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Multi-dimensional visualisation of laboratory findings and functional test results for analysing the clinical course of disease in medicine.

The illustration of a patient's history by a graphical primitive is discussed. Illustration technology is presented which simultaneously represents quantitative examination findings (e.g., laboratory values) and qualitative findings (e.g., from function diagnostics) by a single geometrical figure. Depending on the medical results, this figure takes on characteristic forms which can be identified as patterns typical for a specific disease. The procedure developed is integrated in a user interface which is implemented in the form of a computerized medical record for use on a pentop computer. This portable computer assists the physician during ward rounds, supplies additional, intelligence-based information, serves quality control, and streamlines working procedures making them more efficient.

Clinical Laboratory Information Systems↗

A functional bipartite nuclear localisation signal in the cytokine interleukin-5.

Interleukin (IL)-5 is central in regulating eosinophilia in allergic disease and parasitic infections. We have identified a bipartite nuclear localisation signal (NLS) within amino acids 95-111 of human IL-5 (hIL-5), also present in mouse IL-5 (mIL-5). hIL-5 and mIL-5 were labelled fluorescently, and nuclear uptake subsequent to membrane binding and internalisation by intact receptor expressing cells visualised and quantified using confocal laser scanning microscopy. hIL-5 and mIL-5 were shown to be transported to the nucleus in in vivo and in vitro nuclear protein import assays. The hIL-5 NLS was able to target a heterologous protein to the nucleus both in vivo and in vitro. Mutations within the proximal arm of the NLS abrogated nuclear targeting activity, confirming its bipartite nature. The results imply a nuclear signalling role for IL-5 additional to pathways linked to the membrane receptor system.

Amino Acid Sequence↗

GENOLYTE: a PC-based computer program for visualisation of genomic DNA sequences.

This paper describes the program "GENOLYTE" which has been developed for the visualisation and identification of the global and local patterns of long genomic DNA sequences quickly in a few minutes, with the aid of a microcomputer. Apart from global and local identification, GENOLYTE comparatively depicts the similarity of two or more than two genomic DNA sequences. The utility of the program has been demonstrated by analysing the complete mitochondrial DNA sequences (taken from EMBL databank) as an example and the results are discussed from therein. The program written in TURBO C++ has the following minimum requirements: (i) PC 386 AT, (ii) SVGA colour monitor, (iii) the [BGI] directory of TURBO C++ version 3.0.

Animals↗

99mTc-Demotate 1: first data in tumour patients-results of a pilot/phase I study.

Somatostatin receptor (SSTR) scintigraphy with indium-111 DTPA-octreotide has become a routine diagnostic procedure in oncology. However, it suffers some drawbacks concerning the limited availability, suboptimal imaging properties and elevated radiation burden of (111)In. We have recently been involved in the development of a new tetraamine-functionalised [Tyr(3)]octreotate derivative (Demotate 1) that can be easily labelled with technetium-99m at high specific activities. (99m)Tc-Demotate 1 showed promising properties in preclinical studies. In this study we report on the first experience with (99m)Tc-Demotate 1 in patients. Six patients (mean age 56 years) with carcinoid tumours ( n=2) or endocrine pancreatic tumours ( n=4) with previously positive SSTR scintigraphy were enrolled in the study. Patients were injected with 500-600 MBq (99m)Tc-Demotate 1. Clinical and laboratory parameters were controlled up to 3 months p.i. Blood samples were taken at various time points up to 24 h p.i., and urine was collected up to 24 h. Whole-body images were acquired at 15-30 min, 1-2 h, 4 h and 24 h p.i. with additional single-photon emission tomography imaging at 1-4 h. Blood excretion was very rapid, with <2%ID in plasma after 1 h, and urinary excretion <20% ID after 6 h. Two patients complained of mild gastrointestinal problems and paraesthesia, but no other adverse reactions were observed. SSTR-positive tumours were rapidly visualised as early as 15 min p.i., with maximum tumour uptake (up to 19% ID) and tumour/organ ratios as early as 1 h p.i. Organs of predominant physiological uptake were the spleen and the kidneys, with no intestinal excretion detectable up to 24 h. (99m)Tc-Demotate 1 detected 11 lesions while In-Oct detected ten; differences in uptake behaviour were observed in three patients. This study shows for the first time that peptides derivatised with a tetraamine ligand for labelling with (99m)Tc show suitable properties for receptor imaging in patients. (99m)Tc-Demotate 1 is a promising agent for the visualisation of SSTR-positive lesions in patients, allowing rapid imaging as early as 1 h p.i.; some differences are observed in pharmacokinetic behaviour compared with (111)In-DTPA-octreotide.

