Commitment to community mental health aftercare services: staffing and structural implications. This study examines the views of community mental health practitioners toward aftercare services and deinstitutionalized patients. A survey of 361 practitioners in Allegheny County, Pennsylvania, finds that these views are related to practitioner experience in psychotherapeutic practice, professional affiliation, educational level, and amount of specialization in aftercare. Implications are drawn for staffing and structuring community mental health aftercare services in ways that are likely to enhance practitioner committment to aftercare.


Reduction of adrenocorticotropic hormone (ACTH) and cortisol in drug addicts treated by acupuncture and electrical stimulation (AES). Forty-two heroin addicts and 31 normal persons were examined for the effect of acupuncture and electrical stimulation (AES) on plasma ACTH, cortisol and cyclic-AMP levels. Both ACTH and cortisol levels were reduced significantly in the addicts after treatment whereas no such significant reduction was observed in the normals. Plasma cyclic-AMP level was not affected in either group. Taken together, results from the present study suggest that the mechanism of AES in the treatment of addiction may have a neuroendocrinological basis. This hypothesis is particularly attractive in view of the isolation of opiate-like peptides from the brain.


Endocrinological aspects of PTH metabolism in the kidney. Various methods for the measurement of intact PTH, PTH fragments and PTH binding to receptors were applied to clarify the role of renal receptors for the bioexpression of PTH. Glomerular receptors may contribute approximately 20% of the renal PTH catabolism. PTH also binds to tubular receptors at the luminal side (brush border membranes) as well as at the antiluminal site (basal-lateral membrane). Scatchard plot analysis of PTH binding to these receptors allows the calculation of an equilibrium dissociation constant and binding capacity, if correction is made for inactivation of bioactive PTH (measured by LAMA). Binding to receptors is--in our systems--invariably associated with degradation of the intact hormone to N-terminal and C-terminal fragments of different molecular weight. Antibodies against basal lateral membranes and against brush border membranes are able to inhibit PTH binding to tubular membranes. Sera of uremic patients with inappropriately high PTH contain a globulin which interferes with PTH binding to the receptor, suggesting that uremia may be another example for a "hormone receptor antibody disease".


A concept for the control of kidney production of erythropoietin involving prostaglandins and cyclic nucleotides. Our hypothesis is that PGs released within the kidney play a role in the modulation of kidney production of Ep. PGs release probably at medullary and/or cortical sites following erythropoietic stimuli such as hypoxic hypoxia, anemic hypoxia, and ischemic hypoxia induced by renal artery constriction increase kidney production of Ep. PGs which are released probably activate a renal cortical adenylate cyclase thereby enhancing the production of intracellular cAMP. This initiates the cascade of events resulting in the production and/or secretion of Ep by the kidney. The endoperoxide analogs and PGE2 have been found to produce a dose-related and Ep-dependent increase in radioiron incorporation into newly formed red blood cells of exhypoxic polycythemic mice. Indomethacin, a potent PG cyclo-oxygenase inhibitior, attenuates Ep production and the appearance of PGE in the renal venous effluent of animals exposed to hypoxic hypoxia and renal artery constriction. Arachidonic acid (C20:4) and PGE2 infusion into the posthypoxic isolated perfused dog kidney produced a significant elevation in Ep titers in the perfusate. The increase in Ep production caused by arachidonate is blocked by indomethacin. It has been previously reported that PGs of the E series stimulate cAMP formation in several tissues. We have found that not only are renal cortical cAMP levels significantly elevated in rats following exposure to hypobaric hypoxia but that dibutyryl cAMP administration produces an increase in hematocrit and circulating red cell mass in normal mice. Our data thus far strongly support the hypothesis that the renal PGs and the cyclic nucleotides are intimately involved in the pharmacologic and/or pathophysiologic control of Ep production. Further work is necessary to determine whether the PGs and cyclic nucleotides are involved in the day-to-day control of Ep production by the mammalian kidney.


Diseases transmitted from pets to man: an evolving concern for veterinarians. Pets are not a major source of human infections but they can transmit certain diseases to man. This transmission usually is complex, requiring close contact with pets or their excretions and frequently involves a breach of sound hygienic practice. In some instances, pathogens of animal origin are acquired inadvertently because  infectivity can persist after evidence of gross contamination has gone. Veterinarians participate in controlling zoonotic diseases by encouraging rabies vaccination and hygienic treatment of pet feces and urine, by supporting community efforts toward responsible pet ownership and by advising on precautions for handling sick animals. It is recommended that veterinarians discourage the keeping of wild or exotic animals as pets and excess fondling of pets (particularly by children and pregnant women). Clients and kennel workers should be advised to use caution with animals that have aborted.


Cerebral granular cell tumor (myoblastoma) in a dog: case report and literature review. An infiltrative granular cell tumor (myoblastoma) occurred in the superficial parieto-occipital cortex of an ataxic 12-year-old dog. The neoplasm was characterized by cells with numerous small PAS-positive, diastase-resistant cytoplasmic granules. The neoplasm also had numerous cells with large globular PAS-positive bodies (angulate bodies). Ultrastructurally, the neoplastic cells were characterized by numerous homogenous electron-dense, sharply bounded granules and large pleomorphic, indistinctly bounded granules. The angulate bodies were composed of 8- to 10-nm fibrils, a few of which formed parallel arrays or tubule-like structures. This represents the first report of a granular cell tumor in the central nervous system of the dog.


Species differences in the hepatic response to mirex: ultrastructural and histochemical studies. Mirex was fed at levels of 1, 5, 15, and 30 ppm to mice, 5 and 30 ppm to rats, while monkeys received the chemical by stomach tube at levels of 0.25 and 1 mg/kg (equal to 5 and 20 ppm). Mice were killed and their livers obtained at 2, 4, 6, 9, 10, 15 and 18 months, whereas rats were killed and surgical biopsies were taken at 16, 19, 26, and 36 months. Cytochemical techniques were employed to detect activities of lysosomal beta-glycerol phosphatase (ACpase) and glucose 6-phosphatase (G-6-pase). ACpase and G-6-pase remained unchanged and comparable to controls in livers of mice receiving 1 ppm. G-6-pase decreased in centrilobular areas while ACpase increased with time in the higher groups. At 12 months, liver cells that lost their G-6-pase activity surrounded by Kupffer cells that contained strong ACpase. In contrast, rat livers had no increase in ACpase and little loss in G-6-pase. Surprisingly, and in spite of the high levels of mirex ingested, monkey livers showed no loss of G-6-pase or activation of ACpase. Ultrastructurally, the underlying feature in all livers was proliferation of smooth endoplasmic reticulum (SER) displaying species variation. Thus, in mice, intense proliferation of SER that was both time- and dose-dependent was localized in specific regions of the cytosol. Hepatic cells, damaged and necrotic in mice fed 5, 15, and 30 ppm, were phagocytosed by activated Kupffer cells. SER proliferation in rat and monkey liver cells was less conspicuous than in mice. Except for this change, rat and monkey liver cells were normal. There studies emphasize species and enzyme variations in response to mirex. An interesting aspect observed was the lack of lysosomal catabolism during liver enlargement.


[Evolution over time of the electroencephalographic signal. Application in pharmacology]. The dynamics of transition from wakefulness to drowsiness can be modified by psycho-active compounds. These two states of vigilance have different electroencephalographic characteristics, which can be also modified by the administration of substances. This paper describes a method intended to discriminate between these two aspects of drug action, using a mini-computer. Investigation of sleep-wakefulness cycles with continuous EEG and EMG recording necessitates an efficient method of data reduction. On-line computation of Hjorth parameters with a micro-computer, and further processing witha mini-computer has been used. A pharmacological approach to the study of sleep-wakefulness cycles is proposed; this avoids a strict distinction of sleep phases, and concentrates on the coninuum of vigilance states. Differents ways of showing drug-induced modifications on that continuum are presented with examples.


Gonadotropin induced stimulation and desensitization of cyclic 3', 5' -adenosine monophosphate dependent protein kinase(s) in the rat ovary. Incubation of rat ovarian slices with choriogonadotropin resulted in stimulation of cyclic AMP dependent protein kinase activity in a time and dose dependent manner. However, prior exposure of animals to choriogonadotropin in vivo resulted in a decreased sensitivity to subsequent in vitro stimulation by the same hormone. This decreased sensitivity was due to a lesion in the hormone receptor adenylate cyclase system rather than a change in the property of protein kinase with respect to its stimulation by cyclic AMP. Unchanged levels of cyclic AMP formation in these ovaries were evidenced by the lack of inhibition of [3H] cyclic AMP binding. In the untreated controls, however, choriogonadotropin in vitro resulted in inhibition [3H] cyclic AMP binding activity suggesting endogenous occupation parallel to protein kinase activation. These results are indicative of the linear coupling of hormone receptor-adenylate cyclase-protein kinase system and point out the in vivo role of protein kinase activation in choriogonadotropin mediated regulation of ovarian function.


Corticosteroid response of rabbits and rats to exogenous ACTH. The dynamics of the alterations of the cortisol: corticosterone ratio in rabbits during porcine ACTH administration were studied on an almost daily basis and as a function of time after injection. In rabbits the cortisol: corticosterone ratio increased strikingly but variably during the treatment period. Rats responded to similar treatment only with increased corticosterone release. The differences are attributed to the presence of 1-39 ACTH as well as intermediate ACTH in the rabbit pituitary but not in the rat pituitary.


Characterization of specific prostaglandin E receptor in isolated rat Leydig cells. The binding of 3H-prostaglandin E1 (PGE1) to the isolated Leydig cells from rat testes was studied under different experimental conditions. The Scatchard plot analysis showed two populations of specific binding sites: one of a high affinity and low capacity (Ka1 = 2.2 x 10(8) mol-1 l) and the other with less affinity and high capacity (Ka2 = 8.7 x 10(7) mol-1 l). Maximum specific binding occurred at pH 7.4 and 22 degrees C. Unlabelled PGE1 and PGE2 inhibited the binding in a dose-dependent manner, while about 10 times higher amount of PGA1 was necessary to show a similar effect. Moreover, the treatment of Leydig cells with disulfide modifying agents (dithiotreitol and mercaptoethanol) decreased 3H-PGE1 binding. Finally, the production of cAMP was 4.5 times increased (P less than 0.001) after 20 min incubation of Leydig cells with PGE1 and aminophylline, while that of testosterone was 2.5 times increased (P less than 0.001) after 3 h incubation with PGE1.


