Microelectrophoretic application of antagonists of putative neurotransmitters onto various types of bulbar respiratory neurons. Seven antagonists of putative neurotransmitters were applied to bulbar respiratory neurons and, for comparison, also to unspecific cells. The antagonists exerted distinct effects when released alone, permitting to draw conclusions about receptor properties of the various cell types. With strychnine, specific antagonist of glycine, excitation prevailed in EI, I and E neurons. With bicuculline, specific antagonist of GABA, excitation preponderated in EI and E cells. About half of the unspecific neurons were activated and the remainder were unresponsive. GDEE (glutamatediethylester), antagonist of glutamate, excited part of the IE neurons and inhibited part of the E units, while the remainder of both types as well as 2 EI cells tested were not affected. With flupentixol, antagonist of dopamine, excitation prevailed in I neurons. About half of the IE and E units remained unaffected, while in the remainder E cells inhibition preponderated over excitation. With yohimbine, an alpha-adrenoceptor blocker, inhibition prevailed in E units. The two EI as well as the majority of the I neurons remained unaffected, with two cells of the latter type being activated. Propranolol, a beta-adrenoceptor blocker, inhibited about half of the E neurons, while the remainder as well as most IE and the 2 EI cells tested were not affected. Cyproheptadine, an antagonist of 5-HT, excited most E neurons. As concerns NE-receptors, those of the alpha-type might be involved in activation of part of the E cells only, whereas all other NE effects (inhibition or activation) are mediated by CNS-specific receptors different from the alpha- and beta-type. 5-HT effects apparently are mediated by two different receptor types.


Relation of colonial morphologies of strain of Streptococcus pneumoniae in soft-agar to the encapsulation. Among 40 fresh isolates of Streptococcus pneumoniae 12, 25 and 3 strains, respectively, exhibited large round, small round and compact colonial morphologies in soft-agar medium. Every large round strain possessed a capsule, almost half of the small round strains had capsules, while all of the large round type growth showed very high mouse virulence and 1.0 mg of these organisms was capable of absorbing a minimal amount of passive protective antibody in rabbit antiserum, prepared with the homologous strain, against challenge infection with homologous organisms in mice. Its variant showing compact type growth in soft-agar was mouse avirulent and a similar amount of the mouse passive protective antibody could not be absorbed with 100 mg of these organisms. These experimental results indicate that the soft-agar technique can be used for the identification of encapsulated strains of Streptococcus pneumoniae.


[Researches on some properties of inhibitory substances produced by strains of "N. meningitidis" (author's transl)]. Some properties of three inhibitory substances produced by as many strains of N. meningitidis have been studied. All these substances appeared to be completely destroyed by proteolytic enzymes (trypsin and pronase) and to be very heat-stable; they were insensitive to ultraviolet light and they were stable pH 2-10; filtration through filters of different porosity (Seitz, Berkefeld, millipore membranes) resulted in loss of the inhibitory effect constantly. On the basis of these results and on their specific action specific action, it is possible to class these substances among bacteriocins ("meningocins") at low molecular weight.


[Analysis of deoxyribonuclease I and II activity as a function of ionic strength and pH]. The action of deoxyribonucleases I and II has been studied as a function of ionic strength and pH, in the light of the theory of the ionic control of biochemical reactions (P. Douzou and P. Maurel (1977) Proc. Nat. Acad. Sci. USA, 74, 1013-1015). The pattern of DNA degradation by the two enzymes fits the general principles of the theory. However, the activity of DNAase II, a dimeric, basic protein (pI = 10,2) appears to be scarcely modulated by variables such as ionic strength and pH. This is reminiscent of what was elsewhere observed with the system double stranded RNA-seminal RNAase (also a very basic, dimeric enzyme), and could, therefore, tentatively be correlated with the dimeric and/or the very basic nature of the enzyme protein.


[Analysis of phenothiazine neuroleptics by means of non-aqueous titrimetry]. Phenothiazine neuroleptic drugs, either as pure compounds or in pharmaceutical dosage forms have been quantitatively assayed by nonaqueous titrimetry. Suggested procedure is applicable to basic drugs and their corresponding salts without prior cleanup overcoming the interferences due to excipients.


[Sympathetic mediation of induced and spontaneous activity of the pelvis-ureter in vitro]. The spontaneous activity of guinea-pig renal pelvis-ureter is regulated by the adrenergic system. Spontaneous rhythmic contractions, and contractions induced by Noradrenaline are inhibited by Dihidroergotamine and Phentolamine. Alpha-adrenergic blocking agents block also contractions induced by histamine, angiotensin and barium chloride, but not contractions induced by electric stimulation. The Authors suggest an hypothetical model for the activation of the adrenergic receptor: Noradrenaline (NE) recognition sites are activated only by NE, whereas complementary sites can be activated by NE or other agonists. Both sites are blocked by alpha-adrenergic blocking agents.


Persistent asthma after treatment with beta-blocking agents. Two atopic subjects with previously trivial asthma developed severe wheezing after receiving beta-adrenergic blocking agents. These drugs were discontinued but severe airways obstruction was still present more than 18 months later.


Antagonistic properties of N-methyl-N'-[2-(5-ethyl-4-imidazolyl-methylthio)ethyl]thiourea. In the present paper the Authors describe the synthesis and the biological effects of N-methyl-N'-[2-(5-ethyl-4-imidazolylmethylthio)ethyl]thiourea (etiamide). This compound appeared to inhibit, to a certain extent, the effect of histamine on H2-receptors. This inhibition, however, was less pronounced than that elicited by metiamide and in some of the preparations examined appeared to be of the non-competitive type.


[Postvaccinal encephalitis in adult. A case with anatomo-clinical report (author's transl)]. While post-vaccinal encephalitis in children is unfortunately well known, its occurrence in adults is rather exceptional. The calendar of a poly vaccination situated so close to each other, most probably account for such an evolution. The case reported concerns a man of 27 years who presented 10 days after a polyvaccination (7 vaccines in 2 months) altered consciousness, epileptic fits, right hemiplegia, vegetative dysfunctions and meningeal status. Evolutionary clinical course led to death in 21 days. Neuropathological findings showed lymphocytic infiltrates in the cortex and basal ganglia, congestive aspect with petechial areas in the white matter and inflammatory meningeal infiltrates. All viral investigations both in the blood, brain and cerebrospinal fluid remained negative. Post-vaccinal encephalitis is reviewed. Vaccination cannot be condemned, but in case of polyvaccination lapse of time in between vaccines should be longer so as to prevent such exceptional evolution and fatal course.


Filarial elephantiasis in French Polynesia: a study concerning the beliefs of 127 patients about the origin of their disease. 127 patients from Tahiti who were suffering from elephantiasis were interviewed about their opinion of the origin of their disease. Ancestral beliefs are still widely held even after 25 years of antifilarial campaigns which have resulted in a drastic decrease in endemicity with almost no clinical incidence. It is disappointing that the responsibility of mosquitoes is denied by a majority of patients. The explanations are to be found in the unusual evolution of this disease and in the small importance attached to sanitary education.


Evidence for a strong non-adrenergic component in the motor transmission to the rabbit vas deferens. Isolated vas deferens preparations from 16 rabbits of the New Zealand white strain were subjected to electrical and chemical excitability under physiological conditions and under the influence of drugs. Such smooth muscle fibres were strictly confined to the terminal 3 cm. segments of the distal 'urethral' portions of the vasa deferentia. Intermittent field stimulation, at 60 second intervals, was provided by a stimulator of low output impedance under constant parameters of voltage, pulse width and frequency. Results from this investigation revealed the undermentioned anomalous but distinct findings viz: (a) the presence of a minor adrenergic component which disappeared in phentolamine but remained unaffected by prolonged exposures to phenoxybenzamine; (b) the presence of a predominantly non-adrenergic component which was totally refractory to phenoxybenzamine but suffered a weak diminution in phentolamine; and (c) the picture of a phentolamine-insensitive but twitch-inhibiting effect shared by both tyramine and noradrenaline. Rabbit vasa consistently displayed a remarkable insensitivity to the motor effects of submicromolar concentrations of the putative neurotransmitter substance i.e. noradrenaline. The indirect sympathomimetic agent, tyramine failed to elicit any contractions when employed in doses as massive as 290 micrometers. i.e. 5 x 10(-5) g/ml, except in 2 out of 14 trials when a weak phentolamine-susceptible bursts of single contractions were produced. Thus, the adrenoceptors involved in the mediation of the twitch-inhibiting effects, appear not to behave towards conventional adrenoceptor antagonists in the classified manner. It seems appropriate therefore to invoke an unknown neurotransmitter for the non-adrenergic component in this motor transmission.


Acetylcholinesterase in the central nervous system and digestive gland of Achatina fulica Bowdich. Cholinesterase (ChE) activity was measured in the central nervous system (CNS) and in the digestive gland of the pestiferous land snail Achatinafulica Bowdich, by the method of Huegra et al. (1952). Acetylcholinesterase (AChE), and benzoylcholinesterase (BeChE) activity was higher in the former than in the latter. The complete inhibition of the enzyme activity with 10(-2) M eserine indicates that the ChE examined is AChE. The Km values of the AChE from the digestive gland and the CNS were 3.1 x 10(-5) and 9.0 x 10(-5) (M), respectively. The enzyme is the most active at pH 8.2 and 37 degrees C up to 60 min.


Tissue pH in the assessment of ischaemic muscle damage. Measurement in vivo of the pH of superficial muscles of dogs was found suitable for the assessment of the severity of ischaemic muscle damage and for the detection of irreversible lesions.


Spectral transitions in purple membranes from Halobacterium halobium. I. Effect of preliminary illumination on photochemical processes. The effect of preliminary illumination of purple membranes by yellow light on the difference spectra of short-lived intermediates has been studied. It has been found that changes of the optical density of two of these intermediates, which have the maxima of the difference spectrum at 412 nm and 650 nm, coincides well with the kinetics off the known reversible transitions of the main band of the purple membrane absorption (560 570), i.e., 13-cis-trans transitions. The changes at 412 nm and 650 nm are proportional to the concentration changes of the all-trans retinal and 13-cis retinal, respectively. It was concluded that the formation of the short-lived 412 and 650 intermediates occurs in different photochemical cycles. The pH is found to affect the formation of the 650 intermediate.


Oxidant-induced alterations in pulmonary microsomal mixed-function oxidation: acute effects of paraquat and ozone. Pulmonary mixed function oxidations are altered by in vivo exposure to paraquat or ozone. The cytochrome P-450-mediated benzphetamine N-demethylase activity was inhibited by low doses of both oxidants (paraquat, 20 mg/kg, ip; ozone, 1 ppm, 24 hrs). The effect of paraquat did not appear until 4 days after treatment and was still apparent after 2 weeks. The inhibition following exposure to ozone appeared immediately (1 day) and had resolved by one week. Cytochrome b5-mediated lipid desaturation was unaffected by the paraquat treatment and was stimulated by ozone. These alterations in pulmonary mixed function oxidase activities could not be explained by microsomal lipid peroxidation. The concentrations of cytochromes P450 and b5 also did not accurately reflect the altered enzymatic activities. The sites of the paraquat- and ozone-mediated alterations are still unknown.


