In vivo transfer of cholesteryl esters from high density lipoproteins to very low density lipoproteins in man.

The fate of cholesteryl esters in high density lipoprotein (HDL) was studied to determine whether the transfer of esterified cholesterol from HDL to other plasma lipoproteins occurred to a significant extent in man. HDL cholesteryl ester, labelled in vitro with [3H] cholesterol, was injected into human subjects. Labelling of cholesteryl esters in very low density (VLDL) occurred rapidly and by 3 h, the esterified cholesterol in VLDL reached peak specific radioactivity. The removal rate of cholesteryl esters from HDL appeared to be exponential and of the order of 0.2/h; calculation of the apparent flux was about 150 mg/h which approximates reported values for total cholesterol esterification in human plasma in vivo. The rapid rate of labelling of VLDL from HDL suggests that the transfer of HDL cholesteryl esters to VLDL may represent a significant pathway for the disposal of HDL cholesterol.


Different developmental changes in latency for two functions of a single membrane bound enzyme: glucose-6-phosphatase activities as a function of age.

(1). The capacity for the synthesis of glucose 6-phosphate from PPi and glucose as well as for glucose-6-P hydrolysis, catalyzed by rat liver microsomal glucose-6-phosphatase, increases rapidly from low prenatal levels to a maximum between the second and fifth day, then slowly decreases to reach adult levels. When measured in enzyme preparations optimally activated by hydroxyl ions, the maximum neonatal activities were 4--5-fold higher than in adult animals and several-fold higher than had previously been observed for the unactivated enzyme. (2) The latencies of two catalytic activities associated with the same membrane-bound enzyme show strikingly different age-related changes. The latency of PPi-glucose phosphotransferase activity reaches high levels (60--80% latent) soon after birth and remains high throughout life, while the latency of glucose-6-P phosphohydrolase decreases with age. The phosphohydrolase is 2--3 times more latent in the liver of the neonatal animal than in the adult. (3). The well established neonatal overshoot of liver glucose-6-phosphatase is almost entirely due to changes in the enzyme in the rough microsomal membranes. The enzyme activity in the rough membrane reaches a maximum and then decreases after day 2, while that in the smooth membrane is still slowly increasing. Despite the great differences in absolute specific activities and in the pattern of early enzyme development between the rough and smooth microsomes, enzyme latency in the two subfractions remains parallel, glucose-6-P phosphohydrolase being only slightly more latent, while PPi-glucose phospho-transferase is much more latent in smooth than in rough membranes throughout life. (4). Kidney glucose-6-P phosphohydrolase and PPi-glucose phosphotransferase activities were found to change in a parallel fashion with age, showing a small neonatal peak between days 2 and 7 before rising to adult levels. Kidney phosphotransferase activity, like that of liver, remained highly latent throughout life. In contrast to liver, the glucose-6-P phosphohydrolase of kidney did not show a characteristic decrease in latency with age and in the adult remained appreciably more latent than in liver. (5). An improved method was devised for the separation of smooth microsomes from liver homogenates.


Inhibition of K+-sensitive p-nitrophenylphosphatase activity on Rhesus-positive red cells after incubation with IgG-anti-Rhesus-D.

The incubation of ghosts derived from human Rhesus-positive red cells with IgG-anti-Rhesus-D inhibited the K+-sensitive p-nitrophenylphosphatase activity. This enzyme has a partial function in the (Na+ + K+)-ATPase system related to the phosphorylation step, which is important for active potassium transport through the red cell membrane. The specificity of the impairment by the antigen-antibody reaction in the Rhesus-D system was proved by the following controls. Ghosts obtained from Rhesus-negative red cells incubated by IgG-anti-Rhesus-D and those of Rhesus-positive red cells treated with non-immune serum did not show any reduction of the K+-p-nitrophenylphosphatase activity. The ghost preparation with lanthanum carried out after hypotonic hemolysis of the washed red cells in 2 mM LaCl3 at pH 6 was the most suitable procedure to explore this topic in comparison to other techniques for preparing ghosts of red cells.


Protein phosphorylation in rat caudate homogenate: stimulatory effects of dopamine and enhancement of dopamine response following 6-hydroxydopamine treatment.

Dopamine (3-hydroxytyramine) stimulates the incorporation of 32P into proteins endogenous to a homogenate of rat caudate nucleus when 10 microM [gamma-32P]-ATP is used as a substrate following preincubation with 400 microM ATP. The increase in 32P incorporation has pharmacological characteristics similar to caudate tissue. Chronic depletion of striatal dopamine in vivo by stereotaxic injection of 6-hydroxydopamine in the nigrostriatal pathway results in a significant enhancement of the dopamine stimulation of 32 p incorporation in vitro. Cyclic AMP-stimulated phosphorylation of caudate proteins remains unchanged following 6-hydroxydopamine treatment.


Change of isozyme pattern during activation of a transforming gene product: its relation to other biochemical markers of cellular transformation and differentiation.

Isozyme patterns of leucine aminotransferase were studied in connection with glucose transport and DNA synthesis during the activation and deactivation of the transforming gene product in rat kidney cells transformed by one Rous sarcoma virus mutant (which has a temperature-sensitive lesion in its transforming gene. On temperature shift-down of confluent transformed cells grown at 40 degrees C in the presence of fresh serum, isozyme III of leucine aminotransferase appeared in 12--20 h, with increasing amounts from 24 to 48 h. Upon temperature shift-up, isozyme I became the predominant form in these cells within 4 days, the major change occurring within the first 24 h. The rate of protein turnover was similar to the rate of loss of isozymes I and III during temperature shift-down and shift-up, respectively. A stimulation of incorporation of [3H]thymidine into DNA was observed within 8--12 h after temperature shift-down of the transformed cells. For the maintenance of stimulated DNA synthesis for at least 16 h, continued exposure to the permissive temperature is not necessary. Stimulation of glucose transport occurred prior to the stimulation of [3H]thymidine incorporation. The isozymes of leucine aminotransferase also changed during the in vitro differentiation of Yaffee L6A cells in such a way that isozyme I represented the major part of this enzyme in the fused myotube, and isozyme III was more predominant in the less differentiated state (mononucleated cells).


A heat-stable protein inhibitor of cyclic 3',5'-nucleotide phosphodiesterase.

A heat-stable, non-dialyzable inhibitory factor of cyclic nucleotide phosphodieterase was detected in and partially purified from bovine retina. The factor appears to be a protein, since the inhibitory activity was abolished by trypsin digestion but not by DNAase or RNAase treatment. The protein inhibitor from bovine retina effectively inhibits the Ca2+-independent phosphodiesterase from several sources, including bovine retina, bovine rod outer segment, and a human lymphoblastic leukemia cell line, indicating lack of tissue and species specificity.


Amylase release from rat parotid glands. I. General characteristics.

The involvement of calcium, ATP, and cyclic AMP-dependent protein kinase activity in the release of amylase from rat parotid glands was examined. Pretreatment of the glandular tissue in 11.25 mM Ca2+ medium potentiated the secretory responses to: dibutyryl cyclic AMP, elevation of the extracellular K+ concentration, reduction of the H+ concentration, La3+, and caffeine. Uncoupling of oxidative phosphorylation blocked release induced by dibutyryl cyclic AMP, K+, and reduction of H+, but had no effect on La3+, caffeine or tolbutamide-stimulated release. Inhibition of cyclic AMP-dependent protein kinase activity blocked only dibutyryl cyclic AMP-induced release and did not inhibit the responses to K+, reduction of H+ or caffeine. The loss of lactate dehydrogenase was used to access the integrity of the tissue during amylase release. No significant increase in the release of lactate dehydrogenase was observed during the secretory responses to: dibutyryl cyclic AMP, La3+, caffeine, or tolbutamide. Triton X-100 and ethanol increased the efflux of both amylase and lactate dehydrogenase. The differential involvement of Ca2+, ATP, and cyclic AMP-dependent protein kinase activity in amylase release induced by the various secretagogues suggests that three types of reactions are involved in the release of amylase.


A Leydig cell tumour: a model for the study of lutropin action.

The properties of cells isolated from a Leydig cell tumour have been compared with normal rat testis Leydig cells. These cells were found to be similar in the following respects: 1. Lutropin-stimulated cyclic AMP and testosterone production. 2. Lutropin-activated protein kinase activity followed by phosphorylation of endogenous proteins of mol. wts. 57,000and 14,000. 3. Parallel lutropin dose vs. response curves for phosphorylation of the endogenous proteins and for testosterone production. 4. Two forms of isoenzyme, cyclic AMP dependent protein kinase, present. They differed mainly with respect to the lutropin-stimulated testosterone production, which was much lower in the tumour cells compared with the normal adult testis Leydig cells (4.6 +/- 1.1 and 114 +/- 16 ng testosterone/10(6) cells per 2 h, respectively). However, the lutropin-stimulated steroid production in the tumour cells was quantitatively comparable with the normal rat Leydig cell when the metabolism of pregnenolone in intact cells and mitochondria was inhibited by addition of SU-10603 and/or cyanoketone. It is concluded that the Leydig cell tumour used in this study can be used to investigate certain aspects of lutropin action where large quantities of cells are required.


Role of cytoskeletal elements in cytochalasin E-induced superoxide production by human polymorphonuclear leukocytes.

The release of superoxide anions from human polymorphonuclear leukocytes induced by cytochalasin E was greatly enhanced by the pretreatment of the cells either with deuterium oxide or with concanavalin A. Colchicine, vinblastine and cyclic AMP inhibited the release. Cytochalasins A and B also suppressed the superoxide release. These observations suggest the involvement of microfilament-microtubule system in the production and release of superoxide anions induced by cytochalasin E.


Comparison of cyclic nucleotide specificity of guanosine 3',5'-monophosphate-dependent protein kinase and adenosine 3',5'-monophosphate-dependent protein kinase.

Guanosine 3',5'-monophosphate-dependent protein kinase (cyclic GMP-dependent protein kinase) and adenosine 3',5'-monophosphate-dependent protein kinase (cyclic AMP-dependent protein kinase) exhibited a high degree of cyclic nucleotide specificity when hormone-sensitive triacylglycerol lipase, phosphorylase kinase, and cardiac troponin were used as substrates. The concentration of cyclic GMP required to activate half-maximally cyclic dependent protein kinase was 1000- to 100-fold less than that of cyclic AMP with these substrates. The opposite was true with cyclic AMP-dependent protein kinase where 1000- to 100-fold less cyclic AMP than cyclic GMP was required for half-maximal enzyme activation. This contrasts with the lower degree of cyclic nucleotide specificity of cyclic GMP-dependent protein kinase of 25-fold when histone H2b was used as a substrate for phosphorylation. Cyclic IMP resembled cyclic AMP in effectiveness in stimulating cyclic GMP-dependent protein kinase but was intermediate between cyclic AMP and cyclic GMP in stimulating cyclic AMP-dependent protein kinase. The effect of cyclic IMP on cyclic GMP-dependent protein kinase was confirmed in studies of autophosphorylation of cyclic GMP-dependent protein kinase where both cyclic AMP and cyclic IMP enhanced autophosphorylation. The high degree of cyclic nucleotide specificity observed suggests that cyclic AMP and cyclic GMP activate only their specific kinase and that crossover to the opposite kinase is unlikely to occur at reported cellular concentrations of cyclic nucleotides.