Activity of adenylate cyclase and phospodiesterase in the pituitary and thyroid during postnatal period in rats. Adenylate cyclase (AC) and phosphodiesterase (PD) activities in the pituitary and thyroid have been determined during postnatal period in rats. In both tissues the activity of both enzymes was found early after birth. In the pituitary gland AC activity increased significantly from the second to the seventh day, while it remained unchanged on the 16 th day and again increased on the 23rd day when the activity reached the value of adult rats. The activity of PD in the pituitary increased significantly from the second to 23rd postnatal day and in the adult rats the activity was similar as in two days old rats. In the thyroid gland AC activity remained unchanged during postnatal period of rats with the exception between the 16th and 23rd day when a small but significant difference was found. The activity of PD in thyroid was high on the first day after delivery and then decreased significantly on the seventh day. It increased again significantly on 23rd day when it was similar to the adult rats. However, between 23rd and 60th days the activity significantly dropped and significantly increased from 60th day to the adult age. These results suggest that in the rat the mechanism of adenylate cyclase system involved in the pituitary and thyroid functions could play a role in the regulation of the function of these tissues during postnatal development.


Studies on adrenaline-induced lipolysis in adrenalectomized rats. In a short-term study, adrenaline-induced lipolysis was less in adrenalectomized rats than in controls, though the cyclic AMP accumulation was not different. In adrenalectomized rats treated with corticosterone, lipase activity was as low as in untreated adrenalectomized rats, although adrenaline-induced lipolysis was not reduced. In a long-term study, no reduction in adrenaline-induced lipolysis or cyclic AMP accumulation was observed in adrenalectomized rats. The mechanism of the effect of adrenalectomy on adrenaline-induced lipolysis is discussed on the basis of these results.


Changes in the cyclic nucleotides of rat thyroid, pituitary and plasma caused by methylthiouracil treatment. Changes in the content of cyclic nucleotides (cAMP and cGMP) and related enzyme activities were observed in the rat thyroid, pituitary and plasma during the prolonged increase of endogenous TSH produced by treatment with methylthiouracil (MTU). Experiments were performed after 4 weeks treatment with MTU. The wet weight and cAMP content per wet weight of the thyroid increased 3 and 1.4 times respectively, but cGMP showed a slight decrease. Pituitary weight increased 1.3 times, but cAMP and cGMP content did not change. The cAMP level in plasma also increased about 1.3 times, but cGMP did not increase. The cAMP-phosphodiesterase activity in the thyroid, pituitary and plasma was increased 1.9, 1.4 and 1.3 times respectively after MTU treatment, while cGMP-phosphodiesterase showed no significant change. ATPase activity in the thyroid and pituitary was also increased more than 1.5 times after MTU treatment, while 5'-nucleotidase activitity decreased remarkably. These data indicate that the metabolism of the cyclic nucleotide system in the thyroid is stimulated by TSH.


Enzymatic oxidation of mercaptoethanol to isethinic acid and isethionic acid. The enzymatic oxidation of mercaptoethanol by purified cysteamine oxygenase has been studied. Products were identified by chromatography as isethinic acid (2-hydroxyethan-sulfinic acid) and isethionic acid. Other features of the reaction, as cofactor requirement, the influence of the enzyme concentration on the stage of oxidation, the biological significance of this reaction are also discussed.


beta-Glucuronidase activity in trained red and white skeletal muscle of mice. We studied the effects of prolonged running exercise (5 days a week, 1.5 h per day at a speed of 17.6 m/min) on the activity of some acid hydrolases (beta-glucuronidase, beta-N-acetylglucosaminidase, acid phosphatase and cathepsin D) and three enzymes of energy metabolism (cytochrome c oxidase, lactate dehydrogenase and creatine kinase) in the distal and in the proximal, the predominantly white and red parts, respectively, of the vastus lateralis-muscle from mice. The acid hydrolase activity levels were 1.24--1.69 higher in untrained red muscle compared to untrained white muscle. The light training applied increased the activity of beta-glucuronidase in both red and white muscle. No other significant training effects were observed in the enzyme activities measured.


Immunological studies on cytochrome c oxidase: arrangements of protein subunits in the solubilized and membrane-bound enzyme. Seven protein subunits of cytochrome c oxidase from bovine heart were isolated by gel filtration in the presence of sodium dodecyl sulphate (subunits I, II and III) and guanidine hydrochloride (subunits V, VI and VII), and ion-exchange chromatography in 6 M urea (subunit IV) after the enzyme had been dissociated in 6 M guanidine hydrochloride. When analysed by highly cross-linked sodium dodecyl sulphate/polyacrylamide gel electrophoresis in the presence of urea, the apparent molecular weights were = I, 36700; II, 24300; III, 20400; IV, 17300; V, 12300; VI, 8700: and VII, 5100. Monospecific rabbit antisera were obtained against subunits I, IV, V, VI and VII and a mixture of subunits II and III. These subunit-specific antisera with the exception of anti-I serum all cross-reacted with the detergent-solubilized native oxidase. Enzymatic studies on purified oxidase indicated that immunoglobulins against subunits II + III, IV, V, VI and VII respectively caused 25, 65, 20, 30 and 25% inhibition while anti-I immunoglobulin did not inhibit the activity. The subunit-specific antisera were used to examine the arrangements of the subunits in the membrane. Enzymatic studies using bovine heart mitochondria and rat liver mitochondrial digitonin particles showed that anti-(II + III) serum, anti-V serum and anti-VII serum all inhibited the oxidase activity while the other antisera did not. On the other hand, results of using 125I-labelled immunoglobulins showed that anti-IV, anti-V and anti-VII sera were bound to the surface of inverted vesicles (matrix side) while all other antisera were not. These results indicate that cytochrome oxidase subunits II and III are situated on the outer surface, and subunit IV is exclusively on the matrix surface while subunits V and VII are exposed on both surfaces of the mitochondrial membrane. Subunits I and VI are buried within the membrane, not exposed on either side.


Partial purification and characterization of rat-liver messenger RNA coding for phosphoenolpyruvate carboxykinase (GTP). The mRNA coding for the gluconeogenic enzyme phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) was partially purified from the liver of cyclic-AMP-treated rats by a procedure involving multiple oligo(dT)-cellulose chromatographies and sucrose gradient fractionations. The purification was monitored by translational assay using a wheat germ extract. Relative to RNA bound once to oligo(dT)-cellulose, the final material was enriched 20-fold in template activity for phosphoenolpyruvate carboxykinase synthesis. With this RNA preparation, cell-free enzyme synthesis amounted to 5% of total mRNA-directed protein synthesis. The apparent sedimentation coefficient of phosphoenolpyruvate carboxykinase mRNA in sucrose gradients was between 20 and 22 S, corresponding to an average molecular weight of 0.93 X 10(6). By formamide/polyacrylamide gel electrophoresis the molecular weight of the enzyme mRNA was estimated at between 0.91 X 10(6) and 1.12 X 10(6). From these estimates, it was concluded that considerable non-coding sequence(s) are present in the mRNA. Approximately 20% of the enzyme mRNA in rat liver failed to bind to oligo(dT)-cellulose, presumably because of the absence of a poly(A) segment. The translation of phosphoenolpyruvate carboxykinase mRNA by the wheat germ extract was inhibited in the presence of 7-methylguanosine 5'-phosphate. The enzyme mRNA appears therefore to have a 'cap' at the 5' end.


Modulation of adenylate cyclase/cyclic AMP response by thyrotropin and prostaglandin E2 in cultured thyroid cells. 1. Negative regulation. Isolated porcine thyroid cells, cultured in the presence of thyrotropin (greater than or equal to 0.25 mU/ml) or prostaglandin E2 (greater than or equal to 0.1 micron), showed decreased adenosine 3':5'-monophosphate (cyclic AMP) response to further thyrotropin or prostaglandin E2 stimulation, respectively. Kinetics of the refractory process to thyrotropin and prostaglandin E2 are different: (a) maximal refractoriness to prostaglandin E2 was attained after 2--6 h exposure to prostaglandin E2 while refractoriness to thyrotropin was maximal only after 12--24 h; (b) the degree of refractoriness to prostaglandin E2 was much greater than that to thyrotropin. Refractoriness to thyrotropin or prostaglandin E2 is characterized: by specificity for each thyroid stimulator; by dependence upon the dose of thyrotropin or prostaglandin E2 in culture, e.g. induction of high degree of refractoriness with 0.5 mU/ml thyrotropin (or 1 micron prostaglandin E2), which elicits only a small cyclic AMP increase; by time requirement for induction; by partial effect; by changes of maximum activation of cyclic AMP response; by reversibility. This refractoriness of the cyclic AMP response was not induced by dibutyryl adenosine 3':5'-monophosphate. It was not attributed to increased cyclic AMP-phosphodiesterase activity, but to alterations in the receptor-adenylate cyclase system. Prevention of refractoriness to thyrotropin or prostaglandin E2 by incubation of cells in the presence of actinomycin D, puromycin and cycloheximide suggests that new RNA and protein syntheses are required for the development of the refractory state.


Modulation of adenylate cyclase/cyclic AMP response by thyrotropin and prostaglandin E2 in cultured thyroid cells. 2. Positive regulation. Two different independent processes are operating in cultured thyroid cells to regulate adenylate cyclase/cyclic AMP responsiveness to thyroid stimulators (thyrotropin and prostaglandin E2): firstly, refractoriness or negative regulation [preceding paper], which is specific for each thyroid stimulator, is not mediated by cyclic AMP and is not accompanied by alteration of adenylate cyclase activity; secondly, positive regulation which is characterized by an augmentation of the cyclic AMP response stimulated by thyrotropin and prostaglandin E2. This process is not specific for each thyroid stimulator and is a state of increased susceptibility of cyclic AMP synthesis to stimulation, accompanied by increased activity of the catalytic subunit of adenylate cyclase. Positive regulation is apparently mediated by increased intracellular cyclic AMP levels. It is a time-dependent and dose-dependent process. Very low concentrations (5-50 micronU/ml) of thyrotropin augmented cyclic AMP synthesis stimulated by thyrotropin and prostaglandin E2 whereas higher concentrations (above 0.1 mU/ml) augmented prostaglandin E2 stimulation but induced refractoriness to thyrotropin. Prostaglandin E2 (0.1 to 10 micronM) augmented thyrotropin stimulation and dibutyryl adenosine 3':5'-monophosphate (0.3 to 2 mM) augmented thyrotropin and prostaglandin E2 stimulation. Positive regulation is a slow process which develops within days and increases up to day 5 in culture. Experiments using inhibitors suggested that protein synthesis is required for the full expression of the increase in adenylate cyclase activity induced by the studied thyroid stimulators.


Carbon-13 nuclear-magnetic-resonance spectra of adenine cyclonucleosides and their phosphates. Effects of neighboring groups for elucidation of fine structure of nucleosides and nucleotides. Carbon-13 nuclear magnetic resonance spectra of adenine cyclonucleosides, which have a fixed glycosidic conformation in an anti range, and their isopropylidene and phosphate esters are reported; those of 9-beta-D-arabinofuranosyladenine and its 5'-phosphate are also presented. The chemical shifts of the base carbons are affected not only by the bridging atom but also by the position of the bridged sugar carbon which determine the planarity of the third ring formed by cyclization between the base and the sugar. The effects of glycosidic conformation on the sugar-carbon chemical shifts are discussed by comparison of the data for 8:5'-cycloadenosines with the data for adenosine and its 8-substituted derivatives. The effects of a 2'-oxygen on sugar-carbon chemical shifts have been examined by comparing the data for 2'-deoxyadenosine, arabinosyladenine and 8:2'-anhydro-8-oxy-9-beta-D-arabinofuranosyladenine. The effects of phosphomonoester groups on base and sugar carbon resonances have been examined and it is noted that these groups cause downfield shifts for C-8 of all cyclonucleotides. Data for the 3':5'-cyclic monophosphate derivative of 8:2'-anhydro-8-thio-9-beta-D-arabinofuranosyladenine suggest that the previous assignments of C-4' and C-3' for nucleoside 3':5'-cyclic monophosphates must be reversed. According to the reversed assignments, it seems that C-3' and C-5' show moderate downfield shifts and C-4' shows a marked upfield shift.