Ion movements during energy-linked mitochondrial structural changes. The structure of isolated rat liver mitochondria has been observed in the electron microscope following incubation of the mitochondria in vitro under a variety of conditions. The results show that ultrastructural changes are only associated with the energization and deenergization of isolated mitochondria if the composition of the incubation medium permits ion movements in or out of the matrix. The mechanism of energy coupling does not appear to depend on these major mitochondrial structural changes. The addition of low levels of valinomycin greatly increases the rate at which the matrix compartment swells and shrinks on energization and deenergization even at low K+ concentrations.


[Attempts to characterize dexamethasone receptors present in rat kidney cytosol]. Density gradient centrifugation analysis has been used to study rat kidney receptors for dexamethasone. Density gradients performed with cytosol incubated with 3H-dexamethasone in a Tris 10 mM, EDTA 1.5 mM, glycerol 5%, monothioglycerol 12 mM (pH 7.5) buffer never gave any good evidence of a 85 radioactivity peak probably due to instability of the complex. The stability of the complex for at least 30 h was achieved by homogenizing the tissue and incubating cytosol in a distilled water, glycerol 5% monothioglycerol 12 mM solution. Density gradient centrifugation analysis in this way showed radioactivity peak at the bottom of the tube probably due to the aggregation of the complex. Since this drawback is inherent in the technique it must be concluded that density gradient centrifugation analysis cannot be used to study the dexamethasone-receptor complex of the kidney.


[Determination of chemoantibiotic levels in tissues and biological fluids (author's transl)]. On the premise that a microorganism is sensitive or resistant to a chemoantibiotic according to it's concentrations in biological fluids and tissues, the A. examines those methods available in chemoantibiotic titration such as chemical, enzymatic, radio-immunologic, turbidimetric, potentiometric and agar-diffusion methods. By further evaluating the limits and capacities of these different methods, the A. describes in larger detail the agar-diffusion method which is considered more sensitive, practical and efficient. A brief report is made on the possibility of measuring antibiotic association.


[Culture media for antibiotic sensitivity testing (author's transl)]. The medium is undoubtedly one of the most important factors influencing the sensitivity testing techniques. The possible role of pH and different components of media, as such as the concentration of peptone, tymidine or tymine, riboflavin, cations (MG++, Ca++, Fe++), mineral salts, agar and phosphate (added eventually with buffer solutions) is discussed. The conclusions point to the need of synthetic media which are suitable for growing all common pathogens and to not interfere with the antibacterial activity of drugs.


[Neurohumoral transmitter mechanisms at the cellular level]. The term neurohumoral transmission designates the transfer of a nerve impulse from a presynaptic to a postsynaptic neuron by means of a humoral agent e.g. a biogenic amine, an amino acid or a peptide. This process involves several steps, i.e. biosynthesis, storage, release, receptor interaction and inactivation of the transmitter. A neuromodulator modifies, for instance the release of a transmitter by action on a presynaptic transmitter neuron. Biogenic amines may also be released from non-synaptic nerve terminals and possibly exert a modulatory effect on other neurons. The regulation of the transmitter dynamics at the enzymatic level occurs by end-product inhibition and by enzyme induction. In addition, there are regulatory mechanisms originating from autoreceptors and postsynaptic receptors which operate via feedback action. Stimulation or inhibition of receptors may lead to receptor sub- and supersensitivity respectively. Numerous neuro-psychotropic drugs used in medical practice act on the various steps of neurohumoral transmission and thereby influence the dynamics of neurotransmitters and modulators.


[Central organization of transmitter systems]. The organisation of the central nervous system (CNS) is considered from the point of view of the chemical specificity of neurons with respect to their neurotransmitters. The mechanisms of neurotransmission are summarized as well as the approaches used to determine the neurotransmitters of specific CNS pathways. As an example, the neurotransmitters of the basal ganglion and their interactions are presented and their implications for the pathophysiology of Huntington disease and possibly schizophrenia are discussed.


[The morphologic diversity and complexity of synapses and microcircuits]. After mentioning the dendrite arborizations and the dendritic spines which represent an important site of synaptic interactions and neuronal plasticity, the main synaptic types, chemical and electrical, excitatory and inhibitory are mentioned and their morphological characteristics and dynamic properties are discussed. In the second part, the synaptic connections between several nerve cells, and the module principle of cerebellar and cerebral cortex are described. Significant progress is expected from the application of the 14C-deoxyglucose technique which is used as a histochemical marker of neuronal activity.


[Physiopathology of endogenous psychoses]. Metabolic theories of manic-depressive psychosis and schizophrenia are reviewed and constructivist models are presented which attempt to integrate biochemical, neurophysiological and clinical findings.


[Potential therapeutic activity of GABA-mimetic drugs in neuropsychiatry]. Several pieces of evidence support the view that GABA neurons inhibit the DA system both in extrapyramidal and limbic regions. This inhibition is exerted on cell bodies and terminals of DA neurons and is involved in the regulation of their activity. A recently synthesized GABAmimetic compound SL 76.002 has considerably helped in elucidating the role of GABA in this regulation as well as the therapeutic implication of changes of GABAergic transmission in human brain. Thus, impairment of dopaminergic transmission by SL 76.002 has been shown to be effective in iatrogenic extrapyramidal syndromes such as L-DOPA-induced involuntary movements in parkinsonian patients and neuroleptic-induced tardive dyskinesias: the first attributed to an exaggerated formation and liberation of DA, the second to supersensitivity of target cells of DA neurons. Furthermore, GABAergic medication has been confirmed to be useful in Huntington's chorea in which some symptoms originate from degeneration of striatal GABAergic neurons. Finally, GABAergic inhibition on cellular excitability has been proved to ameliorate epilepsy. However, SL 76.002, contrary to the expectation, was not effective in schizophrenia suggesting that GABA does not play a major role in the pathogenesis of this disorder.


[The development of psychopharmacotherapy]. Based on experiences during the longterm treatment with psychotropic drugs the following subjects are discussed: extrapyramidal side effects of neuroleptic drugs, cardiovascular side effects of antidepressants, side effects of Lithium, interactions with other medications, influence on laboratory tests and also the psychopharmacological treatment during pregnancy.


Acetyl-CoA carboxylase activities in blood and liver of chicks and their dependency on biotin status. Acetyl-CoA carboxylase activities from whole blood and, in some of the experiments, from liver were found to be lower in biotin-deficient chicks compared with controls. In vitro stimulation of liver acetyl-CoA carboxylase activity by biotin appeared to be a better index for the evaluation of the biotin status than measuring the enzyme activity alone.


Relative number and proliferation kinetics of hemopoietic stem cells in the mouse. A model calculation of the hemopoiesis of the mouse based on known hematologic data leads to the conclusion that approximately 3% of all nucleated bone marrow cells are stem cells (pluripotent plus committed stem cells). By a new 125IUdR labeling technique on radiation chimeras, a relative number of 2%-7% stem cells was determined. In previous studies with test systems for stem cells using colony formation in vivo or in vitro, a relative number of stem cells of at least one order of magnitude lower has been estimated. In this study the stem cells are found to have a turnover time of about 4.3 days in the donor mice. This turnover time remained unchanged even after transfusion of marrow cells into lethally irradiated recipient mice. Radiosensitivity determinations yielded a D0 of 80 rad for stem cells in S-phase and D0 of 185 rad for stem cells distributed throughout the entire cell cycle. The respective extrapolation numbers were 1.23 and 1.14. Experiments using an 3H-TdR suicide technique revealed different cell cycle parameters for bone marrow stem cells seeding to the spleens and to the femurs of lethally irradiated recipients, primarily a shortening of S-phase in cells seeding to femurs. The method described here provides a new approach to hematologic stem cell research.


Collection, storage and transfusion of blood stem cells for the treatment of hemopoietic failure. Migration of hemopoietic stem cells via the blood to sites of stem cell need is a principle that becomes established during the embryonic development of hemopoiesis and can be observed in the adult whenever bone marrow transplantations are being performed. The regular presence of stem cells in the peripheral blood lends itself to the study of their collection, storage, and use for transfusion purposes in cases of bone marrow failure. Both in dog and in man, granulocyte-macrophage progenitor cells (CFU-C) can be collected by leukapheresis from the blood in large quantities, particularly if the yield is increased by the administration of mobilizing agents such as dextran sulfate, and appear to be an indicator for the presence of stem cells. For collection and storage, a closed plastic bag system has been developed that allows the safe handling of the cells. The loss of CFU-C from freezing and thawing with DMSO as a cryoprotective agent is only 10%-20%. If frozen and thawed mononuclear leukocytes are transfused into 1200 rad whole-body X-irradiated autologous or allogeneic recipient dogs, a hemopoietic take is observed when 0.2 X 10(5) CFU-C are present among the mononuclear leukocytes (MNC). Graft-versus-host disease can be avoided in the allogeneic situation when a purified CFU-C rich cell fraction is being transfused. In man collection and storage of MNC including CFU-C is feasible and may eventually become a therapeutic tool.


Regional variability in circadian rhythmicity of intestinal digestive-absorptive functions. The circadian rhythms of sucrase, maltase, isomaltase, trehalase, lactase, gamma-glutamyltransferase, leucylnaphthylamide hydrolyzing activity, alkaline phosphatase and monosaccharide transport were assessed in each fifth of the small intestine of the rat in order to determine if an entire enzyme or transport system population responded in a similar manner or if there were regional differences. Animals were maintained under a light-dark cycle and fed from 1400-1800, EST for 7 days. Functional activities were assessed every 4 h for 24 h, inclusively. Quantitative, and in a few instances, qualitative differences in different areas of the intestine were found for all functions. There were portions of the lactase and alkaline phosphatase populations which displayed no rhythmicity in activity. When rhythmicity was observed there were differences in the activity patterns along the intestine for all functions. Thus, the rhythm patterns obtained from homogenates of the entire small intestine are a composite of the patterns in regions of high average activity. Also, there appears to be a reasonable amount of local control of the various functions.


