[Inhibiting effect of low density lipoproteins (LDL) on neutrophil function (author's transl)].

LDL preparations obtained from normal individuals of known genetic type have been added to neutrophils from healthy donors. After incubation at 37 degrees C for 30 min, and washing in Hank's solution, in order to evaluate metabolic and bactericidal activities neutrophils were assayed by the nitroblue tetrazolium (NBT) quantitative test, using latex particles, and by E. coli killing test. LDL in aliquots of 100-500 microgram/2 x 10(6) neutrophils, inhibited NBT reduction and reduced neutrophil killing of E. coli by more than 50%. No difference was observed in the degree of inhibition using LDL and neutrophils obtained from donors of either the same or of different Ag phenotype. These data suggest that the immunoregulatory activity of LDL affects lymphocyte as well as neutrophil function.


Na+/K+-dependent adenosinetriphosphatase in rabbit kidney. Separation by affinity chromatography.

We attempted separations by affinity chromatography of the Na+/K+-dependent adenosinetriphosphatase, using Sepharose 6B covalently coupled with ouabain, either on microsomal fractions or on preparations purified by discontinuous sucrose density gradient ultracentrifugation. The material, specifically eluted with ouabain, was tested to evaluate the recovery of ATPase activity and ouabain binding capacity. The results have shown the efficiency of this technique for (Na+/K+)-APTase isolation.


[Neurovegetative regulation of gastrin secretion].

The literature is reviewed in an account of the part played by the adrenergic and cholinergic systems in the regulation of gastrin secretion. As matters now stand, it can be said that, whereas the adrenergic system is capable of activating such secretion, the cholinergic system appears to be capable of modulating it by inhibition rather than stimulation. While it can be agreed that mediation of the adrenergic function occurs via the beta receptors, the possibility that the alpha receptors may also be responsible under certain conditions cannot be ruled out.


[In-vitro evaluation of morphine dependence. I. Response of the rat "fundus" to ACh].

The kinetic of cholinergic receptor systems was studied in an in vitro model for opiate tolerance and dependence. The fundus strip of rats chronically treated with morphine and sacrificed at different time intervals and in different conditions (with and without abstinence signs) was used. In evaluating the responses of fundus to ACh, the cholinergic receptor system showed the same response both in a condition of abstinence and in a condition of non-abstinence in vivo and in vitro. The response does vary however, according to the duration of the treatment: after an initial increase it comes back to its normal values, thus reproducing the tolerance pattern.


[Effect of chlorpropamide and of phenformin on isolated liver plasmatic membranes].

Previous observations from this laboratory suggest that chlorpropamide and phenformin, two hypoglycemic drugs belonging to sulfonylureas and biguanides respectively, act on liver plasma membranes altering the activity of enzymes bound to plasma membrane as (Na+-K+)-ATPase. This enzyme, an integral membrane protein which shows a cooperative behavior, has been used as an allosteric probe able to give information on plasma membrane. Our experiments show that (Na+-K+)-ATPase has a positive cooperative behaviour, as suggested from Hill coefficient n > l. The two hypoglycemic drugs decrease the Hill coefficient; in addition both chlorpropamide and phenformin inhibit (Na+-K+)-dependent but not Mg2+-dependent ATPase activity.


Relationship between thiamin phosphorylation and intestinal transport in vitro: effect of chloroethylthiamin.

The effect of Chloroethylthiamin (CET), a structural analog of thiamin, on thiamin pyrophosphokinase (TPKase) activity of the supernatant of rat isolated enterocytes was compared with that on thiamin intestinal transport by rat everted jejunal sacs. Thiamin- thiazol-2(14C) was used as a substrate both for TPKase activity and for thiamin serosal transport and uptake. CET strongly inhibited TPKase activity of isolated enterocytes: at molar concentrations 10 or 100 times higher than labeled thiamin, the inhibition was 57 and 100% respectively. The inhibition was of the competitive type, with a Ki = 15 microM. At a molar concentration 10 times higher than labeled thiamin, CET lowered the thiamin serosal transport by 60%, and the sac wall content of free and phosphorylated thiamin by 54 and 42% respectively. At the 1:10 thiamin: CET molar ratio, the extent of the reductions of TPKase activity (57%) and of phosphorylated thiamin content of intestinal sac walls (42%) were of the same order. This indicates a relationship between the two events. Moreover, since TPKase activity inhibition alone resulted in the lowering of labeled thiamin serosal transport, thiamin phosphorylation and transport are probably two strictly related processes.


Molecular mechanism of general anesthesia: II. Spin label studies on synaptic membranes.

In this communication we report the effects of general anesthetics on the mobility and order of spin labeled stearic acid derivatives in synaptic membranes and in bilayers formed from the lipids extracted therefrom. The anesthetics studied abolish the immobilization induced by synaptic membrane proteins on the membrane lipids : this effect, observed particularly in the bilayer core, is interpreted as a labilization of lipid-protein interactions induced by anesthetics.


Cerebellar malformations: some pathogenetic considerations.

1) Destructive processes are responsible for most cases of cerebellar microgyria of the trabecular pattern. Erosion and subsequent fusion of the folia produce the disorganized pattern in which the various cellular elements retain their noraml relationship and are capable of normal maturation. Intrauterine infection is responsible for most cases; the evidence is conclusive in some cases, presumptive in others. 2) Faulty genetic coding, as illustrated by the trisomies, may lead to formation of heterotopias. The primitive cells aggregating around the dentate nucleus should be interpreted as matrix cells and not as cells of the external granular layer. Cortical heterotopias with attempted internal organisation also occur; their origin is obscure. The unusual, possibly unique, transposition of the internal granular and Purkinje cell layers observed in one case may be ascribed to faulty formation of the Bergmann glia by analogy with the weaver mouse. 3) It is impossible at present to disentangle the role of genetic and environmental factors in the pathogenesis of the hysraphic malformations. It is possible, however, that defective fusion of the intraventricular cerebellar primordium plays a part in the development of the Dandy-Walker malformation, of midine cerebellar clefts in some cases of occipital encephalocele, and of extra-axial ependymal cysts of the posterior fossa.


Hypoglycaemia secondary to pancreatic islet cell adenoma.

The detailed case histories of 5 patients with hypoglycaemic episodes secondary to islet cell adenoma of the pancreas are presented. Clinical recognition of this syndrome remains the major problem but a full and detailed medical history is usually strongly suggestive of the correct diagnosis. The clinical diagnosis is confirmed by the repeated demonstrated that: 1) Symptomatic episodes produced by fasting are in fact due to hypoglycaemia (plasma glucose level less than 2.5 mmol/litre) 2) Such episodes are relieved by glucose administration 3) Concomitant hyperinsulinaemia is present (serum insulin greater than 8 micro-units/ml in the fasting state). Surgical resection of the adenoma produces a complete cure but the identification of the lesion at operation may be difficult and preoperative means of accurate localisation may be needed.


Adrenal reactivity to ACTH age changes in silver foxes inheriting different defensive behaviors.

We have studied the influence of selection for behavior in female silver foxes upon their adrenal reactivity to ACTH. It has been found that in the first year of life, in females from a population selected for domesticated behavior (tame), there is no differentiated adrenal response to different doses of ACTH. In wild females during the first year of life, varying strength of adrenal response to large and small doses of ACTH is noted. During the second year of life, the differential adrenal response to different doses of ACTH is improved in females of both behavioral groups; the reactivity of tame second-year females to small doses of corticotrophin surpasses that of wild second-year animals. It is concluded that during ontogeny in domesticated silver fox females there is a retardation in the establishment of an adequate response to ACTH by the adrenals, as compared to nondomesticated females.


Motor fibre refractory period and motor conduction velocity range.

The absolute and relative motor refractory periods and the range of motor conduction velocity were investigated in 25 normal subjects and in 25 patients suffering from peripheral neuropathy. The refractory periods were examined by a collision method, which eliminates the effect of the first of the paired stimuli, thus allowing the effect of the second stimulus to be clearly recorded. The range of conduction velocity was also examined by a collision method, and the conduction times were corrected by subtracting the absolute and relative refractory periods from the figures given by this method. The maximum motor conduction velocities obtained with this technique closely corresponded with those found using conventional methods. The results show that the refactory periods are prolonged and the conduction velocities are reduced in peripheral neuropathies, especially of segmental demyelination type, but the range of conduction velocity is not necessarily increased. The failure to suppress fully the effect of the proximal stimulus by a distal stimulus is a phenomenon found in peripheral neuropathies.


Effect of tetrahydrocannabinol and ethidium bromide on DNA metabolism and embryogenesis in Volvox.

The effects of delta 9-tetrahydrocannabinol (THC) and ethidium bromide (EB) on the developmental life cycle and DNA metabolism of Volvox carteri have been investigated. THC, previously shown to interfere specifically with cytoplasmic DNA (cDNA) in this organism, was used at different concentrations and at different times during the life cycle. The morphological consequences observed were found to be dependent on the nature and time of treatment. This study also indicates that ethidium bromide induces degradatin of cDNA similar to that mediated by THC. However, unlike THC, it also causes the cessation of nuclear DNA synthesis. The consequences of EB treatment on morphological development are different from those observed with THC. A correlatin of these observations with the biochemical results presented suggests possible models in which the amounts and proportions of nuclear and cytoplasmic DNA play a role in the regulation of embryogenesis in this organism.


[Allergy due to castor bean meal (author's transl)].

A comprehensive review of the literature on allergy due to castor bean is given as it occurs by inhalation of dust particles in the environment of milk working with castor bean. Furthermore it may occur by handling of castor bean and of its expressed residuals after oil production. A case report is presented about a respiratory allergy caused by castor bean meal containing fertilizer used in culturing grass plats. Symptomes of increased dyspnoe following repeated exposures and a strongly positive skin test to an extract of castor bean have been observed. There is strong evidence that the castor bean has a high allergenic potency, requiring attention in allergy tests.


The effects of levamisole on some functions of mouse macrophages after in vitro and in vivo administration.

In vitro incubation of peritoneal macrophages from normal, peptone-stimulated mice with levamisole (1-100 microM) for 1 and 22 h had no effect on either phagocytosis of particulate material (sheep erythrocytes, zymosan) or cellular levels and release of lysosomal enzymes (beta-D-glucuronidase, cathepsin D). By contrast, levamisole 1 and 5 mM dramatically increased enzyme release while inhibiting phagocytosis. In some experiments, however, these high concentrations of levamisole caused an elevated cell mortality. When incubation was extended to 72 h, a decrease of both phagocytosis and enzyme release was observed. The catabolism of endocytosed antigens (sheep erythrocytes, human gammaglobulin) was not at all or only slightly modified depending upon the antigen. The cellular level of cyclic AMP remained unchanged in all experiments. In vivo exposure of macrophages to levamisole (2.5 and 10 mg/kg/day i.p. for 3 days) produced a dose-dependent increase in processing of endocytosed antigens as shown by an enhanced transfer of initially endocytosed material to the macrophage plasma membrane. The other parameters were not modified. The immunogenicity of erythrocytes, when endocytosed by levamisole-treated macrophages and transferred into unsensitized recipients, was increased in some in vivo experiments.


