Preparation of pure neuronal and non-neuronal cultures from embryonic chick sympathetic ganglia: a new method based on both differential cell adhesiveness and the formation of homotypic neuronal aggregates. A new method has been developed for the preparation of essentially pure primary cultures of neurons and non-neuronal cells from 11-day embryonic chick sympathetic ganglia. This method utilizes (1) differences in cell-to-substrate adhesiveness between neurons and non-neuronal cells and (2) the capacity of neurons to form homotypic aggragates. The maximum difference in adhesiveness between neuronal and non-neuronal cells occurred when the ganglia were dissociated with trypsin following collection in a salt solution lacking divalent cations. This difference allowed the preparation of highly purified non-neuronal cultures and 85-90% pure neuronal cultures. Intermittent agitation during the period of cell separation markedly increased the purity of the neuronal cultures by (1) inhibiting neuronal but not non-neuronal cell attachment and (2) facilitating the formation of homotypic neuronal aggregates in the supernatant. Neuronal and non-neuronal cultures prepared under these conditions were more than 99% pure on the basis of both morphological and biochemical analyses. Both cell types exhibited attachment efficiencies greater than 95% and have been maintained for several weeks in vitro. Thus, completely isolated neuronal and non-neuronal cultures can be prepared and maintained for prolonged periods in the absence of cells of the other type.


Effects of substance P on functionally identified units in cat spinal cord. A correlation was made between the effects of synthetic substance P (ssP) on spinal dorsal horn neurones and the responses of these neurones to natural peripheral stimulation. It was found that ssP caused excitation only of those units which were excited by noxious radiant heat applied to the skin. ssP also caused a small facilitation of the excitatory response to this noxious stimulus and, in two cases, led to a response to noxious heat of units which had previously been unaffected by this stimulus. The ratio of ssP sensitive to insensitive units was highest in the deeper parts of the dorsal horn. Excitation by ssP showed a positive correlation to excitation by bradykinin. These results suggest that substance P may be involved in excitatory spinal processes and that its actions may be associated specifically with nociception.


Neuronal stimulation of (3H)thymidine incorporation by primary cultures of highly purified non-neuronal cells. A specific intercellular interaction has been demonstrated between neuronal and non-neuronal cells that appears to increase the rate of non-neuronal cell proliferation. Isolated and recombined primary cultures of both cell types were prepared from 11-day embryonic chick sympathetic ganglia by a method recently developed in this laboratory. When non-dividing neurons were added to an equal number of proliferating non-neuronal cells, the amount of [methyl-3H]thymidine incorporated by these mixed cultures was 230% greater than that incorporated by 99% pure non-neuronal cultures. Removal of all neurons from such non-neuronal cultures by a 48-h preincubation without nerve growth factor resulted in an even greater increase in [3H]thymidine incorporation upon addition of neurons (370%). When increasing numbers of isolated neurons were added to non-neuronal cell cultures, the amount of [3H]thymidine incorporation initially increased in a dose-dependent fashion until it reached a plateau. In contrast, the addition of increasing numbers of non-neuronal cells to a constant number of neurons resulted in a linear increase in [3H]thymidine incorporation. In some cases neurons and non-neuronal cells were not grown in direct physical contact but were only allowed to communicate with one another through the culture medium. Such indirect communication never resulted in a stimulation of [3H]thymidine incorporation. When neurons were added to cultures of embryonic chick fibroblasts, the neurons grew well but did not stimulate [3H]thymidine incorporation by the fibroblasts. These results suggest that embryonic sympathetic neurons selectively stimulate the proliferation of non-neuronal cells derived from the same source.


Alteration of the pattern of hippocampal nerve cell RNA labelling during training in rats. RNA synthesis in the CA3 region of brain hippocampus was studied in rats trained to a handedness reversal task. The newly synthesized RNA, labelled by intraventricular injection of radioactive orotic acid, was extracted and analyzed by micromethods. The result was following. In the trained animals the incorporation of the labelled precursor into RNA, corrected by the pool radioactivity, was almost double in comparison with the controls. In the trained animals there was a stimulation of the incorporation in the high molecular weight (greater than 18S) RNA region and in two specific low molecular weight regions: 8-9 and 16-17S. Taking in account previous results of other authors, these data appear to imply a stimulation of brain mRNA synthesis as a result of training.


Effects of fructose-1,6-diphosphate administration on learning efficiency and time sense of the honey bee, Apis mellifica carnica. In discrimination experiments honey bees had to decide between two glass dishes, one placed on a black cross, the other 40 cm away on a black square. In spontaneous selections the bees in 70% of all cases chose the black cross. The bees were then trained to search back and forth between the cross and the square until after 5 min sugar-water was given on the black square. Over a total of 5 learning trials, during which multiple decisions had been made by the searching insects, 96% of all choices were in favour of the black square. If 80 mM fructose-1,6-diphosphate (F-1,6-P2) in 1 M glucose had been ingested by the bees 30 min before the first spontaneous choice, learning was facilitated with 83% deciding in favour of the black square at the first trial compared with 46% of the controls (which had been fed on 1 M glucose only). In both groups of bees the same number of flight approches was made to the cross or square as made by the bees searching for 5 min before the reward was given. Thus no change in general activity or harvesting motivation appears to be induced by F-1,6-P2. When F-1,6-P2 had been given at the end of a successful learning series the bees favoured the black square for up to 3 days. In contrast to the controls no new incentive needed to be given during this period. Ingestion of other metabolites proved either ineffective, as in the case of fructose-6-phosphate or 5'-adenosine monophosphate, or, as observed after feeding citrate plus 3-phosphoglycerate, even reduced the performance. To test the effect of F-1,6P2 on the same time sense (circadian rhythm), bees were trained on three successive days to visit a feeding place at a specific time of day. The control bees which ingested only glucose on the evening of the 3rd day returned at their entrained 24-h interval on the 4th day. In contrast, the maximum frequency of appearance of bees fed on F-1,6-P2 was advanced by one hour, with minor appearance peaks at earlier hours of the day.


Monoamine metabolism in the developing rat brain and effects of ionizing radiation. Neonatal X-radiation induces profound changes in monoamine metabolism in the developing CNS. NE and 5-HT concentrations increase 7 days post-radiation in all CNS regions undergoing rapid axonal growth and proliferation, but not in the region of the cell bodies from which the respective neurotransmitters originate. The increase in NE and 5-HT levels is accompanied by a concomitant increase in the rate of synthesis. While these changes are evident as late as 22 days of age, the monoaminergic systems revert to normal by maturity. It is suggested that these alterations reflect an imbalance in the density of nerve endings to the region where these terminate. These regions are immature at birth and cell proliferation, a process which is affected by X-radiation, is still occurring at the time of exposure.


Isolation of synaptic vesicles from Narcine brasiliensis electric organ: some influences on release of vesicular acetylcholine and ATP. A simple density gradient centrifugation technique for separating electric organ cholinergic synaptic vesicles from other organelles and membrane fragments is described. Frozen, ground electric organ is extracted with a solution of similar density to the vesicles; during the subsequent centrifugation, vesicles remain suspended in the extraction medium and heavier contaminating structures sediment out. In confirmation of results obtained with mammalian central nervous system vesicles, a biphasic pattern of efflux of bound ACh is demonstrated. Low levels of phospholipase A2 (EC 3.1.1.4.) induce efflux of ACh from the vesicle fraction; it is shown that the concomitant fall in vesicle bound ATP is due to ATP efflux rather than ATP hydrolysis within the vesicle.


Phosphorus deficiency, parathyroid hormone and bone resorption in the growing rat. Sham-operated and parathyroidectomized (PTX) rats were divided into two pair-fed groups, one on a normal mineral intake (0.5% Ca, 0.3% P), the other on a regimen low in phosphorus (0.5% Ca, 0.03% P). P depletion led to a drop in plasma P and urine P, a rise in plasma Ca and a marked rise in urine Ca, a drop in serum magnesium and a rise in urine Mg. The changes were more pronounced in the PTX animals, but final values were the same in both groups. Parallel bone-seeking isotope (85Sr, 177Lu, 237Np) studies in nonablated animals revealed an increase in the urinary nuclide output and in the urine/tibia ratio in P-deficient animals. Normal and primary bone osteocytes decreased and enlarged osteocytes increased as a result of P deficiency; osteoclasts and osteoblasts also increased. Bone composition showed a drop in ash content and a rise in water, with a light decrease in both Ca and P, and a corresponding rise in hydroxyproline and nitrogen in the P-deficient animals. The results are interpreted to mean that P-deficiency in the young growing rat leads to an increase in bone resorption which occurs also in the absence of parathyroid hormone (PTH). The fact that final values were similar in the control and PTX P-deficient animals suggests that steady-state regulation can also occur without PTH. Because P-deficiency leads to rapid hypercalcemia and rapid marked hypercalciuria, there may exist a mechanism for phosphate regulation which would then supersede Ca homeostasis. The change in serum and urine Mg levels may reflect a decrease in tubular Ca and Mg reabsorption associated with P-deficiency.


Maintenance by cortisone of the calcemic response to parathyroid extract of rats on a diet without vitamin D and low in calcium. Weanling rats raised for 21 days on a vitamin D deprived, low Ca diet (0.02%), and given chronic cortisone administration (5 mg/kg/d), maintain responsiveness to the hypercalcemic effect of endogenous and exogenous parathyroid extract (PTE). The PTE action is a bone effect that does not require the presence of the kidneys, and is not related to a higher concentration of calcium or cortisone. Maintained sensitivity of D- Ca- Cort+ rats to PTE does not appear to be the consequence of a lesser degree of D-deficiency: the whole body vitamin D pool and its chloroform-soluble fraction in these animals are not different from those of their D- Ca- PTE- unresponsive controls. Repeated PTE injections for 4 days exhaust the sensitivity to the hypercalcemic action of PTE of D- Ca- Cort+ rats. The present data seem to indicate that cortisone-treated D- Ca- rats, responsive to the bone action of PTE, are characterized by a near normal bone calcium content and Ca/P ratio, and a significant increase in the number of osteoclasts.


