Renal excretion of pseudoephedrine in the rat. Pseudoephedrine is an organic base used in the treatment of upper respiratory tract disorders. Surgical techniques and experimental procedures were developed to study the renal elimination mechanisms for this drug in the rat. The ability to measure renal clearance accurately and to demonstrate renal secretion by a carrier-mediated transport system was verified by comparing results from N'-methylnicotinamide (NMN) excretion studies with literature results. Renal tubular secretion of NMN was shown to be saturable and was inhibited by mepiperphenidol to the same extent as that reported in the literature. Pseudoephedrine was cleared by the kidney at a rate in excess of inulin and close to or possibly greater than renal plasma flow. In addition to filtration and secretion, pseudoephedrine appeared to be subject to pH dependent passive reabsorption. When the secretion of pseudoephedrine was studied in detail, it was found to be nonsaturable for plasma levels of pseudoephedrine ranging from 0.16 to 1.5 microgram/ml. Secretion, however, was inhibited by mepiperphenidol suggesting a carrier-mediated process.


Renal calcium and magnesium excretion during vasopressin administration into sheep with acid or alkaline urine. 1. The proposition that changes in renal calcium excretion during vasopressin administration are positively correlated with concurrent changes in urine hydrogen ion concentration was tested by administration of vasopressin into twelve conscious diuresing sheep receiving either alkalinizing or acidifying infusions. 2. Vasopressin-induced antidiuresis in sheep with alkaline urine was associated with significant increases in urinary pH and decreases in the rate of calcium excretion whereas antidiuresis in sheep with acid urine was associated with significant decreases in urinary pH and no consistent effect on calcium excretion. 3. Magnesium excretion increased during vasopressin administration in most experiments regardless of urinary pH changes. 4. Vasopressin administration did not significantly alter the rate of excretion of sodium, potassium, chloride and phosphate or the rates of sodium, potassium, chloride, inulin, para-aminohippurate and osmolal clearance in sheep with either acid or alkaline urine. Potassium excretion and clearance in sheep with alkaline ruine was higher than that of sheep with acid urine during vasopressin infusion. 5. The results support the hypothesis that changes in renal tubular hydrogen ion concentration or bicarbonate concentration caused by water reabsorption from the collecting duct and possibly the late distal tubule could be part of the explanation for changes in renal calcium excretion which occur during vasopressin-induced antidiuresis.


Abnormal testes in reindeer, Rangifer tarandus. In a sample of reindeer from South Georgia, 4 males were abnormal in that 1 had bilateral cryptorchid testis, 2 each had 1 cryptorchid testis and 1 had a vestigial testis. The antlers of the cryptorchid males were small, but the antler cycle itself was hardly affected.


Effect on fertility of storing fowl semen for 24 h at 5 degrees C in fluids of different pH. Improved storage of fowl semen above 0 degree C was achieved by adjusting the pH of the diluent. The fertility obtained with semen stored for 24 h at 5 degrees C in diluents buffered at different pH values was compared with that of semen stored in a basic, unbuffered solution. The most satisfactory result was achieved with diluent buffered at pH 6.8 OR 7.1. Worst fertility was obtained at pH 5.8 and pH 7.4 did not prove very satisfactory. There were indications that the effect of pH under the conditions of the experiment was to regulate metabolism and thereby influence the maintenance of the fertilizing ability of the spermatozoa.


Response to cryptorchidism of the testis and epididymis of the opossum (Didelphis virginiana). Adult male opossums, Didelphis virginiana, were rendered hemicryptorchid for 35 days. The cyrptorchid testis exhibited a significant reduction in weight, while the contralateral testis had a compensatory weight gain compared with testes of untreated animals. Histological changes in the cryptorchid testis included fibrosis of the tunica propria, involution of the seminiferous tubules and an apparent increase in the interstitial tissue. Many seminiferous tubules were empty and germinal cells were absent. Some Sertoli cells persisted, but the cytoplasm was vacuolated. Cryptorchid testes were characterized by mononuclear leucocytic invasion around the tubules, and some eosinophils were observed. Cryptorchidism in the opossum may induce a reaction similar to experimental orchitis.


Effect of sodium and potassium concentrations and pH on the maintenance of motility of rabbit and rat epididymal spermatozoa. Spermatozoa were collected from the caput and cauda epididymidis of rabbits and rats and diluted in Hank's solution containing BSA, with various concentrations of Na+ and K+. Ionic strength and osmolarity were kept constant. Motility was assessed at various intervals during incubation at 25 degrees C. In the pH range 7.05--7.20, the motility of rabbit spermatozoa was not affected by changes in the ratio of K+ to Na+. Similarly, the motility of rat cauda spermatozoa was not altered, but that of caput spermatozoa was slightly depressed by a high K+/Na+ ratio. In the pH range 5.45--5.85, rabbit cauda and caput spermatozoa had much greater motility in media with a high K+/Na+ ratio. The reverse result was obtained for the rat. These findings indicate that the motility of epididymal spermatozoa is influenced by external Na+ and K+ concentrations and that this phenomenon is pH-dependent.


The identity of the acid and alkaline phosphatases of human seminal plasma. Examination of human seminal plasma showed that there was no p-nitrophenylphosphatase activity maximum at alkaline pH values. A constant ratio of enzyme activities in 8 split ejaculates, identical temperature dependence and copurification through a three-step purification procedure led us to conclude that the alkaline phosphatase in human semen is identical to acid phosphatase.


Prostaglandin E-2-9-ketoreductase in ovarian tissues. The existence of the enzyme prostaglandin E-2-9-ketoreductase which can convert prostaglandin E-2 to prostaglandin F-2 alpha was indicated in experiments with pig and human ovarian tissues in vitro, using radioimmunoassay methods and a superfusion technique. Further studies involving radiotracer techniques demonstrated that the enzyme was localized in the high-speed (105 000 g) supernatant fraction of human, pig and rat luteal tissue and human stromal tissue. The enzyme was shown to be NADPH-dependent and its activity in luteal tissue increased in the order : pig less than human less than rat.


(2,6-Methano-3-benzazocin-11 beta-yl)alkanones. 1. Alkylalkanones: a new series of N-methyl derivatives with novel opiate activity profiles. A general stereospecific synthesis of (N-methyl-2,6-methano-3-benzazocin-11 beta-yl)alkanones is described and applied to the preparation of a series of alkyl ketones wherein the alkyl group is a straight or terminally branched chain containing from one to six carbon atoms. Several compounds with methoxy groups in the aromatic ring are in the morphine range of potency; they are uniformly inactive as phenazocine antagonists. Phenolic analogues range up to 100 times as potent as morphine. Those containing five or six carbon atoms in the alkyl group exhibit phenazocine antagonist activity, in one case equivalent to naloxone. This compound (3e) is selective for phenazocine in its antagonist action.


"Fingerprinting" beta-haemolytic streptococci by their production of and sensitivity to bacteriocine-like inhibitors. A scheme for the "fingerprinting" of streptococci according to their production of (P typing) and sensitivity to (S typing) bacteriocine-like inhibitory substances has been developed. P typing of 450 beta-haemolytic streptococci by their action on a set of nine standard indicator strains revealed that 80% of strains produced one or more detectable inhibitors, and that 17 different P types could be recognised. Production of some inhibitors seemed to be a property of strains of a particular serological group or type. Bacteriocine-like substances were produced by streptococci of serological groups, A, B, C, D, E, F and G. Nine strains were selected as standard producers for S typing. These strains differed in their spectra of inhibition, but all seemed to be active only against gram-positive bacteria. One producer, a group-F streptococcus, specifically inhibited group-A streptococci. The conditions of incubation were critical for demonstration of inhibitor production. A requirement for blood and for incubation at 32 degrees C were important factors. None of the inhibitors was induced by ultraviolet irradiation. The observed inhibitory effects were not attributable to either hydrogen peroxide or low pH, but to the production of a variety of substances having diverse physicochemical properties and production requirements. Most of the inhibitors do not seem to be produced in liquid media. The "fingerprinting" procedure is simple and inexpensive, and provides a reliable means of subdividing streptococcal strains that may find application as a supplement to the existing serological typing schemes.


Fatty metaplasia in a postpubertal undescended testis: a case report. The first case of intratesticular fatty metaplasia in a postpubertal undescended testis is presented. Its differential diagnosis and etiology are discussed briefly.


Characterization of an immune complex kinase in immunoprecipitates of avian sarcoma virus-transformed fibroblasts. Kinase activity detected in immune complexes containing the src gene product of the avian sarcoma virus has been reported. To further characterize this immune complex kinase, we developed a routine quantitative assay involving trichloroacetic acid precipitation  on filters. The enzyme reaction required either Mg2+ or Mn2+, but was inactive with Ca2+. The kinetics of the phosphorylation reaction indicated a transient enzyme activity limited by rapid substrate-dependent inactivation of the enzyme. A variety of nucleoside and deoxyribonucleoside triphosphates (dATP, ATP, GTP, CTP, dGTP, TTP, dCTP) served as phosphoryl donors. The phosphorylation of immunoglobulin G was inhibited by the presence of nucleoside diphosphates. Deoxyribonucleoside diphosphates can either stimulate or inhibit the kinase reaction depending upon the concentration used. The unusual enzymatic properties of the immune complex kinase raise the possibility that the enzyme does not function as a protein kinase in vivo, but rather belongs to a different class of kinases (nucleotide kinases) which adventitiously phosphorylates immunoglobulin G when immunoprecipitated with immune serum.


[A clinical observation of cefuroxime in pediatric field (author's transl)]. Cefuroxime, a new synthetic cephalosporin, was administered to 10 pediatric patients (6 with respiratory tract infection, 2 with urinary tract infection, 1 with sepsis of E. coli and 1 with enterocolitis). The clinical result was good and excellent in all the 10 cases. No side effect was observed in any of them.


