Comparison of 85Sr, 87mSr, 18F, and 99mTc-labeled phosphates for bone scanning. The observation by Subramanian and his co-workers that a 99mTc-labeled polyphosphate had excellent affinity for bone has led to widespread use of 99mTc-labeled phosphates as bone scanning agents. Initially, only polyphosphate was employed, but because of somewhat inconstant results and difficulty in preparation of this product, other phosphate compounds were sought. We soon discovered that an inorganic compound, pyrophosphate, appeared to have certain advantages over polyphosphate. Other workers formulated diphosphonates (organic phosphates) which also demonstrated advantages over polyphosphates. Comparison studies in rabbits utilizing 85Sr, 87mSr, 18F, and several phosphates (inorganic and organic) proved the 99mTc-labeled phosphates to be clearly superior in delineating normal skeletal anatomy. Studies in humans confirmed that excellent visualization of bone was obtained with 99mTc-labeled phosphates using either a gamma camera or a rectilinear scanner. What was not known, however, was just how reliable this class of agents would prove to be in detecting bone disease when compared to bone-seeking radiopharmaceuticals such as 85Sr, 87mSr, and 18F. Further comparative analyses have clearly demonstrated that both inorganic and organic 99mTc phosphate complexes are extremely sensitive in revealing more bone disease than the older bone scanning agents.


Biochemical anomalies of the nephrotic syndrome. The nephrotic syndrome is the only hypoalbuminaemic state frequently associated with hyperlipidaemia. In the presence of a negative nitrogen balance, hyperlipidaemia is metabolically inappropriate and reflects the result of persistent breakaway from free fatty acid control. This lipid abnormality may result in the premature development of ischaemic heart disease in patients in whom it is not possible to control the primary renal abnormality. The authors suggest that future work should be directed towards thyroxine and insulin metabolism in nephrotic states.


Diagnostic fiberoptic bronchoscopy: Techniques and results of biopsy in 600 patients. Six hundred patients underwent diagnostic flexible fiberoptic bronchoscopy (FFB). The two diseases most frequently encountered were bronchogenic carcinoma in 330 patients (55 percent) and bacterial infection in 94 (16 percent). A positive cytology on biopsy material was obtained in 279 of 330 patients (85 percent) with primary lung cancer. Fluoroscopy was a valuable aid in diagnosing bronchogenic carcinoma, since 42 percent of the tumors were not visible endoscopically and required fluoroscopic control for placement of the biopsy instrument. Of the 55 patients with hemoptysis and negative chest x-ray films, nine (15 percent) had fiberoptically visible endobronchial carcinomas! In addition, two patients with carcinoma of the larynx and one with carcinoma of the nasopharynx were discovered. Transbronchial biopsy (TBB) in 68 patinets with diffuse and localized disease achieved an overall 69 percent diagnostic success, including a correct diagnosis in each of four patients with Pneumocystis carinii pneumonia. Brush biopsy provided additional valuable laboratory data in bacterial, mycobacterial and cytomegalovirsu infectious but had a poor yield in Pneumocystis infection. Complications as a result of forceps biopsy were minimal, except for brisk bleeding in six patients.


Myocardial and plasma levels of adenosine 3':5'-cyclic phosphate. Studies in experimental myocardial ischemia. Alterations in myocardial and plasma levels of adenosine 3':5'-cyclic phosphate (cyclic AMP) were studied following clamping of the aorta or coronary artery occlusion in 30 dogs. Plasma cyclic APM levels increased markedly after thoracotomy but returned to control levels two hours later. Complete arrest of aortic flow (clamping) induced a significant early increase in the myocardial cyclic AMP levels of all animals studied. No increase was noted following pretreatment with propranolol or sham-occlusion. After localized coronary occlusion, only modest and insignificant changes occurred in plasma cyclic AMP levels in anesthetized animals and also in conscious dogs. The present study suggests that adrenergically mediated changes in tissue cyclic AMP content are an early manifestation of both generalized and local myocardial ischemia, while the plasma cyclic AMP level is a relatively insensitive indicator of small coronary occlusions.


The current status of prostaglandins and the lungs. The prostaglandins are a group of pharmacologically potent lipid compounds whose role in pulmonary pathophysiology remains to be defined. There is not question that these compounds can induce constriction (PGF) or relaxation (PGE) of the smooth muscle in bronchi and pulmonary vessels. There is no question that the lungs are a major site of both the synthesis and degradation. These facts make speculation about the role of the prostaglandins in pulmonary disorders a tempting exercise. However, the current paucity of clinical data prevents any valid assessment of such speculation at this time and emphasizes the need for continued investigation of the role of prostaglandins in patients with lung disease.


Asymptomatic pulmonary granular cell tumor presenting as a coin lesion. Granular cell tumor is a rare, usually benign neoplasm of neurogenic origin first described in 1926. Only 46 lesions, including the present case, of the 500 reported cases of this tumor, originated within the tracheobronchial tree. The case reported here is distinctly unusual because of its asymptomatic presentation as a coin lesion on chest roentgenogram. Electron microscopic examination of the tumor is presented.


Antibacterial activity of tobramycin against gram-negative bacteria and the combination of ampicillin/tobramycin against E. coli. The antibacterial activity of tobramycin, gentamicin, erythromycin, cloxacillin, kanamycin, cephalexin, penicillin, carbenicillin and polymyxin were compared against 303 clinical bacterial isolates from a pediatric hospital patient population. Standard disk diffusion and agar-dilution methods were employed. Significant activity was demonstrated for tobramycin against pseudomonas, Klebsiella, Escherichia coli and both Staphylococcus aureus and albus; Tobramycin was significantly more active against Pseudomonas than gentamicin or the other antibiotics testedmcomparable activity to gentamicin was present for the other types of bacteria; Cross-resistance was not encountered between tobramycin and gentamicin. 30 isolates of E. coli were tested against the combination of tobramycin and ampicillin by the growth-curve method. Synergism was demonstrated in 4 isolates, antagonism in 1 and an additive effect in 25. A bactericidal effect was present at 24h against 17 isolates with tobramycin alone and against 25 isolates when combined with ampicillin. These results provide in vitro rationale for the consideration of tobramycin for clinical use in patients with Psuedomonas infections for the combination of ampicillin and tobramycin for the treatment of selected E.coli infections.


[Verner-Morrison syndrome].

[Verner-Morrison syndrome]. Based on one case the Verner-Morrison-syndrome is demonstrated and discussed in detail. Clinically profuse watery diarrhea and severe hypokalemia are predominant. The pathogenesis is not quite clear. Curative therapy consists in radical surgical removal of the endocrinologically active tumores or diffuse hyperplasia.


Protein phosphorylation and metabolic control. The regulation of enzyme activity through the phosphorylation and dephosphorylation of specific seryl or threonyl residues in enzymes is now recognized as an important control mechanism. A great many non-enzymic proteins may also be interconverted between phosphorylated and non-phosphorylated forms, but in these instances the function served by phosphorylation is not well understood. This lack of understanding is probably due to our lack of knowledge of the specialized actions of most non-enzymic proteins. The sequences of phosphorylation and dephosphorylation of proteins are, of necessity, subject to rigid control. If this were not so, the ATP supply of a cell would be rapidly depleted. Moreover, such processes must be regulated for metabolic interconversions to have a physiological regulatory role. For the best studied system, the interconversion of phosphorylase b and phosphorylase a, many different factors controlling the phosphorylation and dephosphorylation steps have been elucidated. Probably this process is constrained so that it uses only a little energy. In this paper, these constraints are examined.


Energy utilization for control.

Energy utilization for control. When, on addition of a suitable substrate, a chemical potential is applied to an enzymic process such as glycolysis or respiration, whether in solution or membrane-bound, all components of the process pass into a nonequilibrium state, which might be steady or non-steady and which produces the following phenomena: (1) The reactants of each enzymic reaction are displaced from their equilibrium concentration, and energy is dissipated; (2) Part of each enzyme is transferred to a transition state of its catalytic function as well as isosteric and allosteric controlling functions, displaying local and gross conformation changes, and a rate-controlling state is generated; (3) In cyclic portions of a process futile events and chemical interconversion may occur; (4) In self- and cross-coupled portions of a process, oscillation with periodic changes of states and spatial propagation as well as instabilities may be observed; (5) At each step of a process, depending on the rate of flux and the specific enzymic function, a varying proportion of the free energy changes--which are concentration-dependent and derived from the overall potential of the system-is contributed to the control of flux rates. This will be exemplified for enzymes of bioenergetic pathways.


Magnetic resonance studies of specificity in binding and catalysis of phosphotransferases. Two common characteristics of the active site structures of intermediate complexes formed in kinase reactions have been observed by magnetic resonance techniques. First, in creatine, arginine, adenylate and pyruvate kinases (EC2.73.2, 2.7.3.3, 2.7.4.3 and 2.7.1.40, respectively) water is progressively excluded and the structure at the active site is progressively immobilized as each reactant is successively added to the enzyme, as monitored by electron spin resonance (e.s.r) and the enhancement of the proton relaxation rate of water (PRR) due to paramagnetic manganese(II) probe. Significant, and often wide-spread, changes in the protein conformation accompanying successive additions of reaction components are shown with 1H n.m.r. studies of pyruvate kinase. The second characteristic is that, for the ternary enzyme-Mn-nucleotide complexes, two parameters, the e.s.r. spectrum and PRR enhancement values, fall within a range of 10% for all enzymes investigated, with the exception of bovine brain creatine kinase. These similarities suggest a homology in teriary structure at the active sites of these enzymes. An unsuspected aspect of substrate and cofactor specificity has been revealed by e.s.r. spectroscopy of the manganese(II) complexes of the transition-state analogue of creatine kinase (E-MnADP-formate-creatine) and of the ternary phosphoenolpyruvate complex. In the former case, replacement of ADP, the normal substrate, by its substrate analogues IDP or 2acute-deoxyadenosine diphosphate produced two interconvertible species of the transition-state analogue complexes, observed in the e.s.r. spectra as an isotropic species and a highly anisotropic species. With the normal substrate, only the anisotropic species is observed. Similarly, in the case of the complex pyruvate kinase-Mn-phosphoenolpyruvate, when the normal monovalent activator K+ is replaced by the inert tetramethylammonium ion, again two interconvertible species rather than the normal one species are observed by e.s.r. spectroscopy. The implications of these phenomena for the relation of specificity to catalytic efficiency are discussed.


[Increased cAMP concentration in parotid secretion in arterial hypertension: relationship to plasma-renin activity (author's transl)]. Hypertensives with normal and increased plasma-renin activity have an increased salivary excretion of cyclic adenosine monophosphate (cAMP) such as can be obtained in normal persons by stimulation of beta-adrenergic fibres. Propanolol, as a beta-receptor blocker, reduces such increased cAMP levels in hypertensives to minimal levels. The results suggest that beta-receptors are stimulated in hypertensives with normal or increased renin activity.