Inhibition of human peptidyl dipeptidase (angiotensin I converting enzyme: kininase II) by human serum albumin and its fragments.

Purified peptidyl dipeptidase (angiotensin I converting enzyme or kininase II) from human lung or hog kidney is inhibited by commercially prepared plasma protein preparations, by human serum albumin and by the additive albumin stabilizer, acetyltryptophan. After the initial steps of purification, albumin was detected by immunodiffusion as a component in human lung peptidyl dipeptidase preparation. Fragment C of albumin (sequence 124-298) is a more potent inhibitor than the parent molecule (Ki = 1.7 X 10(-5)M). Reduction and carboxymethylation of five of the six S-S bridges in Fragment C yield the most potent noncompetitive inhibitor (Ki = 3 X 10(-6)M). Reduction of the sixth bridge raises the K1. This indicates that maintenance of the tertiary structure in Fragment C is of importance for the inhibition. Neither albumin nor Fragment C are substrates of the enzyme. Fragment C and its derivative also inhibit the inactivation of bradykinin by the purified human enzyme and by the peptidyl dipeptidase on the surface of intact cultured human endothelial cells.


Retroperitoneal lymph node dissection for Wilms' tumor.

Nephrectomy for Wilms' tumor was performed on 58 patients over a 20-yr period, and retroperitoneal lymphadenectomy was performed on 35 who presented with no demonstrable metastases. The survival rate was 100% for 19 clinical group I patients with negative nodes. Positive nodes in 9 instances led to 5 long-term survivors and significantly influenced staging as a guide for further therapy.


Nonvisualization of the intravenous pyelogram--a poor prognostic sign in Wilms' tumor?

In a series of 49 consecutive cases of Wilms' tumor from a single institution, there was a 20% incidence of nonvisualization on the initial intravenous pyelogram. Even though nonvisualization was secondary to either gross or microscopic invasion of the renal vein or renal pelvis, this did not forecast a grim progosis. Seven of the nine patients with nonvisualization are presently alive and tumor-free at least 5 yr after resection. The only two deaths in this particular group were unrelated to the Wilms' tumor.


Postradiation renovascular hypertension.

Radiation injury to arteries can represent a significant complication of therapeutic irradiation, even when the dosage used has not been excessive as judged by approved protocols. Children in whom therapeutic abdominal irradiation has been used should be monitored indefinitely for the development of hypertension. The presence of hypertension in such children with normal blood urea nitrogen (BUN) and creatinine, and without proteinuria, should prompt investigation for a renovascular lesion. Standard bypass procedures are usually effective, although the long-term success may be compromised by continuing changes in affected vessels.


[Eccrine poroma: its histogenesis and review of the literature].

It has been done an analysis of a case of eccrine porome, based on the morphology, considering the actual tecnical publications on the matter. The morphogenesis of the formation of this nevoid tumor is established by the continuity between the main body of the tumor and the underneath eccrine sweat glands. These glands present signs of hyperplasia, which in the highest levels present destructuration of the excretory duct and morphological modifications of the cells which are conditioned by the functional adaptation at different levels. In this way there is proliferation of structures, similar to the different parts of syringuium and acrosyringuium producing this hamartomatous neoformation.


Changes in psychological well-being during postmenopause as a result of estrogen therapy.

A study of the effects of a conjugated estrogen (Femipren) upon various climacteric symptoms, depression and anxiety scores and psycho-social factors in 120 women around the menopause, was carried out in a Menopause Clinic. In order to participate in this study, the women had to meet very strict criteria. A climacteric symptom list, the MMPI and a psycho-social questionnaire were used for assessment, and were employed before, during and at the end of 1 yr of hormone replacement therapy. Typical vasomotor and psychic symptoms changed significantly as did such factors as self-image, optimism, perception of the future and satisfaction with personal appearance, in those patients who received hormonal replacement therapy for 1 yr. As the psychological factors changed only in those patients who received long-term estrogen therapy, a tentative hypothesis of direct psychotropic effect of estrogen was formulated.


Adenosine triphosphatase of mycobacteria.

A Mg+2-(Ca+2)-activated adenosine triphosphatase activity has been demonstrated in saprophytic, human pathogenic and non-pathogenic, and atypical, species of mycobacteria. (Na+ + K+)-activated adenosine triphosphatase was absent in the species investigated. The effect of age of culture on enzyme activity was revealed in an increase up to mid logarithmic phase of growth, and a decline thereafter. Differences in oxygen tensin during growth did not alter enzyme activity. Isonicotinic acid hydrazide and streptomycin inhibited the enzyme activity.


Expression of the chromosomal mouse Beta maj-globin gene cloned in SV40.

The complete chromosomal mouse Beta maj-globin gene, including its intervening and flanking sequences, has been cloned in the monkey virus SV40. the mouse gene is transcribed, processed and translated in infected monkey kidney cells to yield mouse Beta maj-globin.


Identified neurones isolated from leech CNS make selective connections in culture.

Neurones cultured in vitro offer distinct advantages for studying how processes grow towards their targets and form synaptic connections. In contrast to the complex events occurring during the development of the nervous system, synapse formation in culture can be analysed in a few neurones at a time and under controlled conditions. We have now dissected out and cultured single identified neurones from the central nervous system (CNS) of the adult leech. Various types of sensory cells, motor cells, and interneurones can be identified in leech ganglia--each with a stereotyped set of properties, including: (1) the electrical characteristics of its membrane, (2) the arborisation of its branches and the morphology of its terminals and (3) the pattern of connections it makes with other identified neurones, skin or muscle. Thus, cultured cells can be compared in detail with their counterparts in situ. We have found that isolated cells survive for several weeks, maintain their membrane properties, sprout and form selective connections.


A physiological correlate of disuse-induced sprouting at the neuromuscular junction.

Recent investigations have established that many of the normal properties of muscle fibres are maintained, at least in part, by muscle activity. Thus, a fall in resting membrane potential, an increase in input resistance, and spread of acetylcholine receptors to extrajunctional sites can all be induced by abolishing muscle activity and prevented by direct stimulation of denervated muscle fibres. Muscle activity also exerts a trophic influence on the innervating motoneurones; furthermore it may be a factor in the regulation of sprouting. Brown and Ironton found fine, "ultra-terminal sprouts" emanating from the endplates of muscles rendered inactive by chronic conduction block of the muscle nerve. Pestronk and Drachman saw increased branching of the motor nerve terminal and a consequent increase in endplate size in similar conditions. If these sprouts at the endplates of inactive muscles were functional, one might expect more transmitter to be released in response to nerve stimulation. We report here that both quantum content and spontaneous miniature endplate potential (m.e.p.p) frequency are increased at the terminals of inactive (disused) muscles.


Epidermal growth factor receptors increase during the differentiation of embryonal carcinoma cells.

Mouse teratocarcinoma stem cells (embryonal carcinoma, or EC cells) bind very small amounts of mouse epidermal growth factor (EGF) and the latter hormone seems to have no stimulatory effect on the growth of two cloned lines of EC cells. However, when EC cells are induced to differentiate into large flat endodern-like cells (END cells), EGF receptors increase in number reaching a plateau in 6 to 8 days. At 8 to 10 days after induction, END cells multiply very slowly, but when EGF is added (3 x 10(-10) M) to the medium, cell division is stimulated and a further change in morphology occurs. This letter describes the binding characteristics and numbers of the EGF receptors on EC and END cells and shows that exogenous retinoic acid increases the numbers of EGF receptors on END cells. We were unable to find endogenous competing factors produced by EC cells. Such factors could account for the lack of detectable binding of EGF on these cells. As EC cells differentiate to END cells, so the ability of the cells to form tumours is reduced. Since this change is accompanied by an increase in the number of EGF receptors there may be a relationship between these two events.


Clindamycin and lincomycin alter miniature endplate current decay.

Antibiotic-induced muscle paralysis has frequently been found in both experimental animals and man with three distinct classes of antibiotic: (1) streptomycin and related aminoglycoside compounds, (2) polymyxins and (3) tetracyclines. Recently lincomycin and its chemical congener, clindamycin, have been reported to produce muscle paralysis which has different characteristics from those seen with other classes of antibiotic. Although closely related in chemical structure, lincomycin and clindamycin also seem to produce muscle paralysis by different mechanisms. Clindamycin is considered to exert a direct depressant action on muscle contractility whereas the action of lincomycin is considered to be primarily a depression of neuromuscular transmission. We report here that each of these antibiotics had a significant but different influence on endplate channel behaviour. Clindamycin increased the rate of miniature endplate current (m.e.p.c.) decay and reduced its voltage sensitivity without altering its exponential nature. Lincomycin split m.e.p.c. decay into an initial rapid phase followed by a prolonged phase.


Role of intracellular Ca2+ sequestration in beta-adrenergic relaxation of a smooth muscle.

Various mechanisms have been proposed for beta-adrenergically mediated relaxation of smooth muscle. All theories suggest the involvement of cyclic AMP as a second messenger: beta-agonists stimulate adenylate cyclase which converts ATP to cyclic AMP and protein kinase, activated by cyclic AMP, is then thought to catalyse a protein phosphorylation that leads to a reduction in free Ca2+, thus effecting relaxation. How this last step is accomplished is much debated, but the following possibilities are currently considered as the mechanisms responsible for cyclic AMP-induced reduction of cytoplasmic Ca2+: activation of a Ca2+-ATPase in the plasma and/or sarcoplasmic reticulum membranes which lowers cytoplasmic [Ca2+] in a direct manner or stimulation of (Na+-K+)ATPase in the cell membrane which may indirectly effect Ca2+ extrusion. Among the hypotheses suggested, those of Ca2+ sequestration by the sarcoplasmic reticulum and of Ca2+ extrusion across the cell membrane are consistent with each other if it is assumed that both processes are effected by a cyclic AMP-sensitive Ca2+-ATPase. However, quite a different mechanism is implied by involving the Na+-K+ pump and Na+-Ca2+ exchange carrier. In this report, we present evidence that suggests intracellular Ca2+ sequestration is the mechanism involved.


