An analysis of distance variables that affect aftercare attendance.

Recent reserach has examined variables related to attendance of aftercare services because of the correlation between aftercare and posthospitalization treatment success. The present study, which examined aftercare attendance for 40 randomyl selected subjects following discharge from an inpatient alcohol treatment program, extends the list of variables found to influence attendance. A multiple-regression analysis of subject demographic variables and variables of a more programmatic nature found that distance variables (e.g., number of miles to an aftercare agency) significantly influenced aftercare attendance. The results are discussed with respect to aftercare program decisions.


Simulation of coupling between chemical reactions and ion transport in brown adipose tissue using network thermodynamics.

Several cellular events associated with energy turnover in the mitochondria and at the Na+/K+ plasma membrane pump have been formulated in terms of network thermodynamics. The calorigenic role of the Na+/K+ pump is examined in terms of the relationship between the movement of sodium and potassium ions and the chemical reactions involved. In addition, attention is centered on the potential thermogenic role of three mitochondrial pathways involving proton fluxes--namely, one in which protons are transported from the matrix to the intermembrane space; a second in which protons are transferred back into the matrix in conjunction with the synthesis of ATP; and a third wherein protons re-enter the matrix without being coupled to any chemical reaction. (This latter pathway has been delineated by studies on isolated mitochondria and may be unique to brown fat.) At both sites (plasma membrane and mitochondria) the conversion of chemical energy to heat is considered.


Characterization of increased synthesis of acute phase proteins from plasma concentration measurements: a system analysis of the ideal situation with use of computer simulation.

A method for characterizing the secretion of acute phase proteins in vivo from serial plasma concentration measurements is presented. By using a simple two-compartment model it was found that descriptive parameters could be derived from the time course of the increased plasma concentration of the protein. It was assumed that the secretion of an acute phase protein is adequately described by: (i) the start time t1; (ii) the end time t2; (iii) the magnitude of increased flux of the protein from synthetic sites to the extracellular space, described by a turnover index, S. The influence of sampling interval and analytical precision on the reliability of the estimated parameter values was investigated in the ideal situation by using computer simulation. It was found that informative estimates of the secretion rate of an acute phase protein could be obtained provided a fairly high analytical precision with a coefficient of variation less than or equal to 0.01 and using a sampling interval between 0.5--2 h.


The effect of N-(methoxy polyethylene glycol) rho-hydroxymercuribenzamide, a polymeric sulfhydryl group reagent, on spermatozoan motility.

The water-soluble, polymeric sulfhydryl group reagent N-(methoxypolyethylene glycol) rho-hydroxymercuribenzamide (Mw 5000) inhibited the motility of human spermatozoa. Its activity profile was very similar to that of rho-hydroxymercuribenzene sulfonate, a charged sulfhydryl group reagent that is a very poor membrane penetrant. These results suggest that functional sulfhydryl groups of the spermatozoan membrane are easily accessible and externally oriented on the membrane surface.


Effects of prazosin and phentolamine on cardiac presynaptic alpha-adrenoceptors in the cat, dog and rat.

In cats, dogs and rats, clonidine (20.0 microgram/Kg, i.v.) reduced the sustained tachycardia evoked by a continuous electrical stimulation of cardiac sympathetic nerve fibres. This effect of clonidine resulting from stimulation of cardiac presynaptic alpha-adrenoceptors could be completely antagonized by phentolamine and prazosin in the dog and the cat, whereas in the rat, prazosin, unlike phentolamine, failed to effectively inhibit the clonidine induced reduction in heart rate. The calculated doses of phentolamine and prazosin needed to produce a 50% antagonism were of similar magnitude in the dog, but in the cat prazosin was about three times less potent than phentolamine. As opposed to clonidine, LD 3098, a potent vascular postsynaptic alpha-adrenoceptor stimulating imidazoline, and phenylephrine (20.0 microgram/Kg, i.v.) did not significantly alter the sympathetic tachycardia in the pithed rat and the spinal dog. These results indicate that compounds stimulating postsynaptic alpha-adrenoceptors may be inactive as agonists of cardiac presynaptic alpha-adrenoceptors within the same animal species. Furthermore, a compound showing selective activity as an antagonist of vascular alpha-adrenoceptors in one species may not necessarily exert the same selective effects in other animals as demonstrated here for prazosin.


[Diseases caused by diisocyanates. 1. Irritation of the respiratory system and skin].

Toluylene diisocyanate (TDI) has an uncommon importance in the production of irritation of respiratory system. In the last few years less volatile isocyanate compounds have been substituted for TDI. Commercial available diphenylmethane diisocyanate (MDI) and polymethylene polyphenyl isocyanate (PAPI) were analyzed in 1971. The analysis indicated the presence of 21% TDI in several samples. It is evident, with respect to the relatively high vapor pressure of TDI, that such impurities will be able to cause air concentrations in excess of the current threshold limit value (TLV) of 0.02 ppm. Liberation of TDI by heat from varnish insulinations is a difficult problem. This problem was described as "an old hazard in new guise". Especially, in a soldering process on polyurethane coated wire, excess of TLV is possible in unfavourable conditions. By the regulation that the concentration of TLV must not exceed 0.02 ppm and by the use of new non-volatile isocyanate the risk of acute and subacute intoxications has considerably subsided. Chronic irritations of respiratory system and asthma-like diseases, however represent still an unsolved problem.


[Diseases caused by diisocyanates. 2. Diisocyanate asthma].

Diisocyanates are widely used as polymerizing agents in the production of polyurethanes. Exposure to high concentrations of toluene diisocyanate (TDI) causes the development of toxic-irritative symptoms in the respiratory tract. It has been observed that some workers, after repeated exposures and a latent period of months to years, develop asthmatic symptoms upon re-exposure to minute concentrations of diisocyanates. An immunological mechanism for this typical extrinsic asthma has not been adequately documented.


[Effects of morphine on the activity of various dorsal horn neurons of the spinal cord involved in nociception].

In spinal preparation, morphine exerts a specific direct inhibitory action on the activities of dorsal horn neurones induced by noxious stimuli. The effect of morphine is preferential for the responses evoked by A delta and C fibre afferents and its specificity of action has been demonstrated pharmacologically in terms of isomerism, dose dependency and reversal of the inhibitions by opiate antagonists. These results are in good agreement with recent data related to the localization at the spinal level of opiate receptors, and terminal rich in Enkephalin and substance P. Numerous behavioural and pharmacological investigations suggest that morphine is also acting at the level of the brainstem by reinforcing the activity of descending control systems which modulate the transmission of noxious inputs at the spinal level. However this second modality of action remains extremely difficult to demonstrate from an electrophysiological point of view.


Effects of cholera toxin on thyroid cyclic AMP-dependent protein kinase and ornithine decarboxylase activities.

Cholera toxin activated beef thyroid cyclic AMP-dependent protein kinase in a dose (0.2 to 8 microgram/ml)-related fashion. Thus, when beef thyroid slices were incubated with toxin (8 microgram/ml) for 90 minutes and then assayed for protein kinase, the activity ratio (i.e. -cyclic AMP/+cyclic AMP) increased from 0.32 +/- 0.02 to 0.77 +/- 0.06. The toxin (5 microgram/ml)-induced increase was abolished by inclusion of ganglioside GM1 in the incubation medium (I50, 0.7 microgram/ml), whereas, gangliosides GD1a and GT1 were without effect. In contrast, TSH-activated protein kinase was unaffected by ganglioside addition. Cholera toxin increased rat thyroid ornithine decarboxylase (ODC) activity in-vitro in a dose (0.1 to 10 microgram/ml)-related fashion [basal, 100 cf cholera toxin (10 microgram/ml), 1500 pmol 14CO2/g tissue/30 min]. The toxin (1 microgram/ml)- (but not TSH-) induced increase in ODC was abolished by inclusion of ganglioside Ga and GT1 were without effect. Cholera toxin stimulation of ODC was inhibited by indomethacin or iodide as are the stimulatory effects of TSH or dibutyryl cyclic AMP. These results demonstrate that although there are differences in the TSH and cholera toxin responses with respect to receptor (ganglioside) interaction, they nevertheless elicit similar intracellular responses in thyroid.


Subcellular localization of dopamine in the anterior pituitary gland of the rat: apparent association of dopamine with prolactin secretory granules.

The subcellular compartmentalization of endogenous dopamine in the anterior pituitary gland of the rat was investigated using continuous sucrose density gradient centrifugation. When anterior pituitary homogenates were layered on continuous sucrose density gradients (1.0--2.0 M) and centrifuged for 60 min at 40,000 X g, dopamine recovered from the gradients was associated with two sets of subcellular particles. The particles in one set were recovered near the top of the gradient, whereas those in the other set were recovered near the bottom of the gradient in the region where particles containing PRL were also found. In fact, these dense dopamine-containing particles could not be separated from those particles which contained PRL. These findings were suggestive that dopamine and PRL were present in the same particle, viz. the PRL secretory granule. This interpretation was further strengthened when it was established that the PRL-containing granules were separable on the gradient from granules which contained GH, LH, FSH, ACTH, and TSH. When [3H]dopamine was added to the solution in which the anterior lobes were homogenized, no radio-activity was found to be associated with the dense dopamine-containing particles. Also, the addition of a large excess of nonradiolabeled dopamine at the time of homogenization did not influence the amount of dopamine associated with the dense particles. Thus, the apparent association of dopamine with PRL secretory granules was not an artifact of the homogenization process per se. Therefore, it is concluded that an association exists between intracellular dopamine and the PRL secretory granule.


Plasma free tryptophan, brain serotonin, and an endocrine profile of the genetically obese hyperglycemic mouse at 4--5 months of age.

Genetically obese hyperglycemic mice (ob/ob) were compared with their nonlittermate lean controls at 4-5 months of age with regard to brain serotonin, pituitary ACTH content, and circulating levels of glucose, glucagon, insulin, TSH, T3, T4, total tryptophan and free tryptophan. Brain serotonin pituitary ACTH content, and plasma insulin, glucose, total tryptophan, and free tryptophan were all significantly higher in obese mice than in the controls. TSH, T3, and T4 were not significantly different in obese mice vs. controls, suggesting that the obese mouse is euthyroid. Fasting improved but failed to normalize the glucose and insulin levels or insulin to glucagon ratios. Since serotonin is an important neurotransmitter with regard to hypothalamic-pituitary function and since its levels in the brain are dependent on the availability of tryptophan, the findings of elevated levels of free tryptophan in the plasma and serotonin in the brain of the obese hyperglycemic mouse may help to explain some of the previously observed abnormalities of pituitary hormone secretion in these animals.


Sulfhydryl reagent-induced insulin release and 45Ca++ fluxes in Syrian hamster insulinoma cells.

