Slow 514 nm absorption phases and oxygen exchange transients in Ulva. 1. The slow 514-nm spectral changes in Ulva were studied using bright continuous 650-nm light. Transient and steady-state absorption changes were compared with changes in net rate of O2 exchange in a system designed to measure both parameters simultaneously. 2. Time courses of the 514-nm absorption change show three phases following the onset of light: one rapid increase and two slower (larger than or equal to 1 s) transient increases. Upon cessation of the light three transient absorption phases also follow: a rapid decrease and two slower (greater than 1 s) transient increases. Parallel transient phases (but opposite in sign) were found at 480 nm. 3. The kinetics of the slow 514-nm absorption transients correlate with the characteristic induction transients in net O2 exchange. 4. Similar difference spectra and the restoration kinetics of the light-on and light -off transient phases indicate that the slow 514-nm absorption changes reflect the same component(s) and process(es). 5. The experimental results are discussed in terms of the electrochromic hypothesis for the 515-nm absorption shift. We interpret the slow 514-nm absorption changes in Ulva as a reflection of relatively slow ionic readjustments across the photosynthetic membranes.


Calcium binding to rabbit skeletal myosin under physiological conditions. At a free Mg2+ concentration of 1.0 mM, myosin binds one Ca2+ per molecule when the Ca2+ concentration is 20 muM, a value in the concentration range expected during contraction of skeletal muscle. Mg2+ alters Ca2+ binding in a complex manner, not by simple competition. In the range from 20 to 100 muM Mg2+ it produces positive cooperativity between the high-affinity Ca2+ binding sites, in addition to shifting binding to higher Ca2+ concentrations. High-affinity Ca2+ binding is not significantly affected by the addition of ATP, increase in ionic strength to 0.1 and changes in temperature. Ca2+ binding did not increase actin-activated ATPase activity in the absence of regulatory proteins, but rather inhibited it.


DNA replication in nuclei isolated from bovine lymphocytes. DNA replication was studied in nuclei isolated from phytohemagglutin-stimulated bovine lymphocytes. The mitogen treatment induced more than 60% of these cells to engage in DNA synthesis with a peak of synthetic activity occurring about 48 h after the addition of phytohemagglutinin. Throughout this response the ability of isolated nuclei and nuclear sonicates to synthesize DNA in vitro was proportional to the synthetic ability of the intact cells of origin. The subcellular systems appear to continue synthesis at replicative sites which were active in vivo. The rate of in vitro synthesis by both nuclei and nuclear sonicates was about two-thirds that of intact cells. The one cytoplasmic and two nuclear DNA polymerase activities separated from these cells were found to have properties similar to those of other eucaryotic cells.


A requirement for RNA, protein and DNA synthesis in the establishment of DNA replicase activity in synchronized HeLa cells. DNA replicase activity of synchronized cultures of HeLa cells was assayed by a permeable cell technique during normal S-phase and under conditions of restricted RNA, protein, or DNA synthesis. Inhibition studies with puromycin, cycloheximide, actinomycin D, and 2-mercapto-1-(beta-4-pyridethyl)benzimidazole revealed that the establishment as well as the maintenance of DNA replicase activity in S-phase cells was dependent on the continued synthesis of both RNA and protein. Measurements during limitation of DNA replication by hydroxyurea, cytosine arabinoside, or restricted availability of thymidine indicate that a low level of DNA synthesis is required to activate or assemble some subunits of DNA replicase. Evidence for the existence of short-lived RNA and protein factors essential for DNA replicase activity is discussed.


Characterization of the genome of Phycomyces blakesleeanus. DNA of Phycomyces blakesleeanus was extracted from whole mycelia and from nuclear and mitochondrial organelle fractions obtained from sporangiophores. DNA from all three sources exhibits one symmetrical band at p equals 1.688 g/ml in CsCl buoyant density gradients. Reassociation data are consistent with kinetic division of the DNA into three components: very rapidly renaturing (fraction I), rapidly reassociating (fraction II) and slowly reassociating (fraction III) base sequences. These components comprise approximately 10%, 20% and 64% of total cell DNA. Kinetic fractions were prepared from total cell DNA and reassociated separately. The corrected rate constant is 11.3 M-1-S-1 for the rapidly reassociating component and 0.055 M-1-S-1 for the slowly reassociating component. Based on these data and the data from unfractionated total cell DNA, the genome size of Phycomyces is approximately 1.9-10-10 daltons, 6.7 times that of Escherichia coli.