Isolation of precursors of cytochrome oxidase from Neurospora crassa: application of subunit-specific antibodies and protein A from Staphylococcus aureus. A novel immunological procedure has been applied for the isolation of precursors of cytochrome oxidase. It involves antibodies to individual subunits of the oxidase and protein A from Staphylococcus aureus linked to a Sepharose support. Unassembled (free) 'subunits' as well as a labile complex containing five polypeptide components of the oxidase were isolated from mitochondrial extracts by this technique. The procedure is superior to the previously used double-immuno-precipitation method, because of its quantitative nature, its sensitivity, rapidity and versatility. Thus, sequential titrations of precursor proteins by addition of various subunit-specific immuno-globulins to an individual extract become feasible. Furthermore, the technique is suitable for the isolation of precursors on a large scale. To avoid contamination of the polypeptide preparations with immunoglobulin, the antibodies were covalently coupled to protein A, which had been previously linked to Sepharose. A radioactive preparation of unassembled subunit 1 of cytochrome oxidase was isolated by such a modified support and its cyanogen-bromide-cleavage products were compared to those of subunit 1 obtained from the assembled enzyme.


Comment on the use of linear free-energy slopes as criteria for distinguishing hydride-ion from hydrogen-atom transfer in reactions of pyridine-nucleotide coenzymes. For reductions by dihydropyridines, the slopes of plots of log k vs the standard reduction potential of the dihydropyridine are not direct indicators of whether the rate-determining step is hydride-ion or hydrogen-atom transfer. When there is no substituent effect on the reverse rate, those slopes are independent of mechanism and equal to about (30mV)-1.


The reaction of pyruvate with saccharopine dehydrogenase. The preceding paper in this journal has reported that pyruvate could be substituted for 2-oxo-glutarate as a substrate of saccharopine dehydrogenase [epsilon-N-(L-glutaryl-2)-L-lysine:NAD oxidoreductase (L-lysine-forming) in the direction of reductive condensation. In the present communication, the kinetic mechanism of saccharopine dehydrogenase reaction with NADH, L-lysine and pyruvate as reactants is reported. The results of initial velocity study, inhibition studies with lysine analogs and a reaction product, NAD+, are consistent with an ordered mechanism with the coenzyme binding first and pyruvate last. The reaction mechanism is at variance with that of the normal reaction in which 2-oxoglutarate is the substrate, in that the order of addition of the amino and  oxo acid substrates is reversed. This fact suggests that there exists a small degree of randomness in the binding of amino and oxo acid substrates. From a product inhibition study, NAD+ was shown to be the last reactant released. Saccharopine [epsilon-N-(L-glutaryl-2)-L-lysine] was found to act as a potent dead-end inhibitor of the condensation reactions (of lysine and 2-oxoglutarate, and of lysine and pyruvate) by forming an abortive E. NADH. saccharopine complex.


Purification and properties of poly(ADP-ribose) polymerase from pig-thymus nuclei. The nuclear enzyme poly(ADP-ribose) polymerase has been purified about 9200-fold from pig thymus nuclei with a 46% yield. An aqueous organic solvent system was used for the isolation of the polymerase from nuclei and for its purification by chromatography at sub-zero temperatures. Electrophoretic analysis under both denaturing and non-denaturing conditions revealed a single protein band suggesting that the preparation was homogeneous and that the enzyme is composed of one polypeptide chain. The molecular weight estimated from sodium dodecyl sulphate-/polyacrylamide gel electrophoresis was 63 500 and from gel filtration through columns of Sephadex G-100, 58 000. The enzyme preparation was free from poly(ADP-ribose)-degrading enzymes and from DNA. The purified polymerase showed an absolute requirement for both DNA and histones. The maximal specific activity of the homogeneous preparation measured by the standardized assay, was 20.7 mu mol NAD+ incorporated x min-1 x mg-1 of protein at 37 degree C. Amino-terminal group analysis with dansyl chloride did not reveal a terminal amino acid suggesting that the amino-terminal group may be blocked. In the presence of histones, the Km for NAD+ was 23 micrometer.


Mapping the ATP-binding site in the catalytic subunit of adenosine-3':5'-monophosphate-dependent protein kinase. Spatial relationship with the ATP site of the undissociated enzyme. A set of 24 ATP analogs modified at various positions of the ATP molecule was used for mapping the ATP-binding site in the free catalytic subunit (C) of cAMP-dependent protein kinase (type I). Ki values for these analogs (of which 23 were shown to be competitive with ATP) were measured and compared with Ki values previously obtained for the same set of analogs upon binding to the undissociated form of the enzyme (R2C2). It was found that modifications at the adenine part of ATP bring about a considerable reduction in affinity between C and the resulting analog. The other parts of the ATP molecule play a less important, though definite, role in the binding of this nucleotide to C. By measuring the effect of each given modification in ATP on its binding to C, and comparing the effect of this modification on the binding of the same analog to R2C2, it was possible to obtain 'specificity profiles' for both forms of the kinase. Using such profiles it is shown that the adenine-binding subsite in C may well coincide with the adenine-binding subsite in R2C2. Two plausible models describing the spatial relationship between the ATP sites in C and R2C2 are proposed.


Actin amino-acid sequences. Comparison of actins from calf thymus, bovine brain, and SV40-transformed mouse 3T3 cells with rabbit skeletal muscle actin. Actin was purified from calf thymus, bovine brain and SV40-transformed mouse 3T3 cells grown in tissue culture. Isoelectric focusing analysis showed the presence of the two actin polypeptides beta and gamma typical for non-muscle actins in all three actins. Tryptic and thermolytic peptides accounting for the complete amino-acid sequence of the cytoplasmic actins were separated and isolated by preparative fingerprint techniques. All peptides were characterized by amino-acid analysis and compared with the corresponding peptides from rabbit skeletal muscle actin. Peptides which differed in amino-acid composition from the corresponding skeletal muscle actin peptides were subjected to sequence analysis in order to localize the amino-acid replacement. The results obtained show that all three mammalian cytoplasmic actins studied contain the same amino-acid exchanges indicating that mammalian cytoplasmic actins are very similar if not identical in amino-acid sequence. The presence of two different isoelectric species beta and gamma in cytoplasmic actins from higher vertebrates is acccounted for by the isolation of two very similar but not identical amino-terminal peptides in all three actin preparations. The nature of the amino-acid replacements in these two peptides not only accounts for the different isoelectric forms but also shows that beta and gamma cytoplasmic actins are the products of two different structural genes expressed in the same cell. The total number of amino-acid replacements so far detected in the comparison of these cytoplasmic actins and skeletal muscle actin is 25 for the beta chain and 24 for the gamma chain. With the exception of the amino-terminal three or four residues, which are responsible for the isoelectric differences, the replacements do not involve charged amino acids. The exchanges are not randomly distributed. No replacements were detected in regions 18--75 and 299--356 while the regions between residues 2--17 and 259--298 show a high number of replacements. In addition documentation for a few minor revisions of the amino acid sequence of rabbit skeletal muscle actin is provided.


Restriction and modification in Bacillus subtilis. Localization of the methylated nucleotide in the BsuRI recognition sequence. Calf thymus DNA was methylated in vitro with cell extracts of Bacillus subtilis OG3R (r+m+) and S-adenosyl[Me-3H]methionine. After depurination of the [3H]methylated DNA, the analysis of the pyrimidine dinucleotides revealed the following positions of the methylated nucleosides (indicated by an asterisk) within the BsuRI recognition sequence: 5' dG--dG--dC--dC dC--dC--dG--dG 5'.


Mechanism of activation of protein kinase I from rabbit skeletal muscle. The equilibrium parameters of ligand interaction and protein dissociation. Protein kinase I from rabbit skeletal muscle binds adenosine 3':5'-monophosphate (cAMP) and ATP with high affinity; cAMP promotes and ATP retards dissociation (activation) of the tetrameric enzyme. The interrelationship of ligand interaction with protein dissociation has been probed by quantitative ligand binding, using the filter assay technique, and by computer simulation of binding curves in terms of Scatchard plots. A comparison of the experimental and computed binding data strongly confirms the supposition that the dimeric regulatory subunit (R2) binds cAMP cooperatively. Mainly from ATP binding it is further concluded that the interaction of the two catalytic subunits with R2 is also strongly cooperative, whereas the binding of ATP to the holoenzyme is non-cooperative. The underlying random model, possessing low-affinity and high-affinity sites not only for ATP but also for cAMP, allows the determination of a minimal set of equilibrium parameters required for description and also an estimation of their magnitude. It further provides a basis for an explanation of the different behaviour of protein kinases I and II reported in the literature.


Lymphocyte subpopulations in blood and cerebrospinal fluid from patients with acute Guillain-Barré syndrome. The numbers of T and B lymphocytes were determined in blood and cerebrospinal fluid (CSF) from 25 patients with acute Guillain-Barré syndrome and from 30 controls. The relative and absolute number of T lymphocytes in blood was decreased in the patient group (mean 48 +/- 2% and 947 +/- 71/mm3 compared to 66 +/- 2% and 1,381 +/- 93/mm3 in controls). Correspondingly, the number of B lymphocytes was increased (mean 37 +/- 2% and 758 +/- 75/mm3 compared to 25 +/- 2% and 580 +/- 45/mm3 in controls). In the CSF the number of T lymphocytes was increased (85.0 +/- 2.2% compared to 76.9 +/- 2.4% in controls).


Latent viruses and Bell's palsy in children. 4 children with unilateral facial palsy--Bell's palsy (BP)--had serological evidence of primary infection with Epstein-Barr virus (EBV). Concomitant infection with cytomegalovirus (CMV) was demonstrated in 1 child while in all 4 children a rise in antibody titers to an additional one or two viruses was demonstrated. The viruses involved were herpes simplex, CMV and adenovirus which are latent in humans. Immunosuppression induced by EBV and steroid treatment may cause reactivation of these viruses resulting in rise in antibody titers. The need for prolonged serological follow-up for possible reactivation of latent viruses in BP is emphasized.


