Relationship between the tissue level of cyclic AMP and the fat cell size of human adipose tissue. The relationship between mean fat cell size, maximal tissue cyclic AMP concentration, and glycerol release was investigated in human subcutaneous adipose tissue incubated in vitro with or without isoprenaline or noradrenaline added at maximal effective concentrations. Basal and stimulated glycerol release and cyclic AMP concentration were each related to the fat cell size. Whether or not the phosphodiesterase inhibitor theophylline was present in the incubation system, basal and noradrenaline-induced cyclic AMP levels were significantly correlated with the fat cell size. The noradrenaline-induced cyclic AMP levels resulted in twice as rapid glycerol release as could be expected from the basal ratio between glycerol release and cyclic AMP. Furthermore, both basal and noradrenaline-induced glycerol release in relation to the cyclic AMP levels were more rapid in enlarge fat cells. It is concluded that basal and catecholamine-induced production of cyclic AMP is related to the fat cell size and that a quantitative relationship exists between rate of lipolysis and maximal tissue levels of cyclic AMP in human adipose tissue. Basal and noradrenaline-induced lipolysis are probably regulated by different mechanisms and the lipolytic sensitivity to cyclic AMP seems increased in large fat cells.


Catabolism of the apoprotein of low density lipoproteins by the isolated perfused rat liver. Isolated rat livers were perfused for 4 hours in a recirculating system containing washed rat erythrocytes. Biologically screened, radioiodinated low density lipoproteins (1.030 < d < 1.055 g/ml) were added to the perfusate with different amounts of whole serum to supply unlabeled rat low density lipoproteins. Apolipoprotein B contained 90% of the bound (131)I, other apolipoproteins contained 4%, and lipids contained the remainder. The fraction of apolipoprotein mass degraded during the perfusion was quantified by the linear increment of non-protein-bound radioiodine in the perfusate, corrected for the increment observed during recirculation of the perfusate in the absence of a liver. The fractional catabolic rate ranged from 0.3 to 1.7%/hr in seven experiments and was inversely related to the size of perfusate pool of low density apolipoprotein. The catabolic rate of low density apolipoprotein (fractional catabolic rate x pool size) in four livers, in which the concentration of rat low density lipoproteins was 50-100% of that present in intact rats, was 5.3 +/- 2.7 micro g hr(-1) (mean +/- SD). Similar results were obtained with human low density lipoproteins. These rates were compared with catabolic rates for the apoprotein of rat low density lipoproteins in intact animals. Fractional catabolic rate in vivo, obtained by multi-compartmental analysis of the disappearance curve of (131)I-labeled low density apolipoprotein from blood plasma, was 15.2 +/- 3.1% hr(-1) (mean +/- SD). Total catabolic rate in vivo (fractional catabolic rate x intravascular pool of low density apolipoprotein) was 76 +/- 14 micro g hr(-1) (mean +/- SD). The results suggest that only a small fraction of low density apolipoprotein mass in rats is degraded by the liver.


Uptake and degradation of high density lipoprotein: comparison of fibroblasts from normal subjects and from homozygous familial hypercholesterolemic subjects. Fibroblasts cultured from the skin of subjects with homozygous familial hyperlipoproteinemia (HFH) internalize and degrade low density lipoproteins at a much lower rate than do fibroblasts from normal subjects. Evidence has been presented that this reflects the absence from such mutant cells of specialized binding sites with high affinity for low density lipoproteins. The specificity of this membrane defect in familial hypercholesterolemia is further supported by the present studies comparing the metabolism of low density lipoproteins (LDL) and high density lipoproteins (HDL) in normal fibroblasts and in fibroblasts from HFH patients. The surface binding (trypsin-releasable (125)I) of (125)I-labeled LDL by HFH cells was approximately 30% of that by normal cells at a concentration of 5 micro g LDL protein per ml. At the same concentration the internalization (cell-associated (125)I after trypsinization) and degradation (trichloroacetic acid-soluble non-iodide (125)I) of (125)I-labeled LDL were less than 10% of the values obtained with normal cells. In contrast, the binding of (125)I-labeled HDL to HFH cells was actually somewhat greater than that to normal cells. Despite this, the internalization and degradation of (125)I-labeled HDL by HFH cells averaged only 70% of that by normal cells. [(3)H]- or [(14)C]Sucrose uptake, a measure of fluid uptake by pinocytosis, was similar in normal and HFH fibroblasts. These findings are consistent with the proposal that fibroblasts from subjects with HFH lack high-affinity receptors for LDL. These receptors do not play a significant role in HDL binding and uptake. Instead, as previously proposed, HDL appears to bind randomly on the cell surface and its internalization is not facilitated by the specific mechanism that internalizes LDL. The small but significant abnormalities in HDL binding and internalization, however, suggest that there may be additional primary or secondary abnormalities of membrane structure and function in HFH cells. Finally, the observed overall rate of uptake of LDL (that internalized plus that degraded) by HFH fibroblasts was considerably greater than that expected from fluid endocytosis alone. This implies that adsorptive endocytosis, associated with binding to low-affinity sites on the cell surface, may play a significant role in LDL degradation by HFH cells, even though it does not regulate endogenous cholesterol synthesis in these cells.


A comparison of two methods to investigate the metabolism of human apolipoproteins A-I and and A-II. Two methods are compared for measuring the kinetic parameters of apolipoprotein A-I and A-II metabolism in human plasma. In the first, high density lipoprotein apoproteins were radioiodinated in situ in the lipoprotein particle (endogenous apoprotein labeling) while in the second, individually labeled apolipoprotein A-I or A-II was incorporated into the particle by in vitro incubation (exogenous apoprotein labeling). The catabolic clearance rate of exogenously labeled apolipoprotein A-I was consistently faster than that of endogenous apolipoprotein A-I. Conversely, endogenously and exogenously labeled apolipoprotein A-II were catabolized at identical rates. The fractional plasma clearance rates of endogenous apolipoproteins A-I and A-II were the same.


A study of the effect of environmental CO2 on experimental Wilm's tumor. The effects of exposure of experimental murine Wilm's tumor to increased atmospheric concentrations of carbon dioxide were studied. Local tumor growth was not affected by concentrations of 76% and 55% of carbon dioxide applied for 10 and 30 minutes. There was no difference in survival of animals and weight of the tumor between the control and the experimental groups. Ten per cent of the animals treated with concentrations of 55% and 76% carbon dioxide developed metastases in contrast to 30% of metastatic growth detected in the control group. This observation, although without statistically significant difference, may lead toward further study of the carbon dioxide effect in different tumor models. Additional tumor models and different animals species will be studied.


Stimulation of erythropoiesis in mice by adenosine 3',5'-monophosphate and prostaglandin E1. Adenosine 3',5'-monophosphate, 2 to 16 mg, and prostaglandin E1, 0.05 to 0.10 mg, stimulate erythropoiesis in ex-hypoxic, polycythemic mice. The stimulation can be prevented by the simultaneous administration of an antiserum to erythropoietin. India ink penetration shows that at these doses blood circulation is blocked in the kidney. Tissue hypoxia, especially in the kidney, causes production of erythropietin. The strong effect on the kidney circulation suggests that the major part of the stimulation of erythropoiesis by these agents is through their effect on renal circulation.


Sources of variation in the output of locust spiracular motoneurones receiving common synaptic driving. 1. The closer muscles of the left and the right spiracles of a thoracic segment are both innervated by two motoneurones, which spike in a variety of patterns during expiration. This paper seeks to explain the origin of these patterns. 2. No direct coupling between the two motoneurones is revealed. In an isolated thoracic ganglion both motoneurones spike at different frequencies with no tendency for their spikes to become synchronized. 3. The two closer motoneurones in one segment receive common, patterned depolarizing synaptic potentials during expiration caused by interneurones relaying information from the metathoracic ganglion. 4. The closer motoneurones of all the thoracic segments receive the same pattern of synaptic potentials from these interneurones. Despite this, the spiracles of one segment may remain shut while those on other segments continue to open and close rhythmically. 5. The interplay between common synaptic driving, the threshold of the motoneurones for spike initiation, and the tendency for a motoneurone to spike at a particular frequency even in the absence of interneuronal driving, explains the various patterns of spikes during expiration. Common synaptic driving imposes the same basic pattern of commands on all the motoneurones, but the individual motoneurones determine the final pattern of motor spikes. 6. To be effective in producing a patterned output, an input pattern must operate within narrow limits on either side of the threshold of the motoneurone. If the depolarization is too large, a high frequency of unpatterned spikes will result; if too small, then either there will be no output or a low frequency of spikes will result whose patterning will be affected by other inputs.


Effects of epinephrine on branchial non-electrolyte permeability in rainbow trout. Using isolated heads perfused at constant pressure, at rates close to those occurring in vivo, the permeability of the gills of the trout Salmo gairdneri to a range of solutes was measured. Under epinephrine-free conditions, butanol and water showed similar high branchial permeability coefficients. Urea, inulin and dextrans (mol. wt 3000 and 20 000) were 7-12 times less permeant, and mannitol 60-70 times less permeant than water or butanol. Epinephrine, at 10(-6) M, greatly increased the permeability of the gills to the small hydrophilic molecules, water and urea, and to the lipophilic substance, butanol, but did not affect the penetration of the large hydrophilic solutes, mannitol, inulin and dextrans. In the presence of 10(-6) M propanolol, a beta-blocker, epinephrine had no effect on the permeation of any of the test substances except that the permeability to urea decreased somewhat. The results suggest that epinephrine increases the permeability of the membranes of the branchial cells but does not affect the permeation of substances that cross the gill walls by paracellular routes or via an intracellular 'bulk-transport' mechanism. Such an action would be expected to increase the branchial transfer of oxygen.


Increased superoxide anion production by immunologically activated and chemically elicited macrophages. We studied the capacity of cultured mouse peritoneal macrophages to generate superoxide anion (O(2-)), the initial product of conversion of oxygen to microbicidal species, during phagocytosis of opsonized zymosan or upon contact with the membrane-active agent phorbel myristate acetate (PMA). Macrophages from mice infected with Bacille Calmette-Guerin (BCG) or injected intraperitoneally with thioglycollate broth or endotoxin, released up to 12 times more O(2-) than did resident peritoneal macrophages, depending upon the cell type and whether the stimulus was zymosan or PMA. There was little if any O(2-) release from resting (unstimulated) macrophages. The density of cells on culture dishes was an important variable since crowding of the dish markedly reduced the efficiency of O(2-) production. The enhanced O(2-) release of chemically elicited and infection-activated macrophages was noted after stimulation with a wide range of concentrations of PMA and zymosan, at all time points studied (up to 120 min), and with cells maintained for 140 rain to 16 days in culture. The O(2-) response of resident cells improved twofold to zymosan and ninefold to PMA during the first 3 days in culture. The capacity to release O~ appears to be limited to actively phagocytic cell types: murine macrophage-like tumor lines and cultured human monocytes released O(2-) when stimulated by PMA or zymosan, fibroblast and endothelial lines and embryo-derived cells did not. Activity of superoxide dismutase, which removes O(2-), was not detectable in culture supernates of any cell type, and thus, differences in detectable O(2-) could not be attributed to variations in the release of this enzyme. We conclude that the phagocytosis- associated respiratory burst is significantly enhanced in mononuclear phagocytes obtained ai~r chemical inflammation or BCG infection. Increased capacity to generate O(2-) and other oxygen radicals during phagocytosis could contribute to the improved microbicidal and tumoricidal activity of activated macrophages.