[Acute polyneuropathy caused by diphenylhydantoin intoxication (author's transl)]. In a 34-year-old epileptic male who had been treated for more than ten years with an average of 300 mg diphenylhydantoin and 200-300 mg phenobarbitone daily severe status  epilepticus occurred which could only be controlled with high doses of diphenylhydantoin and phenobarbitone intravenously. During this treatment a severe mainly motor polyneuropathy occurred acutely which was more pronounced in the distal parts of the legs, and cerebellar symptoms were noted at the same time. The neuropathy largely regressed during the following year whereas the cerebellar symptoms persisted. There is evidence for direct toxic damage of peripheral nerves due to diphenylhydantoin. Phenobarbitone in high doses inhibits degradation of diphenylhydantoin competively. In this patient it very likely contributed towards the development of sever intoxication leading to neurological defects.


The physicochemical properties of the cytoplasmic androgen receptor in the kidneys of normal, carrier female (tfm/+) and androgen-insensitive (tfm/y) mice. The physicochemical properties of the androgen-receptor complex in mouse kidney were determined. The sedimentation coefficient, 7.9S and Stokes radius of 82A, are compatible with an asymmetric protein [frictional ratio f/fo 1.98; axial ratio, assuming a prolate ellipsoid, 20] having a molecular weight of 270,000 daltons. The kidney receptor is a relatively acidic protein of esoelectric point (pI) 4.8 and readily precipitable with protamine sulfate or ammonium sulfate. Studies with protein-specific reagents suggest that both cysteine and tryptophan residues may be necessary for maintaining the functional configuration associated with androgen binding. The kidney receptor can promote the association of testosterone with purified DNA. These properties of the androgen receptor in mouse kidney are remarkably similar to those of male accessory sexual tissue. The receptor detected in carrier female mice (tfm/+ heterozygous for the gene for testicular feminization (tfm), has the same physical properties as that of normal mice. However, due to a decrease in receptor concentration, binding activity is only 69% that of normal. In cytosol from androgen-insensitive mice (tfm/+), a specific androgen receptor cannot be demonstrated by 5 different techniques.


Guanidination of ovine luteinizing hormone and effects on activity. The free amino groups in oLH, oLHalpha and oLHbeta were guanidinated by O-methylisourea. The epsilon-NH2 groups of lysine residues reacted bo substitute these positions in the sequence with the more basic homoarginine residue. The alpha-NH2 groups did not react under the conditions used. Guanidinated oLH or the products of guanidinated oLHalpha + native oLHbeta or guanidinated oLHalpha + guanidinated oLHbeta were inactive in two bioassay systems. Native oLHalpha + guanidinated oLHbeta, however, showed potencies of 39% to 55% of that observed with the native subunit recombinant or native oLH. Possible structural implications for hormone-receptor site interactions are discussed.


Adrenal cells in tissue culture the effects of choleragen and ACTH on steroid and cyclic-AMP metabolism. Primary cultures of mouse adrenocortical tumors provide a sensitive system for investigating the effects of the enterotoxin of the V. cholerae (choleragen) on cyclic-AMP metabolism in the intact cell. Like ACTH, the toxin stimulates the synthesis and release of steroids from these cells but its mode of action differs from that of ACTH. The steroidogenic response to ACTH is immediate and of limited duration. The initial rate of steroidogenesis is the highest. In contrast, the steroidogenic response to choleragen is preceded by a 30-240 minute lag period which is inversely related to the concentration of the toxin. Whereas prolongation of the response to a single dose of ACTH requires hormone concentrations above those producing maximal initial steroidogenic activity, persistent steroidogenesis is induced at all levels of the toxin. Steroidogenic responses are detectable with 10 pg/ml of choleragen or less. The respective effects of ACTH and choleragen on cyclic-AMP synthesis and release into the medium parallel those on steroidogenesis. Intracellular cyclic-AMP levels in ACTH-treated cells reach a peak within 20-30 minutes and decline to normal levels within 2-4 hours. In choleragen-treated cells, after the lage period, the levels of intracellular cyclic-AMP remain above control levels indefinitely. The effects of ACTH and choleragen on cyclic-AMP biosynthesis are additive at all levels of the two compounds. The effects of choleragen are blocked by prior treatment of the toxin with a five-fold molar excess of ganglioside GM1, a presumed constituent of the toxin-binding site.


Lipolytic action of human chorionic somatomammotropin. Human chorionic somatomammotropin extracted and purified from placenta at term was proved to have a lipolytic action in the epididymal fat pad of rats. The following mechanism appears to be involved in the lipolytic action of the hormone; human chorionic somatomammotropin activates adenyl cyclase, thereby increasing the concentration of cyclic AMP in the tissue, which, in turn, activates protein kinase to lead to the activation of hormone sensitive lipase.


The effects of cyclic AMP, theophylline, angiotensin II and electrolytes upon renin release from rat kidney slices. The effects of cyclic AMP, theophylline, angiotensin II and electrolytes upon renin release were examined by incubation of rat kidney slices.  Angiotensin inhibited renin release with increasing concentrations.  On the other hand, cyclic AMP and theophylline stimulated it.  Calcium also seemed to play an important role in the control of renin release from kidney slices.  However, the direct effects of sodium and potassium upon renin release were not conspicuous.


A case report of Turner's syndrome with ring X chromosome. A case of Turner's syndrome with short stature and 45, XO/46, XXr mosaicism in chromosome study was presented. With special emphasis on endocrinological study, the size of the breasts was normal in contrast to the poor development of the breasts in most of types of Turner's syndrome. She showed normal thyroid function, slightly low level of urinary 17-OHCS, decreased 17-KS, poor response in metopirone test and poor response of HGH to insulin.


Qualitative abnormality of insulin secretion in a case with insulinoma. We have presented here a case of atypical insulinoma.  Despite the recurrent episodes of hypoglycemic symptoms, the plasma level of insulin has never been excessive at fasting or by regular provocative tests.  Detailed examination had demonstrated qualitative abnormality of insulin secretion.  Hyposuppressibility of insulin secretion by hypoglycemia, borderline diabetic curve of glucose tolerance test, blunted response ot insulin to glucagon and leucine were the principle characteristics of these abnormalities.  After removal of adenoma, insulin response to glucose, glucagon and leucine was improved.  Only secretion provoked a high level of insulin and this abnormal elevation was no longer seen after the removal of adenoma.  A removed elevation was no longer seen after the removal of adenoma.  A removed insulinoma contained 25 U of immunoreactive insulin per gram tissue, but was negative for aldehyde-fuchsin staining.  On electromicroscopy only atypical beta-cell granules were seen.


A case of acromegaly improved by pituitary apoplexy. This report deals with a detailed course of one patient with acromegaly who had a pituitary apoplexy.  The pituitary apoplexy occurred suddenly 5 days after administration of a oral hypoglycemic agent, buformin, during hospitalization.  Immediately after the attack changes of the concentrations of several hormones such as serum growth-hormone, serum thyroid hormone and urinary 17-hydroxycorticosteroids were followed until the development to hypopituitary state.  Simultaneously with the decrease of the concentrations of the above-mentioned hormones, a regression of the physical manifestations of acromegaly and a complete amelioration of diabetes mellitus were observed.


Reserpine induced changes in the estradiol uptake by the pituitary and the uterus in ovariectomized rats. Reserpine treatment produced a marked decrease of the in vitro binding of estradiol to the nuclear fraction of the uterus. A similar observation was made when reserpine was given into the incubation medium of uterine tissue. In contrast to these findings both in vitro and in vivo reserpine administration resulted in an increase of the estradiol binding to the nuclear fraction of the anterior pituitary in ovariectomized rats. The binding capacity of cytosol fraction after reserpine administration did not show significant alterations. In addition to these observations it is worth to mention that reserpine treatment resulted in a significant decrease of the estradiol-induced increase of uterus weight in ovariectomized rats.


Effects of temperature on interictal discharge at penicillin epileptogenic foci. The effects of local brain temperature on acute focal penicillin epilepsy in the exposed hippocampus of cat were studied. Results from anesthetized and from immobilized, unanesthetized animals were compared. Over the temperature range 26 to 43 degrees C (at the alveus), the interictal spike interval and duration of the spike discharge varied inversely with temperature. The former showed a Q10 of 2.4 and the latter a Q10 of 1.5, with no difference due to type of preparation. A significant trans-hippocampal thermal gradient may imply that these values are underestimates by 20% or more. The low Q10 of duration of the paroxysmal discharge was consistent with the known temperature dependence of impulse conduction velocity of intracortical neural networks. The high Q10 of the interictal interval, on the other hand, was consistent with the view that some slow endogenous, perhaps metabolic factor such as a NA,K-ATPase modulated excitability at the focus of penicillin spikes.


Attempts to produce spike-and-wave complexes in the electrocorticogram of the rat. The effects of several procedures for inducing spike-and-wave complexes in other experimental animals have been examined in the rat. Systemic convulsants, pentylenetetrazol, methionine sulfoximine and fluoroacetate, topical picrotoxin, and bilateral implants of cobalt produced the appearance of polyspike discharges in the ECoG. Topical application of conjugated estrogens to both frontal cortices elicited single spike-and-slow-wave complexes which did not repeat at 3 to 5 cps. These results suggest that the rat is not a suitable species for a detailed study of spike-and-wave production.


Oxidation of succinate in heart, brain, and kidney mitochondria in hypobaria and hypoxia. Exposure of rats to hypobaric stress for periods of up to 36 h caused a consistent change in the succinate-NT reductase activity of the heart mitochondria whereas there was no significant change in the activities of either succinate dehydrogenase and succinate-NT reductase of the brain and the kidney. Mitochondrial succinate dehydrogenase of the heart, the brain and the kidney was activated 2- to 7-fold with the substrate and malonate. The activations obtained with oxalate, citrate and dinitrophenol were relatively lower in comparison to succinate and malonate. Benzohydroquinone and 2-nitrophenol had no stimulatory effect on the heart, the brain and the kidney mitochondria. THE ACTIVATIONS OBTAINED WITH THE VARIOUS EFFECTORS PARTIALLY (OR COMPLETELY IN THE CASE OF SUCCINATE) REVERSED ON WASHING THE MITOCHONDRIAL SAMPLES WITH THE SUCROSE HOMOGENIZING MEDIUM. The effect of ubiquinol, which also activated the enzyme, was only partially reversed after the second preincubation with succinate in the brain and the kidney whereas in the heart the activity was fully reversed. The increased activity of succinate dehydrogenase obtained with ATP and ADP was further enhanced by Mg2+ exclusively in the brain mitochondria, suggesting the possibility of Mg2+-AIP complex as the active species. Succinate-NT reductase of the heart, the brain and the kidney mitochondria showed a high activation with ubiquinone whereas its reduced form had no stimulatory effect.