Dispersed single cell suspensions of Syrian hamster insulinoma cells were used to study the effects of a variety of sulfhydryl-binding reagents on insulin release and 45Ca++ flux. Incubation of cells with several organic mercurials resulted in a rapid increase in 45Ca++ uptake as well as increased efflux in cells which had been prelabeled with 45Ca++. Concomitant with increased calcium uptake was a 4- to 5-fold increase in insulin released into the medium. Incubation with alkylating reagents such as iodoacetamide and N-ethyl maleimide or dithiol reagents such as 5,5'-dithiobis (2-nitrobenzoic acid) failed to stimulate either 45Ca++ flux or insulin release. Elimination of medium calscium or preincubation of cells with N-ethyl maleimide resulted in approximately 50% inhibition of mercurial-induced insulin release from these cells. 8-(N,N2-diethylamino)Octyl-3,4,5-trimethoxybenzoate or alpha-isopropyl-alpha [(N-methyl-N-homoveratryl)-gamma-aminopropyl]3,4,5'-trimethoxyphenylacetonitrite hydrochloride, agents which block potassium (40 mM)-stimulated calcium flux and insulin release, failed to inhibit mercurial-induced calcium flux or insulin secretion. These results indicate that sulfhydryl-binding reagents, through their interaction with critical thiol groups, promote insulin release in these insulinoma cells by inducing changes in calcium fluxes. It is possible that these thiol groups regulate calcium metabolism and, thus, insulin release under physiological conditions.


The role of sulfhydryl groups in thyrotropin binding and adenylate cyclase activities of thyroid plasma membranes.

The effects of sulfhydryl (SH) reagents on the binding of TSH to thyroid membranes and on membrane adenylate cyclase activity were studied. Diamide, a SH-oxidizing agent, enhanced [125I]iodo-TSH binding to thyroid membranes in a dose-dependent manner. On the contrary, dithiothreitol (DTT), a disulfide-reducing reagent, markedly inhibited this activity in a dose-dependent manner. These effects could not be attributed to a direct interaction between the reagents and the radioactive TSH. Scatchard plot analysis of the data was limited by curvilinear plots but suggested that diamide increases while DTT decreases the affinity of the receptors for TSH in thyroid membranes, neither reagent appeared to influence the capacity of TSH binding by membranes. The effects of diamide and DTT on adenylate cyclase activity of thyroid plasma membranes were just the opposite of their effects on TSH binding. Thus, diamide inhibited adenylate cyclase activity of thyroid plasma membranes while DTT increased basal enzyme activity and slightly but not significantly increased TSH-stimulated enzyme activity. The data suggest a possible physiological role for SH and disulfide groups of thyroid membranes in modulation of thyroidal activity.


Steroidogenesis in isolated cells and mitochondria of rat Snell adrenocortical carcinoma 494.

ACTH produced a 75% increase in pregnenolone biosynthesis from endogenous precursors in isolated cells prepared from the rat Snell adrenocortical carcinoma 494. On the addition of 24- and 25-hydroxycholesterol to the tumor cells, the rate of pregnenolone synthesis increased 10-fold but was insensitive to the presence of ACTH. Addition of lipoprotein cholesterol resulted in increased pregnenolone biosynthesis when ACTH was present. High density lipoprotein cholesterol appeared to be internalized and used for steroidogenesis preferentially to low density lipoprotein cholesterol. The cholesterol ester hydrolase activity of the cytosolic fraction of the tumor was found to be extremely low compared to that of the normal adrenal cell. These results, noting also the low cholesterol content of the tumor cells, suggested that the lack of availability of cholesterol was the factor responsible for the poor steroidogenic response of the cells to ACTH. The major steroid product of the tumor cells was determined to be deoxycorticosterone. This correlated with the low levels of steroid 11-beta-hydroxylase activity detected in the adrenal tumor mitochondria compared to the mitochondrial cholesterol desmolase activity. Little of the mitochondrial cytochrome P-450 appeared to function in a steroid 11-beta-hydroxylase complex.


Effects of insulin on glucose metabolism and glucose transport in fat cells of hormone-treated hypophysectomized rats: evidence that growth hormone restricts glucose transport.

In earlier studies we have shown that insulin does not stimulate glucose incorporation in adipocytes of hypophysectomized (hypox) rats. Basal glucose incorporation is decreased, although basal 3-O-methylglycose transport is very rapid and cannot be further stimulated by insulin. In this study we treated hypox rats with human GH, ACTH, and T3, alone or in combination, and examined the effects of insulin on glucose incorporation into fat cells and on 3-O-methylglucose transport. The results show that chronic administration of T3 alone to hypox rats partially restores glucose incorporation into fat cells and, in combination with ACTH, completely restores this incorporation. The two hormones have no effect on the glucose carrier system. The transport rate under T3 and ACTH replacement therapy continues to proceed at a maximal rate, so that basal glucose incorporation is high but not further enhanced by insulin. In contrast, administration of human GH to hypox rats does not influence glucose incorporation but has a marked effect on glucose transport. The basal glucose transport rate returns toward normal and again responds to insulin. This suggests 1) that enzyme activities responsible for the lipogenetic capacity of the fat cell are decreased in hypox rats and returned toward normal by the combined T3/ACTH treatment, and 2) that the limitation of glucose transport in the fat cell is controlled by GH. GH seems to induce a change of the glucose-carrier system; it leads to a restriction of glucose transport, which is acutely modulated by insulin.


Evaluation of histological classification in early gastric cancer (an analysis of 300 cases).

Using the Lauren approach of histological classification, 300 cases of early gastric cancer (EGC) were analysed with regard to their age distribution, to predisposing lesions, including gastritis, and survival rate. The average age of onset for EGC of the diffuse type is 56, ten years earlier than for the intestinal type. There was a significantly high percentage of EGC of the diffuse type without gastritis. On the other hand, gastritis in pernicious anaemia falls in the high-risk group. The survival rate in our cases is 98%, corrected for age. The results show that it is of utmost importance to differentiate between the histological types of gastric carcinoma; for there may be indeed a difference in pathogenesis and aetiology.


Investigations on the lung and kidney function in workers exposed to cadmium.

The kidney seems more sensitive to the chronic effect of cadmium than the lung. Only minor impairments of lung function (mild form of obstructive lung disease) were found after long-term occupational exposure (less than 20 yr) to moderate concentration of cadmium oxide dust and fume. This conclusion, cannot, however be extrapolated to acute or subacute inhalational exposure. The nephrotoxicity of cadmium consists in a tubular dysfunction characterized by an increased excretion of beta 2-microglobulin and giving rise to the classical tubular proteinuria and in a glomerular dysfunction evidenced by an increased excretion of high molecular weight proteins and increased levels of beta 2-microglobulin and creatinine in plasma and giving rise to a glomerular type proteinuria. These renal changes were mainly found in workers whose cadmium concentration at time of the survey exceeded 1 microgram Cd/100 ml in blood and 10 microgram Cd/g creatinine in urine. It should, however, be stressed that higher levels of Cd in blood and in urine are not necessarily associated with the presence of excessive proteinuria. In newly exposed workers, the Cd level in blood increases progressively to a plateau after several weeks. Cadmium level in urine fluctuates more. In workers exposed for several months to an airborne concentration exceeding 200 microgram/m3, Cd concentration in urine seems mainly influenced by recent exposure.


A study of the relationship between cadmium concentrations in urine and renal effects of cadmium.

The relationship between urinary cadmium concentration expressed as microgram/g creatinine and renal effects of cadmium exposure was studied in 542 inhabitants over 20 years of age who lived in the Jinzu River basin which is polluted by cadmium. Cadmium concentration in urine was employed as the index of cadmium exposure. Total protein with glucose, beta 2-microglobulin, retinol binding protein, and proline served as indices of renal effects. The prevalent rate of high beta 2-microglobulin excretion was the highest among these indices both in men and women. Prevalence rates of indices of renal effects increased proportionally with increasing cadmium concentrations in urine and probit linear regression lines could be calculated between them. The urinary cadmium concentrations corresponding to 1% prevalence rates of indices of the renal effects were calculated by the regression line. For beta 2-microglobulin, 3.2 microgram Cd/g creatinine and 5.2 microgram Cd/g creatinine were obtained in men and women, respectively. This method may be useful for evaluating the risk of renal damage of exposed inhabitants.


Exposure and accumulation of cadmium in populations from Japan, the United States, and Sweden.

Studies were carried out in Japan, United States, and Sweden regarding comparability of analytical methods for cadmium, daily intake of cadmium via food, daily amount of cadmium in feces, concentrations of cadmium in different tissues and the body burden of cadmium, urinary excretion of cadmium and cadmium concentrations in blood. It was found that the cadmium intake via food among adults is about 35 mug/day in Japan (Tokyo) and about 17 mug/day in the U.S. (Dallas) and Sweden (Stockholm). It varies with age in a way similar to calorie intake. Body burden increases rapidly with age. The half-time of cadmium is longer in muscles than in liver or kidneys. In the cross-sectional population samples studied (smokers and nonsmokers mixed) the average cadmium body burden at age 45 was about 21 mg in Japan, 9 mg in the U.S., and 6 mg in Sweden. Among nonsmokers in the U.S. and Sweden the body burden at age 45 was about 5-6 mg. The difference in average body burden for smokers and nonsmokers is explained by differences in smoking habits. Cadmium excretion in urine was closely correlated with body burden and about 0.005-0.01% of body burden is excreted daily in urine. Cadmium concentration in the blood was a good indicator of average recent intake over a 3-month period. Neither blood cadmium nor urine cadmium changed immediately after an increase of exposure level.


Recent results of health examinations on the general population in cadmium-polluted and control areas in Japan.

Health examinations by a revised method aimed at detecting renal tubular dysfunctions more effectively were conducted on the general population aged 50 years and over in cadmium-polluted areas (1826 persons) and control areas (1611 persons) in four prefectures in Japan in 1976. Although detailed analysis of the data is not yet completed, some of the results obtained are described here. The prevalence of glucosuria and low molecular weight proteinuria, frequency of decreased % TRP, and cadmium concentrations in urine are higher in the cadmium-polluted areas than in the controls. Clinically diagnosed cases with tubular dysfunctions seem to follow the same trend as above, though  these cases are very few in number and they are limited to the advanced age group. When cadmium concentration in rice is taken as an index, a dose-response relationship is not necessarity explicit in the observations by prefecture. However, suggestive data are obtained in the observation by district in one cadmium-polluted area.


Adrenal-testis interaction in the stallion.

The authors describe the short and long term effects of synthetic adrenocorticotrophin and of cortisol on peripheral plasma testosterone concentrations in 2 stallions. A single injection of either hormone temporarily raised plasma testosterone concentrations but repeated injection (twice daily for 5 days) depressed plasma testosterone concentrations. Cessation of treatment was followed by a rise in plasma testosterone to concentrations higher than those in the pretreatment period. These findings are briefly discussed.


The ATP synthetase of Escherichia coli K12: purification of the enzyme and reconstitution of energy-transducing activities.

