Modifications by stretches of the mechanical response of isolated cerebral and extracerebral arteries to vasoactive agents. Length-tension relationship was compared in helically cut strips of canine cerebral, coronary, mesenteric, renal, and femoral arteries. Tension developed progressively by increasing the stretch; with the same strain, a greater passive tension developed in cerebral than in extracerebral arteries. The peak active tension developed by serotonin (cerebral, coronary), norepinephrine (mesenteric, renal and femoral) or K+ (coronary) was obtained at a resting tension of 1.5 g (other than coronary) or 2.0 g (coronary). Papaverine (10(-4) M) caused a relaxation in cerebral arterial strips contracted with serotonin to a level lower than that prior to the addition of serotonin, the relaxation from the initial level of tension being related directly to the stretch applied. The relaxing effect of adenosine was related directly to stretches of cerebral arterial strips. It seems likely that a rise of intra-arterial pressure effects a greater increase in the wall stiffness in cerebral than in extracerebral arteries. The responsiveness to vasoconstricting and vasodilating agents of both cerebral and extracerebral arteries appears to increase when the arteries are distended.


Current-voltage relations in ventricular muscle preparations from different species. Current-voltage relations were determined in ventricular muscle preparations from dog, sheep, pig, guinea pig, bull and cat hearts. The single sucrose gap voltage clamp method was used to apply 2 s steps or slow speed voltage ramps (3-4 mV/s). 1. The current-voltage relations obtained with 2 s steps were similar to those obtained with ramps. 2. Negative slope regions were readily apparent in the current-voltage relations of preparations from all species except cat although the latter did display marked anomalous rectification. 3. Increasing the external potassium concentration from 3 to 10 mM increased the slope of the current-voltage relation around the resting potential and resulted in the "crossing-over" and "crossing-back" of the relations if all tissues. 4. The two extreme cases of bull (marked negative slope region) and cat (no negative slope region) were compared with regard to transgap intracellular and extracellular resistances; they were quite similar. This suggests that the current-voltage relations reflect differences in membrane properties rather than, for example, different proportions of non-membrane leakage current. 5. In bull ventricular preparations, the application of D-600 (2 X 10(-6) M), 30 min) had only a small effect on the current-voltage relation. In contrast, cesium (10-20 mM) nearly abolished the negative slope region suggesting that this characteristic of the current-voltage relation is due to potassium channel rectification. 6. Possible explanations for action potential durations in bull ventricular fibres being 50 to 100% longer than in cat are considered.


Respiratory oscillations of the arterial PO2 and their effects on the ventilatory controlling system in the cat. The respiratory oscillations of the arterial PO2 were measured in paralyzed, artificially ventilated cats by a small (1.2 mm) fast-responding catheter oxygen electrode. The amplitude of these oscillations could be changed independently of the mean PA,O2n by a specially designed respirator circuit. deltaPaO2 was shown to increase with increasing tidal volume or decreasing frequency of the respirator, and with increasing mean PaO2. The amplitude of the oscillations was attenuated considerably from the left atrium to the aorta. No attenuation occurred from the aorta to the carotid artery, provided that the blood flow in the carotid artery was not impeded. The measured attenuation of the oscillations was compared to that calculated by Yokota and Kreuzer (1973) and found to be quite different. The output of the ventilatory controlling system of the cat was measured from the quantified phrenic nerve activity. When only deltaPaO2 was changed at a constant level mean PaO2, the quantified phrenic nerve activity did not change, indicating that the amplitude of the oscillations does not influence the ventilatory controlling system. In vagotomized animals, the periodicities of the oscillations and the phrenic nerve activity were completely dissociated. From the fact that no Cheyne-Stokes type of breathing occurred, it was concluded that the effect of timing is negligible.


Thermally-induced cutaneous sympathetic activity related to blood flow through capillaries and arteriovenous anastomoses. In the ears of anaesthetized rabbits cutaneous efferent sympathetic nerve activity (SkNA) and blood flow (Q) to capillaries have been measured during various thermal treatments. Warming the spinal cord or skin of the body midside caused a marked decrease in SkNA but capillary Q increased only slightly. Exposure to a warm environment or localized warming of the ear alone induced either a decrease, an increase, or no change in SkNA, but capillary Q always increased markedly. The usual slight increase in capillary Q during spinal warming, was abolished by preventing the usual marked increase in skin temperature. When the spinal cord of the conscious rat was warmed, a marked increase in temperature of the tail (which contains arteriovenous anastomoses, AVA's) indicated dilatation, whereas there was no change in ear temperature (where there are no AVA's). When these results are considered together with recently defined differential influences of reflex and direct effects of temperature on blood flow through cutaneous AVA's and capillaries, it is concluded: (1) That thermally-induced reflex changes in skin blood flow are mediated via sympathetic nervous action on AVA's; (2) Changes in blood flow evoked by direct heating take place through the capillaries, not the AVA's, quite independently of SkNA.


Intracerebral gas partial pressure changes under vasoactive drugs. A mass spectrometry study. An original technique based on mass spectrometry has been used to measure simultaneously in undrugged, freely-breathing animals local PO2 and PCO2 in the brain (caudate nucleus) and arterial PO2 and PCO2 in the aorta. Injection of vasoactive drugs brought about significant modifications of the partial pressures of the physiological gases in the brain. The origin of these variations was discussed, based on the combination of 3 possible factors: cerebral vascular changes, systemic metabolic changes (PaO2 and PaCO2) and local metabolic changes. It was demonstrated that local variations of PCO2 cannot be held to be responsible for the modifications of cerebral blood flow induced by papaverine or the sympathomimetic drugs.


Effects of locally applied chemicals on transretinal potential and horizontal cells in the isolated carp retina I. Amino acids. Experiments were performed on isolated retinas of teh carp (Cyprinus carpio) to investigate the effects of locally applied amino acids on the transretinal potential and the membrane potential of horizontal cells. A minute quantity of each solution of an amino acid was applied locally at different layers by means of pressure-microinjection. The responses were induced by alternating central and annular stimuli of white light. Amino acids applied locally at the distal layer (receptor surface or outer plexiform layer) in the centrally stimulated area, caused, at concentrations of 25--50mM, rapid changes in the light-induced potential, whereas those microinjected into the inner plexiform layer did not. When applied locally at the distal layer, amino acids abolished the proximal PIII response and reduced the amplitude of S-potentials, particularly in response to a central stimulus. However, the resting potential of horizontal cells was depolarized only slightly by acidic amino acids. The results suggest that the passive membrane properties of horizontal cells are not altered significantly by the amino acids applied. Differential susceptibilities of the PII component to acidic and neutral amino acids were found; the former did not change or, in some cases, enlarged, whereas the latter abolished it. Possibly, neutral amino acids interfere with a neural mechanism generating the PII component.


Asymmetry of the chloride transport system in human erythrocyte ghosts. The concentration dependence of the unidirectional chloride flux and the inhibition of the unidirectional chloride flux by sulfate were studied in human red cell ghosts. The concentration dependence of the unidirectional chloride flux and its inhibition by sulfate were asymmetric. The unidirectional chloride flux can be saturated from the inner and from the outer membrane surface. For the inner membrane surface, lower chloride half-saturation constants were obtained than for the outer membrane surface. The inhibition of the unidirectional chloride flux by sulfate is competitive. In contrast to the chloride half-saturation constants, the inhibition constants of sulfate for the inner membrane surface were higher than the inhibition constants of sulfate for the outher membrane surface. Either there are fixed anion binding sites at the inner and at the outer membrane surface which control the access of anions to a pore, or there is a mobile carrier which is in contact with both membrane surfaces. The asymmetry of the concentration response and of the inhibition of the unidirectional chloride flux suggest that the anion binding sites at the inner and at the outer membrane surface differ with respect to their affinities for chloride and for sulfate. Alternatively, the asymmetry of the chloride transport system could indicate an asymmetric distribution of a mobile anion carrier across the erythrocyte membrane.


[Treatment of insulin-resistant form of diabetes mellitus by intravenous droplet infusion of insulin]. Four patients with an insulin-resistant form of diabetes mellitus were treated with intravenous droplet injections of swine insulin. There was revealed a rapid reduction of the 24-hour requirement in insulin and of the antiinsulin antibody titre. The method proved to be effective for the treatment of patients with "immune" and "nonimmune" forms of insulin resistance. The method is recommended for the treatment of patients with diabetes mellitus decompensation.


Dopaminergic mediation of beta-endorphin-induced catalepsy. Acute intracisternal administration of human beta-endorphin produced catalepsy and increased striatal concentrations of 3,4-dihydroxyphenylacetic acid (DOPA) and homovanillic acid (HVA). All of these effects were blocked by naloxone. Apomorphine, a dopamine receptor antagonist, also prevented beta-endorphin-induced catalepsy and the increase in striatal DOPAC and HVA. The combination of subcataleptic doses of haloperidol and beta-endorphin produced catalepsy and large increases in striatal DOPAC and HVA. These data provide evidence for a role for nigrostriatal dopamine neurons in beta-endorphin-induced catalepsy. The apparent increase in striatal dopamine turnover following beta-endorphin administration may be compensatory.


Pathways of propranolol metabolism. Use of the stable isotope twin-ion GC-MS technique to examine the conversion of propranolol to propranolol-diol by 9000g rat liver supernatant. The metabolic conversion of propranolol to propranolol-diol [1-(1-naphthyloxy)-2,3-propyleneglycol] by rat liver 9000g supernatant was demonstrated to proceed through the intermediate 3-naphthyloxy-2-hydroxypropionaldehyde. Using side chain deuterated propranolol-d5 as substrate, propranolol-diol-d4 was produced, indicating an obligatory aldehyde intermediate. Analysis of desisopropylpropranolol obtained in the same incubation showed no loss of deuterium indicating that the possibility of rapidly tautomerizing imine intermediate did not make a significant contribution in this metabolic conversion. Desisopropylpropranolol, added in small amounts to the incubation mixture is more rapidly converted to the propranolol-diol than is propranolol, indicating that the major pathway of conversion of propranolol to its diol metabolite is through the desisopropylpropranolol intermediate.