[A study of gamma-glutamyl-transpeptidase and of the ratio of IgA to transferrin in the pregnant woman (and when using oral contraception) and in the newborn (author's transl)]. The authors confirm the absence of a marked variation in levels of gamma-glutamyl-transpeptidase in women under the influence of oestrogens, but a considerable rise in this enzyme in the newborn, particularly after delivery following vaginal obstetrical manoeuvre. They base these findings on 242 samples taken from pregnant women in the 3 trimesters of normal pregnancies and after delivery, and in the cord blood of newborn infants as well as in patients taking oral contraception. There is a slight rise in the levels of immunoglobulins A in the 3rd trimester of pregnancy. The overall rise in the blood levels of transferrin under the effect of oestrogens is confirmed. Norms were obtained for the levels of the IgA/T ratio which drops progressively as pregnancy progresses and with oral contraception. It has been found that in the last two trimesters the upper limit of the normal is 1 for the IgA/T ratio as compared with 1.9 in the general population. The study of these constants is of value when screening for maternal alcoholism and as a consequence for the prevention of the fetal alcohol syndrome.


Rat brain ribonucleases.

Rat brain ribonucleases. Rat brain ribonucleases (RNases) were studied. Three types of RNases with maximum activities at pH 5.0, 7.2 and 9.5 were found. The activity of the pH 7.2 enzyme can be detected only by avoiding the interference of a very active inhibitor with p-chlor-mercuri-benzoic acid (PCMB). The effect of bivalent cations (Ca2+, Mg2+), Na+ and ethylenediamine tetraacetic acid (EDTA) was investigated. The activities studied showed a different subcellular distribution. Changes in RNase activity during postnatal rat brain development were studied. The pH 7.2 and 9.5 enzymes have a similar behavior increasing up to the 15th-20th day and remaining constant thereafter. The pH 5.0 enzyme remains constant from the 5th to the 20th day, decreasing thereafter.


Yale Conference on Behavioral Medicine: a proposed definition and statement of goals. The Yale Conference on Behavioral Medicine brought a diverse group of behavioral and biomedical scientists together for the purpose of arriving at an interdisciplinary yet consensual definition, statement of goals, and set of recommendations regarding the emerging field of behavioral medicine. It was proposed that behavioral medicine be defined as "the field concerned with the development of behavioral science knowledge and techniques relevant to the understanding of physical health and illness and the application of this knowledge and techniques to prevention, diagnosis, treatment, and rehabilitation. psychosis, neurosis, and substance abuse are included only insofar as they contribute to physical disorders as an end point." The rational behind this definition and proposals for future developments in the field are discussed.


Studies on experimental growth retardation in sheep. The effect of removal of a endometrial caruncles on fetal size and metabolism. Experimental intrauterine growth retardation was studied in sheep. Endometrial caruncles (anlagen of maternal cotyledon) were removed before pregnancy and at a second operation, catheters were implanted into the ewe and fetus at 105-135 days of pregnancy. Three groups of fetuses were defined: low birthweight-for-dates (small-caruncle), normal birthweight-for-dates (normal-sized-caruncle) from ewes which had endometrial caruncles removed and the controls. The mean placental weights in these groups were 139 plus or minus 5 g, 283 plus or minus 46 g, 334 plus or minus 22 g respectively. The brains, kidneys and adrenals of the small-caruncle-fetuses were significantly greater in proportion to body weight than in the controls and the appearance of ossification centres was delayed. Arterial oxygen tension was lower and packed cell volume higher in the small-caruncle-fetuses (PaO2 15 plus or minus 0.6 mmHg; packed cell volume 37.3 plus or minus 1.6%) and normal sized caruncles (PaO2 20.7 plus or minus 1.2 mmHg; packed cell volume 35.2 plus or minus 0.7%) than in the controls (PaO2 23.2 plus or minus 0.7 mmHg; packed cell volume 29.8 plus or minus 0.7%). Plasma concentrations of glucose (0.65 plus or minus 0.12 micromol/ml), lactate (0.9 plus or minus 0.1 micromol/ml) and pyruvate (0.08 plus or minus 0.025 micromol/ml) were lower in small-caruncle fetuses than in the control fetuses (glucose 1.05 plus or minus 0.06 micromol/ml, lactate 1.83 plus or minus 0.7 micromol/ml, pyruvate 0.21 plus or minus 0.06 micromol/ml). The corresponding values for the normal-sized-caruncle fetuses were glucose 0.71 plus or minus 0.12, lactate 1.18 plus or minus 0.7 and pyruvate 0.12 plus or minus 0.03 micromol/ml. The plasma concentration of alanine in the small-caruncle-fetuses (0.25 plus or minus 0.09 micromol/ml) was higher than in the normal-sized-caruncle (0.073 plus or minus 0.009 micromol/ml) or control fetuses (0.12 plus or minus 0.013 micromol/ml). The results indicate that fetal growth retardation due to restriction of placental growth after removal of endometrial caruncles is associated with chronic hypoxaemia, polycythaemia and hypoglycaemia. The restriction of nutrient supply probably accounts for the altered pattern of fetal growth but the relative importance of the changes observed remains uncertain.


A systematic error in the determination of blood PCO2. An analysis was made of the effect of sodium heparin solution on the determination of arterial blood PCO2, pH, and bicarbonate. Dilution of blood by heparin solution did not affect the pH of blood samples significantly but reduced the measured PCO2 and th calculated bicarbonate and base excess in direct proportion to the amount of dilution. Used in the usual quantities as an anticoagulant, lubricant, and filler of needle and syringe dead space, heparin solution caused a 17 per cent or greater reduction in PCO2 and bicarbonate in approximately 17 per cent of the arterial blood samples currently received for analysis in a university hospital laboratory. Such unrecognized errors could cause misinterpretations of acid-base status. Completely filling the sampling syringe with blood and using a small-gauge needle will reduce the error.


Effect of protein on titrimetry of bicarbonate, titratable acid, and ammonium in urine. Measurement of bicarbonate, titratable acid (HTA+) and ammonium in the urine can facilitate clinical evaluation of acid-base status. Sequential measurement of these three components by titrimetric techniques is well established, but possible interference by proteinuria has not been examined. We report the influence on these analyses of albumin and globulin, two proteins commonly observed in urine in renal disease states. The presence of these proteins in urine affects the measurement of NH4+, and to a less extent that of HTA+. The magnitude of the effect depends on the concentration and the kind of protein present. Proteins do not influence the measurement of HCO3-.


Creatine kinase in serum: 6. Inhibition by endogenous polyvalent cations, and effect of chelators on the activity and stability of some assay components. We studied the effects of some chelators on creatine kinase activity. Creatine kinase is competitively inhibited by endogenous polyvalent cations (e.g., calcium, Ki = 4.5 mmol/L); this can be reversed by adding chelators to the reagent, resulting in a mean increase in activity of 1.14-fold at 25 degrees C and 1.18-fold at 30 and 37 degrees C. Adding chelators, 5 and 10 mmol/L, to serum stored at 37, 30, 25, 4, and -20 degrees C increased isoenzyme stability in some cases, but under certain conditions decreased it, especially at higher chelator concentrations, and more so for EGTA than for EDTA. Blood sampling into tubes prepared with chelators and storage of plasma has no advantage over serum stored in the presence of chelators. The most striking effect of chelators is their protective effect on thiols in the creatine kinase reagent. In the presence of EDTA, 2 mmol/L, the reagent is stable for at least a day at 25 degrees C or a week at 4 degrees C. The poor stability of glucose-6-phosphate dehydrogenase, which is nearly independent of chelators, is the limiting factor for reagent containing EDTA. Bis-Tris, a buffer recently recommended for assay of creatine kinase activity, is a weak chelator. Imidazole acetate buffer combined with EDTA yields activities identical to those found with Bis-Tris at assay temperatures of either 25 or 30 degrees C.


Fluorescent derivatives of nucleotides. Metal ion interactions and pH dependency. The fluorescence parameters of ethenoadenosine derivatives are influenced by metal cations and pH, as summarized here. The pH profile of ethenoadenosine determined by fluorescence intensity gives a normal titration curve and is not affected by ionic strength. In contrast, the pH titration curves of etheno-ATP, etheno ADP, and etheno AMP depend upon ionic strength. At high ionic strength normal curves are obtained, whereas at low ionic strength anomalies are obtained; this suggests that the phosphates can interact with the ring, possibly by hydrogen binding to the ring nitrogens. The room temperature fluorescence of ethenoadenosine occurs from the base form, although excitation of either the acid or base forms can contribute to the emission. This result can be explained if the excited state pK is lower than the ground state pK, and if deprotonation occurs within the time scale of the excited state. At low pH values the fluorescence lifetime of the base form is dependent upon the buffer concentration, indicating that the reverse reaction, protonation, occurs. The affinity constants for the binding of metals to the ethenoadenosine phosphates resemble those for the corresponding adenosine phosphates. Ni(II) and Co(II) are more effective than Mn(II) in quenching the fluorescence of ethenoadenosine phosphates; this result is predicted by Förster's theory for energy transfer based upon the overlap between donor emission spectrum and acceptor absorption spectrum. The diamagnetic ions Mg(II), Ca(II), and Zn(II) do not appear to affect the fluorescence of the ethenoadenosine phosphates directly, but rather to affect the conformation of the molecule, thereby affecting the quantum yield.


The quantum yield of flash-induced proton release by bacteriorhodopsin-containing membrane fragments. The quantum yield of proton release by bacteriorhodopsin was measured from volume changes after excitation of purple membrane fragments by short flashes. At low ionic strengths, about 0.25 mol of protons is released per einstein absorbed. This agrees well with quantum yields reported recently for the conversion of bacteriorhodopsin into a metastable state (M) that absorbs near 412 nm. However, the quantum yield of proton release increases gradually with increasing ionic strength; it plateaus with a value of 0.43 +/- 0.03 at ionic strengths above 200 mM. Changing the ionic strength has no detectable effect on the quantum yield of formation of the M spectral state. It thus appears that as many as two protons can be released and rebound in each photochemical cycle at high ionic strengths. The quantum yield of proton release is essentially independent of pH over the range 6.0-8.75. The quantum yield decreases with increasing flash strength, apparently due to photoreversal of the initial photochemical reaction.


Bacteriorhodopsin in model membranes. A new component of the displacement photocurrent in the microsecond time scale. A quasi-short-circuit (tunable voltage clamp) measurement method with microsecond time resolution was applied to a bacteriorhodopsin model membrane formed by a novel interfacial technique. A new component (B1) of the displacement photocurrent was recorded: it has no detectable latency at an instrumental time constant of 1.5 museconds, and persists at 5 degrees C. In addition, a slower component (B2) of opposite polarity inhibited by low temperature (5 degrees C) and low pH (pH = 3.0) was recorded. The technique is very sensitive for the study of fast capacitative photoresponses in model membranes, and allows the detection of charge displacements in bacteriorhodopsin associated with distinct stages of the photochemical transformation.