Night sleep organization in patients with severe hepatic failure. Its modifications after L-dopa treatment. 20 subjects with severe hepatic failure have been studied by means of all night polygraphic sleep recordings. Base night recordings have been made, as well as registrations after treatment with drugs that are known to improve the clinical conditions of these patients, especially concerning the psychic performances. Base night recordings show sleep disruption, with a large amount of wakefulness at the beginning of the registrations and during the night itself; sleep stages are strongly reduced, especially REM sleep. After L-dopa treatment, these negative patterns are furthermore enhanced and the percentage incidence of the REM stage is very low; nevertheless, this behaviour goes side by side with a light improvement in psychological performances. On the contrary, therapy with lactulose seems to improve the neuropsychiatric state and the sleep organization; as a matter of fact, wakefulness is lower in the night records after lactulose treatment, whilst the percentage incidence of sleep stages tends back toward normality; REM sleep has clearly risen.


Collagenase isolation and 45Ca efflux studies of human islets of Langerhans. 22 human pancreases were removed soon after circulatory arrest from donors aged 15--63 years. Part of the pancreas was used for isolation of islets by the collagenase technique. The mean yield +/- SEM, expressed as the number of islets isolated from 4 g pancreas was 79 +/- 15. The yield varied considerably even when the pancreas was removed immediately after circulatory arrest and the histology was normal, and there was no correlation with the age of the donors. The islet content of insulin (ng/microgram dry islet) was 8.5 +/- 1.2 (mean +/- SEM). Isolated islets were loaded with 45Ca in the presence of 20 mM glucose and placed in a perfusion apparatus for further studies of the 45Ca washout. The decrease of washout of radioactivity in Ca2+-deficient medium offers support for the existence of a Ca2+-Ca2+ exchange process. When added in a concentration of 20 mM, glucose tended to stimulate 45Ca efflux in perfusion medium of ordinary ionic composition but inhibited this process when the medium was deficient in Ca2+. Exposure to the calcium ionophore X-537A resulted in immediate stimulation of of 45Ca efflux from human islets as previously observed for islets from rats and mice. This suggests that Ca2+ has a direct regulatory role for insulin release also in humans.


The use of a test for the differential diagnosis of hypercalciuria. 28 renal stone formers (18 men and 10 women) with idiopathic hypercalciuria (IH) and 27 controls have been subjected to a test proposed for the diagnosis of absorptive, resorptive and renal hypercalciurias. Fasting serum calcium concentration, urinary calcium and cyclic AMP excretion were measured after overnight fasting and an oral load of calcium. Absorptive hypercalciuria was demonstrated in 14 patients. High fasting urinary calcium first suggested resorptive or renal hypercalciurias in 5 other patients, but since fasting urinary calcium was normalized following cellulose phosphate therapy, absorptive hypercalciuria was more likely. Renal hypercalciuria was a possibility in 1 single case. Both fasting and post-load urinary calcium were normal in 7 men and 1 woman. The test did not appear as useful as expected since it was of no diagnostic value in about 30% of the cases and erroneously suggested resorptive or renal hypercalciuria in about 15% of the cases. On the other hand it indicated that absorptive IH is common and renal IH exceptional.


Heterogeneity of cerebral beta-adrenoceptor binding sites in various vertebrate species. [3H]-Dihydroalprenolol ([3H]-DHA) binds to cerebral membranes of the frog, chick, rat, mouse, rabbit and human with a dissociation equilibrium constant (KD) of about 1 nM and displays binding characteristics indicative of an interaction with beta-adrenoceptors. However, the maximum number of specific binding sites labelled by this beta-adrenoceptor ligand varies substantially between the species with the chick and mouse having the highest, and the frog the lowest density. The structure--activity relationships of adrenergic agents to inhibit specific [3H]-DHA binding suggests that whereas the membrane sites from all the species had similar affinities for non-selective beta-adrenergic agents, several drugs that have been reported to show selectivity for beta1-adrenoceptors demonstrated considerably higher affinities for mammalian rather than avian or amphibian membrane sites. By this pharmacological criteria it is likely that all the beta-adrenoceptor binding sites in frog and chick cerebral tissue have properties resembling beta2-receptors. However, in mammalian cerebral cortex, evidence is presented that beta1- and beta2-adrenoceptors coexist in a ratio of 70%/30% respectively.


Effect of pretreatment with bronchodilator drugs on in vitro responsiveness of guinea pig lung adenylate cyclase. Pretreatment of guinea pigs with 5 microgram/kg isoprenaline or 10 microgram/kg salbutamol s.c. thrice daily for 7 days reduced the responsiveness of lung slice and tracheal ring adenylate cyclase to isoprenaline, but not to prostaglandin E1. Pretreatment of guinea pigs with isoprenaline also reduced the sensitivity of tracheal smooth muscle strip adenylate cyclase to isoprenaline. Cross-tolerance developed to noradrenaline in lung slices obtained from guinea pigs pretreated with isoprenaline. Propranolol blocked the response of lung slice adenylate cyclase of control and isoprenaline-pretreated animals to approximately the same degree. The presence of phentolamine in the incubation medium did not affect the reduced sensitivity to isoprenaline. Possible mechanisms of development of tolerance to sympathomimetic bronchodilator drugs are discussed.


Presynaptic inhibitory effects of catecholamines on cholinergic transmission in the smooth muscle of the chick stomach. In the vagus nerve--smooth muscle preparation isolated from the chick proventriculus, adrenaline, clonidine (10(-8) - 2.5 x 10(-7) M), noradrenaline (10(-7) - 2.5 x 10(-6) M) and dopamine (10(-5) - 10(-4) M) inhibited the contraction induced by low frequency (0.5 Hz) stimulation of the vagus nerve, but they did not inhibit the contraction elicited by acetylcholine (5 x 10(-8) - 5 x 10(-7) M). The concentration producing 50% inhibition was 10(-7) M for adrenaline and clonidine, 10(-6) M for noradrenaline, and 5 x 10(-5) M for dopamine. Isoproterenol (5 x 10(-8) - 5 x 10(-7) M) inhibited the responses induced by both stimulation of the vagus nerve and acetylcholine. The inhibitory effects of the catecholamine and clonidine were blocked by phentolamine (2.7 x 10(-6) M) but not by 5-(3-tert-Butylamino-2-hydroxy)-propoxy-3, 4-dihydrocarbostyril hydrochloride (OPC 1085) which blocked the effect of isoproterenol. It is suggested that presynaptic alpha-receptors are present in the myenteric plexus of the chick proventriculus, and that the catecholamines and clonidine exert their inhibitory effects on cholinergic transmission via these receptors.


Effects of diethyldithiocarbamate and fusaric acid upon memory storage processes in rats. Diethyldithiocarbamate (DDC) and fusaric acid (FA), administered immediately posttraining, disrupted retention performance of rats trained on a one-trial inhibitory avoidance task. Posttraining DDC disrupted retention performance if the treatment was delayed up to at least 24 h (but not 72 h) after training. A single DDC injection also disrupted retention performance if the treatment was administered up to 10 days prior to training. The effects of DDC and FA upon several neurobiological measures were also investigated. Amnestic doses of DDC influence brain and adrenal catecholamine levels, brain electrographic activity and hippocampal Timm's staining. On the other hand, amnestic doses of FA only affected catecholamine levels. The amnestic effects of DDC were dissociated from brain seizure activity produced by DDC. The effects of DDC and FA upon retention may have been related to the drugs' effects upon catecholamine systems, but the bleaching by DDC of Timm's staining could reflect additional processes involved in DDC's effects upon retention. As opposed to the more widespread effects of DDC, FA's relative lack of effect upon other neurobiological measures more strongly supports a catecholaminergic interpretation for its effect upon retention.


Development of sympathetic nerve--muscle transmission in chick. The development of sympathetic nerve--muscle transmission was investigated pharmacologically by using the nerve-expansor secundariorum muscle preparation of 2- to 35-day-old chicks. The nerve-induced muscle contraction was not affected by atropine and neostigmine after hatch but was slightly depressed by phentolamine and completely depressed by guanethidine. Since the responsiveness of the muscle to some adrenomimetic drugs remained constant after hatch, adrenergic receptor function does not change during development. The nerve-induced muscle response was scarcely influenced by cocaine or reserpine in neonatal chick while it was potentiated by cocaine and changed by reserpine in mature chicks. These results suggest that cocaine- and reserpine-sensitive mechanisms gradually develop in sympathetic nerve function after hatch. The presence of a potentiating effect of cocaine on the nerve-induced muscle response in dimethylphenylpiperazinium-treated neonatal chick suggests that the development of sympathetic nerve function is regulated by nerve activity itself.


Enkephalin-induced dilatation of pial arteries in vitro probably mediated by opiate receptors. Feline pial arteries in an actively contracted state were dilated in a dose dependent manner by enkephalins and morphine in vitro. This vasodilatation was blocked by naloxone suggesting the presence of opiate receptors in the vessel walls.


Effects of SQ 14,225, an orally active inhibitor of angiotensin-converting enzyme on blood pressure, heart rate and plasma renin activity of conscious normotensive dogs. Oral administration of SQ 14,225 (0.03--3 mg/kg) to conscious normotensive dogs caused inhibition of the pressor response to intravenously administered angiotensin I (AI), the duration of which was dose-dependent. All doses of 0.1 mg/kg or greater caused 85--95% inhibition 30 min after administration whereas 0.03 mg/kg produced only a 25% inhibition. Pressor responses to angiotensin II (AII) were not similarly inhibited. Blood pressure was moderately reduced in a dose-related manner and followed the same pattern as inhibition of the AI pressor responses. The maximum change occurred after 1.0 mg/kg with only a more rapid onset occurring after the 3.0 mg/kg dose. Heart rate was not appreciably changed. SQ 14,225 also increased plasma renin activity (PRA), the levels and duration of which were dose-related. These data indicate that SQ 14,225 is an orally effective, potent inhibitor of angiotensin I-converting enzyme (ACE) in dogs. It appears that in mongrel dogs, ACE inhibition results in a slight to moderate reduction in blood pressure and an increase in PRA.


Chronic antihypertensive effects of captopril (SQ 14,225), an orally active angiotensin I-converting enzyme inhibitor, in conscious 2-kidney renal hypertensive rats. Indirect systolic blood pressure (SBP) was monitored in 9 groups of 15 male conscious 2-kidney renal hypertensive rats (RHR) for over 6 months. Daily oral dosing with captopril (SQ 14,225, D-3-mercapto-2-methylpropanoyl-L-proline, 30 mg/kg), an orally active angiotensin I-converting enzyme inhibitor, lowered SBP 30--50 MM Hg during this period. Withdrawal of captopril for 5 days at 1, 3 and 6 months resulted in gradual return of SBP to control levels without overshoot. Resumption of dosage with captopril again decreased SBP. Daily oral dosing with hydrochlorothiazide (HCTZ, 6 mg/kg/day) alone for 6 months had little or no effect on SBP, but increased the antihypertensive effect of captopril. Daily oral dosing with hydralazine (6 mg/kg) caused an initial marked antihypertensive effect greater than that of captopril but almost complete tolerance developed within 4 weeks of dosing. Highest survival rates occurred in RHR treated with captopril plus HCTZ. In four other similarly treated groups of RHR and normotensive rats (NR), least cardiac hypertrophy and highest plasma renin activity occurred in captopril-treated animals compared with vehicle-treated controls. Plasma renin activity was about 2 to 4 fold higher in the rats dosed with captopril compared with vehicle-treated rats. Heart weight/body weight ratios, initially higher in the two RHR groups compared to NR, decreased only in the captopril treated group to or near those of the NR groups. These results indicate that chronic treatment with captopril decreased SBP and cardiac weights of RHR, and that HCTZ, or possibly other diuretics, can augment the antihypertensive effect of captopril while having little or no effect by themselves.


