Changes in plasma membrane enzyme activities during liver regeneration in the rat. Changes in activities of plasma membrane enzymes during liver regeneration may be related to the maintenance of hepatic function or to the regulation of cell proliferation. Plasma membranes were isolated from rat livers at various times after partial hepatectomy, and the specific activities of alkaline phosphatase, (Na+ + K+)-ATPase, leucine aminopeptidase, 5'-nucleotidase, and adenylate cyclase (basal and with glucagon or epinephrine) were measured. Alkaline phosphatase and (Na+ + K+)-ATPase activity increased 3.6-fold and 2-fold respectively, during the first 48 h after partial hepatectomy. The time of onset and duration of change suggest that these increases in activity are involved in the maintenance of bile secretion. Decreases in leucine aminopeptidase activity at 48--108 h and in 5'-nucleotidase activity at 12--24 h were observed, which may be involved in the restoration of protein and accumulation of RNA. The basal activity of adenylate cyclase increased after partial hepatectomy. The response of adenylate cyclase to epinephrine showed a transitory increase between 36 and 108 h after surgery, while the response to glucagon was decreased by approximately 50% at all time points through 324 h after surgery. These changes in the hormone responsiveness of adenylate cyclase are similar to those previously observed in fetal and preneoplastic liver.


A simple method for the determination of affinity and binding site concentration in receptor binding studies. In ligand binding studies, it is often difficult to apply kinetic analyses because of an uncertainty in experimental data obtained at high ligand concentrations. Under such circumstances, Kd value (an index of the affinity) and the binding site concentration may be estimated more accurately from the binding of a fixed concentration of labelled ligand observed in the presence of various concentrations of the non-labelled ligand, if the fraction of both labelled and non-labelled ligand bound is small. When there is no cooperative effect of the ligand binding, the Kd value may be calculated by subtracting the concentration of the labelled drug from the concentration of the non-labelled drug to cause a 50% reduction of the saturable binding of the labelled drug. From above values, the binding site concentration may be calculated. The proposed method is capable of examining the cooperativity of the ligand binding, the labelled drug concentration and the specific radioactivity of the labelled drug and does not require large amounts of the labelled drug.


An inducible acetate transport system in Neurospora crassa conidia. Neurospora crassa conidia possess an active transport system for the uptake of acetate. This system was characterized as: (a) energy dependent; (b) taking place against a concentration gradient; (c) saturating at higher substrate concentrations and (d) competitively inhibited by propionate. Activity of the acetate transport system can be further enhanced by preincubating conidia in 1 mM acetate medium for 180 min (the  inducible transport system). The conidial system and the inducible system have similar properties. The development of the inducible transport was dependent on RNA and protein synthesis. A genetic control of this system was further confirmed by isolating a mutant acp-i acetate permease, inducible) that fails to develop the inducible transport system.


Electron microscopic visualization of the arrangement of the two protein components of (Na+ + K+)-ATPase. The information obtained by electron microscopic examination of highly purified membrane preparations of (Na+ + K+)-ATPase after freeze-fracturing or negative staining suggests the following conclusions. The catalytic 100 000 dalton protein component penetrates with its greater 'globular' mass the plasma membrane and protudes with its smaller mass from the protoplasmic surface by a stalked knob carrying the catalytic centre. The 40 000 dalton glycoprotein component is anchored in the membrane interior by a non-pom the outer membrane surface forming a surface coat of ill-definable substructure.


Enhancement of (Ca2+ + Mg2+)-ATPase activity of human erythrocyte membranes by hemolysis in isosmotic imidazole buffer. I. General properties of variously prepared membranes and the mechanism of the isosmotic imidazole effect. 1. Membranes prepared from human erythrocytes hemolyzed in isosmotic (310 imosM) imidazole buffer, pH 7.4, show enhanced and stabilized (Ca2+ + Mg2+)-ATPase activity compared with membranes prepared from erythrocytes hemolyzed in hypotonic (20 imosM) phosphate or imidazole buffer, pH 7.4. 2. Exposure of intact erythrocytes or well-washed erythrocyte membranes to isosmotic imidazole does not cause enhanced (Ca2+ + Mg2+)-ATPase activity. 3. Exposure of erythrocyte membranes, in the presence of isosmotic imidazole, to the supernatant of erythrocyte hemolysis or to a partially purified endogenous (Ca2+ + Mg2+)-ATPase activator, promotes enhanced (Ca2+ + Mg2+)-ATPase activity. Under appropriate conditions, NaCl can be shown to substitute for imidazole. The results demonstrate that imidazole does not act directly on the erythrocyte membrane but rather by promoting interaction between an endogenous (Ca2+ + Mg2+)-ATPase activator and the erythrocyte membrane.


Enhancement of (Ca2+ + Mg2+)-ATPase activity of human erythrocyte membranes by hemolysis in isosmotic imidazole buffer. II. Dependence on calcium and a cytoplasmic activator. 1. Activity of the (Ca2+ + Mg2+)-ATPase of erythrocyte membrane may be enhanced by a cytoplasmic protein activator. The presence of Ca2+ is necessary for the ionic strength-dependent interaction between the erythrocyte membrane and the activator. This is true no matter the purity of activator (unfractionated hemolysis supernatant or partially purified activator) or the major source of ionic strength (imidazole or NaCl). 2. When the endogenous activator enhances (Ca2+ + Mg2+)-ATPase activity of the erythrocyte membrane, there is a physical association between activator and membrane. This association is not disrupted by a decrease in ionic strength to 0.005 but is reversed by exposure to 5 mM ethyleneglycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid. 3. Activator binding necessary for enhancement of (Ca2+ + Mg2+)-ATPase activity may occur during preparation of membranes or during incubation for assay of ATPase.


Is there a plasma-membrane-located anion-sensitive ATPase? II. Further studies on rabbit kidney. A study has been made to determine whether renal plasma membranes contain an HCO3 stimulated, ouabain insensitive Mg ATPase. Purified mitochondrial, microsomal and brush border membrane fractions have been isolated from rabbit kidney. The microsomal anion-sensitive ATPase activity appears to be entirely of mitochondrial origin on the basis of the effects of inhibitors of mitochondrial Mg ATPase. The brush border membrane fraction is contaminated with mitochondrial fragments and contains an Mg ATPase activity with low anion-sensitivity. Further purification of this fraction causes parallel decreases in anion-sensitivity of the Mg ATPase activity and in cytochrome c oxidase activity. These results indicate that conclusions previously reached by other investigators for a role of anion-sensitive Mg ATPase in the bicarbonate reabsorption of the proximal tubule may no longer be tenable.


The formation of phosphoenzyme of sarcoplasmic reticulum. Requirement for membrane-bound Ca2+. Membrane-bound Ca or Mg of sarcoplasmic reticulum fragments were removed by treating the membrane with EDTA or an acidic solution, and the changes in the enzymatic activities of sarcoplasmic reticulum fragments induced by these treatments were examined. With the decrease in the amount of membrane-bound Ca below 1-3-10(-8) mol/mg protein, it was demonstrated that the activity of (Ca2+ + Mg2+)-ATPase transiently increased and then diminished, that the Ca-uptake and phosphoenzyme formation declined gradually, and that the activity of Mg2+-ATPase was affected to a less extent. Sodium dodecyl sulfate-gel electrophoretic patterns of peptides from the metal-deficient membranes were the same as those of the untreated material. The level of the phosphoenzyme formation of the metal-deficient membrane was restored by increasing the amount of membrane-bound Ca, but not by increasing the amount of membrane-bound Mg.


Substrate inhibition of the (Na+, K+)-ATPase in the presence of excess Mg2+. 1. High concentrations of ATP inhibit completely the activity of (Na+, K+)-ATPase (ATP phosphohydrolase, EC 3.6.1.3) prepared from sheep brain. 2. The inhibition depends on the concentration of total ATP, i.e. complexed ATP+ free ATP. 3. The inhibition by high ATP concentrations persists in the absence of K+, and is then independent of the Na+ concentration between 2 and 140 mM Na+. 4. Raising the K+ concentration at 20 mM Na+ increases the ATP concentration required for the maximal hydrolysis rate. 5. The Hill number for the inhibition process is about three. 6. The inhibition by ATP is temperature-dependent, in that as the temperature is increased, higher ATP concentrations are required for inhibition.


Enzymatic release of iron from sideramines in fungi. NADH:sideramine oxidoreductase in Neurospora crassa. Young mycelia of the fungus Neurospora crassa contain a soluble NADH-linked sideramine reductase, which may be responsible for liberating iron in vivo from accumulated sideramines during iron-deficient cultivation. The enzymes can be assayed using a soluble supernatant fraction, EDTA, and an atmosphere of pure nitrogen. The enzyme is stable without loss of activity up to 45 degrees C and has an optimum of activity at pH 7.0. Besides coprogen (Km = 100 micrometer,  V=2.8 nmol/min per mg protein), some other ferrichrome-type compounds are reduced. However, ferrichrome, ferrirubin coprogen B and ferrioxamine are poor substrates. When the mucelia were grown in a medium containing 10(-5) M ferri iron, the activity of the reductase was found to be only 30% of that found under low iron conditions. The enzyme is inhibited by oxygen, SH-alkylating agents and partly by some detergents. Unlike the reductase of N. crassa, the corresponding enzyme from Aspergillus fumigatus revealed low reduction of coprogen and high reduction of ferrichrome, indicating genusdependent specificities of sideramine reduction enzymes in fungi. The participation of acids of the citric acid cycle as natural iron acceptors during strong iron deficiency is studied and confirmed by iron uptake measurements on isolated mitochondria.


[The functional coupling between Ca2+-ATPase and creatine phosphokinase in heart muscle sarcoplasmic reticulum]. The functional role of creatine phosphokinase (CPK) in the process of energy supply for the Ca2+-ATPase reaction and ion transport across the membrane of heart sarcoplasmic reticulum (SR) has been studied. It has been shown that isolated and purified preparations of heart SR contain significant activity of CPK. The localization of CPK on the membrane of SR has been revealed also by an electron microscopic histochemical method. Under conditions of the Ca+-ATPase reaction in the presence of creatine phosphate the release of creatine into the reaction medium is observed, the rate of the latter process being dependent upon the MgATP concentration in accordance with the kinetic parameters of the Ca2+-ATPase reaction. CPK localized on the SR membrane is able to maintain higher rate of calcium uptake by SR vesicles, as compared to that with added ATP-regenerating system. The results obtained demonstrate the close functional coupling between CPK and Ca2+-ATPase in the membrane of SR.