Regulatory properties of purified 3-phosphoglycerate dehydrogenase from Bacillus subtilis. 3-Phosphoglycerate dehydrogenase (3-phosphoglycerate:NAD oxidoreductase, EC. 1.1.1.95) was purified from Bacillus subtilis by conventional methods. The final preparation was homogeneous by electrophoretic analysis and had a sedimentation constant of 6.3 S. On the basis of gel filtration data the enzyme had a molecular weight of about 166000. The plot of velocity versus phosphoglycerate concentration was biphasic while similar plots for hydroxypyruvate phosphate and NADH were the conventional hyperbolic type. The enzyme was specifically inhibited by serine. The inhibition was time dependent, requiring several minutes incubation before a constant level of inhibition was achieved. Serine inhibition was of the "mixed type" with respect to 3-phosphoglycerate and Hill plots of these data had slopes that approached 2. Desensitization of the enzyme to serine inhibition was achieved by incubation in the absence of dithiothreitol. The desensitized enzyme was different from the native enzyme in fluoresence properties, sedimentation characteristics and in the absence of the biphasic phosphoglycerate saturation curve. Evidence was obtained for the participation of sulphydryl groups in the changes in protein structure responsible for serine inhibition as well as the dehydrogenase activity of the enzyme.


Effects of triiodothyronine on rat-liver mitochondrial transcription process. 1. In order to demonstrate that triiodothyronine affects mitochondrial RNA synthesis by acting on the enzyme component of the DNA. RNA polymerase complex, mitochondrial RNA polymerase from thyroidectomized and hormone-treated rats was purified up to a stage in which activity was dependent on the addition of exogenous template. In these conditions and using different DNAs as templates, the enzyme from hormone-treated animals displayed an activity about double that of the activity of thyroidectomized animals. 2. Measurements of stability of mitochondrial RNA synthesized in vitro suggest, however, that the hormone can act also at the template level in mitochondrial transcription: the RNA population synthesized in vitro from hormone-treated rats is indeed much more enriched in unstable, probably messenger, RNA species. 3. The turnover of mitochondrial messenger RNA is higher after hormone treatment. 4. Adenosine cyclic 3':5'-monophosphate (cAMP) and its dibutyryl derivative added in vitro to mitochondria from thyroidectomized animals do not affect the incorporation of labeled precursor into mitochondrial RNA, suggesting that the level of the cyclic nucleotide in mitochondria is probably not involved in the hormone action. 5. It is concluded from these and previous studies that the thyroid hormone affects more than one parameter in the mitochondrial transcription process. The interrelationship between these events at molecular level remains, however, to be clarified.


Existence of two alternative pathways for fructose and sorbitol metabolism in Bacillus subtilis Marburg. Strains of Bacillus subtilis mutated for fructose phosphotransferase system (fruA), fructose-1-phosphate kinase (fruB), fructokinase (frucC) have been tested for their catabolism of sorbitol and fructose. It is shown that the previously known pathways of sorbitol and fructose degradation in B. subtilis, e.g.: (see article) may metabolize intracellular fructose produced either by sorbitol oxidation or by fructose-1-phosphate dephosphorylation. The intracellular fructore degradation via fructose-1-phosphate kinase has been found to require the fructose phosphotransferase system which ensures a vectorial transport of fructose.


The mechanism of action of methionyl-tRNA synthetase from Escherichia coli. Inhibition by adenosine and 8-aminoadenosine of the amino-acid activation reaction. Adenosine and 8-aminoadenosine, both competitive inhibitors of ATP-Mg2+ in the ATP-PPi exchange reaction catalyzed by methionyl-tRNA synthetase, are used to investigate the active center for methionyl-adenylate formation. Resolution of the kinetics parameters of the reaction indicates that methionine markedly enhances the affinity of the nucleosides for the enzyme, providing evidence for coupling between the sites for amino acid and the nucleoside moiety of ATP. Furthermore, occupation of both of these sites is a prerequisite for binding of pyrophosphate. Introduction of an amino group in position 8 of the adenine ring strongly increases the affinity constants for the nucleoside and for pyrophosphate in the coupled reactions described above.


Chlorothricin, and inhibitor of porcine-heart malate dehydrogenases, discriminating between the mitochondrial and cytoplasmic isoenzyme. The macrolide-type antibiotic chlorothricin was found to inhibit both the mitochondrial and the cytoplasmic form of pig heart malate dehydrogenase. Steady-state kinetic measurements revealed that in the direction of oxalacetate reduction chlorothricin is competitive with respect to NADH and non-competitive with respect to oxalacetate. Both the variation of initial velocity with NADH concentration in the presence of antibiotic, and, at several fixed levels of NADH, the variation of initial velocity with chlorothricin concentration deviates from the classical Michaelis-Menten relationship for the two isoenzymes. Since, despite the very similar kinetic behaviour of the mitochondrial and cytoplasmic species of malate dehydrogenase, the concentration of chlorothricin required for half-maximal inhibition of the two enzymes differs by more than a factor of 10 (the mitochondrial isoenzyme being more susceptible to inhibition), it is concluded that the NADH binding sites of the mitochondrial and cytoplasmic form of malate dehydrogenase from pig heart are different.


Deterioration of rat-liver mitochondria during isopycnic centrifugation in an isoosmotic medium. We have investigated the effect of the centrifugation speed on the behavior of rat-liver mitochondria during isopycnic centrifugation in an isoosmotic medium. The gradient was made with a macromolecular compound, glycogen dissolved in 0.25 M aqueous sucrose. The distribution curves of several mitochondrial enzymes change when the centrifugation reaches a certain speed: they are shifted toward regions of lower density. The results are plausibly explained by supposing that the inner mitochondrial membrane becomes permeable to sucrose at high centrifugation speeds, and that the granules swell. The main causal agent of the phenomenon is the hydrostatic pressure the mitochondria are subjected to during centrifugation. Morphological observations show that mitochondria are markedly deteriorated when centrifuged at high speed in the glycogen gradient; they are swollen and the outer membrane is broken; also frequently, a large electron-dense granule is seen in the matrix near the inner mambrane.


Blocks in elongation and initiation of protein synthesis induced by interferon treatment in mouse L cells. Synthesis of polypeptide chains coded by exogenous messenger RNAs is inhibited in cell-free extracts from interferon-treated mouse L cells, due to a "deficiency" in some specific tRNA species. A detailed analysis shows that polypeptide chain elongation is blocked and incomplete chains are formed. After a few minutes, however, initiation of new polypeptide chains is also blocked. Messenger RNA still binds to ribosomes but initiator Met-tRNA(FMET) binding is inhibited. The block in initiation appears to be secondary to the block in elongation.


Gylcerol-3-phosphate shuttle and its function in intermediary metabolism of hamster brown-adipose tissue. 1. Brown adipose tissue of the hamster possesses high specific activities of soluble, cytoplasmic NAD-linked, as well as mitochondrial flavin-coupled, glycerol-3-phosphate dehydrogenases. The ratio of the two enzyme activities is high (close to 1), when compared with other tissues of the hamster. 2. In the presence of rotenone, NADH is oxidised very poorly by homogenates of brown adipose tissue. A high rate of oxidation is obtained upon further addition of dihydroxyacetone phosphate, which itself is negligible oxidised. When followed fluorimetrically glycerol 3-phosphate can also be observed to induce NADH oxidation, but only after a significant lag time. Similar results are obtained with isolated mitochondria plus high-speed supernatant. With high-speed supernatant alone, only dihydroxyacetone phosphate has any effect, whereas with isolated mitochondria neither dihydroxyacetone phosphate nor glycerol 3-phosphate induce any NADH disappearance. 3. Respiration induced by NADH plus dihydroxyacetone phosphate in homogenates equals 56% of the respiration induced by glycerol 3-phosphate alone. 4. Respiration induced by NADH plus dihydroxyacetone phosphate, as well as that induced by glycerol 3-phosphate, is inhibited by the same concentrations of inhibitors as are required for inhibition of the mitochondrial dehydrogenase i.e. EDTA, long-chain unsaturated fatty acids, long-chain fatty acyl CoA esters. 5. In isolated brown adipocytes in the presence of rotenone, norepinephrine significantly inhibits respiration induced by glycerol 3-phosphate. 6. The results obtained are discussed with respect to the role of glycerol 3-phosphate as an electron sink for cytosolic reducing equivalents to maintain a low level of extramitochondrial NADH. A means of maintaining a level of glycerol 3-phosphate adequate for triglyceride synthesis is also considered.


Iron-sulfur components of succinate dehydrogenase: stoichiometry and kinetic behavior in activated preparations. Extensively or completely activated preparations of beef heart succinate dehydrogenase have been investigated by electron paramagnetic resonance (EPR) techniques at 6 to 97 K. Reductive titrations with dithionite and rapid kinetic studies were performed with various types of soluble and membrane-bound preparations of the enzyme. The following components were detected and their behavior analyzed: a free radical, presumably arising from the covalently bound flavin on reduction, two iron-sulfur centers of the ferredoxin type, the signals of which appear on reduction, and a highpotential iron-sulfur component, detectable in the oxidized state. The high-potential component was only detected in complex II and inner-membrane preparations. This component and one of the ferredoxin-type centers were present in amounts close to stoichiometric with the flavin and were reduced by substrate. The other ferredoxin-type center was present in amounts between 0.1 and 0.5 times that of the flavin and was reduced only by dithionite. Of the components reduced by succinate, however, only a fraction (up to 50% of the high-potential iron-sulfur center and 40-60% of the ferredoxin-type iron-sulfur center) was reduced within the turnover time of the enzymes; In complex II not more than about 10% of the flavin appeared in the semiquinone form at any time. Soluble, purified preparations behaved similarly except that the high-potential component was nearly or completely absent and extensive accumulation of the free radical occurred (up to 70 to 80% of the flavin) in titration and kinetic experiments. No significant difference was observed between the rates of semiquinone formation and the reduction of the ferredoxin-type or high-potential centers by the substrate. Also no qualitative differences in the properties studied in this work became apparent between prepatations containing 4 or 8 iron atoms, respectively.