Assay of bone resorption in vivo with 3H-tetracycline. 3H-Tetracycline (3H-TC) was used to quantify resorption in whole bones of growing rats and dogs. After repeated isotopic labeling of actively growing embryos or neonates, 3H-TC was observed to be distributed homogeneously and in equilibrium with 45Ca. A rapid and large loss of 3H-TC and a small loss of 45Ca occurred during the early weeks of rapid bone growth, suggesting that absolute amounts of 45Ca resorbed from bone, as reflected by losses of 3H-TC, are five to ten times greater than the net amounts of 45Ca lost from bone. Minimal loss of 3H-TC occurred due to nonspecific physicochemical exchange in vivo or in vitro (5%) except with nonradioactive tetracycline, and 3H-TC was not greatly exchanged or reused (10%) in vivo. The data are considered in terms of local and systemic conservation of calcium.


The effect of magnesium and fluoride on nephrocalcinosis and aortic calcification in rats given high sucrose diets with added phospnates. The study was conducted to observe in rats the possible modification of ectopic calcification by magnesium-orthophosphate-fluoride combinations, used as additives of diet for reduction of the cariogenicity of the sucrose. In rats, fed low magnesium diets, extra dietary orthophosphate (2%) considerably elevated the calcification of kidneys. Further additions of magnesium and fluoride partially reduced this adverse effect of phosphate. While the calcium content of the aorta in rats, fed low magnesium-high phosphate diet, was considerably elevated, the further addition of magnesium (40 ppm) partially reduced the calcifying effect of phosphate in aorta. Fluoride (15 ppm) together with magnesium (40 ppm) completely reduced it. The appearance of renal calculi caused by a low magnesium diet or by extra phosphate were similar according to light and electron microscopy except for the larger size in the latter case and occasional extratubular calculi found in groups with high phosphate-low magnesium and high phosphate with added magnesium diets.


The distribution of water in calcified turkey leg tendon. X-ray diffraction and water sorption data are presented which show that the extracellular water in calcified turkey leg tendon is associated principally with the collagen component.


Bone resorption measurement with unusual bone markers: critical evaluation of the method in phosphorus-deficient and calcium-deficient growing rats. An in vivo method to evaluate bone resorption in rats, by using unusual bone seekers not dependent on renal tubular transfer, is described and a critical evaluation of the method is made. In our experimental conditions, 85Sr and 177Lu are virtually exclusively localized in bone whereas 237Np remains unchanged in different soft organs, so that the concomitant use of these markers can be used for measuring bone resorption. If osteolysis occurs 21 days after the injection of these markers, under our experimental conditions, any increase in the urinary excretion of 177Lu and 237Np represents a rise in bone resorption, whereas an increase in Sr excretion reflects both and renal tubular events. According to our bone localization studies, the enhancement of Lu and Np excretion reflects primarily an increase in cortical bone resorption localized at the endosteal (Lu) and at the periosteal (Np) surfaces respectively. In addition, strontium is considered to be the marker of mineral resorption whereas Lu and Np, under our experimental conditions, would reflect the organic bone resorption. This method is tested in phosphorus-deficient rats and in calcium-deficient rats which exhibit disturbances of calcium metabolism at both the bone and kidney levels. In agreement with previous investigations, the use of these bone markers to evaluate osteolysis shows: (a) after a 1-week phosphorus deficiency, a slight increase in cortical bone resorption with a simultaneous fall in calcium and strontium renal tubular reabsorption, and (b) after a 1-week calcium deficiency, a high rise in cortical bone resorption with a simultaneous increase in the renal tubular reabsorption of calcium and strontium.


Remineralization of dentin in vitro.

Remineralization of dentin in vitro. The remineralization of completely demineralized bovine dentin was studied in vitro by monitoring the velocities of fall of small particles through calcifying solutions. The density of a particle of dentin may be found from its velocity of fall through a fluid using Stokes' law. The minimum concentration product of calcium and acid phosphate ions of the solution in which remineralization would take place was 3.6 (mM)2 in the presence of 22 mM bicarbonate, pH 7.35, and ionic strength 0.1. This just above the solubility product of brushite (CaHPO4-2H2O). Incubation of decalcified dentin in a phosphoprotein removed from dentin during demineralization, or in phosvitin, had no effect on remineralization. The rates of remineralization and of the fraction remineralized were inversely proportional to particle size. This inverse correlation may be due to deposition of mineral in a surface layer of constant depth irrespective of particle size. The fraction of a particle remineralized was greatly increased by the use of highly supersaturated calcifying solutions or by the incorporation of fluoride into the solutions. The empirical reaction order of remineralization for both calcium and phosphate ions was found to be unity, which is, within the error limits, equal to the order of growth of seed crystals of hydroxyapatite in calcifying solutions of the same composition.


Bone in dermatosparaxis. I. Morphologic analysis. Normal (N-) calf bone consists of lamellae regularly spaced and oriented parallel to the periosteum. The lamellae increase in thickness from the periosteum to the medullary cavity, by apposition of layers of cells and a calcifying matrix on either side of a hypercalcified primer. In the dermatosparactic (D-) bone, the hypercalcified primer is barely visible and the cells are irregularly arranged within the lamellae. The poorly defined vascular spaces are partly filled with an acellular calcified material. In the D-bone, the collagen fibers are sparse and radiate from the vascular space, while in the N-bone they are abundant and laid down concentric with the blood channels. In the D-bone, only a few weeks old, the outer lamellae are radially oriented with respect to the medullar cavity, while haversian remodeling already occurs in the inner part of the diaphysis. At 6 months, the inner half of the diaphysis is made up of normal haversian secondary bone, while the outer half is made up of radial lamellae. The alteration of the mechanical properties of procollagen fibers in the D-bone might be responsible for its defective organization. A resistant fibrous framework, therefore, seems required to ensure the spatial organization of the cells in the calcifying matrix and to maintain its cohesion.


Bone in dermatosparaxis. II. Chemical analysis. Physical and chemical analysis of the organic matrix of dermatosparactic (D-) bone was performed to evaluate the implication of an alteration in the mechanical properties of procollagen fibers in the process of calcification. The D-diaphyseal bone contained less collagen and more hydroxyapatite than normal (N-) calf bone. Fractionated by centrifugation in a density gradient, D-bone was found to be composed of more than 80% weight of a highly calcified fraction while N-bone of the same age contained only 37% of this fraction. As compared to N-bone, the amount of organic matrix per unit volume of D-bone was increased in the least calcified fractions and reduced in the most mineralized ones. The newly formed matrix of D-bone contained an amount of collagen similar to the N-bone while the concentration of other glycoproteins was higher. Calcification was accompanied, in both types of bone, by a progressive reduction of glycoproteins and collagen; the diminution of collagen was higher in D- than in N-bone. The presence in D-bone of procollagen fibers with altered mechanical properties might be responsible for an excess of glycoproteins in the newly formed matrix. Calcium salt deposition in this matrix and progressive distension of the fibrous framework are two possible mechanisms by which hypercalcification of D-bone occurs.


Scanning electron microscopy of dental calculus. The morphologic structure of anorganic dental calculus was studied by means of the scanning electron microscope. From surface observations, calculus is apparently composed of two components with distinguishable patters of calcification. One component is formed by the precipitation of minute calcific crystals on microorganisms and intermicrobial substances (plaque matrix). Such calcified masses, often spherical in shape, have a sponge-like appearance with empty spaces representing the former sites of entombed and degenerated organisms. Thus, intracellular calcification is not evident at this stage of calculus development. The other component, although having at least one common calcification front with the former, does not appear to be directly associated with microbial calcification. It exhibits a configuration of generally larger crystal growths of varying shapes and sizes. These two calcification patterns are comparable, both in distribution and size, to what has been observed by means of the transmission electron microscope, and what Schroeder has designated as "types A & B centers of mineralization," respectively. The calcific precipitation in type A centers have been identified by X-ray diffraction as hydroxyapatite. It is, therefore, speculated that the crystal patters in type B centers might represent other known forms of calcium phosphates present in calculus, such as octacalcium phosphate, whitlockite and brushite.


Calcification of alpha-elastin coacervates: a bulk property of elastin. Scanning electron microscopy and electron probe microanalysis studies are reported on thin sections of calcified coacervates of alpha-elastin. It is found that the capacity of elastin coacervates to initiate calcification is a bulk property of the coacervate and not limited to the serum-coacervate interface, that the calcium phosphate deposits act to bind the protein units together and slow the dissolution and spreading of the coacervate as it floats on an airwater interface, and that, within the limits of detectability, there is no involvement of sulfur. As the charged groups of alpha-elastin had been blocked, the initiation of deposition is due to neutral sites in the protein which are tightly bound to the calcium phosphate deposits.


Separation of potato spindle tuber viroid ribonucleic acid from Scopolia sinensis into three infectious forms and the purification and oligonucleotide pattern of fraction II RNA. Part IX. The existence of three infectious forms of potato spindle tuber viroid (PSTV) RNA from Scopolia sinensis was demonstrated by fractionation with high salt, by reverse phase and high pressure liquid  chromatography, and by polyacrylamide gel electrophoresis. Purification of fraction II was achieved by the following steps: extraction of nucleic acid with phenol, precipitation of the RNA with cetyltrimethylammonium bromide, fractionation of the RNA with lithium chloride and isopropanol, and finally gel electrophoresis. A procedure using reverse phase chromatography was developed to obtain 70-90% recovery of RNA from polyacrylamide gels. Purified PSTV fraction II RNA was digested with ribonuclease A and T and labelled with [gamma-32P[ATP using polynucleotide kinase. The labelled digests were separated by the electrophoresis-homochromatography procedures of Sanger. About 20 and 30 spots were obtained with ribonuclease A and T, respectively.


The endogenous biosynthesis of prostaglandins by brain tissue in vitro. The capacity for biosynthesis of prostaglandin F2alpha (PGF2alpha) and prostaglandin E2 (PGE2) from endogenous precursors by brain tissue slices and homogenates was measured by a gas chromatography - mass fragmentography method using deuterated prostaglandins as internal standards.  Mean biosynthesis in rat cerebral cortex slices incubated for 60 min was 60.2 ng PGF2alpha and 17.4 ng PGE2 per 100 mg of tissue. The corresponding values for homogenates were 78.1 ng and 28.9 ng. Synthetic capacity of cat cerebral cortex was considerably greater but that of human tissue was smaller than that found in rat brain. Cat cerebellum in contrast to other regions synthesized more PGE2 than PGF2alpha. The time-course of prostaglandin formation in slices was linear for the initial 60 min. Catabolism in cerebral tissues was found to be very small. Prostaglandins formed or added to the incubation media distributed between tissue and medium in a manner indicating some specific binding as well as nonspecific solubilization in tissue lipids. Norepinephrine, 3,4-L-dihydroxyphenylalanine, dopamine, adrenochrome and apomorphine greatly stimulated PGF2alpha formation probably through a nonenzymatic reduction of endoperoxides. Norepinephrine added to homogenates appeared to stabilize the fatty acid cyclo-oxygenase preventing it from inactivation. Indomethacin and Ketoprofen were potent inhibitors of biosynthesis. Paracetamol was found to be a less potent synthetase inhibitor than aspirin. The biosynthetic capacity of brain tissue in vitro appears to be orders of magnitude more than that of normal brain in situ.