Plasma vitamin A transport and visual dark adaptation in diseases of the intestine and liver. Quantitative analyses of plasma concentrations of retinol binding protein (RBP), prealbumin and total proteins were performed in normal subjects and in forty-two patients suffering from diseases of the intestine and liver. The visual dark adaptation ability (DAA) was also assessed. Reduction of DAA and of RBP and prealbumin levels was noted in patients with chronic liver disease and fat malabsorption. In sixty-eight patients with intestinal diseases the RBP concentration seemed to be reduced in relation to the degree and duration of steatorrhoea. Furthermore, inflammatory activity, as revealed by laborabory tests, markedly reduced the RBP level. Treatment with vitamin A increased the RBP and prealbumin concentrations and restored the DAA to normal in patients with malabsorption but normal liver function. In patients with liver disease reduced DAA and serum RBP values were not affected by vitamin A therapy. Only at RBP concentrations below half the normal was impairment of the dark adaptation observed, suggesting that serum RBP is a more sensitive indicator of vitamin A deficiency than measurement of dark adaptation.


Phenylalanine depletion for the management of phenylketonuria: use of enzyme reactors with immobilized enzymes. Multitubular enzyme reactors with immobilized phenylalanine ammonia lyase were tested in vitro and in vivo for depletion of phenylalanine in circulating blood. Sustained reduction of phenylalanine was achieved in less than 30 minutes. A 50% decrease of phenylalanine was obtained with a 2-hour application of enzyme reactors and was maintained for more than 2 days. Similar enzyme reactors have therapeutic potential for temporary management of phenylketonuric patients when their circulating phenylalanine becomes exceedingly high because of infection, fever, or pregnancy.


Formation of mutagens in beef and beef extract during cooking. Mutagens, distinguishable from benzo[a]pyrene and from mutagenic amino acid and protein pyrolysis products, are formed when ground beef is cooked in a home hamburger cooking appliance or when beef stock is concentrated, by boiling, to a paste known commercially as beef extract. "Well-done" hamburgers contain about 0.14 part per million of the mutagens, and beef bouillon cubes which contain beef extract about 0.1 part per million. Since such mutagens may be potentially carcionogenic and are formed during ordinary cooking procedures, their occurrence raises questions about possible risks to human health.


Histidine-rich protein as a model malaria vaccine. Ducklings were successfully immunized against Plasmodium lophurae with a purified and characterized histidine-rich protein as antigen. The use of adjuvant is not required for this protective effect, and immunity can be passively transferred with serum.


Phagocytosis in the retinal pigment epithelium of the RCS rat. The retinal pigment epithelium of RCS rats, previously thought not to phagocytize photoreceptor outer segments, exhibited a peak of phagocytosis in vivo when animals were kept under conditions of cyclic lighting (12 hours of darkness and 12 hours of light). The peak occurred at 1 hour after the onset of light, with maximum and minimum levels of phagocytosis averaging about 5 percent of that found in  the pigment epithelium of Osborn-Mendel rats used as a control. Eyecups that were obtained from Osborn-Mendel rats and maintained for up to 3 hours in organ culture demonstrated levels of phagocytosis that were sevenfold greater than those of unincubated controls. Likewise a tenfold increase occurred in incubated as opposed to unicubated RCS eyes, raising the possibility that phagocytosis could be experimentally stimulated in vivo.


Cutaneous angiosarcoma of the hip.

Cutaneous angiosarcoma of the hip. An elderly patient is presented who developed a fatal angiosarcoma of the hip about ten years after surgery and radiation for a uterine malignancy. She developed a metastasis of the uterine tumor, requiring additional radiation, which led to aseptic necrosis of the hip and chronic joint and soft tissue infection of the hip from a joint prosthesis. Subsequently she developed chronic lymphedema of the hip and finally the cutaneous angiosarcoma in this area. Any or all of these factors might bear a causative relationship to the angiosarcoma, but none can be proved.


Bilateral renal loss resulting from intravesical formalin instillation. A patient was referred for evaluation of obstructive uropathy after treatment with 10% formalin to control recurrent hemorrhagic radiation cystitis. Progressive bilateral ureteral, pelvic, and renal parenchymal destruction occurred despite ileal ureteral replacement. This complication of therapy reinforces the concept that vesical instillation of formalin should be done only in extreme cases and then only in low concentrations.


Purpura and Schamberg's disease.

Purpura and Schamberg's disease. We have described a 9-year-old boy with progressive pigmentary dermatosis. The disease requires no treatment, but must be considered in the differential diagnosis of purpura in childhood to avoid unnecessary delay and concern to the patient.


Euploid somatic recombinants with two active X or XY (1)Y(I) chromosomes isolated from cultured male Indian Muntjac cells after HVJ virus fusion, and their use for gene assignment. Four diploid somatic recombinants were isolated from hybrids either between or within two diploid cell lines of a male Indian muntjac after HVJ virus-mediated cell fusion. Both parental lines had a normal male karyotype, 7,X,Y1,Y2, in which the largest autosomal pair was heteromorphic with respect to the size of the secondary constriction (1h+/1h-), C bands, and nucleolar organizers. Of the four recombinants, three showed a 6,XX,1h+/1h+ or 1h+/1h- karyotype, the remaining one a 7,XY1Y2,1h+/1h+. No late-replicating X chromosome was found in the XX recombinants, although it was demonstrated in the natural XX line, suggesting the presence of two active X chromosomes in the former. The G6PD, PGK, and HPRT activities were proportional to the number of active X chromosomes present in all cell types examined, indicating their X-linkage, whereas the same level of activities obtained for LDH and 6PGD indicated their autosomal linkage.


Phenotypic expression of malignancy in hybrid and cybrid mouse cells. This study has been directed toward the effect of cytoplasmic transfer on the expression of marker properties in hybrid cell systems. Conventional hybrids between two nucleated cells were constructed between tumorigenic and nontumorigenic cells. Cytoplasmic hybrids, or cybrids, were constructed between enucleated chloramphenicol resistant (CAP R) donor cells (cytoplasts) and nucleated recipient cells. Clear-cut evidence for the cytoplasmic transmission of CAP resistance was obtained. Although cytoplasmic transfer had no effect on tumorigenicity or growth in soft agar, preliminary evidence was found that saturation density of the recipient cells could be altered by cytoplasmic addition in cybrids.


The relationship between induced mutation frequency and chromosome dosage in established mouse fibroblast lines. The frequencies of induced mutation to ouabain (OUA) and 6-thioguanine (6TG) resistance were compared in two established mouse fibroblast lines with different doses of the active X chromosome. The activity states of the X chromosomes were determined by DNA replication studies. Mutation frequency to 6TG R, an X-linked recessive phenotype, was inversely related to dosage whereas OUA R, a codominant phenotpye, occurred with equal frequency in both lines. 6TG R clones isolated from the line containing a majority of cells with two active X chromosomes were monosomic for the active X chromosome. Enzyme activity studies of these cytologically monosomic lines yieled results that were consistent with the presence of a single active X chromosome. The findings strongly support the hypothesis that mutation frequency is dependent on gene dosage.


[Serological typing of isolates of Pasteurella multocida from poultry and other animals in the Netherlands (author's transl)]. Pasteurella multocida isolates, isolated from poultry, birds and mammals in the Netherlands, were typed serologically. The standard sera were directed against Heddleston reference strains. Of the 202 isolates, 37.6 per cent were of type 1, 24.2 per cent of type 3, 2 per cent of type 5, 5.9 per cent of type 10 and 3.5 per cent of type 12. Cross reactions with two serotypes were shown by 18.8 per cent of the other isolates and with three serotypes by 3.5 per cent. The serotype of P. multocida isolates from several animals of the same flock on three farms was determined. Practically all the isolates from one and the same farm were found to be of the same serotype. The resulting prospects of serotyping for the use of vaccines are referred to.


[Radiological and clinical considerations on navicular disease (author's transl)]. The radiological and clinical interpretation of alterations of the navicular bone, the interaction of radiological and clinical symptoms in relation to the age of the animal, the influence of work-load on this interaction and the possibility of predicting navicular disease at an early stage are discussed. The discussion is based on experience in a study of 130 horses.


Antimycobacterial activity of some potential chemotherapeutic compounds. Fourteen compounds were tested in vitro for activity against Mycobacterium intracellulare and other pathogenic mycobacteria. Only clofazimine and chaulmoogric acid showed significant activity against M. intracellulare. In view of known minimal side effects of clofazimine further studies are warranted for this drug in chemotherapy of M. intracellulare infections.


Severe hemolytic disease of the newborn associated with anti-JSb. The red blood cells of the group A infant of a group O central African Negro woman of the Zezuru tribe with anti-Lea and anti-JSb in her serum were found to be strongly agglutinated by a commerical antiglobulin reagent six days after birth. The high concentration of bilirubin in the infant's serum gradually decreased but he became profoundly anemic and, in the absence of a suitable donor, was successfully transfused with blood from his mother. Subsequent tests using the maternal anti-JSb diluted one in ten to avoid agglutination by the anti-Lea showed that the frequency of the JS(b-) phenotype in group O Zezuru is approximately one per cent.


[Effect of cycloheximide on the ultrastructure and several metabolic processes of the cells of an SPEV culture. II]. The action of cyclohexamide (in doses of 1 and 10 mkg/ml in the course of 24 hours) on porcine embryo kidney cells in culture is accompanied by a powerful suppression of protein and DNA synthesis, mitotic activity, a weaker suppression of RNA synthesis, a lowering of the activity of succinate-, lactate- and alpha-glycerophosphate-dehydrogenase, which leads to disorders in the ultrastructure of the cells. After incubation of cells in a fresh medium (in the course of 18, 24 hours) there occurs a total restoration of the ultrastructure of the nuclei, granular endoplasmatic reticulum, mitochondria, due to the repairing and strong intensification of synthetic processes, respiration and glycolysis, mitotic activity of the cells.


[Frequency of lethal chromosomal and genome mutations in man]. According to the cytogenetic studies chromosome anomalies frequency in 209 children who died in the perinatal period was 7.2 per cent. These cases were found to consist of 8 autosome trisomies, 5 anomalies in sex chromosome system, 1 case of triploidy and 1 case of structural reorganization. Accounting for the elimination of chromosome anomalies during the early periods of fetus development, the frequency of chromosome lethals was shown to be 6.44 X 10(-2) in registered pregnancies and 54.89 X 10-2 in conceptions.