Spectinomycin hydrochloride in the treatment of gonorrhoea: Its effect on associated Chlamydia trachomatis infections. Sixty-three heterosexual men were successfully treated with a single injection of spectinomycin hydrochloride 2 g for urethral infections with Neisseria gonorrhoeae. Chlamydia trachomatis was recovered from the urethra of 11 of these men both before and after treatment. In six men, the organism was isolated after but not before treatment. No isolates were obtained from the remaining men either before or after treatment. All 17 of the men who yielded C. trachomatis developed post-gonococcal urethritis. Eight of 46 men from whom no isolate was obtained in their cultures developed post-gonococcal urethritis. Seventeen of 50 women successfully treated with spectinomycin for cervical infections with N. gonorrhoeae yielded isolates of C. trachomatis both before and after treatment. The organism was isolated from five women before but not after treatment, and from four women after but not before treatment. In 24 women culture for C. trachomatis was negative both before and after treatment. Spectinomycin hydrochloride in the dosage used rarely eliminated C. trachomatis from the genital tract of either men or women; in this respect it resembled two other drugs commonly used for the treatment of gonorrhoea-pencillin and ampicillin.


Effects of verapamil on (Na+ + K+)-ATPase, Ca2+-ATPase, and adenylate cyclase activity in a membrane fraction from rat and guinea pig ventricular muscle. The electrophysiologic properties and the negative inotropic effect of verapamil are most likely due to the inhibition of calcium movement across the sarcolemmal membrane. A possible biochemical basis for this inhibition of calcium movement was studied in a membrane fraction rich in (Na+ + K+)-ATPase (EC 3.6.1.3) and adenylate cyclase (EC 4.6.1.1) activity and which demonstrated Ca2+-ATPase (EC 3.6.1.3) activity. Since each of these enzymes has the potential for influencing transsarcolemmal calcium movements, the effect of verapamil on their activities was studied in this membrane fraction isolated from rat and guinea pig hearts. Ca2+-ATPase activity in the rat was 37.7 mumol Pi/mg per hour compared with 13.8 +/- 2.9 in the guinea pig (p less than 0.01). Corresponding values for (Na+ + k+)-atpase activites were 7.9 +/- 0.9 mumol Pi/mg per hour versus 10.2 +/- 1.4. Adenylate cyclase activity in the rat was 240 +/- 8 pmol/mg per minute compared with 299 +/- 27. It was found that verapamil in concentrations of 0.01-100 mg/litre (2.1 X 10(-8) to 2.1 X 10(-4) M) had no effect on the activity of the above enzymes in either species and it was concluded that a biochemical basis for the effect of verapamil on calcium flux has yet to be defined.


Effect of osmolality on renal medullary Na-K-ATPase activity in the postobstructive kidney. The aim of this study was to determine the effect of changes in osmolality on the reduced renal medullary Na-K-ATPase (EC 3.6.1.3) activity of the postobstructive kidney. The effect of osmolality on renal medullary Na-K-ATPase activity was studied by incubating tissue slices from sham-operated and bilaterally obstructed rats in media with osmolality varied before enzyme isolation using sodium chloride, choline chloride, or sucrose. Both sham-operated and bilaterally obstructed rat renal medullary enzyme showed a similar increase in activity with increased osmolality due to sodium chloride. Medullary Na-K-ATPase from the postobstructive kidney also showed increased activity with osmotic changes induced by choline chloride or sucrose. It is proposed that the decrease of Na-K-ATPase activity observed after bilateral ureteral obstruction is due, at least in part, to the loss of the solute concentration gradient in the kidney.


The effect of ouabain on the guinea pig ileum longitudinal smooth muscle: 1. ATPase activities in a sarcolemma-enriched fraction prepared with the aid of divalent cation depletion of the intact muscle. Depletion of divalent cations before fractionation of the longitudinal muscle of the guinea pig ileum yielded a sarcolemma-enriched microsomal fraction free of mitochondria. A major portion of the ATPase activity in the presence of Mg, Na, and K was due to stimulation by Na alone. A further small stimulation by K was demonstrated only in the presence of an activating factor from the 105 000 X g supernatant. Ouabain inhibited only the K activation and had no effect on the Na-stimulated Mg-ATPase.


The effect of ouabain on the guinea pig ileum longitudinal smooth muscle: 2. Intracellular levels of Ca, Na, K, and Mg during the ouabain response and the dependence of the response on extracellular Ca. Isotonic Tris-HCl containing 10 mM LaCl3 at 4 degrees C effectively removed extracellular ions in 30 min while preventing loss of intracellular ions. Intracellular Ca and Na increased during the contraction in the presence of 10 mM ouabain and then decreased during relaxation. Intracellular Na increased again during the latter part of the relaxation phase when K loss became apparent. Mg levels remained essentially constant. Ouabain responses were rapidly lost in Ca-free medium indicating that they were dependent on extracellular Ca. A 5.5-fold increase in the normal levels of extracellular K did not reduce the contraction to a submaximal dose of ouabain. A full phasic response to high K (60 mM) was observed after a 10-min exposure of the tissue to ouabain, at which time the ouabain response had returned to basal tension. The contraction to ouabain appears to be dissociated from inhibition of the Na,K-ATPase at the K site. The changes in intracellular ions indicated that ouabain contracted the muscle by increasing the plasma membrane permeability to Ca and Na and later decreased the K and Na concentration gradients, probably by inhibition of the Na,K-ATPase.


Uptake and efflux of adenine and its derivatives in Neurospora crassa. Conidia of wild-type Neurospora crassa, preincubated for 3 1/2 h in growth medium, showed a typical triphasic pattern of adenine uptake. The three phases consisted of a quick initial uptake, followed by a plateau phase, and then by a resume lowered uptake. A study of the relative influx and efflux of [14C] adenine showed that the plateau phase in fact is a period of transmembrane movement of adenine and adenine metabolites. The efflux during the plateau phase essentially cancelled out all the influx during the same period. The uptake curve derived after taking into account the effluxed portion of radioactivity indicated that the second phase represents a period of lowered uptake activity. The beginning of the lowered uptake activity during the second phase is correlated with the presence of a high intracellular level of ATP derived from exogenous [14C]adenine. At the end of the secod phase, the intracellular level of ATP is much smaller and the rate of adenine uptake increases again. Analysis of the acid-soluble pool after feeding [14C]adenine indicated the presence of other 14C-nucleotides, but no detectable levels of bases and nucleosides were present. However, chromatographic analysis of the medium indicated that efflux results essentially in the accumulation of bases. The significance of this finding in relation to efflux is discussed.


Hydrogen sulfide poisoning: review of 5 years' experience. The workforce of Alberta, a province rich in fossil fuel, faces an increasing risk of exposure to hydrogen sulfide (H2S). Basic knowledge of the population exposed during the years 1969 to 1973 inclusive was accumulated to identify the immediate medical and management problems. Data were recorded from three sources of records: the Workers' Compensation Board of Alberta, the Alberta Hospital Services Commission and the provincial coroner's office. There were 221 cases of exposure to H2S. The overall mortality was 6%; 5% of victims were dead on arrival at hospital. Admission to hospital was required for 65% of the victims arriving at a hospital emergency room (78% of the 221). Acute problems were coma, dysequilibrium and respiratory insufficiency with pulmonary edema. Increased attention to cardiopulmonary resuscitation at the exposure site and during transportation to hospital is necessary to reduce the mortality from H2S exposure. No long-term adverse effects were apparent in the survivors.


The long term effects of radiation of T and B lymphocytes in the peripheral blood after regional irradiation. The long term alterations of T and B lymphocytes in the peripheral blood of patients treated with regional irradiation for various malignancies were examined. Eighty patients were tested at various intervals after the completion of irradiation. Absolute lymphocyte counts, the percentages of T cells and B cells, and the blastogenic response to phocyte reaction (MLR) were determined. Nearly all patients initially had absolute lymphocytopenia and one-third of the patients tested 3 years after completion of irradiation had lymphocyte counts which were more than two standard deviations below the normal range. The depression was not specific for either the T-or B-lymphocyte subpopulations. The PHA response was impaired for extended periods of time after the completion or irradiation. Differences in the mean response of lymphocytes to PHA were noted for all concentrations of the mitogen, but were most marked with suboptimal concentrations of PHA. The MLR was below the lower limits of normal in 70% of the recently irradiated patients. There was a gradual recovery of the ability to respond in the MLR, and all patients tested more than 4.5 years after the completion of therapy had a normal response. These results were compared with those obtained in patients treated with total lymphoid irradiation for Hodgkin's disease. Although three appeared to be a difference in the effect of radiation on lymphocyte subpopulations in the two groups, the effects on lymphocyte function were similar.


Isolation and purification of normal and malignant colonic plasma membranes. Plasma membrane fractions from normal colon cells and a transplantable colon adenocarcinoma were isolated and purified by differential and zonal density centrifugation. Enrichment of normal and adenocarcinoma plasma membranes was found in zonal fractions I and II (ZI and ZII) following centrifugation in an 18--50% sucrose gradient. The distribution of various marker enzymes in normal colon preparations suggested an apical origin for the membranes obtained in zonal fraction I while zonal fraction II appeared to contain basal-lateral membrane fragments. Enzymatic analysis of the plasma membrane derived from the colon tumor indicated that these fractions possess a more uniform distribution of Na-K+ ATPase perhaps reflecting a dedifferentiated state. The plasma membrane fractions isolated should prove useful for investigation of transport and other properties of vesicles derived from malignant and normal colon cells.


Chemical and biological properties of B16 murine melanoma cells grown in defined medium containing bovine serum albumin. The addition of 1 percent (w/v) bovine serum albumin (BSA) to the F12 medium utilized for the growth of the B16 melanoma cells significantly stimulated the growth of this cell line. The synthesis of mucopolysaccharides and sialoglycopeptides in this medium is identical with that in Eagle's minimal essential medium with Earle's balanced salt solution supplemented with 2 mM L-glutamine, twice the recommended concentration of vitamins, nonessential amino acids, sodium pyruvate, and 10 percent (v/v) fetal calf serum. Cell volume and morphology did not change significantly, under the different growth conditions and tumorigenicity, as assayed by injection of cultured cells into syngeneic animals, was not decreased. Analysis of the BSA used indicated the presence of a sialoglycoprotein contaminant. This sialoglycoprotein contaminant was present in all lots examined and contains N-acetyl-and N-glycolylneuraminic acid, mannose, galactose, and glucosamine. The sialoglycoprotein can be removed by chromatography on acetate form anion-exchange resin at pH 4.3. F12 media containing the purified BSA plus selenite and the sodium salts of palmitic, oleic, and linoleic acids supported growth of the melanoma cells to the same extent as did the media containing unpurified BSA, indicating that the sialoglycoprotein has no role in sustaining the growth of the cells.


