Thyroxine-induced cardiomegaly: assessment of nucleic acid, protein content and myosin ATPase of rat heart.

1 mg/kg L-thyroxine was administered to rats for 14 days to evaluate the potential of the hyperthyroid state to induce heart hypertrophy and its effect on myosin adenosine-triphosphatase (ATPase) activity. Evidence of hyperthyroidism such as weight loss, elevation of rectal temperature, increased heart rate and oxygen consumption, was observed in all treated rats. Cardiac enlargement was determined by comparison of wet and dry ventricle weights, myocardial RNA, DNA and protein content. Wet and dry ventricle weights and the level of cardiac RNA and protein were augmented by thyroxine treatment. ATPase activity of cardiac myosin was stimulated as the Ca2+ concentration in the incubation medium increased. No difference was found in Ca2+-activation, salt sensitivity or ATPase activity of unreacted and sulphydrylmodified cardiac myosins from euthyroid or hyperthyroid groups. The results showed that in hyperthyroid rats, in contrast to some other species, the biochemical mechanism responsible for the enhancement of cardiac contractility is not an increased myosin ATPase.


Transferable drug resistance of Escherichia coli isolated from infant diarrhoea.

The resistance to several drugs was determined in 29 pathogenic strains of Escherichia coli (026, 055 and 0111) isolated from infant diarrhoea and 18 non-pathogenic E. coli strains isolated from the same individuals. Both pathogenic and nonpathogenic strains were resistant to at least 1 to 10 drugs, but only in four cases resistance patterns of the pathogenic strains were identical with those of non pathogenic ones. The majority of the strains were resistant to sulfonamide, tetracycline, ampicillin, carbenicillin, neomycin and kanamycin. The drug resistance (except the resistance to nalidixic acid and rifampicin) was associated with conjugative R-plasmids. Some of the tested strains carried two R-plasmids in one cell, being in hetero R-state.


Fungal nucleic acids as interferon inducers.

Nucleic acids isolated from the fungi Aspergillus niger x11, Piptoporus betulinus and Ganoderma applanatum reduced the number of vaccinia virus plaques in chick embryo fibroblast (CEF) tissue culture and when administered intravenously to white mice protected them against lethal infection with tick borne encephalitis virus strain K5 (TBE). In CEF tissue culture the nucleic acids of the studied fungi were found to induce small but detectable amounts of a substance with the character of interferon. In vivo only ribonucleic acid from G. applanatum induced a substance showing interferon properties in the spleen of mice.


IgM antibodies in rubella infections: absorption with staphylococcal protein A of sera examined by immunofluorescence and haemagglutination inhibition.

A similar frequency of positive results for rubella IgM antibodies in the sera of rubella patients and persons contacting such patients was obtained by immunofluorescence after separation of IgM is sucrose gradient and after absorption of the sera with Staphylococcus aureus and immunoglobulin aggregates. Compared to the immunofluorescence method, the haemagglutination method for rubella IgM antibodies performed after absorption of the sera with S. aureus and reduction of immunoglobulins M with 2-mercaptoethanol was less sensitive for serum samples taken in the acute phase of the disease and 5 weeks after the appearance of rubella symptoms.


Removal of nitrogen from industrial wastewaters by three-step nitrification and denitrification.

The removal of nitrogen from industrial wastewaters carrying about 1,000 mg NH4-N and urea-N/l was investigated on a laboratory scale. The use of a three-step nitryfying activated sludge with adjustment of pH from step to step resulted in 99% oxidation of both forms of nitrogen to nitrites. The efficiency of nitrification was 18 mg N/l/h. Total time of wastewater aeration depended  on nitrogen concentration and was 33-54 hours. Complete dentrification of NO2-N was obtained in packed-bed reactor with the use of acetic acid as a carbon source. Efficiency of denitrification was 361 mg N/l/h.


Rhizosphere mycoflora of healthy and yellow vein mosaic virus infected okra (Abelmoschus esculentus) plants.

Investigations on the rhizosphere mycoflora of healthy and virus (YVMV) infected okra plants showed a higher fungal population in the rhizosphere of healthy plants at preflowering and post-flowering stages than in that of diseased ones. Maximum population was observed during flowering both in healthy and diseased plant rhizosphere as well as in non-rhizosphere soil. However, virus infected plants showed a higher population at the flowering stage than healthy ones. The quantitative differences in the rhizosphere of healthy and diseased plants during flowering seem to be due to a change in C/N ratio and amino acids. The drastic reduction in diseased plant rhizospheres during the post-flowering stage may be due to either change in C/N ratio unfavourable to mycoflora or production of some toxic substances inhibiting multiplication of the mycoflora.


Occurrence and distribution of rhizobiophages in Indian soils.

Soil samples from 30 fields in neighbouring districts of Varanasi were screened for rhizobiophages on 60 Rhizobium strains. Plaques were observed on five strains: P1, P5, SU391 (R. leguminosarum), CB756 and 32H1 (Rhizobium sp.). Rhizobiophages infective on one or more of the five strains were present in all fields. There seems to be no correlation between presence of phage and the standing crop or the pH (7.1-8.2) of the soil. Eight distinct rhizobiophages have been isolated and characterized for host specificity, plaque morphology and maximum titer in broth.


Glucose and phosphorus utilization by two dermatophytes.

Trichophyton mentagrophytes and Aspergillus fumigatus dissimilated glucose through EMP pathway. Oxygen and inorganic phosphorus were found to control the rate and extent of glucose metabolism.


Macrophage-lymphocyte interaction and genetic control of immune responsiveness.

We have reviewed briefly some of the diverse functions of macrophages in the immune response. Clearly, this population of cells interact physically with lymphoid cells, are required for activation of T cells, and process various protein antigens. Finally, we have studied the immune response to insulin in order to unify these previous data in such a way to demonstrate the active role of macrophages in the regulation of the immune response. The function of the Ir gene in the guinea pigs appears to be an intramolecular selection of discrete regions within the antigen for recognition by the T cell. The data presented suggest that this function operates at the level of the macrophage.


A novel biological function of macrophages associated with antigen discrimination properties.

The ability of fresh or cultured subpopulation of adherent peritoneal exudate cells to perform discrete biological functions commonly associated with macrophages, namely antibody dependent cytotoxicity (ADCC) and phagocytosis, has been compared with the ability of the same cells to present TNP coupled to carbohydrate or protein carriers for a subsequent antibody response, or semi-allogeneic cells for T cell cytotoxic responses, in tissue culture. Data are presented to show that ADCC reactivity rapidly disappears on cultures of peritoneal cells, though phagocytic activity is more persistent throughout the 14 days of culture. When ADCC activity reappeared it was no longer restricted to those fractions active initially. Antigen handling of different TNP-protein conjugates was pronounced throughout the culture period and was not easily explained in terms of the ADCC/phagocytic activity of the macrophage subpopulation concerned. In contrast, however, antigen handling of carbohydrate antigens for presentation to T or B lymphocytes by the different macrophage populations varied during the culture period in fashions which were in some degree correlated with the phagocytic activity of the various cells but were also a function of the antigen under consideration. These data are interpreted in terms of a novel role for the macrophage in discrimination of carbohydrate antigens for presentation to lymphocytes.


Cardiac specific antigen and antibody in immunopathogenesis of cardiac disease.

Antibody to human cardiac extracts was detected in the sera of many patients undergoing open heart surgery. A passive hemagglutination assay using cardiac extracts as antigen coated onto erythrocytes showed the development of cardiac antibody in the patients, with peak titers usually one to three weeks after the surgery. Radioimmunoassay revealed the presence of cardiac specific myoglobin antigen antigen in the sera of many patients, with peak levels 1-3 days after the procedure. A correlation was observed between development of circulating serum antibody and appearance of cardiac myoglobin antigen. These results support the view that immunologic disease related to release of cardiac antigen in patients undergoing heart surgery may be a factor in post-cardiotomy disease.


The effects of metiamide and H1 receptor blocking agents on anaphylactic response in guinea-pigs.

The effects of metiamide and of four H1 receptor blocking agents (mepyramine, promethazine, clemastine and ketotifene) on anaphylactic reaction were studied in the guinea-pig. The H1 blockers conferred partial protection which shows that with the experimental protocol utilized (challenge injection with high doses of antigen), histamine plays a lesser role than other mediators released or synthesized. Metiamide (30.0 mg/kg i.v.) noticeably enhanced the increase in pulmonary resistance observed during anaphylactic reaction and reduced the protective effect of the H1 antagonists on this parameter and on histamine release. These effects might be explained by an inhibition - at least partial - of the negative feed-back mechanism through which histamine controls its own release, or by a specific action of metiamide in high doses. The transient tachycardia initially observed in anaphylactic shock is partly related to stimulation of cardiac H2 receptors by the histamine released, since it is suppressed by metiamide.


Adjuvant-induced inflammatory disease in the rat: plasma levels of peptide hydrolases and protease inhibitors reflect disease activity.

Both during the primary (localized) inflammation and the development of the secondary (generalized) inflammation in adjuvant-treated rats, the plasma level of functional alpha-macroglobulins increases while proteases (measured as peptide hydrolases) sharply decrease. The decreased peptide hydrolase levels during episodes when protease 'spillage' into the bloodstream is elevated, suggests a more rapid clearance of alpha-macroglobulin-protease complexes associated with inflammation.


Laboratory characteristics of four strains of goatpox virus.

Significant differences were found between four strains of goatpox virus isolated from goats in Egypt, Iraq, Iran and the U.S.S.R., namely in their growth behaviour in embryonated eggs, ceiling temperature, plaque characteristics in cell culture, heat resistance and in the degree of inhibition of their replication by actinomycin D and bromodeoxyuridine. All the strains studied had morphologically similar virions and induced type A V- inclusions. They were uncapable to produce haemagglutinins in infected cell cultures. Antigenically, the strains were indistinguishable.


Influence of human leukocyte interferon on IgG on the replication of herpes simplex virus in nervous tissue in vitro.

Herpes simplex virus type 1 (HSV-1) replicated productively in rabbit and guinea pig ganglia and nerve organ cultures when inoculated in high titres. Treatment with IgG 20 hr before and 48 hr after infection produced a delay of 4 to 7 days in the recovery of HSV-1 by the method of co-cultivation. The same result was obtained when IgG was combined with human leukocyte interferon. There was no difference in the period up to HSV recovery between the groups treated with interferon alone and the HSV control. Morphological evidence by light and electron microscopy of viral productive infection was obtained in all the cell types of nervous tissues infected in vitro.


The effect of native and sonicated double-stranded polyribonucleotides on the course of spontaneous autoimmune disease in NZB and NZB/Swiss F1 mice.