Lysophospholipids in pig pancreatic zymogen granules in relation to exocytosis. A hypothesis to explain the stimulatory role of cyclic AMP (adenosine 3':5'-monophosphate) in pancreatic enzyme secretion. has been tested. In this hypothesis cyclic AMP would activate a phospholipase activity, which would lead to a locally increased lysophospholipid formation, resulting in a fusion between the zymogen granule membrane and the apical plasma membrane. Cyclic AMP added to isolated pig pancreatic zymogen granules leads to an increased lysis of these granules, but the slowness of this effect makes its physiological significance dubious. In pancreatic homogenates or zymogen granules no stimulating effect of cyclic AMP on lipase of phospholipase activity could be demonstrated. Isolated zymogen granules have a high lysophospholipid content (27% of total phospholipids), consisting of the 1-acyl and 2-acyl forms of lysophosphatidylcholine and lysophosphatidylethanolamine. Experiments with radioactive phosphatidylcholine indicate that the lysophospholipids are due to the action of endogenous (phospho)lipases during the isolation procedure. It is concluded that these experiments do not lend support to the above hypothesis for the mechanism of action of cyclic AMP in pancreatic enzyme secretion


Interactions of heteroaromatic compounds with nucleic acids. 2. Influence of substituents on the base and sequence specificity of intercalating ligands. This paper presents the results of a systematic study on the effects of substituents on the base and sequence specificity of tricyclic heteroaromatic compounds interacting with DNA by intercalation. All the compounds tested are derived from proflavine and acridine orange analogs with different heteroatoms in the middle ring. Their base and sequence specificities were determined by differential dialysis of the ligand against DNA samples of differing G-C content. The main results indicate that (a) the introduction of a phenyl substituent into one of the two available positions of the middle ring increases or decreases the G-C specificity of the ligand depending on the position where the substitution takes place; (b) compounds of the substitution type of neutral red (2-methyl-3-amino-7-dimethyl-amino-phenazine) show unexpectedly high G-C specificities and (c) DNA ligands of pronounced sequence specificity for adjacent G-C pairs can be constructed by combining the structural elements of neutral red with an additional phenyl residue in the same molecule. The further study of compounds related to the phenylated neutral red revealed that the G-C specificity can be improved or destroyed by additional substituents. The comparison of the G-C specificity and the DNA-affinity data of the compounds studied leads to the suggestion that the specificity arises mainly from electronic factors which are strongly controlled through steric constraints on possible ocmplex geometries. As a basis for the discussion a possible structure for the DNA complex of the phenylated neutral red is considered in which the extra phenyl ring at N-5 of the phenazinium system, protrudes into the large groove of the DNA helix while the tricyclic part of the ligand is inserted between the DNA base-pairs.


The isolation of nuclei and basic nucleoproteins from the cellular slime mold Dictyostelium discoideum. A method is described for the isolation of nuclei from an axenic strain of Dictyostelium discoideum using a sorbitol/Ficoll solution and low concentration of Triton X-100. Basic proteins have been extracted from the nuclei and on polyacrylamide gel electrophoresis yield a consistent pattern in which five major groups or bands predominate. Four of these five fractions comigrate with calf thymus histones and one fraction seems to be unique to D. discoideum. The slowest moving of the five fractions is soluble in 0.5 M perchloric acid and comigrates with calf thymus histone F1. After recovery from the perchloric acid solution by precipitation with acetone this fraction yielded one major band on electrophoresis.


Kinetic equivalence of the active sites of alcohol dehydrogenase from horse liver. The reduction, catalysed by liver alcohol dehydrogenase, of benzaldehyde in the presence and absence of pyrazole, and the oxidation of benzyl alcohol and cyclohexanol in the presence of isobutyramide, has been measured by the stopped-flow technique. In performing these experiments particular care was taken to purify the enzyme, coenzymes, substrates and inhibitors, and to minimise as much as possible the effects of a blank substrate reaction. The calculation of the amount of substrate converted to product during the various phases of the transient process was based on the absorption coefficients for the enzyme-coenzyme and enzyme-coenzyme-inhibitor complexes determined in the absence of substrate. The results show that the two active sites of liver alcohol dehydrogenase are kinetically equivalent and that the enzyme does not exhibit half-of-the-sites reactivity.


Ribosomal subunit localization of hemoglobin messenger RNA. When rabbit reticulocyte polyribosomes are treated with 0.5 M KCl, they dissociate into subunits and release a protein fraction which is required for peptide chain initiation in a cell-free system using KCl-treated subunits as the source of ribosomes. Three independent methods were used to determine the fate of mRNA after KCl treatment of the subunits. These three methods (sucrose gradient analysis of RNA after dissociating it from protein with sodium dodecylsulfate, acrylamide gel electrophoresis of RNA and electron microscopic analysis of subunits) all showed the 8--9-S mRNA to be associated with the small subunit, but not the large subunit. Furthermore, no mRNA was found to be associated with either "native" ribosomal subunit in a reticulocyte lysate.


The sources of plasma cyclic AMP: studies in the rat using isoprenaline, nicotinic acid and glucagon. The effects of intravenous administration of isoprenaline, glucagon and nicotinic acid on plasma concentrations of cyclic AMP in rats are described. In order to determine the relative importance of the liver as a source of extracellular cyclic AMP, the effects of the hormones were investigated in intact and functionally hepatectomised rats. The results showed that hepatectomy did not prevent an isoprenaline-stimulated increase in plasma cyclic AMP concentrations, although glucagon was without effect on plasma nucleotide concentrations in this group of animals. It is suggested that the liver is essential for the action of glucagon but that isoprenaline can increase plasma cyclic AMP concentrations in hepatectomised animals by increasing extrahepatic release of the nucleotide. Since inhibition of adipose tissue lipolysis with nicotinic acid did not prevent an isoprenaline or glucagon-stimulated increase in plasma cyclic AMP concentrations, adipose tissue is discounted as a major source of plasma cyclic AMP.


The effect of pregnancy on the control of lipolysis in fat cells isolated from human adipose tissue. 1. Lipolysis has been estimated by measuring the release of glycerol in isolated adipose tissue cells obtained from women in early prognancy, late pregnancy and 1 - 3 days post partum and from non-pregnant women. 2. Adipocytes of women at the end of pregnancy exhibited higher rates of lipolysis in response to adrenaline (1.5 - 15 muM) plus phentolamine (13 muM) than those of non-pregnant women or those in early pregnancy. 3. Lipolysis in response to adrenaline plus phentolamine in fat cells from women 1 - 3 days post partum was reduced compared to that at the end of gestation but was enhanced relative to that in the non-pregnant or early pregnant state. 4. Basal lipolysis also tended to be greatest at term. 5. Under conditions where the production of cyclic AMP was not rate limiting for the stimulation of lipolysis, that is in the presence of dibutyryl cyclic AMP (1 mM) or adrenaline (15 muM) plus phentolamine (13 muM) plus caffeine (1 mM), the release of glycerol in cells from women at term and in the puerperium was greater than that in women in the non-pregnant or early pregnant state. 6. Cell levels of cyclic AMP rose after incubation with adrenaline (6 muM) plus phentolamine or adrenaline (15 muM) plus phentolamine plus caffeine (1 mM) but were similar in all four groups of women. 7. It is concluded that the observed enhancement of lipolysis demonstrated in fat cells from women at the end of pregnancy reflects an increase in hormone-sensitive lipase activity rather than a modification of hormone receptor site sensitivity or of the rates of synthesis or breakdown of cyclic AMP. 8. This increase in adipose tissue lipolysis at the end of gestation could contribute to the reported rise in plasma nonesterified fatty acids in the final weeks of pregnancy.


Regional and systemic haemodynamic effects of some vasopressins: structural features of the hormone which prolong activity. Cardiac output and regional blood flows to myocardium, gut, uterus and kidney were determined in anaesthetised female rats by a single injection of 86RbCl. The haemodynamic responses were measured at various time intervals up to 2 h after single I.V. injections of lysine-vasopressin and the following of its analogues: a) with extended peptide chains at the N-terminal (including "hormonogens") Nalpha-glycyl-glycyl-lysine-vasopressin, Nalpha-glycyl-glycyl-glycyl-arginine-vasopressin and Nalpha-D-valyl-lysine-vasopressin, b) "carba" modifications desamino-carba6-arginine-vasopressin, desamino-carba6-D-arginine8-vasopressin, desamino-carba6-ornithine8-vasopressin, desamino-dicarba-arginine-vasopressin and c) other steric alterations - desamino-D-arginine8-vasopressin and desamino-N-methylarginine8-vasopressin. Sub-pressor doses of lysine-vasopressin were followed by marked reductions in gut and uterus blood flows which reached a peak at 10 min. and had completely receded by 60 min. The presence of steric alterations in the C-terminal tripeptide of the molecule- D-arginine or N-methylarginine in sequence position 8 - practically completely eliminated vascular activity. The same was true for Nalpha-D-valyl-lysine-vasopressin. None of the latter three analogues showed any inhibitor properties to the action of lysine-vasopressin. The two hormonogens (triglycyl N-terminal extensions) had to be given in doses 10 times greater to obtain a vasoconstrictor effect in gut and uterus equivalent in amplitude to that of a lysine-vasopressin, but this effect was still present to the same degree 2 h later with the hormonogen of lysine-vasopressin, and was only starting to return to baseline values at the same time with the arginine-vasopressin hormonogen. The vascular potency of both mono-carba L-analogues was higher than that of lysine-vasopressin, and the effect was as prolonged as with the hormonogens. The dicarba analogue also showed a prolonged action, but with much reduced potency. No significant changes in renal or myocardial blood flows were observed at all. Molecular features of vasopressin smooth muscle activity were discussed, and a receptor model was proposed. It was suggested that the -S-S-, -CH2CH2-bridges in the above analogues are not directly bound in the peptide-receptor complex and constitute the limiting factor determining complex duration, or persistence of the active peptide in the "receptor compartment". These results provide an experimental basis for possible clinical application of triglycyl-vasopressin and carba-vasopressin in bleeding from both gut and uterus and for induction of menstruation.


Quantitative iodination of human blood polymorphonuclear leukocytes. It was shown by Pincus and Klebanoff that a correlation existed between leukocytic iodination measured in vivo and microbicidal leukocytic activity. We have analyzed the results of this test in relation to time and in the presence of variable quantities of polymorphonuclear leukocytes (PMN). The values observed per time and PMN unit proved to be equivalent in the presence of 2.5 X 105 PMN or 5.0 x 105 PMN per 0.5 ml of incubation medium, measured after 10, 20 and 30 minutes or in the presence of 1.0 x 106 PMN, measured after 10 minutes. That is to say iodination is proportional to leukocyte concentration and incubation time. Increase of either the quantity of cells or the incubation time, beyond the area we defined, reduced iodination per cell and per unit of time. Concerning the patients with an insufficient iodination, we have studied 2 parameters in the presence of 5.0 x 105 PMN: 1) initial iodination measured after 10 and 20 minutes and 2) stability of iodination measured after 60 minutes. These two parameters were equally affected in two cases with myelofi-rosis, 3 patients with acquired refractory anaemia, one with chronic lymphoid leukaemia, one with erythroleukaemia, one with hairy cell leukaemia, one with systemic mastocytosis and almost complete myeloperoxidase dificiency, one with sickle cell disease, two with liver diseases and two with chronic myeloid leukaemia. The iodination at the 60th minute was more affected than at the 10th minute with a patient with myelofibrosis and 4 other patients with acquired refractory anaemias. The significance of these differences is not well understood; however the meaning of the decrease in the iodination of whatever type is that a PMN anomaly exists directly related to the myeloperoxidase H2O2 halogenation system, or to one of the stages of engulfment and/or metabolic events preceeding it and leading to the production of H2O2. This test, with the alterations we introduced, is suggested as a test for detection of functional PMN abnormalities.