Aged↗

Baculovirus-mediated high level expression of a human thiopurine methyl transferase.

We have expressed the human thiopurine methyltransferase cDNA in a baculovirus vector in Sf21 (Spodoptera frugiperda) cells. This system expresses the enzyme at levels such that the thiopurine methyltransferase enzyme may be readily visualised by Coomassie blue stained sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The expressed enzyme catalysed the methylation of 6-mercaptopurine with an apparent Km of 892 microM, similar to that observed in human liver cytosol ie. 657 microM however, the Vmax was 13,500 pmole/mg/min, which is approximately 400 times higher than the Vmax observed in human liver cytosol ie. 33 pmole/mg/min. The thiopurine methyltransferase inhibitors 6-thioxanthine, p-methoxybenzoic acid and 3,5-dimethoxy benzoic acid were found to be potent inhibitors of the expressed enzyme.

Animals↗

[Localisation of parathyroid glands using planar (99m)Tc-sestamibi scintigraphy. Comparison between subtraction- and dual-phase technique].

AIM: In the context of presurgical localisation of parathyroid adenomas in primary hyper-parathyreoidism (pHPT) using (99m)Tc-sestamibi scintigraphy, subtraction- and dualphase technique are compared with each other and with the surgical findings. PATIENTS, METHODS: Prospectively, 126 patients with pHPT were investigated presurgically. For visualisation of parathyroid adenomas, an image of the thyroid ((99m)Tc-pertechnetat) was subtracted from a perfusion image ((99m)Tc-sestamibi) and 2 h p. i. another image was acquired for identification of retention of activity. Considering both techniques the clinical findings were reported promptly. Retrospectively, the evaluations were presented separately to four experienced raters. RESULTS: In clinical routine for 109 patients correct findings were reported presurgically (87%). From 129 resected parathyroid adenomas 118 were localised correctly (sensitivity 91%, positive predictive value 94%). Concerning the retrospective analysis, in 75% of the cases both techniques provided the correct site, in 14% only the dual-phase technique and in 7% only the subtraction-technique was correct. With the help of the dual-phase technique significantly more investigations were correctly rated than with the help of the subtraction technique (88.7 +/- 3.2% vs. 81.6 +/- 1.2%, p < 0.01, two-sided t-test). CONCLUSION: The presurgical scintigraphic localisation of hyperactive parathyroid glands in pHPT assists minimal invasive surgery serving a high rate of correct findings. According to our data the dual-phase technique seems to be more sensitive than the subtraction technique. In some cases, however, the correct site may only be found using the subtraction technique. For an optimal surgical strategy we suggest the combination of both techniques.

Humans↗

Three-dimensional MR imaging in the assessment of physeal growth arrest.

The purpose of this study is to describe an imaging method for identifying and characterising physeal growth arrest following physeal plate aggression. The authors describe the use of three-dimensional MRI performed with fat-suppressed three-dimensional spoiled gradient-recalled echo sequences followed by manual image reconstruction to create a 3D model of the physeal plate. This retrospective series reports the analysis of 33 bony physeal bridges in 28 children (mean age 10.5 years) with the use of fat-suppressed three-dimensional spoiled gradient-recalled echo imaging and 3D reconstructions from the source images. 3D reconstructions were obtained after the outlining was done manually on each source image. Files of all patients were reviewed for clinical data at the time of MRI, type of injury, age at MRI and bone bridge characteristics on reconstructions. Twenty-one (63%) of the 33 bridges were post-traumatic and were mostly situated in the lower extremities (19/21). The distal tibia was involved in 66% (14/21) of the cases. Bridges due to causes other than trauma were located in the lower extremities in 10/12 cases, and the distal femur represented 60% of these cases. Of the 28 patients, five presented with two bridges involving two different growth plates making a total of 33 physeal bone bars. The location and shape of each bridge was accurately identified in each patient, and in post-traumatic cases, 89% of bone bars were of Ogden type III (central) or I (peripheral). Reconstructions were obtained in 15 min and are easy to interpret. Volumes of the physeal bone bridge(s) and of the remaining normal physis were calculated. The bone bridging represented less than 1% to 47% of the total physeal plate volume. The precise shape and location of the bridge can be visualised on the 3D reconstructions. This information is useful in the surgical management of these deformities; as for the eight patients who underwent bone bar resection, an excellent correspondence was found by the treating surgeon between the MRI 3D model and the per-operative findings. Accurate 3D mapping obtained after manual reconstruction can also visualise very small physeal plates and bridges such as in cases of finger physeal disorders. MR imaging with fat-suppressed three-dimensional spoiled gradient-recalled echo sequences can be used to identify patterns of physeal growth arrest. 3D reconstructions can be obtained from the manual outlining of source images to provide an accurate representation of the bony bridge that can be a guide during surgical management.

