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[Should adrenalectomy be performed systematically as part of radical nephrectomy for renal cancer? Review of 119 cases].

Systematic histological examination of 119 operative specimens of radical nephrectomies performed for renal cell carcinoma revealed adrenal invasion in 6 cases (5.04%). Preoperative computed tomography visualised the adrenal lesions with a sensitivity of 100%. Adrenal involvement was due to either contiguous invasion (3 cases) or metastatic spread (3 cases). Other visceral or lymph node metastases were detected in every case. The mean survival of patients with an adrenal lesion was 16.5 months. These data and those reported in the literature suggest the value of systematic adrenalectomy as part of radical nephrectomy.

Adrenalectomy

Improved subtraction technique in intravenous digital left ventriculography: comparison with radionuclide studies.

Adequate processing of left ventricular angiograms depends on the visualisation of all segments of the ventricular wall. At the same time, subtraction of different images can enhance different heart segments but commercially available methods do not allow simultaneous viewing of several images masked by different processes. Using our software, for each studied frame, a four quadrant display permits the simultaneous visualisation of a mask mode image, a diastolic-systolic difference image, an image obtained by subtraction of a frame at the same cycle time and a composite mask subtracted image. The composite mask image is obtained by weighting three images according to videodensitometric measurements by reference to previously acquired data. This method facilitates contour delineation and computation of the ejection fraction by area-length method. Correlation with radionuclide estimates of left ventricular ejection fraction is higher (n = 60, r = 0.90, SEE = 8%) than using the classical mask mode display (n = 60, r = 0.82, SEE = 11%). In a subgroup of 30 patients the contrast medium was injected in an antecubital vein and the correlation coefficient remained satisfactory (n = 30, r = 0.89, SEE = 7%) when compared with the classical subtraction technique (n = 30, r = 0.70, SEE = 12%). We therefore conclude that the composite mask method gives comparatively similar values for left ventricular ejection fraction to those acquired by radionuclide angiography.

Angiography, Digital Subtraction

Electron microscope observations on the entry of avian infectious bronchitis virus into susceptible cells.

Infectious bronchitis virus was observed to enter cells of chicken chorioallantoic membrane by viropexis. There was no support for the suggestion that entry took place by fusion of viral and plasma membranes. The results of electron microscopy showed that virus attachment occurred both at 4 degrees and at 37 degrees C. Viropexis was not observed until the preparations were warmed. Similar results were obtained using chicken kidney cells. Quantitative data obtained from a plaque counting system employing chicken kidney cells indicated that attachment was the same at both temperatures and that some virus particles were taken up at 4 degrees C. Virus uptake was triggered by attachment of the virus to the cell membrane and the subsequent process of virus entry visualised by E. M. appeared to proceed without the involvement of lysosomal enzymes. No intracellular virus was located by electron microscopy in warmed preparations when virus was treated with specific antiserum, either before or after adsorption to the cells.

Adsorption

The in vitro slice preparation for combined morphological and electrophysiological studies of rat visual cortex.

The morphological condition of slices of rat visual cortex, maintained in vitro in an interface-type recording chamber, was assessed. In addition, neurones in these slices were impaled with glass micropipettes for intracellular recording and horseradish peroxidase (HRP) injection. After fixation and embedding, slices were examined by light and electron microscopy. Slices sectioned orthogonally to the original plane of cutting showed a vertical zonation of tissue preservation. The upper zone contained dense and flattened neuronal somata, although the neuropil appeared normal. The central zone was well preserved, with the appearance of most somata, dendrites, axons and synapses comparing favourably with perfusion-fixed material. The lower zone contained many abnormal, vacuolated somata. The morphology of HRP-injected neurones was assessed by light microscopy. Dendrites could be visualised in great detail and spines were clearly visible. Local axon arbors were well represented. There was good correlation between electrophysiological and morphological criteria for the assessment of the condition of the slice. We conclude that, provided the extent of degeneration within the slice is monitored and appreciated, slices of visual cortex can provide both electrophysiological and morphological data of high quality.

Animals

Laparoscopic removal of translocated intrauterine contraceptive devices.