[Quantitative changes of the cytotrophoblast in underweight and normalweight newborn after risk pregnancy and antepartal therapy (author's transl)]. A histochemical-morphometric method was used to determine at the cytotrophoblast of 73 placentas, changes in the composition of the syncytium and in the Langhans' cell count in underweight and normalweight newborn with or without antepartal maternal therapy in comparison to newborn after uneventful pregnancy. The reduction of the percentage share of mature syncytium when the share of predegenerative and degenerative syncytium is increased, as weil as the increase in the average Langhans' cell count in the placentas of premature infants, hypotrophic and eutrophic infants delivered at term after risk pregnancy, compared with normal controls, is statistically significant. Significant differences in the percentage share of mature syncytium between the placentas of premature infants and hypotrophic infants delivered at term, as well as between placentas of eutrophic infants delivered at term, depending of the nature and severity of the risk, are discussed in connection with the significance of the syncytium for the processes of material transport. No correlation with the Apgar score and the occurrence of the respiratory distress syndrome of newborn, nor with the duration of the first and second stage of lavor with macroscopic placenta peculiarities, have been found. The good correlation between placenta findings and classification characteristics of newborn, and clinical aspects of the course of pregnancy, allows the possibility of diagnostic evaluation. Our results indicat also that the morphological performance must be included in the considerations explaining the success of treatment of placental insufficiency.


[Effect of amino acids on transformation of glucose oxidase as antigen in tissues of immunized animals]. The effect of certain amino acids on transformation of glucoseoxidase as an antigen in different tissues of the animals immunized with it showed that the used glucoseoxidase of the fungus Penicillium vitale Pidopl. et Bilaj possesses antigenic properties peculiar to this enzyme isolated from other sources. 1.5 minutes after a single administration of the antigen to nonimmunized rabbits it is determined in them in descending amounts in such an order: blood, lungs, liver, kidneys, lymphatic nodes. The same order is for immunized animals, but the enzyme quantity 1.5 minutes after the last injection is lower considerably in the kidneys, spleen, lymphatic nodes and in blood serum and changes slightly in the lungs and liver. The preliminary loading of animals under experiment immunized with amino acids accelerates essentially disappearance of the antigen under determination (glucoseoxidase in the given case) in the studied tissues. The data obtained testify to the fact that acceleration of antigen disappearance may be not related to interaction with antibodies.


[Experiences with ambulatory anesthesia in urology]. On the basis of more than 6000 cases is reported on the daily ambulatory practice in anaesthesia carried out on urological patients for several years. The problems and the necessity of the ambulatory anaesthesia, methods and experiences are discussed and it is referred to the errors and danger sources. Exact indication, better instruction of the patients and the parents (in children) on the consequence of the ambulatory anaesthesia, exact observation of the pre- and postoperative precautionary measures, strict organisation, choice of a suitable anaesthetic and adequate methods of anaesthesia are according to our experiences the most important prerequisites for a smooth course and an anaesthesia without complications in ambulatory surgical interventions.


[Focal nodular hyperplasia of the liver (author's transl)]. Focal nodular hyperplasia of the liver in a patient with hypernephroid carcinoma of the kidney is reported. The histological and the angiographic differentiation to adenoma and carcinoma of the liver and the therapeutic procedures are discussed.


[The value of special care in risk pregnancies in order to prevent low birth weight. 1. Results of treatment]. In this paper the results of special monitoring of pregnancies with risks are reported and compared with a collective of not specially monitored pregnancies with risks. It is possible to reduce the total rate of low birth weights and the rate of premature infants significantly by special care with consequent use of a schedule of monitoring. The rate of premature infants was 3.7 per cent in the specially monitored collective as far as the women were accessible to a prolongation of gestation. In the cases of birth weights up to 2500 g a significant removal towards higher and more favourable birth weights can be achieved by special monitoring. The causalities in the premature infants are demonstrated. The prolongation of gestation by cerclage is to be considered as a partial success.


[The value if special care in risk pregnancies for prevention of low birth weight. 2. Studies on effectiveness]. The rate of pregnancies with risks is about 50 per cent in our area. A retrospective examination of 906 pregnancies with risks and 185 analysed cases of low birthweight infants of the years 1974 to 1976 shows, that 79 per cent of the low birth-weight infants can be predicted by the use of the catalogue mentioned in the first part. The comparison of the percentage of risk factors in the two collectives indicates, that the group of pregnancies with risks is representative. In connection with the results reported in part 1 a special monitoring of all pregnancies with risks is considered to be justified. The causes for success are pointed out. If there is no possibility to organize special monitoring, the consequent use of the schedule of monitoring by trained specialists is recommended.


[The problem of premature births in the GDR]. In a 5,5% sample of the lifeborns in the GDR in 1974 the distribution of the rate of prematures by uniquebirthes is evaluated for child bearing age, birth-order and age and order at the same time. The sample points a minimum in the group of the 2nd order and age between 25 and 30. Undependently of this result the distribution of the rate of prematures by twins is evaluated for the order of birthes and borns and for child bearing age. The influence of the twins on these distributions is discussed. Modifications of the human reproduction as a result of the decontrol of abortion in 1972 are discussed in its consequences of the rate of prematures. In consequence of the decontrol of abortion an ascent of the rate of prematures would be expected without patronize the risk for premature directly.


[Relationship between employment and the birth of low-birthweight infants]. In the period from 1969 to 1976 there born 14.368 living single infants with known duration of pregnancy in the gynecological clinic of the district hospital of Schwerin.--By means of this material was proved, if there is a relation between professional activity and the birth of "infants of low birth weight".--For some professional groups such a relation could be demonstrated.


The choice of test for diagnosis of venous thrombosis. Doppler ultrasound is the preferred screening test in asymptomatic patients with high risk for venous thrombosis. Radiographic phlebography leads to definitive diagnosis in most instances. Radionuclide angiography using 99mTc MAA with delayed images to detect particle entrapment in venous blood clots can be used when radiographic phlebography is contraindicated or impractical. To detect active thrombosis in patients with past venous disease, the serial fibrinogen uptake test is the method of choice. This method is also used to monitor the efficacy of anticoagulation therapy and to detect propagating thrombosis in patients failing to respond to anticoagulation, hopefully before massive lethal pulmonary embolism occurs. Radionuclide methods are contraindicated in pregnant women and children where non invasive methods are preferred. No completely satisfactory test exists for detection of hypogastric vein thrombosis. Among the methods currently being evaluated in the laboratory the ultrasound B-scan imaging, 99mTc MAA uptake test and 99mTc mAA venous scan offer the best possibilities for successful clinical application.


[Changes in the metabolism of cerebral mitochondria under the influence of sera from schizophrenic patients and their relatives]. With the aid of a polarographic method it was demonstrated that the blood serum of schizophrenic patients and their relatives, unlike the blood serum taken from normal donors evoke a significant change of the metabolism in the mitochondria of the hemispheric cortex. The intensity of oxidation and respiration control under the influence of these sera change similarly, while the intensity of phosphorylation and oxidation with phosphorylation coupling change significantly to a higher degree under the influence of the blood of probands. Changes in the metabolism indices of mitochondria under the influence of these sera to a significant extent are genetically determined.


[Changes in neurons of a spinal ganglion culture during chronic exposure to increasing concentrations of morphine]. On the model of spinal ganglion cultures in mice by means of a light and electron microscopy the authors studied neuron changes in chronic influence of increasing concentrations of morphine. These studies demonstrated changes in most of the cytoplasmatic organells: changes in the granular and agranular reticulum; an appearance of a large amount of small vesicles in the cytoplasma and lysosome; changes of the mitochondria (partial vacuolization of the crysts and appearance of a big amount of small mitochondria); a large amount of myelin-like structures; changes of the nuclear membranes; nucleoli and an increased amount of ribosomes.


[Myelin toxicity of the serum of patients with amyotrophic lateral sclerosis in nerve tissue cultures]. In order to study the myelinotoxicity of the blood serum in patients with lateral amyotrophical sclerosis the authors used myelinated explants of the spinal cord of mice embryo. A total amount of 17 blood sera of patients with lateral amyotrophical sclerosis was studied. It was demonstrated that the serum of such patients introduced into nourishing medium in a concentration of 25% does not lead to an expressed demyelination of the nervous fibres. Cultivation of the spinal cord in a medium, containing 5--25% of the patient sera from the moment of explanation does not influence the process of myeline sheath formation. These data indicate that the serum of patients with lateral amyotrophical sclerosis do not possess myelinotoxic and myelininhibiting properties.


[Electrophoretic patterns of differently prepared fibrinogen subunits in sodium dodecyl sulfate containing polyacrylamide gels]. The electrophoretic behaviour of mercapto, carboxamidomethyl, carboxymethyl and thiosulfonic acid derivatives of rabbit fibrinogen subunits was investigated electrophoretically in sodium dodecyl sulfate containing polyacrylamide gels. Comparing carboxymethyl, carboxamidomethyl and thiosulfonic acid derivatives with the corresponding mercapto subunits divergent electrophoretic patterns were observed. Especially, the position of the Bbeta-chain was strongly dependent on the method of preparation. Similar results were obtained from investigating electrophoretic mobilities of albumin with differently substituted SH-groups after reduction with mercaptoethanol.


Serum zinc concentrations in Finns.

Serum zinc concentrations in Finns. Serum zinc concentrations were measured in 1416 Finns aged 15 years and above and belonging to 18 population groups living in various parts of the country. The mean serum zinc concentration was for men 12.7 mumol/l (82 microgram/100 ml) and for women 11.7 mumol/l (77 microgram/100 ml). Zinc levels varied with sex, age, length of fast, time of day and geographic area. It is noted that the incidence of coronary heart disease, the occurrence of a number of elements in soil and water, and the zinc level in human sera can in a statistical sence be linked to soil composition in Finland. Causal relationships, however, remain obscure.


Hypertrophic cardiomyopathy in the aged. Although usually considered a disease of young or middle-aged adults, hypertrophic cardiomyopathy is not infrequently seen in older patients as well. Twenty of 23 cases of hypertrophic cardiomyopathy seen in the past 2 1/2 years at our institution have been in patients whose average age was 65 years, and who ranged up to 76 years. Sixteen of these had evidence of an obstructive component at cardiac catheterization or echocardiography. Symptoms and signs were similar to those described for the younger patients in the literature, but were often attributed to other causes, including valvular aortic stenosis, arteriosclerotic or hypertensive heart disease, or cerebrovascular disease. Left ventricular hypertrophy was more consistently present on ECG than on x-ray. The not-infrequent occurrence of hypertrophic cardiomyopathy in older patients, predominantly females, indicates that the natural history of this disease includes a group who suffer few or no symptoms until late in life. Clinical management of younger patients with this diagnosis should be considered in light of this more favorable possible course.