Adolescent↗

Visualising the dissociation of sequence selective ligands from individual binding sites on DNA.

We have used a modification of the footprinting technique to measure the dissociation of mithramycin, echinomycin and nogalamycin from their binding sites in a natural DNA fragment. Complexes with radiolabelled DNA were dissociated by addition of unlabelled DNA. Samples were removed at various times and subjected to DNase I digestion, and the rate of dissociation from each site was estimated from the time-dependent disappearance of the footprints. For echinomycin the slowest rate of dissociation is from ACGT, while the slowest site for mithramycin contains four contiguous guanines. The dissociation of nogalamycin is extremely slow, even from its weaker sites; the slowest rate was from ACGTA, which took longer than 4 h, even at 37 degrees C.

Base Sequence↗

The role of two-dimensional echocardiography in the detection of potentially embolic intracardiac masses in patients with cerebral ischaemia.

The M-mode and two dimensional echocardiographic data of 62 consecutive cardiac patients referred from neurology centres were analysed retrospectively to establish the use of these techniques in detecting underlying cardiac pathology. All patients had presented initially to a neurologist with transient or permanent focal cerebral or retinal ischaemia, and had been referred for cardiac assessment after neurological investigations failed to establish the underlying cause of the neurological event. Patients were divided into two groups. In 30 patients the referring neurologist had found no evidence of cardiac disease (Group I); in the other 32 patients either heart disease or an arrhythmia had been diagnosed prior to cardiac referral (Group II). One of the patients in Group I had echocardiographic evidence of mitral valve prolapse not detected by the neurologist prior to referral; no cardiac pathology was recognised in the other 29 patients in this group. In seven of the 32 (22%) patients from Group II, a cardiac mass presumed responsible for the neurological manifestations was demonstrated echocardiographically, and in six of these histological confirmation was obtained following surgery or at necropsy. Two dimensional echocardiography was the only investigation which visualised the intracardiac pathology in four patients. In the remaining three patients, valve vegetations (two cases) and an atrial tumour (one case) were demonstrated by both echocardiographic methods. In patients with either clinical evidence of cardiac disease or an arrhythmia who have experienced one or more episodes of cerebral or retinal ischaemia, the presence of an intracardiac mass is not uncommon. Two dimensional echocardiography was the method of choice for detecting cardiac thrombus but the use of both methods of ultrasound should be considered as complementary techniques in the investigation of these cases. Routine echocardiography is unlikely to be of value in screening patients who have had a cerebrovascular event and who do not have clinical evidence of heart disease or an arrhythmia.

Adolescent↗

[Evaluation of diastolic function of in patients addicted to recreational cocaine].