Rat pancreas adenylate cyclase. VI. Role of the enzyme in secretin stimulated enzyme secretion.

1. The responsiveness of adenylate cyclase and enzyme secretin for secretin and the C-terminal octapeptide of pancreozymin has been investigated in particulate fractions of the pancreas of five different species. 2. The adenylate cyclase is sensitive to the C-terminal octapeptide of pancreozymin in all species investigated. 3. The enzyme is much more sensitive to secretin in rat and cat than in mouse and rabbit, whereas with guinea pig intermediate values are obtained. 4. The enzyme secretion is stimulated by secretin in pancreatic fragments of rat and cat, but not in those of mouse and rabbit. 5. These results suggest that in species where secretin stimulated enzyme secretion, it does so by stimulating the adenylate cyclase system.


Transferrin receptors during rabbit reticulocyte maturation.

Experiments were performed to examine the fate of transferrin receptors in reticulocytes as these cells mature in vivo to erythrocytes. Reticulocytosis, synchronized by administration of actinomycin D, was induced in adult rabbits. Simultaneous measurements were made of haematological parameters and the interaction between transferrin and reticulocytes while the cells matured in vivo to erythrocytes. As the reticulocytes matured there was a parallel decline in their ability to take up transferrin and transferrin iron. At the same time, there was a proportionate decrease in the density of receptors for transferrin on the reticulocyte surface. The affinity of the receptors for transferrin remained unaltered during the maturation process. It was concluded that the inability of erythrocytes to take up transferrin or its iron is due primarily to the loss of transferrin receptors from the maturing reticulocyte surface.


Stimulation by polyamines of carbamylphosphate:glucose phosphotransferase and glucose-6-phosphate phosphohydrolase activities of multifunctional glucose-6-phosphatase.

The effects of added polyamines on carbamylphosphate (carbamyl-P):glucose phosphotransferase and glucose-6-phosphate (Glc-6-P) phosphohydrolase activities of rat hepatic D-Glc-6-P phosphohydrolase (EC 3.1.3.9) of intact and detergent-treated microsomes have been investigated. With the former preparation, in the presence of 1.4 mM phosphate substrate and 90 mM D-glucose (phosphotransferase), 1 mM spermine, spermidine, and putrescine activated Glc-6-P phosphohydrolase 67%, 57%, and 35%, respectively. Carbamyl-P:glucose phosphotransferase, under comparable conditions, was activated 57%, 34%, and 18%. NH+4 (0.25--5.0 mM) produced at best but a minor activation (0--14%), while poly(L-lysine) (Mr = 3400; degree of polymerization 16) equimolar relative to other polyamines with respect to ionized free amino groups activated the hydrolase 358% and the transferase 222%. Treatment of microsomes with the detergent deoxycholate reduced, but did not abolish, polyamine-induced activation. The stimulatory effects of polyamines persisted in the presence of excess catalase, indicating their independence from H2O2 formation; and were eliminated in the presence of Ca2+. Kinetic analysis revealed that all tested polyamines decreased the apparent Michaelis constant values for carbamyl-P and Glc-6-P, but had no effect on the Km for glucose. Poly(L-lysine) increased the V value for both Glc-6-P phosphohydrolase and apparent V values for phosphotransferase extrapolated to infinite concentrations of either carbamyl-P or glucose. The other tested polyamines elevated only this last velocity parameter. It is proposed that a major mechanism by which polyamines activate glucose-6-phosphatase-phosphotransferase is through their electrostatic interactions with phospholipids of the membrane of the endoplasmic reticulum of which this enzyme is a part. Conformational alterations thus induced may in turn affect catalytic behavior. It is suggested that polyamines, or similar positively charged peptides, might participate in the cellular regulation of synthetic and hydrolytic activities of glucose-6-phosphatase.


Chromosome aberrations in B lymphocytes of atomic bomb survivors.

Although chromosome aberrations in T lymphocytes and bone marrow cells have been reported in atomic bomb survivors, the presence of chromosome abnormalities has not been demonstrated in B lymphocytes because of the technical difficulties involved in B-lymphocyte separation. A method for detecting chromosome aberrations in B lymphocytes was established by "stimulation" of B lymphocytes with Epstein-Barr virus (EBV) instead of "separation" of B lymphocytes by rosette formation. The EBV-stimulated lymphocytes were isolated as single colonies in soft agar and transferred to liquid culture for further cell growth. The EBV-stimulated B lymphocytes of two heavily exposed survivors showed 50% and 12.5% chromosome abnormalities 30 yr after exposure to the effects of the atomic bomb. The former patient seemed to have a karyotypically abnormal clone of B lymphocytes in vivo. The method used in this study and the evidence of chromosome aberrations in B lymphocytes for long periods after radiation exposure will be useful and important in elucidating the malignant processes of acute lymphocytic leukemia, B-cell lymphoma, and multiple myeloma among high-risk groups having a history of accidental or therapeutic exposure to radiation or radiomimetic drugs.


[Free radicals formed during UV-irradiation of unsaturated fatty acids containing peroxidation products].

On the irradiation of oxidized linolic acid an assymetric ESR signal with g-factor 2.0039 and the linewidth 30--33 Oe has been registered. The signal could be the result of photolysis of hydroperoxides. During prolonged irradiation of all the samples accumulation of alkyl radicals characterized by a symmetric signal with a well-defined ultrafine structure has been observed. An increase of the sample temperature up to 87 K in the presence of oxygen was preceded by transformation of alkyl radical R into RO2. All the radicals disappeared at the temperature of 160 K. All these processes seem to be the first phases of photoperoxidation of unsaturated fat acids in lipid-containing systems, including biological membranes.


[ESR study of mitochondria in basal metabolic states and at different levels of ATP-synthetase activity].

Effect of metabolic transitions of mitochondria on the value of ESR-absorption measured at room temperature has been studied. It has been found that the content of free radicals with ESR signal of g = 2.00 and the halfwidth about 15e depends on the metabolic state of mitochondria determined by the load of respiration and phosphorylation systems of mitochondria. The maximal gain of the free radical content is observed in the active state when the maximal load is applied to the phosphorylative chain. An attempt to localize the region of initiation of additional free radicals by means of uncouplers and inhibitors of phosphorylation, as well as by placing mitochondria into the phosphate-free medium has shown that the change in ESR absorption of mitochondrion preparation is in a specific correlation with the change in energization of mitochondria. The analysis of alternative ways for an increase of the free radical fund during energization makes it possible to connect this increase with the work of ATP-syntetase and to suggest that free radicals take part in the ADP phosphorylation by inorganic phosphate, being probably, localized directly on the ADP molecule.


Neutrophil-mediated antibody-dependent killing of herpes-simplex-virus-infected cells.

The ability of neutrophils to serve as effectors of antibody-dependent cell-mediated cytotoxicity (ADCC) was studied using herpes-simplex-virus-infected (HSV-infected) human embryonic lung cells as targets. Killing of infected cells by neutrophils required HSV-immune serum and did not take place in the presence of nonimmune serum or in the absence of serum. Uninfected cells were not killed. ADCC was evident by 30 min and reached a value of 66% by 4 hr at an effector:target ratio of 100:1. Cells from patients with chronic granulomatous disease were as effective as normal cells, and this indicates that ADCC could be mediated by processes that are independent of the respiratory burst.


Interaction between neuronal amine uptake and prejunctional alpha-adrenergic receptor activation in smooth muscle from canine blood vessels and spleen.

Experiments were performed to determine the conditions in which norepinephrine release from adrenergic nerve terminals in smooth muscle from canine blood vessels and spleen might be inhibited by prejunctional alpha-adrenergic receptor activation. Strips of aorta, mesenteric and splenic arteries, splenic capsule and portal and saphenous veins were labeled with 7-3H-norepinephrine and mounted for superfusion. In the portal vein, an inhibitory effect of prejunctional receptor activation with exogenous norepinephrine (1.2 X 10(-6) M) on transmitter efflux could be demonstrated during electrical stimulation (9 V, 2 Hz) of the nerve terminals. By contrast, in the other tissues, inhibition of transmitter release during electrical stimulation or depolarization of the nerve terminals with K+ (40 mEq/l) could only be demonstrated aftet blockade of the neuronal uptake mechanism. That activation of prejunctional alpha-adrenergic receptors in blood vessels inhibits the exocytotic process is suggested by the failure of exogenous norepinephrine to affect either the basal efflux of 3-H-norepinephrine or the displacement of 3H-norepinephrine by tyramine.


Kinetics of alpha-adrenoreceptor blockade by phentolamine in the normal and denervated rabbit aorta and rat vas deferens.

The rates of onset and offset of alpha-adrenoreceptor blockade by phentolamine (5 X 10(-7) M) have been studied on the normal and denervated rabbit aorta and rat vas deferens. Removal of the adventitia with its accompanying sympathetic nerve component results in an approximately threefold increase in the rate of onset of alpha-adrenoreceptor blockade by phentolamine in the rabbit aorta. The rate of offset of blockade by the antagonist on the aorta is likewise faster after the adventitia has been removed (approximately threefold faster for loss of 90% of the antagonist from the region of the receptors). This effect is not likely to be attributed to the absence of sympathetic nerve terminals since surgical denervation of the rat vas deferens has no effect on the kinetics of receptor blockade in this densely innervated organ. These data suggest that the adventitia of the rabbit aorta may serve as a diffusion barrier which limits the antagonist's access to, and efflux from, the region of the receptors. The dissociation constant of phentolamine (calculated from the equilibrium dose ratio) on the alpha-adrenoreceptor is unaltered by denervation in either tissue and is identical in both tissues. It is suggested that alpha-adrenoreceptors in the rabbit aorta and rat vas deferens are of a single type and are not significantly affected, with respect to antagonist affinity, by denervation.


In vitro relaxation of arteries and veins by prazosin: alpha-adrenergic blockade with no direct vasodilation.