The administration of sonicated fractions of f2 phage polyribonucleotides caused an increased weight loss and deterioration of the clinical state in female NZB mice. Discontinuance of the treatment resulted in an improvement of both the clinical state and the genetically determined autoimmune disorders of these mice. Some potential explanations of this effect are discussed.


Serological relationships between Robinia mosiac, clover blotch and peanut stunt viruses.

Based on the results of agar gel double immunodiffusion tests with broad spectrum rabbit antisera and narrow spectrum mouse immune ascitic fluids and formalin-fixed purified viruses, a close relationship was established between 3 members of the Cucumovirus group namely Robinia mosaic virus (RoMV), clover blotch virus (CBV) and peanut stunt virus (PSV). The relationship of these 3 viruses to cucumber mosiac virus proved to be more distant. The results also suggested that RoMV and CBV could represent distinct strains of PSV.


A further member of the Yucaipa group isolated from the common wren (Troglodytes troglodytes).

In 1977, 477 small birds of 52 species were virologically examined. Five cloacal swabs from Troglodytes troglodytes yielded two paramyxovirus strains which were found antigenically related to, but not identical with Ch/Yucaipa/California/56 virus according to three serological tests. The ecological significance of these findings and the possibility of paramyxovirus classification into four types are discussed.


The plasma kinetics of hydroxyethyl starch 350/0.60: a potential new adjunct for centrifugal leucapheresis.

A modified species of hydroxyethyl starch (HES 350/0.60) possessing a Mw- of 350,000 daltons combined with a molar hydroxyethyl group substitution (MS) of 0.60 (60 hydroxyethyl groups.100 glucose residues) was clinically assessed in seven normal subjects to determine the influence of these chemical modifications on intravascular clearance kinetics concomitantly with effects on the suspension stability (ESR) of blood. Following a standardised intravenous dose (30 gm.m2 BSA), the concentration of HES 350/0.60 in serum fell to half its peak value in 11.8 +/- 1.3 (SD) hours, while the ESR remained elevated for up to 12 hours post-injection. By adopting a Mw- of 350,000 daltons, the critical molecular weight (Cmw) of this colloid was surpassed, while the critical concentration (Cc), below which the suspension stability of blood is not affected, was shown to range between 0.3 and 0.5 gm.dl-1. In comparison to the present species of HES (Mw- 450,000 daltons, MS: 0.70) utilised as a sedimenting agent duirng centrifugal leucapheresis, HES 350,000/0.60 appears to affect the ESR in a similar manner, but is removed from the intravascular space approximately twice as rapidly. This more rapid clearance should be useful in avoiding cumulative build-up of HES in blood concomitant with reducing the total amount of intravascular H2O bound to this colloid, in normal and CML donors undergoing multiple cell collection procedures.


Transient myeloid metaplasia associated with an unusual hemoglobin in a newborn infant.

A newborn infant presented with hepatosplenomegaly, rash, anemia, and leukocytosis at one day of age and manifested characteristic myeloid metaplasia by one mouth of life. Vitamin B12 and leukocyte alkaline phosphatase were elevated and platelet aggregation was impaired. Myelofibrosis was not present and neutrophil function was preserved. An unidentified high isoelectric point hemoglobin with unusual chromatographic and electrophoretic behaviors was found to comprise 12% of the total hemoglobin. The myeloid metaplasia and mutant hemoglobin disappeared over the subsequent months without biochemical or clinical residual. The available evidence was consistent with the mutant hemoglobin representing either a gamma chain or clonal embryonic chain variant. The inability to clarify prognostic factors in these unusual myeloproliferative syndromes suggests caution in the initiation of cytotoxic therapy.


Detection and quantitation of Dermatophagoides antigens in house dust by immunochemical techniques.

Rabbit antibodies against D. pteronyssinus whole culture extract were rendered specific to mite antigens by immunoabsorption with extract of human dander. Using the absorbed antibodies, an extract of a mite culture gave rise to 15 peaks in crossed immunoelectrophoresis and to a titer of 512 in counter-immunoelectrophoresis. When applied to dust samples from the homes of allergic patients, counterimmunoelectrophoresis gave reproducible results within an error of 1 titer-step. Mite antigens were detected in 64% of 105 dust samples investigated. The titer correlated well (P less than 0.005) with the concentration of Dermatophagoides bodies, found in 13 mattress dust samples by microscopy. Advantages and limitations of immunochemical quantitations of house dust mites in dust samples is discussed in relation to investigations by microscopy.


Cyclic AMP in hypersensitivity reactions.

Phytohemagglutinin (PHA) was capable of inducing non-cytotoxic histamine release from human leucocytes. In the presence of deuterium oxide (D2O), PHA caused significantly greater histamine release. Dibutyryl cyclic AMP (d-cAMP) could enhance the histamine release in the presence of D2O although it was an inhibitor of the release if used alone. However, a beta agonist, isoproterenol, which increases intracellular level of cAMP was inhibitory with or without D2O. These data ask the question about dual effect of cAMP and suggest the possibility of different polls of cAMP in the target cells.


Frontiers of quantitative cytochemistry: a review of recent developments and potentials.

This article attempts to describe the current state of quantitative cytochemical methods and to point out areas where, in the author's opinion, the greatest progress is either being made or can be expected. Nearly 300 references to the current literature are cited.


Comparison of two Papanicolaou staining procedures for automated prescreening.

A "regressive" (procedure I) and a "progressive" (procedure II) modification of the Papanicolaou stain were tested for their suitability for automated screening procedures. Intermediate squamous cells and carcinoma cells were scanned with a high-resolution system, and digitized data were statistically analyzed after feature extraction. The regressive modification accentuates the textural features of intermediate squamous cell nuclei as well as the contrast ratio of nucleus and cytoplasm. The progressive modification accentuates some criteria of malignancy, such as polychromasia, hyperchromasia and shifting of nuclear to cytoplasmic ratio. Grouping of data according to patients indicates that the differences found are due to the staining procedures and not to individual variability of parameters in different patients.


The monodisperse cervical smear: quantitative analysis of cell dispersion and loss with enzymatic and chemical agents.

The proper utilization of flow-through instruments for the automatic detection of malignant cells in human cervical specimens requires that the cells be in the form of a monodisperse suspension. Information regarding the degree of cell dispersion and of cell loss is therefore of critical importance in the evaluation of any procedure used to render cervical specimens monodispe automatic detection of malignant cells in human cervical specimens requires that the cells be in the form of a monodisperse suspension. Information regarding the degree of cell dispersion and of cell loss is therefore of critical importance in the evaluation of any procedure used to render cervical specimens monodispe automatic detection of malignant cells in human cervical specimens requires that the cells be in the form of a monodisperse suspension. Information regarding the degree of cell dispersion and of cell loss is therefore of critical importance in the evaluation of any procedure used to render cervical specimens monodisperse. We have devised a simple method for the simultaneous assessment of these two parameters using smears prepared with the Perkin-Elmer Uni-Smear Spinner. Our quantitative evaluation indicates that none of the 15 enzymatic and chemical agents tested to disperse cervical specimens produced an adequate monodispersed cell suspension without unacceptable cell loss. Electron microscopic evidence is used to illustrate the deleterious effects of some of the agents employed.


A poststaining fixation technique for acridine orange: quantitative aspects.

A poststaining fixation technique was developed for acridine orange staining of cytologic specimens. A quantitative evaluation was performed to assess the suitability of this technique for routine use in analytic cytology and its potential for future use in automated screening equipment. Three major advantages of the poststaining fixation technique over conventional acridine orange staining methods were documented: (1) stained cells may be stored for several months without alteration of cellular morphology or fluorescence characteristics; (2) the protocol minimizes diffusion of stain out of cells; and (3) poststaining fixation reduces dependence of nuclear or total cell fluorescence on cell distribution or concentration on a slide or in flow.


Chronic metabolic effects of ammonia in mouse brain.

Chronic ammonia toxicity in experimental mice was induced by exposing them for 2 and 5 days to 5 % (v/v) ammonia solution. The enzymes concerned with glutamate metabolism (aspartate-, alanine- and tyrosine aminotransferases, glutamate dehydrogenase and glutamine synthetase) and (Na+ + K+)-ATPase were estimated in the three regions of brain (cerebellum, cerebral cortex and brain stem) and in liver. Glutamate, aspartate, alanine, glutamine and GABA, RNA and protein were also estimated in the three regions of brain and liver. A significant rise in the activity of (Na+ + K+)-ATPase in all the three regions of brain along with a fall in the activity of alanine aminotransferase was noticed. Changes in the activities of other enzymes were also observed. A significant increase in alanine and a decrease in glutamic acid was observed while no change was observed in the content of other amino acids belonging to the glutamate family. As a result of this, changes in the ratios of glutamate/glutamine and glutamate + aspartate/GABA was observed. The results indicated that the brain was in a state of more depression and less of excitation. Under these conditions the liver tissue was showing a profound rise in the activity of the enzymes of glutamate metabolism. The results are further discussed.


An alpha-adrenotropic study of the normal and diabetic rabbit kidney.

Rabbit kidneys from normal and alloxan-treated animals were isolated and perfused at 30 degrees C, with Krebs-Henseleit solution. Norepinephrine (NOR), 1 microgram/min, promoted an increase in perfusion pressure which was blocked by phentolamine. In diabetic kidneys NOR induced a sluggish increase in perfusion pressure and resistance, showing a decrease in sensitivity of the adrenergic receptors to the drug. Propranolol, a beta-blocker, was able to elicit an alpha adrenergic blockade in diabetic kidneys. These facts demonstrate an adrenergic receptor defect in diabetic animals, which was shown just three weeks after alloxan treatment.


[Effects of precursors and metabolites of catecholamines on motricity of isolated rat duodenum. Particular reference to dopamine].

Studying the effects, on the isolated rat duodenum motricity, of ten compounds precursors or metabolites of catecholamines, the following results were obtained: The direct metabolites of epinephrine and norepinephrine (metanephrine, normetanephrine), are either ineffective at concentrations below 5 X 10(-6) M, or weakly inhibitory at higher concentrations. Such inhibitory effects are prevented by alpha- and beta-blockers. 3-methoxy, 4-hydroxyphenylglycol and vanylmandelic acid have no significant effect. The catecholamine precursor, dopamine, the related compounds DOPA, 3 methoxytyramine, and to a lesser extent, 3-O methyl DOPA and homovanillic acid, have excito-motor effects at concentrations ranging mainly from 10(-7) M to 10(-5) M. At higher concentrations, the same compounds frequently exhibit inhibitory effects. The excito-motor effects might be due to a serotoninergic mechanism, since they are suppressed by the serotoninergic blocking agents methysergide and cyproheptadine. Furthermore, in the case of DOPA, we were able to establish a relationship between the excito-motor effects and duodenal serotonin stores. As for the inhibitory effects, they may be prevented by using alpha and beta blocking agents. Dihydroxyphenylacetic acid has no effect on the isolated rat duodenum motricity. The fact that dopamine and related compounds may have excitomotor effects at some concentrations, correlated with some physiopathological observations in man and animal allows some considerations about the eventual role of dopamine on intestinal motricity.