BACKGROUND AND AIMS: The recreational use of cocaine is associated with cardiovascular pathologies, such as ischemic cardiopathy, myocarditis and cardiomyopathies, owing to the increased catecholamine stimulus, a propensity to coronary spasm, increased coagulative activity and inflammatory and degenerative phenomena of myocardiac cells. Early alterations of the diastolic phase may be visualised by evaluating the diastolic Doppler pattern of left ventricular filling. METHODS: For this purpose the authors compared blood pressure, heart rate, heart mass, protodiastolic (E wave) and telediastolic (A wave) filling rate and their ratio (E/A) on the Dopper mitral diastolic profile in a group of 10 patients addicted to the recreational use of cocaine (mean age 33 +/- 7) with those of 10 normal subjects (mean age 34 +/- 2). RESULTS: Patients using cocaine presented mean systolic arterial blood pressures of 130 +/- 12 versus 127 +/- 8 in control subjects (p = ns); mean heart rate was statistically significant with 98 +/- 14 versus 76 +/- 12 in controls (p < 0.05). There were no differences in cardiac mass between the two groups. In cocaine addicts the speed of the E wave was significantly slower: 58.4 +/- 8.6 versus 73 +/- 7.4 cm/sec (p < 0.05), and the speed of the A wave was significantly higher: 70.5 +/- 10.5 versus 62.6 +/- 4.3 cm/sec (p < 0.05), when compared with normal controls subjects; the E/A ratio of cocaine addicts was lower (0.75 +/- 0.34) compared to normal subjects (1.07 +/- 0.7), (p < 0.05). CONCLUSIONS: These data show that patients addicted to the recreational use of cocaine show preclinical alterations of the left ventricular diastolic phase prior to the onset of clinically evident pathologies.

Adult↗

Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer.

Screening for specific biomarkers of early-stage detection of ovarian cancer is a major health priority due to the asymptomatic nature and poor survival characteristic of the disease. We utilised two-dimensional gel electrophoresis (2DE) to identify differentially expressed proteins in the serum of ovarian cancer patients that may be useful as biomarkers of this disease. In this study, 38 ovarian cancer patients at different pathological grades (grade 1 (n=6), grade 2 (n=8) and grade 3 (n=24)) were compared to a control group of eight healthy women. Serum samples were treated with a mixture of Affigel-Blue and protein A (5 : 1) for 1 h to remove high abundance protein (e.g. immunoglobulin and albumin) and were displayed using 11 cm, pH 4-7 isoelectric focusing strips for the first dimension and 10% acrylamide gel electrophoresis for the second dimension. Protein spots were visualised by SYPRO-Ruby staining, imaged by FX-imager and compared and analysed by PDQuest software. A total of 24 serum proteins were differentially expressed in grade 1 (P<0.05), 31 in grade 2 (P<0.05) and 25 in grade 3 (P<0.05) ovarian cancer patients. Six of the protein spots that were significantly upregulated in all groups of ovarian cancer patients were identified by nano-electrospray quadrupole quadrupole time-of-flight mass spectrometry (n-ESIQ(q)TOFMS) and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOFMS) as isoforms of haptoglobin-1 precursor (HAP1), a liver glycoprotein present in human serum. Further identification of the spots at different pathological grades was confirmed by Western blotting using monoclonal antibody against a haptoglobin epitope contained within HAP1. Immunohistochemical localisation of HAP1-like activity was present in malignant ovarian epithelium and stroma but strong immunostaining was present in blood vessels, areas with myxomatous stroma and vascular spaces. No tissue localisation of HAP1-like immunoreactivity was observed in normal ovarian surface epithelium. These data highlight the need to assess circulating concentration of HAP1 in the serum of ovarian cancer patients and evaluate its potential as a biomarker in the early diagnosis of ovarian cancer.

Biomarkers, Tumor↗

Systemic inflammation in COPD visualised by gene profiling in peripheral blood neutrophils.

BACKGROUND: The inflammatory process in chronic obstructive pulmonary disease (COPD) is characterised by the presence of neutrophils in the lung that are able to synthesise de novo several inflammatory mediators. The local chronic persistent inflammatory response is accompanied by systemic effects such as cytokine induced priming of peripheral leucocytes and muscle wasting. The preactivation or priming of peripheral blood neutrophils was used to gain more insight into the mechanisms of this systemic inflammatory response. METHODS: Gene arrays were performed on peripheral blood neutrophils obtained from healthy donors after stimulation in vitro with tumour necrosis factor (TNF)-alpha, granulocyte-macrophage colony stimulating factor (GM-CSF), or both. The expression of many inflammatory genes was regulated in these cells following stimulation. The expression of inflammatory genes in peripheral blood neutrophils in healthy subjects and those with COPD was measured by real time RT-PCR after stimulation with TNFalpha, GM-CSF, interleukin (IL)-8, fMLP, TNFalpha + GM-CSF, and lipopolysaccharide (LPS). RESULTS: The genes regulated in the gene array with TNFalpha/GM-CSF stimulated neutrophils included cytokines (such as IL-1beta), chemokines (such as IL-8), and adhesion molecules (such as ICAM-1). Disease severity as measured by forced expiratory volume in 1 second (FEV(1)) in COPD patients correlated with expression of several of these genes including IL-1beta (r = -0.540; p = 0.008), MIP-1beta (r = -0.583; p = 0.003), CD83 (r = -0.514; p = 0.012), IL-1 receptor 2 (r = -0.546; p = 0.007), and IL-1 receptor antagonist (r = -0.612; p = 0.002). CONCLUSIONS: These data are consistent with the hypothesis that progression of COPD is associated with the activation of neutrophils in the systemic compartment. De novo expression of inflammatory mediators by peripheral blood neutrophils suggests a pro-inflammatory role for these cells in the pathogenesis of COPD.