Controversy exists regarding the mechanism by which prazosin lowers blood pressure without a marked increase in heart rate; a mechanism involving both sympatholytic activity and direct smooth muscle relaxation has been suggested. alpha-Adrenergic receptor blockade by prazosin is well documented and occurred to exogenous norepinephrine and to field stimulation in vitro in rat arteries and veins. A parallel shift of the norepinephrine concentration response curves in the aorta and mesenteric artery contrasted with a nonparallel shift and a marked depression of maximal norepinephrine responses in the inferior vena cava, portal, iliac and femoral veins. Nonspecific direct acting vasodilators will antagonize contractile responses to all agonists. However, prazosin (10(-8) M) specifically antagonized norepinephrine-induced responses. Concentration response curves to potassium chloride or to serotonin were not affected in these rat tissues. In addition, prazosin (up to 10(-6) M) did not significantly relax aortic tissue previously contracted with potassium chloride or serotonin, whereas the vasodilator, nitroglycerin, produced a clear relaxation. Prazosin only reduced the tone of vessels contracted with norepinephrine. These data indicate that prazosin exhibits minimal, if any, direct smooth muscle relaxant properties in concentrations higher than those producing alpha-adrenergic receptor blockade, and relaxes rat veins by a mechanism involving alpha-adrenergic receptor blockade.


Innervation of the canine inferior vena cava. Distribution of adrenergic and cholinergic excitatory fibers among the embryologically distinct segments.

The innervation of three embryologically distinct segments of the canine inferior vena cava was investigated. These segments were termed A (supradiaphragm), B-C (intrahepatic and that between liver and renal veins), and D (infrarenal). Strips were cut from these segments, and their isometric tensions were recorded. Transmural electrical stimulation induced contractile responses in circular strips from segment B-C (66.5% of the maximum norepinephrine-induced response) and in those from D (14.4%), but not in A. These responses almost completely disappeared in the presence of phenoxybenzamine. In segment B-C, however, the remaining small contraction was markedly enhanced by neostigmine and abolished by atropine. The same phenomenon was also observed in the contraction remaining after reserpinization. Longitudinal strips from segment C responded similarly. Concentration-response curves of circular strips for acetylcholine were shifted by neostigmine markedly to the left only in segment B-C, while no significant shift occurred in A and D. It was concluded that the adrenergic innervation is remarkably dense in B-C, sparse in D, and probably lacking in A. In addition, a cholinergic excitatory innervation is present in segment B-C.


In vivo effect of endrin on three phosphatases in kidney and liver of the fish Ophiocephalus punctatus.

The effect of endrin (0.030 ppm) on the acid phosphatase, alkaline phosphatase and glucose-6-phosphatase activities in the kidney and liver of a teleost fish, Ophiocephalus (Channa) punctatus has been studied. The period of exposure was twenty days. An increase in the acid phosphatase activity is noted in the two tissues examined here. There is no significant change in the activity of alkaline phosphatase in kidney while in liver the enzyme is inhibited. In the kidney glucose-6-phosphatase showed a slight increase in activity but it is inhibited in liver.


Effects of childhood cancer on long-term survivors and their families.

Problems experienced by families of long-term survivors of acute lymphatic leukaemia and Wilms's tumour were investigated to find out the best way of using limited resources to improve management of such patients. All patients had received treatment at Alder Hey Children's Hospital, and all had completed treatment at least two years before the study. A social worker interviewed the parents of each child. The results showed that various aspects of management needed improvement, including: information given to parents at diagnosis of their child's illness and during subsequent treatment; continuity of care and multidisciplinary teamwork among those caring for the child; greater understanding by school teachers that such children have the same educational needs as others; wider communication by hospital staff with the child's other relatives, particularly grandparents; financial help for parents; and marital counselling. To help implement these proposals full-time social workers were attached to the hospital. Preliminary results were encouraging, though it is too early to evaluate the long-term effects of the changes.


Substance P: depletion in the dorsal horn of rat spinal cord after section of the peripheral processes of primary sensory neurons.

The substance P content, glutamic acid decarboxylase and choline acetyltransferase activities and the level of [3H]diprenorphine binding were measured in various regions of the lumbar spinal cord of rats after unilateral section of the sciatic nerve or after dorsal rhizotomy. Sciatic nerve section produced a 75--80% depletion of substance P in the dorsal horn but did not change the substance P content of the ventral horn. The onset of substance P depletion occurred within 7 days and was maintained for 2 months. The substance P content of the dorsal root ganglia and both the peripheral and central branches of primary sensory neurons was also reduced after sciatic nerve section. Glutamic acid decarboxylase and choline acetyltransferase activity were unchanged; however, a small decrease in opiate receptor binding occurred 1 month after nerve section. Dorsal rhizotomy produced an 80% depletion of substance P in the dorsal horn. In addition, the substance P content of the ventral horn was significantly reduced. Glutamic acid decarboxylase activity in the dorsal horn was unaffected by dorsal rhizotomy whereas opiate receptor binding was reduced by 40%. From these studies it appears that peripheral nerve injury results in the degeneration of primary sensory neurons which contain and release substance P as neurotransmitter.


Infundibular localization of vasopressin, oxytocin and neurophysins in the rat; its relationships with corticotrope function.

The distribution of vasopressin (VP), oxytocin (OXY) and neurophysins 1 and 2 (N1, N2) has been studied in the median eminence (ME) of normal, heterozygous (HDI) and Brattleboro (DI) rats. Numerous thin periportal VP and N2 fibres exist in the normal and HD1 rats; they have never been observed in the DI rats. N1 and OXY fibres in the external layer of the median eminence are rare. Adrenalectomy increases periportal VP and N2 loading in normal and HDI rats; it does not modify the appearance of the DI median eminence. Dexamethasone prevents external VP and N2 overloading in adrenalectomized rats injected during the whole postoperative period. Injections of vasopressin indicated that it had a negative feedback effect during a short time (3 days) but not during a longer period (7 days). The suprachiasmatic neurons are stained only by anti-VP and anti-N2 antibodies. Their overstaining induced by adrenalectomy disappears with dexamethasone, aldosterone or vasopressin treatment. This central effect of hormones is not necessarily associated with disappearance of overloading in the external layer of the ME. The hypothalamo-infundibular tract carrying VP and N2 is involved in regulatory mechanisms of the corticotrope axis. Its relationships with suprachiasmatic neurons are discussed.


Cerebellar opiate receptors in lagomorphs. Demonstration, characterization and regional distribution.

The cerebellum of lagomorphs (pika, rabbit, hare) binds 100--200 femtomoles of [3H]etorphine per milligram of protein. This is very high in comparison with the 10--15 fmol/mg found in the cerebellum of rodents (mouse, hamster, rat). In the rabbit cerebellum, the etorphine sites have binding properties indistinguishable from those of genuine opiate receptor sites in brain. They exhibit a high affinity for [3H]etorphine (KD = 1 X 10(-10) M), [3H]naloxone (KD = 9 X 10(-10) M), morphine and levorphanol but not for dextrophan. Moreover, sodium ions enhance binding of naloxone (antagonist response) and diminish binding of etorphine, morphine and levorphanol (agonist response) to cerebellum homogenates. The regional distribution of [3H]etorphine binding sites in the rabbit cerebellum points toward concentrations higher in the neocerebellum (hemispheres) than in the archecerebellum (lingula and flocculonodular lobe). Finally, the specific concentration of opiate receptor sites in the isolated molecular layer is at least two times that in the isolated granular layer and ten times that in white matter.


Effect of nerve activity on the in vivo release of [3H]serotonin continuously formed from L-[3H]tryptophan in the caudate nucleus of the cat.

A new isotopic approach has been developed to study the in vivo release of serotonin (5-HT). 'Encéphale isolé' cats were implanted with a push-pull cannula in the ventrocaudal part of the head of the caudate nucleus to estimate the release of [3H]5-HT continuously synthesized from L-[3H]tryptophan. Both [3H]5-HT and [3H]tryptamine were found in superfusates. Resting steady state in the release of [3H]indoleamines was observed as soon as 20 min after the beginning of the superfusion with L-[3H]tryptophan; the levels of [3H]5-HT in superfusates were 2.5 times those of [3H]tryptamine and about 6 times the blank value. They were markedly enhanced in the presence of fluoxetine (5 x 10(-6)M), a blocker of the 5-HT uptake process. A marked increase in the release of [3H]5-HT was seen during the local depolarization of 5-HT terminals with potassium chloride (60 mM) or batrachotoxin (10(-6)M) or during the stimulation of 5-HT cell bodies in the nucleus raphe dorsalis with L-glutamic acid (5 x 10(-5)M). These treatments did not enhance the efflux of [3H]tryptamine. The potassium-evoked release of [3H]5-HT was reduced by LSD (10(-5)M). LSD added alone in the superfusing fluid was without effect. The batrachotoxin-evoked release of [3H]5-HT was inhibited in the presence of tetrodotoxin (9 x 10(-6)M). The spontaneous release of [3H]5-HT and [3H]tryptamine was markedly reduced in the presence of a calcium-free medium containing cobalt (10 mM). A transient slight reduction in the spontaneous release of [3H]5-HT was observed in the presence of tetrodotoxin (9 x 10(-6)M). The local cooling of 5-HT cell bodies with a cryoelectrode induced a slight reversible decrease in [3H]5-HT release. These last two treatments were without significant effect on [3H]tryptamine efflux in superfusates. These results indicate that the release of [3H]5-HT endogenously formed from [3H]tryptophan is dependent on nerve activity and that this is not the case for [3H]tryptamine. The advantages of the isotopic approach for in vivo studies on the release of 5-HT are discussed.


Chronic relapsing experimental allergic encephalomyelitis: morphological sequence of myelin degradation.

Myelin degradation in chronic relapsing experimental allergic encephalomyelitis was studied using light microscopic histochemistry and electron microscopy. In the earliest stages, a large number of myelin stripping macrophages were found. The Luxol fast blue positive degradation products were then gradually transformed into PAS positive material. No sudanophilic stage of myelin degradation was found in this model. In electron microscopy, the Luxol fast blue positive material was identified as uniformly layered lipid inclusions with a periodicity of 4.0--4.5 nm. During further digestion, this material was transformed into polymorph structured material, consisting of lamellar leaflets with a diameter of 7--10 nm, curved cylindrical profiles and granular osmiophilic material.


Effects of parachlorophenylalanine and 5, 6-dihydroxytryptamine on the free-running rhythms of locomotor activity and plasma corticosterone in the rat exposed to continuous light.

