Analysis of sera from SV40-immunized and tumor-bearing hosts for blocking activity and antibody-dependent cellular cytotoxicity.

The role of serum factors in tumor immunity to cells transformed by PARA-(defective SV40)-adenovirus 7 was investigated. It was found that sera from SV40-sensitized hosts did not block the specific cytotoxicity of SV40-sensitized spleen cells for PARA-7 cells. However, such sera could collaborate with nonsensitized spleen cells to produce specific killing. This activity could be absorbed out by PARA-7 cells but not by cells transformed by cytomegalovirus. The activity of sera from hamsters bearing tumor isografts depended upon when, after transplantation, the specimens were obtained. Sera collected greater than or equal to 10 days after grafting completely blocked immune spleen cell cytotoxicity and did not mediate target cell killing in the presence of normal spleen cells. Sera obtained at an earlier time, i.e., 3 to 6 days after transplantation, consistently were active in the antibody-dependent cellular cytotoxicity test and exhibited reduced or no blocking of antibody-independent cellular cytotoxicity. Thus, there appears to be an inverse correlation in the capacity of serum from tumor bearing hosts to block effector cell cytotoxicity and mediate antibody-dependent cellular cytotoxicity.


Immunologic effects of neonatal infection with mouse thymic virus.

Mouse thymic virus is a herpesvirus that causes extensive thymic necrosis when given to newborn mice. During the time of acute infection spleen cells have markedly diminished reactivity to T cell phytomitogens and to allogeneic cells and are incapable of effecting a primary in vitro response to a "T-dependent" antigen; responses to B cell mitogens and to a T-independent antigen are unimpaired. Spleens from acutely infected mice have low theta antigen normal numbers of immunoglobulin-bearing cells. Surprisingly, despite widespread necrosis and cellular depletion, thymic cell reactivity to mitogens is unimpaired. However, the ability to thymocytes to proliferate and to generate cytotoxic killer cells in response to allogeneic cells is diminished.


The identification of HL-A antigens on fresh and cultured human endothelial cells.

Human endothelial cells were obtained from the umbilical cord veins of 16 newborns by methods previously described and tested for HL-A antigens by a microcytotoxicity method. HL-A antigens were present on all endothelial cell lines tested. When the HL-A phenotypes of fresh endothelial cells and autologous fetal lymphocytes were compared, a concordance of 70% was observed. When the HL-A phenotypes of maternal lymphocytes and fresh endothelial cells were compared, a maternal contribution to the endothelial cell phenotype was evident in 72% of the possible commmon antigens. Some HL-A antigens were deleted from 11 of 16 endothelial cell lines that were re-typed after 2 weeks in tissue culture. The majority (90%) of deleted antigens were from the second HL-A locus. When three lines of endothelials cells were again re-typed after 6 weeks in culture, no further changes in antigenicity were noted. These findings: a) demonstrate that HL-A antigens are present on human endothelium and suggest that endothelial cells are actively involved in establishing the immunogenicity of a graft, and b) demonstrate that the HL-A antigens on human endothelial cells may be modulated by in vitro culture.


Depression of in vitro responsiveness to phytohemagglutinin in spleen cells cultured from chickens with Marek's disease.

Cultures of dispersed spleen cells, prepared from MDV-infected chickens with MD visceral lymphomas, showed marked depression of responsiveness to the T cell mitogen PHA, as measured by 3H-Tdr incorporation in cells in vitro. When data are expressed quantitatively in terms of cpm/10(5) viable cells, the functional depletion of PHA-responsive cells appear to result from lower levels of 3H-Tdr incorporation in the PHA-stimulated spleen cultures from chickens with acute MD symptoms, as compared to similar cultures from uninfected isolator-reared control chickens. It is suggested that depression of PHA-induced blastogenesis is spleen cell cultures from chickens with acute MD reflects virus-related alterations in T lymphocytes.


Establishment of replicating long-term lines of rabbit macrophages and lymphocytes.

Macrophages (monocyte derived) and lymphocytes have been successfully cultured from rabbit peripheral blood so that replicating long-term cultures are maintained both in media and in the viable, frozen state. The macrophages are considered as such on the basis of phagocytic capabilities, gamma-globulin receptor sites as evidenced by rosette formation with sensitized red cells and disparate enzyme content from the lymphocytes.


An improved quantitative micro-complement fixation test.

The technical procedures for a simple quantitative micro-complement (C) fixation test are described. Major advantages of the present technique compared with the previously described method are: a) a simple measurement of the residual hemolytic activity of C by counting the radioactivity released from 51Cr-labeled sensitized sheep erythrocytes (51Cr-EA); b) an increased sensitivity of the test, brought about by the use of a relatively small number of 51CR-EA per reaction volume; and c) an increased specificity of the test, achieved by maintaining a constant amount of C available for the specific antigen--antibody reaction.


The fine structure of developing human epidermis: light, scanning, and transmission electron microscopy of the periderm.

Eight stages in the development of the human embryonic and fetal periderm have been established, primarily on the basis of surface morphology, major changes in epidermal stratification, and differentiation. The changes in the periderm observed with the scanning electron microscope have been correlated with and supplemented by cytologic studies with the transmission electron microscope in the periderm and other epidermal layers. Light microscopy was used to determine what proportion of the epidermal thickness is accounted for by the periderm and what relationship individual periderm cells have with underlying cells. The results yield a comprehensive, three-dimensional image of the human epidermis during development and support a concept of the periderm as a layer of "dynamic" cells which project superficial blebs, expand in surface area, then regress at the onset of keratinization, leaving only cellular remnants associated with the adult type epidermis.


Cyclic AMP, cyclic GMP, and glucocorticoids as potential metabolic regulators of epidermal proliferation and differentiation.

The two cyclic nucleotides, cyclic AMP and cyclic GMP, appear to be central to the metabolic regulation of cell proliferation and differentiation in various cells. Moreover, in many systems glucocorticoids appear to act in concert with or parallel to cyclic AMP. The available evidence suggests that these three molecular species--cyclic AMP, cyclic GMP, and glucocorticoids--may be essential to the normal regulation of epidermal proliferation and differentiation. In 1970, we suggested that perturbed epidermal homeostasis, exemplified by psoriasis, might be associated with low cellular levels of cyclic AMP and, in 1972, with high levels of cyclic GMP as well. Subsequent measurements of these two cyclic nucleotides in our laboratory showed a probable reduction in the cyclic AMP/cyclic GMP ratio in lesional psoriatic tissue. This led to the hypothesis that the cardinal features of psoriatic epidermis--glycogen accumulation, excessive proliferation, and reduced cell specialization--are the results of this reduced ratio. A corollary of this hypothesis was that a psoriatic lesion could not begin or exist without this altered cyclic nucleotide ratio. Recently, four different agents--lithium, a beta adrenergic blocking agent, antimalarials, and iodide--have been found to exacerbate psoriasis and to reduce the formation of cyclic AMP in various tissues. Consequently we believe that cyclic nucleotides are of central importance in the pathogenesis of the epidermal component of psoriasis.


In vitro activity of silver sulfadiazine against Herpesvirus hominis.

Silver sulfadiazine, at a concentration of 10 mug/ml, inhibits Herpesvirus hominis types 1 and 2 in vitro. The antiviral effect is directly related to concentration of the drug and duration of exposure.


Experimental aminoglycoside nephrotoxicity.

The nephrotoxicities of gentamicin and three other experimental aminoglycosides were compared at a single 60 mg. per kilogram per day dose in rats. Renal function, lysosomal enzymuria, and antibiotic concentrations in plasma, urine, and renal tissue were measured at regular intervals throughout the course of treatment. Kidney tissue was examined by light and electron microscopy in animals killed at intervals throughout the period of antibiotic administration. Proteinuria and enzymuria were early indicators of nephron dysfunction, whereas endogenous creatinine clearance declined later in the course of treatment. All animals were killed 24 hours after a previous antibiotic injection and displayed sustained renal tissue antibiotic concentrations which were 5 to 10 times higher than those in serum or urine. When assayed separately, renal cortical tissue had a fivefold greater antibiotic concentration than renal medulla. Light microscopy displayed necrosis of the pars convoluta of the proximal tubule. Electron microscopy revealed appearance of cytosegrosomes with myeloid bodies. It is possible that impaired cytoplasmic degradation of sequestered organelle membranes, resulting from aminoglycoside accumulation, is responsible for the myeloid body formation and subsequent tubular necrosis.


The spectrum of electrophoretic mobility of very low density lipoproteins: role of slower migrating species in endogenous hypertriglyceridemia (type IV hyperlipoproteinemia) and broad-beta disease (type III).

To test whether beta-migrating very low-density lipoproteins (VLDL, d less than 1.006) might not be unique to broad-beta disease (with a Type III lipoprotein pattern) but rather a caricature of an intermediate species in the catabolism of triglyceride-rich lipoproteins of normal composition and electrophoretic mobility in nonretarding media, VLDL from subjects with endogenous hypertriglyceridemia (with a Type IV pattern) or broad-beta disease were analyzed under varying dietary and pharmacologic conditions following starch-block electrophoresis. These studies revealed a spectrum in electrophoretic mobility and lipid composition throughout the Sf20 to 400 range: the more buoyant, triglyceride-rich VLDL migrated faster and the denser, triglyceride-poor VLDL more slowly, but the VLDL were broadly and continuously distributed throughout the entire beta to alpha2 regions in both disorders. However, in each subfraction of VLDL (Sf100 to 400, 60 to 100 and 20 to 60) as well as in the whole Sf20 to 400 class, the relative proportion of slower species was greater in the subjects with broad-beta disease than in those with endogenous hypertriglyceridemia. Under conditions of acutely stimulated VLDL production (following an oral fat load), a late increase in the slower species was observed as alimentary lipemia resolved. During chronic VLDL hypersecretion (with high carbohydrate feeding) both faster and slower species increased in a subject with broad-beta disease. In the same subject during clofibrate therapy, the faster species were decreased more than the slower on both normal and high carbohydrate diets. Acute acceleration of VLDL catabolism by heparin administration increased the slower VLDL at the expense of the faster, both in this subject and in a counterpart with endogenous hypertriglyceridemia. These studies are consistent with the hypothesis that slower migrating, triglyceride-poor VLDL are normal intermediate (or remnant) forms in a continuous catabolic process. The concentration of these remnants is dwarfed by that of the faster species in subjects with endogenous hypertriglyceridemia. However, in subjects with broad-beta disease they accumulate as the beta-VLDL characteristic of this disorder, most likely as a result of a relative blockade in their further catabolism.


Role of insulin and glucose in the induction of hypoaminoacidemia in man: studies in normal, juvenile diabetic, and insuloma patients.

