The use of althesin in drug-resistent status epilepticus. The properties of Althesin (anticonvulsant activity, depression of oxygen consumption, lowering of ICP, rapid excretion) led us to use this steroid combination to treat 11 patients in status epilepticus resistant to the standard drugs (benzodiazepines and barbiturates). The administration of Althesin by slow intravenous injection was ineffective in 2 of the 3 patients thus treated. The doses used (2--10 ml) were probably too small. One only administration of a 10% solution of Althesin in 10% fructose by intravenous drip (the rate was calculated so as to obtain the burst suppression stage at the EEG) stopped status epilepticus in 7 of the 9 patients thus treated. In this group the doses used varied from 25 to 50 ml. The 2 patients in whom it was necessary to repeat Althesin administration and combine it with other drugs had both been operated on for severe brain injuries involving marked cerebral edema. In spite of the very small number of cases, the definitive arrest of status epilepticus obtained in 8 out of 11 patients first treated with other drugs is encouraging: Althesin probably may be regarded as an adjunct in the treatment of status epilepticus.


Causes of death in institutionalized epileptics. To assess the causes of death in institutionalized epileptics, the patient records, death certificates, and other data from the only hospital for epileptics in Finland (Vaajasalo Hospital) were reexamined. During the years 1900--1976, 179 inpatients in Vaajasalo Hospital died; this was 12% of all inpatients. The most common causes of death were as follows: pneumonia in 40 cases, seizures in 34 cases (single seizure in 18 and status epileptics in 16), drowning in 29 cases, stroke in 10 cases, and heart infarct in 9 cases. Chronic intoxication caused by phenytoin and/or phenobarbitol was a common supplementary factor leading to death in patients who died of pneumonia or seizures. Thirteen deaths were recorded as suicides or suspected suicides (11 by drowning and 2 by strangulation). The results reflect the severe epilepsy and the poor conditions of the patients, as well as the poor condition of the local facilities in the past. The information obtained should be useful in the effort to improve medical care for these patients.


Hemisphere-dependent cognitive performances in epileptic patients. Short-term memory (STM), long-term memory (LTM), and recency judgment were investigated in 75 right-handed nonaphasic patients without visual deficits who had epileptic foci localized in one of four different regions of the brain: right temporal lobe, left temporal lobe, right frontal lobe, and left frontal lobe. The results led to the following conclusions: (1) left temporal lesions yielded comparatively greater impairment for the verbal LTM task, whereas right temporal lesions were more disruptive for the spatial LTM task; (2) no significant differences between left- and right-brain-damaged patients in STM tasks were obtained; (3) left frontal lesions yielded deficits in a recency task in accordance with hemispheric specialization; (4) only prolonged epileptic syndromes impaired neuropsychological performance in those tests; (5) comparisons between male and female patients and between those with and without clear evidence of a structural lesion did not reach statistical significance.


Distinction between normal and epileptic rhythms in rodent sensorimotor cortex. Characteristics of spontaneous spindling in electrocorticograms from awake rodents are described. Comparisons are made between this spontaneous activity and epileptic activity generated in the sensorimotor cortex.


Do anticonvulsants reduce the efficacy of oral contraceptives? Women in our area with epilepsy who were also taking oral contraceptives were identified. Of 82 patients taking oral contraceptives, 41 had used both anticonvulsants and oral contraceptives for a total of 955 months. Three documented oral contraceptive (pill) failures occurred during this period, whereas the expected number of 0.12 (relative risk, 25; 95% confience interval, 5 to 73). No pill failures were observed in 2,278 months among women with epilepsy who were taking oral contraceptives but who were not taking anticonvulsants at this time. Thus our data support the suggestion that there is an increased rate of pill failure among women taking anticonvulsants. In view of this diminished effectiveness, the advisability of using oral contraceptives rather than one of the other forms of contraception when anticonvulsant medication is being used concurrently may need to be reevaluated.


Ictal body scheme disturbance induced by looking through a small opening. The case of a patient is reported in which seizures consisting of a feeling that the left arm was absent were induced by looking through a small opening (e.g. camera view finder). The EEG showed right parietal sharp wave discharges between and during such seizures. This observation is interpreted as an example of a reflex epilepsy triggered by the specific stimulus of looking through a small opening, and involving the right parietal lobe.


Pseudohypoparathyroidism and epilepsy: diagnostic value of computerized cranial tomography. Computerized cranial tomograms (CCTs) unexpectedly showed bilateral symmetrical calcifications in the basal ganglia and frontal areas in two unrelated epileptic patients 12 and 13 years of age. The patients presented with a variety of seizures, some with focal features; these seizures were resistant to medication in the first case. Subsequent testing revealed hypocalcemia and other biochemical and radiologic features of pseudohypoparathyroidism, despite absence of the usual phenotypic features, tetanic symptoms, and positive family history. The CCT scan may provide the first clue to an underlying hypocalcemic disorder in an epileptic patient even when the skull X-ray is normal. Early detection of this metabolic condition by CCT scanning allows specific treatment to restore serum calcium levels to normal, which usually eliminates seizures and favors optimal cerebral functioning. Serial CCT scanning also provides a useful means for following the intracerebral calcifications, which remained unchanged after 1 and 2 years of normocalcemia in our 2 patients.


Effectiveness of clonazepan in startle-induced seizures. Five cerebral-palsied children and adolescents with severe startle epilepsy became seizure-free after clonazepan was introduced into their existing anticonvulsant drug regimens. The drug was withdrawn in one case because of side effects. Two hemiparetic patients who had startle epilepsy as the only epileptic manifestation remained permanently controlled after a mean of 34 months of continuous therapy. Reappearance of startle-induced seizures occurred after 1 and 4 years in two other patients with the Lennox-Gastaut syndrome. A possible explanation for the effectiveness of clonazepan in this form of reflex epilepsy may involve inhibition of brainstem mechanisms mediating pathologically enhanced reactions in these patients, thus avoiding secondary activation of a discharging focus in the vicinity of the motor supplementary area.


The use of ionising radiation for the treatment of injuries to flexor tendons and supporting ligaments in horses. A technique was developed using radioactive isotopes as a source of radiation for the treatment of injuries to the superficial and deep flexor tendons and the associated ligaments in the horse. The treatment area was sub-divided so that different dosages could be applied over the limb as necessary. A plaster of Paris impression was taken of the whole area to be treated. In the isotope laboratory a plaster negative was made and loaded with the dose of radioactive isotope. The loaded cast was then strapped to the horse's limb for the calculated time, usually about three days. A total of 42 horses were treated and follow up information was obtained from 28. Twenty-five animals raced again; two relapsed before racing and one was destroyed with navicular disease. Ten of the 42 horses had been treated by firing before irradiation. Five of these returned to racing but the history of four of them was not known.


Oxygenation of pulmonary vein blood in conscious and anaesthetised ponies. Catheters were implanted through the left atrium into large veins drawing blood from regions of the diaphragmatic lobes in five ponies. Measurement of oxygen and carbon dioxide tensions in blood samples drawn through these catheters showed that in laterally recumbent conscious and anaesthetised animals the function of the lowermost lung is progressively impaired while in the supine position the function of both lungs is adversely affected.


Enteroliths in horses--a retrospective study of 30 cases. In a retrospective study of selected cases, abdominal colic in 30 horses was attributed to enterolith obstructions of the large intestine. Obstructions caused by "true" enteroliths were confined to horses more than four years old. Prominent clinical features were recurrent mild abdominal pain, inappetance, gaseous distension and minimal intestinal motility. The various aspects of the clinical syndrome, including diagnostic problems and clinical management, are discussed. Most obstructing enteroliths were found near the beginning of the small colon and most horses contained only a single major concretion. Enteroliths were formed by mineral deposition in concentric layers about a central nidus of ingested material and were spherical or tetrahedral in shape. Intestinal concretions were found to consist primarily of ammonium magnesium phosphate.


Staphylococcal septic arthritis in three horses. Three horses were diagnosed as having monarticular septic arthritis due to Staphylococcus aureus on the basis of culture of articular cartilage, synovial membrane and/or synovial fluid. The organisms were all well recognised human phage types and in two cases demonstrated beta-lactamase (penicillinase) activity. Details of case histories are presented and the bacteriological techniques and antibiotic management with cloxacillin, methicillin and penicillin discussed. Following treatment, sterile cultures of synovial fluid were achieved in all cases, but in two horses the infections resulted in degenerative articular changes. This necessitated arthrodesis of the fetlock joint in one case.


A case of carpal intersynovial fistula in a horse. A tentative diagnosis of a fistula between the tendon sheaths of the right extensor carpi radialis and the intercarpal joint was confirmed by positive contrast radiography following injection of sodium diatrizoate into the joint. Surgical removal of a tag of synovial membrane and repair of the fistula was carried out. Post surgical treatment included mild exercise to prevent adhesions within the sheath. A month following discharge, contrast radiography revealed no communication between joint and tendon sheath. At this time mild distension of the sheath was relieved by aspiration and it did not recur when the horse returned to work following a further month of rest.


Purification and some properties of a hitherto-unknown enzyme reducing the carbon-carbon double bond of alpha, beta-unsaturated carboxylate anions. 2-Enoate-reductase, a previously unknown soluble enzyme is present in Clostridium kluyveri and another Clostridium species growing on (E)-2-butenoate. From the latter the reductase was purified 88-fold with an overall yield up to 74%. The enzyme was pure as judged by polyacrylamide gel electrophoresis with and without sodium dodecyl sulphate as well as by isoelectric focusing. The purification of the enzyme was performed in the presence of (E)-2-methyl-2-butenoate as substrate to keep the enzyme in the oxidized state and under anaerobic conditions. The purification procedure included an ammonium sulphate precipitation, chromatography on DEAE-Sepharose CL-6B, hydroxylapatite and Sepharose CL-6B. The enzyme reduces different alpha,beta-unsaturated carboxylate anions such as (E)-2-butenoate, (E)-2-methyl-2-butenoate, (E)-cinnamate and probably many others in a NADH-dependent reaction to the saturated carboxylate anions. Fumarate, 3-phenyl-2-propinate, 2-enoyl-methyl and CoA esters proved not to be substrates for the purified reductase. NADPH does not act as an electron donor. The enzyme was shown to have a molecular weight of about 450,000 by gel chromatography. It consists of subunits with a molecular weight of 78,000. Per subunit about 1 FAD, 3.5--3.8 atoms of iron and 4.0 labile sulphur atoms have been found indicating a conjugated iron-sulphur flavo-protein. Copper could not be detected. The isoelectric point was 8.4. As shown by absorption spectroscopy the enzyme can be reduced by NADH and reoxidized with dichloroindophenol, hexacyanoferrate III, oxygen and substrates. Addition of 8 mol p-hydroxymercuribenzoate to 1 mol subunit completely destroyed the activity of the reductase. So far no physiological role of the enzyme is known.