The ATP synthetase of Escherichia coli K12 was purified by a simple procedure. The dicyclohexylcarbodiimide-sensitive ATPase activity was enriched 21-fold. The ATP synthetase preparation contained the eight polypeptides (alpha, beta, gamma, a,delta, b,espilon, c) of the enzyme and a residual contamination (4% of the total protein) as shown by dodecylsulfate/polyacrylamide electrophoresis. The polypeptide c was specifically labelled with [14C]dicyclohexylcarbodiimide. Energy-transducing activities were reconstituted from soybean phospholipids and the purified enzyme. The proteoliposomes exhibited a significantly higher ATP-32Pi exchange activity and a higher proton-translocating activity as compared to the untreated membranes.


Nucleotide sequence of the Hind-C fragment of simian virus 40 DNA. Comparison of the 5'-untranslated region of wild-type virus and of some deletion Mutants.

We report here the nucleotide sequence of the wild-type simian virus 40 (strain 776) restriction fragment Hind-C-P1 DNA and of the homologous region of various mutant DNAs which lack part of this fragment. During this work, we detected between EcoRII fragments N and G an additional, 17-base-pair EcoRII fragment, fragment P, which had previously been overlooked. Also, an additional dTpdG dinucleotide at residues L 339--340 was observed by sequence analysis of the DNA minus (E) strand; the presence of this dinucleotide was masked on sequencing patterns of the plus strand due to the persistence (during gel electrophoresis) of some secondary structures in the strand's 5'-terminal region. These nucleotide additions raise the total length of SV40 DNA to 5243 base pairs. The longest tandemly repeated segment in SV40 DNA now extends over 72 base pairs. SV40 deletion mutants dl 893 and dl 894 and SV40 strains Rh 911 and 1801 all lack an identical 72-base-pair-long DNA segment in the Hind-C region. This deletion corresponds precisely to one of the two aforementioned large tandemly repeated sequences. Mutant dl 895 lacks 66 base pairs, 63 of which are part of the former repetition. All these mutants, except dl 895, very probably were generated by an intramolecular, homologous recombination event. The 40-base-pair deletion in mutant dl 1811 includes the major capping site of SV40 late RNA. dl 1812 lacks only three base pairs, which are part of the overlapping HhaI and HpaII restriction sites at position 0.725--0.726.


Transcription by eukaryotic RNA polymerases A and B of chromatin assembled in vitro.

Chromatin was assembled in vitro from simian virus 40 DNA form I and the calf-thymus four histones H2A, H2B, H3 and H4. Transcription with calf thymus RNA polymerases A and B (I and II) was greatly inhibited. Nucleosomes were found to inhibit both RNA chain initiation and elongation. The inhibition of elongation could be relieved by increasing ionic strength, suggesting that electrostatic interactions between histone octamer and DNA have to be broken for RNA polymerase to transcribe DNA organised into nucleosomes.


Affinity chromatography of the beta-adrenergic receptor from turkey erythrocytes.

The beta 1-adrenergic receptors of turkey erythrocyte membranes have been identified by binding of the radioactively labeled antagonist (--)-[3H]dihydroalprenolol, solubilized by treatment of the membranes with the detergent digitonin, and purified by affinity chromatography. Binding of (--)-[3H]dihydroalprenolol to the membranes occurred to a single class of non-cooperative binding sites (0.2--0.3 pmol/mg protein) with a equilibrium dissociation constant (Kd) of 8 (+/- 2) nM. These sites were identified as the functional, adenylate-cyclase-linked beta 1-adrenergic receptors on the basis of: firstly, the fast association and dissociation binding kinetics at 30 degrees C; secondly, the stereospecific displacement of bound (--)-[3H]dihydroalprenolol by beta-adrenergic agonists and antagonists; and thirdly, the order of potencies for agonists to displace bound tracer (isoproterenol congruent to protokylol greater than norepinephrine congruent to epinephrine) similar to the one found for adenylate cyclase activation, and typical for beta 1-adrenergic receptors. Treatment of the membranes with the detergent digitonin solubilized 30% of the receptors in an active form. Digitonin solubilized also adenylate cyclase activity with a yield of 20 to 30%, provided the membranes were first treated with an effector known to produce a persistent active state of the enzyme: e.g. sodium fluoride. Binding sites for guanine nucleotides ([3H]p[NH]ppG) were solubilized as well. Their concentration (24 pmol/mg protein) was in large excess over the concentration of solubilized receptors (0.30--0.45 pmol/mg protein). Solubilized receptors were purified 500--2000-fold by affinity chromatography with a 25 to 35% yield, using an alprenolol-agarose affinity matrix. Affinity purified receptors were devoid of measurable adenylate cyclase activity and guanine nucleotide binding sites, thus showing that receptors and adenylate cyclase are distinct membrane constituents, and that guanine nucleotides apparently do not bind directly to the receptor molecules. Membrane-bound, solubilized and purified receptors were sensitive to inactivation by dithiothreitol, but not by N-ethylmaleimide, suggesting that receptors are at least partly constituted of protein molecules, with essential disulfide bonds.


The relipidation of delipidated Na,K-ATPase. An analysis of complex formation with dioleoylphosphatidylcholine and with dioleoylphosphatidylethanolamine.

Enzymatically inactive, delipidated Na,K-ATPase from dogfish rectal glands was titrated with dioleoylphosphatidylcholine and with dioleoylphosphatidylethanolamine. The process of relipidation has the following characteristic properties. Enzymatic activities reappear independently of each other: first the phosphatase, then the ATPase. The properties of the phosphatase regenerated depend on the ratio of lipid/protein used; the ATPase seems to be independent of this ratio. The simplest model that is consistent with the above results and with the shapes of the titration curves, has the following requirements. Firstly, the enzyme is composed of two subunits that, as far as lipid binding is concerned, are identical and independent of each other. Secondly, lipid adds onto the enzyme as preformed clumps of 25 molecules of phosphatidylcholine or 18 molecules of phosphatidylethanolamine. Thirdly, each subunit binds two clumps of lipid, and binding shows positive cooperativity. Fourthly, when either subunit becomes saturated with lipid, the enzyme exhibits one form of phosphatase. Fifthly, when both subunits are saturated with lipid, the enzyme exhibits a second form of phosphatase and ATPase. The data and their analysis according to this model lead to the suggestion that Na,K-ATPase is a functional dimer, the interaction between subunits being influenced by the Na+ and K+ concentrations in the medium: K+ favouring the functional independence of the subunits and Na+ favouring their functional interaction.


Isolation of mitochondrial succinate: ubiquinone reductase, cytochrome c reductase and cytochrome c oxidase from Neurospora crassa using nonionic detergent.

The electron transfer complexes, succinate: ubiquinone reductase, ubiquinone: cytochrome c reductase, and cytochrome c: O2 oxidase were isolated from the mitochondrial membranes of Neurospora crassa by the following steps. Modification of the contents of the complexes in mitochondria by growing cells on chloramphenicol; solubilisation of the complexes by Triton X-100; affinity chromatography on immobilized cytochrome c and ion exchange and gel chromatography. Ubiquinone reductase was obtained in a monomeric form (Mr approximately 130 000) consisting of a flavin subunit (Mr 72 000) an iron-sulfur subunit (Mr 28 000) and a cytochrome b subunit (Mr probably 14 000). Cytochrome c reductase was obtained in a dimeric form (Mr approximately 550 000), the monomeric unit comprising the cytochromes b (Mr each 30 000), a cytochrome c1 (Mr 31 000), the iron-sulfur subunit (Mr 25 000), and six subunits without known prosthetic groups (Mr 9000, 11 000, 14 000, 45 000, 45 000, and 52 000). Cytochrome c oxidase was also isolated in a dimeric form (Mr approximately 320 000) comprising two copies each of seven subunits (Mr 9000, 12 000, 14 000, 18 000, 21 000, 29 000, and 40 000). The complexes were essentially free of phospholipid. Each bound one micelle of Triton X-100 (Mr approximately 90 000). After isolation, the bound Triton X-100 could be replaced by other nonionic detergents such as: alkylphenyl polyoxyethylene ethers, alkyl polyoxyethylene ethers and acyl polyoxyethylene sorbitan esters.


Enzymology of ubiquinone-utilizing electron transfer complexes in nonionic detergent.

The enzymology of isolated succinate: ubiquinone reductase and ubiquinone: cytochrome c reductase in nonionic detergents (alkyl polyoxyethylene derivatives) was studied. In the membrane the two multiprotein complexes and their hydrophobic substrates ubiquinone and dihydroubiquinone, are embedded in a common lipid bilayer. In detergent solutions the complexes are each inserted into micelles. Detergent micelles also serve as a solvent for the complexes hydrophobic substrates. As a consequence the isolated complexes are in a discontinuous phase with respect to their hydrophobic substrates and with respect to each other. Three types of assays were used. Firstly, single enzyme assays in which the hydrophobic substrates had to transfer from free micelles to the complex-bound micelles in order for enzyme reactions to occur. Secondly, assays in which the enzymic reactions were coupled to auxiliary nonenzymic reactions which rapidly converted the hydrophobic products back into substrates within the complex-bound micelle. Dichloroindophenol was used for the oxidation of dihydroubiquinone and dihydroduroquinone for the reduction of ubiquinone. Thirdly, assays in which the succinate: ubiquinone reductase reaction was coupled with the ubiquinone: cytochrome c reductase reaction. With the first type of assay, the kinetics of the substrate transfer reaction was dependent upon the type of detergent. In detergents with small polyoxyethylene head groups the transfer reactions were rate-limiting, and in detergents with large polyoxyethylene head groups the transfer reactions were fast and the enzymic reactions were rate-limiting...


Membrane retrieval in epithelial cells of isolated thyroid follicles.

Follicles from rat and pig thyroid glands were isolated by digestion with collagenase. The epithelial cells of isolated follicles maintain their structural and functional polarity as shown by incorporation of 3H-leucine and autoradiography. To trace the fate of surface membrane, isolated follicles were opened, stimulated with thyrotropin and incubated for various time intervals with cationized ferritin (CF), uncharged dextran, native ferritin (NF), and latex spheres (0.5 mum in diameter) which were either pre-coated with CF or added together with CF. Uncharged dextran and native ferritin did not bind to the luminal cell membrane, were taken up in small amounts and accumulated in lysosomes; anionic NF was not found in Golgi cisternae in contrast to uncharged dextran which occassionally reached a few Golgi stacks. CF bound rapidly and in clusters to the luminal plasmalemma, preferentially to coated pits, was taken up by endocytosis, accumulated in lysosomes after 5 min and reached the Golgi cisternae after 30 min. Latex spheres were taken up by engulfment through fusion of microvilli and reached the lysosomes. CF particles coating the latex spheres may detach at this station and reach the Golgi cisternae. The findings show that the route of small tracers depends on the charge of the tracer, in agreement with results obtained by Farquhar [8]. Vesicles carrying NF can be traced to lysosomes only, whereas vesicles containing uncharged dextran or - more conspicuously -CF also fuse with Golgi membranes. Large tracers (latex beads) reach only the lysosomes, but CF taken up with them may move to Golgi cisternae.