It is generally assumed, but not established, that the lowering of plasma free amino acid levels following glucose intake is entirely mediated by stimulated insulin secretion. This problem was investigated in groups of healthy volunteer subjects, juvenile diabetics, and insuloma patients. The plasma concentrations of 15 individual free amino acids, glucose, and insulin or C-peptide were determined before and at intervals after either oral glucose (75 Gm.) or intravenous glucose (0.25 Gm. per kilogram of body weight) or tolbutamide (1 Gm.). Glucose lowered plasma amino acid levels in both situations of stimulated (healthy volunteer subjects) and absent (diabetic) insulin secretion. The lack of insulin response in juvenile diabetics was evident by the lack of any significant increase in C-peptide levels following glucose ingestion. Nevertheless, despite markedly greater plasma glucose levels, hypoaminoacidemia was less severe in diabetics than in normal volunteer subjects. In comparison to healthy volunteer subjects who showed significant decreases in concentrations of 14 of 15 examined amino acids by glucose, the diabetic individuals exhibited significant decreases in concentration of 8 amino acids (leucine, isoleucine, valine, tyrosine, phenylalanine, methionine, citrulline, and arginine). In face of a massively greater insulin response in an insuloma patient (greater than 1,000 muU per milliliter), the decreases in the levels of individual free amino acids following glucose were within the range of values obtained in the normal volunteer subjects. The stimulation of insulin secretion with tolbutamide also lowered the concentrations of free amino acids in plasma. However, one importnat difference distinguished the effect of tolbutamide from that of glucose. Within a few minutes after the injection of tolbutamide, there was a marked and persistent fall in alanine levels, while during the entire course of the glucose tolerance test there were no significant changes in the level of this amino acid, suggesting a difference in the mechanism of hypoaminoacidemia induced by endogenous insulin in the presence or absence of exogenous glucose. Despite a markedly greater insulin response to tolbutamide in the insuloma patient (greater than 1,500 muU per milliliter), the decreases in plasma glucose and free amino acid levels fell within the range of values obtained in normal volunteer subjects. In conclusion, the exogenous glucose per se lowers the levels of free amino acids in plasma. This effect is enhanced by insulin.


Renal effects of lithium administration in rats: alterations in water and electrolyte metabolism and the response to vasopressin and cyclic-adenosine monophosphate during prolonged administration.

Lithium (Li+) chloride, 2 to 3 mEq. per kilogram of body weight, was administered intraperitoneally to normal Wistar rats daily for 4 to 66 days. This resulted in a marked reduction in urine osmolality (Uosm.) and increase in the excretion of water, Na+, K+, uric acid, and phosphate. The excretion of uric acid and potassium was a direct function of UNaV. The magnitude of depression in urine osmolality was significantly related to the rate of excretion of lithium in the urine, suggesting that the change in water reabsorption is dependent on the presence of the ion in the luminal side of the tubule. During 2 per cent saline diuresis, Li+-treated rats achieved less fractional free water reabsorption (TcH2O/GFR times 100) at any level of fractional osmolar clearance (Cosm./GFR times 100) than normal rats. On the other hand, during 0.225 per cent saline diuresis, fractional free water clearance (CH2O/GFR times 100) was normal over a wide range of fractional urine flow (V/GFR times 100), indicating intact function of the ascending limb of the loop of Henle. The intravenous infusion of vasopressin (VP) or dibutyryl cyclic-adenosine monophosphate (dcAMP) to Li+-treated rats resulted in a modest rise in Uosm. and a reduction in V/GFR times 100 and CH2O/GFR times 100. Although the response to VP appeared earlier than that to dibutyryl cyclic-AMP, the magnitude of the changes in Uosm., V/GFR times 100, and CH2O/GFR times 100 was eventually the same with both substances. Comparison between normal and Li+-treated rats revealed that the response to both VP and dibutyryl cyclic-AMP was blunted, albeit to a greater extent in the former. Inhibition by Li+ of adenylate cyclase will only partially explain the present data. Impairment in the release of endogenous VP or a block distal to the formation of cyclic-AMP must have played a role. In view of a normal diluting capacity and the increase in the excretion of phosphate and uric acid, it is suggested that Li+, when administered chronically in the present doses, inhibits proximal tubular reabsorption.


Lymphocyte-associated antigens in patients with alveolar cell carcinoma.

Antiserum produced in rabbits against a virus-like particle pellet recovered from human alveolar cell carcinoma (ACC) was used for the detection of lymphocyte-associated antigen in patients with ACC. The method used was direct immunofluorescence. The results demonstrate that lymphocytes from some patients with ACC possess antigen and the presence of lymphocyte fluorescence in patients with ACC suggests widespread or metastatic disease. The presence of antigen may be of both diagnostic and prognostic value.


Measurement of unsaturated ""granulocyte-related'' (TC I and TC III) and ""liver-related'' (TC II) B12 binders by instant batch separation using a microfine precipitate of silica (QUSO G32).

A microfine precipitate of silica (Quso G32) almost instantly quantitatively adsorbs the polypeptide transcobalamin II from serum without adsorbing the glycoproteins transcobalamins I and III. This provides a simple, rapid, and reproducible method for the measurement of the granulocyte-related transcobalamins I and III and the liver-related transcobalamin II in serum, and is also applicable to their purification, and the separation and purification of other polypeptide-glycoprotein mixtures.


Extranasopharyngeal juvenile angiofibroma.

A rare case of juvenile angiofibroma arising from the base of the skull outside the nasopharynx and extending into the pterygo-maxillary fossa and cheek is discribed. It is possible, by a  combined zygomatic and buccal approach, successfully to excise such a tumour.


Uptake of 125-I-labelled human placental lactogen and human placental lactogen by the tissues of normal and lactating rats.

The rate of clearance from the circulation and uptake into tissues of radioactive label was studied after i.v. injection of 125-I-labelled human placental lactogen (HPL) into rats at various stages of pregnancy. The half-life was obtained for the disappearance of the trichloroacetic acid-precipitable material from the plasma. The half-life, t1/2(S), calculated over the first 5 min after injection of the hormone was 5.4 equals or minus 1.1 (S.D.) min, while a half-life, t1/2(L), of 27.9 equals or minus 2.3 min was obtained from the decay period of 15-35 min. In the non-pregnant and pregnant rat the highest ratio of the radioactivity in an organ to that in the blood was 12-14:1 in the kidney. That the kidney is mainly involved in the uptake of exogenous HPL is further confirmed by the application of the histochemical immunoperoxidase technique. Human placental lactogen was localized in the cells of the proximal tubules of the cortex and to a lesser extent in the tubular lumen and the tubules of the medulla region. UPTAKE OF HPL in vivo occurs in the mammary gland tissue of the post-partum rat and reaches a maximum uptake between 15 and 30 min after injection of the hormone. Furthermore, specific uptake of HPL was observed on the alveolar cell membranes after the incubation of paraffin-embedded sections of formalin-fixed mammary gland and subsequent treatment by the peroxidase-labelled antibody method. These findings support the work of others who have demonstrated the presence of specific membrane receptors in the mammary gland for hormones with prolactin-like activity.


Oestradiol-induced synthesis of a specific uterine protein in propranolol-treated rats.

While there is conflicting evidence concerning an effect of oestradiol on uterine cyclic AMP concentration, results from different laboratories (including ours) are in agreement that even when observed, the early increase in uterine cyclic AMP after oestradiol injection fails to occur when propranolol, a beta-adrenergic blocking agent, is given (50 mug, i.p.) 20 min before the oestradiol. The present work shows that pretreatment with propranolol failed to inhibit an early uterine response to oestradiol, namely the synthesis after 1 h of uterine protein, or class of proteins, IP. It is concluded that the induction of IP by oestradiol does not depend on an increase in uterine cyclic AMP concentration and that beta-adrenergic receptors do not have a role in this oestrogenic response.


Synthesis and secretion of a large glycoprotein in the pars intermedia.

The biosynthesis, intracellular transport and release of [3-H]-leucine-containing secretory product has been followed in the pars intermedia of Xenopus laevis and, in particular, the synthesis and secretion of a large molecular weight glycoprotein secretory product was demonstrated. However, if Xenopus adrenocorticotrophin does contain a leucine residue the results obtained provided no support for the view that it serves as a precursor for melanocytestimulating hormone in this species.


Aspects of the adrenal function in the domestic fowl.

Chromatographic purification of extracts of hen peripheral plasma on Florisil columns before measurement by spectrofluorometry showed a basal level for corticosterone of 1-3 mug/100 ml, which is much lower than concentrations previously reported using acid fluorescence. Neither handling, restraint nor repeated bleeding affected the concentration of this hormone or of glucose. Adrenal function tests with two preparations of synthetic corticotrophin showed that they caused a rapid rise in blood corticosterone and eventually of glucose. Results of studies using insulin or tolbutamide i.v. suggest that there is a threshold concentration of plasma glucose (about 70 mg/100 ml) below which hypoglycaemia stimulates the hypothalamic-pituitary-adrenal system in fowls. Prolonged treatment of laying hens with protamine zinc insulin led to aphagia and cessation of egg-laying; increased concentrations of corticosterone were observed 2 days after the administration of insulin ceased, coinciding with the return to normal plasma levels of glucose. 'Chemical adrenalectomy' with metyrapone showed that the restoration of plasma glucose to a normal concentration after insulin treatment is dependent upon fully functional adrenal cortical tissue. It appears likely that the adrenal medulla is a target for corticosterone which probably regulates the tissue levels of phenylethanolamine-N-methyltransferase, one of the enzymes necessary for the biosynthesis of adrenaline.


A radioimmunoassay for plasma arginine-vasopressin in man and dog: application to physiological and pathological states.

A radioimmunoassay has been developed for plasma arginine-vasopressin in man and dog. The mean recovery of added arginine-vasopressin (AVP) was 60 +/-6.9 (S.D.)% and the lower threshold of detection 2.0 pmol/1. A close correlation was found between concurrent radioimmunoassay and bioassay values. The mean concentration found in peripheral venous blood in healthy men after overnight fasting was 5.3 pmol/1 (range 4.6-6.2 pmol/1.) In man, significant increases in plasma AVP occurred after dehydration (5-9-9-5 pmol/1) and significant decreases after oral water-loading (5.9-9.5 pmol/1). During i.v. infusion of graded doses of synthetic AVP in normal men, plasma levels were closely correlated with infusion rate. On stopping the infusion, plasma vasopressin fell exponentially with a half-life of between 7 and 8 min. In man, plasma AVP was unaffected by tilting head-up for 2 h, or by a non-hypotensive bleeding of 500 ml in 10 min. In the dog, haemorrhage of 5 ml/kg and over caused proportionate increases in AVP in the circulation. In normal men, plasma vasopressin was significantly correlated with concurrent urinary osmolality. Five patients with oat-cell bronchial carcinoma and hyponatraemia showed a marked increase of plasma vasopressin. Five patients with diabetes insipidus  had significantly reduced, but detectable, levels of plasma AVP. The plasma concentration in these patients did not increase after water restriction.