Extracellular but not intracellular application of peptide hormones activates pancreatic acinar cells.

Peptide hormones, like neurotransmitters, are traditionally thought to activate cells by interacting with receptor sites accessible only from the extracellular space. However, there is no available evidence that establishes whether intracellular injections of peptide secretagogues can or cannot initiate cell activation. In view of recent demonstration that peptide hormones can penetrate the intracellular space in some tissues and the reports that intracellular injections of the neurotransmitter, dopamine, into acinar cells of cockroach salivary gland cause stimulation it seems of fundamental importance to test directly whether introduction of peptide secretagogues inside acinar cells of mammalian exocrine tissue can induce cell activation without first interacting with the outer surface of the external cell membrane. The data presented here show that injections of the secretagogue peptides caerulein and bombesinnonapeptide (bombesin-NP) into pancreatic acinar cells fail to evoke the characteristic potential and conductance changes that are observed following extracellular applications of these peptides.


Correlation between benzodiazepine receptor occupation and anticonvulsant effects of diazepam.

The benzodiazepines are potent anticonvulsants for a wide variety of experimental and clinical seizure disorders. The demonstration of saturable, high-affinity and stereospecific binding sites for the benzodiazepines in the mammalian central nervous system suggests the presence of pharmacological receptors mediating the anticonvulsant properties of these compounds. The good correlation between the anticonvulsant potencies of a series of benzodiazepines and their ability to inhibit 3H-diazepam binding in vitro further supports this hypothesis, but evidence for a direct interaction between benzodiazepines and their receptors, and a subsequent inhibition of seizure activity (or elevation of seizure threshold) is lacking. Recent reports from our laboratory and others have demonstrated the feasibility of labelling benzodiazepine receptors in vivo following parental administration of tritiated benzodiazepine. This technique permits one to study the relationship between the anticonvulsant activity of the benzodiazepines in vivo and the number of 'drug-occupied' receptors in vitro. We now report that there is an excellent correlation between benzodiazepine receptor occupancy by diazepam and protection against pentylenetetrazol-induced seizures. Furthermore, these results demonstrate that only a small fraction of benzodiazepine receptors need be occupied to produce a complete anticonvulsant effect.


Glycyrrhizic acid inhibits virus growth and inactivates virus particles.

Screening investigations in antiviral action of plant extracts have revealed that a component of Glycyrrhiza glabra roots, found to be glycyrrhizie acid, is active against viruses. We report here that this drug inhibits growth and cytopathology of several unrelated DNA and RNA viruses, while not affecting cell activity and ability to replicate. In addition, glycyrrhizic acid inactivates herpes simplex virus particles irreversibly.


Effect of novobiocin on initiation of DNA replication in Bacillus subtilis.

The initiation of DNA replication of small replicons in vitro involves conformational changes in the whole DNA molecule or in the region near to the replication origin. One striking finding has been the role of DNA gyrase (that is, the necessity for supercoiled structure) in the initial stage of ColE1 replication in vitro. However, little is known about the effect of gyrase on the initiation of replication of bacterial chromosomes in vivo. We have constructed a map of cleavage sites of restriction enzymes at the region of the origin of replication of the Bacillus subtilis chromosome (accompanying paper). This has now enabled us to examine the effect of novobiocin, a selective inhibitor of DNA gyrase, on the replication of the specific chromosomal segments near the origin and to seek a possible role for the gyrase in the initiation of chromosomal replication. We have found that only a limited segment of the chromosome at the origin region was replicated in the presence of novobiocin. This effect allowed us to locate the site of the origin of replication to within a DNA fragment of molecular weight 3.4 x 10(6).


Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line.

A significant aspect of primary hepatic carcinoma in man is the high positive correlation of hepatocellular carcinoma with infection with hepatitis B virus (HBV)1. Analysis of the relationship between HBV infection and oncogenesis is difficult because natural infection with HBV is limited to man and experimental infection has been achieved only in chimpanzees and gibbons. Furthermore, because HBV has not been successfully propagated in cell culture, basic study of virus-cell interaction of the aetiological agent of one of the most widespread infections of man has been impossible. Recently, however, a cell line (PLC/PRF/5) derived from a human hepatoma biopsy was described which produces the HRV surface antigen (HBsAg) and so provides a tool for the experimental investigation of HBV in viro. We now report the derivation and characterisation of two additional cell lines primary liver carcinomas. In contrast to the PLC/PRF/5 cell line, these cell lines retain the capacity to synthesise many human plasma proteins, including both albumin and alpha-fetoprotein (AFP). One of these lines also produces BHsAg. We also present evidence that HBsAg synthesis and secretion in this cell line are correlated with the growth state of the culture. This finding is in contrast to the continuous HBsAg production found in the PLC/PRF/5 cell line.


The isolation and cultivation of calf rotavirus in the Republic of South Africa.

Calf rotavirus was cultivated and propagated in tissue culture from faeces of 3-week-old calves suffering from severe diarrhoea. Criteria for viral involvement were: production of cytopathic effect in primary foetal calf kidney cells, specific fluorescence, and identification of the agent by means of electron microscopy. In a limited serological survey the majority of the cows on an infested farm were found to possess neutralizing antibodies to the local rotavirus strain.


Genetic and physiological characterization of temperature-sensitive mutants of bluetongue virus.

Complementation studies were carried out, using temperature-sensitive (t-s) mutants of blue-tongue virus (BTV). The results proved to be inconclusive as only low indices of complementation were obtained. No discrepancy was found between the previous classification of these mutants in 6 recombination classes and the complementation data recored. In general, the t-s mutants require a latent growth period of 16-20 h at 28 degrees C and maximum titres can be demonstrated 40-48 h post-infection. One mutant, (F211), however, consistently had a growth lag phase of 32 h. Mutants of the 6 recombination groups were further classified into 2 groups by temperature-shift studies. One calss of mutants expressed their t-s lesion prior to 24 h and the other class only after 24 h post-infection. Mutant F73 was found to be defective in its ability to synthesize ssRNA at a late stage in the replication cycle at the non-permissive temperature.


The nucleic acid and proteins of epizootic haemorrhagic disease virus.

Purified epizootic haemorrhagic disease virus (EHDV) was shown to contain 10 double-stranded RNA segments and a double-layered protein capsid with 4 major and 4 minor polypeptides. The virus differed from bluetongue virus (BTV), the orbivirus prototype, in that EHDV had an additional minor polypeptide component. This component, together with the major polypeptides P2 and P5, formed the outer capsid layer of the virus. The extra polypeptide apparently stabilizes this layer since, unlike BTV, EHDV was quite stable on CsCl gradients at both pH 7,0 and 8,0. EHD virions were found to have a density of 1,36 g/microliter, while particles without the outer capsid layer were isolated and had a density of 1,40 g/microliter. Two non-capsid polypeptides, P5A and P6A, were identified in addition to the 8 capsid polypeptides. Polypeptide P5A was synthesized in excess of all the others. There was little homology between the nucleic acids of EHDV and BTV with only 5-10% cross-hybridization. No hybrid double-stranded RNA segments were identified. We found by cross-immune precipitation that the major core polypeptides of the 2 viruses (P7 and P3) have common antigenic determinants.


Isolation and characterization of antibodies to Clostridium perfringens epsilon toxin from hyperimmune horse serum.

Antibodies against epsilon toxin were isolated from hyperimmune horse serum by affinity chromatography. Purified epsilon prototoxin covalently bound to Affigel 202 was used as immunosorbent, and antibodies were eluted with 6.0 M guanidine chloride. In a single run 80 mg of antibody could be recovered from a 20 microliter column of immunosorbent. The antibody was shown to belong to the IgG(T) class of immunoglobulins.


Further characterization of the t-s mutant F207 of bluetongue virus.

Temperature-shift experiments verified that the t-s lesion of BTV mutant F207 is expressed late in the replication-cycle, that is, at a stage when all virus components have already been synthesized. All viral polypeptides were indeed found in the soluble but not in the particulate fraction of cytoplasmic extracts from infected cultures grown at the non-permissive temperature. This suggests that the t-s lesion could be a defect in one or both of the polypeptides P2 and P5, which are respectively reduced in amount and absent from the latter fraction. Alternatively, the lesion could be an inability of the core particle to bind these 2 outer capsid polypeptides.


Development and suppression of a population of late-adhering macrophages in mouse malaria.

Changes in phagocytic and adherent cell numbers were compared during the course of infections of mice with Plasmodium yoelii (Py) and P. berghei (Pb) and in vaccinated mice challenged with homologous parasites. Nucleated cells in the spleen increased in number in Py-infected mice and were maximal at the time of recovery. The number of phagocytic cells increased in parallel, as did the number of blood leucocytes. Rates of increase were accelerated in vaccinated mice. Changes in Pb-infected mice resembled controls and blood leucocytes showed no consistent increase. In infected mice, the number of spleen and bone marrow cells which adhered to plastic rose above normal. At some stages of infection, cells which did not adhere in 24 h did so in 72 h. Such late-adhering cells, which resembled macrophages in morphology, were most numerous at the time of recovery. They appeared to be derived from monocyte precursors which matured in culture. Sometimes cells adherent at 24 h suppressed the development of the late-adhering population. Silica invactivated these suppressive macrophages but did not affect the precursors which developed into late-adhering cells. It is concluded that malarial infection stimulates the production of precursors of the macrophage-monocyte series and that their development is regulated by the presence of mature macrophages.


Muscarinic supersensitivity: a possible model for the sleep disturbance of primary depression?

The sleep changes induced in normal volunteers following the administration of scopolamine on 3 consecutive mornings resemble many of the abnormalities observed in the sleep of patients with primary depression: increased sleep latency and reduced rapid eye movement (REM) latency, total sleep time, and sleep efficiency. Furthermore, in a multivariate discriminant analysis--previously shown to distinguish the sleep records of depresed patients from those of normal controls and insomniac patients--the records from baseline nights were selected as normal and those after scopolamine as predominately depressed. Those observations suggest to us that muscarinic supersensitivity in normals may function as a pharmacological model for the sleep disturbances of depression.


Scopolamine-induced muscarinic supersensitivity in normal man: changes in sleep.

Scopolamine (6 microgram/kg) was administered on 3 consecutive mornings to normal human subjects. Sleep recordings obtained at night (when the central anticholinergic effect of the morning scopolamine was no longer present) indicated a significant reduction in latency to REM-sleep onset on the nights following the second and third injections. This effect is opposite to the direct pharmacological action of nighttime administration of scopolamine (i.e., prolongation of REM latency). In addition, total sleep time and sleep efficiency were reduced, and sleep latency was increased. Furthermore, scopolamine pretreatment on 2 consecutive mornings also potentiated the REM-inducing effect of arecoline, a central muscarinic agonist. These data are consistent with the development of cholinergic supersensitivity following cholinergic blockade.