Cerebral ventricular infusion of excess calcium in the rat: effects on sleep states, behavior and cortical EEG. Sleep and behavior of the rat were recorded during cerebroventricular infusion of artificial cerebrospinal fluid (aCSF) containing regular or excess concentrations of calcium. Three different types of aCSF were used for control infusions. Depending on the ionic composition, paradoxical sleep (PS) was reduced by 6--52% during a 1-h aCSF infusion period, whereas the total amount of sleep was not altered. The depression of PS by aCSF could be prevented by increasing the concentration of Ca in the infusate by a factor of 2--5 (2.6--9.1 mM). Infusions of high concentrations of Ca (9.1--54.6 mM) caused feeding and wet-dog shakes. A slow-wave cortical EEG pattern prevailed during feeding elicited by either infusion of excess Ca or systemic administration of a small dose of pentobarbital. It is concluded (a) that the ionic composition of the CSF may selectively influence the occurrence of a sleep state, and (b) that Ca-induced feeding may be related to a covert sedative action of this cation.


Negative chronotropic action of cyclic AMP on the fetal rat heart in organ culture. Dibutyryl cyclic AMP and cyclic AMP in a DMSO medium were administered to the fetal rat heart in organ culture. Changes in heart rate were observed. Dibutyryl cyclic AMP in concentrations of 5 X 10(-5) M and 1 X 10(-4) M produced significant negative chronotropism. At a concentration of 1 X 10(-3) M, cyclic AMP in a 3% or 0.5% DMSO medium produced significant negative chronotropism. These data were compared with other studies. The negative chronotropic action of cyclic AMP and its dibutyryl derivative are evidence against the involvement of cyclic AMP in positive cardiac chronotropism.


Some clinical and epizootological observations of infectious sialoadenitis in rats. Epizootological and clinical observation was made of infectious sialoadenitis ocurring at a rat facility. The disease occurred mostly among animals 8 to 64 weeks of age without specific relevance with sex and age, and clinical signs disappeared within 5 days after the onset. Morbidity rates and duration of epizootics ranged from 13 to 100% and from 5 to 32 days, respectively. Most rats having severe clinical signs of sialoadenitis showed decrease in food uptake, body weight loss and remarkable disorders of estrous cycles.


Central receptor sites for angiotensin-induced drinking: a critical review. A review of proposed sites of the dipsogenic action of angiotensin II is presented. Techniques used for such localization are critically discussed, and it is suggested that convergence of evidence from several different experimental techniques is required for localization of dipsogenic receptors. Loci suggested as such sites of action include the preoptic regions, the subfornical organ, and the tissue proximal to the optic recess of the third ventricle, including the organum vasculosum laminae terminalis. Current evidence suggests that there are at least two loci within the forebrain that possess dipsogenic receptors for angiotensin II.


Central neural pathways for angiotensin-induced thirst. Evidence is reviewed implicating the preoptic region in angiotensin-induced thirst. The most responsive area according to results obtained with behavioral, electrophysiological, and autoradiographic mapping techniques is at the caudal border of the medial preoptic region and rostral border of the anterior hypothalamus. The neural pathway from this preoptic site for angiotensin-induced thirst extends along the medial forebrain bundle through the midlateral hypothalamus to the paramedial midbrain tegmentum and to an area ventrolateral to the central gray. Lesions of this pathway in the midlateral hypothalamus and rostral midbrain significantly attenuated drinking induced by microinjections of angiotensin II into the preoptic area but did not disrupt water intake induced by microinjections of angiotensin II into the subfornical organ or cerebral ventricles. Although the efferent pathways from angiotensin-receptive sites in the subfornical organ and cerebral ventricles are unknown, it appears from these observations that the medial forebrain bundle is not involved. Lesions of the medial forebrain bundle-lateral hypothalamus also do not disrupt drinking induced by microinjections of hypertonic saline into the preoptic region although lesions placed 1 mm further lateral do. Since fat lateral hypothalamic lesions are without effect on drinking induced by centrally administered angiotensin II, this suggests that intracellular and extracellular thirst signals are subserved by separate neural pathways in the hypothalamus.


Oxygen metabolism and the microbicidal activity of macrophages. Like neutrophils, phagocytizing macrophages undergo a "respiratory burst" in which significant quantities of oxygen are drawn into the cell. The consumed oxygen is not used in oxidative phosphorylation but, rather, in the formation of superoxide anion (O2) and H2O2. These oxygen metabolites and the products of their interaction, in particular hydroxyl radical (OH), have been implicated in the killing of ingested bacteria by neutrophils. Their role in macrophage microbicidal activity has not been fully defined. However, activated macrophages, which mediate increased resistance to infection in vivo, have a markedly increased capacity to generate O2 and H2O2 in vitro when stimulated by phagocytosis or surface perturbation. The enhanced capacity of activated macrophages to generate highly reactive oxygen metabolites during phagocytosis could contribute to the improved microbicidal and tumoricidal activity of these cells.


Inhibition of ovarian luteinizing hormone (LH) and follicle-stimulating hormone receptor levels with an LH-releasing hormone agonist during the estrous cycle in the rat. A single injection of the luteinizing hormone (LH)-releasing hormone (LHRH) agonist [D-Ala6,des-Gly-NH2(10)]LHRH ethylamide to female rats on diestrus I produced a marked reduction in ovarian LH/human chorionic gonadotropin (hCG) and follicle-stimulating hormone (FSH) receptor levels, uterine weight, and plasma progesterone levels measured 2 days later on expected proestrus. A maximal inhibitory effect was seen after a dose of only 40 ng of the peptide. No consistent effect of the LHRH analog was seen on ovarian prolactin receptors. When the analog was injected on day 7 of pregnancy, the inhibition of ovarian LH/hCG receptors was of shorter duration and was much less sensitive than that in nonpregnant animals. These data indicate that ovarian LH and FSH receptors levels are highly sensitive to changes in endogenous gonadotropin secretion and suggest that the gonadotropin surge occurring spontaneously on proestrus may play an important role in the regulation of ovarian gonadotropin receptors during the estrous cycle.


Prostaglandins and opioids.

Prostaglandins and opioids. In guinea-pig small intestine, rat brain in vitro and neuroblastomaXglioma hybrid cells, opioids specifically inhibit the action of E prostaglandins. In the whole rat, E prostaglandins, administered centrally, antagonize the antinociceptive action of morphine. E prostaglandins also antangonize the induction of opioid tolerant/dependence. In turn, tolerance/dependent preparations respond with added intensity to E prostaglandins. The antagonism between opioids and E prostaglandins does not occur at the opioid receptor; but, certainly in some preparations and probably in others, this antagonism occurs at the coupling or catalytic unit of a neuronal adenylate cyclase that opioids inhibit and E prostaglandins stimulate. The proposition that antagonism of E prostaglandin at appropriate neurons in the brain is part of the natural mechanism of opioid analgesia remains possible, but unproven, and is worth continued investigation.


The effect of propranolol on hypoglycaemia. Observations in five insulinoma patients. Five hypoglycaemic hyperinsulinaemic patients (three with proven benign insulinoma, one with proven metastasizing insulinoma, one with probable insulinoma not found at surgery) were treated with propranolol for a variable time ranging from two weeks to one year. Three patients showed favourable clinical results and a significant increase of the mean basal blood glucose level was found while two patients showed no improvement of the frequency of neuroglycopenic episodes and no significant increase of their mean blood glucose level. No patient showed a significant decrease in mean basal IRI concentration. A decrease of insulinaemic responses was observed during oral and intravenous glucose tolerance tests, a prolonged fast, and tolbutamide and glucagon tests performed in some patients. The results suggest that propranolol may induce in certain patients an improvement of basal clinical status through not understood effects (probably hepatic), which leave the peripheral concentrations of insulin unchanged, whereas inhibition of insulin secretion may represent the main way by which the improvement of metabolic situation during physiological or pharmacological stimulation may have been achieved.


Relative insensitivity to glucagon of sterol synthesis in cultured rat aortic smooth muscle cells. Effect of dibutyryl cyclic AMP. The smooth muscle cell plays an important role in the process of atherogenesis. In these experiments the effect of glucagon and dibutyryl cyclic AMP on sterol synthesis in cultured rat arterial smooth muscle cells was studied. Glucagon in concentrations of 1 X 10(-9) mol/l inhibited the incorporation of sodium (2(-14)C)acetate into non-saponifiable lipids and digitonin precipitable sterols but lower concentrations of glucagon had no effect. In cells which were exposed to serum, dibutyryl cyclic AMP also resulted in a decrease in the incorporation of labelled acetate into sterols but when the cells were grown in serum free medium, dibutyryl cyclic AMP had no inhibitory effect on sterol synthesis. These results provide further evidence that sterol metabolism in arterial smooth cells may be influenced by hormones but suggest that glucagon is relatively less important than insulin in this respect.


Viral studies in streptozotocin-induced pancreatic insulitis. Multiple injections of streptozotocin to Charles River (CD-1) Laboratory mice resulted in a syndrome characterised by diabetes mellitus, insulitis and the induction of endogenous type C viruses in pancreatic beta cells. Within one week after the completion of five intraperitoneal injections of streptozotocin, the CD-1 mice exhibited irreversible hyperglycaemia and insulinopaenia. Light microscopic studies of pancreata from mice sacrificed at this time demonstrated insulitis and beta cell necrosis. Electron microscopic studies revealed spherical and atypical cylindrical type C viruses and occasional clusters of intracisternal type A viruses exclusively within beta cells. To clarify the identification of the type C viruses and their role in the genesis of the insulitis, type C virus specific antigens were identified within islet cells by immune fluorescence at various intervals after streptozotocin administration. Immune fluorescence studies demonstrated the presence of type C virus antigens within islets from streptozotocin treated mice but not in buffer-injected controls. Time course studies suggested that type C virus induction may precede the appearance of insulitis by two days and that insulitis is consistently accompanied by the presence of virus positive islet cells.