The tip of the Dictyostelium discoideum pseudoplasmodium as an organizer.

We have extended Raper's original work on the organizing ability of the tip of the Dictyostelium discoideum slug. Our new results are that tips from all multicellular (pseudoplasmodial) stages act as organizers; that the structure organized by a tip depends on the developmental stage of the cells responding to the tip's signal; that tips from all stages release a qualitatively similar signal which is continuous, most probably a gradient of c-AMP; and that the signal from fruiting-body tips appears stronger than that from conus tips. We discuss these results with reference to the control of morphogenetic movement and patterned differentiation and point out that the D. discoideum tip is analagous to a classical organizer, whose signal is interpreted according to the state of determination of cells in its field of influence.


Monosynaptic connexions between wing stretch receptors and flight motoneurones of the locust.

1. The connexions between stretch receptors of the wings and motoneurones innervating flight muscles have been studied anatomically and physiologically. 2. Filling with cobaltous chloride shows that the single neurone of a forewing stretch receptor has a complex pattern of branches within the mesothoracic ganglion and branches which extend into the pro- and meta-thoracic ganglia. The single neurone of a hindwing stretch receptor has extensive branches in the metathoracic ganglion and branches in themesothoracic ganglion. The branches of both receptors are confined to the ipsilateral halves of the ganglia. 3. A stretch receptor gives information about the velocity and extent of elevation of a wing. 4. Each spike of a forewing stretch receptor casuses an EPSP in ipsilateral mesothoracic depressor motoneurones and an IPSP in elevators. The connexions are thought to be monosynaptic for the following reasons. The EPSPs in the first basalar (depressor) motoneurone follow each spike of the stretch receptor at a frequency of 125 Hz and with a constant latency of about 1 msec. In a Ringer solution containing 20 mM-Mg2+ the amplitude EPSP declines gradually. The IPSP'S upon elevators have similar properties but occur with a latency of 4-6 msec. 5. The connexions therefore comprise a monosynaptic negative feed-back loop; elevation of the wing excites the stretch receptor which then inhibits the elevator motoneurones and excites the depressors. 6. A hindwing stretch receptor synapses upon metathoracic flight motoneurones in the same way, causing EPSPs  in depressor and IPSPs in elevator motoneurones. 7. No connexions of either fore- or hindwing stretch receptors have been found with contralateral flight motoneurones. 8. Interganglionic connexions are made by both receptors. For example, both fore- and hindwing stretch receptors cause EPSPs upon the meso- and metathoracic first basalar motoneurones. 9. Stimulation of the axon of a stretch receptor with groups of three stimuli repeated every 50-100 msec thus simulating the pattern which it shows during flight, causes subthreshold waves of depolarization in depressor motoneurones. When summed with an unpatterned input, the stretch receptor is able to influence the production of spikes in motoneurones on each cycle. During flight, it is expected that the stretch receptor will influence the time at which a motoneurone will spike and hence have an effect on the amplitude of the upstroke and upon the phase relationship between spikes of motoneurones.


Proprioceptive motor control in fish respiration.

The response of single respiratory neurones in the medulla oblongata of carp to short twitches of individual respiratory muscles were analysed. The muscle contractions were obtained through automatic electrical stimulation and could be consistently elicited in a predetermined phase relation to the ventilatory cycle. The results show that, apart from nerve cells which take part in long-term processing of proprioceptive information from several sources, neurones also exist which possess the properties of elements of a peripheral proprioceptive control loop such as tension receptor neurones, length or stretch receptor neurones and motor neurones.


Influence of nitrate on fermentation pattern, molar growth yields and synthesis of cytochrome b in Propionibacterium pentosaceum.

Under anaerobic conditions, Propionibacterium pentosaceum reduces nitrate to nitrite until nitrate is exhausted from the medium when nitrite is converted into N2 or N2O. In the presence of nitrate, fermentation patterns for lactate, glycerol and pyruvate were different from those obtained during anaerobic growth without an inorganic electron acceptor. In the presence of these substrates, a drastic decrease in propionate formation was observed, some pyruvate accumulated during growth with lactate, and acetate was produced from glycerol. Acetate production from lactate and pyruvate was not influenced by the presence of nitrate. Furthermore, CO2 was produced by citric acid cycle activity. The fermentation pattern during nitrite reduction resembled that of P. pentosaceum grown anaerobically without an inorganic electron acceptor. Nitrits has a toxic effect, since bacteria inoculated into a medium with 9 mM-nitrite failed to grow. The cytochrome spectrum of anaerobically grown P. pentosaceum was similar with and without nitrate. In membrane fractions of bacteria grown anaerobically with nitrate, cytochrome b functioned in the transfer of electrons from lactate, glycerol I-phosphate and NADH to nitrate. Molar growth yeilds were increased in the presence of nitrate, indicating an increased production of ATP. This could be explained by citric acid cycle activity, and by ocidative phosphorylation coupled to nitrate reduction. Assuming that I mol ATP is formed in the electron transfer from lactate or glycerol I-phosphate to nitrate, and that 2 mol ATP are formed in the electron transfer from NADH to nitrate, YATP values (g dry wt bacteria/mol ATP) were obtained of between 5-0 and 12-6. The higher YATP values were similar to those obtained during anaerobic growth without an inorganic electron acceptor. This supports the assumptions about the efficiency of oxidative phosphorylation for electron transport to nitrate. Low YAPT values were found when high concentrations of nitrite (15 to 50 mM) accumulated, and were probably due to the toxic effect of nitrite.


Purification and characterization of a polyphosphate kinase from Arthrobacter atrocyaneus.

Polyphosphate kinase, an enzyme which incorporated the gamma-phosphate of ATP into long-chain polyphosphate molecules, was purified more than 700-fold from Arthrobacter atrocyaneus by ammonium sulphate fractionation, DEAE-cellulose column chromatography and Ssphadex G-200 gel filtration. The enzyme had a broad pH optimum at 6-0 to 7-0 and required Mn2+ or Mg2+, histone, and inorganic phosphate for activity. The Km for Mn-ATP was 0-53 mM, and for inorganic phosphate was 1-67 mM. Free ATP concentrations greater than 8 muM inhibited the enzyme. Free Mn2+ or Mg2+ concentrations greater than 2 mM or 6 mM, respectively, were also inhibitory. Activity was strongly inhibited by 4 mM-ADP, 1 mM-PP1 or 20 mM-NaF. The effect of ADP might have resulted from reversing the equilibrium of the kinase reaction. The activation by phosphate ions might indicate a role for the enzyme in regulating intracellular phosphate levels or maintaining a phosphorus reserve. The level of enzymic activity in the bacteria responded to changes in inorganic phosphate concentration in the medium. Basic proteins, such as protamine, could substitute for histone as activator. Proteins such as casein or bovine serum albunim would also substitute for histone but only in the absence of inorganic phosphate. The presence of a protein might be necessary to form a complex with the product, thus preventing reversal of the reaction in vitro. The reaction product was characterized, and found to be labile in hydroxylamine, base, and acid at 100 degrees C. It behaved as a long-chain-polyphosphate molecule on chromatography in an Ebel's solvent. The enzymic activity was therefore not that of a protein kinase.


Taxonomy of the Clostridia: ribosomal ribonucleic acid homologies among the species.

rRNA homologies have been determined on reference strains representing 56 species of Clostridium. Competition experiments using tritium-labelled 23S rRNA were employed. The majority of the species had DNA with 27 to 28% guanine plus cytosine (%GC). These fell into rRNA homology groups I and II, which were well defined, and a third group which consisted of species which did not belong in groups I and II. Species whose DNA was 41 to 45% GC comprised a fourth group. Thirty species were placed into rRNA homology group I on the basis of having 50% or greater homology with Clostridium butyricum, C. perfringens, C. carnis, C. sporogenes, C. novyi or C. pasteurianum. Ten subgroups were delineated in homology group I. Species in each subgroup either had high homology with a particular reference species or a similar pattern of homologies to all of the reference organisms. The eleven species in rRNA homology group II had 69% or greater homology to C. lituseburense. Species in groups I and II had intergroup homologies of 20 to 40%. The six species in group II had very low homologies with groups I and II. Negligible homology also resulted when five of the species were tested against the sixth, C. ramosum. The five species having DNA with 41 to 45% GC were C. innocuum, C. sphenoides, C. indolis, C. barkeri and C. orotic um. Little rRNA homology was apparent between C. innocuum and the other high % GC species or with several Bacillus species having similar %GC DNA. Correlations between homology results and phenotypic characteristics are discussed.


Temporal genetic mapping in the blue-green alga Anacystis nidulans using ethyl methanesulphonate.

Cultures of the blue-green alga Anacytis nidulans were synchronized with respect to DNA synthesis as well as cell division. Application of ethyl methanesulphonate at different stages of replication resulted in a peak of mutation frequency for different genetic markers; this peak can be accounted for in terms of the involvement of repair processes. A temporal map of 19 markers has been constructed by this method. Comparison of gene position obtained by temporal mapping indicates that either bidirectional replication or unidirectional replication from more than one origin occurs.


The action of Vibrio cholerae and Corynebacterium diphtheriae neuraminidases on the Sendai virus receptor of human erythrocytes.

Studies of the Sendai virus haemagglutinin receptor on the human erythrocyte surface have confirmed that it involves 2 leads to 3 linked sialic acid. Because the primary specificity of Vibrio cholerae neuraminidase is for this linkage, it is able to compete with the virus for the receptor, to which it adsorbs strongly at low temperatures. Corynebacterium diphtheriae neuraminidase, whose principal specificity is for a sialic acid linkage other than 2 leads to 3, does not easily remove Sendai virus receptors, nor does it adsorb to the erythrocyte surface. A new definition of the term "receptor-destroying enzyme" is given which takes both enzyme and virus specificity into account, and a modified assay method is suggested in order to overcome the problems due to enzyme adsorption.


Individual differences in compensatory rebound of REM sleep, with particular reference to their relationship to personality and behavioral characteristics.

In subjecting 14 healthy university students to partial differential rapid eye movement (REM) sleep deprivation (PDRD), the compensatory rebound of REM sleep during the next night was determined, and showed fairly substantial individual differences in the increased percentage of REM sleep time. This rate was approximately the same for the same individual for two sleep recordings. These individual differences had no positive correlation with the decreased rate of REM time in the PDRD nights or with the percentage of REM time in the baseline night. Therefore, the individual differences in the increased percentage of REM time can be presumed to reflect individual differences in need for deprived REM sleep. Next, we looked into the relationship between the individual subject's personality and behavior characteristics, and his increased percentage of REM time. This revealed that the individuals who were extrovertive, active, optimistic, showy, and who had many friends had significantly higher increases in the percentage of REM time than the individuals who were introvertive, neurotic, inactive, nervous, modest, and who had few friends. Also discussed was the neurophysiological and biochemical basis of the central nervous system as the background for the relationship between the personality and behavioral characteristics and the increased percentage of REM sleep time.