EEG sleep and affective psychoses: I. Schizoaffective disorders.

The sleep electroencephalogram (EEG) was studied in 41 depressed inpatients. EEG sleep records were compared for two diagnostic subgroups; patients with psychotic depression (n = 29) or with schizoaffective disorders (n = 12). As was true in the previous pilot study, no major EEG sleep variables distinguished the patients with psychotic depression from those with schizoaffective disorders. These data are consistent with the theory that all psychotic depressive states may have certain common psychobiologic features such as shortened rapid eye movement (REM) sleep latency.


Changes in lymphocyte beta-adrenergic receptors in depression and mania.

The beta-adrenergic receptors were studied in vitro in lymphocytes obtained from patients with major affective disorders and controls. Specific L-[3H]-dihydroalprenolol binding was decreased in both depressed and manic patients compared to controls and euthymic patients. Isoproterenol-stimulated, but not prostaglandin El-stimulated, cyclic adenosine-3',5'-monophosphate production was decreased in manic and depressed patients. These results suggest decreased lymphocyte beta-receptor functioning in depression and mania. This decrease may be an index of changes in brain beta-receptors in mania and depression, or may simply reflect homeostatic regulation of peripheral beta-receptors in response to stress-induced increases in circulating catecholamines.


MAO activity and EEG sleep in primary depression.

Platelet monoamine oxidase (MAO) activity and electroencephalographic (EEG) sleep measures were examined in 56 drug-free hospitalized patients with primary depression as defined by the Research Diagnostic Criteria. The group included 35 females and 21 males with a mean age of 42.6 +/- 1.4 years. Platelet MAO and EEG sleep data were compared for the group as a whole and separately for the unipolar, bipolar, male, and female subgroups. No significant relationships could be demonstrated for the entire group or for the unipolar, male, or female subgroups. However, an inverse relationship between MAO activity and REM sleep percent was noted in the bipolar subgroup (p < 0.02). While changes in REM sleep have been relatively firmly established in primary depression, the relationship of MAO to depression and to REM sleep remains unclear.


Dopamine-mediated behavior produced by the enkephalin analogue FK 33-824.

Intraperitoneal injection of FK 33-824 produced apomorphine-like stereotyped behavior in rats. Antagonism of this stereotypy by naloxone and neuroleptics suggests that FK 33-824 can activate opiate and dopamine receptors in the brain. Because increased dopaminergic neuronal activity is thought to be involved in schizophrenia and dopamine-mediated stereotypy has been used as an animal model for this illness, these results are consistent with an involvement of endogenous opiate-like peptides in schizophrenia. This involvement provides a possible mechanism for the reported improvement in schizophrenic psychosis produced by naloxone.


Low dose fluphenazine decanoate in maintenance treatment of schizophrenia.

To test the clinical efficacy of low dose fluphenazine decanoate (1.25 mg to 5.0 mg biweekly), we carried out two separate experiments: (1) an open trial in 57 schizophrenic outpatients, lasting 6 months; (2) a double-blind, placebo-controlled discontinuation study in a subgroup of patients who maintained good remission throughout the entire 6-month open trial. The results suggest that lower doses of fluphenazine decanoate than those usually used may be effective in preventing psychotic relapse while keeping total cumulative dosage to a minimum.


Pertussis toxin: the cause of the harmful effects and prolonged immunity of whooping cough. A hypothesis.

The nature of the pathogenesis and of the prolonged immunity of whooping cough has not been clearly defined. The literature of Bordetella pertussis indicated that only the antigen that induces histamine sensitization, lymphocytosis, and other biological reactions in mice is the cause of the harmful effects and prolonged immunity of whooping cough. This antigen has the general characteristics of bacterial protein exotoxins that cause the harmful effects of infectious diseases such as diphtheria and tetanus. It is proposed that this antigen, which is histamine-sensitizing, lymphocyte-leukocyte-promoting, and islets-activating (HSF-LPF-IAP), be designated pertussis toxin. Agglutinogen, hemagglutinin, and heat-labile (at 56 C) and heat-stable (at 100 C) toxins are no doubt interrelated with the immunologic and/or toxic reactions of whooping cough. It appears that the first defense against the disease is the antibody that prevents adhesion of the bacteria to the cilia of the respiratory epithelium and that the second defense is the antitoxin against pertussis toxin (HSF-LPF-IAP).


Differential diagnosis of infant botulism.

Clinical investigations of infants hospitalized with botulism demonstrate a remarkable uniformity of complaints and physical findings. Constipation precedes a course of progressive weakness and cranial nerve dysfunction. Examination reveals hypotonia, hyporeflexia, and a variable pattern of involvement of the motor cranial nerves. Initial laboratory investigations should include electrodiagnostic tests, because findings of an incremental response to rapid, repetitive nerve stimulation and of brief, small-amplitude motor units on electromyography are virtually pathognomonic of botulism in the infant. Differential diagnosis includes disorders that may produce generalized depression of the central nervous system, such as septicemia, meningitis, metabolic disturbances, and intoxications. Specific involvement of the neuromuscular system includes acute polyneuropathies, diseases of the anterior horn cell, congenital myopathies or muscular dystrophy, and neonatal myasthenia gravis. Recent studies have expanded the clinical spectrum of infant botulism to include some cases of sudden infant death syndrome and otherwise nonspecific constipation.


The action of botulinal toxin.

Two areas of research on botulinal toxin are reviewed: (1) isolation and characterization of the toxin molecule and (2) the mechanism by which the toxin acts to paralyze transmission by cholingerrgic nerves. The various molecules of botulinal toxin (types A, B, D, E and F) have molecular weights of approximately 150,000. The toxins are composed of two subunits with molecular weights of approximately 100,000 and approximately 50,000, respectively. The subunits are linked by one or more disulfide bonds. The large-molecular-weight substance (approximately 150,000) is fully neurotoxic; neither subunit possesses neurotoxicity. Toxin-induced paralysis of cholingergic nerves involves three steps: (1) an initial binding step that involves an external receptor; (2) a translocation step during which the toxin molecule, or some portion of it, moves through the nerve membrane; and (3) a paralytic step during which the release of acetylcholine is blocked.


Processes of transport and absorption in the developing infant intestine.

An understanding of changes in the processes of absorption, detoxification, and intoxication in the intestine of infants, especially during the perinatal period, may aid in reaching an understanding of why the syndrome of infant botulism is restricted to the first six months of life. Some of the important metabolic changes associated with both the perinatal and the postnatal period of development are emphasized in these studies. A more complete understanding of the processes by which substances are absorbed into the intestine and then reexposed via the enterohepatic circulation may provide clues about why infant botulism occurs.


The prevention of pig-bel in Papua New Guinea.

In many Highlands hospitals pig-bel is the commonest cause of death in children over 12 months of age. In a double blind controlled trial in Sina Sina we have shown that Clostridium welchii type C beta toxoid (beta toxoid) protects against pig-bel (p < 0.02). Of 2,538 children given beta toxoid, two developed pig-bel in the following two years. Seventeen of 2,532 control children vaccinated with tetanus toxoid developed pig-bel. We suggest that beta toxoid be given to Highlands children at two, four and six months of age. Boosters may be needed. Health services in the Highlands need to be improved so that this vaccine can be effectively delivered.


Effects of murine cytomegalovirus infection on the immune response to a tumor allograft.

The purpose of this study was to compare the effects of murine cytomegalovirus infection introduced after an EL4 ascites tumor allograft with cytomegalovirus infection accompanying or preceding the allograft. Parameters that were measured included documentation to the host's immune system. Depressed immune response of splenocytes from mice infected at any time before assay was documented by decreased responsiveness to phytohemagglutinin, to lipopolysaccharide, and to an alloantigen in mixed lymphocyte culture. In contrast, animals infected after grafting had enhanced lymphocyte-mediated cytolysis (LMC), enhanced serum-mediated cytolysis (SMC), and larger spleens than did animals that were only grafted and animals that were infected before grafting. Neither a depressive nor an enhancing effect of virus administered after grafting was reflected in vivo in reduced or increased graft clearance. Nonspecific effects of virus increased LMC and SMC in vitro, but the primary effect of viral infection after grafting is immune depression.


The possible role of Epstein-Barr virus in rheumatoid arthritis.

The sera of patients with rheumatoid arthritis form a precipitate with an antigen present in the sonicate of a human B (bone marrow-derived) lymphoid cell line (WiL2). The antibody is distinct from rheumatoid factor. The antigen can be demonstrated by an immunofluorescence technique to be distributed within the nucleus of the WiL2 cell and shows a discrete speckled pattern of nuclear staining. The nuclear antigen is not detectable in organ extracts from many animal species or in human T (thymus-derived) lymphoid cell lines. The current evidence suggests that the nuclear antigen is detectable only in human B lymphoid cells infected with Epstein-Barr virus. It is not clear at present whether the nuclear antigen is a viral protein or a cell protein induced by the Epstein-Barr virus as a result of polyclonal B cell activation.


Clostridium welchii type C antitoxin in the treatment of "pig bel" (enteritis necroticans): a controlled trial in Papua New Guinea.

Enteritis necroticans remains a serious problem in the Highlands of Papua New Guinea. Earlier claims that Clostridium Welchii Type C antitoxin is effective in lowering mortality rates were subjected to a prospective controlled trial in 1972-73. Use of the antitoxin was found to be of no benefit in treatment of this disease.


Non-linear dynamic phenomena in open reconstituted enzyme systems.

The dynamic pattern of an open reconstituted in-vitro enzyme system containing phosphofructokinase, pyruvate kinase and adenylate kinase has been investigated. The approach is experimentally based on a stirred flow-through reactor. Stationary concentrations of phosphofructokinase and pyruvate kinase are maintained by entrapment in polyacrylamide gel particles. The results fit to a theoretical model based on the initial kinetic responses of the enzymes involved. An S-shaped steady state input characteristic for fructose 6-phosphate has been experimentally obtained.


Proteins of bovine leudemia virus: p. 15 is the major phosphorprotein.

Sodium dodecylsulfate-polyacrylamide gel electrophoresis of bovine leukemia virus (BLV) grown in fetal lamb kidney cells in the presence of 32P-phosphoric acid showed that the slightly basic 15000 D protein (pp 15) is the major phosphorylated component of the virus. This result further distinguishes BLV from mammalian type C viruses which possess acidic major phosphorproteins.