Restricted expression of ecotropic virus by thymocytes of leukemia-resistant (AKR X NZB)F1 mice. AKR mice, which produce high titers of ecotropic virus, were crossed with NZB mice, which produce titers of xenotropic virus. Spleen, marrow, and lymph node cells of the F1 hybrid produced high titers of ecotropic and xenotropic viruses. However, expression of ecotropic virus by both thymus cells and peripheral T cells of the F1 was severely restricted. Despite simultaneous expression of ecotorpic and xenotropic viruses in F1 spleens, lymph nodes, and marrows evidence for recombinant viruses was not found. Such viruses were also undetectable in the F1 thymuses. The results indicate that a cellular mechanism, present in AKR thymus but lacking in the F1 influences virus expression and the formation of recombinant viruses. This may account for the low incidence of leukemia in the F1 hybrid.


Virus-induced atherosclerosis.

Virus-induced atherosclerosis. Of four groups of chickens, two (groups I and II) were infected with MDV and two were not (groups III and IV). Groups I and III were fed diets low in lipid, and groups II and IV were fed cholesterol-supplemented diets. Striking grossly visible atherosclerotic lesions were seen in large coronary arteries, aortas, and major aortic branches of infected normocholesterolemic and hypercholesterolemic chickens (groups I and II). In contrast, grossly visible atherosclerotic lesions were not seen in uninfected normocholesterolemic chickens (group III), nor in uninfected hypercholesterolemic chickens (group IV). Microscopically, arterial changes in the infected animals were characterized by occlusive fibromuscular intimal thickening which formed fibrous caps overlying areas of atheromatous change. This change closely resembled chronic atherosclerosis in man. These results may have important bearing on our understanding of the etiology and pathogenesis of human arteriosclerosis since there is widespread and persistent infection of human populations with up to five different herpes-viruses.


Patterned response to odor in single neurones of goldfish olfactory bulb: influence of odor quality and other stimulus parameters. Responses of 75 single units in the goldfish olfactory bulb were analyzed in detail for their relationship to the time-course of the change in odor concentration during each odor stimulus. Odor stimuli were controlled for rise time, duration, and peak concentration by an apparatus developed for the purpose. This apparatus enabled aqueous odor stimuli to be interposed into a constant water stream without changes in flow rate. The time-course of the concentration change within the olfactory sac was inferred from conductivity measurements at the incurrent and excurrent nostrils. Temporal patterns of firing rate elicited by stimuli with relatively slow rising and falling phases could be quite complex combinations of excitation and suppression. Different temporal patterns were produced by different substances at a single concentration in most units. Statistical measures of the temporal pattern of response for a small number of cells at a given concentration were more characteristic of the stimulus substance than any of three measures of magnitude of response. The temporal patterns change when the peak concentration, duration, and rise time of the stimuli are varied. The nature of these changes suggests that the different patterns are due primarily to the combined influence of two factors: (a) a stimulus whose concentration varies over time and (b) a relationship between concentration and impulse frequency which varies from unit to unit. Some units produce patterns suggestive of influence by neural events of long time constant. The importance of temporal patterns in odor quality and odor intensity coding is discussed.


Enhanced parainfluenza I (6/94) virus detection in latently infected human brain cell cultures by treatment with cytochalasin D and dimethyl sulfoxide. The ability of cytochalasin D (CD) and dimethyl sulfoxide (DMSO) to enhance parainfluenza I (6/94) virus replication was studied in various cell culture systems. Treatment of CV1 cells with CD (1 microgram/ml) dissolved in DMSO prior to primary 6/94 virus exposure at 10(0)--10(5) multiplicities of infection did not substantially enhance virus replication. However, there was a transient increase in cell associated virus one day after infection of DMSO-treated cultures. CD treatment of cultures of human brain cells latently infected with 6/94 virus (LIHB cells) did not enhance 6/94 virus detection. Cocultivation of CV1 cells with CD-treated LIHB cell cultures, and cocultivation of LIHB cell cultures with CD-treated CV1 cells, resulted in the production of both cell-associated and cell-free 6/94 virus three and five days after cocultivation. No virus was detected after similar cocultivation of untreated LIHB cell cultures with untreated CV1 cells. The usefulness of CD-DMSO treatment in the rescue of virus from 6/94 LIHB cell cultures appears limited to a cocultivation system. The use of these techniques to enhance virus rescue from human tissues suspected of harboring latent viral genomes is discussed.


In vivo responses of paired giant mechanoreceptor neurons in Aplysia abdominal ganglion. Two neurons with cell bodies symmetrically located in the abdominal ganglion and giant axons in the left (L1) and right (R1) pleurovisceral connectives of Aplysia californica were examined in vivo and in vitro. Direct stimulation of R1 and L1 in the intact animal does not elicit any observable behavior, suggesting that they are neither motoneurons nor command neurons. These cells respond in vivo to sudden onset mechanical stimulation of widespread regions of the body. R1 and L1 spikes are initiated in at least three different loci: (1) the peripheral axon in the foot, (2) the neuropil of the pleural and/or pedal ganglion, and (3) the neuropil of the abdominal ganglion. Furthermore, R1 and L1 probably have two different mechanisms for spike initiation: (1) sensory (foot), and (2) synaptic (abdominal and/or head ganglia). The different loci for spike initiation account for the bidirectional conduction of R1 and L1 spikes. As sensory (mechanoreceptor) neurons, R1 and L1 have peripheral axons in the ipsilateral posterior pedal nerve, show low threshold responses to stimulation of the ipsilateral posterior foot, they are rapidly adapting their responses do not decrease with repetition, and they are not blocked by high Mg++/low Ca++ solutions. As synaptically-driven neurons, R1 and L1 have widespread bilateral responsiveness, their responses decrease with repetition and their inputs are blocked with high Mg++/low Ca++ solutions. These neurons integrate sensory and synaptic inputs and conduct bidirectionally, however, their output connections must be specified before their behavioral function can be understood.


Reflex arc of the first component of the human blink reflex: a single motoneurone study. Latency variation of consecutive responses of single orbicularis oculi motoneurones in the first component of the electrically elicited blink reflex was 2.6 times larger than that of the H reflex of the soleus muscle fibres. Some motoneurones showed bimodal latency distribution or double responses or both at an interval of about 4-5 ms. Typical first component responses were sometimes obtained by contralateral stimulation. Central conduction time in the brainstem, estimated for 15 motoneurones, was between 2.4 and 6.6 ms. It is concluded that the first component of the blink reflex is conducted through an oligosynaptic arc including one or more interneurones.


Radiculopathy as a complication of ankylosing spondylitis. A cauda equina syndrome in association with ankylosing spondylitis is well recorded. We report the occurrence of an upper limb radiculopathy which progressed to a cauda equina syndrome in association with ankylosing spondylitis.


Surface topography of normal and neoplastic human anterior pituitary cells maintained in vitro. Pituitary tissues were obtained from 25 patients who underwent surgery for excision of pituitary macroadenomas, selective excision of microadenomas, or removal of a normal gland for palliation of metastatic cancer. Cells thus obtained were maintained in vitro for varying intervals, fixed, and examined by light (phase contrast), microscopy, transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Previous SEM reports indicate that surface topography of in vitro neoplastic cells displays features that may correlate with neoplastic behavior. Cultured normal and pituitary tumor cells did not display these surface differences, with one exception, a prolactin-secreting microadenoma. Characteristic patterns for the cell populations were identified. Certain cell types appeared in all the cultures: 1) large and small granule-containing cells; 2) flat and irregular cells; 31 spindle-shaped cells; and 4) spherical, irregularly surfaced cells. In one case of an endocrine-inactive juvenile pituitary chromophobe adenoma, unique cells were observed. Surface topography did not appear to be of predictive value in determining the neoplastic character of pituitary tumors.


Intraneural hematoma of the sciatic nerve. Case report. A case of an encapsulated organizing hematoma of the sciatic nerve is reported. Hemorrhage-induced neuropathy as a complication of anticoagulant therapy, leukemia, hemophilia, and other bleeding diatheses has been frequently reported. While trauma is the most common etiology of a hemorrhagic neuropathy, actual hematoma formation beneath the epineurium is very rare.


Studies on carbamoylphosphate synthetase in livers of rats fed a protein free diet supplemented with methionine and threonine and injected with N-acetylglutamic acid. Supplementation of methionine and threonine to a protein free diet reduces urinary nitrogen excretion in rats, and it has been assumed that N-acetylglutamic acid may have a regulatory role in urea synthesis. The present paper reports the effect of supplementation of methionine and threonine on the levels of N-acetylglutamic acid and of carbamoylphosphate synthetase (EC 2.7.2.5) in liver. In spite of a significant decrease in urinary urea when methionine and threonine were supplemented to the protein free diet, the activity of carbamoylphosphate synthetase was not reduced. On the other hand, following injection of glutamic acid, N-acetylglutamic acid or N-acetylglutamine, the excretion of urinary urea of rats fed the protein free diet supplemented with methionine and threonine was significantly reduced whereas the level of carbamoylphosphate synthetase was the same in each group. Furthermore, the concentrations of N-acetylglutamic acid in liver of rats fed the protein free diet or the protein free diet supplemented with methionine and threonine were the same. Therefore, these results indicated that, when methionine and threonine were supplemented to the protein free diet, the changes in the levels of N-acetylglutamic acid and of carbamoylphosphate synthetase in liver does not contribute to the reduced body weight loss and urinary nitrogen excretion.