The terminal autonomic nervous system in the pancreas of newts. Electron microscopic investigations using acetylcholinesterase and zinc iodide-osmium tetroxide reactions.

The intrinsic innervation in the pancreas of newts Triturus vulgaris, Tr. montandoni and Notophthalmus. All the investigated species display: cholinergic innervation better developed than adrenergic, no qualitative differences regarding the fine structure of nerve fibers and terminals as compared to that previously described in the pancreas or in other glands, axo-axonal contacts. Their functional signification as peripheric synapses is suggested. Frequently neuro-glandular junctions with or without membrane specializations. The occurrence of the latter is discussed in connection with peripheral transmission of nervous impulse.


Peripheral nerve involvement in Batten-Spielmeyer-Vogt's disease.

Electromyography and sensory and motor nerve conduction were studied in 23 patients with Batten-Spielmeyer-Vogt's disease ("juvenile amaurotic idiocy", cerebral ceroidlipofuscinosis). A slight to moderate slowing of the sensory conduction velocity was found in the median as well as in the sural nerve, more pronounced in the distal than in the proximal segments. The findings are interpreted as evidence of impaired transmission of the peripheral nerves in Batten-Spielmeyer-Vogt's disease.


Progressive multifocal leucoencephalopathy: remission with cytarabine.

A patient with a 14 year history of sarcoidosis developed a progressive left cerebral hemisphere lesion. The clinical diagnosis of progressive multifocal leucoencephalopathy was confirmed by brain biopsy and remission occurred after treatment with cytosine arabinoside.


Myxoedematous polyneuropathy: a light and electron microscopic study of the peripheral nerve and muscle.

Histopathological findings of biopsied peripheral nerve and muscle were studied in a case with myxoedematous polyneuropathy. The most striking findings in the sural nerve were segmental demyelination and onion bulb formation with scanty mucinous deposits in addition to marked loss of large myelinated nerve fibres. The peroneus brevis muscle revealed the association of neuropathic and myopathic changes. It is suggested that myxoedematous polyneuropathy might be intrinsic neuropathy due to metabolic disorder of Schwann cells related to hypothyroidism, resulting in segmental demyelination, not merely compressive neuropathy due to mucinous deposits in the peripheral nerves.


Peripheral neuropathy in acrodermatitis chronica atrophicans (Herxheimer).

Acrodermatitis chronica atrophicans is a dermatological condition that takes a chronically progressive course and finally leads to a widespread atrophy of the skin. Involvement of the peripheral nervous system is frequently observed, predominantly a sensory polyneuropathy. General reactions, the effect of penicillin treatment, the histological findings, and reports concerning a communicable agent transmittable from human to human as well in tissue cultures point to an infectious disease. Acrodermatitis chronica atrophicans follows a peculiar geographical distribution forming clusters of high prevalence in certain regions. Transmission by ticks is suggested.


A case of type 1 muscle fibre hypotrophy and internal nuclei.

A 14 year old boy was diagnosed as suffering from type 1 muscle fibre hypotrophy with internal nuclei. On histological examination of a biopsied muscle, there was selective hypotrophy of type 1 muscle fibre with internal nuclei, and focal degenerative changes were seen in a few type 1 fibres. The small type 1 fibres were arranged in small or large groups in one bundle. An EMG study of moderately weak muscles revealed low amplitude and short duration motor unit potentials as well as normal potentials and no spontaneous discharges. The H reflexes were abnormally low in amplitude comapred with the M response. The histological and electrophysiological findings suggested that the type 1 fibre involvement in the present case may have a neurogenic basis. It is likely that the clinical features of the reported cases are too variable for a single clinical entity.


Tuberous sclerosis and Klippel-Trenaunay-Weber syndromes. Association of two complete phakomatoses in a single individual.

Tuberous sclerosis and Klippel-Trenaunay-Weber (KTW) syndromes are phakomatoses which are believed to be inherited separately. A 41 year old woman presented with the classic features of tuberous sclerosis: adenoma sebaceum, mental retardation, and seizures. In addition, the diagnostic triad of KTW involved the left lower limb: cutaneous naevi, a vascular anomaly, and osteohypertrophy. Arteriography documented the presence of visceral tumours and an arteriovenous malformation of the leg. This is the first reported association of the fully-developed symptomatology of tuberous sclerosis and KTW in one person.


Myasthenic decrement and myasthenic myopathy. A study on the effects of thymectomy.

Motor unit action potentials, M responses to repetitive nerve stimulation, and anticholinesterase tests were investigated in 12 myasthenic patients before and after thymectomy. In six of them the endarterial acetylcholine test was also carried out. Responsiveness to ACTH or to prednisone treatment was evaluated before and after thymectomy. The typical myasthenic presynaptic disorders were improved by thymectomy, while signs of myasthenic myopathy (according to Rowland's definition) were apparently increased. This process of 'functional myopathophanerosis' is discussed and explained in terms of a previous presynaptic disorder blocking the voluntary recruitment threshold of those motor units which are most affected at both presynaptic and postsynaptic level.


Acrylamide encephaloneuropathy due to well water pollution.

All five members of a family developed subacutely mental confusion and/or truncal ataxia. Symptoms and signs of polyneuropathy were seen later. The well water in the patients' home contained 400 ppm acrylamide. The present cases are unique in that they are cases of acrylamide poisoning induced by oral intake and percutaneous penetration, and that central nervous system symptoms were prominent.


[Atypical juvenile neurolipidosis. Ultrastructural study of a cerebral biopsy].

Electron-microscopic findings on a brain biopsy in a 9-year-old male affected by juvenile lipidosis are reported. Two types of neuronal cytosomes were discribed: lamellar concentric bodies, resembling membranous cytoplasmic bodies, and polymorphous complex bodies, unit membrane bound including various material, mostly resembling lipofuscin. The same storage was found in glial and endothelial cells. Neurochemical data failed to pin-point any specific abnormalities. The present case cannot be classified in the category of neuronal ceroid-lipofuscinosis but seems similar to certain reported cases of atypicalneurolipidosis.


Therapeutic percutaneous embolization for extra-axial vascular lesions of the head, neck, and spine.

Therapeutic percutaneous embolization of extra-axial vascular tumors and arteriovenous malformations was performed 41 times in 27 patients. Twenty-one patients (78%) had a clinically favorable result. In 11 of these patients, the procedure was preoperative and caused a dramatic reduction of surgical blood loss. In the remaining 10 patients with a favorable result, therapeutic embolization alone resulted in a significant clinical amelioration documented by a detailed follow-up varying from 2 to 5 years. In patients with uncontrollable epistaxis, the procedure was life-saving. The guidelines and instrumentation for a safe and effective technique are presented, based on the authors' experience with more than 100 cases of vascular lesions of the brain and spinal cord. A low-viscosity silicone polymer was developed by the authors and used clinically as an intravascular adhesive for the embolization of vascular tumors.


Critical evaluation of hepatic scintiangiography for neoplastic tumors of the liver.

Dynamic hepatic scintiangiography increases the specificity of diagnosis of space-occupying lesions of the liver seen on hepatic scintigraphy. The purpose of this study was to evaluate and compare critically this procedure with histologic and radiopaque diagnosis in the evaluation of suspected hepatic neoplasms. Ninety-two patients had hepatic scintiangiography, scintigraphy, and histologic certification. In ten of these patients the findings of radiopaque arteriography were compared with those of hepatic scintiangiography. In all ten patients with hepatoma, the scintiangiographic and histologic observations correlated; nine of these ten patients had a "tumor stain." Fifty-one of 59 patients with metastases to the liver had scinitangiograms that showed "tumor stain." In 2 of these 59 patients, scintiangiography revealed tumor vascularity whereas the results of scintigraphy were normal. In two of four patients with metastases and two of six patients with hepatomas, scintiangiograms revealed "tumor stain" that was not evident on radiopaque afteriography. Conclusions from this study are: (A) neoplastic arterialization or "tumor stain" is more readily detected by scintiangiography than by radiopaque arteriography; (B) a normal scintigram and a "tumor stain" on the scintiangiogram in a patient with a known primary neoplasm outside the liver is suggestive of hepatic metastases; and (C) a normal scintigram and scintiangiogram make neoplastic involvement of the liver improbable. Dynamic hepatic scintiangiography is a simple, clinically useful method for increasing the specificity of diagnosis of diagnosis of space-occupying lesions of the liver and should be part of the evaluation for possible neoplastic involvement.


The value of hepatic scintiangiography and static liver scans in superior vena caval obstruction: Case report.

Isolated increased uptake of 99mTc-sulfur colloid in a static liver scan with associated hepatic scintiangiography demonstration of collateral venous circulation is pathognomonic for superior vena caval obstruction. A case with these findings is reported and the differential diagnosis of "hot spots" occurring in liver scanning is discussed.


Evaluation of a rapid and simple technique for the radioimmunoassay of triiodothyronine (T3).

This report compares T3 measurements obtained on the same serum samples by a resinstrip technique and by another existing radioimmunoassay method. The samples analyzed were obtained from a total of 90 subjects who were clinically categorized as hypothyroid, normal, hyperthyroid, or euthyroid while taking estrogen-containing compounds or while pregnant. The correlation coefficient for all 90 sera with these two different techniques was 0.94. All subjects who were clinically euthyroid (32) had a normal serum T3 concentration by the resin-strip technique. Similarly, 23 clinically hyperthyroid patients had elevated serum T3 concentrations and 15 of 17 clinically hypothyroid patients had decreased serum T3 levels.


Enzymic formation of thiamine pyrophosphate in plants.

Evidence was presented by paper chromatographic analysis on the occurrence of an enzyme capable of catalyzing a pyrophosphate transfer from ATP to thiamine in green leaves of various plants. The exclusive localization of the enzyme activity in the 105,000 X g supernatant (in a soluble form) was demonstrated by differential centrifugation of a cell homogentae in 0.25 M sucrose. The enzyme was purified by column chromatography with DEAE-cellulose and by gel filtration with Sephadex G-150. The partially pruified preparation, while contaminated with detectable activity of acid phosphatase, lost the ability of utilizing thiamine monophosphate as the substrate in place of thiamine. These findings lead to the conclusion that thiamine pyrophosphate is formed in green leaves of plants through a direct pyrophosphorylation of thiamine in the presence of ATP and Mg.


Cystic partially differentiated nephroblastoma.