Cellular injury of adrenocorticotropic pituitary tumor (AtT-20) by corticosteroid in mice. A transplantable ACTH-secreting pituitary tumor (AtT-20) in mice was studied for its growth in relation to adrenocortical steroid. The latency of transplanted tumor in LAF-1 mice with 10(6) tumor cells was about 2 weeks in adrenalectomized and in intact mice. In the former, however, tumor size reached 1.5 cm in diameter within 1 month, while it remained about 0.5 cm in the latter. Macroscopic and microscopic observation revealed that the tumor grafted in intact mice showed massive necrosis, leaving minimum intact tumor cells among necrotic foci. In contrast, the tumor grafted in adrenalectomized mice was healthy, soft, and showed adenomatous structure. It is concluded that excess of adrenocortical steroid produced in the grafted AtT-20 was inhibitory for the growth of transplanted tumor and removal of the adrenal glands was good for growth and survival of transplanted tumor. For the assessment of direct effect of adrenal steroid on AtT-20 cells, incorporation of 3H-thymidine in cultured AtT-20 cells was examined in a medium supplemented with glucocorticoid hormone. The autoradiographic study indicated that the inhibition of 3H-thymidine uptake was demonstrated at 24 hr after incubation with steroid and labeled cells were barely detectable on 3rd and 5th day.


Occurrence of antibody against intracytoplasmic A-particles of mouse mammary tumor virus in sera from breast cancer patients. Acetone-fixed smears of DBA/2 mouse leukemia cells that produce clusters of intracytoplasmic A-particles (pronucleocapsids of mouse mammary tumor virus) were employed as an indirect immunofluorescence system to detect the antibody to A-particles in human sera. With positive test sera, specific fluorescence was easily detectable as discrete cytoplasmic granules at the site of A-particle clusters. The antibody was found in 26 (60%) out of 43 breast cancer patient sera and 4 (25%) of 16 mammary fibroadenoma patient sera, while only 4 (11%) out of 37 control woman sera were antibody-positive. In the case of breast cancer patients, occurrence of tha antibody was not specifically related to a particular type of tumor histology. In a considerable number of positive cases, the antibody tended to disappear within various lengths of time after surgical operation of the breast cancer.


Phosphorylation of 1-beta-D-arabinofuranosylcytosine by the cell-free extract of rat ascites hepatoma, in relation to the mechanism of natural resistance. The enzyme that phosphorylates 1-beta-D-arabinofuranosylcytosine (ara-C) was examined in rat ascites hepatomas to clarify whether the capacity for the phosphorylation of ara-C would be related to natural resistance of the tumors to ara-C. Ara-C kinase activity in AH-66F cells, which were moderately sensitive to ara-C, was approximately the same as that in the bone marrow of tumor-bearing rats. L-1210 leukemia, which is sensitive to ara-C, had a higher activity of the enzyme than the mouse bone marrow. On the other hand, the naturally resistant hepatomas, AH-60C, AH-109A, and AH-66, were low in their ara-C kinase activity. The enzyme activity in the cytoplasmic extract (30,000 g supernatant) of the tumor and bone marrow cells was nearly proportional to the capacity for ara-C phosphorylation in intact cells. Thus, the mechanism of natural resistance to ara-C in rat ascites hepatomas was attributed mainly to the low levels of ara-C kinase activity itself. Apparent Km of the enzyme for ara-C was about 170 micron. However, probably due to the presence of the uncompetitive type of inhibitor(s), the level of apparent Km lowered close to 40 micron at higher concentration of the protein as the source of the enzyme, showing similarity to the value in intact cells.


Clinicopathological study on 100 early gastric cancer cases. Clinicopathological study was given to 100 early gastric cancer cases. The cases included 50 intramucosal lesions (m-group) and 62 submucosal lesions (sm-group), and the share is 17% of whole cases treated surgically. The sex ratio is about 2:1 (male : female). According to the classification by location, the occurrence rate of lesion on anterior wall is 18.8%. This fact suggests the impotance of roentgenscopy by compression technique and double contrast in a prone position focussed on anterior wall. In addition, endoscopy is essential in order to detect minute cancers. The rates of metastasis of early gastric cancers is 2.4% of m-group and 16.1% of sm-group. No metastasis occurs in protruded or elevated type as far as cancer cells remain within the mucosa. Once cancer cells infiltrate into the submucosa, metastasis is observed in 36%. In the present cases, 9 (9%) out of 100 cases of early gastric cancers are the multiple cases. In order to avoid oversight of cancer foci, the portion of stomach to be remained after surgery should be throughly examined prior to operation and again under direct vision after gastric incision.


Serum alkaline phosphatase (Al-Pase) isozyme in gastric and colonic cancer (using a simple thin layer polyacrylamide gel electrophoresis). Using the simple thin layer polyacrylamide gel electrophoresis, serum alkaline phosphatase could be separated 5 isozyme bands in various digestive diseases, consisting of 54 cases of gastric cancer, 11 of colonic cancer, 12 of hepatoma, 4 of cholangioma, 14 of pancreatic cancer, 81 of benign hepatobilliary diseases, 13 of cancers of other organs and 61 of control. The obtained results were as follows: 1) The electrophoretic analysis of serum alkaline phosphatase showed the specific band remaining at the origin, already reported as "alkaline phosphatase O", in primary and metastatic cancer of the liver and cholelithiasis. On the contrary, alkaline phosphatase O was never found in gastric and colonic cancer without cholelithiasis. On the contrary, alkaline phosphatase O was never found in gastric and colonic cancer without cancerous metastasis to the liver, and it was also inclined to be positive with the progress of liver metastasis among them. 2) Intestinal alkaline phosphatase was usually found in higher frequency in blood group B and O than in the others, and it was apt to disappear in gastric or colonic cancer with an exacerbation of its cancerous lesions. 3) Heat-stable alkaline phosphatase was found in 10% of gastric or colonic cancer, all of which were histologically proved to be well differentiated adenocarcinoma.


Cyclic AMP metabolism and adenylate cyclase concentration in patients with advanced hepatic cirrhosis. Glucagon was tested for its effect on plasma adenosine 3',5'-cyclic monophosphate (cyclic AMP), insulin, and glucose in healthy subjects and in patients with advanced cirrhosis of the liver. In the normal subjects, intravenous infusion of glucagon caused a significant increase in plasma cyclic AMP, glucose, and insulin. In advanced cirrhotics, plasma cyclic AMP, glucose, and insulin did not increase. Adenylate cyclase concentration was measured in liver tissue from end stage cirrhotic patients and from brain-dead organ donors whose cardiovascular function was maintained in a stable state. Basal and total adenylate cyclase concentration were not different in the two groups. Adenylate cyclase from the livers of advanced cirrhotics was, however, significantly less responsive to glucagon stimulation than was that from donor livers. Hepatocytes in advanced cirrhosis have abnormal metabolic behavior characterized by abnormal adenylate cyclase-cyclic AMP response to hormonal stimulation.


Perhexiline maleate-induced cirrhosis.

Perhexiline maleate-induced cirrhosis. The authors report the cases of 2 patients who died from cirrhosis after receiving perhexiline maleate, a drug widely used in Europe for the treatment of angina pectoris. Perhexiline maleate had been ingested for 24 and 28 mo, respectively. Manifestations of cirrhosis included jaundice, hepatic encephalopathy, ascites, and portal hypertension. Associated manifestations of intolerance to perhexiline maleate included peripheral neuropathy in 1 patient and marked weight loss in both. Histologic lesions resembled those observed in patients with alcoholic liver disease. Ultrastructural lesions included numerous enlarged lysosomes containing myeloid figures. Histochemical stains demonstrated increased phospholipid content of the hepatocytes. These findings are consistent with the view that prolonged administration of perhexiline maleate may induce both histologic lesions resembling those of alcoholic liver disease and ultrastructural and histochemical lesions resembling those of phospholipidosis.


Ulnar neuropathies in rheumatoid arthritis. A loss of functional motor axons in the median and ulnar nerves occurred in half of thirty-three patients with rheumatoid arthritis. Weakness of small hand muscles may predispose to the development of ulnar deviation of the fingers in patients with joint disease at the radio-ulnar and metacarpophalangeal joints. There is no evidence that spasm of small hand muscles is a significant cause of ulnar deviation of the fingers in rheumatoid arthritis. Ulnar deviation of the fingers in rheumatoid arthritis is not due to selective impairment of the ulnar nerve or the deep palmar branch of the ulnar nerve even though ulnar deviation of the fingers can occur in association with such lesions and in the absence of joint disease.


Syndactyly and split hand. Supplement.

Syndactyly and split hand. Supplement. In a series of studies of clinical and radiological features of hand anomalies, the author has recently encountered cases which may be explained by reference to the relationship between syndactyly and split hand. These cases suggested how the typical split hand is formed prenatally, and it is concluded that the typical split hand should not be classed as "Arrest of Development of Parts" due to embryonal failures.


[Cervix carcinoma as an immunological model. Part 2. In vitro studies and HLA system typing]. Carcinoma of the cervix uteri is one of the most important tumors in women. The tumor induces an immunological defense reaction against itself in the tumor host. The round cell infiltrate at the tumor invasion line, which predominantly consists of small lymphocytes is the morphological equivalent of this interrelationship. In the lymph-nodes of the drainage area of the tumor certain patterns of reaction can be differentiated, which may be interpreted as humoral or cellular immune reactions. Cellular immune reactions in lymph-nodes of patients operated according to the method of Wertheim-Meigs were correlated with a better prognosis of the tumor disease. The hypothesis of an involvement of the HSV-type II in the genesis of the cervical carcinoma is supported by the increased incidence of antibodies against the virus. Therewith, relations between the stage of disease and the age of the patient were found as a possible basis for the virus hypothesis and the immune reaction to the tumor associated antigens an altered genetic situation may be presumed. Typing in the HLA-system demonstrates an accumulation of the frequency of the antigen B12 in patients with carcinoma of the cervix. These investigations may lead to a differentiated outlook with regard to the detection of risk groups and therapeutical consequences for cervical cancer.


Parathyroid hormone and triglyceride transport: effects on triglyceride secretion rates and adipose tissue lipoprotein lipase in the rat. In order to determine the effects of parathyroid hormone on triglyceride transport, male Sprague-Dawley rats were injected with parathyroid extract for eight days and triglyceride secretion rates (TGSR) and adipose tissue lipoprotein lipase (LPL) activity were determined. Parathyroid hormone-treated rats demonstrated significantly lower (p less than .005) TGSR in the basal overnight fasted state 15 hours after the previous injection, but this effect on TGSR was not apparent 3 hours after PTE injection when the rats were allowed to eat. In contrast, LPL activity was significantly reduced in the PTH-treated animals at 3 hours and no effect was apparent on this index of triglyceride removal at 15 hours. These findings suggest that parathyroid hormone may exert independent influences on triglyceride production and removal and thus may alter triglyceride homeostasis in conditions in which parathyroid hormone levels are abnormally increased.


Studies on cytochrome c oxidase, II. The chemical constitution of a short polypeptide from the beef heart enzyme. A low molecular weight (approximately 6000) polypeptide fraction was isolated from beef heart cytochrome c oxidase, consisting of three peptides with the N-terminal end groups isoleucine, phenylalanine and serine. The complete amino acid sequence of the serine component is described. From the chemical constitution, a site-specific cleavage from a precursor protein and a possible function in membrane penetration and complex formation of the oxidase is inferred.