Differential response of calcium transport systems in laying hens to exogenous and endogenous changes in vitamin D status. Changes in intestinal calcium absorption, calcium deposition into egg shell, and intestinal, renal and uterine calcium-binding protein (CaBP) in laying hens were related to changes in 25 hydroxycholecalciferol-1-hydroxylase activity (1-hydroxylase), or to the supplementation of 1alpha-hydroxycholecalciferol (1alpha-OH-CC). The onset of egg production resulted in an increased kidney 1-hydroxylase activity and intestinal and uterine CaBP. Renal concentrations of CaBP remained unchanged. Intestinal calcium and phosphorus absorption, during the period of egg shell formation, and duodenal calcium-binding protein (CaBP), were higher in 1alpha-OH-CC-fed than in cholecalciferol-fed hens. Renal or uterine CaBP and calcium deposition into the egg shell did not fluctuate with the vitamin D source or concentration. 1alpha-OH-CC injection into non-laying Nicarbazinfed hens resulted in an increase in intestinal but not renal or uterine CaBP concentrations. It is suggested that (a) CaBP in various organs responded independently to the same stimuli; and (b) calcium deposition into egg shell and uterine CaBP level are not related to kidney 1-hydroxylase activity or concentration of 1,25 dihydroxycholecalciferol.


Hydrogen cyanide poisoning: treatment with cobalt EDTA. Three case reports are presented of employees who suffered varying degrees of exposure to hydrogen cyanide and their subsequent clinical courses following treatment with cobalt EDTA. A review of treatment modalities for CN- toxicity is given. It is concluded that, because of the degree of patient symptomatology associated from the use of cobalt EDTA, this therapy be reserved only for patients with the most severe degress of exposure to CN(-), and that in all other cases combined sodium nitrite and sodium thiosulphate therapy should be employed.


The course of five-year survivors of cancer in childhood. A follow-up study was made of two independent series of 1,807 and 425 children who were alive five years after diagnosis of a malignant neoplasm. Over the subsequent 20 years (5 to 24 years after initial diagnosis), actuarial survival rates for the cohorts were 83% and 79%, respectively, compared to 97% for matched control subjects in the general population (P less than 0.01). Cure was achieved in a large majority of the 5-year relapse-free survivors of cancer in this study.


Premature craniosynostosis: A common complication of juvenile thyrotoxicosis. Cranial vault suture opacification (apparent closure) and bone age were evaluated roentgenographically in ten children with thyrotoxicosis. The bone age was advanced greater than 2 SD in only one. In comparison to 96 control children of similar age, craniosynostosis was present in each of the patients with thyrotoxicosis. Children with advanced bone age, nine due to virilizing adrenal hyperplasia and three with precocious puberty, had normal radiographic patterns of cranial suture closure. Thyrotoxic premature craniosynostosis did not interfere with continued head circumference growth nor did it result in clinical or radiographic evidence of increased intracranial pressure. We conclude that premature craniosynostosis appears to be a common feature of juvenile thyrotoxicosis. Investigation of the possible long-term adverse effects of this entity on central nervous system function is advocated.


Congenital lipodystrophy: An endocrine study in three siblings. I. Disorders of carbohydrate metabolism. Three siblings with congenital lipodystrophy were studied extensively for endocrine abnormalities. A severe disturbance in carbohydrate metabolism was observed. Plasma concentrations of glucagon and insulin were markedly elevated both in the basal state and in response to provocative stimuli. In addition, marked resistance to exogenous insulin and a diabetic oral glucose tolerance test were demonstrated. Lipid metabolism, GH, and ACTH secretion were normal...


Experience with "second-look" operations in pediatric solid tumors. "Second-look" operations were performed in 32 infants and children with initially unresectable or recurrent solid tumors treated with combination chemotherapy and/or irradiation. Tumors were resectable in 26 of 32 cases (81%). These procedures often yielded information affecting staging and treatment. Disease-free survival was achieved in 18 of 32 patients (56%). Mortality was related to progressive disease in seven cases and opportunistic infections due to immunosuppression in three. Four additional patients are alive with evidence of persistent tumor. Second-look procedures were beneficial in patients with Wilms' tumor previously operated upon by a flank approach and in children with bilateral tumors. These procedures were particularly useful in children with stage III localized neuroblastoma and cases of metastatic neuroblastoma that respond to chemotherapy. Second-look operations were also useful in selected cases of rhabdomyosarcoma, teratoma, and Ewing's sarcoma. These observations suggest that combination chemotherapy has increased the use of second-look operations in a variety of less favorable (e.g. initially unresectable or recurrent) pediatric solid tumors.


Treatment strategy for nodular renal blastema and nephroblastomatosis associated with Wilms' tumor. Nodular renal blastema and nephroblastomatosis were present in 8 of 118 patients (6.8%) with Wilms' tumor. Five of these 8 patients (63%) had bilateral Wilms' tumors. Two had hemihypertrophy. Preoperative renal angiograms were accurate in detecting these metanephric anomalies. The surgical approach consisted of removal of the most diseased kidney and biopsy for diffuse tumors and wedge resections for localized tumors for the remaining kidney. Postoperatively, radiation was administered when tumor extended outside the kidney. Chemotherapy consisted of vincristine and dactinomycin for 18 mo and adriamycin for 6 mo. This method of management resulted in tumor-free survival of these 8 patients for 1--44 mo (median 24 mo). Nodular renal blastema and nephroblastomatosis may possibly develop into Wilms' tumor. All of these three conditions respond to surgery, chemotherapy, and radiation. When a Wilms' tumor is encountered, it is better to explore and possibly biopsy the opposite kidney. There is a place for second-look laparotomy in this spectrum of congenital anomalies.


Tumor invasion of the upper inferior vena cava: the use of profound hypothermia and circulation arrest as a surgical adjunct. Two children presented with abdominal tumors invading the upper inferior vena cava. To facilitate removal of the tumors, cardiopulmonary bypass with profound hypothermia and circulation arrest were utilized. This technique simplifies tumor excision and is worthwhile even in children with advanced malignant disease.


The effect of guanidine on transmitter release in the ciliary ganglion of the chick. 1. The effect of guanidine hydrochloride on synaptic transmission at the chick ciliary ganglion was studied. 2. In bathing solution containing low calcium concentrations, guanidine produced a dose-dependent increase in the number of quanta released by the presynaptic action potential. No post-synaptic effects were observed. 3. Intracellular presynaptic recordings and monitoring of the presynaptic action potential post-synaptically via the coupling potential showed that the action of guanidine was independent of any alterations in the presynaptic spike.


Oxygen consumption and neonatal sleep states. 1. In thirty full-term infants in the first week of life, nursed in a constant volume, closed-circuit metabolism chamber in a neutral thermal environment (31.5-33.5 degrees C), measurements were made of oxygen consumption ( V(O2)) during periods of rapid eye movement (REM) sleep and non-rapid eye movement (NREM) sleep.2. The mean V(O2) during REM sleep was 5.97 ml. kg(-1). min(-1). In NREM sleep the mean V(O2) was 5.72 ml. kg(-1). min(-1). This difference was significant (paired t test P < 0.05).3. When the direction of sleep state change was taken into account the difference in V(O2) between the two states was much less when REM sleep preceded NREM than when the change was in the opposite direction. In nineteen infants in whom the change was from REM to NREM the difference in V(O2) (6.18 and 6.03 ml. kg(-1). min(-1)) was not significant (P > 0.05). The mean difference when the sleep state change was from NREM to REM was significant (P < 0.01), the values being 5.54 and 5.81 ml. kg(-1). min(-1) respectively.4. In the NREM state, a gradual diminution of V(O2) with time was consistently found. This was not the case in REM sleep.5. In twelve infants studied in a cool environment (29 +/- 0.5 degrees C) V(O2) during REM sleep was 7.77, and during NREM sleep it was 6.58 ml. kg(-1). min(-1), (P < 0.001). Thus even the maximum difference found in a neutral thermal environment of 6.6% was significantly increased to 14.9% (P < 0.01) with mild thermal stress.6. No consistent changes in V(O2) with time were found in either REM or NREM sleep in twelve infants studied in a cool environment, in contrast to the findings in thermal neutrality


Quelling of spontaneous transmitter release by nerve impulses in low extracellular calcium solutions. 1. The effect of nerve stimulation on spontaneous transmitter release was studied at the frog neuromuscular synapse which was bathed in a solution containing very low extracellular calcium concentration. Conventional methods for intracellular and extracellular recording were used and the pattern of quantal liberation following the nerve stimulus was determined. 2. Stimulation of the motor nerve (at rates between 0.09 and 2Hz) caused a reduction in the frequency of the miniature e.p.p.s in comparison to the prestimulation values. 3. The mean distribution of the time of occurrence of the miniature e.p.p.s during the interstimulus period showed periodic oscillations. 4. The quelling effect of nerve stimulation on transmitter release is explained by the hypothesis that a low [Ca]o a reversed electrochemical gradient for calcium occurs and nerve stimulation causes an increased calcium conductance leading to calcium efflux which in turn temporarily reduces [Ca]i and transmitter release.


The role of calcium ions in tetanic and post-tetanic increase of miniature end-plate potential frequency. 1. The role of Ca ions in transmitter release changes, during and after high frequency stimulation of the motor nerve (10--100 Hz), was examined at the frog neuromuscular junction. 2. The stimulation-induced changes in miniature end-plate potential frequency (f) resembled the changes in end-plate potential amplitude recently described by Magleby and Zengel (1975, 1976). 3. The effects of tetanic stimulation on f under inward electrochemical gradient for Ca ions were compared with those under reversed gradient and four differences were found: (a) The increase in f during the tetanus under reversed Ca gradient conditions is much smaller than with an inward Ca gradient. (b) The increase in f under reversed Ca gradient is preceded by a small decrease in f, whereas with an inward Ca gradient an immediate increase in f is observed. (c) After the termination of the tetanus with a reversed Ca gradient, there is a further increase in f, compared to a decrease with an inward Ca gradient. (d) The augmentation phase of post-tetanic potentiation was practically abolished. 4. The experimental results are explained by assuming that high frequency nerve stimulation causes an increase in transmitter release by at least two distinct processes: influx of Ca ions through the presynaptic membrane and release of Ca ions from intracellular stores. It is suggested that Na ions couple nerve activity to intracellular release of Ca.


Clumping and oscillations in evoked transmitter release at the frog neuromuscular junction. 1. Time series analysis of evoked transmitter release was performed at the frog neuromuscular synapse. 2. Clumping of end-plate potentials with similar amplitude was found in the time domain. 3. At low quantal contents periodic oscillations were observed with a period of 14 sec. 4. Clumping and oscillations are phenomena of presynaptic origin. 5. The results are explained on the hypothesis that periodic fluctuations occur in Ca concentration inside the presynaptic nerve terminal.