Data is presented regarding 20 translocated intrauterine contraceptive devices (IUCD). Successful laparoscopic removal was made in 13 (65%) cases though all these devices were Copper 'T' 200 model which tend to produce dense adhesions. Only when the IUCD was suspected to be partially or fully in the gut lumen or thick adhesions were present, or the device was not visualised, was laparotomy performed. Hence it is advocated that laparoscopy should be performed as a routine in patients with suspected translocated IUCD, as it obviates the need for laparotomy and thus decreases the duration of the hospital stay as well as preventing morbidity associated with laparotomy.

Adult

Zone equalisation normalisation for improved alignment of epigenetic signal.

MOTIVATION: High-throughput genomic technologies have transformed our understanding of biological systems, yet direct comparison and visualisation of these complex datasets remains challenging. Existing normalisation methods often fail to align genomic signal across samples due to sensitivity to sequencing depth differences and localised high-signal artefacts, leading to inconsistent replicate behaviour and increased downstream variability. RESULTS: We introduce Zone Equalisation Normalisation (ZEN), a novel approach designed to improve cross-sample signal alignment of genomic data. ZEN rescales genomic signal based on variance estimated within biologically enriched regions, reducing the influence of extreme outliers while preserving underlying biological structure. Using a diverse collection of data and our new genome-wide benchmarking approach, we reveal that ZEN improves biological and technical replicate alignment across the majority of tested conditions and experimental platforms. We further show that this improved signal comparability is associated with fewer differential accessibility calls between technical replicates and a more conservative set of biological differences. Together, these results demonstrate that ZEN provides a complementary framework to improve the accuracy and reliability of genomic data analysis and that normalisation choice can affect downstream analyses and biological interpretation. AVAILABILITY AND IMPLEMENTATION: ZEN is available as an open-source Python package via conda and PyPI. Source code, documentation, tutorials, and code to reproduce the analyses are available at https://github.com/Genome-Function-Initiative-Oxford/Zone-Equalisation-Normalisation and Zenodo (https://doi.org/10.5281/zenodo.21067751).

Epigenesis, Genetic

Quantitative assessment of blood-brain barrier permeability in multiple sclerosis using 68-Ga-EDTA and positron emission tomography.

Fifteen patients with definite multiple sclerosis were examined with high volume delayed (HVD) CT scan and positron emission tomography (PET) using 68-Ga-EDTA as a tracer. The passage of 68-Ga-EDTA across the blood-brain barrier (BBB) was measured by using multiple graphical analysis. This method permits the simultaneous calculation of a blood to brain influx constant Ki (ml/g-1 min-1) and of the plasma volume Vp (ml/g-1). Focal areas of abnormal CT enhancement and pathological accumulation of 68-Ga-EDTA were visualised in four patients who were all examined during a clinical exacerbation of the disease. The mean Ki value measured in these areas was 12.5, SD 3.4 indicating a moderate but significant increase of BBB permeability compared with the value found in normal tissue (3.2, SD 0.9). No parallel increase in Vp values was found in these pathological areas. Quantitative data obtained with PET seems to provide further insight into the study of BBB function in multiple sclerosis.

Adolescent

Computer applications in orthopaedics.

With the rapid developments in microprocessors, the widespread availability of computers has brought about broad applications in the field of orthopaedics. The present technology enables large quantities of data to be logically processed in a very short span of time. This has led to the development of information management database systems where relevant medical information may be retrieved very quickly and effectively. The analytical power of the computer has also been utilised in expert systems to assist in clinical-decision making process. Computer graphics have revolutionised the visualisation of physical features of internal and external body parts, providing new and improved modalities of diagnosis. In some centres, surgical planning and rehearsals are already being carried out at the computer terminal with the use of animation and computer graphics. Computer technology has also played an active role in the field of prosthetics and rehabilitation. Intelligent robotic systems and microprocessor with functional neuromuscular stimulation have been applied to benefit, and in some cases restore some motor functions to the physically disabled. With more collaboration between engineers, scientists and the medical community, several prototypes of computer-controlled prostheses and prosthesis designed and manufactured by Computer-Aided Design/Computer-Aided Manufacturing (CAD/CAM) technology are available today to assist the amputees in their daily living and ambulatory activities.