Dynamics of pH-induced spectral changes in bacteriorhodopsin. The kinetics of the spectral shift induced in bacteriorhodopsin by low pH are investigated by using the rapid-mixing, stopped-flow technique. The generation of the acid form of the chromophore (A605) occurs in two distinct steps: a fast process (t1/2I = 21 +/- 4 ms) is followed by a much slower reaction (t1/2II = 6 +/- 2 s). The observations are interpreted in terms of neutralization of an acid group in the neighborhood of the retinyl chromophore, the double-staged kinetics being attributed to cooperative effects between chromophores. The method may serve as a tool for studying the kinetics of proton migration across the purple membrane.


K+ conductance modified by a titratable group accessible to protons from the intracellular side of the squid axon membrane. In the range of pH examined (5.2-10), variations of internal pH from high to low values result in a reversible decrease of the conductance of the open K channels, without significantly affecting the kinetics parameters. A linear plot of the conductance versus internal pH suggests the existence of a titratable group that has an apparent pKa of about 6.9, and that is accessible to protons only from the intracellular side of the membrane.


Regulation of nerve terminal calcium channel selectivity by a weak acid site. The effects of low pH, and of alkaline earth cations, were examined on calcium uptake by pinched-off nerve terminals (synaptosomes). This uptake appears to be mediated by voltage-sensitive Ca channels (J. Physiol. 247:617, 1975). Ca uptake was measured in low (5 mM) or high (77 mM) potassium media. The extra uptake promoted by depolarizing (K-rich) media was almost maximal at pH 7.5, and decreased as the pH was lowered. Data relating depolarization-induced 45Ca uptake to pH fit a titration curve with a pKa approximately 6. Experiments in which Ca concentration and pH were both varied indicated that Ca2+ and H+ compete for a common binding site. Inhibition of depolarization-induced 45Ca uptake by the alkaline earth cations was studied to determine the apparent binding sequence for these cations in the Ca channels: Ca greater than Sr greater than Ba greater than Mg. This sequence resembles that observed for block of Ca channels in other preparations. The apparent binding sequence of the alkaline earth cations and the apparent pKa (approximately 6) of the Ca-binding site indicate that the Ca channel is a "high field strength" system. Protonation of a Ca channel binding site could explain the inhibitory effect of low pH on Ca-dependent neurotransmitter release (cf. Del Castillo et al., J. Cell. Comp. Physiol. 59:35, 1962).


Nuclear magnetic resonance determinations of permeation coefficients for maleic acid in phospholipid vesicles. Lipid bilayer permeation coefficients for the neutral maleic acid molecule and the maleate monoanion have been determined by proton magnetic resonance techniques. Phosphatiydylcholine-cholesterol (2:1) unilamellar vesicles were prepared having an initial maleate anion concentration gradient stabilized by coupling to an impermeant potassium counterion. The coupling was released by addition of valinomycin, and the time evolution of external pH, internal pH, and maleate concentration followed using nuclear magnetic resonance areas and chemical shifts. Transport rate equations were numerically integrated to fit the date, yielding best fit permeation coefficients of 4 X 10(-9) and 4 X 10(-5) cm/s for maleate monoanion and maleic acid, respectively.


Viscoelastic behavior of mammalian DNA.

Viscoelastic behavior of mammalian DNA. The viscoelastic behavior of rat 9L cellular DNA was studied as a function of the detergent used for lysis, the pH and duration of lysis, and gamma ray dose. For nondenaturing lysis conditions, a model of the DNA was proposed to account for the effects of these agents on the viscoelastic retardation time. It was concluded that these agents affect the hydrodynamic radius of the DNA rather than its molecular weight. For denaturing lysis conditions, molecular weights calculated from the relaxation time were consistent with those calculated from alkaline sucrose sedimentation profiles.


Suppression of graft-versus-host reaction by preincubation of the graft with an antitumor protein, neocarzinostatin. The immunosuppressive activity of neocarzinostatin, an antitumor antibiotic with a high molecular weight, was demonstrated, as measured by the local and systemic graft-versus-host reaction in rats. The preincubation of parental strain lymphocytes with doses more than 1 micrograms/ml of neocarzinostatin for at least 10 min at 37 degrees C resulted in the marked suppression of the popliteal lymph node enlargement, when injected subcutaneously into the hind footpads of F1 hybrid rats. The suppressive effect was temperature-dependent and irreversible, because the neocarzinostatin (1 microgram/ml) pretreatment of parental lymphocytes at 4 degrees C was not effective in the suppressive activity, and the reincubation of cells after the removal of the drug from the culture did not result in demonstrable changes in the degree of suppression. The neocarzinostatin pretreatment does not result in visible changes in the oxygen consumption rate and viability of cells in vitro or in the distribution pattern within a regional lymph node, indicating that treated cells may retain the same viability and migration capacity in vivo as do untreated cells. Furthermore, the drug pretreatment resulted in the complete prevention of runting syndrome, when treated parental lymphocytes were injected intravenously into F1 hybrid rats.


Lymphocytotoxicity and immunosuppression by organotin compounds. Suppression of graft-versus-host reactivity, blast transformation, and E-rosette formation by di-n-butyltindichloride and di-n-octyltindichloride. Di-n-butyltindichloride (DBTC) and di-n-octyltindichloride (DOTC) represent a new group of organometallic compounds with antilymphocytic properties. In rats they induce lymphocyte depletion in thymus and thymus-dependent areas of spleen and peripheral lymph nodes without signs of myelotoxicity or a generalized toxicity. The number and viability of cells isolated from thymus and peripheral lymphoid organs was severely decreased, whereas the number and viability of bone marrow cells was not reduced. Immunosuppressive properties of DBTC and DOTC are indicated, in this study, by a severe decrease of the graft-versus-host response and the response to the T-cell mitogens phytohemagglutinin and concanavalin A. The T-cell selectivity of these compounds is discussed. In vitro DBTC and DTOC are extremely cytotoxic. Blast transformation of human as well as rat thymocytes was already inhibited at concentrations as low as 0.02 micrograms DBTC (or 0.1 micrograms DOTC) ml medium. Also the E-rosette formation was inhibited at very low drug levels. The similarity of effects upon rat and human lymphocytes suggests that DBTC and DOTC acts in the same manner in rat and man and offers the possibility of a therapeutic use of these compounds.


Extracellular pH selectively modulates recovery from sodium inactivation in frog myelinated nerve. The Hodgkin-Huxley kinetic parameters, alpha h and beta h, which govern the rate of recovery from and development of sodium channel inactivation, respectively, have been measured as a function of membrane potential and external pH using a three-pulse protocol. alpha h but not beta h is substantially accelerated by reducing external pH from 7.4 to 6.4. The alpha h vs. voltage curve appears to be selectively shifted in the depolarizing direction by approximately 12 mV for this pH change, giving an apparent, h infinity curve shift of approximately 6 mV in the same direction (less inactivation).


Persistence of improved exercise tolerance and degree of revascularization after coronary bypass surgery. A prospective randomized study. Exercise tolerance was repeatedly determined over a 2-year period in a series of 100 patients with coronary heart disease randomly allocated for medical therapy and coronary bypass surgery. The surgical group had a consistently better exercise tolerance than the medical group during the whole follow-up. Completeness of the revascularization, assessed by repeated graft and native vessel angiography, resulted in a marked improvement whereas incompletely revascularized patients exhibited only a marginal improvement which, nevertheless, to some degree exceeded the result of medical management alone. It is concluded that coronary bypass surgery and medical therapy, when indicated, result in markedly better exercise tolerance than medical management alone. This improvement persists up two years after the operation and is largely dependent on the completeness of the revascularization.


Detection and properties of TSH-binding inhibitor immunoglobulins in patients with Graves' disease and Hashimoto's thyroiditis. TSH-binding inhibitor immunoglobulins (TBII) have been detected in patients with Graves' disease and Hashimoto's thyroiditis by using the radioreceptor assay of TSH. In untreated Graves' patients, TBII levels correlated well with thyroidal 99mTc uptake at 30 min and the grade of epithelial hyperplasia of thyroid follicles. There were many Graves' patients whose sera contained high TBII levels but no detectable bioassayable thyroid-stimulating activity (LATS), and in these patients, close correlation was observed between serum levels of TBII and bioassayable LATS-protector activity. TBII were detectable in 2 (10%) of 20 patients with Hashimoto's thyroiditis, both of whom were clinically hypothyroid. The serum or IgG fraction from one of them, however, did not contain any significant LATS, LATS-protector, or human thyroid adenylate cyclase-stimulating activity and caused inhibition of adenylate cyclase stimulation by TSH. In that patient, TBII may be acting to block TSH binding to TSH receptors, thus causing TSH unresponsiveness and hypothyroidism.


Aldose reductase in diabetic complications of the eye. Aldose reductase (AR) appears to initiate the cataractous process in galactosemic and diabetic animals. Sugars in excess are converted to polyols by lens AR. In sugar cataracts, polyols accumulate to levels substantial enough to cause a hypertonicity leading to lens fiber swelling. All other changes appear secondary to polyol accumulation and lens swelling. The development of sugar cataracts can be duplicated in organ culture. In culture, the various changes that occur were minimized or did not occur when inhibitors of AR were included in the medium. Moreover, AR inhibitors were shown to effectively delay the onset of sugar cataract development in animals. A defect in the corneal epithelium of diabetics became apparent in vitrectomy. One manifestation of this problem was the delay in the reepithelialization of denuded corneas. In examining this problem experimentally, the epithelium was removed from the corneas of diabetic and normal rats. The regeneration of epithelium in corneas of diabetic rats required a longer period than in the normal. The possibility that AR, active in the epithelium, was involved in this phenomenon was investigated. The corneal epithelium was removed from both eyes of a diabetic rat. One eye was treated topically with the AR inhibitor CP-45,634 while the other served as control. The eye treated with CP-45,635 regenerated epithelium much more quickly than the untreated eye. Other AR inhibitors had similar beneficial effects.


Recovery of the gastric mucosa from bile-induced injury. Bile salts are known to injure the gastric mucosa and may be involved in the pathogenesis of gastric stress ulceration, a feared sequela of shock. Although the induction of bile injury to the stomach has been extensively studied, recovery from such an injury has not. In the studies reported here the period after removal of bile was examined to see whether the mucosa could return to pre-bile-exposure status under various circumstances. Recovery from a bile salt-induced injury was studied in three mongrel dogs with Heidenhain pouches. Net acid back diffusion (NABD) was measured with a recirculating system incorporating a pH stat and was measured with a recirculating system incorporating a pH stat and autoburette, and transmucosal electrical potential difference (PD) was measured by usual methods. The effects of exposure to 5 mM taurodeoxycholate at pH 2 for 30 minutes were studied. This exposure increased net acid back diffusion from 23 +/- 5 to 184 +/- 16 micro Eq/10 min (P less than 0.01) and reduced PD from 56 +/- 2 to 18 +/- 2 mV (P less than 0.01). After injury the pouches were copiously irrigated with saline to remove bile, and then recovery was observed as the pouches were perfused with pH 1, 2, or 7 HCl solution made isotonic with NaCl. The recovery values were compared with the same dog's preinjury values at the same pH perfusion. The data showed that at pH 2 and pH 7, PD and NABD returned to preinjury levels within 90 minutes after bile injury. At pH 1, however, recovery did not occur. The pH dependence of recovery from bile-induced injury may have relevance to patients with episodic reflux of bile into the stomach.