Oxytocin release following osmotic activation of oxytocin neurones in the paraventricular and supraoptic nuclei. 1. Recordings were made from a total of 35 antidromically identified neurones in the paraventricular (PV) and supraoptic (SO) nuclei of urethane-anaesthetized lactating rats. During recording plasma osmotic pressure was raised by 12 m-osmole/kg by injection of hypertonic solutions of NaCl, LiCl, or mannitol.2. Nine PV neurones (mean firing rate 4.2 +/- 1.0 (S.E.) spikes/sec) were classified as oxytocin cells because they gave a burst of activity before reflex milk-ejections. None of these showed a bursting (phasic) firing pattern. Ten PV neurones (mean firing rate 1.8 +/- 0.2 spikes/sec) fired phasically either before or after injection of hypertonic NaCl and were classified as vasopressin cells. The remaining six PV cells (mean firing rate 1.6 +/- 0.9 spikes/sec) showed no bursts of firing related to milk ejection and did not fire phasically.3. Increasing plasma osmotic pressure by injection of hypertonic NaCl increased the mean firing rate of PV oxytocin cells to 7.0 +/- 1.0 spikes/sec. Vasopressin cells in the PV nucleus were much less responsive and the mean firing rate after injection was 2.9 +/- 0.4 spikes/sec. The third group of PV neurones was unresponsive.4. Plasma oxytocin concentration (determined by radioimmunoassay) increased from 2.1 +/- 0.3 muu./ml. in the control period to 10.9 +/- 2.8 muu./ml. 30 min after I.P. injection of 1 ml. 1.5 M-NaCl and to 14.8 +/- 2.8 muu./ml. following injection of a second 1 ml. 1.5 M-NaCl.5. The responses of oxytocin and vasopressin neurones in the SO nucleus to an increase in plasma osmotic pressure following injections of hypertonic solutions of LiCl or mannitol were similar to those observed when plasma osmotic pressure was raised by NaCl.6. It may be concluded that both oxytocin and vasopressin cells in the neurohypophysical system are responsive to the osmotic pressure of the blood plasma rather than to Na(+) or Cl(-) concentration, that osmotic activation of oxytocin cells releases sufficient oxytocin to increase its plasma concentration, and that there may be a functional difference between the SO and PV nuclei.


Control of calcium channels by membrane receptors in the rat parotid gland. 1. The mechanism of action of agonists that stimulate K release from the parotid gland was investigated by monitoring efflux of 86Rb from rat parotid slices. 2. As in previous studies, the 86Rb release was increased by agonists in a biphasic manner. An early, transient phase occurred that was independent of extracellular Ca. This was followed by a sustained (or slowly falling phase) that required extracellular Ca. 3. The agonists were investigated as to their additivity and to their sensitivity to inhibition by a number of putative Ca-antagonists. 4. When carbachol, epinephrine and Substance P were employed in supramaximal concentrations, no combination of agonists produced a summated 86Rb release response, despite the fact that the Ca concentration was submaximal. 5. The sustained phase of 86Rb release, which is dependent on extracellular Ca, was blocked by La, Ni, Co and neomycin; the transient phase was unaffected by these agents. 6. The local anaesthetics tetracaine and procaine inhibited both the transient and sustained phases of the responses to carbachol and phenylephrine; responses to Substance P and to the divalent cationophore, A23187, were largely refractory to this effect. 7. These results support the contention that, in the parotid, muscarinic, alpha-adrenergic and peptide receptors regulate the same Ca influx sites. 8. Also, these results suggest that La, Ni, Co and neomycin appear to antagonize the action of Ca by impeding inward movement of the cation through activated Ca influx sites. 9. Finally, the local anesthetics appear to inhibit, and therefore, serve to define, a transduction step between receptor occupation and channel activation. 10. In the case of the Substance P mechanism, it appears that the transduction mechanism must be qualitatively different to that for the muscarinic or alpha-adrenergic mechanisms.


Temperature-sensitive aspects of evoked and spontaneous transmitter release at the frog neuromuscular junction. 1. The temperature dependence of presynaptic processes involved in neuromuscular transmission was studied by rapidly increasing the temperature of cooled frog neuromuscular junctions by 4--10 degrees C using pulses from a neodymium laser. The temperature elevation was complete within 0.5 msec, and decayed back to control levels with a time constant of about 7--8 sec. 2. Temperature jumps completed before nerve stimulation increased the quantal content and decreased the latency of the end-plate potential (e.p.p.). The Q10 for e.p.p. quantal content in low [Ca2+] Ringer averaged about 3.9 over the range 1--18 degrees C. 3. Temperature jumps occurring during the synaptic delay (the interval between the presynaptic action potential and the onset of the e.p.p.) also increased the quantal content and decreased the latency of the e.p.p. These effects diminished as the onset of the temperature jump was moved closer to the expected onset of the e.p.p. Temperature jumps applied after the onset of the e.p.p. immediately accelerated the time course of the e.p.p. but did not significantly alter quantal content. These results demonstrate that the magnitude and timing of evoked release are influenced by temperature-sensitive processes that operate both during and shortly after the presynaptic nerve action potential, but are largely complete before the onset of release. 4. Temperature jumps were applied at various times during the interval between two nerve stimuli. The amplitude of the second e.p.p. decreased as the temperature jump was moved earlier in the interstimulus interval, suggesting that the rise in temperature following the first nerve stimulus accelerates the decay of facilitation. When the temperature jump was moved from 10 msec after to 10 msec before the onset of the first e.p.p., the amplitude of the second e.p.p. either decreased or showed no change. The fact that the second e.p.p. did not increase suggests that the temperature-sensitive processes that increase the quantal content of the conditioning e.p.p. do not greatly increase the facilitation following that e.p.p. 5. Temperature jumps immediately accelerated the time course of spontaneous miniature end-plate potentials (m.e.p.p.s) and increased their frequency. Experiments using slow temperature changes revealed that the Q10 for m.e.p.p. frequency in normal Ringer is about 10 over the range 10--20 degrees C. M.e.p.p. frequency was much less sensitive to temperature changes below about 10 degrees C. When the nerve terminal was depolarized by 20 mM-K+ in the presence of Ca2+, the Q10 for the rate of spontaneous release over the range 10--20 degrees C decreased to about 4, similar to the Q10 for e.p.p. quantal content. In the absence of extracellular Ca2+ the Q10 for m.e.p.p. frequency in 20 mM-K+ remained near 10. 6. The marked difference in Q10S for spontaneous transmitter release under different experimental conditions suggests that not all transmitter release uses identical mechanisms...


Synaptic delay in the heart: an ionophoretic study. 1. Neurotransmitters were applied ionophoretically to spontaneously beating clusters of ventricular muscle cells cultured from neonatal rats. 2. Acetylcholine or its analogue carbachol produced hyperpolarization and decreased the rate of spontaneous beating. These responses had minimum latencies of about 250 msec and total durations of 6-12 sec. 3. Noradrenaline, adrenaline or isoprenaline increased the rate of spontaneous beating. The minimum latency for this effect was 3-6 sec. Following a single brief pulse the rate remained elevated for 2 min or more. 4. Chronotropic responses of intact atria from adult rats to stimulation of the autonomic nerves were of similar time course to responses of the cultured muscle cells. 5. Calculations based on the theory of diffusion showed that access of drugs to their receptors could not be rate-limiting for the observed responses, unless a diffusion barrier of rather special properties was postulated. A number of other explanations for the long latencies have been ruled out; these are most likely to be due to some physical or chemical process occurring in or under the cell membrane. 6. Attempts to mimic responses to catecholamines by intracellular application of cyclic AMP were unsuccessful, perhaps because the release of nucleotide from the pipettes was insufficient. A theoretical treatment suggests that ionophoretic efflux of anions might be greatly diminished by the opposing electro-osmotic flux.


Synaptic transmission to the horizontal cells in the retina of the larval tiger salamander. 1. The receptive field diameter for most horizontal cells far exceeds the lateral spread of processes for any cell. Therefore horizontal cells probably receive synaptic input from neighbours as well as from the photoreceptors. The electrical effects of these two synaptic inputs were studied. 2. We have characterized the electrical properties of the horizontal cell inputs by determining the current-voltage curves in dark and light. These curves were compared with those obtained in the presence of Co2+ or Mg2 was nearly identical to the curve in the light. 4. The putative transmitter substances glutamate, aspartate and GABA depolarized the cells by increasing conductance. Current-voltage curves measured in the presence of these substances intersected the dark and light curves at +50 mV, the same level at which the dark and light curves intersect. 5. The light response of cells uith broad receptive fields, between 1.0 and 2.0 mm, showed little or no change in conductance associated with the light response. The input resistance was near 20 Momega, and the current-voltage curves intersected at an extrapolated potential level near 200 mV. 6. In the presence of ACh, electrical properties of the broad field cells reverted to those of the narrow field cells: the receptive field was reduced to 0.5 mm, the imput resistance increased, and the current-voltage curves intersected near +50 mV. Thus ACh appeared to interrupt synaptic input from neighbouring horizontal cells. 7. The results confirm the suggestion that horizontal cells receive a tonic excitatory input from the photoreceptors which is decreased by light. They show that horizontal cells receive an additional input from their neighbours, not associated with a measurable conductance change. The input from neighbours is selectively interrupted by ACh, but the nature of this synapse and of the cholinergic action is not known.


Mechanisms of post-synaptic excitation in amphibian motoneurones. 1. Post-synaptic excitation produced in motoneurones of the isolated perfused frog spinal cord by different monosynaptic inputs and by ionophoretically applied glutamate was analysed with intracellular recording technique. 2. Ca2+-deficient, high Mg2+ (5--20 mM) media or addition of Mn2+ (2mM) or Co2+ (5 mM) reversibly abolished chemically mediated e.p.s.p.s derived from medullary reticular formation, ventral and lateral columns, but not the short-latency, rapidly rising e.p.s.p.s derived from dorsal roots or muscle nerves, suggesting electric coupling between some primary afferents and spinal motoneurones. This conclusion is consistent with the dynamic properties of dorsal root e.p.s.p.s, their small sensitivity to cooling, and with results of correction of intracellular records made for contribution of extracellular field potential. E.p.s.p.s evoked by ventral root stimulation were also insensitive to Ca2+-lack and presence of 5--10 mM-Mg2+. 3. As the post-synaptic membrane was made more negative the amplitude of electrotonic dorsal root e.p.s.p.s was increased, and it was decreased by depolarizing currents. No reversal of the early part of the electrotonic e.p.s.p. was observed, although the presence of the local response would account for the occasional reversal of its later phase seen with depolarization. 4. When hyperpolarizing and depolarizing currents were applied to motoneurones in which chemically mediated e.p.s.p.s of the reticular cells, the ventral and lateral columns, were evoked, the actual reversal of the early part of e.p.s.p. was not observed, and there was no correlation between the sensitivity of the e.p.s.p.s to injected currents and their time course. The positive values of the extrapolated reversal potentials and the effects of changes in ionic content of perfusing media suggest that synaptically released transmitter triggers off the Na permeability of the subsynaptic membrane. 5. The amplitude of depolarization produced by ionophoretically applied glutamate depends non-linearly on membrane potential and the curvature of this dependence differs from that seen with chemically mediated s.p.s.p.s. The asymptotic nature of this relationship is explicable by a dependence of the membrane conductance change upon the membrane voltage. 6. The results of conductance measurements during the glutamate induced depolarization, the values of apparent reversal potentials and their dependence on external Na+ and K+ and internal Cl- is explicable by the opening post-synaptic channel gates for Na+ and closing post-synaptic channel gates for K+. 7. Chemical and electrical transmission in the amphibian cord is discussed in relation to recent anatomical findings.