Computer Graphics

The relation between induced reciprocal translocations and cell killing of mouse spermatogonial stem cells after combined treatments with hydroxyurea and X-rays.

The induction of reciprocal translocations in mouse spermatogonial stem cells, visualised in dividing primary spermatocytes, was studied after combined treatments with hydroxyurea (250 and 500 mg/kg) and X-rays (6, 8 and 9 Gy). The time intervals between the 2 treatments were 16 h (leading to extremely high cell killing) and 48 h (giving rise to less killing than irradiation alone). Comparison of the observed frequencies of translocations with reported data on stem cell killing (de Ruiter-Bootsma and Davids, 1981) show that the ratio between the probabilities that a radiation-induced basic lesion kills a cell or produces a translocation, theoretically calculated by Leenhouts and Chadwick (1981) to be about 10, can indeed be confirmed experimentally.

Animals

[Possible applications of a method for 3D reconstruction of CT image data. OP-planning, finishing of individual alloplastic implants for replacement of cranial and maxillofacial bone structures].

A method for the three-dimensional reconstruction of CT-image data permits manufacturing of natural size anatomical models thus enabling preoperative measurements of relevant parameters (length, areas, and volumes). The methods described allow for new perspectives in preoperative planning of dental implants, particularly for the maxillary problem area. The use of 3D image data for manufacturing individual alloplastic implants simplifies the exact reconstruction of symmetrical bone structures, which up to now was impossible to achieve with conventional methods. Applications in the fields of orthodontic surgery and in neurological surgery are suggested. Refinement of available techniques is possible by means of additional segmentation of soft tissues, and by implementation of more sophisticated software for better visualisation and smoothly shaped surfaces. Future applications may even include aesthetic surgery.

Adult

Effects of isoprenaline and ouabain on cytosolic calcium and cell motion in single rat cardiomyocytes.

OBJECTIVE: The aim was to develop a novel system to measure changes in cytosolic free calcium concentration ([Ca2+]i) and cell motion simultaneously in a single beating myocyte by digital imaging microscopy. METHODS: Indo-1 was selected as a fluorescent calcium indicator. The particular feature of our digital imaging microscopy is that, in order to avoid cell movement artifacts, two indo-1 fluorescence images of different wavelengths are obtained at the same time at 16.7 ms intervals using two SIT cameras. Isolated ventricular myocytes were used, prepared from hearts of male Sprague-Dawley rats, weighing 350 to 400 g. RESULTS: In continuously beating cells contracted by electrical field stimulation, we can visualise changes in [Ca2+]i and cell shape during contraction with pseudocolour images of the fluorescence ratios, and it was found that the onset of the calcium transient preceded that of the shortening of cell length. Both isoprenaline and ouabain enhanced the magnitudes of the calcium transient and cell contraction. Furthermore, isoprenaline shortened the half decay time of the calcium transient. CONCLUSIONS: Our data suggest that the newly developed system is useful for measuring rapid changes in [Ca2+]i in relation to cell motion, and is particularly suitable for measuring the calcium transients in a beating cardiomyocyte.

Animals

Assessing the evolution of the evoked potential average: a new color display method.

A new method to control the averaging process and to check the stability of evoked potential (EP) components is described. The principle of the method is based on the display of the consecutive averages of the EP after each stimulus in a color-coded picture. In this way the evolution of the process is visualised instead of only the last average. The colors are assigned according to the amplitudes. The resulting picture initially consists of erratic colors. However, with increasing numbers of stimuli, tracks of one color soon emerge, indicating stable components. The number of stimuli necessary to generate a stable peak and the duration of stability can easily be quantified. Details with regard to the methods of extraction of essential data and data compression to record the picture are described. The color display method was evaluated with the analysis of 18 somatosensory evoked potentials and 17 visual evoked potentials of controls and 33 EPs of 18 patients. It is shown that this method preserves essential information about the averaging process: the stability of the EP components can be quantified and consequently the optimum number of stimuli to be applied. Furthermore, 14 of the 33 patient recordings showed artifacts, which could be traced with the average-evolution method, but not from the final average alone.