Maintenance of Fc receptors in hybrid cell lines obtained by fusing mouse splenic macrophages with mouse myeloma cells.

Mouse splenic macrophages were fused with cells of the mouse myeloma line P3-X63-Ag8 in the presence of inactivated Sendai virus. Two continuously growing hybrid cell lines were established from fusion mixtures. These hybrid cell lines exhibited macrophage-like morphology and continued to express macrophage derived Fc receptor activity even after prolonged culture in vitro.


Studies on the central pressor activity of dibutyryl cyclic AMP.

The effects of the central administration (i.c.v.) of dibutyryl cyclic AMP (DBcAMP) on arterial blood pressure and heart rate were studied in the rat. The cyclic nucleotide produced a dose-dependent rise in blood pressure and an accompanying tachycardia. Maximal pressor effect of a single dose of DBcAMP (350 micrograms) was observed 25 min post injection and the duration of the response was 60 min. Chemical degeneration of central catecholaminergic neurons with 6-hydroxydopamine treatment abolished the pressor response to DBcAMP. Partial chemical degeneration of central serotonin pathways with 5,6-dihydroxytryptamine did not significantly alter the effect. The pressor activity of the cyclic nucleotide was attenuated by central administration of phentolamine. Spinal cord section at the C6-C7 level or peripheral chemical degeneration of catecholaminergic neurons combined with bilateral adrenalectomy abolished the pressor response to DBcAMP. It was concluded that the pressor response to i.c.v. administration of DBcAMP is mediated by central catecholaminergic pathways and is the result of an increase in efferent sympathetic outflow and release of catecholamines from the adrenal medulla.


Pharmacological activities of acetal derivatives of hemicholinium no. 3.

Acetal derivatives of hemicholinium no. 3 (HC-3) were synthesized and their chemical structures were confirmed by spectrophotometric evidence and elemental analysis. The acetals elicited a biphasic pattern of neuromuscular inhibition in the cat: (a) an immediate inhibition (early phase block) was judged to be curare-like as responses to acetylcholine (close i.a.) were abolished by acetal administration and reversal of inhibition was effected by neostigmine (25 micrograms/kg); (b) a slow, progressive inhibition of transmission (late phase block) was considered HC-3-like as it occurred only during high frequency stimulation and was antagonized by small doses of choline (1--3 mg/kg). In comparison to HC-3, significanlty greater curare-like activity was noted following acetal administration in vivo and in vitro. Mouse toxicity and cat nerve--muscle studies revealed the acetals to be HC-3-like but 1/2 to 1/3 as active as the parent compound. Cholinesterase inhibition by HC-3 and the acetals was low (I50 greater than 0.1 mM) and did not account for differences in activities. Acetal-elicited hypotension in the cat was attributed to postsynaptic ganglionic blockade and histamine release. Studies employing the 14C-(N-methyl) acetals furnished no evidence of the bioactivation (O-dealkylation) of the acetals to HC-3 in the cat in vivo or in cat liver in vitro. Chromatographic analysis afforded no evidence of molecular modifications of 14C-HC-3 or of the 14C-acetals in these systems.


Direct chronotropic and inotropic responses to guanosine and five different guanine nucleotides in isolated perfused dog atria.

The effects of guanosine and of five guanine nucleotides on sinus rate and contractile force were investigated using the isolated, blood-perfused dog atrium preparation. Guanosine consistently induced a positive chronotropic and inotropic effect which was not suppressed by treatment with a potent beta-adrenoceptor blocking agent, carteolol. GMP, GDP and CTP induced a biphasic chronotropic and inotropic effect. The potencies for inducing the positive effects of GMP, GDP and GTP were almost the same. However, their rank order of potency for inducing negative effects was GTP greater than or equal to GDP greater than GMP. The negative responses to GTP were not inhibited by atropine. Cyclic GMP and dibutyryl cyclic GMP produced only slight negative chronotropic and inotropic effects but not consistently even at larger doses. The positive chronotropic and inotropic responses to norepinephrine were not modified by treatment with dibutyryl cyclic GMP.


Presynaptic GABA autoreceptors on GABAergic nerve endings of the rat substantia nigra.

The release of 3H-GABA evoked by exposure to 30 mM potassium during 1 min was found to calcium independent in the rat occipital cortex and calcium dependent in the substantia nigra. Exposure to either muscimol 1 microM or GABA 1 microM inhibited the potassium-evoked release of 3H-GABA from the substantia nigra but not from the occipital cortex. The inhibitory effect of muscimol 1 microM on the potassium-evoked release of 3H-GABA from the substantia nigra was significantly antagonized by picrotoxin 10 microM. Exposure to either 10 or 100 microM pricrotoxin alone did not affect the potassium-evoked release of 3H-GABA. These results are compatible with the presence of a negative feedback mechanism in bagaergic nerve terminals of the rat substantia nigra which is mediated by presynaptic GABA autoreceptors. In addition, these results emphasize the importance of the calcium-dependent nature of the release of the neurotransmitter for demonstrating the modulation of transmitter release through presynaptic receptors.


Muscarinic receptor binding in the rat adrenal medulla.

Muscarinic receptor binding sites are present in rat adrenal gland andare associated primarily with the medulla. A Scatchard analysis using the ligand [3H]-quinuclidinyl benzylate (eH-QNB) revealed a KD of 0.064 nM and a Bmax of 66 fmol/mg prot. Denervation of the adrenal gland had no significant effect on 3H-QNG binding. The presence of muscarinic receptor binding sites in the adrenal medulla is consistent with reports that muscarinic receptors play a role in the release of adrenal catecholamines and modulation of cycle 3',5'-guanosine monophosphate.


The actions of enkephalins are not modified by the "kininase-II" inhibitor captopril.

The "kininase-II" (EC 3.4.15.1) inhibitor captopril in concentrations up to 10(-4) M did not affect the actions of enkephalins on the isolated electrically stimulated guinea-pig ileum. Captopril (100 mg/kg i.p.) in mice did not influence stimulation-produced analgesia and did not show analgesic properties. It is concluded that "kininase-II" is not involved in enkephalin catabolism, since the pharmacological actions of enkephalins were not enhanced after inhibition of the enzyme by captopril.


Binding to alpha-adrenergic receptors: differential pharmacological potencies and binding affinities of benzodioxanes.

We have compared the influence of a series of benzodioxane alpha-adrenergic antagonists on 3H-WB-4101 and 3H-clonidine binding to alpha-receptor sites in the brain and peripheral tissues with their pharmacological properties. The drug specificity of 3H-WB-4101 binding is quite similar in central and peripheral tissues. Pharmacological potencies of benzodioxanes at postsynaptic alpha-receptors in the rat vas deferens correlate with potencies at 3H-WB-4101 but not at 3H-clonidine binding sites. These findings suggest pharmacological effects of these drugs are mediated by "alpha-1 postsynaptic receptors" labeled by 3H-WB-4101. For several benzodioxanes absolute pharmacological potencies at postsynaptic alpha-receptors of the rat vas deferens are substantially less than their potencies at 3H-WB-4101 sites. The potencies of benzodioxane analogues at 3H-clinidine binding sites are similar to their pharmacological potencies at presynaptic autoreceptors in the rat vas deferens.


Differential effect of potassium on the action of digoxin and digoxigenin in guinea-pig heart.

The effect of potassium on the binding of digoxin or digoxigenin to isolated Na+, K+-ATPase was compared with that of potassium on the positive inotropic action of the agents in guinea-pig hearts. The binding of digoxigenin to the enzyme in vitro was reduced to a greater extent by potassium than was the binding of digotoxin. The digoxigenin-induced increase in the force of contraction of left atrial preparations estimated at steady state was reduced at higher potassium concentrations. Potassium had a lesser effect when digoxin was used as the inotropic agent. In contrast, potassium concentrations. Potassium had a lesser effect when digoxin was used as the ininotropic agent. In contrast, potassium reduced the rate of development and also the rate of loss of the positive inotropic action of digoxin observed with left atrial and Langendorff preparations, respectively, to a greater extent than those of digoxigenin. The loss of the positive inotropic effect was more rapid with digoxigenin than with digoxin at each KCl concentration. These data support the contention that the extent of the interaction of digitalis with Na+,K+-ATPase determines the degree of the positive inotropic effect.


Effects of ACTH4-10 on synaptic transmission in frog sympathetic ganglion.

The influence of ACTH4-10, a behaviourally active fragment of adrenocorticotropic hormone (ACTH) devoid of endocrine activity, on synaptic transmission in the paravertebral sympathetic ganglion of the frog was investigated. Postsynaptic potentials evoked by electrical stimulation of pregnanglionic nerves were recorded using a sucrose gap method. Fast excitatory postsynaptic potentials (EPSPs), which are mediated via nicotinic cholinergic synapses, were not affected by 10(-6) M ACTH4-10. Application of ACTH4-10 in  a concentration as low as 10(-8) M for 60 min caused a marked augmentation of the amplitude of slow inhibitory postsynaptic potentials (IPSPs) which are mediated via dopaminergic synapses. The increase in amplitude developed gradually after a latency of 60--90 min and outlasted the application of the peptide. In addition, ACTH4-10 at 10(-6) M increased the hyperpolarising response of the ganglion to exogenous dopamine, as studied by a micro-application method. There was no significant effect of ACTH4-10 on the muscarinic cholinergic depolarising response of the ganglion towards exogenous acetylcholine. The behaviourally active vasopressin fragment DG-LVP (10(-6) M) had no effect on slow IPSPs. The results demonstrate that ACTH4-10 specifically affects slow synaptic inhibition in frog sympathetic ganglion, probably by acting upon the postsynaptic membrane. The possibility is discussed that ACTH4-10 affects one of the intermediate steps between dopaminergic receptor interaction and generation of the slow IPSP.


The effects of chemical sympathectomy with 6-hydroxydopamine on alpha-adrenoceptor and muscarinic cholinoceptor binding in rat heart ventricle.

The specific binding of 3H-dihydroergocryptine to alpha-adrenoceptors in membranes from the heart ventricle of rats chemically denervated with 6-hydroxydopamine was significantly decreased in comparison with control animals. In contrast the muscarinic cholinoceptor binding of 3H-quinuclinidyl benzilate was unchanged after chemical sympathectomy. These results are interpreted as indicating that the muscarinic receptors are principally postsynaptic but that a significant proportion of the alpha-adrenoceptors are located on noradrenergic nerve terminals.


Friend virus production and heme synthesis in primary mouse spleen cell cultures.

