X-ray microanalytical detection of iron in pigmentation of oral mucosa.

A patient was referred to the Charles Clifford Dental Hospital with a grey metallic pigmentation of the hard palate. Conventional histopathological examination was inconclusive, suggesting the presence of either an ephelis (freckle) of pigmentation resulting from a stainless steel upper denture. Material from the pathological specimen was dewaxed and reembedded in Spurr's resin. Examination in the TEM revealed electron dense deposits in the cytoplasm of macrophages. Energy dispersive X-ray microanalysis demonstrated that these inclusions contained iron. The results suggested that the iron was in a form similar to haemosiderin and had arisen from the steel denture.     

Association of the type II cAMP-dependent protein kinase with a human thyroid RII-anchoring protein. Cloning and characterization of the RII-binding domain.

The type II cAMP-dependent protein kinase (PKA) is localized to specific subcellular environments through binding of the dimeric regulatory subunit (RII) to anchoring proteins. Subcellular localization is likely to influence which substrates are most accessible to the catalytic subunit upon activation. We have previously shown that the RII-binding domains of four anchoring proteins contain sequences which exhibit a high probability of amphipathic helix formation (Carr, D. W., Stofko-Hahn, R. E., Fraser, I. D. C., Bishop, S. M., Acott, T. E., Brennan, R. G., and Scott J. D. (1991) J. Biol. Chem. 266, 14188-14192). In the present study we describe the cloning of a cDNA which encodes a 1015-amino acid segment of Ht 31. A synthetic peptide (Asp-Leu-Ile-Glu-Glu-Ala-Ala-Ser-Arg-Ile-Val-Asp-Ala-Val-Ile-Glu-Gln-Val -Lys-Ala-Ala-Tyr) representing residues 493-515 encompasses the minimum region of Ht 31 required for RII binding and blocks anchoring protein interaction with RII as detected by band-shift analysis. Structural analysis by circular dichroism suggests that this peptide can adopt an alpha-helical conformation. Both Ht 31 (493-515) peptide and its parent protein bind RII alpha or the type II PKA holoenzyme with high affinity. Equilibrium dialysis was used to calculate dissociation constants of 4.0 and 3.8 nM for Ht 31 peptide interaction with RII alpha and the type II PKA, respectively. A survey of nine different bovine tissues was conducted to identify RII binding proteins. Several bands were detected in each tissues using a 32P-RII overlay method. Addition of 0.4 microM Ht 31 (493-515) peptide to the reaction mixture blocked all RII binding. These data suggest that all anchoring proteins bind RII alpha at the same site as the Ht 31 peptide. The nanomolar affinity constant and the different patterns of RII-anchoring proteins in each tissue suggest that the type II alpha PKA holoenzyme may be specifically targeted to different locations in each type of cell.     

A review of techniques and results obtained in one laboratory by an integrated system of methods designed for routine clinical flow cytometric DNA analysis

Establishing flow cytometric DNA analysis as a clinical routine procedure requires adequate and proven guidelines, by which the data can be obtained and interpreted to directly influence management of the individual patient with a specific neoplasm. The present paper is intended as a contribution to such guidelines, of which only fragments are available today. We have previously described a system of methods, designed for routine flow cytometric DNA analysis. In the present status report our experience, based on approximately 18,000 samples (clinical and experimental) is summarised. Sample acquisition with fine-needle aspiration, storage at -80 degrees C, internal standardization by chicken (CRBC) and trout red blood cells (TRBC), staining with propidium iodide (PI), and analysis in the flow cytometer is recapitulated, with emphasis on previously unpublished aspects. The method of statistical analysis which has an integrating role is described in some detail. A lack of linearity between channel number and DNA content was determined experimentally, and the coefficient of variation (CV) was found to decrease with increasing channel number. The corrections in the algorithm of deconvolution made necessary by these findings are fundamental for estimating the end results. The zero point adjustment and procedures for changing from one batch of standards to another are described. A systematic approach to interpretation of DNA histograms is attempted and illustrated by data from clinical specimens of malignant lymphoma, breast cancer, small cell lung cancer, cancer of the oral cavity, and bladder cancer. Some problems are still unsolved and visual inspection is required to determine if the quality of the individual histogram is satisfactory. Inspection of the fluorescence/light scatter dot-plot provides additional information for the recognition of artifacts. The results stress that good quality DNA histograms with as small CVs as possible are important for interpretation of the data. It is essential that statistical methods are employed to extract the key end-point results. These are the number of subpopulations and their relative representation, and for each subpopulation the DNA index (DI) and the fractions of cells in the cell cycle phases. For the DNA data to have any rationally based impact on clinical decision making, it must be demonstrated that they have an independent prognostic value. Strategies for final evaluation are discussed. Multicenter trials on fresh material, to accrue quickly the number of patients necessary for firm conclusions, are suggested.     