Hydrodynamic parameters of the detergent-solubilised hydrogenase from Paracoccus denitrificans. The hydrogenase from Paracoccus denitrificans, which is an intrinsic membrane protein, has been solubilised from membranes by Triton X-100. The partial specific volume of the solubilised protein has been determined using sucrose density gradient centrifugation in H2O and 2H2O. The values of the specific volumes of hydrogenase, measured in the presence or absence of Triton X-100, are 0.73 and 0.74 ml . g-1, respectively, indicating that hydrogenase binds much less than one micelle of Triton X-100. The sedimentation coefficient of hydrogenase is increased from 10.4 S to 15.9 S on removal of detergent. The Stokes' radius of hydrogenase, determined by gel filtration on Sepharose 6B, is 5.5 nm in the presence of Triton X-100 compared to 6.7 nm in the absence of detergent. The apparent molecular weight therefore increases from 242,500 to 466,000 on removal of detergent. In the presence of urea and sodium dodecylsulphate, the hydrogenase has an apparent molecular weight of 63,000. The enzyme therefore behaves as a non-covalently linked tetramer in the presence of Triton X-100. Removal of Triton X-100 results in association of tetramers to form octamers.


The kinetics of ATP-dependent exonuclease V from Micrococcus lysodeikticus. A Michaelian dependence on DNA concentration. The ATP-dependent exonuclease V from Micrococcus lysodekticus shows a Michaelian relation between steady-state velocity and the concentration of T7 DNA substrate. The Km (expressed as a mass concentration) does not change when the T7 DNA is broken into smaller fragments by a restriction enzyme. This is interpreted to mean that the predominant process by which the exonuclease-V--DNA complex breaks down is digestion of the entire DNA molecule rather than physical dissociation, in accord with the already known processive nature of degradation by this enzyme. The way that the V and Km towards DNA vary with ATP and ADP concentration suggests that enzyme-DNA complex is predominantly formed by reaction of DNA with an enzyme-ATP complex rather than with bare enzyme.


Regulation of glycogen synthesis in rat-hepatocyte cultures by glucose, insulin and glucocorticoids. The changes in glycogen content and in its rate of synthesis in two-day-old primary cultures of rat hepatocytes were assessed under various conditions. Hepatocytes cultivated in serum-free and hormone-free medium switch from glycogen degradation to glycogen deposition at 10.3 mM glucose. After pretreatment of the cells with glucocorticoids this threshold was reduced, in the absence or presence of insulin, to 5.4 or 1.2 mM glucose, respectively. The rate of glycogen synthesis in the presence of 10 mM glucose was amplified from 5 nmol x h-1 x mg protein-1 to 20 nmol glucose x h-1 x mg protein-1 after pretreatment with triamcinolone. Glucagon pretreatment also significantly increased the subsequent glycogen synthesis rate. Insulin addition accelerated glycogen synthesis about twofold regardless of the pretreatment. The dose-response relationship between insulin concentration and glycogen synthesis rate showed half-maximal effect at 0.62 +/- 0.22 nM (mean +/- S.D.) insulin. Pretreatment of hepatocytes with glucocorticoids, glucagon, insulin or combinations of these hormones did not significantly change the concentration which gives the half-maximal effect.


Trans-membrane translocation of proteins. The direct transfer model. As a start towards a deeper understanding of the transmembrane transport of proteins, the transfer of a nascent chain through the lipophilic core of a membrane is discussed from a physico-chemical point of view. Some simple considerations of the energetics of protein structure, together with experimental data on the transfer process, form the basis for a detailed and quantifiable model, accounting for the extrusion of secreted proteins as well as for the insertion of trans-membrane proteins.


Partial purification and characterization of the proteins from the 40-S ribosomes of Artemia salina and wheat germ. The proteins were extracted from purified 40-S ribosomes derived from wheat germ and Artemia salina and separated by carboxymethylcellulose ion-exchange chromatography. Approximately four proteins from Artemia and four proteins from wheat germ were separated in a state of high purity. All proteins were identified by co-electrophoresis using a two-dimensional polyacrylamide gel system. A total of 30 unique proteins were found for Artemia and 32 proteins for wheat. The molecular weights of all proteins were estimated by sodium dodecylsulfate gel electrophoresis. Assuming each protein to be present in one copy per 40-S ribosome, the total protein molecular weight was estimated to be 560,000 associated with Artemia 40-S particles and 550,000 associated with wheat germ 40-S ribosomes.


Effect of sulphated glycopeptides on kinetics of 3-O-methyl glucose and 2-deoxyglucose transport by epithelial cells isolated from rabbit small intestine. Suspensions of cells isolated from rabbit small intestine were prepared which showed a considerable rate of respiration after 4 h of their isolation. Rabbit intestinal cells accumulated 3-O-methyl-glucose by an active transport mechanism which was Na+-dependent whereas 2-deoxyglucose was transported by a saturable pathway. Analysis of the initial transport data indicated that the uptake of both sugars by cells followed Michaelis-Menten-type kinetics. These cells also showed a considerable degree of 2-deoxyglucose phosphorylation and the transport of the sugar into the cells appeared to be a rate-limiting factor for its phosphorylation. Sulphated glycopeptide had no effect on the respiration rate of these cells but it increased the transport of 3-O-methyl-glucose and 2-deoxyglucose into cells by many fold compared with untreated cells. Kinetic analysis of the data indicated that the increase in the transport was due to increased V while the Km remained unchanged. Presence of sulphated glycopeptide had no effect on the kinetics of phosphorylation of 2-deoxyglucose. These effects of sulphated glycopeptide on the transport activity of the cells have been discussed as a result of an increase in the number of sugar sites in the cell membrane due to unmasking or activation of preexisting sites owing to the interaction between the cell membrane components and highly polyanionic sulphated glycopeptides.


N-Acetylaminoacyl-p-nitranilidase from human placenta. Purification and some properties. An enzyme hydrolyzing N-acetylaminoacyl-p-nitroanilides has been isolated from mature human placentae by a six-step procedure comprising extraction from a placenta homogenate, ammonium sulfate fractionation, treatment with isopentyl alcohol, chromatography on CM-Sephadex, protamine sulfate precipitation and gel filtration on an Ultrogel AcA 34 column. About 2500-fold enrichement was achieved from placenta homogenate. The purified enzyme preparation showed a single band on polyacrylamide disc electrophoresis. The molecular weight was estimated to be 380,000 by gel filtration. Placental extracts contain two main isoenzymes of pI 3.9 and 4.5 respectively. Activity was strongly inhibited by chloromercuribenzoate, slightly inhibited by Ca2+ and cysteine; no activation could be detected. The enzyme exhibits an exopeptidase-like activity towards acetyl-dipeptides with a certain specifity towards N-acetylalanyl-alanine; N-acetylalanine-p-nitroanilide, however, is hydrolyzed four times faster. With N-acetylalanine-p-nitroanilide as substrate the pH optimum was 8.0--8.2; Km was 2.13 mmol/l. N-Acetylleucine-p-nitroanilide and N-acetyltyrosine-p-nitroanilide were split slowly; N-acetylalanyl-alanyl-alanine-p-nitroanilide, N-butyloxycarbonyl-alanyl-alanine-p-nitroanilide, unsubstituted analogous aminoacyl-p-nitroanilides and several protein substrates were not hydrolyzed. The functions of the enzyme are still unknown.


Binding of Semliki Forest virus and its spike glycoproteins to cells. We have studied the binding of the Semliki Forest virus and its isolated spike glycoproteins, in the form of water-soluble octameric complexes, to various cells at 5 degrees C. The number of viruses bound per cell increased strongly with increasing free concentrations of virus up to about 0.2 nM. At higher concentrations smaller increases in binding were observed but saturation was not achieved. The number of viruses bound at a given free concentration was widely different for different cells. For some cells the binding of the virus was maximal at pH 6.8 with little decrease at lower pH, for other cells it was maximal around pH 6.0. The spike protein complexes were used at 100 times higher molar concentrations than the virus. The binding increased strongly with increasing free concentrations up to about 50 nM and saturation was obtained at higher concentrations. Up to 1.3 X 10(6) spike protein complexes could be bound per cell but great variation could be seen between different cell types. For all cells maximal binding was found below pH 6.0. Together with earlier observations, our results suggest that the virus can bind to a cell by two different modes. Around neutral pH the virus binds to specific glycoproteins and at low pH unspecifically to the lipids of the plasma membrane. The possible physiological roles of these two types of binding are discussed.


Methyl acceptors for protein methylase II from human-erythrocyte membrane. Membrane proteins from human erythrocytes were methylated with purified protein methylase II (S-adenosylmethionine:protein-carboxyl O-methyltransferase, EC.2.1.1.24). The methylated proteins were analyzed by dodecyl sulfate/polyacrylamide gel electrophoresis. Monomeric and dimeric glycophorin A (NaIO4/Schiff-2 and NaIO4/Schiff-1 positive bands) and 'band 4.5' were identified as two major classes of methyl-acceptor polypeptides for protein methylase II. In rabbit erythrocyte membrane where glycophorin A is absent, 'band 4.5' was the only major methyl-acceptor protein component. Extracted and purified glycophorin A from human erythrocytes was also found to be an excellent substrate for protein methylase II with a Km of 35.7 microM. The role of erythrocyte membrane protein methylation is discussed with regard to membrane function.


Permissive effect of dexamethasone on glucagon induction of urea-cycle enzymes in perifused primary monolayer cultures of rat hepatocytes. Parenchymal cells from adult rat liver, cultured in perifused monolayers, increased the levels of urea-cycle enzymes between 15% and 60% in response to glucagon within 24 h. This stimulation was drastically enhanced by the simultaneous presence of dexamethasone, especially in the case of argininosuccinate synthetase and argininosuccinate lyase, which increased nearly threefold. Dexamethasone itself produced only negligible stimulation, but exerted a similar effect on the stimulatory action of glucagon, if it was exclusively present during 6 h prior to the glucagon treatment, suggesting a permissive action of this hormone. The effect of glucagon, particularly in the presence of dexamethasone, was mimicked by dibutyryl adenosine 3':5'-monophosphate, whereas epinephrine was ineffective. All stimulations induced by hormones or dibutyryl adenosine 3':5'-monophosphate were abolished by cycloheximide, suggesting the involvement of protein synthesis in the induction process. Using the usual culture technique with a discontinuous supply of medium no significant effect of glucagon and dexamethasone could be measured. This striking difference between both culture systems indicates that perifusion is the more adequate in vitro system for studies of the regulation of enzyme levels. Possible reasons for the failure of hormonal stimulation of urea-cycle enzymes in normal monolayer culture are discussed.