A cell culture method has been developed in which spleen cells from Friend virus (FV) infected mice can be studied for virus production as well as erythroid differentiation. Primary spleen cell cultures from plethoric Balb/c mice were initiated at 24, 48 or 73 h after FV infection. These cells manifested a well-defined wave of heme synthesis at approximately 64, 48, or 23 h, respectively, of cell culture. Assays for spleen focus-forming virus (SFFV) and helper murine leukemia virus (MuLV-F) production in these cultures revealed that the peak rates of production of both viruses occurred at essentially the same time as the peaks of heme synthesis. The time at which the peaks of virus production and heme synthesis occurred in vitro was related to the time interval after infection (80-105 h) rather than the time at which the cells were placed in cell culture or the number of hours of cell culture. Medium change experiments suggested that the temporal relation between heme synthesis and virus production was an intrinsic feature of FVP infected cells in this in vitro system.


Changes in oxygen radical scavenging by human blood cell lysates concurrent with viral infections.

The higher superoxide dismutase (SOD) levels found in human blood cells when Nitro Blue Tetrazolium (NBT) rather than Cytochrome C was used as the colorimetric detector of superoxide (O-2) was investigated. the NBT was found to react with oxygen radicals other than O-2, thus providing measurement of total oxygen radical scavenging ability. Also investigated was the biomodal distribution of SOD activity in lymphocytes and granulocytes from a randomly selected human populace. Human lymphocytes rapidly increased their SOD activity by two- to four-fold during respiratory viral distress. The response to certain infections was probably responsible for the bimodal distribution of SOD activity in the general populace. It was concluded that the chemical events that prepare lymphocytes and granulocytes for their role in defense against infections begin long before these cells are sequestered from circulation by the infections.


Mechanisms of compensation for vestibular deficits in the frog. I. Modification of the excitatory commissural system.

In hemilabyrinthectomized frogs excitatory responses of central vestibular neurons to electrical stimulation of the remaining vestibular nerve were recorded extra- and intracellulary at different stages (0, 3, and 60 days) after the operation. The output pattern of ipsilateral vestibular neurons sending an axon across the midline via the vestibular commissure to the deafferented nucleus did not change postoperatively. The synaptic efficacy of these commissural axons ending on partially deafferented vestibular neurons on the lesioned side increased with time. This enhanced synaptic potency was associated with a shortening in time to peak and duration and an increase in amplitude of the evoked EPSPs. As a result most vestibular neurons were readily excited by single shock stimulation of the contralateral vestibular nerve, a finding which was rarely observed in control animals. These plastic changes are explained by the assumption of reactive synaptogenesis. The consequences of this modification for the readjustment of static and dynamic vestibular reflexes are discussed.


Studies on the late-pre-beta-lipoprotein of human serum.

Very low density lipoprotein (VLDL) isolated from 3 healthy normolipidaemic subjects had a raised VLDL cholesterol to triglyceride ratio. The VLDL fractions gave 2 pre-beta-bands on agarose gel electrophoresis. Family study of the subjects appears to indicate sex linkage of this trait and a possible polygenic type of inheritance.


Mechanisms of heterotypic immunity against canine distemper.

Hep-2 cells infected with measles virus (MV) for as short as 6 h became refractory to superinfection with canine distemper virus (CDV) but not to vesicular stomatitis virus (VSV). The exact mechanism of such interference is unknown but probably occurs after virus attachment and penetration. These results verify the suggestion that virus interference may be a mechanism of heterotypic protection against canine distemper.


Inhibition of hepatic Na +, K + -adenosinetriphosphatase in taurolithocholate-induced cholestasis in the rat.

Na +, K + -adenosinetriphosphatase (Na +, K + -ATPase) activity was decreased in liver plasma membranes from rats in which cholestasis had been induced by i.v. administration of sodium taurolithocholate (5 mumoles/100 g b. wt). Incubation of liver plasma membranes with taurolithocholate (10--1300 muM) caused significant and dose dependent reductions of Na +, K + -ATPase activity at taurolithocholate concentrations above 100 muM. These findings lend support to the hypothesis that cholestasis induced by monohydroxy bile acids is at least partially the result of an inhibition of hepatic Na +, K + -ATPase activity.


The effect of halothane on the action of alcuronium in the dog.

The effect of halothane on the action of alcuronium on neuromuscular transmission was studied in the intact dog. Electrical and mechanical evoked responses from stimulation of the ulnar nerve were recorded. The administration of 0.5% halothane prolonged the duration of action of alcuronium by about 50%.


ACTH-induced hyperalgesia in rats.

The injection of ACTH 1--24 into the cerebral ventricles in rats markedly reduces the reaction time in the hot-plate test and the nociception threshold in the tail-stimulation test. Morphine antagonizes and naloxone potentiates this hyperalgesic effect of ACTH. It is proposed that ACTH peptides play a physiological role in nociception.


The inhibitory effect of paraquat on histamine and isoproterenol induced changes of cyclic nucleotides in rat lung slices.

The incubation of rat lung slices with paraquat ion (10(-4) M) had no effect on cAMP and cGMP levels of the rat lung slices. The preincubation with the same concentration of paraquat inhibited the cAMP elevating effect of histamine (10(-5) M) and isoproterenol (10(-5) M) and reduced the cGMP level to approximately 50% of the level obtained without preincubation with paraquat.


[Tercuronium, a new nondepolarizing myorelaxant with high activity and selective action].

Tercuronium is p',p"-bis-triethylammonium-p-terphenyl dibenzosulfonate. As a curarelike agent tercuronium is 4--8 times as potent as (+)-tubocurarine. The time of the development and lasting of the blocking effect of tercuronium is approximately the same as that of (+)-tubocurarine. In a blocking dose tercuronium does not exert any effect either on the vegetative ganglia or arterial blood pressure. Partial blocking of the transmission through the vegetative ganglia as well as an insignificant and short-term drop of arterial blood pressure are recorded after intravenous injection of 10 myoparalytic doses of tercuronium. The antagonism of neostigmine against tercuronium was more pronounced than against (+)-tubocurarine, pancuronium and gallamine.


[Nature of the hyperpotassic composition of the medium surrounding individual tissue mechanoreceptors].

The ouabain effect on potassium concentration in the Pacini corpuscle fluid was studied using continuous perfusion of cat's mesenteric receptors with ouabain in Ringer's bicarbonate solution. To check the solution penetration into the corpuscles, Na36Cl and (G-3H)-ouabain were used. 2 . 10(-2) mM ouabain reduced (K+) in corpuscle fluid. The high potassium concentration in Pacini corpuscle fluid seems to be due to Na+--K+-ATPase system.


[Adrenergic mechanisms of changes in the resistance and volume vessels of the small intestine in the presence of a reduced volume of circulating blood].

In humorally isolated small intestine, the 10--20%--decrease in the perfusing blood volume against the background of augmented vascular resistance of the small intestine, induces different shifts of the latter's vascular capacitance. At the blockade of small intestine's beta--adrenoreceptors the decrease does not practically tell on the character of shifts in the intestine's resistance and capacitance while the blockade of alpha--adrenoreceptors abolishes the augmentation of these parameters.


[Adrenergic regulation of thermogenesis in Siberian lemmings].

Siberian lemmings seem to have lesser noradrenaline (NA) calorigenic action and higher beta-adrenergic "asymmetry" of catecholamine calorigenic effects than the cold- and warmth-adapted laboratory rodents. Selective inhibition of catecholamine effects by beta-adrenoblocking drug propranolol, obvious potentiation of NA effect by caffeine, and compensatory increasing of thermoregulatory musclar contractile activity during the blockade of betaadrenoreceptors under cooling suggest the domineering of betaadrenergic mechanisms of thermogenesis in siberian lemmings and their high thermoregulatory lability.


[The characterization of nuclear triiodo-L-thyronine receptors of rabbit liver (author's transl)].

The binding characteristics and physical properties of the nuclear receptors of rabbit liver were studied. The receptors were extracted with 0.4M KCl from purified hepatic nuclei and were incubated with increasing doses of 125I-triiodo-L-thyronine (L-T3) and stable L-T3 for 2 hours at 20 degrees C. Scatchard analyses indicated that the association constant (Ka) of the nuclear receptors was 1.5 X 10(10)M-1, and that maximal binding capacity was about 0.12 pmol T3 per 1 ml of nuclear extract, equivalent to 1 g of liver or 0.2 pmol T3 per mg of protein of the nuclear extracts. The nuclear binding sites were specific for L-T3. When compared by the molar concentrations of hormone analogues required to produce 50% inhibition of L-T3 binding, the relative binding affinities of triiodothyroacetic acid (Triac), D-T3, L-T4 and D-T4 were, respectively, 1/2, 1/4, 1/50 and 1/170 of L-T3. The binding affinity of Triac for the isolated hepatic nuclei was also nearly half that of L-T3. Furthermore, to displace radioactive Triac from the binding sites, Triac was half as effective as L-T3. The molecular weight of the nuclear receptors was estimated to be about 40000 to 45000 by the elution profiles from a Sephadex G-100 column, and the sedimentation coefficient was slightly less than 4S on sucrose density gradient centrifugations. Both elution profiles from DEAE- and ECTEOLA- cellulose columns with a linear KCl gradient showed a sharp, narrow peak of radioactive T3 at about 0.15 to 0.2M KCl. The results obtained with rabbit liver were similar to those reported previously in rat liver except for the relative binding affinity for Triac. It is possible that this discrepancy may be due to a species-related difference.


[Increased liver nuclear triiodothyronine-receptors associated with mitochondrial alpha-glycerophosphate dehydrogenase activity in hyperthyroid rats (author's transl)].

The effects of triiodothyronine (T3) on binding characteristics of nuclear T3-receptors were examined in rats. The T3-receptors were extracted by 0.4M KCl from the liver nuclei isolated from the following 3 groups of rats: (1) thyroidectomized, (2) thyroidectomized and treated with 230 ng T3/100 g body weight/day for 3 days, and (3) thyroidectomized and treated with 40 micrograms T3/100 g body weight/day for 3 days. Their association constant (Ka) and maximal binding capacity (Cmax) for T3 were determined by Scatchard analyses with and without correction for endogenous T3. The amount of endogenous T3 bound to the nuclear extracts was estimated on the basis of a specific activity of [125I]-T3 injected 2 hours before sacrifice. It was demonstrated that Cmax values corrected for the endogenous T3 were 2.5 times greater in severely hyperthyroid than in hypothyroid rats. By contrast, corrected values for Ka remained unchanged in all 3 groups of rats, although uncorrected values were apparently decreased in the severely hyperthyroid rats. Validity of the correction was supported by the in vitro experiments preincubated with stable T3. The yields of nuclear protein-125I-T3 complex in the procedure of extraction and the contents of DNA per g of liver were nearly the same in the 3 groups. The increase in Cmax of the nuclear receptors was directly related to mitochondrial alpha-glycerophosphate dehydrogenase activity. The results obtained indicate that the Cmax of nuclear T3-receptors is increased in a severely hyperthyroid state, related to hormonal action, and that the correction for endogenous T3 is essential for an accurate estimation of Ka and Cmax.