Newborn head size and neurological status. Predictors of growth and development of low birth weight infants. One hundred eighteen infants with birth weights of 2,000 g or less were evaluated for growth and development at approximately 5 years of age. Review of neonatal data indicated that head circumference less than the tenth percentile at birth and abnormal neurological examination in the newborn period were important predictors of outcome. Both of these factors were associated with poor growth, later microcephaly, and neurological deficit. In addition, neonatal microcephaly was substantially related to poor intellectual attainment. Other associations were seen between microcephaly at birth and intrauterine growth retardation, low Apgar score, and low socioeconomic status, factors that also correlated with poor outcome.


Respiratory dysfunction in thrombotic thrombocytopenic purpura. Thrombotic thrombocytopenic purpura is a rare disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, neurologic abnormalities, fever and renal dysfunction. in six of seven consecutive patients with thrombotic thrombocytopenic purpura seen in an eight month period, respiratory impairment was present. Respiratory dysfunction was characterized by tachypnea, hypoxemia nad infiltrates on chest roentgenogram. Five patients required mechanical ventilation. Two patients had cardiogenic pulmonary edema, but they remained hypoxemic despite treatment for pulmonary edema and maintenance of normal pulmonary capillary wedge pressure for more than 36 hours. Four patients died and autopsies revealed pulmonary edema, hemorrhage and hyaline thrombi. Pathologic examination of the heart also showed hyaline thrombi. Information from out patients with thrombotic thrombocytopenic purpura implicates respiratory dysfunction as a component of this disease as well as the classically described pentad. Cardiogenic and noncardiogenic pulmonary edema and possibly bleeding into the lung contributed to pulmonary impairment.


Babesiosis: malaria mimic.

Babesiosis: malaria mimic. Human infection with blood parasites of the genus Babesia can result in a hemolytic disease very similar to malaria. Laboratory diagnosis of babesiosis is dependent upon the detection and identification of the parasite in blood films. The distinguishing criteria are: lack of pigment in infected red blood cells; no circulating schizonts or gametocytes; "atypical" malarialike organisms; tetrad groups, rods, or exclamation-mark forms; and persisting parasitemia after treatment for malaria.


Treatment of hyperprolactinemic amenorrhea by intermittent administration of bromocryptine (CB 154). The results obtained with intermittent bromocryptine treatment from the fifth day after the beginning of menstruation until the second day after the basal temperature rise, in 14 women with hyperprolactinemic amenorrhea-galactorrhea and in whom menses had previously been induced by continuous treatment, are presented. All women had menses without reappearance of galactorrhea; serum FSH, LH, estradiol, and progesterone followed a normal physiologic trend. Only prolactin rose again in the second phase of the cycle, in which lower levels of progesterone were also found, but without any significant interference in the clinical and hormonal trend. It is stressed that administration of bromocryptine during the first part of the cycle might substitute for continuous administration, thereby reducing drug consumption and hence possible side-effects as well as the cost of treatment.


Clinical, biochemical, and radiologic reversibility of hyperprolactinemic galactorrhea-amenorrhea and abnormal sella by thyroxine in a patient with primary hypothyroidism. A case of primary hypothyroidism associated with postpartum galactorrhea-amenorrhea is reported. The configuration of the sella was compatible with pituitary adenoma, but prolactin dynamics did not support an autonomous secretory state. Replacement therapy with thyroxine led to complete reversal of clinical, radiologic, and biochemical abnormalities.


Therapeutic ultrasound in the production of ocular lesions. After testing high intensity focused ultrasound on ocular tissues in animals, to obtain damage threshold equations, we used ultrasound for ocular therapy in experimental animals. We developed a therapeutic system for use in conjunction with a low=energy diagnostic ultrasound visualization technique. Our system included a spherical focused transducer operated at a frequency of 9.8 MHz to produce a focal spot 9 cm from the transducer surface. This technique produced controlled ocular tissue damage similar to that produced by laser and xenon are photocoagulation. It can be used to treat any level of tissue, ocular or orbital, and does not require media clarity. Specific tissue absorption properties necessary for laser damage were not required to produce the desired damage.


Results of argon laser photocoagulation in presumed ocular histoplasmosis. We performed argon laser photocoagulation on 31 eyes of 30 patients with presumed ocular histoplasmosis. Nineteen of 25 eyes (76%) with neovascular membranes 0.25 disk diameter or further from the fovea had 6/12 (20/40) or better vision after an average follow-up period of 20 months. Twenty-two of these eyes (88%) had the same or improved visual acuity during this period. When compared to a similar group of untreated eyes, the apparent advantage of photocoagulation in preserving visual acuity was not statistically significant.


Primary retinal dysplasia transmitted as X-chromosome-linked recessive disorder. The familial occurrence of retinal dysplasia in five affected male children suggested X-chromosome-linked recessive inheritance. The clinical features were childhood onset, severe visual impairment, head posture, nystagmus, and strabismus. The ophthalmoscopic findings varied in shape and extension; they ranged from retinal folds to dysplastic tissue covering the posterior pole or gliosis with tumor-like protrusion in the vitreous. The marked variability of the retinal findings was paralleled by the visual acuity, which ranged from some vision to blindness. Electroretinograms coordinated well with ophthalmoscopic observations. Of the five mothers, who are the presumed heterozygous carriers, two showed retinal changes.


An intracaval cannulation for obtaining pure, mixed hepatic venous blood samples. A method for obtaining pure, mixed hepatic venous blood is described and evaluated in anesthetized cats. Hepatic vascular congestion does not occur with this "intracaval cannulation", however small elevations in central venous blood pressure were noted. Although these changes persisted they did not result in systemic vascular congestion, judging from the normal arterial and portal pressures and from the lack of progressive decrease in arterial blood pressure. Blood samples obtained using the intracaval cannulation were shown to contain identical levels of oxygen as those obtained using a more complex surgical preparation. Reflux of blood from the vena cava does not occur during sampling. The responsiveness of this sampling method to rapid changes in venous content was evaluated by following the changes in glucose balance caused by direct stimulation of the hepatic nerves. The responses measured were similar to those measured in a separate set of experiments obtained using blood samples from a surgically isolated hepatic venous supply.


Studies on platelets: XXVIII: acute thrombocytopenic purpura due to lidocaine (Xylocaine)-mediated antibody. Report of a case. A patient developed severe thrombocytopenia three days after the administration of lidocaine (Xylocaine) for dental extractions. Petechial bleeding and thrombocytopenia regressed promptly, probably aided by treatment with adrenocorticotropic hormone (ACTH) and prednisone. The patient is well one year after her experience and has received other drugs with impunity. The thrombocytopenia appeared related to the presence of a specific antibody, predominantly immunoglobulin M (IgM), heat labile, in the patient's plasma and serum. The antibody was lytic in action and needed complement. It probably coupled to the drug, forming an antigen-antibody complex able to fix itself to and cause the lysis of platelets. Thrombocytopenia was induced with the parenteral administration of a small dose of lidocaine to the patient. This case may represent the first recorded example of hematologic complication related to the use of lidocaine.


A comparative study of some combined treatment regimens in acute toxoplasmosis in mice. The effectiveness of five treatment regimens was compared to mice during acute and late infection with Toxoplasma gondii. Parasitic cure rates, as judged by failure of brain tissue from surviving mice to produce infection when inoculated into clean mice, were as follows: pyrimethamine + sulfamethoxypyridazine, 92%; clindamycin + sulfamethoxypyridazine, 75%; spiramycin + sulfamethoxypyridazine, 16.7%; trimethoprim + sulfamethoxypryridazine alone, 0%. Pyrimethamine + sulfamethoxypyridazine was the most effective combination against acute toxoplasmosis in mice.


Platelet aggregability after splenectomy in patients with normosplenism and hypersplenism. The relationship between platelet count and aggregability was serially evaluated after splenectomy in patients with normosplenism and hypersplenism, and the results were compared with those found in patients undergoing only upper abdominal surgery. The preoperative count and aggregability of platelets were significantly suppressed in patients with cirrhosis of the liver, idiopathic portal hypertension, and prehepatic portal obstruction. However, the platelet aggregability of these patients markedly increased in accordance with the platelet counts after splenectomy. In the patients with normal splenic function preoperatively, splenectomy caused a simultaneous increase in platelet count and aggregability. The platelet function was not always reflected by the count in the patients with idiopathic thrombocytopenic purpura. On the other hand, in the control patients undergoing only laparatomy the platelet count substantially increased two weeks after operation, but platelet aggregability did not differ from the preoperative value. The present results suggest that additional factors for thromboembolism such as hypotension, acidosis, or stagnant blood flow should be avoided during the peak period of reactive thrombocytosis after splenectomy, and that an appropriate use of anticoagulants or inhibitors of platelet aggregation is recommended if and when necessary.


Effects of starvation on the oocytes of Cyprinus carpio--a cytomorphological and histochemical study. Effects of induced starvation on the morphology of the oocytic nucleus in Cyprinus carpio have been studied to assess the nature of structural aberrations caused and the adaptations induced in the oocytes during the period of stress on account of inanition. An attempt has also been made to study the fate of a number of metabolites during this period. It has also been observed that the nuclear membrane undergoes partial or complete degeneration and there is a proliferation in the number of nucleoli which tend to move into the cytoplasm. In addition, the degenerating oocytes tend to release most of the metabolites into the interstitial tissue surrounding the oocytes.


Failure to separate human x- and y-chromosome bearing spermatozoa by Sephadex gel-filtration. Human spermatozoa has been filtered on Sephadex gel columns (G 50 fine powder). 50--70% of the spermatozoa put on the gel bed had been recovered within the filtrate, the remaining spermatozoa--not absorbed to gel particles--were found in the upper third of the column. The percentage of motile spermatozoa was higher (80--95%) in the first fraction as compared to the control samples (40--65%). This technique seems to be unsuitable for a successful separation of human x- and y-spermatozoa, an enrichment of x-spermatozoa couldn't be obtained within the first fractions of the filtrate.


Experimental anastomosis between the parenchyma of spleen and kidney. The procedure described using UV-irradiation and surgical removement of the renal capsule leads to the development of anastomoses between the spleen and the kidney. The collaterals are able to divert blood from the portal system to the surface of the kidney and then to the renal vein and the iliac veins systems. The procedure is easily performed and well tolerated.


