Effects of diazepam on photosynthesis, respiration, rubidium uptake, and finestructure of Scenedesmus obliquus in synchronous cultures. Effects of diazepam (Valium) on photosynthesis, chlorophyll/photosynthesis ratios, respiration, uptake of rubidium ions, and ultrastructure of Scenedesmus obliquus synchronized by a light-dark regimen of 14:10 hrs were determined. 80 and 160 muM diazepam, added to the nutrient medium at the start of the light-dark change (i.e., start of the cell cycle) gradually reduced rates of photosynthesis, below the initial rates from the beginning of the experiment. Contents of chlorophyll, however, remained nearly unaffected. Consequently, the diazepam-treated cells had a higher chlorophyll/photosynthesis ratio--also with regard to respiration in order to calculate the gross photosynthesis. The occurrence of photorespiration cannot be assumed. The net influx of rubidium was slightly reduced by 100 muM diazepam 0.5 and 2.0 hrs after the start of the cell cycle and was strongly inhibited after 5 to 14 hrs. 80 and 160 muM diazepam caused separation of thylakoids, formation of giant mitochondria and enlargement of vacuoles.


Ultrastructural changes occurring during germination and outgrowth of spores of the thermophile Bacillus acidocaldarius. Spores of the thermophilic, acidophilic, Bacillus acidocaldarius were covered by a thick outer coat and a laminated inner coat (5.5 nm periodicity). Small membranous vesicles were present in the spore core and they disappeared as germination proceeded. After depolymerization of the cortex, and a 30% increase in spore diameter a localized gap appeared in the laminated inner coat only. This inner coat gap was narrow and could be the whole length of the spore. The germ cell appeared to grow, or to be pushed towards the inner coat gap, at which stage the outer coat disappeared in the same localized area. As the vegetative cell grew out the spore coat fell away, with loose cortical material still attached to it. The young germ cell developed a large spherical electron dense inclusion body in the cytoplasm, at the same time as the ribosomal and nuclear areas became distinct.


Microbial assimilation of hydrocarbons. I. The fine-structure of a hydrocarbon oxidizing Acinetobacter sp. 1. The fine-structure analysis of the hydrocarbon oxidizing microorganism, Acinetobacter sp., demonstrated a cytoplasmic modification resulting from growth on paraffinic and olefinic hydrocarbons. 2. Intracytoplasmic hydrocarbon inclusions were documented by electron microscopy with chemical identifications obtained by gas chromatography and X-ray diffraction. 3. These results demonstrate the ability of a microorganism to accumulate hydrocarbon substrates intracellularly which, in turn, indicates the transport across the cell membrane.


Duration of hospitalization for alcoholism. On admission to an inpatient alcoholic treatment center, 58 alcoholic men were randomly assigned to two groups. One group was detoxified and after-care services were planned (mean length of stay nine days). The other group, in a-dition to the above, received intensive psychosocial inhospital treatment (mean length of stay 30 days). Posthospital adjustment was measured at five intervals over a 13 month period in the areas of social relationships, financial status, employment record, legal involvement, drinking behavior, use of measures were obtained on psychological change and counseling readiness. No significant difference was found between the two groups on any measure. These results indicate that the hospital has a limited role in the treatment of the alcholic person.


Reflex patters initiated by the secondary sensory fiber endings of muscle spindles: a proposal. Muscle spindles have two kinds of sensory fibers, each with stretch-sensitive endings on the nuclear bag muscle fibers and the nuclear chain muscle fibers within the spindle.  The stretch reflex responses initiated by the primary sensory fibers have been well defines.  There are other stretch reflexes of poorly defined origin and distribution. Clinical studies on spastic patients indicate that these stretch reflexes show regional responses or regional synergies.  Stretch of flexor muscles of proximal joints of an extremity causes slowly recruiting, prolonged, reflex flexion in that extremity, in the contralateral extremity and also in the ipsilateral extremity, producing the crossed extension-flexion reflex and long spinal reflex.  Likewise, stretch of extensor muscles of the proximal joints initiates a similar extension reflex in the same distribution.  Stretch of the distal flexors of an extremity reflexly spreads flexion through that extremity.  Stretch of the distal extensors of an extremity spreads extension throughout that extremity.  These reflexes can be abolished by intramuscular neurolysis with phenol.  They have long internuncial pathways and involve both alpha and gamma motoneurons.  The demonstration that these reflex patterns are initiated by stretch of the muscle and show characteristics similar to those shown in physiological studies by the secondary sensory spindle reflexes suggests that they are initiated by stretch of the endings of the secondary sensory fibers of muscle spindles.


Pediatric cadaver kidneys. Their use in renal transplantation. Of 350 consecutive cadaver kidney transplants, 32 kidneys from donors aged 1 day to 9 years were transplanted. Our results indicate that, with strict adherence to certain guidelines in kidney procurement and transplantion, pediatric kidneys are excellent donor graft material. In contrast to en bloc transplantation of both kidneys from pediatric donors, each donor can provide kidneys for two recipients. In addition, the transplantation of pediatric kidneys as single units is both simple and safe.


Histocompatibility antigens and solid malignant neoplasms. To evaluate the relation between histocompatibility antigen phenotypes and solid malignant neoplasms, HL-A type was determined in 633 cancer patients and compared with those of 489 normal controls. HL-A8 was elevated in patients with squamous cancer, melanoma, and adenocarcinoma. The highest incidence occurred in patients with salivary gland adenocarcinoma (67% vs only 17% in normal controls). A threefold increase in HL-A5 was detected in patients with connective tissue sarcomas (28% incidence vs 9% in normal controls). Antigen frequencies did not vary when analyzed by time of diagnosis or interval after treatment. The finding that certain malignant neoplasms have associations with increased frequency of individual HL-A antigens may give clues to cause and genesis for these tumors.


Cyclic AMP response to epinephrine and shock. Hormonal regulation is fundamental to homeostasis, and the response in tissues is mediated by adenosine 3':5' equals cyclic phosphate (cyclic AMP). Since circulating levels of catecholamines are elevated during hemorrhagic shock, experiments were carried out to study the effects of epinephrine and of shock on cyclic AMP levels of plasma and of the intestine, a particularly vulnerable organ in canine hemorrhagic shock. Epinephrine was given to normotensive dogs as a constant infusion (group 1) or as a single pulse (group 2). Sequential blood samples showed a marked  increase in circulating cyclic AMP, especially in portal and hepatic venous blood. Little change was noted in the tissues. Hemorrhagic shock (group 3) induced a marked increase in plasma cyclic AMP, with a depletion of mucosal cyclic AMP and adenosine triphosphate (ATP). Pretreatment before shock with alpha-blockade (group 4) abolished the decrease in mucosal cyclic AMP and ATP, and reduced the increase in plasma cyclic AMP.


Mouse hepatitis virus-induced recurrent demyelination. A preliminary report. Four-week-old BALB/c mice inoculated intracerebrally with the JHM strain of mouse hepatitis virus developed an acute demyelinating disease followed by apparent recovery with remyelination. When surviving mice were examined 16 months later, small areas of active demyelination were still present. This is the first reported example, to our knowledge, of an experimental viral infection in which acute demyelination with recovery is followed by persisting or recurring demyelination.


Hereditary hemorrhagic telangiectasia. A case with hepatocellular carcinoma and acquired hepatocerebral degeneration. A patient with hereditary hemorrhagic telangiectasia (Osler-Weber-Rendu disease), treated with ethinyl estradiol, multiple blood transfusions, and iron-dextran, developed hepatocellular carcinoma and acquired hepatocerebral degeneration. In addition to the carcinoma, the liver contained extensive arteriovenous maliformations, telangiectasis, and changes of Osler atypical cirrhosis. The carcinoma possibly had its genesis in the presence of an ocongenic serum hepatitis virus, or the cirrhosis, or both.


Lipoprotein receptors, cholesterol metabolism, and atherosclerosis. Deposition of cholesterol esters in the arterial intima is a characteristic feature of human atherosclerosis. Very little is known about the mechanisms by which cells normally regulate their cholesterol ester content. Recent studies in cultured human cells demonstrate the existence of a cell surface receptor that binds plasma low density lipoproteins and regulates the sterol content of cells by modulating the rates of uptake, esterification, and synthesis of cholesterol. A possible role for this lipoprotein receptor in the pathogenesis of atherosclerosis is discussed.


Islet cell carcinoma of the pancreas.

Islet cell carcinoma of the pancreas. Thirty cases of islet cell carcinoma of the pancreas diagnosed at Memorial Hospital were studied. There were 17 male and 13 female patients. The average age was 44 years. Most of the tumors were located in the body or tail of pancreas; in 25 instances, the primary tumor was larger than 6 cm. Epigastric pain, hypoglycemia, and jaundice were frequent primary clinical presentations. No morphological differences were found between functioning and nonfunctioning tumors. Size of tumor, local tissue infiltration, and vascular invasion were helpful; but they were not absolute parameters aiding in the differentiation of benign and malignant tumors. Twenty-six patients had metastatic disease at time of diagnosis. Liver, regional lymph nodes, bones, and peritoneum were common sites of metastases. The average survival was 3.9 years. The cumulative five-year survival rate was 65%.


Adrenocortical activity and urinary cyclic AMP levels: effects of hypobaric hypoxia. Six highly motivated and trained military test subjects were exposed to a simulated altitude of 4267 m (477 torr) for 48 hours, prededed and followed by sea level runs lasting 32 hours. During each scenario tests subjects were required to perform their respective military tasks on a continual basis with sporadic rest not exceeding several hours. Extremely high levels of plasma cortisol prior to the start of each session and persistently elevated concentrations of urinary 17-OH corticosteroids at sea level were consistently observed. (This observation probably explains the relatively minor changes in adrenocortical output noted during acute exposure to hypobaric hypoxia. Analogous results were obtained for urinary cyclic AMP, i.e. elevated baseline levels and minimal response to high altitude. The results indicate an attenuated response as well as an association between urinary measures of adrenocortical activity and cyclic AMP excretion.


Absence of nyctohemeral variation in stress-induced ACTH secretion in the rat. Changes in plasma ACTH concentrations were measured in the adult male rat following a stressor in order to ascertain if there is a nyctohemeral variation in ACTH secretion following stress. The immuno-reactive ACTH concentrations obtained following a 2.5-min hind-leg tourniquet at 0400 or 1600 hours in pentobarbital-anesthetized animals were compared to those obtained in anesthetized and unanesthetized, unstressed controls. Neither absolute nor increment change in plasma ACTH concentrations following morning or afternoon tourniquet stress were significantly different after either 20 or 40 min anesthesia. Moreover, plasma ACTH concentrations from unstressed, unanesthetized rats showed a slight but significant difference (p smaller than 0.05) at 0400 hours [118 plus or minus 18 pg/ml (mean plus or minus SE)] compared to 1600 hours (64 plus or minus 12 pg/ml) indicating a reversal of the expected ACTH rhythm. Plasma corticosterone concentrations in these same rats showed the predicted lower-morning and higher-afternoon corticosterone pattern suggesting a possible nyctohemeral difference in the adrenal sensitivity to circulating levels of ACTH.


