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PubMed2002

A simple method for serving Web hypermaps with dynamic database drill-down.

BACKGROUND: HealthCyberMap http://healthcybermap.semanticweb.org aims at mapping parts of health information cyberspace in novel ways to deliver a semantically superior user experience. This is achieved through "intelligent" categorisation and interactive hypermedia visualisation of health resources using metadata, clinical codes and GIS. HealthCyberMap is an ArcView 3.1 project. WebView, the Internet extension to ArcView, publishes HealthCyberMap ArcView Views as Web client-side imagemaps. The basic WebView set-up does not support any GIS database connection, and published Web maps become disconnected from the original project. A dedicated Internet map server would be the best way to serve HealthCyberMap database-driven interactive Web maps, but is an expensive and complex solution to acquire, run and maintain. This paper describes HealthCyberMap simple, low-cost method for "patching" WebView to serve hypermaps with dynamic database drill-down functionality on the Web. RESULTS: The proposed solution is currently used for publishing HealthCyberMap GIS-generated navigational information maps on the Web while maintaining their links with the underlying resource metadata base. CONCLUSION: The authors believe their map serving approach as adopted in HealthCyberMap has been very successful, especially in cases when only map attribute data change without a corresponding effect on map appearance. It should be also possible to use the same solution to publish other interactive GIS-driven maps on the Web, e.g., maps of real world health problems.

Journal Article↗
PubMed1998

Nucleotide-dependent structural changes in dimeric NCD molecules complexed to microtubules.

Complexes consisting of motor domains of the kinesin-like protein ncd bound to reassembled brain microtubules were visualised using cryoelectron microscopy and helical image reconstruction. Different nucleotide-associated states of a dimeric construct (NDelta295-700) of ncd were analysed to reveal ADP-containing, AMP.PNP-containing and empty (rigor) conformations. In these three states, each thought to mimic a different stage in ATP turnover, the double-headed motors attach to the microtubules by one head only, with the free head tethered in relatively fixed positions. The three structures differ both in the way the attached heads interact with tubulin and in the position of the tethered heads. In the strongly binding rigor and AMP.PNP (ATP-like) states, the attached head makes close contact with both subunits of a tubulin heterodimer. In the weakly bound ADP state, the contact made by the attached head with the monomer closer to the plus end appears to be more loose. Also, in the ATP-like state, the free head tilts nearer to the plus end than in the other two states. The data argue against model mechanisms in which a conformational change in the bound head guides the free head closer to its next binding site; on the contrary, the transition from ADP-filled via rigor to the AMP.PNP (ATP-like) state of the bound head produces a small motion of the free head in the counter-productive direction. However, the observation that the tethered head points towards the minus end, in all three states, is consistent with the idea that the relative arrangement of the heads in a dimer is a major determinant of directionality.

Adenosine Diphosphate↗
PubMed1999

Scintigraphy for evaluating early aspiration after oral feeding in patients receiving prolonged ventilation via tracheostomy.

OBJECTIVE: In tracheotomised patients the incidence of aspiration is difficult to determine because investigators often apply different criteria. In this study a scintigraphic method was used to visualise feeding aspiration directly and the results were compared with clinical evidence of aspiration. DESIGN: Prospective study in difficult-to-wean patients with tracheostomy. SETTING: Respiratory ICU. PATIENTS AND METHODS: The study population consisted of 62 consecutive patients (16 females, age: 64.1+/-11.1 years). All patients were tracheotomised and had previously been long-term ventilated in other ICUs due to weaning failure. The scintigraphic test was performed during spontaneous breathing. The standard nutrition consisted of a liquid, semi-liquid and solid meal which was labelled with 100 MBq 99mTc-human serum albumin. MEASUREMENTS AND RESULTS: Scintigraphic aspiration (SA) was defined as positive if radioactivity was detected in the bronchial system using a scintillation camera. Furthermore, aspiration was proven clinically (CA). CA and SA yielded identical results in 54 of the 62 patients [10 positive (16%) and in 44 negative (71%)]. CA, but not SA, was seen in 4/62 (6.5%) and SA, but not CA, was found in 4/62 (6.5%) patients. CONCLUSIONS: Our data re-emphasise that aspiration in tracheotomised patients is common (in our study approximately 30%). The scintigraphic method failed to identify all tracheotomised patients with clinically significant aspiration; however, it did suggest that some patients had subclinical aspiration.