Binding of phenoxybenzamine to rat platelets and platelet fractions. Irreversible binding of labelled phenoxybenzamine to platelets suspended in Tyrode solution was complete by 30 min and was proportional to the concentration incubated and non-saturable. Evidence for a reversible interaction was obtained at earlier time periods. Firmly bound phenoxybenzamine was found mainly in the supernatant (105,000 times g) of platelet homogenates. Small amounts were also detected in the 10,000 times g pellet. The labelled material in the supernatant fraction appeared to have a molecular weight of 500,000 as indicated by gel column chromatography. It was non-dialyzable, precipitated by 40-80% ammonium sulfate, and was absent in smooth muscle from the vas deferens. The material was found to bind norephinephrine but did not alter platelet aggregation. A platelet thrombosthenin fraction but not albumin or platelet fibrinogen bound phenoxybenzamine and had properties similar to the material in the supernatant fraction. The relationship of the phenoxybenzamine-binding material to thrombosthenin is discussed.


Papaverine, cyclic AMP and the dependence of the rat aorta on extracellular calcium. The spasmolytic effects of papaverine and dibutyryl cyclic AMP (db-cAMP) were compared on isometric contractile responses induced by addition of increasing amounts of external calcium to K+-depolarized or noradrenaline-stimulated rat aorta strips. Papaverine at a concentration active on depolarized strips (3 times 10(-5) moles/1) reduced the maximal contraction (Emax) elicited by Ca2+ in these preparations, while db-cAMP did not. Contrary to what was observed on depolarized aortae, the degree of inhibition of noradrenaline-stimulated strips did not decrease with increasing extracellular calcium concentration (Ca)e. Both db-cAMP and papaverine at a concentration which did not depress Emax (5 times 10(-6) moles/1) potentiated the relaxing effect of high (Ca)e on contractions elicited by noradrenaline. In conclusion, cyclic AMP is probably implicated in the mode of action of papaverine on the noradrenaline-stimulated rat aorta. At a concentration active on depolarized strips, papaverine is also able to impair contractility directly.


Central and peripheral contribution to the antihypertensive action of indoramin. Indoramin has been reported to reduce blood pressure in experimental animals and humans. The complex pharmacological profile of the compound suggests that it may also exert central actions. Effects on spontaneous sympathetic outflow and on sympathetic nerve and blood pressure responses to hypothalamic stimulation were examined in anesthetized cats in the present study. At a low dose (1 mg/kg) indoramin reduced blood pressure but did not significantly influence the level of efferent sympathetic nerve activity. A higher dose (5 mg/kg) lowered pressure further and inhibited activity in the splanchnic, cardiac and renal nerves indicating that reduced sympathetic outflow is a contributing factor to the hypotensive response at this dose. However, in contrast to other centrally acting antihypertensive agents, e.g., clonidine, indoramin did not inhibit the increase in sympathetic nerve activity produced by hypothalamic stimulation.


Influence of atropine, metiamide and vagotomy on cAMP of resting and stimulated gastric mucosa. In normal rats the effect of atropine and metiamide was studied on cAMP levels of resting and histamine-, pentagastrin-, carbachol- and insulin-stimulated gastric mucosa. In vagotomized rats gastric mucosal cAMP levels were investigated under basal conditions and after insulin. Atropine and metiamide did not alter cAMP levels of resting gastric mucosa. Truncal vagotomy caused an increase in gastric mucosal cAMP, which was not affected by insulin. All gastric secretagogues caused a significant rise in gastric mucosal cAMP levels, which was not antagonized by atropine. Metiamide effectively blocked the rise in cAMP concentration after histamine and pentagastrin, but not that evoked by cholinergic stimulation. The results are consistent with the view that the in vivo rise of gastric mucosal cAMP after carbachol or insulin is not due to a direct cholinergic action. From the spectrum of inhibitory actions of metiamide it looks as if the increase in rat gastric mucosal cAMP concentration after histamine and pentagastrin administration is mediated by H2-receptor stimulation.


Regional cyclic GMP content in incubated tissue slices of rat brain. Incubated tissue slices from different regions of the rat brain contained cyclic guanosine 3',5'-monophosphate (cyclic GMP) in the following descending order of content: cerebellum greater than hypothalamus greater than striatum greater than thalamusmidbrain greater than brain stem greater that hippocampus greater than cerebral cortex. Cholinomimetic agents at 10(-5) M (carbamylcholine, pilocarpine, acetylcholine and eserine) and papavering at 14(-4) M significantly elevated cyclic GMP accumulation in the cerebral cortex. Three min incubation with carbamylcholine (10(-5) M) did not increase cyclic nucleotide accumulation in the remaining brain regions.


The pharmacology of the inhibition of dorsal horn neurones by impulses in myelinated cutaneous afferents in the cat. The effects of microelectrophoretic strychnine and bicuculline methochloride were studied on the time course of synaptic inhibitions of single dorsal horn neurones in the lumbar spinal cord of cats anaesthetized with pentobarbitone. The inhibitions, evoked by volleys in mixed myelinated cutaneous afferents, varied in latency and duration. In general, early inhibitions (latency less than 12msec; duration less than 36 msec) were reduced by microelectrophoretic strychnine whereas late inhibitions (latency more than 16 msec and more prolonged in duration) were usually sensitive to bicuculline. These results can be interpreted in terms of glycine and GABA as the inh  ibitory transmitters of early and late inhibitions respectively.


Effect of hypoxia on the cerebral energy state. The effect of hypoxia on glycogen content, adenyl cyclase activity, cyclic 3',5'-AMP concentration, lactate: pyruvate ratio and the energy charge potential connected with the pool of adenylates was evaluated in the motor area of the cortex of beagle dogs. At first, controlled hypoxia induces activation of the adenyl cyclase system while subsequently, with PaO2 values lower than 20 mm Hg, a depression of the system is observed simultaneously with a drop of the energy charge potential. When normal oxygenation is resumed, the biochemical parameters examined tend to go back to normal even though it is still necessary to resort to anaerobic mechanisms. The intracarotid perfusion with nicergoline enhances the post-hypoxic recovery of adenyl cyclase activity, even though the ergoline derivative is inactive during the stages of hypoxic suffering of the brain studied in the present work.


Immediate effect of prostaglandin F2alpha during the luteal phase of the menstrual cycle. Serum luteinizing hormone, estradiol, 17-hydroxyprogesterone, and progesterone determinations were performed in five normally ovulating women prior to, during, and after prostaglandin F2alpha (PGF2alpha was infusion. A total of 75 mg of PGF2alpha was infused over 48 hours, beginning on the 4th day of the luteal phase. Endometrial biopsies were obtained at the completion of the infusion. In all patients, transient decreases in serum progesterone and 17-hydroxyprogesterone levels were observed during PGF2alpha infusion. Subsequent rises in the serum progesterone and 17-hydroxyprogesterone levels were observed 24 hours after completion of the infusion. There was no significant change in serum luteinizing hormone levels. Premature menstrual bleeding occurred with 24 to 72 hours after the termination of the infusion, while progesterone and 17-hydroxyprogesterone levels were still elevated. Endometrial histology revealed focal stromal necrosis in the presence of the normal secretory endometrium. It is concluded that, in the human, PGF2alpha at this dose and time schedule is not luteolytic.


[Effect of calcium on steroidogenesis in isolated human adrenal cells (author's transl)]. Isolated adrenal cells were obtained surgically from patients with primary aldosteronism, breast cancer, or Cushing's syndrome. They were prepared by the modification of Sayers method, and incubated at 37 degrees C for 2 hours under 95% O-2-5% CO-2, in the medium of calcium-free Krebs-Ringer bicarbonate buffer containing 0.2% glucose and 0.5% bovine serum albumin, to which various doses of calcium, ACTH, dibutyryl cyclic AMP or cycloheximide were added. Steroid production was measured by the method of Silber et al. In isolated normal adrenocortical cells, 11-OHCS was produced by calcium alone in the absence of ACTH or dibutyryl cyclic AMP, while it was not produced by ACTH alone without calcium. 11-OHCS production by ACTH was decreased in the high concentration of calcium (10.16 mM, 12.70 mM). Cycloheximide partially blocked an increase in 11-OHCS synthesis induced by calcium. These data suggest that adenyl cyclase of human adrenocortical cells may be stimulated by calcium alone, supporting the notion that calcium is a second messenger. The ratio of 11-OHCS production by calcium alone to that by dibutyryl cyclic AMP was higher in adenoma cells than in normal cells. This may account for the character of autonomic steroid production in adrenocortical adenoma cells.


Calcium requirement for stimulation of cyclic AMP accumulation in anterior pituitary gland by LH-RH. Removal of Ca2+ from the incubation medium by addition of 2 mM ethylene glycol bis-(beta-aminoethyl ether)-N, N'tetraacetic acid (EGTA) leads to at least 75% inhibition of the luteinizing hormone-releasing hormone (LH-RH)-induced accululation of adenosine 3'5'-monophoshpate (cyclic AMP) in rat anterior pituitary gland in vitro. This inhibitory effect of EGTA is reversed by the addition of Ca2+. A half-maximal effect of Ca2+ on LH-RH--induced cyclic AMP accumulation is observed at 2-5 X 10-5 M free Ca2+. The LH-RH-induced LH and FSH release is completely dependent upon the presence of Ca2+ in the incubation medium, a half-maximal effect being measured at 1-2 X 10-4 M free Ca2+. The basal release release of LH is increased upon Ca2+ removal.


Testicular androgen-binding protein (ABP):  comparison of ABP in rabbit testis and epididymis with a similar androgen-binding protein (TeBG) in rabbit serum. Testicular androgen-binding proteins (ABP) in rabbit testis, caput epididymis and efferent duct fluid (EDF) were compared to a similar androgen-binding protein TeBg) in rabbit serum. The affinity of these proteins for 5alpha-dihydrotesterone (DHT) at 0 degrees C (KaABP = 1.6 X 10(9) M-1 and KaTeBG = 1.9 X 10(9) M-1) and their steroid specificities were similar (DHT greater than androstanediol greater than progesterone and androstenedione). ABP and TeBG had also almost identical Stokes radii (42.8 +/- 1.2 and 43.9 +- 0.8 A, respectively), sedimentation coefficients (4.7 +/- 0.2 S and 4.4 +/- 0.2 S, respectively) and electrophoretic mobility (Rf = 0.4 in 6 1/2% polyacrylamide gels). Calculation of molecular weights from Stokes radii and sedimentation rates indicated a molecular weight of 74,000 (69,000-78,000) for TeBG and 76,000 (71,000-82,000) for ABP. The corresponding frictional ratios were 1.61 for TeBG and 1.55 for ABP assuming a partial specific volume (v) of 0.70 cm3/g. Polyacrylamide gel electrophoresis (PAGE) at different gel concentrations gave a mean molecular radius of 2.74 nm, also indicating  a molecular weight of about 75,000 (v = 0.70 cm3/g. ABP and TeBG could not be separated by PAGE; however, partial separation of ABP and TeBG was achieved by isoelectric focusing and ion-exchange chromatography on DEAE-cellulose. TeBG focused at pH 5.4, whereas ABP formed a distinct peak of bound radioactivity at pH 4.7. Also by ionexchange chromatography, ABP in both testis and epididymal supernatants was shown to have an apparently higher surface charge than TeBG in rabbit serum. The concentration of ABP in efferent duct fluid (2 X 10(-7) M = 60 pmol/mg protien) was much higher than TeBG in male rabbit serum (5.2 X 10(-8) M = 0.7 pmol/mg protein). These findings ruled against the possibility that ABP in the testis and epididymis could have been derived directly from serum. It is concluded that ABP and TeBG are very similar if not identical proteins both serving as transport and carrier proteins in their respective compartments.