[Determination and postnatal development of fructokinase activity in swine liver].

Starting from the spectrophotometric method, described in ADELMAN et al., optimal reaction conditions for the measurement of fructokinase (ketohexokinase) in pig liver were systematically studied. It was necessary to increase the concentration of the substrate and further to lower the concentration of ATP for an optimal Mg: ATP-radio of 2:1. Using the optimized method fructokinase activity was determined in pig liver in relation to age, beginning from the last days of pregnancy to puberty. In liver of fetuses and newborn piglets during the first two days of life no or only a minute activity of the fructokinase was recorded. Therefore, the high level of fructose in fetal blood results from the inability of the fetus to metabolize fructose synthezised in the placenta or the fetal organs. At the end of the first week of life the activity of fructokinase was 10 times, after the second week 15-20 times higher than at birth. This high level remains constant during the suckling period and after weaning. For this reason, piglets after the first week of life are able to metabolize fructose and after the third week to form glucose and to release it into circulation. In adult pigs the activity of fructokinase in the liver decreases slightly. It corresponds-as in rat and human-to the elimination rate of experimentally applied fructose from the circulation. Therefore, this enzyme even in pigs is of significant importance for the utilization of fructose.


[Dynamic behavior of the adaptive control system, hypothalamus--anterior pituitary--adrenal cortex. Animal experimental study of the adequate system dynamics of the direct and indirect parameters of the adaptive control system].

The dynamic response of the adaptive closed-loop control system hypothalamus-anterior pituitary-adrenal cortex after application of short-time disturb pulses were examined experimentally in the pig. Blood sampling was performed biologically non-reactive by chronically implanted vascular catheters in the upper vena cava. A circadian biphase periodical dynamics characterizes the distrubance response of the adaptive control system, respectively, its direct and indirect parameters. After distrubance variables induced initial excess the plasma corticosteriods undershoot the basic level over a long period. Conversely, the dynamics of the eosinophils is determined through an initial eosinopenic phase and a subsequent typical overshoot. Periodical dynamic is absent in the steady state. The controller of the adaptive system does not show any spontaneous activity in the steady state without stress. The absence of an endogenic periodicity of the dependent parameters likewise gives evidence of an active oscillator. On the other hand, the controlling unit of the adaptive system constitutes a control response of circadian periodicty after activation by disturbance variables in pulse shape. The high damping of the control system terminates the transient state within 24 hours, and the corticosteriods and the eosinophils reach the steady state level. This control quality of the adaptive control system gains a high importance in the origination of the spontaneous circadian periodicity by corticosteriods and eosinophils and for the biological compensation of exogenous distrubance variables.


Sleep, psychophysiological functioning and learning processes in schizophrenia.

Sleep research has underlined how sleep is a specific functional moment inserted in the more complex system of circadian psychobiological rhythms. Many experimental data have evidenced that in particular REM sleep is involved in information processing and cognitive processes. The authors examine correlations between daytime cognitive performances and sleep patterns in a group of 11 schizophrenic patients, and in a control group made of 11 non-schizophrenic (borderline, depressed, character disorders) patients. Results show that there exists a relationship between REM sleep and cognitive activities, beyond nosographic partitions. They on the other side suggest caution in using sleep as a means of discrimination between schizophrenic and non-schizophrenic patients.


Electron microscopic examination of skin and conjunctival biopsy specimens in neuronal storage diseases.

Skin and conjunctival biopsy specimens from fourteen patients with neuronal storage diseases were investigated using an electron microscope. The diseases were Tay-Sachs disease, ceroid-lipofuscinosis (Jansky-Bielschowsky type), Niemann-Pick disease (type B), highly suspected adrenoleukodystrophy, I-cell disease, mucolipidosis of the beta-galactosidase deficient type, Hurler disease, Hunter disease and Morquio disease. This examination provided valuable diagnostic information on some neuronal storage diseases but not on Morquio disease or highly suspected adrenoleukodystrophy. False negative results may sometimes occur using this examination method. However, this examination suggests the usefulness of skin and conjunctival biopsy specimens as a diagnostic tool in some neuronal storage diseases.


[Studies of the presence of enzymes in various tissues of swine. 4. Studies of the Ca-, Mg-, and Na-K-ATPase activities in the homogenate and cell fractions of brain of fetuses and piglets].

Studies were conducted into the activities of calcium, magnesium, and sodium-potassium ATPase as well as into crude protein levels in homogenate and cell fractions of brain of two foetuses, aged 111 days, and of two piglets each, aged two days or nine weeks. The average brain-borne crude protein percentages were 6.55 in the foetuses, 7.25 in the two-day piglets, and 9.60 in the nine-week piglets. Calcium, magnesium, and sodium-potassium ATPase activities, related to one gram of fresh brain mass, were found to go up along with growing age. Highest relative percentages of sodium-potassium ATPase were recordable from homogenate and from the mitochondrial and microsomal fractions of the foetuses. Certain quantitative differences were obtained from using calcium, magnesium, sodium, and potassium ions to stimulate ATPase in the fractions of cell nuclei, mitochondria, and microsomes. The optimum temperature for magnesium and sodium-potassium ATPase in homogenate and in the mitochondrial fraction of brain was 45 degrees C.


[Results of histochemical and biochemical studies of uterus of gilts during the estrous cycle].

Alkaline phosphatase activity as well as lipid, polysaccharide, glycogen, and acid mucopolysaccharide levels in the uteri of 79 gilts were histochemically tested during the first three sexual cycles. Intra-cyclic alkaline phosphatase activity exhibited clearly pronounced variations in the surface epithelium, with maximum values reached in metoestrus, and moderate variations in the endometrial stroma, with maximum values reached in dioestrus. Cycle-dependent variations were recordable also from the glycogen and lipid levels in the surface and glandular epithelia and from the acid mucopolysaccharide levels in the endometrial stroma. The activity of alkaline phosphatase as well as the levels of glycogen and acid mucopolysaccharides during the first cycle still were lower than those in the second and third. Biochemical examinations of the endometrium revealed significant cycle-dependent variations in the activities of alkaline phosphatase, inorganic pyrophosphatase, and succinate-dehydrogenase as well as in the concentrations of soluble protein and glycogen, with maximum values being recordable on the tenth day of cycle. No significant intracyclic variations were recordable from the activities of acid phosphatase as well as of aspartate alanine-amino transferase.


[Circadian rhythm of poly A polymerase in rat liver nuclei].

Isolated nuclei from the liver of rats maintained under daily scheduled conditions of light (12 hrs) and food (8 hrs) showed a peak of Mn2+-dependent poly (A) polymerase activity 6 hrs after the beginning of the feeding period, as shown by DNA dependent RNA polymerase B. In fasting animals the peak of poly (A) polymerase occurs at 18.00.


[Circadian rhythm of poly A polymerase in chromatin and nucleoplasmatic extract of rat liver].

Chromatin preparations from the liver of rats accustomed to eating during the first 8 hrs of a daily 12 hrs dark period exhibit the same fluctuations of poly (A) polymerase as whole nuclei (a peak at 15.00 in fed rats, a peak at 18.00 in fasting ones). Nucleoplasmic activity was detectable only on addition of exogenous primer.


[Treatment of swine cell line with antibiotics: effect on growth kinetics and susceptibility to foot-and-mouth disease virus and to mycoplasma-like organisms].

Cell cultures treated with tylosin tartrate and kanamycin sulphate antibiotics were studied in relation to the cell growth rate, the susceptibility to the foot-and-mouth disease virus and to the microorganism eradication. These treatments did not affect the cell growth rate and the cell behavior to the viral infection. On the other hand, the decontamination of the intracytoplasmatic formas of mycoplasma-like organism was not observed.


[Effect of various methods of preparation of chromatin on the distribution of Mn2+-dependent poly-A-polymerase activity in subnuclear fractions of rat liver].

The distribution of Mn2+-dependent poly(A) polymerase activity between chromatin and nucleoplasmic fractions in rat liver nuclei strongly depends on the method followed for the chromatin preparation. Currently used methods containing high salt cause a loss in bound activity not accompanied by a correspondent increase in the "free" one.


[Isolation of bovine cell lines and susceptibility to foot-and-mouth disease virus].

Morphology and chromosomes of cell sublines derived from two new bovine kidney lines are reported. Cell susceptibility to the foot-and-mouth disease virus is discussed. One of the sublines showed epithelial-like cells, while the remainder, elonged fibroblastic-like cells. Most of them had a diploid number of chromosomes. From these sublines only one was sensible to the foot-and-mouth disease virus.


[Effect of administration of alpha-amanitine on Mn2+ - dependent poly-A-polymerase in rat liver nuclei].

Mn2+-dependent poly(A) polymerase activity tested in isolated rat liver nuclei is unchanged both in the absence and presence of exogenous poly(A) 30 min. after administration of a dose of alpha-amanitin that inhibits DNA-dependent RNA polymerase B. Longer times of treatment cause poly(A) polymerase to drop to 50% in the absence of poly(A) and to increase almost twice in its presence.


Hydrocephalus-dementia-complex in Paget's disease.

The clinical triad of gait disturbances, memory impairment and urinary incontinence is associated with a communicating hydrocephalus in the 'normal pressure hydrocephalus'-syndrome. The authors present a case of H.D.C. (hydrocephalus-dementia-complex) in Paget's disease with an identical syndrome, but with obstructive hydrocephalus, causing a triventricular dilatation. Today, each case of mental deterioration in Paget's disease, should be immediately observed and neurosurgical intervention kept in view. An X-ray of the skull, a CT-scan of the brain and a cisternography are performed as routine procedure. If there are indications of involvement of the basis of the skull, hydrocephalus and/or disturbed pattern of the tracer-migration around the convexities, associated with a certain degree of dementia, impaired gait or urinary incontinence, a ventriculo-subcutaneous drain should be inserted. If clinical improvement follows, a ventriculatrial shunt is indicated. The post-operative clinical outcome seems to be dependent on some clinical and technical factors. The most eventful outcome is observed in cases where the clinial triad, described in patients with a classical 'normal pressure hydrocephalus'-syndrome, is associated with an obstructive hydrocephalus, due to a stenosis of the Sylvian aqueduct.


Bilirubin excess, erythrophages and siderophages in differentiation of blood in cerebrospinal fluid.