Aged↗
PubMed1995

Theileria annulata sporozoite antigen fused to hepatitis B core antigen used in a vaccination trial.

A C terminal fragment (SR1) of SPAG-1, a sporozoite surface antigen of Theileria annulata, has been expressed as a fusion protein in the e1 loop of hepatitis B core antigen (HBcAg). This recombinant antigen (HBcAg-SR1) is produced in the form of self-assembling polyhedral particles which have been visualised under the electron microscope. Cattle immunised with HBcAg-SR1 produced high titres of neutralising antibodies. A significant T cell response to both the HBcAg and SR1 determinants was observed but evidence of a T suppressor determinant in SR1 was also revealed. Immunised cattle showed some evidence of protection to sporozoite challenge as assessed by severity of the disease. The significance of these findings for the development of a sub-unit vaccine against T. annulata is discussed.

Animals↗
PubMed1997

Iopentol (Imagopaque 250) compared with diatrizoate (Urografin 219) in endoscopic retrograde cholangio-pancreatography (ERCP). A clinical trial assessing safety (adverse events and S-pancreatic iso-amylase) and diagnostic information (VAS).

The efficacy and safety of the non-ionic contrast medium iopentol, 250 mg I/ml (Imagopaque, Nycomed Imaging AS, Oslo, Norway) were evaluated and compared to those of the ionic contrast medium diatrizoate 219 mg I/ml (Urografin, Schering AG, Berlin, Germany). The trial was carried out as a randomized, double-blind comparative two-group study. One hundred and sixteen patients completed the study: 59 received iopentol and 57 received diatrizoate, the contrast medium used according to routine hospital procedure for ERCP. Demographic data and details of the ERCP procedure were comparable for the two contrast medium groups. Safety was assessed by monitoring serum pancreatic iso-amylase and by recording adverse events. Efficacy was evaluated through assessment of the diagnostic information and the quality of radiographs. Twenty-one of the patients receiving iopentol and 27 of the patients receiving diatrizoate reported adverse events during the ERCP procedure. Pain was the kind of procedure-related event most frequently reported. Three patients experienced serious adverse events with a fatal outcome 2, 10 and 12 days after the ERCP procedure, respectively. The principal investigator concluded for all three serious adverse events that any causal relationship with the contrast medium injected was unlikely. A mean change in serum pancreatic iso-amylase after contrast medium injection was noted in both contrast medium groups. However, statistical analysis did not show any significant difference between mean changes for the two groups. The efficacy results, both in terms of diagnostic information and quality of radiographic visualisation, were comparable for the two contrast media. In conclusion, the study did not show any statistically significant differences between the two contrast media as regards safety or efficacy in ERCP.

Amylases↗
PubMed2004

CMP-sialic acid synthetase of the nucleus.

Sialic acids of cell surface glycoconjugates play a pivotal role in the structure and function of animal cells and in some bacterial pathogens. The pattern of cell surface sialylation is species specific, and, in the animal, highly regulated during embryonic development. A prerequisite for the synthesis of sialylated glycoconjugates is the availability of the activated sugar-nucleotide cytidine 5'-monophosphate N-acetylneuraminic acid (CMP-NeuAc), which provides the substrate for sialyltransferases. Trials to purify the enzymatic activity responsible for the synthesis of CMP-NeuAc from different animal sources demonstrated that the major localisation of the enzyme is the cell nucleus. These earlier findings were confirmed when the murine CMP-NeuAc synthetase was cloned and the subcellular transport of recombinant epitope tagged forms visualised by indirect immunofluorescence. Today, the primary sequence elements that direct murine CMP-NeuAc synthetase into the cell nucleus are known, however, information regarding the physiological relevance of the nuclear destination is still not available. With this article, we provide a detailed review on earlier and recent findings that identified and confirmed the unusual subcellular localisation of the CMP-NeuAc synthetase. In addition, we take the advantage to discuss most recent developments towards understanding structure--function relations of this enzyme.

Amino Acid Sequence↗
PubMed2000

Visualisation of three-dimensional microcracks in compact bone.