Competitive antagonism by phentolamine of responses to biogenic amines and the transmitter at a neuroglandular junction. 1. A quantitative study has been made of phentolamine's inhibition of the electrical and secretory responses of the isolated salivary glands of Nauphoeta cinerea Olivier to nerve stimulation and bath applications of agonists. 2. The results suggested that phentolamine is a competitive antagonist having an affinity constant of about 1 micrometer-1 for the receptors for dopamine, noradrenaline, adrenaline and the neurotransmitter. 3. Phentolamine also inhibited responses to 5-hydroxytryptamine in a manner seemingly more complex than competitive antagonism. Attempts to estimate the affinity constant gave values of about 0.08 and 0.015 microgram-1 for inhibition of the secretory and electrical responses respectively. 4. This investigation showed that phentolamine discriminates between two kinds of receptor in this gland, one binding 5-hydroxytryptamine and the other combining with catecholamines and the neurotransmitter.


Light-activated hydrolysis of GTP and cyclic GMP in the rod outer segments. 1. The hydrolysis of guanosine triphosphate (GTP) and the consequent formation of guanosine diphosphate (GDP) and phosphate (P1) are activated by light in a suspension of broken retinal rods: the hydrolysis rate with GTP in the micrometer concentration range is 2.5-3.5 n-mole/min per mg of rhodopsin in the preparation. 2. The ionic composition of the medium suspending the rods is not critical: the hydrolysis is present in NaCl saline solution with MG2+ as well as in Tris-HC1 buffer solution, and with the chelating agent EDTA. 3. The ionic strength is critical: the effect is reduced when the broken rods are suspended in a low salt mannitol solution, and is altogether abolished when they are separated from the mannitol solution; it reappears when the mannitol solution is added again in the presence of salts. An element essential for the effect is thus reversibly released in the mannitol solution. No hydrolytic activity on GTP, however, is found in the mannitol soluble fraction. 4. The cyclic nucleotide phosphodiesterase is eluted from the rods in the mannitol solution, and is reaggregated to the rods in the presence of salts; once recombined with the rods, it can be activated by light. 5. The activation of the phosphodiesterase by light is present in the absence of added nucleotide triphosphates.


Adenosine-induced release of cyclic adenosine 3',5'-monophosphate from the left ventricle in the anaesthetized intact dog. 1. The influence of adenosine on the release of cyclic adenosine 3',5'-monophosphate (cyclic AMP) from the heart was examined in twenty-two anaesthetized intact dogs. The animals were pre-treated with propranolol and vagotomized. The rate of nucleotide release from the left ventricle was determined as the product of the left ventricular myocardial plasma flow and the coronary veno-arterial difference in plasma nucleotide concentration. 2. Adenosine was infused into the left ventricle during three successive 15 min periods at rates of 25, 50 and 100 n-mole/kg. min, respectively. The mean blood pressure in the ascending aorta was prevented from falling by inflating a balloon placed into the thoracic aorta. The measurements were performed before the infusion and at the 10th min of each infusion period. The values given are means +/- S.E. 3. During the 45 min infusion of adenosine at increasing rate, the cyclic AMP concentration in arterial plasma increased from 7.0 +/- 0.3 to 14.0 +/- 0.9 p-mole/ml. The nucleotide was released from the left ventricle at rates increasing from 48.2 +/- 7.1 to 206.5 +/- 56.2 p-mole/100 g. min while the left ventricular myocardial blood flow increased from 127 +/- 6 to 399 +/- 33 ml./100 g.min. The oxygen consumption of the left ventricle was not modified. 4. When adenosine was infused at a rate of 100 n-mol/kg.min, the thorax was opened and the apex of the heart and the left atrial appendage were removed for nucleotide assay. The cardiac cyclic AMP concentration did not differ from that observed in control dogs. 5. The results suggest that cyclic AMP is likely to be involved in the membrane and cellular events underlying the relaxant effect of adenosine on the coronary smooth muscle. The lack of change in cardiac cyclic AMP concentration, as determined by whole tissue extractions, is consistent with a study by others showing that, under normoxic conditions, cyclic AMP is released from a small, compartmentalized fraction of the cyclic AMP content of the heart. The elevation of the plasma nucleotide concentration could result from adenosine effects on various cell systems or organs, in addition to the observed release from the heart.


Gold-induced thrombocytopenia.

Gold-induced thrombocytopenia. Ten patients with rheumatoid arhritis treated with gold sodium thiomalate developed thrombocytopenia without bone marrow asplasia. There was no life-threatening blood loss, but petechiae, purpura, or echymoses were seen in eight patients. The serum gold levels monitored in one patient did not exceed levels seen in patients without thrombocytopenia. In three cases peripheral blood lymphocytes were cultured in the presence of gold and tritiated thymidine incorporation was significantly increased. Eight patients responded to steroid therapy, one patient to BAL and pencillamine, and one patient to vincristine.


Conformation of 2,9-dimethyl-3'-hydroxy-5-phenyl-6,7-genzomorphan and its relation to other analgetics and enkephalin. X-ray crystallographic data for 2,9-dimethyl-3'-hydroxy-5-phenyl-6,7-benzomorphan (I) as its p-bromobenzoyl ester are presented. The structure of I is compared with that of morphine, meperidine, alpha-allylprodine, methadone, and moramide as well as with a proposed structure of the enkephalins. A quantitative relationship is found between in vitro opiate receptor binding potency and in vivo analgesia for analgesics of diverse structure, including I. A new view of the analgetic pharmacophore is presented. Programs for the TI Programmable 59 calculator are described for conversion of X-ray crystallographic data to rectangular coordinates with reorientation of the molecule and for the calculation of torsion angles.


Aromatic esters of nonquaternary carbon-4 piperidinols as analgesics. Aromatic carboxylic esters of 1-methyl-4-piperidinol were prepared and evaluated for analgesic activity. In addition, aralkyl, alkyl, and cycloalkyl carboxylates of the 4-piperidinol system and 3,4-dimethoxybenzoates of isomeric piperidinols (24-26) were synthesized. The 3,4-dimethoxybenzoate 23 was nearly twice as active as codeine in the mouse hot-plate assay. In monkeys, 23 showed no morphine-like physical dependence liability, cis- and trans-1,3-dimethyl-4-piperidinol esters 24 and 25 showed no binding to the opiate receptor in rat brain homogenates. The 3- and 4-monosubstituted and the 3,4-disubstituted benzoate esters were examined for qualitative structure-activity relationships with respect to parameters Esc and pi. Various structural features of this series of compounds that may have an affinity for receptor binding sites are discussed.


Antiviral activity of some beta-diketones. 3. Aryl bis(beta-diketones). Antiherpetic activity. A series of bis(beta-diketones) was synthesized and tested in vitro for antiviral actitity against herpes simplex type 2. Two parameters which were studied in an effort to optimize activity were the nature of the aryl group and the length of the alkyl bridge. One of the more active compounds, 4,4'-[(1,4-phenylenedioxy)bis(6,1-hexanediyl)]-bis[3,5-heptanedione] (6), was evaluated more extensively and found to inhibit the cytopathic effect in tissue culture of herpes simplex virus type 1 as well as type 2. Compound 6 was evaluated in vivo topically against herpes simplex type 1 in experimentally induced skin infections in guinea pigs. A topical treatment with 2% of 6 in a vanishing cream base, administered 24 h postinfection applied five times daily for 4 days, significantly reduced the number and size of herpetic vesicles.


Mercaptoimidazolylpropionic acid hydrobromide. Inhibition of tadpole collagenase and related properties. A mercapto analogue of histidine (1), (RS)-2-mercapto-3-(5-imidazolyl)propionic acid (2), was prepared by treatment of (RS)-2-bromo-3-(5-imidazolyl)propionic acid with trithiocarbonate. Decomposition of the resulting intermediate with hydrochloric acid followed by Sephadex G-15 chromatography permitted isolation of 2 as a hydrobromide complex having unusual stability and properties as evidenced by IR and 1H NMR data. The potency of this complex in inhibiting tissue (Rana catesbiana) collagenase was estimated by radial diffusion assay. The amount of 2 required to produce 50% inhibition was 3.8 +/- 1.5 mM compared to 8.7 +/- 2.5 mM for cysteine. Preliminary tests of oxygen susceptibility, mutagenicity, and toxicity suggest that this substance may warrant study as a therapeutic agent for control of collagenase-linked corneal ulcerations.


Nucleoside 5'-monophosphate analogues. Synthesis of 5'-sulfamino-5'-deoxynucleosides. The synthesis of two new nucleotide analogues is described. 5'-Sulfamino-5'-deoxyadenosine (1) was prepared by reaction of 5'-amino-5'-deoxyadenosine with (CH3)3N.203, and 5'-sulfamino-5'-deoxythymidine (2) was prepared from 5'-amino-5'-deoxythymidine by a similar reaction. The 5'-sulfamino nucleosides are shown to be quite stable to hydrolysis in acidic or basic aqueous solution Tests show that these compounds do not inhibit the growth of Escherichia coli or L1210 cells at concentrations less than 10(-4) M. At 10(-4) M compound 2 was found to give 70% inhibition of the replication of herpes simplex virus (type 1) with no effect on host cell growth (CV-1 monkey line).


Synthesis of branched-chain apiosylpyrimidines and their inhibition of lymphocyte proliferation. Branched-chain nucleosides containing beta-D-apio-L-furanose were synthesized by condensation of the bis(trimethylsiyl) derivative of uracil (11), thymine (12), 5-fromouracil (13), and 5-iodouracil (14) with the protected 1-O-acetylapiose in the presence of a Friedel-Crafts catalyst. D-Apio-L-furanosyluracil (11) and D-apio-L-furanosylthymine (12) show immunosuppressive activity for rat T-lymphocytes stimulated to grow by phytohemagglutinin but exhibit no inhibitory activity against herpes simplex virus. Compounds 11--14 did not inhibit herpes simplex virus replication, while low inhibition was obtained with the one nucleoside containing D-apio-D-furanose, D-apio-D-furanosyladenine. D-Apio-L-furanosyl-5-bromouracil (13), D-apio-L-furanosyl-5-iodouracil (14), bromouridine, and iodouridine suppressed growth of human lymphoblasts significantly more than the nonhalogenated apiosylphrimidines.


