Cocaine and succinylcholine sensitivity: a new caution. Cocaine was stable (93.6 +/- 5.0 (SD)% remaining after 120 minutes) when incubated in plasma from six patients with histories of succinylcholine sensitivity and low dibucaine numbers (phenotype A), but was rapidly hydrolyzed (40.6 +/- 6.6% remaining after 120 minutes) when incubated in plasma obtained from normal subjects (phenotype U). Intermediate decay (69.4 +/- 9.0% remaining) occurred in three heterozygous offspring of the succinylcholine-sensitive patients. Since serum cholinesterase has recently been implicated in the human metabolism of cocaine, patients who are homozygous for the atypical enzyme may be at greater risk for a toxic reaction when administered this drug. We recommend caution in using cocaine for topical anesthesia in patients with histories of succinylcholine sensitivity or low dibucaine numbers.


Platelet aggregation during and after general anesthesia and surgery. Because of evidence suggesting that volatile anesthetics interfere with platelet aggregation in vitro, platelet function was investigated in patients exposed to combinations of nitrous oxide, halothane, and enflurane during brief surgical procedures. In 12 patients undergoing elective operations, blood was sampled before anesthesia, after induction but prior to surgery, during surgery, and postoperatively. Platelet aggregation by ADP, epinephrine, and collagen was similar during each sampling period in all but two subjects in whom only minor changes were noted. During brief exposure to these general anesthetics, there appears to be no clinically important inhibition of platelet function.


Survey of infants born in 1973 or 1975 to Swedish women working in operating rooms during their pregnancies. The outcome of deliveries during 1973 and 1975 of women working during their pregnancy in operating rooms, using register data that provided information on both occupation and outcome of delivery was studied. There were 494 women who worked throughout pregnancy, 37 women worked for more than half of their pregnancies, and 10 women worked during less than half of their pregnancies. The study groups were compared with a reference population composed of all women employed in medical work in Sweden who were delivered during the 2 years. The study population differed from the reference population with respect to maternal age (and perhaps also to parity); corrections were made for this. No differences in the incidence of threatened abortions, in birth weight, in perinatal death rate, or in the incidence of congenital malformations were found. Possibly more gestations lasting less than 37 weeks were noted. We suggest that the negative outcome of the study is due to the unbiased data collection.


A parascalene technique of brachial plexus anesthesia. A technique for brachial plexus anesthesia is described consisting of the injection of 20 ml of local anesthetic solution into the lower part of the posterior triangle of the neck at a point 1.5 to 2 cm above the clavicle at the lateral border of the anterior scalene muscle. The technique is simple, safe, and produced satisfactory anesthesia of the entire extremity in 97 of the first 100 cases in which it was used. Side effects and complications were minor and transient.


Effects of diazepam on succinylcholine-induced myalgia, potassium increase, creatine phosphokinase elevation, and relaxation. Diazepam in a dose of 0.05 mg/kg was studied to determine its effect on a subsequently administered dose of 1 mg/kg of succinylcholine. This dose of diazepam prior to succinylcholine (1) significantly diminished the incidence of postoperative muscle pain; (2) decreased the usual increase in serum potassium; (3) did not prevent the rise in creatine phosphokinase; (4) reduced the incidence of muscle fasciculation; and (5) did not affect the magnitude or duration of the succinylcholine neuromuscular block. It was concluded that diazepam had several advantages over d-tubocurarine in the prevention of succinylcholine-induced muscle pain.


Local anesthetics and pulmonary venous admixture in pregnant and nonpregnant sheep. Pulmonary venous admixture (Qs/Qt) was determined in nine nonpregnant sheep and in seven pregnant sheep during their last trimesters of gestation. Pulmonary venous admixture was greater in the pregnant animals (5.7 +/- 1.6% of the cardiac output; mean +/- SD) than in the nonpregnant ones (4.2 +/- 1.3%; p less than 0.05). On consecutive days, four local anesthetics were infused in random order via a femoral venous catheter into each sheep for 30 minutes. Total doses of 1.8 mg/kg of bupivacaine, 2.7 mg/kg of etidocaine, 6 mg/kg of lidocaine, and 15 mg/kg or chloroprocaine were administered. None of the anesthetics induced a significant change in pulmonary venous admixture.


Physiologic effects of deep hypothermia and microwave rewarming: possible application for neonatal cardiac surgery. Deep hypothermia (20 C) without cardiopulmonary bypass is a valuable technique during cardiac surgery in infants but rewarming of the heart following circulatory arrest and cardiac repair has traditionally been a lengthy and difficult process. In experimental animals rewarming the heart with microwave energy, as reported in this work, warms the heart before warming the periphery. In 18 mongrel dogs that were surface cooled to 20 C, we found that during microwave rewarming the core temperature rose 4.7 C per hour. Whole body oxygen consumption, heart rate, and cardiac output returned to normal at rates equal to the rates at which they decreased during surface cooling. Blood pressure and arterial gases remained adequate. Microwave rewarming appears to be a useful method for reestablishment of cardiac function and normothermia following deep hypothermia.


Recovery from general anesthesia: a simple and comprehensive test for assessment. A procedure of totaling the value of coins of different denominations has been evaluated as a simple test for assessing recovery from general anesthesia. The procedure required the patient to pick up speedily seven coins of small denominations one after the other, name the value of each coin as it was picked up, add its value to that of the previous ones and obtain the cumulative total of the seven coins. The test simultaneously assesses return of sensory perception, acuity of vision, ability to quickly change eye fixation, fine muscle coordination, higher intellectual faculties required for mathematical performance, sustained concentration, and word naming. Its preliminary application in a small number of patients has shown consistent results.


Do volatile anesthetics act as ideal gases? Under conditions of temperature and pressure commonly found in operating rooms, the anesthetics enflurane, halothane, and isoflurane act as nearly ideal gasses even at concentrations producing a saturated vapor phase. The deviations from ideality are 3.44 +/- 0.44% (SEM) for enflurane, 0.94 +/- 0.30% for halothane, and 2.96 +/- 0.29% for isoflurane. That is, 0.9 to 3.4% more anesthetic is vaporized than would be predicted from the vapor pressure of these agents.


Distribution of serotypes of the Mycobacterium avium-intracellulare-scrofulaceum complex in Georgia. Cultures of the Mycobacterium avium-intracellulare-scrofulaceum (MAIS) complex, isolated from patients who lived throughout the state of Georgia, were studied using the serotyping scheme of Schaefer. One hundred four (77 per cent) of the 135 isolates tested could be classified into 16 serotypes. The rest were not typable. Fifty-eight per cent of all the typable strains were serotypes 1, 14, 16, and 42. Results of the serotyping were recorded, mapped, and analyzed by county of residence. Most frequent isolations of these mycobacteria were from patients who resided in the coastal plain of Georgia. This finding correlates with the mycobacterial skin test studies of Edwards and associates, who found that the largest percentage of reactors to PPD-Battey resided in the costal plain. The disparate distributions of certain serotypes suggest the possible importance of regional factors that may affect the serotypes of mycobacteria indigenous to an area. The data do not suggest that chicken or swine reservoirs are highly significant sources of human mycobacteria.


Trochlear nerve palsy in a case of herpes zoster ophthalmicus. A case of trochlear nerve palsy in herpes zoster ophthalmicus is presented. The rarity of fourth nerve involvement in herpes zoster is emphasized.


A clinical trial with Piloplex--a new long-acting pilocarpine compound: preliminary report. A clinical comparison of the effect of Piloplex (an emulsion containing a new pilocarpine polymer compound) to that of pilocarpine hydrochloride 2% q.i.d.hloride on the intraocular pressure (IOP) was performed on 93 eyes of 51 open-angle glaucoma cases. Clinical studies showed that Piloplex produced a prolonged therapeutic effect, this being consistent with the slow release pattern it has displayed during in vitro experiments. Comparison of diurnal IOP measurements of 84 eyes have shown that when treated with Piloplex, 11 b.i.d., 58 eyes (69%) were under control (IOP values less than 20 mm Hg), whereas only 34 of these 84 eyes (40.5%) were under control when treated with pilocarpine hydrochloride 2% q.i.d. The overall mean diurnal IOP value of the 84 eyes during the treatment with Piloplex, 11 b.i.d., was lower by 2.89 mm Hg (P less than 0.005) than the overall mean diurnal IOP of the same eyes while treated with pilocarpine hydrochloride 2% q.i.d.


[Comparison of the action of carminomycin and rubomycin on the dynamics of the primary and secondary immune responses]. The effect of rubomycin and carminomycin on the dynamics of the primary and secondary immune response and formation of the immunologic memory to sheep red cells in mice was studied. Differences in the character of the antibiotics effect indicative of the higher selective action of carminomycin on multiplying cells, precursors of the antibody-forming plasmids, were found. Theoretically interesting discrepancies in the effect of the antibiotics on the content of the antibodies in the serum and the antibody-producing cells in the spleen were shown. It was demonstrated that carminomycin had no effect on formation of the immunologic memory inspite of a noticeable decrease in the total number of the spleen nuclear cells and the number of the antibody-forming cells at the moment of immunization under the effect of the antibiotic.


Bacteriological survey of sixty health foods. A bacteriological survey was performed on 1,960 food samples encompassing 60 types of health foods available in the Baltimore-Washington, D.C., metropolitan area. No consistent bacteriological distinction (aerobic plate counts, total coliform and fecal coliform most probable numbers) was observed between foods labeled as organic (raised on soil with compost or nonchemical fertilizer and without application of pesticides, fungicides, and herbicides) and their counterpart food types bearing no such label. Types and numbers of samples containing Salmonella were: sunflower seeds, 4; soy flour, 3; soy protein powder, 2; soy milk powder, 1; dried active yeast, 1; brewers' years, 1; rye flour, 1; brown rice, 1; and alfalfa seeds,1. The occurrence of this pathogen in three types of soybean products should warrant further investigation of soybean derivatives as potentially significant sources of Salmonella.


Effect of inorganic sulfide on the growth and metabolism of Methanosarcina barkeri strain DM. Minimal growth of Methanosarcina barkeri strain DM occurred when sulfide was omitted fromthe growth medium, and addition of either sodium sulfate or coenzyme M to sulfide-depleted media failed to restore growth. Optimal growth occurred in the presence of 1.25 mM added sulfide, giving a molar growth yield (YCH4) of 4.4 mg (dry weight) of cells per mmol of CH4 produced. Increasing sulfide to 12.5 mM led to decrease in YCH4 (1.9 mg [dry weight]/mmol of CH4), in the specific growth rate and in be intracellular levels of adenosine triphosphate. However, the specific rate of methane production increased. The data suggested that at elevated sulfide levels (12.5 mM) the decrease in YCH4 might be a result of an increase in the relative energy needed for maintnenace and of uncoupling of growth from energy production.