Microdamage in bone contributes to the loss of bone quality in osteoporosis and is thought to play a major role in both fragility and stress fractures (Schaffler et al. 1995). In this study, in vivo microcracks in human ribs were bulk-stained in basic fuchsin and viewed in longitudinal section and in 3 dimensions using 2 different computer-based methods of reconstruction: (1) serial sectioning of methylmethacrylate embedded sections using a sledge macrotome and identification of microcracks using UV epifluorescence followed by computerised reconstruction of microcracks using software and (2) laser scanning confocal microscopy of thick sections followed by reconstruction of microcracks into a 3-D image. The size and shape of microcracks were found to be similar using both techniques. Both techniques of reconstruction showed microcracks to be approximately elliptical in shape. From the serial sectioning reconstructions (n = 9), microcracks were found to have a mean length of 404 +/- 145 microm (mean +/- S.D.) (in the longitudinal direction) and mean width of 97 +/- 38 microm (in the transverse direction). Using epifluorescence microscopy, 92 microcracks were identified; mean microcrack length was 349 +/- 100 microm in the longitudinal direction. This was consistent with other results (Burr & Martin, 1993) and with the theoretical prediction of an elliptical crack shape with aspect ratio (longitudinal: transverse) of 5:1 deduced from analysis of random 2-D sections (Taylor & Lee, 1998). The results obtained provide new data on the nature of microcracks in bone and the method has the potential to become a useful tool in the calculation of stress intensity values which indicate the probability of an individual microcrack propagating to cause a stress or fragility fracture.

Aged↗
PubMed1994

Visualisation by electron microscopy of the unique part of the cytoplasmic domain of a desmoglein, a cadherin-like protein of the desmosome type of cell junction.

Part of the cytoplasmic domain of a human desmoglein, Dsg1, a cadherin-like protein found in desmosomes of epithelial cells, has been visualised by electron microscopy. The cloned fragment contains five repeats of a 29 +/- 4 residue sequence unique to desmogleins, followed by a glycine-rich region. In rotary shadowed preparations the molecule consists of a globular head attached to a thin tail, the latter perhaps corresponding to the glycine-rich region. This portion of the molecule is thought to span the width of the inner dense plaque. The structure and dimensions concur well to the configuration deduced from the protein sequence.

Amino Acid Sequence↗
PubMed2004

On-line variable live-adjusted displays with internal and external risk-adjusted mortalities. A valuable method for benchmarking and early detection of unfavourable trends in cardiac surgery.

OBJECTIVE: Benchmarking and early detection of unfavourable trends. METHODS: We implemented a dedicated project-orientated data warehouse, which continuously supplies data for on-line computing of the variable live-adjusted displays (VLADs). To calculate the expected cumulative mortality, we used the multi-variate logistic regression model of the EuroSCORE model. In addition to the external EuroSCORE standard, we calculated a centre-specific risk score for internal standards by analysing the data of 9135 patients, which enables both internal and external comparisons. The VLADs are embedded into the multi-purpose web-based information portal, so that the physicians can investigate several types of VLADs interactively: performance of different types of surgery and individual surgeons for different time intervals. We investigated clinically important events such as modification of operative techniques and personnel changes of the team by the VLADs. RESULTS: We found transient declines in the performance curves during major changes in patient management, indicating that systemic--rather than accidental or patient related factors--were involved in the mortality risk. The internal standard line represents these clusters more clearly than the external line. We evaluated examples of how periods of increased risk could be monitored by the VLAD curves: (1) the introduction of OPCAB surgery; (2) training of surgeons; (3) staff changes and staff-related management. CONCLUSIONS: On-line VLADs based on a day-to-day updated database, displaying both internal and external standards, are a helpful visualisation tool for earlier detection of unfavourable trends. They enable the surgeon teams and clinical management to take countermeasures at an early stage.

Benchmarking↗
PubMed1996

Chronic sputum production: correlations between clinical features and findings on high resolution computed tomographic scanning of the chest.