Ribosome biosynthesis is not necessary for initiation of DNA replication. Synthesis of mature 28-S ribosomal RNA and 60-S ribosomal subunits is inhibited in baby hamster kidney (BHK) cell line ts 422E at non-permissive temperature (39 degrees C). This leads to a 66% decrease of total ribosomes per cell, a marked imbalance between the large and small ribosomal subunits in the cytoplasm and a decrease of cells per dish after prolonged culture at 30 degrees C. However, inhibition of ribosome synthesis does not affect progression of cells through the G1 period of the cell division cycle, the length of the pre-replicative period, and the rate of entry of cells into S phase. In contrast to culture at non-permissive temperature, culture of BHK ts 422E cells in the presence of 0.04 micrograms/ml actinomycin D at 33 degrees C inhibits markedly the entry into S period. It is concluded that low doses of actinomycin D exert their inhibitory effect on cell growth by preventing maturation and transport of mRNA rather than by interfering with ribosome synthesis. Microfluorometric analysis revealed only slight differences in the distribution of BHK ts 422E cells in G1, S and G2 phases of the cycle either when cultured at 33 degrees C or at 39 degrees C. When too few ribosomes per cell are produced in BHK ts 422E cells at 39 degrees C, cells do not seem to be arrested reversibly at a specific point of the cell cycle but rather to die at random.


Purification and properties of a histone acetyltransferase from Artemia salina, highly efficient with H1 histone. An histone acetyltransferase has been purified from nuclei of 40-h-old Artemia salina larvae. The enzyme is very unstable at 0 degrees C, requires free -SH groups for activity and is rapidly inactivated at 40 degrees C. The optimal pH for activity is 8.5 and the activity is half inhibited by millimolar concentrations of Mn2+, Ca2+ or Mg2+ or decimolar concentrations of Na+ and K+. The molecular weight of the enzyme, determined by gel filtration chromatography, changed with the ionic strength of the medium (280,000 in 10 mM Tris . HCl, 170,000 in 0.2 M KCl). The very-lysine-rich histone H1 is a better substrate acceptor than the arginine-rich histones H3 or H4. Under proper conditions, the enzyme can modify all the internal lysyl residues in histones H1 and H4. The acetylation of H1 is inhibited when all the other histone fractions are present in the assay mixture.


1H-Nuclear magnetic-resonance studies on glycophorin and its carbohydrate-containing tryptic peptides. The proton nuclear magnetic resonance (1H-NMR) spectra of glycophorin and its tryptic sialoglycopeptides were investigated. From the intensities of the assigned resonances it was concluded that all of the residues in the sialoglycopeptides are sufficiently mobile in conformation to give sharp resonances, while in glycophorin this is true for only approximately 80% of the peptide backbone. The resonances of the central sequence of some 20 of the hydrophobic residues are strongly broadened. This region is probably that of alpha-helical structure which is known to aggregate. The linewidths and intensities of the resonances are not, or only slightly, affected by changing the ionic strength, temperature or by carboxymethylation of the Met-81 residue in glycophorin. Glycophorin was found to bind about 100 mol sodium dodecylsulphate/mol protein as derived from studies on linebroadening of the latter's C-3 to C-11 methylene resonances. The bound dodecyl-sulphate probably increases the mobilities of the hydrophobic residues in the protein as these resonance intensities are increased by the binding. The carbohydrate chains in glycophorin were conformationally mobile; no evidence was found for tight carbohydrate-protein interactions. The relevance of flexible carbohydrate chains in membrane glycoproteins is discussed in relation to cell surface chemistry.


Purification, some properties and the complete primary structures of two protease inhibitors (DE-3 and DE-4) from Macrotyloma axillare seed. The Macrotyloma axillare plant, belonging to the Leguminosae family, is a perennial climbing or trailing herb 0.2--3.5 m long. The plant is indigenous to South Africa and it occurs in the warm dry northern parts of the Transvaal. It has been introduced into Australia, where the seed are used as animal food. Two protease inhibitors, DE-3 and DE-4, were purified from Macrotyloma axillare seed by gel filtration on Sephadex G-50 followed by ion-exchange chromatography on DEAE-cellulose. They each comprise 76 amino acid residues including 14 half-cystine residues. The complete primary structures of the two protease innibitors have been elucidated and their sequences are 67% identical. The inhibitor specificities, the sequences, the invariant amino acid residues and the reactive inhibitor sites of protease inhibitors DE-3 and DE-4 resemble the corresponding properties of the Bowman-Birk double-headed protease inhibitor group. The cysteine residues are in similar locations to those in protease inhibitors of known structure so they are presumed to link similarly.


A double-blind pediatric evaluation of levamisole in the prevention of recurrent upper respiratory tract infections. Levamisole was tested double-blind in 106 children with recurrent upper respiratory tract infections. They received either levamisole (n = 53) or placebo (n = 53) 0.5 ml/kg bodyweight b.i.d. for two consecutive days each week for six months. A control examination was performed every two months. Both groups were compared by means of the Fisher-test and the Mann-Whitney U-test (two-tailed probability each). Improvement was observed more frequently in the levamisole group with regard to the number of episodes of infection, and the total duration and severity of the infections. No side-effects, except for some stomach complaints in one levamisole patient, were reported.


Coagulation studies on umbilical arterial and venous blood from normal newborn babies. Coagulation studies using conventional methods and chromogenic substrates were performed on umbilical arterial and venous blood from 33 newborns after delivery. In the arterial samples, thrombin time (TT) was significantly prolonged and the activities of factors I, II, V and VII, as well as the inhibitors heparin, antithrombin III and antiplasmin, were significantly decreased. This could probably be explained by a mild form of disseminated intravascular coagulation (DIC) occurring in the baby during delivery.


Pattern of growth hormone response to insulin, arginine and haemodialysis in uraemic children. Plasma growth hormone (GH) concentrations after insulin and arginine stimulation were estimated in 11 dialyzed and 6 non-dialyzed children with chronic renal failure. Twenty healthy children served as controls. Plasma GH peak concentration and estimation of the total area under the plasma GH concentration-time curve by the trapezoidal rule were used to evaluate results. Elevated basal GH levels and an exaggerated response to the stimuli were seen in several of the patients. The causes of the abnormal GH secretion and the role of high GH levels in carbohydrate intolerance are discussed. No consistent pattern was seen in GH secretion during haemodialysis without glucose in the dialysate. In children undergoing haemodialysis with a fluid containing glucose, plasma GH fell considerably.


Two sisters with isovaleric acidaemia, multiple attacks of ketoacidosis and normal development. Two sisters with isovaleric acidaemia are described. Both had multiple attacks of acetonaemic vomiting, sometimes leading to subcoma. Despite this they showed a completely normal mental development. Biochemical studies, clinical follow-up and attempts at treatment are presented.


Diaphragmatic hernia in a female newborn with focal dermal hypoplasia and marked asymmetric malformations (Goltz-Gorlin syndrome). We report a female newborn with focal dermal hypoplasia (Goltz-Gorlin Syndrome) and marked asymmetric malformations on the right side of the body. Diaphragmatic hernia on the same side, which has not been reported in this syndrome, led to perinatal complications.


Biochemical assessment of thiamine nutrition in childhood. The activity of red cell transketolase and TPP-effect were determined in 1028 infants and children, aged 1 month to 14 years, using a micromethod. All the children were healthy and without evidence of malnutrition. Transketolase activity decreased in the second year of life, followed by a constant increase up to the 6th year. Activity did not change in the next four years, then a steady decrease occurred. TPP-effect was not as stable as transketolase activity, and decreased significantly from years 3 to 5 and increased significantly from years 5 to 7. Beginning with year 10, TPP-effect decreased gradually. 6.6% of all children investigated had marginally deficient TPP-effects and 1.3% were biochemically deficient without clinical signs of avitaminosis. A group of 37 adults had transketolase activities and TPP-effect comparable to those of the 13 year-olds. Effect appeared to be marginal in 5.4% and deficient in 2.7%.


Hypercalciuria in idiopathic Fanconi syndrome. A 9 year old girl with idiopathic Fanconi syndrome and hypercalciuria is described. In order to determine whether the increased calcium excretion was directly or indirectly due to the disturbed phosphate metabolism, the behavior of the calcium excretion during therapy, the serum levels of 1,25-dihydroxyvitamin D and parathyroid hormone, and the effect of parathyroid hormone on the renal tubules were investigated. Normal serum 1,25-dihydroxyvitamin D and parathyroid hormone levels, lack of a correlation between the serum phosphate concentration and the degree of hypercalciuria, as well as unsuccessful therapy of the hypercalciuria with oral phosphate indicate that the increased calcium excretion cannot be explained by impaired renal phosphate reabsorption. The hypercalciuria in the patient was therefore regarded as being due to a primary decrease of tubular calcium reabsorption.


Association between HLA-A1,B8 in children with extrinsic asthma and IgA deficiency. The tissue types, immunoglobulin levels, and the presence of circulating autoantibodies were investigated in 57 children. Fifteen of these children suffered from bronchial asthma and, in addition, had no or very little IgA in their serum and saliva (Group 1 patients). Another fifteen children with asthma but normal immunoglobulin levels in serum and saliva (Group 2 patients), seven patients with selective IgA deficiency but without allergic diseases (Group 3 patients), and twenty healthy children served as controls. Sixty per cent of the Group 1 patients had the phenotype HLA-A1,B8, whereas this tissue type was found only in 27, 14 and 15 per cent, respectively, of the Group 2 and Group 3 patients and the healthy children. Furthermore, high IgM- and IgE levels were observed in most Group 1 patients, and in five of these patients (33 per cent) autoantibodies were present in the serum. In addition, eczema and glomerulonephritis occurred rather frequently in this group of patients. Conversely, normal immunoglobulin levels and absence of circulating autoantibodies were found in the remaining three groups of children. The results emphasize the heterogeneity of the IgA deficiency syndrome, and the question is raised as to whether the tissue type HLA-A1,B8 observed in most Group 1 patients reflects the abnormal immune reactivity of these patients.


Trisomy 9 mosaicism with punctate mineralization in developing cartilages. A case of mosaic trisomy 9 (46,XX/47,XX,+9) is described. The main clinical findings included intrauterine growth retardation, failure to thrive, hydrocephalus, deeply set eyes, prominent antihelix, highly arched palate, subluxation of the interphalangeal joints of some fingers, hip dislocation, excessive sweating, and punctate mineralization in developing cartilages.