Cytokines↗

Immunocytochemical identification of oestrogen receptors in preoptic neurones containing calcitonin gene-related peptide in the male and female rat.

Using single- and double-labelling immunocytochemistry with antibodies specific for the oestrogen receptor and calcitonin gene-related peptide (CGRP), we have demonstrated oestrogen receptor immunoreactivity in the sexually dimorphic CGRP-immunoreactive (IR) population of the medial preoptic area (MPOA). In the short-term gonadectomised female approximately 80% of preoptic CGRP-IR neurones were immunoreactive for the oestrogen receptor. In short-term gonadectomised males, a small population of CGRP-IR cells was visualised in the MPOA only after colchicine treatment. Approximately 30% of CGRP-IR cells in the male were oestrogen receptor-IR, accounting for 2% of the total population of cells containing oestrogen receptors in this area. In the colchicine-treated female, it is estimated that 10-15% of preoptic oestrogen receptor-IR cells contain CGRP. These results indicate that CGRP is synthesised by preoptic neurones with oestrogen receptors. Furthermore, the identification of oestrogen receptors in the sexually dimorphic CGRP population suggests that these neurones may be directly linked with gonadal steroid-dependent, sex-specific functioning of the MPOA.

Animals↗

Organotypic brain slice cultures: an efficient and reliable method for neurotoxicological screening and mechanistic studies.

This paper reviews the current state of the use of organotypic brain slice cultures for neurotoxicological and neuropharmacological screening and mechanistic studies, as exemplified by excitotoxin application. At present, no in vitro systems have been approved by the regulatory authorities for neurotoxicity testing. For the evaluation of the slice culture method, organotypic hippocampal slice cultures were exposed to toxic doses of the excitotoxins, glutamate, N-methyl-D-aspartate (NMDA), kainic acid and 2-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA), and the glial toxin, DL-alpha-aminoadipic acid (DLAAA). Neuronal cell death was quantified by propidium iodide (PI) uptake, and visualised by Fluoro-Jade (FJ) staining. General cell death was monitored by lactate dehydrogenase (LDH) release into the culture medium. EC50 values for the different compounds, based on PI uptake after exposure for 48 hours in entire cultures, were: glutamate, 3.5 mM; DL-AAA, 2.3 mM; kainic acid, 13 microM; NMDA, 11 microM; and AMPA, 3.7 microM. In the slice cultures, the hippocampal subfields displayed the same differences in vulnerability as those observed in vivo. When subfield analysis was performed on the cultures, the CA1 subfield was most susceptible to glutamate, NMDA and AMPA, while CA3 was most susceptible to kainic acid. The amount of LDH release for DL-AAA was about four times that of L-glutamate, in accordance with the additional toxic effect on glial cells, which was also found by confocal microscopy to stain for FJ. In conclusion, it was found that organotypic brain slice culture, combined with standardised protocols and quantifiable markers, such as PI and FJ staining, is a relevant and feasible in vitro system for neurotoxicity testing. Considering the amount and quality of the available published data, it is recommended that the brain slice culture method could be subjected to pre-validation and formal validation for inclusion in a tiered in vitro neurotoxicity testing scheme to supplement and replace conventional animal tests.

Animal Testing Alternatives↗

Chitinolytic activities in Heligmosomoides polygyrus and their role in egg hatching.