An unusual cystic renal tumour is described which resembles, superficially, the solitary multilocular cyst of the kidney, but which shows appreciable amounts of proliferating undifferentiated renal blastema. We believe that this tumour represents a partially differentiated nephroblastoma and that, as such, it provides support for the theory that Wilms' tumour is capable of undergoing maturation to a fully differentiated, benign cystic lesion.


Macrophages in brain tumours induced transplacentally by N-ethyl-N-nitrosourea in rats: an electron-microscope study.

The fine structure of macrophages has been studied in experimental brain tumours induced transplacentally in BD-IX rats by a single intravenous injection of 30 mg of N-ethyl-N-nitrosourea per kg of body weight on the 15th day of gestation. The tumours, depending on their localisation and size, caused various lesions in the brain, namely axonal degeneration, loss of myelin, oedema, haemorrhage and cell necrosis. The tumours and the resulting alterations elicited a strong reaction by macrophages: activation of microglial cells in situ and infiltration of the brain by leucocytes, chiefly by monocytes. Since both microglial cells and monocytes underwent morphological changes, it was difficult, or impossible, to establish the origin of these reacting cells. In a few cases, however, microglial cells and monocytes could be distinguished; this indicated that microglial cells were still being activated and leucocytes were still entering the brain. Various stages of activity of macrophages have been described: the number of lysosomes and cytoplasmic inclusions were thought to indicate activation, phagocytosis and repletion. Activation is characterised by an increase of lysosomes and frequent cell divisions. Phagocytic activity is accompanied by the appearance of inclusions which varied in different lesions: protein-like material in oedema, remnants of erythrocytes in haemorrhages and myelin-lamellae with lipid droplets in demyelination. These various inclusions were frequently present in the same cell, since the different lesions not uncommonly occurred at the same time. In the stage of repletion macrophages contained mainly lipid droplets and unidentifiable debris in their abundant cytoplasm and thus corresponded to the compound granular corpuscles.


Further studies on carrageenan-induced pleurisy in rats.

A study has been made of the exudate provoked by intrapleural carrageenan in rats. Various parameters have been measured including volume of exudate, numbers and types of cells. Pharmacological assay has shown the early release of histamine and 5HT into the exudate followed at later times by a release of prostaglandins, and a second release of histamine. The role of complement has been studied. Intrapleural injection of carrageenan after complement depletion produced a reduced migration of polymorphonuclear cells and also a reduction in volume of exudate. Prostaglandins remained unaffected by this treatment.


Structural and functional characteristics of the microcirculation in neoplasms.

The microcirculation in a transplantable rat fibrosarcoma was investigated. The vasculature of the neoplasm consisted of irregular channels lined by plump endothelium which displayed mainly pentalaminar although a few heptalaminar junctions were also found. Few pericytes surround the endothelial layer while the basement-membrane varied in thickness and was often duplicated and triplicated. Challenge with histamine resulted in increased permeability in comparison with normal connective tissues similarly treated. The increased permeability was accompanied by the formation of interendothelial gaps in these irregular vascular channels. Carrageenan induced a leucocytic exudate in the neoplasm which, apart from an increase in the mononuclear concentration 2 wk after injection, varied little from normal connective tissues. It is concluded that the majority of vascular channels in this neoplasm bear some structural and functional relationship to small venules.


Idiopathic thrombocytopenic purpura in children.

Idiopathic thrombocytopenic purpura in children 10 years of age or younger was observed to have a more favorable prognosis than in older children. Corticosteroid therapy in children judged to be at increased risk of serious hemorrhage resulted in a significantly greater number of patients with an early increase in platelets than was noted in a control group. All patients with chronic disease who responded to administration of a corticosteroid initially and then relapsed had some response to a subsequent course of therapy, although none had a sustained remission. In such patients, splenectomy was a more effective therapeutic measure than treatment with either a corticosteroid or a cytotoxic agent.


Pseudohermaphroditis, with testes and a 46, XX karyotype.

A 14 4/12-year-old white girl, evaluated for progressive virilization and clitormegaly, was found to have the unusual combination of a 46, XX karyotype, well-developed Mullerian structures, and dysgenetic testes with Leydig cell hyperplasia. Although there have been previous case reports of 46, XX males, in all of these patients development of the Mullerian ducts had been suppressed. When contemporary classifications of human disorders of sexual differentation were reviewed, no report of a similar patient was found. We speculate that the genotype and phenotype in our patient correspond to the genetic intersexuality of the hornless goat, thereby raising the possibility that the human autosome may play a role in the control of sexual development.


Angiotensin-I-converting enzyme activity in idiopathic respiratory distress syndrome of the newborn infant and in experimental alveolar hypoxia in mice.

Serum angiotensin-I-converting enzyme activity was found to be elevated in infants with idiopathic respiratory distress syndrome when compared with healthy premature infants, normal infants, and acutely ill full-term infants. Serum and lung CE activity has been found to be elevated in mice exposed to hypobaric alveolar hypoxia which also stimulated renal renin production. These findings suggest that alveolar hypoxia stimulates the renin-angiotensin-aldosterone system and this system may be involved in the response to the stress of IRDS.


Cytomegaloviremia in children with acute lymphocytic leukemia.

Leukocyte and urine cultures were done at monthly intervals in 36 children with acute lymphocytic leukemia known to be excreting cytomegalovirus in their or saliva in order to determine the relationship of viremia to clinical cytomegalic inclusion disease. Eleven of 36 (30.5%) patients had viremia. Viremia was related to clinical disease in only three patients; two with chorioretinitis and one with a CMV monomucleosis syndrom. However, the presence of viremia did not serve as a useful means to determine active CID. Viremic patients with CID all had elevated serum levels of IgM and multiple episodes of viremia. Viremia was not related to the duration, type or number of drugs used in immunosuppression, nor to the hematologic status of leukemia. Viremic patients received more blood transfusions than noviremic patients, but the administration of blood products could not be related to the acquisition of infection. Leukopenia, neutropenia, total lymphocyte count, fourfold rise or fall in complement-fixing titer, and viruria had no consistent relationship to viremia or clinical CID.


Prenatal diagnosis and fetal pathology of I-cell disease (mucolipidosis type II).

Increased activity of several lysosomal hydrolases was demonstrated in amniotic fluid from a fifteenth week pregnancy in which the fetus had I-cell disease. Cultured cells from amniotic fluid had a decreased activity of the same enzymes. The diagnosis of I-cell disease was later confirmed by enzyme assays in cell cultures of fetal skin and by morphologic studies of several tissues from the aborted fetus. Electron microscopic studies of the fetal tissues and cultured fibroblasts had large numbers of typical inclusions of I-cell disease, thus substantiating the diagnosis and intrauterine manifestation of the disease. The results indicate that prenatal diagnosis of I-cell disease is possible with enzyme assays of amniotic fluid and in cultures of fetal cells from the fluid. Enzyme studies of amniotic fluid can provide a preliminary diagnosis within a few hours, but it is suggested that the definitive diagnosis should be based on assays in cultured cells from amniotic fluid.


Growth of calcium oxalate in gel systems.

Methods are described for growing calcium oxalate in silica and gelatin gels under different conditions. The results obtained indicate that, in silica gel, calcium oxalate grows into single individual crystals, twins, and rosettes. Bipyramidal calcium oxalate dihydrate crystals similar to those present in the urine of stone formers were prepared in the silica gel system. The gelatin gel offered a suitably structured substrate on which calcium oxalate monohydrate crystals grow into aggregates. The orientation pattern of calcium oxalate crystals suggests that the growth process is controlled by the stereospecificity of the gelatin medium supporting growth.


Drug-biomolecule interactions: spin-probe study of effects of anesthetics on membrane lipids.

The electron spin resonance spectra of probes were used to study the organization and motion of molecules in hydrated stacked bilayer or liposome model membrane systems. The same steroid structures required for reduction of membrane permeability were required to produce well-ordered films of brain lipid. Alcohols and anesthetic agents influence the structure of model membranes, with their order of efficacy paralleling their pharmacological effectiveness. Spin probes were also used to demonstrate effects of calcium and local anesthetics on the rate of penetration of ascorbate into lipid bilayers.


Functional and structural changes in isolated rabbit liver mitochondria induced by fluorodichloromethane.

Fluorodichloromethane caused changes in substrate respiration rates and also in adenosine diphosphate (ADP)-stimulated rates of oxidation of both succinate and glutamate. Active swelling of mitochondria also occurred, but only in the presence of magnesium ion. ADP/O ratios were not significantly affected by the fluorocarbon. State IV respiration was stimulated, and the respiratory control ratio was linearly reduced with increased levels of fluorocarbon when succinate was used as substrate. Glutamate-stimulated respiration was greatly affected by the presence of the halocarbon but contrasted to the succinate responses. Low levels of fluorocarbon stimulated the state IV rate with glutamate whereas higher concentrations inhibited respiration. States III and IV respiration were completely inhibited by levels of 4.7 mg/ml of fluorodichloromethane when the nicotinamide adenine dinucleatide (NADH)-linked substrate was used. The NADH-linked substrate was more sensitive to the fluorocarbon treatment than the flavoprotein-linked substrate. The respiratory control ratio reached 1.0 at one-half the concentration required to elicit the same response with the flavoprotein-linked substrate, which suggested that NADH-dehydrogenase was inhibited. Active swelling of mitochondria occurred in the presence of magnesium and fluorocarbon, greatly exceeding controls, suggesting a loss of structural integrity and osmoregulation capacity. Increased rates of reduction of exogenously added cytochrome c also suggested changes in membrane integrity at higher fluorocarbon site of action of this compound is probably not the NADH-dehydrogenase system but the membrane system per se. Structural changes in the membrane system could account for the reported respiratory inhibition.


Effects of chlorimipramine and lysergic acid diethylamide on efflux of precursor-formed 3-H-serotonin: correlations with serotonergic impulse flow.

The effects of d-lysergic acid diethylamide (LSD) and chlorimipramine (CIMI) on the firing rate of serotonergic (5-HT) neurons and on the in vivo efflux of 5-HT were investigated in parallel. A cerebroventricular perfusing technique was used to measure the efflux of 3-H-5-HT formed in vivo from 3-H-tryptophan. Impulse flow in serotonergic neurons was monitored by single unit recording from raphe (5-HT) neurons. Doses of LSD and CIMI, which caused a similar degree of inhibition of raphe cell firing, were found to affect 5-HT efflux differently. LSD, at both the 75, and 150 mug/kg doses, produced a similar decrease in 3-H-5-HT efflux. In contrast, CIMI at a low dose (5 mg/kg) did not reduce 5-HT efflux, despite an inhibition in impulse flow. At a high dose (20 mg/kg), CIMI produced an increase in 3-H-5-HT efflux. We conclude that 1) LSD decreased 3-H-5-HT efflux by directly inhibiting impulse flow in 5-HT neurons and/or by a local effect on 5-HT terminals and 2) a low dose of CIMI produces no net change in 3-H-5-HT efflux because a reduction in impulse flow-dependent 5-HT release compensates for blockade by CIMI of 5-HT reuptake.