Parachlorophenylalanine (PCPA) and 5,6-dihydroxytryptamine (5,6-DHT), depletors of brain serotonin, were administered to the rat and circadian rhythms of locomotor activity and plasma corticosterone were determined simultaneously in individual rats in light-dark cycles (LD) and in 200 lux continuous light (LL). Free-running periods and acrophases on the 12th day in LL (LL12) were calculated by a least squares spectrum method. In PCPA-treated rats which showed 70% depletion of brain serotonin, circadian rhythms of locomotor activity in LL and of plasma corticosterone and ACTH in LD disappeared for several days after the drug injection. Circadian rhythms of locomotor activity reappeared after the LL7 day and free-ran with a phase shift. Free-running periods of these rats did not differ significantly from that of control rats. However, the acrophase of PCPA-treated group on the LL11 day was 5 h advanced as compared to that of control. Circadian rhythm of plasma corticosterone in the PCPA-treated rats was detected on the LL12 day but their peak times were distributed around 24:00 h instead 08:00 h observed rats. The 5,6-DHT-treated rats which showed only 40% depletion of brain serotonin exhibited normal free-running rhythms in both locomotor activity and plasma corticosterone in LL and no difference in the acrophases of these functions on the LL12 day as compared to controls. These results suggest that PCPA affects the circadian clock (or clocks) itself in such a way that it blocks the clock to free-run or at least it effectively shortens the free-running periods of locomotor activity and plasma corticosterone in the rat.


Spectral studies of human erythrocyte catalase.

The optical absorption and circular dichroic spectra of human erythrocyte catalase (EC 1.11.1.6) and its cyanide, azide, and fluoride derivatives over the wavelength range of 210 to 700 nm are reported. Treatment with acid or alkaline solutions causes spectral changes which may be due to dissociation of the enzyme into subunits and removal of the heme group from the protein. The fractions of the protein structure present as alpha helix, beta pleated sheet, and unordered structure have been estimated from the CD spectrum in the far-ultraviolet region. The CD spectra also indicate that the protein conformation does not change appreciably after cyanide binding. The epr spectroscopy of the native enzyme and its cyanide complex are reported. The spectral results are compared with catalase obtained from other mammalian and bacterial sources.


Gangliocytic paraganglioma of the duodenum.

In spite of possessing microscopic features of invasiveness, the clinical behaviour of duodenal gangliocytic paraganglioma is benign. The operation of choice for removal of the tumour is local excision. This conservative approach was used in the case of a 70-year-old white women who presented with occult gastrointestinal bleeding. The unusual histologic features of this woman's tumour are discussed. Although the tumour appears to be aggressive no recurrence has been reported to date following adequate local excision of a duodenal gangliocytic paraganglioma.


Effects of irradiation on mixed müllerian tumors of the uterus.

A retrospective study of 54 patients with histologically proven malignant mixed müllerian tumors of the uterus was undertaken with main emphasis on the evaluation of the effects of irradiation on pelvic tumor control. The tumors were staged according to the FIGO classification for endometrial carcinoma and 24 were classified as Stage I, 10 as Stage II, 13 as Stage III and seven as Stage IV. Patients with Stage I and II were treated with surgery alone (9 patients, three surviving) or preoperative intracavitary irradiation (13 patients, eight surviving) or preoperative combination of intracavitary and external irradiation (12 patients, six surviving). Five patients with Stage III and IV were treated with surgery alone, two were treated with a combination of irradiation and surgery and 11 with radiation alone. None of these patients survived. In seven patients showing no residual tumor in the uterine specimen after irradiation, no pelvic failures were noted, whereas seven of 17 (41.2%) with residual tumor developed pelvic recurrences. In patients with Stage I treated with surgery alone, three out of six recurred in the pelvis whereas only three of 17 (17%) receiving preoperative irradiation developed pelvic recurrences. However, in Stage II six of eight patients treated with preoperative irradiation failed in the pelvis. Correlation with the doses of irradiation given to the uterus or the pelvic lymph nodes indicate that with doses below 5000 rads a significantly higher number of pelvic recurrences take place, whereas these are uncommon with doses over 6000 rads. The difference in pelvic recurrences between dosage levels is not, however, statistically significant. It is suggested that patients with Stage I and II malignant mixed müllerian tumors of the uterus should be treated with preoperative radiation and total hysterectomy with bilateral salpingo-oophorectomy. Patients with more  advanced disease have extremely poor prognosis and should be treated with radiation therapy alone. This tumor has a high propensity to spread through lymphatics and hematogenous metastases are seen in approximately 75% of the patients. Because of this dissemination, significant improvements in survival rate will not be seen until effective cytotoxic agents are available.


Adjuvant immunotherapy of lung cancer with BCG cell wall skeleton (BCG-CWS).

Four hundred fifty-five patients with lung cancer were treated with oil-attached cell-wall skeleton of bacillus Calmette-Guérin (BCG-CWS) as adjuvant immunotherapy following initial conventional therapy. The overall survival period of the patients was prolonged significantly as compared with that of 380 patients in a historical control group receiving initial conventional therapy alone (p less than 0.0001). The prolongation of the survival period of the patients was also statistically significant when classified according to clinical stages and histological cell types. The therapeutic effect was remarkable in patients combined with malignant pleurisy. Intrapleural injection of BCG-CWS resulted in not only prevention of accumulation of pleural effusion and abrogation of tumor cells but also in prolongation of survival period (P = 0.016). No serious side effects due to BCG-CWS were experienced. From the previous experimental studies and clinical experiences with human tumors, it can be concluded that adjuvant immunotherapy with BCG-CWS is a useful therapeutic modality for lung cancer, especially in cases combined with malignant pleurisy.


Malignant melanotic neuroectodermal tumor of infancy: a clinical, pathologic, ultrastructural and tissue culture study.

The melanotic neuroectodermal tumor of infancy is an uncommon neoplasm typically of early childhood which has a predilection for the head and neck region, particularly the maxilla. Except for one previous example in the literature, this tumor has consistently behaved in a benign fashion. This study documents the clinical course and pathologic findings of a tumor which began in the maxilla of a 4-month-old boy, followed by a local recurrence, metastasis to a cervical lymph node and finally, widespread dissemination and death at 18 months, 24 months and 38 months, respectively. The tumor was initially composed of nests consisting of melanin-containing cells and small dark cells. An elevated vanillylmandelic acid level was recorded during the course of the disease. At autopsy, the tumor in lymph nodes, liver, bone and soft tissues had a monotonous pattern of small dark cells similar to a conventional neuroblastoma. Previous ultrastructural studies indicate that the melanotic neuroectodermal tumor of infancy is composed of melanocytes and neuroblast-like cells. Our case provided the unique opportunity to examine in sequence the ultrastructural and in vitro characteristics of a recurring and eventually metastasizing melanotic neuroectodermal tumor. Although the neuroblast-like cells were initially difficult to identify by electron microscopy, a melanin-producing cell line and a separate nonpigmented cell line were successfully isolated from various tumor explants. Various stages of melanosome development were identified in the pigmented cells from the local recurrences and in vitro. Dibutyryl cAMP accentuated the formation of pigment and dendritic development in the melanocytes and dendrites only in the small nonpigmented cells. Electron dense granules were observed in the cultured smaller cells and also in the lymph node and soft tissue metastases. Tyrosine hydroxylase activity was demonstrated in the neuroblast-like cells. In the final biopsy and autopsy material, only the neuroblast-like cells remained and the tumor resembled a conventional neuroblastoma.


Adenosarcoma of uterus and ovary: a clinicopathologic study of two cases.

We report the clinical and pathologic features of two cases of adenosarcoma. Our first case occurred in the ovary of a 51-year-old woman with extensive pelvic involvement. She was treated vigorously with radiation and chemotherapy and at the present time almost 9 years later the patient is free of disease. This represents a case with one of the longest survival reported in the literature. Our second case occurred in a 37-year-old woman and was limited to the endometrium.


Adenoid cystic carcinoma: biologic behavior in 38 patients.

This study presents the clinical and pathologic data on 38 patients with adenoid cystic carcinoma. Most of the tumors arose from the major salivary glands or the mucus glands of the upper respiratory tract and oral cavity. The typical biological behavior of these tumors was slow but aggressive local growth despite all therapy, followed by eventual death from local disease and/or metastases. Twenty-eight of the patients have died, 22 from adenoic cystic carcinoma, 3 from postoperative complications, and 3 from intercurrent disease. Of the 10 living patients, 4 have active disease, 2 have no evidence of disease 14 years after diagnosis, and 4 have no evidence of disease but follow-up is less than 5 years. We compared various microscopic features of the tumors, including histologic grading, with prognosis but did not demonstrate a correlation in this small series.


Hodgkin's disease terminating in oat cell carcinoma of the lung.

A 27-year-old woman with Hodgkin's disease, nodular sclerasing type, having received two courses of radiation therapy with one year and nine months interval and a prolonged course of chemotherapy with combinations of COPP, MOPP and CVPP, developed oat cell carcinoma of the lung, four years after the initial diagnosis.


Partial remission of advanced adenoid cystic carcinoma obtained with adriamycin: a case report with a review of the literature.

Adenoid cystic carcinoma is relatively uncommon and often originates from the salivary glands. Although distant metastases develop rather frequently no satisfactory form of therapy has been reported. We achieved a partial remission with adriamycin in a patient with advanced metastases of adenoid cystic carcinoma. This case is described, and a short review of the literature, including papers on the treatment of advanced disease, is also given.


Trophoblastic pseudotumor of the uterus: case report and ultrastructure.

A trophoblastic pseudotumor occurring in a young woman 6 months after a normal pregnancy is reported. Ultrastructural investigation demonstrated a close structural relationship between the infiltrating cells and those of the trophoblastic components of the normal human placenta, especially those seen in the primary villi of the developing placenta.


Sweat gland differentiation in mammary adenoid cystic carcinoma.

A breast tumor is described which presented as an exophytic mass, and which by both light and electron microscopic examination had a biphasic histologic composition. In the superficial area adjacent to the epidermis, it showed tubular differentiation similar to a cutaneous tubular apocrine adenoma and salivary basal cell adenoma, and, in the deeper portion, it had the characteristic features of adenoid cystic carcinoma. Their possible interrelationships are discussed, and mammary adenoid cystic carcinoma is briefly reviewed.


Inflammatory fibrous histiocytoma in the left upper abdomen with a leukemoid blood picture.

A case of inflammatory fibrous histiocytoma arising in the left hypochondrium, which presented with a leukemoid reaction and was initially diagnosed as chronic neutrophilic leukemia, is described. The difficulty of interpreting the hematological findings, especially as the spleen appeared to be enlarged, is discussed. The tendency of this highly malignant group of tumors to cause a systemic granulocytosis is commented upon and attention is drawn to the lack of satisfactory treatment.


Food safety and public health: interaction of science and law in the federal regulatory process.