BACKGROUND: There are few published data on the correlation between the clinical findings in subjects with chronic sputum production and the appearances on high resolution computed tomographic (HRCT) scans of the chest. METHODS: HRCT scanning of the chest was performed on 40 subjects with chronic sputum production. Three readers independently reported the scans for the presence or absence of bronchiectasis and the extent of bronchiectasis on the basis of the percentage of involved bronchi in each lobe. Relationships were sought between these findings and the clinical history, physical examination, and laboratory investigations. RESULTS: HRCT scanning showed that 27 subjects had bronchiectasis. Of the clinical features only the continual production of purulent sputum and childhood pertussis were associated with bronchiectasis. There was a positive correlation between the extent of bronchiectasis and dyspnoea, and a negative correlation with forced expiratory volume in one second but not with forced vital capacity. CONCLUSIONS: These results indicate that, in subjects with chronic sputum production, only a few clinical features show any correlation with the presence or extent of bronchiectasis as visualised on HRCT scans.

Adult↗
PubMed2000

Technetium-99m RP527, a GRP analogue for visualisation of GRP receptor-expressing malignancies: a feasibility study.

Gastrin-releasing peptide (GRP) receptor scintigraphy could allow prediction of response to GRP receptor-targeted treatment options, early non-invasive diagnosis and in vivo prognostic stratification of GRP receptor-positive tumours. This study reports on the imaging characteristics and efficacy for tumour detection of technetium-99m RP527, a 99mTc chelated targeting peptide derived from bombesin, which binds GRP receptors with high affinity. Ten patients (four men and six women, mean age 56.4 years) either suffering from metastasised prostate (n, number of patients = 4) or breast carcinoma (n=1) or presenting with a clinical diagnosis highly suggestive for breast carcinoma (n=5) were included in the study. In the latter five patients, 99mTc-RP527 scintigraphy was performed prior to diagnostic, e.g. biopsy, and staging examinations. Final diagnosis in these patients was breast carcinoma in all five. In all patients, whole-body planar scans and tomographic images were acquired 1 h and 5-6 h post injection of 555 MBq 99mTc-RP527 and tumour to normal tissue (T/N) ratios determined. 99mTc-RP527 showed specific uptake in four of six breast and one of four prostate carcinomas. T/N ratios derived from planar and tomographic images increased significantly (P<0.01) from 1.65 (SD 1.53) and 3.35 (SD 3.04) to 2.58 (SD 1.26) and 7.23 (SD 8.46), respectively. T/N ratios derived from tomographic images were consistently higher (P<0.01). The data presented suggest that 99mTc-RP527 results in specific tumour localisation and exhibits good imaging characteristics with a good T/N ratio that may be further enhanced by single-photon emission tomography.

Adult↗
PubMed2004

Inactivated enzymes as probes of the structure of arabinoxylans as observed by atomic force microscopy.

The complex structures of water-soluble wheat arabinoxylans have been mapped along individual molecules, and within populations, using the visualisation of the binding of inactivated enzymes by atomic force microscopy (AFM). It was demonstrated that site-directed mutagenesis (SDM) can be used to produce inactive enzymes as structural probes. For the SDM mutants AFM has been used to compare the binding of different xylanases to arabinoxylans. Xylanase mutant E386A, derived from the Xyn11A enzyme (Neocallimastrix patriciarium), was shown to bind randomly along arabinoxylan molecules. The xylanase binding was also monitored following Aspergillus niger arabinofuranosidase pre-treatment of samples. It was demonstrated that removal of arabinose side chains significantly altered the binding pattern of the inactivated enzyme. Xylanase mutant E246A, derived from the Xyn10A enzyme (Cellvibrio japonicus), was found to show deviations from random binding to the arabinoxylan chains. It is believed that this is due to the effect of a small residual catalytic activity of the enzyme that alters the binding pattern of the probe. Control procedures were developed and assessed to establish that the interactions between the modified xylanases and the arabinoxylans were specific interactions. The experimental data demonstrates the potential for using inactivated enzymes and AFM to probe the structural heterogeneity of individual polysaccharide molecules.

Carbohydrate Sequence↗
PubMedDate not supplied

Breast cancer imaging with radiolabelled peptide from complementarity-determining region of antitumour antibody.