Effects of low intensity infra-red laser irradiation on ultrastructure and proliferation of liver cells in experimental hepatitis and cirrhosis]

With the aid of light, electron transmission and scanning electron microscopy and radioautography and stereometry, the influence of low-intensive laser irradiation (LILI) (infrared) was studied in normal rat liver and in experimental cirrhosis and hepatitis. It was revealed that arsenide-gallium laser irradiation causes the change of intracellular structure. These changes show the intensification on their specific function manifest in an increase of relative volume of intracellular structures. The changes of microvessels show the activation of microcirculation. The elevation of the index of the labelled nuclei testify to increased proliferation. The similar influences of LILI on the liver ultrastructure and proliferation in hepatitis and cirrhosis are accompanied by the reduction of the pathological changes of the liver--the hepatocytes oedema, granular, vacuolar and fatty dystrophy.     

The immunostimulating properties of cholecystokinin octapeptide and its fragments]

Immunostimulating properties of cholecystokinin octapeptide (CCK-8) were evaluated in experiments on adult normal and thymectomized mice, in vitro. It was shown that CCK-8 stimulates IgM-PFC production to SRBC, but does not change the immune response to Vi-antigen. CCK-8 increases the number of Thy-I+ spleen T cells and restores thymus-dependent immune response in thymectomized mice. CCK-8 has no effect on neutrophil phagocytosis activity in vitro. The immunostimulating activity of CCK-8 is related mainly to C-terminal fragment (identical to pentagastrin tetrapeptide) since the N-terminal CCK-8 tetrapeptide displays negligible effect in all tests.     

Organization of reflex sympathetic influence on rate and force of cardiac contraction]

In acute experiments on 21 cats it was proved that the change of afferent impulse on vagus nerves by means of either freeze-block or electrostimulation of their central ends results in differential reflex influences on rhythm and force of the cardiac contractions caused by sympathetic nervous system. The cut of the lower cardiac nerves may cause 'break-up' of the observed reflex, removing or inverting its ino- or chronotropy component. The given phenomenon was revealed in the experiments with high arterial pressure and with absence of tonic chronotropy influences of the left lower cardiac nerve.     

Evoked potentials and electric resistance of the cerebral cortex under the effects of strychnine and atropine]

The effect of strychnine and atropine on the amplitude of the evoked potentials (EP) of the cerebral cortex was studied. It was shown, that both drugs increased amplitude of the EP. When we used strychnine, increased amplitude of the EP determined modification of the electrical resistance and current sources of the electrical field of the EP. Atropine did not change electrical resistance and in this case only electrical current sources determined modification of the EP.     

Multiattribute evaluation of regional cotton variety trials

Summary The Australian Cotton Cultivar Trials (ACCT) are designed to investigate various cotton [Gossypium hirsutum (L.)] lines in several locations in New South Wales and Queensland each year. If these lines are to be assessed by the simultaneous use of yield and lint quality data, then a multivariate technique applicable to three-way data is desirable. Two such techniques, the mixture maximum likelihood method of clustering and three-mode principal component analysis, are described and used to analyze these data. Applied together, the methods enhance each other's usefulness in interpreting the information on the line response patterns across the locations. The methods provide a good integration of the responses across environments of the entries for the different attributes in the trials. For instance, using yield as the sole criterion, the excellence of the namcala and coker group for quality is overlooked. The analyses point to a decision in favor of either high yields of moderate to good quality lint or moderate yield but superior lint quality. The decisions indicated by the methods confirmed the selections made by the plant breeders. The procedures provide a less subjective, relatively easy to apply and interpret analytical method of describing the patterns of performance and associations in complex multiattribute and multilocation trials. This should lead to more efficient selection among lines in such trials.     

Metabolic consequences of positive inotropy in rat and guinea-pig hearts

The metabolic and physiological responses of hearts from male rats and guinea-pigs to isoprenaline and SK&F 94120 (a phosphodiesterase III inhibitor), have been studied. Doses which gave similar chronotropic stimulation gave different inotropic responses. In both species, isoprenaline generated a greater increase in developed tension than SK&F 94120. With both drugs the inotropic response in the rat was less than than in the guinea-pig. 31P-NMR investigation of high-energy phosphate levels showed reduction in PCr concentration and an accompanying acidosis in the isoprenaline-perfused rat heart only. In both species, lactate production was stimulated by SK&F 94120 but not by isoprenaline. These results are discussed with reference to G-protein activation of ion channels and differences in Ca2+ handling by the two species.