Effects of hyperglycemia and hyperthermia on the pH, glycolysis, and respiration of the Yoshida sarcoma in vivo. Tissue (extracellular) pH (pHe) and intracellular pH (pHi) were measured together in vivo in the solid Yoshida sarcoma and normal organs (liver, gastrocnemius muscle) of noninbred Wistar rats. pHe was monitored by insertion of a miniature capillary glass electrode, and pHi was measured indirectly by equilibrium partitioning of the weak organic acid 5,5-dimethyloxazolidine-2,4-dione across the cell membrane. Under normal conditions, tumor, liver, and gastrocnemius had a similar pHe of 7.05--7.30; tumor pHi was consistently higher (7.2) than that of the normal tissues (6.8--7.1). Curative hyperthermia (42 degrees C for 1 hr) did not significantly change tumor pHe or pHi. After ip glucose injection [6 g/kg body wt; blood glucose level greater than 400 mg/100 ml (22 mmoles/liter) for 4 hr], tumor pHe decreased markedly to 6.6 within 4 hours and did not return to normal for a further 12--14 hours, whereas tumor pHi was hardly affected. No marked change was noted in pHe or pHi of the normal organs following glucose loading of the host. In tumor slices removed from hyperglycemic hosts, marked reduction of both respiration and glycolysis was observed. Hyperglycemia (4 hr) plus hyperthermia at 40 degrees C (1 hr) had a synergistic inhibitory effect on metabolism that was equivalent to heat alone at 42 degrees C, and respiration and glycolysis almost ceased after 3--4 hours. However, tumor heating at 40 degrees C in hyperglycemic hosts was not equivalent to hyperthermia at 42 degrees C: With the former treatment, tumor regression did not occur, and animal survival did not differ from that of control untreated rats. The data do not support the postulate that the effects of heat on tumor cells are mediated via low pHi or that hyperglycemia leads to a lowered pHi which sensitizes the tumor to destruction at 40 degrees C instead of 42 degrees C.


[Hormone and mediator content in the blood of the adrenals and in the peripheral blood in hypertension]. The concentration of the fractions of corticosteroids, aldosterone, catecholamines, and cyclic nucleotides (cAMP and cGMP) in blood of the adrenal veins and the activity of renin in blood of the renal veins were studied in 22 patients with stages IIA and IIB hypertensive disease. At the same  time the content of these substances in the peripheral blood was determined and compared with the level of steroid and catecholamine excretion in the daily urine. An increase in the content of free 11 OCS and F fractions in the peripheral blood and blood of the adrenal veins was revealed in all patients examined.


[Non-invasive determination of oxygen saturation with the oxygenmet pulse wave oximeter on fingers, metacarpus and wrist of infants. Comparison with the oxygen saturation calculated from the pH and pO2 of the blood gas analysis (author's transl)]. Oxygen saturation was determined non-invasively with the oxygenmet pulse wave oximeter 1471 and mathematically from the pH and pO2 of the blood gas analysis. The determination of the oxygen saturation by the aid of the oximetric method was carried out on fingers, metacarpus and wrist of 80 infants (male = 44, female = 36 - ages: 1 day - 1 year and 8 months). Blood gas analysis was done just behind the oximetric analysis and oxygen saturation was calculated from the pH and pO2. The comparison of the oxygen saturations determined non-invasively on fingers, metacarpus and wrist among themselves, showed no statistical significant differences. Also no statistical significant differences were observed between the results obtained with the pulse wave oximeter and the results calculated from the blood gas analysis. The comparison of the oxygen saturations determined with the oxygenmet oximeter (mean of fingers, metacarpus, wrist) with the oxygen saturations calculated from the blood gas analysis showed a very close correlation. The correlation coefficient was 0.930.


[Panarteritis nodosa in an 11 year old boy]. The polyarteritis nodosa is a disease which can be observed in children. Its occurence in early infancy is extremely seldom and infants who contract this disease show a remarkable rate of high mortality. In 4 cases of polyarteritis in early infancy death occured within the age of three months (8, 9, 12, 13). Patients who contract this disease have involvements of different organs. This report discribes a case of polyarteritis nodosa in an older child of 12 years of age with renal, cerebral and cardiac involvement. The histological clinical and laboratory findings are discussed.


alpha-Glucosidase in Pompe's disease.

alpha-Glucosidase in Pompe's disease. Isoelectric precipitation at pH 5.0 and the use of the inhibitors, turanose, maltose and citrate, enabled the diagnosis of Pompe's disease to be made in dextran-isolated leucocytes using 4-methylumbelliferyl-alpha-D-glucopyranoside as substrate. These techniques were unnecessary with lymphocytes as the deficiency of acid alpha-glucosidase could be shown directly.


Alcohol intake, ratio of plasma alpha-amino-n-butyric acid to leucine, and gamma-glutamyl transpeptidase in nonalcoholics. In a sample of healthy men, there appeared to be no relationship between reported drinking and the ratio of plasma alpha-amino-n-butyric acid to leucine or between this ratio and an index of liver dysfunction (gamma-glutamyl transpeptidase).


Diagnosis and management of fetal distress. Both biophysical and biochemical techniques may be used to diagnose fetal distress. Fetal heart rate monitoring should be thought of as a screening technique to define a population at significant risk for fetal acidosis. The addition of fetal scalp blood sampling improves the clinician's diagnostic accuracy. The hallmark of treatment is to alleviate the stress on the fetus, to restore intervillous and cord blood flow, and, hence, to improve fetal oxygenation. This improvement may be accomplished by (1) discontinuing oxytocin, (2) correcting maternal hypotension, (3) administering oxygen to the mother, and (4) attempting to alleviate cord compression by changing the relationship of the fetal presenting part to the umbilical cord and pelvis.


Decay pattern of rabbit erythrocyte hexokinase in cell aging. As previously reported, during rabbit red blood cell aging glucose phosphorylating activities show several modifications. In the first period of the red cell life span the predominant form is similar to hexokinase II, while in the mature erythrocyte the predominant glucose phosphorylating activity resembles hexokinase I. In the oldest cells glucose phosphorylating activity has a low affinity (high Km) for glucose. In this paper the modifications of hexokinase in cell aging have been studied in vivo in a young erythrocyte population synchronized by actinomycin D, and in vitro in red cells separated in fractions according to different ages. Since protein synthesis is lacking in the mature red cell, we are inclined to explain the presence of low-affinity hexokinase activity in the oldest erythrocytes as an age-dependent transformation of a primary hexokinase.


The effect of chronic ethanol administration on central neurotransmitter mechanisms. Rats were maintained on ad lib food and a forced-intake regimen of ehtanol for up to 270 days. The ethanol treatment induced alterations in the metabolism of central catecholamines seen as increased endogenous concentrations of dopamine concomitantly with decreased concentrations of noradrenaline in the limbic forebrain. The synthesis of catecholamines, measured as the accumulation of dopa following inhibition of aromatic amino acid decarboxylase, was unchanged during chronic ethanol treatment. Local application of dopamine into the nucleus accumbens caused a greater increase in locomotor activity in chronic ethanol rats than in controls thus indicating that chronic ethanol treatment increased the sensitivity at or beyond central dopamine receptors. This phenomenon of functional dopamine receptor supersensitivity was first observed after 5 months of ethanol treatment and lasted for about 4 weeks after cessation of the ethanol treatment. The sensitivity of noradrenergic and cholinergic receptor mechanisms appeared to be unchanged after chronic exposure to ethanol. The effect of the GABAergic drug, gamma-butyrolactone (GBL) on the accumulation of dopa after inhibition of aromatic amino acid decarboxylase was studied in chronic ethanol rats. The enhancement of the dopa formation in dopaminergic neurons induced by GBL was markedly attenuated after chronic ethanol treatment. The gross behavioural depression by GBL was also weakened. This may indicate that chronic ethanol treatment causes subsensitivity of GABA receptors.


Quantitative structural analysis and the secretory behaviour of the rat parotid gland after long and short term isoprenaline treatment. Isoprenaline (IPR) was administered as daily subcutaneous injections into newborn rats for a period of 9 weeks (long-term treatment) and into 8 week-old rats for 10 days (short-term treatment). Both the parotid and the submandibular glands increased five- to six-fold in weight in the two groups due to hypoertrophy as well as to hyperplasia. The parotid glands were subjected to electron microscopic stereological analyses and to in vitro secretory studies. The results were compared with non-treated controls. Whereas mean total cell volume in the latter was 807 microns 3 it amounted to 5804 microns 3 in glands from long-term IPR-treated rats. A striking increase in size and number of cytoplasmic granules was noted after IPR treatment; there was also a marked decrease in granule electron density as compared with control cells. Granule volume density was 31.2 +/- 1.8% in controls and 58.0 +/- 1.5% in long-term IPR-treated rats. The increase in volume density, however, was accompanied by a relative decrease in amylase and cyclic AMP contents. On a percentage basis, the basal secretion of amylase from incubated IPR-treated parotid glands was markedly higher (roughly twice) than that of control glands; the absolute release of amylase into the medium, however, was only slightly increased. Basal secretion was not energy requiring, which would suggest a passive diffusion. Stimulation by beta-adrenoceptor agonists, including beta 1 and beta 2 selective agents, had no or only a small stimulatory effect in vitro on IPR-treated glands. Dibutyryl cyclic AMP was effective in controls but not in treated glands. Cholinergic stimulation caused a considerable amylase release from glands of IPR-treated rats; this release was comparable to that obtained in controls. The results suggest that superstimulation of the beta-adrenoceptor leads to a decreased sensitivity for adrenergic agonists. This may be due to membrane changes (e.g. modified receptor sites) and/or to an altered intracellular metabolism (e.g. a modified turnover of cyclic nucleotides).


Isolation and characterization of phospholipase D from fababeans. An enzyme activity in crude extract of fababeans hydrolyzed phosphatidylcholine-U-14C to produce choline and phosphatidic acid. This enzyme, phospholipase D, was stable at 50 C in the presence of 5 mM DTT but was inactivated at 55 C. The enzyme was precipitated with cold acetone, concentrated between 30% saturation to 40% saturation with ammonium sulphate, absorbed on calcium phosphate gel and eluted with 0.2 M phosphate buffer. This procedure resulted in a 20-fold increase in specific activity. The activity of fababean phospholipase D was much higher when assayed at 38 C than that at room temperature. There was an obligatory requirement for calcium, and for maximal activity 40 mM calcium was required. A narrow pH optimum of about pH 5.7 was observed. The enzyme activity was extremely dependent on substrate dispersion. When 5 mM phosphatidylcholine (PC) was sonicated with increasing levels of sodium dodecyl sulphate (1 mM to 4 mM), the enzyme activity kept increasing. By using equimolar concentrations of PC and sodium dodecyl sulphate (1 mM to 5 mM), the Michaelis constant (Km) was estimated to be 1.74 mM. Addition of choline and serine at 10 mM concentration reduced phospholipase D activity by 31% and 22%, respectively.