Determination of metabolite compartition in hepatic cells by varying the redox state in vivo.

In the present study, metabolite (lactate, pyruvate, glycerol 3-phosphate, dihydroxyacetone phosphate) concentrations were measured in various redox states. The mathematical relations between metabolite concentrations in various redox states were expressed algebraically and studied. Models which provided separate lactate/pyruvate (L/P) and glycerol 3-phosphate/dihydroxyacetone phosphate (G/D) spaces correspond to the experimental results in the case of "reductants" (e.g. ethanol, acetaldehyde, dihydroxyacetone and acetate) or of "oxidizing agents" (e.g. pyruvate) of the cytosolic NAD-NADH. Crotonate injection caused an oxidation of cytosolic redox couples, but no separation of the lactate/pyruvate space from the glycerol 3-phosphate/dihydroxyacetone phosphate space may necessarily be inferred. Furthermore, the following statements could be made in both first cases: (i.e. of "reductants" and "oxidizing agents"): (a) Redox couples in L/P space and in G/D space (together L/P-G/D system) are in equilibrium; (b) Redox-equivalent transport from the L/P space to the G/D space is not subject to any velocity-limiting mechanism; (c) Substrates which transports redox-equivalents into and from the L/P-G/D system reach concentrations to values, which are in a linear relation to each other in this system; (d) It is possible that these substrates are regenerated in another system which is also in equilibrium and subject to statement c.


Cytochalasin E-induced oxidative metabolism in polymorphonuclear leukocytes.

The extent of the cyanide-resistent oxidative burst of polymorpho-nuclear leukocytes after stimulation with cytochalasin E was shown to depend markedly on the osmolarity of the cell-suspension medium. With granulocyte concentrations up to 2 X 10(6) cells/ml, optimal oxygen consumption and releases of H2O2 and superoxide anions were reached at 180 mOsmol and 2 X 10(-5) M cytochalasin E. After removal of unbound activator, the cellular oxidative activity remained unaltered and continued to depend on the used osmotic conditions. It is proposed that binding of cytochalasin to the plasma membrane induces an irreversible activation of the oxidative system, whereas the resulting metabolic activity depends on conformational changes in the plasma membrane.


Effect of beta-blockers on sodium and potassium content of human erythrocytes.

Recently, hyperpotassemia was reported in patients treated with propranolol, but the mechanism has not yet been delineated. We have investigated the effects of various beta-adrenergic blockers and of D-propranolol on the Na+ and K+ distribution intra- and extracellularly in human erythrocytes. K+ loss and Na+ gain by cells was demonstrated at drug concentrations of 10(-4) M or greater. D-Propranolol was more effective than L-propranolol, whereas pindolol was ineffective. Practolol increased Na+ content but did not influence K+. The results suggest that electrolyte redistribution across cell membranes is not a likely explanation for hyperpotassemia in patients treated with propranolol, or for the local anaesthetic effect of this drug.


The influence of sex, of castration in new-born males and of androgen treatment in new-born females on the mouse-killing behaviour of the rat.

The results we have got on the mouse-killing behaviour of the rat point out the remarkable percentage difference between control males and control females, between integral males and males castrated at birth, and between normal females and females under the effects of androgens. Such results make us believe that the appearance of the killer behaviour is affected by the presence of testosterone in the first days of life of the rat.


A research on the action of testosterone propionate and of ciproterone acetate on the mouse-killing behaviour of the adult rat.

Data demonstrate exactly that testosterone favour above all the killing behaviour of castrate males at birth in comparison with control males while ciproterone acetate turns out to be more effective in diminishing the killing behaviour of males having integral testicle and of androgenized females at birth in comparison with castrated males. The results point out that the killing behaviour of rats is affected by the presence of testosterone in the first days of life, and in the grown-up animal.


Effect of evisceration on the disposal of [14C] palmitate in the rat.

The utilization in vivo of [1-(14)C] palmitate was studied in hepatectomized-nephrectomized rats and their sham-operated controls. After i.v. injection of the tracer, the [14C] lipids in plasma disappeared more slowly in eviscerated animals than in their controls. More label reappeared in plasma as esterified fatty acids in the latter group. At 30 min after the tracer, the amount of label found in the lipidic fraction of carcass and heart was much greater in eviscerated animals than in their controls although the percentile distribution of labelled lipidic fractions remained stable, a considerable proportion being present in the esterified fatty acid form. On the basis of these findings, the rapid increase in the plasma levels of FFA in eviscerated animals must be the result of augmented lipolytic activity more than reduced utilization of these metabolites.


Thyroid activity in a hypometabolic primate, the owl monkey (Aotus trivirgatus).

Serum levels of triiodothyronine (T3) and tetraiodothyronine (T4) were significantly lower in owl monkeys than in long-tailed macaques. These observations were considered to be consistent with the lower metabolic rate of the owl monkey. However, the absence of a significant difference in the levels of thyroid stimulating hormone (TSH) between the two species suggested a lower thyroid sensitivity to TSH in the owl monkeys. There was an inverse relation between levels of T3 and TSH in the owl monkeys at night and during the day.


[Chemoreflexive drive of ventilation and noradrenaline stimulus in man].

O2 chemoreflex drive of ventilation was studied before and after an intravenous infusion of L-norepinephrine (9 microgram/min), inducing a plasmatic hormone concentration similar to that obtained during submaximal exercise. The ventilation increasing rapidly at the beginning of the infusion was stabilized after 30 min : the ventilation was two times the reference value. The O2 chemoreflex drive of ventilation increased during norepinephrine infusion. When man was transiently switched from hypoxia to pure O2 (O2 test) the maximal fall of ventilation was two times the reference response. This increase in the chemoreflex drive, although the physico-chemical blood state was unchanged, may be explained by a norepinephrine chemoreceptor sensitization. Such a mechanism could partly explain the increase of O2 chemoreflex drive observed during muscular exercise.


Thiamine intestinal transport and phosphorylation : a study in vitro of potential inhibitors of small intestinal thiamine-pyrophosphokinase using a crude enzymatic preparation.

Using as enzymatic source the cytoplasmatic fraction of enterocytes isolated from the rat small intestine, thiamine-pyrophosphokinase activity was studied with a radiometric method using [thiazole-2-(14)C] thiamine. The Km value for thiamine was 2.14 X 10(-6) M and V 0.87 nmol of thiamine pyrophosphate mg-1 protein h-1. Eleven thiamine structural analogs and derivatives were assayed for their inhibitory action on the small intestine thiamine-pyrophosphokinase activity. Their Ki values were : pyrithiamine, 2.25 X 10(-6) M; thiamine monophosphate, 4 X 10(-6) M; 2'-ethylthiamine, 8 X 10(-6) M; 2'-butylthiamine, 6 X 10(-6) M; chloroethylthiamine and dimethalium, 1.5 X 10(-5) M; amprolium, 1.8 X 10(-4) M; L-582571, 1.65 X 10(-4) M; oxythiamine, 4.2 X 10(-3) M. Of the miscellaneous compounds tested (toxopyrimidine, Na-pyrophosphate, choline, L-phenylalanine, ethyl-urethane and 5-fluorouracil), none had any inhibitory action on intestinal thiamine-pyrophosphokinase activity, even if used at concentrations hundred times higher than that of labelled thiamine.


Protein nutrition of Tenebrio molitor L. XX. Growth response of larvae to graded levels of amino acids.

Larvae of Tenebrio molitor L;, Gembloux strain, race F, having an average initial weight of 10 mg were reared for 4 weeks at 27 +/- 0.25 degrees C and 65 +/- 5% relative humidity on diets containing an amino-acid mixture resembling the composition of larval tissues; Each of 14 amino acids was tested individually at levels of 0, 10, 20, 50, 100 and 200% of the amount found in larval tissues, while the concentration of other amino acids remained constant. Diets were maintained isonitrogenous at 1.6% nitrogen with supplementary glutamic acid, glycine, serine and tyrosine; Maximum fresh weight gains and dry weights were achieved by larvae fed reference levels of all essential amino acids except phenylalanine, threonine and tryptophan. Maximum growth was achieved by larvae fed 50% of the phenylalanine level found in larval tissues, Threonine and tryptophan were the limiting amino acids in this study and are probably required in the diet in excess of twice the concentration occuring in larval tissues, Probable ranges for quantitative amino-acid requirements of T. molitor were determined and suggestions were made for improving the nutritional adequacy of the amino-acid mixture.


Antigenic homology of feline and human beta-hexosaminidase.

The immunological characteristics of feline beta-hexosaminidase (beta-D-N-acetylglucosaminidase, EC 3.2.1.30) isoenzymes, Hex A and Hex B, were studied. Immunization of rabbits and goats with either cat Hex A or Hex B produced antibodies which reacted with a common antigenic marker shared by both Hex A and Hex B. With properly absorbed antisera, a unique antigenic marker was demonstrated on cat Hex A, but not on Hex B. This antigenic profile is comparable to that of the human beta-hexosaminidase isozymes, in which both Hex A and Hex B share the antigenic determinant, beta, while only Hex A possesses the antigenic determinant, alpha. Ho cross-reactivity between the two species could be demonstrated using goat or rabbit antisera to either feline of human beta-hexosaminidase. These immunological data validate feline Gm2 gangliosidosis as a model for human Gm2 gangliosidosis type II, and facilitate the investigation of enzyme replacement therapy.


Purification and properties of rat alpha 2 acute-phase macroglobulin.

Alpha 2 acute-phase macroglobulin was isolated from plasma of turpentine-injected rats. In the method conditions known to damage the biological activities of alpha 2 macroglobulin are avoided. The procedure successively involves: rivanol precipitation, concanavalin A-Sepharose chromatography and ion-exchange chromatography on DEAE-cellulose. Proteolytic activities were minimized throughout the purification. Thus alpha 2 macroglobulin was obtained in a 20% yield and was pure by biochemical and immunological criteria. Its molecular weight appeared to be 760 000 and it consisted of four subunits (Mr 190 000). The protein has an A1cm 1% = 8.8 and an isoelectric point = 4.8. The amino acid and carbohydrate compositions were determined. Our preparations bound 1 molecule of trypsin or 1 molecule of plasmin/molecule of alpha 2 macroglobulin. Kinetic parameters for alpha 2 macroglobulin-bound trypsin and plasmin were determined and compared with those of free trypsin and plasmin using butoxycarbonyl-L-valylglycyl-L-arginine-2-naphthylamide and benzoyl-L-arginine ethylester as substrates.