The metabolism of luteinizing hormone. Plasma clearance, urinary excretion, and tissue uptake. The kinetics of plasma clearance, tissue uptake, and urinary excretion of tritiated ovine pituitary luteinizing hormone in adult male rats are reported. Most of the intravenously injected tritiated gonadotropin is cleared from circulation with a half-life of five minutes, and this is independent of the injected amount of hormone over a wide dose range. It was found that the hormone is rapidly removed from circulation by the kidneys, probably by glomerular filtration, and excreted in the urine. The radioactivity present in the urine is associated with material of the same molecular size as the native hormone and, moreover, the urinary hormone retains a significant amount of biological activity. A small amount of the hormone is catabolized by the kidney and liver, and our data suggest that this occurs in the cortex and hepatocytes, respectively.


Effects of follicle-stimulating hormone on cultures of Sertoli cell preparations. Sertoli cell-enriched preparations were obtained by sequential enzyme treatment of testes of 18-20 day old rats, and were maintained in culture in a chemically defined medium. The addition of highly purified follicle-stimulating hormone (FSH) to cells immediately after preparation, or after 48 h in culture, elicited an increase in the level of cyclic adenosine 3',5'-monophosphate (cAMP) when incubated in the presence of 3-isobutyl-1-methylxanthine (MIX). Luteinizing hormone (LH) had no effect on the cAMP levels. The cells cultured in the presence of FSH or dibutyryl cAMP for 48 h incorporated more [3H]leucine into trichloroacetic acid (TCA)-insoluble material than did cells cultured in the basal medium. Cycloheximide abolished the amino acid incorporation into protein precipitated by TCA. The data demonstrate that the Sertoli cell is a target cell for FSH action, and indicate that added dibutyryl cAMP can duplicate the enhancement of amino acid incorporation into protein elicited by FSH.


The excretion of proinsulin and insulin in urine. Proinsulin-like components (PLC) and insulin have been measured in 24 hr urine samples from 8 healthy subjects. The mean excretion of PLC was 45.8 ng and that of insulin 314 ng; the PLC: insulin ratio was 0.14. Urinary PLC was increased 3.5 fold in a patient with a pancreatic islet cell tumor and the PLC: insulin ratio was 0.35. The urinary PLC: insulin ratio is lower than that of serum, presumably because of the relatively lower urinary clearance of the larger molecular weight PLC.


The effect of phenformin upon the plasma pancreatic and gut glucagon-like immunoreactivity in diabetics. Five patients with mild maturity-onset diabetes were given 250 ml of a 20% glucose solution by intraduodenal infusion and eight other patients similarly received an amino acid solution in a dose of 0.5 g amino acids per kg body weight. The pancreatic and gut glucagon-like immunoreactivity (pancreatic GLI and gut GLI) in plasma were measured before and after the application of the two stimuli. Each person was tested twice; the first (control) test was followed by a second test after three days of treatment with phenformin 150 mg daily, plus the same 150 mg dose taken 60 min before the intubation. The plasma pancreatic GLI increased slightly during both infusions, but was not affected by phenformin. Intraduodenal infusion of both glucose and the amino acid solution induced a greater rise in plasma gut GLI. After treatment with phenformin, the fasting plasma gut GLI was higher than the control value in eleven of thirteen patients. In most cases higher gut GLI plasma levels were also found after duodenal administration of glucose and amino acids. These data furnish further evidence of the local action of antidiabetic biguanides on the intestinal wall, including its hormonal activity. The hypothesis is advanced that the phenformin-induced increase in gut GLI secretion may bring about competition of the latter with pancreatic glucagon for receptors in liver cell membranes, reducing the effect of glucagon on the liver, and thus contributing to a decrease in glycaemia.


Diurnal and seasonal variation in oral glucose tolerance: studies in the Antarctic. Oral glucose tolerance tests were performed in 12 young men in the Antarctic. Tests were done in the morning and afternoon at three monthly intervals (March, June, September, December). The characteristic diurnal variation in glucose tolerance persisted throughout the year, but was greater in the warmer months (March and December). There were significant seasonal differences in glucose tolerance; blood glucose values were lowest, both morning and afternoon, in December (midsummer). In one pair of tests performed during a reversal of normal sleep/activity pattern, the direction of the normal diurnal variation of glucose tolerance was also reversed.


Effect of hypophysectomy on protein synthetic-activity of free and bound hepatic ribosomes from insulin-deficient rats. Acute insulin deficiency in rats results in a decrease in the in vitro protein synthetic activity of isolated hepatic membrane-bound ribosomes and an increase in activity of free ribosomes. These changes are prevented by concomitant insulin treatment and are reversed by the administration of insulin. The current study evaluated the role of the pituitary in the genesis of these changes. The severity of diabetes produced by streptozotocin was less in hypophysectomized (Hx) rats, and in Hx rats receiving hormone replacement, as compared with similarly streptozotocin-treated intact rats. Although acute insulin deficiency in intact rats produced the previously described increase in protein synthetic activity of free hepatic ribosomes and decrease in activity of hepatic bound ribosomes, these changes did not occur in Hx rats, even when Hx rats received replacement doses of thyroxine, ACTH, and growth hormone. Thus, the changes in hepatic protein synthetic activity that occur in rats with acute experimental diabetes mellitus are secondary to the metabolic sequalae of insulin lack and the response of the pituitary gland to insulin deficiency.


A radioreceptor-assay for glucagon: binding of enteroglucagon to liver plasma membranes. A radioreceptor-assay for glucagon was developed employing pig liver plasma membranes isolated by means of an aqueous two-phase polymer system. The assay is simple, precise, and has a detection limit of 40 pg/ml. Acid-ethanol extracts of porcine intestinal mucosa and entero-glucagon purified by affinity chromatography interfered strongly with the binding of 125I-labelled glucagon. The affinity of enteroglucagon for the membranes was lower than that of glucagon, but even physiological concentrations of the former interfere with glucagon binding, indicating that enteroglucagon may compete with pancreatic glucagon for binding to the hepatocyte under physiological conditions.


Factors governing glucose induced elevation of cyclic 3'5' AMP levels in pancreatic islets. Adenosine 3'5'-cyclic monophosphate (cAMP) levels of isolated perifused pancreatic islets were elevated by high levels of glucose concomitantly with initiation of enhanced insulin secretion. The rise of cAMP was biphasic and seemed to be related to the temporal biphasic kinetics of insulin release. However, the temporal profiles of cAMP level changes and of insulin release differed; the major rise of the cAMP levels was seen during the initial phase, whereas insulin secretion was more pronounced during the second phase of release. Glucose-induced cAMP elevation required the presence of extracellular Ca++. Mannoheptulose completely blocked cAMP elevation due to high glucose. Exogenous insulin which has been shown by others to inhibit insulin secretion in vitro, blunted the glucose-induced cAMP rise. These observations and data in the literature are compatible with the concept that under physiological conditions glucose governs the intracellular cAMP levels in a Ca++ dependent manner - either directly or indirectly through metabolic effects.


Relationship between Epstein-Barr virus-associated nuclear antigen and cell viability in human cell lines. The relationship between the cell viability and frequency of cell with Epstein-Barr virus (EBV)-associated nuclear antigen was investigated in cultures of 4 EBV-carrier lymphoblastoid cell lines, by means of anti-complement immunofluorescence. The percentages of viable cells and those of EB nuclear antigen-positive cells were close to each other in the entire course of cultivation of all cell lines. This leads to a conclusion that absence of EB nuclear antigen in a given cell population should be evaluated in relation to cell viability.


A tumor-specific cytotoxic and neutralizing factor in rats immunized with ascites hepatoma induced by azo dyes. Transplantation immunity of Donryu rats against ascites hepatoma AH-64A induced by azo dye was demonstrated by intraperitoneal injection of tumor cells pretreated with heteroantibodies in vitro. Hyper-immunity was induced by successive challenges with fresh tumor cells. The cytotoxic effect of the serum of resistant rats (RRS) against AH-64A tumor cells was not reduced after absorption with normal rat liver cells, but was slightly reduced after absorption with normal rat spleen cells. The cytotoxicity was absorbed completely with 5 times 10(6) AH-64A tumor cells. AH-64A, -B, -C, and -D are ascites hepatoma cell lines originating from a single Donryu rat. AH-64A and AH-64B cross-reacted with RRS while AH-64C and AH-64D, chemically induced DBLA-6 leukemia cells and normal lymph node cells of rats, did not react with RRS in indirect immunofluorescence and cytotoxicity tests. A neutralization test was carried out by treating 2 times 10(5) tumor cell with either RRS or immune spleen cell in vitro and then injecting them subcutaneously into irradiated rats (400 R). It was found that 1:20 dilution of RRS protected the rats against AH-64A tumor cell growth while 1:40 and 1:80 dilutions of RRS caused some protection. A subcutaneous tumor mass developed after transplantation of tumor cells treated with RRS, but after about 2 weeks this began to decrease in size and disappeared completely within 6 weeks after transplantation. Treatment of AH-64A tumor cells with immune spleen cells at cell-to-cell ratios of 1:200 and 1:100 caused complete neutralization while normal spleen cells at a ratio of 1:200 had slight effect. Treat;ent with immune spleen cells prevented tumor growth from t;e start. Most of the surviving animals were resistant to c,allenge with 1 times 10(5) fresh AH-64A cells. RRS was fractionated by cellulose acetate membrane electrophoresis and the amounts of beta1- and gamma-globulin fractions were found to be 48 and 42% more than in normal rat serum. The immunoelectrophoretic pattern of resistant rat serum showed a stronger IgM precipitin line than that of normal rat serum.


Antitumor activity of amino acid derivatives in the primary screening. Approximately 350 amino acid derivatives were synthesized and tested for antitumor activity in four tumor systems. The effect on life prolongation and tumor growth was examined using mouse leukemia SR-61, Ehrlich ascites carcinoma, ascites sarcoma-180, and rat ascites hepatoma (AH-60C). Among these 350 derivatives, 29 compounds were found to be significantly effective in prolongation of the median life-span and inhibitory effect on tumor growth in the primary screening. Among these 29 compounds, the following five compounds were found to possess potential antitumor activity: N-(2-Naphthalene)sulfonyl-DL-tryptophan (A-91), 2-naphthylaminomethyl-gamma-aminobutyric acid (A-144), N-ethylcarbaminomethyl-L-isoleucine (A-145), N-9-fluorenylacetyl-L-phenylalanine (A-192), and N-propionyl-L-valine (A-195). These five compounds were active in prolongation of the life-span of mice bearing Ehrlich ascites carcinoma and in the inhibition of the cell growth. Some of these amino acid derivatives inhibited biosynthesis of macromolecules, DNA, RNA, and protein, in tumor cells. These results suggest that the site of action of the five amino acid derivatives appears to result from the inhibition of macromolecules and another unknown mechanism.