Viability of some Salmonella strains in Algerian eggs. Whole-egg and egg-white were inoculated with S. typhi murium, S. dublin, S. oranienburg and S. oslo separately and were stored at - 1 degrees C, + 4 degrees C and 26-28 degrees C. pH value was determined every day as well as the survival of the different inoculated Salmonella strains till the end of the experiment. Stored at - 1 degrees C in case of whole-egg, egg-yolk and egg-white, pH values were 7.0, 6.0 and 7.0 while the maximum values were 8.5, 8.0 and 9.0 respectively. The shortest survival period for Salmonella strains was 101 days in whole-egg, 103 days in egg-yolk and 36 days in egg-white, while the longest period was 114, 123 and 59 days respectively. The minimum pH values of whole-egg, egg-yolk and egg-white stored at + 4 degrees C were 6.5, 6.0 and 7.0 while the maximum values were 8.5, 8.5 and 10.0 respectively, 55, 66 and 30 days were the shortest survival periods for Salmonella strains inoculated in whole-egg, egg-yolk and egg-white while the maximum periods were 81, 80 and 67 days. The minimum pH values of whole-egg, egg-yolk and egg-white held at room-temperature, were 6.0, 6.0 and 7.5 while the maxima were 8.5, 8.5 and 10.0 respectively. The minimum survival period of the Salmonella strains inoculated in whole-egg, egg-yolk and egg-white was 52, 41 and 21 days, while the maximum periods were 63, 52 and 37 days respectively.


5,7-dihydroxytryptamine lesions of dorsal and median raphé nuclei and performance in the social interaction test of anxiety and in a home-cage aggression test. Micro-injections of the neurotoxin 5,7-dihydroxytryptamine into the dorsal raphe nucleus produced a behavioural profile in the social interaction test of anxiety similar to that seen in rats treated chronically with benzodiazepines. Neurotoxin injections into the median raphé nucleus did not produce a profile significantly different from that of the controls. In the control rats and in the rats with lesions of the median raphé nucleus, ACTH1-24 (corticotrophin) significantly reduced active social interactions, whereas it was without effect on the rats with lesions of the dorsal raphé nucleus. In the home-cage intruder test, the median raphé-lesioned rats submitted less to the intruder and stood and jumped on him more often than did the controls. The dorsal raphé-lesioned rats showed significantly fewer interactions of all kinds, compared with control rats when an intruder was placed in their home cages.


beta-Adrenoceptor blockers in the treatment of hypertension. Clinical drug trials have shown that pindolol, timolol and sotalol are effective hypotensive agents in Nigerians with mild to moderate hypertension (standing diastolic blood pressure below 120 mmHg). There was no significant difference between the three beta-blockers with respect to their antihypertensive effect irrespective of differences in their other pharmacological actions. There was also no correlation between the hypotensive effect of the drugs and the initial blood pressure. Timolol at 30 mg daily had an additive effect to the hypotensive action of binazine, a peripheral vasodilator. Increasing the dose of timolol to 60 mg daily did not produce further fall in the blood pressure. Pindolol at a dose of 30 mg per day also had an additive effect to the hypotensive action of the thiazide diuretics, methyldopa and debrisoquine. There was no significant difference between the hypotensive effect of sotalol and that of methyldopa. The three beta-blockers produced no side effects. This was considered to be a distinct advantage over most of the drugs currently available for treatment of hypertension in Nigeria.


The lower esophageal sphincter in gastroesophageal reflux in children. Esophageal function was evaluated in 51 children less than 2 years of age with radiologic evidence of gastroesophageal reflux. Detection of an acid esophageal pH was a sensitive measure of gastroesophageal reflux. Lower esophageal sphincter pressures were greater in reflux patients with respiratory symptoms (18.0 +/- 1.4 mm Hg) than in reflux patients without respiratory symptoms (9.5 +/- 1.0 mm Hg). The intra-abdominal segment of the lower esophageal sphincter was shorter in patients with reflux than in controls (0.51 +/- 0.05 cm vs. 0.75 +/- 0.08 cm). It was also shorter in patients requiring surgical therapy (0.34 +/- 0.05 cm) than in those responding to medical therapy (0.63 +/- 0.07 cm).


Takayasu's arteritis: possible dysgenesic etiology. A 31-year-old female patient with Takayasu's arteritis and vascular abnormalities is presented. Her dominant symptom was intermittent claudication in the left upper extremity. The arch aortogram and selective angiogram showed obstruction of the left subclavian and axillary arteries, and run-off in the proximal brachial artery. An anomalous common trunk for the innominate and left carotid arteries and a markedly narrowed left vertebral artery were found. A direct surgical attempt at improving the blood supply of the left upper extremity was not deemed feasible. A transthoracic cervicodorsal sympathectomy was done without lasting functional results. The relatively frequent association of aortic arch branching abnormalities, also abnormalities of the thoracic, abdominal aorta, of the coronary, visceral arteries with aortic arch syndrome and other arteriopathies had been noted in several reports. It appears that such uncommon vascular malformations are not without influence upon the development of these forms of obstructive arteritis. The mechanism of the obstructive process is unclear. The possible dygenesic etiology of Takayasu's disease is herein postulated.


Effect of the frequency of stimulation on the blocker action of propranolol in isolated rat left atria. The beta-blocking activity of propranolol was studied on the positive inotropic effect of norepinephrine, epinephrine, isoproterenol, dopamine and ethylephrine, in the left atrium driven at different rates. Dose of the antagonist of 10(-8) and 10(-7) M did not block the norepinephrine dose-response curve at 1 and 2.8 Hz, but a shift to the right was observed at 1.6 Hz. Although epinephrine showed a significant increase in its pD2 (p less than 0.001) at the lower stimulation frequency, the blockade increased progressively the higher the rates. On the other hand, propranolol antagonized isoproterenol at all the frequencies tested, in spite of an increase in the maxima at 2.8 Hz. This latter behavior was also true for dopamine and ethylephrine. Both sympathomimetic amines were blocked by propranolol at 1 and 1.6 Hz. The complex effect of propranolol on Ca2+ movements and its effects on cAMP and ATPase seem to be superimposed to the beta-blocking activity. Thus, the various actions on the sympathomimetic amines change according to the agonist considered and the stimulation frequency employed.


[Neurohumoral control of the pineal gland. A model for the study of neuroendocrine integrative processes]. This article discusses the experimental evidence which suggests that the pineal gland and its innervating neurons are useful paradigms for the study of neuroendocrine integrative processes. The obtained results can be summarized as follows: 1) existence of putative receptors for various hormones (estradiol, testosterone, 5 alpha-dihydrotestosterone, progesterone, prolactin) in the mammalian pineal gland; (2) steroid metabolic pattern in the pineal gland resembling other brain areas involved in gonadotrophic regulation; (3) control of pineal estrophilic and androphilic receptors by adrenergic transmitter through beta-adrenergic receptors and at a translational level; (4) denervation supersensitivity of hormone receptors to neurotransmitter; (5) modification of neuronal activity by hormone treatment at ganglionic and preganglionic sites of action; (6) estrophilic binding sites in ganglia; (7) correlation of pineal responsiveness to hormones with activity of sympathetic nerves; (8) modification by hormones of pineal beta-adrenergic mechanisms; (9) dissociation of hormone effects on the pineal gland in those mediated or modulated by changes in afferent neuronal activity and those relatively unaffected by denervation. Collectively these data indicate that neuroendocrine, endocrine-neural and endocrine-endocrine transducing processes occur in the pinealocytes and superior cervical ganglia.


Prospects for accelerating hematopoietic recovery following myelosuppressive therapy by using autologous, cryopreserved hematopoietic stem cells collected solely from the peripheral blood. Cryopreserved, autologous stem cells collected from human marrow have been used to accelerate hematopoietic recovery following intensive radiation or chemotherapy. The harvesting of adequate numbers of bone marrow cells for purposes of hematopoietic reconstitution is a potentially morbid procedure and requires the use of general anesthesia and blood transfusion during anesthesia. The ability to accelerate hematopoietic recovery using cells collected solely from the peripheral blood would obviate the requirement for a general anesthetic and avoid the discomfort experienced by patients following the procedure. Collection of hematopoietic stem cells from the peripheral blood should involve limited morbidity and risk and be well suited to repeated application in individual cases. An increasing body of in vitro and in vivo data in animals and man suggests that this goal may soon be within reach.


Isolation, cryopreservation, and autotransplantation of human stem cells. We have used cryopreserved autologous hematopoietic cells to repopulate bone marrow of patients receiving intensive chemotherapy and radiation. We employed the technique of density step centrifugation to concentrate stem cells from whole bone marrow. The concentrate of cells was frozen in vials using dimethyl sulphoxide (DMSO) as a cryo-protective agent and was stored in the vapor phase of liquid nitrogen. Test vials were thawed and assayed for myeloid stem cells (CFU-C) to determine the potential of the larger marrow aliquot. These test values were compared to the recovery of cells and CFU-C when the marrow was used for autotransplantation. The return of hematopoietic function after autotransplantation was evaluated and correlated with the dosage of cells and CFU-C infused.


Processing of peripheral blood stem cells for transplantation. Canines were studied to determine the efficacy of peripheral blood collection by semicontinuous centrifugation to procure sufficient numbers of hematopoietic cells for marrow reconstitution. CFU-C assays on fresh and cryopreserved peripheral blood cells were compared to bone marrow aspirates. Attempts were made to partially separate hematopoietic cells from immunocompetent cells during buffy coat collection and by incubation with activated cyclophosphamide. Yields from 4 runs of semicontinuous centrifugation averaged 1.7 x 10(5) CFU-C compared to 1.9 x 10(5) CFU-C for standard marrow aspiration. Ratios of approximately 5/1 marrow to peripheral blood mononuclear cells (MNC) were found. Autologous transplantation with 1.6 x 10(4) CFU-C per kg resulted in evidence of marrow reconstitution within two weeks following otherwise lethal chemotherapy. Recovery of CFU-C following cryo-preservation was 82% for marrow and 78% for peripheral blood. Selective depression of MLC activity occurred when peripheral blood MNC were incubated for 30 minutes with activated cyclophosphamide. MLC was inhibited by greater than 90% while 70% of CFU-Cs were retained. It was concluded that peripheral blood may be a practical alternative to marrow for transplantation studies.