Mechanism of binding of mouse interferon to controlled pore glass.

Many proteins bind to controlled pore glass; they are either acid elutable or alkali elutable. Mouse interferon is an acid-elutable protein. Since poly(L-lysine) and, to some extent, poly(L-arginine) are also eluted from controlled pore glass under acidic conditions, one may postulate that mouse interferon binds to controlled pore glass via some of the protein's epsilon-amino groups (of lysine) and/or guanidinium groups (of arginine) and the beads' silanol (hydroxyl groups). The necessity of lysine in the binding of interferon to controlled pore glass is further substantiated by the fact that citraconylated interferon does not bind to controlled pore glass. A requirement for Lewis acid-base interaction between the beads' B2O3 groups and the amide groups of arginine is unlikely in view of the results obtained with the alternative system, ZrOH, which, being devoid of B2O3, did bind interferon. Since a substantial amount of interferon could be eluted from controlled pore glass with ethylene glycol and high salt, one may assume that some hydrophobicity is involved in the binding of interferon to controlled pore glass.


Internalization of the amino terminal 1-78 sequence of the gamma chain of native fibrinogen and exposure associated with catabolism and plasmin cleavage.

Specific antibodies of the gamma 1--78 peptide of human fibrinogen were employed in binding assays and in equilibrium competitive inhibition assays to analyze the expression of gamma 1--78 antigenic determinants as an indication of the relative exposure of the gamma 1--78 sequence in the E domain of fibrinogen, high solubility fibrinogen subfractions I-8 and I-9, and plasmic cleavage fragments of fibrinogen and fibrin. A very limited exposure of gamma 1--78 sequences was found to occur concomitant with proteolytic deletions of the major carboxyterminal segment of the A alpha chains in fgI-8, fgI-9. Exposure of gamma 1--78 is not influenced by further proteolysis to fg-X which is associated with B beta 1--43 deletion. Further proteolysis to fg-Y, which is associated with deletion of beta 43--53 and of one of the D domains, is associated with additional exposure of gamma 1--78. This is not significantly influenced by further proteolysis to fg-E with deletion of the second D domain, deletion of A alpha 1--19, and proteolysis at the carboxyterminal aspects of the E domain chains.


[Saliva secretion, concentration and secretion of human salivary amylase in the 1st year of life].

This work show the levels of the salivary secretion volume in 5 min., the salivary amylase concentration in U/ml and the salivary amylase secretion in U/kg/5 min. in 10 children at 4, 15, 30, 45, 60, 90, 120, 150, 180 days of life and at 9 and 12 months. All the children had the same nourishment orientation and showed a good weight and neuromotor development during the period of the observation. The datas were compared with datas of 18 children with 20-36 months of age and 18 healthy young man. It was standardized the technic and the time-table to collect the salivary secretion. It was observed that the salivary secretion is higher between 90 and 180 days of age preceding theeth eruption. At 12 months of life the levels of the amylase salivary concentration reach the levels of infants and adults. The salivary amylase secretion is high after the 60 days of life. Probably this is responsible by the good starch tolerance at this time.


Hydrophobic interaction of fluorescent probes with fetuin, ovine submaxillary mucin, and canine tracheal mucins.

The presence of hydrophobic sites in fetuin, ovine submaxillary mucin and two homogeneous canine tracheal mucins was established by fluorescence probe techniques. The interaction between the above-mentioned glycoproteins and two hydrophobic fluorescent compounds, sodium mansate and mansylphenylalanine, was accompanied by an enhancement in fluorescence and a shift of the fluorescence maxima to shorter wavelengths. The introduction of a phenylalanine residue to the mansyl group enhanced the binding affinity of the probe for the hydrophobic sites of these glycoproteins as evidenced by lower values for the dissociation constants. The high molecular weight (581 600) tracheal mucin, which had the highest carbohydrate content (80%) of all the glycoproteins investigated, exhibited the highest fluorescence enhancement and the largest number of binding sites for these fluorescent probes.


[Effects of different concentrations of chloramphenicol on RNA synthesis in E. coli].

In the presence of 2 microgram/ml chloramphenicol the rate of total RNA synthesis increases 1,6-fold, while that of rRNA synthesis--2,5--3,0-fold. Other ribosomal antibiotics, e.g. oxytetracycline, neomycin and puromycin, as well as chloramphenicol stimulate the synthesis of RNA while not causing a complete inhibition of the protein synthesis. Intensive stimulation of the rate of rRNA synthesis with low concentrations of chloramphenicol may be due to a decrease of intracellular concentration of guanosine-5'-diphosphate, 3'-diphosphate, as well as to the maintenance of a sufficiently high level of protein synthesis under these conditions.


[Study of membrane proteins from Microccus lysodeikticus using immunochemical methods].

Using immunoelectrophoresis, the antigenicity of various protein fractions of the Micrococcus lysodeikticus membranes was evaluated. It was shown that both the peripheral and integral membrane proteins possess the antigenic determinants. The antiserum exhausted by the M. lysodeikticus mebranes loses its ability to interact with intergral proteins, which are not solubilized by Triton X-100. It was thus assumed that the integral proteins are exposed on the membrane surface constantly or periodically and that there exist no proteins which are completely and permanently incorporated into the lipid bilayer. The respiratory chain of the M. lysodeikticus membrane is inhibited by membrane immunoglobulins by 50%. This is probably due to the presence in the membrane antiserum of antibodies specific to the respiratory chain enzymes. Evidence for this assumption can be derived from the fact that partially purified cytochrome b556 forms a precipitation zone with the membrane antiserum and that the activity of membrane NADH-dehydrogenase is inhibited by a monoserum against NADH-dehydrogenase.


[Role of calcium in realization of nervous control during RNA synthesis in skeletal muscles].

The effects of Ca2+ on the RNA polymerase activity of the nuclei isolated from normal and denervated gastrocnemius muscles of the rabbit were studied. It was shown that 18 hrs after denervation the RNA synthesis in vitro, Ca2+ content and the Ca, Mg-ATPase activity of the nuclei are decreased. After addition of exogenous Ca2+ the incorporation of labelled UTP into the nuclei is stimulated in the denervated muscle and is inhibited in the control. Electrostimulation of the denervated muscle at the peripheral part of the sciatic nerve for 3 hrs increases both the RNA synthesis in the nuclei and the Ca2+ content, as well as the Ca, Mg-ATPase activity. Exogenous Ca2+ has an inhibitory effect on the nuclei of the stimulated muscle. The correlation established is indicative of participation of Ca2+ in the transmission of excitation in skeletal muscle sarcolemma to the processes occurring in nuclear structures.


Genetic control of aldehyde oxidase activity and cross-reacting-material in Drosophila melanogaster.

Four different genes are known to affect aldehyde oxidase activity (AO) in Drosophila melanogaster. Mutants at each of these loci eliminate AO activity and simultaneously eliminate detectable AO-crossing reacting material (AO-CRM) even though only one is the structural gene for AO (Aldoxn). The other three genes (cin1, lxd and mal) coordinately "control" the levels of activity of AO and two related enzymes, xanthine dehydrogenase (XDH) and pyridoxal oxidase (PO). Contrary to their effects on AO-CRM, neither of these three mutants eliminate XDH-CRM. A model of interaction of these enzymes and genes controlling their activities is discussed.


The effects of lao on aldehyde, oxidase activity and cross-reacting-material in Drosophila melanogaster.

In Drosophila melanogaster aldehyde oxidase occurs in at least two forms that can be separated electrophoretically. The mutant allele lao (low aldehyde oxidase activity) causes a deficiency of the major form of this enzyme. Immunoelectrophoretic analyses suggest that lao homozygotes produce aldehyde oxidase cross-reacting-material in nearly wild-type levels. Although aldehyde oxidase from the mutant stock is heat labile. properties such as Km and pH optima are not different from the normal enzyme.


Influence of post-extrasystolic potentiation on left ventricular function estimated by means of systolic time intervals.

Systolic time intervals were measured in 50 patients with frequent premature ventricular beats. The patients were divided into two groups: group I included those which showed in the beat that preceded an extrasystole a pre-ejection period/left ventricular ejection time (PEP/LVET) ratio greater than or equal to 0.43, and group II with PEP/LVET ratio greater than 0.44. Systolic time intervals recorded during post-extrasystolic potentiation were compared with those measured in the preextrasystolic complex. Also the measured intervals were tested against the hourly rate of premature beats obtained by electrocardiographic telemetric monitoring. The results confirmed the following results of previous reports: a) ventricular premature beats are followed by sinus-potentiated contractions inversely related to the coupling interval and dependent on adequate compensatory pauses; b) potentiated contractions are greater in patients with ventricular dysfunction. No relationship was found between left ventricular performance and the rate of premature beats. It is concluded that the effect of an antiarrhythmic intervention on the left ventricular function might be adequately evaluated by means of systolic time intervals and provoked post-extrasystolic potentiation, with the advantage of using totally non-invasive procedures.


Lack of leukocyte migration inhibition by hepatitis B surface antigen in hepatocellular carcinoma patients.

Soluble part of hepatocellular carcinoma (HCC) tissue extracts with or without hepatitis B surface antigen (HBsAg) was tested against leukocytes of 13 histologically confirmed HCC patients. Inhibition of leukocyte migration was observed in 9 out of 13 cases when tested by soluble HCC extract containing HBsAg, while inhibition of lukocyte migration was observed in 8 out of 13 cases when tested by solublp greater than 0.05, by Fisher's exact test). In the meantime, soluble HCC extract with or without HBsAg did not significantly cause inhibition of leukocyte migration in 12 non-HCC patients. Therefore, it is concluded that inhibition of leukocyte migration in HCC patients is caused by the tumor-associated antigen, not caused by HBsAg.


[Faecal enzyme diagnostic (author's transl)].

The hitherto existing results of determinations of enzymatic activities in stool are presented in this review. The chymotrypsin activity is diminished in advanced exocrine pancreatic insufficiency. The faecal alpha-amylase activity has up to now no significance in the diagnosis of pancreatic diseases. Up to five amylolytic enzyme activities are detectable. The alkaline phosphatase is mostly of intestinal origin. Up to 4 enzyme bands can be exhibited with the disc electrophoresis in polyacrylamide gel. Lysozyme and N-Acetyl-beta-D-glycosaminidase can also be detected in stool.