Protection against 7, 12-dimethylbenz[a]anthracene-induced rat mammary carcinoma by infection with mouse xenotropic type C virus. A single ip inoculation of female, outbred Sprague-Dawley rats with a viable mouse xenotropic type C virus significantly reduced the incidence and/or retarded the development of mammary carcinoma induced by 7, 12-dimethylbenz[a]anthracene administered orally 7 days after virus. Although infectious virus could not be isolated from organs of infected rats, high titers of circulating and tumor-associated antibodies were detected against the viral internal core protein p30, and a low-grade antibody response to intact virus or envelope glycoprotein was found. Moreover, a cell-mediated immune response, measured by lymphocyte transformation, was detected with the use of intact virus but not with p30 antigen. No immunity developed after a single inoculation of UV-inactivated virus. These data indicated that inoculation of adult individuals of heterologous species with viable xenotropic mouse type C virus resulted in the rapid disappearance of infectious virus from the recipient, followed by the development of both humoral and cellular immunity to virion constituents. These events led, by unknown mechanisms, to the effective retardation of chemical carcinogenesis when infection preceded carcinogen administration.


Effect of reovirus infection on pulmonary tumor response to urethan in strain A mice. The effect of reovirus type 3 infection on the pulmonary adenoma response to urethan in strain A mice was examined. Urethan carcinogenesis in this system was suppressed from 30 to 60% when mice were exposed to reovirus either 6 days before, on the same day as, or 14 days after urethan administration. These findings suggested that reovirus infection interfered with the progression of urethan-induced pulmonary adenoma rather than the induction of lung tumors by urethan. When mice received multiple exposures to reovirus, the lung tumor response was enhanced. These findings indicated that reovirus infection in particular and virus infection in general may play an important role in the carcinogenic response to environmental chemicals.


Virus-specific markers and virus-like particles in cell lines of tumors produced by CELO virus in Syrian golden hamsters. Cell lines were established from 2 primary hepatocellular carcinomas (HC's) and 3 sarcomas produced in Syrian golden hamsters inoculated as newborns with chicken embryo lethal orphan (CELO) virus. Cell lines from 2 sarcomas (COT, CMT) and 1 HC (CEHEP) produced CELO virus-specific T-antigen. The antigen was not detected in cells of the third sarcoma line (RCT) until they had undergone more than 34 passages in vitro. Although 5-10% of cells in the second HC line (CILT/2) contained T-antigen during early passages, it was not demonstrable after the fifth subculture. Nevertheless, cells of both HC lines possessed CELO virus tumor-specific transplantation antigen. All 5 cell lines also contained hamster type R particles, and both HC lines had type C and intracytoplasmic type A particles. The percentage of carcinoma cells producing type R particles increased during cultivation in vitro, whereas the number of cells with type A particles decreased. Treatment with dibutyryl cyclic AMP and theophylline enhanced the number of cells producing type C particles in 1 HC line and type R particles in 2 sarcoma lines.


Delay of tumor growth in hamsters treated with lynestrenol and effect of Staphylococcus aureus Cowan A. The action of lynestrenol, a synthetic progesterone-like substance, was studied in Syrian golden hamsters inoculated with cells transformed by herpes simplex virus type I. Daily ip injections of 1 mg lynesterol/kg delayed tumor growth. Lynestrenol alone only slightly increased the survival of the animals. This effect was more marked when the animals were also given injections of Staphylococcus aureus Cowan A. Survivors showed an increased resistance to challenge with infected cells. Hamsters previously given injections of killed transformed cells and then challenged with living cells showed a facilitation of tumor growth. This facilitation was inhibited by lynestrenol alone or combined with S. aureus Cowan A.


Epstein-Barr virus infections in Brazil. II. Hodgkin's disease. Sixty-seven cases of Hodgkin's disease (HD) occurring in São Paulo, Brazil, were studied. Males with HD predominated over females 2.3 to 1. Sixty-six percent of the cases occurred in patients under 30 years of age, 31.7% under 20 years of age, and only 7.5% after 50 years of age. Lymphocyte predominance and mixed cellularity histologic types were most common in patients less than 15 years old, and nodular sclerosis was most common in the 15- to 19-year-old group. Sera from all patients had antibody to the viral capsid antigen (VCA) of Epstein-Barr virus (EBV). The geometric mean titer (GMT) of VCA antibody with the use of Jijoye cells as antigen was 1:162, and 31.3% of patients had titers of 1:320 or more; in controls, the GMT was 1:67 and 3.8% had titers of 1:320 or more. Similar results were obtained when EB-3 cells were used as antigen. The highest titers occurred in males, in mixed cellularity and lymphocyte depletion forms, and in stage 2 of illness. EBV-specific IgM antibody and heterophile antibody levels were not elevated, but 20.5% of the HD patients had antibody to the early antigen of EBV present in their sera. Antibody levels for herpes simplex virus, cytomegalovirus, rubella, measles, parainfluenza viruses, and papovavirus were not significantly elevated over those in matched controls.


Chromosome changes (trisomies #15 and 17) associated with tumor progression in leukemias induced by radiation leukemia virus. An experimental system was developed that permitted nonrandom chromosome changes that occur in radiation leukemia virus (RadLV)-induced lymphomas to be followed during tumor progression. RadLV variant-induced preleukemia and leukemia cells originating from female inbred C57BL/6 mice were injected into male animals of the same strain. Since all donors were females and all recipients were males, the sex chromosome complements (XX and XY) were used to distinguish the preleukemia and leukemia cells from those of host origin. The G-banding analysis revealed that more than 50% of animals that were inoculated with preleukemia cells and that developed leukemia possessed tumor stem-lines of 41 chromosomes with a tristomy of chromosome #15. In animals inoculated with overt leukemia cells and in which tumor progression occurred, the G-banding an additional trisomy of chromosome #17. The cytogenic data strongly suggested that the trisomy of chromosome #15 was the first specific tumor-associated chromosome change that occurred in the process of conversion of RadLV-induced preleukemia cells to fully autonomous tumor cells.


Quantitation of secretory component levels in cyst fluids, ascitic fluids, and sera from ovarian adenocarcinoma patients. A secretory component (SC) was detected by radioimmunoassay in the cyst fluids, ascitic fluids, and sera from patients with ovarian adenocarcinomas. Serous cyst fluids and ascitic fluids showed lower levels (expressed as means +/- SE) of SC (1.37 +/- 0.37 and 1.24 +/- 0.24 microgram/ml, respectively) than mucinous cyst fluids (181.50 +/- 50.40 microgram/ml). SC levels in the sera of all patients with ovarian adenocarcinoma were high (12.67 +/- 1.43 microgram/ml) when compared to SC levels in the sera of normal individuals (2.34 +/- 0.41 microgram/ml). Sera from patients with ovarian cancers diagnosed as serous, mucinous, papillary, and poorly differentiated adenocarcinomas showed SC levels of 9.93 +/- 1.68, 22.44 +/- 3.24, 7.35 +/- 1.13, and 10.10 +/- 1.58 microgram/ml, respectively.


Type B and type C RNA tumor virus replication in single cells. Electron microscopy with tannic acid. Simultaneous replication of murine mammary tumor virus (type B) and murine leukemia virus (type C) was demonstrated electron microscopically along continuous stretches of the plasma membrane of single cells in cultures ofthe Mm5mt/c1 cell line. Types B and C virus buds were discriminated in thin sections with the aid of a tannic acid fixative that revealed the type B surface spikes as a homogeneous band of intermediate density and constant width on the surfaces of some buds (type B), whereas others (type C) remained with relatively smooth envelopes. Both types B and C buds may contain morphologically identical horseshoe-shaped nucleoids. Therefore, their identify (type B or C) could be ascertained in thin sections only on the basis of recognition of surface spikes.


Pharmacology of the brachium conjunctivum: red nucleus synaptic system in the baboon. Acetylcholine, biogenic amines, and certain amino acids were applied by microiontophoresis to parvicellular and magnocellular red nucleus (RN) neurons of baboon while recording brachium conjunctivum (BC)-evoked and amino acid-evoked unit discharge from these neurons. Glycine, gamma-aminobutyric acid, and beta-alanine were potent depressants of BC-RN synaptic transmission, amino acid-evoked firing, and spontaneous activity of all RN neurons studied. Glycine was clearly more potent than the other 2 depressant amino acids. L-Glutamic and DL-homocysteic acid were strong excitants of all RN neurons tested. Dopamine, noradrenaline, and 5-hydroxytryptamine depressed the excitability of both parvicellular and magnocellular RN neurons; no excitatory effects were observed with these biogenic amines on RN neurons. Acetylcholine increased the rate of firing of spontaneously discharging parvicellular RN neurons and facilitated the amino acid-induced firing of these same neurons. Acetylcholine did not facilitate BC-RN synaptic transmission nor could this transmission be blocked by cholinergic antagonists. Unlike parvicellular RN neurons, the responsiveness of magnocellular neurons was either unaltered by acetylcholine or slightly decreased. These experiments demonstrate a difference in the pharmacologic responsiveness of parvicellular and magnocellular RN neurons to acetylcholine but do not provide evidence for a cholinergic input to RN via the brachium conjunctivum.


Aanlysis of dCMP deaminase and CDP reductase levels in hamster cells infected by herpes simplex virus. Several enzymatic activities involved in the biosynthetic pathways of nucleotides, including thymidine kinase, which has been used as a biochemical marker in studies of gene transfer, are induced by herpes simplex virus (HSV). The utility of additional markers prompted us to reanalyze the effects of HSV infection on the activities of two other enzymes for which direct selective methods can be devised: dCMP deaminase and CDP reductase. For this purpose, mutant Chinese hamster (lA1) cells devoid of dCMP deaminase activity or Syrian hamster (BHK-21/C13) cells were infected by HSV type 1 or 2, and the activities of thymidine kinase, dCMP deaminase, and CDP reductase were measured in the cell extracts. The reported induction of thymidine kinase and CDP reductase by HSV was confirmed, whereas the stimulation of dCMP deaminase activity could not be observed. For both cell lines, the HSV-induced CDP reductase differed from the host enzyme by sensitivity to inhibition by both dTTP and dATP. This property should be helpful in developing a selection system for this activity.


5'-terminal nucleotide sequences of mammalian type C helper viruses are conserved in the genomes of replication-defective mammalian transforming viruses. The RNAs of replication-defective murine and primate type C transforming viruses were analyzed for the presence of nucleotide sequences homologous to the genomes of their respective helper type C viruses by using DNAs complementary (cDNA) to either the 5'-terminal (cDNA5') or total (cDNAtotal) nucleotide sequences of the helper virus RNA. The defective viruses examined have previously been shown to vary in their ability to express helper viral gag gene proteins. With cDNAtotal as a probe, these transforming viruses were shown to vary in their representation of helper sequences (15 to 60% hybridization of cDNAtotal). In striking contrast, 5'-terminal-specific sequences of the helper virus were conserved in the RNAs of every transforming virus tested (is greater than 80% hybridization of cDNA5'). These findings suggest a critical role for these sequences in the life cycle of the defective transforming virus.