The occurrence of chitin in the eggshell of Heligmosomoides polygyrus has been determined by histochemical and biochemical techniques. Approximately 5% of the egg dry weight was chitin. Staining with Calcofluor white showed the chitin in the eggshell to be more accessible to the stain after hatching or rupturing of the eggshell. Chitinolytic activity has been detected using fluorescent substrates in extracts of adult males (at low levels), females and eggs. Enzyme activity in situ, within the developing larvae, was visualised with the same substrates. It was localized in discrete granules about 1 micron in diameter which occurred as groups in areas of about 5 microns in diameter, in the posterior third of the larvae. The chitinolytic activity in the eggs increased with the age of the egg and was released into the medium when the eggs hatched. The chitinase activities were very sensitive to inhibition by allosamidin, a specific chitinase inhibitor, with an IC50 for the crude egg extract of 2.2 nM. However, treatment of eggs with 250 microM allosamidin resulted in a slowing but not cessation of egg hatching.

Acetylglucosamine↗

A study of the interaction of DAPI with DNA containing AT and non-AT sequences--molecular specificity of minor groove binding drugs.

The binding specificity of DAPI to DNA has been probed by analysing its interactions with DNA octamers consisting of different base sequences, which include adenine, guanine, 2-amino adenine and inosine, using molecular mechanics methods. Presence of AT and non-AT base pairs in the immediate vicinity of the binding site, containing AT and non-AT base pairs is also investigated. Results show that DAPI most prefers to bind to homopolymer of AT, and least to the duplex containing alternating GC bases. DAPI interacts with homopolymeric duplexes in two possible orientations related by 180 degrees with nearly same affinity. Affinity of DAPI towards DNA comprising the modified bases, inosine and 2-amino adenine, is in between these extremities. The binding affinity is reduced to some extent by the occurrence of non AT bases flanking the four base paired binding region. An interesting revelation is that one can visualise DAPI to form a hydrogen bond with O2 of cytosine indicating that the 2-amino group of purines does not per se sterically preclude DAPI from residing in the minor groove of B-DNA helix. On the other hand, repulsive nature of electrostatic interactions that prevail at the minor groove consequent to the presence of these sequences contribute decisively in preventing further diffusion of the drug. Thus, electrostatics, rather than hydrogen bonding to bases, seemingly play an important role in determining the specificity of interaction. The retention of drug binders in the minor groove and therefore recognition, is governed by the combined effect of these various forces.

Base Sequence↗

Permeability and route of entry for lipid-insoluble molecules across brain barriers in developing Monodelphis domestica.

1. We have studied the permeability of blood-brain barriers to small molecules such as [(14)C]sucrose, [(3)H]inulin, [(14)C]L-glucose and [(3)H]glycerol from early stages of development (postnatal day 6, P6) in South American opossums (Monodelphis domestica), using a litter-based method for estimating steady-state cerebrospinal fluid (CSF)/plasma and brain/plasma ratios of markers that were injected I.P. 2. Steady-state ratios for L-glucose, sucrose and inulin all showed progressive decreases during development. The rate of uptake of L-glucose into the brain and CSF, in short time course experiments (7-24 min) when age-related differences in CSF production can be considered negligible also decreased during development. These results indicate that there is a significant decrease in the permeability of brain barriers to small lipid-insoluble molecules during brain development. 3. The steady-state blood/CSF ratio for 3000 Da lysine-fixable biotin-dextran following I.P. injection was shown to be consistent with diffusion from blood to CSF. It was therefore used to visualise the route of penetration for small lipid-insoluble molecules across brain barriers at P0-30. The proportion of biotin-dextran-positive cells in the choroid plexuses declined in parallel with the age-related decline in permeability to the small-molecular-weight markers; the paracellular (tight junction) pathway for biotin-dextran appeared to be blocked, but biotin-dextran was easily detectable in the CSF. A transcellular route from blood to CSF was suggested by the finding that some choroid plexus epithelial cells contained biotin-dextran. 4. Biotin-dextran was also taken up by cerebral endothelial cells in the youngest brains studied (P0), but in contrast to the CSF, could not be detected in the brain extracellular space (i.e. a significant blood-brain barrier to small-sized lipid-insoluble compounds was already present). However, in immature brains (P0-13) biotin-dextran was taken up by some cells in the brain. These cells generally had contact with the CSF, suggesting that it is likely to have been the source of their biotin-dextran. Since the quantitative permeability data suggest that biotin-dextran behaves similarly to the radiolabelled markers used in this study, it is suggested that these markers in the more immature brains were also present intracellularly. Thus, brain/plasma ratios may be a misleading indicator of blood-brain barrier permeability in very immature animals. 5. The immunocytochemical staining for biotin-dextran in the CSF, in contrast to the lack of staining in the brain extracellular space, together with the quantitative permeability data showing that the radiolabelled markers penetrated more rapidly and to a much higher steady-state level in CSF than in the brain, suggests that lipid-insoluble molecules such as sucrose and inulin reach the immature brain predominantly via the CSF rather than directly across the very few blood vessels that are present at that time.