Correlation between structure, behavioral activity and rate of biotransformation of some ACTH4-9 analogs.

The effect of substitutions in adrenocorticotropin (ACTH4-9) on extinction of pole-jumping avoidance behavior in intact rats was investigated systemically at two-dose levels. Simultaneous introduction of 4-methionine sulfoxide and 5-D-lysine, in combination with 9-phenylalanine, led to a 1000-fold increase in behavioral potency. The same substitutions induced a 1000-fold decrease in melanocyte-stimulating hormone activity. Incubations of 14-C-labeled ACTH4-9 analogs, prepared by reductive methylation, were carried out with plasma and brain extracts. The resulting metabolites were separated by paper electrophoresis and paper chromatography. The concentrations of nonmetabolized hexapeptides, which appeared to be almost entirely responsible for behavioral activity, were determined as a function of incubation time. The in vitro half-life of intact hexapeptides correlated with their behavioral activity. Therefore, the increase in behavioral potency as a result of amino acid substitutions can be explained, at least partly, by increased resistance against biotransformation.


Cannabinoids: influence on neurotransmitter uptake in rat brain synaptosomes.

We have examined the effect of Delta1-tetrahydrocannabinol (delat1-THC) and 12 of its derivatives on the uptake of 3H-labeled norepinephrine (NE), dopamine (DA), serotonin (5-HT) gamma-aminobutyric acid (GABA) into synaptosomes in homogenates of various regions of rat brain. Delta1-THC inhibits the accumulation of NE and 5-HT into hypothalamic preparations and DA into the corpus striatum with Ki values of about 12 to 25 muM while GABA uptake into cerebral cortical preparations is inhibited less (Ki = 140 muM). The affinities of delta6-THC, 7-hydroxy-delta1-THC, 7-hydroxy-delta6-THC and cannabidiol for 5-HT, NE and GABA transports are similar to values for delta1-THC, while cannabigerol, cannabinol and delta6-THC-7-oic acid have substantially less affinity. Thus, hydroxylation of C-7 in delta6-THC does not alter inhibitory potency, but its oxidation to an acid and aromatization of ring A greatly reduce affinity. The hydroxyl at C-3(1) of ring C is critical for inhibition of NE, 5-HT and GABA uptake, since its acetylation or methylation abolishes activity. Inhibition of NE, DA, 5-HT and GABA uptake by all cannabinoids examined is noncompetitive. Only about 1% of delta1-THC and delta6-THC and 5% of cannabidiol are fully soluble under our experimental conditions.


Noradrenergic sensitivity of the cerebral cortex after chronic ethanol ingestion and withdrawal.

The effects of both chronic ethanol treatment and 3 days of ethanol withdrawal on the cyclic adenosine 3':5'-monophosphate (cAMP) response of cerebral cortical slices to norepinephrine (NE) were studied. ED50 cAMP responses for each group were determined using graded doses of NE. Chronic ethanol fed rats displayed subsensitivity to NE. There was a 4.3-fold shift of the dose-response curve to the right. Rats chronically fed ethanol and then put on 3 days of ethanol withdrawal developed supersensitivity. There was a 2.4-fold shift of the dose-response curve to the left. There was no difference in the maximal cAMP response observed in either the chronic alcohol or the chronic alcohol three day withdrawal experiments. It is possible that the biphasic modifications of effectors by ethanol ingestion and withdrawal from a subsensitive to a supersenitive is the result of one basic phenomenon.


Characterization of neuromuscular blocking action of piperidine derivatives.

cis and trans forms of 2-methyl-6-n-undecyl piperidines (C-11) are the main constituents of fire ant venom and have been studied for their mechanism of action on the neuromuscular transmission of the nerve-sartorius muscle preparation of frogs. At low concentrations (l times 10(-6)-2 times 10(-5) M), cis- and trans-C-11 irreversibly decreased the amplitude of spontaneous miniature end-plate potentials nerve-evoked end-plate potentials and iontophoretically induced acetylcholine depolarizations without changing the membrane potential. The quantal content and the focally recorded action potentials of nerve terminals remain unchanged. It was concluded that the piperidine derivatives block neuromuscular transmission postsynaptically through a decrease in the sensitivity of the end-plate membrane to acetylcholine. Pretreatment of the end-plate with d-tubocurarine did not effect the blocking action of trans-C-11. In direct binding experiments, trans-C-11 did not compete for the sites occupied by alpha-bungarotoxin, decamethonium, carbamylcholine and d-tubocurarine. These data suggest that trans-C-11 does not bind to the acetylcholine receptor site where the other cholinergic ligands have their affinity. It appears that these piperidine derivatives interfere with the coupling mechanism between acetylcholine-receptor binding and ionic conductance increases.


Changes in mechanical events and adenosine 3',5'-monophosphate levels induced by enantiomers of isoproterenol in isolated rat atria and uteri.

Beta adrenergic receptors of rat atria and uteri were examined with the use of enantiomers of isoproterenol as agonists and mechanical responses and adenosine 3',5'-monophosphate (cyclic AMP) levels as measured effects. Assuming that stereoselectivity reflects the unique asymmetry of receptors, potency differences between the enantiomers are expected to provide a sensitive indication of ligand binding. All effects in each tissue were investigated under similar experimental conditions. Both isomers produced the same maximum effect on all measured responses. Enantiomeric potency differences (in log units) for positive chronotropic and inotropic responses and increases in cyclic AMP levels in atria were 3.31, 3.51 and 3.48, respectively. In uteri, the values for reduction of spontaneous contractile amplitude and increases in cyclic AMP were 2.90 and 2.79 log units, respectively. Even though these absolute values varied slightly with the experimental conditions, they were consistently smaller in uteri than in atria. In both tissues, dose-response curves for production of mechanical effects were greater than 2 log units to the left of those for increases in cyclic AMP levels. Regardless of the interpretation of this phenomenon, the results show the following. 1) The stereoselectivity for isoproterenol-induced effects is different between the two tissues at both levels of response. Therefore, it is suggested that this reflects dissimilar beta adrenergic receptor types in rat atrium vs. rat uterus. 2) The stereochemical selectivity for isoproterenol-induced mechanical effects and increases in cyclic AMP is the same in rat atrium and in rat uterus. Therefore, the data support the postulate that cyclic AMP is formed from interaction of isoproterenol with a receptor that is similar to the one activated to produce a mechanical effect.


Heterosynaptic facilitation in the giant cell of Aplysia.

1. Heterosynaptic facilitation, defined as an increase of the efficacy of synaptic transmission between a test interneurone and a post-synaptic neurone, produced by the stimulation of a separate pathway, was studied in the left pleural ganglion. The experimental procedure consisted of detecting the effects of a brief tetanus, applied to tentacular and tegumentary nerves, on the amplitude of monosynaptic and unitary post-synaptic potentials (p.s.p.s) recorded in the left giant cell and generated by stimulating the test interneurone every 10 sec. The membrane potential of the test interneurone was simultaneously recorded. 2. Following heterosynaptic stimulation, the amplitude of the test p.s.p. increased, after a delay of about 30 sec, up to 250% of its original size; this increase subsided after 2-3 min or more. 3. Only the interneurones producing in the giant cell the e.i.p.s.p. (excitatory-inhibitory post-synaptic potential) were affected by hetero-synaptic facilitation. Other interneuronal types showed no changes in their synaptic transmission on the giant cell after heterosynaptic stimulation. 4. Heterosynaptic stimulation did not produce either orthodromic or antidromic spikes in the test interneurones clearly indicating that facilitation of test p.s.p. did not result from increased spike activity in the test interneurone. 5. Often heterosynaptic facilitation of the test p.s.p. was observed due to spontaneous activity in the heterosynaptic pathway, demonstrating the normal occurrence of the phenomenon. 6. Iontophoretic injection of 5-HT at critical, presumably synaptic, sites in the neuropil, evoked a facilitation of the test p.s.p. similar to heterosynaptic facilitation. Only the e.i.p.s.p.s. were so affected by 5-HT. On the contrary, other p.s.p. types were depressed by 5-HT as a result of conductance changes in the left giant cells. 7. Both heterosynaptic facilitation and 5-HT facilitation were suppressed by the presence in the bath of 5-HT (10(-5) M) and of LSD-25 (3 X 10(-4) M). The action of injected 5-HT on the membrane conductance of the left giant cell was also depressed in the pressence of 5-HT in the bath, but was unaffected by LSD-25 (3 X 10(-4) M). 8. From the parallelism of properties of heterosynaptic and 5-HT facilitation, it is suggested that 5-HT is the probable transmitter mediating heterosynaptic facilitation. It seems likely that 5HT is released from the heterosynaptic pathway at the level of the synaptic ending of the test interneurone on to the giant cell and that it increases the efficacy of this synapse, probably acting on the quantity of synaptic transmitter liberated.


The possible role of cyclic AMP in the neurotrophic control of skeletal muscle.

1. Motoneurones provide trophic control of some of the functional characteristics of skeletal muscle fibres. This study has been designed to test whether the adenylate cyclase: cyclic AMP system may offer one potential mechanism for the mediation of neurotrophic regulation. 2. The concentration of cyclic AMP was measured at various intervals after muscle denervation. Muscle cyclic AMP concentration increases for the first 2 days after nerve section. It reaches a maximum value at 48 h and subsequently returns to the control value at 7 days. 3. Cyclic AMP concentration is unchanged by muscle disuse for the first 3 days following limb immobilization. Four days after immobilization, however, cyclic AMP increases in both the disused and contralateral control muscles. This phenomenon has been tentatively ascribed to some aspect of the inflammatory response. 4. Changing the level of nerve section, and therefore the length of the residual nerve stump, changes the temporal pattern of the increase in muscle cyclic AMP concentration. 5. Reinnervation of a denervated muscle produces a decrease in muscle cyclic AMP concentration. 6. It is concluded from the results that some aspect of nerve function provides trophic regulation of the muscle adenylate cyclase: cyclic AMP system. The mechanisms by which this regulation may be applied are considered in the Discussion.


Pancreatic acinar cells: effect of acetylcholine, pancreozymin, gastrin and secretin on membrane potential and resistance in vivo and in vitro.