Neuroendocrine modulation of calcitonin and parathyroid hormone in man. Recent evidence suggests that of calcitonin (CT) and parathyroid hormone (PTH) is controlled by factors other than the ambient serum calcium concentration. We studied the effects of infusions of four neuroendocrine modulators upon CT and PTH levels: isoproterenol (beta-adrenergic agonist), methoxamine (alpha adrenergic agonist), prostaglandin E2, and somatostatin. Isoproterenol was a consistent secretagogue for both hormones. Maximal CT increments during isoproterenol infusion in normal subjects were 13 +/- 2 pg/ml (mean +/- SEM, n = 6, P less than 0.001; basal, 26 +/- 5). Maximal increments in PTH were 113 +/- 22 pg/ml (P less than 0.01, n = 6; basal, 430 +/- 11). Infusions of methoxamine increased CT by 13 +/- 5 pg/ml (n = 5, P less than 0.05; basal, 43 +/- 13), but had no effect on PTH. The means of the maximal CT increments during isoproterenol (21 +/- 8 pg/ml) and methoxamine infusion (28 +/- 11 pg/ml) were not statistically different from those achieved by acute elevations of serum calcium levels within the physiological range (41 +/- 23 pg/ml). Infusions of somatostatin and prostaglandin E2 had no or only transient effects on basal or stimulated CT or PTH levels. Our data suggest that adrenergic input modulates CT and PTH secretion in humans independently of changes in serum calcium.


In vitro effect of dopamine and pimozide on human chorionic somatomammotropin (hCS) secretion. In human placental explants cultured in vitro, dopamine inhibited human chorionic somatomammotropin (hCS) secretion into the culture media. In the control flasks, the level of hCS secretion was 130.5 +/- 7.8 micrograms/g tissue (n = 6). When 1 mM dopamine was added, hCS levels decreased to 80.2 +/- 11.5 micrograms/g tissue (P less than 0.01). Dopamine (5 and 10 mM) further lowered hCS levels. In contrast, 1 mM pimozide enhanced hCS secretion by 2-fold as compared to control levels (248.2 +/- 44.8 vs. 130.5 +/- 7.8, P less than 0.02). The simultaneous addition of dopamine did not alter the stimulatory effect of pimozide on hCS secretion. In separate experiments, arginine (1 and 5 mM) and somatostatin (1 microgram/ml culture media) did not alter hCS secretion from placental explants. These results suggest that hCS secretion is modulated by dopaminergic receptors.


Zoological specificity of human thyroid-stimulating antibody. The zoological specificity of human thyroid-stimulating antibody (TSAb) that occurs in the blood in Graves' disease was examined by assessing its effect on the thyroid of the dog, guinea pig, calf, and mouse as well as that of man. With all but murine glands, thyroid stimulation was assayed by measuring the increase in the concentration of cAMP in the thyroid slices or fragments after 2 h of incubation in buffer containing TSAb. Effects on the thyroid of the mouse were monitored by the in vivo bioassay for LATS. Sera from 33 patients with Graves' disease were obtained and concentrates of TSAb were prepared by precipitation of IgG with 1.64 M (NH4)2SO4. These all stimulated the human thyroid, 13 were LATS-positive, and they variably affected the tissues of other species; of 27 tested, 14 stimulated the thyroid of the dog, 8 out of 23 stimulated the thyroid of the guinea pig, and 12 out of 16 stimulated the gland of the calf. The more potent the TSAb as assayed with human tissue, the more likely was it to stimulate other species of thyroid; however, frequent exceptions occurred. In a separate analysis of 35 LATS-positive preparations of TSAb, correlation between the responses in the LATS and human thyroid slice assays was statistically significant (P less than 0.001). The data are compatible with the view that stimulation by TSAb of nonhuman thyroids, including the murine as in the LATS bioassay, reflects cross-reaction of this immunoglobulin with an antigen that has sufficient similarity to the human molecule to be recognized by the human antibody.


Effect of the dopamine agonist, lergotrile mesylate, on circulating anterior pituitary hormones in man. The effects of the ergoline derivative, lergotrile mesylate, on the serum levels of PRL, GH, TSH, LH, FSH, cortisol, and blood sugar were studied in six normal males. The effects of lergotrile mesylate on the serum levels of GH and PRL were also studied in eight patients with acromegaly and in two with idiopathic hyperprolactinemia. In the normal subjects, 2 mg oral lergotrile lowered basal PRL levels after 90 min and markedly impaired the PRL response to TRH (200 micrograms iv); the mean peak value +/- SE was 8.3 +/- 1.1 micrograms/liter, compared to the control value of 66.6 /+- 11.3 micrograms/liter. Lergotrile raised serum GH levels in five of the six subjects to peaks of 8-49 micrograms/liter, compared to 2-8 micrograms/liter after placebo. In three subjects, the GH response to lergotrile was attenuated by the prior administration of the dopamine antagonist, metoclopramide (10 mg orally). Lergotrile had no effect on FSH and LH levels under basal conditions or after the gonadotrophin-releasing hormone (GnRH; 100 micrograms iv). Circulating TSH levels were unaltered basally but impaired after TRH. Blood sugar levels were unaltered; serum cortisol was elevated in five of six subjects; there was a brief depression of diastolic blood pressure, but no change in pulse rate. The side effects after lergotrile were variable, with drowsiness as a consistent feature. These actions are similar to those of bromocriptine (an ergot derivative treatment of hyperprolactinemia and acromegaly, to suppress PRL and GH secretion, and in parkinsonism. Therefore, it may be expected that lergotrile could fulfill these clinical uses; however, in the studies comparing the effects of single oral doses of lergotrile (2 mg) and bromocriptine (2.5 mg) on GH and PRL secretion in patients with acromegaly and hyperprolactinemia, lergotrile in the dose used has been found to have an earlier onset and shorter duration of action.


Failure of alpha-methyltyrosine to inhibit peripheral triiodothyronine formation. To determine if the adrenergic nervous system, and specifically tyrosine hydroxylase, plays a role in the extrathyroidal conversion of T4 to T3, normal male volunteers were treated with T4 and subsequently with T4 and alpha-methyl-p-tyrosine (alpha-MPT), an inhibitor of tyrosine hydroxylase, for 2 weeks. The mean serum T4 and T3 concentrations increased during T4 administration and remained at the same levels during combined T4 and alpha-MPT administration. Urinary vanillylmandelic acid excretion declined significantly during alpha-MPT administration. These results do not support the hypothesis that tyrosine hydroxylase is involved in extrathyroidal T3 production.


GH secretion in two siblings with Laron's dwarfism: the effects of glucose, arginine, somatostatin, and bromocryptine. The secretion of GH in two siblings with clinical dwarfism and high GH plasma levels (the mean of several basal values; 233.83 ng/ml in patient A and 178.16 in patient B has been studied with several dynamic tests. An arginine infusion increased GH levels in both cases (+193.55% for A, +140.27% for B). No significant modifications were obtained with oral glucose tolerance test +18.70% for A, +24.32% for B). A bolus of somatostatin almost completely prevented the rise in GH levels in response to arginine. Pretreatment with bromocryptine clearly increased basal GH plasma levels (A, +58.66%; B, +56.03%) and the response to arginine. As in the case of a normal hypothalamus, the hypothalamus of Laron's syndrome responds to arginine and bromocryptine, with GH elevations. Somatostatin suppresses GH levels. A lack of response to glucose can be considered as a nonspecific effect of the very low biological activity of the stimulus in a hyperstimulated hypothalamus. We suggest that GH secretion by the hypothalmo pituitary system in Laron's syndrome is normal, and that GH hyperproduction may be due to a generalized defect in GH receptors or to the low levels of somatomedin.


Inhibitory effect of guanfacine, a central alpha-adrenoceptor agonist, on prolactin secretion stimulated by insulin-induced hypoglycemia. In six normal male volunteers oral administration of an alpha-receptor agonist, guanfacine (1 mg/q.i.d. for 4 days), had no effect on PRL release induced by 5 mg metoclopramide iv. The same treatment with quanfacine in six other normal subjects significantly reduced PRL secretion stimulated by insulin-induced hypoglycemia (P less than 0.05). These results suggest that an adrenergic pathway, hypothalamic or extrahypothalamic, might be involved in the inhibitory control of PRL secretion.


A comparison of cold and acid activation of big renin and of inactive renin in normal plasma. Normal human plasma contains "inactive renin," whose ability to generate angiotensin I increases after exposure to pH 3.3. Big renin is a partially inactive enzyme of larger molecular weight, which is also activated at pH 3.3, and is found of pregnant women, and in amniotic fluid, but not in normal plasma. We have compared the effects of acid exposure and storage at 4 and -4 C on normal plasma and plasma containing big renin. The concentration of inactive renin in normal plasma was approximately equal to that of normal active renin, and its activity increased slowly on prolonged standing at -4 but not 4 C. In contrast, the activity of big renin increased by 50% as early as 1-3 days at 4 C and increased even more quickly at -4 C. Acid treatment of plasma containing big renin caused 4-10 times greater increase in active renin than similar treatment of normal plasma. During gel filtration, both cold-activated and previously acidified big renin coeluted with unactivated big renin. These data indicate that big renin is highly susceptible to cold or acid activation and that such activation of big renin does not result in a detectable decrease in its molecular weight of 60,000 daltons. Furthermore, acid and cold seem to activate the same pool of inactive renin in normal plasma. Although both normal and big renin are stable for long periods below -20 C, a serious overestimate of plasma renin activity can occur if plasma is stored just above its freezing point before assay.


Effect of dopamine agonist (Lergotrile mesylate) therapy on twenty-four hour secretion of prolactin in treated Parkinson's disease. Plasma PRL was measured at 20-min intervals in six patients with Parkinson's disease under various treatment protocols. In addition, 24-h mean GH levels were measured. The results of these studies showed that two untreated patients with Parkinson's disease had normal 24-h mean PRL levels with the normal increase during sleep. During chronic treatment with L-dopa-carbidopa (Sinemet), the 24-h PRL level was 12.8 +/- 4.9 ng/ml (mean +/- SD) and there was persistence of augmented PRL secretion during sleep. The 24-h mean GH level ranged from 1.5-4.4 ng/ml, with a mean of 2.5 ng/ml. The addition of a dopamine agonist (Lergotrile mesylate) resulted in a significant (P less than 0.01) suppression of the 24-h mean PRL levels and abolition of the normal sleep augmentation after 2 weeks of therapy. This suppression was maintained in one patient who was restudied 4 months after the addition of dopamine agonist therapy to L-dopa-carbidopa. The 24-h mean GH levels did not change significantly after the addition of the dopamine agonist when compared to L-dopa-carbidopa alone. These results suggest a dichotomy between the PRL and GH responses to combined L-dopa-carbidopa and dopamine agonist therapy. In addition, the preservation of normal PRL regulation in the two untreated patients with Parkinson's disease suggests that dopaminergic neurons are not universally affected in this disorder.