The programs of the Food and Drug Administration (FDA), which operates under a broad mandate of regulatory authority provided by the Congress in the form of the Food, Drug, and Cosmetic Act, demonstrate the way in which science and law interact to protect public health through the regulatory process. In particular, sections 402, 406, and 409 of the Act provide the means for regulating both new and old food products approved for use by the petition process as well as foods which present a potential hazard because of environmental accidents which result in residues of undesirable or dangerous chemical substances. The episodes of foods contaminated with polychlorinated biphenyls (PCBs) or polybrominated biphenyls (PBBs), and the manner in which action levels or guidelines were developed to regulate the allowable levels of these chemicals in foods, describe the pragmatic way in which FDA protects public health by restricting the allowable levels of chemical substances in foods.


Increased cyclic adenosine 3':5'-monophosphate phosphodiesterase activity in the epidermis of phorbol ester-treated mouse skin and in papillomas.

The potent tumor promoter, 12-O-tetradecanoylphorbol-13-acetate, produced a 2- to 3-fold increase in the activity of both the low- and high-affinity forms of cyclic adenosine 3':5'-monophosphate phosphodiesterase activity 13 hr after application to mouse skin. The magnitude of the enzyme induction correlated with the tumor-promoting activity of several doses of 12-O-tetradecanoylphorbol-13-acetate and of other phorbol esters. The induction of the low-affinity phosphodiesterase could be blocked by prior i.p. injection of the microtubule poisons, colchicine and vinblastine. The low-affinity cyclic adenosine 3':5'-monophosphate phosphodiesterase activity of the epidermal component of mouse skin papillomas produced by two-stage tumorigenesis was 3 times that of the surrounding uninvolved epidermis.


Reversal by glucocorticoid hormones of the loss of a fibronectin and probollagen matrix around transformed human cells.

Confluent cultured human skin fibroblasts had an extracellular fibrillar matrix of fibronectin and procollagen. Human skin fibroblasts transformed by SV40 did not have such a matrix. Treatment of transformed fibroblasts with 10(-5) to 10(-8) M dexamethasone and 10(-5) to 10(-7) M cortisol, but not testosterone or progesterone, caused partial restoration of the matrix. Glucocorticoid-treated transformed human fibroblasts can serve as a model for partial reversion toward normal or differentiation of transformed human fibroblasts.


Affinity cytotoxicity with an alcohol dehydrogenase-antibody conjugate and allyl alcohol.

A potent new enzyme-antibody conjugate system for amplifying cytotoxicity was tested in a well-defined model of hapten [2,4,6-trinitrophenyl (TNP)]-substituted tumor cells (HEp2) and purified anti-hapten antibody. Brief treatment of TNP-HEp2 cells with low concentrations (0.05 to 0.74 micrograms/ml) of antihapten antibody-alcohol dehydrogenase conjugate (Ab-ADH) followed by culture in complement-free medium containing nicotinamide adenine dinucleotide and allyl alcohol or 2-fluoroethanol resulted in 15 to 90% cell killing as measured by 5-[125l]iodo'-2-deoxyuridine uptake assay. The importance of the complete enzyme system was indicated by reduced or absent cytotoxicity if Ab-ADH, nicotinamide adenine dinucleotide, or allyl alcohol (or 2-fluorethanol) were omitted. Immunological specificity of the Ab-ADH was demonstrated by reduced or absent cytotoxicity when: (a) HEp2 cells were not coated with TNP; (b) Ab-ADH binding onto TNP-cells was blocked by free hapten (2,4-dinitrophenyllysine); or (c) unconjugated alcohol dehydrogenase and anti-TNP purified IgG anti-2,4,6-trinitrophenyl antibody with NAD+ and allyl alcohol or anti-TNP antibody with complement were used.


Localization of retinoic acid-binding protein in nuclei and the nuclear uptake of retinoic acid.

Retinoic acid-binding protein (RABP) has been detected in the nuclei of chick embryo skin and Lewis lung tumor. The nuclear binding component showed the same ligand specificity and sedimentation value as the cytosol RABP. Whereas pronase completely digested the nuclear binding component, DNase showed 40%, and RNase showed negligible digestive action. Retinoic acid binding to the nuclear RABP was completely inhibited by a mercurial, and the inhibition was reversed by dithiothreitol. The nonspecific uptake of retinoic acid by Lewis drug nuclei and chick embryo skin nuclei was inhibited up to 50% by cytosol RABP. The maximal inhibitory effect produced by cytosol RABP was after 45-min incubation. Incubation of Lewis lung tumor with [3H]retinoic acid resulted in the appearance of nuclear RABP: [3H]retinoic acid in the nuclei. The complex formed was weak, and most of the bound retinoic acid could be removed by dialysis.


Cyclic adenosine 3':5'-monophosphate and protein kinase activity in insulin-dependent and -independent mammary tumors.

Approximately 70% of primary 7,12-dimethylbenz(alpha)-anthracene-induced mammary tumors regressed when (tumor-bearing) rats were made diabetic after treatment with streptozotocin. In the intact animal, cyclic adenosine 3':5"-monophosphate (cAMP) levels of tumors that regressed following the induction of diabetes were initially 4-fold lower than in unresponsive tumors but increased 4-fold during regression. The insulin-independent tumors showed no statistically significant changes. cAMP binding in cytosol of regressing tumors was about 80% above the initial values at 36 hr after therapy but decreased to about 45% 1 week later. On the contrary, the binding capacity of the nuclei showed a 56% increase at 36 hr and increased gradually to about 3-fold 1 week later. Within 36 hr after treatment, total histone kinase activity increased 127% in the cytosol and 153% in the nuclei of regressing tumors. The increment of histone kinase activity was almost totally in the cAMP-dependent component of the enzyme. These changes were not apparent in insulin-independent tumors. The results are interpreted to indicate that mammary tumor regression due to diabetes involves the cAMP system and occurs through a sequence of events similar to those observed during regression induced by either ovariectomy or dibutyryl cAMP (cyclic adenosine 3':5'-monophosphate) treatment.


Effect of growth conditions on the content of the major groups of carbohydrates in chick embryo fibroblasts.

The levels of glycogen, hyaluronic acid, chondroitin sulfates, N-acetylneuraminic acid, all of the monosaccharide components of the glycoprotein and glycolipid fractions, and the monosaccharide pools were measured in cultured chick embryo fibroblasts. Under all growth conditions, the glycogen plus the glucose phosphate pool contained approximately 50% of total monosaccharide content of the cells. However, marked qualitative and quantitative alterations were found in the glycoprotein, glycolipid, and mucopolysaccharide fractions when growing cells reached confluence, when the growth temperature was shifted from 36 to 41 degrees, or when the cells were transformed with Rous sarcoma virus. From 65 to 95% of the total monosaccharide residues in these complex carbohydrates were found in the glycoprotein fraction, while the glycolipids contained only 5 to 10% of the residues, and the mucopolysaccharides contained 5 to 25%. Changes in the complex carbohydrates in normal cells following changes in cell density or growth temperature were so great that they obscured any transformation-dependent changes that might have occurred consistently in the virus-infected cells under different growth conditions.


Morphological and biochemical properties of a new human breast cancer cell line.

DU4475 is a new human breast cell line derived from a cutaneous metastatic nodule from a patient with advanced breast cancer. It has been in continuous culture for more than 1 year and has survived 140 subcultivations. The cells grow in suspension, displaying clusters and free-floating aggregates with serpentine-like outgrowths. The cell line is hypotetraploid with modal chromosome numbers between 86 and 93 and possesses five to six distinctive marker chromosomes. The cells grow readily in athymic nude mice and produce tumors from which cells can be reintroduced to culture in vitro.


Delivery of essential services to alcoholics through the "continuum of care".

To provide for the delivery of essential and comprehensive services for alcoholics, a well-developed continuum of care encompasses the following services: an information and referral, or diagnostic, center; detoxification or receiving center; primary rehabilitation; extended rehabilitation; residential intermediate care; outpatient care; a family program; and an after-care program. All of these services are geared toward the ultimate goal of assisting the alcoholic to live independently in the community and to utilize a variety of positive coping devices while remaining abstinent from beverage alcohol.


Type C retrovirus activation and possible functions in the normal and tumor-bearing host.

The pathological consequences of tumor virus infection, transformation, and tumor development in certain experimental animals is a well-established and accepted fact. More recently, it has been suggested that these viruses may also have a physiological function participating in such processes as cellular differentiation, immune recognition, and embryogenesis. This paper delineates the current information giving some credence to physiological function for type C viruses.


Alcohol-related diseases and carcinogenesis.

Possible mechanisms whereby alcohol abuse and alcohol-related diseases may promote the development of cancer are analyzed. The mechanisms discussed include: (a) contact-related local effects on the upper gastrointestinal tract; (b) the presence of low levels of carcinogens in alcoholic beverages; (c) induction of microsomal enzymes involved in carcinogen metabolism; (d) various types of cellular injury produced by ethanol and its metabolites and their relationship to cancer, particularly in the liver; (e) the nutritional disturbances frequently associated with alcohol abuse. The relationship between alcohol-induced cirrhosis and hepatocellular carcinoma is also discussed, and case histories of patients seen at the Bronx Veterans Administration Medical Center with hepatocellular carcinoma in the absence of cirrhosis are reviewed. Data are presented demonstrating the induction, by chronic ethanol consumption, of microsomal enzymes which convert procarcinogens to carcinogens. These data were derived from experiments in which the ability of microsomes isolated from liver, intestine, and lung tissues of ethanol-fed and control rats to activate several test carcinogens was examined in the Ames Salmonella-mutagenicity test. The hypothesis is presented that ethanol-mediated induction of enzyme systems which activate procarcinogens to carcinogens in various tissues contributes to the enhanced incidence of cancer in the alcoholic.


Phase I and preliminary phase II study of neocarzinostatin.

Neocarzinostatin is a protein antitumor antibiotic isolated from cultures of Streptomyces carzinostaticus var.F41. The drug has undergone extensive clinical trial in Japan, and has been reported active against a variety of human tumors. A phase I and preliminary phase II evaluation of the drug has been performed, using an iv bolus daily x 5 schedule. Ninety-six patients have been treated at doses from 500 to 2250 units/m2/day. Courses were repeated at 4-week intervals if allowed by bone marrow recovery. Dose-limiting toxicity was myelosupppression, which occurred late (median nadir, Day 27). Myelosuppression was more pronounced in patients who had received previous chemotherapy. In nine patients (9%) thrombocytopenia was prolonged (greater than or equal to 45 days) or irreversible. Acute administration of the drug was associated with rigors in approximately half the patients. Gastrointestinal side effects were mild. Three patients had a severe acute reaction resembling anaphylaxis. The maximally tolerated dose for this dose schedule is approximately 2250 units/m2/day. Antitumor activity has been seen in hepatoma and hematologic malignancies. Activity in lung and colorectal carcinoma appears limited with this dose schedule.


BCNU-5-fluorouracil combination therapy for recurrent malignant brain tumors.