A rapid unfavorable outcome of Wegener's granulomatosis in early childhood. Wegener's Granulomatosis was suspected in a 27-month-old female with a nodular, necrotizing lesion of the nose, diffuse subcutaneous nodules, and erythematous desquamation of the entire body. From 20 months of age on she had a purulent nasal discharge, recurrent infections of the upper and lower respiratory tract, a Coombs positive anemia, and enlargement of the spleen and liver. Treatment with azathioprine and corticosteroids produced transient improvement but three months later a dramatic relapse occurred. Cyclophosphamide was substituted for azathioprine but 10 days later the patient died and the autopsy confirmed the diagnosis of Wegener's Granulomatosis. The early age of onset of the disease may explain the unfavorable outcome, despite treatment with cytotoxic agents.


Prognosis of prophylactic anticoagulant treatment in ischaemic stroke. Two groups of patients who had a stroke, prolonged reversible ischaemic neurological deficit or transient ischaemic attacks were compared; one group was treated with anticoagulants (phenprocoumon) and a matched control group was treated with other substances. Results showed a similar survival rate in both groups, but significantly fewer recurrences were seen in the surviving patients treated with anticoagulants.


Zoster meningitis and radiculomeningitis after tick bite. Cytological findings in cerebrospinal fluid. Cytological changes in 9 samples of cerebrospinal fluid (CSF) from 5 patients with zoster meningitis and 12 samples from 5 patients with radiculomeningitis after tick bite were examined. 5 days after the onset of the skin eruption, the CSF cells in zoster meningitis consisted of many mononuclear blast forms--large lymphocytes and 'immature' plasma cells. 11-15 days after the onset of the skin rash, they were replaced by small lymphocytes, some 'mature' plasma cells and monohistiocytes. In radiculomeningitis after tick bite, however, the CSF cells examined 10-21 days after the onset of radicular pain consisted of many large lymphocytes, immature plasma cells and a few neutrophils. From these findings, it might be suggested that the acute meningeal reaction of zoster meningitis subsides within about 1 week after the onset of the skin rash, but that of radiculomeningitis after tick bite continues at least for 2-3 weeks after the onset of radicular pain.


Reciprocal reflex activity in ocular muscles: implications in spontaneous blinking and Bell's phenomenon. Levator palpebrae (m. lev. palp.) and rectus superior (m. rect. sup.) are two synergistic muscles in vertical gaze movements. However, they show an antagonistic behavior during electrically induced blink and glabellar reflexes. This can be considered as an easy and useful method for their differentiation in the electrophysiological study of certain oculopalpebral motor disorders. Blinking consists of a series of reflex activities, the most outstanding ones being an early m. lev. palp. basal activity inhibition, followed by the activation of the orbicularis oculi muscle and a brief increase in m. rect. sup. activity (reflex Bell phenomenon). This coordinated reflex organization could be perfectly related to that known in other human nociceptive reflexes.


A genetic study of febrile convulsions.

A genetic study of febrile convulsions. 307 probands with febrile convulsions classed as the simple type (131 children) and the complicated type (176 children) were genetically analyzed. There was a tendency toward familial aggregation of febrile convulsions, and genetic involvement was suggested. The multifactorial mode of inheritance best agreed with the observations. (1) The ratio of incidence of febrile convulsions in siblings of probands in the present study (19.9%) to the incidence in the general population (2.9%), i.e., 6.85, was rather close to the expected ratio of 5.87 from the multifactorial inheritance system. (2) The incidence in siblings tended to be higher when either or both of their parents had a history of febrile convulsions. In 2-child families where the first child was affected, the incidence in the second child tended to increase in parallel with the increasing incidence in parents. (3) The incidence in siblings was higher if probands were males. (4) The heritability, when estimated by Falconer's procedure, was as high as 76%, showing that febrile convulsions are strongly genetically predisposed. These findings were more distinctly observed in the simple type than in the complicated type.


Normal functional capacity of blood lymphocytes in multiple sclerosis measured by stimulation with mitogens. The effect of phytohemagglutinin (PHA) and pokeweed mitogen (PWM) on blood lymphocytes was studied in multiple sclerosis, patients with acute CNS infections, and healthy controls. No significant differences in lymphocyte proliferation were registered between the group of MS patients and the two other groups, irrespective of mitogen used. The ratio between the PWM and PHA responses did not differ significantly from the ratios in the two other groups. The lymphocyte proliferation with PWM was significantly lower in the group with acute CNS infections compared with controls. The results indicate that alterations in cell-mediated immunity, if present in MS, are not confined to lymphocytes outside the CNS.


A study on Sturge-Weber syndrome. Report of a case associated with infantile spasms and electroencephalographic evolution in five cases. A case of classical Sturge-Weber syndrome associated with infantile spasms (female infant, aged 4 months) was presented. The association of both conditions would be very rare since no similar case was found among 214 cases of Sturge-Weber syndrome and 1,180 cases of infantile spasms gathered from the literature, except for one who was reported by Millichap et al. Evolutional changes of electroencephalographic findings were followed up for about 3 years on average in 5 personal cases of Sturge-Weber syndrome. Unilateral depression of electrical activity in the cerebral hemisphere ipsilateral to the facial nevi was the constant finding. Focal spike discharges were noticed only in the contralateral hemisphere in 3 cases, only in the ipsilateral in 1, and in the bilateral in 1.


Oral zinc sulphate as long-term treatment in Wilson's disease (hepatolenticular degeneration). Clinical amelioration, clearance of Kayser-Fleischer rings and rising of ceruloplasmin concentration are described in a patient with the classical findings of Wilson's disease. These changes occurred during a 14-year period in which he used oral zinc sulphate (three times daily 200 mg) as the only medication to influence copper metabolism. Before starting this long-term zinc sulphate therapy he had used D-penicillamine (three times daily 300 mg) for only 6 weeks. The antagonistic action of zinc sulphate on copper resorption with amelioration of the clinical condition has been described before in this patient in 1961 by Schouwink. The patient had used at that time oral zinc sulphate for approximately 1.5 years. No changes in Kayser-Fleischer rings and ceruloplasmin levels were mentioned. Our findings suggest that oral zinc sulphate may not only prevent storage of copper in the tissues but may also contribute to the mobilization and excretion of deposits of copper.


Facial palsy as the initial symptom of a lymphoreticular malignancy. Case report. In this case report an adult patient is described who initially manifested a peripheral facial palsy mimicking 'Bell's palsy', which later on appeared to be the first manifestation of a lymphoreticular malignancy. Cranial nerve involvement appears to be very rare as a first symptom of malignant systemic disease. The importance of additional investigations including lumbar puncture and CSF cytology in cases of facial nerve palsy is stressed.


Effect of partial gastric devascularization on mucosal blood flow and acid secretion in cats. Partial devascularization of the stomach was performed in cats anesthetized with pentobarbital. The devascularization included all blood vessels along the greater curvature and two of the main branches of the left gastric artery. Gastric mucosal blood flow and cardiac output were determined by means of the microsphere distribution technique. The acid (H+) output from the 30th to the 60th min of pentagastrin stimulation was used as the 'acid response value'. Blood flow and acid secretion were determined before and approximately 2 h after devascularization or sham operation. A marked decrease in acid secretion and mucosal blood flow of the corpus/fundus occurred after devascularization. A high degree of correlation was found between the decrease in acid secretion and the decrease in mucosal blood flow caused by the devascularization. The regression coefficient of acid secretion on mucosal blood flow did not change significantly after devascularization.


Experimental acute pancreatitis: action of atropine and beta-haloethylamine furoate on muscarine-induced exocrine pancreatic secretion. Intraarterially administered muscarine induced predictable and reproducible experimental acute pancreatitis with a simultaneous increase in amylase levels in blood. Muscarine also caused a transient rise followed by a lowering of blood glucose levels. The stimulated amylase secretion was dose-response related. The guinea pigs survived 2--2 1/2 h after muscarine administration. Atropine (3 and 5 mg/kg), an antimuscarinic agent, injected intraperitoneally 2 h prior to muscarine administration, (a) inhibited muscarine-induced amylase secretion, and (b) marginally increased the survival time of guinea pigs, but could not sustain the animals for further experimentation. The high death rate of experimental animals prevented the use of this method as a model for investigation of experimental acute pancreatitis.


Early metabolic disturbances in the liver following unilateral hepatic or common bile duct obstruction in rabbits. The early metabolic disturbances in the liver following ligation of the left hepatic or the common duct were studied in the rabbit. In the rabbits subjected to ligation of the left hepatic bile duct, the phosphorylative activity of the liver mitochondria in the ligated lobe decreased to about 50% of controls 48 h after the ligation, while that in the nonligated lobe increased 2-fold. These rabbits continued to live. On the other hand, in the rabbits subjected to ligation of the common bile duct, the mitochondrial phosphorylative activity increased slightly at 3 h after the ligation but fell rapidly thereafter. The mitochondrial free NAD+ to NADH ratios and hepatic energy charge (ATP+1/2ADP/ATP+ADP+AMP) markedly decreased after the ligation. Most of these rabbits did not survive over 2 days. These results indicate that the metabolic disturbances in the jaundiced liver are due to an impairment of the oxidative phosphorylation. In addition, it is suggested that the maintenance of the energy balance by enhanced mitochondrial phosphorylative activity is readily impaired in the jaundiced liver.


Some functions of canine liver perfused ex vivo with bloodless fluorocarbon emulsion. 6 canine livers were perfused extracorporeally with bloodless fluorocarbon emulsion for 6 h. Perfusate was oxygenated in vitro with a mixture of oxygen and carbon dioxide. Biochemical studies were performed on the samples of perfusate taken from the arterial and venous lines of the perfusion circuit, and of the bile. Hepatic tissue was sampled prior to and following 6 h of perfusion for histological and biochemical studies. Changes, if any, found in the concentrations of various components of perfusate and bile, were considered due to the metabolic activity of the liver. Moderate changes of the liver architecture and a decrease of glycogen were found in hepatic tissues studied microscopically. Under the conditions of the present preliminary experiment, canine livers remained functional after 6 h of perfusion.