The use of three dye-binding procedures for the assessment of heat damage to food proteins. 1. A study has been made of pure proteins heated either alone or in contact with sugars, so as to cause a severe fall in their reactive lysine contents, and also of commercial protein concentrations. 2. For unheated materials, and for bovine plasma albumin and fat-extracted, dried chicken muscle severely heated in the absence of sugar, Acid Orange 12 binding values (mmol bound dye/kg crude protein (nitrogen times 6-25)) were close to the sum of total histidine, total arginine and reactive lysine contents (mmol/kg crude protein (N times 6-25)), which we have termed HARL values. The dye-binding values and the HARL values were reduced similarly by heat treatment. 3. For materials in which protein and glucose had reacted under mild conditions (37 degrees), the dye-binding capacity with Acid Orange 12 was unchanged even though the HARL value of these materials was considerably reduced. When protein and glucose or sucrose were heated more severely, the dye-binding capacity was slightly lowered but not to the same extent as the reduction in the basic amino acids. 4. Animal feeding-stuffs, whether unheated, industrially processed or diliberately heated, appeared to react with Acid Orange 12 in the same way as the model systems (selected to represent three types of heat damage: 'advanced' and 'early' Maillard and protein-protein damage). 5. Remazol blue binding values and fluorodinitrobenzene (RDNB)-reactive lysine values were similarly reduced in materials that had been severely heated, either with or without added sugars; however, when protein and glucose had reacted under mild conditions the fall in Remazol blue binding was less than that in FDNB-reactive lysine. 6. For the model materials, binding with cresol red was, in general, higher for heated samples but the results showed no correlation with FDNB-reactive lysine values. For meat and groundnut meals, changes in values after heat treatment were smaller than those that have been reported for soya-bean meals.


Activation of 3',5'-cyclic adenosine monophosphate phosphodiesterase by calcium ion and a protein activator. 3',5'-CAMP phosphodiesterase was partially purified from bovine cerebral cortex. A heat-stable activating factor was separated from the enzyme by chromatography on DEAE-cellulose. The enzyme in crude ammonium sulfate fractions was stimulated by 5 mM CaCl2. This stimulation was reversed by the calcium chelator EGTA. The main phosphodiesterase peak obtained by DEAE-cellulose chromatography was not stimulated by Ca2+. Upon addition of column effluent containing a heat stable factor, Ca2+ activation was restored. Protein activator was inactive when endogenous contaminating Ca2+ was complexed with EGTA. It was concluded that activation of phosphodiesterase requires the presence of both activator and Ca1+. From an analysis of activation of cGMP hydrolysis a kinetic model for the interaction of Ca2+ and protein activator with the phosphodiesterase was developed. Heterotropic cooperativity between the binding of Ca2+ and protein activator to the phosphodiesterase was observed, i.e., Ca1+ decreased the apparent dissociation constant for protein activator and protein activator decreased the apparent dissociation constant for Ca2+.


Pseudogout in ochronosis. Report of a case. Calcium pyrophosphate dihydrate crystals were identified in synovial fluid white blood cells during an episode of acute arthritis in a patient with ochronosis and chondrocalcinosis. Review of the histories and radiographs of 5 other patients with ochronosis demonstrated two additional instances of chondrocalcinosis. Both of these patients had episodes of arthritis consistent with pseudogout. This suggests that pseudogout, which has been found in increased incidence in some metabolic diseases, may also be more common in ochronosis.


Immunoglobulin deposits in skin in systemic lupus erythematosus. Immunoglobulin deposits in the dermal-epidermal junction of clinically normal skin from patients with SLE were eluted by acid buffer. The eluates contained antinuclear and antibasement membrane antibody activities. The anti-BM antibody reacted with skin and esophageal but not glomerular basement membrane. Enzymatic studies indicated that the antibody reacted with carbohydrate moeities in the basement membrane. The anti-BM antibody was not present in corresponding sera of SLE patients.


General aspects of hydrophobic chromatography. Adsorption and elution characteristics of some skeletal muscle enzymes. If the degree of substitution of Sepharose 4 B with alpha-alkylamines is varied gels of different hydrophobicity are produced. Proteins can be adsorbed when a critical hydrophobicity (ca. 10-12 alkyl residues/Sepharose sphere) is reached. The enzymes phosphorylase kinase, phosphorylase phosphatase, 3',5'-cAMP dependent protein kinase, glycogen synthetase, and phosphorylase b are successively adsorbed as the hydrophobicity of the Sepharose is increased. The capacity of the gels for these enzymes and protein in general increases exponentially reaches plateau values as a function of the degree of substitution. There is no indication of a restriction of the hydrophobic centers for a given protein. The critical hydrophobicity needed to adsorb proteins can either be otained in the above manner or by elongation of the employed alkylamine at a constant degree of substitution. Additonally, as the hydrophobicity of a gel is increased higher binding forces result and desorption of proteins requires an augmentation of the salt concentration in the elution buffer. Elution of proteins from a hydrophobic matrix can be described in terms of salting-in phenomena since desorption is dependent on the type of salt employed and not on the ionic strength alone. This also rules out ionic interactions as a major factor in adsorption per se. By rationally controlling the hydrophobicity of a Sepharose gel the adsorption and elution of a protein may be thus establised that its purification or elimination can be optimally performed.


Simple alkanethiol groups for temporary blocking of sulfhydryl groups of enzymes. New reagents for the temporary blocking of active or accessible sulfhydryl groups of enzymes have been developed. These reagents, which are either alkyl alkanethiolsulfonates or alkoxycarbonylalkyl disulfides, rapidly and quantitatively place various RS- groups on the sulfhydryls to generate mixed disulfides. In all cases native enzymes can be regenerated with either dithiothreitol or beta-mercaptoethanol. In general the temporary blocking groups also afford total protection against normally inhibitory thiol blocking agents. When RS- groups were attached to rabbit muscle creatine kinase (EC 2.7.3.2), a trend toward lower residual activities with increasing bulk was observed. Treatment of the native creatine kinase with 14CH3HgC1 led to incorporation of greater than 1 equiv of CH3Hg- group per subunit. This CH3Hg- blocked enzyme was fully active, and the blocking group afforded no protection against iodoacetamide. These results suggest that CH3Hg- and the RS- groups are modifying two different sulhydryl groups on the enzyme. When papain (EC 3.4.4.10) was treated with excess methyl methanethiolsulfonate. complete and rapid inhibition was observed, and 1 equiv of CH3S- was incorporated/mol of active enzyme. Complete protection against normally inhibitory 5,5'-dithiobis(2-nitrobenzoic acid) was afforded by the temporary blocking group. When rabbit muscle glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12) was titrated with methyl methanethiolsulfonate, two sulfhydryl groups per subunit were found to be modified, one much more rapidly than the other. If one extrapolates the initial slope of the titration curve, the inactivation of the enzyme would be complete after modification of a single cysteinyl residue per subunit.


The specificity of different classes of ethylating agents toward various sites in RNA. The alkyl products of neutral in vitro ethylation of TMV-RNA by [14C]diethyl sulfate, [14C]ethyl methanesulfonate, and [14C]ethylnitrosourea have been determined and found to differ significantly depending on the ethylating agent. Diethyl sulfate and ethyl methanesulfonate ethylate the bases of TMV-RNA in the following order: 7-ethylguanine greater than 1-ethyladenine, 3-ethylcytidine greater than 7-ethyladenine, 3-ethyladenine, O6-ethylguanosine, 3-ethylguanine. Ethyl methanesulfonate was more specific for the 7 position of guanine, and other derivatives were found in lesser amounts than with diethyl sulfate. Neither reagent caused the formation of detectable amounts (smaller than 0.26 percent) of 1-ethylguanine, 1,7-diethylguanine, N2-ethylguanine, N6-ethyladenine, N4-ethylcytidine, or 3-ethyluridine. Identified ethyl bases account for over 85% of the total radioactivity of [14C]ethyl methanesulfonate and [14C]diethyl sulfate treated TMV-RNA. Phosphate alkylation accounts for about 13 and 1%, respectively, In contrast, [14C]ethylnitrosourea-treated TMV-RNA, while reacting to a similar extent (15-70 ethyl groups/6400 nucleotides), is found to cause considerably more phosphate alkylation. Upon either U4A RNase or acid hydrolysis up to 60% of the radioactivity is found as volatile ethyl groupw in the form of [14C]ethanol, and a further 15% appears to be primarily ethyl phosphate and nucleosides with ethylated phosphate. Of the remaining radioactivity, half is found as O6-ethylguanosine, the major identified ethyl nucleoside. Other ethyl bases found in ethylnitrosourea-treated TMV-RNA are 7-ethylguanine greater than 1-ethyladenine, 3-ethyladenine, 7-ethyladenine, 3-ethylcytidine, and 3-ethylguanine. It appears that ethylnitrosourea preferentially alkylates oxygens, and that formation of phosphotriesters is by far the predominant chemical event. Since the number of ethyl groups introduced into TMV-RNA by ethylnitrosourea is similar to the number of lethal events, one may conclude that phosphate alkylation leads to loss of infectivity. None of the three ethylating agents studied are strongly mutagenic on TMV-RNA or TMV. The role of phosphate alkylation in regard to in vivo mutagenesis and oncogenesis remains to be established. At present it appears possible that the extent of this reaction may correlate better with the oncogenic effectiveness of different ethylating agents, than the extent of any base reaction. Unfractionated HeLa cell RNA is ethylated primarily in acid labile manner even by diethyl sulfate and ethyl methanesulfonate, a fact that is attributed to its high content of low molecular weight trna rich in terminal phosphates which alkylate readily.


Structural studies on carcinoembryonic antigen. Periodate oxidation. Periodate oxidation has been applied to examine the carbohydrate structure of carcinoembryonic antigen (CEA) and the possible role of the carbohydrate residues in its antigenic activity. Sialic acid (N-acetylneuraminic acid) and fucose were completely destroyed, and galactose and mannose were partially destroyed by a single periodate treatment. Serial periodate treatment (Smith degradation) destroyed additional amounts of galactose and mannose as well as significant amounts of N-acetylglucosamine. Prior removal of sialic acid by neuraminidase treatment led to increased destruction of galactose by periodate. Antigenic activity persisted indicating that the residues destroyed played little, if any, part in the antigenicity of CEA. These results yield an initial view of the structural arrangement of the carbohydrate residues in the CEA molecule.