Twenty-nine patients with recurrent malignant brain tumors were evaluated. BCNU (180 mg/m2) was administered iv on Day 1 and 5-fluorouracil (1 g/m2/day) was given by continuous infusion on Days 15--17. Courses were repeated every 6 weeks until tumor progression occurred. Before each course, a neurologic examination and radionuclide and computerized tomographic scans were performed. Response and progression were defined as improvement or deterioration, respectively, in at least two of the three tests. Nine of 29 patients (31%) responded to therapy, five of 29 (17%) showed progression, and 15 of 29 (52%) had disease stability. The estimated median time to progression was 27 weeks for all patients, 34 weeks for the response group, and 25 weeks for the stable group. Of all 29 patients, 24 (83%) had tumor progression arrested; 40% with response and 6% with stable disease were continuing therapy after 1 year. The stable disease group was larger and the duration of stability was longer than that seen in previous studies. Evaluated by time to progression, a BCNU-5-fluorouracil combination is superior to either BCNU alone or a BCNU-procarbazine combination.


Cyclic alternating combination chemotherapy for small cell bronchogenic carcinoma.

Sixty-one protocol-eligible patients with small cell bronchogenic carcinoma received cyclic alternating combination chemotherapy with two or three non-cross-resistant drug combinations. No chest or prophylactic brain radiation therapy was used. Twenty-eight months after starting treatment, disease-free survival was 23% for patients achieving a complete response (CR) and 13% overall. Initial treatment consisted of high-dose cyclophosphamide, methotrexate, and CCNU (CMC) for 6 weeks. Patients then received vincristine, adriamycin, and procarbazine (VAP) for 6 weeks. The addition of VAP increased the CR rate from 42% to 74% in limited-disease patients and from 24% to 36% in extensive-disease patients. Half of the patients were randomized to a third combination of VO-16-213 and ifosfamide. These patients were cycled at 6-week intervals through the three drug regimens while the remaining patients were cycled between CMC and VAP. The addition of VP-16-213 and ifosfamide did not increase the CR rate or prolong survival. Only complete responders survived beyond 24 months. Sequential use of non-cross-resistant drug combinations represents one method for increasing the CR rate.


Intensive induction therapy for small cell carcinoma of the lung.

Fifteen patients with extensive small cell carcinoma of the lung and no prior therapy were treated with a chemotherapeutic regimen similar in intensity to the approach used in acute myelocytic leukemia. The patients received intensive induction therapy with cyclophosphamide, adriamycin, and VP-16-213 followed by treatment with a combination of BCNU, vincristine, methotrexate, and procarbazine. The objective response rate was 87% (13 of 15 patients) with three complete responses and ten partial responses. With the exception of one patient, the maximal response to therapy was achieved during therapy with the intensive cyclophosphamide, adriamycin, and VP-16-213 regimen. The three complete responders remain in remission for 159, 351, and 285 days but seven of the ten partial responders have relapsed and five of these have died. There was no unexpected morbidity associated with the intensive chemotherapy despite marked bone marrow suppression. This study demonstrates that very intensive combination chemotherapy can be safely used to achieve a high objective response rate in patients with extensive small cell carcinoma, but the complete response rate is low. An analysis of treatment failures and future directions is presented.


In vitro growth of cutaneous T-cell lymphomas.

It is relatively easy to propagate EB virus-transformed normal and malignant B lymphocytes in vitro. Normal T lymphocytes divide for short periods of time after exposure to many mitogens. Exposure of normal T lymphocytes to lymphocyte-conditioned medium (LCM) permits them to divide for longer periods of time, but permanent cell lines cannot be established. Intial attempts to grow malignant T cells from patients with cutaneous T-cell lymphomas (CTCL) in unsupplemented growth medium resulted in the establishment of four EB virus-transformed B-lymphoblastoid lines. The responses of CTCL cells to mitogens and LCM varied from unresponsive to near normal. Subsequent attempts to grow CTCL cells in medium supplemented with mitogens or LCM resulted in the establishment of two cell lines lacking B-cell markers or EB virus. The cells of both lines initially had the ability to form E rosettes, although one of the lines has lost this ability with passage. The two lines can be distinguished from normal T cells by having some or all of the following properties: long-term proliferation, tumorigenecity in nude mice, gradual loss of dependence on mitogen or LCM, and convoluted nuclear morphology.


Effect of sodium loading and ACTH on blood pressure of sheep with reduced renal mass.

Sodium chloride loading produced a rise in blood pressure in intact sheep which was potentiated by reduction in renal mass. ACTH induced hypertension was also potentiated by reduced renal mass, suggesting a volume component for the hypertension when renal excretory capacity for salt and water is reduced.


Comparative studies of the physical state of the lipid phase of normal and hypercholesterolemic very low density lipoprotein.

Very low density lipoproteins (VLDL) were prepared from the serum of rabbits at various stages of hypercholesterolemia (95--1665 mg cholesterol/100 ml of serum). The most notable chemical change in hypercholesterolemic (hc) VLDL was the greatly increased content of cholesteryl esters and the greatly decreased content of triglycerides, compared to normal (n) VLDL. Structurally, the lipid region of n VLDL possessed a much lower microviscosity than did hc VLDL, when analyzed by fluorescence polarization and pyrene eximer methods. The microviscosity of the redispersed n VLDL lipid extract was considerably greater than the observed in n VLDL; but less than that of hc VLDL. Incorporation of pyrene into the lipid region of n VLDL and hc VLDL allowed assessment of various properties of the surface and hydrocarbon regions of these lipoproteins. Only slight differences were found in the pyrene monomer 3 : 1 fluorescence emission peak ratios, and in the rate constant for quenching of pyrene by O2. However, the quenching rate constant of pyrene by I- and iodoheptane were different for each lipoprotein.


Heterotransplantation of human gastric cancer in nude mice.

Human gastric cancer was transplanted into the peritoneum of nude mice, and the progress of invasion and growth of cancer were investigated. Serial transplantations succeeded in 4 strains of human gastric cancer and one strain of canine gastric cancer induced by N-thyl-N-nitro-N-nitrosoguanidine (ENNG). The five strains grew subcutaneously in nude mice, and both single strain and mixed strain were transplanted into the mouse peritoneal cavity by a surgical procedure. In the single strain, cancer cells demonstrated mucosal and/or submucosal invasion in the gastrointestinal tract. In the mucosal layer, cancer permeation into the lymphatic duct was verified. In the histological examination, each strain of the mixed ones grew back to back with no interferance, showing front formation. The human strain and canine strain co-existed in the mouse. A human strain of poorly differentiated adenocarcinoma showed hematogenous metastasis to the liver. This is the first report that the invasion as one of biological characteristics of the primary human gastric cancer was clearly demonstrated, and also, another important biological characteristics i.e., the hematogenous liver metastasis was manifested in the mixed strain.


Guanosine 3', 5'-cyclic monophosphate level in plasma of patients with cancer and various diseases.

Guanosine 3'5'-cyclic monophosphate (cGMP) in the plasma of normal persons and patients with lung or breast cancer and other kinds of neoplasma or other diseases was determined using radioimmunoassay. In comparison with normal persons, significant elevation occurred in the cGMP in the plasma of patients with various kinds of cancer or renal insufficiency. The average cGMP values in the plasma of eight normal persons, 16 patients with lung cancer, 16 patients with breast cancer, five patients with oesophagus cancer, three patients with liver cancer, three patients with stomach cancer, ten patients with renal insufficiency and two patients with myocardial infarction, were respectively 3.46, 9.05, 5.39, 5.42, 7.33, 11.66, 19.55, and 8.0 pmol per ml of plasma. There was no elevation in the cGMP in the plasma of the patients with other diseases studied.


[Vitamin D3 sulfoconjugate distribution and evolution in pregnant or lactating rats].

24 hrs after intravenous injection of vitamin D3 35S sulfoconjugate the radioactivity in kidney is about twice as much in pregnant Rats as in lactating Rats. It decreases from 24 to 48 hrs in pregnant but increases in lactating Rats kidneys. Hydrolysis of vitamin 3H D3 sulfate is detected in kidney and liver extracts, the ratio of free vitamin D3 on vitamin D3 sulfate is higher in pregnant than in lactating animals. Mammary glands of lactating Rats contain mainly the unchanged vitamin D3 sulfate which can be hydrolysed by new born suckling pups.


Dissociation between the electrophysiological properties and total tissue cyclic guanosine monophosphate content of guinea pig atria.

The purpose of this study was to investigate the role of cyclic guanosine monophosphate (cyclic GMP) in mediating the direct electrophysiological effects of acetylcholine in guinea pig atria. Acetylcholine significantly diminished spontaneous rate of right atria without increasing cyclic GMP content. Reductions in rate following acetylcholine were augmented by pretreatment with physostigmine, but cyclic GMP levels remained unchanged. In left atria, acetylcholine significantly shortened action potential duration within 5 seconds (both with and without physostigmine pretreatment), but cyclic GMP content was not significantly elevated. Cyclic GMP levels in right atria were significantly increased in response to acetylcholine when the Ca2+ content of the buffer was elevated from 1.25 mM TO 2.5 MM; however, reductions in automaticity in the right atria were not augmented in the high Ca2+ buffer. Marked increases in cyclic GMP content were produced by Na nitroprusside superfusion without changing automaticity of right atria or action potential duration of left atria. Finally, both right and left atria were superfused with cyclic GMP analogs (8-bromo cyclic GMP and dibutyryl cyclic GMP) at high concentrations (10(-4)) for 15 minutes without producing significant effects on spontaneous rate or action potential duration. These results failed to show a correlation between total tissue cyclic GMP content and the electrophysiological effects of acetylcholine on guinea pig atria. The reasons for this are either that cyclic GMP does not mediate directly the electrophysiological effects of acetylcholine, or that small changes in cyclic GMP concentrations, undetectable when total tissue nucleotide levels are measured, occur in discrete effector pools of the cardiac cell to mediate the intracellular effects of the choline ester.


The combined effect of smoking and coffee drinking on LDL and HDL cholesterol.

Conflicting reports on the effect of smoking and coffee drinking on lipoproteins prompted us to study the combined effect of these two associated, widely prevalent habits in 361 persons randomly sampled from the Evans County cohort. Low-density lipoprotein (LDL) cholesterol levels were significantly higher among persons who smoked cigarettes and consumed five or more cups of coffee per day than among nonsmokers who abstained from coffee. Conversely, high-density lipoprotein (HDL) cholesterol was higher in persons who did not smoke or drink coffee than in coffee-consuming smokers. However, this trend was not statistically significant. Triglycerides and very low-density lipoprotein (VLDL) cholesterol were highest among smokers who drank five or more cups of coffee per day, but these differences did not reach statistical significance. Lipoprotein cholesterol levels were adjusted for age, sex and body mass. Smoking and coffee drinking interact in affecting LDL and total cholesterol, but coffee drinking alone did not appear to affect blood lipids.