[Adverse effect of amniofetography on fetal thyroid function (author's transl)]. Amniofetography is a valuable tool for the diagnosis of fetal malformations. We have studied the effect of radiopaques dyes, containing large amounts of iodide, on fetal thyroid function. We have performed serial determinations of serum thyrotropin and thyroxine in seven newborn infants submitted to amniofetography. This work demonstrates that amniofetography induces a transient impairment of fetal thyroid function.


Profile comparisons of physical growth for monozygotic and dizygotic twin pairs. Three different methods of analysing similarity of physical growth curves within MZ and DZ twin pairs are compared. The concordance of the curves for each pair can be measured by either a multiple regression coefficient (R2) or the square root of the sum of squared differences between corresponding deviation points (D). The latter seems to be a more appropriate measure of profile contour similarity, mainly because the distance between curves is allowed for. An intra-class correlation based on a longitudinal analysis of variance seems to be a useful measure of average profile similarity for comparing groups of individuals. The distributions of the D measures, calculated for all MZ and DZ twin pairs, show MZ pairs to have significantly smaller D-values for both height and weight, on average, than DZ pairs. The intra-class correlations also show a higher concordance in developmental profile for MZ in comparison to DZ.


Effect of heat treatment of human milk on absorption of nitrogen, fat, sodium, calcium, and phosphorus by preterm infants. Seven very low birthweight (less than 1.3 kg) preterm infants, aged between 3 and 6 weeks were fed raw, pasteurised, and boiled human milk for 3 consecutive weeks. Serial metabolic balance techniques were used to assess the absorption and retention of calcium, phosphorous, sodium, and nitrogen, and the absorption of fat. Fat absorption was reduced by approximately one-third when raw milk was heated. It is suggested that the improved fat absorption from the raw milk may be related to the preservation of milk lipases. A reduction in the amount of N retained was also noted when the infants were fed boiled milk. There were no obvious differences in the absorption of N or the absorption and retention of Ca, P, and Na between the three milks. All infants gained weight most rapidly during the week in which they were fed raw milk. The mean weight gain during this time was approximately one-third greater than that during similar periods when pasteurised or boiled milk was administered


The portoenterostomy procedure for biliary atresia: a five year experience. The portoenterostomy (Kasai) procedure in infants with biliary atresia has dramatically altered the outlook for this heretofore fatal disease. When performed on infants under three months of age, bile drainage can be achieved in a majority of the patients. Since 1972, 37 infants have been treated with this operation at our institution. Diagnostic operative cholangiography and liver biopsy are recommended if the cause of conjugated hyperbilirubinemia is presumed to be obstructive. When biliary atresia is encountered, identification of the atretic ducts with transection high in the porta hepatis is carried out. Thirty-two infants have had the portojejunostomy, while five, in whom the proximal hepatic ducts were atretic but the gall bladder and distal ducts were patent, underwent portocholecystostomy. Examination of the resected fibrous duct tissue revealed a statistically significant correlation between ductal histology and postoperative outcome. Extended bile drainage has been achieved in 26 of 37 patients. Seventeen exhibit near normal growth and development four months to five years postoperatively. Seven have died with progressive liver disease despite bile drainage. Two additional patients died, jaundice free, from unrelated causes. Despite bile drainage, progressive hepatic fibrosis has been confirmed by serial biopsies in 14 patients. This finding indicates that biliary obstruction is not the sole component in the development of biliary cirrhosis. These data suggest that extrahepatic biliary atresia is a dynamic obliterative process, which can be favorably modified in approximately 50% of the infants by early surgical treatment.


The effects of lithium on phenylethylamine behavior in rats are counteracted by monoamine oxidase A and B inhibitors. Rats given a subcutaneous injection of beta-phenylethyl-amine (PEA) (30 or 60 mg/kg) showed stereotyped behavior and autonomic effects which were suppressed by LiCl. Inhibition of Type A MAO by clorgyline or Lilly 51641 (2 or 8 mg/kg) or of Type B MAO by l-deprenyl (2 or 8 mg/kg) counteracted the suppressant effects of LiCl on PEA-behavior. The findings suggest that both Type A MAO and Type B MAO play a role in the effects of LiCl on PEA-behavior.


Primary hypothyroidism, amenorrhea, and galactorrhea. A 30-year-old woman had galactorrhea and secondary amenorrhea, along with symptoms, signs, and laboratory findings of primary hypothyroidism. Serum prolactin and thyrotropin (TSH) levels were elevated. After initiation of thyroid hormone replacement, serum TSH and prolactin levels fell progressively into the normal range. Along with these changes, the patient noted a return of normal menstrual function and the galactorrhea disappeared.


The effect of maternal dose on lead retention in suckling rats. Lead (Pb) transfer from mother to litter was investigated at the late stage of lactation after a single intrapitoneal injection of 2.0 microgram Pb/ml marked with 203Pb. After 48 hr almost 20% of the maternal dose of 203 Pb was found in the litter, and about 0.6 and 0.2 of the injected dose was found in the liver and kidneys of suckling rats, respectively. Similar whole-body retention was observed earlier in suckling rats after a 20 times lower dose of stable lead was injected intravenously into their mothers.


Structural similarities of hog cholera virus with togaviruses. Hog cholera virus grown in PK-15 cells was purified by centrifugation through a sucrose cushion followed by sucrose gradient centrifugation. Analysis of virus labeled externally with [3H]sodium borohydride on polyacrylamide gel electrophoresis revealed two glycoproteins, gp55 and gp46. A third structural polypeptide, p36, seems not to be glycosylated. The gp46 was also found in the virus-free supernatant of infected cells. It could be demonstrated by radioimmune precipitation of virus labeled with[35S]methionine that all three polypeptides are specific for hog cholera virions. Electron microscopically hog cholera virus appeared as a spherical particle with a diameter of 42 +/- 8 nm. The virus particles frequently displayed a fringe of projections with a length of about 6--8 nm. The similarities of hog cholera virus with Alphaviruses and Flaviviruses are discussed.


[Stress-reaction of the cell. Changes in carcinoma RS-1 and Jensen sarcoma cells following administration of retinol acetate, hydrocortisone and proserine]. In the cells of RS-1 carcinoma and those of Iensen's sarcoma it has been stated that acceleration and inhibition of tumor growth induced by nonspecific influence (vitamin A, hydrocortisone, proserin) are accompanied by uniform morphological changes of intracellular structures. Therefore, it is possible to suggest that the changes observed reflect a certain cell adaptation to the altering environmental conditions (cell stress-reaction). The cell stress-reaction is presented morphologically and hence, can by systematized. An attempt of such systematization is suggested.


Auditory brain stem potentials in chronic alcohol intoxication and alcohol withdrawal. Auditory brain stem evoked responses were in unrestrained rats during periods of acute and chronic alcohol intoxication, alcohol withdrawal, and recovery. Acute alcohol administration altered the auditory brain stem potentials by a prolongation of both peak latency and central conduction time, beginning with early peaks. Similar but lesser effects affecting only the latter peaks were observed during chronic alcohol intoxication. By contrast, alcohol withdrawal resulted in a decrease in the peak latencies of auditory brain stem potentials and a facilitation of central conduction time. Recovery of the auditory brain stem potentials to the normal form required at least three to four weeks. The present study provides the first quantitative data, to our knowledge, on manifestations of alcohol tolerance and withdrawal.


[Aprotinin and the cardiac, intestinal and uterine musculature. Experimental studies in vitro]. Aprotinine had no significant effect in vitro on rat and frog heart, pregnant and non-pregnant rat uterus, rat duodenum, and guinea-pig ileum. On the other hand, aspecific depression of cardiac intestinal and uterine muscle was noted when concentrations higher than those recommended or clinical use were employed.


Cross-transmission of strongyle nematodes between macropods and domestic stock. Surveys were made of the worm burdens of feral goats, possums and Kangaroo Island Wallabies from places where macropods and ungulates graze together. Sheep and rabbits were experimentally infected with Labiostrongylus eugenii, a strongyle of the Kangaroo Island Wallaby. Neither of these investigations gave any evidence to suggest that cross-transmission of strongyles between macropods and domestic stock could occur.


Electrophysiological study of Norrie's disease. An X-linked recessive trait with hearing loss. 3 patients with Norrie's diseas were examined with electrocochleography and brain-stem-evoked responses. The examination revealed a hearing loss of pure cochlear origin with no involvement of the brain stem. The disease is inherited as an X-linked recessive trait only affecting males with unaffected female carriers. The results are thus in agreement with previous investigations of the topical lesion in genetic hearing loss.


Ligand-binding propeties of the muscarinic acetylcholine receptor in mouse neuroblastoma cells. 1. Muscarinic acetylcholine receptors in a plasma-membrane fraction derived from mouse neuroblastoma clone NIE-115 bind [3-3H]quinuclidinyl benzilate according to the Law of Mass Action (Kdissociation 40 pM, h0.96). 2. Antagonist and agonist binding to the receptor was studied by displacement of [3-3H]quinuclidinyl benzilate with non-radioactive ligands. The data show good agreement with similar data obtained on rat brain and ideal smooth muscle [Birdsall & Hulme (1976) J. Neurochem. 27, 7-16] indicating that the receptor is very similar in these three tissues.


Inhibition by low concentrations of ouabain of prolactin-induced lactogenesis in rabbit mammary-gland explants. 1. Explants of mammary tissue from pseudopregnant rabbits were cultured at 37 degrees C in air for 24-48h in Medium 199 buffered with 20mm-Hepes [4-(2-hydroxyethyl)-1-piperazine-ethanesulphonic acid]. The medium contained insulin and corticosterone, or insulin, corticosterone and sheep prolactin in the presence or absence of ouabain, an inhibitor of Na(+)/K(+)-dependent adenosine triphosphatase. The responses of explants were assessed histologically, by measuring the tissue concentration of K(+), and by rates of synthesis of RNA, protein and fatty acids. The effect of ouabain on Na(+) and K(+) concentrations in slices of lactating rabbit mammary-gland tissue incubated for 1h at 37 degrees C in Krebs bicarbonate buffer was also studied. 2. Prolactin increased the concentration of K(+) in mammary explants, an effect prevented by ouabain. In slices of lactating tissue, there was a linear relationship between the log dose of ouabain (from 0.1 to 10mum) and increased Na(+) and decreased K(+) concentrations in the tissue. 3. Ouabain at concentrations up to 1mum did not affect the rate of synthesis of RNA, protein or fatty acids by explants cultured with insulin and corticosterone. By contrast, the stimulatory effect of prolactin on protein synthesis was diminished and the induction of medium-chain fatty acid synthesis by prolactin was almost abolished. RNA synthesis was unaffected. Histological examination showed no tissue damage by 1mum-ouabain. 4. Explants cultured in the presence of 2mum-ouabain for 24h retained their ability to respond to prolactin when the ouabain was removed from the culture medium. Between 24 and 48h they showed responses to prolactin of a magnitude similar to those of explants never exposed to ouabain. 5. These results show that a fully functional Na(+)/K(+)-dependent adenosine triphosphatase system is necessary for prolactin to promote secretory activity in rabbit mammary gland.