[Sequential changes in the nuclear triiodothyronine receptors and mitochondrial alpha-glycerophosphate dehydrogenase activity after the administration of triiodothyronine (author's transl)].

The dynamics of the induction of nuclear triiodothyronine (T3)-receptors and mitochondrial alpha-glycerophosphate dehydrogenase (alpha-GPD) were studied in rat liver after a single injection of a large amount of T3. The maximal binding capacity (Cmax) and association constant (Ka) of the nuclear receptors were determined by Scatchard analyses with and without correction for endogenous T3 measured by radioimmunoassay. It was demonstrated that the administration of T3 induced sequential increases in the number of nuclear T3-receptors and alpha-GPD activity in the liver. The nuclear receptors were rapidly increased to 2.5 times the hypothyroid level and thereafter decreased with a half-life of about 2 days. A parallel change in alpha-GPD activity was noted after a lag period. The total amount of extracted nuclear proteins in the liver was increased 3 days after the administration of T3. It seems likely, therefore, that the T3-induced increase in nuclear receptors is responsible, at least in part, for the induction of this enzyme. The possibility is also suggested that the nuclear receptors may be a nonhistone protein selectively synthesized in an early stage of the hormonal stimulation. Following the injection of T3, marked changes in apparent Ka were seen when no correction was made for the amount of endogenous T3 bound to the extracted nuclear proteins. With the correction, however, Ka remained the same despite the changes in the nuclear receptors and enzyme activity. The results obtained provide further evidence for the hormonal modulation of the nuclear receptors which is closely linked with the hormonal effect.


[In vivo and in vitro studies on 18-hydroxy-11-deoxycorticosterone and 18-hydroxycorticosterone in normal subjects and in those with various adrenocortical disorders (author's transl)].

Simultaneous measurement of 18-hydroxy-11-deoxycorticosterone (18-OH-DOC) and 18-hydroxycorticosterone (18-OH-B) in the peripheral plasma was carried out on normal subjects and in patients with adrenocortical disorders. The mean plasma levels of 18-OH-DOC at 0800h in normal males and in the follicular and luteal phases of normal females were 8.2 +/- 3.9 ng/100 ml (Mean +/- SD), 7.8 +/- 2.6 ng/100ml and 11.5 +/- 2.8 ng/100ml, respectively. The corresponding levels of 18-OH-B in normal males and in the follicular and luteal phases of normal females were 10.3 +/- 4.2 ng/100ml, 12.4 +/- 4.5 ng/100ml and 13.8 +/- 4.1 ng/100ml, respectively. No differences between the sexes nor the phases of the menstrual cycle were confirmed. ACTH stimulation increased plasma concentrations of 18-OH-DOC and 18-OH-B by 5.1 and 4.4 times respectively, while dexamethasone markedly decreased these 2 steroids. An upright posture increased these steroids significantly. In patients with Cushing syndrome, plasma levels of these 2 steroids were rarely high in cases with adrenocortical hyperplasia and adrenocortical carcinoma, while they were usually within the normal range in adrenocortical adenoma. These 2 steroid levels were increased in primary aldosteronism, idiopathic hyperaldosteronism and congenital 17 alpha-hydroxylase deficiency. They were decreased in Addison's disease and the salt-loosing type of congenital 21 alpha-hydroxylase deficiency. Patients with congenital 21 alpha-hydroxylase deficiency (simple form) showed elevated levels of 18-OH-DOC and normal levels of 18-OH-B. In vitro production of 18-OH-DOC and 18-OH-B was studied by tissue slices of the normal adrenal cortex, adrenocortical carcinoma causing Cushing syndrome, aldosteronoma and nodular hyperplasia with hyperaldosteronism. In the normal adrenal cortex, the mean production rates of 18-OH-DOC and 18-OH-B were 31 and 26 ng/g tissue/hr, respectively. ACTH and angiotensin II significantly increased the production of both 18-OH-DOC and 18-OH-B. In adrenocortical carcinoma, the production of these steroids was markedly diminished and not stimulated with either ACTH or angiotensin II. Aldosteronoma tissue produced these 2 steroids 20 to 40 times that of the normal adrenal tissue and was significantly increased with the addition of ACTH and angiotensin II. Nodular hyperplasia with hyperaldosteronism produced much 18-OH-DOC and 18-OH-B, but did not respond to ACTH and angiotensin II.


Early effects of thyroidectomy and triiodothyronine administration on rat-liver mitochondria.

This work was undertaken to study the action exerted by thyroid hormones on mitochondria. By day 6 after thyroidectomy, the respective activities of two inner-membrane enzymes--succinate and beta-hydroxybutyrate cytochrome c reductases--had already dropped by 32 and 50%, whereas, in the outer membrane, the activity of rotenone-insensitive NADH-cytochrome c reductase did not change significantly. The decrease in the activity of the inner-membrane enzymes closely followed the disappearance of T3 and T4 from serum. 10 h after administration of 25 micrograms/100 g T3 to thyroidectomized rats, the activity of succinate and beta-hydroxybutyrate cytochrome c reductases and the oxygen consumption rate with succinate or beta-hydroxybutyrate were significantly increased, while, in the outer membrane, the activity of monoamine oxidase and rotenone-insensitive NADH-cytochrome c reductase remained unchanged. In the thyroidectomized rat, L-[3H]leucine incorporation in vivo is diminished in all the liver mitochondrial proteins, and especially in two constituents of MW 19 000 and 28 000. The radioactivity of these two components is also decreased in the normal rat treated with chloramphenicol, a specific inhibitor of mitochondrial protein synthesis. L-[14C]leucine incorporation in isolated liver mitochondria was significantly increased in the thyroidectomized rat, 10 h after T3 treatment. Thus, thyroid hormones have an early and preferential action on the mitochondrial protein synthesizing system and on the inner-membrane enzyme activities.


Evidence for glucocorticoid transport into AtT-20 cells. Selective effect of a sulfhydryl inhibitor on uptake.

The purpose of these experiments was to determine if membrane-mediated glucocorticoid uptake by the AtT-20 cell was sensitive to the sulfhydryl group inhibitor, p-chloromercuriphenylsulfonate (PCMPS). This agent was chosen because of its reported limited entry into the cell interior. Our experimentes showed that cells could be incubated with 1 mM PCMPS for 1 h before intracellular concentrations of the inhibitor were sufficient to affect the intracellular cytosol receptor. In contrast less than 15 min of treatment inhibited intact cell steroid uptake. The inhibition was completely reversed by dithiothreitol. These studies show inhibition of glucocorticoid uptake under conditions which do not affect the intracellular receptor and infer that the AtT-20 cell membrane-mediated uptake mechanism for glucocorticoids contains sensitive sulfhydryl groups.


Effects of cannabinoids on testosterone and protein synthesis in rat testis Leydig cells in vitro.

Various water-insoluble cannabinoids as well as SP-111A, the water-soluble derivative of delta 9-tetrahydrocannabinol (delta 9-THC), reduced hCG and dibutyryl-cAMP stimulated testosterone production by rat testicular Leydig cell preparations. With 0.15 microM (0.05 micrograms/ml) 8-beta-OH-delta 9-THC the inhibition was about 50% of stimulated testosterone synthesis. Dose-related inhibitions were apparent with other cannabinoids and their order of potency in inhibiting stimulated steroidogenesis by the interstitial cells in vitro was found to be: 8-beta-OH-delta 9-THC greater than or equal to 11-OH-delta 9-THC greater than CBN = CBD = CBG greater than or equal delta 9-THC = delta 8-THC. The non-stimulated, basal, steroidogenesis was not affected even with 15 microM cannabinoids. The incorporation of L-[U-14C]leucine into the protein of Leydig cells was markedly reduced by 15 microM cannabinoids under both basal and stimulated conditions. The inhibition of steroidogenesis as well as protein synthesis in rat testicular Leydig cell preparations by various cannabinoids cannot be correlated with their psychoactivity. The present data suggest that cannabinoids at very low concentrations may interfere directly in Leydig cells with both protein and testosterone synthesis, and thus with their function.


Substance P stimulation of amylase release by isolated parotid cells and inhibition of substance P induction of salivation by vasoactive peptides.

Substance P stimulation of salivation in rats has been studied as has its in vitro enhancement of amylase release by isolated parotid cells. The extent of the stimulation on amylase release by isolated parotid cells was dependent upon the concentration of substance P, with the minimum effective concentration being 1 nM. The substance P effect was detectable within 1 min after incubation and lasted for at least 50 min. Substance P stimulation was demonstrable at 25--37 degrees C but not at 0 degrees C. Adrenocorticotropic hormone (ACTH), thyrotropin-releasing hormone (TRH), vasopressin and neurotensin had no effect on amylase release. These results suggest that substance P may act directly on the parotid cells. Examination of the salivary-stimulating activity of fragments of substance P showed that the C-terminal octapeptide and (pyroglutamyl)hexapeptide were active, although less potent than substance P, whereas its free acid, C-terminal tetra- and tri-peptides were inactive. Vasopressin, angiotensin II and neurotensin could inhibit substance P induced salivation, whereas TRH, ACTH and somatostatin had no effect. Amylase activity per unit volume of saliva was not changed by the injection of vasopressin, angiotensin II or neurotensin. These vasoactive peptides did not affect substance P stimulation of amylase release by isolated parotid cells. The results indicate that vasopressin, angiotensin II and neurotensin inhibit the action of substance P on salivation at sites other than the parotid cells.


Insulin inhibition of lipolysis of human adipocytes: the role of cyclic adenosine monophosphate.

To gain information on the manner in which insulin suppresses lipolysis in man, isolated adipocytes, prepared from subcutaneous adipose tissue, were incubated with insulin (100 microunits/ml) alone and in combination with isoproterenol (10(-7) M or 10(-8) M). Cyclic AMP concentration was measured at 60 min; glycerol release, used as an index of lipolysis, was determined at 45 and 75 min. Insulin consistently reduced both basal and stimulated cyclic AMP and glycerol release: the degree of suppression of each was comparable. In subsequent experiments, the ability of insulin to suppress glycerol release stimulated by isoproterenol, theophylline, and dibutyryl cyclic AMP (dbcAMP), respectively, was compared. Insulin substantially reduced the raised levels of cyclic AMP and glycerol release prompted by isoproterenol and theophylline, but it had little effect on increases caused by dbcAMP. These findings support the view that reduction in cyclic AMP is an important component in the regulation of fat mobilization by insulin.


Hepatic adenoma associated with portasystemic shunting in a young woman.

The development of a hepatic adenoma is described in a young female of reproductive age who had documented portasystemic shunting and hyperestrogenemia. The intrinsic hyperestrogenemia and the increased hepatic arterial blood flow, both of which occurred as a result of the portasystemic shunting secondary to portal venous obstruction, may have been important factors in the development of her hepatic tumor. We would postulate that prolonged endogenous hyperestrogenemia, like exogenous sex steroids, may predispose to hepatic adenoma formation.