Effect of methotrexate and 5-fluorodeoxyuridine on ribonucleotide reductase activity in mammalian cells. A number of studies in bacteria have indicated that deoxythymidine 5'-triphosphate may be a repressor or corepressor of ribonucleotide reductase. For determination of whether a similar regulating mechanism exists in mammalian cells, HeLa cells and partially hepatectomized rats were treated with either methotrexate, 5-fluorouracil, or 5-fluorodeoxyuridine in order to block thymidylate synthesis and consequently lower the intracellular pools of deoxythymidine 5'-triphosphate. In HeLa cells there was a significant (360 to 400 percent) increase in reductase activity in both the methotrexate and 5-fluorodeoxyuridine-treated cells. The administration of methotrexate to partially hepatectomized rats resulted in a 2.7-fold enhancement of the hepatectomy-induced increase in reductase activity, and the 5-fluorouracil treatment yielded a 60 percent increment in the increase of ribonucleotide reductase activity after partial hepatectomy. Cycloheximide prevented the increase in reductase activity after the exposure of HeLa cells to methotrexate and 5-fluorodeoxyuridine, indicating that the stimulation of ribonucleotide reductase activity was the result of enhancement of de novo enzyme synthesis rather than of enzyme activation. The data support the thesis that deoxythymidine 5'-triphosphate or a thymidylate metabolite may be involved in the regulation of ribonucleotide reductase levels in mammalian cells.


Inhibition of surface immunoglobulin centeral capping of Daudi cells and cell spreading of HeLa-S3 cells by neocarzinostatin. The effects of an antitumor antibiotic, neocarzinostatin (NCS), on the surface immunoglobulin central capping induced by anti-immunoglobulin M antibody on Daudi cells and on the cell spreading of trypsinized HeLa-S3 cells were examined. Pretreatment of Daudi cells and HeLa-S3 cells with NCS, 5 to 30 micrograms/ml, for 4 hr inhibited cap formation and cell spreading, respectively. It was shown that there is a direct relationship between the dose and the degree of inhibition. Inhibitors of DNA synthesis such as bleomycin, mitomycin C, and 1-beta-D-arabinofuranosylcytosine showed no inhibitory effect on cap formation or cell spreading. However, known microtubule-acting agents such as colchicine and vinblastine sulfate completely inhibited both capping and cell spreading at a dose of 10 micrograms/ml. In view of the fact that 10 micorograms NCS per ml also inhibit the formation of microtubular paracrystals induced by vinblastine sulfate in HeLa-S3 cells and that other agents known to influence microtubule function such as local anesthetics and calcium ionophores also inhibit both paracrystal formation and cap formation, these new observations add further support to our hypothesis that NCS affects microtubular proteins transmembranously in vivo.


Inhibition of lymphocyte transformation by disrupted murine oncornavirus. Freeze-thaw preparations of banded Rauscher murine leukemia virus markedly suppressed the in vitro cellular-mediated blastogenic response of murine splenic lymphocytes to phytohemagglutinin-P and to allogeneic cells in two-way mixed-leukocyte reaction. Suppression was shown not to be due to cytotoxicity or to virus-mitogen binding. It is suggested that a virion envelope component interferes with cellular-mediated immunity by altering cell recognition sites.


Regulation of maturation rate of mouse granulocytes. Regenerating mouse bone marrow cells were cultured i.p. in diffusion chambers (DC) to study factors affecting the maturation rate of granulocyte precursors. One day after exposing 3-day-old DC cultures to 3H-thymidine the cultures were harvested, and labelled proliferative and non-proliferative granulocytes were counted in radioautographs. The relative maturation rate--defined as the fraction of proliferative precursors maturing into the non-proliferative compartment per unit time--could be increased by different experimental procedures that inhibit cell production. Inhibition was obtained (a) by increasing culture cellularity; (b) by implanting DC into normal rats or rats with huge s.c. chloroma tumours rather than into mice; and (c) by treating the cells with leucocyte extracts  (granulocyte chalone) during the last day of culture. Furthermore, a sudden inhibition of rapidly proliferating granulocytes by leucocyte extracts resulted in an increase (apparently transient) in the absolute number of labelled non-proliferative granulocytes. Such an increae was not detected in experiments involving a stronger or sustained inhibition of granulopoiesis, evidently because the size of the precursor population had been markedly reduced.


Leptofibrils in intrafusal muscle fibres of muscle spindles in the domestic fowl. Leptofibrils consisting of narrow dark and wide light bands at regular periods are commonly found in intrafusal muscle fibres of chicken muscle spindles. They are particularly abundant in intrafusal muscle fibres with the loose type of myofilaments. They occur either at the periphery of intrafusal muscle fibres or in deeper regions, or even close to sensory nerve terminals. Dark bands of some peripheral leptofibrils vary considerably in size and appear less regular in configuration. Lateral extensions from the dark bands may occur with or without interconnections. Lateral attachments to myofilaments at the immediate neighbourhood may also occur.


[Our experience in the treatment of inguinal hernia using a silastic-dacron prosthesis]. The Authors report their experience on 50 patients suffering from inguinal hernia and treated with a plastic surgery operation preceded by subfascial introduction of a Silastic-Dacron prothesis. They describe the reported complications and relate on the frequency of relapses within a period of 3 years, comparing them with the data reported by the existing literature on this subject.


The influence of venous return on cardiac mechanical and sarcoplasmic reticulum function during endotoxemia. E. coli endotoxin (0.03 mg/ml) added to blood perfusing a heart-lung preparation with a venous return of 600 ml/min produced a significant depression in ventricular function within 4 hours. Fragmented sarcoplasmic reticulum isolated from the myocardium of the endotoxin-perfused heart-lung preparations showed depressed calcium uptake rates and ATPase activity. When venous return was increased to 1,200 ml/min, gram-negative endotoxin had no effect on ventricular function, isolated fragmented sarcoplasmic reticulum calcium uptake, or ATPase activity. These observations suggest that gram-negative endotoxin or a product thereof acts in synergism with low venous return in order to depress myocardial function.


Functional morphology of the pressure- and the volume-hypertrophied rat heart. We studied hearts in which hypertrophy was caused by both pressure and volume overload. Pressure hypertrophy was induced by an aortic constriction; volume hypertrophy was induced by an iron-copper deficiency (anemia). The ventricular weight was increased by 34% in the pressure-hypertrophied hearts at the end of 6 weeks. The ventricular weight was increased by 54% in the volume-hypertrophied hearts at the end of 3 months. A potassium arrest-formalin fixation technique was used to produce a "diastole-like" ventricle. In the pressure-hypertrophied ventricle, the ventricular wall thickness and external radii were significantly increased, whereas the valve-to-apex distance and internal radii remained unchanged. We also found that in the volume-hypertrophied ventricle there was an increase in the valve-to-apex distance, external radii, internal radii, and wall thickness. Although external and internal dimensions increased, the ventricular shape did not change significantly in the volume-hypertrophied ventricle.


Mechanisms of cellular enzyme release. II. Inhibition of sarcolemmal enzymes by myopathy-inducing agents. A defective membrane mechanism has been suggested [Arch. Neurol. 33, 315 (1976)] for the pathogenesis of Duchenne muscular dystrophy. The characteristic clinical and biological findings, including leakage of cellular enzymes into the serum in the disease, have been duplicated by the imipramine/serotonin rat myopathy model. Sarcolemma was prepared from quadriceps femoris muscles of control and myopathy-affected animals. The activities of sarcolemmal adenosinetriphosphatase and acetylcholinesterase were inhibited in vitro by imipramine and serotonin. The inhibition by imipramine of these sarcolemma-bound enzyme systems decreased the Vmax and increased the Km. This mixed type of inhibition is consistent with an imipramine-induced interference at these enzyme sites and a disruption of lipid-protein interrelations. We hypothesize that such conformational membrane changes might contribute to the leakage of macromolecules such as enzymes from the cell interior.


Acoustic impedance measurement in the otological assessment of multiply handicapped children. This survey investigated the use of acoustic impedance measurement on a population of multiply handicapped children and correlated the results with otoscopy. Thirty-one children were tested with an impedance meter studying tympanometry, tympanic membrane compliance, and acoustic reflex. Otoscopic examination was also scheduled for each child. The results show that 42% of children tested had some kind of middle-ear problem, mostly unrecognized at the time of this survey. There was 100% agreement between impedance and otoscopic findings. Apparently impedance audiometry can be useful for identifying middle-ear disorders in children who cannot be tested by traditional audiometric methods.


Koro in a Yemenite and a Georgian Jewish Immigrant. Koro is a syndrome consisting of fears that the male organ is withdrawing into the abdominal cavity and ultimately this will lead to death. The syndrome is observed among inhabitants of South East Asia and seems to be extremely rare outside that geographical area. In this paper one case of koro is described in a Yemenite Jewish immigrant and another in a Georgian Jewish immigrant to Israel. Some of the theoretical aspects and implications of the koro syndrome are discussed.


A comparison of bovine nasal cartilage proteoglycan core protein produced by chondroitinase and hyaluronidase: the possible role of protease contaminants. The basic subunit of cartilage proteoglycan consists of multiple glycosaminoglycan chains covalently attached to a core protein. It is unclear as to whether there is a single core protein or multiple different core proteins, since previous studies using either chondroitinase or testicular hyaluronidase to enzymatically remove chondroitin sulfate side chains from the proteoglycan subunit have yielded conflicting results. In the present study, a chondroitinase-produced core protein preparation isolated as a single peak on Sepharose gel chromatography was found to contain at least two immunologically distinct components. Hyaluronidase-produced core protein from the same proteoglycan subunit fraction was found to contain multiple components nearly all of which were smaller than the components in the chondroitinase digest. A possible explanation of these findings is that they resulted from proteolytic degradation of the core protein in the course of the enzymatic removal of its chondroitin sulfate. The presence of small amounts of protease contaminants in several commercial chondroitinase and hyaluronidase preparations was detected by an extremely sensitive radioassay. Until proteases can be rigorously excluded from enzyme preparations used to degrade the proteoglycan subunit, it will not be possible to determine whether it consists of a single or several different core proteins.