Covalent assembly of mouse immunoglobulin G subclasses in vitro: application of a theoretical model for interchain disulfide bond formation. The pathways and kinetics of interchain disulfide bond formation have been determined in vitro for purified myeloma proteins representing the three major subclasses of mouse immunoglobulin G(IgG) using the reoxidation system described previously (Petersen, J.G.L. & Dorrington, K.J. (1974) J. Biol. Chem. 249, 5633-5641). Mixtures of oxidized and reduced glutathione were added to act as a disulfide interchange catalyst. The pathways of covalent assembly observed in vitro were qualitatively and quantitatively similar to those followed by the various subclasses in vivo. HH and HHL were the principle covalent intermediates seen with IgG1 (MOPC 31C) and IgG2a (MOPC 173 and clone 19). With IgG2b( MPC 11C), HL, HH and HHL were all prominant intermediates. The time courses of reoxidation were simulated using a theoretical model based on second-order reaction kinetics (Percy, J.R., Percy, M.E. & Dorrington, K.J. (1974) J. Biol. Chem. 250, 2398-2400). Two distinct phases were apparent in the reoxidation sequence. The first, which lasted for the initial 5-15 min, did not confirm to the theoretical model. The second phase could be accounted for by the model and represented the remainder of the covalent assembly process. The physico-chemical basis for this biphasic phenomenon was explored. Sedimentation velocity studies showed that noncovalent association was incomplete at the beginning of the reoxidation step for all proteins except IgG2b (MOPC 11C). No dissociation was apparent in the reduced and alkylated proteins at pH 5 in the absence of prior exposure to acid conditions. Thus, exposure to acid appears to affect the affinity between the subunits in the native proteins. Transfer of the proteins from pH 5 to pH 8.2 (the pH at which reoxidation proceeds) is accompanied by the generation of an absorption difference spectrum over an 8-10 min period. These data suggest that a pH-dependent conformational relaxation process may influence the early stages of reoxidation.


Assembly of three major subclasses of mouse immunoglobulin G: a theoretical model for covalent assembly in vivo. A mathematical model, based on second-order reaction kinetics, has been used to describe the covalent assembly of immunoglobulin G(IgG) in vitro from its heavy (H) and light (L) chains (Percy, M.E., Baumal, R., Dorrington, K.J. & Percy, J. (1976) Can. J. Biochem. 54, 675-687). In the present paper, the same model has now been applied to the steady-state assembly of IgG in vivo. This mathematical approach permits a quantitative comparison of the pathways of covalent assembly used by given immunoglobulins in vivo and in vitro. The assumptions in the model are: the species L, H, HL, HH, HHL and LHHL belong to a common pool; incompleted IgG intermediates may freely assemble to form HL, HH, HHL and LHHL; the reaction rate for covalent linkage between any two reacting species is proportional to the products of the number densities of the reactants and to a parameter P which takes the value PHH if the reaction joins two H chains, and PHL if it joins an H and L chain. In vivo values of PHH/PHL were determined for the 18 mouse myeloma tumours and cell lines studied by Baumal et al. (Baumal, R., Potter, M. & Scharff, M. (1971) J. Exp. Med. 134, 1316-1334). From these analyses, we have arrived at the following conclusions: (1) the three major IgG subclasses have distinctive values of PHH/PHL (mean value 53 for IgG1, 12 for IgG2a and 2.8 for IgG2b); (2) for IgGs of the same subclass, the values of PHH/PHL are similar; (3) the mean in vivo values of PHH/PHL are very close to those determined from in vitro assembly experiments. Finally, the individual values of PHH/PHL have been used to simulate pulse-chase experiments in the various tumours and cell lines. Considering the sources and magnitude of experimental error, the theoretical pathways of assembly agree with those determined qualitatively from the pulse-chase experiments.


The binding of phytohemagglutinin M to rat spleen lymphocytes. Quantitative studies. Phytohemagglutinin M (PHAM) has been purified from the commercial mixture of proteins produced by Phaseolus vulgaris, using a Sepharose-thyroglobulin column. The protein gave one band on gel electrophoresis and two bands on SDS-gel electrophoresis (mol. wt. 33 700 and 32 100, respectively). Molecular weight determination by ultracentrifugation gave a value of 61 200 +/- 700. The protein had a minimum sugar content of 16%. Binding studies of PHAM to purified rat spleen lymphocytes have been performed at 0, 25, and 37 degrees C. It was shown that the cells bound about the same amount of lectin at 0 and 37 degrees C, but less protein was bound at 25 degrees C. The binding phenomenon showed saturability at all temperatures. Data were analyzed by Scatchard plots and two kinds of binding sites were found. High-affinity sites and low-affinity sites have been characterized in terms of association constants and (apparent) number. It was also shown that cells treated with trypsin or sodium azide bound less lectin. Bound concanavalin A did not appear to affect the amount of bound PHAM, but its influence was reflected in the value of the association constant for the binding of PHAM. Unlabelled PHAM was shown to displace radioactive PHAM from the cells, but could not remove bound concanavalin A. The significance of these results is discussed in terms of the fluid plasma membrane model and cellular metabolism.


Differential effects of pyridoxal 5'-phosphate on lysine residues in rabbit and sturgeon muscle aldolases. Pyridoxal 5'-phosphate (PLP) in aqueous solutions can form a Schiff base complex with 14 and 16 lysine residues of rabbit and sturgeon muscle aldolases (EC 4.1.2.13), respectively. Although the mechanism of their interaction with PLP should be the same, these residues can be differentiated into three families on the basis of their inhibition constant Ki and rate constant k. The lysine residues of one of these families do not react with PLP in the presence of the substrates. Therefore, they are assumed to be part of the active center. In the sturgeon muscle aldolase, 3.7 substrate protected lysine residues are present. Rabbit aldolase, although tetrameric, contains only 2.8 substrate protected lysine residues. This suggests that one active center of this enzyme may be 'buried'. Structural studies showed the following sequence around the substrate protected lysine residues, in the rabbit aldolase: Gly-(Gly2, Val3)-Pyridoxyl Lys-Ile-Asp-Lys.


Characterization of angiotensin receptors in rabbit isolated atria. Rabbit isolated left atria have been used to study the inotropic action of angiotensin II (ATII). The peptide is active at doses ranging from 1.0 x 10(-9) to 2.8 x 10(-6)M and its inotropic effect is not modified by sotalol, phentolamine, burimamide, and indomethacin. We therefore propose that this effect results from the stimulation of receptors specific for ATII. Dose-response curves of ATII obtained in presence of increasing concentrations of 8-Gly-ATII are gradually displaced to the right, but high doses of the antagonist depressed the maximum contractions caused by ATII. pA2 value for 8-Gly-ATII in this preparation is similar to those observed in vascular and intestinal smooth muscles. Order of potency of analogues of ATII (2-, 3- and 5-Ala-ATII), acting as full agonists, but with reduced affinity, is similar to that found in rabbit aorta strips. It is therefore proposed that receptors for ATII in rabbit isolated left atria are pharmacologically similar to those present in vascular smooth muscles. Positive inotropic effects of angiotensin I, undecapeptide (1-11), dodecapeptide (1-12) and tetradecapeptide (1-14) renin substrate are antagonized by 8-Gly-ATII in similar way as the effect of ATII. This suggests that the action of these peptides is mainly due to stimulation of receptors for ATII. The contribution of myocardial converting enzyme to the action of these peptides is discussed.


Application of a multiwire proportional chamber to the detection of axoplasmic transport. A new technique for the detection of the axoplasmic transport of beta-radioactively labelled materials is described wherein a multiwire proportional chamber is used to measure the distribution of activity along peripheral nerve fibers which are maintained in vitro. The operating principles of the chamber are described and basic construction parameters given. Potential radiolabels are discussed. Two types of studies were performed at room temperature in vitro using sciatic nerves of the amphibian Xenopus laevis: static and dynamic. In the static study the nerve ganglion was incubated for a suitable period of time in either L-(U-14C) leucine or L-(35S) methionine after which the ganglion was removed and the activity in the remaining nerve assayed with the chamber. In the dynamic study the nerve activity was assayed by the chamber while incubation proceeded so that a dynamic picture of transport could be observed. Using the second approach, transport rates were observed which are in agreement with others which have been reported in the literature. Some advantages and limitations of the technique are discussed.


Anti-inflammatory and analgesic properties of etodolic acid in rats. Etodolic acid (1, 8-diethyl-1, 3, 4, 9-tetrahydropyrano(3, 4-b) indole-1-acetic acid) showed potent anti-inflammatory and analgesic properties in rats. In adjuvant arthritic rats etodolic acid was approximately six times more potent than phenylbutazone. In the carrageenin and the analgesic assays (inflamed paw pressure test) its potency was comparable to that of phenylbutazone. The potency of etodolic acid in the adjuvant arthritic rat suggests that this novel anti-inflammatory compound will be effective in the treatment of arthritic conditions of man.


A specific sugar transport mechanism in smooth muscle and its regulation. The membrane transport of 3-O-methyl-D-glucose was studied in vitro in a smooth muscle, the detrusor of rat urinary bladder. Transport occurred by facilitated diffusion and showed the same chemical specificity and sensitivity to specific inhibitors as skeletal and cardiac muscle but its insulin sensitivity was smaller. Transport was increased by agents inhibiting the Na+pump and was decreased by agents which increased Na+ and K+ gradients by apparently stimulating the Na+pump. In accord with a rate limiting role of transport in glucose utilization, similar stimulating and inhibitory effects were seen when CO2 production from (14C) glucose was measured.