Effects of somatostatin on gastric mucosal histamine and serotonin content in restrained rats. The effects of continuous intravenous perfusion of somatostatin on the incidence of ulcers and the histamine and serotonin gastric mucosal content in rats subjected to 12 h of restraint were studied. 51 sets of Sprague-Dawley male rats (102 animals) were divided at random into two groups. Those in group I were immobilized and treated with an intravenous infusion of isotonic saline solution. Animals in group II were also immobilized and given intravenous cyclic somatostatin dissolved in isotonic saline solution; the initial dose was 25 microgram and a continuous infusion of 2.5 microgram/kg/h was administered thereafter. A decrease in the ulcer index was observed in the rats perfused with somatostatin (p less than 0.01) as well as a decrease in the histamine (p less than 0.001) and serotonin (p less than 0.01) gastric mucosal contents. These effects of somatostatin on restraint-induced stress ulcer in rats are probably to be explained by microcirculatory changes in the gastric mucosa determined by the vasoactive amines (histamine and serotonin). They constitute an important factor in the pathogenesis of these lesions.


Systolic time intervals in middle-aged men according to smoking habits and other risk factors of coronary heart disease. In the course of a prospective study on coronary heart disease (CHD) risk factors, systolic time intervals (STI) were measured at rest in 92 men aged 52--56 yr and free of any clinical symptoms of CHD. The ratio preejection period/left ventricular ejection period (PEP/LVEP) was significantly augmented in current smokers vs nonsmokers (0.406 +/- 0.075 vs 0.384 +/- 0.068, P less than 0.05). The regression of this parameter on cigarette consumption per day, taking into account heart rate, systolic blood pressure, body mass index, serum cholesterol and triglycerides, remains significant. An acute experiment, involving 50 smokers recruited similarly, showed no significant variation of the STI after the smoking of 2 cigarettes. We concluded that the augmentation of the PEP/LVEP ratio may represent a chronic effect of tobacco smoking on cardiac function.


'Bradycardia-tachycardia' syndrome 8 yr after correction of Fallot's tetralogy. A 13-yr-old boy presented with atrial flutter 8 yr after surgical correction of tetralogy of Fallot; antiarrhythmic therapy caused depression of the sinoatrial node, with syncope. Disordered sinoatrial function and intraventricular conduction were demonstrated by intracardiac electrography, and appear to have resulted from the operation. Sinoatrial disease may be responsible for supraventricular arrhythmias or syncope long after operative correction of Fallot's tetralogy and may be one of the explanations for the tendency of such patients to die suddenly.


Sinus node disease affecting both parents and both children. A family is described in which both parents and both of their children developed conducting system disease before the age of 40. They all had evidence of possible sinus node dysfunction. The father and son had an unexplained polyneuritis during childhood.


Coarctation of the aorta operated upon in infancy. Long-term follow-up. A long-term follow-up study was performed in 14 patients, operated for coarctation of the aorta in infancy. The median age at operation was 4 mth. Mean and median age at follow-up were 16 yr, and the follow-up time varied between 7 and 22 yr (mean and median 15 yr). At follow-up recoarctation was or had been present in 6 patients; there was no relation between the development of recoarctation and the age at operation. Concomitant aortic valve anomaly had been overlooked at the initial investigation in 3 patients. The systolic and diastolic blood pressure at follow-up was significantly elevated above the mean of normals; 2 of 8 patients without recoarctation clearly had diastolic hypertension. It seems as if even operation in infancy does not prevent hypertension later in childhood or adolescence in some of the patients, and it hardly seems justified to perform elective operations for coarctation of the aorta in infancy with the appreciable risk of recoarctation, until more long-term follow-up results are known. The patients should not be lost to follow-up.


Canrenoate--a spironolactone metabolite. Acute cardiac effects in digitalized patients. Canrenoate (Soldactone)--a spironolactone metabolite for intravenous use--has recently been suggested as a specific antidote in patients suffering from digitalis intoxication. Theoretically, this would imply that canrenoate could counteract the effect of digoxin. The influence of canrenoate on inotropy in 12 digitalized patients was investigated by measurement of sytolic time intervals. Canrenoate improved the left ventricular contractility, since both preejection period (PEP) and the ratio between PEP and left ventricular ejection time (PEP/LVET) decreased significantly. The maximal changes in controlled atrial fibrillation, canrenoate had no effect on ventricular rate, whereas atropine increased the ventricular rate significantly.


An experimental model for angina pectoris using krypton-81m. The dynamic balance between myocardial perfusion and metabolism. 16 anesthetized and open chest dogs were studied. Regional myocardial perfusion was assessed using a constant infusion of krypton-81m (half-life 13 sec) into the aortic sinuses and a gamma camera linked to a digital computer. The epicardial electrocardiogram was recorded and the plasma activity of creatine kinase was measured in serial blood samples from the aorta and a local coronary vein draining the area of myocardium supplied by the left anterior descending coronary artery (LAD). These parameters were observed throughout the whole period of a 5-h experiment. Two reversible snares were positioned on the middle portion of this artery. A critical narrowing of this vessel was produced and a peripheral venous infusion of isoproterenol (causing a 5--10% increase in heart rate and a 10--15% fall in blood pressure) was used to increase myocardial oxygen demand. During infusion there was both a relative and absolute fall in regional myocardial perfusion together with evidence of myocardial ischemia in the epicardial electrocardiogram. Provided the infusion was discontinued within 30 min (8 dogs) myocardial perfusion and the epicardial electrocardiogram returned to normal during a 5-h recovery period. In addition there was no efflux of creatine kinase activity from the ischemic area. When infusion was continued for 1 h (4 dogs) permanent alterations in myocardial perfusion and the epicardial electrocardiogram occurred and there was increased creatine kinase activity released from the area of myocardium by the narrowed vessel. Infusion for 40 min in 4 dogs produced permanent alterations in the parameters measured in 2 and complete recovery in the remaining 2. A further 4 dogs were studied in the same way but without a snare on the coronary artery. Isoproterenol given for 1 h produced no effects on any of the parameters either during or after infusion.


Effect of respiration on echocardiographic ventricular dimensions and relationship to the second heart sound. The effects of normal respiration on echocardiographic right and left ventricular dimensions were studied in 8 normal subjects (group 1) and in 10 patients with ventricular volume overload due to anaemia (group 2). Right ventricular internal diameter (RVID) increased on inspiration in both groups of patients (group 1, 0.3 +/- 0.1 cm; group 2, 0.6 +/- 0.4 cm), while left ventricular internal diameter (LVID) decreased (group 1, 0.3 +/- 0.2 cm; group 2, 0.4 +/- 0.3 cm). The changes in echocardiographic diameter were related to the duration of electromechanical systole and to the changes in the splitting of the second heart sound; there was a linear relationship between RVID and Q--P2 (P less than 0.001) and A2--P2 (P less than 0.001). Echocardiographic measurements of cardiac chamber size should take into account the changes which occur during respiration.


Effect of nifedipine on plasma renin, aldosterone and catecholamines in arterial hypertension. Acute sublingual administration of nifedipine 10--20 mg to 13 hypertensive patients caused a rapid decrease in blood pressure (BP) and a concomitant increase in heart rate (HR), plasma noradrenaline (NA) and plasma renin activity (PRA); there was no significant change in plasma adrenaline (A) or aldosterone (ALDO). Basal PRA was the major determinant of the rise in PRA, as a close correlation was present between the basal value and the increase caused by nifedipine (r = 0.92), p less than 0.001). The rise in PRA was also correlated with the plasma concentration of nifedipine after 60 min (r = 0.80, p less than 0.01), but it was not correlated with the decrease in BP, the rise in HR or the increase in NA. Nifedipine 30--60 mg daily for 6 weeks caused a reduction in mean BP from 133 to 113 mmHg (p less than 0.001). Body weight and serum potassium decreased but no consistent change was noted in NA, PRA, ALDO or 24 h-excretion of catecholamines. A significant correlation was present between the change in NA and that in PRA (r = 0.74, p less than 0.01). The alterations in the various parameters in the acute and chronic studies were not correlated. The findings indicate that different regulatory mechanisms are activated during acute and chronic administration of nifedipine. It is suggested that an initial rise in sympathetic activity gradually decreases during prolonged therapy, but it still remains a determinant of PRA.


Intracardiac electrophysiological effects of lorcainide in man. The electrophysiological effects of lorcainide 1.25 or 2.5 mg/kg given iv over 2 or 4 min, were studied in 21 patients with normal and diseased impulse formation and conduction, by means of intracardiac recording and stimulation. Sinus rate and the effective atrial refractory period rose following both doses of lorcainide. The corrected sinus node recovery time rose only after lorcainide 2.5 mg/kg and then most markedly in patients with sinus node dysfunction. The P-A interval remained unchanged following the drug. The A-H interval during sinus rhythm, and the pooled A-H intervals during atrial pacing, increased slightly, and the functional and effective A-V nodal refactory period changed variably. Wenckebach periods above the bundle of His occurred at lower atrial pacing rates following both doses of lorcainide in 7 patients, at the same atrial pacing rate in 9 and at higher rates in 3. H-V intervals, pooled H-V intervals and QRS-width lengthened in all patients, most markedly in cases with a conduction delay below the His bundle, who had received lorcainide 2.5 mg/kg. Thus, lorcainide shares some electrophysiological properties with procainamide and aprindine. Higher doses should be used with caution in patients with pre-existing conduction delay below the bundle His.


Why are psychotropic drugs prescribed to out-patients? A methodological study. The prescription of psychotropic drugs at a multidoctor district health centre in northern Sweden in 1973, was analysed by means of problem-oriented medical records. Of the 22,000 inhabitants of the district 10,700 consulted the health centre. Psychotropic drugs were prescribed for 11.3% of the patients, corresponding to 5% of the inhabitants of the area. Sixty per cent of the patients received one psychotropic prescription and 90% not more than three. Two-thirds of prescriptions were for women. Hypnotics, sedatives and minor tranquillisers constituted 64% of all prescriptions, major tranquillisers 24% and antidepressants 12%. One fifth of the patients obtained drugs belonging to more than one of the major psychotropic groups during the year. Insomnia, psychoneurosis and depression made up two-thrids of the indications for psychotropic drug therapy. More than thirty different psychotropic drugs were prescribed for the two major indications. There was considerable variation in how the different doctors prescribed drugs for the same indication. Fifty-nine different drug products were prescribed, of which the commonest five constituted more than half of the total number. Individual doctors used from 22 to 38 different psychotropic drugs.


Blood level of cimetidine in relation to age. The blood level versus time curve for unchanged cimetidine after a 200 mg oral dose has been determined in 20 apparently healthy subjects, ranging from 22 to 84 years of age. A significant relationship between the area under the curve (AUC) and age was found (r = 0.81; P less than 0.001). The peak concentrations of cimetidine were significantly inversely related to body weight (r = -0.71; P less than 0.001). The age-related increase in bioavailability of oral cimetidine, as measured by AUC, was probably due to decreased total clearance of the drug, which resulted from the opposed changes (by themselves not significant) of distribution volume towards a decrease and of half-life towards an increase with age. Reduction in the standard oral dose of cimetidine by one third to one half should be feasible in the elderly without loss of efficacy, and it may be advisable in order to obviate extreme individual responses that may occur in this population.


