Intracellular ion exchange between cytoplasmic potassium and granule histamine, an integrated link in the histamine release machinery of mast cells

Rat peritoneal mast cells isolated by gradient centrifugation in Percoll were placed between two membrane filters in a Sartorius filter apparatus and superfused with isotonic balanced salt solutions or with deionized isotonic sucrose. Histamine was released according to ion exchange kinetics. Our explanation of the observed phenomena is as follows. The superfusion induces a flow of cytoplasmic K+ ions across the histamine-containing granules, resulting in an ion exchange K+ in equilibrium Hi+ ions at the histamine binding sites. The concomitant equimolar outflow of histamine and potassium is considered to be due to a functional interplay between two histamine pools, a release and a donor pool. As the result of the K+ in equilibrium Hi+ ion exchange at the histamine binding sites of the release pool, these sites become transiently occupied by K+ ions only to be immediately reoccupied by Hi+ ions from the donor pool. The observed equimolar outflows are consistent with a 1/1 molar ratio in the exchange between histamine and potassium ions. The essential role of cytoplasmic potassium in the histamine release mechanism is a new and important observation with possible implications not only as to histamine release in general (including so-called 'spontaneous' histamine release) but also as to the release of biogenic amines and other positively charged substances stored in granules in ionic linkage to the matrix.     

An enzyme thermistor-based assay for total and free cholesterol.

A method to evaluate the free (FC) and total cholesterol (TC) in human serum, bile and gallstone extract using an enzyme thermistor (ET)-based flow injection analysis (FIA) is presented. The cholesterol in high-density (HDL-C) and low density lipoprotein (LDL-C) have also been evaluated. A heparin functionalized Sepharose column was employed for the isolation of HDL and LDL fractions from serum. The estimation of cholesterol and its esters was based on their reaction with cholesterol oxidase (CO), cholesterol esterase (CE) and catalase (CAT). Three different enzyme columns, i.e. co-immobilized CO/CAT (column A), only CE (column B) and co-immobilized CO/CE/CAT (column C) were prepared by cross-linking the enzymes on glass beads using glutaraldehyde. Column A was used for estimating FC and column C was used for estimating total cholesterol (cholesterol plus esterified cholesterol). Column B was used as a pre-column which could be switched 'in' or 'out' in conjunction with column A for the estimation of TC or FC, respectively. A calibration between 1.0 and 8.0 mmol/l for FC and 0. 25 and 4.0 mmol/l for TC was obtained. For more than 2000 assays with the ET device a C.V. of less than 4% was obtained. The assay time was approximately 4 min per assay. The cholesterol estimations on the ET correlated well with similar estimations using a commercially available cholesterol diagnostic kit.     

An efficient cryoprobe to inactivate cortical surfaces

This paper describes a cryoprobe constructed of glass capillary tubes commonly employed for cellular recordings. It is inexpensive, it is easily and rapidly constructed and most importantly it can easily be adjusted to cool larger areas and tightly fit brain curvatures and gyri.     

Progression of Ductal Carcinoma in Situ from the Pathological Perspective

Ductal carcinoma in situ (DCIS) now represents up to 20% of breast cancer cases, yet its behaviour is still poorly understood. Morphological classifications go some way to predicting prognosis, but more sophisticated approaches are required to better tailor therapy to the individual. A number of biological molecules have been identified that appear to relate to prognosis and, in model systems, promote progression to invasive disease. Some of these, such as COX-2, provide real therapeutic opportunities, whilst other marker combinations are showing promise in categorising women according to risk. Gene expression studies have led to an emerging molecular classification of invasive breast cancer, and it is now evident that at least some of these molecular subtypes can be identified at the pre-invasive stage. The difference in frequency of these subtypes between DCIS and invasive cancer may hold clues as to the biological mechanisms underpinning disease transition. It is increasingly clear that the host microenvironment can have a major impact on disease behaviour, and as well as acting as potential predictive factors, the altered microenvironment phenotype also offers novel therapeutic opportunities.     

Quantitative ultrastructural studies of gall bladder epithelium in gall stone free subjects and patients with gall stones.

The present study aimed at a further evaluation of the role of glycoproteins in the formation of cholesterol gall stones in man. An electron microscopic morphometric study of the gall bladder epithelium was performed in six gall stone free subjects and 12 gall stone patients. Six of the gall stone patients were treated with ursodeoxycholic acid three weeks before cholecystectomy. The number and the volume density of the mucin containing secretory granules, were not significantly increased in gall stone patients compared with gall stone free subjects. Treatment with ursodeoxycholic acid did not affect the number or volume density of the secretory granules. Thus, these results do not give evidence for that the degree of cholesterol saturation influences mucin content in the gall bladder wall of man. A major new finding was that gall stone patients had a markedly reduced total lysosome area and volume density of lysosomes compared with gall stone free subjects, a finding which may be related to a decreased intracellular degradation of protein and/or mucin.     

Noradrenaline reuptake inhibition enhances the antipsychotic-like effect of raclopride and potentiates D2-blockage-induced dopamine release in the medial prefrontal cortex of the rat.