[A method to measure the rate of metabolic clearance and secretion of cortisol in the eel (Anguilla anguilla L.)]. Classical methods for the analysis of steroid dynamics were adapted to measure the metabolic clearance rate and the secretion rate of cortisol in the European eel using (1, 2-3H) cortisol as a tracer. 1. Measurements were made whilst the eels were in the steady state; stresses which usually result from anaesthesia and handling were avoided by chronic catheterization of the blood vessels of the swim bladder. In this way, the plasma cortisol level were constant; thus, it was possible to obtain valid measurements of the cortisol secretion rate knowing the metabolic clearance rate and the plasma cortisol level. 2. In the eel, cortisol and cortisone together form a chemical compartment', derived originally from cortisol secreted by the interrenal gland and subsequently transformed partially into cortisone in the plasma and maintained thenceforth in dynamic reversible equilibrium. Consequently, the metabolic clearance rate of cortisol has been calculated from the known changes in concentration of both (1, 2-3H) cortisol and (1, 2-3H)cortisone in the plasma deduced from dichloromethane-extractable plasma radioactivity, 72% of which is always attributable to these two steroids from the moment that the radioactivity begins to decline regularly as a function of time (15-30 min after the injection). 3. After a single injection of (1,2-3H) cortisol, the interpretation of the disappearance curve by compartmental analysis is complicated by the delay of tracer distribution and by the evident variability in the initial portion of the curve. Disappearance curves have accordingly been analysed in a more rigorous fashion either by integrating the area under each curve which allows one to calculate individually the metabolic clearance rates, or by integrating according to Normand, M., and Fortier, C. (1970. Can. J. Physiol. Pharmacol. 48, 274-281) which allows one statistical treatment of data. 4. Estimates of the metabolic clearance rate of cortisol were made in a satisfactory manner by either one of the two integration methods. They were also very similar to estimates obtained by prolonged infusion of (1, 2-3H) cortisol to constant specific activity into eels maintained under identical experimental conditions.


A compartmental model for bilirubin kinetics in isolated perfused rat liver. Bilirubin kinetics were studied in an isolated, perfused rat liver system using unconjugated (14C) bilirubin (UC(14C)B) and  delta-amino (4-14 C) levulinic acid (A(14 C)LA) to derive a suitable compartmental model. Plasma disappearance of UC(14C)B, plasma appearance of conjugated (14c) bilirubin (C(14C)B) and biliary excretion of C(14C)B were followed for 90-120 min following injection of UC(14C)B. Hepatic content of labeled bilirubin 12 min after the injection  of UC(14C)B was determined directly in five separate perfusion experiments. UCB was found to reflux back to plasma from liver in two experiments using A(14C)LA. Bilirubin binding to red blood cells (6-8% of the perfusate level) and the components of the perfusion apparatus (4-6% of perfusate level) was estimated by performing a control experiment without the liver. A six compartment model was necessary and adequate to explain the experimental data and current knowledge of bilirubin metabolism: (1) UCB bound to red blood cells and the perfusion apparatus, (2) plasma UCB, (3) liver UCB, (4) liver CB, (5) plasma CB, and (6) bile CB. The proposed model could serve as a reference point for studies of bilirubin kinetics in whole animals for normal and abnormal states.


Adrenergic neuron blocking action of dehydrocorydaline isolated from Corydalis bulbosa. Dehydrocorydaline, an active principle of Corydalis bulbosa alkaloids, in concentrations of 10(-5) M to 5 x 10(-5)M inhibited relaxation and the concomitant release of (3H)-noradrenaline caused by 10(-4)M nicotine and electrical perivascular nerve stimulation in the taenia caecum of guinea pig. The same inhibitory effects were observed on contraction and release of (3H) noradrenaline in the sympathetic nerve-pulmonary artery preparation of rabbit. On the other hand, neither relaxation nor contraction caused by exogenously applied noradrenaline was affected. These results suggest that the inhibitory action of dehydrocorydaline on the relaxation or contraction, produced by nicotine and electrical nerve stimulation, is due to blockade of noradrenaline release from the adrenergic nerve terminals in both the taenia caecum and pulmonary artery. Participation of the adrenergic neuron blocking action of dehydrocorydaline in preventing experimental ulceration is discussed.


Motor and sensory fibres of neck muscle nerves in the cat. The numbers and sizes of nerve fibres to the dorsal neck muscles, splenius, complexus and biventer cervicis have been examined in the cat. The total number of fibres is unusually high as is the content of sensory fibres (estimated as the loss of fibres after ganglionectomy). The fibre spectra of these sensory nerves has an unusually large number of fibres in the group II and III range (3-7 mum) and differs markedly in this way from other muscle nerves. The motor fibres contain a high proportion (64-99%) in the gamma fibre size range. Large motor fibres are absent in the nerves to biventer cervicis (a slow muscle). The ratio of unmyelinated to myelinated fibres in neck muscle nerves is similar to that in hind legs at about 2.5:1.


Further evidence that protein synthesis can be decreased in vivo following hormonal stimulation in the rat pancreas. The present study has been undertaken to determine in the rat the influence of exocrine secretory stimulation on pancreatic protein synthesis. This stimulant consisted of a single injection of cholecystokinin-pancreozymin (8 Ivy units/kg) plus secretin (5 clinical units/kg). The rate of (14C) phenylalanine incorporation into total proteins was measured 5, 11, 17, 30, 45 and 60 min later. Incorporation was significantly decreased after 5 min, then significantly increased at 17 min, and finally returned to control values at 45 min. This biphasic evolution was shown not to be caused by variations in the precursor pool specific radioactivity. We concluded that secretory stimulation of the pancreas can induce a decrease in the rate of protein biosynthesis. This decrease is nevertheless a transient phenomenon, since the rate of biosynthesis was increased at 17 min. These results, obtained from a totally in vivo system, confirm previous data obtained from an in vivo-in vitro system.


Potassium and the recovery of splenic capsular smooth muscle after cold storage. Cat spleen capsular smooth muscle, depleted of potassium and enriched with sodium by cold storage in a potassium free medium, relaxed and underwent transient reduction in responsiveness to noradrenaline when potassium was introduced into the bathing medium. Both these effects could be blocked by ouabain, lithium substitution for sodium or low ambient temperature, suggesting possible involvement of the sodium pump. In the continued presence of potassium, relaxation was maintained but sensitivity to noradrenaline increased, possibly due to restoration of normal intracellular sodium and potassium concentrations.


The effects of selenium on the metabolism of methionine in sheep. Two groups of sheep fed a diet of hay known to produce nutritional muscular dystrophy, one group of which received selenium supplementation, were used to study the effects of selenium on the metabolism of administered L-(35S) methionine by rumen microflora. Rumen bacterial proteins of the Se supplemented sheep contained significantly higher levels of radiosulfur than the bacterial protein of the non-supplemented sheep. Of hte L-(35S) methionine present in the rumen liquor samples from Se-supplemented sheep 2 h after administration, 13.3% of the amino acid, which was measured as methionine sulfone, was found in the microbial proteins. A large proportion of the administered labeled methionine was resynthesized as cyst(e)ine which may account in part for that determined as cysteic acid in rumen bacterial and plasma proteins. The observed low levels of radiosulfur found in rumen microflora from selenium deficient wethers, indicates that the presence of selenium profoundly affects the rate of methionine metabolism and the distribution of methionine in rumen bacterial and protozoal proteins. In another experiment, the effect of selenium on the metabolism of L-(Me-3H) methionine was studied. The selenium status of the sheep had no significant effect (P greater than 0.05) on the distribution of 35S radioactivity in the blood plasma and tissues.


Vascular actions of furosemide and bumetanide on the rat superior mesenteric vascular bed: interactions with prolactin and prostaglandins. The mesenteric vascular bed of the rat was used to investigate the effects of furosemide, bumetanide, prolactin, aspirin, indomethacin and prostaglandin E2 on the pressor response to norepinephrine. Furosemide, bumetanied and indomethacin could all inhibit the responses to norepinephrine: in each case responsiveness was restored by the addition of prostaglandin E2 to the perfusate. Bumetanide and furosemide both failed to inhibit responsiveness in the presence of an adequate amount (50 pg/ml) of prostaglandin E2. As has been shown previously, ovine prolactin in a concentration of 50 ng/ml enhanced pressor responses to norepinephrine while 500 ng/ml, after an initial potentiation, inhibited responsiveness. Aspirin, furosemide and bumetanide all reversed both the potentiation produced by the lower prolactin concentration and the inhibition produced by the higher one. When taken in conjunction with other evidence these results suggest that the diuretics exert their vascular effects by inhibiting prostaglandin synthesis whereas prolactin acts by stimulating such synthesis.


A method for the study of the inhibition of DNA synthesis by rabbit tissue extracts. This paper describes a method to measure DNA synthesis in the jejunum. Everted rings of rat jejunum were incubated in the presence of (3H) thymidine. The specific activity of jejunal DNA was assayed at the end of incubation. This method was found to be valid when (1) incubation time was 30 min, (2) (3H) thymidine concentration was 90 ng/ml, and (3) paired flasks containing four, respectively, contiguous rings were compared. This method was used to investigate the existence of an intestinal chalone. An aqueous extract (S-105)of rabbit small intestine mucosa was prepared and tested for DNA synthesis inhibition in jejunum. A significant inhibition was noted, which was an hyperbolic function of the quantity of extract present in the media. The study of S-105 inhibition on other organs (kidney, liver, colon) failed to demonstrate organ specificity. However, aqueous extracts from intestinal organs were found to exert a greater inhibition than aqueous extracts form other organs. It is suggested that S-105 contains other nonspecific inhibitory fractions; our data prove the necessity of an extensive purification to demonstrate the existence of a specific chalone that regulates intestinal cell proliferation.


Effect of (Sar1, Ala8)-angiotensin II and hypophysectomy on the intestinal resistance vessels and blood pressure following furosemide-induced volume depletion. Intravenous administration of furosemide (2 mg/kg) caused intestinal vasoconstriction in various groups of pentobarbital-anesthetized cats. (Sar1, Ala 8)-angiotensin II, a specific competitive antagonist of angiotensin II, was infused 60 min after administration of furosemide, a time when the intestinal vasoconstrictor response to the diuretic was maximal or near maximal. In hypophysectomized animals, infusion of the antagonist abolished the intestinal vasoconstriction and caused a significant fall in arterial pressure even when the intestinal nerves and adrenal glands remained intact. In contrast, the antagonist had little effect when the pituitary gland remained intact. The results suggest that endogenous angiotensin and vasopressin are overlapping mechanisms which constrict the intestinal resistance vessels and support arterial pressure following furosemide-induced volume depletion. In the absence of one control system, the other compensates to maintain the responses.