Influence of food on the absorption of phenytoin in man. The influence of food intake on the absorption of phenytoin was examined in eight healthy volunteers, by study of single-dose kinetics following ingestion of phenytoin 300 mg either with a standardized breakfast or on an empty stomach. Blood samples were collected at regular intervals from 0 to 48 h, and serum concentrations of unmetabolized phenytoin were determined by gas chromatography. Serum concentrations of the major metabolite of phenytoin, 4-hydroxyphenytoin, were measured by mass fragmentography. Concurrent intake of food and phenytoin appeared to accelerate absorption of the drug from the formulation used, and the peak concentrations were significantly higher (mean increase 40%) in the postprandial than in the preprandial state. As reflected by the AUC (area under the curve), the amount of drug absorbed was increased during postprandial conditions, although the difference only reached borderline significance. It is suggested that phenytoin should always be taken in a defined relation to meals.


Pharmacokinetics of bromocriptine during continuous oral treatment of Parkinson's disease. The plasma kinetics of bromocriptine (BCT), a long-acting dopamine agonist, was studied in twelve patients with Parkinson's disease, using a newly developed gas chromatographic method of analysis. Each patient received BCT for at least three weeks in a constant but different dose regimen. Concomitant treatment with 1-DOPA was not allowed. During a 6-day hospitalization period, a blood sample was taken immediately before the afternoon dose at 14.00 h (Cmin) to determine the steady-state level. On the 6th day blood samples were collected every hour during two 8 h dose intervals. The results showed a significant correlation between the mean values of the AUC and the Cmin. First order elimination kinetics appeared to be followed by BCT, at least for the plasma concentrations commonly found. Considerable inter-individual variation was demonstrated both for the dose/plasma concentration ratio and for calculated plasma clearances. No serious side-effects were observed during the investigation.


Excretion of hydroflumethiazide in bile and urine of man. Biliary and urinary excretion of hydroflumethiazide (HFT) and its metabolite, 2,4-disulfamyl-5-trifluoromethylaniline (DTA), was investigated in 5 otherwise healthy patients with a T-drain in the common bile duct after cholecystectomy and choledochotomy. After a single oral dose of HFT 302--453 micromoles, a mean of 0.051% (range 0.028--0.075) of the dose was excreted in bile and 34.9% (range 23.7--45.4) in urine during the first 6 hours after administration. Biliary excretion appeared to be of minor importance in the elimination of HFT by man. DTA could not be detected in bile.


An opposing role for the adrenals in the hypotensive effects of propranolol in the spontaneously hypertensive rat. d,l-Propranolol (1 and 5 mg/kg s.c.) did not cause a fall in blood pressure and induced only a limited decrease in heart rate in conscious spontaneously) hypertensive rats (SHR). In contrast, after bilateral adrenalectomy, d,l-propranolol induced a rapid and profound decrease in blood pressure and heart rate. Decreases in heart rate and blood pressure in the individual animals were not correlated. The effects were mainly caused by l-propranolol but an additional effect of d-propranolol cannot be excluded. The decrease in blood pressure was not observed after removal of the adrenal medulla. Heart rate decreased only slightly in these animals. After treatment of adrenalectomized SHR with corticosterone (1 mg/kg b.w./h) the decrease in blood pressure due to d,l-propranolol was completely abolished. The fall in heart rate was diminished. Central injection of d,l-propranolol into the lateral brain ventricle of adrenalectomized SHR caused cardiovascular changes which were less pronounced than those following peripheral injection of comparable doses. The inhibitory effects of d,l-propranolol also occurred in adrenalectomized normotensive Wistar Kyoto rats. However, no significant changes in blood pressure and only a limited fall in heart rate were observed in adrenalectomized normotensive and renal hypertensive Wistar rats. It is concluded that the presence of the adrenal cortex, but not of the adrenal medulla prevents acute hypotension and bradycardia after propranolol in the conscious SHR.


Inhibition of gastric acid secretion by a glycoprotein isolated from human urine (human urinary gastric inhibitor). The gastric antisecretory activity of an inhibitor newly isolated from human urine (Human Urinary Gastric Inhibitor or HUGI) has been studied. HUGI was given intravenously and its activity determined in the following test systems: gastric secretion in the rat with pyloric ligation; gastric secretion in the dog with a Heidenhain pouch stimulated with pentagastrin, histamine and a protein meal; acid secretion by the isolated gastric mucosa of the rat; gastrointestinal motility; bile flow and gall-bladder tone and arterial and venous blood pressure and heart rate. HUGI was found to have marked activity only in the pyloric-ligated rats and in the dogs with Heidenhain pouches stimulated by a protein meal. Particularly in the dog, HUGI (0.1 to 6.4 micrograms/kg, i.v) markedly inhibited gastric secretion, dose-dependently and without changing the plasma gastrin concentration. Negative results were obtained in the other tests, but these results serve to demonstrate that HUGI is an inhibitor well-differentiated from other glycoproteins or peptides with gastric antisecretory activity, such as urogastrone and GIP. The results obtained to date are not sufficient to allow the mechanism of action of HUGI to be defined.


3,4-Dihydroxyphenylacetic acid and homovanillic acid in rat plasma: possible indicators of central dopaminergic activity. The concentrations of dopamine (DA) metabolites (free and conjugated) was measured in plasma and brain regions of rats by the mass spectrometric method of selected ion monitoring. Experimental treatments which altered the function of central dopamine neurons also induced concomitant changes in plasma 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA). Stimulation of the nigrostriatal pathway increased plasma DOPAC and HVA whereas lesion of the pathway decreased plasma metabolites. Several drug treatments induced parallel changes in brain and plasma concentrations of DA metabolites. It is suggested that changes in the concentration of DOPAC and HVA in rat brain are reflected by parallel changes in plasma. No conjugated forms of DOPAC and HVA were found in plasma and brain tissue of vervet monkeys (Cercopithecus aethiops).


Mobilization of stored calcium for phasic contraction induced by norepinephrine in rabbit aorta. In Ca-free solution norepinephrine (NE) produced only a phasic contraction in the media-intima layer of rabbit aorta. The second application of NE was almost ineffective. Incubation of the muscle with Ca for a short period (Ca loading) restored the ability to produce a phasic contraction in Ca-free solution. Various ions and compounds were applied to the muscle prior to the Ca loading or prior to NE application and it was found that these treatments variously affected the NE-induced phasic contraction. The data suggest that the NE-induced phasic contraction in Ca-free solution results from Ca release from cellular sites and that a hyperbolic relationship exists between the amount of Ca at these sites and the magnitude of the contraction. These sites take up extracellular Ca by a process sensitive to La3+ but not to D600; the Ca influx stimulated by high K solution, which is sensitive to D600 and also contributes to refilling of these sites; NE releases this stored Ca by a process sensitive to procaine, caffeine and theophylline and, in this manner, elicits a phasic contraction.


Analysis of the effects of bromocriptine on blood pressure and sympathetic nerve function. Bromocriptine, a dopamine receptor agonist, lowered arterial blood pressure and caused significant impairment of the cardioacceleration observed during the stimulation of the right postganglionic cardiac sympathetic nerve fibers in pentobarbital-anethetized dogs. Prior administration of pimozide antagonized the hypotensive and the inhibitory action of bromocriptine on cardiac sympathetic function. These results suggest that while presynaptic dopamine receptors seem to be involved in mediating the inhibitory action of bromocriptine on cardiac sympathetic function, the role of presynaptic as well as dopamine receptors located at other sites in mediating the hypotensive action of bromocriptine needs to be further investigated.


Vasodilator response to histamine: dependence upon the site of administration. Histamine causes vasodilation in part by interacting with histamine H2 receptors. The present study was undertaken to evaluate the difference in vasodilator sensitivity of H2 receptors on the inside compared to the outside of the vessel. In order to induce tone, norepinephrine was administered to an in vitro preparation of rabbit ear artery treated with mepyramine to block H1 receptors. Histamine was then either selectively perfused through the artery or added to the outside of the vessel via the organ bath. The outside of the artery was found to be twice as sensitive to the vasodilator effect of histamine as the inside. These data provide the first evidence for greater extraluminal sensitivity of vascular smooth muscle and suggest that the response to histamine will depend in part on the route the amine takes to reach receptors, e.g., from blood-borne sources or from extraluminal stores.


Sulpiride: a weak antagonist of norepinephrine and 5-hydroxytryptamine. Antagonism of norepinephrine (NE) and 5-hydroxytryptamine (5-HT) by sulpiride was studied in vitro on rabbit aortic strips. Concentrations of sulpiride ranging from 3 X 10(-5) to 3 X 10(-3) M caused a progressive shift of the dose response curves of both NE and 5-HT to the right without inhibiting the responses to KCl. pA2 values of sulpiride calculated from Schild plots against NE and 5-HT were 4.6 and 4.4, respectively. Thus, sulpiride, which has been previously shown to be the most potent antagonist of the dopamine vascular receptors, is a very weak alpha-adrenergic or 5-HT antagonist.


Enhancement of in vitro binding and some of the pharmacological properties of diazepam by a novel anthelmintic agent, Avermectin B1a. A novel macrocyclic lactone disaccharide anthelmintic agent, Avermectin B1a (AVM) has been found to cause a concentration-dependent increase in the in vitro binding of 3H-diazepam to rat and mouse brain membranes. The increase in binding is manifested as both an increase in the affinity and number of bindings sites for 3H-diazepam. Preliminary in vivo studies demonstrate that AVM can also enhance some of the pharmacological actions of diazepam.


Cooperative interactions at [3H]kainic acid binding sites in rat and human cerebellum. Inhibition of the specific binding of [3H]kainic acid was studied in membranes isolated from rat and human cerebellum; the sequence of potencies in both species were: kainic acid greater than L-glutamic acid greater than dihydrokainic acid greater than D-glutamic acid. Whereas the Hill coefficient for unlabelled Kainate was 1.0, dihydrokainic acid and D- and L-glutamic acids exhibited negative cooperativity with Hill coefficients of near 0.5. This allosteric interaction of glutamic acid at the kainic acid recognition site suggests a biochemical correlate for the synergistic effects of these compounds in vivo.