Isolation and purification of a small molecular weight hydroxyproline-containing structural glycopeptide from early mammalian granulation tissue. A small molecular weight structural glycopeptide was solubilized after collagenase digestion of the connective tissue capsule surrounding the 5-day sponge-implant of the rat. The major amino acids are one residue each of aspartic and glutamic acids, proline, hydroxyproline and alanine and two residues of glycine, and the carbohydrates are one residue each of glucose, xylose and hexosamine and two residues of mannose. The sum of the amino acid and carbohydrate residues gives a molecular weight of 1635. Dansylation of the glycopeptide produces a single strongly fluorescent yellow-orange amino-terminal spot, not positively identified. The solubilization of the granuloma glycopeptide by collagenase and its composition are suggestive of its association with an immature form of collagen in early granulation tissue.


Effect of low dosage schedule of D-penicillamine on collagen cross-linking in a nine week immobilized rabbit knee. Rabbits immobilized with one leg pinned in a fully flexed position, were treated with 60 mg of D-penicillamine/kg three times a week for 9 weeks. After this period of immobilization the animals were sacrificed and the periarticular connective tissue collagen reduced with [3H] NaBH4 for determination of collagen cross-links. There was a significant decrease in the formation of cross-links in the control and immobilized knee in the treated animals as compared to the untreated animals. Penicillamine also blocked the increase in DHLNL, HLNL and HHMD previously demonstrated in immobilized rabbit knees. This is the first demonstration that in vivo administration of penicillamine directly inhibits cross-linking reactions in collagen.


The effects of ethanol on myocardial mitochondrial and sarcoplasmic ATPase in rats. The effects of the ethanol on the activities of mitochondrial and sarcoplasmic Mg2+-Ca2+-activated ATPase in rat myocardium were studied on two experimental alcoholic models: a) in an acute model - after a single dose of ethanol, 250 mg/100g of body weight; b) in a chronic model - after daily administration of the same dose for 10 weeks. In the acute model the ATPase activities sank both in mitochondria and sarcoplasmic reticulum. In the chronic model an opposite tendency was observed: in both subcellular organelles the ATPase activities moderately rose. The findings are assessed from the aspect of the dynamics of myocardial metabolic changes in dependence on the duration of action of ethanol upon the myocardium. It is stressed that acute symptoms of the ethanol action on myocardial metabolic processes are no adequate basis for drawing conclusions about the mechanism of myocardial lesion and the development of ethanol-induced mycardiopathy in chronic alcoholics.


Protein changes in frozen fish.

Protein changes in frozen fish. Storage of frozen fish brings about a decrease of extractability of myofibrillar proteins. There is also deterioration of the texture and functional properties of the flesh. In model systems, aggregation of myosin, actin, tropomyosin, and whole myofibrils have been described. These changes are caused by concurrent action of partial dehydration due to the freezing out of water, exposure of the proteins to inorganic salts which are concentrated in the remaining nonfrozen fluid, interactions with free fatty acids liberated from phospholipids and with lipid oxidation products, and cross-linking by formaldehyde produced in some species of fish as a result of enzymic decomposition of trimethylamine oxide. The extent of protein alterations increases with time and temperature of storage as well as with advanced disintegration of the tissues and intermixing of their components. The role played by the individual factors and the significance of different types of bonds, i.e., hydrophobic adherences, ionic bonds, and covalent cross-links in particular cases are not yet fully disclosed. Retardation of the deteriorative changes of proteins in frozen fish is possible by avoiding high storage temperatures and oxidation of lipids, removing hematin compounds and other constituents promoting cross-linking reactions, and by adding cryoprotectors like sugars, several organic acids, amino acids, or peptides.


Clinically silent pericardial effusions in patients on long-term hemodialysis. Pericardial effusions in hemodialysis. The danger of cardiac tamponade occurring when pericarditis is accompanied by pericardial effusion, as opposed merely to the presence of a friction rub without effusion, has been unclear. Forty patients on hemodialysis were studied by physical examination, chest x-ray film, and echocardiography for evidence of pericarditis and pericardial effusion. Only two patients developed a friction rub during the study and were placed on regional heparin. Ten of 11 patients who were positive on echocardiogram for pericardial effusion had unremarkable physical examinations. These 11 patients had cardiomegaly as noted on chest x-ray examination. Eighteen of 25 patients without effusion also had cardiomegaly on chest x-ray film. No patient remaining on systemic heparin and having a pericardial effusion developed cardiovascular complications during hemodialysis. This study suggests that while many patients on longterm hemodialysis have pericardial effusion undiagnosed on the basis of physical examination, but noted on echocardiogram, special precautions to prevent tamponade during hemodialysis are not necessary. Also, posterior-anterior chest x-ray film showing a normal-sized heart will usually exclude significant pericardial effusion.


[Experience report on the intraoperative use of the ultrasonic Doppler Probe]. Experiences in the intraoperative use of an ultrasonic doppler-flowmeter are described. The method can be helpful when performing various operations within the field of generalsurgery and vascular surgery.


Histochemistry of inferior oblique muscle in a case of congenital third nerve palsy. Material obtained from the inferior oblique muscle proved to give the best preparations for histological and histochemical investigation. The normal aspects of inferior oblique muscle are discussed. The usefulness of inferior oblique muscle biopsy in pathological material is illustrated by a biopsy of a 6-year-old boy operated on for strabismus due to congenital third nerve palsy.


[Effect of talliblastine on several hematologic indices]. The authors carried out a study on the blood formation of healthy experimental animals after treatment with the potential antitumours agent taliblastine in view of clarifying its influence. The experiments were undertaken on inbred mice DVA/2 and rats on the Wistar strain. Taliblastine was administered intraperitonealy: in ice-aingly and in a 5-day course, but in rats only in a 5-day course. The substance was administered after single treatment with doses of LD50 maximal threshold limit dose (MTD), but in the dourses with MTD and 1/2 of MTD. They examined the changes in hemoglobin, number of leucocytes, erythrocytes, thrombocytes and reticulocytes, but in rats-the number of bone-marrow cells as well. The values of hematological indices were determined 24 hours, 10 and 30 days after conclusion of the administration of taliblastine. It was established that taliblastine did not affect substantialy the values of hemoglobine and caused transitory changes in the absolute number of leucocytes and lymphocytes.


Adenosine triphosphatase activity associated with purified cholinergic synaptic vesicles of Torpedo marmorata. A rapid method for purifying Torpedo electric organ vesicles is described, which employs an isoosmotic continuous sucrose-glycine gradient followed by chromagography on CPG-10-3000 porous glass beads. The synaptic vesicles have a buoyant density of 1.057 g/ml. The purified vesicles are free of cholinesterase, lactate dehydrogenase and Na+, K+-stimulated ATPase activity. They contain a  ouabaininsensitive, Na+, K+-inhibited, Mg2+, Ca2+-stimulated ATPase activity. This is further stimulated by acetylcholine but not by choline.


The isolation and characterization of differentially phosphorylated fractions of phosphofructokinase from rabbit skeletal muscle. A preparation of phosphofructokinase from rabbit skeletal muscle is described which exploits the association-dissociation properties of the enzyme. Phosphofructokinase to prepared is partially phosphorylated and may be fractioned into three distinct species with sedimentation coefficients of 30 S, 18 S and 13 S by chromatography of agarose gels, hydroxyapatite or DEAE-cellulose. Measurements of alkali-labile phosphate content (phosphoserine and/or phosphothreonine) show that fractions consisting almost exclusively of 30-S species and fractions consisting predominantly of 18-S and 13-S species contain approximately 0.15 and 0.29 mol of phosphate per phosphofructokinase monomer (Mr = 80000) respectively. The results are interpreted in terms of at least two 13-S components which differ in their phosphate contents and also in their self-association properties. The possible significance of phosphorylation is discussed.


T cells which proliferate in response to concanavalin A include cells which proliferate in mixed leucocyte reactions. Selection in long-term culture of alloreactive T cells, by successive in vitro restimulation with semi-allogeneic cells, results in primed responder cell populations which maintain full proliferative reactivity to allogeneic cells as well as to the T cell mitogens concanavalin A (Con A) and phytohemagglutinin (PHA) but are depleted of cells which can effect target cell destruction in either a specific or nonspecific manner. Con A-induced T cell blasts (selected by velocity sedimentation) can revert to small resting lymphocytes in the presence of inert "filler" cells. Con A blasts which have reverted, readily proliferate in response to Con A or allogeneic stimulator cells but are largely depleted of effector killer cells and PHA-responsive cells.


Lack of effect on brain stem and cerebral cortex Na+, K+-ATPase during heart block produced by chronic digoxin treatment. The autonomic nervous system has been shown to play an important role in digitalis toxicity. In order to determine whether the central nervous system could be digitalis' site of action, the effect of chronic treatment with toxic doses of digitalis on brain Na+,K+-ATPase was studied in the dog. After four weeks of digoxin treatment, Na+,K+-ATPase activity of the brainstem or cerebral cortex was unaffected at the time when digitalis toxicity (heart block) was apparent. ATP-dependent (3H)-ouabain binding to these tissues was also unaffected indicating that a significant occupancy of brain Na+,K+-ATPase by digoxin did not occur during chronic drug treatment. In contrast, cardiac Na+,K+-ATPase was markedly inhibited with the concomitant binding of digoxin to the enzyme. Since Na+,K+-ATPase is the most digitalis sensitive system identified to date, it appears that digoxin does not affect neuronal function directly.


Echocardiographic diagnosis of a pulmonary artery aneurysm. Aneurysm of the pulmonary artery is an uncommon anomaly which may be very difficult to diagnose without angiography. The present report demonstrates that the echocardiographic diagnosis of a pulmonary artery aneurysm is feasible. Correlation between chest X-ray, echocardiogram, cardiac catheterization, angiography and necropsy is presented. Several echocardiographic features of the pulmonic valve motion in pulmonary hypertension, not previously described, are discussed.


Comparison of polyglactic and polyglycolic acid sutures in reproductive tissue. Polyglactic acid and polyglycolic acid suture materials were compared in rat uterine and abdominal wall tissues for inflammatory response and tissue fibrosis. By 90 days after surgery, the tissue inflammatory reaction and fibrosis were significantly less in response to polyglactic acid suture (Vicryl) in both uterus and skin as compared with the response to polyglycolic acid (Dexon). In addition, the over-all tissue response in skin was significantly greater than that in uterus for both suture materials. The potential importance of tissue fibrosis--particularly in oviductal surgery, over and above the formation of adhesions between one organ and another--is emphasized. It is concluded that (1) the magnitude of tissue response to suture material varies for different tissues, (2) the degree of tissue wall fibrosis does not necessarily correspond to external tissue adhesions, (3) adhesions are maximal at the surgical knots regardless of the suture material used, and (4) polyglactic acid suture material may be preferable to polyglycolic acid suture material for infertility surgery, in which a minimum of tissue reaction is imperative.