Protection of mitochondrial function during ischemia by potassium cardioplegia: correlation with ischemic contracture.

The effect of potassium cardioplegia on mitochondrial function was evaluated in the ischemic isolated rat heart. Mitochondrial function as well as adenosine triphosphate (ATP) levels were determined at the initiation of ischemic contracture, at the completion of ischemic contracture, and 20 minutes following contracture completion. Group I received no cardioplegia prior to ischemia, while Group II received potassium cardioplegia prior to the onset of ischemia. The respiratory control index (RCI), which is the primary measure of the intactness of mitochondrial function, was calculated with both a NAD (nicotinamide adenine dinucleotide)-linked substrate and a FAD (flavin adenine dinucleotide)-linked substrate. Potassium cardioplegia significantly delayed ischemic contracture initiation and completion. Although the RCI and ATP levels decreased significantly at successive levels of contracture, there was no difference in the RCI or ATP content between Group I and Group II at contracture initiation or completion. Unlike previous investigations that have used a time-base to examine mitochondrial function and acute cardiac ischemic injury, we correlated mitochondrial function with the measurable physiologic event ischemic contracture. The data indicated that potassium cardioplegia preserved ATP content and mitochondrial function, and that contracture initiation and completion correlate well with specific ATP levels and mitochondrial respiratory control. The relationship between mitochondrial function and ATP content indicates that the beneficial effect of potassium cardioplegia on mitochondrial function may be secondary to the preservation of high-energy phosphate levels which provide energy for mitochondrial maintenance.


A comparison of angiotensinase and placental leucine aminopeptidase during normal pregnancy.

1. Serum leucine aminopeptidase activity measured by its action on hydrolysis of L-leucyl-beta-naphthylamide was increased progressively as pregnancy advanced. Angiotensinase activity determined by bioasaay showed parallel increase during normal pregnancy. 2. The effects of heat treatment and L-methionine on the activity of angiotensinase were compared with those on leucine aminopeptidase activity in pregnancy serum.


Clinical biochemistry of epilepsy. I. Nature of the disease and a review of the chemical findings in epilepsy.

In idiopathic or generalized epilepsy, serum glucose and cholesterol concentrations tend to be low, especially just before the seizure. Glucose tolerance curves are abnormal and variable. The electrolyte balance is disturbed, and epileptics tend to go readily into alkalosis. Serum [Na+] is usually unaffected, but [K+] is normal to low between attacks and increases during and after the seizure. Serum [Cl-] is usually high just before the seizure. Epileptics are generally mildly hypocalcemic, especially in the period before the seizure. Serum urea and nonprotein nitrogen values are low between paroxysms but increase after the seizure. Serum protein concentration is usually normal. Stress, which releases epinephrine and corticotropin, results in high serum citrate concentration, which probably contributes to decreased serum [Ca2+] just before a seizure. In the healthy individual, any increase in serum citrate is accompanied by increasing [Ca2+]. In the rabbit, convulsions can be induced with corticotropin, a result of increased serum citrate concentration coupled with a decrease in [Ca2+]. The net result is severe hypo-ionic-calcemia. A similar phenomenon has been reported in a few humans. Administration of insulin causes serum citrate concentrations to decrease. Apparently, the dynamic system that controls glucose and lipid metabolism, and thus electrolyte balance, through the hormones epinephrine, corticotropin, insulin, glucagon, calcitonin, and parathormone, is abnormal in the epileptic.


Clinical biochemistry of epilepsy. II. Observations on two types of epileptiform convulsions induced in rabbits with corticotropin.

We propose than an alarm mechanism is operative in animals, designed to regulate neuromuscular irritability by regulating [Ca2+]. Epinephrine or corticotropin (ACTH), injected intramuscularly into animals, causes a hypercitricemia, resulting in decreased [Ca2+]. This increases muscular excitability to facilitate escape. To avoid over reaction, [Cl-] is shifted into the plasma without a concomitant shift of Na+, thus generating an acidosis and an increase in ionization of Ca. Plasma pH, pCO2, total CO2, and [K+] decrease, and [Mg2+] increases. The acidosis, decrease in K+, and increase in [Mg2+] serve to counteract the effect of the decrease in [Ca2+], to protect against tetany. In the rabbit the hypercitricemia observed upon ACTH administration is accompained by a severe hypocalcemia and drop in blood pressure, resluting in tetanic convulsions. This seems to indicate calcitonin release, independent of the hypercitricemia. Thyroidectomized rabbits show only mild hypocalcemia when given ACTH, but develop a severe acidosis and typical grand mal epileptiform seizures. Administration of ACTH and then calcitonin to the goat, an animal resistant to the effects of ACTH alone, simulates the effect observed in the rabbit with respect to changes in blood components and blood pressure. Changes in the blood in the goat and rabbit resemble those in humans before an epileptic seizure. alpha-Melanotropin, containing a portion of the ACTH sequence, reacts in a manner similar to ACTH but more rapidly.


Factors affecting the concentration of triacylglycerols (triglycerides in plasma: reference values for adults.

We studied factors influencing concentrations of triglycerides in the blood of a presumably healthy population of about 9000 persons. Using multi-dimensional analysis, we show, in addition to various related to sex and age, the relation between triglycerides and uric acid, overweight, arterial blood pressure, use of oral contraceptives (in women), consumption of alcohol and tobacco, and lack of physical exercise. These factors were classified as a function of their discriminative power by the statistical method of segmentation. We thus chose the following factors as criteria for exclusion from a reference population for the values of triglycerides in blood: in men 20 to 39 years old, overweight more than 90%; in men 40 to 59 years old, overweight of more than 115%, consumption of more than half a liter of wine per day or of more than 11 cigarettes, use of medicines, and a blood uric acid exceeding 420 mumol/L; in women 20 to 39 years old, contraceptives and overweight; in women 40 to 59 years old, overweight or more than 130% and the intake of medicines. At the 97.5th percentile of the reference population as compared with a non-selected population, the values were 80% less for men 20 to 39 years old, 95% less for men 40 to 59 years old, and only 26 to 30% less for women 20 to 59 years old.


Determination of high-density lipoproteins: screening methods compared.

Factors reflecting the concentration of high-density lipoproteins in serum were assessed for 108 men and 106 women participating in a Venetian screening program for hyperlipoproteinemia. The methods applied, optimized in our laboratory, were: (a) cholesterol in high-density lipoproteins, determined in the supernate after sedimentation of the very-low-density lipoproteins + low-density lipoproteins with dextran sulfate or sodium phosphotungstate; and (b) immunochemical quantitation of apolipoprotein A-I and apolipoprotein A-II by Laurell's "rocket" technique. The latter determinations were performed with total serum before and after delipidation with diispropyl ether/n-butanol (6/4 by vol). The dextran sulfate method gave about 5% higher values than did the phosphotungstate method, but the correlation between the two was excellent (r = 0.95). Results of the immunochemical quantitation indicate that delipidation of lipoproteins before Laurell electrophoresis may not be necessary if only freshly drawn sera are used.


The use of the electroencephalogram in patients admitted for alcohol abuse with seizures.

This study of 174 seizure presenting alcohol abusers revealed that the EEG is overwhelmingly normal by visual inspection. When the EEG is abnormal, however, usefulness is significant. The probable EEG findings in this group are presented and discussed here both clinically and experimentally. The literature is reviewed and while much divergent opinion is revealed, it basically is in agreement with the conclusion as set forth under E, significant findings.


Regression of multiple osseous metastatic renal cell carcinoma.

A 65-year-old man with 6 osseous metastatic lesions was successfully treated by nephrectomy. Five of 6 metastatic tumors, 2 of which were histologically proved, dramatically regressed and disappeared roentgenographically. The other metastasis was removed by amputation 4 years prior to the nephrectomy. No apparent complication was found after the nephrectomy and the patient is in very good health. The factors responsible for regression of osseous metastatic renal cell carcinoma cannot be fully explained. The only common treatment given to 4 patients whose osseous renal cell cancer regressed, was nephrectomy. Since every therapy for metastatic renal cell cancer is unreliable, palliative nephrectomy seems to be justified as a treatment of choice even when osseous metastases are noted and there is a possibility of regression of metastatic lesions. Regression of osseous metastasis from renal cell carcinoma is extremely rare and only 3 cases have been reported.


Adenosine 3',5'-cyclic monophosphate levels in normal and IAA-induced tumours in bean (Phaseolus vulgaris L.) embryos.

Endogenous levels of adenosine 3',5'-cyclic monophosphate in indole-3-acetic acid induced tumours, and control bean embryos were measured with Gilman protein binding assay. No significant differences in cAMP concentrations were found in IAA-induced tumours, which suggests that IAA does not mediate cAMP synthesis in bean embryos.


Chromosome breakage and xenotropic C-type virus in embryonic cultures from New Zealand black mice.

A considerable increase in chromatid and chromosome breaks, as well as excessive fragmentation and "pulverization" of whole metaphase plates was observed in embryonic fibroblast cultures from New Zealand black mice. A C-type RNA virus with a xenotropic host range was isolated from the supernatant fluid of co-cultures of NZB cells and heterologous permissive cells (SIRC cell line). One of the NZB cultures produced this virus without amplification by co-cultivation after spontaneous transformation of the cells. NZB cells are supposed to lack normal restriction of complete xenotropic virus expression and to release this endogenous virus spontaneously at a high level. It is hypothesized that the excessive chromosome damage observed in these cell cultures is related to the permanent production of virus, thus indicating a chromosome breaking effect of endogenous viruses.


Hypersomnolent and nonhypersomnolent patients with upper airway obstruction during sleep.

When the syndrome consisting of sleep-induced apnea and hypersomnolence is due to upper airway obstruction, the hypersomnolence is believed to be the direct result of deprivation of sleep related to such obstructions. The purpose of this report is to describe a group of four asymptomatic subjects with upper airway obstruction during sleep. These subjects were matched with a group of patients with the syndrome of sleep-induced apnea and hypersomnolence. There were no significant differences between symptomatic and asymptomatic groups in terms of the absolute number of upper airway obstructions (252 vs 231), their mean duration (20.8 vs 25.9 seconds), the mean arterial carbon dioxide tension during sleep (39 vs 39 mm Hg), or the electroencephalographic patterns during sleep. The only variables that emerged as significantly different between the two groups were the weights (128 vs 90 kg; P less than 0.05), the low arterial oxygen pressure (PaO2) on waking (54 vs 80 mm Hg; P less than 0.002), and the lower PaO2 during sleep (47 vs 70 mm Hg; P less than 0.01) in the symptomatic patients. From these data, we conclude that the hypersomnolence in patients with sleep-induced apnea due to upper airway obstruction cannot be explained by deprivation of sleep, and other factors need to be carefully examined in future studies.