Thyrotropin receptors in normal and pathological human thyroid tissues. The properties of TSH receptors in normal and pathological human thyroid tissues were studied. Highly purified bovine TSH after lactoperoxidase iodination retained full biological activity, as assessed by radioreceptor assay. Binding of bovine [125I]TSH to 1000 x g pellets of human thyroid homogenate was specific with respect to hormone and tissue. Total binding amounted to 50-60% of total radioactivity using 10 mg (wet weight) normal thyroid tissue. Nonspecific binding was only 6% of total radioactivity. Normal thyroid tissue contained two orders of binding sites, which were shown to be independent of each other by Hill plot analysis. The high affinity sites [equilibrium dissociation constant (Kd), 0.015-0.16 x 10-9 M] were present in concentrations of 1.05-9.30 pmol/mg protein and concentrations of low affinity sites (Kd, 1.2-2.4 x 10-9 M) were 35.9-213 pmol/mg. In all pathological thyroid tissue studied, two orders of binding sites were found with dissociation constants similar to those of normal tissues, but the number of binding sites was markedly reduced. Both orders of binding sites in solitary "cold" adenomas and only the low affinity sites in thyroid tissue from patients with Graves' disease were significantly reduced in number (P less than 0.01). There was a questionable decrease in high affinity sites in the Graves' tissue (P = 0.05). We have found the definite presence but a decreased number of binding sites in both orders of receptors in papillary and follicular carcinomas. There were few or no binding sites in medullary carcinoma.


Failure of somatostatin to affect human chorionic somatomammotropin and human chorionic gonadotropin secretion in vitro. The effect of somatostatin (SRIF) on human chorionic somatomammotropin (hCS) secretion was studied in human placental explants cultured in vitro. In the experimental flasks, SRIF was added in a concentration of 10, 100, and 1000 ng/ml media; hCS levels measured by RIA were not different from those found in the control flasks. In separate experiments, we investigated the action of SRIF on hCG secretion by a human malignant choriocarcinoma cell line maintained in tissue culture. SRIF (1000 ng/ml) did not inhibit basal or dibutyryl cAMP-induced stimulation of hCG secretion. These results suggest that somatostatin does not suppress hCS or hCG release in vitro from normal or malignant trophoblast, respectively.


Persistent stimulatory effect of glucagon on glucose production despite downregulation. The rise and subsequent return to basal of glucose production (Ra) during a constant glucagon infusion ("downregulation") has suggested to some workers that glucagon's effects are evanescent. To examine whether glucagon displays persistent biological activity even after downregulation, 6 healthy males received an 8 hour infusion of somatostatin and glucagon, with 3H-3-glucose to measure glucose turnover. Ra rose from 2.8 +/- 0.3 to 4.2 +/- 0.3 mg/kg . min at 90 minutes, returned to basal levels at 150 minutes, and remained at this level for the ensuing 330 minutes. Six additional subjects received an 8 hour somatostatin infusion, with glucagon administered concomitantly for the first 5 hours. Glucagon withdrawal at 5 hours produced an immediate decline in Ra from 1.8 +/- 0.2 to 0.9 +/- 0.2 mg/kg . min. Thus, even after downregulation the maintenance of basal Ra is dependent on circulating glucagon.


Somatostatin inhibits thyroid hormone secretion induced by exogenous TSH in man. The effect of somatostatin on thyroid hormone secretion stimulated by TSH in man was studied. The injection of TSH into a thyroid artery during surgery was followed by an increase in the serum concentration of iodothyronines in the corresponding thyroid vein. This increase was significantly lower (p less than 0.02) when somatostatin was given as a bolus injection into a peripheral vein 5 min prior to the administration of TSH, and followed by a continuous infusion for 60 min. Since somatostatin-like immunoreactivity has been localized to some cells of the thyroid gland being in a parafollicular location, it is suggested that somatostatin may be an intrathyroidal regulator of thyroid activity in man.


Serum protein inhibition of thyrotropin binding to human thyroid tissue. We used a modification of the TSH radioreceptor assay to detect TSH-binding inhibition (TBI) activity in serum and serum fractions from normal subjects and patients with Graves' disease. TBI activity is present in normal IgG prepared by DEAE-Sephadex chromatography and in normal globulins prepared by precipitation at 1.6 M ammonium sulfate. Other normal serum proteins also had TBI activity when large concentrations were tested. Gel filtration chromatography and powder block electrophoresis were used to prepare fractions of normal and Graves' disease sera. In these fractions from normal serum. TBI activity was found in both gamma-globulin and alpha-globulin-albumin fractions electrophoretically and in both 7S and 4S peaks from gel filtration. TBI activity from Graves' disease patients' sera was similarly distributed, but relatively more TBI accompanied the electrophoretic gamma-globulins. Sepharose Protein-A and anti-IgG were used as immunoabsorbents to isolate and purify IgG from normal and Graves' disease sera. TBI activity in IgG was proportional to the IgG concentration, indicating that the TBI which migrates as a gamma-globulin electrophoretically is an IgG and thus may possibly be an antibody. Inhibitory activity found in normal serum globulins and the non-IgG fractions of both normal and abnormal sera seriously interferes with attempts to use the TSH radioreceptor assay to study the hypothesized anti-TSH, receptor antibody in the serum of patients with Graves' disease.


Disorders of phagocyte function: biochemical aspects. Intensive laboratory investigation of patients with recurrent infections, and with infections with microbial species not usually considered to be pathogenic, have led to the identification of several defects in granulocyte function. The two functions of granulocytes which have received most attention in the past decade have been locomotion (especially response to chemotactic stimulation) and microbicidal activity. Defective granulocyte chemotaxis has been demonstrated in patients with clinical manifestations suggesting abnormalities related to vasoactive amines, i.e., patients with eczema and extreme IgE hyperimmunoglobulinemia. The depressed granulocyte chemotactic responsiveness found in these patients can be reproduced in vitro when histamine and beta adrenergic agents are incubated with control granulocytes. Since these compounds have been shown to increase levels of intracellular cyclic AMP in other cells, there appears to be an association between cyclic nucleotide metabolism and regulation of granulocyte locomotion. Defective granulocyte microbicidal activity is found in patients with chronic granulomatous disease and it has been shown that there is little increase in oxidative metabolism during phagocytosis by these cells. Methods for quantitating the oxidative metabolism of granulocytes and monocytes include oxygen uptake, reduction of nitroblue tetrazolium, formate oxidation, and chemiluminescence response during phagocytosis. Since products of oxygen metabolism, i.e., hydrogen peroxide, superoxide or singlet oxygen do not accumulate in granulocyte phagocytic vacuoles, intracellular microbes are not killed (except bacterial species that produce hydrogen peroxide). The biochemical basis for defective oxidative metabolism in granulocytes from patients with chronic granulomatous disease appears to be associated with abnormal nucleotide oxidase activity.


Failure of total calcium corrected for protein, albumin, and pH to correctly assess free calcium status. The clinical effectiveness of published algorithms in correcting serum total calcium (CaT) for the effects of protein, albumin, and pH was tested. Corrected calcium (CaC) values obtained by 13 of these methods were compared with values of measured free calcium (CaF) in 55 samples from normal controls and 404 samples from patients with various disorders of calcium metabolism. Three criteria were used to compare either CaC or CaT with measured CaF: 1) the correlation coefficient, 2) the average absolute deviation from measured CaF of the values of CaF predicted by the linear regression of CaF on each CaC, and 3) the number of samples in which CaC or CaT gave a different impression of normality than measured CaF. Application of the 13 published algorithms produced varied results, but none produced substantially better agreement between CaC and CaF than was found between CaT and CaF. The application of additional algorithms derived by multiple linear regression using our data base gave slightly better results than any of the published algorithms, but many values of CaC remained which were disparate from the measured value of CaF. Correction of measured total calcium by using other concurrently obtained chemistry values does not seem to adequately predict calcium status as measured by free calcium.


The arteriographic manifestations of Takayasu's arteritis in children. Eighteen cases of Takayasu's arteritis in children under 14 years of age are described, emphasising the arteriographic features and noting differences with other series. Renal artery stenosis resulting in renal hypertension was the most significant finding but widespread involvement of the aorta and its main branches with predominantly stenotic lesions was also found. Total aortography is mandatory in these cases as without it the correct diagnosis cannot be made.


Current status of semen preservation in the ram, boar and stallion. From the studies cited it was concluded that short and long term preservation of stallion semen has encountered major obstacles. Fertilizing capacity of extended or extended and cooled spermatozoa has been impaired. With the hydrogen ion extenders, the fertility was depressed either with or without glycerol when the semen was inseminated immediately after extension. With the cream-gel extender, fertility was not impaired when inseminated immediately after extension, but was impaired after storage at 5 C for 24 hr or in the presence of glycerol. The fertilizing capacity of extended frozen spermatozoa particularly from some stallions has been more adversely affected than that of others. These studies show that the pregnancy rate range was from 50 to 80% for raw semen from the same stallion used in the frozen studies. Pregnancy rate with this magnitude of difference must be carefully weighed in applying the results from a few stallions to the population. Sufficient information has been generated to suggest that the preservation of stallion spermatozoa is possible but the fertilizing capacity is impaired. Causes of this impairment must be further investigated. When this is accomplished, the number of motile spermatozoa needed per insemination and the frequency of insemination required for optimal fertilization reported in this review must then be reevaluated.


Requirements for blastocyst development in vitro. Four characteristics of culture medium that are important to embryo development and nutrition of the blastocyst have been discussed. An examination of several of the most commonly used media for embryo culture demonstrates many similarities among them. The milliosmolarities of the media range from the hypoosmotic optimums (256 milliosmols) demonstrated in several in vitro studies to the physiologic range (308 to 315 milliosmols). Media between these extremes generally allow good development. Low oxygen concentrations (5%) in the culture environment allow somewhat better development of early cleavage stages, but recent studies suggest the difference between development in 5 and 20% oxygen to be less than originally thought. The media most commonly employed for early embryo culture contain bicarbonate as the buffer, but maintenance of pH is probably not the most crucial role of the CO2-bicarbonate content of the media. Likewise, since 1965 almost all media used to culture embryos have used pyruvate as the primary energy source. This is particularly important when early stages, before blastocyst development, are cultured. The concentration used generally falls within the optimum range of 2.5 to 5.0 X 10(-4)M first reported. Although glucose is not oxidized well by the early cleavage stages, it is an important energy source for all blastocysts. Furthermore, glucose contributes more than any other carbon source, including amino acids, to protein formation. Much is yet to be learned concerning the nutrition of the blastocyst, but our knowledge has increased immensely during the last 15 years. Hopefully our progress will be at least as rapid in the coming decade.