[Mechanism of the hypobilirubinemic action of nicofezon]. The effect of nicophesone on the processes of cojugation and alkaline phosphatase was studied in tests staged on albino rats and rabbits poisoned with carbon tetrachloride (CCl4). Nicophesone is shown to stimulate conjugation of bilirubin with glucoronic and sulphuric acids and to depress the activity of alkaline phosphatase. The data obtained are considered inasmuch as they concern the hypobilirubinemic action of nicophesone.


[Ultrastructural localization of adenosine triphosphatase activity in the proximal kidney tubules of white rats]. In white rats, the ferment topography of Mg+2 and (Na+ + K+)-activated ATPh-ses in proximal canaliculi was studied with the aid of the ultrastructural cytochemistry. The final product of the fermentative reaction (PhHPO4) in the form of small dense granuli is positioned on the duplicate folds of epithelial cells, the cells limiting the brush border micropiles, and on invaginations of the apical plasmalemme at the micropiles base. For (Na+ %K+)-activated ATPh-ses a localisation of the reaction product was determined in the canaliculi vessels.


Cellular cooperation at the level of antibody-producing cells in chickens. The effect of in vitro cocultivation of immune and non-immune syngeneic lymphoid cells on the antibody response was studied in chickens. Cocultivation of immune and non-immune spleen cells did not affect significantly the PFC numbers. Substantial increase in PFC was observed in mixed cultures of immune spleen cells with non-immune bone marrow cells. The existence of an enhancing effect of non-immune cells on the antibody-producing cells, which had been earlier described in mice, was observed in a phylogenetically distant species. Therefore it seems probable that it exists in most, if not all, homoiotherm animal species.


Psuedolymphoma syndrome.

Psuedolymphoma syndrome. A transient dermatologic abnormality which gave a histologic picture compatible with malignant lymphoma is described. The clinical manifestations included diarrhea, hepatosplenomegaly, moderate lymph node enlargement without histopathological changes and evidence of involvement of the reticulo-endothelial system. Both the dermatologic and gastrointestinal disorders appeared approximately 1 year after discontinuation of treatment with diphenylhydantoin, which the patient had taken for 2 years. The possible association between the transient abnormality and the previous diphenylhydantoin therapy justifies the classification of this case as 'pseudolymphoma syndrome'.


[Statistical evaluation of vectorcardiographic criteria for the diagnosis of posterior myocardial necrosis (author's transl)]. 85 vectorcardiograms, characterized by anteriorly oriented horizontal ventricular loop, were analized in order to evaluate if this pattern can be considered diagnostic at all for strictly posterior myocardial infarction. Records were divided into two groups according to positivity or negativity of clinical history for myocardial infarction. Records consistent with right ventricular hypertrophy were ruled out. Nine parameters related to ventricular depolarization and repolarization were measured and the values were submitted to statistical analysis. The study showed that:--selection of the two groups was correct, because of first group's homogeneity and control group's unhomogeneity;--mean values of the nine parameters did not differ significantly in the two groups;--single parameter into each group was unrelated to others. As a consequence, criteria at the time in use for diagnosis of strictly posterior myocardial infarction seem not to be useful, as other conditions beside myocardial infarction or right ventricular hypertrophy are able to determine the same vectorcardiographic pattern.


[A new surgical technique for treatments of left ventricular outflow tract obstructions. Report of a case and reexamination of our experience (author's transl)]. Aortic valvotomy, aortic valve replacement, resection of the subaortic membrane and aortic patch angioplasty do not always provide satisfactory relief of left ventricular outflow tract obstruction. From August 1975 through January 1977, 15 patients underwent creation of a left ventricular vent by means of a Dacron fabric conduit containing a heterograft valve going from the left ventricle to the supraceliac portion of the abdominal aorta (Cooley's point). All patients had valvular, subvalvular, supravalvular aortic stenosis, or combined defects. One patient had severe hemolytic anemia following aortic valve replacement. Eleven of our 15 patients had undergone one or more previous surgical procedures for relief of aortic stenosis. The operative mortality has been one patient. At postoperative catheterization, there was a normalization of pressure gradients, and the blood flow through the conduit was 40% of the cardiac output. An interesting case of supravalvular aortic stenosis associated with stenosis of the vessels of the arch is presented.


[Sterilization by partial resection of the oviducts via bipolar electrocoagulation (author's transl)]. A total of 2,357 sterilizations via bipolar coagulation was effected at the University Clinics of Gynecology at Basel and Tübingen since 1972 and 1975 respectively. Of these, 2,143 were effected via laparoscopy. The following complications occurred which required laparotomy: in two cases, hemorrhages from the abdominal cavity as a result of the second puncture, in one case from the mesosalpinx, and, as a late complication, two extrauterine pregnancies. The method failed in the case of the two extrauterine pregnancies and possibly also in the case of two doubtful intrauterine pregnancies (0.8--1.7 per thousand). Today this surgical measure is being performed in Tübingen exclusively by the single-puncture technique an increasingly under local anesthesia. The advantages of the method are an accurately determinable coagulation zone, sparing of the mesosalpinx, low incidence of complications, absence of life-threatening incidents, and low failure quota.


Mucosubstances in the human skeletal muscles. With histomchemical, and electronmicroscopic-histochemical methods two types of human skeletal muscle fibres were established. The first type of muscle fibres does not contain acidic mucosubstances. The staining reactions and cellulase digestion indicate that, the neutral polysaccharides are cellulose-like substances. The second type of fibres contains only acidic mucosubstances, hyaluronic acid and chondroitine sulphate. The author suggests that the mucosubstances have joint function. These polysaccharides contributed to the jointing both of myofibrils and sarcomers. The polysaccharides can be exhibited in the joint points of contractile elements. In mechanical injury this point became disintegrated.


Histochemical studies on the mucopolysaccharides of the developing chick kidney. II. Mucopolysaccharides of the collecting tubules and excretory ducts. Mucopolysaccharides of the collecting tubules and excretory ducts of developing chick kidney were studied histochemically to elucidate the relations, between the changing chemical properties of these compounds and excretory processes during pronephric, mesonephric and metanephric developmental stages. At the pronephric ammonotelic stage the collecting tubules contain only neutral mucopolysaccharides, whereas at the mesonephric ureotelic stage neutral mucopoly saccharides are found till the 9th day. Sialic acids make their appearance in luminal border regions of the mesonephric collecting tubules from the 9th day on, their concentration being highest on the 15th day. Hyaluronic acid is observed from the 16th day on. Its concentration is predominant in hatched young birds and increases with age. The physiological significance of alterations in the mucopolysaccharides contents is discussed.


Histochemistry of the utero-vaginal junction with special reference to the sperm-host glands in the oviduct of the domestic duck. The "utero-vaginal junction" that connects the uterus and vagina of duck oviduct differs histologically and histochemically from the adjacent zones. The region is characterised by low, somewhat longitudinally arranged mucosal folds, lined by tall columner cilliated cells with apical nuclei alternating with mucous secreting goblet cells containing basal nuclei. Within the propria of the mucosa there are numerous tubular glands--the sperm host glands--which are responsible for sperm storage after copulation. The goblet cells of the utero-vaginal junction contain an admixture of neutral, sulfated and nonsulfated acid mucopolysaccharides, while the sperm host glands are devoid of any mucopolysaccharide and secrete complex lipoidal materials and exhibit intense acid phosphatase activity. The functional significance of the secretory materials of the sperm host glands have been discussed in the light of sperm release mechanism.


Histochemistry of mucosubstances in the mantle of fresh water mussel, Parreysia corrugata var. nagpoorensis (Lea) II. Mucosubstances of the middle marginal fold. The results of the various histochemical reactions on mucosubstances indicate that in the middle fold of the mantle edge two types of mucus cells exist, one producing sulphomucins and the other neutral mucosubstances. The cells secreting neutral mucosubstances are few in number. The sulphated mucus is strongly alcianophilic. The alcianophilia persists when the tissues are stained with alcian blue in concentration up to 0-5 M magnesium chloride. Testicular hyaluronidase has no effect on the staining pattern of the mucus.


[Mammography and age (author's transl)]. Following Egan (2), Gros et al. (8,9), Ingleby and Gershon-Cohen (11) and Witten (18) the appearances of mammograms were divided into four groups: I. Dense, II. Atrophic, III. Shrunken-fibrous and IV. Cotton-wool, diffuse. In young women up to the age of 30 a dense structure is the most common (74.8%). On the basis of available statistics, the use of mammography for routine detection of carcinoma of the breast cannot be justified in women under the age of 30; not only is this condition very rare at this age, but proper evaluation of the relevant signs is only possible in one woman in four because of the structure of the breast. On the other hand, if there is any suspicion of the presence of a carcinoma, then mammography should be carried out and additional forms of examination, such as thermography and xerography, should not be neglected.


[Clinical-radiological evaluation of barium sulfate suspension in double contrast examination (author's transl)]. In four selected, commercial barium sulfate preparations, viscosity, surface tension and particle size were measured; examined were agglomeration, sedimentation and homogeneity in vitro. For the clinical-radiological examination, an evaluation scheme has been developed containing the following criteria: uniformity and thickness of contrast medium layer, demonstration of the areae gastricae, clearness of gastric contour, and vesiculation. A blind study, encompassing about 100 double contrast examinations of each preparation, ascertained the usability of this scheme. Significant differences could be detected among the four contrast media. The barium sulfate suspension which was rated the highest lay within a viscosity range of 50-90 cP, had a particle diameter of 0.2-1.6 micrometer and showed the least degree of agglomeration. Results of the in vitro examinations, when compared to the clinical tests, lead to the conclusion that both, gastric mucous and contrast medium additives, may exert considerable influence on the pictorial quality of the above defined criteria.


[Radiological bone patterns compared with pathological and anatomical appearances (author's transl)]. In the present study, radiographs of the skeleton were compared with relevant pathological sections; in this way some information could be more clearly brought out and changes in the structure and contours of bones could be classified. The radiographs were compared with photographs of the actual specimen. For this purpose either plain radiographs or tomographic cuts were used. In addition, bone sections were studied by a radiographic magnification method in order to improve detail recognition.