Phase separations in phospholipd membranes. Phase diagrams representing lateral phase separations in the plane of lipid bilayer membranes have been determined for binary mixtures containing dielaidoylphosphatidylcholine together with dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dioleoylphosphatidylcholine, and dipalmitoylphosphatidylethanolamine. The phase diagrams were deduced from observations of the temperature dependence of the paramagnetic resonance spectra of low concentrations of spin-labels incorporated in these bilayer membranes. In one case, the binary mixture of dipalmitoylphosphatidylethamine and dielaidoylphosphatidylcholine, evidence has been obtained for fluid-fluid immiscibility, in specified temperature and compoistion ranges. This immiscibility could give a lateral phase separation into fluid domains in the plane of the membrane, and/or a transverse phase separation into an asymmetrical bilayer membrane, and/or possibly disco ntinuous bilayer membranes of different composition. An asymmetrical bilayer membrane can be expected on theoretical grounds to form a nonplanar membrane.


Beef-heart submitochondrial particles: a mixture of mitochondrial inner and outer membranes. 1. EPR spectra at 9 GHz and 83 degrees K of NADH-reduced anaerobic beef-heart submitochondrial particles, prepared from mitochondria by sonication and centrifugation, contain a signal (gz equals to 2.01, gy equals to 1.94, gx equals to 1.89) due to an iron-sulphur center of the mitochondrial outer membrane. 2. The ratio of inner and outer membranes in submitochondrial particles is not greatly different from that in beef-heart mitochondria. 3. Beef-heart submitochondrial particles free from outer-membrane contamination have been prepared by free-flow electrophoresis. EPR spectra at 83 degrees K of such particles are presented.


Flash photolysis electron spin resonance studies of the electron acceptor species at low temperatures in photosystem I of spinach subchloroplast particles. The light-induced electron spin resonance signals of Photosystem I spinach subchloroplast particles have been studied at approximately 6 degrees K. Using the technique of flash photolysis-electron spin resonance with actinic illumination at 647 nm, a kinetic analysis of the previously observed bound ferredoxin ESR signals was carried out. Signal I (P700+) exhibits a partial light-reversible behavior at 6 degrees K so it was expected that if the bound ferredoxin is the primary acceptor of Photosystem I, it should also exhibit a partial reversible behavior. However, none of the bound ferredoxin ESR signals showed any such light reversible behavior. A search to wider fields revealed two components which did exhibit the expected kinetic behavior. These components are very broad (about 80 G) and are centered at g equals to 1.75 and g equals to 2.07. These two components exhibit the expected characteristics of the primary electron acceptor. A model is presented to account for the reversible and irreversible photochemical changes in Photosystem I. The possible identity of the primary acceptor responsible for these two new components, is discussed in terms of the available information. The primary acceptor may be an iron-sulfur protein, but not of the type characteristic of the bound or water-soluble ferredoxins found so far in chloroplasts.


A rapid, light-induced transient in electron paramagnetic resonance signal II activated upon inhibition of photosynthetic oxygen evolution. A rapid, light-induced reversible component in Signal II is observed upon inhibition of oxygen evolution in broken spinach chloroplasts. The inhibitory treatments used include Tris washing, heat, treatment with chaotropic agents, and aging. This new Signal II component is in a 1 : 1 ratio with Signal I (P700). Its formation corresponds to a light-induced oxidation which occurs in less than 500 mus. The subsequent decay of the radical results from a reduction which occurs more rapidly as this free radical component is complete following a single 10-mus flash, and it occurs with a quantum efficiency similar to that observed for Signal I formation. Red light is more effective than far-red light in the generation of this species, and, in preilluminated chloroplasts, 3-(3,4-dichlorophenyl)-1,1-dimethylurea blocks its formation. Inhibition studies show that the decline in oxygen evolution parallels the activation of this Signal II component. These results are interpreted in terms of a model in which two pathways, one involving water, the other involving the rapid Signal II component, compete for oxidizing equivalents generated by Photosystem II. In broken chloroplasts this Signal II pathway is deactivated and water is the principal electron donor. However, upon inhibition of oxygen evolution, the Signal II pathway is activated.


The rapid component of electron paramagnetic resonance signal II: a candidate for the physiological donor to photosystem II in spinach chloroplasts. Rapid light-induced transients in EPR Signal IIf (F-+) are observed in 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU)-treated, Tris-washed chloroplasts until the state F P680 Q minus is reached. In the absence of exogenous redox mediators several flashes are required to saturate this photoinactive state. However, the Signal IIf transient is observed on only the first flash following DCMU addition if an efficient donor to Signal IIf, phenylenediamine or hydroquinone, is present. Complementary polarographic measurements show that under these conditions oxidized phenylenediamine is produced only on the first flash of a series. The DCMU inhibition of Signal IIf can be completely relieved by oxidative titration of a one-electron reductant with E'Os.o equals to + 480 mV. At high reduction potentials the decay time of Signal IIf is constant at about 300 ms, whereas in the absence of DCMU the decay time is longer and increases with increasing reduction potential. A model is proposed in which Q minus, the reduced Photosystem II primary acceptor, and D, a one-electron 480 mV donor endogenous to the chloroplast suspension, compete in the reduction of Signal IIf (F-+). At high potentials D is oxidized in the dark, and the (Q-+F-+) back reaction regenerates the photoactive F P680 Q state. The electrochemical and kinetic evidence is consistent with the hypothesis that the Signal IIf species, F, is identical with Z, the physiological donor to P680.


DNA-binding proteins from Novikoff hepatoma cells. In 0.05 M NaCl, 6-8% of the total soluble proteins from Novikoff hepatoma cells bind rapidly and reversibly to columns containing either heterologous or homologous DNA adsorbed to cellulose. These proteins can be eluted by buffer containing 2.0 M NaCl. 0.5-1% of the total protein exhibits a 7-17-fold preference for rat DNA over Escherichia coli DNA. 1-1.5% of the proteins bind DNA so strongly that elution cannot be effected by 4.0 M NaCl but can be accomplished by deoxyribonuclease I treatment of the columns. DNA-binding proteins eluted by 2.0 M NaCl were labeled with 125I or 131I and characterized by sodium dodecylsulfate-polyacrylamide gel electrophoresis and isoelectric focusing. These experiments indicate that DNA-binding proteins represent a discrete subset of the total soluble protein. Many similarities were noted between the major components of the homologous and heterologous DNA-binding fractions.


Adsorption of small molecules to bovine serum albumin studied by the spin-probe method. A spin-probe technique is used for quantitative EPR studies of adsorption of small molecules on globular proteins, of the rigidity of their binding to the protein and of the polarity of the environment. In the case of bovine serum albumin it is shown that nitroxyl radical (2, 2, 6, 6-tetramethyl-4-oxy-1-oxyl-piperidine)-stearate (I) has an adsorption behaviour similar to that of the fatty acids, nitroxyl radical (2, 2,4, 4-tetramethyl-1, 2, 3, 4-tetrahydro-5, 6-benzo-gamma-carboline-3-oxyl) (II) to that of the tryptophan molecule. Radical I rotates relative to the protein molecule, while Radical II is rigidly bound to the protein.


Insoluble non-collagenous cartilage glycoproteins with aggregating sub-units. Two insoluble non-collagenous glycoprotein fractions (A and G) have been separated from puppy rib cartilage, following extraction of most of the proteoglycan and digestion of the insoluble residue with purified collagenase. After reduction, alkylation and extraction with sodium dodecylsulfate most of each protein is solubilized. Gel electrophoresis of solubilized A or G shows the presence of either one or two bands and gel chromatography shows both high and low molecular weight peaks. The production of a low molecular weight electrophoresis band from the high molecular weight Sephadex fraction indicates that there is aggregation and disaggregation of sub-units in sodium dodecylsulfate. Both A and G are high in aspartate plus glutamate and have a low hydroxyproline content. The insoluble A and G both contain hexose, uronic acid, galactosamine, glucosamine and a small amount of sialic acid, but they differ in their contents of hexose and six amino acids. They both form single bands in CsCl gradients but they differ in density. Electron microscopy shows that both insoluble glycoprotein fractions stain with lead, ruthenium red, or alcian blue plus phosphotungstate and that G contains many fine filaments. Material with the same appearance and staining properties was found to occur on the surface of collagen fibres in the undigested cartilage residue.


A simple and inexpensive method for the preparation of (beta,gamma-32P) guanosine 5'-triphosphate. A rapid and inexpensive method has been developed for the synthesis of 32P-labeled guanosine 5'-triphosphate (GTP). When yolk platelets isolated from brine shrimp cysts are incubated with 32PPi at pH 5.8 and in the presence of 10 mM MgCl2 and 5 mM dithiothreitol, the primary compound formed is [beta,gamma-32P]GTP. The synthetic reaction is catalyzed by the yolk platelet enzyme, GTP : GTP guanylyltransferase, which has been demonstrated to be important in the biosynthesis of diguanosine 5'-tetraphosphate (Gp4G), the major purine nucleotide in brine shrimp yolk platelets and encysted embryos.


Three DNA polymerases of rat ascites hepatoma cells: properties of the enzymes and effect of RNA synthesis on the reactions. Three different DNA polymerases have been isolated from rat ascites hepatoma cells [1--3]. The molecular weight of a DNA polymerase (polymerase C) purified from the soluble fraction of the cells was estimated to be 142 000 by sedimentation on a sucrose gradient, while the molecular weights of two DNA polymerases (polymerase P-1 and P-2) purified from nuclear membrane-chromatin fraction were estimated to be 117 000 and 44 000, respectively, by the same method. Under certain conditions, the poly (dT) strand of poly[(dA)-(dT)] was copied well by the polymerases, especially by the nuclear polymerases. Poly (dC) was a good template for the high molecular weight DNA polymerases C and P-1, but poly(dT) and poly(dA) were not effective templates. By addition of complementary oligoribonucleotides, the single-stranded deoxypolymers were copied by the high molecular weight polymerases C and P-1. When single-stranded fd phage DNA was used as template, the polymerization reactions by the high molecular weight polymerases were stimulated by the concomitant synthesis of RNA. This indicates that the oligoribonucleotide acts as a primer in these reactions.


Participation of manganese ions complexed with tRNA in the interaction with amino acids and dipeptides. Carboxyl groups of glycine, beta-alanine, gamma-aminobutyric acid and diglycine interact with Mn2+ coordinated by tRNA, as revealed by 1H and 13C NMR studies. The amino groups of these compounds interact with tRNA phosphate groups. The distances between the coordinated Mn2+ and carboxyl groups and the alpha-protons of glycine were determined. The role of Mn2+ and Mg2+ complexed with nucleic acids in the formation of specific complexes between proteins and nucleic acids is discussed.


Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. A cell-line derived from a patient with chronic myelogenous leukemia (CML) is described. The new cell-line, which has over 175 serial passanges in a 3 1/2-yr period, has the following characteristics: (1) CML cells started to proliferate actively since they were first incubated in culture media. A threefold increase in the total number of cells was observed during the first seven passages; the cell population increased by a factor of 10 to 20 every 7 days from passage 8 through 85; from 20 to 40 times from passage 86 through 150, and more than 40 times after 150 passages. (2) The majority of the nononucleated cells are undifferentiated blasts. (3) The karyotype of all the cells examined show the Philadelphia (Ph1) chromosome and a long acrocentric marker plus aneuploidy. The Giemsa-banding studies identified the Ph1 chromosome as a terminal deletion of the long arm of chromosome 22:del(22)(q12) and the long acrocentric marker as an unbalanced reciprocal translocation of one chromosome 17 and the long arm of one chromosome 15. (4) The CML cells do not produce immunoglobulins, are free of mycoplasma, Epstein-Barr virus, and herpes-like virus particles. (5) CML cells have no alkaline phosphatase and myeloperoxidase activities and did not engulf inert particles. (6) Cultured CML cells provide a constant source of a specific antigen. This CML cell-line represents a unique source of CML cells with meaningful indicators of malignancy for clinical and experimental studies.


Hemoglobin J-Chicago (beta76(E20) Ala yields Asp): a new hemoglobin variant resulting from substitution of an external residue. An electrophoretically fast-moving hemoglobin variant was found in a 2-yr-old boy who was referred for evaluation with findings of iron deficiency anemia. The anemia was corrected, and no hematologic abnormality remained after treatment with iron. Oxygen affinity of the blood was normal, and no evidence was found of instability of the variant hemoglobin. Structural studies demonstrated a substitution of aspartic acid for alanine at beta76 (E20). This change did not appear to cause any functional disruption of the hemoglobin in this patient, as would be predicted by the position of the affected animo acid residue on the surface of the molecule.


Mechanism of the protective effect of succinic semialdehyde and its derivatives in hypoxia. Preliminary injection of succinic semialdehyde into animals in experiments with hypoxia led to a lower concentration of free ammonia in their brain tissues than in control experiments in which the compound was not given. The enzyme system of the liver tissue catalyzes the oxidation of NAD-H-2 by succinic semialdeyde and its dervitives with a free adelhyde group.The conversion of succinic semialdeyde and its derivatives in the liver tissue leading to the formation of the oxidized form of NAD,which is deficient in hypoxia, and the prevention of accumulation of free ammonia in the brain tissue by preliminary administration of succinic semialdehyde are the possible mechanisms of the antihypoxic properties of succinic semialdehyde and its derivatives.


Ultrasound monitoring of hepatic metastases during chemotherapy. Grey-scale ultrasonography has been found to be the most sensitive method of detecting metastatic disease of the liver. In two cases the results of chemotherapy were monitored by ultrasound; the response to treatment could be distinguished from non-response and ultrasonography gave useful information when chemotherapy made radioisotope examination unreliable.


Long-term potentiation of the perforant path-granule cell synapse in the rat hippocampus. When the entorhinal cortex is electrically stimulated, a monosynaptic evoked potential is produced in the ipsilateral hippocampus. Twenty-four hours after a 120-pulse stimulation train, the response to a single test pulse was found to be increased. Stimulation trains presented at a rate of 1/day had a cumulative effect on both the population EPSP and spike components of the evoked potential, and the potentiation so produced lasted at least 12 days. In a few cases, retested 2 months after the last stimulation train, the population EPSP was found to be still highly potentiated while the population spike had declined to initial levels. The most reliable results and largest potentiation were obtained with diphasic stimulation trains of 10 Hz or greater, though some changes were observed even with very low frequencies (0.2 Hz). The results imply that the efficiency of synaptic transmission in the perforant path-granule cell synapse can be modified.


Cellulase location in Cellvibrio fulvus. The location of cellulase in C. fulvus depends on the carbon source for growth and the age of the culture. When cells were grown on glucose or cellobiose all CMC-hydrolyzing enzyme was cell-bound but only part of the activity was located on the cell surface. Treatment of cells with EDTA, lysozyme, and detergents and subsequent fractionation experiments showed that cellulase was also located in the periplasm and bound to a membrane fraction. Growth on cellulose gave cell-free cellulase active against CMC. The enzyme was repressed by glucose but formed at a constant differential rate on cellobiose and amylose. This rate was 8-10 times lower than on cellulose and possible reasons for this are discussed.


Simplified procedures for releasing and concentrating microorganisms from soil for transmission electron microscopy viewing as thin-sectioned and frozen-etched preparations. A simplified procedure is presented for releasing and concentrating indigenous microbial cells from soil for viewing by transmission electron microscopy as thin sections or replicas of frozen-etched preparations. This procedure is compared with two others reported earlier, and their relative merits are discussed as concerns the choice of procedure for the cellular information desired from the soil. Freeze-etching showed that the cell types and size distributions for cells which have been released and concentrated from soil are in general agreement with those for cells in a crude soil slurry in which no attempt to release and concentrate cells was made. Microcolonies were present both in the crude slurry and in the discard soil debris centrifugation pellets from the cell release and concentration procedures. In contrast to the historic assumptions, these microcolonies, as well as some individual cells embedded in soil debris could not be broken up and (or) dislodged so that they would be washed from the soil. The relative numbers of these cells remaining with the soil debris, however, could not be quantitated in the present study.


The ultrastructure of the cellulolytic bacterium Cellvibrio fulvus. Thin sections of the cellulolytic bacterium Cellvibrio fulvus were investigated before and after treatment with polymyxin B and Triton X-100. The Gram-negative vibrio appeared to have no separate stained g2 layer between the L and C membranes. Cells lysed after treatment with polymyxin or swelled in the presence of 5% sucrose. Blebs were formed from the L membrane, and the inner structure of the cells changed. Triton X-100 lysed cells even in the presence of 20% sucrose. The flagellum disappeared before lysis; one of the membranes was dissolved; and no organized inner structure could be seen.


Virusemenia after cytomegalovirus mononucleosis. Cytomegalovirus (CMV) was demonstrated for 15 months in semen from a male homosexual with a recent history of a mononucleosis-like illness. The male urogenital tract can harbor CMV for long periods, and sexual transmission of the virus is more likely in persons with multiple heterosexual or homosexual contacts.


A multiple chemotherapeutic approach to the management of hepatoblastoma. A preliminary report. In 1970, a staging based on surgical resectability of hepatic tumors was devised. Adjuvant chemotherapy with vincristine, 5-fluorouracil, and cyclophosphamide has been given to seven recent cases. Objectives of this study were to set up unified clinical staging and followup; to evaluate the effect of combination chemotherapy on survival in advanced disease; and to evaluate early adjunctive combination chemotherapy in surgically resectable lesions to, hopefully, prevent metastasis. Results to date in seven patients are: no change in the poor prognosis of the three female patients presenting with Stage III-IV hepatocellular carcinoma; the three males with Stage I-II hepatoblastoma have done well and survive the free of disease at 47 months, 44 months, and 28 months; one patient with hepatoblastoma had lung metastasis at diagnosis and died at 7 months with tumor. No toxicity was noted with the use of adjunctive combination chemotherapy following major hepatic resection.


A clinical trial of the oral form of 4'-demethyl-epipodophyllotoxin-beta-D ethylidene glucoside (NSC 141540) VP 16-213. A clinical trial of the oral form of VP 16-213 (NSC-141540), a semisynthetic podophyllotoxin, was undertaken. In 20 patients, treatment was started at 200 mg/day p.o. for 5 days; courses were repeated after a rest period of 16 days. Five patients were treated at the same dose, repeated with only 9-day rest periods. Subsequently, 65 patients were given 300-400 mg/day for 5 days, with rest periods of 9 days between courses. The side effects encountered included anorexia, nausea and vomiting, stomatitis, diarrhea, leukopenia, thrombocytopenia, alopecia, and pruritus. Substernal discomfort with or without palpitations was reported by 18 patients; no explanation for this symptom could be found. No complete remissions (CR) were observed. Parital remissions (PR) and improvement (IMP) were seen as follows: small cell carcinoma, lung (10 patients)--2 PR, 3 IMP; adenocarcinoma, lung (4 patients)--1 PR; alveolar cell carcinoma, lung (1 patient)--1 IMP; mesothelioma (4 patients)--1 IMP; ovarian cancer (12 patients)--3 PR, 3 IMP; breast cancer (20 patients)--4 IMP; colon cancer (8 patients)--2 IMP; bladder cancer (4 patients)--2 IMP; histiocytic lymphoma (7 patients)--2 PR, 3 IMP; chronic myeloid leukemia (1 patient)--1 IMP.


Yoshi 864 (NSC 102627) 1-propanol, 3, 3'-iminodi-dimethanesulfonate (ester) hydrochloride: a phase 1 study. Yoshi 864 was given i.v. push daily times 5 with 6 weeks' followup. Dose escalation was from 0.25 mg/kg to 2.7 mg/kg. Toxicity and effectiveness were first seen at 1.5 mg/kg. Twenty-five courses were given to 16 patients at or above this level. In 16 of 22 courses, exclusive of CML, thrombopenia and/or leukopenia occurred. Mean platelet and WBC nadirs occurred on day 24 and 29 with recovery taking 1-2 weeks and 2-3 weeks respectively. Hb fell in 11 courses. At 2.7 mg/kg, nausea and vomiting lasting 6-12 days occurred in 3 of 7 courses; during 5 coures patients slept 20 hours a day, and 1 was comatose for 2 days. Two patients with squamous cell carcinoma and 1 with an unknown primary responded. Both patients with CML had clinical remissions. It is recommended that a cooperative Phase II Study in a broad spectrum of human solid tumors including lymphomas and chronic myelocytic leukemia be undertaken at a dose level of 2 mg/kg.


Radiation enteritis in children. A retrospective review, clinicopathologic correlation, and dietary management. The clinicopathologic features of radiation enteritis are reviewed in 44 children receiving whole abdominal radiation therapy between 1961-1972 at the Institut Gustave-Roussy. Five of 14 long-term survivors (36%) developed severe delayed radiation injury with small bowel obstruction, occurring within 2 months after completion of irradiation. All had previously had acute radiation reaction during therapy. Histologic appearance in the small bowel at the time of delayed radiation injury revealed severe villus blunting, lymphatic dilatation, and moderately dense inflammatory infiltrate. All patients with delayed radiation injury showed marked clinical improvement coincident with a fractionated low-residue, low-fat diet, free of gluten and free of milk and milk products. The abnormal small bowel roentgenographs and small bowel biopsies reverted to a normal appearance in association with the diet. No exacerbation of radiation enteritis has been seen following dietary therapy.


Wilms' tumor and renal cell carcinoma in retroperitoneal teratoma. A patient with a malignant retroperitoneal teratoma that contained Wilms' tumor and renal cell carcinoma is described. The rarity of renal tissue in teratoma, and the possible mode of development of Wilms' tumor and renal cell carcinoma in such a tumor are discussed.


Syndrome of fetal gigantism, renal hamartomas, and nephroblastomatosis with Wilms' tumor. A new case of the fetal gigantism-renal hamartomas-nephroblastomatosis syndrome is described, in which a Wilms' tumor occurred. It is considered that this observation provides strong evidence for the interrelationship between renal dysplasia and renal neoplasia.