Size distribution and cell-free translation of globin-coding HnRNA from avian erythroblasts. Nuclear RNA from avian erythroblasts was isolated, fractionated by dimethylsulfoxide gradient centrifugation, and the fractions pooled according to increasing sedimentation values. Addition of 125I-labeled globin 9 S mRNA before centrifugation revealed a contamination of high molecular weight RNA by globin mRNA. However, the distribution of label was not identical to the distribution of globin synthesizing activity of unlabelled nuclear RNA when the RNA was translated in a cell-free system of Ehrlich ascites cells. Whereas more than 95% of 125I activity was found in the 0-20 S pool, only 33% of globin synthesizing activity of unlabelled nuclear RNA could be found in this pool. Most of the synthesizing activity was found in the 20-55 S pool and smaller amounts in even more rapidly sedimenting fractions. This conclusively demonstrates the existence of nuclear RNA with globin-coding sequences most probably representing the globin mRNA precursor. The precursor has a higher molecular weight than globin 9 S mRNA. Furthermore we could show that only a small percentage (smaller than 20%) of the proteins synthesized by addition of nuclear RNA from avian erythroblasts to the cell-free system represents globin chains. When 9 S mRNA is added to the cell-free system, more than 70% of the newly synthesized proteins are globin chains.


Poly (A)-containing polyribosomal RNA in sea urchin embryos: changes in proportion during development. Evidence is described which suggests that a large fraction of sea urchin embryo polyribosomal RNA is not polyadenylated, and that the proportion of poly(A)-containing RNA increases during embryological development. Cleavage, early blastula, and mesenchyme blastula stage embryos were treated with cordycepin, and the effects of the drug on polyribosome appearance were determined. Polyribosomes in cleaving embryos and early blastulae were unaffected by the drug. However, there was a significant reduction in the amount of polyribosomes present and in the amount of newly synthesized RNA entering polyribosomes in mesenchyme blastulae after cordycepin treatment. Polyribosomal RNA was isolated from cleavage, early blastula, and mesenchyme blastula stage embryos, and the RNA was characterized by sucrose gradient centrifugation. Cleaving embryo polyribosomal RNA has a prominent 9 S peak and a smaller 20 S peak, and the larger RNA is heterodisperse. There is no poly(A) associated with the 9 S and 20 S material, but there is poly(A) associated with the RNA sedimenting faster than 20 S. Early blastula and mesenchyme blastula polyribosomal RNA have 22 S and 26 S peaks, respectively. This later stage RNA is quite heterodisperse and poly(A) is associated with RNA in all size classes larger than 9 S. Finally, the proportion of polyadenylated light and heavy polyribosome RNA was determined for cleavage, early blastula, and mesenchyme blastula stage embryos. Approximately 20-25% of newly synthesized light polyribosome RNA is polyadenylated and this amount is relatively constant during early development. However, the proportion of newly synthesized RNA in heavy polyribosomes that is polyadenylated increases during development: 25% at cleavage, 33% at early blastula, and 48% at mesenchyme blastula stages.


Effect of colchicine on collagen, albumin and transferrin synthesis by cirrhotic rat liver slices. Collagen synthesis was found to be increased in liver slices of rats made cirrhotic by chronic administration of CCl4. The liver function was impaired, as determined by an increased retention of conjugated bilirubin and low serum albumin values. However, when animals received colchicine simultaneously with CCl4, collagen synthesis and deposition were inhibited, and the liver function appeared normal. When a group of rats was made cirrhotic by chronic administration of CCl4, and then kept for 30 days without further treatment, fibrosis persisted and collagen synthesis was very low. However, the liver function was severely impaired. When similar rats received L-azetidine-2-carboxylic acid during the 30-days period following CCl4 administration, there was a slight but not significant improvement in liver function. The collagen synthesis and the extent of fibrosis were similar to the controls. However, if similar rats received colchicine during the 30 days period, collagen synthesis was almost negligible, there was a slight decrease in fibrosis and there was a great improvement in liver function. In all the cirrhotic animals studied, transferrin biosynthesis remained constant.