Duration of analgesia in mice after heroin by two testing methods. The analgestic action in mice of single injections of heroin hydrochloride ranging from 0.3-240mg/kg was measured by the tail-clip and by the hot-plate methods. The duration of analgesia increased as the dose of heroin increased. By the tail-clip technique, the mean effective dose (ED50) (and standard error of estimate) at 30 min was calculated as 1.0 (+/3.42) mg/kg while at 180 min it was 27.5 (+/3.05). By the hot-plate technique the ED50 at 30 min was 4.9 (+/3.13) mg/kg and at 180 min it was 173.8 (+/5.38) mg/kg. The hot-plate method, though less sensitive than the tail-clip method, yields a regression line derived from ED50 values at various testing times of the same slope; thus the two methods give comparable results for changes in analgesia with time. The rate of change of the median analgesic dose of heroin HC1 in mice was calculated to be 2% per minute. In similar mice the acute mean lethal dose for single, subcutaneous injections of heroin HC1 was calculated to be 190.5+/3.01 mg/kg (95% confidence limits).


The effect of metoclopramide on intestinal muscle responses and the peristaltic reflex in vitro. Metoclopramide (Mcp) is known to facilitate gastrointestinal emptying in vivo and to stimulate various isolated intestinal muscle preparations. On the guinea pig ileum, taenia coli, rabbit ileum and rat duodenum, Mcp increased the tone and responses to acetylcholine, carbachol and nicotine; had no effect on responses to histamine, potassium chloride and prostaglandin E1; decreased responses to 5-hydroxytryptamine (5-HT). Atropine, methysergide, morphine, and tetrodotoxin, alone or in combination, partially blocked the stimulatory responses to Mcp, but hexamethonium, mepyramine and indomethacin did not. Mcp (1.0 muM) lowered the threshold for elicitation of the peristaltic reflex to a sub-threshold intraluminal pressure (2.5 cm water), facilitated the peristaltic response to threshold pressures (3-4 cm water) and restored the reflex in fatigued preparations, but not that depressed by cooling to 24 degrees C. During block of peristalsis by atropine, hexamethonium or methysergide (applied serosally) 5-HT (0.25 muM) but not Mcp (1.0 muM)) effectively restored the peristaltic reflex, but neither antagonized the inhibition by morphine or procaine acting serosally. However, Mcp (1.0 muM) re-established peristalsis inhibited by a high concentration of 5-HT ((4 X 10 muM). These results do not support the hypotheses that the stimulatory action of Mcp is entirely dependent on either peripheral sensitization of muscarinic receptors or an action on tryptaminergic mechanisms but are consistent with our previous conclusion that an additional component may be a blockade of some intrinsic inhibitory (possibly purinergic) substance normally restraining intestinal motility or tone.


Brown adipose tissue-a factor in the survival of harp seal pups. The respiratory characteristics of mitochondria isolated from the subcutaneous brown adipose tissue of newborn harp seals indicate that the tissue is thermogenically active. Temperature recordings in vivo revealed, in fact, that the tissue was maintained at a temperature close to that of the body core during immersion of the pups in ice-water. Beta-adrenergic blockade markedly increased the cooling rates at both locations in ice-water, while curarization, accompanied by artificial respiration did not. We conclude that nonshivering thermogenesis through activated brown adipose tissue plays a decisive role in the defence against cold in the newborn harp seal.


Postjunctional supersensitivity of the rat vas deferens and gap junctions. Tissues from the duodenum and vas deferens of Sprague-Dawley rats were examined of the rat vas deferens and gap junctions. Can. J. Physiol. Pharmacol. 54, 412-416. by electron microscopy after glutaraldehyde fixation and postosmication. Gap junctions (nexuses) were readily demonstrated in the duodenum in both control and reserpine treated animals (1.0 mg/kg per day for 7 days). Honever, gap junctions could not be demonstrated in vas deferens. It is concluded that the postjunctional supersensitivity and spontaneous activity induced by reserpine in vas deferens, does not result from the formation of gap junctions.


Neurologic disorders responsive to folic acid therapy. Six women aged 31 to 70 years had folate deficiency and neuropsychiatric disorders. The three with acquired folate deficiency were depressed and had permanent muscular and intellectual fatigue, mild symptoms of restless legs, depressed ankle jerks, diminution of vibration sensation in the legs, stocking-type hypoesthesia and long-lasting constipation; D-xylos absorption was abnormal. The bone marrow was megaloblastic in only one patient, and she and one other had atrophy of the jejunal mucosa. The third was a vegan. All three recovered after folic acid therapy. The other three were members of a family with the restless legs syndrome, fatigability and diffuse muscular pain. One also had subacute combined degeneration of the spinal cord and kidney disease but no megaloblastosis; she improved spectacularly after receiving large daily doses of folic acid. The other two also had minor neurologic signs, controlled with 5 to 10 mg of folic acid daily. Unrecognized and treatable folate deficiency (with low serum folic acid values but normal erythrocyte folate values) may be the basis of a well defined syndrome of neurologic, psychiatric and gastroenterologic disorders, and the restless legs syndrome may represent the main clinical expression of acquired and familial (or inborn) folate deficiency in adults.


Recurrent electromechanical dissociation due to severe underlying coronary heart disease. Recurrent electromechanical dissociation, eventually fatal, presented as Stokes-Adams-like syncope in a patient with severe underlying coronary heart disease. Mechanical function was probably suppressed because of myocardial ischemia due to recent occlusion of the proximal right main coronary artery; the left main coronary artery had been occluded previously. Spasm of the proximal right coronary artery may have accounted for his recurrent attacks of pain and syncope. It is suggested that this clinical picture has a grave prognosis, demanding urgent therapeutic measures. While these measures are likely to be more hazardous than usual, the grave prognosis and potential for therapeutic success make them well worth the risk.


Methylmalonic acidemia controlled with oral administration of vitamin B12. A 3-month-old male infant had two episodes of fever, projectile vomiting, dehydration, generalized fine tremors and gross metabloic ketoacidosis. Methylmalonic acid was found in high concentration in both serum and urine, although the concentration of serum vitamin B12 was normal. A therapeutic trial of vitamin B12, administered parenterally, reduced greatly the methylmalonic aciduria. The patient has since been given vitamin B12 supplements continuously, initially 1 mg intramuscularly every other day, then 15 mg/d orally, and the protein in his diet was subsequently restricted. The most effected control of the methylmalonic aciduria was achieved with the combined regimen of oral vitamin therapy and dietary protein restriction. His physical and intellectual development have progressed normally and he has survived several acute respiratory tract infections without recurrence of metabolic acidosis.


Herpes simplex encephalitis: treatment with surgical decompression and cytosine arabinoside. Herpes simplex encephalitis in a 21-year-old man presented as a flu-like illness, followed by inappropriate behaviour, drowsiness and focal neurologic signs. Investigations indicated a lesion in the right temporal lobe. The diagnosis was confirmed by isolation of the virus from a cerebral biopsy. Pronounced clinical improvement was noted when cytosine arabinoside therapy was started in the postoperative period. This report supports the observation by some authors that cytosine arabinoside may be of value in the treatment of herpes simplex encephalitis.


Effect of physical training on hemodynamic performance following myocardial infarction: a controlled study. The effect of physical training on hemodynamic performance was evaluated in a group of patients who had had a myocardial infarction and a group of healthy, age-matched controls. Before training, the patients' mean ventilatory equivalent was significantly less than that of the controls at the lowest workload (300 kpm/min), the mean stroke volume was significantly increased at the highest workload then achieved (600 kpm/min), and the mean arteriovenous oxygen content difference was significantly smaller at the highest workload. The patients had a relative bradycardia before training and there was no significant reduction in mean resting or submaximal heart rate after training. Their mean oxygen uptake was significantly reduced at the lowest exercise workload after training and this response was significantly different from that of the controls after 8 weeks of training. Mean cardiac output during exercise was significantly reduced in the patients after training, but only at the 600-kpm/min workload, the response being blunted at 900 kpm/min; mean stroke volume was also significantly reduced at this workload after training; both these responses were significantly different from those of the controls Mean arteriovenous oxygen content difference at 6oo kpm/min was significantly increased in the patients after training, though the response was not significantly different from that of the controls. Mean ventilatory equivalent was also significantly increased in patients after training, becoming similar to that of the controls.


Psychiatric illness in the medical profession: incidence in relation to sex and field of practice. The overall incidence of psychiatric illness among the physicians of British Columbia during 1970-74 was 1.27% per year. The overall suicide rate was more than 36.5/100 000. Incidence was not dependent on sex or age. The two specialties with the highest incidence--ophthalmology and psychiatry--had previously been demonstrated to have significantly high rates of suicide. The highest incidence was among psychiatric residents; in other resident groups collectively the incidence was at the expected rate. Greater severity of illness and poorer prognosis was found in family physicians compared with specialists, although the incidence was the same in the two groups.


Sex therapy: considerations in the selection of patients. Sex therapy has proven helpful for many patients with sexual dysfunctions. In judging the appropriateness of this new treatment procedure the clinician should determine the following: that a sexual dysfunction exists, that a stable couple relationship exists, that there are no major marital, psychiatric or physical problems that are etiologically important or that would interfere with treatment, and that both partners are willing to engage in a treatment program.


Hospitalization for mental illness: evaluation of admission trends from 1941 to 1971. A study of the diagnostic composition of the inpatient population of Ontario and Canadian psychiatric facilities has shown an important change in hospital-treated illness over the period 1941-71. Patients with nonpsychotic disorders accounted for 54% of all admissions to Ontario public mental hospitals in 1971, compared with only 8% in 1941. The trend was similar for both first admissions and proportion of readmissions, and was similar for psychiatric units of general hospitals. In contrast, the overall rate of first admission for psychotic disorders to inpatient facilities remained remarkably constant over time, as did the proportion of readmissions among all admissions. The findings dispel the notion that the increasing proportion of readmissions is due largely to a rapid turnover of former long-stay psychotic patients (the "revolving-door phenomenon"). The findings could not be attributed to a changing prevalence of types of psychiatric illness, increased availability of psychiatric inpatient facilities or comprehensive medical insurance.


Endobronchial lipoma: two cases and review of the literature. Endobronchial lipoma is a benign tumour of the large bronchi occurring in middle-aged men. To the 38 successfully treated cases in the English literature a further 2 are added. The symptoms are those of obstructive pneumonitis mimicking bronchogenic carcinoma, and the result of delayed therapy may be bronchiectasis. Treatment includes local resection through a bronchoscope or a bronchotomy incision, or removal, if necessary, of the obstructed lobe or lung at thoracotomy. Smoking may be important in the pathogenesis of this tumour.