Synovial sarcoma of the neck: a followup study of 24 cases. This article reviews the clinicopathologic features and the biological behavior of 24 cases of synovial sarcoma that took origin from the cervical prevertebral connective tissue space and manifested as a retropharyngeal tumor or as a palpable mass in the anterior or posterior cervical triangle. The age of the 24 patients ranged from 10 to 51 years, with a median of 19 years. Ten patients were women and 14 men. Hoarseness or difficulty in breathing or swallowing were the first symptoms in eight patients. The tumors were solitary and ranged from 2 to 10 cm in greatest dimension. Microscopically, all of the cases showed the characteristic biphasic cellular pattern of a synovial sarcoma, with epithelioid and fibrosarcoma-like areas in varying proportions. Synovioblastic origin of the neoplasm was confirmed by the results of histochemical staining procedures and, in 1 case, by the examination with the electron microscope. Of the 21 cases followup information, 12 had died (10 with pulmonary metastasis) and 9 were alive and free of symptoms. Prompt and complete surgical removal is required to prevent complications from recurrent tumor growth or metastasis.


Specific estrogen binding in rat mammary tumors induced by 7,12-dimethylbenz(a)anthracene. Experiments were undertaken with 7,12-dimethylbenz(q)anthracene-induced mammary tumors of the rat to determine whether ovarian-dependent and ovarian-independent tumors could be distinguished on the basis of differences in the estrogen binding capacity of the tumors in vitro and in vivo. Our results confirm reports showing that ovarian-depentent tumors undergo interaction between ovarian-dependent tumors undergo interaction between 17beta-(3H)estradiol and specific estrogen binding components both in vivo and in vitro, as described for other estrogen target tissues. However, our results also demonstrated that certain 7,12-dimethylbenz(a)anthracene-induced tumors, which continue to grow after ovariectomy of the host, contained significant amounts of 17beta-(3H)estradiol bound to cytoplasmic as well as nuclear components. The sedimentation properties of these components were indistinguishable from those of either ovarian-dependent 7,12-dimethylbenz(a)anthracene-induced tumors or rat uterus. The cytoplasmic binding components of both classes of tumors exhibited similar specificities for estrogens. There did not appear to be an absolute correlation between estrogen-binding capacity of a tumor and its growth response to ovariectomy.


Autoantibodies with antilipoprotein specificity and hypolipoproteinemia in patients with cancer. Sera from 151 patients with a variety of cancers were screened for antibody-like activity against lipoproteins. Eighteen % of the sera exhibited activity against autologous and homologous high-density lipoproteins and 3% exhibited activity autologous and homologous low-density lipoprotiens. Antibody-like binding was proven by its restriction to the Fab fragment of IgG.  The reactive part of the lipoprotein molecule was shown to be the appoprotein. Quantiation of the serum lipoproteins indicated that thehigh-density lipoprotien concentration in the sera of cancer patients was significantly lower (psmaller than 0.01) when antibody was present. These observation suggest that autoimmune mechanisms may be responsible for the decreased high-density lipoprotein serum levels in some patients with cancer.


The glycosaminoglycans in human hepatic cancer. A method is proposed for the analysis of glycosaminoglycans that were isolated from human liver, combining cellulose acetate electrophoresis and enzymatic digestion with mucopolysaccharidases. The major constituent of glycossaminoglycans in the healthy liver is heparin sulfate and/or heparin (about 65%), with approximately equal quantities of dermatan sulfate and hyalauronic acid (about 13.5 and 13%, respectively) and a small amount of chondroitin sulfate. These components, especially chondroitin sulfate and hyaluronic acid, are markedly increased in hepatic carcinomas.


Blastomogenic activity of p-hydroxyphenyllactic acid in mice. Blastomogenic activity of p-hydroxyphenyllactic acid was established by prolonged s.c. administration into C57BL/6 and CC57BR mice. Phenyllactic acid was also tested and was not blastomogenic under the same conditions. The preparations in aqueous solutions were injected into the dorsal region twice weekly in 1.5-mg doses. The total dose per mouse was 42 mg p-hydroxyphenyllactic acid or 50 mg phenyllactic acid. The resorptive character and many forms of blastomogenic activity of p-hydroxphenyllactic acid were shown. It induced neoplasms of varied histogenesis (leukemias, adenomas, hepatomas, and tumors of the vascular tissue),including benign and malignant tumors and precancerous conditions of the urniary bladder.


Comparative adenylate cyclase activities in homogenate and plasma membrane fractions of Morris hepatoma 5123tc (h). Homogenate and plasma membrane fractions of Morris hepatoma 5123tc (h) and rat liver were studied with regard to their relative basal activties of adenylate cyclase and to the comparative responsiveness of this enzyme to glucagon, sodium fluoride, epinephrine, prostaglandin E1, and insulin. The basal adenylate cyclase activities of the hepatoma fractions were found to be similar to those of liver at an adenosine 5'triphosphate concentration of 3.2 mM; if the substrate affinity (Km adenosine 5'-triphosphate) of the tumor enzyme is comparable to that of liver, these findings suggest that the reduced basal cyclic adenosine 3':5'-monophosphate levels found to occur in hepatoma 5123tc (h) probably are not due to a decreased basal rate of formation of this cyclic nucleotide. Glucagon (5.6 muM) significantly stimulated adenylate cyclase in both fractions of hepatoma and livers; however, the responsiveness of the tumor enzyme to this hormone was substantially lower than the responsiveness of liver for both homogenate and plasma membrane preparations; i.e., activities were enhanced 18-fold (relative to the basal activity)for liver homogenate compared with only a 6-fold increase for tumor. With the plasma membrane preparations, glucagon increased the activities 5- and 3.5-fold in liver and hepatoma, respectively. Sodium fluoride (10mM), in contrast to glucagon, increased the adenylate cyclase activity to approximately the same extent (about 10-fold) in the liver and hepatoma preparations. Epinephrine (100 muM) enhanced the liver and hepatoma homogenate activites 3- to 4-fold and the hepatoma plasma membrane activities 2-fold; however, the liver plasma membrane activites were not increased. Prostaglandin E1 (56.6 MUM) significantly increased adenylate cyclase activites of liver and hepatoma homogenates (i.e., 1.5- and 3-fold, respectively) but not of the plasma membrane preparations. Insulin (0.7 muM) did not significantly alter adenylate cyclase activities in any of the preparations.


Increased activity of low-Km cyclic adenosine 3':5'-monophosphate phosphodiesterase in plasma membranes of Morris hepatoma 5123tc (h). The total cyclic adenosine 3':5'-monophosphate (cAMP) phosphodiesterase activities as well as the activities of the low- and high-K-m enzyme forms were investigated in homogenates, 100,000 X g supernatants, and plasma membrane fractions of rat liver and Morris hepatoma 5123tc(h); the responsiveness of hepatoma and liver plasma membrane (low-K-m) phosphodiesterases to imidazole (40 mM) and theophylline (5mM) were also compared at cAMP concentrations of 1 and 7.5 muM. The total cAMP phosphodiesterase activities of tumor homogenates and 100,000 X g supernatant fractions were found to be less than one-half those of liver; kinetic studies of homogenates indicated that this finding was largely due to a substantial reduction (53%) in activity of the hepatoma high-K-m enzyme. In contrast, low-Km cAMP phosphodiesterase activities for tumor homogenate and plasma membrane fractions were significantly (50%) higher than liver; this was particularly evident when cAMP concentrations were between 0.5 and 2 muM. Since these concentrations are in the range of basal physiological levels of cAMP in hepatocytes, the present results suggest that the reduced levels of cAMP, previously observed in hepatoma 5123tc (h), are primarily due TO An increased rate of cAMP metabolism by low-Km cAMP phosphodiesterase in plasma membranes of the tumor. Imidazole increased the activity of the low-K-m cAMP phosphodiesterase of liver plasma membranes by 22 (1 muM cAMP) and 38% (7.5 muM camp); tumor activity was enhanced 35 and 50%, respectively, at 1 and 7.5 muM cAMP. Theophylline inhibited the plasma membrane phosphodiesterase activity of liver 79 and 53% at cAMP concentrations of 1 and 7.5 muM, respectively; hepatoma activity was inhibited 82 (1 muM cAMP) and 62% (7.5 muM cAMP).


Estrogen-prolactin dependency in 7,12-dimethylbenz(a)anthracene-induced tumors. Hormonal influences on dimethylbenz(a)anthracene-induced tumor growth were investigated in detail by endocrine ablation and replacement of hormones. The majority of tumors regressed following ablation and most of them were reactivated by subsequent administrations of estrogen (0.1 to 5 mug) or prolactin (2 mg). Increasing numbers of tumors, however, were not stimulated by prolactin when administration was delayed, and a basal level of estradiol (0.01 mug) in addition to prolactin was required for reactivation of tumors. Nafoxidine hydrochloride, a competitor of estrogen at the receptor sites, arrested growth of a large portion of dimethylbenz(a)anthracene-induced tumors in intact animals but failed to retard growth of prolactin-stimulated tumors. On withdrawal of prolactin-nafoxidine, rapid regression of tumor occurred and readministration of prolactin failed to activate most of the tumors for as long as 28 days. Our results give good supporting evidence that estrogen plays a primary role in tumor growth. The interactions of prolactin and estrogen at tumor sites are necessary for regulatory events related to tumor growth.


Alteration of glycine N-methyltransferase activity in fetal, adult, and tumor tissues. Glycine N-methyltransferase activity has been examined in a number of fetal and adult organs, as well as in several rodent hepatomas, using both enzymatic and immunological techniques. In fetal rabbit liver, the activity first appears at a low level at about 20 days postfertilization and rises to high levels after birth, reaching maximum in the adult liver. In fast-growing hepatomas, the activity could not be detected by either enzymatic or immunological assay. It could be detected in the slower-growing hepatomas, but in considerably diminished levels compared with that of normal adult rat liver. Immunoassays gave no evidence for inactive forms of the enzyme in the tissues that had no enzymatic activity. Transfer RNA methyltransferase assays carried out simultaneously showed an inverse relationship to the glycine N-methyltransferase activity. The levels of transfer RNA methyltransferase activity were high in fetal and tumor tissues and lower in normal adult tissues.