We have previously observed that addition of an alpha(2)-adrenoceptor antagonist to a selective dopamine (DA) D(2) receptor antagonist enhances the antipsychotic-like effect of the D(2) blocker and also selectively increases DA output in the medial prefrontal cortex (mPFC) in rats. These data also correlate well with previous clinical trials showing augmentation by an equivalent drug combination in schizophrenia. Since the selective noradrenaline reuptake inhibitor reboxetine was found to cause similar effects on the mesolimbocortical DA system as alpha(2)-adrenoceptor antagonists, the present study was undertaken to explore whether also reboxetine might augment the effect of the DA D(2) receptor antagonist raclopride in the same preclinical model of antipsychotic activity, the conditioned avoidance response (CAR) test. We also investigated the effect of this combination in the catalepsy test for extrapyramidal side effect liability, as well as on DA output in the mPFC and the nucleus accumbens, respectively. Reboxetine (6 mg/kg, i.p.) significantly enhanced the suppressant effect of raclopride (0.1 mg/kg, s.c.) on CAR without affecting catalepsy. Administration of raclopride (0.1 mg/kg, s.c.) to rats pretreated with reboxetine (6 mg/kg, i.p.) resulted in a greatly enhanced effect on DA output in the mPFC but not in the nucleus accumbens when compared with raclopride alone. Consequently, these results suggest that noradrenaline reuptake inhibition may provide means of augmenting the efficacy of classical D(2)-antagonists in the treatment of schizophrenia, and, in principle, to generate an atypical antipsychotic drug profile.     

A phase II trial for the optimisation of treatment position in the radiation therapy of prostate cancer

BACKGROUND: Patient immobilisation and position are important contributors to the reproducibility and accuracy of radiation therapy. In addition the choice of position can alter the external contour of the treated area and has the potential to alter the spatial relationship between internal organs. The published literature demonstrates variation in the use of the prone and supine position for prostate cancer radiation therapy. Previous investigators using different protocols for patient preparation, imaging and target volume definition have demonstrated changes in the calculated therapeutic ratio comparing the two positions. We did not use rigid immobilisation, laxatives, rectal catheters or bladder voiding and assessed if in the prone position would cause a reduction of the dose to the rectum. We performed a prospective comparison of the two positions in 26 patients to determine if the differences in the spatial relation between the rectum and the planning target volume (PTV) would impact on dose-volume histograms to organs at risk (OAR). We also determined if any such improvement might permit dose escalation. MATERIALS AND METHODS: Twenty-six patients with clinically localized prostate cancer consented to participate in this study. All patients underwent a planning CT scan in both the prone and supine treatment positions. The PTV and OAR were drawn on each set of scans by one of the investigators. The PTV included the prostate and seminal vesicles with a 1cm margin except posteriorly where this margin was reduced to 5mm. The outer circumference of the bladder, rectal wall, small bowel (when present) was drawn along with femoral heads. 3D conformal treatment plans were computed using Helax TMS version 6.1B. A 3-field treatment technique was employed with energy of 10/15 MV. The prescribed dose was 70 Gy and the PTV was encompassed by the 95% isodose and the maximum dose was always less than 107%. Cumulative dose-volume histograms were calculated for the PTV, rectum, bladder, femoral heads and small bowel (when present). These non-uniform histograms for both the prone and supine treatment positions were transformed into uniform ones using the effective volume method [Kutcher J, Burman C. Calculation of probability factors for non-uniform normal tissue irradiation: the effective volume method. Med Phys 1987;14:487]. RESULTS: Twenty-one of the 26 (80%) patients had a lower effective volume of rectum irradiated if the prone instead of the supine treatment position was used. The median value of the effective volume in the supine treatment position was 31.74 Gy while the median value in the prone position was 22.48 Gy. The dose escalation was applied to the patients in the prone treatment position until the effective volume for the rectum was the same as that in the supine position. The range of dose escalation possible for these patients was 0.1-7.9 Gy. These patients could potentially have the dose escalated from the prescribed dose of 70 Gy for the supine position without any increase in side effects. For the five patients where no potential benefit was found when changing treatment position, only two patients displayed a significant (>1 Gy) advantage for the supine treatment position. Twenty-one of the 26 patients also showed an advantage for the prone treatment position in relation to bladder dose. CONCLUSION: The use of the prone position reduced the dose to the unprepared rectum and unvoided bladder in the majority of cases. It should be considered particularly in cases where large posterior seminal vesicles cause significant overlap between the planning target volume and the rectum.     

THE EFFECT OF OPERATIVE INTERFERENCE WITH THE CERVICAL SYMPATHETIC NERVOUS SYSTEM UPON THE GROWTH AND MALIGNANCY OF A TRANSPLANTABLE NEOPLASM OF THE RABBIT

The effect of the removal of the complete cervical sympathetic nervous system, of both superior and of both inferior cervical ganglia, and of a small portion of the cervical sympathetic nerve in the rabbit was studied in relation to the character of a malignant disease induced by a transplantable neoplasm. It was found that the general character of the disease which developed in the operated groups of animals was more severe than that of a similar sized control group. Comparisons of the mortality rate in the several groups, of the animal inddence of metastases, the number of metastatic foci, and the distribution of these secondary growths all showed this to be the case. There appeared, furthermore, to be differences in malignancy among the operated groups themselves. The most severe disease occurred in the group in which a portion of the sympathetic nerve only was removed (sympathotomy); that in the complete sympathectomy and superior sympathectomy groups was slightly less malignant; and that in the inferior group was much less so. These results have been interpreted as due to a less effective animal resistance, the mechanism of which has been interfered with in some way by the interference with the sympathetic nerves. The reasons for the difference in malignancy exhibited by the several operated groups are undetermined. A tentative explanation is suggested upon the basis of coordinating, favorable or deleterious functions subserved by the cervical sympathetic nervous system.