Divalent cation-induced aggregation of chromaffin granule membranes. Divalent cations induce the aggregation of chromaffin granule ghosts (CG membranes) at millimolar concentrations. Monovalent cations produce the same effect at 100-fold higher concentrations. The kinetics of the dimerization phase were followed by light-scattering changes observed in stopped-flow rapid mixing experiments. The rate constant for Ca2+-induced dimerization (kapp) is 0.86-1.0 x 10(9) M-1sec-1, based on the "molar" vesicle concentration. This value is close to the values predicted by theory for the case of diffusion-controlled reaction (7.02 x 10(9) M-1sec-1), indicating that there is no energy barrier to dimerization. Arrhenius plots between 10 degrees and 42 degrees C support this; the activation energy observed, +4.4 Kcal, is close to the value (4.6-4.8 Kcal) predicted for diffusion control according to theory. Artificial vesicles prepared from CG lipids were also found to have cation-induced aggregation, but the rates (values of kapp) were less than 1/100 as large as those with native CG membranes. Also, significant differences were found with respect to cation specificity. It is concluded that the slow rates are due to the low probability that the segments of membrane which approach will be matched in polar head group composition and disposition. Thus large numbers of approaches are necessary before matched segments come into aposition. The salient features of the chromaffin granule membrane aggregation mechanism are as follows: (a) In the absence of cations capable of shielding and binding, the membranes are held apart by electrostatic repulsion of their negatively charged surfaces. (b) The divalent and monovalent cation effects on aggregation are due to their ability to shield these charges, allowing a closer approach of the membrane surfaces. (c) The major determinants of the aggregation rates of CG membranes are proteins which protrude from the (phospholipid) surface of the membrane and serve as points of primary contact. Transmembrane contact between these proteins does not require full neutralization of the surface charge and surface potential arising from the negatively charged phospholipids. (d) After contact between proteins is established, the interaction between membranes can be strengthened through transmembrane hydrogen bonding of phosphatidyl ethanolamine polar head groups, divalent cation-mediated salt bridging, and segregation of phosphatidylcholine out of the region of contact.


The measurement of intracellular sodium activities in the bullfrog by means of double-barreled sodium liquid ion-exchanger microelectrodes. A double-barreled Na+-selective microelectrode was constructed with monensin as a liquid ion exchanger. The HCl-treated monensin was dissolved in a solvent (Corning 477317) at 10% (weight/weight). Internal reference solution of its ionic barrel was mixture of 0.49 M NaCl and 0.01 M KCl, the pH being adjusted to 3 with 0.1 M citrate-HCl buffer, whereas that of the PD barrel was 0.5 M KCl. Average slope and selectivity ratio (Na+/K+) tested on 10 different microelectrodes were -57.5 +/- 1.87 mV/P(Na) (SEM) and 6.7 +/- 0.44, respectively. The electrical resistance was an order of 10(10) ohm and the response time was less than 10 sec. Using this microelectrode, a free flow micropuncture experiment was carried out in the bullfrog kidney and the intracellular Na+ activity as well as the membrane PD was determined on the proximal tubular cell. Average value (+/- SEM, n = 15) for the intracellular Na+ and K+ was 20.7 +/- 1.56 mEq/L and 61.2 +/- 1.16 mEq/L, respectively, and -68.7 +/- 0.88 mV for the peritubular membrane PD. There was a significant negative correlation between Na+ and K+ activities within the cell, i.e., the lower the ionic activity of cellular Na+ was, the higher the cellular K+, and vice versa, the sum of these two being kept nearly constant. The above finding may be somehow related to the isosmosis in the reabsorptive process across the proximal tubular epithelium.


Actin from diploid and haploid cells of the myxomycete Physarum flavicomum--comparison with skeletal muscle actin. Glycerinated diploid microplasmodia of Physarum flavicomum exhibited ATP-dependent contractility typical of actomyosin controlled systems. Actin was isolated from microplasmodia, haploid amoebae-swarm cells, and rabbit skeletal muscle, by classical methods as well as ATP-DEAE cellulose chromatography, and isoelectric focusing. Actins from all three sources had isoelectric points in the pH 6.5-7.0 range, and behaved similarly during the various isolation procedures. SDS-gel electrophoresis revealed that the actin from microplasmodia and muscle had a molecule weight of about 46,000 daltons while the amoebae-swarm cells possessed proteins of molecular weights of about 46,000 and 51,000 daltons.


Plaque formation in vitro by Actinomyces viscosus in the presence of Streptococcus sanguis or Streptococcus mutans. Actinomyces viscosus, growing on a tooth in the presence of sucrose, slowly produced a loosely-attached plaque, the pH being 6.1 after 120 h. When the tooth was inoculated simultaneously with A. viscosus and either Streptococcus sanguis or Streptococcus mutans, firmly-adherent plaque was quickly formed and the pH fell below 5 after 33 h with the former Streptococcus and 24 h with the latter. A. viscosus disappeared from each mixed plaque by 120 h.


Treatment of fear-induced aggression in a horse. Desensitization (gradually exposing an animal to a fear-inducing stimulus without evoking the fear response) and counter-conditioning (rewarding the animal for behavior incompatible with the fear response) are highly successful ways of eliminating or reducing fear responses and corresponding aggression.


Incorporation of inositol into intact red blood cells. II. Enhancement of gas transport in inositol hexaphosphate-loaded red blood cells. The gas-transport function of red blood cells which have incorporated inositol hexaphosphate is significantly improved by fusion with effector-loaded lipid vesicles. "Right-shifts" of the O2-binding curves of inositol hexaphosphate-loaded red blood cells with half-saturation pressures at 37 degrees C up to 98 mmHg are observed. The transformation of 56% of the intracellular haemoglobin into the low affinity state corresponds to an optimum increase of the O2-release capacity of 270%. The CO2 transport is also correspondingly enhanced. The treated red blood cells show no inositol hexaphosphate depletion during a reasonable time.


[TEA-resistant outward current in the somatic membrane of perfused nerve cells]. The outward currents remaining after addition of 20-50 mM tetraethylammonium (TEA) to the extracellular solution were studied on perfused isolated neurons from Helix pomatia. A potassium-carried noninactivating outward current with potential-dependence and kinetics different from those of TEA-sensitive potassium currents was found. This TEA-resistant current includes a component depending on the presence of the inward calcium current. It could be abolished by replacing extracellular calcium by magnesium ions, by blocking the calcium channels with extracellular cadmium ions and their distruction by intracellular fluorid ions. An increase in the level of intracellular free carcium (by perfusing the cell with solutions containing Ca-EGTA buffer) potentiated the TEA-resistant component of the outward current and the removal of free calcium by EGTA decreased it. A conclusion is made that the somatic membrane contains outward current channels which can be activated only when calcium ions are bound to its inner surface.


The value of lorazepam in gas encephalography: a preliminary report. Lorazepam is of value in the performance of gas encephalography. Given with judicious and individualized doses of IV narcotic, it has significantly lessened the morbidity of this examination. We have been particularly impressed with the lack of unpleasant memories for the procedure. The quality of the examination was good through the patients had to be reminded, because of the amnestic effects of the drug, to hold still throughout the 6-s exposure needed for polytomography.


[Use of Ro5-4200 in bronchoscopy].

[Use of Ro5-4200 in bronchoscopy]. A simple technique employing i.v. Ro5-4200 and a local anaesthetic to obtain general anaesthesia prior to bronchoscopy, particularly after chest operations, is described. The optimal sedation offered by Ro5-4200 enables this trying examination to be performed without risk and with the best possible degree of comfort for the patient and the operator.


[Effect of cold exposure in the early postnatal period on the hormone content and tyrosine aminotransferase activity in adult rats]. The cooling of new born rats within the first 7 days of life (15 min, +2--+4 degrees) was shown to lead to stable hormonal changes in their adult state (4 to 5 months): the content of corticosterone and insulin in blood increased twice. The short cooling of adult rats resulted in the increase of trioiodothyronine content and decrease of insulin content in blood and the increased induction of tyrosine aminotransferase in the liver. The data obtained corroborate an idea that the induction of certain biochemical processes during early postnatal period, due to the involvement in the transcription of certain genes, leads to stable changes in the spectrum of metabolic processes and reactions.


Respiratory response to hypoxia and hypercapnia after elimination of central chemosensitivity. Central respiratory drive responding to pH changes was eliminated by bilateral coagulation or cold block of area S (intermediate area) on the ventral medullary surface in 7 anaesthetized cats. Arterial pH, PCO2, and PO2 (4 cats) and the respiratory response to hypoxia and hypercapnia (6 cats) were observed before and after coagulation. After coagulation in hyperoxia the arterial pH dropped from 7.30 to 7.09, the arterial PCO2 was elevated from 4.80 kPa to 8.17 kPa (6 cats). Ventilation increased by 477 ml at a PCO2a of 6.58 kPa when PO2a was reduced from 39.5 kPa to 8.5 kPa before coagulation, after coagulation ventilation increased by 241 ml (4 cats). The peripheral chemoreceptors guaranteed spontaneous breathing even in hyperoxia. The data reveal that the loss of respiratory homeostasis by elimination of the S areas is due to the loss of central chemosensitive drive with concomitant reduction of peripheral chemoreceptor effect.


Incomplete cardiac vagal blockade with atropine in the anesthetized dog. In dogs in neurolept-anesthesia the successive administration of alpha- and beta-adrenergic blocking agents and atropine, which should cause the functional equivalent of surgical denervation of the heart, always results in a marked tachycardia. The same is observed in conscious dogs, but not during methoxyflurane anesthesia. Bilateral vagotomy and administration of hexamethonium abolish the tachycardia. These observations demonstrate the presence of a vagally mediated chronotropic effect which becomes manifest when the inhibitory vagal effect is eliminated through blockade of the muscarinic receptors with atropine.


[Enzyme complex celloconingine P10x].

[Enzyme complex celloconingine P10x]. The composition and properties of the enzyme preparation celloconingine P10x were studied. The preparation contained a number of citolytic enzymes and showed proteolytic and amylolytic activities. Optimal conditions for the enzyme action were found to be: for total citolytic and hemicellulase activities pH 5.2-6.2 and 45-55 degrees, for endo-beta-glucanase activity pH 4.2-5.2 and 58-62 degrees, for proteolytic activity pH 4.0-4.8 and 64-68 degrees, and for amylolytic activity pH 3.6-4.2 and 60-66 degrees C.


