A comparative study of attachment of human, bovine and mouse blastocysts to uterine epithelial monolayer

The in-vitro attachment of human, bovine and murine blastocyststo monolayer cultures of uterine epithelium were studied bytransmission electron microscopy. The human trophoblastic cellsintrude between uterine epithelial cells forming a multilayerduring attachment in vitro, thus resembling the intrusive typeof penetration observed in vivo. The bovine trophoblastic outgrowthresembled an epithelio-chorial attachment as the trophoblastformed an attachment plate on top of the endometrial cells withoutpenetration. In the murine attachment study, the trophoblastcells immediately displaced the uterine cells and formed contactwith the culture vessel.     

GABA-immunoreactive cells in the rat gastrointestinal epithelium

Summary Frozen sections of the corpus ventriculi, antrum pyloricum, duodenum, jejunum, ileum and colon from animals perfusion fixed with glutaraldehyde were treated with an antiserum specific for glutaraldehyde-fixed GABA and processed by the peroxidase antiperoxidase method. Semithin plastic sections from the antrum pyloricum were treated similarly. Stained cells appeared in the epithelium of all segments examined except the corpus ventriculi. The highest density of cells was observed along the major curvature of the antrum pyloricum. Here they were located in the bottom half of the gastric glands. Many of the cells showed a process extending towards the glandular lumen. No significant staining in the epithelium appeared when the antiserum was preincubated with glutaraldehyde-GABA complexes, nor when the anti-GABA serum was exchanged with anti-glycine or preimmune serum. The present findings and previous physiological data suggest that GABA may play a role in gut endocrine regulation.     

Mapping the amphetamine-evoked changes in [11C]raclopride binding in living rat using small animal PET: modulation by MAO-inhibition

The performance of small animal PET for neuroreceptor studies in a psychopharmacological challenge paradigm is not yet well-described. Therefore, we used microPET and [(11)C]raclopride to map the availability of dopamine D(2/3) receptors in brain of anesthetized rats, first in a baseline condition, and again after challenge with saline or d-amphetamine. Parametric maps of the specific binding (binding potential, pB) were calculated using a reference tissue input from cerebellum, and spatially normalized to a digitized stereotaxic coordinate system for rat brain. In volumes of interest (VOIs), the mean baseline pB (n=6) was 2.05 in dorsal striatum (caudate-putamen), and 1.34 in ventral striatum (nucleus accumbens), and did not significantly differ upon retest 2 h later. The availability of [(11)C]raclopride binding sites at baseline was 8% higher in the right striatum. Challenge with amphetamine sulfate (1 mg/kg, i.v., n=4) decreased pB by 19% in both ventral and dorsal striatum. We have earlier predicted that blockade of monoamine oxidase (MAO) should potentiate the amphetamine-evoked dopamine release, thus enhancing the displacement of [(11)C]raclopride binding in vivo. However, pretreatment of rats with pargyline hydrochloride (4 mg/kg, n=4; 20 mg/kg, n=4) 1 day prior to PET did not potentiate the amphetamine-evoked reduction in dopamine receptor availability within the extended striatum. We conclude that small animal PET can be used to investigate stimulant-induced dopamine release, but that the spatial resolution is insufficient to detect differences between relative changes in dorsal vs. ventral divisions of the rat striatum. Furthermore, the present results do not reveal potentiation of the amphetamine-evoked release of dopamine in rats with MAO inhibition.     

Sublingual Nitroglycerin Delays Arterial Wave Reflections Despite Increased Aortic “Stiffness” in Patients with Hypertension: A Doppler Echocardiography Study