Intestinal absorption of cholecalciferol in alcoholic liver disease and primary biliary cirrhosis.

The intestinal absorption of (3H)cholecalciferol was studied in five patients with alcoholic liver disease, six patients with primary biliary cirrhosis, and 15 healthy subjects. The rate of appearance in plasma of (3H)cholecalciferol after oral ingestion and the subsequent appearance of (3H) polar metabolites in the alcoholic subjects were similar to those in the healthy subjects. In subjects with primary biliary cirrhosis the rate of appearance in plasma of (3H)cholecalciferol was significantly reduced. The rate of appearance of labelled polar metabolites of cholecalciferol was also lower in this group, suggesting that increased removal of labelled vitamin by conversion into more polar metabolites could not account for the reduced plasma (3H)cholecalciferol response. It is suggested that intestinal absorption of cholecalciferol is usually normal in alcoholic liver disease but impaired in primary biliary cirrhosis. Hepatic 25-hydroxylation is normal in alcoholic liver disease but may be defective in primary biliary cirrhosis.


Rôle of calcitonin in diarrhoea associated with medullary carcinoma of the thyroid.

Severe watery diarrhoea in a patient with medullary thryoid carcinoma and high calcitonin levels was associated with increased distal ileal flow. Plasma from the patient inhibited small intestinal fluid absorption in dogs without affecting mucosal cyclic AMP levels. When calcitonin was removed from the plasma, the effect in experimental bowel loops was abolished but could be restored on restitution of comparable hormone levels with pure calcitonin. These studies, combined with the failure to find evidence of increased prostaglandin activity in the patient, suggest that circulating calcitonin may be a major cause of the diarrhoea in medullary thyroid carcinoma.


Acute metabolic and circulatory effects of cigarette smoking in late pregnancy.

The acute metabolic and circulatory effects of cigarette smoking (two cigarettes of a standard brand) in the last trimester of pregnancy were studied. 12 subjects, all of whom where hospitalized because of pregnancy complications, volunteered to participate in the investigation. A statistically significant immediate increment in the maternal heart rate, systolic and diastolic blood pressure, fetal heart rate, plasma glucose and plasma cyclic AMP was noted. Plasma glycerol, plasma insulin and blood lactate did not change significantly. 30 min after smoking there was a small but significant increase in plasma nonesterified fatty acids, plasma beta-hydroxybutyrate and plasma C-peptide.


Paraprofessional role in a mental health continuing care program.

The authors report on a unique program in which paraprofessionals provide continuing care services to patients in a large public psychiatric hospital, both before and after discharge. Data from one hundred patients indicate that paraprofessional caseworkers effectively managed 85 percent of the patients' post-discharge problems without needing to directly involve their professional teammates.


Determination of the activity of nonspecific monophosphoesterases and nonspecific esterase in the liver of dogs in extrahepatic cholestasis treated surgically and with phenobarbital.

Histochemical researches on livers of 19 dogs were carried out. The dogs suffered from extrahepatic cholestasis which lasted for 28 days and was surgically induced. The dogs were divided into three groups, out of which two were given phenobarbital for 21 or 28 days. In all groups biliary obstruction was removed by surgical operation. It was shown that a combined surgical treatment with the application of phenobarbital has given better results than a surgical treatment alone.


Liver cell injury (bodies similar to Lafora's) in alcoholics treated with disulfiram (Antabuse).

Inclusions, structurally similar to Lafora's bodies, are described in the liver cells of three chronic alcoholic patients who stopped drinking after disulfiram treatment. The inclusions were strongly positive with PAS and methenamine silver stains. Shikata stain for HBsAg was negative. On electron microscopy the inclusions are not membrane-bound and contain glycogen beta-granules, secondary lysosomes containing lamellar structures, lipid droplets and filaments; the SER was almost completely lost. In the patient least affected, the cells bearing inclusions were predominantly periportal in location, as is usual in Lafora's disease. In the two other patients the change involved the whole lobule. The possibility of an induced carbohydrate metabolic disorder, which could be due to the disulfiram, a drug that interferes with the activity of several hepatic enzymes is discussed. The presence of appearances suggestive of SER breakdown could also be interpreted as a manifestation of 'disuse atrophy' due to alcohol withdrawal.


On the genetics of the human serum paraoxonase (EC 3.1.1.2).

Incubation of the sera of 799 nonrelated persons with paraoxon led to varying degrees of inhibition of the serum cholinesterase (EC 3.1.1.8) with residual activity between 0% and 67.4% of the initial activity. This is the result of a differing paraoxonase (EC 3.1.1.2) activity. The residual activities show a trimodal distribution. The results of studies of 99 families with children show that an autosomal dominant herdity factor is most likely. Consideration of the constellations of the activity values within the families can thus yield a stochastic external criterion. This, together with the shape of the distribution of the individual values, gives good statistical estimates for the distributions and frequencies of the three groups obtained by an iteration technique. Tests of association that take account of group membership show that residual activity does not depend on the blood groups A, B, O, and Rh, or on age. A conclusive argument for our assumption of three activity groups is that the resulting group frequencies are consistent with the Hardy-Weinberg rule.


Adherent cells as targets in mitogenic T-lymphocyte stimulation: replacement of lymphocyte-bound mitogens by nonmitogenic substances.

The sequence of events in mitogenic T-lymphocyte activation was investigated by employing purified mouse peritoneal adherent cells and T-lymphocyte populations. After treatment of adherent cells with the oxidizing agents sodium periodate or neuroaminidase plus galactose oxidase and removal of the mitogens, the cells acquired the ability to stimulate purified T-lymphocytes added subsequently. For this process of T-lymphocyte activation, the additional presence of the membrane interacting substances polyethylene glycol, dextran, nonmitogenic Helix pomatia agglutinin, or inactivated sendai virus was required. Lysates of mitogen-treated peritoneal adherent cells were able to replace the adherent cells in the cultures.


Altered immune reactivity in encephalomyocarditis-M variant (EMC-M)-induced diabetes in mice.

Approximately 50% of DBA/2j male mice infected with EMC-M develop glycosuria with suppressed immune responsiveness to the mitogens PHA, Con A and PWM. The greatest degree of suppression was noted with Con A treated cultures. In vivo treatment with Levamisole, a known T cell stimulant, especially increased the response to Con A but exacerbated the diabetogenic state as reflected by increased occurrence of glycosuria. Virus infected mice, given anti-lymphocyte serum (ALS), showed marked amelioration of the diabetic syndrome. Indomethacin, a non-steroidal anti-inflammatory agent as well as a prostaglandin-synthetase inhibitor, partially reversed the in vitro splenic cell suppression of infected animals.


Effect of silica on the innate resistance of inbred mice to Salmonella typhimurium infection.

The role of macrophages in the innate immunity of (CBA/N female X DBA/2N male)F1 female mice to Salmonella typhimurium was assessed with silica, an agent which has been reported to selectively inactivate macrophages. Silica, administered intravenously to mice, markedly decreased the phagocytic capacity of splenic macrophages but had no effect on splenic responsiveness to the B-cell mitogen lipopolysaccharidide or the T-cell mitogen phytohemagglutinin, nor did it affect the frequency of surface immunoglobulin-positive cells (B cells). Silica given to mice 1 day before intraperitoneal challenge decreased the 50% lethal dose of S. typhimurium 100-fold. The incidence of survival of mice given silica up to 14 days before infection with a sublethal dose of organisms was also decreased. This susceptibility could also be demonstrated when silica was given 10 days, but not 20 days, after S. typhimurium infection. Poly-2-vinylpyridine-N-oxide, a lysosomal stabilizing agent, abrogated the silica effect. Deaths among silica-treated mice followed uncontrolled multiplication of the organism in the spleen. These results provide direct evidence that macrophages play an essential role in natural immunity to murine typhoid and demonstrate the efficacy of silica as a tool to analyze macrophage function.


Persistent infection of human lymphoid and myeloid cell lines with herpes simplex virus.

Herpes simplex virus (HSV) type 1 replicated and persisted in human T, B, and myeloid cell lines with different patterns of viral replication and various effects on cell growth. T cell line CEM supported the replication of HSV for over 400 days without detectable differences in cell growth as compared with uninfected cells. HSV persisted in B cell line NC37 and myeloid cell line K562 for up to 222 and 374 days, respectively, but led to a significant decrease in the number of viable cells by 7 weeks of infection. The average number of cells producing infectious virus was very low in these cell lines (range, 0.5 to 2.7+) compared with a larger proportion of cells exhibiting HSV antigens by immunofluorescence (range, 24 to 58%). In contrast, null cell line LAZ 221 failed to replicate HSV even though the viral infection led to a cessation of cell growth.


Depression of bovine monocyte chemotactic responses by bovine viral diarrhea virus.

Incubation of bovine peripheral blood monocytes with bovine viral diarrhea virus (either Singer or NY-1 strain) caused a consistent, statistically significant decrease in their random locomotion (no chemoattractant) and chemotaxis towards a chemotactic lymphokine. Chemotaxis was determined by a modification of the Boyden method. Incubation of bovine viral diarrhea virus with mononuclear cells depressed chemotaxis by a mean of 56% (P less than 0.0005). Heat-killed virus had no effect on monocyte motility. Data suggest that bovine viral diarrhea virus can rapidly suppress monocyte functions in vitro, but by unknown mechanisms, not by killing cells.


Type C virus and immunoglobulin A production by murine myeloma MOPC-315: two independent activities.

The suspected correlation between cessation of type C virus production and halt in immunoglobulin secretion by murine myeloma cells was studied. Employing two variants of the murine myeloma MOPC-315, immunoglobulin A-producing and nonproducing cells, we demonstrated that the two myelomas release similar levels of type C viruses which share common nucleotide sequences and that the viral genomes are equally expressed within the cells. Thus, the suggested relation between these two activities does not apply for MOPC-315 cells and probably for other murine myelomas also.


Isolation and development of a reticuloendotheliosis virus-transformed lymphoblastoid cell line from chicken spleen cells.

The establishment and characterization of a reticuloendotheliosis virus (strain T)-transformed lymphoblastoid cell line, designated TV-1, was reported. These cells, isolated from the spleen of a moribund chick infected with reticuloendotheliosis virus, were maintained in suspension culture for over 74 weeks at a stable generation time of 15 h. The cells were found to carry a female karyotype. Both TV-1 cells and cell-free TV-1 culture supernatant produced lesions and mortality patterns in chicks which were identical to those caused by reticuloendotheliosis virus (strain T) infection. Examination of TV-1 cells by electron microscope revealed the presence of C-type virions budding from the plasma membrane. Cytotoxicity assays and fluorescent antibody tests indicated the presence of B-cell determinants on the TV-1 cell surface membrane.