Studies on the proteins of Poecilocera picta. I-changes taking place in the haemolymph protein during moulting. The haemolymph proteins of the 6th nymph of P. picta were fractioned by the polyacrylamide gel disc electrophoresis. A total of seven proteins fractions have been detected from the haemolymph. The chemical nature of different protein fractions have been examined by histochemical methods. The changes taking place in the cuticle and epidermal cells have been examined during the transformation of 6th nymph into adult. The fat body proteins have been electrophoretically fractioned and the changes in the concentration of different protein fractions have been examined. It is suggested that the protein fraction 3 of the haemolymph is utilized in the formation of new cuticle. It is concluded by the histochemical observations that proteins of the band 3 are synthesized in the fat body.


Studies on the proteins of Poekilocerus pictus. II--changes taking place in the haemolymph protein during maturation of female. In all, 7 protein fractions have been detected in the haemolymph of Poekilocerus pictus by the polyacrylamide gel disc electrophoresis. Out of seven protein fractions, four fractions i.e.3,4,5 and 6 take major role in the vitellogenesis. These fractions are accumulated in the ovariectomised female insect. No yolk is deposited in the allatectomised female ovary. The protein fractions 4, 5 and 6 show high concentration of protein in the allatectomised female as compared to the show operated female. It is concluded that the synthesis of protein fraction 3 is controlled by the corpora allata while the fractions 4, 5 and 6 are controlled by the neurosecretory cells.


Studies on the proteins of Poekilocerus pictus. III-changes taking place in the haemolymph protein during maturation of male. In all 7 protein fractions have been detected in the haemolymph of Poekilocerus pictus by the Polyacrylamide gel disc electrophoresis. Out of seven protein fractions, four fractions i.e. 3, 4, 5 and 6 take major role in the maturation of testis. Accumulation of proteins has been observed in the castrated insect. The concentration of the protein decreases and the size of the testis follicle remains small as compared to the normal male in the allatectomised insect. It is concluded that the proteins are selectively incorporated in the testis. It also appears that the synthesis of proteins and the maturation of testis is controlled by the allatum.


Some experimental observations on the cryptonephric malpighian tubules. Imago of Tenebrio, Dermestes (Coleoptera) and Lepidopterous caterpillars Pieris and Galleria were observed for the general physiology of the cryptonephric Malpighian tubules. Injection of dyes were made. The dyes were taken only by the free portion of the Malpighian tubules. Reassociated Malpighian tubules do not take dyes from the haemoly mph. They are secretory, but not excretory. The perinephric membrane in the Lepidoptera is impermeable to the dyes.


Role of haemolymph proteins in the osmoregulation in damselfly larva. In damselfly nymphs different proteins in the haemolymph have been identified which play their role in diuretic and antidiuretic functions.


Hydroxylapatite thermal fractionation of chromatin and DNA. Chromatin and DNA from developing muscle cultures were fractionated by hydroxylapatite thermal chromatography on the basis of differential thermal stability. A thermal chromatography system was developed in which protein mediated thermal stability of chromatin DNA was maximally expressed. The resulting chromatin and DNA elution profiles were similar to thermal denaturation profiles in low ionic strength solution. Additional studies showed this system was able to detect protein stabilization of DNA in in vitro nucleohistone preparations. Although some protein remained bound to hydroxylapatite during chromatin thermal elution, it did not affect the denaturation or elution behavior of free DNA on the same column. These studies show that fragments of chromatin or DNA can be segregated on the basis of differential thermal stability by hydroxylapatite chromatography.


Attempted separation of transcriptively active and inactive chromatin by hydroxylapatite thermal chromatography. Chromatin and purified DNA were fractionated by hydroxylapatite thermal chromatography. Fractions of varying thermal stability were tested for the proportions of transcribed sequences and repetitive sequences relative to the unfractionated genome. The first 80--85% of either total chromatin or purified DNA eluted from hydroxylapatite contained the same proportion of hybridizable sequences as total DNA. The remaining 15--20% of chromatin eluting at the highest temperatures was depleted of transcribed sequences. Analysis of the 20% highest melting fraction of purified DNA showed that, while the first two-thirds of this fraction contained the same proportion of transcribed sequences as unfractionated DNA, the last third, comprising about 6% of total DNA, was depleted of active sequences. Although no major differences were detected in nonrepetitive sequence complexity of chromatin fractions, there was a correlation between relative thermal stability and repetitive sequence content in fractions of both chromatin and DNA separated by thermal chromatography. Fragments eluting at higher temperatures contained a greater proportion of repetitive sequences, as indicated by a rapidly renaturing component. Most likely, the latest eluting fractions from both chromatin and purified DNA were enriched for a nontranscribed, highly reiterated, G+C rich satellite component of the chicken genome.


On the homogeneity of 11-S acetylcholinesterase. 11-S acetylcholinesterase (acetylcholine hydrolase, EC 3.1.1.7) purified by affinity chromatography of trypsin-digested homogenates was shown to be contaminated with three other active forms of enzyme. The initial purification used an affinity column of the inhibitor, N-methylacridinium ion. Chromatography of the "affinity-pure" sample on hydroxyapatite resulted in two peaks of acetylcholinesterase activity. One peak contained only a form sedimenting at 11-S (approx. 85% of the recovered activity). The other peak consisted of a 9.5-S form, in addition to 14-S and 18-S forms. The 9.5-S form (approx. 7% of the activity) co-electrophoresed with 11-S in 6% polyacrylamide gels and co-sedimented with the same form in sucrose density gradients containing 0.1 M NaCl. The purified 11-S enzyme was shown to be homogeneous by sucrose density gradient centrifugation and electrophoresis. These results indicate that 11-S acetylcholinesterase may be unsuitable for some characterization studies due to undetected contamination by the 9.5-S form.


Hormone receptors. 7. Characteristics of insulin receptors in a new line of cloned neonatal rat hepatocytes. 1. A new line of cloned, differentiated rat hepatocytes (RL-PR-C) was evaluated for its usefulness as an in vitro system for studying the regulation of the insulin receptor. 2. Insulin rapidly reversibly and specifically bound to RL-PR-C hepatocytes. Binding of tracer 125I-labeled insulin, which was competitively inhibited by native insulin as well as by proinsulin and analogs of insulin and proinsulin in proportion to their biological activity, was not influenced by glucagon, corticotropin, or human growth hormone. Anti-insulin receptor serum from a patient with Acanthosis Nigricans Type B competed with 125I-labeled insulin for binding to cell surface sites. 3. Trypsinization destroyed insulin binding sites, but these were restored by incubation under growth conditions; a 75% restoration of binding sites was achieved by one cell population doubling. 4. RL-PR-C hepatocytes responded to insulin binding by an increase in glycogen synthesis from glucose. The insulin effect was maximal at 85 nM, but was detectable at lower, more physiological, concentrations. 5. Chronic exposure (for at least 3h) of hepatocytes to insulin (10(-10)--(10(-8) M) reduced by up to 60% the number of binding sites for insulin (down-regulation). Down-regulation was prevented by cycloheximide at concentration (10 micron) sufficient to inhibit markedly protein synthesis from tracer isoleucine. Recovery from down-regulation induced by native insulin at 10(-7 M or lower concentrations was complete by 18 h under growth conditions. 6. Although RL-PR-C hepatocytes spontaneously transform after about 90 population doublings, no significant differences between normal and transformed cells were observed in insulin binding characteristics and in interaction of cells with anti-insulin receptor serum. However, transformed cells exhibited a substantially reduced (maximum of 20%) down-regulation response to insulin. 7. RL-PR-C rat hepatocytes appear, for these reasons, to be a useful model system for studying the regulation of the insulin receptor.


Uptake and metabolism of thyroid hormones by cultured monkey hepatocarcinoma cells. Effects of potassium cyanide and dinitrophenol. Cultured monkey hepatocarcinoma cell (NCLP-6E) were used to investigate the uptake and metabolism of thyroid hormones. Intracellular accumulation was shown by the failure to acutely release hormone from cells subsequently exposed to serum proteins, and by the metabolic trnasformation of the hormones to deiodinated products and their sulfates. When hepatocarcinoma cell monolayers were studied at hormone concentrations below 10(-10) M, neither KCN nor dinitrophenol inhibited uptake. Taken together with previous findings that uptake was neither saturable nor reduced at low temperature, these results indicate that this process was not active transport. Deiodination of both the phenolic and non-phenolic rings, however, was partially inhibited by KCN but not by dinitrophenol. Sulfation of 3,3'-diiodothyronine and 3'-monoiodothyronine was strongly inhibited by both KCN and dinitrophenol. Uptake of the hormones and their metabolites was also measured in suspended hepatocarcinoma cells and compared with the uptake by normal rat hepatocytes, human fibroblasts and human lymphocytes. In these experiments 1 micrometer triidothyronine and 0.47 mM dinitrophenol were used to inhibit deiodination and sulfation, respectively. Uptake was similar in all cell types. Accumulation was highest with 3,5,3'-triiodothyronine, intermediate with other compounds having iodines in both rings, lowest with compounds iodinated in only one ring, and absent with iodothronine sulfates. These findings help to explain the relative rates of metabolism of the iodothyronines and their release from the cells.


Measurement of binding of terbium to DNA. This communication demonstrates further that terbium(III) can be used as a probe for DNA. The stoichiometry of terbium binding to DNA was measured by two new methods. In the first method, calf-thymus DNA was titrated with radioactive terbium-160, which is an isotope of the common terbium-159. The resulting DNA-terbium complex was trapped and measured on millipore filters. In the second method, a peak of UV absorption of terbium was found at 219 nm and was used to measure stocichiometry. By both methods, the stoichiometry of binding was one Tb(III) for each three available phosphate groups in DNA. Finally, a rapid method was developed using terbrium-160 to measure the amount of nucleic acid in a solution.


Terbium as a solid-state probe for RNA.