Non-artificial blood admixture in cerebrospinal fluid (CSF) can be proven by demonstrating siderophages and 'bilirubin excess'. Bilirubin excess exists when the bilirubin concentration measured in the CSF exceeds the expected CSF bilirubin concentration. The expected CSF bilirubin concentration is calculated by multiplying the serum bilirubin concentration by the albumin quotient. Given a certain method of determination, a difference between the above-mentioned concentrations of less than 0.15 mumol/l must not be regarded as excess. Bilirubin excess was exclusively found in CSF with non-artificial blood admixture. Cytological and spectrophotometric studies are sufficient in screening for non-artificial blood admixture. When there are indications of an increased bilirubin concentration, calculations should be made in order to establish whether bilirubin excess exists. Lumbar puncture within 48 hours of the suspected haemorrhage should be avoided if possible.


Radiotherapy in brain-stem gliomas in children.

Twenty-one children with brain-stem glioma are discussed. Sixteen received radiotherapy and five were too sick to be irradiated. The mean survival was 3.4 weeks for the non-irradiated group and 8.7 months for the treated group, two of the latter being still alive five and ten months after diagnosis. From the literature and our cases the effect of radiotherapy in brain-stem glioma in childhood is not clear. Differences in patient-selection criteria and the mode and doses of irradiation between the reported series may account for the divergent results, but in our cases radiotherapy did not prolong survival as significantly as compared with the reported series. Elevated intracranial pressure is not rare in patients with a brain-stem glioma, as may be concluded from the present series and from the literature.


Why should we study the infant 'near miss for Sudden Infant Death'?

The use of 'near miss for Sudden Infant Death syndrome' infants as a model toward understanding the Sudden Infant Death syndrome has been questioned. Although there are numerous problems in delineating this patient population and defining events occurring during sleep, continuous polygraphic monitoring demonstrates potentially life-threatening events.


Relationship between prostaglandins and cyclic-AMP in the process of implantation in rats.

The effect of alloxan, a known inhibitor of adenylate cyclase, on the levels of prostaglandins E and F was investigated in pregnant rats. Animals were treated intraluminally with alloxan in normal saline or alloxan plus cyclic-AMP in normal saline or normal saline alone on day 4 of pregnancy. The uteri were collected on days 5 and 6 of pregnancy and the prostaglandins E and F were extracted and measured by a radioimmunoassay technique. In the alloxan treated rats, a significant reduction in PGE and PGF levels was found on days 5 and 6 of pregnancy as compared to untreated control group, whereas in the alloxan plus cyclic-AMP treated group the levels of PGE and PGF were comparable to those of the untreated control group. These findings suggest a possible relationship between prostaglandins and cyclic-AMP in the process of implantation in rats.


Thromboxane B2: a cardiodepressant of isolated rat hearts and inhibitor of sarcolemma Na+ - K+ stimulated ATPase activity.

The actions of thromboxane B2 on various parameters of cardiac performance were studied using the isolated perfused rat heart model. In concentrations from 100 pg/ml to 1 microgram/ml TXB2 significantly reduced the total generated myocardial contractile force. These changes were usually associated with an increase in the coronary perfusion pressure indicating an elevated coronary vascular resitance. Significant coronary pressure alterations were seen with TXB2 concentrations between 1 ng/ml and 1 microgram/ml. No significant changes were seen in either the resting tension or heart rate after TXB2 administration. However TXB2 (10 pg/ml to 10 ng/ml,, significantly reduced the amplitude of the electrical activity as observed in R wave changes of the surface electrocardiogram recording. In another series of experiments the action of TXB2 on rat heart sarcolemmal ATPase activity was studied. TXB2 significantly reduced the activity of the MG++ dependent - Na+ - K+ stimulated ATPase (Na+ - K+ ATPase) in these membrane preparations in concentrations from 10 ng/ml to 1 microgram/ml. Kinetic studies demonstrated that TXB2 reduced Vmax and increased the concentration required of ATP, Na+ and K+ for half-maximal enzyme activity. TXB2 did not inhibit either Ca++ or ouabain-induced depression of Na+ - K+ ATPase activity. The activity of either Mg++ or Ca++ - stimulated ATPase was not affected by TXB2. These results suggest possible important actions of TXB2 on rat heart activity which may be related to Na+ - K+ ATPase inhibition.


Effect of prostaglandins on the increased corticosterone output induced by caffeine in the rat.

The effect of prostaglandins on the increase of corticosterone output induced by caffeine was investigated in rats. PGE2 and some synthetic prostaglandins (11-deoxy-derivatives), which stimulate steroid output in vitro, enhanced the effect of caffeine. Pretreatment with indomethacin inhibited the effect of caffeine.


Antibody dependent cell-mediated cytotoxicity to herpes simplex virus in man: the influence of drugs on polymorphonuclear leucocyte and mononuclear effector cells.

We have shown that the function of the effector cells in antibody mediated cell dependent cytotoxicity (ADCC) can be reduced by the use of a number of drugs and prostaglandins. Both polymorphonuclear leucocyte and mononuclear effector cells were studied. Aminophylline and isoproterenol markedly reduced this activity. All the prostaglandins tested--PGA1, A2, E1 and E2 had a similar but more profound effect. At a dose of 100 mg/ml hydrocortisone had relatively little influence and neither ASA nor indomethacin had any effect on the effector cells tested. It is suggested that the effect of PGE1 in stimulating a recrudescent herpes simplex infection may be by reducing the efficacy of ADCC in host defence.


Secretory effect of prostaglandins on the rabbit lacrimal gland in vivo.

Fluid secretion from the cannulated excretory duct of the rabbit lacrimal gland was collected in microcapillary pipette at 10 min intervals. Secretion rate and electrolyte concentrations (Na+, K+ and Cl-) in the lacrimal fluid samples were determined before and after injections of small doses of prostaglandins (PGE1, PGE2, PGF1 alpha, PGF2 alpha, 15 epi-PGF2 alpha, PGF2 beta, PGA1 & PGA2) into the external maxillary artery. Only PGE1 had a dose-dependent stimulatory effect on the lacrimal gland secretion. The maximum secretion rate which was induced by PGE1 (25 micrograms/kg) was 7.2 +/- 0.9 microliters/10 min. Na+ concentration in the fluid secretion stimulated by PGE1 was increased; however, K+ and Cl- concentrations were not changed. Pretreatment with atropine (50 micrograms/kg, I.A.) failed to prevent the stimulatory effect of low doses of PGE1. On the other hand, the secretory responses were abolished when propanolol (3 mg/kg, I.A.) was administered prior to PGE1. The results suggest that fluid secretion of the rabbit lacrimal gland in response to PGE1 may be caused by excitation of the sympathetic ganglia and/or by direct action on beta-receptors in the acinar cells.


[Diagnostic value of serum angiotensin converting enzyme activity in sarcoidosis (author's transl)].

Serum angiotensin converting enzyme activity was measured in 60 blood donors and 100 patients with sarcoidosis histologically confirmed using a spectrophotometric assay with hippurylhistidylleucine as substrate. Sex related differences in the enzyme activity could not be observed. ACE was found to be markedly raised above the normal range (the mean of the control subjects plus 2 standard deviations) in 53 (58.2%) out of 91 untreated patients. The distribution of the enzyme activities was significantly different as compared to the control group. Nine patients with sarcoidosis receiving steroids had a mean level within the normal range. ACE level in stage II group tended to be more elevated compared to stage I, but this difference was not significant. Extrapulmonary sarcoidosis (with and without pulmonary involvement) appeared to be associated with especially high ACE activities. The distribution of the enzyme activities within the groups of untreated patients failed to reveal statistically significant differences. Therefore it was not possible to determine the clinical state of the disease and to discriminate between the stages (I and II) of sarcoidosis by the measurement of serum ACE level. However as the results suggest serum ACE assays can be a useful aid in diagnosis of sarcoidosis. Further investigations may demonstrate the value of this in vitro method for the clinical management of the disease including prognosis and therapeutic effects.


Self-mutilation and sleep stage in the Lesch-Nyhan syndrome.

Correlation of self-mutilation and sleep stage in the Lesch-Nyhan syndrome was studied by a polygraphic method. Five patients and three control boys were monitored with EEG, EOG, EMG, ECG and respiration throughout the night. The results were as follows: 1. The sleep-time of the patients was much disturbed during the night. 2. Decreased REM density was noticed with low DQ of the patients. 3. Self-mutilation during sleep-time was observed a lot in stages 1, 2, and REM in two cases. 4. No correlation was observed between body movement and self-mutilation in the Lesch-Nyhan syndrome. These data suggested functional disorders of the frontal lobe in the patients.


Pilot sero-epidemiological survey on Japanese encephalitis in north-western Burma.

A pilot sero-epidemiological survey of Japanese encephalitis was carried out in three areas of north-western Burma applying the hemagglutination-inhibition test. The existence of Japanese encephalitis infection in these areas was confirmed and its incidences in these areas were estimated and compared graphically.


Demonstration of cells with Marek's disease tumor-associated surface antigen in chicks infected with herpesvirus of turkey, O1 strain.

The presence of Marek's disease tumor-associated surface antigen (MATSA) was demonstrated by the direct and indirect membrane immunofluorescent tests, in chicks inoculated 7-10 days earlier with herpesvirus of turkeys (HVT), O1 strain. In in vitro cultures of spleen lymphocytes and ovaries obtained from these chicks, MATSA-positive cells were also detected after 1-7 days cultivation. A possible mechanism of protection by HVT vaccine against Marek's disease is proposed.


Photokinetic microassay of adenylate kinase using the firefly luciferase reaction.

A new rapid photokinetic method is described for determining the activity of adenylate kinase (ATP:AMP phosphotranspherase, EC 2.7.4.3) in 0.1--5.0 micrograms of freeze-dried tissue. This represents a weight range far below that obtainable by fine-needle biopsy. The reaction 2 ADP in equilibrium with AMP + ATP was employed and the ATP formed assayed with firefly luciferase as light yielder. The light emission was recorded on a multi-channel scaler. The adenylate kinase activities found in tissues of mice were in the same range as previously described in a study using fluorometric microassay.


The measurement of subnanosecond fluorescence decay of flavins using time-correlated photon counting and a mode-locked Ar ion laser.

A system is described consisting of a mode-locked Ar ion laser and time-resolved photon-counting electronics. The system is capable of measuring fluorescence lifetimes in the subnanosecond time domain. The Ar ion laser is suitable for the excitation of flavins, since the available laser wavelengths encompass the first absorption band of the yellow chromophore. Due to the high radiation density and the short pulse, both the time and wavelength resolution of the fluorescence of very weakly emitting compounds can be measured. Experiments have been described for flavin models exhibiting single and multiple modes of decay. In these examples lifetimes were determined both from deconvolved decay curves and from direct analysis of the tail of the curve, where no interference of the exciting pulse is encountered. Both determinations showed very good agreement. Due to the highly polarized laser light the decay of the emission anisotropy could be measured directly after the exciting pulse. In principle, fast rotational motions might be detected. An anisotropy measurement conducted with a flavoprotein with a noncovalently attached FAD is presented.