Cytolytic activity of macrophages isolated from primary murine sarcoma virus (MSV)-induced tumors.

Macrophages isolated from regressing or progressing tumors induced by murine sarcoma virus (MSV) were tested for cytolytic activity in a 18-h 51Cr release assay. Macrophages from the tumors of mice injected 14 days earlier with stocks of MSV-producing regressor or progressor tumors had comparable levels of cytotoxicity. However, macrophages from the progressively growing tumors, 50--65 days after the inoculation with the progressor virus, had lower levels of cytotoxicity than those from the regressing tumors (day 14). On the other hand, macrophages from progressively growing MSV tumors (day 14) in nude mice had little or no detectable cytolytic activity. In a fractionation study, using the 1-g velocity sedimentation technique, cells from regressing tumors (day 14) showed at least two peaks of cytolytic activity, one at about 4 mm/h and another at 6--7 mm/h. In contrast, none of the cell fractions of tumors from nude mice (day 14) showed cytolytic activity. Since bacterial lipopolysaccharide (LPS) has been shown to augment the cytolytic activity of primed macrophages, its effect on cells from MSV tumors was evaluated. Cytolytic activity of the cells from regressing or progr-ssing tumors that had pre-existing cytolytic activity was augmented by the addition of LPS during the assay period, but LPS treatment did not activate inactive fractions or change the distribution patterns of the cytolytic activity in fractions from 1-g velocity sedimentation.


Role of interferon in the pathogenesis of virus diseases in mice as demonstrated by the use of anti-interferon serum. VI. Polyoma virus infection.

We have determined the effect of a single injection of potent sheep anti-mouse interferon globulin on polyoma-virus-induced early runting disease and tumor formation in Swiss mice. When newborn mice were injected with greater than or equal to 9 X 10(6) PFU of polyoma virus, 16% (7/44) of mice runted and died in contrast to 96% (45/47) of mice injected with virus and anti-interferon globulin. Likewise, a greater percentage of mice injected with lower doses of virus and anti-interferon globulin developed tumors than did control virus-injected mice. These results suggest that interferon is an important factor in determining the susceptibility of newborn mice to polyoma virus.


Biological and biochemical observations on isolates of EB virus from the malignant epithelial cells of two nasopharyngeal carcinomas.

Foetal, adult seronegative, and cotton-top marmoset lymphocytes have been transformed into cell lines by EB virus from the malignant epithelial cells of two nasopharyngeal carcinomas. The nature of the cell lines was checked by karyotyping and by light and electron microscopy, and the presence of the EB virus genome was demonstrated by immunofluorescence tests. Immunofluorescence also showed the incidence of EB virus-producing cells in each line and this was checked by electron microscopy and the use of a lymphocyte transformation assay. Two foetal-derived lines did not produce virus spontaneously, could not be activated with various inducers, and were found by DNA reassociation kinetics to carry only small numbers of genome copies per cell. An adult-derived line produced virus, could be activated to produce more, and provided enough infectivity to transform marmoset cells. The resulting marmoset line made profuse transforming virus; it thus provides the first abundant source of NPC-derived infectious EB virus for comparative studies. The results are discussed in relation to the interactions with comparable target cells of EB virus from normal individuals and from patients with other diseases.


Effect of sodium butyrate on the antiviral and anticellular action of interferon in normal and MSV-transformed cells.

Sodium butyrate enhances the antiviral and anticellular action of interferon in MSV-transformed cells but has no such effect in normal cells. The increased sensitivity could be mediated at least partially by the butyrate-induced elaboration of the microtubule and microfilament network.


Tritiated thymidine as a broad spectrum initiator in transplacental two-stage carcinogenesis, with phorbol as promoter.

A single subcutaneous injection of 200 mu Ci [3H] thymidine into pregnant BALD/c mice, followed by intraperitoneal injections of phorbol, twice weekly for 25 weeks, in the offspring, resulted in higher tumour development in the lungs and livers of male and, to a lesser extent, of female offspring, than in their untreated littermates. The difference in overall tumour incidence was statistically significant, but the increases of the individual tumour types were only of borderline significance. Slight carcinogenic activity of [3H]thymidine alone was observed in the mothers, and in the offspring, without phorbol treatment. The results suggest the possibility of using [3H]thymidine as a broad spectrum initiator for transplacental two-stage carcinogenicity studies to determine the organ specificity of different promoting agents.


Absence of ecotropic or recombinant murine leukaemia virus in preleukaemic and leukaemic X-irradiated NZB mice.

NZB mice X-irradiated with a single dose of 630 R when they were 1-month old developed a high incidence of histologically defined lymphocytic leukaemia 8--25 weeks later. We have screened for murine leukaemia viruses (MuLV) in the lymphoid tissues of 8 of these leukaemic mice, and in 8 "preleukaemic", apparently healthy NZBs killed 1 month post irradiation. Xenotropic, but not ecotropic or recombinant MuLV, was detected by in vitro co-cultivation of bone marrow, spleen and thymus with selectively permissive cell lines, followed by the immunofluorescence test for MuLV gs antigen, and the XC test. Our results are not consistent, therefore, with the concept that the factor causing the leukaemias was an oncogenic virus activated by X-irradiation.


The epidemiology of Entamoeba histolytica in a Nigerian urban population.

Microscopic examination of multiple faeces samples of 2 825 persons was carried out in the Metropolitan Lagos. Overall prevalence of Entamoeba histolytica was 11.2%. Prevalence increased rapidly in younger age groups and there were no real differences between males and females. Prevalence was high among families who ate together from the same plate, among those who ate with their fingers and among those who ate away from home. Prevalence was not associated with type of water supply but was seemingly influenced by storage of household supplies. A low infection rate was associated with the availability of water closets and toilet habit. No association was found between prevalence and standard of education but the rate of infection was increased among workers with high occupational interaction. Significant differences were found among different ethnic groups. The highest infection rate was recorded in the last month of the wet season.


Secondary structure prediction of anterior pituitary hormones. Lack of correlation between predicted values and circular dichroism data.

The secondary structures of human somatotropin, human choriomammotropin, ovine and porcine prolactin, human, ovine and porcine beta-lipotropin, human and ovine lutropin, human thyrotropin, human corticotropin, alpha-melanotropin and human beta-melanotropin have been predicted by the method of Chou & Fasman. Predicted contents of alpha-helix and beta-sheet do not correspond well with values estimated from circular dichroism spectra.


Further characterization of the ovine lutropin alpha and beta subunits prepared by the salt precipitation method.

The subunits of ovine lutropin prepared by acid dissociation and salt precipitation were characterized by end group analysis, tryptic peptide mapping, SDS gel electrophoresis and biological activity. No evidence of internal peptide cleavage was found in the alpha subunit. The subunits possessed low activity. The alpha and beta subunits recombined effectively to generate a complex that had full receptor binding activity and in vitro biological activity. The recombinants of subunits prepared by countercurrent distribution showed only 50% activity in both assays. The salt precipitation method alpha subunit could be completely reduced and reoxidized in the absence of denaturants. The reoxidized alpha subunit combines with the native beta subunit generating full activity. However, this recombined hormone tends to lose activity with time, suggesting that the reoxidation may not fully restore the native structur of the reduced alpha subunit. The native lutropin alpha subunit effectively combined with follitropin beta subunit generating complete follitropin activity.


Relationship of geniculate and occipital PGO waves and the effects of nonvisual sensory activity.

The nature of occipital PGO waves (REM) was addressed by comparing the occipital PGO waves measured at the marginal gyrus with the geniculate PGO waves and the occipital PGO waves "directly" transmitted to the occipital cortex. The directly transmitted PGO waves lack the first short positive-going spike that precedes the negative phase of the occipital PGO waves. The "directly" transmitted PGO wave was recorded from cats with either deafferented or excised LG nucleus. The "directly" transmitted PGO wave resembled pontine negative-going PGO wave more regarding amplitude and wave shape changes than the occipital PGO waves of intact cats. The ascending negativity of pontine PGO waves is indirectly transmitted through phasic increase of the local excitatory processes, more in the LG than in the occipital cortex. The PGO waves seem to be affected by endogenous and exogenous nonvisual processes, more in the LG than the occipital cells alone. The occipital PGO waves seem to result from the vectorial sum of geniculate and "directly" transmitted occipital PGO waves.


Reversible binding of vibrio cholerae toxin on NAD-sepharose.

When a crude or partially purified culture of Vibrio Cholerae is passed through a column of NAD-Sepharose, the skin toxic material is bound to the gel. Choleragen may be recovered by increasing the ionic strength of the elution buffer, as shown by the skin test, immunodiffusion against anti-choleragen antiserum and polyacrylamide gel electrophoresis. Affinity chromatography on NAD-Sepharose may be helpful for the concentration and purification of Choleragen.


[Organ preserving surgical treatment of solitary and bilateral renal tumors].

The surgical treatment of patients with tumours in solitary kidneys and bilateral renal tumours is controversial. Conservative surgical procedures by excision of the tumour in situ or by extracorporeal "work bench surgery" with subsequent autotransplantation are a real alternative to radical nephrectomy which necessitates chronic dialysis or renal allotransplantation. 23 patients with tumours in solitary kidneys resp. bilateral renal tumours have been treated by local excision in situ (10 patients) or extracorporeal surgery (15 patients). The preoperative diagnostic investigations, indications for in situ resp. extracorporeal surgery, details of the operative procedure and results of treatment are presented and discussed.


[Acute emphysematous cholecystitis].

Emphysematous cholecystitis is a rare form of acute cholecystitis, characterized radiographically by the presence of gas within the gallbladder. We report of a patient, who was admitted to the hospital with the diagnosis of acute abdomen. This patient had an emphysematous cholecystitis caused by Clostridium perfringens. We found the wall of the gallbladder emphysematous and gangrenous, the gallbladder was distended and contained purulent material, but no stones. However, in addition, the films of abdomen showed gas in the ducts. Diagnosis, pathogenesis and the aetiological and therapeutical aspects will be discussed.


Electron microscopic studies on the endometrium of the grey seal (Halichoerus grypus) during its preparation for nidation.

The uterine luminal and glandular epithelia of two mature grey seals, a species with obligate delayed implantation, have been examined. Earlier studies of their corpora lutea had suggested that these females were in the pre-nidatory condition. Ultrastructural features of these epithelia in the animal which possessed the more mature corpus luteum indicated that the luminal epithelium had attained a state of differentiation similar to that of the pre-attachment phase in the laboratory rat, a state known to be compatible with nidation. The animal with the less well developed corpus luteum demonstrated the early stages of the early endometrial response to progesterone (the hormone which takes the uterus out of the delay condition). In the grey seal, despite the late start, a similar sequence of morphological changes to those present in the endometrium of normally implanting animals occurs. Thus, it seems that in the majority of eutherian mammals, endometrial preparation for nidation follows a relatively standard pattern, and occurs pari passu with the differentiation of the luteal tissue.