Algorithms↗

Endometrial destruction techniques for heavy menstrual bleeding.

BACKGROUND: Heavy menstrual bleeding (HMB) is a significant health problem in premenopausal women that can reduce quality of life and cause anaemia. First line therapy has traditionally been medical therapy but this is not always completely effective. Hysterectomy, often used after the failure of medical therapy, is 100% effective but is risky, costly and can cause complications. Endometrial ablation is less invasive, less costly and preserves the uterus. A large number of techniques have been developed to "ablate" (remove) the lining of the endometrium. The gold standard techniques (laser, transcervical resection of the endometrium and rollerball) require visualisation of the uterus with a hysteroscope and, although safe, require skilled surgeons. A number of newer techniques have recently been developed, most of which can be performed blind and are less time consuming. Many of these techniques are still under development, refinement and investigation. OBJECTIVES: To compare the efficacy, safety and acceptability of methods used to destroy the endometrium to reduce HMB in premenopausal women. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (issue 4, 2001), Medline (1966 to September 2001), EmBase (1980 to August 2001), Current Contents (1993 to week 38, 2001), Biological Abstracts (1980 to June 2001), Psyclit (1967 to August 2001) and Cinahl (1982 to July 2001). We also searched the specialised register of the Cochrane Menstrual Disorders and Subfertility Group (August 2001). We also searched reference lists of articles and contacted pharmaceutical companies and experts in the field. SELECTION CRITERIA: Randomised controlled trials comparing endometrial ablation techniques in women with a complaint of heavy menstrual bleeding without uterine pathology. The outcomes included reduction of heavy menstrual bleeding, improvement in quality of life, operative outcomes, satisfaction with outcome, complications and need for further surgery. DATA COLLECTION AND ANALYSIS: The two reviewers independently selected trials for inclusion, assessed trials for quality and extracted data. Attempts were made to contact authors for clarification of data in some trials. Adverse events were only assessed if they were separately measured in the included trials. MAIN RESULTS: In comparing hysteroscopic techniques, the vaporising electrode procedure was less difficult to perform (OR=0.25, 95%CI 0.1, 0.7) and had less fluid deficit (WMD=-258mls, 95% CI -342.1, -174.0) than TCRE. The odds of fluid overload and equipment failure were higher ((OR=5.2, 95% CI 1.5, 18.4) and (OR=6.0, 95% CI 1.7, 20.9) respectively) for those women having laser as compared to TCRE (transcervical resection of the endometriuim). In comparing traditional hysteroscopic endometrial ablation with the newer 2nd generation techniques overall, the newer techniques took less time to perform (WMD=-11mins, 95% CI -18.6, -2.6) and were more likely to be performed under local anaesthesia (OR=7.6, 95% CI 1.1, 52.7) but had a greater chance of equipment failure (OR=4.1, 95% CI 1.1, 15.0). The reduction in heavy bleeding did not differ significantly between any of the groups. REVIEWER'S CONCLUSIONS: Endometrial ablation techniques continue to play an important role in the management of HMB. The rapid development of a number of new methods of endometrial destruction has made systematic comparisons between methods and with the "gold standard" of TCRE difficult. Most of the newer techniques are performed blind and are technically easier than hysteroscopy-based methods. Overall, the existing evidence suggests success rates and complication profile of newer techniques of ablation compares favourably with TCRE, although technical difficulties with new equipment need to be ironed out.

Adult↗