[Animal experiments on the induction of humoral antibodies by repeated implantations of dental gelatin and dried fibrin foam].

An attempt was made to demonstrate humoral antibodies against porcine and bovine protein in the sera of guinea pigs following repeated subcutaneous implantation of Gelastypt, Gelfoam, and Fibrospum. Precipitated antibodies against porcine protein could not be demonstrated with the agar gel double diffusion test in the sera of those animals in which gelatin substances had been implanted (starting protein: porcine collagen). Precipitated antibodies against bovine protein were demonstrable in the sera of all animals treated with Fibrospum (starting protein: bovine fibrin). The active hemagglutination test with porcine erythrocytes using sera taken from animals treated with gelatin substances showed a positive to high antibody titer in three cases. The same test carried out with bovine erythrocytes and sera from animals treated with Fibrospum showed positive to high antibody titer for all animals tested.


[Characteristics of human dental pulp cells in cell cultures].

The generation time and the division rate were measured with routine methods of cell culturing. The conditions contributing to the stagnation of growth were examined; the synchrony of the growth process was investigated. The biochemical composition of the cells was assessed via colorimetry and then compared with other cell systems. The cell culture examined may be used to test capping materials; only cells up through the eighth generation however should be utilized.


The effect of glutaurine on the development of amphibian larvae with inhibited RNA and protein synthesis.

1. Glutaurine (gamma-L-glutamyl-taurine) isolated from bovine parathyroid and later synthetized retards the metamorphosis of Rana arvalis. 2. From day 13 of treatment glutaurine moderates the metamorphosis inhibiting effect of antibiotics acting at the transcription and translation level.


The effects of aminotriazole (ATZ) on the thyroid gland and the development of the White Leghorn chick.

White Leghorn chicks, injected i.p. (days 3 through 40) with 0.5 g/kg or 1.0 g/kg mean body weight of aminotriazole (ATZ), gained less weight than the control animals. In the 1.0 g/kg ATZ group in comparison to the 0.5 g/kg animals, the responses were of greater magnitude and for longer durations of time. In both experimental groups, the thyroid/body weight ratios increased and were greater than those of the controls after day 10. In the higher dosage group, these ratios increased markedly, whereas with the lower dosage group, the ratios stabilized at a lower value. Histological alterations of the thyroid included hypertrophy, hyperplasia and hyperemia. To study the effect of cessation of drug treatment, groups of animals received the same dosages of ATZ through day 20. At the termination of the experiment (day 41), this resulted in a sharp increase in body weight, experimentals weighing as much as or more than the controls. Regarding the decrease in thyroid/body weight ratios, experimentals never attained the low control values. However, upon cessation of drug treatment, a return towards normal thyroid histology was noted. The gross body weight, the thyroid/body weight ratios, the histological analyses of the thyroidal components and the triiodothyronine and thyroxine determinations in the blood serum demonstrate that ATZ is a potent goitrogenic agent which produces a state of functional hypothyroidism.


Effect of glucocorticoid hormone on the content and synthesis of nucleic acids in cartilage of growing mice.

Glucocorticoid hormones are known to exert distinct inhibitory effects upon skeletal growth. This study examined the influence of triamcinolone hexacetonide, a long-acting synthetic analogue of cortisol, on nucleic acid synthesis in condylar cartilage of neonatal mice. It became evident that following a single injection of the hormone the DNA, RNA and protein contents were significantly reduced. The hormone inhibited both the uptake and incorporation of 3H-thymidine and 3H-uridine into DNA and RNA. Significant changes became apparent by 24 hours and persisted for 72 hours after administering the hormone. Thereafter full recovery was noted. Correlative relationship was noted between the inhibitory effects on the uptake and the subsequent incorporation of the above precursors into their respective nucleic acids. This study clearly indicates that corticosteroid hormones possess a significant inhibitory effect on the proliferative activity of neonatal chondrocytes and upon the latter's protein synthetic pathways, thereby affecting the normal process of endochondral bone growth.


On the pharmacology of venous smooth muscle from dog and man.

Changes in tension of spiral strips from canine and human veins induced by various drugs are compared with results from the literature on human veins. The order of potencies of alpha-adrenoceptor stimulating amines (adrenaline greater than noradrenaline greater than dopamine greater than isoprenaline) is similar in human and canine veins. Comparison of the affinities for alpha-adrenoceptors of pharmacologic drugs (thymoxamine, indoramine, clonidine, dihydroergotamine) suggest marked differences between the alpha-adrenoceptors in veins from man and dog. Venoconstriction mediating 5-HT receptors and a very small population of beta-adrenoceptors exist in both species. Human veins are always dilated by histamine, while canine femoral veins in vitro are relaxed by lower and contracted by higher histamine concentrations. Prostaglandin F2 alpha constricts both canine femoral and human hand veins. PGA2 and PGE2 increase the tension of canine and human veins in vitro but dilate human hand veins in situ. The order of potencies of ergot alkaloids in canine femoral veins is ergotamine greater than dihydroergovaline greater than dihydroergotamine = dihydroergostine greater than methysergide, whereas in human hand veins it is ergotamine = dihydroergovaline greater than methysergide greater than dihydroergotamine greater than dihydroergostine. In dogs the venoconstrictor effect of ergotamine is mediated by at least 3 mechanisms: stimulation of [1] alpha-adrenoceptors, [2] 5-HT receptors and [3] endogenous prostaglandin synthesis. Stimulation of alpha-adrenoceptors by dihydroergotamine and of 5-HT receptors by ergotamine was confirmed on human hand veins in situ. Prejunctional receptors at sympathetic nerve terminals are involved in the regulation of venous tone. Inhibitory alpha-adrenoceptors, dopamine and PGE2 receptors as well as facilitating beta-adrenoceptor existing at human vasoconstrictor nerves may be stimulated or blocked by pharmacologic drugs thereby modifying venous tone.


[Effect of ergoline derivatives on platelet and blood vessels].

The influence of dihydrogenated ergot alkaloids on vessels and blood platelets was studied in vitro and in vivo. At low concentrations they cause contraction of isolated vein strips via a stimulation of alpha-adrenoceptors, at higher concentrations they antagonize the action of noradrenaline on arteries and veins and inhibit the adrenaline-induced platelet aggregation in vitro. In volunteers, intravenous and subcutaneous administration of dihydroergotamine (Dihytamin) caused a decrease in blood volume of the capacitance vessels and inhibition of the adrenaline-potentiated platelet aggregation "ex vivo". The selective venoconstrictor and antiaggregating effects of dihydroergotamine are utilized in the postoperative prophylaxis of thrombosis.


Clinical pharmacology of phlebotropic drugs.

Methods used in clinical and clinical pharmacological assessments of phlebotropic drugs are outlined. The knowledge of the pharmacokinetics and metabolism of phlebotropic drugs in human organism is far from complete as yet. Of objective methods proving haemodynamic effects of phlebotropic drugs, the authors have the best experiences with the strain-gauge type occlusive plethysmography, which is capable of proving a reduction of the capillary filtration fraction even after a single administration of an active preparation to healthy volunteers. The control of the patient's subjective complaints by phlebotropic drugs is documented by statistically significant findings in double-blind clinical trials with a placebo.


[Prevention of postoperative phlebothrombosis by means of pneumatic massage of the extremities in animals].

Thromboembolism may be considered as the most serious complication in the postoperative period. Due to improvements in anaesthesia and postoperative supervision even those patients who are in a bad state of health may be operated upon. The frequency of postoperative phlebothrombosis, however, has increased in recent years. External pneumatic massage of the extremities will speed up the blood flow in the veins and increase the fibrinolytic activity in the vein wall. In animal experiments we could observe a significant decrease of the thrombus size in artificially produced thrombosis in the vena femoralis of dogs, if after thrombus induction the external pneumatic massage was applied.


Biochemical mechanism of platelet activation. Involvement of contractile proteins.

The present state of knowledge of the biochemical mechanism of platelet activation (adhesion, shape change, microspike formation, aggregation, release reaction, clot retraction) is presented under involvement of contractile proteins. The working hypothesis on the contractile mechanism of platelet activation is explained.


[Clinical aspects of control in therapy with platelet inhibitors].

46 patients with ischaemic heart disease were treated with Micristin (20-40 mg/kg) or a combination of Micristin and Propranolol (80-120 mg/die). The values of bleeding time, the platelet factor 4 in lysate of thrombocytes or in plasma of patients as well as the soluble fibrin monomer complexes were investigated. They showed no obvious correlation to the clinical findings.


[Clinical-experimental studies on the inhibition of thrombocyte function with acetylsalicylic acid and with indobufen].

The mode of action of two inhibitors of platelet function is reported. At low doses both substances inhibit platelet aggregation, at higher doses they inhibit the availability of platelet factor 3 and liberation of platelet factor 4. Acetylsalicylic acid, however, possesses a long-lasting effect produced by irreversible acetylation of platelet membrane. This was demonstrated in a series of clinico-experimental studies in which 14C-labelled ASA was used. Indobufen was found to cause transitory changes of platelet function. It is easily washed out from platelets and binds loosely to cytosol proteins of platelets, as shown in studies with 14-C-labelled indobufen. In contrast to ASA, it exerts an effect on platelets lasting for a few hours only.


[Prevention of reinfarction with acetylsalicylic acid].

A report is presented on the prospective trial for prophylaxis of myocardial reinfarction by means of acetylsalicyclic acid. For the first time it was organized on the basis of population. 1,340 patients were treated with acetylsalicylic acid or placebo respectively for 22 months on an average. There were 24 cases of reinfarction in the group of treatment and 51 in the placebo group. The difference is significant.


[Prevention of cardiovascular complications in manifest arteriosclerosis by the use of Micristine].

A long-term clinical trial in micristin-treated patients suffering from organic arterial circulatory disturbances is reported. Problems of therapy monitoring by determination of the ASA level in plasma and of control of platelet aggregation are discussed. Acute cardiovascular complications (myocardial infarction, stroke, acute vascular occlusion, amputation and angiographically demonstrated progression) were observed. The observation time did not suffice to establish statistically significant differences between micristin therapy anticoagulant treatment and basic cardiovascular therapy. The results are suggestive of a more beneficial effect of anticoagulant treatment.


[Study of the platelet aggregation inhibitor MICRISTIN as to its efficacy in the prevention of thromboembolism in the postoperative phase following surgical interventions].