Identification of the virions in the in vitro L1210(V) leukemia cell lines by morphological, virological, and immunological techniques. In vitro L1210 (V) cell lines contained abundant intracytoplasmic A-particles, numerous C-type particles, a small number of B-type particles, and occasional intracisternal A-particles. The intracytoplasmic A-particles were incorporated into both spiked (B-type) and smooth-surfaced (C-type) particles formed at the budding site. Both B-and C-type particles also developed by gradual accululation of neucleooid material. The particles, particularly the C-type, exhibited a wide range of densities. The cells showed strong surface immunofluoresence for both murine mammary tumor virus and Gross murine leukemia virus antigens and variable degrees of cytoplasmic immunoflurescence for the protein antigens (p1 to p6) of Rauscher leukemia virus. The cells, the culture supernatant, and the purified virus each gave positive reactions with murine mammary tumor virus and murine leukemia virus antisera by immunodiffusion. The viral particles failed to infect C57BL, C57BL/6 X DBA/2F1 (hereafter called BD2F1), BALB/c, Af,and RIIIf mice. Howver, the cells were highly tumorigenic in BD1F-1 mice, moderately tumorigenic in BALB/c mice, but not tumorigenic in C57BL, Af, and RIIIf mice.


A comparison of the effects of daunomycin and adriamycin on various DNA polymerases. The effects of the anthracycline antiboties, daunomycin and adriamycin, on the DNA-directed activities of DNA polymerases from murine sarcoma virus, rat liver (high-molecular-weight species), Escherichia coli, and Micrococcus luteus were determined. Under all conditions tested, these compounds had greater inhibitory effect against the viral polymerase than against cellular polymerase. The inhibition of murine sarcoma virus DNA polymerase by daunomycin was competitive with respect to DNA. For viral DNA polymerase it was concluded that the inhibition was predominatly caused by the interaction of duanomycin with the primer-template DNA. Also, an appreciable reversal of the daunomycin-induced inhibition of this polymerase by an increase in Mg-2+ concentration is consistent with the conclusion derived by competition experiments. In contrast, the inhibition of both rat liver and M. luteus DNA polymerases was essentially noncompetitive with DNA. Also, bacterial enzymes wer e less sensitive to inhibition by these drugs than the virion polymerase. The strong and preferential inhibiton of viral DNA polymerase is discussed in relation to a differential sensitivity of normal as compared to tumor cells observed in some cell lines.


Heritability of the phytohemagglutinin responsiveness of lymphocytes and its relationship to leukemogenesis. Upon analyses of 59 inbred strains, F-1 hybrids, and congenic-resistant mouse strains, the strain distribution pattern of the stimulation of perippheral mouse lymphocytes by phytohemagglutinin was established using a micromethod. The family of DBA mice were the lowest responders of phytohemagglutinin, whereas the C57 family responded best. Strain PL/J exhibited the best response. The response of lymphocytes to the lectin is governed by more than two but less than five major genes of unknown linkage. No direct association to the H-2 histocompatibility complex was found, although an indirect influence of this locus could not be excluded. All high-leukemia strains are good responders to phytohemagglutinin. None of the low-responder-group strains exhibit spontaneous leukemia. No correlation of the response of lymphocytes to the expression of the type C RNA genome could be established. Cell suspensions from animals exhibiting clinical signs of leukemia responded only weakly or not all to the lectin.


Benzo(alpha)pyrene effects on mouse epithelial cells in culture. The effect of benzo (a) pyrene on the growth in culture of 5 mouse epithelial cell strains was examined. These epithelial cells are highly sensitive to the cytotoxic action of benzo (a)-pyrene. In addition, the activity of the benzol (a) pyrene-metabolizing system, aryl hydrocarbon hydroxylase, is low but highly iducible by the carcinogen. As the sensitivity of a cell strain to the cytotoxic action of benjo (a) pyrene decreased, the inudcibility of the hydroxylase also decreased,. However, a strong correlation could not be found between cytotoxicity and the level of uninduced or induced hydroxylase when the values from different cell strains were compared. These experiments suggest that thehydroxylase is important in determining the sensitivity of epithelial cells to the cytotoxic action of benzo (a) pyrene, but other factors may also modulate this sensitivity.


Peptidyl proline hydroxylase in adult, developing, and neoplastic rat tissues. A sensitive assay system, optimally supplemented with tritiated protocollagen substrate and cofactors, is described which is suitable for determining the peptidyl proline hydroxylase (PPH) content of a wide spectrum of rat tissues. In most tissues, less than 50 percent of the total activity was soluble; the particulate portion of the activity (concentrated in the mitochondrial and microsomal fractions) was doubled by pretreatment with Triton X-100. Among normal adult tissues, lung had the highest total PPH activity (2.4 times that of liver) and small intestine had the lowest (25 percent that of liver). In brain and lactating mammary gland, the activity was similar to that in skin (60 percent of that in liver). Fetal tissues contained 3 to 8 times more PPH than the corresponding adult tissues, and a much lower portion of the total activity was soluble. In four tissues studied in detail (lung, liver, kidney, and brain), the total PPH declined rapidly during the last few days of gestation; brain attained its low adult value before term, whereas the other three tissues continued to decrease in the course of postnatal development. An injection of cortisol to fetal rats enhanced the decline of PPH in lung, liver, and skull. These experiments suggest that during normal differentiation the decline in collagen synthesis is initiated by fetal glucocorticoid secretion which is maximal on the 19th gestational day. PPH activity appears to be a sensitive indicator of neoplastic growth. In renal, mammary, muscle, and hepatic tumors, the PPH activities were 4 to 10 times higher than in the cognate adult tissue. Even in well-differentiated, slowgrowing tumors, the activity was considerably higher than in any normal, mature, or immature tissue, with the exception of the skull and lung of the 19-day-old fetus.


Comparison of the immunogenicity of hamster cells transformed by adenovirus and Herpes simplex virus. Hamsters vaccinated with adenovirus-transformed cells, modified by acetoacetylation or concanavalin A treatment, or with small numbers of living cells were partly or completely protected against challenge with 3 times 10-6 living cells. Treatment of vaccine cells with iodoacetate, Mitomycin C, neuraminidase plus Mitomycin C did not produce efficient vaccines. Herpes simplex virus-transformed cells treated by any of these procedures did not prevent, and frequently even enhanced, the growth of the homologous living cells; enhancement was often greater in female than in male hamsters. Protective and enhancing vaccines did not induce a different level of cell-mediated immunity, as detected by lymphocytotoxicity tests, which were positive for both homologous transformed cells and nontransformed hamster cells. In contrast, specific complement-dependent cytotoxic antibodies active only on adenovirus-transformed cells were induced by the protective acetoacetylated vaccine prepared from adenovirus-transformed cells; these antibodies were not present after nonprotective vaccinations. The appearance of herpes simplex virus tumors was delayed by treatment with the immunostimulant, Levamisole, or by preimmunization with Newcastle disease virus grown in SV40-transformed cells, but not by Newcastle disease virus grown in herpes simplex virus-transformed cells. Thus, only nonspecific treatments were able to impede herpes simplex virus tumor growth, while protection against adenovirus tumor was accompanied by specific cytotoxic antibodies.


Evaluation of irradiation-plus-urethan-induced murine leukemia virus "release" using a new method for quantitation of oncornaviruses in tissues. The quantity of C-type RNA tumor viruses in homogenate-sonicates of thymus-bone marrow tissues of C57BL/6J and RFM/Un mice 10 days after irradiation (X-rays or gamma-rays)-plus-urethan treatments is no greater than that in thymus-bone marrow homogenates from nontreated  control mice. These results indicate that the leukemogenic activity, shown to be present in such thymus-bone marrow homogenates at this time after irradiation-plus-urethan treatment, is not due to change in quantity of C-type viruses as has been proposed. Virus quantity in tissues was evaluated by a new procedure that includes use of a microchamber with the sides situated on rotor radii so as to produce a uniform virus-containing sediment of tissue homogenate-sonicate that is evaluated by electron microscopic examination of this sections cut perpendicular to the membrane surface. Samples containing as little as 105 to 106 viruses can be relatively easily counted. Semipurified or purified viruses can also be counted after mixing with a tissue homogenate-bovine serum albumin diluent.


A mixed-immunoglobulin rosette technique for detection of antibody to feline oncornavirus-associated cell membrane antigen. A mixed-immunoglobulin rosette technique has been developed for the detection of antibodies to feline oncornavirus-associated cell membrane antigens. Lymphoblastoid cells infected with feline leukemia virus were incubated with test sera and then fixed in paraformaldehyde. They were then exposed to a rabbit anti-cat immunoglobulin G serum. Antigen-antibody reactions were detected by mixing the cells with sheep erythrocytes sensitized with cat anti-sheep red blood cell serum; the presence of cat antibody on the lymphoid cells was registered by the formation of sheep red blood cell rosettes around them. The method was shown to be at least 10 times more sensitive than indirect immunofluorescence. A high degree of correlation was shown between the mixed-immunoglobulin rosette and indirect immunofluorescence tests, using both feline leukemia virus-infected cat and dog cells as targets. The results indicate that the tests are likely to be measuring similar reactions. Using the indirect immunofluorescence test we found that 75% of cats with lymphoid neoplasia had no demonstrable antibodies. Twenty-four of such indirect immunofluorescence-negative sera were tested by the mixed-immunoglobulin rosette technique; antibody was shown in at least seven of these sera.


The regulation of serine dehydratase and glucose-6-phosphatase in hyperplastic nodules of rat liver during diethylnitrosamine and N-2-fluorenylacetamide feeding. Changes in the levels of serine dehydratase and glucose-6-phosphatase induced by dietary stimuli or starvation in hyperplastic nodules of rat liver during diethylnitrosamine or N-2-fluorenylacetamide feeding were studied by immuno- and enzyme histochemical methods. The study was performed during carcinogenesis through a combined method of enzyme histochemistry and radioautography. Serine dehydratase was observed diffusely in the cytoplasm of the original hepatocytes in the periportal zone and was induced markedly during diethynitrosamine feeding but only slightly during N-2-fluorenylacetamide feeding. The enzyme was deficient and not inducible in hyperplastic nodules during their developing phase. Later during the feeding period, however, there was an elevation of the level of serine dehydratase and its inducibility with time in the majority of the nodules. A good correlation was observed between serine dehydratase and glucose-6-phosphatase in their elevated levels and response to enviornmental stimuli. There was a minor group of hyperplastic nodules in which the deficiencies of these enzymes persisted and enzyme induction was not observed. A greater number of hyperplastic nodules with persistent enzyme deficiency was seen during diethylnitrosamine carcinogenesis. These results provide further information about the changing biological nature of hyperplastic nodules with respect to their metabolic adaptability and enzyme levels during hepatocarcinogenesis.


Physicochemical characterization of Novikoff hepatoma mitochondrial DNA. Mitochondrial DNA's (mtDNA) isolated from rat liver and the Novikoff hepatoma grown as both solid tumors and cells in monolayer culture were examined by a variety of physicochemical techniques. Buoyant densities in analytical CsCl equilibrium gradients and thermal denaturation profiles revealed no significant differences in base composition among the mtDNA's isolated from liver, tumor, and hepatoma cells. Sedimentation in neurtral and alkaline CsCl showed no differences in mtDNA size. However, tumor and hepatoma cell mtDNA's were slightly smaller and more heterogeneous in size than liver mtDNA when molecular contour lengths were measured in the electron microscope. Based on chemical determinations, neoplastic mitochondria contained four to five times more DNA per mitochondrion than liver. Also, electron microscopy showed the proportion of mtDNA in complex forms (catenated dimers and oligomers) to be much higher in tumor (18%) and hepatoma cells (15%) than liver (4%).