[Comparative evaluation of methods for isolating total protein from the biomass of Spirulina platensis]. The methods of extracting total protein from the biomass of Spirulina platensis are discussed. The results of studying different procedures of cell wall disruption, soluble protein extraction and precipitation are presented. The best results can be obtained using mechanical disintegration of Spirulina cells for 20 min (at a temperature not higher than 25 degrees C), soluble protein extraction with 0.4% NaOH, and its subsequent precipitation at the isoelectric point with 5% HCl.


[Biosynthesis of lipase by the mold fungus Rhizopus species, strain 3-3]. Cultivation of the microscopic fungus Rhizopus sp. str. 3-3--producer was investigated. Properties of lipase were studied. The technical preparation liparisopin G3x was obtained and examined. pH optimum of the preparation was within 6.0--7.0 and temperature optimum was 37 degrees C (olive oil used as substrate).


[Preparation and properties of acid proteinase from Aspergillus awamori]. From the culture liquid filtrate of Aspergillus awamori 78-2 a preparation of acid proteinase was isolated and its properties were described. This producer was shown to have a peculiar capacity of synthesizing under submerged conditions extracellular acid proteinase as the sole proteolytic enzyme. The preparation of acid proteinase showed a comparatively high hydrolyzing capacity. Under optimal conditions the enzyme degraded hemoglobin and albumin almost completely. The possibility of purifying and concentrating acid proteinase on the ion-exchange resin KB 51 X 2 was explored.


[Thermal inactivation of milk lipoprotein lipase]. The kinetics of thermal inactivation of cow's milk lipoprotein lipase (LPL) was studied. LPL inactivation can be described by the first order q equation. Thermodynamic parameters of LPL inactivation were calculated. In the range of physiological temperatures LPL existed as two conformers. The temperature of conformation conversion was 41.5 degrees C. Glycerol increased thermal stability of milk, whereas water dilution of milk, pH shift to the acid or alkaline zone, and glycine addition to milk decreased it. It is suggested that casein micellae stabilize milk LPL.


Evolution of enzyme structure.

Evolution of enzyme structure. Three-dimensional structures of enzymes offer evidence about their evolution. There are clear examples of divergent families (e.g. mammalian serine proteases) and convergence (e.g. chymotrypsin and subtilisin). Topological similarities in dehydrogenases may reflect an ancient divergence or merely chemical constraints on protein architectures. Further experimental evidence is desirable to back up arguments based on molecular morphology. By growing microorganisms on novel foodstuffs in a chemostat, one can focus selective pressure on a specific enzyme activity. Experiments will be described in which such pressure is focused on pentitol metabolism. Examination of the fine structure of the genes responsible for this pentitol metabolism has given clues about the volution of metabolic pathways.


The evolution of genetic diversity.

The evolution of genetic diversity. The existence within natural populations of large amounts of genetic variation in molecules and morphology presents an evolutionary problem. The 'neutralist' solution to this problem, that the variation is usually unimportant to the organism displaying it, has now lost much of its strength. Interpretations that assume widespread heterozygous advantage also face serious difficulties. A resolution is possible in terms of frequency-dependent selection by predators, parasites and competitors. The evidence for pervasive frequency-dependent selection is now very strong. It appears to follow naturally from the behaviour of predators, from the evolutionary lability of parasites, from the ecology of competition and, at the molecular level, from the phenomena of enzyme kinetics. Such selection can explain the maintenance not only of conventional polymorphism but also of continuous variation in both molecular and morphological characters. It can account for the occurrence of diversity within groups of haploid and self-fertilizing organisms, and for the evolution of differences between individuals in their systems of genetic control.


[Immobilization of glucoamylase and acid proteinase on modified silichrome using N-carbethoxy-2-ethoxy-1,2-dihydroquinoline]. Using N-carbethoxy-2-ethoxy-1,2-dihydrochinoline, glucoamylase and acid proteinase were immobilized on silochrome C-80 modified by gamma-aminopropyl triethoxy silane and then succinic anhydride. The maximum percentage of protein binding was 46.0 for glucoamylase and 35.0 for acid proteinase. Acid proteinase showed a decrease of specific activity.


Game theory and the evolution of behaviour. How far can game theory account for the evolution of contest behaviour in animals? The first qualitative prediction of the theory was that symmetric contests in which escalation is expensive should lead to mixed strategies. As yet it is hard to say how far this is borne out, because of the difficulty of distinguishing a 'mixed evolutionarily stable strategy' maintained by frequency-dependent selection from a 'pure conditional strategy'; the distinction is discussed in relation to several field studies. The second prediction was that if a contest is asymmetric (e.g. in ownership) then the asymmetry will be used as a conventional cue to settle it. This prediction has been well supported by observation. A third important issue is whether or not information about intentions is exchanged during contests. The significance of 'assessment' strategies is discussed.


Arms races between and within species.

Arms races between and within species. An adaptation in one lineage (e.g. predators) may change the selection pressure on another lineage (e.g. prey), giving rise to a counter-adaptation. If this occurs reciprocally, an unstable runaway escalation or 'arms race' may result. We discuss various factors which might give one side an advantage in an arms race. For example, a lineage under strong selection may out-evolve a weakly selected one (' the life-dinner principle'). We then classify arms races in two independent ways. They may be symmetric or asymmetric, and they may be interspecific or intraspecific. Our example of an asymmetric interspecific arms race is that between brood parasites and their hosts. The arms race concept may help to reduce the mystery of why cuckoo hosts are so good at detecting cuckoo eggs, but so bad at detecting cuckoo nestlings. The evolutionary contest between queen and worker ants over relative parental investment is a good example of an intraspecific asymmetric arms race. Such cases raise special problems because the participants share the same gene pool. Interspecific symmetric arms races are unlikely to be important, because competitors tend to diverge rather than escalate competitive adaptations. Intraspecific symmetric arms races, exemplified by adaptations for male-male competition, may underlie Cope's Rule and even the extinction of lineages. Finally we consider ways in which arms races can end. One lineage may drive the other to extinction; one may reach an optimum, thereby preventing the other from doing so; a particularly interesting possibility, exemplified by flower-bee coevolution, is that both sides may reach a mutual local optimum; lastly, arms races may have no stable and but may cycle continuously. We do not wish necessarily to suggest that all, or even most, evolutionary change results from arms races, but we do suggest that the arms race concept may help to resolve three long-standing questions in evolutionary theory.


The evolutionary genetics of sexual systems in flowering plants. Population genetic studies of the evolution of breeding systems in flowering plants are reviewed. The selective advantage of a gene's increasing the selfing rate is stressed. In the evolution of outbreeding mechanisms, some strong disadvantage to selfing must therefore be acting; it is suggested that this disadvantage is inbreeding depression. Populations with no absolute barrier to selfing, and with intermediate levels of self-fertilization, appear to be the most likely starting state for the evolution of outbreeding mechanisms. There is some evidence for inbreeding depression in such populations. The evolution of distyly and dioecy are considered in some detail. An explanation for the existence of supergenes controlling these systems is proposed. The breakdown of distyly and tristyly are also considered. The evolution of recombination rates in selfing and outcrossing species is examined briefly.


The evolution of ageing and longevity.

The evolution of ageing and longevity. Ageing is not adaptive since it reduces reproductive potential, and the argument that it evolved to provide offspring with living space is hard to sustain for most species. An alternative theory is based on the recognition that the force of natural selection declines with age, since in most environments individuals die from predation, disease or starvation. Ageing could therefore be the combined result of late-expressed deleterious genes which are beyond the reach of effective negative selection. However, this argument is circular, since the concept of 'late expression' itself implies the prior existence of adult age-related physiological processes. Organisms that do not age are essentially in a steady state in which chronologically young and old individuals are physiologically the same. In this situation the synthesis of macromolecules must be sufficiently accurate to prevent error feedback and the development of lethal 'error catastrophes'. This involves the expenditure of energy, which is required for both kinetic proof-reading and other accuracy promoting devices. It may be selectively advantageous for higher organisms to adopt an energy saving strategy of reduced accuracy in somatic cells to accelerate development and reproduction, but the consequence will be eventual deterioration and death. This 'disposable soma' theory of the evolution of ageing also proposes that a high level of accuracy is maintained in immortal germ line cells, or alternatively, that any defective germ cells are eliminated. The evolution of an increase in longevity in mammals may be due to a concomitant reduction in the rates of growth and reproduction and an increase in the accuracy of synthesis of macromolecules. The theory can be tested by measuring accuracy in germ line and somatic cells and also by comparing somatic cells from mammals with different longevities.


Comparison and adaptation.

Comparison and adaptation. It has sometimes been suggested that the term adaptation should be reserved for differences with a known genetic basis. We argue that adaptation should be defined by its effects rather than by its causes as any difference between two phenotypic traits (or trait complexes) which increases the inclusive fitness of its carrier. This definition implies that some adaptations may arise by means other than natural selection. It is particularly important to bear this in mind when behavioural traits are considered. Critics of the 'adaptationist programme' have suggested that an important objection to many adaptive explanations is that they rely on ad-hoc arguments concerning the function of previously observed differences. We suggest that this is a less important problem (because evolutionary explanations generally claim some sort of generality and are therefore testable) than the difficulties arising from confounding variables. These are more widespread and more subtle than is generally appreciated. Not all differences between organisms are directly adapted to ecological variation. The form of particular traits usually constrains the form of value that other traits can take, presenting several obstacles to attempts to relate variation in morphological or behavioural characteristics directly to environmental differences. We describe some of the repercussions of differences in body size among vertebrates and ways in which these can be allowed for. In addition, a variety of evolutionary processes can produce non-adaptive differences between organisms. One way of distinguishing between these and adaptations is to investigate adaptive trends in phylogenetically different groups of species.


The question of adaptive sex ratio in outcrossed vertebrates. Of various published theories of adaptive control of progeny sex ratio only two are plausible, a physiological theory by Trivers & Willard, and a demographic theory by Verner. The first applies to species in which sons and daughters impose different costs on parents, and in which only one or very few young are produced at once. They ought to show positive correlations in the sex of successive offspring and high sex-ratio variance among progenies. Verner's theory postulates a minimization of competition for mates in neighbourhoods subject to random fluctuation in sex ratio. Optimal progenies would exactly match the population's evolutionary equilibrium sex ratio. There would be little variance among progenies. Evidence from vertebrates is unfavourable to either theory and supports, instead, a non-adaptive model, the purely random (Mendelian) determination of sex. The apparent absence of parental control of progeny sex ratio is a serious theoretical difficulty.