Topical heat therapy for cutaneous chromomycosis. An aged man with a large scaly plaque over the left buttock was successfully treated by local heat therapy using an electric bed-warmer with a surface temperature of 46 degrees C. Clinically, the lesion involuted completely after two months of treatment. This was concomitantly confirmed by histopathological and mycological studies of biopsy specimens obtained from the lesion at an interval of several weeks. The isolated fungus strain of Fonsecaea pedrosol could grow at 37 degrees C on Sabouraud's dextrose agar, but was totally eradicated 24 hours after the culture was incubated at 46 degrees C. We consider that local heat therapy carried out under strict supervision is a quite effective treatment for chromomycosis. Moreover, it can be performed safely and without much discomfort to patients, and can yield satisfactory results in a relatively short time.


Mourning by the family after a stillbirth or neonatal death. Failure of the parents to mourn after the sorrow of a stillbirth or neonatal death can result in dire consequences for the well-being of the family. Doctors and nurses should learn how to facilitate mourning and should accept the strange and sometimes bizarre forms this may take. When mourning is facilitated, the family is likely to adjust better to its bereavement.


Ventricular pseudoaneurysm after transaortic septal myotomy for hypertrophic subaortic stenosis. Clinical and morphological features are described in a man in whom a pseudoaneurysm arising from the ventricular septum developed after transaortic septotomy for hypertrophic subaortic stenosis. Pseudoaneurysm of the left ventricle or ventricular septum has not been described previously in a patient with hypertrophic cardiomyopathy in whom only the transaortic approach was utilized for septotomy or septectomy or both. In addition, no previous studies have reported pseudoaneurysm arising from the ventricular septum.


Hemodynamic effects of various modes of cardiac pacing after operation for idiopathic hypertrophic subaortic stenosis. The hemodynamic effects of varying heart rate and pacing site were studied in 6 patients with idiopathic hypertrophic subaortic stenosis following operative relief of outflow obstruction. Ventricular pacing (117 beats per minute) resulted in a 26% decrease in cardiac output (p less than 0.02), a 54% increase in pulmonary capillary wedge pressure (p less than 0.03), and a 23% decrease in mean blood pressure (p less than 0.05), compared with normal sinus rhythm (88 beats per minute). Slow atrial pacing (112 beats per minute) did not significantly alter any hemodynamic variable compared with normal sinus rhythm. Rapid atrial pacing (143 beats per minute) produced a similar degree of hemodynamic impairment as ventricular pacing. This study demonstrates that ventricular pacing at heart rates commonly used clinically and rapid atrial rates result in a significant fall in cardiac output. Preservation of atrial systole at heart rates that allow adequate diastolic ventricular filling of a hypertrophied, noncompliant ventricle is stressed. In addition, atrial electrodes are useful to record atrial electrograms or induce rapid atrial stimulation to treat supraventricular tachyarrhythmias.


Pulmonary valve replacement with a porcine aortic heterograft. Pulmonary valve replacement is an uncommon surgical procedure, yet lifesaving when performed under the appropriate circumstances. The patient reported on here had the classic indications for valve replacement: increased pulmonary vascular resistance and associated pulmonary valve incompetence following repair of a congenital heart defect resulting in right heart failure and secondary tricuspid insufficiency. A review of the literature provides the basis for the decision to replace the pulmonary valve with a glutaraldehyde-preserved porcine heterograft.


Prevention of air embolism after mitral valve replacement with a porcine heterograft prosthesis. The production of prosthetic valve incompetence during atriotomy closure is among the operative maneuvers utilized to prevent air embolism in mitral valve replacement. The leaflets of a porcine bioprosthesis may be retracted safely and effectively, thereby producing temporary valve incompetence, by placing three polypropylene sutures through the valve orifice and around the sewing ring to encircle the leaflets. These traction sutures are eaily pulled out through the atriotomy suture line after all air has been displaced from the heart. The technique has been effective, easily accomplished, and without complications.


Long-term sequential hemodynamic evaluation of right ventricular outflow tract reconstruction using a valve mechanism. From May, 1972, to May, 1978, right ventricular outflow tract reconstruction was performed in 20 patients with congenital heart disease. A monocusp patch constructed entirely of glutaraldehyde-stabilized calf pericardium, was employed in 19 patients, and a composite conduit consisting of a three-cusp pericardial xenograft valve in a Dacron tube was used in 1 patient. There were 2 hospital deaths (10%) and no late deaths. The follow-up ranged from 5 to 75 months (mean, 55.7 +/- 4.6 months; total, 1,002 patient months). Eleven patients had completed 5 years of follow-up at the time of writing. Hemodynamic studies were performed in 10 patients at an average of 40 months after operation and the right ventricle-pulmonary artery systolic gradient was 9.3 +/- 1.7 mm Hg. This was not significantly different in 6 patients who had a second postoperative catheterization at 51.8 +/- 2.0 months following operation. Angiography showed fully mobile, thin valve cusps. These results compare well with those reported with other types of conduits, particularly with reference to relief of obstruction.


Hepatic damage produced by long-term alcohol consumption in well-nourished rats. A study of the effect of long-term alcohol consumption on the liver of well-nourished rats is described. Rats fed for 16 weeks on a semipurified diet supplemented with high levels of vitamins and lipotropic factors and alcohol corresponding to 35% of the total caloric intake developed marked fatty changes of the liver. Mild fatty changes were observed in pair-fed controls receiving as isoenergetic equivalent of sucrose instead of alcohol. Intracellular hyaline bodies, corresponding ultrastructurally to giant mitochondria were abundantly found in the hepatocytes of alcoholic rats, while in the controls they were not seen. Te findings in this investigation are postulated to provide further evidence that the long-term intake of alcohol exerts a direct causative role in the pathogenesis of liver damage.


[Possibilities of ovulation synchronization in puberty-induced gilts]. Puberty was induced in 39 clinically prepuberal gilts (two groups of three sub-groups each) by parallel but locally separated application of 500 IU PMSG ("Maretropin") and 250 IU HCG ("Gonadex"), with the view to testing ways to synchronise ovulation. Seventy-two hours were allowed to elapse, before 24 animals received another application of 500 IU HCG and 15 animals 250 IU HCG. The animals were slaughtered in consecutive groups of study ovulation and histolotically examined to disclose endometrial processes. Ovulations were found to be well synchronised in the recipients of a second injection of 500 IU HCG. Only sub-threshold effects with no synchronised ovulation were recorded from the animals that had received a second dose of 250 IU HCG. A second injection of 500 IU HCG should be given not until something between 78 and 82 hours after puberty induction for optimum follicle maturation and adequate proliferation of the endometrium.


[Morphokinetic effect of PMSG and sex steroids on parts of the cerebral-hypophyseal-gonadal axis of anestrous sheep]. The action of gonadotropins and of sexual steroids on links of the cerebro-hypophyseo-gonadal axis can be morphologically quantified in sheep. In that context, karyovolumetry is a highly dependabel and information-yielding quantitative morphological method. In studies conducted with anoestrous sheep and described in this paper that method has helped to detect an inhibitory effect of PMSG and progesterone upon the activity of neurons of the medial preoptic nucleus of the hypothalamus. The two sexual steroids produced even stronger inhibition of neuronal activity of the arcuate nucleus of the hypothalamus. Such results, when evaluated and re-used under the adequate test conditions might be helpful in identifying structures of central nervous disorders along the cerebro-hypophyseo-gonadal axis and produce reliable information on acting points or general action (activation or inhibition) of active substances as well as on the course of control and regulatory mechanisms.


[Trials of rut induction with prostaglandin F2alpha in gilts]. The possibility of oestrus induction by means of F-2-alpha prostaglandin was checked on nine gilts. The animals received intramuscular injections of 7.5 mg of 10 mg of PG F2alpha on the tenth or twelfth day of cycle. The sexual behaviours of the animals was followed up, and after slaughter followed macroscopic assessment of the ovaries as well as histomorphological and histochemical examinations of the uterus wall. The results have shown that no oestrus or ovulation could be induced by such injection on the tenth or twelfth day of cycle. Regression of corpora lutea and follicle growth five days following application on the twelfth day of cycle were somewhat faster than the same phenomena when observed following injection on the tenth day. However, oestrus and ovulation failed throughout to occur earlier than they would have occurred any way without any treatment. The cycle was not shortened by application of F-2-alpha prostaglandin.


The coupling of transcription from influenza virions to translation in vitro. The optimum conditions for the coupling of fowl plague virus (FPV) transcription to an in vitro reticulocyte translation system have been established and shown to be close to those required for maximum RNA synthesis by purified FPV virions. Products have been characterized by the peptides they yield on limited proteolysis in SDS and it has been shown that virus nucleoprotein (NP) and matrix (M) protein are made. The smallest virus coded polypeptide, the non-structural protein (NS), is made in only small amounts in the coupled system although it is a major virus coded product of infected cells early in infection.


[Klüver-Bucy syndrome and primitive motor reflexes after heavy brain trauma (author's transl)]. A 16-year-old physically and mentally retarded male developed a complete Klüver-Bucy syndrome similar to that demonstrated in animal experiments and characterized by so-called psychic blindness, oral tendencies, hypermetamorphosis, emotional disturbance, and hypersexuality after severe brain trauma with loss of consciousness and a transitory hemiplegia. In addition, transitory oral and grasp reflexes were observed. Despite the severity of the clinical symptoms, the patient was able to realize and report on his subjective experience of the reflex and on the instinct-motivated behavior associated with the Klüver-Bucy syndrome. In the case described, two basic behavior patterns could be differentiated, each influencing drive and the abnormally increased sexuality. The first phase of the illness was dominated by relatively constant automatic reactions. These had the character of pre-determined instinctive behavior and were more or less emotionally neutral. In the second phase of the illness, the instinctive behavior became less evident and the patient was able to respond more normally to his sensory-motor impulses. Also, emotional experience such as frustration, pleasure, and displeasure regained their meaning for the patient.