Small-scale techniques for the analysis of recombinant plasmids.

Using the cloning of part of the T-DNA of pTiC58 from Agrobacterium tumefaciens as an example, techniques are described which enable recombinant plasmids to be mapped and used as hybridization probes. In all cases the starting material is a colony of cells grown on an agar plate which is then subjected to lysis by lysozyme and Triton X-100 in volumes of the order of 300 microliters thus eliminating the need for handling and centrifuging liquid cultures under restrictive containment conditions.


Iatrogenic nondiabetic hyperosmolar states.

Four cases of the iatrogenic nondiabetic hyperosmolar state are presented. The clinical presentation, biochemical findings and management are discussed. No hypertonic solution should be infused at a rate above the level of patient tolerance; irrigation of a hollow viscus with a hypertonic solution should be avoided, and salt should not be used as an emetic. Patients under stress are particularly prone to this condition, largely because of the high circulating cortisol levels. The use of corticosteroids, salt-containing solutions in excess of patient requirements, water depletion and intravenous nutrition in the absence of careful biochemical monitoring, are all factors which may precipitate the hyperosmolar state in the critically ill.


Mechanism of beta-adrenergic relaxation of smooth muscle.

The mechanism of beta-adrenergic relaxation was investigated in isolated smooth muscle cells. Beta-adrenergic agents stimulate cyclic AMP-dependent phosphorylation, enhance Na+/K+ transport and induce relaxation. The stimulation of Na+/K+ transport is obligatory for relaxation, and we suggest that this stimulation induces relaxation through enhanced Na+/Ca2+ exchange.


Hormonal regulation of peptide receptors and target cell responses.

Regulation of plasma membrane receptors for peptide hormones by the prevailing ligand concentration often causes altered target cell function. Receptor number is determined by hormone-induced changes in membrane conformation, irreversible ligand binding, and processing of ligand-receptor complexes during hormone action.


Preoperative cell-mediated immune status of patients with malignant glial tumors.

The leukocyte adherence inhibition assay (LAI) was used to measure cell-mediated immunity in 26 patients with malignant glial neoplasms and in 41 control subjects. A significant inhibition of leukocyte adherence was observed in 21 out of 26 (80%) patients with malignant astrocytic gliomas in the presence of a 3M KC1 extract of glioma tissue compared to that of normal brain extract. Among the control group, no significant difference in the percentage of nonadherent leukocytes (NAL) was noted in the presence of either antigen. To study the specificity of the reaction, 3M KC1 extracts of meningioma, pituitary tumor, carcinoma of breast, carcinomas of lung, melanoma, brain, and heart tissues were employed as non-specific antigens. Such studies revealed significantly lower values of NAL. These data indicate that patients with malignant glial neoplasms manifest a cellularimmune response to glioma-associated antigens that can be measured by the tube LAI assay and that LAI assay may render additional useful information in the diagnostic and prognostic evaluation of malignant glial neoplasms.


BCNU and steroids in the viral-induced dog brain tumor.

Schmidt-Ruppin strain of Rous Sarcoma was inoculated intracerebrally into 27 newborn beagle dogs. Fourteen days after viral inoculation, 13 of the dogs were given intravenous BCNU (1 mg/kg). The other 14 were given the same volume of intravenous saline in a randomized, double-blinded fashion. Ninety percent of all dogs developed intracranial tumors. Radionuclide (mercury 197) brain scans were done on each dog at 2-week intervals. Median survival was 113 days in the BCNU group and 115 days in the placebo group (P > .99). Unequivocally positive radionuclide brain scans were detected in 5 dogs treated with BCNU and in 2 of the controls. There were no gross or microscopic differences at autopsy between treated and nontreated animals. BCNU, as given in this animal brain tumor model, did not demonstrate any oncolytic effect. An improvement in sequential brain scans was detected in 2 other dogs in response to Dexamethasone, which was given in a double-blinded, cross-over controlled fashion. Computerized tomography clearly demonstrated the tumor in two cases.


Isolation and identification of the Sersenk strain of goat pox virus in Iraq.

Goat pox virus was isolated during an outbreak of pox infection among goats in the Sersenk district, Iraq. The isolated virus grew on the chorioallantoic membranes of developing chick embryos and in primary lamb testis cell cultures. It was identified morphologically as a pox virus and serologically as a member of the Capripoxvirus group in the family Poxviridae. The isolated virus was designated the "Sersenk" strain.


Skin ulceration associated with herpesvirus infection in cats.

Three cases of skin ulceration in cats are described. Herpesvirus was isolated from the ulcers. Treatment of one case with antibiotics and an antiviral agent was successful. The significance of the viral isolate is discussed.


[A case of "sensitive deficiency of polyneuropathy"].

One case of carential sensitive neuropathy in a patient 47 years old with poor and imbalanced nutrition is reported. The clinical picture is characterized by mucocutaneous lesions, paresthesias like "burning feet", serious ataxia. All these symptoms remitted after one year of appropriate therapy. The carential vitaminic aethiology and the nosographic position of the disease are discussed on the basis of the current literature.


Experimental transmission of bovine papilloma virus (BPV) extracted from morphologically distinct teat and cutaneous lesions and the effects of inoculation of BPV transformed fetal bovine cells.

Bovine papilloma virus (BPV) was extracted from five cattle each affected with only one of five morphologically distinct lesion types. When inoculated into experimental calves either by scarification or intradermal injection, the BPV extracts produced lesions macroscopically and microscopically similar to those from which individual extracts were made. Fetal bovine cells, transformed in vitro with BPV, failed to produce fibromas, fibropapillomas or papillomas when inoculated into experimental calves. When calves inoculated with virus or BPV transformed cells were challenged with the five original BPV extracts, a differential immunity was demonstrated, while control calves were susceptible to all extracts. Post mortem examination revealed the presence of upper alimentary tract papillomas in three of eight calves forming one group. These results suggest that different strains of BVP, causing morphologically separable lesion types, exist. There may be additional BPV variants causing fibropapillomas of the teat and anogenital regions of cattle. The inoculation of BPV transformed fetal bovine cells conferred a relative immunity to later challenge with some but not all BPV extracts.


Human adenovirus antibody in sera of normal and tumour-bearing dogs.

Serum specimens from 42 normal dogs and 42 with untreated malignant tumours were assayed for the presence of antibodies to human adenovirus types 5, 21 and 31 and to infectious canine hepatitis (ICH) virus. Radioimmunoassays using human adenovirus antigens showed that 71 per cent (30/42) of all dogs with tumours, but only 19 per cent (8/42) of all normal dogs, were positive for human adenovirus antibody. Canine sera reactive with antigens of one human adenovirus type in radioimmunoassays were also reactive with antigens of the other two types. Dogs bearing malignant lymphoma or squamous cell carcinoma tumours had higher levels of antibody against adenovirus type 5 antigens. Human adenovirus type 5 was neutralised by sera from four tumour-bearing and two normal dogs, while sera from two normal and five tumour-bearing dogs were positive in immunodiffusion tests with human adenovirus antigens. Levels of ICH antibody in sera of normal adult dogs and adult dogs with tumours were not markedly different when measured by radioimmunoassays. Likewise, sera from these two groups of dogs had similar ranges of ICH neutralising antibody titres. In contrast, levels of ICH antibody detected by the serological assays in sera from non-pet, non-vaccinated pups were either markedly low or absent. Possible explanations for the observed increased levels of human adenovirus antibody in sera of tumour-bearing canine pets are discussed.


The demonstration of maedi/visna virus in sheep in Great Britain.

A virus indistinguishable from that causing maedi/visna has been isolated from a sheep flock in Great Britain. The virus was identified in cell cultures using immunofluorescent and electron microscopical techniques. The complement fixation test and the agar gel immunodiffusion test were used to assess the proportion of serological reactors within the flock. There has been no evidence of clinical disease in the flock into which foreign imports were introduced in the years prior to 1976.


Histological and histoenzymological studies of medulla oblongata of Taphozous melanopogon Temminck (a Microchiroptera).

The paper deals with the histological and histoenzymological studies of the medulla oblongata of Taphozous melanopogon Temminck (a microchiropteran bat). Three phosphatases, namely acid phosphatase (ACP), thiamine pyrophosphatase (TTP) and glucose-6-phosphatase (G-6-P) were studied. All the enzymes were seen in all the neurons. The large neurons of the different nuclei are more strongly positive for ACP than the small neurons. Further, the areas which contain dense populations of neurons are more strongly stained than the area containing scattered neurons. TPP and G-6-P distributions are almost parallel to that of ACP. The functions of these enzymes in synthesis and secretory processes were discussed.


Gastric carcinoma in dogs produced by the combined use of N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) and gastrin--with special reference to development of scirrhous carcinoma.

Eight beagle dogs of four-month-old received both oral administration of N-Ethyl-N'-Nitro-N-Nitrosoguanidine (ENNG) and subcutaneous injection of gastrin, and one of them was found to have an annularly infiltrating advanced carcinoma with marked fibrous thickening of the antral wall resulting in stenosis of the antrum (carcinoma scirrhosum) which resembled "Linitis plastica" (Borrmann's type IV carcinoma) in human stomach.


Adult T-cell leukemia with generalized cytomegalic inclusion disease and pneumocystis carinii pneumonia.

An autopsy case of adult T-cell leukemia with generalized cytomegalic inclusion disease and pneumocystis carinii pneumonia is reported. Tumor cells had T-cell characteristics (E-rosette) and cerebriform nucleus similar to Sèzary cells. Generalized lymphadenopathy, hepatosplenomegaly and an ectopic pancreas in the ileum were found at the time of autopsy. Histologically, leukemic infiltration was observed in almost every organ, and perivascular infiltration, vascular invasion were conspicuous findings. Cytomegalic inclusion bodies were observed in most organs (lungs, salivary glands, pancreas, liver, ectopic pancreas, sweat gland, stomach, thyroid gland, pituitary body, etc.). An acute hepatitis, probably caused by cytomegalovirus, was also noted. Presumed correlation of adult T-cell leukemia, cutaneous T-cell lymphoma and T-cell lymphoma was discussed.


Vascular leiomyoblastoma of the stomach -- Observations supporting its origin from Zimmermann's pericyte.