[Pathological bone structure (author's transl)]. The pathological and anatomical basis of various radiographic changes in a number of diseases of the skeleton is discussed. The conditions include abnormalities of bone formation and bone removal, including osteoporosis, fibrous dysplasia, osteomyelosclerosis, osteitis deformans, posttraumatic changes and ossifying periostitis. The form and structure of the pathological and anatomical substrate determine the radiological appearances and are essential for arriving at a diagnosis.


[Köhler's "tear drop". Relationship between the radiologic picture and anatomic substrate]. The diagnostic value and anatomical basis of Köhler's "tear drop" has been in doubt. By means of radiographs of the right side of the pelvis taken with various degrees of rotation tilt, and by X-raying serial sections, it was possible to demonstrate how this appearance is produced. It was shown which parts of the pelvis contribute to the "tear drop". This simplifies an understanding of the radiographic appearances of the pelvis in the region of the hip joint.


[The radiographic appearances of the acromio-clavicular joints in old age (author's transl)]. The author studied 200 shoulders in 100 normal persons over the age of 60 and found the following changes in the acromioclavicular joint due to old age: simple sclerosis of the joint surfaces, some new bone formation, hypertrophy of the acromial end of the clavicle, uneveness of the acromion and caudal dislocation of the clavicle. Marginal changes of the clavicle at the point of attachment of the coraco-clavicular ligament are also due to old age and are not a sign of rheumatoid arthritis.


[Soft tissue changes in pseudo-pseudo-hypoparathyroidism (author's transl)]. The authors have demonstrated subcutaneous calcification which is characteristic for pseudo-pseudo-hypoparathyroidism (pph) and which is best shown by xero-radiography. If the presence of pph is suspected, or if it needs to be distinguished from some other condition, it is valuable to obtain xerograms of the soft tissues of the extremities. In pph the subcutaneous calcification is in all probability derived from the skeleton.


Achondrogenesis, type I.

Achondrogenesis, type I. Achondrogenesis is a rare form of lethal dwarfism. Usually discovered post partum, in a few instances only it has been diagnosed in utero. One such a case is here presented together with differential diagnosis with other forms of lethal dwarfism.


[The arteriographic appearances of the hypothenar hammer syndrome (author's transl)]. The characteristic angiographic appearances found in the superficial palmar arch are described in five cases of the hypothenar hammer syndrome. The symptoms depend on the anatomical peculiarities of the arteries in the hand, the extent of the causative lesion and the condition of the peripheral digital arteries. The condition is probably more common than is thought. It can be differentiated from other vascular abnormalities of the hand by angiography.


[The primitive otic artery, a very rare, persistent, primitive artery (author's transl)]. A very rare carotid-basilar anastomosis, the primitive otic artery, is described. In the presence of hypoplasia of the vertebral and basilar arteries and dorsal interruption of the circle of Willis, an attempt is made to explain the situation embryologically and to discuss the disadvantages associated with this condition.


[Demonstration of the skull base by computed tomography (author's transl)]. In addition to showing the structures of the brain, it is possible with computed tomography to demonstrate the bone of the skull. For this the window width has to be increased, or one has to use additive photographic manipulation (integral photography). The clinical value of views of the skull base is illustrated. The advantages of computed tomography compared with conventional radiographic technique are stressed.


[First clinical experience with axial emission computer tomography (author's transl)]. 104 examinations were carried out on 81 patients with a new commercial emission computer tomographic scanner. The technique provides additional information in half the brain scans, but only in one third of liver scans. No indications for this examination could be discovered in a relatively small clinical material for the examination of the kidneys or the skeleton. Other organs were not included in this assessment. The authors confirm reports in the literature which show that this method is not particularly valuable for finding pathology. It finds its application in conforming or excluding suspected abnormalities and for better localisation and delineation of lesions, thereby aiding differential diagnosis. However, further technical improvements and increased clinical experiency may increase the applicability of the technique.


[Improved scintigraphic imaging of the adrenals with 6beta-131I-iodomethyl-cholesterol (author's transl)]. Scintigraphic visualization of the adrenals is greatly improved after substitution of 131I-19-I-Cholesterol by 6beta-131I-Iodomethyl-Cholesterol which was first seen as a radiochemical impurity during the synthesis of 131I-19-Iodocholesterol. The interpretation of the scintigraphic imaging is further improved by application of a cross section through the visualized adrenals. In an animal experiment the radioactivity of the adrenals and to a less degree that of different tissues was found up to four times higher after injection of 6beta-131I-Iodomethyl-Cholesterol than with the applications of 131I-19-Iodocholesterol. The high affinity of the tissue for 6beta-131I-Iodomethyl-Cholesterol results in high radiation exposure of the patients. However, since less activity of 6beta-131I-Iodomethyl-Cholesterol is necessary for visualization of the adrenals the overall radiation is also less than with the application of 131I-Iodocholesterol. Higher stability of the labelled molecule is suggested to be responsible for improved scintigraphic imaging of the adrenals.


[Sequential scintigraphy in the investigation of changes in renal function during pregnancy (author's transl)]. Changes in renal function during pregnancy were studied by sequential scintigraphy in 16 women with normal kidneys. The examinations were carried out during the first and third trimesters. Time activity curves were obtained by a computer-assisted storage and analysis procedure, using a "region of interest" technique for renal parenchyma and renal pelvis. The study included topographic changes and the evaluation of 13 semi-quantitative parameters related to the renal parenchyma and of nine parameters related to the renal pelvis. The results were demonstrated graphically. The predominantly rightsided changes in renal function during the third trimester are discussed in relation to the relevant literature.


[Inhibition malformation of the female genitaltract combined with gynatresia on one side (author's transl)]. Reduplication malformations of uterus and vagina are caused either by absence or inkomplete fusion of the paired Müller's ducts. When this occurs in conjunction with gynatresia it's often not diagnosed until many years after the menarche. Even by hysterosalpingography only few cases of combined inhibition malformation of the female genitaltract are diagnosed. The radiological findings in a patient with uterus bicornis bicollis and vagina septa unilateralis associated with unilateral renal aplasia are described.


[Computerised tomography of the trunk-a first report (author's transl)]. One thousand patients have been examined with the body scanner and a preliminary report is presented on the value of this method in the examination of abdomen and the thorax. Unlike other radiological procedures, computerised tomography is often able to demonstrate disease states directly and not only as a result of their effect on neighbouring structures. It provides a non-invasive means for achieving a diagnosis, for therapy planning and for control of treatment of space-occupying lesions within the trunk. Its role can only be defined after a systematic comparision with conventional diagnostic procedures.


[Whole body computer tomography in the planning and follow-up of radiation therapy (author's transl)]. Whole body computer tomography provides accurate transverse sections of the areas to be irradiated in the position used for irradiation. The method has been used in 167 patients with malignant tumours in various parts of the body on a routine basis. Actual-size cuts allowed accurate delineation of the tumour or of the area to be irradiated with its neighbouring organs in 135 patients; in 66 patients CT scanning probided additional information regarding the extent of the tumour. Follow-up examinations carried out in 29 patients proved effective in showing the results of treatment. The introduction of the whole body scanner into radiation therapy has improbed radiation planning when compared with conventional methods because of the more accurate delineation of the area to be irradiated and of the surrounding organs.


[Polycystic degeneration of parenchymatous organs in the abdomen or malignant tumour? Computer tomographic investigations]. The report deals with the computer tomographic investigation of a space-occupying intra-abdominal process in which there were polycystic lesions of the liver, kidneys and pancreas. CT scanning combined with biochemical investigations provided clear proof of the presence of cysts, their morphology and extent. The possibility of the neoplasm could be excluded; it proved unnecessary to carry out any invasive procedures and a laparotomy also became unneccessary.


[Cholangiographic appearance of the intraphepatic bile ducts (author's transl)]. Percutaneous transphepatic cholangiography patterns of intrahepatic bile ducts in 134 patients were analysed. The majority of case material consisted of gallstone disease and complications of its surgery (44 cases) and secondary or primary neoplasms of the liver (40 cases). A satisfactory filling of the peripheral bile ducts was obtained after drainage of bile and with an injection of somewhat increased quantities of contrast medium into the biliary system. PTC was found to be highly effective in detecting space occupying lesions of neoplastic origin i.e. sensitivity-90 percent, specificity-95 percent, predictive value of a positive result-88 percent. PTC patterns of liver cirrhosis with concommitant extraheptic obstruction, chronic cholangitis, congenital ectasia of the bile ducts, liver abscesses are described. Although PTC is aimed primarily at the detection of nature and level of the extraheptic biliary obstruction there is no essential reason for neglecting its possibilities in recognizing intrahepatic disease.


[Hepatobiliary scintigraphy with (99m)Tc-diethyl-IDA (author's transl)]. A short description of the pharmacology and kinetics of (99m)Tc-diethyl-IDA is given; our experience with this new radiopharmaceutical as a functional test of liver and bile ducts is illustrated by a number of typical examples. The ability to use increased activity together with high resolution "low energy" detectors provides excellent detail recognition in the hepato-biliary system. In addition, sequential scintigraphy with its resulting time-activity curves provides information regarding the function of the hepatocytes. The significance of hepatobiliary scintigraphy is its ability to distinguish between parenchymal and obstructive jaundice.


[Hepato-biliary sequential scintigraphy-developments, methods and recommendations for its clinical use. Experience in 350 patients with (99m)Tc-Solcoscint-Hepatobida (author's transl)]. Hepato-biliary sequential scintigraphy is an isotope method in the diagnosis of liver and biliary disease. The value and accuracy of the diagnostic information depends on the development of high resolution gamma cameras and new radiopharmaceuticals; these are described briefly. Our investigations on 350 patients were carried out (99m)Tc-Soloscint-Hepatobida. A number of clinical examples illustrate the scope and value of the examination; from these the indications can be deduced. Hepto-biliary scintigraphy will not replace proven gastro-enterological and radiological methods of investigation, but may be added to these in order to obtain rapid and reliable information. This will be of assitance both to the physician and the patient in deciding which of the more complex methods should be employed next.


[Sonography and conventional radiography for post-operative local infections in the upper abdomen (author's transl)]. The findings on sonography and radiography were compared in 32 patients with abscesses or empyemas in the upper abdomen following surgery. Sonography demonstrated each of eight liver abscesses, eleven of 21 upper abdominal empyemas and three abscesses or empyemas in other positions. Corresponding figures for radiography were five out of eight liver abscesses, 16 out of 21 upper abdominal empyemas and one of the remaining lesions. The methods are complimentary. Sonography demonstrates fluidfilled cavities, radiography shows particularly the presence of extraluminal gas. Recommendations are made for the proper sequence of these diagnostic measures.