Airway obstruction following smoke inhalation. Respiratory distress with episodes of cyanosis, intercostal retraction and sibilant rhonchi occurred in a 2-year-old boy over a 48-hour period following serious smoke inhalation. Worsening of the child's condition accompanied these findings, culminating in sudden loss of air entry, severe respiratory distress and loss of consciousness, which necessitated endotracheal intubation for resuscitation. Pronounced improvement followed removal of two pseudomembranous bronchial casts from the airway by suctioning, and thereafter recovery was uneventful.


Tartrazine-containing drugs.

Tartrazine-containing drugs. Pharmaceutical manufacturers producing or distributing drugs in Canada were surveyed between December 1974 and March 1975 to determine which of their products contained tartrazine, a pyrazole aniline dye. A list of some 580 drug products of the 156 manufacturers who responded is presented for aid in managing the tartrazine-sensitive patients.


Triiodothyronine and thyroxine interrelationships in health and disease. With the recent development of radioimmunoassay techniques for the measurement of serum triiodothyronine (T3) concentration, new concepts have arisen regarding the biologic role of T3 in health and disease and its interrelationships with thyroxine (T4). An awareness of the influence of clinical conditions that affect binding of thyroid hormone to plasma proteins is required in the interpretation of moderately increased or decreased serum T3 values. Hormone preparations containing T3 may produce transient increases in T3 concentration into the hyperthyroid range. Measurements of serum T3 concentration appear to be particularly indicated in clinical situations in which hyperthyroidism is suspected but serum T3 resin uptake and serum T4 values are normal, to exclude the T3-toxicosis syndrome. Also, when serum T4 values are in the hypothyroid range, measurement of serum T3 as well as serum thyrotropin (TSH) concentrations can lead to recognition of abnormalities in thyroid gland biosynthesis. Before a diagnosis of hypothyroidism is made on the basis of a low serum T3 value, one must exclude a variety of clinical nonthyroidal conditions that can result in changes in plasma T3 protein binding or impaired peripheral conversion of T4 to metabolically active T3 without producing a hypometabolic state.


A case of cholera in Kingston, Ont.

A case of cholera in Kingston, Ont. A case of cholera occurred in Kingston, Ont. in 1974 in a traveller from South Africa. Treatment, based on an understanding of the pathophysiology of cholera diarrhea and the mechanism of action of the Vibrio cholerae enterotoxin on gastrointestinal fluid loss, consisted of correcting the severe loss of fluid and electrolytes and the metabolic acidosis, as soon as the patient could tolerate taking fluids orally, further fluid replacement consisted increasingly of oral administration of glucose and saline. Tetracycline therapy was given only to shorten the duration of the acute illness.


Laboratory investigation and infection control of cholera in Kingston, Ont. Vibrio cholerae, biotype El Tor, was isolated in a hospital laboratory in Kingston, Ont. in 1974. Confirmation and complete identification by the Ontario regional and provincial public health laboratories was obtained within 3 days. Institution of well established infection-control and public health measures prevented spread of the infection within the hospital and the community.


Cholera: synopsis of clinical aspects and principles of treatment. Cholera varies greatly in clinical severity; the mortality of untreated severe cholera may be as high as 60% The main clinical feature is dehydration; fluid lost in the stools may amount to 60/. Rehydration is the cornerstone of treatment. The amount of fluid required is approximately 10% of body weight in severe dehydration and 5 to 8% in moderate dehydration. Fluid therapy, which must be individualized, may be successful on its own, but chemo-therapy shortens the duration of illness. Tetracycline (in adults, 40 mg/kg for 2 days; in children, 50 mg/kg for 2 days) reduces the fluid loss and eliminates the causative organisms. Vaccination is of limited value.


Cardiac hemolytic anemia resolving after second mitral annuloplasty. Following an episode of rheumatic carditis, severe mitral incompetence developed in a 9-year-old girl. A mitral annuloplasty succeeded for a short time in ameliorating her symptoms of cardiac failure. However, mitral incompetence recurred and was accompanied by severe anemia and hemosiderinuria. Distortion of erythrocytes was evident on a peripheral blood smear. A second mitral annuloplasty resulted in resolution of the hemolytic anemia.


Coarctation of the aorta in adults.

Coarctation of the aorta in adults. Thirty-six patients, 19 men and 17 women, presented at age 18 or older between 1952 and 1974 with coarctation of the aorta. Of the 14 (39%) who had associated cardiovascular disease, 12 had aortic stenosis or insufficiency or both. Three patients had infections-two, endocarditis (aortic valve) and one, endarteritis. Three of the seven patients who did not undergo an operation are alive, two at more than 50 years of age. Five patients had myocardial infarctions, two at 35 years of age. Twenty-nine (80%) had operations; in eight instances the patient was over age 40. All 18 patients undergoing repair of isolated coarctation survived, while only 7 of the 11 patients with associated cardiovascular lesions who underwent repair recovered. Aortic valvular disease and myocardial infarction are serious complicating factors in coarctation of the aorta.


Design and evaluation of head unit for wheelchair control by quadriplegic patients. A control unit was designed to allow persons who have lost hand and arm function to control the speed, steering, reversal and on-off switching of an electric wheelchair by means of backward movement and rotation of the head. When possible, shoulder movement was used to control both reversal and on-off switching. Clinical evaluation in 10 patients with quadriplegia and 2 with severe neuromuscular disease showed that the unit neither interfered with nor restrained the patients' residual body movements, permitted use of natural head movements for smooth and fast control of the wheelchair, and was well accepted by and integrated into the life of the patients.


Hodgkin's disease following thorium dioxide angiography. Hodgkin's disease occurred in a 53-year-old man who, 25 years previously, had undergone cerebral angiography, for which thorium dioxide suspension (Thorotrast) was used. Deposits of throium dioxide were noted in reticuloendothelial cells in various locations. An association between thorium dioxide administration and the subsequent development of malignant tumours and neoplastic hematologic disorders has previously been reported.


Transmission of hepatitis B by a human bite: an occupational hazard. Hepatitis B developed in a policeman 15 weeks after he was bitten on the hand. A few days after the bite hepatitis B developed in the assailant. The bite had drawn blood and this method of inoculation was presumed to be the route of transmission of the virus. Compensation was awarded on the grounds that this was an occupationally acquired disease.


Scabies masquerading as Letterer-Siwe's disease. Letterer-Siwe's disease was diagnosed from clinical appearance and initial assessment of a skin biopsy in a child with a 2-month history of skin rash. Fine erythematous papules were scattered on the trunk. The biopsy showed epidermal thickening and an inflammatory infiltrate chiefly in the upper layers of the dermis; deeper in the dermis the infiltrate was perivascular and periappendicular, histiocytes predominating in some areas and lymphocytes in others. A diagnosis of scabies was made after burrows were demonstrated on palms and soles and the mite of scabies was isolated from them.


Phenylketonuria variants in Ontario.

Phenylketonuria variants in Ontario. Since mass screening of the newborn population for phenylketonuria (PKU) by the Guthrie test was begun in Ontario in July 1965 many variants of PKU have been recognized in the 96 to 97% screened. Seventy-one cases of classic PKU were detected (four were missed). Of 48 cases of persistent hyperphenylalaninemia discovered, 18 were classified as atypical PKU and 30 as persistent benign hyperphenylalaninemia. Numerous infants with transient hyperphenylalaninemia (initial values over 10 mg/dl in 12), in many instances the result of transient neonatal tyrosinemia, were discovered. There was a slight predominance of males. Serum phenylalanine values of up to 15 mg/dl seemed to be harmless to the developing brain. A survey of 67 247 adults in the general population revealed 1 person with PKU and 1 with persistent benign hyperphenylalaninemia; both had normal intelligence quotients. Of 1548 mothers of retarded children tested, none had hyperphenylalaninemia.


Removal of amantadine hydrochloride by dialysis in patients with renal insufficiency. Two patients with Parkinson's disease and renal insufficiency had excessively high concentrations of amantadine hydrochloride in the blood. The amounts of the drug removed by hemodialysis and peritoneal dialysis were small. However, since extrarenal elimination is negligible in such patients, frequently repeated dialysis may be required to remove the drug.


Restaging of breast cancer in a northern Alberta registry. All breast cancer patients presenting preoperatively at the Dr. W.W. Cross Cancer Institute, Edmonton, since 1971 are included in a registry that provides detailed clinical and epidemiologic information on the nature of this disease in northern Alberta. For the 141 female patients who were seen preoperatively in 1971 and thereafter followed regularly, disease was restaged according to the 1973 revision of the tumour-nodes-metastases (TNM) formula of Union internationale contre le cancer (UICC) to permit comparison with staging according to the 1968 formula. The staging distribution was altered greatly because of major changes in the staging system. With the 1973 revision stage II includes disease without palpable lymph nodes and greater emphasis is placed on tumour size. Accurate preoperative staging and registration are essential for meaningful statistics and as an aid in patient management.


Management of the rape victim.

Management of the rape victim. A woman's response to rape can be divided into three phases: an acute reaction, an intermediate stage and a period of resolution. Proper management of the physical and emotional problems of each phase, ideally by the woman's family doctor or gynecologist, may prevent future problems. Treatment during the first phase includes responding to the emotional needs of the patient as well as doing a pelvic and general physical examination to detect any injuries; information for possible legal procedures may be obtained quickly and efficiently. Follow-up particularly psychological, is important in the second and third phases.


Don't forget the children.

Don't forget the children. Serious illness or hospitalization of a parent is potentially destructive to the mental health of the child. Stress is caused by both the separation and the illness itself. If the physician does not ensure that the situation is explained to the child, it is likely the child will be confused and frightened and any attempts at clarification will be blocked in the attendant family dysfunction. The general principles of prevention are to share the facts of the situation with the child and to allow him his natural and varied reactions. The family interview is useful in clearing up misunderstandings, sharing feelings and working out family roles.


Occurrence of two cases of cryptococcosis within 3 months on the same general medical ward. Two cases of systemic cryptococcal infection occurred on the same general medical ward within 3 months. The patients had been on the same ward for 5 days. Both had other diseases associated with depression of cellular immune responses had received immunosuppressive drugs.


Which neuroses need specialist care?