Clinical test of the acetylsalicylic acid preparation micristin concerning effect and side-effect on the postoperative rate of thromboembolism in general surgery and traumatology. Prospective, randomized, checked double-blind study in 802 operated patients: 401 patients with micristin and 401 patients with placebo. Objectivization of findings in highly endangered patients by the radiofibrinogen test, ultrasonic doubler, venography of contrast medium, section. In total there were 149 thromboembolic complications = 18.6%. The placebo control group (401 patients) had 52 ensured deep venothrombosises and 8 fatal pulmonary embolisms. The group with micristin treatment (401 patients) had 23 ensured deep venothrombosises and 4 fatal pulmonary embolisms. Significant decrease of the thromboembolism rate by micristin (p = 0.001). Failures concerning the effect of micristin included fractures near the hip. Favourable effects concerned intra-abdominal surgery. Frequent side-effect: increased intra-operative and postoperative bleeding tendency.


[Effect of ASS on platelet function in experimental DIC].

In thrombin-induced DIC, acetylsalicylic acid (ASA) prevents the strong initial fall in platelet count and the obturation of the microvasculature of the lung with platelet aggregates. During the DIC reaction increasing inhibition of aggregability of circulating platelets against collagen and ADP is observed. Furthermore, ASA prevents the increase in the plasma haemoglobin level caused by DIC.


Pyrazolidine derivatives: a comparative study of their effects on platelet aggregation.

Quantitative studies were carried out of the in vitro and ex vivo effects of phenylbutazone and 3-oxoalkyl substituted diphenyldioxopyrazolidines (kebuzone, tribuzone, benzopyrazone) on platelet aggregation. The specified pyrazolidine derivatives exhibited in vitro inhibitory effects on secondary platelet aggregation (induced by adrenaline and collagen), commensurable with the effects of sulfinpyrazone. The ex vivo efficacy was markedly influenced by the height of the drug level in blood and by differences in the elimination kinetics of the pyrazolidine derivatives in human organism. Inhibitory activities against primary aggregation (induced by ADP and thrombin) were found in vitro mainly in the phenyloxoalkyl derivative of diphenyldioxopyrazolidine (benzopyrazone) and its analogues. By substitution on the phenyl attached to its alkyl side chain (for example, by a halogen in the meta position), compounds were obtained which also possessed higher activities inhibiting secondary platelet aggregation.


[Effect of aggregation inducers and inhibitors on the functional properties and the ultrastructure of platelets].

The influence of inductors and inhibitors of platelet aggregation on certain functional properties and structural peculiarities of platelets are investigated. Inductors of aggregation, such as ADP and thrombin, will cause the calcium binding to the platelet membrane to be diminished, whereas inhibitory substances of platelet aggregation will cause an increase of calcium binding. Inductors increase the number of partially degranulated platelets. Inhibitors, such as triphtazin and papaverin, inhibit these morphological changes.


[Effect of bencyclan and theophylline on changes of platelet aggregation and factor 3 activity].

The ADP- and adrenaline-induced platelet aggregation and platelet factor 3 availability were studied in patients before and eight hours after intravenous administration of bencyclan (100 mg) and/or theophylline (240 mg). Aggregation was primarily inhibited by bencyclan, the availability of factor 3 was inhibited by theophylline. Combination of both drugs exerted additive effects on both parameters. The combined use of drugs as inhibitors of aggregation is recommended, since they inhibit primary haemostasis simultaneously at two points of attack.


[Interactions of platelet inhibitors].

Combination effects of inhibitors of platelet function with different mechanisms of action such as adenosine, acetylsalicylic acid, papaverine, dipyridamole, and sodium nitroprusside were studied in vitro by means of ADP- and collagen-induced aggregation of human blood platelets. In case of ADP-induced aggregation, potentiation of the inhibitory effect was observed only with the combination adenosine-papaverine, whereas in case of collagen-induced aggregation the inhibitory effect was potentiated at various inhibitor combinations.


[Mechanism of the thrombin-platelet reaction].

In the course of investigations concerning the mechanism of the thrombin platelet interaction the conditions for the appearance of a refractory behaviour of platelets towards thrombin were examined. As investigation of the platelet reactivity by determining the amine liberation showed that a continuous or discontinuous addition of slight amounts of thrombin will cause the refractory condition. The findings were analyzed in view of the interaction between the dissolved enzyme and the fixed substrate. It is assumed that the number of receptors diminished after the first thrombin contact will cause a decrease of the receptor occupation speed on the platelet surface, thus leading to a decreased platelet reactivity.


[Preparation of human platelets without the use of anticoagulants and study of the effect of Catt on aggregation].

A method for the preparation of human blood platelets is presented which substitutes the use of anticoagulants by a gelfiltration for the removal of plasma calcium from native blood. In a second step the platelets are separated from the gel filtered blood by a centrifugation on a Ficoll density gradient. The anticoagulant-free platelets reveal an intact morphological feature and a normal aggregation behaviour in response to different aggregation inducers. It was found that in ADP-induced aggregation monophasic aggregation is shifted to a biphasic one by increasing concentrations of extracellular calcium.


[Detection of soluble fibrin monomer complexes and of platelet factor 4 in the plasma of patients with diabetes mellitus].

In 70 patients with diabetes mellitus the concentration of soluble fibrin monomer complexes and the plasma level of platelet factor 4 in blood were investigated. A significant increase of both parameter in comparison with the normal values were seen. The results indicate intravascular micro thrombosis in diabetes mellitus; the thrombocytic factors as well as the plasmatic factors of the haemostasis are included.


[In vitro studies and model calculations on the kinetics of fibrinolysis with streptokinase].

A method of measuring the distribution kinetics of radioactive 51Cr-EDTA between human plasma and a fibrinoclot are represented. In the presence of streptokinase the speed of distribution is a measure for the progressing of fibrinolysis. The results of measurement as well as the model calculations on the kinetics of fibrinolysis by streptokinase with physiologically relevant data are compared with those observations on the mechanism of thrombolysis obtained from biochemical findings. These results indicate that at the dissolution speeds of the clots observed clinically or experimentally the diffusion of streptokinase activator complexes into the clot seems to play no important role.


[The heterogeneity of streptokinases].

Purified streptokinases of some strains of different types of Strep. pyrogenes (group A) as well as streptokinases of a strain of group C or group G streptococci, resp., were studied in regard of their heterogeneity. Differences of the streptokinases were found in their amino acid compositions and in isoelectric points. Serological differences were detected by a quantitative precipitation reaction with fluorescein-coupled antistreptokinase antisera from the goat as well as by neutralization reactions of the activator activities of the streptokinases by antistreptokinase antisera obtained from guinea pigs.


[Animal experimental studies on fibrinolysis with streptokinase].

In contrast to rabbit blood plasma, in guinea pig and rat blood plasma activation of fibrinolysis by streptokinase is achieved after addition of human plasminogen or human plasma only. A simple experimental procedure for testing application forms of streptokinase in rats is described. Fibrinolysis in vivo is more effective after subsequent administration of human plasma and streptokinase in rats than after administration of a mixture of human plasma and streptokinase (activator).


[Rheologic changes with streptokinase--experimental studies].

In dogs with artificial stenosis of the femoral artery application of 96 000 IU Awelysin/kg initially and subsequent infusion of 1200 IU/kg/min caused an increase of the blood flow below the stenosis. The improvement in blood flow is attributed to the decrease in fibrinogen and to the increase in fibrinogen degradation products.


[Streptokinase therapy of myocardial infarct dependent on the severity of the disease].

A critical analysis of 14 clinical studies published since 1963 on the fibrinolytic treatment of acute cardiac infarct with streptokinase or urokinase shows that only half of them corresponds to the demands of a controlled clinical trial of therapy. The degree of seriousness of the infarct groups described in the different papers shows considerable fluctuations. The findings on letality are not uniform, there is a distinctive trend towards a reduced letality under streptokinase. It was only in recent studies that the dependence of the results of treatment on the seriousness of the disease were checked by retrospective stratification (Australian study) or by prospective stratification (European co-operative study). According to the findings of the European co-operative study those patients who are selected for the purpose of examination and affected with an infarct of medium degree of seriousness being not older than 12 hours have a significantly reduced six months letality under streptokinase on the conditions of this study.


[Successes, failures and complications in thrombolytic therapy in peripheral, arterial and venous occlusions].

Thrombocytic therapy in peripheral arterial and venous vessel occlusion represents a clearly described alternative towards the surgery of vessels. A success rate of 36.5% can be found in subacute peripheral arterial thrombosis and 46.3% in subacute thrombotic occlusion of a bypass-graft. Contrary to that, a rate of 29.8% can be found in complications or side-effects respectively. In cases of peripheral deep venous thrombosis, a partial or full success can be found in 72%. However, the rate of complication amounting to 44.2% is comparatively high. The longer thrombolytic therapy with streptokinase or urokinase will last, the more frequently and more serious will be the complications, such as bleedings of different kind as well as increase of temperature to mention the most frequent ones. The application of urokinase is absolutely possible today, however, the use of urokinase seems to be only justified, if a thrombolytic therapy with streptokinase was carried out successfully and a subsequent surgical therapy was not possible. The present costs of this preparation are far too high for urokinase to be applied routinely. A thrombolytic therapy with SK as well as with UK has to be followed by an anticoagulant treatment.


[Clinical late results after long-term treatment of acute femoro-iliacal thrombophlebitis].

The development of subfacial chronic insufficiency of veins (SCIV) was investigated after medicamentous treatment of acute femoroilliacal phlebothrombosises. Streptokinase (streptase) or heparin were used as medicaments. A control group of patients was treated without medicaments for the purpose of comparison. With the exception of the patients of the control group, all others were treated with oral anticoagulants for 6-7 months. Simultaneously, part of those patients treated with heparin were given a benzopyron preparation (venolat). After 1, 2, and 5 years a check-up was made for evaluating the stage of SCIV development. The best clinical late results could be found in patients of the streptokinase group. The effect of the combined administration of dicumarol and benzupyronan was markedly better than the sole anticoagulant treatment. SCIV developed most rapidly in patients of the control group. However, a check-up made 5 years after the beginning of therapy could not reveal the development of ulcers cruris in any patient.


[Early Awelysin therapy in massive pulmonary embolism after hip alloarthroplasty].

The fibrinolytic therapy of massive frest pneumatic embolism represents an ensured progress today. Until now it was only possible to report on those cases where the patients had survived at least two hours from the beginning of the acute event. Two cases are reported here where the patients were successfully treated in spite of a standstill in the circulation. In a 60 years old female patient and 54 years old female hypertonic patient a successful reanimation could be performed in the early postoperative phase by applying a high dosage of awelysin. The progress indicated that the therapy has led to a lysis or partial lysis of embolus respectively and thus again to a perfusion of the lungs. At the same time the serious bleeding complications clearly indicate the limits of this aggressive form of intensive therapy.


Diagnostic problems in thromboembolism.