Conformational studies on modified proteins and peptides. IX. Conformation and immunochemistry of hemoglobin reduced at some carboxyl groups by diborane. A derivative of apohemoglobin was prepared by reduction with diborane at -10 degrees C for 2 h. Four aspartate and two glutamate side chains were reduced per half molecule to their corresponding alcohols. The modifications were identified as: alpha asp 64, alpha-asp 74, alpha-asp 75 and alpha asp 85;beta-glu 121 and beta-glu 6 (or beta-glu 7). Recombination of reduced apohemoglobin with unmodified ferriheme yielded the corresponding reduced hemoglobin derivative which was electrophoretically homogeneous. Hemoglobin and reduced hemoglobin had different electrophoretic mobilities and showed slight spectral differences. Titration with ferriheme revealed that reduced globin possessed an anomalous heme-binding behavior. The s 20,w of reduced hemoglobin was slightly higher than that of hemoglobin. Agregation of the derivative was also apparent from its behavior in gel filtration. ORD and CD measurements indicated substantial conformational differences between hemoglobin and its derivative. Immunochemical studies showed that the derivative reacted poorly with antisera to hemoglobin and, conversely, hemoglobin showed little reaction with antisera to the derivative. The results are discussed in terms of the three-dimensional structure of hemoglobin in the crystalline state and its relationship to the structure in solution.


Aggregation of linear biopolymers induced by cooperative binding of ligands. Owing to "stacking" interactions certain positively charged dye molecules display biologically significant cooperative binding to single stranded polyanions. Temperature jump experiments with such systems reveal a peculiar slow relaxation process which cannot be ascribed to the actual binding reactions. In the special case of poly(glutamic acid) and acridine orange, light scattering measurements show it to be caused by a strong aggregation of the polymer-dye complexes depending primarily on the degree of binding. This effect is clearly reflected in changes of the visible absorption spectrum. Their wavelength dependences indicate that the aggregation is apparently associated with a spectral flattening as well as an enhanced binding of the dye which both result in a decrease of its absorbance. The thermodynamics and kinetics of the phenomenon can be understood on the basis of a simple model.


The influence of cadmium ions on the adsorption of prothrombin onto A1(OH)3 as a means to purify prothrombin. 1. In the presence of CdSO4(1mM),Al(OH)3(1.3% w/v) completely adsorbs the coagulation factors VII, IX, and X from normal plasma, but factor II (prothrombin) is adsorbed for about 50% only.2. A purification procedure for factor II is developed, using Al(OH)3 adsorption in the presence of Cd2+ as a first step and using column chromatography only once. A 750-fold purification is obtained at a 24% yield. 3. Comparison of the prothrombin thus obtained, with prothrombin isolated by the method of Kisiel and Hanahan (Biochim. Biophys. Acta (1973) 304, 103-113) does not show significant differences in amino acid composition, N-terminal amino acid, molecular weight or immunological properties. 4. Comparison of the two prothrombin preparations in a thrombin-generating system shows that although the final yield of thrombin from a given amount of prothrombin in both preparations is the same, the initial velocity of thrombin formation from our preparation is comparable to that of native prothrombin, whereas the other preparation is converted significantly slower.


Activation of a pro-enzyme by a stoichiometric reaction with another protein. The reaction between prothrombin and staphylocoagulase. The reaction between prothrombin and staphylocoagulase was investigated and the following conclusions were drawn: (a) Optimal amounts of the active reaction product (coagulase-thrombin) are found when equimolar amounts of prothrombin and staphylocoagulase are added together. (b) The molecular weight of coagulase-thrombin equals the sum of the molecular weights of staphylocoagulase and prothrombin when estimated both by gelfiltration and by sodijm dodecylsulphate-polyacrylamide gel electrophoresis. (c) The amino acid composition of coagulase-thrombin cannot be distinguished from the sum of the amino acid compositions of prothrombin and staphylocoagulasd. (d)in a preparation of coagulase-thrombin the N-terminal amino acids are those of prothrombin (alanin) and staphylocoagulase (aspartic acid). (e) An antibody against coagulase-thrombin precipitates prothrombin and staphylocoagulase but not thrombin. (f) We put forward the hypothesis that the thrombin activity in coagulasethrombin is the result of a stoichiometric reaction between one molecule of prothrombin and one molecule of staphylocoagulase, and limited proteolysis does not play a role in this mechanism.


A new principle in biospecific affinity chromatography used for purification of cobalamin-binding proteins. To avoid using protein-denaturing agents for desorption, when purifying cobalamin-binding protein by biospecific affinity chromatography, an affinity column has been prepared where cobalamin is attached through a temperature-labile linkage to insolubilized 3, 3'-diaminodipropylamine. On desorption the protein is obtained in solution saturated with cobalamin. The method has been used for purification of intrinsic factor and transcobalamin I.