The use of marsupial x eutherian somatic cell hybrids to study marsupial cell surface antigens. Buck and Bodmer (1976) have developed a technique for identifying an antigen on the surface of human x mouse somatic cell hybrids, specified by a gene on a particular human chromosome. We have successfully adapted this technique to a study of marsupial cell surface antigens. Somatic cell hybrids between Macropus rufus (Marsupialia) lymphocytes and the mouse cell lines PG19 and 1R were injected intraperitoneally into mice of the same inbred strain from which the above cell lines were derived (C57B16J and C3H, respectively). The only identified M. rufus chromosome present in the hybrid cells was the X chromosome. The antisera, after adsorption with PG19 or 1R, were tested using indirect immunofluorescence, against the hybrid cells, and also against sub-clones (derived from hybrids) which had apparently lost the M. rufus X chromosome, or at least its long arm. The results of these tests showed that the absorbed antisera contained reactivity against an M. rufus cell surface antigen (or antigens). The reactions of one of the antisera were most simply interpreted by supposing that it was detecting an M. rufus X-lined antigen(s).


The early development of white adipose tissue. Effects of litter size on the lipoprotein lipase activity of four adipose-tissue depots, serum immunoreactive insulin and tissue cellularity during the first four weeks of life in the rat. 1. Newborn rats were reared in litters of either four or sixteen individuals. The animals from the small litters gained body weight more rapidly than those from large litters during the first 29 days of postnatal life studied. 2. The relative weights of the perigenital, perirenal, subcutaneous and intramuscular white-adipose-tissue sites in the animals from small litters indicated their relative obesity compared with controls. 3. The adipose depots from animals reared in small litters had a greater proportion of lipid present, by weight, and had a greater number of larger fat-cells present in them compared with the depots of animals reared in large litters. 4. Compared with both normal-sized litter controls and animals reared in sixteens, during the period of study the animals from small litters were hypertriacylglycerolaemic but normocholesterolaemic. 5. During suckling the blood glucose concentrations of animals reared in fours were increased, as were the concentrations of circulating immunoreactive insulin. 6. During the 29 days of life studied, in general, the lipoprotein lipase activity of adipose depots from animals reared in fours was greater than for animals in large litters when expressed as mumol of nonesterified fatty acid released from the substrate/h per g fresh weight of tissue, per depot, or per million fat-cells, but were similar per cm(2) of fat-cell surface area. 7. The previously noted [Cryer & Jones (1978) Biochem. J.172, 319-325] pattern of mid-suckling elevation, late-suckling decline and post-weaning increase in the lipoprotein lipase activity of the four white-adipose depots studied was not obliterated by the nutritional manipulations employed. 8. The relation of the enzyme-activity changes and their hormonal stimuli to triacylglycerol accumulation in fat-cells of animals from large and small litters is discussed in relation to the possible significance they may have to our understanding of neonatally induced obesity.


Relationship of free fatty acids and magnesium in ethanol withdrawal in dogs. A group of dogs was conditioned to drink 4-6 g ethanol/kg/day for long periods. Ethanol ingestion was interrupted at monthly intervals in order to study some metabolic changes of the withdrawal period. Plasma long chain free fatty acids (FFA) increased by a maximum mean of 1.4 meq/liter (threefold), and magnesium (Mg) decreased by a maximum mean of 0.4 meq/liter (75% of control during the first 24 hr of simple ethanol withdrawal. Because magnesium salts of FFA are very insoluble, these divergent changes of Mg and FFA suggest that lipolysis may be responsible for the hypomagnesemia that occurs in ethanol withdrawal. In order to control lipolysis, glucose-insulin, glucose alone, and fructose alone were given intravenously for a 4-hr period beginning 14 hr after withdrawal. FFA fell by a maximum of 65% or 0.44 meq/liter and Mg fell by a maximum of 0.31 meq/liter during the glucose-insulin infusions. Nicotinic acid (10-20 mg/kg) in saline produced an identical drop in FFA and a slight rise in Mg. After stopping the nicotinic acid infusion, a sharp rebound rise in FFA and a sharp fall in Mg occurred similar to the simple withdrawal experiments. The sharp divergent changes in FFA and Mg after cessation of nicotinic acid infusion support the prime role of FFA-effecting movements of Mg and the thesis that FFA bind Mg. Control of lipolysis is theoretically sound in therapy of the ethanol withdrawal syndrome.


Electrophysiological studies on embryonic heart cells in culture. Scorpion toxin as a tool to reveal latent fast sodium channel. Trypsin-dispersed heart cells were obtained from 11-day-old chick embryos. After culture as unstirred suspensions in dimethylsulfoxide-containing medium, spherical aggregates of cells beating spontaneously and apparently synchronously for months were obtained. Two kinds of cell were characterized by electrophysiological recordings: (1) cells with a slow rate of depolarizing phase showing tetrodotoxin-resistant action potential and blocked by D 600 ('slow' cells); (2) cells with high value of rising phase which was strongly decreased by tetrodotoxin and in which D 600 provoked uncoupling of excitation-contraction ('fast' cells). Toxin II from Androctonus australis scorpion venom increased the duration of action potential, which was ascribed to a slowing down of Na+ current inactivation and enhance the maximum rate of depolarization, especially in slow cells. Effects were antagonized by tetrodotoxin in both fast and slow cells. Washing experiments confirmed the results of previous studies, namely that tetrodotoxin and scorpion toxin bind to different receptors. It is concluded that slow cells with tetrodotoxin-resistant action potential contain latent fast Na+ channels that are revealed (activated) by toxin binding to the membrane.


The effect of charged synthetic polymers on proteoglycan synthesis and sequestration in chick embryo fibroblast cultures. The polycation, poly(L-lysine), repressed the synthesis of glycosaminoglycans in secondary cultures of chick embryo skin fibroblasts and caused sequestration of glycosaminoglycans around the cells. The synthesis of chondroitin sulphate, dermatan sulphate, hyaluronic acid and a fourth component, thought to be heparan sulphate, were all inhibited to the same extent but the sequestration of the sulphated polymers were greater than that of the unsulphated. The sequestered material was retained around and not within the cells. Incubations with the polyanion, poly(L-glutamate), showed a slight stimulation of glycosaminoglycan synthesis and in these and control incubations (no additions to medium), most of the glycosaminoglycan synthesised appeared in the culture medium. The subsequent addition of poly(L-glutamate) to incubations containing poly(L-lysine) reversed the inhibitory and sequestering effect of the polycation. It was concluded that the inhibition of synthesis by poly(L-lysine) was either a direct effect of poly(L-lysine) on the cell membrane or a result of the high local pericellular concentration of sequestered proteoglycan.


Uptake of fatty acids by cultured cardiac cells from chick embryo: evidence for a facilitation process without energy dependence. Cultured heart cells from chick embryo accumulate fatty acids up to 50 fold at a steady-state level under defined conditions [ref.1]. Studies of fatty acid uptake as a function of different cellular parameters (intracellular ATP and pH, membrane potential and electrochemical gradients for monovalent and divalent cations) show the lack of effect of these factors. The rate of uptake is temperature-dependent. The maximum velocity V is affected with no change in the Km value of the saturable component; the activation energies were found to be 35.5 kJ.mol-1 for palmitate and 42 kJ.mol-1 for oleate. The results are in favour of a facilitation process which leads to an accumulation of fatty acids without energy dependence. The accumulation of fatty acids could be due to their association to intracellular membraneous and/or cytosolic components.


[Parental resistance of irradiated mice (CBA X M523)F1 to lymphocytes: fate of transplanted cells, duration of resistance and its specificity]. The immune response of (CBA X M523)F1 mouse lymphocytes to the antigen (sheep red blood cells) in CBA mice after lethal irradiation was studied. When irradiation, cell transfer and antigen test-injection were performed at the same day the graft activity was inhibited, in comparison with syngeneic system. The activity of donor cells restored on increasing the interval between these processes up to 3 days. Retransplantation of the spleen cells from recipients to irradiated CBA and F1 mice revealed viability of transplanted cells and lack of their readaptation to insyngeneic microenvironment. The recipient's resistance could be specifically overcome by the preinjection of mouse cells combined with cyclophosphamide or without it. The results obtained allow a conclusion that genetic parental resistance of CBA mice to F1 mouse cells is due to the recipient immunocompetent cells which are inactivated 3 days following irradiation. They do not produce cytotoxic effects with respect to donor cells, blocking, however, their activity for some time.


[Diurnal rhythm of changes in the DNA-synthesizing cell count and the mitotic activity of the cells of jejunal epithelium under conditions of subdiaphragmatic vagotomy]. Bilateral subdiaphragmatic vagotomy in rats 7 days after surgery results in increasing proliferative activity of the jejunum epithelium not changing circadian rhythm of the mitotic process. Vagotomy induces changes in the diurnal rhythm structure of DNA-synthesis in the jejunum epithelial cells, i.e. monoapical rhythm in the norm becomes biapical.


Experimental homocystinemia, endothelial lesions and thrombosis. A single intravenous injection of homocystine led to an increase in the number of circulating endothelial cells in the blood of rats. We observed also increased permeability of the lung capillaries, platelet sequestration and activation of the venostatic thrombosis. Endothelial injury is probably the key mechanism of thrombotic and atherosclerotic complications in homocystinuria, an inborn error of the amino acid metabolism.


Manipulations of dietary tryptophan: effects on mouse killing and brain serotonin in the rat. Maintaining rats on a tryptophan-free diet for 4--6 days induced mouse killing in non-killer rats, and significantly facilitated killing in killer rats, as indicated by shorter latencies to kill the mice. The killing responses were similar in topography to the natural killing responses. These changes in killing behavior did not appear to be due to generalized changes in irritability. The increased killing after maintenance on a tryptophan-free diet was accompanied by a 26% reduction in brain serotonin (5-HT) and a 29% reduction in brain 5-hydroxyindoleacetic acid (5-HIAA). When the tryptophan-free diet was supplemented with L-tryptophan (0.5 or 2%), brain 5-HT and 5-HIAA were increased above control levels, and the rat's killing response appeared normal both in terms of latency and topography, similar to that seen in control chow fed animals. While rats consumed less of the tryptophan-free and tryptophan supplemented diets, control subjects deprived of chow such that they lost as much weight as rats fed the tryptophan-free diet, did not show changes in killing behavior. These results are consistent with the hypothesis that central serotonergic systems exert inhibitory control over mouse killing behavior in rats.


Healing of trabeculectomies in rabbits.