Color

Destruction of a sub-population of cortical neurones by suicide transport of volkensin, a lectin from Adenia volkensii.

A method for the destruction of a sub-population of neocortical pyramidal neurones is described. The technique uses unilateral striatal injections of volkensin, a toxic lectin from Adenia volkensii, which undergoes retrograde axonal transport from the site of injection to destroy subcortically projecting pyramidal cells within the neocortex. Striatal volkensin injections produce a significant reduction in the number of large pyramidal neurones of the infragranular layer. The selectivity of the lesion was demonstrated by the preservation of glutamic acid decarboxylase mRNA positive cells, considered to be cortical interneurones within ipsilateral cortex. The binding of the serotonin 1A receptor agonist [3H]-8-hydroxy-2-(n-dipropylamino)tetralin, visualised by autoradiography, was reduced in areas showing loss of large cells, indicating that these receptors may be present on subcortically projecting pyramidal cells. Ricin, another toxin lectin, but effective as a suicide transport agent in only the peripheral nervous system, produced local striatal damage but no cortical cell loss. The selective destruction by volkensin of neocortical pyramidal neurones with subcortical projection targets should aid the neurobiological investigation of such cells. Additionally, data obtained using this technique may provide insights into the interpretation of biochemical findings in neurodegenerative disease in which pyramidal cell loss is a significant feature.

Animals

The effects of haptonomy and virtual reality, anxiety, prenatal attachment and acceptance of pregnancy in unplanned pregnancy.

AIM: Unplanned pregnancies are an important public health problem that negatively affects the health of women and babies. This study aimed to determine the effects of haptonomy and virtual reality on anxiety, prenatal attachment and acceptance of pregnancy in unplanned pregnancies. Haptonomy, a touch-based bonding technique, and virtual reality, an immersive fetal visualisation tool, were used as interventions. METHODS: The sample of this randomised controlled study consisted of 217 pregnant women (haptonomy: 73, virtual reality: 72 and control: 72) who applied to the Obstetrics and Gynaecology outpatient clinics of a public hospital in eastern Turkey between July 2020 and April 2021. For both experimental groups, four interviews were conducted with the pregnant women between the 24thand32nd gestational weeks at intervals of 7-10&#x2009;days. Data were collected on pregnancy-related anxiety, prenatal attachment and pregnancy acceptance. Group differences were analysed using appropriate statistical comparisons. RESULTS: Pregnancy-related anxiety was lower, and acceptance of pregnancy was higher in the experimental groups compared to the control group (p&#x2009;<&#x2009;.001). Prenatal attachment level was higher in the haptonomy and virtual reality groups compared to the control group (p&#x2009;<&#x2009;.001). Notably, prenatal attachment scores were significantly higher in the haptonomy group compared to the virtual reality group (p&#x2009;<&#x2009;.001). CONCLUSION: In unplanned pregnancies, imagining the baby through haptonomy and imagining the baby through virtual reality are techniques that reduce the level of anxiety related to pregnancy and increase the level of prenatal attachment and acceptance of pregnancy. Especially haptonomy showed a higher effect on prenatal attachment.

Humans

Post-formation fluorescent labelling of liposomal membranes. In vivo detection, localisation and kinetics.