Superoxide-independent platelet response to xanthine oxidase. Xanthine oxidase (1--5 microgram/ml) from cow's milk induces shape change, aggregation, and the release reaction of human washed platelets. Xanthine oxidase plus xanthine produce superoxide radicals, which reduce nitro blue tetrazolium. Superoxide dismutase, allopurinol, or ommission of xanthine inhibits the reduction of nitro blue tetrazolium but has no influence on the platelet response to xanthine oxidase. In contrast, small amounts of plasma or apyrase from potatoes abolish the effect on platelets, but not the enzyme activity of xanthine oxidase. Comparison of two xanthine oxidase preparations shows that higher specific enzyme activity corresponds to a lesser effect on platelets. The results suggest that platelet and enzyme activities reside in different components of xanthine oxidase preparations.


A social interaction program for chronic psychiatric patients living in a community residence. A psychiatric center in Brooklyn provided a socialization program in a community residence for adults discharged from state hospitals and made a study of the program's outcome. Residents on two floors of the home, the experimental group, received an enriched treatment program; those on the other four floors, the control group, received a minimum of therapeutic intervention. A comparison of residents' ratings on assessment scales made early in the program and after seven months of operation showed that residents on all floors improved on measures of socialization, men improved more than women, and residents over 65 years of age improved the least. Residents in the experimental group did not improve much more than those in the control group. Costs for the experimental group, including room and board, were $16.59 a day per resident. Costs for the control group were $15.61 a day per resident.


Leukocytes as secretory organs of inflammation. Polymorphonuclear leukocytes secrete substances that defend against foreign invaders by mechanisms that bear remarkable resemblances to those of exocrine and endocrine glands. Similarly, leukocyte secretions can also damage the very host they are intended to protect. Like other secretory cells, leukocytes also bear surface receptors to detect signals that initiate secretion. Clinical implications are detailed.


The kidney in health and disease: IV. Principles of ion and water transport in the kidney. Modifications within tubular cells facilitate both outward movement and reabsorption of water and solutes through the lipid bilayer plasma membrane. The membrane transport systems that regulate these movements involve processes that obey the laws governing membrane transport in all biologic systems. Evidence that independent pathways for water and for certain ions may exist in cell membranes is discussed.


Ultrastructural and biochemical analysis of "undifferentiated" pulmonary carcinomas. Seven cases of "undifferentiated" pulmonary carcinoma were studied ultrastructurally; five were of the typical oat cell variety and the remaining two consisted of larger cells. In three of the former and both of the latter cases neurosecretory-like granules were demonstrated. Biochemical analysis of tumor tissue extracts revealed 5-hydroxy-3-indoleacetic acid, vanilylmandelic acid, and catecholamine activity in all instances. No hormonal syndrome or metabolic abnormality was detected in any of the patients. The concomitant morphologic demonstration of neurosecretory-like granules and the presence of 5-hydroxy-3-indoleacetic acid, vanilylmandelic acid, and catecholamines in neoplastic tissue would provide further evidence that these tumors may indeed arise from bronchial endocrine cells and could therefore be classified within the group of neuroendocrine carcinomas. Also it seems apparent that these neuroendocrine bronchial carcinomas may include tumors consisting of cells somewhat larger than the typical oat cell. The observation of 5-hydroxy-3-indoleacetic acid, vanilylmandelic acid, and catecholamine activity in two oat cell carcinomas in which neurosecretory granules could not be demonstrated poses an interesting problem whose solution may only derive from further studies.


The role of electron microscopy in the diagnosis of unusual peripheral lung tumors. The ultrastructure of the lung provides a cytomorphologic basis for the identification of unusual pulmonary neoplasms or unusual histologic variants of more common pulmonary lesions. Comparison of tumor cells with bronchioloalveolar lining cells and with pleural components has aided in the diagnosis of a spindle cell variant of a peripheral neuroendocrine cell tumor (carcinoid) and a tumor composed of cells resembling bronchioloalveolar epithelium (sclerosing hemangioma of lung).


Leukotic spread: an unusual tumor transition. This report concerns a case of lung cancer, which is of interest because it proves that an epithelial tumor is capable of spreading in a manner morphologically identical to that of a malignant lymphoreticular tumor.


Lysosomal acid hydrolases in established lymphoblastoid cell lines, transformed by Epstein-Barr virus, from patients with genetic lysosomal storage diseases. Lysosomal acid hydrolases were determined in established lymphoblastoid cell lines, transformed in vitro by Epstein-Barr virus (EBV) from lymphocyte-rich cell populations isolated from the peripheral blood of patients with genetic lysosomal storage diseases--Hurler syndrome, Scheie syndrome, GM1-gangliosidosis type 1 and type 2, Tay-Sachs disease, and I-cell disease--and from obligate heterozygotes for these diseases. The respective enzyme activity was undectectable in lymphoblastoid cells from the patients, but not from controls. Obligate heterozygotes could not always be distinguished from controls in lymphoblastoid cells as well as in leukocytes. These results suggest that established lymphoblastoid cell lines are useful material for the enzymatic study of genetic lysosomal storage diseases.


Superoxide production from human polymorphonuclear leucocytes stimulated with immunoglobulins of different classes and fragments of IgG bound to polystyrene dishes. Polystyrene surfaces coated with proteins at alkaline pH were found to be useful to investigate the stimulation of neutrophils independently of phagocytosis. Human neutrophils, when exposed to appropriate stimuli, release superoxide anion (O2-). We have measured superoxide dismutase-inhibitable cytochrome c reduction by cells stimulated with various kinds of protein coated on polystyrene dishes. Native immunoglobulins in solution did not stimulate O2- generation in neutrophils. IgG and IgA adherent to polystyrene dishes stimulated O2- generation in neutrophils, but IgM, IgD, IgE, albumin, Fab, and Fc of IgG on polystyrene dishes did not.


Persistent shedding of adenovirus in urine of chimpanzees. A new adenovirus, designated Pan 11, was isolated repeatedly from the urine of several chimpanzees for more than 1 year. One chimpanzee had chronic interstitial nephritis; the others were healthy. Most chimpanzees tested had neutralizing antibodies to Pan 11 virus in the serum; three people who worked with chimpanzees also acquired antibodies. Transplantable rhabdomyosarcomas developed in hamsters inoculated as newborns with Pan 11 virus.


Analysis of antigenic diversity among human cytomegaloviruses by kinetic neutralization tests with high-titered rabbit antisera. Neutralizing antisera to human cytomegalovirus were produced in rabbits with alkaline-buffered extracts of infected cell cultures. The antibody activity was complement dependent and associated primarily with the 7S immunoglobulin fraction. Antisera with homologous K values greater than 10.00 were shown to be suitable for neutralization kinetic studies and were so used to examine the antigenic relatedness of strains AD169, Davis, Esp, C-87, Kerr, and Towne. Based upon the degree of relationship as determined by normalized K values, antisera to the Davis or AD169 strains discriminated three antigenic groups, and an antiserum to strain Esp discriminated two groups.


Interstitial pneumonia and subclinical infection after intranasal inoculation of murine cytomegalovirus. Although cytomegalovirus (CMV) infections are common throughout the world, little is known about the means of person-to-person transmission. To determine whether infection could be established by a respiratory route, studies were conducted in a murine CMV (MCMV) model by using intranasal inoculation. The infectious dose which resulted in pulmonary and systemic infection of half the mice was 100 plaque-forming units of MCMV. Here, infection was subclinical, but virus replicated in the lungs and subsequently disseminated via the blood to other organs within 7 days. The serum immunofluorescence antibody titer peaked by day 21. None of these mice died, although focal peribronchial interstitial pneumonitis was found in infected animals. In mice given greater than or equal to 10(4) plaque-forming units of MCMV intranasally, severe diffuse interstitial pneumonitis resulted uniformly, closely resembling that seen in immunocompromised patients and in newborn infants, and 20% of the animals died. Normal pulmonary architecture was obliterated by sheets of histiocytes, many containing MCMV intranuclear inclusions, and by accumulation of proteinaceous fluid in the interstitial and alveolar spaces. Of relevance to human disease, these experiments show that MCMV as a sole pathogen can cause severe interstitial pneumonitis in normal mice and that subclinical systemic infection results from respiratory inoculation of small amounts of virus.


Ultracentrifugal inoculation of herpes simplex virus. By ultracentrifugation of 30 ml of highly dilute suspensions of herpes simplex virus (HSV) directly onto monolayer cultures grown in centrifuge tubes, infectivity was significantly greater than without centrifugation. Ultracentrifugation at 20,000 to 25,000 rpm (28,000 to 45,000 X g) for 1.5 to 2.3 h was utilized with good preservation of cultures. With low-speed centrifugation at 3,000 rpm (1,100 X g), infectivity was almost 10-fold greater than without centrifugation. With ultracentrifugal inoculation, infectivity was about 100-fold greater than without centrifugation. Ultracentrifugal inoculation permitted the detection of HSV at concentrations as low as 0.05 plaque-forming units per ml. Similarly, ultracentrifugal inoculation of cultures was almost 100-fold more sensitive a method of detecting infectious HSV than was pelleting HSV from dilute suspensions followed by resuspension and inoculation of cultures. Ultracentrifugal inoculation of cultures may permit the isolation of HSV in situations where virus cannot be detected by ordinary means and may prove applicable to the study of other viruses.


Lymphocyte-mediated cytotoxicity in humans during revaccination with vaccinia virus. Fifteen healthy human volunteers were revaccinated with vaccinia virus. Blood samples (4 to 7) were obtained during the 3 weeks after revaccination. Peripheral blood lymphocytes were washed extensively and tested for cytotoxicity against vaccinia-infected autologous and/or homologous skin fibroblasts. Without addition of antibodies, peak levels of killing were observed on days 7 to 9. The killing did not depend on common HLA markers. On days with peak activity, extensively washed lymphocytes showed higher levels of killing than normally washed lymphocytes. By cell separation experiments, the cell most active in killing proved to be a nonadherent, non-phagocytizing lymphocyte with Fc receptors. Serum antibodies tested in two sensitive serological assays peaked on days 14 to 17. The question of whether the killing observed is dependent on or independent of antibodies is not clarified in the present study.


Effects of ambient temperatures on induction of transmissible gastroenteritis in feeder pigs. Experiments were carried out to investigate the effects of ambient temperatures on the induction of transmissible gastroenteritis in feeder pigs 2 to 3 months old. Pigs maintained at a high temperature (30 +/- 2 degrees C) and exposed to the virulent transmissible gastroenteritis virus did not show clinical signs of the disease during their maintenance at the high temperature. On the other hand, a sudden decrease in the ambient temperature, either before or after virus inoculation, induced severe disease in feeder pigs exposed to the virus. However, continuous maintenance of pigs at the low temperature (4 +/- 1 degrees C) tended to somewhat reduce the frequency of occurrence of signs in proportion to the length of the maintenance periods at that temperature. Pigs raised at temperatures that fluctuated between 20 +/- 2 and 4 +/- 1 degrees C every 24 h developed profuse diarrhea. The duration of clinical signs was longer in pigs maintained under the fluctuating temperatures than in those at the constantly low temperature. With one exception, antibody against transmissible gastroenteritis virus was demonstrated in sera collected from pigs both with and without clinical signs. Antibody titers obtained, however, were somewhat higher in sera collected from pigs that had developed clinical signs than in those from pigs that had endured the infection without showing signs.