[Fine needle puncture for antegrade pyelography and percutaneous nephropyelostomy (author's transl)]. A method for antegrade pyelography and pyelonephrostomy is described using a long, thin needle with a shorter, wider Seldinger cannula. Puncture with a thin needle reduces the risk during antegrade demonstration of the upper urinary tract and can also be used for puncturing collecting systems which are only slightly dilated. When necessary, the Seldinger cannula can be advanced over the needle; subsequently, with the aid of a J-wire, a nephropyelostomy can be performed. The method has been used in 20 patients. In one case of intended bilateral nephropyelostomy the puncture was successful on only one side. There were no serious complications. In only one patient a small perirenal haematoma was found at operation.


[Radiological findings in severe renal trauma during childhood (author's transl)]. The radiological findings in 21 children aged between four and 14 years, suffering from severe renal trauma, are described. Various radiological signs are described and their value and frequency are discussed. Plain films provide only non-specific evidence. Contrast extravasation during an excretions urogram is the only reliable sign of rupture. Immediate angiography in order to exclude vascular damage is indicated if there is no contrast excretion by the damaged kidney. Angiography at a later date should be carried out if there is persistent haematuria, in  cases of doubt regarding the excretion urogram, or when it is impossible to decide on the basis of the excretion urogram whether operative or conservative treatment is indicated. Regular observation is necessary, both after operative and conservative treatment, in order to recognise complications at an early stage.


[Erogotism as a cuase of abnormal peripheral arterial flow (author's transl)]. Ergotism is seen rarely as a complication of the treatment of migraine with drugs containing egotamine. The prognosis is good if the disease is recognised at an early stage and ergotamine medication is discontinued. A case of ergotism studied angiographically is described. The patient was a 46-year old woman who developed severe vascular spasm in pelvic and lower limb arteries within four weeks of taking 100 mg. erotamine tartrate. The changes disappeared completely once the ergotamine preparation was stopped.


[Gorlin-Cohen syndrome (frontometaphyseal dysplasia)]. The classical case of a Gorlin-Cohen-Syndrom (= fronto-metaphyseal dysplasis=FMD) is presented. The disease has been mentioned in literature for the first time in 1969 and has been described four times up to now. The characteristic features of this syndrome are very prominent supraorbital ridges with generalized bone dysplasia and joint deformities.


[Arthrography of the hand in an accident clinic (author's transl)]. By 115 arthrographies of the wrist in patients with posttraumatic pain, the importance of the method for diagnosis in injured patients is discussed. It was possible to detect 29 cases with a lesion of the articular disc, 4 patients with ruptured radiocarpal joint and 3 cases with an injury of the distal radioulnar joint. The technique of filling the wrist is described and different manifestations of "normal" arthrogramms are exhibited.


[Arthrography and ultrasound scanning for the detection of Baker cysts (author's transl)]. 23 knee joints with clinical evidence of Baker's cysts have been studied in comparison by arthrography and ultrasound-scanning. In 20 knees, there was correlation between arthrographic and sonographic findings. In 3 cases, a positive sonogram was not confirmed by arthrography. Possible causes for faulty interpretation and differential diagnoses have been discussed. We prefer ultrasound scanning for the diagnosis in inflammatory and degenerative joint diseases because it is entirely harmless. In cases of traumatic Baker cysts the arthrogram may reveal an additional meniscal injury.


[A simple device for examination of the ankle joint (author's transl)]. Ligamentous injuries of the ankle joint can be demonstrated radiologically by special techniques using lateral and A.P. projections. A device is described for obtaining the requisite views. It enables accurate projections to be achieved and a standardised technique is assured by applying a pull of constant weight. The problem of muscle spasm, resulting as a reflex from pain, is discussed.


[Possibilities and limits of xeroradiography. A statement (author's transl)]. Xeroradiography has proved to be of particular use for the examination of soft tissues. Due to great latitude and edge enhancement effect alterations of the splanchnocranium, the breast and the juxtaosseous soft tissues are especially well recognizable. Also for examination of larynx and trachea as well as certain soft tissues in the region of the neck this method is advantageous and proves its facilities to demonstrate diseases of the lungs and hilar region. On the contrary, alterations of the vertebral column, the basis of the skull and the abdomen cannot be examined by means of xeroradiography because of not justified radiation dosage. Xeroradiography offers no advantage for arthrography, angiography and examination of bones and skeletal parts of joints.


[Positive ventriculography with metrizamide (Amipaque)(author's transl)]. Our experience with the water-soluble contrast medium metrizamide (Amipaque) for ventriculography is described. No significant side effects were observed in our 27 patients, of whom 12 were children. A mild increase in CSF cell count disappeared after a few days. In 24 of the 27 patients, very good or adequate, and in any case diagnostic, information was obtained regarding the third and fourth ventricles, aqueduct, as well as sometimes the basal cisterns and outline of the cerebellum.


[Side effect of metrizamide and meglumine iocarmate in lumbar myelography. A prospective, parallel, interview study (author's transl)]. Side effects of metrizamide (Amipaque) and meglumine iocarmate (Dimer-X) were compared in lumbar myelography of 161 patients by a prospective, parallel, interview study. Severe complications were not observed. The percentage pairs of smaller complications were mainly in favour of Dimer-X, but in 2 of the 54 pairs only the difference did raise to a significant level. In one pair there were less symptomless patients after Amipaque-myelography (p less than 0.05) and in another pair the men had more headache after Amipaque-myelography (p less than 0.01). It may be possible to show with larger comparative series, that Amipaque has a tendency to cause more central-subcortical or meningeal-side effects than Dimer-X.


Influence of somatostatin on peripheral leucocyte count and granulation tissue in man and rats. Subcutaneous injection of synthetic protamin-zink-somatostatin completely prevents endotoxin-induced leucocytosis in normal rats. Piromen-induced elevated stab neutrophil, neutrophil and monocyte counts remain within the normal range during somatostatin administration. There is an inhibiting effect of synthetic protamin-zink-somatostatin on the wet weights of granulation tissue of cotton pellet granulomata, too. Incorporation of 35S-sulfate in sulfated mucopolysaccharides of granulation tissue in cotton pellet granulomata is not inhibited. Intravenous administration of synthetic linear somatostatin decreases stab neutrophil and neutrophil blood count in patients with acute bacterial leucocytosis. After the termination of somatostatin infusion a rebound phenomenon occurs. In healthy subjects lymphocyte count increases during somatostatin infusion. This effect can not be demonstrated in patients with bacterial leucocytosis.


Pseudopyogenic granuloma: enzyme histochemical and ultrastructural study. In an attempt to elucidate the nature of the abnormal vessels in pseudopyogenic granuloma, we performed enzyme histochemical analysis of 10 lesions and correlated them with electron microscopic findings in four lesions. The dermal vessels of pseudopyogenic granuloma possess voluminous endothelial cells demonstrating marked activity of various hydrolytic and respiratory enzymes. The alkaline phosphatase activity, however, is characteristically faint or absent. Electron microscopy reveals that mitochondria and the other common cytoplasmic organelles are abundant in the hypertrophic endothelial cells, probably reflecting increased metabolic cellular activity, whereas specific endothelial granules (Weibel-Palade bodies) are generally sparse in these abnormal cells. Other notable features occurring in several endothelial cells are cytoplasmic vacuolation and the acquisition in the subnuclear zone of dense bodies associated with fine actin-like filaments, 55 to 75 A thick. The latter change is interpreted as evidence of endothelial contraction. Classification of the abnormal dermal vessels using available criteria has proved difficult, but it is suggested that they arise predominantly from the venous side of the microcirculation and are mainly postcapillary venules, although involvement of both larger and smaller caliber vessels also occurs.


Meiosis I non-disjunction as the main cause of trisomy 21. The relative roles of Meiosis I and Meiosis II non-disjunctions in the causation of trisomy 21 have been assessed by analysing the distribution of polymorphic phenotypes of the chromosomes 21 in a group of individuals with Down's syndrome. The data suggest that the majority of cases of trisomy 21 are due to meiosis I non-disjunctions.


Serotype B staphylococcal bacteriophage singly converting staphylokinase. A new staphylococcal phage, Pphi-2, which could convert the capability of producing staphylokinase, was isolated. The phage is different from the phage reported by Winkler and co-workers [Nature(London) 195:407-408, 1962; J. Gen. Microbiol. 39:321-333, 1965]. The former is a single-converting phage belonging to serotype B, but the latter is a serotype F phage capable of converting not only staphylokinase but also beta-hemolysin. Staphylokinase, i.e., the fibrinolysin produced by Staphylococcus aureus, therefore, can now be classified into three types. One is controlled by the genes on the bacterial chromosome, and the other two are mediated by lysogenic conversion by prophage Pphi-2 and the classical phage reported by Winkler and co-workers.


Use of transferrin-iron enterobactin complexes as the source of iron by serum-exposed bacteria. Two smooth and six rough strains of Salmonella typhimurium with progressively smaller amounts of sugar and protein in their outer membrane were tested for degree of virulence in normal and iron-injected mice and for ability to acquire iron in mammalian sera. The rate of mortality showed that bacterial virulence for mice was lowered with progressive decrease of outer-membrane sugar and protein. Iron injections increased the rate of mortality in mice infected either with smooth strains or with superficially rough strains but were without effect in mice infected with deep rough strains. In in vitro experiments, iron promoted with equal effectiveness the growth of all serum-exposed bacterial strains, whereas enterobactin (E) was much more effective in promoting the growth of smooth and superficial rough than in promoting that of deep rough strains. Various experiments showed that deep rough strains cannot grow in E-supplemented serum because they are not able to use the transferrin-iron-E complexes that E forms with transferrin-iron. This failure to use transferrin-iron-E complexes by deep rough strains was found to be due to the inability of these strains to absorb iron containing complexes to their outer membrane. Adsorption studies with chemically treated bacteria showed that the receptor of transferrin-iron-E or E-iron complexes is a protein of the outer membrane of bacterial cells.


Papular mucinosis.

Papular mucinosis. Most patients previously reported to have papular mucinosis had the generalized lichenoid papular form (scleromyxedema) with abnormal gamma globulin. Microscopic examination of affected skin showed increased acid mucopolysaccharides in the dermis and proliferation of fibroblasts. Our patient had the discrete form of papular mucinosis without abnormal serum gamma globulin but with increased amounts of acid mucopolysaccharides in the dermis and no proliferation of fibroblasts.


Primary and recurrent herpetic whitlow.