Detection of proviral DNA in horse cells infected with equine infectious anemia virus. Equine infectious anemia virus (EIAV) recently has been shown to possess a high-molecular-weight RNA genome and a virion reverse transcriptase. We completed the demonstration that EIAV is a retrovirus by showing the presence of proviral DNA in equine cells infected in vitro, but not in normal horse DNA. These studies were performed by using a highly representative cDNA probe synthesized by the virion polymerase. It was found that this cDNA reassociated extensively, and with high thermal stability, with either viral RNA or DNA extracted from infected cells, but showed no detectable reassociation with DNA from uninfected horse cells. Similarly, sequences related to EIAV were neither found in the DNA of four other Equus species, nor in a variety of other mammals including sheep, cows, pigs- dogs, cats, and humans; nor did EIAV cDNA hybridize with a variety of other retrovirus RNAs. These experiments were performed under conditions of very low stringency to enable detection of distantly related sequences, with a sufficient ratio of DAN to cDNA to allow detection of less than one viral copy per haploid genome. We conclude that EIAV is not an endogenous virus of the horse or of the other species tested.


Role of the simian virus 40 gene A product in regulation of DNA synthesis in transformed cells. Cells transformed by tsA mutants of simian virus 40 (SV40) are temperature sensitive for the maintenance of the transformed phenotype. The kinetics of induction of DNA synthesis were determined for hamster cell transformants shifted to the permissive temperature after a 48-h serum arrest at the nonpermissive temperature. DNAsynthesis was initiated in the tsA transformants by 8 h after shiftdown was maximal by 12 h. The presence or absence of fetal bovine serum at the time of temperature shift had no effect on the kinetics of initiation of DNA synthesis. Analysis of TTP in tsA transformants revealed similar levels of incorporation of [3H]thymidine into TTP at both permissive and nonpermissive temperatures. Autoradiography revealed that by 12 h after a shift to the permissive temperature, approximately 50% of the cells exhibited labeled nuclei after a 60-min pulse with [3H]thymidine, indicating that a majority of the cells were actively synthesizing DNA. By 8 to 12 h after a shiftup of confluent tsA transformants to the nonpermissive temperature, the number of labeled nuclei was reduced to approximately 16%, regardless of serum concentration. These data indicate that the SV40 gene A product, either directly or indirectly, regulates cellular DNA synthesis in transformed cells.


Genome organization of RNA tumor viruses. I. In vitro synthesis of full-genome-length single-stranded and double-stranded viral DNA transcripts. Genome-length complementary DNA (cDNA) transcripts were synthesized in vitro by using purified virions of avian myeloblastosis virus. Moloney murine leukemia virus, and clone 124 mouse sarcoma virus. The size of the genomelenth cDNA transcripts was measured on either alkaline sucrose gradients or alkaline agarose gels. The longest cDNA transcripts synthesized by using avian myeloblastosis virus, Moloney murine leukemia virus, and clone 124 mouse sarcoma virus were 7, 9 and 6 kilobases (kb), respectively. The in vitro system used was capable of synthesizing double-stranded DNA, but the plus strands (same polarity as the viral RNA) were only 0.5 to 1.5 kb long. Lone Moloney murine leukemia virus cDNA transcripts were used as templates to synthesize the second plus strand. Essentially two strategies were employed as follows. (i) The 3' ends of the cDNA transcripts were extended by addition of 50 to 100 dAMP residues by terminal deoxynucleotidyl transferase. The (dA)n-tailed cDNA transcripts were used as templates along with an oligomer of dT as primer and Escherichia coli DNA polymerase to synthesize the plus strands. (ii) DNase-digested calf thymus DNA was used to prime the synthesis of plus strands on long cDNA with E. coli DNA polymerase I. In both cases, the synthesis of the plus strands was monitored by increased resistance of the cDNA templates to single-strand-specific S1 nuclease. The double-stranded DNA was fractionated on neutral sucrose gradients. Analysis of the double-stranded DNA synthesized by using oligo(dT) primer showed the plus strands to be about 5 to 6 kb long, whereas the plus strands synthesized by using DNase-digested calf thymus DNA primers were only 0.3 to 0.5 kb long. Double-stranded DNA synthesized by either method has an average size of 6 x 10(6) daltons. Double-stranded DNA was also synthesized by using cDNA transcripts as templates without the addition of any primers. In this case, the plus strands were covalently linked to the template strand and were not representative of the whole parent strand.


Simian virus 40 DNA extracted from infected cells with sodium deoxycholate no longer reflects its in vivo superhelix density. When simian virus 40 DNA is extracted from infected cells with low concentrations of sodium deoxycholate, which selectively extract non-encapsidated simian virus 40 DNA, the DNA has a lower average number of superhelical turns than the DNA extracted from purified viral particles. During extraction, a partial deproteinization of the DNA by a concentration of detergent that did not inactivate a nicking-closing activity led to the removal of some superhelical turns. The DNA extracted in this way no longer reflected its in vivo number of superhelical turns.


Sequence of protein synthesis in cells infected by human cytomegalovirus: early and late virus-induced polypeptides. At least 10 distinct early virus-induced polypeptides were synthesized within 0 to 6 h after infection of permissive cells with cytomegalovirus. These virus-induced polypeptides were synthesized before and independently of viral DNA replication. A majority of these early virus-induced polypeptides were also synthesized in nonpermissive cells, which do not permit viral DNA replication. The virus-induced polypeptides synthesized before viral DNA replication were hypothesized to be nonstructural proteins coded for by the cytomegalovirus genome. Their synthesis was found to be a sequential process, since three proteins preceded the synthesis of the others. Synthesis of all early cytomegalovirus-induced proteins was a transient process; the proteins reached their highest molar ratios before the onset of viral DNA replication. Late viral proteins were synthesized at the time of the onset of viral DNA replication, which was approximately 15 h after infection. Their synthesis was continuous and increased in molar ratios with the accumulation of newly synthesized viral DNA in the cells. The presence of the amino acid analog canavanine or azetadine during the early stage of infection suppressed viral DNA replication. The amount of viral DNA synthesis was directly correlated to the relative amount of late viral protein synthesis. Because synthesis of late viral proteins depended upon viral DNA replication, the proteins were not detected in permissive cells treated with an inhibitor of viral DNA synthesis or in nonpermissive cells that are restrictive for cytomegalovirus DNA replication.


Genetic analysis of a cytomegalovirus-like agent isolated from human brain. An unusual cytomegalovirus (CMV, strain Colburn) isolated from brain biopsy of a boy with clinical encephalopathy was studied for genetic relatedness to human and simian CMV. Cross-examination of the purified viral DNA by DNA-DNA reassociation kinetics analyses showed more than 90% homology between Colburn virus and simian CMV (strain GR2757) and a lack of detectable homology between Colburn virus and human CMV (strains AD-169 and TW-87). Restriction endonuclease analysis of Colburn DNA showed some similarity of the DNA fragment pattern with that of simian CMV DNA, although the DNA fragment patterns were not identical, and showed no similarity to that of human CMV DNA. The molecular size and density of viral DNA were close to those of simian CMV DNA. The antigenic study, as performed by complement fixation and neutralization tests, showed strong cross-reactivity of Colburn virus to simian GR2757 virus. One-way cross-reaction of Colburn virus to several human CMV isolates (AD-169, Davis, and Town) was detected by complement fixation; this one-way cross-reaction was not obvious in a plaque neutralization test. It was concluded that Colburn is a simian CMV-related virus.


Isolation of the envelope of vesicular stomatitis virus. Vesicular stomatitis virus was disrupted by a combination of freezing and thawing, osmotic shock, and sonic treatment. Subviral components were separated by isopycnic centrifugation. The low-density, lipid-rich fractions were pooled and shown to contain primarily viral glycoprotein. Further purification of this material resulted in the isolation of a preparation of vesicles which contained only the G protein and the same phospholipids as in the intact virions and exhibited spikelike structures similar to those on intact vesicular stomatitis virions. We conclude that we have isolated fragments of native vesicular stomatitis virus envelopes.


Peptide analysis of the transformation-specific antigen from avian sarcoma virus-transformed cells. Sera from rabbits bearing tumors induced by avian sarcoma virus (ASV) were ussed to immunopecipitate virus-specific proteins from extracts of chicken, hamster, and field vole cells transformed by ASV. Two virus-specific proteins having molecular weights of 76,000 and 60,000 were found in all cell lines examined. The 76,000-molecular-weight protein, Pr76, is the precursor to the internal core proteins of ASV. The 60,000-molecular-weight (60K) transformation-specific antigen from each cell line was subjected to peptide analysis, using chymotrypsin and Staphylococcus aureus V8 protease. The resulting peptide maps of the 60K protein from the different ASV-infected cell types were similar for each enzyme, strongly suggesting that the 60K protein is virus coded. Two-dimensional analysis of chymotryptic peptides from Pr76 and 60K reveals that 60K is not related to the gs antigen precursor. Radiolabeling of ASV-transformed cells with inorganic phosphate revealed that 60K is phosphorylated in vivo. The 60K proteins isolated from both ASV-transformed chicken and field vole cells were found to contain one tryptic phosphopeptide. The tryptic phosphopeptides of 60K from both cell lines migrated identically upon two-dimensional peptide analyses, and their migration differed from that of the principal phosphopeptide of Pr76.


Human cytomegalovirus DNA. I. Molecular weight and infectivity. Human cytomegalovirus DNA (strain AD 169) was isolated from purified virions and further purified by sucrose density gradient centrifugation. The viral DNA molecules were studied by electron microscopy and found to be linear and to have a length of 76.22 +/- 5.22 micron, corresponding to a molecular weight of 147 +/- 6.2 x 10(6). The DNA was infectious when tested in human embryonic lung cells.


Anomalous behavior of the major avian myeloblastosis virus glycoprotein in the presence of sodium dodecyl sulfate. The sodium dodecyl sulfate (SDS) complex of the major glycoprotein of avian myeloblastosis virus exhibited an anomalously low free electrophoretic mobility compared with those of non-glycosylated protein standards. The apparent molecular weight of the glycoprotein calculated from the relation between log molecular weight and electrophoretic mobility depended on the acrylamide concentration and reached a lower limit of 80,000. The molecular weight was also estimated from the retardation coefficients of protein standards and the viral glycoprotein. This method yielded a molecular weight of 64,000 for the avian myeloblastosis virus glycoprotein. When gel chromatography in SDS was used to determine the apparent molecular weight of the glycoprotein from its hydrodynamic properties alone, the estimated value was 50,000. The generally assigned value of 80,000 daltons for the avian myeloblastosis virus major glycoprotein, as determined by SDS electrophoresis, may be an overestimate due to its relatively low free electrophoretic mobility and peculiar conformation in SDS.