Effect of steroid hormones on growth of adrenocortical carcinoma transplants in mice;. The growth of transplanted carcinomas originating in a gonadectomized C3H male mouse was much faster in intact male mice than in intact or gonadectomized females. Administration of 17beta-estradiol or testosterone propionate in pellets resulted in a marked acceleration of the carcinoma growth in intact female mice. Progesterone and deoxycorticosterone acetate exerted no influence on the carcinoma growth. Neither gonadotropins (pregnant mare's serum gonadotropin plus human chorionic gonadotropin) nor ACTH accelerated the growth. In contrast, cortisone acetate in pellets markedly reduced the growth rate of carcinoma transplants when administered simultaneously with the transplantation. Cortisone acetate also arrested the growth-accelerating effect of 17beta-estradiol on transplanted carcinomas.


Combination effect of bis(3-methylsulfonyloxypropyl)amide and 6-mercaptopurine on AH-60C and AH-109A. Survival period of rats bearing ascites hepatoma, AH-60C, which was less sensitive to bis (3-methylsulfonyloxypropyl)amine (864-T) and 6-mercaptopurine each alone, was prolonged by the combined treatment with these two agents. Two or 3 rats in groups of 12 animals each survived over 60 days by this combined therapy beginning from day 0. In AH-109A, which was insensitive to 6-mercaptopurine, the group treated with this combination did not survive longer than that with 864-T alone.


Heat sensitivity of murine sarcoma virus. Murine sarcoma virus (Moloney) was found to be heat labile. Inactivation curve represents a first-order kinetics at temperatures between 30 degrees and 60 degrees. Inactivation constant analysis showed an inflexion at about 45 degrees, and the activation energy was calculated as 67.6 kcal/mol for higher temperature range and 7.95 kcal/mol for lower temperature range. Nature of inactivation is also discussed.


Watery diarrhea associated with pancreatic islet cell carcinoma. A patient with severe watery diarrhea and a non-beta islet cell carcinoma of the pancreas producing five hormones (secretin, serotonin, enteroglucagon, vasoactive intestinal polypeptide, and pancreatic glucagon) is described. We have demonstrated massive pancreatic hypersecretion to be a major factor in this patient's diarrhea. Possible inter-relationships of the actions of five hormones present in excess in the patient are discussed.


Modulation of stimulated acid secretion by dibutyryl cyclic AMP in the dog stomach. An in vivo canine gastric chamber preparation was used to determine effects of exogenous cyclic AMP and dibutyryl cyclic AMP on histamine-driven acid secretion and the role of blood flow in these responses. Histamine was infused intravenously at a submaximal rate 1.2 mu/kg-min) followed by the intraarterial infusion of either cyclic AMP (0.15 mg/kg-min) or dibutyryl cyclic AMP (0.15 mg/kg-min. Acid secretion and mucosal blood flow (aminopyrine clearance) were measured at 15-minute intervals, and electrical potential difference was measured continuously. Histamine alone decreased the potential difference from 82 mV to 48 mV within 60 minutes, increased aminopyrine clearance from 1.0 ml/min to 2.4 ml/min and increased acid output to 220 muEq/15 min. All values returned towards control following termination of histamine infusion. Dibutyryl cyclic AMP significantly increased histamine stimulated acid secretion to 350 muEq/15 min, but did not alter mucosal blood flow or electrical potential. Cyclic AMP did not change secretory, circulatory or electrical response to histamine. Therefore, dibutyryl cyclic AMP appears to increase histamine-stimulated acid secretion, and the effect is not mediated by increased mucosal blood flow.


Comparison of metronidazole and chloroquine for the treatment of amoebic liver abscess. A controlled trial. Chloroquine and metronidazole were compared in a randomized trial for the treatment of amoebic liver abscess in 36 patients. An additional 30 patients were treated without randomization. Chloroquine was given according to the standard protocol at Los tangeles County-University of Southern California Medical Center, which is 500 mg daily for 10 weeks, whereas metronidazole was given in a dose of 750 mg three times daily for 10 days. Criteria for the diagnosis of amoebic abscess included (a) a suitable clinical picture, (b) filling defect on hepatic scan, (c) high titer of antibody to Entamoeba histolytica by indirect hemagglutination, and (d) eventual complete recovery with treatment or appropriate findings on autopsy examination. Treatment failure, defined as illness that persisted beyond 10 days or recurred after that time, occurred in 1 of 28 patients treated with chloroquine and in 2 of 36 treated with metronidazole. In 2 patients who died it was difficult to assess the result of drug therapy. We conclude that both drugs are highly effective. Metronidazole has the advantage of effectiveness against intestinal amoebiasis and the probability of cure with a shorter course of treatment. Speed of response was slightly greater with metronidazole and was somewhat related to abscess size.


A controlled evaluation of intravenous adrenocorticotropic hormone and hydrocortisone in the treatment of acute colitis. This study compares the efficacy of intravenous adrenocorticotropic hormone (ACTH) with intravenous hydrocortisone in the treatment of patients with symptomatic inflammatory bowel disease. Drug doses were pharmacologically equivalent on the basis of achieved plasma cortisol levels and continuously monitored urinary corticoid excretion rates. Drug selection and patient evaluation were accomplished with a random double blind technique. Evaluation of 22 consecutive hospital patients indicates that ACTH and hydrocortisone, when administered intravenously in pharmacologically equivalent dosage, are therapeutically equivalent, that response to ACTH is rapid, with no therapeutic lag, and that differences in therapeutic responses cannont be corrrelated with differences in systemic steroid levels.


Effects of drugs influencing gastric secretion on the quantitative histological distribution of cyclic adenosine 3':5'-monophosphate in the rat stomach. To help clarify conflicting evidence on the cyclic adenosine 3':5'-monophosphate (cyclic AMP) role in acid secretion in the rat stomach, and to establish the quantitative effects on cyclic AMP concentrations in histological regions of the glandular stomach of the 24-hr fasted rat of injections of drugs that stimulate or inhibit gastric secretion, the concentrations were measured by the firefly luminescence method using fresh-frozen microtome tissue sections cut serially through the stomach wall. The data are compatible with cyclic AMP involvement in the acid secretion. Parietal-mucous neck cell cyclic AMP levels (3.3,0.11 means, expressed as picomoles per mg of wet weight and picomoles per mug of protein-nitrogen, respectively) were the highest in any region. Pentagastrin or urecholine elevated these values slightly, and theophylline produced greater increases in all regions (parietal-mucous neck cell, 5.4, 0.22), but theophylline administered either with pentagastrin or urecholine further elevated the values (8.3, 0.31). Urecholine or pentagastrin plus theophylline each generated two approximately equal cyclic AMP concentration peaks (about one-half parietalmucous neck cell concentrations (2.7, 0.22), perhaps by negative feedback, but low dose (2 mg per kg) elevated these values more than pentagastrin or urecholine, and with theophylline they were greatly increased (9.1, 0.32). The inhibitors, atropine and prostaglandin E2, alone or with theophylline, had little influence on parietal-mucous neck cell cyclic AMP levels but caused an increase in chief cell and muscle zones.


Effects of p-chloromercuribenzene sulfonate upon ultrastructure of canine gastric surface cells. Exposure of the mucosal surface of exteriorized chambered fundic segments of dog stomach to p-chloromercuribenzene sulfonate (PCMBS) resulted in predictable gross and microscopic structural changes. The first visible response after PCMBS (10 mM) in isosmotic HCl was an intense release of periodic acid-Schiff-stainable mucus. Within 2 min, radioactive PCMBS was found within cells along the entire depth of the gastric glands. Surface cells at 15 min revealed vacuolization of the cell cytoplasm, dilation of the perinuclear and intercellular spaces, mitochondrial swelling, loss of microvilli, and plasmalemma irregularities. Tight junctions, Golgi, and mucous granules appeared unaltered. However, in advanced injury, mucous granules became less dense and their membranes, which were usually smooth, became fragmented and convoluted at this point. Necrotic parietal cells were extruded into the lumen. Damage, however, only rarely extended into the deeper glandular and connective tissue areas.


Effects of cyclic adenosine 3':5'-monophosphate and related agents on acid secretion by isolated rabbit gastric mucosa. The effects of various adenosine phosphate compounds, theophylline, histamine,a nd metiamide, on steady rates of acid secretion by isolated fundic mucosa of the rabbit were measured by the pH stat method. Cyclic adenosine 3':5'-monophosphate (cyclic AMP), N(6), O(2')-dibutyryl adenosine 3':5'-monophosphate and theophylline increased the rate of acid secretion. Addition of theophylline in a concentration which has no stimulatory effect, reduces the effective concentrations cyclic AMP or histamine required for stimulation of acid secretion. Measurements of lactate, pyruvate, and CO2 appearances indicated that the increases in acid secretory rates were predominantly due to H+ and not organic acid accumulation in the luminal bath-secretion. Metiamide prevented the stimulatory effects of histamine and ATP. However, metiamide did not prevent the stimulatory effects of N(6),9(2')-dibutyryl adenosine 3':5'-monophosphate, theophylline, or 5'-AMP. The results provide further evidence for a role of cyclic AMP in governing the rate of acid secretion by rabbit stomach. The data also are consistent with histamine and ATP (at least in the concentration used) requiring adenylate cyclase activity for stimulation of acid secretion and 5'-AMP inhibiting phosphodiesterase activity.


Epidemiology of herpesvirus and respiratory virus infections. Part 1. Serologic findings. The majority of the population group studied had complement-fixation antibodies toward the following viruses: influenza type A, respiratory syncytial, cytomegalovirus, and Epstein-Barr. The herpesvirus infections (cytomegalovirus and Epstein-Barr virus) seemed to be prevalent. Only low incidences of antibodies were found toward adenovirus, influenza type B, influenza type C, and parainfluenza type II. A total of 70 aucte virus infections (increases of antibody titer) were diagnosed in 49 patients. Besides cytomegalovirus, no particular virus infection occurred in a large number. Only 11 of the 70 acute virus infections diagnosed serologically were accompanied by clinical signs of disease


ACTH-secreting 'apudoma' of gallbladder. The case of a 44-year-old woman is reported. The diagnosis after the appropriate tests and laparotomy was ACTH-secreting 'apudoma' of the gallbladder. This is a rare tumour and this case is believed to be the first reported of an ectopic hormone producing tumour from this side.