The spandrels of San Marco and the Panglossian paradigm: a critique of the adaptationist programme. An adaptationist programme has dominated evolutionary thought in England and the United States during the past 40 years. It is based on faith in the power of natural selection as an optimizing agent. It proceeds by breaking an oragnism into unitary 'traits' and proposing an adaptive story for each considered separately. Trade-offs among competing selective demands exert the only brake upon perfection; non-optimality is thereby rendered as a result of adaptation as well. We criticize this approach and attempt to reassert a competing notion (long popular in continental Europe) that organisms must be analysed as integrated wholes, with Baupläne so constrained by phyletic heritage, pathways of development and general architecture that the constraints themselves become more interesting and more important in delimiting pathways of change than the selective force that may mediate change when it occurs. We fault the adaptationist programme for its failure to distinguish current utility from reasons for origin (male tyrannosaurs may have used their diminutive front legs to titillate female partners, but this will not explain why they got so small); for its unwillingness to consider alternatives to adaptive stories; for its reliance upon plausibility alone as a criterion for accepting speculative tales; and for its failure to consider adequately such competing themes as random fixation of alleles, production of non-adaptive structures by developmental correlation with selected features (allometry, pleiotropy, material compensation, mechanically forced correlation), the separability of adaptation and selection, multiple adaptive peaks, and current utility as an epiphenomenon of non-adaptive structures. We support Darwin's own pluralistic approach to identifying the agents of evolutionary change.


The Wilkins Lecture, 1979: of the local movement of animals. In 1627, William Harvey was writing notes for a treatise on the movement of animals, De motu locali animalium, which in the event he neither published nor completed. As its very existence was unknown until its publication in 1959, Harvey's contribution to the study of the physiology of muscle has been unnoticed. In this treatise he deals with the different kinds of animal locomotion and examines the part played by the various motive organs, the muscles and their component parts, and the nerves. He is concerned with the problems of the initiation movement and of the complexity and diversity of movement involved in any single action. Though he could reach no satisfactory comclusion for any of these problems, the quality of his observations and of the questions that he asks is such as to justify an examination of his position relative to the muscle physiologists of the later seventeenth century.


The lamina splendens of articular cartilage is an artefact of phase contrast microscopy. The so-called lamina splendens of articular cartilage is shown to be a characteristic of phase contrast microscopy; this technique provides no evidence for an anatomically distinct surface layer. Fresnel diffraction occurs at edges separating regions of different refractive indices. These diffraction effects, when viewed under phase contrast, lead to the appearance of a bright line along the edge.


Synaptic transmission: ion concentration changes in the synaptic cleft. Currents flowing through the postsynaptic membrane of an active synapse will tend to change the concentrations of ions in the synaptic cleft. Published experimental data are used to predict (a) the sodium and potassium concentration changes in the cleft at the frog neuromuscular junction, and (b) the sodium depletion in the cleft under a Ia synaptic bouton on a cat motoneuron. Significant concentration changes are predicted at both synapses. These changes will contribute to the time dependence of the observed current and will cause the reversal potential of the current to be time dependent. At the frog neuromuscular junction, the time course of the endplate current has been shown previously to depend on the magnitude of the current flowing (at a given potential). We attribute this to changes of the cleft ion concentration. The time dependent changes of the endplate current reversal potential that we predict for the neuromuscular junction are probably too small to be detected. This is because the effects of sodium depletion and potassium accumulation on the reversal potential almost cancel. We predict that near the reversal potential small currents of complex time course will remain, i.e. no true reversl potential exists. Such currents have previously been experimentally. At the cat Ia synapse, the synaptic current is predicted to deplete a significant fraction of the available extracellular sodium ions. Consequently, the magnitude of the synaptic current should be relatively independent of the number of postsynaptic channels activated, and of the membrane potental, as has previously been found experimentally.


Influence of experimentally induced agranularity on the synaptogenesis of serotonin nerve terminals in rat cerebellar cortex. The serotonin (5-HT) innervation of the posterior vermis was studied by high resolution radioautography in both normal and X-ray-induced agranular rat cerebella, following 3 h topical superfusion with 10(-4) M 3H-5-HT. In the normal cerebellar cortex, 5-HT axonal varicosities are scarce and only rarely exhibit the membrane differentiations characterizing synaptic contacts. In the agranular cerebellum, 5-HT terminals appear to have a much higher density than in normal controls, although their absolute number may not be significantly different when the important reduction in volume of this experimental cerebellum is taken into account. These terminals frequently show typical synaptic contracts. Most of them are established on the branchlet spines of Purkinje cell dendrites, but some are also observed on the shafts of Golgi cell dendrites. The 5-HT innervation of the cerebellar cortex thus undergoes important changes in the absence of granule cells. It is suggested that these modifications may be part of the general reorganization process of the cerebellar circuitry consequent on the early destruction of the external granular layer. This new example of synaptic remodelling could imply that the formation of cerebellar connectivity is modulated, to a certain extent, by the local cellular environment.


Distribution and environmental synchronization of the marine insect, Halobates robustus, in the Galapagos islands. The following three aspects of the biology of the marine insect Halobates robustus were studied, during a two week observation period, at several sites in the Galapagos Islands: distribution, aggregation behaviour and rhythmicity of locomotory activity. H. robustus occurred in highest numbers on the water surface at shores fringed with mangroves. The aggregations of H. robustus varied according to their location and density. Copulating adults formed dense, floating aggregations, which tended to be close to rocks or mangroves. Late instar nymphs were less aggregated and, in lagoons (where there was some shelter from direct tidal forces), were furthest from the shore. In two types of habitat (mangrove-fringed, sandy shores and rocky shores) the aggregations of H. robustus showed a pronounced ability to maintain a floating station in relation to the surrounding environment, irrespective of tidal movements (in one case at 34 m from the nearest fixed objects). Evidence of the ability of the aggregations to maintain station on the water surface was also obtained by comparing the movements of H. robustus with those of floating polystyrene particles, which move passively with wind and tide. Laboratory observations and experiments indicated no clear periodicity in locomotory movements throughout a 24 h period. However, the frequency of encounters between individuals showed two daily peaks, post-dawn and pre-dusk, with fewer encounters during the day and only occasional encounters during the night. By shifting the light-dark cycle it was demonstrated that the daily bimodal rhythm of encounters is triggered by dawn and, since it is not maintained in constant light or dark, an 'hour-glass' mechanism is suggested. The contributions of single adults, of copulating pairs and of immature stages to the overall pattern of activity were also determined. Immature stages did not affect the overall rate of encounters significantly and the interactions between single and copulating pairs of adults appear to have been responsible for the bimodal pattern.


The behaviour of transporting epithelial cells. I. Computer analysis of a basic model. We analyse the non-steady state behaviour of a computer model representing functional epithelial cells. The results show that a simple model of an epithelium, containing the essential ion transport asymmetries of the original Koefoed-Johnsen-Ussing model, predicts much of the observed behaviour of 'tight-type' epithelia under various well characterized experimental conditions.


Neural tube defects in curly-tail mice. I. Incidence, expression and similarity to the human condition. The incidence of neurovertebral defects in mutant mice of the curly-tail strain was investigated and found to be similar to that observed in the same mice twenty-five years ago. The results of breeding experiments support the hypothesis of Grüneberg that the defects in these mice are probably caused by a recessive gene, the expression of which is markedly affected by the genetic background. Selection against the curly-tail phenotype for six generations did not affect the incidence of abnormalities. A marked excess of females was found among exencephalic mice, as among humans with neural tube defects. Similarly, polyhydramnios, hydrocephaly, high levels of amniotic fluid alphafoetoprotein and distinctive, rapidly adhering cells in the amniotic fluid also occurred in these mice, as in humans. The curly-tail mice thus provide a useful model for the investigation of neural tube defects in man.


Neural tube defects in curly-tail mice. II. Effect of maternal administration of vitamin A. Vitamin A, a known teratogen of the central nervous system, was administered in various doses, at the time of active neural tube closure, to pregnant curly-tail mice which have a genetic predisposition to neural tube defects (n.t.d.), and to A Strong mice, which are not so predisposed. The curly-tail mice showed an enhanced susceptibility to the teratogenic effect of vitamin A given on day 8 of gestation, demonstrating a clear gene-environment interaction. There was a differential response by the two sexes. Females seemed to be more affected by the vitamin A than males. When vitamin A was administered on day 9, instead of day 8, of gestation, the incidence of n.t.d. decreased rather than increased. Furthermore, the number of mice affected by n.t.d. was markedly lower even than that found spontaneously in untreated curly-tail mice.


Effect of experimental hydrothorax on the cough reflex in conscious cats. The authors induced experimental hydrothorax in cats by injecting dextran into the pleural cavity under brief N2O anaesthesia. They examined the parameters of cough -- elicited by mechanical stimulation of the airway mucosa -- and blood gas and pH values under normal conditions and after the injection of 50, 100, 200 and 250 ml dextran. The tests were always performed 30 min after terminating anaesthesia, i.e. in conscious animals. The free fluid in the thorax was found, in conscious cats, to reduce the inspiratory values of cough, but to have no effect on cough expiration. This is in agreement with previous findings showing that the intensity of a cough expiration does not always depend on the intensity of the preceding cough inspiration. According to this finding, the decrease in the expiratory values of cough observed during experimental pleurisy cannot be due to the actual exudate. In cats, experimental hydrothorax in doses of 200 and 250 ml leads to respiratory insufficiency. The authors further found that, for the study of interoception in the airways of conscious cats, which requires experimental induction of pathological conditions under brief anaesthesia, nitrous oxide is a convenient anesthetic.


Cure and care of neurosis. I. Cure.

Cure and care of neurosis. I. Cure. Behavioural psychotherapy has long historical roots. Recently it has led to effective treatment for selected neuroses, including phobic, obsessive-compulsive and sexual disorders. Potent therapy has become a tool of experimental psychopathology which advances theory and practice. A pervasive principle is exposure of the patient to those stimuli which evoke his discomfort until this subsides. Level of arousal during exposure does not affect outcome. Theoretical issues are reviewed which decide when exposure will be sensitizing or habituating. Both psychoanalytical and conditioning models of neurosis are out of date, and models derived more directly from clinical experiment are becoming possible. The aetiology of phobias and rituals can be seen as failed extinction rather than enhanced acquisition. Relevant phylogenetic and biological factors are discussed. At the other extreme, well-documented faith-healing indicates huge gaps in our knowledge of psychotherapy.