Renal tumors involving the inferior vena cava: plan for management. Our experience in the management of 12 malignant renal tumors involving the inferior vena cava is analyzed and a plan to manage the different degrees of caval involvement is discussed. The use of Pringle maneuver to control hepatic circulation during removal of suprahepatic caval extensions is recommended. Followup to date supports aggressive operative management in these patients.


Surgical considerations in treatment of intraductal carcinoma of the prostate. Intraductal carcinomas of the prostate comprise about 3 per cent of all prostatic carcinomas and constitute a heterogeneous group of tumors that include 1) transitional or squamous cell carcinoma, 2) intraductal adenocarcinoma, 3) endometrioid carcinoma and 4) mixed ductal carcinoma. Generally, these tumors are poorly responsive to endocrine and radiation therapy, and complete surgical excision offers the best chance for long-term survival. A case of intraductal adenocarcinoma is presented that illustrates many of the features of these tumors.


Gallium scans for staging small cell lung cancer. The high incidence of gallium 67 accumulation in lung cencer has made radioistope scanning with this agent useful in identifying the extent of cancer locally. However, we investigated the usefulness of whole-body gallium 67 scanning, compared with physical examination, bone, liver and brain scans, and bone marrow aspirate and biopsy, in detecting metastases outside the chest in 47 patients with small cell lung cancer. In each case whole-body scanning with gallium 67 was inferior to the other methods used to detect extrathoracic tumor deposits.


Establishment of cultured cell lines derived from a human gastric carcinoma. A human gastric carcinoma cell line, KATO-II, and its subline, KATO-III, have been established in vitro from a pleural effusion of a 55-year-old male patient. Kato-II cells have been maintained using a culture medium containing human cord serum and KATO-III cells have been cultured with a medium containing fetal bovine serum. Both cell lines are morphologically quite similar; they grow in vitro floating free and have cytological features of signet ring cells. Population doubling times for KATO-II and KATO-III were 74 hours and 36 hours, repsectively. The modal chromosomal numbers of these cell lines fell in a tetraploid range. Heterotransplantation of KATO-II and KATO-III cells was successfully done in the cheek pouches of antithymocyte serum-treated hamsters. Microscopical appearance of the resultant tumors was consistent with poorly-differentiated adenocarcinoma comparable to the original tumor.


[Aspects of enzymatic toxicology of metals. A review (author's transl)]. The adverse and beneficial effects of metals have occupied a great concern in many branches of biology for centuries, but their biochemical roles have been studied systematically only in the last four decades. Almost all the metals, especially those of high toxicity, bind readily to mercapto groups, and these are frequently important in enzyme systems. Furthermore, a large number of in vitro and in vivo studies have shown that toxic metals can replace essential metals in many of their metalloenzymes, with resultant changes in activity. Metals also bind to protein, phospholipid and nucleic acid, and have been shown to effect a change in the conformation of enzymes required for normal function, or to uncouple oxidative phosphorylation. This review, of necessity brief, will focus on the biochemical and enzymatic effects of metals, emphasizing meanings available to occupational medicine. The present knowledge on enzymatic toxicology of metals can provide for access to three main problems. These include an exposure evaluation, a health injury evaluation and a pathogenic understanding of workers exposed to metals. In this article, these problems are discussed in general, and the recent developments made in the enzymatic toxicology of cadmium and lead are presented with pertinent literatures.


Familial hypercholesterolemia: pathogenesis of a receptor disease. Familial hypercholesterolemia is a prototype for a class of diseases that result from defects in receptor molecules. The three cardinal features of familial hypercholesterolemia are: 1) a selective elevation in the plasma level of one cholesterol-carrying lipoprotein, low density lipoprotein (LDL); 2) a selective deposition of LDL-derived cholesterol in macrophage-like scavenger cells throughout the body, but not in parenchymal cells; and 3) inheritance as an autosomal dominant trait with gene dosage effect, i.e., the disease is more serious in patients with the homozygous than with the heterozygous state. In this article, we review the evidence that each of these cardinal features of familial hypercholesterolemia can be explained by a genetic defect in a cell surface receptor for plasma LDL.


[New approaches to the evaluation of the cholesterol content of plasma lipoprotein fractions in normo- and hyperlipoproteinemias]. The examination of 1073 males ranging in age from 40 to 59 living in Moscow and Leningrad, and of 146 patients with ischemic heart disease showed a decrease in the alpha-lipoprotein cholesterol level and an increase in the cholesterol content of atherogenic fractions (beta- and pre-beta-lipoproteins) in individuals with ischemic heart disease in the absence of hyperlipoproteinemia (HLP). In combination of ischemic heart disease with hyperlipoproteinemia these changes are pronounced to a larger degree. From the analysis of the results of the studies it is suggested that the coefficient (see article) is determined to reveal disorders in lipoprotein metabolism particularly in the absence of hyperlipoproteinemia. This coefficient is significantly higher in individuals with ischemic heart disease and no hyperlipoproteinemia than in healthy males of the same age group; its value is highest in ischemic disease with hyperlipoproteinemia.


[The disease pattern of the diffuse, nesidioblastic hyperplasia of the pancreatic islets in newborn and infants (author's transl)]. The course of illness of a male infant who lived for seven months with a diffuse, nesidioblastic hyperplasia of pancreatic islets is described. Before surgical intervention the diagnosis should be ascertained by 1. observation of acetonuria which is always absent after hypoglycemic episodes, 2, the typical constellation of insulin concentration, free fatty acid concentration and beta-hydroxybutyrat during a hypoglycemia (as found by Baker et al. 1976) and/or by simultaneous measuring of glucose-insulin levels under the conditions of fasting as well as of oral leucine, oral glucose and intravenous tolbutamide loading. Therapy with diazoxide should be tried in any case. If all conservative measure fail and relative or absolute hyperinsulinemia is proved, experience shows that an immediate operation is indicated.


[New aspects of catecholamin-receptor interactions. Pathophysiological and clinical implications (author's transl)]. The molecular aspects of catecholamine-receptor interactions are reviewed with respect to their clinical implications. Beta- and in some tissues alpha-adrenergic receptors are coupled to the membrane-bound adenylate cyclase. The adrenergic receptor sites are distinct membrane constituents. Their number and the ratio alpha-to beta-receptor site depend on the plasma concentration of catecholamines and are affected by diet and endocrinological factors. Recent clinical studies suggest that long-term treatment with beta-adrenergic agonists may induces desensitization of target tissues due to changes in the adrenergic receptor moieties.


Responsiveness of plasma aldosterone: dependency upon basal secretory activity. The responsiveness of plasma aldosterone levels to various stimuli was evaluated in 44 normal subjects, 17 patients with mild to moderate renal failure, 30 patients with terminal renal failure, and 13 anephric subjects. Plasma aldosterone, renin activity (PRA), cortisol, sodium, and potassium levels were measured before and after one hour of upright posture (N = 191); ACTH infusions (N = 76); and angiotensin II infusion (N = 36). Plasma aldosterone responses correlated (r greater than or equal to 0.53; p less than 0.02) with basal plasma aldosterone levels during upright posture in all four groups, with ACTH infusion in all groups except anephric subjects, and with angiotensin II administration in patients with mild to moderate renal failure or patients combined. These relationships were consistently closer than those between aldosterone responses and changes in PRA or basal PRA. However, postural aldosterone responsiveness at any given basal aldosterone level was significantly lower in patients with renal disease than in normal subjects, and this was associated with a parallel impairment in renin responsiveness. In contrast, when related to basal levels aldosterone responsiveness to ACTH or angiotensin II appreared to be comparable in normal subjects and patients with renal disease. Aldosterone responses to posture, ACTH, or angiotensin II did not correlate with associated changes in plasma cortisol, sodium, or potassium levels. These data suggest that basal adrenal secretory activity is a major factor conditioning aldosterone responsiveness to various stimuli in normal subjects as well as in patients with renal disease.


Oral hydration rotavirus diarrhoea: a double blind comparison of sucrose with glucose electrolyte solution. Of 57 male children, aged 5 months to 2 1/2 years with rotavirus diarrhoea, 28 were given oral therapy with sucrose electrolyte solution and 29 were given glucose electrolyte solution in a randomised double-blind trial. All were rehydrated and remained so on oral therapy alone. These patients were compared with 44 children, also with rotavirus, who were treated only with intravenous hydration. The oral therapy and intravenous therapy groups did not differ clinically in the rate of rehydration or the rate of purging. Vomiting did not prevent the giving of oral therapy during hospital admission. Bangladeshi children with rotavirus diarrhoea have a defect of carbohydrate digestion but this defect does not prevent the use of a sugar electrolyte solution for oral hydration.


Nasopharyngeal angiofibroma: a twenty year study. A control rate of approximately 80% has been obtained in 45 cases of juvenile nasopharyngeal angiofibroma treated by a single moderate dose of radiotherapy at the Princess Margaret Hospital, Toronto. The remaining patients ultimately achieved control generally after further irradiation. Immediate side-effects of the treatment have been minimal, and no late complications have been observed. In 2-20 years' follow-up, no irradiation-induced tumors have been found. Superiority of irradiation in these cases over surgery is emphasized, as well as the importance of using sophisticated techniques to limit the volume of tissues irradiated and protect vulnerable radiosenstitive structures. Because tumor regression can be expected to continue for many months after treatment has been completed, a policy of observation following radiotherapy is recommended.


Recurrent laryngeal nerve pathology in spasmodic dysphonia. Since it was first described in 1871, spasmodic (spastic) dysphonia has been considered a disease of psychogenic origin. Unsupported theories of possible organic etiology have appeared sporadically in the literature. In 1976 sectioning of the recurrent laryngeal nerve for patients with this disease was reported with resultant improvement in voice production. This was attempted because the spasmodic dysphonic has, in effect, already compensated vocal cords bilaterally. It was reasoned, therefore, that if one of these was paralyzed the patient would immediately be converted to a state approximating that of well-compensated unilateral vocal cord paralysis which situation, as is well known, usually carries with it a fairly good voice. A controlled study to evaluate the efficacy of this surgical approach has been undertaken at the Cleveland Clinic during the past year. In an attempt to elucidate the possible organic etiology of spasmodic dysphonia, a section of nerve was removed in every case and examined by both light and electron microscopy. Special stains for myelin were also used on the light microscopy specimens. Demyelinization has been found in most of the cases examined by electron microscopy. Possible correlation between this disease entity and other cranial nerve syndromes of unknown etiology is noted. Such conditions as trigeminal neuralgia, glossopharyngeal neuralgia, belpharospasm, hemifacial spasm, and even possibly Bell's palsy may exhibit a similar etiology.