Some pathogenetic aspects and diagnostic problems in venous thrombosis are briefly discussed. Emphasis is put on the value of blood tests. It is concluded that the various tests are neither able to detect ongoing thrombosis, nor to identify a prethrombotic state. The necessity of direct demonstration of the thrombus preferably by phlebography is stressed. The tests are valuable as a part of the "coagulation profile", and may aid to discover e.g. antithrombin III deficiency or subnormal fibrinolytic activity.


Prophylaxis of recurrent venous thrombosis with long-term enhancement of fibrinolysis.

The effects of 6 months' combined therapy with phenformin and an anabolic steroid were compared in patients with thrombophlebitis migrans (12 patients) and those with superficial thrombophlebitis (15 patients). In both groups of patients an increase in blood fibrinolytic activity, and "capacity" decrease in platelet adhesiveness, plasma fibrinogen, blood lipids, beta lipoproteins as well as serum cholesterol level were found. A statistically significant decrease in frequency of inflammations in patients with thrombophlebitis migrans occurred. In these patients a return of the previously low "fibrinolytic capacity" to normal values was observed. It seems that prolonged activation of fibrinolysis by means of phenformin and an anabolic steriod may be of value in the prophylaxis of venous thrombosis especially thrombophlebitis migrans.


[Tumor plasminogen activators and their clinical significance].

By means of a radio-immunological method the amount of plasminogen activators released by tumours was determined. The concentrations of tumour plasminogen activators was considerably higher in the venous blood of ovarial tumours than in the peripheral blood. In the uterine fluid the uterus cavity the concentration of plasminogen activator was increased in cases with neoplastic endometrium compared to those with normal endometrium.


Ia antigens in serum during different murine infections.

There exists in the mouse a family of I-region-controlled (Ia) antigens which carry carbohydrate-defined determinants. These antigens appear in serum as glycolipids and seem to be actively secreted by antigen-activated T-cells. This paper describes the ability of selected viral, bacterial, and protozoal infections of mice to markedly alter the serum levels of these Ia antigens. All the infectious agents examined induced substantial augmentation or suppression of serum Ia concentrations or both. Lymphocytic choriomeningitis (LCM) virus first enhanced and then suppressed serum Ia levels during the course of acute infection. Enhancement occurred during the time of ongoing virus replication and splenic lymphoproliferation while suppression coincided with the peak of the cytotoxic T-cell response and virus clearance. Listeria monocytogenes infection induced a substantial reduction in Ia levels at a time just after marked depletion of T-cells in the spleen. In contrast, Brucella abortus caused a significant increase in Ia levels 7 days postinfection, which correlates with the appearance of peak numbers of bacteria in tissues. Finally, Plasmodium yoelii, a nonlethal malarial parasite which stimulates prolonged T-cell proliferation, augmented serum Ia levels, whereas P. berghei, a lethal parasite which tends to inhibit. T-cell division, suppressed Ia secretion. Possible interpretations of these different results are presented.


Ability of enriched immune T cells to confer resistance in hamsters to infection with Treponema pertenue.

This investigation presents the first direct evidence that T cells are involved in resistance to challenge with Treponema pertenue. Enriched T cells from immune hamsters were obtained by sequential filtration through glass and nylon-wool columns. This procedure removed the majority of functional antibody-producing and immunoglobulin-bearing cells. The fractionated cell suspensions were less responsive to stimulation by phytohemagglutinin, lipopolysaccharide, and dextran sulfate, but they were enriched with antithymocyte-sensitive cells and were more responsive to stimulation with concanavalin A. Hamsters receiving fractionated or unfractionated immune cells had no cutaneous lesions 21 days after infection and had significantly lower lymph node weights and fewer treponemes per node than hamsters that received fractionated or unfractionated normal cells. Resistance was transferred with immune cell suspension enriched in T cells despite an absence of anamnestic antibody response to specific treponemal antigens.


Regulation and function of sucrose 6-phosphate hydrolase in Streptococcus mutans.

Sucrose catabolism by Streptococcus mutans is initiated by a phosphoenolpyruvate-dependent sucrose phosphotransferase reaction that produces sucrose 6-phosphate the latter is then cleaved by a sucrose 6-phosphate hydrolase reaction that yields glucose 6-phosphate and fructose. We have examined the regulation of the sucrose 6-phosphate hydrolase and found that it was synthesized constitutively whereas sucrose phosphotransferase activity was inducible. However, the levels of both sucrose phosphotransferase and sucrose 6-phosphate hydrolase were repressed when fructose was used as a growth substrate. The specific activity of sucrose 6-phosphate hydrolase in permeabilized cells was approximately 30 mmol/min per mg (dry weight of cells), and it had an apparent Km for sucrose 6-phosphate of 0.3 mM. analysis of a mutant that was missing sucrose 6-phosphate hydrolase activity revealed that its ability to hydrolyze sucrose was reduced.


Role of interferon in mice in protection against influenza A virus by bacterial ribosomes together with membranal glycoproteins of Klebsiella pneumoniae as adjuvant.

Nonspecific protection against infectious aerosols of influenza A virus was obtained in Swiss mice after vaccination by aerosols of bacterial ribosomes together with membranal glycoproteins extracted from Klebsiella pneumoniae as the adjuvant. It was shown that repeated stimulant aerosols were necessary to obtain this protection. Routine estimation of serum interferon levels after administration of the association of ribosomes plus membranal glycoproteins to the animals by aerosol or intravenous route showed that there was no correlation between protection and the presence of serum interferon. It was shown that the serum interferon-inducer activity was due to ribosomes. No induction of serum interferon was obtained with membranal glycoproteins used separately. Local liberation of interferon in the mucous membrane of the upper respiratory tract was not investigated.


Affinity partition of proteins in aqueous two-phase systems containing polyoxyethylene glycol-bound ligand and charged dextrans.

The partition of the delta 5 leads to 4 3-oxosteroid isomerase of Pseudomonas testosteroni in aqueous two-phase systems containing both the macroligand, polyoxyethylene glycol-bound estradiol, and charged (cationic or anionic) dextrans has been studied. If the enzyme is well retained in the upper phase by an adequate amount of macroligand, it is possible to improve the removal of the contaminating proteins by extracting them into a lower phase containing positively or negatively charged dextran. Some mult-step extraction experiments were carried out to purify isomerase, by washing the upper phase successively with cationic and anionic dextran-rich phases.


Separation of small DNA and RNA oligonucleotides by high-performance anion-exchange liquid chromatography.

Small oligonucleotides from DNA and RNA have been separated according to their base composition by high-performance anion-exchange liquid chromatography on Partisil-10 SAX using triethylammonium acetate buffer as the eluent. Fifteen of the 16 possible deoxydinucleoside monophosphates and all 16 dinucleoside monophosphates have been separated. All pairs of sequence isomers were all resolved. The 15 commercially available deoxydinucleotides were resolved into 13 fractions. A good resolution of deoxytrinucleoside diphosphates isolated from an alkaline phosphatase-Mg2+-activated DNase I digest of calf thymus DNA was achieved by this technique. A large number of sequence isomers could be fully separated. The base sequence of the eluted individual constituents has been determined by their hydrolysis with snake venom and spleen phosphodiesterase followed by high-performance liquid chromatographic analysis of the nucleotides released. The eight trinucleoside diphosphates isolated from an alkaline phosphatase-pancreatic RNase digest of yeast RNA have also been separated according to base composition. Their sequence was determined as above. The described technique is fast and gave very good separation. Most of the sequence isomers could be separated. Moreover, the eluent triethylammonium acetate can easily be removed from column effluents by freeze-drying in order to facilitate subsequent sequence analysis of the eluted compounds. The observed elution orders of the sequence isomers obey certain rules which are discussed in detail.


Study of the interaction between phosphorylase and hydrophobic groups by means of affinity electrophoresis.

A homologous series of water-soluble alkyl-dextrans varying in the length of their hydrocarbon side-chain [-NH-(CH2)n-CH3; n = 1-5] were synthesized. When alkyl-dextrans were entrapped in polyacrylamide gel, the electrophoretic mobility of phosphorylase was retarded by hydrophobic interaction between phosphorylase and the immobilized alkyl groups. The dissociation constants of rabbit brain phosphorylase, rabbit skeletal muscle phosphorylase a and b and potato glycogen and starch phosphorylases were calculated from the extent of the retardation of mobility as a function of the concentration of the alkyl groups. As the length of the hydrocarbon side-chains of alkyl groups increased, the affinity of the phosphorylases for the alkyl groups increased. The introduction of a hydroxyl or an amino group at the terminal position of the hydrocarbon side-chain diminished the affinity.


Major and modified nucleosides in tRNA hydrolysates by high-performance liquid chromatography.

We describe a high-performance liquid chromatographic analytical method that can be readily placed in operation, and which is particularly well suited to scientists investigating tRNA structure, biosynthesis, and function, and for the determination of major and modified nucleosides of tRNA. The method is characterized by the following features: (1) Sensitivity at the nanogram level; (2) High chromatographic resolution and selectivity; (3) Direct measurement of nucleosides with accuracy and precision; (4) Analysis is non-destructive and the high capacity of this chromatographic system allows easy isolation of pure nucleosides for further characterization; (5) Rapid separation and measurement in ca. 1 h after hydrolysis to nucleosides; and (6) Quantitation without use of radiolabeled compounds; however, labeled compounds are readily isolated and measured.


Rapid isolation of carcinogen-bound DNA and RNA by hydroxyapatite chromatography.

Carcinogen-bound DNA and RNA are conveniently isolated by solvent extraction and hydroxyapatite (HAP) chromatography. Tissue is suspended in 8 M urea-0.24 M sodium phosphate-1% sodium dodecyl sulfate-10 mM EDTA, pH 6.8 (MUP-SDS-EDTA) and extracted with chloroform-isoamyl alcohol-phenol (24:1:25; CIP) to remove protein. RNA and DNA are separated by passing the aqueous solution through an HAP column; RNA is eluted with MUP, DNA with 0.48 M sodium phosphate, pH 6.8. Examples presented are: (1) calf thymus DNA that has been reacted with N-acetoxy-2-acetylaminofluorene (N-OAc-AAF), (2) isolated intact rat hepatocytes incubated with N-hydroxy-AAF and (3) livers from Sprague-Dawley rats treated with N-hydroxy-AAF.


Electron microscope histochemistry of acetylcholinesterase distribution in the optic tectum of teleosts.