1. Intracellular recordings of membrane potential and input resistance have been made in vivo and in vitro from the exocrine acinar cells of rat pancreas using indwelling glass micro-electrodes. 2. The resting cell membrane potential and input resistance in the in vivo experiments were not markedly different from the values obtained in the in vitro experiments. The effect of both acetylcholine (ACh) and pancreozymin (CCK-Pz) on the pancreas in vivo as well as in vitro was to reduce both the acinar cell membrane potential and the input resistance narkedly. The amplitude of the evoked depolarization and the change in input resistance evoked by supramaximal stimuli were of the same magnitude in both types of preparations. 3. Gastrin had an effect on the acinar cell potential and resistance which was  indistinguishable from that of CCK-Pz or ACh. The effect of gastrin or CCK-Pz was, in contrast to that of ACh, not influenced by the presence of atropine. The reversal potential for the gastrin evoked potential change was about -20 mV. 4. Secretin in doses producing maximal volume secretion in vivo had no effect on acinar cell membrane potential and input resistance. 5. Dibutyryl cyclic AMP (5mM) and cyclic GMP (1mM) had no effect on cell membrane potential or resistance. 6. It is concluded that the in vitro superfused pancreas segment preparation is a useful model system in electrophysiological studies since it functions essentially as the in vivo preparation. In contrast to both gastrin and CCK-Pz, secretin has no effect on the bioelectrical properties of the acinar cells, indicating that there are no physiologically important secretin receptors in rat acinar cells.


The action of ether and methoxyflurane on synaptic transmission in isolated preparations of the mammalian cortex.

1. The actions of ether and methoxyflurane on the evoked potentials of in vitro preparations of the guinea-pig olfactory cortex were studied. Following stimulation of the lateral olfactory tract (l.o.t.) evoked potentials could be recorded from the cortical surface; these potentials consisted of an initial wave (the compound action potential of the l.o.t.) followed by a negative field potential which was associated with the synchronous excitation of many superficial excitatory synapses (population e.p.s.p.). Superimposed on the population e.p.s.p. was a number of positive peaks. These positive peaks reflect the synchronous discharge of many neurones and so have been called population spikes. 2. When ether or methoxyflurane was added to the gas stream that superfused the surface of the preparations, the population e.p.s.p.s. and population spikes were depressed at lower concentrations than those required to depress the compound action potential of the afferent fibres. 3. The evoked activity of individual cells in the cortex was depressed by ether and methoxyflurane. However, five of the twelve cells tested in ether showed an increase in their evoked activity at concentrations below 4-5%, but at higher concentrations these cells also became depressed. 4. Both ether and methoxyflurane depressed the sensitivity of cortical neurones to iontophoretically applied L-glutamate and may similarly depress the sensitivity of the post-synaptic membrane to the released transmitter substance. 5. Neither anaesthetic appeared to increase the threshold depolarization required for nerve impulse generation. Thus, the decrease of the discharge of the post-synaptic cells was primarily caused by a depression of chemical transmission. 6. Ether caused some cells in the cortex to alter their normal pattern of synaptically evoked discharge and both anaesthetics induced similar changes during excitation by glutamate.


The control of adaptive hypertrophy in the salt glands of geese and ducks.

1. Factors controlling adaptive hypertrophy, which occurs when marine, or potentially marine, birds drink salt water, have been investigated in geese and ducks using changes in salt-weight weight, RNA and DNA contents as indices of this process. 2. Unilateral post-ganglionic denervation in geese prevented the changes in [RNA] and [RNA]:[DNA] that occurred in the intact gland of birds given salt water for 24 hr; denervation had no significant effect in birds on fresh water throughout. 3. Atropine treatment also prevented the adaptive changes in geese given salt water. 4. In ducks give 0.3 M-NaCl for 48 hr salt-gland weight, [RNA] and [RNA]:[DNA] increase markedly. Treatment of ducks drinking fresh water with large doses of corticosterone and mammalian ACTH for 48 hr had no significant effects on salt-gland weight, RNA or DNA; mammalian prolactin treatment for 48 hr significantly raised [RNA]. 5. No changes in the total amount of DNA in the glands were observed in these experiments, thus indicating that hyperplasia does not occur within 48 hr of a bird first drinking salt water. 6. It is concluded that adaptive hypertrophy is controlled by secretory nerves, and that hormones, if they play any part in this process, have a permissive or secondary role. It is suggested that hypertrophy and the maintenance of the secretory cells in the fully-adapted state may be obligatorily related to secretory activity induced by cholinergic secretory nerves.


Electrophysiological properties of spinal motoneurones of normal and dystrophic mice.

1. The properties of spinal motoneurones of normal and dystrophic mice (129/ReJ) were examined with intracellular electrodes. 2. The following parameters of spinal motoneurones showed no significant differences between normal and dystrophic mice: resting and action potentials, the amplitude and duration of after-hyperpolarization, rheobasic current for excitation, threshold for excitation of the somadendritic membrane (IS-SD inflexion) and input resistance. 3. The changes in motoneurone properties observed 13-16 days after section of the sciatic nerve (axotomy) were similar in both normal and dystrophic mice. 4. The axonal conduction velocity of motoneurones in dystrophic mice was about ten times slower than that in normal mice. The conduction velocity of the sciatic nerve was only about 25% slower in dystrophic mice than in the normal animal. The estimated ventral root conduction velocity as well as the observed dorsal root conduction velocity in dystrophic mice was at least twenty times slower than that in normal mice. 5. In dystrophic mice, spinal motoneurones often showed multiple discharges in response to single, antidromic stimuli. The site of initiation of multiple discharge was located in the motor axon rather than in the motoneurone cell body. 6. In dystrophic mice, nerve impulses were transmitted from fibre to fibre ('cross-talk'). The site of impulse transmission among nerve fibres was near the distal portion of the spinal roots. 7. Synaptic potentials and peripheral reflex discharges evoked by stimulation of the dorsal roots showed a longer latency and were more prolonged in dystrophic mice than in the control mice. 8. The motoneurone properties of dystrophic mice showed no tendency of progressive changes with age ranging from 63 to 148 days. 9. It is concluded that the properties of motoneurone cell bodies examined in dystrophic mice are indistinguishable from those in normal mice and that the only abnormality in motoneurones of the former residues in the motor axon. 10. It is suggested that integrity of the discharge pattern of spinal motoneurones in dystrophic mice is interfered by anomalous impluse transmission in the motor axons and that the motoneurones in dystrophic mice are a homogeneous group rather than a mixture of "normal" and "abnormal" neurones.


Extracellular potassium changes in the spinal cord of the cat and their relation to slow potentials, active transport and impulse transmission.

1. By means of K-specific double-barrelled micro-electrodes the time course of changes in K+ concentration in the extracellular space of the lumbar spinal cord was examined after peripheral tetanic stimulation and after a single volley in a mixed peripheral nerve in non-anaesthetized, intercollicularly decerebrated and spinalized cats. 2. Tetanic stimulation (100 Hz) which increases the [K]e from 3 to 9 mM is followed by a phase of reduced [K]e during which [K]e decreases by 0.5 mM below resting level, lasting 1-2 minutes before returning to its original resting level. Evidence is presented that this subnormal phase of [K]e reflects active processes redistributing accumulated K+ from extracellular space. 3. The subnormal phase of [K]e can be registered only when the microelectrode is located in very close vicinity of discharging neurones and is not primarily dependent on the absolute level of increased [K]e. This can be considered as evidence that the neurones and not the glial cells are responsible for active reabsorption of K+ from the extracellular space. 4. Increased E1K]e is reflected in focally recorded potentials as a negativity and decreased [K]e as a positivity. The latency of focally recorded positivity is, however, shorter than the latency of reduced [K]e. This makes it likely that the positivity reflects not only passive hyperpolarization of glial elements, but also an active, electrogenic ion transport across neuronal membrane. 5. The shortest latency of increased [K]e induced by a single volley in a mixed peripheral nerve was found to be 9 msec; the peak, representing 0.5 mM, was attained after 40 msec and the total duration was 200 msec. A theoretical consideration is put forward that the time course of transient increase in [K]e is consistent with the suggestion that K+ which accumulates in the spinal cord after neuronal discharge is responsible for primary afferent depolarization. 6. Evidence is presented that increased [K]e, induced by a long lasting peripheral stimulation, is accompanied by decreased efficacy of impulse transmission.


Depression in the excitability of relay cells of lateral geniculate nucleus following saccadic eye movements in the cat.

1. The excitability of relay cells of the lateral geniculate nucleus during a saccadic eye movement was studied in alert cats. Excitability was assessed by the firing probability of the cells in response to electrical stimulation of the optic chiasm. Modifications in the excitability were evaluated during the period following eye movements, by triggering a stimulator from potential shifts in electro-oculogram and altering delays in the stimulus pulse. 2. The cells were classified into S and T cells, based on their response properties and the latencies to chiasmatic stimulation. With a saccade in a stationary patterned field, T cells showed a burst discharge, while the discharges of S cells were completely suppressed. 3. The excitability was depressed in both S and T cells for 150-200 msec after a saccade, when the eye movement occurred in light. However, the depression did not occur in complete darkness. 4. The depression occurred also in the absence of eye movement, when the patterned visual field was moved in a saccadic fashion. 5. The depression in S cells occurred during an inhibitory period. Since S cells do not receive signals on image movement directly from the retina, the depression was due to a recurrent inhibition by signals transferred through the T ganglion-relay cell channel. 6. The depression in T cells occurred concomitantly with the burst discharge. Since the recurrent inhibition was operating less effectively during the period, the depression may be due to a phasic occlusion of the test impulse by coincident high-rate firings in the same cell.  7. The impairment in transmission of visual information through the lateral geniculate nucleus during the period following eye movements has been discussed in connexion with a neurophysiological basis for saccadic suppression.


Five seasons of sadness.

In response to her grandmother's death, moving back to her family, and beginning kindergarten, Bridget experienced a very difficult adjustment. During the period of play therapy, she began grief work: "Emancipation from bondage to the deceased, readjustment ot the environment in which the deceased is missing, and formation of new relationships."15 Bridget, at the age of five, has experienced five seasons of sadness. Yet her quiet and determined strength has enabled her to readjust to her once threatening environment. The therapist suggested continued therapy in the school environment to aid Bridget in formation of new relationships. The shadow of Bridget's five seasons of sadness seems to have lifted. Her bright affect and look of expectancy seem to be asking of the other children: "Will you be my friend? For no reason that I know Except I want you so."16


Assuring accountability for the poor: patient advocates.

The three patient advocate groups that existed in this particular state hospital and differing structures, functions and degrees of effectiveness. However, all three seemed necessary to assure that the serving institution and its professionals would become accountable for the success or failure of their methods and performance. Two of the groups (relatives group and legal aid group) seemed to be functioning effectively within certain limits which each thought to be appropriate in advocating patient causes. Thus, they were providing the countervailing force that is a necessary component of accountability when the persons to be served are a powerless group. The consumer group being the powerless group was not effective in advocating its own cause. It would have been very beneficial to this group had they succeeded. There is a therapeutic effect of having control over one's own life that fosters independence and decreases withdrawal and feelings of alienation from the social order. Certainly these are the goals of the mental health professionals for their patients. In order to realize these goals certain changes would have to be made in the mental health code would have to be changed to include formalized grievance procedures and the desirability of consumer groups would have to be formally established. If this were so, consumers of mental health services might be more effective in advocating their own causes and would be more able to hold the serving institution accountable


[A case of carcinoma of the stomach with calcification (author's transl)].