Which neuroses need specialist care? In order to differentiate the neurotic patient who both needs and responds to psychiatric care from the majority of neurotic patients, who do not need this, carefully matched pairs of neurotic patients being treated at psychiatric and nonpsychiatric clinics in Montreal were followed up for 1 year. Improvement was substantial regardless of treatment, and the psychiatrically treated, on the whole, improved only slightly more than the others. However, one type of patient improved greatly under psychiatric care while improving almost not at all without it--introverts who considered themselves unhealthy but found life manageable, had avoided taking time off work or using anxiolytic drugs and appeared to handle their frustrations without repressing their irritation or losing self-control. It is suggested that it is mainly this type of neurotic that should receive specialist referral.


Masculinizing granulosa cell tumour.

Masculinizing granulosa cell tumour. A 23-year-old woman had oligomenorrhea, underdevelopment of the breasts, moderate hirsutism and increased serum testosterone values associated with a benign noncystic granulosa cell tumour of the left ovary. She was frail, irritable and apathetic. Since the age of 7 she had had periodic abdominal pain with nausea, vomiting and dizziness; irritability and occipital headache appeared when she was older. Her symptoms resolved and the masculinization did not progress after the tumour was removed. Only six similar well documented cases have been reported.


Symposium--psychiatric education.

Symposium--psychiatric education. The process of learning in groups, as exemplified by the author's experience in a multidiscipline conference on a surgical ward of a general hospital has been examined. A sequence of six developmental states has been identified: initial ambivalence; passive receptivity; resistance; task orientation; the work group and consolidation. These findings have been compared with those of other studies dealing with sequential stages in group development. The psychiatric consultant's role in expediting these phases and some of the difficulties encountered have been mentioned.


Supervision of psychotherapy with psychiatric residents. 1 model. Training of psychiatric residents should include supervised experience in psychotherapy, whatever the mode of practice the psychiatrist-to-be will later choose. Individual supervision of psychotherapy on patients seen at least once a week for a year offers the best opportunity for learning in this area. The supervisor has a teaching role which includes showing the resident how to look more objectively and insightfully at the material, serving as a partial model of a professional identity, and giving direct didactic help regarding the subject of psychotherapy. Such an experience leads togrowth and development of the resident in his professional role as psychotherapist.


Utilizing reflected countertransference Applying the reflection process as a teaching tool in supervision. There are times when teaching cannot proceed through the abstract presentation of content and must progress by the demonstration of what is to be taught. This is true whenever a student is unable, for whatever reason, to appropriately apply such abstract material to the concrete task before him. This may be due to a number of factors, one of the most common of which is the lack of qualitatively similar experiences to which to refer. The teaching of complicated processes such as psychotherapy is an example, especially when the trainee is relatively unsophisticated and/or seems unable to abstract from discussion of blocking phenomena, where it may become necessary to "act in" the feeling tones which are to be the cues for the behaviour to be learned, instead of persisting with abstract interpretations. When, in supervision, a supervisee recreates the conditions existing in the therapeutic situation, this procedural behaviour is called the "Reflection Process". This process often appears to be unconsciously motivated by the need of the supervisee to solicit from the supervisor a practical demonstration which he does not know how to solicit otherwise. Even if the Reflection Process is due mainly to unresolved, unconscious conflicts of the supervisee, it can be a useful clue as to what transference-countertransference problems he is having difficulties with. The fact that such behaviour on the part of the supervisee may arise out of unresolved unconscious conflictual material should not become a reason for dismissing it. Instead, supervisors should become proficient at recognizing the countertransferential feelings in themselves, which can be an accurate reflection of the feelings troubling the supervisee in his dealings with this patient; and they should recognize the distress signal on the part of the supervisee, which can often be responded to constructively. In the supervision, the supervisor at times can, almost unwittingly, come to play the role which the supervised therapists plays in the supervised therapy and, if not aware of the Reflector Process, he may not realize that the countertransferential feelings which underly his assuming that role may not be primarily a response to the character of the therapist he is supervising but rather to that of the patient. They may also be a reflection of the process problem in the supervised therapy. It is not essential that this process be labeled in the supervision; what is essential is that its ramifications be recognized by the supervisor.


SELF-INDUCED TELEVISION EPILEPSY. A study of 2 patients. A unique method of deliberate self-induction of epileptic seizures, using the TV screen was encountered in two young women. Both suffered from highly photosensitive primary generalized corticoreticular orcentrencephalic epilepsy. Self-precipitation developed in periods of their life when spontaneous attacks no longer occurred. The seizures consisted of absence or generalized tonic clonic attacks. (The latter are almost never encountered in "hand waving", the most common form of self-induction reported in the literature.) The seizures were almost always induced in situations of guilt and/or frustation. They were followed by a feeling of tension relief rather than pleasure. Both patients were reluctant to discuss the circumstances under which self-induction occurred, and had difficulty in accepting psychotherapy or complying with antiepileptic medication. The similarity between the two young women was striking, and suggests that patients with this and other forms of self-induced epilepsy should be more fully studied from a psychiatric point of view.


The oedipus complex. A critical re-examination. The Oedipus complex no longer holds the crucial role in psychodynamics attributed to it by Freud. Attempts to distinguish between oedipal and pre-oedipal disturbances are brought into question, and it is suggested that the complex reflects many non-psychosexual issues, particularly separation-individuation. Ten views of the Oedipus complex are critically reviewed.


Treatment of advanced malignancy with CCNU (NSC 79037): a phase II cooperative study with long-term follow up. CCNU (1-[2-chloroethyl]-3-cyclohexyl-1-nitrosourea, NSC-79037) was used to treat advanced malignancies in 329 evaluable patients. The treatment dosage was 130 mg/m2 for patients with adequate bone marrow reserve and 100 mg/m2 for those with compromised bone marrow. Oral treatment was repeated at 6-week intervals unless hematologic toxicity intervened. There were four complete responses: two in ovarian cancer, one with small cell carcinoma of the lung, and one with melanoma. Tumor response greater than 50% reduction in tumor size occurred in 39 patients (11.9%) while stable disease (no change or decrease or increase of less than 50% in tumor size) was noted in 152 patients (46.2%). Tumor progression occurred in 130 cases. Melanomas and ovarian and lung cancers had the highest response rates. Bone marrow depression was the major side effect of treatment; there was a significant positive correlation between the severity of leukopenia and thrombocytopenia and tumor response to treatment.


Methyl-CCNU, alone and in combination with vincristine, or vincristine and methotrexate, in advanced bronchogenic carcinoma. Ninety-two patients with advanced bronchogenic carcinoma were prospectively randomized according to performance status, histology, and extent of disease to methyl-CCNU alone; methyl-CCNU and vincristine; or methyl-CCNU, vincristine, and methotrexate. Seventy-three patients were evaluable. Randomization to methyl-CCNU alone was discontinued when only three brief "static" responses were noted in 11 patients and the survival (p less than .01) and time on study (p less than .05) were noted to be significantly less than with the combination. Two of 32 patients treated with methyl-CCNU and vincristine, and two of 30 patients treated with methyl-CCNU, vincristine, and methotrexate had mixed responses with a median of 67.5 days.  A static response was seen in eight of 32 and none of 30 patients, respectively. Minimal toxicity occurred in all regimens. Methyl-CCNU alone or in combination with vincristine or vincristine and methotrexate is of limited benefit in patients with lung cancer, although our data suggest (p = 0.15) that the addition of methotrexate increases time on study and survival in patients with extensive disease.


Malignant histiocytosis: complete remission with combination chemotherapy. A case of a 16-year-old girl with malignant histiocytosis involving the lymph nodes, bone marrow, peripheral blood, liver, and spleen is described. This patient achieved a complete remission with cyclic quadruple chemotherapy consisting of cyclophosphamide, vincristine, procarbazine, and prednisone (COPP). She is alive and free of disease more than 4 years after the completion of COPP and after having had maintenance treatment with a small dose of cyclophosphamide for 2 3/4 years. Such good results with the COPP regimen in malignant histiocytosis have not been reported previously, and it is suggested that this modality of treatment warrants further trial.


Skin ulcers due to adriamycin.

Skin ulcers due to adriamycin. Local skin necrosis at the site of intravenous or intra-arterial adriamycin infusion is an infrequent, but serious complication. Ulcers secondary to adriamycin have insidious beginnings, but progress to a much deeper extent than would be expected from their initial appearance. Deep structures, such as tendon or bone, may become exposed. The ulcers are indolent and do not develop a granulation tissue response or epithelialization, as might be expected from their early appearance. Injections of adriamycin in the dorsum of the hand should be avoided when possible, since tendons have little skin cover and the area is difficult to cover with local tissue if there is skin loss. While prevention is important, early surgical treatment may prevent progressive deep involvement and seems warranted when the patient has a life expectancy of months or years. Wide excision of all inflamed tissue is the treatment of choice, with split-thickness skin grafting or flap coverage.


Single view mammography: a simple and efficent approach to breast cancer screening. In 1974 a new approach to breast cancer screening was introduced--the single view mammography method. An urban nonselected population of 6845 women of more than 34 years of age were screened with one roentgenogram of each breast. By this method it has been possible to reduce the radiation dose, while the patient flow is increased and the costs are decreased to an acceptable level. Forty cases of breast carcinoma were detected (6.7/1000). Twenty cases were preclinical.


Routine full-lung tomography in the initial staging and treatment planning of patients with Hodgkin's disease and non-Hodgkin's lymphoma. The yield of additional information from anteroposterior full-lung tomograms that changed stage or treatment, in comparison to that obtained from routine chest radiographs, was prospectively evaluated in 243 previously untreated patients with Hodgkin's disease and non-Hodgkin's lymphoma. Although new information was found in 21.4% of all patients, in only 1.2% did these additional data change patient staging. In 3.3% of the other patients the tomograms provided information that affected radiotherapy treatment planning as practiced in our institution.


The value of the liver scan in preoperative screening of patients with malignancies. This study analyzes the value of the liver scan as a preoperative screening procedure for occult liver metastases in patients with melanoma, sarcoma, head and neck carcinoma, and pelvic carcinoma. The records of 566 consecutive patients admitted to the Surgery Branch of the National Cancer Institute between 1969 and 1974 were reviewed and 323 patients were found acceptable for inclusion in the study. In these patients, although the liver scan had an overall accuracy of 95%, the scan identified only 50% of the patients with occult metastases to the liver and did not siginificantly add to the yield of the other screening procedures. It was useful as an adjuvant to an abnormal routine workup to confirm and localize metastases to the liver. Scans with only non specific abnormalities were of little help. Liver metastases were not identified in any patients with sarcoma, head and neck cancer, or clinically localized carcinoma of the cervix. Therefore, the liver scan was determimed to be an unnecessary part of their screening workup. Patients with recurrent or advanced carcinoma of the cervix and advanced melanoma were found to have an increased incidence of liver metastases. There was a 10% incidence of occult metastases to liver in patients with melanoma and the incidence increased with advancing clinical stage of disease. Even in this high-risk group of patients the screening liver scan did not significantly add information to that gained by history, physical examination, and blood work.