Healing of trabeculectomies in rabbits. We studied the healing of trabeculectomies in rabbits with photographs, fluorescein filtration and histological sections. Initially all except 1 developed proven filtration. However, vigorous fibroblastic proliferation (and collagen production) from the subconjunctiva, uvea and sclera combined to close off the fistula by day 14. Groups of eyes treated with 8 mg of prednisolone injected subconjunctivally at operation showed slightly delayed wound healing but by day 30 there was little difference between treated and untreated groups. The steroid treated eyes had complications ranging from endophthalmitis in 1 eye to tissue necrosis in 3 eyes.


Anaerobic mediastinitis and septic shock secondary to esophageal perforation. The authors report unusual complications arising from the ingestion of a small fish bone by a 68-year-old man. These included mediastinitis, empyema, pericarditis and septic shock, probably secondary to a small perforation of the esophagus. After appropriate surgical drainage, antibiotic therapy and supportive therapy the patient made a good recovery.


Autoimmune disorders complicating adolescent Hodgkin's disease. Four adolescents with Hodgkin's disease also developed autoimmune diseases. There were two idiopathic thrombocytopenic purpura (ITP), one polymyositis, and one scleroderma. The first two patients developed ITP in the absence of a spleen, and with their Hodgkin's disease in remission. The first patient with Hodgkin's disease has been continuously free of cancer for over five years. The second patient was a 17-year-old male whoe Hodgkin's disease recurred, but whose disease was in remission at the time the ITP occurred. The polymyositis occurred in an 18-year-old youth when he was in his initial remission for his Hodgkin's disease, but his disease subsequently recurred two years later. This youth presented with Coombs positive autoimmune hemolytic anemia. The polymyositis did not respond to therapy, and he is left with severe muscle wasting and weakness; however, the polymyositis is now quiescent. The scleroderma occurred in an 18-year-old female who had been continuously free of Hodgkin's disease for eight years. The scleroderma did not respond to drug therapy and she now has moderate skin changes, but remains in continuous remission of her Hodgkin's disease. Although there are a few reports of Hodgkin's disease and concurrent autoimmune disorders, physicians dealing with cancer in adolescents should be aware of this association.


Cellular binding proteins for vitamin A in the normal human uterine cervix and in dysplasias. Cellular retinol-binding protein (CRBP) and cellular retinoic acid-binding protein are present in the cytosol of normal human uterine cervical tissues, as detected by ultracentrifugation analysis. Both binding proteins have characteristically high specificity for their respective ligands. In sucrose gradients, both proteins sediment in the 2S region and are of similar molecular weight (M.W. approximately 14,000). In blind analyses of cervical biopsies, obtained under direct vision by colposcopy of normal women (control) or from patients histopathologically diagnosed to have dysplasias or carcinoma in situ (study group), CRBP was not detectable by sucrose gradient analysis in 78.8% of the 33 abnormal biopsies, compared to 23.5% of the 34 controls. This difference was statistically significant (p less than 0.005). In biopsies in which CRBP was detected, the mean levels were 2.76 and 0.72 pmol/mg protein in the cytosol for the control and study groups, respectively. In some subjects from each group, cellular retinoic acid-binding protein but not CRBP was detected in the biopsied tissue. The presence and role of these binding proteins in vitamin A metabolism, epithelial maturation and differentiation in cervical dysplasias, and in situ lesions remain to be investigated.


Efficacy of 6-diazo-5-oxo-L-norleucine and N-[N-gamma-glutamyl-6-diazo-5-oxo-norleucinyl]-6-diazo-5-oxo-norleucine against experimental tumors in conventional and nude mice. The chemotherapeutic effects of 6-diazo-5-oxo-L-norleucine (DON) and N-[N-gamma-glutamyl-6-diazo-5-oxo-norleucinyl]6-diazo-5-oxo-norleucine (azotomycin) were evaluated in a spectrum of transplantable experimental tumor systems including xenografts of human tumors in athymic mice. Both drugs displayed remarkable activity against the murine leukemia L1210 and P388, the Colon Tumors C26 and C38 and the CD8F1 mammary tumor. No significant activity was observed against Lewis lung carcinoma, B16 carcinoma, B16 melanoma, and intracranial ependymoblastoma. DON and azotomycin also exhibited striking inhibitory effects on the growth of s.c. human tumor (MX-1 mammary, LX-1 lung and CX-1 and CX-2 colon) xenografts in athymic mice. With the exception of one colon xenograft (CX-1), all tumor lines were markedly responsive to both drugs. Tumor regressions below the initial tumor sizes of 100 to 300 mg, albeit temporary, were brought about by one course of treatment every 4 days for 3 doses (at optimal dose) with either drug. Although these drugs have been tested previously in the clinic and have shown only limited therapeutic effectiveness, they seem to worthy of a second and closer look in light of the recent laboratory results.


Granule secretion by the luteal cell of the sheep: the fate of the granule membrane. The ultrastructure of the corpus luteum of the sheep has been examined at the mid-stage of the estrous cycle when progesterone secretion is active. Secretory granules are associated with the secretion of this hormone, and the evidence indicates that the granule membrane becomes incorporated into the plasma membrane during exocytosis. Further evidence of this process has been obtained from studies on the uptake of horseradish peroxidase by the luteal cells.


Idiopathic hypertrophic subaortic stenosis and ventricular preexcitation. Two patients who had idiopathic hypertrophic subaortic stenosis (IHSS) and type A ventricular preexcitation were studied and showed variations of the subvalvular flow gradients. The increase in subvalvular gradient, occurring at a time when preexcitation developed, was associated with significant increase of the systolic murmur and the systolic anterior motion of the mitral valve. In patient 1, the significant increase in subvalvular gradient during ventricular preexcitation was also confirmed by catheterization of the left side of the heart. The increase of the gradient with the development of the ventricular preexcitation was apparently due to decreased end-diastolic volume. The importance of ventricular volume considered as a variable affecting outflow tract gradient in idopathic hypertrophic subaortic stenosis is emphasized.


Nucleus-anchoring cytoskeleton in chicken red blood cells. Cytoskeleton of chicken erythrocytes was studies studied after extraction of the cells with Triton X-100. In phase contrast microscopy the extracted cells were seen as ghost-like structures with preserved morphology, distinct nucleus and surrounding plasma membrane remnant. In electron microscopy, dense matrix-like nucleus, fibrillar plasma membrane residue and filaments, ca. 10 nm in diameter, traversing the cytoplasmic domain, were seen. Distinct bands of molecular weight 68000, 53000, 32000 and bands of both higher and lower molecular weight were obtained by polyacrylamide gel electrophoresis of the extracted cells. These results indicate that intermediate filaments, forming the nucleus-anchoring cytoskeleton, are present in nucleated chicken erythrocytes as part of cellular cytoskeleton.


Mitochondrial division in non nucleated sea urchin eggs. An electron microscopic analysis of non nucleated and parthenogenetically activated sea urchin eggs shows figures that can be interpreted as dividing mitochondria. This is in agreement with our previous observations showing that the egg nucleus negatively controls mitochondrial DNA synthesis.


[Diagnostic problems in poisoning with the agaric Cortinarius orellanus Fr. having a nephrotoxic effect]. The authors present a case of fungus poisoning involving nephrotoxic effects. Diagnosis was derived from the typical course of the conditions as well as from the postmortem and mycological investigations of intestinal contents. This is the first case of death due to this kind of poisoning ever described in Czechoslovakia.


Separation of bound from free hormone in radioimmunoassay of lutropin and follitropin. A new technique in which polyethylene glycol is added to double-antibody assays of lutropin and follitropin eliminates the need for an incubation period during the separatory phase. After the antigen-first-antibody equilibrium reaction, the second antibody and polyethylene glycol (30 g/L final dilution) are added, the tubes are immediately centrifuged, each supernate is aspirated, and the precipitated antibody-bound hormone is counted. With this combined polyethylene glycol-second-antibody separatory method, the incubation time is shorter than the usual double-antibody separation procedure, but avoids the problem of high nonspecific precipitation, which occurs with polyethylene glycol alone. In addition, the combined separation technique allows the use of sera containing low titers of second antibody and use of smaller volumes of sera containing high titers of second antibodies, thus conserving second-antibody reagent.


Enzymic assay for serum theophylline.

Enzymic assay for serum theophylline. We describe an original procedure for determination of theophylline in serum. The drug is extracted from 0.4 mL of serum at pH 7.4 with chloroform/isopropanol (20/1 by vol) and back-extracted into sodium hydroxide (1 mmol/L). The inhibition of beef-liver alkaline phosphatases by theophylline in this alkaline phase is measured at 25 degrees C, with p-nitrophenyl phosphate as substrate, in 2-amino-2-methyl-1-propanol buffer, pH 9.4- The reciprocal of enzyme activity and theophylline concentration are linearly related in the range 2 to 60 mg/L. The maximum interference to be expected from 3-methylxanthine would increase apparent theophylline concentration by no more than 1 mg/L. The method is accurate, free of interference by other xanthines and often-coadministered drugs, and results correlate well with those by the immunoenzymic assay. Major advantages are reagent stability, low cost, and simplicity of instrumentation.


Creatine kinase isoenzyme--antibody reactions in immuno-inhibition and immunonephelometry. I have raised, in rabbits, antibodies against MM and BB isoenzymes of creatine kinase. The antibodies produced were homogeneous by Ouchterlony double immunodiffusion and did not cross react with their opposite antigens. Both antisera, however, appeared to be mixtures of antibodies, displaying different equivalences for activation, inactivation, and, possibly, precipitation. Inactivation studies indicated the presence of antibodies effective against dimer only and antibodies effective against monomer or dimer. Both antisera cross reacted with MB and displayed antibodies that appeared to block only half of the activity as well as antibodies that blocked all of the activity. The antisera produced were useful for measuring MB by both immuno-inhibition and immunonephelometry, but neither appears to be advantageous over current electrophoresis or ion-exchange methods. A comparison of decay of MB in patients between activity and mass measurements indicated that activity decay is about 12-fold faster than mass decay.


Functional significance of antibody formation after long-term salmon calcitonin therapy. Twenty patients with Paget's disease were challenged with porcine (PCT) and salmon (SCT) calcitonin before and after 6 months SCT therapy. Acute hypocalcaemia was clearly related to the prevailing rate of bone turnover but responses to PCT showed that treatment had not disturbed this relationship. Although antibodies to SCT developed in eight patients during the period of treatment the acute hypocalcaemic response to SCT was not diminished in relation to bone turnover. It is suggested that antibodies which develop during the course of SCT therapy do not necessarily exert a functionally effective neutralizing action.