Venodilatation with consequent reduction in left ventricular filling and end-diastolic wall stress is an important mechanism for the beneficial effects of nitroglycerin in ischemic heart disease and in left ventricular failure. The effects of sublingual nitroglycerin on arterial pulsatile hemodynamics are less well defined. Doppler echocardiography and the calibrated subclavian artery pulse tracing were used to assess hemodynamics in subjects with sustained arterial hypertension (n = 25) before and 5 to 10 minutes after sublingual deposition of 0.5 mg glyceryl trinitrate. Aortic characteristic impedance was calculated by averaging the modulus of the input impedance (ratio of pressure to flow) at high frequencies and by calculating the ratio of pressure and flow increments during upstroke. The pressure wave was split into forward and backward components, and the reflection coefficient (the ratio of backward to forward pressures) was calculated. Parameters of the arterial bed were estimated by using 2- and 3-element Windkessel models. Nitroglycerin delayed the return of arterial wave reflections by 17% (P =.02) and increased aortic characteristic impedance by 20% (P =. 01), but it did not influence total arterial compliance. Mean arterial pressure decreased 7% (P =.0001), but pulse pressure did not change. Stroke volume and the acceleration time of aortic root flow decreased by 13% (P =.0001) and 8% (P =.01), respectively. Cardiac output decreased 7% (P =.01), despite an increase in heart rate of 10% (P =.0001). Peripheral resistance tended to decrease (4%, P =.06). Thus, in subjects with sustained hypertension, sublingual nitroglycerin dilates peripheral, predominantly muscular arteries with a subsequent delayed return of reflected pressure waves. Reflex activation of the sympathetic nervous system with consequent increased acceleration of left ventricular ejection seems to counteract the effect of reduced mean arterial pressure (distending pressure) with respect to the "stiffness" of the aorta.     

Left ventricular volumes assessed by different new three-dimensional echocardiographic methods and ordinary biplane technique

Three-dimensional (3D) echocardiography may overcome the problems with inadequate accuracy and reproducibility of 2D volume measurements of the left ventricle. Aims: To establish the in vitro accuracy and reproducibility of two new methods for 3D echocardiographic volume determination as compared to biplane measurements. Methods: Validation of volume measurements by a multiplane 3D method was performed on asymmetric latex phantoms (n=8, true volumes 45-304 ml) using rotational acquisition of 90 image planes. Porcine agarose-filled asymmetrical left ventricles (n=7, true volumes 34 – 280 ml) were measured by the same multiplane 3D method based on images acquired by probe rotation axis perpendicular (A) and parallel (B) to the ventricular long axis. Ventricular volumes were also obtained by a simplified 3D system using only the three standard apical views (C) and by the ordinary biplane Simpson’s method (D). Results: On latex phantoms systematic deviation from true volumes by multiplane 3D was less than 2%, and 95% variability of individual measurements from this mean was ± 4,9%. For accuracy on left ventricles, systematic bias was small with all the methods (<5%), but 95% variability of individual measurements was ±9,0%, 15.4%, 18.8% and 41.3% of true volumes for methods A-D respectively. Corresponding results in the same range were obtained for inter- and intraobserver variability. Conclusion: Individual in vitro volume estimates of left ventricles are of similar quality using apical multiplane or apical triplane 3D echocardiography. Both methods were superior to the ordinary apical biplane method, but inferior to multiplane 3D method with the probe directed perpendicular to the ventricular long axis.     

Lithium clearance in chronic nephropathy

1. Lithium clearance measurements were made in 72 patients with chronic nephropathy of different aetiology and moderate to severely reduced renal function. 2. Lithium clearance was strictly correlated with glomerular filtration rate, and there was no suggestion of distal tubular reabsorption of lithium or influence of osmotic diuresis. 3. Fractional reabsorption of lithium was reduced in most patients with glomerular filtration rates below 25 ml/min. 4. Calculated fractional distal reabsorption of sodium was reduced in most patients with glomerular filtration rates below 50 ml/min. 5. Lithium clearance data were independent of whether renal disease was of primarily glomerular or tubular origin and, further, were not influenced by long-term conventional antihypertensive treatment. 6. It is concluded that, even with a reduced kidney function, the data are compatible with the suggestion that lithium clearance may be a measure of the delivery of sodium and water from the renal proximal tubule. With this assumption it was found that adjustment of the sodium excretion in chronic nephropathy initially takes place in the distal parts of the nephron (loop of Henle, distal tubule and collecting duct). With more severe impairment the proximal tubule also becomes involved in the adjustment.     