A fast and simple method for the in vivo/in situ detection of liposomes is described. Utilizing lipophilic carbocyanine dyes, DiI and DiO, yellow (or red) and green fluorescent liposomes can be visualised with routinely available filters. The main advantages of the method are (i) the vesicles can be labelled after they are formed and (ii) the label does not interfere with proteins on the surface of the liposomes. Labelled liposomes were found in macrophages of spleen and liver (of mice) within 30 min of intravenous administration. In the spleen, labelled liposomes localised preferentially in the marginal zone macrophages, as confirmed by double staining with FITC-Ficoll. These data correlate well with the fact that empty or haptenated liposomes are thymus-independent antigens, and that other thymus-independent antigens are also specifically taken up by marginal zone macrophages. The immunological role of these macrophages in the processing and presentation of antigen-bearing liposomes can now be studied in more detail. Administration of high doses (1-3 mg lipid) of labelled liposomes showed that uptake occurred preferentially, but not exclusively, by marginal zone macrophages. After the marginal zone macrophages had been 'saturated', the red pulp macrophages took up the liposomes. DiI and DiO have also been successfully used for labelling lymphocytes and bacteria for in vivo homing studies. The fact that liposomes can be labelled after they have been formed is an advantage for retrospective (i.e. liposomes already in use/storage) studies in e.g. targeting of drugs by liposomes.

Affinity Labels

Autoradiographic localisation of 3H-apomorphine binding sites in rat brain.

The best experimental conditions for a selective binding of 3H-apomorphine to dopamine receptors on cryostat sections were first selected by liquid scintillation quantification of the bound radioactivity. In the corpus striatum, a specific binding occurred with a half-maximal saturation concentration of about 1 nM and a maximal capacity of 180 fmol/mg of slice protein, both values in agreement with previous binding data on either membranes or slices incubated in a physiological medium. Inhibition with domperidone was clearly biphasic, indicating two classes of sites corresponding to the D-2 and D-3 sites as previously defined on membranes. When 3H-apomorphine was used at low concentrations (0.8-1.5 nM), a condition ensuring a preferential labelling of D-2 sites, rather well contrasted autoradiographic pictures were generated. The major dopaminergic projection fields in telencephalon (caudate-putamen, nucleus accumbens, olfactory tubercles) were visualised as well as other catecholaminergic regions such as the superficial gray layer of superior colliculi. Within the striatum, differences in density of these sites were observed in three perpendicular planes and confirmed by a computer densitometric image analysis. Labelling of areas of origin of the cerebral dopaminergic neurons in substantia nigra or ventral tegmental area were also observed. When a higher concentration of 3H-apomorphine (3.5 nM) was used in the presence of domperidone, another, but autoradiographically less distinct subclass of sites (D-3 sites) was demonstrated.

Animals

Autoradiographic localization of beta-adrenoceptors in pig lung using [125I]-iodocyanopindolol.

The binding of the beta-adrenoceptor radioligand [125I]-iodocyanopindolol (I-CYP) has been studied in pig lung parenchyma and the distribution of binding sites visualised by light microscopic autoradiography. I-CYP binding was saturable (maximum binding capacity Bmax = 51 +/- 3 fmol mg-1 protein), involving sites with high affinity (dissociation constant KD = 73 +/- 10 pM). Specific I-CYP binding was displaceable both by beta-adrenoceptor agonists ((-)-isoprenaline greater than (-)-adrenaline greater than (+/-)-fenoterol greater than (-)-noradrenaline greater than (+)-isoprenaline greater than (+/-)-RO363) and antagonists ((+/-)-propranolol greater than ICI-118551 greater than atenolol), indicating a predominance of beta 2-adrenoceptors. Further analysis showed that displacement data for the beta 1-selective antagonist atenolol and the beta 2-selective antagonist ICI-118551 were fitted best to a 2 binding site model and that both beta 1- and beta 2-adrenoceptors were present in pig lung in the ratio 28:72 respectively. Autoradiographic grains were localized over tissue and were most dense over alveolar walls greater than vascular endothelium greater than vascular smooth muscle greater than bronchial smooth muscle = bronchial epithelium. Atenolol (10(-5) M) caused a 31% reduction in specific grain density over alveolar wall tissue, while a 10 fold lower concentration of ICI-118551 (10(-6) M) caused a 50% decrease. These results are consistent with binding data in pig lung parenchyma demonstrating a mixed population of beta-adrenoceptors with a predominance of the beta 2 subtype. 6 It is possible that the previously described relaxant responses of the pig lung parenchyma strip to beta-agonists, mediated via beta 2-adrenoceptors, resulted from the sum of reactivities in airway and vascular smooth muscle together with relaxation of alveolar interstitial cells.

Animals