[Sensitivity to noradrenaline and cold tolerance of rats treated chronically with ACTH and adrenocortical hormone (author's transl)]. Rats treated with ACTH and hydrocortisone for 4 weeks were infused i.v. with noradrenaline or exposed to cold at -15 degrees C for 2 hours, and changes in the rectal temperature and plasma concentrations of NEFA, sugar, and lactate were observed. Rats received single injection of ACTH and hydrocortisone, warm- and cold-acclimated rats were used as controls. In ACTH- and hydrocortisone-treated rats, the noradrenaline infusion caused a significant elevation of the rectal temperature, but lowering of the rectal temperature induced by the acute cold exposure in these treated rats did not differ from the changes in control groups. Moreover, considerable differences were found between these treated rats and cold-acclimated ones in plasma concentrations of NEFA, sugar and lactate after noradrenaline infusion and acute cold exposure. From the results it was inferred that chronic treatment with ACTH and hydrocortisone did not produce similar adaptive metabolic alterations to those found in the course of cold acclimation.


Somatostatin-induced inhibition of insulin secretion from isolated islets of rat pancreas in presence of glucagon. In order to study the oeffect of somatostatin on the endocrine pancreas directly, islets isolated from rat pancreas by collagenase were incubated for 2 hrs 1) at 50 and 200 mg/100 ml glucose in the absence and presence of somatostatin (1, 10 and 100 mg/ml) and2) at 200 mg/100 ml glucose together with glucagon (5 mug/ml), with or without somatostatin (100 ng/ml). Immunologically measurable insulin was determined in the incubation media at 0, 1 and 2 hrs. Insulin release was not statistically affected by any concentration stomatostatin. On the other hand, somatostatin exerted a significant inhibitory action on glucagon-potentiated insulin secretion (mean +/- SEM, mu1/2 hrs/10 islets: glucose and glucagon: 1253 +/- 92; glucose, glucagon and somatostatin: 786 +/- 76). The insulin output in th epresence of glucose, glucagon and somatostatin was also significantly smaller than in thepresence of glucose alone (1104 +/- 126) or of glucose and somatostatin (1061 +/- 122). The failure of somatostatin to affect glucose-stimulated release of insulin from isolated islets contrasts its inhibitory action on insulin secretion as observed in the isolated perfused pancreas and in vivo. This discrepancy might be ascribed to the isolation procedure using collagenase. However, somatostatin inhibited glucagon-potentiated insulin secretion in isolated islets which resulted in even lower insulin levels than obtained in the parallel experiments without glucagon. It is concluded that the hormone of the alpha cells, or the cyclic AMP system, might play a part in the machanism of somatostatin-induced inhibition of insulin release from the beta-cell.


Studies of the biological activity of insulin, cyclic nucleotides and concanavalin A in the isolated fat cell. The insulin or proinsulin response of the isolated rat adipocyte was destroyed by preincubation with trypsin. After 120 minutes, biological responsiveness partially regenerated. Similarly, the biological responsiveness of the isolated fat cell to non-suppressible insulin-like activity (NSILA) was only partially destroyed following trypsin digestion, and did not regenerate. In contrast to the above, cyclic AMP or dibutyryl cyclic AMP effects were unaltered by trypsin or neuraminidase digestion.


Effects of glucagon, phosphodiesterase inhibitors, and trypsin treatment on cyclic 3',5' adenosine monophosphate levels in isolated hepatocytes. Cyclic 3',5' adenosine monophosphate (cyclic AMP) levels were measured in isolated hepatocytes under several conditions. Following the addition of glucagon cyclic AMP levels increased rapidly with peak values occurring at three minutes. The increase in cyclic AMP was dose dependent. Significant increases were found with 10(-10)M glucagon and a maximum increase of twenty fold was produced by 10(-8) M glucagon. This action of glucagon was augmented by the phosphodiesterase inhibitors, theophylline, SQ 20,009, and papaverine. Treatment of the hepatocytes with trypsin markedly reduced the response to glucagon.


Irreversible inactivation of hypoxanthine phosphoribosyltransferase by periodate oxidized nucleotides. Hypoxanthine phosphoribosyltransferase (EC 2.4.2.8) from rat brain or human erytherocytes can be irreversibly inactivated by incubation with periodate-oxidized analogues of the enzyme products GMP or IMP. This inhibition is specific and directed against the product binding site of the enzyme. Inactivation is not produced by periodate-oxidized AMP or other aldehydes, for example periodate-oxidized glycerol. The inactivation is concomitant with the binding of the inhibitor to the enzyme protein. The bound inhibitor cannot be removed from the protein by dialysis, Sephadex chromatography or polyacrylamide-gel electrophoresis. Adenine phosphoribosyltransferase (EC 2.4.2.7), on the other hand, is not influenced by any of the inhibitors mentioned above.


[Increased estradiol-estrone quotient in rat liver by hydroxysteroids: an effect of the specific hydrogen transfer between steroids]. After addition of estrone to rat liver slices, a quotient of estradiol/estrone of ca. 0.1 is reached within 1 - 2 min. By additional application of 17 beta-hydroxysteroids this quotient is changed in the direction of estradiol, although the applied concentrations of both steroids are far below the concentration of the cytoplasmic redox couple NADH/NAD. Of all the steroids tested, testosterone had the strongest influence on the quotient, especially in the liver of female rats. This influence is smaller in the livers of male rats and infantile animals. The changing of the E2/E1 quotient by testosterone can be inhibited by the antiandrogen cyproteron acetate. Steroids with hydroxy groups at C-3 or C-20 or high concentrations of non-steroids, which can be oxidized by NAD, change the E2/E1 quotient only minimally. The experiments demonstrate that in liver, the redox couple estradiol/estrone is not in equilibrium with the main redox couple of the cytoplasmic NADH/NAD. Only on account of this fact it is possible that relatively low concentrations of testosterone change the E2/E1 quotient via the C-17 leads to C-17 hydrogen transfer between steroids. Biological consequences are discussed.


Adopting new models for continuity of care: the ward as mini-mental-health-center. Several years ago the staff of a traditional psychiatric ward in a Veterans Administration hospital began trying to provide continual care for their former inpatients after they left the hospital. Nursing assistants conducted basic-skills groups and other groups in board-and-care homes; staff members made themselves available to former inpatients and theri families, roommates, or board-and-care sponsors through a 24-hour telephone service and home visits; and a day-care program and follow-up groups for the ward's outpatients were set up. Later the staff establiched additional day-care and follow-up programs in a downtown satellite unit. In a recent three month period, patients made a total of 4488 visits to the satellite and hospital programs. The preceding paper discusses some of the theoretical considerations on which this program and other alternatives to traditional care are based.


On the distribution of some genetic markers in libya. On a sample of N equals 169 nonrelated blood donors from Libya several blood group, serum protein group and enzyme frequencies are within the range of those of other Caucasoid populations. However, the frequencies of cDe, Jk-a, Fy-c, Hp-1, Gc-1, aP-b and AK-1 show some greater differences and seem to indicate certain Negroid influence on the genetic composition of our Libyan sample. This assumption is conceivable considering the racial history of Libya.


Lipoproteins in lecithin-cholesterol-acyltransferase(LCAT)-deficiency. II. Further studies on the abnormal high-density-lipoproteins. The lipoproteins from two sibs with familial lecithin-cholesterol-acyltransferase(LCAT)-deficiency were further characterized.  Comparatively lipoproteins from patients with secondary LCAT-deficiency were studied.  Both groups of patients had particles of unusual size and shape in the alpha1-(HD-2)-lipoprotein subfraction.  The abnormal HDL-2 particles were disk-like in appearance with a major axis of about 180 A and a minor axis of about 40 A and tended to aggregate into long coinlike stacks.  The abnormal HDL-2 particles contained the normal protein constituents of HDL Apo A-I, Apo A-II and Apo C but in addition a major polypeptide with a M.W. of 39000 not seen in significant amounts in normal high-density-lipoproteins.  This polypeptide was found identical in size, isoelectric focusing and immunochemically with an arginine-rich normal polypeptide constituent of very-low-density-lipoproteins designated apoprotein E.  Presence of this protein marker in the HDL allowed the specific immunological detection of the abnormal HDL-2 (LP-E) in plasma.  Further minor biochemical abnormalities were observed in the lipoproteins of the patients with familial LCAT-deficiency.  However, the main protein constituents of their HDL, the Apo A, Apo C and Apo E polypeptides, were found to be identical electrophoretically and by analytical isoelectric focusing with their normal counterparts.  The data suggest that the basic genetic defect in the hereditary disease leads to a deficient activity of the LCAT-enzyme and that all abnormalities in the lipoprotein spectrum are secondary.


Enzyme polymorphisms and haemoglobin variants in Greeks. Several enzyme polymorphisms and hemoglobin variants were typed in a sample of n = 219 non-related Greek blood-donors.  The following gene frequencies were observed: pa = 0.201, pb = 0.701, pc = 0.098;PGDA = 0.985, PGDc = 0.015; AK1 = 0.942, AK2 = 0.058; HbA = 0.988, HbS = 0.012.  No polymorphic variation was seen in LDH, s-MDH, PHI, or SOD.  The population genetical aspects of these results are discussed.


Hypersarcosinemia with craniostenosis-syndactylism syndrome. Hypersarcosinemia with craniostenosis-syndactylism syndrome. After a sarcosine loading the sarcosine-glycine ratios seem to be a more reliable criterion to distinguish different genotypes than the sarcosine tolerance curves.


Heterokaryons in the analysis of genes and gene regulation. Cytological and chemical analysis of heterokaryons, the immediate product of cell fusion, offer new possibilities for studying the factors responsible for genetic regulation in eukaryotic cells. In comparison with proliferating cell hybrids the heterokaryon state offers the important advantage that a heterokaryon contains two complete genomes since chromosome loss does not occur, but since segregation and recombination are absent, heterokaryons cannot be used for gene mapping in the same way as proliferating cell hybrids. However, if two cell types carrying different genetic defects are fused the analysis can be used for studies of gene complementation. The biological information obtained with heterokaryons has emphasized the role of the cytoplasm in the control of nuclear activity. When a G1 nucleus is brought into contact with the cytoplasm of an S phase cell the G1 nucleus is stimulated to synthesize DNA. If the nucleus is brought into a mitotic cell, the chromatin of the G1 nucleus is forced to condense into prematurely condensed chromosomes. Inactive nuclei such as the dormant chick erythrocyte nucleus will be stimulated to initiate RNA and DNA synthesis when brought into contact with an active cytoplasm by cell fusion. Specific nuclear proteins have been shown to be responsible for this process of reactivation. Other inactive nuclei such as the nuclei of macrophages and spermatozoa have likewise been shown to be reactivated by fusion with active cells. The degree of activation in all of these cases appears to be determined by the state of the active cell. Inactive nuclei are activated to the same level as the active nucleus but seldom beyond this level. If differentiated cells are fused with undifferentiated cells, usually the differentiated character is lost rapidly after fusion. This observation is in agreement with several studies on proliferating cell hybrids indicating some type of negative control of differentiated properties. In heterokaryons obtained by fusion of cells of a similar type of histotypic differentiation usually coexpression of the differentiated markers is observed.