Two Bass Scale factors and response to placebo and anxiolytic drugs. Two principal oblique factors are identified in the Bass Social Acquiescence Scale, a measure previously shown to correlate positively with placebo response and negatively with anxiolytic drug response. The two factors appeared very similar in separate analyses of data from samples of 941 psychiatric outpatients and 1,837 college students. Also, results are presented which indicate that one factor, tentatively labeled "traditionalism", accounted for the empirically observed relationships to placebo and drug response in two clinical trials.


[Results of bone marrow transplantation in severe aplastic anemia. Bibliographic review]. A review of the present results of bone-marrow transplantation (B.M.T.) in severe aplastic anaemia is presented. Nowadays, there is little doubt that for patients with severe aplastic anaemia the treatment of choice is the B.M.T. provided always that a suitable donor exists. If the patient is fortunate enough to have a normal identical twin, the syngeneic B.M.T. without immunosuppresive conditioning must be performed. This is usually successful, though in some patients failures can be observed due to probable immunological interference. This can be overcome by a new syngeneic B.M.T. preceded by immunosuppression. The usual type of suitable donors is an HLA-identical (including locus D) sibling (allogeneic B.M.T.). Approximately 50% of patients treated in this way can become long-term survivors. The chief complications causing mortality from the allogeneic B.M.T. are graft rejection and graft-versus-host-disease (G.V.H.D.). In order to reduce the graft rejection rate, transfusions from marrow-donor and relatives prior to the transplantation should be avoided. Other probable factor influencing the final outcome of the allogeneic B.M.T. are the interval diagnosis-transplantation, age of the patient, marrow cell dose, the difference of sex between the donor and the recipients and others. Semi-incompatible and incompatible allogeneic B.M.T. are briefly considered.


[Laboratory investigation of hemoglobinopathies (author's transl)]. The importance of the haemoglobinopathies, mainly thalassaemia, as a public health problem has been emphasized. The establishment of methods for detection and diagnosis of haemoglobinopathies is the major step in laboratories dedicated to screening anemias. Tables for the diagnosis of alpha and beta thalassaemia are proposed, the methods to identify abnormal haemoglobins are discussed and a map with the relative mobilities of 115 abnormal haemoglobins by electrophoresis in alkaline buffer is shown.


The inhibitory effect of bovine rumen fluid on Salmonella typhimurium. The possible fate of Salmonella typhimurium in the rumen was investigated by monitoring rumen volatile fatty acids (VFA), lactate concentrations and pH over periods which included regular feeding and 48 h starvation. Preparations were made containing 50 per cent rumen fluid from the cow or VFA solutions, and then inoculated with S typhimurium. Viable counts before and after incubation for 24 h at 37 degrees C were compared. Incubation in broths with high concentrations of VFA and low pH resulted in a marked decrease in salmonella numbers, while lower VFA concentrations had little or no inhibitory effect on growth.


The growth of Treponema hyodysenteriae and other porcine intestinal spirochaetes in a liquid medium. A new simple method for the preparation of a liquid medium containing rabbit serum for the propagation of Treponema hyodysenteriae and other porcine intestinal spirochaetes is described. The medium, when dispensed in shallow layers and sealed under 10 per cent CO2 in nitrogen, had a redox potential not greater than -125mV and an initial pH of about 6.9 when buffered with bicarbonate. Growth of T hyodysenteriae developed more rapidly and viable counts reached higher levels at 42 degrees C than at 37 degrees C. Viable counts increased at least 10,000-fold after two to five days' incubation, depending on the temperature. Growth could be initiated from small inocula that failed to produce colonies on blood agar. Using a 1 per cent inoculum, the medium supported the growth of two strains of T hyodysenteriae through 10 serial passages.


Conditions for in vitro haemolytic activity by Corynebacterium ovis exotoxin. A substance, concluded to be Corynebacterium ovis exotoxin on the basis of properties shared, was found to have haemolytic activity below pH 6 and red cell adhesion activity at neutral or slightly alkaline pH. An acidified, solid blood agar medium was used to demonstrate the extent of haemolysis that could be obtained at optimal pH and its inhibition by immune serum. Culture in a liquid medium giving rise to a suitably acidic pH allowed titration for haemolytic activity by the exotoxin and use of supernatant in a haemolysis-inhibition test to detect antitoxin.


Diagnostic significance of pH, lactic acid dehydrogenase, lactate and glucose in pleural fluid. The pH, lactic acid dehydrogenase, lactate and glucose of pleural fluid simultaneous with serum lactic acid dehydrogenase, blood lactate and glucose were determined in 105 pediatric patients. The patients were classified into groups according to the diagnosis. The pleural fluid of empyema cases was found to have the lowest pH with a mean value of 6.83 (p less than 0.0075). The malignant pleural fluid has a relatively low pH with a mean value of 7.32. The lactic acid dehydrogenase in pleural fluid of empyema cases has the highest mean of 1,470.68 (p less than 0.05). The lactic acid dehydrogenase was found increased in malignancy, tuberculosis and parapneumonic effusion cases. Lactate level was found extremely high in empyema cases with a mean value of 13.68. It was also found extremely high in malignancy, tuberculosis and parapneumonic effusion cases. The glucose level was markedly decreased in empyema cases. There exists correlation of pleural fluid lactate to pH, lactic acid dehydrogenase and glucose, as well as that of pH to lactic acid dehydrogenase.


Direct determination of luminal diameter changes in intrapulmonary airways. A method is described for the direct measurement of changes in luminal diameter at all levels of the airway. Using this method it was found that (i) abrupt bronchiolar collapse occurred in the preterminal and terminal bronchioles once the luminal diameter was reduced to a critical level: (ii) decreased temperatures resulted in airway narrowing which was reversed by increasing the temperature to above 25 degrees C; as a rule, airway narrowing followed a cranial to caudal direction, and higher concentration of a drug being required to close the peripheral airways; (iii) bronchodilators except Carbuterol had no effect on resting bronchial tone or on acetylcholine-induced constriction in the absence of alpha-adrenoceptor blockade; (iv) at 35 degrees C rhythmic waves (frequency 6--20/min) were observed; these waves travelled from the periphery in a cranial direction.


Recent studies of psychophysiology in schizophrenia. A general introduction is given and followed by a review of recent literature under the following subheadings: electrodermal activity, cardiovascular activity, smooth pursuit eye movement, electroencephalogram, and evoked potentials. An attempt is made to assess the clinical significance of the findings reported in each area and to indicate directions for future investigation. The feasibility of defining homogeneous subgroups in schizophrenia using psychophysiological parameters is also considered. The review concludes with the recommendation that peripheral psychophysiological studies entailing (1) comprehensive recording of brain electrical activity and (2) behavioral experimentation on variables thought to be influenced by schizophrenia (e.g., sustained attentional ability) are promising directions for future research. Relationships between behavioral and psychophysiological variables determined by such studies (and possibly subgroupings) may then become the basis for neurophysiological-neurochemical investigations of specific abnormalities underlying such relationships and subgroups.


Covert transport dysfunction in the choroid plexus as a possible cause of schizophrenia. Schizophrenia and certain forms of idiopathic mental retardation may result from covert immune complex disease of the basal lamina of the choroid plexus, a process already known to cause covert transport dysfunction in similar structures of, for example, skin, bowel, kidney, and endocrines. Plexial attack could lead to cerebrospinal fluid contamination and then, via an "open" ependyma, to neurotransmitter dysfunction in the periventricular limbic brain. The immune complex mechanism implies polygenic induction, direct or autoimmune, of immune sensitivity to exogenous agents and is thus compatible with the genetic picture in schizophrenia. Candidate agents include viral coat peptides and cereal grain glutens. The glutens are known to cause immune complex skin and bowell disease variants, and some empirical evidence links them to schizophrenia. Only newer immunofluorescence methods can detect the pathology, which is otherwise silent. Systemic lupus erythematosus provides a model since it is a genetic immune complex disease strongly associated with schizophreniform psychoses, exhibits choroid plexial immunofluorescence but no central nervous system pathology by ordinary methods, and may be triggered by viruses.


[Activity and safety of fenspiride in bronchial hyperexudation (author's transl)]. This clinical trial of fenspiride (injectable and sustained-release 80 mg tablet) was carried out on patients selected because their bronchopathy was of a hypersecretory type. Interesting results were observed in 20 out of 37 cases, but they were often delayed. This group of patients included a high proportion of patients with major hypersecretion; the efficacy of fenspiride is often evident, (in 8 out of 11 cases) but it is unpredictable individually. It can be concluded from this trial that a long term treatment with fenspiride, well tolerated, can usefully be tried in patients with predominant signs of bronchial hyperexudation.


Catecholamine-mediated arrhythmias in acute myocardial infarction. Experimental evidence and role of beta-adrenoceptor blockade. Ventricular fibrillation is a major mechanism of sudden death. The cellular link between catecholamine activity and the development of serious ventricular arrhythmias may be in the formation of cyclic adenosine monophosphate (cAMP). Cyclic AMP and agents promoting cAMP accumulation allow development of slow responses which, especially in the presence of regional ischaemia, could develop into ventricular fibrillation. The role of beta-antagonist agents in the therapy of acute myocardial infarction is analysed in relation to the hypothesis linking cAMP and ventricular fibrillation. Reasons for the limited effectiveness of anti-arrhythmic therapy with beta-antagonist agents are given.


Standard oesophageal tests and practice. The specific diagnosis and institution of correct therapy for patients with oesophageal symptoms are only possible after full, sophisticated investigation. This article describes the standard oesophageal tests employed.


Effect of ionic strength on the interaction of androgens with Sertoli cell nuclei. Only 35-50% of the label accumulated after incubation of cultured Sertoli cells with 3H-testosterone was readily extractable with 0.4 M KCl during a 1 h exposure. The degree of extractability was relatively constant over the pH range 7.0-8.5 but could be increased by prolonged (15 h) exposure. While 0.1 M KCl extracted a measurable amount of label, 0.4 M KCl was significantly more efficient. Furthermore, a higher proportion of the material extracted with 0.4 M KCl was associated with macromolecular species. After a 45 min exposure to 3H-testosterone, the nuclear fraction contained primarily labeled testosterone and its 5 alpha-reduced metabolites. The relative distribution of these metabolites between salt-resistant and readily extractable forms varied between experiments. In contrast, 3H-R1881 (17 beta-hydroxy-17-methylestra-4,9,11-trien-3-one) remained essentially intact in the nuclear fraction but also was only 35% extractable with 0.4 M KCl. In conclusion, although the quantitative aspects of salt extractability appear to depend to some extent upon the extraction conditions, it is apparent that the Sertoli cell nuclear fraction accumulates a significant amount of androgen in a form which is relatively resistant to removal with 0.4 M KCl. The biological significance of this phenomenon remains to be established.