Vasopressin analogs: sedative properties and passive avoidance behavior in rats. The effects of several types of vasopressin analogs that are considered to be resistant to some of the physiologically significant enzymatic systems were investigated utilizing rats trained in a passive avoidance task. Enhancement of avoidance latencies was observed 2, 7 and 13 days after the single learning trial when deamino-carbavasopressins, triglycyl-8-lysine-vasopressin or its des-glycinamide derivative, and deamino-D-arginine-vasopressin were given shortly after the learning trial in the dose of 1 microgram s.c. (8-L-Arginine)deamino-6-carba-vasopressin and (8-L-ornithine)deamino-6-carba-vasopressin were also active in the dose of 0.1 microgram. Lysine vasopressin and its des-glycinamide derivative failed to enhance avoidance latencies in part of the experiments if doses of 0.3--3 micrograms were administered and 7 or 13 day intervals were used between the learning and the test trials. Enhancement of avoidance latencies was also observed, if some of the peptides were injected 20 min but not 120 or 180 min before the test trial. Marked depression of exploratory behavior of rats in an open field was found after s.c. injections of low doses (1--3 micrograms kg-1) of deamino-carba-vasopressins. Higher doses (10--30 micrograms kg-1) induced sleep-like immobility not accompanied by ataxia or catalepsy.


Effects of chlorpromazine and some of its metabolites on the EEG and on dopamine metabolism of the isolated perfused rat brain. The study concerned the effects of chlorpromazine (CPZ), monodesmethyl-chlorpromazine (NOR1-CPZ), didesmethyl-chlorpromazine (NOR2-CPZ), and chlorpromazine-N-oxide (CPZ-NO) on the EEG and on dopamine metabolism of the isolated perfused rat brain. Isolated brains were perfused with 100 ml of a perfusion medium containing 30% bovine red cells (v/v), 2 g bovine serum albumin, 14 mM glucose as well as one of the agents in a concentration of 10 micrometers. The main dopamine metabolite homovanillic acid (HVA) was measured fluorimetrically in the striatum of the isolated brain. The EEG was recorded by two symmetrical bipolar leads from the parietal regions at various times during the 30 min perfusion period and was stored on magnetic tape. The recordings were evaluated visually and quantitatively by automatic analysis. CPZ-NO was found to be the most active agent both in changing the EEG and in elevating the HVA level in the striatum. The mean EEG amplitude and the slow wave activity increased significantly. The increase of the HVA level in the striatum was correlated with the increase of delta waves as well as excess of kurtosis and skewness calculated from the amplitude histography data. The desmethylated metabolites caused only moderate central effects.


Mechanism of the depressor response to dopamine in the rat treated with phenoxybenzamine. Alterations of hypotensive responses to dopamine by antagonists were characterized in alpha-blocked, anaesthetized rats. Responses were not affected by d-propranolol (0.1 mg/kg) whereas d,1-propranolol (0.1 mg/kg) or haloperidol (1.0 mg/kg) attenuated them; higher doses of inhibitors (1.0 mg/kg; 5.0 mg/kg, respectively) failed to produce a higher inhibition, but combinations of low doses abolished the depressor responses. In adrenalectomized rats, hypotensive responses decreased; haloperidol always attenuated the responses while d,1-propranolol became ineffective. Dopamine produced an enhancement of plasma adrenaline and noradrenaline levels, which was decreased by d,1-propranolol and increased by haloperidol. The data suggest that in rats the depressor component of dopamine is due to activation of both dopaminergic and beta-adrenoceptors. The beta component appears to be due to the release of adrenaline. The results also support the concept of the existence in sympathetic nerve endings and adrenal glands of stimulatory beta-adrenergic and inhibitory dopaminergic prejunctional receptors.


Comparison of binding of [3H]-methionine-enkephalin, [3H]-naltrexone and [3H]-dihydromorphine in the mouse vas deferens and the myenteric plexus and brain of the ginea pig. [3H]-Methionine-enkephalin, [3H]-naltrexone and [3H]-dihydromorphine are specifically bound in homogenates of not only the brain and myenteric plexus of the guinea pig but also the vas deferens of the mouse. Brain has a ratio of methionine-enkephalin to dihydromorphine binding in favour of methionine-enkephalin binding and the myenteric plexus a ratio in favour of dihydromorphine binding, with the mouse vas deferens being intermediate.


The effects of intra-arterial bradykinin, histamine, acetylcholine and prostaglandin E1 on nociceptive and non-nociceptive dorsal horn neurones of the cat. The effects of peripherally administered algesic agents were investigated on the firing of cat dorsal horn interneurones classified as nociceptive or non-nociceptive according to the peripheral stimuli that excited them. A small amount of bradykinin injected into the blood supplying the receptive fields of cells was a potent specific stimulus causing activation of nociceptive cells and slowly conducting nerve fibres. Larger amounts of bradykinin and large amounts of histamine, 5-hydroxytryptamine and acetylcholine activated both nociceptive and non-nociceptive cells. Prostaglandin E1 enhanced the effects of bradykinin and histamine on nociceptive cells. Prostaglandin E1 also increased the response of these cells to the application of noxious heat whilst aspirin reduced this response. These results support a chemosensitive theory of nociceptor activation and show bradykinin to be the most potent and specific of the suggested endogenous algesic agents in causing activation of CNS nociceptive pathways.


Neurotensin contracts the guinea-pig longitudinal ileal smooth muscle by inducing acetylcholine release. Neurotensin contracted the isolated longitudinal smooth muscle strip of the guinea-pig ileum, but was only 22% as effective as histamine. Neostigmine (0.1 micrometer) increased the effectiveness of neurotensin 4-fold. Atropine (0.1 micrometer) completely inhibited the contracting effect of neurotensin measured in the presence of neostigmine, whereas hexamethonium (0.1 micrometer) was without effect. It is concluded that in the longitudinal smooth muscle of the guinea-pig ileum, the contracting effect of neurotensin is mediated by the release of acetylcholine from nerve endings in response to a postganglionic stimulation of cholinergic nerve fibers.


Effects of disopyramide on SA nodal pacemaker activity and contractility in the isolated blood-perfused atrium of the dog. The isolated blood-perfused preparations of canine atrium were suspended in a bath and perfused with arterial blood led from the carotid artery of the heparinized donor dog. Disopyramide caused dose-related negative chronotropic and inotropic effects in a dose range of 30-1000 microgram when injected directly into the cannulated sinus node artery of the isolated atrium. The order of potencies for inducing the negative chronotropic effect in isolated atrium preparations was verapamil greater than propranolol greater than lidocaine = quinidine greater than phenytoin greater than or equal to disopyramide greater than procainamide. On the other hand, the order of potencies for inducing the negative inotropic effect was verapamil = propranolol greater than lidocaine greater than or equal to phenytoin greater than disopyramide greater than procainamide greater than or equal to quinidine. When disopyramide (1 mg/kg or 3 mg/kg) was administered i.v. into the jugular vein of the donor dog, the systemic blood pressure of the donor dog was markedly decreased. However, the tension developed and sinus rate of the isolated atrium were only slightly decreased. Disopyramide produced greater suppression at higher frequencies and slightly depressed the calcium chloride-induced positive inotropic effects.


Central mediation of the antihypertensive effect of pargyline in spontaneously hypertensive rats. The monoamine oxidase (MAO) inhibitor pargyline induced a moderate (about 20 mm Hg) but persistent (48 h) decrease of systolic blood pressure in unanesthetized adult spontaneously hypertensive rats (SHR) but not in normotensive rats. The fall of blood pressure correlated with the blockade of norepinephrine (NE) deamination by brain homogenates. After an intracerebroventricular (icv) injection of 6-hydroxydopamine, which lowered brain NE content by about 70%, pargyline was unable to diminish arterial pressure. Blockade of central alpha-adrenoceptors by treatment with phentolamine (100 microgram icv) could either prevent or reverse the fall of blood pressure in SHR induced by pargylline. Moreover, a low dose of pargyline injected directly into the brain lowered arterial pressure. We conclude that the hypotensive action of pargylline in SHR appears to be the consequence of NE accumulating at an inhibitory alpha-adrenoceptor in brain.


Physiological function of descending noradrenaline projections to the spinal cord: role in post-decapitation convulsions. Destruction of the descending noradrenergic innervation to the spinal cord, but not that to the cerebellum or the forebrain, by the use of intracerebral injection of 6-hydroxydopamine completely prevented the occurrence of the usual itation convulsion. Depletion of brain noradrenaline by synthesis inhibition with DDC, FLA 57 or FLA 63 g reduced the duration of the post-decapitation convulsion. Blockade of alpha-noradrenergic receptors by phentolamine or phenoxybenzamine, but not of beta-receptors by propranolol, also reduced the duration of the convulsion. The presynaptic alpha-agonist, clonidine, at either 1 mg/kg or 0.05 mg/kg also reduced the magnitude of the convulsion but either blockade of dopamine receptors with pimozide or destruction of the ascending dopamine systems by 6-hydroxydopamine was without effect. It is concluded that dopamine systems are not involved in post-decapitation convulsions and that the noradrenergic involvement is by the descending spinal projections acting on a post-synaptic alpha-receptor in the spinal cord, but also modulated by presynaptic alpha-receptors possibly on the locus coeruleus perikarya.


GABA modulates the release of dopamine and acetylcholine from rat caudate nucleus slices. The effects of GABA on depolarization-induced (26 mM K+) release of radiolabeled dopamine (DA) and acetylcholine (ACh) from slices of rat caudate nucleus were examined with a superfusion method. GABA, in concentrations of 10(-5)--10(-3) M, dose-dependently enhanced the release of DA, either accumulated by high-affinity uptake or synthesized from 14-C-tyrosine. In contrast, the release of ACh was reduced by GABA. This reduction appeared to be caused by the increase in DA-release. These effects of GABA decreased from the caudal to rostral part within the caudate nucleus, an order which parallels the distribution of endogenous GABA and glutamic acid decarboxylase. However, GABA had little, if any, effect in the nucleus accumbens. Since it was difficult to antagonize the effects of GABA on DA and ACh release with bicuculline or picrotoxin, it remains uncertain whether these effects were mediated via GABA receptors. In view of the high endogenous GABA level in the caudate nucleus it is concluded that GABA may be one of the local factors involved in the control of the amount of transmitter that will be released from dopaminergic varicosities upon depolarization.


Analgesia produced by microinjection of baclofen and morphine at brain stem sites. Microinjection of either baclofen (1.5 microgram) or morphine (2.5 microgram), in equimolar doses (14 mM), at sites located in the caudal periaqueductal gray (PAG) resulted in a delay in tail flick latency (analgesia). The relative analgesic potency of baclofen among caudal PAG sites, however, did not correlate with that of morphine. Application of either drug into the caudal aspect of the cerebral aqueduct also produced analgesia, but neither agent caused analgesia when applied at PAG sites rostral to the interaural line. Baclofen also produced analgesia when microinjected in the lower brain stem at sites lateral to the midline in or near the nucleus gigantocellularis, but did not produce analgesia when applied on the midline at sites located within or near the raphe magnus. Conversely, morphine produced analgesia when applied locally at midline sites but not at sites located lateral to the midline. These data suggest that the analgesia produced by systemic administration of baclofen and morphine involves activation of different neuronal substrates.