A closer look at RapidArc® radiosurgery plans using very small fields

RapidArc? has become the treatment of choice for an increasing number of treatment sites in many clinics. The extensive use of multiple subfields in RapidArc? treatments presents unique challenges, especially for small targets treated in few fractions. In this work, very small static fields and subsequently RapidArc? and conventional plans for two targets (0.4 and 9.9 cm(3)) were investigated. Doses from static fields 1-4 MLC leaves (0.25-1.00 cm) wide, and larger fields with 1-4 MLC leaves closed in their centres, were measured using the portal dosimeter-based QA system EPIQA (v?1.3) and gafchromic film. RapidArc and conventional plans for two tumours were then measured using EPIQA, gafchromic EBT2 film and the phantom-based QA system Delta4. Eclipse 8.6 and 8.9, grid spacings of 1.25 and 2.50 mm and a Varian HD linac were used. For static fields one MLC leaf wide, the dose was underestimated by Eclipse by as much as 53% (v?8.6, 2.5 mm grid). Eclipse underestimated the dose downstream from a few MLC leaves closed in the centre of a large MLC field by as much as 30%. Eclipse consistently overestimated the width of the penumbra by about 100%. For the conventional plans, there was good agreement between the calculated and measured dose for the 9.9 cm(3) PTV, but a 10% underdose was observed for the 0.4 cm(3) PTV. For the RapidArc? plans, the measured dose for the 9.9 cm(3) PTV was in good agreement with the calculated one. However, for the 0.4 cm(3) PTV about 10% overdosing was detected (Eclipse v 8.6, 2.5 mm grid spacing). EPIQA data indicated that the measured dose profiles were overmodulated compared to the calculated one. The use of small subfields, typically a few MLC leaves wide, or larger fields with one or a few MLC leaves closed in its centre can result in significant errors in the dose calculation. The detector systems used vary in their ability to detect the discrepancies. Using a smaller grid size and newer version of Eclipse reduces the discrepancies observed in this work but does not eliminate them.     

Nonfunctioning pituitary adenoma: incidence,causes of death and quality of life in relation to pituitary function

The incidence of clinically significant, nonfunctioning pituitary adenoma, requiring surgical treatment, has not been established. According to previous studies, both surgery type and subsequent radiotherapy may have an impact on quality of life (QOL), and some studies have shown increased cardiovascular mortality in patients with pituitary disease. We studied all patients with functionless, suprasellar pituitary adenoma who were operated on during the period 1985–1996 (N = 192; transsphenoidal surgery = 160, craniotomy = 32). QOL was evaluated from Short Form 36 and Major Depression Inventory questionnaires. Causes of death were obtained from the Danish Register of Causes of Death. Incidence was 5.6/mill/year. Postoperatively, 27% of the patients had normal pituitary function and 27% were panhypopituitary. Fifty three patients had died. Death from cardiovascular, cerebrovascular and malignant diseases was not increased. Overall, QOL was not different from QOL of an age- and sex matched background population. QOL was, however, impaired in patients who had undergone craniotomy as compared to patients undergoing transsphenoidal surgery. QOL was not reduced in hypopituitary patients or in patients receiving radiotherapy. The study was supported by grants from The Aarhus University Hospital Research Initiative and The Obel Family Foundation, Aalborg, Denmark     

Reproductive death of cancer cells induced by femtosecond laser pulses

PURPOSE: High intensity femtosecond (1 fs = 10(-15) s) laser pulses may, via multi-photon processes, cause reproductive cell death at wavelengths that otherwise are harmless. We study the efficacy of inducing reproductive death of cancer cells by ultraviolet (UV), visible (VIS) and near infrared (IR) femtosecond laser pulses. MATERIALS AND METHODS: Human squamous carcinoma cervical cancer cells are irradiated by femtosecond laser pulses at 800 nanometers (nm), 400 nm, 266 nm and 200 nm. The reproductive death is assessed by colony forming assay. The contribution from multi-photon processes is evaluated by comparing the cell reproduction subsequent to irradiation by collimated (low intensity) and focused (high intensity), pulsed laser beams with identical fluences. RESULTS: Suitable femtosecond pulses are capable of arresting cell reproduction at all the tested wavelengths. Irradiation at 266 nm is far more efficient than the other wavelengths, both in terms of the fluence and the absorbed dose needed to induce reproductive cell death. The collimated 800 nm beam is unable to induce reproductive cell death even at a fluence of 230 Joule/square centimeters (J/cm2). However, focused 800 nm pulses with much higher intensities, but lower fluences efficiently arrest cell reproduction, thus highlighting the dramatic effect of multi-photon processes. At the intensities used in the present work focusing the 400 nm beam improves its efficacy by an order of magnitude, whereas focusing the 266 nm beam does not improve its efficacy. CONCLUSION: Femtosecond pulses at 200, 266, 400 and 800 nm induce reproductive cell death if the intensity is sufficiently high. Multi-photon processes can improve the efficacy substantially and even result in reproductive cell death at wavelengths, where single-photon processes are harmless.