Specific tumour imaging with radiolabelled monoclonal antibodies has been extensively investigated. Although some success has been reported, there are many limitations due to the slow kinetics, poor extravasation, catabolism by the reticuloendothelial system, and non-specific uptake of macromolecules such as antibodies. We have tried to overcome some of the problems associated with monoclonal antibodies while retaining their specificity by using an antibody-derived synthetic peptide. A synthetic pentadecapeptide (alpha M2) derived from the third heavy-chain complementarity-determining region (CDR-3H) of a tumour-associated monoclonal antibody was produced and shown to retain its specificity against the pan-carcinoma cell-surface antigen, polymorphic epithelial mucin, detected by the parent antibody. The peptide was radiolabelled with technetium-99m and injected intravenously to image malignant lesions in 26 women with primary, recurrent, or metastatic breast cancer. Visualisation of breast tumours and their metastases was obtained shortly after administration of alpha M2, and was optimum by 3 h. Overall, 57 (77%) of 74 sites were visualised. Successful imaging was achieved in 14 of 15 primary tumour sites and all of eight local recurrences. Five of six metastases in the opposite breast, eight of 15 metastatic axillary lymph nodes, and all of six metastatic supraclavicular lymph nodes were imaged. Metastatic sites in the lungs, mediastinum, chest wall, and liver were poorly visualised because of background cardiac blood pool. alpha M2 detected small lesions ( < 2 cm) as efficiently as larger ones. The peptide was rapidly (3 h) cleared from the circulation. No acute or chronic adverse reactions due to the alpha M2 were observed. Specific tumour targeting with the radiolabelled anticancer peptide alpha M2 offers new opportunities for breast cancer imaging and possibly therapy.

Adult↗
PubMed2004

Phase behaviour and morphology of binary mixtures of DPPC with stearonitrile, stearic acid, and octadecanol at the air-water interface.

The behaviour of dipalmitoylphosphatidylcholine (DPPC), mixed with stearonitrile (SN), was investigated at the air-water interface by surface pressure-area (pi-A) measurements and by direct visualisation of monolayers by Brewster angle microscopy (BAM). The pi-A-X diagram of system DPPC/SN was compared with the corresponding diagrams of systems DPPC/stearic acid (SA) and DPPC/octadecanol (OD) at 20 degrees C. Monolayers of the three systems reach the closest packing of alkyl chains in the 0.4-0.6 range of XDPPC. Thermodynamic analysis indicates miscibility in the three binary systems with negative deviations from the ideal behaviour. Morphological features of system DPPC/SN change significantly with XDPPC and temperature in the range 10-30 degrees C. At 10 and 20 degrees C mixed monolayers form condensed states from low pi all over the composition range. At 30 degrees C, the liquid-expanded (LE)--liquid-condensed (LC) phase transition occurs at increasing pi with XDPPC. The shape and size of condensed domains change with XDPPC and pi. Contrarily to the behaviour of pure components, mixed monolayers of DPPC/SN exhibit orientational order in the 0.2-0.6 mol fraction range of DPPC. BAM observation confirmed the partial miscibility indicated by GE data in a limited range of compositions at 30 degrees C.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1997

Polymorphic curvature of satellite DNA in three subspecies of the beetle Pimelia sparsa.

The curvature of the monomeric repeats of satellite DNAs from three subspecies of the beetle Pimelia sparsa (Coleoptera, Tenebrionidae) has been analysed. Evidence of curvature was inferred from their retarded migration in native polyacrylamide gels, which was confirmed by direct electron microscopy visualisation. Sequence-comparison analysis, which included sequence alignments and modelling studies, was used to reveal the patterns of local bending and curvature. The effects of the minor-groove-binding drugs distamycin and berenil on the curvature were assayed, and analysed with respect to their (A+T)-rich sequence preferences. Since our study deals with satellite DNAs from closely related organisms (three subspecies), we correlated the differences in sequence, and also the high similarity conserved in the (A+T)-rich regions, with the changes in the patterns of curvature.

Animals↗
PubMed2003

Prognosis in node-negative primary breast cancer: a neural network analysis of risk profiles using routinely assessed factors.

BACKGROUND: The present study investigated complex time-dependent effects of routinely assessed factors on the risk of breast cancer recurrence over follow-up time, with a partial logistic artificial neural network (PLANN) model. PATIENTS AND METHODS: PLANN was applied to data from 1793 patients with node-negative breast cancer, not submitted to any adjuvant treatment and with a minimal potential follow-up of 10 years. RESULTS: The shape of the hazard function changed according to histology, which showed a time-dependent effect, partly modulated by estrogen receptors (ERs). Age and progesterone receptors (PgR) showed protective effects; the latter was more evident for short follow-up and high ER values. Tumour size and ER content showed time-dependent unfavourable effects at early and long follow-up times, respectively. Predicted values of disease recurrence probability at 2 years of follow-up showed that low steroid-receptor content, young age and large tumour size were associated with the highest risk of relapse. Although the oldest patients with high ER content seem to be those most protected overall, high risk predictions tend to spread also to higher steroid-receptor contents, intermediate ages and small tumour size, with an increase in follow-up time. CONCLUSION: PLANN with suitable visualisation techniques provided thorough insights into the dynamics of breast cancer recurrence for improving individual risk staging of node-negative breast cancer patients.

