated by the occurrence of an AAA. During AAA surgery a further extensive activation was seen. The activity was still high, but on decline, one week postoperatively. Ischaemia and reperfusion of the lower part of the body were the major stimuli for thrombin generation and activity.

Adult↗

Gene discovery in the wood-forming tissues of poplar: analysis of 5, 692 expressed sequence tags.

A rapidly growing area of genome research is the generation of expressed sequence tags (ESTs) in which large numbers of randomly selected cDNA clones are partially sequenced. The collection of ESTs reflects the level and complexity of gene expression in the sampled tissue. To date, the majority of plant ESTs are from nonwoody plants such as Arabidopsis, Brassica, maize, and rice. Here, we present a large-scale production of ESTs from the wood-forming tissues of two poplars, Populus tremula L. x tremuloides Michx. and Populus trichocarpa 'Trichobel.' The 5,692 ESTs analyzed represented a total of 3,719 unique transcripts for the two cDNA libraries. Putative functions could be assigned to 2,245 of these transcripts that corresponded to 820 protein functions. Of specific interest to forest biotechnology are the 4% of ESTs involved in various processes of cell wall formation, such as lignin and cellulose synthesis, 5% similar to developmental regulators and members of known signal transduction pathways, and 2% involved in hormone biosynthesis. An additional 12% of the ESTs showed no significant similarity to any other DNA or protein sequences in existing databases. The absence of these sequences from public databases may indicate a specific role for these proteins in wood formation. The cDNA libraries and the accompanying database are valuable resources for forest research directed toward understanding the genetic control of wood formation and future endeavors to modify wood and fiber properties for industrial use.

Arabidopsis↗

Expression of epidermal growth factor receptor in urinary bladder cancer metastases.

Bladder cancers frequently exhibit an increased number of epidermal growth factor receptors (EGFR) in comparison to normal urothelium. The EGFR could potentially be a target for toxic conjugates. The aim of our study was to compare the expression of EGFR in metastases with concurrent or primary tumour in the urinary bladder using immunohistochemical techniques and a monoclonal antibody. Tumour material from 20 patients was investigated. The majority (13/20) of the metastases were homogeneously stained and showed a moderate to strong membranous staining for EGFR. The expression of EGFR in primary bladder tumours and metastases was similar. There was no indication that tumour tissue exposed to chemotherapy or radiation had a decreased number of EGFR. Targeting of the EGFR thus seems potentially applicable to metastatic disease.

Aged↗

Direct sequencing of bacterial artificial chromosomes (BACs) and prokaryotic genomes by biotin-capture PCR.

Determination of unknown DNA sequences adjacent to known segments is an important task in genome-related research. We have applied the methodology of biotin-capture PCR for direct sequencing of bacterial artificial chromosomes (BACs) and bacterial genomes. The strategy involves extension of a biotinylated primer from a known locus into unknown regions of the template to yield single-stranded DNA, which is immobilised onto paramagnetic beads. An arbitrary primer initiates extension from the unknown region and back towards the known locus. The arbitrary primer contains a universal primer 'handle', which is utilised for subsequent amplification. The PCR products are then directly sequenced by solid-phase or cycle sequencing. The fact that BACs or bacterial chromosomes can be sequenced without prior purification or subcloning might be useful in numerous applications, such as gap-filling, sequencing of regulatory regions upstream known genes and determination of intron/exon-boundaries.

Base Sequence↗