Biopsy of the main carina; staging lung cancer with the fiberoptic bronchoscope.

The efficacy of main carinal biopsy through the fiberoptic bronchoscope for evaluating resectability has not been determined. Forty-eight patients with carcinoma, but without gross neoplastic involvement of the main carina, underwent biopsy. Five (10 percent) had abnormal results on biopsy of the main carina. Three of the five patients were initially considered candidates for surgery. There were no complications from the procedure. Biopsy of the main carina through the fiberoptic bronchoscope is a valuable staging procedure in selected patients because of its simplicity and yield and the significance of abnormal findings.


Disseminated aspergillosis, Cushing's syndrome, and oat cell carcinoma of the lung.

Disseminated aspergillosis is a known complication of neoplastic disease; however, it is usually associated with neoplasms of the hematopoietic or lymphoreticular systems or with immunosuppressed states. We describe the previously unreported association of disseminated aspergillosis and Cushing's syndrome secondary to a corticotropin-secreting oat cell carcinoma of the lung, and we review the current literature on disseminated aspergillosis complicating neoplastic disease.


Antiviral action of benzo[de]isoquinoline-1,3-diones: 5-nitro-2-(2-dimethylaminoethyl) and 5-nitro-2-[2-(1-pyrrolidine)-ethyl] derivatives.

Two benzo[de]isoquinoline-diones, namely 5-nitro-2-(2-dimethylaminoethyl)-benzo[de]isoquinoline-1,3-dione and 5-nitro-2-[2-(1-pyrrolidine)-ethyl]-benzo[de]isoquinoline-1,3-dione, caused inhibition of the viral replication, when assayed against herpes simplex and vaccinia viruses in chick embryo cell cultures. Influenza and Sindbis virus replication were unaffected by these chemicals. Virucidal effects were unobserved. The inhibitory activity is time-related. Ocular and dermal infections with vaccinia virus in rabbits were prevented or disease severity reduced whenever they were treated with either one of these two drugs.


[A pneumatic colostomy closure].

An atraumatic, pneumatic occlusive colostomy appliance has been described which in animal experiments was well tolerated and was effective in preventing leakage. It is hoped that this may improve the unsatisfactory aspects of management in patients using conventional colostomy appliances.


Malignant histiocytoma of the rectum: report of a case.

A case of malignant histiocytoma of the rectum has been described. We believe this is the first report of a case of this type. Other relevant reports are reviewed.


[Haemofiltration in terminal renal failure (author's transl)].

The concentrations of low-molecular retention products rise during long-term haemodialysis, without any apparent side effects. The authors, having performed haemofiltration in 18 patients who had been on a chronic dialysis programme, conclude that haemofiltration is generally better tolerated than conventional haemodialysis. Haemofiltration benefitted the majority of dialysis-resistant hypertensives, and it also favourably influenced uraemic polyneuropathy. Indications for haemofiltration are thought to be dialysis-resistant hypertension, severe polyneuropathy, and dialysis intolerance.


Effects of adenosine 3',5'-monophosphate, dibutyryl adenosine 3',5'-monophosphate, aminophylline, and imidazole on renal cellular calcium metabolism.

The effects of cAMP, dibutyryl cAMP (DBcAMP), aminophylline, and imidazole on total cell calcium, calcium transport, and distribution were studied in cultured kidney cells by kinetic analysis of 45Ca uptake and desaturation curves. Low concentrations of the cyclic nucleotides (10(-7) and 10(-5) M) increase the total cell calcium, all intracellular exchangeable pools, and calcium transport between all cellular compartments. Aminophylline (1 mM) has effects qualitatively similar to cAMP and DBcAMP, while imidazole has opposite effects. At concentrations of 15 and 40 mM, imidazole depresses the total cell calcium and the cellular exchangeable calcium. Compared to the effects of parathyroid hormone (PTH), the changes obtained with 10(-7) and 10(-5) M cAMP are relatively modest, but higher concentrations (10(-3) M) of both cAMP and DBcAMP produce stimulations as marked as with 15 ng/ml PTH. The most dramatic changes are seen in the mitochondrial calcium pool and in the mitochondrial calcium exchange, which increase between 20- and 40-fold. These experiments show that cAMP mimics the effect of PTH on kidney cells and support the theory that cAMP is the mediator of PTH action on renal cell calcium transport.


Adenosine 3',5'-monophosphate effects on cartilage ribonucleic acid synthesis.

We have previously shown that N6-monobutyryl-cAMP stimulates the incorporation of radiolabeled precursors into the macromolecules of embryonic chicken cartilage incubated in organ culture. The present study assessed the specific effects of N6-monobutyryl-cAMP on RNA synthesis in the cartilage organ culture system. The data show that treatment of cartilage with N6-monobutyryl-cAMP results in a coordinated stimulation of mRNA, rTNA, and tRNA synthesis. Incubation of cartilage with N6-monobutyryl-cAMP causes an increase of [5-3H]uridine incorporation into poly A-containing cartilage RNA and 28S, 18S, and 4S RNA as well. Increased incorporation of the radiolabeled nucleotide occurs without a measurable change in uridine triphosphate specific activity or in the rate of RNA degradation. Further, the isolated poly A-containing RNA directs protein synthesis in an in vitro cell-free system, and stimulation of cartilage mRNA by N6-monobutyryl-cAMP does not alter the size distribution of the mRNA species synthesized from that present in control tissue. These data indicate that cAMP enhances cartilage transcriptional activity. In addition, the increased synthesis of mRNA occurs antecedent to demonstrable effects on protein synthesis. Thus, the effects of cAMP on protein synthesis may be secondary to action at the nucleus.


Daily rhythms in adrenal responsiveness to adrenocorticotropin are determined primarily by the time of feeding in the rat.

These experiments were done to determine: 1) whether feeding-related shifts in daily corticosterone rhythms are dependent upon changes in ACTH rhythms, 2) whether restricted feeding of rats results in abnormally high ACTH and corticosterone levels (i.e. stress), and 3) whether changes in either insulin or glucose levels might be the concomitants of feeding that change adrenal responsiveness to ACTH. Young male rats (80--90 g) on a 12-h light, 12-h dark cycle were allowed access to one of three diets for 2 h/day beginning either at lights off or lights on. The diets contained 3%, 4.5%, or 11% fat. A group of rats had ad libitum access to the food containing 4.5% fat. On day 20 of this regimen, rats were killed at 2- to 4-h intervals during the next 24 h, and plasma ACTH, corticosterone, insulin, and glucose were measured. Adrenal weight and corticosterone content were also determined. In none of these experiments was an ACTH rhythm demonstrable by analysis of variance. Neither ACTH levels nor adrenal and plasma corticosterone levels were higher in animals fed 2 h/day than in rats eating ad libitum. Peak corticosterone levels occurred just before feeding, and the restricted feeding paradigm appeard to sharpen the daily rhythms. However, there was also an effect of the light-dark cycle on corticosterone measures. Dietary fat content was directly related to increases in body weight and mean insulin levels and infersely related to adrenal responsiveness to ACTH. The data show that: 1) the time of feeding determines the timing of the corticosterone rhythm in the absence of a rhythm in ACTH, 2) restricted feeding is not a stress, and 3) neither insulin nor glucose has a substantial influence on adrenal responsiveness to ACTH.


Subcellular localization of thyroxine and reverse triiodothyronine outer ring monodeiodinating activities.

In order to examine the subcellular localization of outer ring T4- and rT3-monodeiodinating activities, nuclear, mitochondrial, microsomal, cytosol, and plasma membrane fractions of rat liver homogenate were incubated with either T4 or rT3 in phosphate buffer (pH 7.35) in the presence of 2 mM dithiothreitol for 15 min at 37 C, and the amount of product (T3 in the case of T4 and 3,3'-diiodothyronine in the case of rT3) was measured by specific RIA. The various tissue fractions were also examined for the relative concentration of various marker enzymes. T4 and rT3 monodeiodinating activities correlated better with enzyme markers of plasma membranes than of any other subcellular fraction in most tissue fractions. A fraction could be isolated, however, in which the monodeiodinating activities correlated better with the enzyme markers of microsomes than of plasma membranes. The various data suggest that plasma membranes and microsomes are two main sites of T4- and rT3-monodeiodinating activities. The location of T4 to T3 converting activity in the plasma membranes may serve to modulate the delivery of the more potent thyroid hormone, i.e. T3, into the cells.


Effects of hypophysectomy, thyroidectomy, and thyroxine on specific prolactin receptor sites in kidneys and adrenals of male rats.

The effect of T4 on specific PRL binding in the kidneys and adrenals was measured in intact, hypophysectomized, and thyroidectomized male rats. PRL binding in the kidneys was reduced from 12.1 +/- 1.1% in the intact group to 4.5 +/- 0.5% after hypophysectomy; 4-day replacement with T4 returned PRL binding to intact values. T4 administered to intact rats significantly increased PRL binding above intact values. Thyroidectomy reduced PRL binding in the kidneys from 14.9 +/- 1.2% to 7.0 +/- 0.6%, and T4 treatment restored PRL binding to intact values. PRL binding was measured at 2, 3, 5, 7, and 10 days after thyroidectomy in kidneys and found to decrease progressively from 8.2+/-0.5% in the intact rats to 2.3+/-0.3% on day 10. A single injection of T4 doubled PRL recetor binding in kidneys of thyroidectomized rats at 12 h and returned binding to intact levels at 24 h. In contrast to the effects of T4 on kidney PRL binding, adrenal PRL binding was only slightly altered by thyroidectomy and T4. These results suggest that T4 has a specific role in regulating PRL receptors in the kidneys but not in the adrenals. Since PRL receptors in the adrenals were only slightly altered by thyroid status, it is believed that the changes in kidney receptors represent a specific rather than a general metabolic effect of T4.


Dissociation of glucocorticoid inhibition of glucose transport and metabolism by increase in adipocyte age and/or size.

Exposure of adipocytes from young rats (2--3 months old) to dexamethasone in vitro results in 40--50% inhibition of glucose transport and metabolism. Comparable experiments with older rats reveal that the hormonal effect on glucose metabolism is progressively reduced to about 20% in 12- to 13-month-old animals and about 5% in 24- to 26-month-old animals. However, no effect of dexamethasone on the glucose transport system can be detected in either of the older age groups when three different sugars are used to measure transport activity. The ability of dexamethasone to inhibit glucose phosphorylation seems to be closely related to inhibition of overall glucose oxidation in both young and old adipocytes. That the membrane may be generally more resistant to control of glucose transport is suggested by failure or reduced ability of insulin, vitamin K5, and hydrogen peroxide to stimulate this function in older cells.