Action of indapamide on excitation-contraction coupling in vascular smooth muscle. The effects of indapamide on electrophysiological and mechanical parameters of longitudinal smooth muscle strips isolated from mammalian portal vein were studied by means of a double sucrose gap method associated with a photoelectric device for recording contraction. Indapamide (10(-4) M) reduced both the amplitude of the action potential and the contraction. The calcium inward current decreased and consequently the phasic contraction was also reduced. The potassium outward current was diminished while the tonic contraction was not modified significantly. The depressant nature of the indapamide response could counterbalance the stimulating action of angiotensin II but not that of noradrenaline. The results suggest  that indapamide acts primarily on the plasma membrane of vascular smooth muscle by reducing the transmembrane calcium current, although a secondary decrease in the intracellular bound calcium could not be completely excluded.


Effects of alkyl analogs of histamine and metiamide on gastric acid secretion. Histamine and metiamide analogs with alkyl, methyl, ethyl or isopropyl groups on the imidazole ring in position 5, not previously tested on an in vivo preparation, were studied for their effects on gastric acid secretion in cats with gastric fistulas. Our findings confirmed that the methyl group appears to be optimal for an effect on gastric acid secretion as regards both agonism and antagonism. Ethyl substitution apparently did not change the pharmacological activity of histamine but modified the inhibitory effect of metiamide from competitive to non-competitive kinetics. Isopropyl analogs were inactive on gastric secretion under our experimental conditions.


Genetic variability within and between outbred Wistar strains of rats. The allelic frequencies at 6 isozyme loci (Es-1, Es-2, Es-3, Es-4, Es-Si and Amy-1) were examined in 4 outbred Wistar strains and additionally 2 SD strains as controls. From the allelic frequencies, the averages of the heterozygosities in individual strains and the geometric genetic distances between every pair of all possible strain combinations were calculated. The averages of the heterozygosities in 2 SD strains were both intermediate (around 0.2) and the genetic distance between them was rather short. But among the Wistar strains, the averages of the heterozygosities were highly variable and the genetic relationships among them were very variable in their genetic distances. From these results, it was suggested that the outbred Wistar strains were different each other in their genetic constitutions and that no suggestion was obtained to descriminate genetically the Wistar strains from the SD strains.


[Comparison of different types automatic water-supply system for mouse rearing (author's transl)]. Rearing and breeding scores were compared between groups of mice (JCL : ICR and ddN strains) raised with two different types of automatic water-supply systems; the Japanese type and the American type, using manual water-supply system as control. The mice raised with the manual water-supply system were superior in body weight gain as compared to those with two automatic water-supply systems. As to the survival rate, however, the m;  anual water-supply system and the Japanese type gave better results than the American type. As to weanling rate in the breeding test, the manual water-supply system gave somewhat better result than either of the two automatic types. Accidental water leaks, which are serious problems of automatic systems, occurred frequently only when the American type was used. Only one defect of the Japanese type revealed was that it was unfavorable for mice with smaller size (e.g., young ddN mice), resulting in lower body weight gain as well as lower breeding scores.


[The incidences of spontaneous fetal anomalies in Japanese white rabbits (author's transl)]. The incidences of spontaneous fetal anomalies in coccidium-free Japanese white rabbits bred and kept in our laboratories are reported. Live fetuses were removed from 87 (95.6%) out of 91 maternal animals examined, totaling 538 (295 males and 243 females). of them, 9 fetuses (1.7%) showed external and visceral abnormalities such as hydrocephalia (5 cases), short tail (2), anophthalmia (1) and talipomanus flexa (1). Skeletal abnormalities were found in 7 (1.7%) of 424 fetues examined.


[A survey of Brucella canis infection in dogs sheltered in Tohoku University School of Medicine (author's transl)]. A total number of 1549 dogs from Miyagi Prefecture and the north coast area of Fukushima Prfecture were surveyed during a year from December 1976 to November 1977 for Brucella canis (B. canis). 173 of 1549 dogs (11.2%) were sero-positive to B. canis, and B. canis was isolated from 55 of 148 dogs examined, giving more than 3.6% for total 1549 dogs. The positivity were higher than that have been reported from other areas. The results of the morphological and biochemical studies for isolates were similar to the reference strain RM6/66, and they were identified as B. canis. No significant difference in sero-positive rate was found between male and female, but isolation rate was higher in female than in male. B. canis was isolated most frequently from the uterus and the spleen of both sexes. The importance of urine and milk as a source of infection was discussed as well as the role of infected focus at non gravid uterus for abortion and fetal infection. A case supposed to be a vertical infection was described also.


[Sturctural outline of spontaneous motor activity monitor using infrared photoelectric switch for small animals (author's transl)]. A simple monitor was developed for measuring the spontaneous motor activity (SMA) of small laboratory animals. The monitor was designed to measure animal ambulation along a horizontal axis by means of two infrared photo-beams. Using the monitor, male mice 7 to weeks of age at the commencement of this experiemtn showed high SMA values for the first 15 min and low values for the last 15 min of the experiment.


Quantitative development of adherent cell colonies in bone marrow cell culture in vitro. Quantification of the formation of adherent cell colonies in bone marrow cell culture was attempted. By secondary transfer of the bone marrow cells as a single cell suspension after 4 days' culture of fine marrow fragments, a linear relationship was obtained between the number of adherent cell colonies developing and the number of cells secondarily inoculated into the culture bottle. This suggests that 4 days' culture of the bone marrow cells with close intercellular interactions is sufficient for the 'conditioning' of the cells to develop adherent cell colonies. Activity of such colonies to support haemopoietic stem cell proliferation was also shown.


The role of hemin in the regulation of heme synthesis by fetal mouse liver erythroblasts in culture. The regulatory role of exogenous hemin on the heme synthetic pathway was studied in fetal mouse liver erythroblasts in culture. Hemin added to culture medium of 13th day embryo liver cells inhibited, dose dependently, the incorporation of the porphyrin precursors, 59Fe, 14C-2-glycine and 14C-5-aminolevulinic acid (ALA) by 85%, 70% and 45%, respectively. This suggests a multiple effect of hemin on the porphyrin biosynthetic enzymes. Exogenous ALA competed with 14 C-2-glycine as a porphyrin precursor, but the rate of heme synthesis, measured by 59Fe incorporation, remained unaltered. Protoporphyrin mimicked the hemin effect on the inhibition of glycine incorporation into heme, but reduced iron incorporation by only 20%. Erythroblasts, with an inhibited porphyrin biosynthesis, utilized exogenous 59Fe-hemin for hemoglobin assembly and maintained an undecreased level of hemoglobin synthesis. The results indicate that hemin inhibits the porphyrin biosynthesis in fetal mouse liver erythroblasts mainly at the iron incorporation stage.


The role of fibroblastoid cells and macrophages from mouse bone marrow in the in vitro growth promotion of haemopoietic tumour cells. Adherent cells from mouse bone marrow have been shown to promote the in vitro growth of the AVRij-1 tumour cell line. The experiments presented here suggest that the adherent cells involved in this phenomenon are the progeny of bone marrow derived fibroblastoid colony forming units. The latter were characterized by means of cell density distribution analysis. They had a broad distribution pattern and an average peak cell density of 1.069 g.cm-3. The growth promotion activity exerted by adherent cell layers from the various density fractions on the AVRij-1 tumour cell line coincided with the distribution of fibroblastoid colony forming units. On the other hand, the presence of macrophages in the adherent layers seemed to be non-essential for the in vitro promotion of growth of the AVRij-1 cell line.


Erythropoiesis in vitro. III. The role of potassium ions in erythroid colony formation. The role of potassium as an essential promotor of erythroid progenitor growth (BFU-e & CFU-e) from normal murine hematopoietic tissues was studied. Dialyzed fetal calf serum, over a wide range of concentrations, was shown to reduce the numbers of BFU-e and CFU-e that could be cultured from normal murine bone marrow. A dose-dependent addition of 1 M KC1 restored erythroid progenitor colony growth to the levels generally seen when normal, non-dialyzed fetal calf serum was used. Furthermore, when [K] was increased in some human urinary and sheep plasma erythropoietin preparations, the number of erythroid progenitor cells cultured also increased. This influence is crucial to the differentiation of committed stem cells into the erythroid pathway and must therefore be considered in the development of serum-free growth media.


Hemopoiesis of the cultured whole mouse embryo. In vitro cultured murine embryos provide a valuable methodology for studying the ontogeny of hemopoiesis in mammalian embryos. Under improved conditions, 49% of mouse embryos cultured from the blastocyst stage reached the early somite stage. The development of the embryo was arrested at different stages by omitting human cord serum from the culture medium at specific times. These observations suggest the appearance and differentiation of the first hemopoietic cells in the mouse yolk sac to be independent of a normal embryo. In the yolk sac of all cultured mouse embryos, hemopoietic cell differentiated into a major cell type, erythroblasts, and minor cell types such as monocytes, macrophages, and granulocytic cells. A small number of hemopoietic cells remained undifferentiated.


Lithium carbonate increases marrow granulocyte-committed colony-forming units and peripheral blood granulocytes in a canine model. Psychiatric patients given lithium carbonate (LC) commonly develop granulocytosis of the peripheral blood. This is an important observation with several potential applications in clinical hematology and oncology. In order to study further this phenomenon, we have developed an animal model of LC-induced granulocytosis. Four mongrel dogs weighing 23--28 kg were given commercially available LC in 300 mg capsules by mouth twice daily. Peripheral blood was examined twice weekly and marrow granylocyte-committed colony forming units (CFU-c) were studied weekly. Significant increments of peripheral granulocytes (P = 0.003) and of marrow CFU-c (P =0.001) were noted. An increased number of marrow CFU-c has not been previously reported but is consistent with other knowledge about this phenomenon. We conclude that this convenient animal model may be of value in further studies of LC-induced granulocytosis and its possible applications.


Acid phosphatase in normal human blood cells: characterization by isoelectric focusing in polyacrylamide gel. Using the high resolution technique of isoelectric focusing in polyacrylamide gel, isoenzymatic components could be discerned in acid phosphatase extracted from normal human lymphocytes, granulocytes, monocytes, and platelets. For each cell type, distinctive isoenzymatic patterns of acid phosphatase activity could be observed. These studies provide further evidence that various types of cellular differentiation are associated with differences in isoenzymatic components of acid phosphatase.