The presence of histone H1degree in human tissues. Histone Hidegree was found to be present in a number of human tissues. It constituted 50% of the total 5% perchloric acid-soluble histone in human breast, 35%in thyroid, 12 to 18% in adrenal, and 7% in parathyroid tissues. The quantities of histone Hldegree in these human tissues were large compared with the amounts found in rat liver (8%), calf thymus (0%), and HeLa cells (0%). Although the quantity of histone Hldegree was found to vary from one type of tissue to another, it was essentially constant in normal, hyperplastic, and neoplastic human thyroid tissues.


Prolactin and estrogen binding in transplantable hormone-dependent and autonomous rat mammary carcinoma. A hormone-dependent subline of the transplantable rat mammary tumor MTW9 contains binding sites for both prolactin and estrogen. Prolactin binding is saturable (K-d similar to 2 times 10-9 M), hormone specific, and destroyed by proteases. By contrast, an autonomous subline derived from the same parent tumor has lost more than 75% of both prolactin- and estrogen-binding sites, although binding affinities for both hormones are unchanged. This reduction in binding sites for both prolactin and estrogen in the autonomous line may result in an incomplete recognition of the tumor cells as a target for the circulating hormones with a subsequent loss of hormone-dependent growth characteristics.


Messenger RNA species synthesized in vitro by the virion-associated RNA polymerase of vesicular stomatitis virus. The virion-associated RNA-dependent RNA polymerase of vesicular stomatitis virus (VSV) synthesizes in vitro two size classes of RNA products similar to those observed in VSV-infected cells. One RNA product sediments at 31S with an approximate molecular weight of 2.1 X 106. The smaller products consist of at least three classes of RNA sedimenting at 17S, 14.5S, and 12S with molecular weights of 0.7 X 106, 0.52 X 106, and 0.37 X 106, respectively. Hybridization experiments show that both the 31S and 12-18S RNA products are complementary to the genome RNA, and that each class is transcribed from different nucleotide sequences. From the molecular weights of the RNA species and the hybridization experiments, it seems that almost the entire VSV genome RNA is transcribed in vitro.


Anesthesia and the golgi apparatus: enzyme activity changes in a golgi-rich fraction of rat liver after a short ether anesthesia. Activity changes of enzymes in isolated rat liver Golgi preparations at different times (1-48 h) after a short ether anesthesia are reported. Activity of galactosyl-transferase showed a slight gradual increase but thiamine pyrophosphatase decreased sharply, and after 24 h increased to above control level. Arylsulphatase-A remained largely unchanged, and B was significantly decreased. Acid phosphatase activity did remain at the control level, but alkali phosphatase showed a gradual and highly significant increase. Five other enzymes representing probable contaminations from other subcellular organelles, have also been assayed. Correlation is sought between the enzyme activity changes and some other metabolic effects of anesthesia.


Effect of insulin on the proliferation of cultured primate arterial smooth muscle cells. Smooth muscle cells were grown from thoracic aortas of 1-year-old monkeys (Macaca nemistrina). The effect of insulin on the proliferation of these cells was studied by comparing the growth of cells in culture medium to which insulin had been added with that of cells in basal (1% monkey serum) medium and in growth-promoting 5% monkey serum. Insulin in concentrations of 10, 100, 1,000, and 10,000 muunits/ml resulted in successively greater stimulation of growth which was highly significant (P smaller than 0.001) by analysis of variance. There was a significant linear relationship between the logarithm of the insulin dose and cell growth. However, the highest concentration of insulin produced only 50% of the effect of 5% monkey serum. Serum from which insulin had been removed stimulated growth less well (P smaller than 0.05) than did untreated serum at the same concentration (5%) but had significant (P smaller than 0.05) stimulating properties compared with whole serum at a lower concentration. Cells that were older in culture life (eight or nine passages) did not show a growth response to insulin and had an attenuated response to 5% serum. The effect of insulin (100 muunits/ml) was inhibited by dibutyryl cyclic adenosine monophosphate (db-cAMP) (5 times 10-5 M), although there was a latent period of 3 days before inhibition occurred; db-cAMP had no effect on cell counts in the absence of insulin. The electron microscopic appearance of the cells was unaltered by insulin.


In vitro acetylcholine biosynthesis in normal and failing guinea pig hearts. Choline acetyltransferase activity, which is rate limiting in acetylcholine biosynthesis, was measured in the four heart chambers of guinea pigs subjected to (1) sham surgery, (2) constriction of the ascending aorta, (3) constriction of the descending thoracic aorta, and (4) constriction of the pulmonary artery. After 30 days when hypertrophy and heart failure were fully established, choline acetyltransferase was quantified in vitro by a radiochemical assay. In the sham-operated group, enzyme activity expressed in terms of unit weight of cardiac tissue was greatest in the right atrium and the right ventricle and lower in th left atrium and the left ventricle (3.62 plus or minus 0.30, 2.96 plus or minus 0.52, 1.64 plus or minus 0.15, and 1.67 plus or minus 0.22 nmoles/min g-1, respectively). Enzyme activity was reduced (P less than 0.05) in the right atria and the right ventricles of guinea pigs with constriction of the pulmonary artery (1.68 plus or minus 0.37 and 1.31 plus or minus 0.29 nmoles/min g-1, respectively). Enzyme activity also tended to be reduced in the left atria and the left ventricles of guinea pigs with constriction of the aorta. These changes represented a relative dilution of enzyme activity per unit weight but not an absolute depletion, since choline acetyltransferase activity per ventricle was not reduced. The absence of significant changes in the total amount of the neuronal enzyme, choline acetyltransferase, per ventricle contrasted with the observed increases in the myocardial enzyme, carnitine acetyltransferase. These results confirm the presence of significant parasympathetic innervation of the ventricles as well as the atria but do not demonstrate alterations in parasympathetic neurotransmitter biosynthesis in hypertrphied and failing myocardium. The absence of absolute reductions in choline acetyltransferase activity in hypertrophied and failing ventricle contrasts strikingly with the previously reported reductions in tyrosine hydroxylase, which is rate limiting in sympathetic neurotransmitter biosynthesis.


Vitiligo and dysgammaglobulinemia. A case report and family study. This report concerns an 8 year old female with vitiligo and dysgammaglobulinemia characterized by absent IgA, very low IgG, and normal IgM. The t-cell immune system was intact but other family members had low levels or absence of IgA. The possible relationship of dysgammaglobulinemia and vitiligo is discussed along with the classification and inheritance of the immune cell defects.


Control of phosphorylative activity in human liver mitochondria through changes in respiratory enzyme contents. 1. Oxidative phosphorylation and respiratory enzyme activities were measured in the mitochondria from the non-involved lobe of the liver in eighteen patients with massive tumour mainly localized to one lobe and from the regenerating livers of partially hepatectomized rats treated with chloramphenicol. 2. In patients, the concentrations of cytochrome a(t) varied from 40 to 170 pmol/mg of protein. In mitochondria with cytochrome a(t) concentrations more than 70 pmol/mg of protein, the phosphorylative activity per mg of mitochondrial protein was considerably higher than in controls. The mitochondrial oxidative and phosphorylative activities per unit of cytochrome a(t) were negatively correlated with the concentration of cytochrome a(t). These patients with mitochondrial cytochrome a(t) exceeding 70 pmol/mg of protein tolerated partial hepatectomy well. On the other hand, in patients with mitochondrial cytochrome a(t) less than 60 pmol/mg of protein, phosphorylative activity was very low and there was a high surgical mortality. 3. In the regenerating liver of rats treated with chloramphenicol, the oxidative and phosphorylative activities per unit of cytochrome a(t) were negatively correlated with the concentration of cytochrome a(t). 4. It is suggested that an increase in ATP-synthesizing activity per unit of respiratory assemblies is the most basic homeostatic mechanism maintaining energy production in response to an increased metabolic load upon hepatic cells.


The effect of 1,25-dihydroxycholecalciferol on renal tubular reabsorption of phosphate, intestinal absorption of calcium and bone histology in hypophosphataemic renal tubular rickets. 1. 1,25-Dihydroxycholecalciferol (1,25-(OH)2-D3) was given at a dose of 5-0 nmol (2-1 mug) daily by mouth for 4-12 days to three patients with hypophosphataemic (type I), vitamin D-resistant rickets. 2. 1,25-(OH)2-D3 increased intestinal absorption and urinary excretion of calcium without significant effect on the renal handling of phosphate or its plasma concentration. 3. It is concluded that in this type of vitamin D-resistant rickets the renal phosphate abnormality is unlikely to be due to diminished endogenous production of 1,25-(OH)2-D3. 4. The difference between this condition and other hypophosphataemic states is discussed.


Feedback regulation of 25-hydroxycholecalciferol metabolism by vitamin D3. 1. In vitamin D-deficient chicks both vitamin D3 and 1alpha-hydroxycholecalciferol markedly decrease renal 1-hydroxylase activity and induce 24-hydroxylase activity. 2. Actinomycin D abolishes both effects. 3. These results are consistent with feedback regulation of vitamin D3 metabolism by a direct nuclear action of the vitamin or its metabolites on the kidney cells.


Effect of long-acting thyroid stimulator on serum concentration of luteinizing hormone in the rat. 1. The effect of long-acting thyroid stimulator (LATS) on the serum luteinizing hormone (LH) levels of the rat in pro-oestrus has been studied. 2. The injection of three out of four LATS-containing immunoglobulin G fractions caused an increase in amounts of serum LH. 3. Adrenalectomy and dexamethason suppression did not alter this response. 4. Injection of large doses of adrenocorticotrophic hormone did not produce any increase in serum concentrations of LH. 5. It is postulated that LATS may have a direct effect on the release of LH from the pituitary gland.


Disorders of movement: a mercifully short primer. The best known biochemical pathway in the brain is probably the nigrostriatal system. Even so, its relationship to Parkinsonism is incompletely understood. Its relationship to Huntington's chorea and the dyskinetic responses is even less well understood. The significance of this system in other related disorders such as Heterogeneous system degeneration, progressive supranuclear palsy, and striatonigral degeneration is still more speculative. In an effort to elucidate underlying neuropharmacology, trials with levodopa, amantadine, haloperidol, and lithium have been undertaken. This increasing concentration in biochemical mediators reflects changing patterns in neurology where new emphasis on biochemical pathways is replacing in current interest the emphasis previously placed on more traditional neuro-anatomical pathways such as the classical ascending sensory and descending motor tracts.