Possible changes in striatal and limbic cholinergic systems in schizophrenia. Enzymes concerned with neurotransmitter metabolism were measured postmortem in 50 regions from the brains of 11 chronic schizophrenics, 2 patients with senile dementia, 1 depressive, and 18 controls. Enzymes studied were tyrosine hydroxylase, dopa decarboxylase, glutamic decarboxylase, choline acetyltransferase (CAT), and acetylcholinesterase. The schizophrenic group had high CAT activities in the hippocampus, caudate, putamen, and nucleus accumbens; the other patients from the same hospital did not. A compensatory response to long- or short-term drug usage is considered, but correlations are hard to establish in the group studied. An alternative hypothesis proposes that the high levels are a compensatory response to defective cholinergic receptors in the affected areas. On this hypothesis, and by analogy with chorea, dopaminergic antagonists would act in schizophrenia by helping to reestablish cholinergic-dopaminergic balance.


Observations on biphasic bronchial reactions due to inhalation of enzymes of Bacillus subtilis. The effect of disodium cromoglycate, thiazinamium and prednisone on immediate and late type allergic bronchial reactions was studied in four patients with biphasic bronchial obstruction due to inhalation of Bacillus subtilis enzymes. The immediate reaction (type 1 allergy) was not influenced by any of these drugs. Disodium cromoglycate seemed to ameliorate the late reaction, which was very much depressed by prednisone. Thiazinamium, an antihistamine drug, did not alter the immediate nor the late phase of the reaction. The data suggest that the pathogenetical mechanism involved in Bacillus subtilis allergy may differ from that initiating bronchial reactions to pollen or house dust.


Mandibular fractures. II. A follow-up study of 229 patients. During the period 1964-73, 286 patients were treated for mandibular fractures. 229 patients (80%) attended the follow-up examination. The length of observation was from 1 to 9 years. The treatment and the follow-up evaluations in the 229 patients are described. Early fracture treatment has been the objective, also in patients who have undergone severe cerebral traumas. Antibiotic prophylaxis was implemented in cases of compound fractures. Preservation of teeth and tooth buds in the line of fracture was attempted. Clinical infection occurred in 0.4% of the patients. At the follow-up examination 1 patient presented with a fracture displacement outside the condylar process. Neither malocclusion nor pseudarthrosis following fracture or after treatment were seen. Permanent sensory disturbances in the innervation area of the mental nerve following fracture occurred in 8%. Radiological examination of the teeth in the line of fracture revealed unnoticed apical bone lesions in 17% of 118 patients. In a further 23% there was a negative response to the vitality test. Follow-up control of mandibular fractures is advised at 12 to 18 months after treatment.


Relation between cell wall turnover and cell growth in Bacillus subtilis. The kinetics of cell wall turnover in Bacillus subtilis have been examined in detail. After pulse labeling of the peptidoglycan with N-acetylglucosamine, the newly formed peptidoglycan is stable for approximately three-quarters of a generation and is then degraded by a process that follows first-order kinetics. Deprivation of an auxotroph of amino acids required for protein synthesis results in a cessation of turnover. If a period of amino acid starvation occurs during the lag phase of turnover, then the initiation of turnover is delayed for a period of time equivalent to the starvation period. During amino acid starvation, new cell wall peptidoglycan is synthesized and added to preexisting cell wall. This peptidoglycan after resumption of growth is also subject to degradation (turnover). It is suggested that cell wall turnover is dependent on cell growth and elongation. Several possible control mechanisms for cell wall autolytic enzymes are discussed in light of these observations.


Regulation of accumulation and turnover of an inducible glutamate dehydrogenase in synchronous cultures of Chlorella. Earlier studies indicated that the gene of an ammonium-inducible glutamate dehydrogenase (GDH) was inducible throughout the cell cycle and was expressible shortly after replication early in the S-phase in synchronous Chlorella cells growing at a rate of 13% per h in the absence of inducer. In the present study, synchronous cells cultured at the same growth rate in the continuous presence of inducer accumulated this enzyme in a linear manner, with a positive rate change observed late instead of early in the S-phase. At a growth rate of 26% per h, the positive rate change appeared to be displaced to 1.5 h before the S-phase in the next cell cycle. With 2'-deoxyadenosine, an in vivo inhibitor of deoxyribonucleic acid (DNA) synthesis, the magnitude of the positive rate change was shown to be proportional to the relative increase in DNA in the previous cell cycle. Collectively, these data support the idea that expression of newly replicated genes of this enzyme can be delayed into the subsequent cell cycle in cells in the continuous presence of inducer. Studies with cycloheximide indicated that the inducible GDH and another GDH isozyme were stable in fully induced cells in the absence of protein synthesis. However, after ammonium was removed from the culture medium, the activity of the inducible GDH decreased rapidly in vivo, with a half-time of 5 to 10 min at 38.5 degrees C, whereas the rate of accumulation of the other GDH isozyme did not change. Addition of cycloheximide, at the time of inducer removal, prevented this loss in activity of the inducible GDH. The inability to rescue the activity of the inducible GDH, by readdition of ammonium during the deinduction period, indicates that this enzyme probably underwent irreversible inactivation and/or proteolytic degradation.


Two pools of glycogen in Saccharomyces.

Two pools of glycogen in Saccharomyces. The effect of different extraction procedures on the yields of water-soluble and water-insoluble glycogen fractions from a number of Saccharomyces strains was studied by using a specific method for glycogen determination. The similarity of the yields obtained by the different procedures showed that neither form of glycogen is an artifact, and variations in the relative amounts of glycogen in the two fractions during cell growth and in different yeast strains suggest that they represent different pools of storage material with specific roles in cell development and differentiation. A proportion of the water-insoluble glycogen fraction, solubilized by mechanical agitation, was shown to be strongly associated with a beta-glucan-like polysaccharide that may be a cell wall component.


Enzymatic methyl esterification of Escherichia coli ribosomal proteins. Enzymatic methyl ester formation in Escherichia coli ribosomal proteins was observed. Alkali lability of the methylated proteins and derivatization of the methyl groups as methyl esters of 3,5-dinitrobenzoate indicate the presence of protein methyl esters. The esterification reaction occurs predominantly on the 30S ribosomal subunit, with protein S3 as the major esterified protein. When the purified 30S subunit was used as the methyl acceptor, protein S9 was also found to be esterified. The enzyme responsible for the esterification of free carboxyl groups in proteins, protein methylase II (S-adenosyl-L-methionine:protein carboxyl methyltransferase, EC 2.1.1.24), was identified in E. coli Q13. This enzyme is extremely unstable when compared with that from mammalian origin. By molecular sieve chromatography, E. coli protein methylase II showed multiple peaks, with a major broad peak around 120,000 daltons and several minor peaks in the lower-molecular-weight region. Rechromatography of the major enzyme peak showed activities in several fractions that are much lower in molecular weight. The substrate specificity of the E. coli enzyme is similar to that of the mammalian enzyme. The Km value for S-adenosyl-L-methionine is 1.96 X 10(-6) M, and S-adenosyl-L-homocysteine was found to be a competitive inhibitor, with a Ki value of 1.75 X 10(-6) M.


Isolation and characterization of a polynucleotide phosphorylase from Bacillus amyloliquefaciens. Bacillus amyloliquefaciens BaM-2 produces large amounts of extracellular enzymes, and the synthesis of these proteins appears to be dependent upon abnormal ribonucleic acid metabolism. A polynucleotide phosphorylase (nucleoside diphosphate:polynucleotide nucleotidyl transferase) was identified, purified, and characterized from this strain. The purification scheme involved cell disruption, phase partitioning, differential (NH4)2SO4 solubilities, agarose gel filtration, and diethylaminoethyl-Sephadex chromatography. The purified enzyme demonstrated the reactions characteristic of polynucleotide phosphorylase: polymerization, phosphorolysis, and inorganic phosphate exchange with the beta-phosphate of a nucleotide diphosphate. The enzyme was apparently primer independent and required a divalent cation. The reactions for the synthesis of the homopolyribonucleotides, (A)n and (G)n, were optimized with respect to pH and divalent cation concentration. The enzyme is sensitive to inhibition by phosphate ion and heparin and is partially inhibited by rifamycin SV and synthetic polynucleotides.


The value of pulsus paradoxus in assessing the child with status asthmaticus. The presence of pulsus paradoxus (PP) in 13 episodes of status asthmaticus in 12 children, ages 13 months to 15 years, was compared sequentially to a clinical score, peak expiratory flow rate (PEFR), heart rate, arterialized capillary pH, carbon dioxide pressure (PCO2), and the ratio of inspired oxygen to oxygen pressure (FIO2)/PO2) during the first 48 hours following admission. There was a significant correlation (P less than .01) between the presence of a PP (greater than or equal to 5 mm Hg) and the clinical score (r = .79), PEFR (r = .55), and heart rate (r = .49). This was particularly striking when the PP was greater than or equal to 20 mm Hg. There was no significant correlation between the mean PP and the PCO2 or FIO2/PO2 ratio. However, a mean PCO2 exceeding 40 mm Hg was associated with a highly significant (P less than .005) difference in mean PP (22.2 mm Hg) compared to the mean PP (12.2 mm Hg) when the PCO2 was below 40 mm Hg. Although the PP technique can easily be learned by physician and nursing personnel, there are potential problems. The difficulties in children are compared to those in adults. The PP is a valuable clinical tool in assessing the severity of airway obstruction in status asthmaticus. The presence of a PP, particularly greater than 20 mm Hg, is associated with moderate to severe airway obstruction. In conjunction with the overall clinical status of the patient and frequent blood gas determinations, the PP allows for better evaluation of the patient with status asthmaticus.


Systolic hypertension and cardiac mortality of Takayasu's aortoarteritis. Long-term observations of a 35-year-old woman who died from refractory congestive heart failure due to Takayasu's aortoarteritis are reported. Severe systolic hypertension was documented in the pre-pulseless phase. After the loss of all limb pulses, but relative sparing of carotid vessels, serial ophthalmodynamometric retinal pressures suggested that central aortic pressure remained high during her terminal cardiac illness. Postmortem pathologic examination showed a narrowed, severely atherosclerotic aorta with variable occlusions of all branch vessels. The loss of capacitance and volume of the aorta appears to be the cause of systolic hypertension and refractory pump failure of the left ventricle.


Takayasu's arteritis: an unrecognized cause of pulmonary hypertension. Pulmonary arteritis and secondary pulmonary hypertension are common in patients with Takayasu's arteritis. These sequelae of arteritis have not been previously emphasized in the literature. In some patients, particularly young and middle-aged women, disproportionately severe involvement of the pulmonary arteries may be the cause of pulmonary hypertension of unknown etiology.