An ultrastructural analysis was made on acetylcholinesterase (AChE) localization in the optic tectum of two teleosts, the goldfish and the catfish. Electron microscope histochemistry reveals several details on synthesis, distribution and possible sites of utilization of the enzyme in the different tectal layers. The results show that AChE is synthesized by all the neuronal types present in the optic tectum. The final localization of the enzyme is the result of its synthesis in cell bodies, its storage and transport along dendrites and its release in extracellular spaces. The differences in AChE localization between the two teleosts examined mainly derive from differential enzyme release in the corresponding layers of the optic tectum. Cholinergic synapses cannot be precisely identified by means of AChE histochemistry, but the layers in which maximum release of enzyme in the extracellular spaces occurs most likely correspond to areas where cholinergic mechanisms are operating. In this connection some interesting differences of AChE localization in the superficial tectal layers (stratum marginale, stratum opticum and stratum fibrosum et griseum superficiale) are discussed. Electron microscopic histochemistry of AChE confirms its usefulness in better understanding some links between the anatomical and functional organization of complex neural structures.


[Kinetic determination of alpha-amylase in serum and urine with an oligosaccharide as substrate--modification for a fully mechanized enzyme measuring device (author's transl)].

The author reports a modification of the UV method UltraZyme Plus alpha-Amyl Harleco and the adaptation to the Eppendorf Enzymautomat 5010. alpha-amylase acts on an oligosaccharide mixture yielding maltose, which is hydrolysed by alpha-glucosidase. The liberated glucose is determined specifically by the hexokinase/glucose-6-phosphate dehydrogenase (NAD+-dependent) method+ by addition of pyruvate, lactate dehydrogenase and ATP. Thereafter the lactate dehydrogenase reaction is stopped by addition of oxamate and the alpha-amylase activity is measured.


[Continuously measuring procedures for the determination of alpha-amylase in serum. Comparison with amyloklastic and chromolytic methods (author's transl)].

Alpha-amylase has been measured in 57 patients' sera with normal to highly elevated amylase activities. The results obtained with four enzymatic-kinetic test-kits have been compared with those from five conventional marketed procedures. The results are evaluated by regression analysis. The specificity of the methods was analysed with amylase samples from human pancreas, human saliva and pig pancreas. The susceptibility of the kinetic procedures to interferences is smaller. The problems of conversion of activity units and reference ranges are discussed. The UltraZyme and the Monamyl neu-methods have been found superior to the other procedures because of their good practicability, precision and broad linear range; their specificity for diagnostic purposes in pancreatic diseases is less favourable compared with that of conventional procedures.


Cholesterol in high-density lipoproteins: a comparison between dextran sulfate-magnesium chloride precipitation and preparative ultracentrifugation.

The cholesterol in high density lipoproteins (HDL) has been determined in 140 serum samples after dextran sulfate-magnesium chloride precipitation of the apolipoprotein B-containing lipoproteins. The values correlated well with simultaneously measured values from preparative ultracentrifugation (slope 0.98, r = 0.93). Furthermore, there was a significant correlation in 60 specimen between the dextran sulfate-MgCl2 and heparin-MnCl2-precipitation method.


[Circadian variations of the plasma proteins in the adult male rat under natural synchronisation (author's transl)].

Circadian rhythms of blood total proteins, albumin, alpha 1-, alpha 2-, beta- and gamma-globulins were documented in 80 Wistar SPF adult male rats, synchronized by natural light (06.00-18.00 h) and darkness (18.00-06.00 h) during the month of October 78. Blood was sampled at four fixed time points in the 24 h scale (i.e. 04.00, 10.00, 16.00 or 24.00 h). Total proteins, albumin, alpha 2- and gamma-globulins showed a statistically significant rhythm with a maximum at 04.00 h. The data obtained in anaesthetized rats under artificial synchronization and in man are compared, taking into account that the rat is a nocturnally active rodent. The present work partially confirms the hypotheses of previous chronopharmacological studies, thus e.g. curarizing substances have a mimimum activity when the protein plasma level, especially albumin, is the highest.


Tumor-associated antigenic differences between the primary and the descendant metastatic tumor cell populations.

The existence of antigenic differences between cell populations in the local growth of the 3LL tumor (L-3LL) and its lung metastases (M-3LL) was studied. Normal C57BL/6 spleen cells sensitized in vitro for 5 days against L-3LL monolayers lysed preferentially L-3LL targets but not M-3LL tumor cell targets. Conversely, anti-M-366-sensitized lymphocytes killed M-3LL targets more efficiently than they killed L-3LL targets. Furthermore, spleen cells from mice bearing subcutaneous L-3LL tumors were significantly more cytotoxic to L-3LL targets than to M-3LL targets and vice versa. M-3LL cells were found also to be more resistant in vitro and in vivo to natural killer cells than were L-3LL tumor cells. M-3LL cells were more resistant than L-3LL cells to hybrid resistant mechanisms when they were inoculated into F1 (C3Heb x C57BL/6) or F1 (BALB/c x C57BL/6) mice. Anti-M-3LL lymphocytes generated both in vitro and in vivo, but not anti-L-3LL lymphocytes, admixed with L-3LL or M-3LL tumor cells and inoculated into footpads of syngeneic recipients suppressed the development of lung metastases. These results suggest that metastatic cells are indeed phenotypic variants of the local growing tumor cell populations. Presumably, these variants are selected for their capacity to home to and grow in the lungs, and for their resistance to specific immune effects initially evoked against the local tumor and to nonspecific natural killer cells. These data may prove to be of importance with respect to any rational approach to the problem of immunotherapy.


[Volvulus of the afferent loop after Billroth II operation (author's transl)].

A case report is given of a patient, who had a volvulus of the afferent loop after Billroth II operation; on relaparotomy it was found, that this loop had slipped cranially through the mesocolic slit, twisted, and become gangreneous, due to bad adjustement of the slit. Increased levels of serum amylase initially had given rise to the suspicion, that the deteriorating condition of the patient was due to pancreatitis. The patient recovered fully after resection had been done and an Y-Roux-anastomosis established. A review is given covering 22 cases from the literature.


[Whipple's disease in an elderly person (author's transl)].

A case report is given of a 73 years old woman suffering from Whipple's disease. Intermittant polyarthralgias, diarrhea, weight loss and laboratory signs of malabsorption had been present for 6 years, when diagnosis was established by biopsy: pathognomonic PAS-positive macrophages were demonstrated in the duodenal mucosa. A complete remission of the disease could be achieved after 100 mg of doxycycline had been administered continuously for a year. Some special details of the course of Whipple's disease in the higher age group are discussed.


[Antigenic markers of variable region of mouse myeloma MOPC 21 light chain].

Monospecific antibodies against V-region of the L-chain of the mouse myeloma protein MOPC21 (VK 15 group) were obtained and used for preparation of antibody-immunoadsorbents. By means of the immunoadsorbents the content of normal immunoglobulins with antigenic markers of VK 15 group in the serum of mouse strains BALB/c, CBA and C57BL/6 was determined. The least quantity of the proteins determined by this method was estimated to be 1.4%. If each of K-chains of the variable groups is represented equally in the population of normal immunoglobulins, the number of VK groups and respectively germ-line VK genes should be not less than 70.


[Inhibition of RNA synthesis in vitro by antibodies against mononucleotides].

Antibodies against purine nucleotides were obtained from rabbits immunized with conjugates of bovine serum albumine with AMP or GMP. The antibodies purified by affinity chromatography on nucleoside monophosphate-human serum albumine-Sepharose columns inhibited RNA synthesis on native T4 phage DNA by E. coli RNA polymerase. The inhibition of transcription was due mainly to inhibition of the initiation stage of RNA synthesis.


Growth, morphology, and function of xenotransplanted human tumors.

Tansplantability, growth, morphology, and function of xenotransplanted human tumors, such as carcinomas of the lung, liver, breast, choriocarcinoma, and blastomas of the liver, lung, kidney, and uterus, are described. From the tumor take rate, it is clear that xenotransplantation cannot be used for the study of every human tumor: slow-growing tumors are difficult to analyze, and functioning adenomas and low-grade malignant carcinomas are at present almost impossible to study by this approach. From the authors' transplantation experience, tumor antigenicity to nude mice with no T-cell function, either tumor specific or species specific, was suspected. Therefore, the growth in nude mice may not equate to that in the human body. The stroma of the transplanted tumor, which is most likely of mouse origin, might also alter the growth rate, as it did the histology of some tumors. Another possible hindrance that has not been described in the text is the mouse endogenous virus. Serially transplanted human tumors are often infected with C particles, which could well influence the tumor growth and character. In spite of the presence of some factors unfavorable for the study of human tumors through xenotransplantation, it has, nevertheless, been clearly shown that the nude mouse/human tumor system is a very useful tool for functional analysis of tumors in relation to growth, differentiation, and morphology, such as eutopic or ectopic production of various hormones, AFP, normal serum proteins, colony-stimulating factor, erythropoietin, and so on. This system can be employed to elucidate the production of many other biologically active and inactive substances by a variety of tumors and their effects on the host in the future and should provide better understanding of human cancers. Attempts to induce differentiation and to change the biologic behavior of xenotransplanted human malignant tumors have failed so far, except for induced dormancy of breast carcinoma under unfavorable hormonal conditions. This line of investigation may have particular import on cancer research, particularly in relation to the biology and treatment of human cancers.


Recognition of structure of 5-halogenated derivatives of ara-UTP by DNA polymerase gamma and reverse transcriptase.

This report deals with the test of a series of 5-halogenated derivatives of ara-UTP for the inhibition of DNA polymerase gamma and viral reverse transcriptase. The compounds newly synthesized and tested were; ara5-FUTP, ara5-C1UTP, ara5-BrUTP and ara5-IUTP. The results were: 1) All these compounds were inhibitory to DNA polymerase gamma and reverse transcriptase. The mode of inhibitions was, in all cases, competitive against dTTP. 2) Ki values for these inhibitors with DNA polymerase gamma were lower than those with reverse transcriptase. 3) Ara5-ClUTP was less inhibitory to reverse transcriptase than other derivatives.


Prostaglandin synthesis inhibition and monoamine metabolites in the rat brain.

The effect of indomethacin 3 mg/kg on levels of homovanillic acid (HVA), 4-hydroxy-3-methoxy phenyl ethylene glycol (HMPG) and 5-hydroxy indol acetic acid (5HIAA) was studied in rat striatum and olfactory tubercle with and without pretreatment with morphine 10 mg/kg. Indomethacin caused a small decrease in resting levels of HVA in striatum but not in olfactory tubercle. No effects were seen on resting or morphine induced changes in the levels of these monoamine metabolites. Likewise indomethacin 20 mg/kg failed to alter the elevation of HVA induced by chlorpromazine 15 mg/kg or the decrease of HVA induced by apomorphine (1--10 mg/kg) in the rat striatum. Our results do not support a major role for endogenous prostaglandins in the modulation of monoamine neurotransmission in the rat brain.