[Use of the macrophage-electrophoretic-mobility test in the diagnosis of tumours of the central nervous system (author's transl)]. The macrophage-electrophoretic-mobility (MEM) tests of Field and Caspary demonstrates the macrophage-slowing factor (MSF) produced by specifically sentised blood lymphocytes after incubation with the homologous antigen. Besides other diseases with a cellular immune reaction like multiple sclerosis malignant tumours show a reduction of the macrophage mobility in the cell electrophoresis. A significant inhibition in the MEM- test was found in 30 out of  33 tumours of the CNS. Glioblastomas, astrocytomas, spongioblastomas, ependymomas, neurinomas, meningiomas, and a myxofibroma showed a positive result (more than 10% inhibition). The results in ectodermal tumours (pituitary adenoma, craniopharyngioma) differed; secondary brain tumours showed the greatest inhibition.


[The vaso-active intestinal polypeptide in Verner-Morrison syndrome]. A 40-year-old man with the Verner-Morrison syndrome (refractory watery diarrhoea and hypokalaemia in islet-cell tumour of the pancreas) had an islet-cell carcinoma of the non-B-cell type with metastases to liver and mesenteric lymph nodes.  High concentrations of a vasoactive intestinal polypeptide (VIP) were demonstrated in tumour tissue and plasma by radioimmunological tests.  After surgical removal of the tumour the plasma VIP concentration fell to normal.  Immunohistochemical tests of the tumour for gastrointestinal hormones demonstrated marked fluorescence only for vaso-active intestinal polypeptide.  It is assumed that this polypeptide is largely responsible for the clinical signs.  In the described case the severe watery diarrhoea ceased after glucocorticoid administration.


Differences in corticosterone and dexamethasone binding to rat brain and pituitary. In an attempt to relate binding of 3H-corticosterone and 3H-dexamethasone to their respective potencies in blocking pituitary-adrenal activity, cytosol binding in vitro and cell nuclear binding both in vivo and in tissue slices in vitro were studied in hippocampus, hypothalamus, and anterior pituitary of adrenalectomized rats. It was found that the extremely potent glucocorticoid dexamethasone has a different pattern of binding than corticosterone in the brain and in the anterior pituitary. 1) In cytosol, differences in the estimated binding capacities in a particular tissue for 3H-corticosterone and 3H-dexamethasone and different rates of inactivation in the ability to bind the two steroids are observed. 2) For 3H-corticosterone, cytosol binding in hippocampus is higher than that in hypothalamus, and cell nuclear binding follows the same pattern. For 3H-dexamethasone, cytosol binding is again higher in the hippocampus than in hypothalamus but cell nuclear binding in the two structures is not significantly different. With respect to the anterior pituitary, binding to cell nuclei is higher for 3H-dexamethasone, while the binding to cytosol macromolecules is higher for 3H-corticosterone. 3) In vivo and in vitro cell nuclear binding for both steroids showed the same pattern among the three tissues, but in vivo data showed more distinctly the preference of 3H-dexamethasone for the anterior pituitary and the preference of 3H-corticosterone for the hippocampus. 4) When labeled in tissue slices, cell nuclear radioactivity appears to be bound to macromolecules. 5) Steroid metabolism does not occur in slices during 60 min in vitro at 25 C and cannot account for the observed tissue differences in binding. The existence of more than one population of corticosteroid-binding sites in brain and in anterior pituitary is suggested. The results are consistent with the view that the dexamethasone blockade of stress-induced ACTH release is mediated by the anterior pituitary, while the high specificity of cotricosterone binding in the hippocampus implies a specific but as yet undetermined effect of the hormone in this brain area, an effect which may not be directly related to regulation of ACTH secretion.


Rat sertoli cells: a rapid method for obtaining viable cells. A method is described for obtaining populations of viable Sertoli cells from rat testes. Minced whole testes from rats of 15 to 29 days of age are sequentially treated with collagenase and pancreatin. The resulting suspension of cells is sedimented through a sucrose density gradient. Preparations are produced consisting of from 60% to 82% Sertoli cells, an enrichment of 2 to 5 times the proportion of Sertoli cells in whole testes of these ages. The preparations are free of interstitial cells, are essentially free of peritubular cells and contain reduced numbers of germinal cells; the main contaminating cell types are spermatogonia and spermatocytes. The Sertoli cells are considered to be 95% viable by their ability to exclude trypan blue and by subsequent culturing in vitro. The entire procedure requires 3 h. Maintenance of the Sertoli-enriched fraction in modified Eagle's minimal essential medium temporarily at 41 C allows preparations of Sertoli cell monolayer cultures consisting of 95%-98% Sertoli cells within 3 days.


A protein from polymorphonuclear leukocytes (LEM) which affects the rate of hepatic amino acid transport and synthesis of acute-phase globulins. A proteinaceous secretion from phagocytizing polymorphonuclear leukocytes, termed "leukocytic endogenous mediator" (LEM), has been shown to have marked effects on hepatic amino acid transport and RNA and protein synthesis. A single injection of LEM results in a marked accumulation of labeled nonmetabolizable model amino acids in the liver of normal rats. The LEM-stimulated uptake of amino acids by liver was observed in adrenalectomized, hypophysectomized, thyroidectomized, or diabetic rats and could not be duplicated by pharmacological doses of a large variety of hormones. In addition, LEM stimulated an increased uptake of alpha-aminoisobutyric acid by isolated livers during their perfusion in vitro. LEM also stimulated an increased incorporation of orotic acid into hepatic RNA of intact rats, especially into the bound ribosomal fraction. This increased synthesis of RNA preceded an enhanced hepatic production of a number of the acute-phase plasma globulins. LEM did not stimulate the adenylate cyclase-cAMP system in liver and was not found to utilize this system as a second messenger. Thus, the effects of LEM in stimulating hepatic amino acid transport appear to be direct, without mediation by other hormones, and to be independent of cAMP. On the other hand, the ability of LEM to stimulate RNA and acute phase globulin synthesis in liver may require the presence of physiological quantities of hormones such as adrenal corticoids.


Adenine nucleotides in the secretory granule fraction of rat islets. Concomitant with glucose-induced insulin release, there occurred an increase of ATP from 4.40 plus or minus 0.21 to 23.16 plus or minus 0.52 pmol/100 islets/min (P less than 0.001) in the effluent from perifused rat islets. There is a linear relationship between circulating ATP and insulin levels both in the stimulated and basal state (r = 0.689, P less than 0.01). Islets incubated with labeled adenine for a short period of time (37.5 min) showed no release of radioactivity upon subsequent glucose-induced insulin release. Islets incubated for a prolonged interval with labeled adenine (150 min) showed an increase in acid soluble radioactivity in the effluent during glucose-induced insulin release. Following incubation of the islets with labeled adenine for 150 min, approximately 5% of the homogenate radioactivity was found in the secretory granules. Using column chromatography to separate the adenine nucleotides, the distribution of radioactivity among the various nucleotides in the secretory granule fraction was found to be: AMP 54.42 plus or minus 4.96%, ADP 14.20 plus or minus 1.63%, ATP 15.39 plus or minus 3.84%, and cAMP 16.07 plus or minus 2.11%. The distribution of radioactivity in the effluent adenine nucleotides after glucose-induced insulin release was: AMP 32.83 plus or minus 4.62%, ADP 24.52 plus or minus 2.77%, ATP 28.13 plus or minus 5.45%, and cAMP 26.01 plus or minus 3.34%. The absolute levels of adenine nucleotides in the secretory granules were ATP 4.19 plus or minus 0.88, ATP madp 7.94 plus or minus 2.20 and cAMP 4.46 plus or minus 1.74 pmol/ug prot. The levels in the islet effluent were ATP, 15.30 plus or minus 2.70, ATP qDP, 29.43 plus or minus 3.49 and cAMP 7.66 plus or minus 1.93 pmol/100 islets/min for the first ten min of glucose-stimulated insulin release. Thereafter there was a rapid decline in effluent cAMP while ATP and ADP remained in essentially equivalent amounts. The distribution of radioactivity and absolute levels of the adenine nucleotides in the effluent reflects that found in the secretory granules, confirming previous observations that insulin release is occurring by exocytosis.


Genetic studies on the mechanism of action of aldosterone in mice. Aldosterone reduced the urinary sodium:creatinine ratio and increased the potassium:creatinine ratio in a CBA strain of mice. In the Peru strain, under the same conditions, aldosterone had no detectable effect on the sodium:creatinine ratio but had an even more marked effect on the potassium:creatinine ratio. Thus, the effect of aldosterone on potassium excretion is separable from and is not simply a secondary consequence of the effect on sodium retention. Specific binding of 3H-aldosterone to kidney was reduced in Peru mice compared to CBA. However the correlation between 3H-aldosterone binding and physiological effect of the hormone on sodium excretion disappeared totally in a second generation cross between the strains. A possible explanation is that the nuclear binding of 3H-aldosterone as measured here was heterogeneous. In this same second generation cross, the ratio of bound nuclear 3H-aldosterone extracted by low-ionic strength Tris, to the residue of nuclear 3H-aldosterone resistant to extraction by 0.4M KCl, was strongly correlated to the ratio of the effect of aldosterone on potassium excretion to that on sodium excretion. Thus in these experiments the type of nuclear binding was correlated to the type of physiological effect. These results imply that the specific binding of aldosterone to renal tissue was, in some way at least, related to the mechanism of action of the hormone; and that the heterogeneity of this binding had functional significance in terms of the separable effects of aldosterone on renal sodium retention and potassium excretion in these two strains of mice.


Evidence that the pars intermedia and pars nervosa of the pituitary do not secrete functionally significant quantities of ACTH. We have compared the capacity to secrete ACTH in response to stress or adrenalectomy in control rats and in those with total hypophysectomy (H), adenohypophysectomy (AH) with preservation of the intermediate and the neural lobes, neurohypophysectomy (NH) with removal of the pars nervosa and all or part of the pars intermedia with preservation of the adenohypophysis, or incomplete adenohypophysectomy (IAH) in which a portion of the adenohypophysis and all of the pars intermedia and pars nervosa were left intact. Plasma ACTH measured with an N-terminal antibody that reacts on an equimolar basis with ACTH and alpha-MSH but not with other known pituitary hormones was elevated after ether or tourniquet stress in all except the H group. Three weeks after adrenalectomy there was an elevated basal plasma ACTH and an augmented ACTH response to stress in intact and IAH but not in AH rats. When a more specific alpha11-24 ACTH antibody was used there was a high plasma ACTH after ether stress in the IAH, NH, and intact groups but not in the AH or H groups. Adrenal weight and plasma corticosterone after tourniquet or ether stress were indistinguishable in the AH and H groups and were much higher and nearly identical in the intact, NH and IAH groups. We conclude that only the adenohypophysis secretes functionally significant amounts of ACTH. Plasma ACTH detected by the N-terminal antibody in the AH group is probably related to alpha-MSH or similar peptides and is incapable of maintaining adrenal weight or stimulating corticosterone secretion.