Primary and recurrent herpetic whitlow. Herpetic whitlow has been reported as a cross-infection among personnel working in clinics or hospitals. Vesicles or bullae suggest the diagnosis; confirmation is by virus isolation. Eight cases in medical personnel whose duties included contact with the oral cavity of infected patients were seen within a period of 3 years. The disease started as a paronychia in 3 instances, but involvement of the pulp space was the rule. Radiating pain along the C7 distribution was noted in 4 patients. Numbness and hypoesthesia following the acute episode were observed in one patient. Two individuals suffered recurrent attacks of herpetic whitlow at the same site.


Calcifying acne lesions.

Calcifying acne lesions. Cutaneous calcium deposition can occur in long-term inflammation. The deposition of calcium in long-standing acne may produce either small pigmented nodules or no visible lesions. The former type is or no visible lesions. The former type is much less common, with the nodules containing particulate spheres of mineral which probably represent true osteomas. Both types of calcification can be demonstrated roentgenographically and may occur in as many as half of all cases of severe long-standing acne.


Na+-K+ and HCO-3 ATPase activity in retina: dependence on calcium and sodium. With the use of homogenates of whole rat retina, the activities of Na+-K+-- and HCO-3-stimulated ATPases were measured in media containing different concentrations of calcium and sodium ions. In comparison to the Na+-K+ ATPase activity observed in the presence of 2 mM calcium and 130 mM sodium, the omission of calcium from the medium led to a 16-fold increase in activity. Furthermore, the omission of calcium and the reduction of the concentration of sodium in the medium to 25 mM also resulted in an increase in activity of ninefold. This study also demonstrates, for the first time, the existence of a HCO-3 stimulated. ATPase activity in the retina. These results are discussed in relation to recent observations on the effects of calcium- and bicarbonate-free media on the electrical and metabolic activities of the rat retina.


Transitional stages in the histochemical development of muscle fibres during post-natal growth. Serial frozen sections of longissimus dorsi muscles from seven pigs at different live weights (13 to 127 kg) were reacted for ATPase by the calcium method at an alkaline pH and for NADH oxidative activity. One hundred muscle fibres from each animal were identified individually in serial sections and their staining intensity was measured with a microscope photometer at 600 nm. For each section, staining intensity of fibres (% tranmission) was measured and converted to the nearest one-tenth unit of the range from the darkest to the lightest staining fibres. Frequency of occurrence of fibre types was plotted on a 10 X 10 grid using the range co-ordinates for NADH oxidative activity (vertical) and ATPase activity (horizontal). The commonly recognized histochemical fibre types in this muscle appeared as crowded areas in the grid but, in many cases, these areas were part of a continuous 'L' shaped range, a continuous but skewed distribution with regard to NADH oxidative activity was detected. In fibres with NADH oxidative activity of 0.6 to 1.0 units of the range, a continuous but irregular distribution with regard to ATPase activity was detected. Within this range, there was some evidence of a growth-related shift towards weaker ATPase activity.


A method for correlating ultrastructural and histochemical data from individual muscle fibers. The present study describes a new method making possible correlation of histochemical and ultrastructural data from identical muscle fibers. The method appears a valuable tool in studies where both light and electron microsocopy are required to obtain all necessary information.


Myosin light chain patterns of individual fast and slow-twitch fibres of rabbit muscles. Single muscle fibres were isolated by microdissection from freeze-dried samples of rabbit psoas and soleus muscles. The individual fibres were typed according to qualitative histochemical reactions for succinate dehydrogenase or NADH-tetrazolium reductase and for alkaline Ca2+-activated myofibrilla myosin ATPase after acid or alkaline preincubation. Methods are described for electrophoretic analysis by means of polyacrylamide disc electrophoresis in the presence of SDS of total myofibrilla proteins in single fibres after pre-extraction of soluble proteins. Fast-twitch white fibres revealed a myosin light chain pattern characteristic of "fast- type" myosin with three light chains of apparent molecular weights of 22,300 (LC1) 18,400 (LC2) and 16,000 (LC3). Fast-twitch red fibres were indistinguishable in this respect from fast-twist white fibres and showed an identical pattern of myosin light chains. Slow-twitch fibres could be characterized by a myosin light chain pattern typical of myosin of slow-twitch muscles with peptides of the apparent molecular weights of 23,500 (LC1Sa), 23,000 (LC1Sb) and 18,500 (LS2S). Slow-twitch fibres isolated from soleus as well as from psoas muscle were indistinguishable with regard to their myosin light chain patterns, thus suggesting that fibres of the same histochemical type correspond in their myosin light chain patterns irrespective of their origin from different muscles.


Some determinants of the reinforcing and punishing effects of timeout. Some determinants of the reinforcing and punishing properties of timeout were investigated in two experiments. Experiment I began as an attempt to reduce the frequency of tantrums in a 6-yr-old autistic girl by using timeout. Unexpectedly, the result was a substantial increase in the frequency of tantrums. Using a reversal design, subsequent manipulations showed that the opportunity to engage in self-stimulatory behavior during the timeout period was largely responsible for the increase in tantrums. Experiment II was initiated following the failure of timeout to reduce the spitting and self-injurious behavior of a 16-yr-old retarded boy. Using a multiple-baseline design, the nature of the timein environment was shown to be an important determinant of the effects of timeout. When the timein environment was "enriched", timeout was effective as a punisher. A conception of timeout in terms of the relative reinforcing properties of timein and timeout and their clinical implications are discussed.


Germination-defective mutant of Neurospora crassa that responds to siderophores. A conditionally germination-defective mutant of Neurospora crassa has been found to be partially curable by ferricrocin and other siderophores. The mutant conidia rapidly lose their membrane-bound siderophores when suspended in buffer or growth media. Germination is consequently delayed unless large numbers of conidia are present (positive population effect). This indicates that the mutant has a membrane defect involving the siderophore attachment site.


Isolation and preparation of pretyrosine, accumulated as a dead-end metabolite by Neurospora crassa. Pretyrosine is an amino acid intermediate of phenylalanine and/or tyrosine biosyntheses in a variety of organisms. A procedure for the isolation of high-quality pretyrosine as the barium salt is described. Stable solutions of ammonium pretyrosine that are suitable for use as substrate in enzyme assays can be prepared in good yield with relatively few purification steps. A triple mutant of Neurospora crassa, bearing genetic blocks corresponding to each initial enzyme step of the three pathway branchlets leading to the aromatic amino acids, accumulates prephenate and pretyrosine. Although the time courses of prephenate and pretyrosine accumulations were found to be parallel in any given experiment, the ratios of the two metabolites varied as much as 100-fold depending upon such variables as carbon source, temperature of growth, accumulation, and especially the presence of aromatic pathway metabolites. Under appropriate nutritional conditions of accumulation, pretyrosine concentrations in excess of 4 mM in culture supernatant fluids were obtained. Strains individually auxotrophic for phenylalanine or tyrosine accumulate lesser amounts of prephenate and pretyrosine. The metabolic blocks of the mutant result in high intracellular levels of prephenate, which is then partially transaminated to pretyrosine. In N. crassa, pretyrosine is a dead-end metabolite since it is not enzymatically converted to phenylalanine or tyrosine. At a mildly acidic pH, pretyrosine is quantitatively converted to phenylalanine in a nonenzymatic reaction.


Membrane asymmetry and expression of cell surface antigens of Micrococcus lysodeikticus established by crossed immunoelectrophoresis. Crossed immunoelectrophoresis of Triton X-100-solubilized plasma membranes of Micrococcus lysodeikticus established the presence of 27 discrete antigens. Individual antigens were identified as membrane components possessing enzyme activity by zymogram staining procedures and by reactivity of certain antigens with a selection of four lectins in the crossed-immunoelectrophoresis (immunoaffinoelectrophoresis) system. Absorption experiments with intact, stable protoplasts and isolated membranes established the asymmetric nature of the M. lysodeikticus plasma membranes. Of the 14 antigens with determinants accessible solely on the cytoplasmic face of the membrane, four possessed individual dehydrogenase activities, and a fifth was identifiable as a component possessing adenosine triphosphatase (EC 3.6.1.3) activity. Evidence from absorption studies with isolated membranes suggested that antigens such as the adenosine triphosphatase complex were more readily accessible to reaction with antibodies than was succinate dehydrogenase (EC 1.3.99.1), for example. Twelve antigens were located on the protoplast surface as determined by antibody absorption, and the succinylated lipomannan was identified as a major antigen. At least five other antigens possessed sugar residues that interacted with concanavalin A. With the antisera generated to isolated membranes, there was no evidence suggesting that any of these antigens was not detectable on either surface of the plasma membrane. From absorption experiments with washed, whole cells of M. lysodeikticus, it was concluded that the immunogens on the protoplast surface were also detectable on the surface of the intact cell. However, some of the components such as the succinylated lipomannan appeared to be exposed to a greater extent than others. The cytoplasmic fraction from M. lysodeikticus was used as an antigen source to generate antibodies, and 97 immunoprecipitates were resolvable by crossed immunoelectrophoresis. In the cytoplasm-anticytoplasm reference immunoelectrophoresis pattern of precipitates, three of the immunoprecipitates unique to the cytoplasmic fraction were identifiable by zymogram staining procedures as catalase (EC 1.11.1.6), isocitrate dehydrogenase (EC 1.1.1.42), and polynucleotide phosphorylase (EC 2.3.7.8). The identification of membrane and cytoplasmic antigens (including the above-mentioned enzymes) provides a sensitive analytical system for monitoring cross-contamination and antigen distribution in cellular fractions.


Generation of miniplasmids from copy number mutants of the R plasmid NR1. Small, closed circular deoxyribonucleic acid molecules, called miniplasmids, were observed in Escherichia coli harboring copy number mutants of the R plasmid NR1 after growth in medium containing tetracycline. The level of tetracycline resistance conferred by the copy mutant plasmids was lower (3 to 6 microgram/ml) than that conferred by NR1 (100 MICROGRAM/ML). The presence of the miniplasmid enhanced the level of tetracycline resistance conferred by the copy mutant. Miniplasmids of molecular weights 4 X 10(6) to 13 X 10(6) were found. They carried no antibiotic resistance markers and could be eliminated by growth in the presence of chloramphenicol and/or streptomycin-spectinomycin. Studies with the restriction endonucleases EcoRI and Sal I indicated that the miniplasmids are derived from the region of the copy mutant plasmids that contains the origin for replication of the resistance transfer factor. There were approximately 12 copies of the miniplasmid per chromosome, compared with 3 and 6 copies of the copy mutants of NR1. The miniplasmids appeared to be incompatible with the copy mutant plasmids.