Gastric leiomyoblastoma with small mesenteric metastasis in a 44-year-old man has been studied by light and electron microscopy. Histologically, the tumor, having "epithelioid" tumor cells mixed with spindle leiomyomatous cells, was rich in blood vessels and focally simulated hemangiopericytoma. Electron microscopy revealed the tumor cells in variable stages of differentiation from poorly differentiated polygonal cells to smooth muscular cells. The intermediate cells had numerous cytoplasmic processes which interlocked each other. In addition, an intimate association was noted between the tumor cell processes and the small vessels including capillaries. With the recent characterization of the pericyte as a pluripotent mesenchymal cell that may serve as a precursor to the vascular smooth muscle cell, the present observations support the view that some leiomyoblastomas originate from the pericyte of Zimmermann. Leiomyoblastoma may be placed between hemangiopericytoma and glomus tumor in the spectrum of pericytic tumors.


Primitive neuroectodermal tumor (neuroepithelioma) of spinal nerve root -- Report of an adult case and establishment of a cell line.

A case of primitive neuroectodermal tumor arising in the cervical nerve root of a 28-year-old man is presented. Histologically, the tumor was characterised by proliferation of primitive neuroectodermal cells and formation of numerous Homer-Wright type rosettes. A cell line (Nagai line) was established from the tumor. Electron microscopic examination of Nagai cells revealed numerous microrosette formation with microvilli-like cytoplasmic processes projecting into the central lumina. Neurosecretory granules appeared in the cytoplasmic processes when Nagai cells were treated with dibutyryl cyclic AMP. Primitive satellite cells which completely surrounded other tumor cells with their tongue-like slender cytoplasmic processes were also found. Histogenesis of this unique tumor was discussed comparing with the neuroblastoma of sympathetic nervous system, medulloblastoma of the central nervous system, and with the tumors induced by Adenovirus type 12 in animals. It was concluded that the tumor was neuroepithelioma derived from a primitive stem cell of neural crest origin which possesses the bipotency to differentiate toward either neuroblastic or neurilemmal line.


Gaucher-like cells in juvenile GM1-gangliosidosis and in beta-thalassemia -- A histochemical and ultrastructural observation.

Peculiar storage cells appearing in bone marrow aspirates from a patient with juvenile GM1-gangliosidosis and from one with beta-thalassemia were examined light microscopically, histochemically and electron microscopically. Light microscopically, most of the storage cells closely resembled Gaucher cells pathognomonic for Gaucher's disease. The cytoplasm of the Gaucher-like cells contained numerous variable-shaped membrane-bound inclusions mostly arranged in a mosaic pattern and filled with fibrillar materials. Intermingled tubular structures were usually narrow as compared to those of the Gaucher cells. These ultrastructural differences of the stored materials between the Gaucher-like cells and Gaucher cells were more clearly substantiated by the high resolution electron microscopy with negative staining technique. Enzyme cytochemically, acid phosphatase activity was proved in or around the storage inclusions, suggesting their lysosomal origin. Histochemically, it might be suggested that the stored materials of the Gaucher-like cells in juvenile GMI-gangliosidosis were non-sulfated acid mucopolysaccharides and glycopeptides, whereas glycoproteins were the major component of the storage cells in beta-thalassemia. Possible mechanisms of storage in the Gaucher-like cells were discussed in both disorders.


The sensitivity of phage DNA and plasmid DNA for a restriction enzyme for Staphylococcus aureus.

In Staphylococcus aureus transduction of different tetracycline and chloramphenicol plasmids with a group I/III modification was possible to group I and III strains. Group II strains, containing a restriction endonuclease, had a restriction both for the phage and the plasmids: two restriction-deficient group II strains were good acceptors for these plasmids.


Chemical and electron microscopic studies of factors associated with the release of penicillinase from Staphylococcus aureus.

The effect of various factors such as sodium chloride, sodium citrate, pH, linase by Staphylococcus aureus ATCC 14458 was investigated. Penicillinase was measured at selected time intervals from supernates of cultures grown in Antibiotic Medium 3 broth containing various concentrations of salts or buffers or from supernates of cultures treated with lysostaphin and subsequently disrupted by French press treatment. Incubation of cells with media containing either sodium chloride (5, 10, and 15%), sodium citrate (5 and 10%), or organic buffers (Tris-HCl, 2.5, 5.0, and 7.5%; BES, 10 and 20%) resulted in a significant stimulation of the release of penicillinase when compared to control cells. It was also observed that pH 7.0-7.5 was optimal for penicillinase activity and release. From studies of enzymatic and mechanical disruption of cells, it was observed that an increase in ionic strength of the suspending medium to certain optimal levels appeared to stimulate the conversion of penicillinase to an extracellular form. Electron microscopic studies revealed that a large number of mesosomal vesicles seemed to be present in cells incubated for 4 hours in media containing various concentrations of sodium chloride. It is proposed that either appearance of vesicles or convolution of cell membrane, which may be caused by further synthesis of new membrane, is involved in stimulation of the synthesis and release of membrane-bound penicillinase.


Regulation of collagen synthesis and maturation by 3,4-dehydroproline.

Proline analogs are readily incorporated into collagen and noncollagen proteins. Since the imino acid content of collagen is greater than other proteins, it is suggested that the incorporation of a proline analog into cellular protein would have a maximal effect on collagen metabolism. Using a partially purified amino acyl tRNA synthetase preparation, various proline analogs were tested for their ability to inhibit Pro-tRNA synthesis. Amongst those tested, dehydroproline was the preferred inhibitor. Dehydroproline was also a substrate for amino acyl tRNA synthetase. When dehydroproline was added in vitro to membrane bound polysomes, the synthesis of collagenous proteins was preferentially inhibited. The addition of dehydroproline to mammalian cell cultures caused a marked reduction in prolyl hydroxylase activity. Under these conditions growth of cells, activities of lysyl; hydroxylase or lactic dehydrogenase were not affected. Reduction of prolyl hydroxylase activity by dehydroproline required protein synthesis. Removal of dehydroproline from the growth medium resulted in an increase in prolylhydroxylase activity. Hepatic fibrosis can be induced in rats by chronic administration of carbon tetrachloride. Under these conditions, the collagen content and prolyl hydroxylase activity of the liver is enhanced. Treatment of these fibrotic animals with dehydroproline results in a reduction of prolyl hydroxylase activity of the liver. A mechanism by which dehydroproline reduces prolyl hydroxylase activity will be discussed. Since prolyl hydroxylase plays a key role in the maturation and deposition of collagen, specific inhibitors of this enzyme are potentially useful in controlling collagen deposition in various pathological conditions.


Induction of germination in Blastocladiella emersonii by cyclic AMP and inhibitors of cyclic AMP phosphodiesterase.

Since K+-induced germination of Blastocladiella emersonii is accompanied by a rapid decrease of a specific cyclic AMP phosphodiesterase (cAMP PDE) activity and a transient cyclic AMP accumulation, the effects of this compound as well as of some inhibitors of cAMP PDE on the induction of germination were tested. Adenine and caffeine, competitive inhibitors of zoospore cAMP PDE, were able to elicit germination in substitution for K+. Cyclic AMP is a poor inducer, but a synergistic effect was evident when non-effective concentrations of K+ and cyclic AMP were added together to the medium. At the same concentration, cyclic GMP had no effect as compared with cyclic AMP. Lanthanum, a specific antagonist of calcium in several biological systems, completely blocked the germination induced by potassium.


Role of hemolytic factors of Sendai virus on human erythrocyte membrane fluidity.

In order to establish whether the hemolytic activity of Sendai virus might be related to changes of the membrane fluidity of sensitive cells, membrane fluidity variations have been recorded in human erythrocytes ghosts in the presence of Sendai and PR 8 viral particles. A decrease of membrane fluidity is elicited by Sendai and Pr 8 virions and by non hemolytic Sendai virus after trypsinization; the effect on membrane fluidity of sendai virus is strongly reduced after pyridine treatment which abolishes hemolytic action.


Identification of a glycoprotein inhibitor of human serum, active towards the hemagglutinating activity of Sendai virus.

Non-antibody hemagglutination-inhibiting activity of human serum is due, at least partially, to a glycoprotein inhibitor. This inhibitor was obtained from the whole serum or the fraction VI (Cohn) by means of methods of purification generally used for the preparation of the orosomucoid and was identified more precisely with an acid alpha 1-glycoprotein.


Decline in incidence of hepatitis A infection in Milan. A serologic study.

The availability of serum samples collected in 1958 and in 1977 from healthy individuals of Milan, has provided an opportunity to compare the prevalence of anti-HAV and anti-HBs in the same area at a distance of about 20 years. The results obtained show a general drop of anti-HAV rate from 1958 to 1977. Difference in antibody prevalence was found to be significantly remarkable (35.7% versus 3.6%, P < 0.05 and 78.6% versus 26.9%, P < 0.01) in the youngest age groups examined (1-5 and 6-15 years old). Moreover the plateau of antibody acquisition was reached over 30 years of age in people selected in 1958 and over 50 years in 1977. No substantial difference was found in anti-HBs prevalence in sera collected in 1958 and in 1977 (29.2% versus 29.3%). The finding of a decline in hepatitis A infection may reflect the improvements in public health and hygienic conditions which occurred in our city in the last decades, whereas with regard to hepatitis B, the maintenance of the same incidence of infection may be due to different contributory factors.


[Observations on the diffusion of hepatitis A and B virus during an epidemic in a Sicilian town with a high rate of infection].

Epidemiological investigations were carried out during a viral hepatitis outbreak occurring in a Sicilian town of 30,000 inhabitants with poor sanitary standards, with the aim to study the mode of spread of HAV and HBV in conditions of high incidence of infections. HBsAg, anti HBs, anti HAV (RIA), HBeAg, anti HBe and anti HBc were investigated in serum samples from patients, their family contacts and from healthy individuals of different age groups. Morbidity was inferred from case notifications; search of unreported cases among school children, through the study of absenteeism, did not reveal further cases. In all 148 cases, occurred from August 1976 through July 1977 with a peak in January, 44% were under 5 and 93% under 10 years of age. All but 8 of 59 cases in which laboratory data were available were due to HAV. Anti HAV antibodies were highly prevalent in serum samples obtained in February through April 1977: 62% were positive in the 1 to 3 years age group, and more than 90% among school children. Prevalence of HBsAg was age and sex dependent, ranging from 4% to 15%; anti HBs was present in 8% of the children 1-10 years and in 30% or more in age groups 30-41 and over. It is suggested that direct contact between very young children was the main mode of spread of HAV, and inapparent cases the main source of infection, although ambient diffusion through water contamination could not ruled out. HBV was probably propagated mostly by intrafamilial spread with little overt pathology.