Role of hydrogen peroxide and peroxidase in the cytotoxicity of Trypanosoma dionisii by human granulocytes. The mechanism of the cytotoxic reaction of leukocytes to Trypanosoma dionisii was investigated. Cytotoxicity was measured by release of [99mTc]pertechnetate from labeled protozoa. Both granulocytes and lymphocytes were found to be cytotoxic to antibody-coated T. dionisii. The reaction was inhibited by diethyldithiocarbamate and by potassium cyanide, both of which inhibit myeloperoxidase. Myeloperoxidase from azurophil granules was toxic to T. dionisii, provided that hydrogen peroxide was also present. Hydrogen peroxide formation was induced in granulocytes and, to a lesser extent, in lymphocytes by antibody-coated T. dionisii. Inhibition of this hydrogen peroxide formation by treatment of the effector cell surface with p-diazobenzenesulfonic acid inhibited cytotoxicity. It is therefore concluded that granulocytes, and probably also lymphocytes, kill T. dionisii with hydrogen peroxide by a peroxidase-mediated reaction. Although hydrogen peroxide and myeloperoxidase alone were also cytotoxic to the lymphoblastoid cell line CLA4, it seems unlikely that this is the cytotoxic mechanism for this process because these cells were unable to induce hydrogen peroxide formation.


Protective effect of early serum from mice after cytomegalovirus infection. Serum obtained from mice 3 to 5 days after cytomegalovirus infection contained complement-requiring neutralizing antibody, which, when passively transferred, protected mice from lethal infection with the homologous virus. The active substance was largely heat-stable 7s immunoglobulin.


Glycoproteins of natural origin with an affinity for hepatitis B surface antigen. Sera from certain animal species contain a substance(s) which binds hepatitis B surface antigen. The hepatitis B binding substance found in animals is not antibody, but appears to be a glycoprotein which reacted with antigen-coated beads and produced a "false positive" test for antibody. This glycoprotein could be selectively and quantitatively removed by reaction with purified hepatitis B surface antigen and centrifugation. Pili fractions isolated from Neisseria gonorrhoeae and Escherichia coli bound to hepatitis B surface antigen and produced false positive anti-hepatitis B surface antigen reactions. Mouse anti-bovine hepatitis B binding substance and rabbit anti-E. coli pili were capable of neutralizing bovine hepatitis B binding substance.


Peritoneal macrophage activation indicated by enhanced chemiluminescence. A number of studies have demonstrated the ability of various bacterial preparations, protozoa, and chemicals to activate macrophages and concomitantly to enhance host resistance to both tumors and infections. Recently, viral infections have been shown to have a similar effect upon macrophage function. To better define the metabolic state of activated macrophages, we have evaluated the ability of peritoneal cells (PC) from vaccinia virus- or murine cytomegalovirus-infected or Corynebacterium parvum-treated mice to emit chemiluminescence (CL) during phagocytosis of zymosan particles or yeasts. PC from C. parvum-treated mice (1,400 microgram intraperitoneally) emitted enhanced CL over controls on days 3, 6, 14, and 21 after treatment, thereby establishing the emission of CL as a correlate of metabolic activation. Previous evidence for activation of PC from vaccinia virus-infected mice (10(8) plaque-forming units) was confirmed by demonstration of enhanced levels of CL on days 3, 6, and 13 after murine infection. Likewise, PC from mice infected with murine cytomegalovirus (10(5) plaque-forming units) 3, 6, or 13 days previously demonstrated augmented levels of CL over controls. Opsonized virus particles (vaccinia virus or murine cytomegalovirus) failed to induce the emission of CL with PC from mice infected with the isologous virus. Our data further demonstrate the immunomodulationinduced by virus infections and suggest that the detection of CL is an easily quantitated correlate of macrophage activation which may be helpful in defining metabolic alterations induced during activation.


Interferon treatment of NZB mice: accelerated progression of autoimmune disease. The effect of long-term administration of interferon in New Zealand Black and New Zealand Black/New Zealand White F1 hybrid mice was studied. Treatment with moderate doses of interferon (10(4) units, five times weekly for 8 weeks) did not depress murine leukemia virus gp69/71 levels in serum and spleen, nor p30 levels in the spleen. Interferon given at 10(5.1) units (three times weekly for 37 weeks) caused an increased incidence of anti-erythrocyte antibodies in New Zealand Black mice. Finally, the hybrid mice given interferon at 10(6.0) units (three times weekly for 33 weeks) had increased renal immune complex deposits and increased incidences of proteinuria and anemia.


Some aspects of the physiological and pharmacological control of the synthesis of triacylglycerols and phospholipids. The effects of dietary modification, starvation, stress and diabetes on the activity of phosphatidate phosphohydrolase are discussed. Evidence is presented that this enzyme is involved in controlling the rate of triacylglycerol synthesis in the liver. Drugs such as fenfluramine and benfluorex are able to inhibit phosphatidate phosphohydrolase by interacting with the substrate. This decreases the rate of triacylglycerol synthesis and redirects the route of glycerolipid metabolism. Benfluroex also partly prevents the ethanol-induced increase in triacylglycerol synthesis and in phosphohydrolase activity. The implications of these findings are discussed with respect to the mode of action of fenfluramine and benfluorex and to the control of triacylglycerol synthesis.


The synthesis of N-benzyloxycarbonl-L-prolyl-dehydrophenylalanyl-L-histidyl-L-leucine: a converting-enzyme inhibitor. The direct oxidation of a dipeptide azlactone (1) by DDG affords an optically active unsaturated dipeptide azlactone (2). It was shown that the double bond had the Z-configuration and that no racemization of the proline moiety occurred during the oxidation. The dehydrotripeptide, Z-Pro-delta-Phe-Phe OH (9b) was prepared by direct coupling of azlactone (2) with phenylalanine tetramethyl-guanidinium salt and was shown to be stable to chymotrypsin. The tetrapeptide, Z-Pro-delta-Phe-His-Leu  OH (11), was prepared directly from the azlactone (2) and by saponification of the tetrapeptide ester (10) prepared by mixed anhydride coupling of Z-Pro-deltaPhe  OH (7) with the dipeptide ester. The dehydropeptide (11) inhibits angiotensin I converting enzyme (IC50, 1X10-4 M).


The effect of cesium salts on dense poliovirus particles. The buoyant density of dense poliovirus particles has been examined in density gradients other than cesium chloride in order to determine the dependence of this property on the nature of the solvent. In Urografin (sodium and methylglucamine amidotrizoate), dense poliovirus particles banded at two densities--1.33 and 1.39 g/cm(3)--whereas in cesium metrizoate they banded only at 1.39 g/cm(3) and in cesium sulfate at 1.38 g/cm(3). The presence of cesium ions gives rise to the occurrence of dense particles, which are indistinguishable in their buoyant density and sedimentation coefficient from standard poliovirus particles when prepared in the absence of Cs+. In their physical properties dense poliovirus particles are more closely related to rhinoviruses and foot-and-mouth-disease virus, i.e., to members of the genus Rhinovirus, than to standard poliovirus particles belonging to the genus Enterovirus.


Insulin inhibits cellular and herpes simplex virus DNA synthesis in transformed lymphoblastoid cell lines. Treatment of uninfected and herpes simplex virus-infected lymphoblastoid cells with 4 units of insulin per milliliter resulted in the inhibition of both cellular and viral DNA synthesis.


[Liver resection in benign and malignant tumors]. Benign tumors of the liver, circumscribed hepatomas and solitary hepatic metastases from colonic cancer are treated by partial liver resection. In case of colonic cancer the hepatic metastasis is resected in a second operation. 20 cases of hepatic resections are reported. 10 right hepatic lobectomies, 1 left hepatic lobectomy and 9 minor resections were performed. Of the 11 patients treated by hepatic lobectomy, one (or 9%) died in the postoperative period (within 30 days after operation). Of the other 9 patients undergoing minor resections there was no death.


Exhaustive physical exercise and acid hydrolase activity in mouse skeletal muscle. A histochemical study. Adult, untrained NMRI mice were exhausted on a motor-driven treadmill by an intermittent-type running programme. Serial cryostate sections for the staining of NADH-tetrazolium reductase, beta-glucuronidase, beta-N-acetylglucosaminidase, and beta-glycerophosphatase activities and for making hematoxylin-eosin staining were cut from m. quadriceps femoris 1, 2, 3, 5, 7, and 15 days after physical exhaustion. A strong increase in the activities of beta-glucuronidase and beta-N-acetylglucosaminidase was observed 7 days after exhaustion and the activity changes, which were similar for the both glycosidases, were more prominent in the highly oxidative red compared to less oxidative white fibres. Activity granules were more numerous in the perinuclear than the interfibrillar area of red fibres. Spots were arranged like longitudinal chains between myofibrils. Activity in connective tissue was usually observed only in animals exhausted 3--7 days earlier. Simultaneous activity in fibres exceeded that in connective tissue. beta-Glycerophosphatase activity was not, by the method used, seen in histologically "healthy" or normal-looking fibres. In samples taken 2--5 days after exhaustion some degenerating and necrotic fibres were observed. Inflammatory reaction was also observed being at its strongest five days after loading when mononuclear cells were seen inside necrotic fibres. The number of regenerating muscle cells was most abundant 7 days after exhaustion. It is suggested that temporary hypoxia, which accompanies exhaustive physical exercise in skeletal muscle, upsets the energy metabolism and homeostasis of fibres and causes the observed histological and histochemical alterations, which possess features typical of both lethal and sublethal acute cell injury.


Lung tissue guanosine 3',5'-monophosphate: effects of ventilation and anesthesia. In vivo levels of guanosine 3',5'-cyclic monophosphate (cGMP) in rat lung were measured by radioimmunoassay. The basal levels of this agent were found to change depending on the method of killing or anesthesia used. Lowest values were found after microwave irradiation to the head (10.2 fmol/mg wet wt). Values after pentobarbitol anesthesia, with or without atropine, were significantly higher (19-24 fmol/mg). In animals anesthetized with pentobarbital and ventilated at pressures of 7 cmH2O with no end-expiratory pressure, lung cGMP was 42-49 fmol/mg. Ventilation at higher pressures resulted in much higher levels of cGMP (20 cmH2O, 105 fmol/mg; 30 cmH2O, 233 fmol/mg). The response of cGMP to ventilation was not blocked by pretreatment of the animals with atropine. High-pressure ventilation was without effect on lung adenosine 3',5'-cyclic monophosphate.