The authors found only 25 cases of calcified carcinoma of the stomach in the literature. The course in this case was rapidly fatal. Calcification within the tumour infiltrates the gastric wall and has a very characteristic miliary appearance, permitting one to make the diagnosis on a straight X-Ray of the abdomen. The mechanism of precipitation of calcium within certain mucus-secreting carcinomas has not been determined. Various theories are suggested.


Clinical diagnosis of breast cancer.

Breast cancer is the most common malignant neoplasm in women, and 6% will develop it during their normal life expectancy.  There is a group who have a high risk of developing breast cancer.  The recent improvement in cure rates seems to be jue chiefly to earlier diagnosis rather than to improved methods of therapy.  The physician, by careful periodic breast examinations and by the judicious use of diagnostic aids such as mammography and thermography, especially in the high risk group, has a golden opportunity to pick up cancer in a localized stage where the prognosis for cure with appropriate therapy is excellent.  A tentative diagnosis of breast cancer (Table XI) can be made with a fair degree of accuracy by taking a careful history, utilizing and combining available statistics about the frequency, median age, characteristic symptom complexes of the common breast lesions and factors related to a high mammary carcinoma risk, and by a systematic and thorough breast examination supplemented with diagnostic aids when appropriate.  However, biopsy and histologic examination is mandatory in all patients with a) true, three dimentional, dominant lumps even if diagnostic aids are negative except for cysts which can be safely aspirated under controlled conditions; b) suspicious lesions found by diagnostic aids even though there are no clinical findings; c) serous, serosanguineous, bloody, or watery nipple discharge; and d) other signs of cancer, i.e. eczema of the nipple, axillary adenopathy, etc., in order to determine with absolute accuracy whether the lesion is benign or malignant.


A systems analysis approach to planned change in a clinical psychiatric program.

In health car facilities, systems studies are commonly conducted as ivory-tower operations with minimal impact and little practical result. A real-life case study was presented to demonstrate how it can be used effectively in a collaborative manner to implement constructive change. Systems Analysis techniques were succe-sfully used with the staff of a psychiatric ward to plan for a satellite outpatient treatment program. The focus was on collaboration to help the ward to obtain objective information for decisions concerning the need for changes in their existing system.


Adenosine cyclic 3',5',-monophosphate phosphodiesterasr inhibitors. 2.3-Substituted 5,7-dialkylpyrazolo [1,5-a]pyrimidines.

A number of 3-bromo-, 3-nitro-, and 3-ethoxycarbonyl-5,7-dialkylpyrazolo[1,5-a]pyrimidines were synthesized and screened as in vitro cAMP phosphodiesterase inhibitors. The condensation of 3-aminopyrazole with symmetrical beta-diketones (acetylacetone, heptane-3,5-dione, etc.) afforded symmetrical dialkylpyrazolo[1,5-a]pyrimidines (5). The reaction of 3-aminopyrazole with unsymmetrical beta-diketones (hexane-2,4-dione, heptane-3,5-dione, etc.) gave a mixture of 5-methyl-7-alkylpyrazolo[1,5-a]pyrimidine (3) and 5-alkyl-7-methylpyrazolo[1,5-a]pyrimidines (4). The technique for the separation of 3 from 4 is described. The inhibition constants, alpha (the ratio of the molar I50 of theophylline to the molar I50 of the test compounds), were subjected to a Hansch correlation analysis. The results indicated that PDE isolated from beef heart tissue was most sensitive to changes in the length of the alkyl group in the 5 position of the pyrazolo[1,5-a]pyrimidine ring, whereas the PDE isolated from rabbit lung tissue was more sensitive to changes in the length of the 7-alkyl group. Experimentally and theoretically, the n-propyl group was found to approximate the ideal size for the alkyl group in both the 5 and 7 positions;5,7-di-n-propyl-3-ethoxycarbonylpyrazolo[1,5-a]pyrimidine (5e) was the most potent inhibitor of both lung and heart PDE.


Synthesis and antitumor activity of 5'-phosphates and cyclic 3',5'-phosphates derived from biologically active nucleosides.

Syntheses and biological activities of 12 N6-substituted adenosine 5'-phosphates and 15 cyclic 3',5'-phosphates are described. Included among these are the cyclic phosphates of the naturally occurring anticodon adjacent modified nucleosides, N6-(delta2-isopentenyl)adenosine and N-(purin-6-ylcarbamoyl)-L-threonine ribonucleoside. Also reported in this paper are the 5'-phosphates and cyclic phosphates of the cytokinins, N6-benzyladenosine, kinetin ribonucleoside, 3-(chloro-trans-2-buten-2-yl)adenosine,6-o-chlorophenylureidopurine ribonucleoside, and 6-allylureidopurine ribonucleoside. The 5'-nucleotides were prepared by direct phosphorylation of the corresponding ribonucleosides with POCl3 and triethyl phosphate. These compounds were converted to the cyclic 3',5'-phosphates by cyclization of the corresponding 5'-nucleotides with dicyclohexylcarbodiimide. Comparison of the cytotoxicity of the ribonucleosides with their 5'-nucleotides and cyclic 3',5'-nucleotides showed that some of the 5'-phosphates and cyclic phosphates were almost as active as the parent nucleosides. The 5'-nucleotides and the cyclic phosphates were more soluble than the parent nucleosides. The cyclic 3',5'-nucleotides were examined as alternate activators of cAMP-dependent protein kinase from beef heart. While all of the analogs studied showed some activity toward this enzyme, several compounds were more effective than cAMP itself. The analogs were also tested as substrates for cyclic 3',5'-nucleotide phosphodiesterase from beef heart. The N6-alkyl-cAMP analogs were poor substrates for the enzyme, while N6-carbamoyl-cAMP derivatives were inert toward this enzyme. These compounds did not inhibit the phosphodiesterase. Some of the cyclic phosphates exhibited marginal effect in the inhibition of glycogen synthesis in skin slices.


Synthesis and biological activity of 4-beta-Iribofuranosyl-1,3-dihydroxybenzene ("1,3-dideazauridine").

In view of the marked antitumor activity of 3-deazauridine, the synthesis of 4-(beta-D-ribofuranosyl)-1,3-dihydroxybenzene (1,3-dideazauridine) and its dibenzyl derivative was carried out. 4-Bromo-1,3-dihydroxybenzene was converted to its dibenzyl derivative, which, upon reaction with n-butyllithium followed by treatment with anhydrous cadmium chloride, gave bis(1,3-dibenzyloxyphenyl-4)cadmium. Condensation of this intermediate with 2,3,5-tri-O-benzoyl-D-ribofuranosyl chloride in refluxing toluene, and subsequent removal of the protecting benzoyl groups, afforded 4-(beta-D-ribofuranosyl)-1,3-dibenzyloxybenzene which, upon catalytic hydrogenation over Pd/C, furnished the desired 4-(beta-D-ribofuranosyl)-1,3-dihydroxybenzene. The beta configuration at the anomeric center was established by NMR and hydrogen bonding studies. 4-(Beta-D-ribofuranosyl)-1,3-dibenzyloxybenzene inhibited the growth of leukemia L1210 cells by 50% at 7 x 10(-6) M, and that of mammary carcinoma TA3 cells at 5 x 10(-5) M. Dideazauridine itself was less active, inhibiting the leukemia L1210 but not the TA3 cells at 1 x 10(-4) M, but the compound was significantly active against herpes simplex (type I) virus in vitro.


Probiotics. Antistaphylococcal and antifibrinolytic activities of omega-amino-and omega-guanidinoalkanisulfonic acids.

A series of omega-aminoalkanesulfonic acids (1-5) and omega-guanidinoalkanesulfonic acids (6-9) has been tested for antistaphylococcal and antifibrinolytic activities. Most of the omega-aminoalkanesulfonic acids and 6 produced better protection against Staphylococcus aureus infections in mice than gamma-aminobutyryl-L-histidine. Compound 4 was the best antistaphylococcal agent among the omega-aminoalkanesulfonic acids and compound 6 among the omega-guanidinoalkanesulfonic acids. Most of the omega-aminoalkanesulfonic acids have antifibrinolytic activity, while none of the omega-guanidinoalkanesulfonic acids has significant antifibrinolytic activity. Compound 4 possessed the highest antifibrinolytic activity which was equal to or greater than that of epsilon-aminohexanoic acid.


Synthesis and biological activity of some 8-substituted selenoguanosine cyclic 3',5'-phosphates and related compounds.

8-Bromoguanosine cyclic 3',5'-monophosphate, 8-bromoguanosine 5'-monophosphate, and 8-bromoguanosine served as intermediates for the chemical synthesis of a series of 8-substituted seleno cyclic nucleotides, nucleotides, and their nucleosides. Selenourea was found to be a useful reagent in synthesizing these seleno-substituted nucleoside, nucleotide, and cyclic nucleotide. A nucleic acid analyzer was used to study the hydrolysis of these cyclic nucleotides by phosphodiesterase. It was found that all of the 8-substituted selenoguanosine cyclic 3',5'-phosphates synthesized, except 8-MeSe-cGMP, were resistant to hydrolyze by phosphodiesterase. These 8-substituted seleno cyclic GMP derivatives showed some antitumor activities against murine leukemic cells (L5178Y) in vitro and in vivo.


Synthesis and antitumor properties of new glycosides of daunomycinone and adriamycinone.

The synthesis of 4'-epi-daunorubicin and of 4'-epi-adriamycin was performed by condensation of 2,3,6-trideoxy-3-trifluoroacetamido-4-O-trifluoroacetyl-alpha-L-arabino-hexopyranosyl chloride with daunomycinone or the protected adriamycinone derivative 17, respectively. Both the alpha and beta anomers were obtained and characterized. All new compounds are biologically active in cultured cells and the alpha anomers display noticeable activity in experimental tumors in mice. Interestingly, 4'-epi-adriamycin (4) appears nontoxic to cultured heart cells up to a concentration of 5 mug/ml.


Effect of 2-deoxy-D-glucose on herpesvirus-induced inhibition of cellular DNA synthesis.

In pseudorabies virus-infected cells host DNA synthesis is turned off 4 to 5 h postinfection. In the presence of 0.5 mM 2-deoxy-D-glucose, however, synthesis of both cellular and viral DNA proceeds unimpaired throughout the virus replication cycle. The uptake of radioactive thymidine into mock-infected cells is not altered in the presence of 2-deoxy-D-glucose. Virus-specific protein synthesis and particle formation also proceed in medium containing the deoxy sugar, but the virus particles produced are noninfectious and cell fusion is inhibited.