Ecdysone 3-epimerase from the midgut of Manduca sexta (L.). Ecdysone 3-epimerase was partially purified by ammonium sulfate fractionation from the 100,000 g supernate of Manduca sexta midguts. The enzyme converts ecdysone and 20-hydroxyecdysone to their respective 3-epimers, requires NADH or NADPH and O2 for this reaction, and has the following kinetic parameters: for ecdysone, Km = 17.0 +/- 1.4 microM, Vmax = 110.6 +/- 14.6 pmol min-1 mg-1; for 20-hydroxyecdysone, Km = 47.3 +/- 7.5 microM, Vmax = 131.0 +/- 3.5 pmol min-1 mg-1: for NADPH, Km = 85.4 +/- 10.6 microM; for NADH, Km = 51.3 +/- 1.3 microM. The reaction is irreversible and can be inhibited by various ecdysteroids.


Renal functional response to the mushroom poison gyromitrin. The poison gyromitrin, found in the edible false morel Gyromitra esculenta Fr. ex Pers., caused in rats an increased diuresis in which urine was produced with a weak alkaline pH, a high excretion of sodium (530%), and potassium (210%). The observed increase lasted for about 12 h and was followed by a rentention with regard to the volume and the Na-excretion for about 72 h. On the basis of [3H] inulin excretion an increased glomerular filtration was determined followed by a decrease 12 h after application of gyromitrin. An increase of creatinine and urea in the serum could not be established during the retention phase. The diuresis as well as the excretion of sodium could be antagonized by injection of an equimolar dose of pyridoxine. The hydrazine derivative N-methyl-N-formylhydrazine (MFH), which is formed rapidly from gyromitrin by hydrolysis, was without any effect on the renal function. In order to explain the effectiveness of the relatively lipophilic gyromitrin, the non-effectiveness of the more hydrophilic hydrazine MFH and the blockade of the gyromitrin effect by pyridoxine, a mechanism involving the central nervous system is discussed.


Standardization of the alkaline elution procedure using X-ray-damaged nuclear DNA. Structural modification of DNA were induced by X-irradiation of crude hepatic nuclei at various dose ranges to standardize DNA damage evaluated by the alkaline elution technique. This quantitative assay can be used as reference for DNA damage induced by the in vivo administration of mutagens and/or carcinogens involved in the environment.


[Simultaneous recording of Ca2+ and pH when studying the processes of calcium transport in sarcoplasmic reticulum]. A continuous recording of changes in the Ca2+ concentration and pH in the suspension of the sarcoplasmic reticulum fragments is shown to be possible by means of the assembled installation and using the film C2+-selective electrode. When pH of the incubation medium changes from 6.5 to 7.8 there occurs a rapid release of the preliminarily accumulated Ca2+ from it. This effect is recorded in experiments with application of the Ca2+-selective electrode and a radioactive label.


[Effect of norepinephrine, GABA and cysteine on the complex of alkaline ribonuclease with its protein inhibitor]. Norepinephrine GABA, cystein and SH-glutathione evoke a considerable decrease in the activity of alkaline RNase in the postmitochondrial fraction of the rat brain homogenate where the complex of this enzyme with its natural cytoplasmic protein inhibitor is preliminarily disturbed by the addition of pCMB. However these substances have no effect on the activity of alkaline RNase and its cytoplasmic protein inhibitor each taken separately. Evidently, the studied preparations may favour the reduction of the native state of the inactive complex of the enzyme with its inhibitor which was preliminarily disturbed by the pCMB addition.


[Organoleptic and biochemical changes in beef during refrigeration and storage]. Studies were carried out on the changes taking place in the organoleptic properties and the pH values of m. longissimus dorsi of carcass quarters of cattle, subjected to various ways of chilling and storage. It was found that in quickly chilled beef (immediately following slaughter) the processes of ripening were very slow, and the organoleptic indices were lower as against meat that was slowly cooled. The pH values of quickly chilled beef dropped at a very slow rate, and it was not until the ninth day that they reached the range below 6.0. The organoleptic properties of quickly chilled beef, including a preliminary phase (6 hours at 18-20 degrees C) were formed rapidly, and on the 7th-11th day the subjective evaluations became the same as those of slowly cooled beef.


Epithelial cell kinetics in the descending colon of the rat. I. The effect of ischaemia-induced epithelial cell loss. Epithelial cell loss was induced in the descending colon of the rat by temporary ischaemia to investigate whether this would lead to an increase in crypt cell proliferation. Shortly after the temporary ischaemia the number of cells per crypt was markedly reduced, and it was shown that the cell loss occurred mainly from the non-proliferating upper half of the crypt. The number of cells per crypt reached control values again after 24-48 h. There was a marked increase in proliferative activity, as reflected by the labelling index after 3HTdR and by the mitotic index, with peak values at 16 and 24 h after ischaemia. After 48 h the proliferative indices were normal again. The increase in crypt cell proliferation was characterized by an increase in the labelling index as well as in the mitotic index per crypt cell position. No enlargement of the proliferative cell compartment in the crypt was observed. It is most likely then that the increase in crypt cell proliferation was brought about by a shortening of the cell cycle, since the growth fraction in the lower half of the crypt approaches 1.0. The possible implications of the present data for the control of colonic cell proliferation and colonic carcinogenesis are discussed.


Epithelial cell kinetics in the descending colon of the rat. II. The effect of experimental bypass. The influence of experimental bypass on the epithelial cell kinetics in the rat descending colon was studied. It was found that the number of cells per crypt was markedly reduced at 6 weeks after bypass. The percentage of labelled crypt cells, 1 h after 3HTdR, and the distribution of labelled cells in the crypt was normal. Also the life span of the epithelial cells was the same in control and bypassed colon. The response of crypt cell proliferation to ischaemia-induced cell loss in the bypassed descending colon was similar to the one previously described for normal descending colon. This indicates that the absence of the normal luminal contents does not result in a different response of colonic crypts to induced cell loss. Furthermore, it was found that the number of cells per crypt and the proliferative activity did not change in the transverse colon after temporary ischaemia of the bypassed descending colon. This indicates that the increase in crypt cell proliferation after ischaemia-induced cell loss is a local response.


Variation in the cell cycle time in the crypts of Lieberkühn of the mouse. The durations of the phases of the cell cycle were measured at different levels in the jejunal crypts of male Balb/c mice. A mean cell cycle time of 12.3 h was found for the whole crypt. In cell positions 1 and 2, the cell cycle time was 16.7 h, and this time steadily decreased to a value of between 10 and 11 h for cell positions above 11. It is concluded that basally situated crypt cells in the mouse are cycling relatively slowly, and that they form the functional stem cell pool for the crypt. These cells may also compose the potential stem cell pool which repopulates the crypt after death of proliferative cells.


Ultrastructure of testicular biopsy from an XX male. Ultrastructural study of testicular biopsy specimens from an XX male showed hyalinized seminiferous tubules and tubules containing only mature Sertoli cells. These cells possessed large lipid inclusions as well as microfilament bundles which were perpendicular to the basement membrane and parallel to one another. The basal lamina was thickened and composed of several parallel layers with myofibroblast layers between them. The interstitium showed nodular to diffuse Leydig cell hyperplasia. Four types of Leydig cells were found: 1) normal Leydig cells with crystals of Reinke; 2) cells with abundant microcrystalline inclusions as well as microfilaments and concentric cisternae of smooth endoplasmic reticulum; 3) vacuolated cells containing numerous large lipid droplets; 4) immature Leydig cells. The different ultrastructural abnormalities found in the Sertoli and Leydig cells might be considered as the histological expression of a tubular-interstitial dysgenesis which is reflected in the high levels of gonadotropins and low levels of testosterone.


An autoradiographic analysis of the mode of proliferation in the buccal mucosa of rats incubated up to 5 hours. This study was concerned with the course of DNA synthetic activity (3H-TdR-LI-method) in the buccal mucosa of adult male. Sprague Dawley rats over an incubation period of 5 h. Interest was focussed on the influences of different media and, in particular, on temoporary changes in the proliferative activity. 3H-LI were compared in specimens (a) kept in active (= i.e., 3H-thymidine containing) medium throughout their respective incubation period and (b) pre-incubated in inactive medium for varying, but clearly defined periods before being transferred into active medium for 30 min (actual 3H-LI). Independent of the medium used the rate of DNA synthesis was markedly lowered at 60 min, the reduction being more or less significant in the different media. This was due to a temporary inhibition of DNA synthesis, which was restored after 120-180 min. In contrast, the blockade of G2-phase and/or mitosis persisted up to the end of incubation, as indicated by the unchanged number of labelled mitotic figures after 120 min. Addition of glutamine to Mc Coy's 5A markedly enhanced the activity of 3H-thymidine incorporation, but could not prevent the temporary inhibition of DNA synthesis. The biochemical mechanisms relevant for cell proliferation have been reviewed and correlated to the present results.


Correlation of DNA distribution and cytological differentiation of human cervical carcinomas. The DNA distribution of biopsy specimens from 46 patients suffering from cervical carcinoma was analysed by flow cytometry and compared with the cytological differentiation. According to morphological criteria the carcinomas were classified as highly differentiated, moderately differentiated and poorly differentiated. The results demonstrate that highly differentiated tumours contain hyperploid cells predominantly with hyperdiploid DNA content. Hyperploid cell populations in the moderately differentiated tumours are mainly in the hyperdiploid and tetraploid regions. Poorly differentiated tumours contain hypertetraploid and aneuploid cell populations. The significance of these findings is discussed.


Morphological and functional characteristics of cells infiltrating and destroying tumor multicellular spheroids in vivo. EMT6 mammary sarcoma cells were grown in vitro as multicellular spheroids to model for the heterogeneity of microenvironments and structural changes which develop in many tumors, including micrometastases. Spheroids of 700-900 micron diameter were implanted into and recovered at different times from the peritoneal cavities of sensitized or nonsensitized allogeneic and syngeneic mice. The colony forming efficiency of spheroid tumor cells recovered at 24 and 48 h from sensitized allogeneic mice was markedly decreased as compared with those from nonsensitized allogeneic or syngeneic animals. These recovered spheroids were extensively infiltrated by both lymphocytes and macrophages, which ultrastructurally had very close membrane associations with tumor cells. Host cells recovered from spheroids exhibited cytotoxic activity in an in vitro 51Cr release assay. Thus, multicellular spheroids in vivo provide a unique experimental model to study the functional capacity of host cells within a spheroical tumor. Although lacking the stroma and the vasculature of in vivo solid tumors, this model does have many similarities to in vivo tumors and is thus suitable for studying the tumor cell-host cell interactions within the tumor microenvironment. In addition, the system offers the potential for quantitative study of the effects of treatment modalities on tumor cell-host cell interactions.