Inter- and intrasubject variation in diazepam free fraction. The extent of intersubject variation in diazepam free fraction was measured in fasting plasma of 74 unrelated subjects. Free fraction differences between subjects were significant and ranged from 0.97% to 1.99%. Diazepam free fraction in 29 males was normally distributed about a mean of 1.25% (range, 1.05% to 1.47%), but the distribution in females was skewed to higher free fractions and 40% had values above the highest in males. Albumin concentration (r = -0.27, p less than 0.002) and age (r = 0.44, p less than 0.001) only accounted for a small part of the variation. Within-pair variances were not greater in 11 dizygotic than in 18 monozygotic twin pairs, indicating a greater contribution of environmental than of genetic factors to diazepam binding. The prehemodialysis free fractions of diazepam in 9 uremic patients ranged from 3.44% to 6.69%, and decreased (p less than 0.005) in 7 after 6 hr of hemodialysis. In 10 subjects determination of intrasubject variation in diazepam free fraction between 14-hr fasting and 2-hr postprandial plasma samples indicated that because subjects differ in their pattern of change in free fraction (p less than 0.001), the overall decrease in mean free fraction did not achieve statistical significance (p = 0.10). The mean relative percent change in free fraction within subjects after feeding was 15.2%.


DNA measurements of mitotic chromosomes of the rabbit (Orytolagus cuniculus). DNA cytophotometry was used to obtain the relative DNA content of mitotic chromosomes of Orytolagus cuniculus. DNA ranking of the rabbit chromosomes is presented, and related to the quinacrine and Giemsa-banded karyotype. In addition, the relative DNA content of the short and long arms of each chromosome was calculated and the DNA-based centromeric index, (i.e., the ratio of long-arm DNA to total chromosomal DNA) determined. The Y chromosome is clearly a submetacentric chromosome on the basis of the DNA measurements.


Echocardiography in the diagnosis of idiopathic hypertrophic subaortic stenosis coexisting with pericardial effusion. Of 89 patients with idiopathic hypertrophic subaortic stenosis who had M-mode echocardiagrams recorded, seven patients with coexisting moderate to large pericardial effusions were identified. The clinical profile, M-mode echocardiograms, and cardiac catheterization and angiocardiographic studies in two of the seven patients were analyzed. That the two entities were not associated was suggested by the identification of an etiology for the pericardial effusion in four of the patients. Although the "swinging heart" phenomenon was exhibited in the echocardiograms of each patient, the presence of a significant pericardial effusion did not preclude the ability to establish a diagnosis of idiopathic hypertrophic subaortic stenosis by M-mode echocardiography.


Angiosarcoma of the heart. Diagnosis by echocardiography. Angiosarcoma of the heart is a rare tumor infrequently diagnosed ante mortem. The clinical presentation is usually that of pericarditis or congestive heart failure. This case report illustrates the contribution of echocardiography in suggesting the diagnosis.


Cerebrospinal fluid pathways in a murine mutant. The hy-3/hy-3 mouse is a model of congenital hydrocephalus. The development of the subarachnoid space and extracellular space were studied in normal and hydrocephalic animals. A previous space in the mesenchyme is altered to form the subarachnoid space. Changes in the extracellular space demonstrate that this space is dynamic and acts to compensate for events occurring in the ventricular system. The aqueduct and central spinal canal are also involved in the hydrocephalic process. Change in cerebral vasculature may indicate the processes which lead to irreversibility.


[Glucose-homeostasis in resection and non-resection procedures of the stomach]. Antrectomy or truncal vagotomy with pyloroplasty induces an abnormal alimentary hyperglycemia after ingestion of oral glucose. Low doses of secretin normalize the glucose homeostasis in dogs after antrectomy and truncal vagotomy.


[Healing process of unpreserved homologous articular cartilage transplantation in animal experiments]. Our experimental work was concerned with the healing process of fresh homologous osteochondral grafts in the knee joints of 45 sheep. The ultrastructural investigations pointed clearly to the fact that the cells of fresh homografts completely survive under optimal conditions. Neither transformation nor metaplasia were evident.


[Studies on organ-related postoperative utilization of administered amino acids]. The exclusive postoperative supply of carbohydrates resulted in a significant improvement (24%) of cumulative nitrogen balance, because increased insulin production apparently caused augmented recycling of amino acids liberated during catabolism. Through addition of amino acids the decreased branched chain amino acids returned to normal levels, whereas the significant fall in plasma proteins with short half-life remained unchanged. Therefore it is assumed that the available amino acids (64% of supply) have predominantly anabolic functions in peripheral tissues and fewer are used to synthesize plasma proteins in the liver.


Discordance for achalasia in identical twins. Twenty-year-old twin sisters, believed monozygotic on the basis of extensive blood grouping and cytogenetic studies, are reported. One twin demonstrated classical clinical, radiologic, and manometric features of achalasia, while similar studies in her sister documented perfectly normal esophageal motor function. Genetically determined damage to the esophageal parasympathetic innervation is, therefore, not a likely cause for the esophageal motor dysfunction in achalasia.


[Treatment of diabetes during pregnancy with Des-Phe insulin: preliminary results (author's transl)]. Seven pregnant diabetics were treated with Des-Phe insulin (Hoe 01-S and Hoe 02-S). Daily blood glucose profiles were obtained for 7 to 32 weeks, a total of 4182 measurements. The blood glucose level was well controlled with the insulin preparation. Postprandial blood-glucose increase was counteracted so that even one hour after food the preprandial blood sugar concentration had been reached (Des-Phe insulin Hoe 01-S). This ensured normal carbohydrate metabolism during pregnancy. There were no side effects or skin allergies. The pregnancy proceeded uneventfully in all patients resulting in seven normal infants.


Studies on pituitary follitropin. IV. A conformation specific radioimmunoassay for the ovine hormone. An antiserum to partially purified ovine follitropin (50 x NIH-FSH-S10) shows species specificity. It is conformation dependent and requires the proper recombination of the alpha and beta subunits for maximal reactivity. The isolated alpha subunit is essentially inactive and the hormone specific beta subunit is weakly reactive. The homologous radioimmunoassay is valuable for estimating native ovine follitropin in the presence of free subunits. It also provides a sensitive method to study association-dissociation and structure-function relationships.


Reversal by insulin of concanavalin A inhibition of myotube formation and evidence for common binding sites. Concanavalin A (Con A) inhibits fusion of trypsin-treated myoblasts. This inhibition is reversed by the addition of supraphysiological concentrations (4 micrograms/ml) of insulin either during continuous presence in culture or by pulse additions at 36 and 48 h of culture, just before the time that cultures not treated with Con A undergo myoblast fusion. This reversal is not due to the mitogenic effects of insulin. Under reversal conditions, no specific displacement of bound [125I]iodo-Con A was detected nor did insulin stimulate metabolite uptake. Cell surface replicas of hemocyanin-tagged Con A showed that insulin reversal of the inhibition of myotube formation correlated with the alteration of Con A-binding sites from a clustered configuration present in the inhibited cells to a dispersed state correlated with normal myotube formation. Although a causal relationship has yet to be shown, the data suggest that insulin-mediated reversal of Con A inhibition of myoblast fusion may be related to the ability of insulin at supraphysiological levels to alter the translational mobility of cell surface components containing glucose and/or mannose residues capable of binding Con A. Evidence is presented which suggests that insulin and Con A share common binding sites, since in the physiological range of insulin concentrations (1 ng/ml), Con A pretreatments results in an inhibition of specific [125I]iodo-insulin binding, and antagonistic interactions of insulin and Con A on metabolite uptake and cell proliferation occur. Thus, it appears that the insulin receptors of developing skeletal muscle are glycoproteins containing glycopyranosides.


Properties of the methane mono-oxygenase from extracts of Methylosinus trichosporium OB3b and evidence for its similarity to the enzyme from Methylococcus capsulatus (Bath). 1. The methane mono-oxygenase from Methylosinus trichosporium OB3b was soluble. The only suitable electron donor was NAD(P)H, neither sodium L-ascorbate nor electrons derived from the oxidation of methanol could substitute for NAD(P)H. Evidence is presented for the existence of an NAD+-linked formaldehyde dehydrogenase. 2. Mono-oxygenase activity was not inhibited by a range of potential inhibitors including potassium cyanide, amytal, carbon monoxide or various metal-chelating agents, although 8-hydroxyquinoline and ethyne were effective in this respect. 3. Although the enzyme preparations were unstable on storage, the crude extract could be resolved into two components by ion-exchange chromatography. Activity could be restored to one of the components on addition of purified components from Methylococcus capsulatus (Bath). 4. Cross-reactivity of mono-oxygenase components and other similarities between the enzymes from M. trichosporium and M. capsulatus are discussed. The properties of the M. trichosporium methane mono-oxygenase reported here are contrasted with the properties of the same enzyme reported by others.


Nucleotide sequence coding for the N-terminal region of the matrix protein influenza virus. After polyadenylation in vitro of the influenza virus RNA segment which contains the coding information for the matrix protein, a cDNA copy can be made using the primer p(dT)8-dA and reverse transcriptase. The sequence of 166 nucleotides of the cDNA was determined by a modification [Brownlee, G. G. & Cartwright, E. M. (1977) J. Mol. Biol, 114, 93--117] of the plus/minus method [Sanger, F. & Coulson, A. R. (1975) J. Mol. Biol. 94, 441--481] and adaptation of the "dideoxy" method [Sanger, F., Nicklen, S. & Coulson, A. R. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 5463--5467] for sequencing DNA. The cDNA sequences is of the same sense as the mRNA for matrix protein and contains a potential initiating codon, d(ATG), at position 26--28. When matrix protein purified from virus particles was digested with chymotrypsin or trypsin and the amino acid compositions of separated peptides determined, one peptide containing nine amino acids found which had a composition corresponding to that predicted by the cDNA sequence following the first methionine codon, confirming that protein synthesis initiates at this position. The compositions of four other peptides matches those predicted from the nucleotide sequence. There is no processing of the N terminus of the protein before incorporation into the virus particle except for removal of the N-terminal methionine and addition of a "blocking" group on the resulting N-terminal serine residue.