Terbium as a solid-state probe for RNA. This paper continues previous work on the analysis of nucleic acid-terbium complexes in the solid state. The fluorescence excitation and emission spectra of the RNA-terbium(III) complex is reported. The fluorescence excitation and emission spectra of both the RNA-terbium(III) and DNA-terbium(III) complexes as trapped on millipore filters is reported. One hundred percent of the DNA combined with terbium was trapped on millipore filters. Deoxyribonucleic acid was recovered from DNA-terbium(III) complexes trapped on millipore filters using SDS-extraction. Energy transfer was shown to occur from the bases in nucleic acids to the terbium ion, whereas the actual binding of terbium to nucleic acids was due to phosphate groups. The relative fluorescence of homopolyribonucleotide-terbium complexes showed that the guanine moiety was responsible for most of the observed fluorescence. Binding studies showed an equal affinity of radioactive terbium for all the homopolyribonucleotides. The fluorescence of solid-state DNA and RNA terbium complexes was used to measure picomole quantities of DNA or RNA.


General observations on the growth and development of the young pouch opossum, Didelphis virginiana. A 3-year study of general growth and development revealed a uniform increase in body length of young pouch opossums during the first 10 weeks of life. Throughout this period, growth was linear and constant for all animals regardless of sex, litter size, or whether animals were obtained from first, second or third litters. Body weights were somewhat more variable, but there were no significant sex differences. Various aspects of external gross morphology are presented and discussed as they relate to growth of the young opossum.


Demonstration of a pump-mediated efflux in the epithelial potassium active transport system of insect midgut. The larval midgut epithelium of lepidopteran insects (e.g., Hyalophora cecropia and Manduca sexta) actively transports potassium from hemolymph to lumen when mounted in a chamber. The potassium active transport is rheogenic and does not require the presence of other alkali ions. The transepithelial potential difference, short-circuit current, and electromotive force of active transport are rapidly diminished by anoxia. The efflux of potassium, opposite in direction to potassium active transport, dramatically increased in anoxia, whereas the effluxes of sodium, cesium, and chloride did not increase in anoxia. The increase in efflux was found to have an alkali selectivity similar to that of potassium active transport. It is concluded that the rise of efflux in anoxia is due to the change characteristics of the epithelial potassium active transport mechanism in anoxia.


[Study of chicken and mouse interferons by the isoelectric focusing method]. Results of investigation of the chicken and mouse interferons by electrofocussing in sucrose and in polyacrylamide gel are described. The isoelectric points of these interferons were determined. The advantages of the electrofocussing in gel over that in sucrose for the interferon analysis was shown.


[Chronobiologic aspect of the mechanism of action of lithium salts in experimental alcoholism]. Ethanol, used for 2 months in a 5% solution, disorganized the circadian cycle of the hystophysiologic activity of the rat epiphysis. During prolonged (course) administration in the doses close to those used at the clinic lithium chloride normalized the mentioned changes and also prevented their development in case of joint two-months use of ethanol and lithium salt. This effect of the drug correlates with the inhibition or prevention of the ethanol preference development observed under these perimental conditions. A possible interrelationship between the discovered capacity of lithium salt to normalize the chronobiological derangements of the epiphysis and clinical efficacy of these salts in alcoholism and other affective periodical disorders is discussed.


The locus coeruleus noradrenergic system--evidence against a role in attention, habituation, anxiety and motor activity. Lesions of the locus coeruleus system were induced by combined stereotaxic injections of 6-OH-dopamine to the ascending fibres and just lateral to the locus coeruleus itself, to deplete the noradrenaline content of both the cerebral and cerebellar cortices. A group of rats with cortical noradrenaline concentrations of less than 30 ng/g were compared with a group with lesser destruction of the system (mean noradrenaline concentration 71 +/- 44 ng/g) and with controls (mean noradrenaline 347 +/- 58 ng/g). Lesioned rats showed normal motor activity and exploration (assessed with a holeboard) and showed normal habituation of these behaviours. The lesioned rats gave no evidence of increased susceptibility to distracting auditory stimuli whilst licking for water, and the groups did not differ in their rate of habituation to these stimuli, or in dishabituation. In a social interaction test, lesioned animals showed a decrease in social contacts in an unfamiliar situation (interpreted as a response to anxiety) of similar magnitude to that seen in the control group. In this test, lesioned animals engaged in more 'aggressive' behaviour (boxing and wrestling) than did the controls. These findings are incompatible with hypotheses that the locus coeruleus system is an integral part of the physiological mechanisms which control gross motor behaviour, attention, habituation or anxiety. Together with previous findings with the benzodiazepines, the results with the social interaction test make it unlikely that the benzodiazepines exert their anxiolytic effects by inhibiting the locus coeruleus system.


Contralateral turning evoked by the intranigral microinjection of muscimol and other GABA agonists. Contraversive turning was evoked by the microinjection of GABAergic agents into the substantia nigra (SN) of the rat. Muscimol, the most potent GABA agonist, evoked contralateral turning when injected into the SN in doses of 0.005, 0.05, 0.5 and 5 microgram, whereas 0.5 microgram of muscimol applied at extranigral sites produced no turning. A shorter lived contraversive turning response was evoked by the intranigral micro-injection of imidazole acetic acid (10 or 50 microgram), ethanolamine-O-sulphate (25 or 50 microgram), or GABA (50 microgram). No increase in GABA-induced turning was produced by local pretreatment with pipecolic acid (5 microgram). When injected into the SN, neither picrotoxin, in doses of 0.1, 0.5, 1.0 or 2.0 microgram, nor bicuculline methiodide (Bm), in doses of 0.1 or 0.2 microgram, elicited a significant amount of turning. Picrotoxin, however, partially blocked the turning evoked by the intranigral injection of muscimol, both via the i.p. and intranigral routes of administration whereas Bm did not. In addition, haloperidol (1 mg/kg i.p.) antagonized the muscimol-induced turning. Hence, we feel GABA mimetic substances injected within the SN might evoke contralateral turning via activation of a heretofore undescribed neural system arising from the SN or by activating the ipsilateral dopaminergic neurons projecting from the SN.


Cell-free synthesis of cod preproinsulin. Poly(A)-containing ribonucleic acid from cod islet greatly stimulated the incorporation of radioactive amino acids into proteins when assayed in a wheat germ translation system. The translation products were examined by specific immunoprecipitation with guinea pig anti cod insulin antibodies and by extraction with acid--ethanol. These measurements revealed at least a fivefold increase in incorporation of labelled amino acid over the nonprogrammed system. Sodium dodecyl sulfate disc gel electrophoresis and gel filtration chromatography showed a product of molecular weight 12 500, a size considerably larger than cod proinsulin (9000). It is concluded that cod proinsulin is synthesized via a larger precursor, preproinsulin.


The role of temperature and light on inhibition of development of Cooperia oncophora. Infective Cooperia oncophora larvae were temperature and light conditioned by holding them at various temperatures and daily light intensities for up to 56 days prior to administration to parasite-free calves via stomach tube. The calves were killed either on day 20 or day 21 postinfection. A marked (up to 80.5%) inhibition of development occurred in larvae held at 4 degrees C while little or no inhibition occurred in larvae held at room temperature or in fresh larvae. Marked inhibition also occurred in larvae held at 15 degrees C for 56 days while a low rate of inhibition occurred in larvae held at 17 degrees C for 42 days. Low incidence of inhibition was recorded in two of four calves given larvae held at ambient temperatures of mid-summer while appreciable inhibition of development of larvae occurred in two calves permitted to graze during the second and third week of September in 1975. Temperature conditioning of C. oncophora eggs for 27 days did not result in inhibition of development of infective larvae subsequently cultured from the eggs. Photoperiod or the presence of light did not have any appreciable effect on the development of inhibition of C. oncophora under the conditions of these investigations.


Serological studies on leptospirosis in cattle in east central Alabama. Serological surveys of leptospiral antibodies in cattle were carried out in Macon and the surrounding counties of East Central Alabama. A total of 286 bovine serum samples were screened for the presence of antibodies against live antigens from twelve pathogenic leptospiral serotypes using a microscopic agglutination test. The most frequently encountered serotypes were Leptospira hardjo (47%), Leptospira wolffi (34%), Leptospira canicola (12%), Leptospira pomona (10%) and Leptospira ballum (10%). Leptospira autumnalis, Leptospira grippotyphosa, Leptospira icterohemorrhagiae, Leptospira pyrogenes and Leptospira tarassovi were observed in less than 5% of the samples.


Cholesteryl ester depletion from the ovaries of superovulated female rats fed a normal or essential fatty acid deficient diet. The cholesteryl ester content of the ovaries was determined in rats diets containing corn oil or hydrogenated coconut oil (essential fatty acid (EFA) deficient) and subjected to superovulation by injection of luteinizing hormone and follicle-stimulating hormone. Superovulation increased ovarian weight; the effect was greater in animals fed corn oil. Superovulation significantly decreased total ovarian cholesteryl ester concentration in animals fed corn oil, with disproportionately large decreases occurring in the esters of 20:1, 20:2, 22:5w6, and 22:6w3. Significant decreases were observed in these esters when the data were expressed on a unit mass of tissue basis or in relation to total ovarian mass. In superovulated, EFA-deficient rats, esters of 18:1, 20:1, 22:5w6, and 22:6w3 were significantly lower per unit mass of tissue but this was due, in all cases except that of 22:6w3, to the increased mass of ovarian tissue; there was no decrease in total esters per ovary weight during superovulation of deficient rats. The pattern and degree of selective changes in ovarian cholesteryl esters during superovulation were different from those previously reported for adrenal esters of stressed rats.


Effect of three antagonists on the development of bacterial leaf streak of rice. Among 11 epiphytic microorganisms one species each of Pseudomonas, Erwinia, and Aspergillus were antagonistic to Xanthomonas translucens subsp. oryzicola. Symptoms of bacterial leaf streak did not develop when the antagonists were sprayed on rice leaves 24 h before inoculation. Although the symptoms developed when the antagonists were applied 24 h after inoculation, the number of lesions and their length was significantly reduced over control. When the mixture of each antagonist and the pathogen was applied, no symptoms developed with Pseudomonas and Aspergillus sp. However, the symptoms could develop with Erwinia sp. although the number and length of the lesions was reduced over control.