The effect of retinol (vitamin A) on human lymphocytes stimulated by phytohaemagglutinin. An ultrastructural study. Retinol (Vitamin A) has been found to inhibit the stimulation of lymphocytes by certain mitogens. The ultrastructural morphology of phytohaemagglutinin-stimulated lymphocytes exposed to retinol 10 microgram/ml and 20 microgram/ml show that disaggregation of polyribosomes and formation of autophagic vacuoles ensue in the majority of the cells. Some of the lymphocyte clumps are unaffected and continue to show normal mitosis. The changes in the affected cell are similar to those seen in a cell undergoing hormonal involution and it is postulated that the effects of the retinol may be mediated by a retinol binding protein in the susceptible cells.


Control of CFUc proliferation by selective endogenous inhibitors. The susceptibility of mouse bone marrow colony forming cells (CFUc) to three different types of proliferation inhibitors in capillary semisolid agar gel was studied. GI-3, a target specific peptide containing granulocyte fraction, T4-1, an oligospecific thymic factor of proteid nature, and the alkylating cytostatics dianhydrogalactitol (DAD) inhibit myeloid colony formation as a function of concentration. The respective MED values amount to 8, 10, and 0.002 microgram/ml. When compared with this same parameter 3H-TdR incorporation into DNA of liquid bone marrow cultures showed a single fold charge for the endogenous inhibitors (GI-3, T4-1) for the cytostatic (DAD) a 3 to 4 fold lower difference. It was demonstrated, that in competitive antagonism of GI-3 and colony stimulating factor the inhibitor prevails over CSF.


Analysis of mammary tumors for cytochemical evidence of endogenous mammary peroxidase. The purpose of this study was to determine whether or not endogenous mammary peroxidase can serve as a cytochemical marker to distinguish ovarian hormone-dependent from ovarian hormone independent mammary tumors. Spontaneous mammary tumors arising in virgin C3H and GR mice (hormone independent tumors) and hormone-dependent mammary tumors arising during pregnancy in GR mice were examined. None of these tumors contained mammary peroxidase. Mammary tumors induced in Sprague-Dawley rats with methylnitrousourea (MNU) and dimethylbenzanthracene (DMBA) were also examined. These tumors included hormone-dependent and hormone independent ones. Several of the DMBA-induced hormone-dependent tumors contained a few peroxidase-positive cells, but the hormone independent tumors were negative. All of the MNU-induced tumors examined were negative for mammary peroxidase. Twenty human breast tumors (malignant and non-malignant) removed from women at surgery, were also negative for mammary peroxidase. Our results indicate that endogenous mammary peroxidase cannot be used to distinguish hormone-dependent from hormone independent mammary tumors.


Electron microscopical studies on the genesis of white adipocytes: differentiation of immature pericytes into adipocytes in transplanted preadipose tissue. In order to clarify the relationships between perivascular cells, capillaries and fat cells, with a special reference to the origin of fat cells, we have made a light and electron microscopical study on the developing epididymal adipose tissue of newborn to 5-week-old rats, and also on the differentiating, transplanted epididymal preadipose tissue from 6-day-old rats. Development of epididymal preadipose tissue progressed rapidly 6 or 7 days after birth. The preadipose tissue on the 5th day after transplantation consisted of differentiated areas with many mature fat cells, and of undifferentiated areas in which these cells were scanty. In the differentiated areas of developing epididymal preadipose tissue, both in situ and transplanted, many fat cells seemed to develop in the area immediately adjacent to growing capillaries, but cells intermediate between perivascular cells and preadipocytes were seldom observed. However, in undifferentiated areas of transplanted tissue, we found ultrastructural evidence that immature pericytes of capillaries can differentiate into preadipocytes.


Electron microscopic studies on the small intestine of rats after mechanical intestinal obstruction. The morphologic changes of the small intestine after the mechanical obstruction were studied by light and electron microscopy. After the ligation of the upper small intestine, segments of jejunum, both proximal and distal to the site of obstruction, were removed at intervals varying from 45 min to 24 h. The essential changes were found in the epithelial cells of the tips of villi, and few morphologic differences were recognized between samples proximal and distal to the site of obstruction. The most remarkable changes in the mucosa were the pseudopodlike extension of the cytoplasm, vacuolar alteration of the villus epithelial cells, desquamation of these degenerated cells, and the dilatation of the epithelial intercellular spaces. A few epithelial cells showed hypertrophy of the smooth endoplasmic reticulum. In the submucosa, vascular stasis and edema were observed throughout the course. The mechanism of fluid loss into the intestinal lumen was discussed briefly.


Intermembrane inclusions induced by anoxia in heart and skeletal muscle mitochondria. Heart and skeletal muscle from rats of different ages were incubated in vitro in an oxygen-free medium supplied with substrates in order to investigate the effect of anoxia on muscle fine structure, particulary on the mitochondria. In skeletal muscle fibers anoxia has been found to induce changes similar to those previously described in ischemic muscles in vivo namely giant mitochondria, apparently derived by mitochondrial fusion, and intermembrane inclusions with a paracrystalline structure. The plate-like inclusions are mostly located in the intracristal spaces and are closely associated to cristal membranes even in markedly swollen mitochondria. Identical inclusions have been observed in cardiac muscle cells following anoxic injury, whereas they are never found in non-muscle cells such as endothelia, fibroblasts and nerve fibers. Cardiac and skeletal muscle fibers from newborn rats maintained in an oxygen-free medium show mitochondrial swelling but no intermembrane inclusions. The different response of mitochondria from developing vs adult striated muscle to anoxia may be due to changes during postnatal development in the quality or quantity of the protein component(s) involved in paracrystal formation.


Epidermal DNA synthesis in organ culture explants. A study of hairless mouse ear epidermis. Explants of split mouse ear were incubated in organ culture for up to 48 h, and the cell proliferation was studied by the addition of Thymidine-methyl-3-H (3HTdR) to the medium during different time periods, mainly for the first 14 h of incubation. Cultures were started at 0900, 2130 and 2300. In all cases the labelling index remained stable for 6-8 h, and then increased. The mean grain count, however, was falling and so was the epidermal DNA-specific uptake of 3HTdR. Based on the experimental results, calculations can be made of the flux of cells through S. It is concluded that the increasing LI is not due to inherent diurnal variation in cell proliferation, and is not a sign of real growth but caused instead by a complete block of the cell exit from S, probably combined with periods of an increased entrance rate into S. Other methodological factors, however, may also contribute to the increasing LI. Hence, this system is not suited for the measurement of factors that influence epidermal DNA synthesis.


[Effect of 1,4-benzodiazepine tranquilizers on the activity of the hepatocyte hydroxylating complex and glucose-6-phosphatase in white rats]. Intraperitoneal administration of diazepame and phenazepame into rats /at a dose 50 mg/kg/ within 4 days did not induce liver microsomal enzymes. After administration of chlordiazepoxide at the same dose content of cytochrome P-450 was increased and the rate of dimethylaniline demethylation was elevated. Content of protein as well as NADPH-cytochrome-c-reductase and glucose-6-phosphatase activities were increased after intraperitoneal administration of all the preparations at a dose 100 mg/kg within 4 days. Experiments on the potentiation of hexenal effect demonstrated the decrease in the time of sleep in animals, treated with chlordiazepoxide at a dose 100 mg/kg of body weight.


[Action of aortic cathepsin D on serum low-density lipoproteins]. Action of aortic cathepsin D on low density lipoproteins /LDL/ from human serum was studied by means of analytical ultracentrifugation. Cathepsin D intensively hydrolysed LDL, leading to decrease in the sedimentation coefficients of the product formed. The product of degradation of LDL possessing a sedimentation coefficient of 4.5 S /in 0,04 M KBr; 0,042 M NaCl, 2 mM EDTA, 6 mM Tris, pH 7.3 /contained apparently the minimal amount of proteins, which could still maintain the stability of partially hydrolysed LDL. When the degree of hydrolysis reached 40%, a heterogenous mixure, containing the flotating and slowly sedimenting products, was formed. These products gradually lost their solubility. The data obtained suggest that the insoluble products of the proteolysis may be formed in aortic wall some disorders of the lipoprotein metabolism.


[Comparison of the activity and properties of 5'-nucleotidase in the homogenates and plasma membranes of the normal liver, hepatomas and of the liver in mice with inoculated hepatomas of varying degrees of malignancy]. Activity of 5'-nucleotidase was significantly lower in plasmatic membranes of highly malignant hepatoma 22 as compared with the activity found in normal liver tissue. The optimal activity of the enzyme from hepatoma 22 was found at pH 8.5 with AMP as a substrate. Decrease of pH value from 8.5 to 7.4 did not affect the enzymatic activity in homogenates and plasmatic membranes in the normal liver tissue. In all the experiments activity of 5'-nucleotidase was lower towards CMP as compared with AMP. The additive effect of the both substrates was observed only in experiments with hepatoma 22.


[Sleep problems and their treatment in psychosis (author's transl)]. Sleep disturbances in psychoses can mean hypo- as well as hypersomnia. In 90% of endogenous depressed patients sleep disturbances were seen, mostly as hyposomnia. In the group of schizophrenic psychotic patients only 30% had sleep disturbances. With polygraphical investigations in endogenous depressed patients a shortening of REM-latency and a disturbed sleep profile, in schizophrenic psychoses a shortened REM-rebound and a reduced amount of stages 3 and 4 were found. The treatment of choice for depressions are antidepressive drugs and sleep deprivation, for schizophrenic psychoses neuroleptic drugs. This treatments improved subjective and objective sleep disturbances with psychopathological remission at the same time. So far, only hypothetical considerations do exist about the relationship between psychopathology and sleep disturbances. It is suspected that etiological relations exist between depression and desynchronization of central sleep mechanisms and between schizophrenia and special disturbances of REM-sleep and stage 3 and 4.