Presynaptic subsensitivity as a possible basis for sensitization by long-term dopamine mimetics. A possible cellular basis for dopaminergic sensitization by long-term dopamine mimetics was examined in rat brain striatum. Long-term apomorphine or amphetamine administration (10 mg/kg/day for 14 days) resulted in a decrease in the specific binding of 3H-apomorphine, but no change in 3H-haloperidol binding. Long-term apomorphine treatment also enhanced the cataleptogenic action of haloperidol, with many rats being spontaneously cataleptic after apomorphine withdrawal. It is suggested that the reduced 3H-apomorphine binding signifies less presynaptic receptors. This permits less autoregulation and enhanced dopamine agonist action, possibly accounting for the dopaminergic sensitization by long-term agonists.


The effect of an intrauterine device on myometrial glycogen in the hamster. The myometrical glycogen content of the uterine horns of cycling, castrated and castrated hormone-treated hamsters was basically unaffected by the presence of an intrauterine device (IUD). The study also indicates that visual evaluation of histochemical localization of glycogen is not adequate to determine minor differences in staining intensity. The "estrogen-like" effect or stimulatory effect of the IUD on uterine glycogen in the rat is not duplicated in the hamster.


[Dynamics of hormone secretion during chronic emotional stress]. Study of spontaneous secretion of corticosteroids and thyroid hormones and the direct hormonal response to stress revealed the pathogenic effect of chronic combined emotional stress upon the hormonal function of adrenal glands. The hippocampus takes part in formation of the emotional tension in response to stress stimulus and of the following hormonal secretion.


The effect of polyanion on the enzymatic degradation of modified deoxyribonucleoprotein samples. The degradation of DNA in modified deoxyribonucleoprotein samples by DNase II in the presence of a polyanion (polypentose sulphate) was studied. The behaviour of DNP samples partially depleted of histone and treated with acetic anhydride is characterized by an increased accessibility of their DNA to DNase II, by a decreased polyanion amount needed for the maximum enzymatic degradation of their DNA, and, finally, by a more profound inhibitory effect of higher polyanion amounts on the DNase II-catalyzed degradation. On the other hand, the behaviour of DNP samples treated with formaldehyde is characterized by a decreased accessibility of their DNA to DNase II, by a limited increase in the enzymatic degradation of their DNA in the presence of the polyanion, and finally by a less profound inhibitory effect of higher polyanion amounts on the DNase II-catalyzed degradation.


[Effect of pellet size on respiratory activity and motility of cryopreserved spermatocytes]. The effect of different pellet volumes at freeze preservation in human semen was studied. It was measured the motility of spermatozoa during an incubation time of several hours at 37 degrees C and the alteration of oxygen consumption after adding rotenone, succinate, digitonine or 2,4-dinitrophenol. - The motility of cryopreserved spermatozoa does not show significant differences after preservation procedure. In respect of oxygen consumption the smallest differences from native material were obtained at a pellet volume of 0,2 to 0,3 cm3. Explanations for this behaviour are discussed.


Effect of neonatal stunting on development of rats: large litter rearing. As nurslings, rat pups reared in large litters showed reduced frequencies of returns to their nest from other parts of the home cage and reached maximum levels of nest returns at older ages than control animals from small litters. These differences were not due to differences in activity level or attraction to the nest but appeared to be perceptual in nature. As adults, animals from large litters showed deficits in learning the 1st 2 problems in a series of visual discriminations in a modified version of the Lashley jumping stand technique in which food reinforcement was not used. The deficits were confined to male animals only and were the result of stronger position habits in the early part of training, rather than an inability to make the discriminations. A comparison of these results with those obtained when other methods of stunting animals are employed revealed that different methods of stunting may result in both common and divergent effects on behavior. One long-term consequence of large litter rearing appears to be increased emotional response to noval situations.


Three parameters affecting interlitter variations. We studied 3 parameters affecting average body weight differences between mouse litters and have arrived at an approximation of the contribution that each makes to such differences. These parameters are birth weight, genetically based growth potential variations, and maternal competence. We have developed methods for controlling these parameters and show that through their application we can reduce litter-average body weight differences from our normal value of more than 24% to about 3%. The vanished 21% consists of 12% due to differences in maternal competence, 6% due to growth potential differences, and 3% due to initial weight differences.


Argon vs. neodymium YAG laser photocoagulation of experimental canine gastric ulcers. A neodymium YAG (Nd:YAG) laser was evaluated in a dog ulcer model used in the same manner as is recommended for bleeding patients (power 55 W, divergence angle 4 degrees, with CO2 gas-jet assistance). The experiments were performed during sterile laparotomy in heparinized dogs. Bleeding gastric ulcers were photocoagulated until bleeding stopped and then examined histologically 7 days later when depth of tissue injury was maximal. In the first series of experiments, the Nd:YAG laser was compared with the 7-W argon laser in the same dogs. Both lasers stopped bleeding from all experimental ulcers. The 55-W Nd:YAG laser caused full-thickness injury to the gastric wall beneath 11 of the 14 treated ulcers, whereas the 7-W argon laser caused no full-thickness injury beneath 14 treated ulcers. In a second series of experiments, we tried to determine whether varying exposure times with the 55-W Nd:YAG laser would make it less injurious; it did not. In a third series of experiments, the 55-W Nd:YAG laser was tested with and without CO2 gas-jet assistance in order to determine if this would affect the depth of injury; it did not. In the final series of experiments, the wattage of the Nd:YAG laser was varied to see if this would reduce depth of injury; lower wattage did not stop bleeding, and intermediate and higher wattages did stop bleeding but did not reduce depth of injury. We conclude that the 55-W Nd:YAG laser as it is currently used clinically produces deeper tissue damage than the argon laser in our animal model. This damage is not reduced by changes in power, duration of exposure, or the presence of gas-jet assistance.


Inhibition of hepatic regeneration in rats by acute and chronic ethanol intoxication. We studied the effects of acute and chronic ethanol feeding on hepatic regeneration in rats after partial hepatectomy and toxic liver injury produced by D-galactosamine. Ethanol, when administered as a single dose (6 g/kg), inhibited 3H-thymidine incorporation into hepatic DNA; this effect depended in part on the time of ethanol feeding after partial hepatectomy. Multiple ethanol feedings produced an even greater inhibition, which persisted for at least 48 hr after partial hepatectomy. Rats chronically fed ethanol for 30 days also failed to achieve a hepatic proliferative response to either partial hepatectomy or D-galactosamine-induced hepatitis, comparable with isocaloric pair-fed controls. These investigations suggest that there may be a certain metabolic state in the hepatocyte cell cycle that is most susceptible to the action(s) of ethanol; inhibition of liver regeneration by acute or chronic ethanol consumption may result in delayed recovery from prior or coincident liver injury.


Nematocide effect of entomophilic and entomophagous fungi. Study of nematocide effect of 48 species of entomophilic and entomophagous fungi from the class Fungi imperfecti against Panagrellus redivivus and Rhabditis oxycerca nematodes revealed a high frequency of fungi with nematocide effect. A close correlation between these two effects was disclosed by measuring the intensity of the nematocide and insecticide effect of fungal extracts.


Personality traits of 47,XYY and 47,XXY males found among juvenile delinquents. The mental features of nine 47,XYY males and six 47,XXY males found among Japanese juvenile delinquents were examined. The results of intelligence tests suggested a slight impairment among 47,XYY and 47,XXY males. The psychiatric interviews and the psychological tests revealed that 47,XYY males had a high activity level while 47,XXY males had a low one. Analyses of the offense type and psychophysiological features supported these findings. As the activity level is considered to be largely determined genetically, the extra Y is suspected to be responsible for the high activity level and the extra X in males to be responsible for the low activity level.


Combination of differential sister chromatid staining, G-banding pattern, and X-inactivation pattern. A method is presented for obtaining a combination of differential sister chromatid staining. G-banding and X-chromosome inactivation pattern. The result of this method enables a precise localization of the sister chromatid exchange points to particular bands of individual chromosomes.


X-linked mental retardation with macro-orchidism and the fragile site at Xq 27 or 28. Data are presented suggesting that the form of X-linked mental retardation with macro-orchidism and the form associated with a fragile site at Xq27 or 28 are the same entity.


Regulation of radiation pollution: its possible usefulness in strategy for intervention against chemical mutagens. Over the last 25 years, a large amount of work has been carried out in the field of safety with regard to ionizing radiations. This has resulted in the establishment of norms of security which have been accepted in most countries. These norms are not perfect; they are constantly reviewed and, from time to time, revised; but, such as they are, they are of tremendous usefulness. These standards may serve as a model of what must be achieved in the field of pollution by chemical mutagens. Further, the norms already in force for radiation may serve as a reference for the establishment of similar norms for the major environmental mutagens. It is not possible to assign a specific factor of equivalence with radiation (rad-equivalence) to a given chemical pollutant; but it is possible to establish point-equivalences with regard to well-defined biological systems under well-defined conditions. When they are accurately determined, such point values may be useful and sensible. In this area, as with radiation, it is impossible to find perfect solutions; however, approximate solutions can be arrived at that would already represent a reat progress with regard to the present situation, and which could later be improved as work in the field allows.


Effects of antimicrotubule agents, potassium and inhibitors of energy production on hCG secretion. The BeWo trophoblastic cell line was employed to assess the requirement for microtubules and cellular energy in human chorionic gonadotropin (hCG) secretion. In contrast to the general inhibitory effect of colchicine and vincristine on hormone secretion in systems involving exocytosis, wide concentration ranges of these antimicrotubule agents caused enhancement of hCG secretion. Similarly, cytochalasin B, an agent that interferes with microfilament function, doubled both basal hCG secretion, and secretion of hCG stimulated by 1 mM dibutyryl cyclic AMP plus 1 mM theophylline (dbT). Inhibitors of cellular energy production (2,4-dinitrophenol, malonate, azide) decreased both secreted and cellular levels of hormone. High concentrations of K+ gave no enhancement of basal or dbT-stimulated hCG secretion, nor any reduction of cellular hCG levels. These findings contrasted with observations of others in secretory systems involving exocytosis, in which high K+ potentiated basal or stimulated hormone release and depleted cellular stores of hormone. It was concluded that the process of hCG secretion in the malignant trophoblast is fundamentally different from the mechanism of protein hormone secretion in other tissues.