DNA-dependent RNA polymerase from T8 Guerin tumor. DNA-dependent RNA polymerases from nuclei of T8 Guerin tumor were studied. Two enzymes were purified several hundred times by the use of ammonium sulfate precipitation, DEAE-cellulose and phosphocellulose chromatography. One of them belongs to A(I) RNA polymerases and the second to B(II) as was established from their metal and ionic strength requirements. activity in the presence of native and denatured DNA and the resistance to a-amanitin inhibition. The quantity of class A enzyme was increased compared to B, a fact observed with most neoplastic tissues so far studied. This increase of the polymerase responsible  for ribosomal RNA synthesis could probably be related to malignant transformation in animals.


Synergism between the antiproliferative activities of arabinosyladenine and N6-benzyladenosine. N6-Benzyladenosine is a competitive inhibitor of adenosine deaminase from L-1210 cells in axenic culture as well as a potent antiproliferative agent in vitro and in vivo. Potentiation of the growth inhibitory activity of 9-beta-D-arabinosyladenine (ara-A) was observed in the L-1210 system with maximum synergism with a mixture of 16 micron ara-A and 10 micron benzyladenosine. Kinetic studies with L-1210 cell lysates showed values for Km of 0.25 mM ara-A and Ki of 0.23 mM benzyladenosine. It is suggested that ara-A and benzyladenosine in suitable combination may be expected to demonstrate enhanced clinical chemotherapeutic effectiveness.


The circadian variations in the epithelial growth of the hamster cheek pouch: quantitative analysis of DNA distributions. The pronounced diurnal rhythm in DNA distributions of the hamster cheek pouch epithelium both in the S fraction and in the (G2 + M) fraction was compared with previous studies of the changes in tritiated thymidine labelling index and mitotic activity. The DNA distributions were obtained by flow cytometry after ultrasonic disaggregation of the isolated epithelium into a suspension of single nuclei. The DNA distributions were analysed with the computer program of J. Fried (1976) and by planimetry. The S fraction was higher than the autoradiographic labelling index during the whole 24 hr period. Only the computer fitted S fraction and the labelling index had the same difference between maximal and minimal values, and maxima at the same time of day. The DNA distributions showed a diurnal release of G1 cells into S phase proceeding through (G2 + M) phase and returning to G1 phase within a 24 hr period.


Nucleic acid profile of the EMT6 cell cycle in vitro. Simultaneous RNA and DNA estimations were carried out during the cell cycle of EMT6/M/CC cells growing in vitro following synchronization by mitotic selection. The determinations were performed with a flow cytofluorimeter on individual cells stained with acridine orange. It was found that the RNA content increased during G1 then remained virtually constant between early and mid S phase, but a second increase occurred during late S. The rate of uptake of tritiated uridine paralleled these changes in RNA levels, and it was also found that the rate of uptake in metaphase and anaphase was virtually zero, but a rapid increase occurred in telophase. The increase in DNA during S was approximately linear, and the intermitotic phase and cycle durations were very similar to previously reported results.


Role of microtubules in the intracellular transport of growth hormone. Pulse-chase experiments utilising (3H)leucine have been used to study the effects of colchicine and vinblastine on intracellular transport and secretion of newly synthesised growth hormone from rat anterior pituitary fragments. Growth hormone was isolated from medium and fragments by polyacrylamide gel electrophoresis. When colchicine or vinblastine, which disrupt microtubules, were added immediately after pulse labelling, inhibition of the subsequent secretion of newly synthesised growth hormone was detected throughout the succeeding 5h. Similar inhibition was seen if the drugs were added after a 1h delay. However, if colchicine or vinblastine were added only after a 2h chase incubation, then no significant effect on subsequent release of labelled growth hormone was seen. The results suggest that these agents may inhibit the transport of newly formed growth hormone storage granules from the Golgi complex to the cytoplasmic pool. Microtubules do not appear to be involved in the mechanism of the final secretion of newly synthesised hormone by exocytosis.


Filtration effect of endothelial fenestrations on chylomicron transport in neonatal rat liver sinusoids. The function of endothelial fenestrations in the control of the passage of chylomicrons from the blood to the liver parenchymal cells was studied ultrastructurally in neonatal rats. Measurement of circulating chylomicrons and endothelial fenestrations as well as chylomicrons in the space of Disse at 2--20 h after the onset of suckling revealed a substantial filtration effect of endothelial fenestrations on chylomicrons larger than 250 nm in diameter. Direct uptake of chylomicrons by parenchymal cells was observed to occur by pinocytosis; Kupffer cells endocytosed only very few chylomicrons, and endothelial cells were inactive in this respect.


Microsurgical transection of small nerve fibre bundles in vitro. Effects on axons, growth cones and glial cells. Fibres growing from neurons of explanted dorsal root ganglia from 10 day chick embryos were transected and subsequently observed by light and electron microscopy after periods of a few to fifty minutes. Changes immediately proximal and distal to the cut together with alterations further away from the site of injury on both sides of the cut were recorded. Observations were also made on the growth cones of damaged axons and on changes in associated glial cells. Reactive and degenerative changes including the rotation, retraction and swelling of cutaxons occurred rapidly. Electron microscopy revealed tracts of filamentous material close to the sealed-off ends of axons, swollen organelles such as mitochondria, and lamellar bodies of varying dimensions. Proximal to the injury and closer to the explant, damaged and degenerating axons mingled with normal processes. Many contained only a fine granular material, others clumps of organelles, particularly mitochondria. Distal to the cut, microspikes were lost from some growth cones. The dense granular material filling microspikes and growth cones remained unchanged. Clumps of large clear vesicles, lamellar bodies and swollen degenerating mitochondria were present, not only within growth cones, but also in all parts of the axon distal to the cut. Glial cells associated with transected axons soon developed an electron dense cytoplasm containing swollen organelles. Large numbers of vesicles filled with a particulate substance were also found. The possible significance of the changes observed after transection are considered and discussed.


Distribution of myosin and the glial fibrillary acidic protein (GFA protein) in rat spinal cord and in the human frontal cortex as revealed by immunofluorescence microscopy. The glial fibrillary acidic (GFA) protein and myosin were localized in rat spinal cord and human frontal cortex using specific antibodies against GFA protein from human spinal cord and highly purified smooth myosin from chicken gizzard by means of an indirect immunofluorescence microscopical approach. A strong GFA protein and myosin immunoreactivity was found in astrocytes of the white and grey matter and in the external glial limitans membrane. The very fine branches of astrocytic processes stained with anti-GFA protein, but not with anti-myosin. Similar results were obtained with the human frontal cortex, where myosin antibodies failed to reveal the very fine branches of protoplasmic astrocytes. As a whole, staining with the GFA protein antiserum was more crisp than with the myosin antibody.


Monoclonal antibodies as probes for differentiation and tumor-associated antigens: a Forssman specificity on teratocarcinoma stem cells. A set of monoclonal antibodies derived by fusing P3-NS1/1-Ag4-1 myeloma cells with spleen cells from a rat immunized with mouse spleen were screened for activity against a tumor cell panel. One of these antibodies was found to react only with mouse embryonal carcinoma cells and no other tumor cell type tested, including differentiated derivatives of teratocarcinomas. In the adult mouse, this antigen is expressed by subpopulations of cells in the spleen, bone marrow, lymph node, brain, kidney and testes, although not in liver and thymus. This antigen has a species and tissue distribution consistent with that of Forssman antigen. The molecules which carry this specificity on the embryonal carcinoma cells appear to be glycolipids.


[Ultrastructure of the red pulp of the spleen in various thrombocytopenias]. The authors describe 10 cases of samples of spleen tissue obtained from patients with various thrombocytopenias which had been subjected to electron microscopical examinations. In splenomegalies associated with liver cirrhoses a certain proportion of platelets accumulated in the spleen, whereas others were ingested by macrophages. No significant polymorhism of the platelets was observed. In cases of idiopathic thrombocytopenic purpura marked polymorphism of the platelets as regards size and structure was observed. The platelets also showed a concoisuous relation to the polymorphonuclear laucotytes. The alterations of the inner structure of the platelets could not be regarded as specific, because they were also found in lymhogranuloma.


Change in permeability of liposomes caused by methylmercury and inorganic mercury. The effects of two mercurial compounds, methylmercury and inorganic mercury, on lipids were examined by measuring permeability change of lipid bilayer, liposome. Both decrease in the cholesterol content and increase in the content of unsaturated fatty acid moieties in the lipid bilayers, augmented to susceptibility of the liposomes to the mercurial compounds. Inorganic mercury and methylmercury disrupted the lipid membrane to essentially the same extent. The influence on the permeability seems to be specific for mercury compounds. The significant increase in the permeability of some liposomal preparation noted even at the mercurial concentration of 10(-7) M strongly suggests that lipid in biomembrane could be one of the primary targets of these toxic substances.


The interaction of dianhydrogalactitol with DNA in cultured Yoshida sarcoma cells. Using alkaline sucrose gradient sedimentation centrifugation it was found that treatment of Yoshida sarcoma cells in culture for 1 h with increasing concentrations of dianhydrogalactitol (DAG) enhanced the sedimentation rate of DNA in a dose-dependent manner. There was no difference between the amount of protein which co-sedimented with DNA released from treated and untreated cells. When DNA was extracted from the cells using a p-amino-salicylate-phenol mixture, the protein content of DNA seemed not to be affected by DAG. The possibility that DAG could form interstrand cross-linking in cellular DNA was suggested from renaturation studies. The appearance of a fast sedimenting DNA in the alkaline sucrose gradient and the evidence for a cross-linked DNA detected by renaturation technique, only appeared later than 6 h after treatment. A similar delayed effect on the depression in the rate of DNA synthesis was also observed. These data suggest that the inhibition of DNA synthesis may be related to the delayed formation of DNA interstrand cross-linked.


Trisomy 8 mosaicism syndrome. Report of monozygotic twins. Monozygotic twins were born with the phenotypical appearance of the trisomy 8 syndrome. The first twin, a stillborn, had autopsy findings suggestive of trisomy 8 syndrome. Cultured lymphocytes and skin fibroblasts of the second, liveborn twin, showed trisomy 8. While the lymphocyte culture showed 46/47, +8 mosaicism, with normal cells predominating, skin fibroblasts yielded only cells with trisomy 8. Bone marrow preparation showed only normal cells, 46,XY. Repeat lymphocyte culture at age 14 months, showed a reduced number of trisomy 8 cells. We would like to emphasize the importance of cytogenetic studies in early infancy, using both peripheral lymphocyte and skin fibroblast cultures, to increase the chance of detecting chromosomal abnormalities.