Bone scanning in lymphoma.

Bone scanning in lymphoma. The results of bone scanning with the newer technetium-99m complexes were correlated with clinical, laboratory, and radiographic findings in 26 patients with malignant lymphoma (10 with Hodgkin's disease and 16 with non-Hodgkin's lymphomas). Abnormalities on bone scan compatible with lymphomatous involvement of the skeleton appeared to occur more commonly in patients with diffuse lymphomas than in patients with nodular lymphomas and were generally observed in the setting of advanced disease (15 of 23 patients). Twenty-seven (73%) of the 37 scans obtained were abnormal. Although abnormal scans were observed with the greatest frequency in patients with bone pain (11 of 11), bone marrow involvement (11 of 12), abnormal skeletal radiographs (11 of 11), and elevated serum alkaline phosphatase levels (5 of 6), bone scanning also detected lymphomatous involvement in patients free of pain or with normal laboratory tests. Moreover, conventional radiography was entirely normal in six (35%) of 17 patients with abnormal scans and revealed only nonspecific osteopenia in another two patients (12%). Serial bone scans in nine patients reflected their response to chemotherapy. Of the 37 scans, only one was judged falsely positive and one falsey negative. Bone scanning with 99mTc complexes is a safe, simple, and sensitive screening procedure for detecting both extensive and focal lymphomatous involvement of the skeletal system and is a useful means of following such involvement in response to treatment.


Immunocompetence, immunosuppression, and human breast cancer. I. An analysis of their relationship by known parameters of cell-mediated immunity in well-defined clinical stages of disease. General immune competence was examined 255 breast cancer patients, including 104 operable, 44 locally advanced/inoperable, and 44 with demonstrable metastatic dissemination, all at the time of diagnosis, as well as 63 disease-free long survivors; this was compared with that of 100 normal controls. The parameters employed were PPD and DNCB skin testing, lymphocyte response to PHA mitogen, E-rosette formation, and lymphocyte number. Significant patients, with only 31% showing optimal and 25% showing minimal levels of immune function, as compared with 70% optimal and 2% minimal function in controls. Immune competence was not affected by metastatic involvement of regional lymph nodes. In patients with early, occult metastatic dissemination (as determined in retrospect), the degree of immune competence was found to be identical to that of patients who did not develop disease dissemination. Remarkably, this early phase of tumor spread is not accompanied by immune impairment, such as is evident in clinically demonstrable metastatic disease and, to a lesser degree, in advanced local and regional disease. Since tumor dissemination preceded impairment of general immunocompetence, it emerges as the cause rather than the result of immunosuppression. Long disease-free survivors, who had postoperative irradiation 5-12 years previously, were shown to have a notably low level of immune competence. Lymphocyte response to PHA stimulation was found to be impaired in the earlier stages of disease, while skin DHR was still well maintained; in advanced disease both parameters tend to correlate as total immunologic impairment ensues. The sequence of immunologic events leading up to immunosuppression with disease progress is discussed.


Multiple pulmonary leiomyomatous hamartomas: report of a case with ultrastructure examination. The electron microscopic features of pulmonary leiomyomatous hamartoma, very rare neoplasm, were studied. Characteristic smooth muscle cells containing microfilaments, dense bodies, plaques, pinocytotic vesicles, and basal lamina were present. Fibrosis was minimal, which supports the belief that this component is a secondary phenomena.


On the nature of desmoplastic melanoma.

On the nature of desmoplastic melanoma. Pathologic study demonstrated lentigo maligna melanoma in three of four cases of desmoplastic melanoma and demonstrated spindle cell masses with features of atypical fibroxanthoma in all four of the cases. Pigment enzymes of melanosomes were not found in the mesenchymal portion of the tumor. The cellular atypia and the histochemical and ultrastructural findings favor the fibroblastic nature of this melanoma-related dermal mass.


Siderosis of lymph nodes patients with Hodgkin's disease. Lymph nodes removed from 28 untreated patients with Hodgkin's disease all contained markedly increased amounts of hemosiderine, whether or not they were histologically involved in the disease. This finding was particularly striking in patients with the nodular sclerosis type of disease. Abnormal deposits of iron were also noted frequently in lymph nodes containing metastatic carcinoma, lymphoma of non-Hodgkin's type, and reactive hyperplasia, but in each case, with the exception of metastatic squamous cell carcinoma, the amount was significantly less than seen in Hodgkin's disease. The findings suggest that in patients with Hodgkin's disease and perhaps in those with other disorders in which abnormal tissue retention of iron underlies sideropenic anemia, lymph nodes are an important site of iron retention.


Effects of human and experimental cancer on the conversion of 14C tripalmitin to 14CO2. 14C Tripalmitin was administered intraperitoneally to control mice and to mice bearing subcutaneous Krebs-2 carcinoma. The rate of excretion of 14CO2 was studied. The presence of a tumor induced a greater than 80% suppression of the formation of 14CO2 within 24 hours from the implantation of the tumor, thus allowing unambiguous identification of the tumor-bearing mice. When cancer patients were studied, along with normal volunteers and patients with non-neoplastic diseases, 14CO2 excretion data could be used to identify the cancer patients with an overall accuracy of 84%.


Effects of nitrogen mustard and splenectomy on mouse bone marrow colony formation in vitro. Improved tolerance of splenectomized patients with Hodgkin's disease (HD) to radiotherapy and chemotherapy has been reported. The present study was undertaken to determine the effects of splenectomy and nitrogen mustard (NM) on colony-forming cells (CFC's) of bone marrow cells obtained from CF1 male mice by in bitro agar-gel technique. Splenectomized mice were given NM intraperitoneally on day 11. On day 15, they were sacrificed and the bone marrow was cultured with a source of colony-stimulating factor (CSF). Spleen extract was prepared by grinding spleens from CF1 mice. On the eighth day of incubation, significantly higher numbers of CFC's were found in splenectomized animals at 1% confidence level (F Test) compared with the nonsplenectomized animals. Both splenectomized and non-splenectomized mice had a greater colony response after NM (at 5% confidence level) than saline-treated controls. Maximum numbers of colonies were obtained in the nustard-treated asplenic animals. Splenic extract, as well as extracts from other organs, when added to the culture plates resulted in inhibition of colony formation. The significance of in vitro inhibition after addition of organ extract is uncertain.


Blastic phase of agnogenic myeloid metaplasia simulating malignant lymphoma. A patient with angogenic myeloid metaplasia in blastic transformation with unusually prominent lymphadenopathy simulating lymphoma is described. Interestingly, the initial evidence of myeloblastic transformation was present in the lymph nodes but not in the marrow. Serial bone marrow biopsies showed fibrosis throughout the course of the disease, in spite of a gradual increase in the percentage of peripheral blood myeloblasts. Aneuploidy with a marker chromosome and a Philadelphia-like chromosome was present in the lymph node, bone marrow, and peripheral blood cells. The literature is reviewed regarding the incidence of prominent lymphadenopathy and chromosome abnormalities in agnogenic myeloid metaplasia and other myeloproliferative disorders.


Oncocytic neoplasms of salivary glands: a report of fifteen cases including two malignant oncocytomas. Fifteen oncocytic (oxyphilic granular cell) neoplasms of the salivary glands collected from three large university medical centers are presented. The histologic and ultrastructural criteria that characterize oncocytes are presented and electron micrographs of two of our cases are discussed. Included in the series are 10 benign oncocytomas, two malignant oncocytomas and three benign, oncocytic pleomorphic adenomas. The clinical and pathologic features of benign oncocytomas are reviewed, with special attention to the variety of gross and microscopic changes present. The literature on malignant oncocytomas is critically reviewed, diagnostic criteria are evaluated, and a unique case is discussed in detail. Reference is made to a group of neoplasms that we designate as benign locally aggressive oncocytomas. The problem of differential diagnosis between oncocytoma and bening, oncocytic pleomorphic adenoma is considered.


Carcinoma metastatic to the eye and orbit III. A clinicopathologic study of 28 cases metastatic to the orbit. Two hundred and twenty-seven cases of carcinoma metastatic to the eye and orbit have been reviewed previously. The orbit alone was involved in 28 cases: theses lesions constitute the basis of this clinicopathologic study. The most common signs and symptoms produced by orbital metastasis included exophthalmos (75%1, pain (29%), decreased vision (29%), periorbital swelling (25%), a visible mass (21%), ophthalmoplegia, and diplopia (18%). The ophthalmologists' preoperative (or premortem) clinical diagnoses were: orbital mass, 36%; metastatic carcinoma, 29%; leukemia, 7%; melanosarcoma, 4%; mixed tumor of lacrimal gland, 4%; and meningioma, 4%. The sites of the primary tumors in the 28 patients with orbital metastasis were as follows: breast, eight; lung, four; genitourinary tract, four; pancreas, one; and ileum, one. In 10 patients the site of the primary carcinoma was not determined. In 17 of the 28 patients, symptoms of orbital metastasis preceded detection of a primary tumor elsewhere in the body. In 10 of the remaining 11 patients, detection of the primary tumor had preceded the onset of orbital symptoms. In one patient, symptoms of the primary tumor and of orbital metastasis appeared at about the same time. The median survival of patients with carcinoma metastatic to the orbit was 15.6 months from the time of orbital surgery. This was much better than the median survival of the 227 patients in the overall study (7.4 months) and far better than the median survival of the patients with metastasis to the anterior segment of the eye (only 5.4 months).


Urinary polyamine levels in patients with localized malignancy. Polyamine levels (putrescine, spermidine, and spermine) were determined in 24-hour urine samples by a high voltage electroporesis techique. Twenty-four of 26 patients with localized malignant tumors had two or more elevated urinary polyamine levels. Seven of 12 patients with regional spread of their cancer and five of 11 patients with localized benign and/or noninvasive tumors had elevated urinary polyamine levels. Elevations were seen more frequently frequently in patients with gynecologic tumors. Our data suggest that there is no significant difference between the individual of total polyamine levels obtained in patients with localized malignant tumors, and those levels obtained in patients previously studied with widespread metastatic disease.