[Comparative studies on the use of equimolar 14C-labeled glucose and fructose infusions in normal subjects and patients with liver cirrhosis]. In 11 normal subjects (NS) and 12 patients with liver cirrhosis (LC) the utilisation of 14C-glucose and 14C-fructose infusions (0.75 g/kg/h for 4 h) was compared. There were nor relevant side effects. Lactate and pyruvate were in both groups during fructose infusion slightly increased compared to glucose infusion. The free fatty acids were significantly decreased. The serum glucose level rose more in LC than in NS when given fructose infusion. During glucose and fructose infusion in LC higher insulin concentrations were calculated than in NS. 15 min after infusion of 14C-fructose 20% of the total serum activity was 14C-glucose, after 2 to 4 h the level was 30%. Differences between NS and LC were not found to be significant. The specific activity of 14CO2 was the same in both the 14C-glucose infusion and the 14C-fructose infusion. The glucose oxidation was impaired in LC, but not the 14CO2-exhalation during infusion of 14C-fructose. Unimpaired 14CO2-exhalation, and normal utilisation and conversion to glucose are arguments for the use of fructose in infusion treatment of cirrhotics.


[Effect of a 3.6 o/o dextran Ringer acetate solution on moderately hypovolemic patients in intensive care]. The effects of 1000 ml 3.6% dextran Ringer's acetate solution (Mw 61,800; infusion time 60 min) on circulation, blood and plasma volume, blood viscosity and renal function parameters were investigated, the renal elimination of dextran also being measured. During the entire investigation period no significant alterations of cardiocirculatory parameters, blood and plasma volume and rheological properties of blood could be demonstrated, the same holding true for kidney function values. Dextran excretion was about 33% three hours after the end of infusion, reaching 51% of the whole amount given 21 hours later. It has to be assumed that mainly macromolecular dextran components of less efficiency are retained intravascularly. Taking into consideration the practically absent volume expanding effect - even at the end of infusion period - this dextran solution cannot be recommended as a blood substitute during acute hemorrhage or as an "expander" in other shock states.


[Behavior of the serum fatty acid patterns during administration of an MCT-containing fat emulsion]. After a continuous intravenous 3-hour infusion of 500 ml fat emulsion containing MCT, composed of 25 g soybean oil and 25 g MCT oils, there was an infusion-induced increase of all fatty acids administered. The medium-chain fatty acids (C8-C12) revealed ideal steady-state conditions and were quickly eliminated after the infusion. The reaction of linoleic acid (C18:2) was not as favorable. Linoleic acid increased steadily during the infusion, and its elimination rate from the serum was delayed.


[The effect of formula diets with a modified fat content on human plasma phosphatide fractions]. Variation of human plasma phospholipids by formula diets with different composition of fat are caused by changes of the lecithin fraction, which amounts to 70 to 80% of total phospholipids. The changes of lysolecithin correspond to that of lecithin. The absolute amounts of sphingomyelin are unchanged by the different formula diets, while the relative amounts are negatively correlated to the lecithin fraction.


Intramuscular inoculation of live respiratory syncytial virus induces immunity in cotton rats. Intramuscular inoculation of infant or weanling cotton rats with 10(2.2) to 10(4) plaque-forming units of respiratory syncytial virus induced significant or complete resistance to infection in both the upper and lower portions of the respiratory tract. This resistance did not appear to be the result of in vitro neutralization of virus during homogenization of tissue. Virus was not recovered from the local site of inoculation after 5 min and was never detected in the respiratory tract of intramuscularly inoculated rats. Attempts to detect viral antigens at the site of inoculation by using indirect immunofluorescence were unsuccessful. However, inactivation of infectivity of three different strains of respiratory syncytial virus markedly reduced or completely ablated antigenicity and protective efficacy by the intramuscular route. This suggests that these viruses underwent limited replication, perhaps restricted to an abortive cycle, at the local site of inoculation. An immunosuppressive effect of passive maternally derived immunity was observed. Only 50% of weanling rats possessing passive maternal serum antibody were successfully immunized by intramuscular vaccination with live virus.


Splenic influence on the development of a local pulmonary immune response. The role of the spleen in the development of specific antibody-forming cells (sAFC) in the pulmonary draining lymph nodes (pdLNC) of hamsters after local inoculation of sheep erythrocytes (SRBC) was evaluated. The role of the spleen was viewed from two vantage points. Panels of animals were either splenectomized with appropriate sham-operated controls before intratracheal inoculation of SRBC, or panels were immunized intravenously simultaneously with the local inoculation of antigen. The presence of an intact spleen was not necessary for the induction of a sAFC response to occur in the pdLNC. Similar numbers of immunoglobulin M (IgM) sAFC were recorded in the pdLNC on day 4 of both sham-operated and splenectomized animals. However, an enhancement of this local response occurred on day 7 if the animals were systemically immunized and therefore demonstrated active participation of the spleen in the specific immune response. The results support the hypothesis that although a local response may occur in the pdLFC in the absence of a spleen or a splenic response, the presence of a systemic or splenic response appears to be important for the enhancement of local IgM sAFC response. These observations suggest that the immune defenses involved in the lower respiratory tract may differ from those in upper respiratory tract and other mucosally lined organs in that the response of the spleen to the antigen affects the local response to that antigen.


Comparison of intratracheal and intravenous inoculation of sheep erythrocytes in the induction of local and systemic immune responses. The interdependence of the local and systemic immune systems in the development of the immune responses relating to the lung was evaluated. Hamsters were inoculated with 10(9) sheep erythrocytes (SRBC) via local (intratracheal) or systemic (intravenous) routes of immunization. The local immune response was quantitated by the specific antibody-forming cell (sAFC) response in the pulmonary draining lymph nodes (pdLNC). sAFC in the spleens and serum hemagglutination titers evaluated the systemic immune response. The local inoculation of antigen was superior for induction of the maximal numbers of immunoglobulin M (IgM) and non-IgM sAFC in the pdLNC at 4 days post-immunization. However, the local response coould be enhanced by a concomitant splenic response. The local route of inoculation did not consistently induce an sAFC response in the spleen, but when the spleen was involved, two unique observations were recorded regarding the IgM response: (i) an approximate twofold increase in the numbers of sAFC was seen in the pdLNC on day 7 post-inoculation in animals that had a splenic sAFC response; and (ii) the appearance of an early (day 4) serum IgM hemagglutination titer was observed in the animals with a splenic sAFC response. However, the appearance of IgG serum hemagglutination titers was independent of the indirect sAFC response in the spleen but correlated with the appearance of indirect sAFC response in the pdLNC. As expected, intravenous inoculation of antigen was superior for inducing a systemic IgM response. We concluded that the local and systemic immune responses are related and interdependent in providing immune reactivity related to the lung. Whereas the systemic response enhanced both the serum IgM titer and the specific IgM antibody-forming cell response in the draining nodes, we observed that the local response was sufficient for the IgG serum hemagglutination titer even in the absence of a splenic sAFC response.


Effect of immune status on dengue 2 virus replication in cultured leukocytes from infants and children. Cord blood leukocytes from neonates with maternal dengue antibody supported dengue 2 virus replication in vitro; those from neonates without maternal antibody did not. Cord bloods of infants born to dengue-immune mothers contained a potent enhancing factor which gradually decayed with age and which was absent from neonates born to nonimmune mothers. Permissiveness of cultures of washed peripheral blood leukocytes from infants with maternal antibody declined steadily with increasing age in parallel with the decay of maternal antibody, and the leukocytes were no longer permissive after 10 to 12 months. The demonstration of a dengue maternal infection-enhancing factor in human cord blood from dengue-immune mothers supports the hypothesis that severe primary dengue hemorrhagic fever with shock seen in Bangkok infants is related to maternal immune status.


Adherence of human peripheral blood lymphocytes to measles-virus infected cells: modulation by solubilized rhesus erythrocyte membranes and carbohydrates. The adherence of human peripheral blood lymphocytes to HeLa cells persistently infected with measles virus (HeLa-K11) was studied. The following data were observed. (i) The proportion of HeLa-K11 cells with adherent human peripheral blood lymphocytes of rhesus monkey erythrocytes was similar over a wide range of ratios of HeLa-K11 cells to lymphocytes or erythrocytes. (ii) The great majority of human peripheral blood lymphocytes and erythrocytes reacted with the same HeLa-K11 cell (iii). The adherence of lymphocytes or erythrocytes to HeLa-K11 cells was blocked by rabbit anti-measles virus antibody or solubilized monkey erythrocyte membranes. The pretreatment of erythrocytes or lymphocytes with receptor-destroying enzyme did not alter their adherence properties. (iv) The pattern of inhibition observed with several carbohydrates was similar in both the erythrocyte and the lymphocyte adherence assays. These data are consistent with the possibility that the receptor present on both rhesus monkey erythrocytes and human lymphocytes has similar specificities and biochemical composition.


Partial purification and characterization of two Cladosporium herbarum allergens. Two important Cladosporium herbarum allergens (Ag-54 and Ag-32) were partly purified by means of diafiltration, gel filtration and isoelectric focusing. Modifications of quantitative immunological techniques, including autoradiography, were used as assay systems for allergenic activity and to provide a basis for evaluation of the immunological purity of the allergens. Ag-54 appeared to have a molecular weight of approximately 25,000 daltons and an isoelectric point of 5.0. The molecular weight of Ag-32 appeared to be approximately 13,000 daltons. It consisted of 5 molecular species differing in isoelectric points from 3.4 to 4.4 in intervals of approximately 0.25 pH units. These variants of Ag-32 seemed to be antigenically and allergenically identical.


Serum amyloid protein SAA, C-reactive protein and lysozyme in leprosy. Serum amyloid protein (SAA) appears to be the precursor of amyloid protein AA, the non-immunoglobulin fibril protein of secondary amyloidosis. Since amyloidosis is known to occur in high frequency associated with lepromatous leprosy (LL), we have examined the SAA levels in untreated LL patients and compared them to the levels observed in patients with tuberculoid leprosy (TT) and a large number observed in healthy controls. We found that SAA is markedly elevated in LL when compared to TT and controls. No clear correlation could be established with C-reactive protein, a well-documented acute phase reactant, or serum lysozyme levels that reflect the presence of monocyte activity. This study showed that SAA levels in leprosy do not appear to be a reflection of inflammatory activity or monocyte turnover. Whether amyloidosis will be more prevalent in patients who have higher SAA levels remains to be determined.