Breast Neoplasms↗
PubMed2004

Radiolabeled tracers for imaging of tumor angiogenesis and evaluation of anti-angiogenic therapies.

A variety of therapeutic strategies in oncology are focused on the inhibition of tumor-induced angiogenesis. Thus, there is a keen interest in methods which allow non-invasive monitoring of molecular targets involved in angiogenesis which would support information for planning and controlling corresponding therapies. Moreover, such techniques would provide an insight into the formation of new sprouting blood vessels, the involved processes and regulatory mechanisms in patients. At the moment, development of radiotracer based techniques is mainly concentrated on three different targets which include peptidic and non-peptidic alpha v beta 3-integrin binding antagonists, matrix metalloproteinase inhibitors and single chain anti-fibronectin antibody fragments. Development of radiolabeled MMP inhibitors is based on either the decapeptide Cys-Thr-Thr-His-Trp-Gly-Phe-Thr-Leu-Cys resulting from a phage display library or small molecular weight compounds. The in vitro data for these tracers are very promising. However, more detailed in vivo data are necessary to evaluate the potency of MMP-inhibitors for in-vivo imaging. The radiolabelled anti-ED-B single chain antibody fragment scFv L-19 shows selective accumulation in the tumor vasculature in a murine tumour model. In a first patient study a selective localisation of the (123)I-labeled tracer in lesions of different tumours was found. On the basis of the lead structure cyclo(-Arg-Gly-Asp-dPhe-Val) a variety of different radiolabeled RGD-peptides has been developed for the non-invasive determination of the alpha v beta 3 expression. These developments include peptides labeled with minimum structural alteration, peptide carbohydrate conjugates, peptidomimetics based on the RGD-structure as well as heterodimeric, homodimeric and homotetrameric ligand systems. Many of the tracers show high alpha v beta 3-affinity and selectivity in vitro and receptor selective tumour accumulation with high image contrast in different murine tumour models. Further studies have to demonstrate that this approach can be translated to clinical settings allowing visualisation of alpha v beta 3-positive tumours and alpha v beta 3 expression during tumour-induced angiogenesis in patients.

Angiogenesis Inhibitors↗
PubMed2003

Radiotracer-based strategies to image angiogenesis.

Tumour-induced angiogenesis plays an important role in tumour progression. Great efforts are made to develop therapeutic strategies to interfere with this process resulting in the starvation of the tumour. However, strategies to monitor conventional therapies seems to be inappropriate to control these approaches. Thus, there is a keen interest in developing methods supplying information about the corresponding therapeutical effects. Several radiotracer-based approaches focused on different targets in the angiogenic process are currently investigated. One class of tracers is based on matrix metalloproteinases inhibitors. These compounds show promising results in in vitro assays. However, initial data from in vivo studies using murine tumour models could not confirm successful non-invasive monitoring of MMP activity yet. Another strategy uses a radiolabelled single chain fragment against the ED-B domain of fibronectin, an extracellular matrix protein. Promising results demonstrated selective accumulation of the tracer in the tumour vasculature of a murine tumour model. Most of the studies are concentrated on the development of radiolabelled antagonists of the integrin alpha(v)beta(3). This heterodimeric transmembrane glycoprotein is involved in the migration of activated endothelial cells during formation of new vessels. Different compounds have been labelled with (18F), (111)In, (99m)Tc, (90)Y and several iodine isotopes. In in vitro assays most of them revealed high alpha(v)beta(3) affinity and selectivity. Moreover, in different murine tumour models successful non-invasive determination of alpha(v)beta(3) expression has been shown. Some of these approaches indicate that tumour-induced angiogenesis can be monitored in animal studies. Nevertheless, translation of these approaches into clinical settings allowing visualisation of tumour-induced angiogenesis in patients needs still to be demonstrated.

Animals↗