Lucio's phenomenon: a comparative histological study. To study further the pathogenesis of Lucio's phenomenon, we have made a comparative histological study of 11 patients with Lucio's phenomenon and 12 with ENL. Confirming the findings of others, Lucio's reaction could be distinguished from ENL by epidermal necrosis and by necrotizing vasculitis manifesting necrosis in the walls of superficial vessels and severe, focal endothelial proliferation of mid-dermal vessels. Furthermore, in Lucio's phenomenon large numbers of AFB were found in evidently normal and in swollen or proliferating endothelial cells. We hypothesize that patients with Lucio's phenomenon have an exceptionally deficient defense mechanism, allowing unrestricted proliferation of AFB in endothelial cells, facilitating contact between bacterial antigen and circulating antibody and leading to infarction; also, this nadir of resistance allows unimpeded dissemination of AFB, accounting for the clinical features of diffuse non-nodular leprosy. Thus, an explanation is offered for the restriction of Lucio's phenomenon to patients with diffuse non-nodular lepromatous leprosy.


In vivo effect of DDS on phytohemagglutinin (PHA)-induced lymphocyte transformation cultures in normal healthy volunteers. Depression in PHA-induced lymphocyte transformation in peripheral blood has been observed in 15 healthy volunteers after administration of DDS (100 mg/day) for seven days. In vitro culture of lymphocytes obtained from these volunteers in DDS-free normal AB serum has not altered the blast cell numbers. Lymphocytes of these volunteers have been found to contain a significant amount of DDS, ranging from 0.42 to 3.2 micrograms per 10.6 lymphocytes.


Unusual electrocardiographic presentation of idiopathic hypertrophic subaortic stenosis. A patient with idiopathic hypertrophic subaortic stenosis and normal coronary arteries, with dynamic electrocardiographic changes resembling acute myocardial ischemia, is presented. A definite association between the electrocardiographic changes and the idiopathic hypertrophic subaortic stenosis cannot be absolutely confirmed; however, the findings demonstrate the broad spectrum of electrocardiographic presentation in this disease.


Studies on relationships between the structure and the glycosidase-inhibiting activity of HPAAO and its analogs. p-Hydroxyphenylacetaldoxime (HPAAO) (anti and syn forms) obtained by fermentation and its analogs chemically synthesized were tested for their activities to inhibit various glycosidases. HPAAO inhibited bovine liver beta-galactosidase in a competitive manner at pH 7.0 with an apparent Ki value of 8 x 10(-8) M. HPAAO also inhibited various mammalian beta-glycosidases which had pH optima between 6.0 and 8.0. The syn form of HPAAO was found to be more active than the anti form against bovine liver neutral beta-galactosidase. It was concluded that the oxime moiety of HPAAO and its analogs was essential for their enzyme-inhibiting activity and the activities of aromatic or aliphatic oxime derivatives were dependent on the number of carbon atoms in their alkyl-chains.


A cytochemical and ultrastructural study of the "S.I.F." cells in cat sympathetic ganglia. According to the hypothesis of Eccles and Libet, the small intensely fluorescent cells (S.I.F. cells) in the sympathetic ganglion would represent an essential element in the inhibition of the principal neuron. As a contribution to the study of this important problem, we have investigated serial sections in superior cervical (S.C.G.) and celiac (C.G.) ganglia of the cat, a species that has not been extensively studied up to now, both by fluorescence and electron microscopy. We have shown that the "S.I.F." cells are three times fewer in the cat S.C.G. than in the rat S.C.G. There are five times more "S.I.F." cells in the C.G. of the cat than in the S.C.G. of the same species. Moreover we have described two types of "S.I.F." cells. Type I is composed of cells characterized by highly polymorphous large dense-cored vesicles. These cells lack processes and are grouped in clusters centered on fenestrated capillaries. They could be endocrine function cells. Type II is formed of isolated cells which exibit long processes and establish synaptic junctions with the dendrites of the principal neurons. In this case, the dense-cored vesicles are very regular and much smaller. These cells could be equivalent to interneurons. Type I very strongly predominates in the S.C.G. and C.G. of the cat where it represents more than 90% of the "S.I.F." cell total observed by fluorescence microscopy. A priori such a quantitative and qualitative heterogeneity hardly consistent with Eccles and Libet's hypothesis based on the existence of dopaminergic interneurons only, allows the question to be raised as to the functional significance of the "S.I.F." cells in ganglion physiology. The notion of modulation of ganglionic transmission does not seem to be quiered by these new data but could be founded on different forms of action embodied in the broader conception of the neuromodulation phenomenon.


Formation of molecules of biological interest from formaldehyde and hydroxylamine in a modified sea medium. As reported previously, glycine, serine, aspartic acid, and beta-alanine are the predominant amino acids produced from equimolecular formaldehyde and hydroxylamine in a modified sea medium enriched with essential transition metal ions. The time course of formation of the amino acids, and related substances was studied. Among the amino acids, glycine was produced earlier. Urea was produced initially and disappeared rapidly. Formation of cyanide, glycinenitrile, and glycinamide preceded the formation of amino acids. Probable pathways for the formation of amino acids and urea are suggested.


Preparation of ruthenium(II) and ruthenium(III) myoglobin and the reaction of dioxygen, and carbon monoxide, with ruthenium(II) myoglobin. Ruthenium myoglobins have been prepared by the reconstitution of horse heart apomyoglobin with either ruthenium(II) or ruthenium(III) mesoporphyrin IX (MpIX) derivatives. The ruthenium(II) and -(III) myo globins (RuMb and RuMb+, respectively) contain one ruthenium porphyrin/heme binding site; the species are readily interconverted using dithionite for reduction and bromine for oxidation. RuMb binds carbon monoxide to give the known carbonyl complex. Reversible oxygenation occurs readily with protein-free RuII(MpIX) species in dimethylformamide, but RuMb in phosphate buffer is irreversibly oxidized by dioxygen to give RuMb+ via an outer sphere electron transfer mechanism.


Intermediate (skeletin) filaments in heart Purkinje fibers. A correlative morphological and biochemical identification with evidence of a cytoskeletal function. Cow Purkinje fibers contain a population of free cytoplasmic filaments which consistently differ in ultrastructural appearance from actin and myosin filaments, irrespective of preparation technique. The fixation and staining techniques, however, influenced the filament diameter, which was found to be 7.4--9.5 nm for filaments in plastic-embedded material, and 7.0 nm in cryo-sectioned material, thus intermediate as compared to actin and myosin filaments. Cross-sectional profiles suggested that the intermediate-sized filaments are composed of four subfilaments. To provide a basis for further biochemical investigations on the filaments, extraction procedures were carried out to remove other cell organelles. Electron microscopy showed that undulating bundles of intermediate filaments converging towards desmosomes still remained, after the extractions, together with Z-disk material. In spite of the extensive extraction, the shape of the individual cells and the assemblies of cell bundles remained intact. This confirms that the intermediate filaments of cow Purkinje fibers together with desmosomes do in fact have a cytoskeletal function. On account of (a) the cytoskeletal function of the filaments, (b) the similarities to the smooth muscle "100-A filament" protein subunit skeletin, and (c) the inadequate and confusing existing terminology, we suggest that the filaments be named "skeletin filaments."


Nucleation of polar actin filament assembly by a positively charged surface. Polylysine-coated polystyrene beads can nucleate polar assembly of monomeric actin into filamentous form. This nucleation has been demonstrated by a combination of biochemical and structural experiments. The polylysine-coated beads accelerate the rate of actin assembly as detected by two different biochemical assays. Subsequent examination of the beads by electron microscopy reveals numerous actin filaments of similar length radiating from the beads. ATP promotes this bead-induced acceleration of assembly. Decoration of the filaments with the myosin fragment S1 shows that these filaments all have the same polarity, with the arrowhead pattern pointing toward the bead. The relevance of the system to in vitro mechanisms and its usefulness in other studies are discussed.


Role of spindle microtubules in the control of cell cycle timing. Sea urchin eggs are used to investigate the involvement of spindle microtubules in the mechanisms that control the timing of cell cycle events. Eggs are treated for 4 min with Colcemid at prophase of the first mitosis. No microtubules are assembled for at least 3 h, and the eggs do not divide. These eggs show repeated cycles of nuclear envelope breakdown (NEB) and nuclear envelope reformation (NER). Mitosis (NEB to NER) is twice as long in Colcemid-treated eggs as in the untreated controls. Interphase (NER to NEB) is the same in both. Thus, each cycle is prolonged entirely in mitosis. The chromosomes of treated eggs condense and eventually split into separate chromatids which do not move apart. This "canaphase" splitting is substantially delayed relative to anaphase onset in the control eggs. Treated eggs are irradiated after NEB with 366-nm light to inactivate the Colcemid. This allows the eggs to assemble normal spindles and divide. Up to 14 min after NEB, delays in the start of microtubule assembly give equal delays in anaphase onset, cleavage, and the events of the following cell cycle. Regardless of the delay, anaphase follows irradiation by the normal prometaphase duration. The quantity of spindle microtubules also influences the timing of mitotic events. Short Colcemid treatments administered in prophase of second division cause eggs to assemble small spindles. One blastomere is irradiated after NEB to provide a control cell with a normal-sized spindle. Cells with diminished spindles always initiate anaphase later than their controls. Telophase events are correspondingly delayed. This work demonstrates that spindle microtubules are involved in the mechanisms that control the time when the cell will initiate anaphase, finish mitosis, and start the next cell cycle.


Actin microfilaments, cell shape, and secretory processes in isolated rat hepatocytes. Effect of phalloidin and cytochalasin D. The effects of phalloidin and cytochalasin D, drugs which, respectively, stabilize and destabilize actin microfilaments, have been tested on isolated rat hepatocytes. Both drugs produced a modification of cell shape, characterized by protrusions bulging from the cytoplasm. In phalloidin-treated hepatocytes, an accumulation of actin microfilamentous network was detectable at the base of each protrusion by electron microscopy, immunofluorescence, and HMM decoration. This accumulation of microfilaments was absent in cytochalasin D-treated cells. The release of triglycerides, an index of very low density lipoprotein secretion, was inhibited by phalloidin or cytochalasin D, and accompanied by an increase in cellular triglycerides. At the electron microscope examination, triglyceride accumulation was represented by fat droplets and vesicle-enclosed, very low density lipoprotein-like particles. Total protein and albumin secretion was only very slightly modified by either one of these drugs. With the use of various phalloidin analogs, a correlation was observed between their respective ability to stabilize F-actin in vitro, and their effects on cell shape and triglyceride secretion. In conclusion, phalloidin, and cytochalasin D: (a) modify the shape of isolated hepatocytes; (b) inhibit lipoprotein secretion. These effects possibly result from a modification of actin microfilament function.