Action of heparin on thrombin-antithrombin reaction. Thrombin partially purified from bovine plasma can be inactivated at 60 degress C. In the presence of 10 units of heparin the extent of inactivation decreases. When thrombin and heparin are mixed and incubated for 5 min at 0 degrees C before gel filtration on Sephadex G-200, thrombin with heparin is eluted prior to either thrombin or heparin laone. These data suggest a complex formation between thrombin and heparin. Immobilized heparin binds thrombin. The enzyme can be eluted with 0.05 M Tris-HCl buffer, pH 7.3, containing an ion mixture of Na+, K+ and Ca2+ at 73, 3 and 11 mM, respectively, at 0 degrees C and with 0.05 M Tris-HCl buffer, pH 7.3, containing 0.5 M NaCl at 20 degrees C. During the same chromatographic procedure, antithrombin-III (heparin cofactor) partially purified from human plasma is eluted with 0.05 M Tris-HCl buffer, pH 7.3, at 0 degrees C as well as 20 degrees C. Although, as described in the literature, heparin binds to antithrombin, our findings suggest another possibility, i.e. that the binding of heparin to thrombin induces a conformational change in the enzyme facilitating a complex formation between thrombin and antithrombin-III.


Comparison of the cyanogen bromide peptides of insoluble guinea-pig skin and scar collagen. Insoluble guinea-pig skin collagen and insoluble dermal scar collagen were cleaved with CNBr and the peptides derived from the alpha1-chain were separated by ion-exchange and molecular-sieve chromatography. Comparison of the peptides from scar collagen with those from skin collagen showed that the former contained more hydroxylysine. Separation of the CNBr peptides showed that this increase in hydroxylysine was found not only in the non-helical regions, but was also seen down the helical portion of the molecule. Separation of the peptides revealed the presence of more peptides in digests of skin collagen than those of scar, and these have been attributed to the presence of Type III collagen in skin, but no evidence was found for the presence of this Type III collagen in dermal scar tissue.


Haemoglobin G Norfolk: alpha 85 (F6) Asp leads to Asn. The characterisation of haemoglobin G Norfolk (alpha2 85 Asp leads to Asn 2) is described. This variant has been identified in the heterozygous state in three apparently unrelated English families of which two are resident in England and the other in Canada.


Association of the long-acting thyroid stimulator protector with the immunoglobulin G fraction of serum from patients with thyrotoxicosis. Both the long-acting thyroid stimulator and the long-acting thyroid stimulator protector are associated with the immunoglobulin G fraction of human serum proteins. Fractionation of globulins by (NH4)2SO4 precipitation and DEAE-cellulose chromatography showed that the protector activity was associated with subfractions of differing electrophoretic mobility. Attempts to concentrate the protector in immunochemically pure immunoglobulin G prepared in this way may fail because of loss of protector activity with other immunoglobulin G fractions. Gel filtration on Sephadex G-150 indicated that protector activity was associated with 7-S proteins.


Two new hemoglobins. Hemoglobin Alabama (beta39(C5)Gln leads to Lys) and hemoglobin Montgomery (alpha 48(CD 6) Leu leads to Arg). The amino acid substitutions in two new hemoglobins found by electrophoretic screening during a survey in Alabama have been determined by column chromatography and amino acid analyses of their tryptic peptides. They are hemoglobin Alabama (beta 39(C 5)Gln leads to Lys) and hemoglobin Montgomery (alpha 48(CD 6) Leu leads to Arg). No harmful symptoms have been attributed to the presence of either hemoglobin.


Proteins selectively released from water-extracted human erythrocyte membranes upon citraconylation or maleylation. Electrophoretic analysis and chromatographic fractionation. 1. Citraconylation or maleylation (carboxylation) of water-extracted human erythrocyte membranes (membrane residue) solubilized up to 20% (w/w) of the membrane residue protein, but less than 2% of the phospholipids. Amino acid analysis and polyacrylamide gel electrophoresis in the presence of dodecylsulfate showed that the solubilization was selective. 2. Polyacrylamide gel electrophoresis in the absence of detergent separated the solubilized acylated proteins into four well-separated major zones and two or three minor zones. Two of the major zones contained proteins of high molecular weight (spectrin components) and the other two contained components of intermediate molecular weight. 3. The solubilized components of intermediate molecular weight could be partially purified by hydroxyapatite chromatography. 4. Upon incubation at pH 5 and 36 degrees C of the unfractionated proteins solubilized by citraconylation some components became specifically degraded or dissociated into subunits.


Specific interaction of chromatin non-histone proteins with DNA. The interaction of different preparations of chromatin non-histone proteins of rat liver and thymus with homologous and heterologous DNA was studied. It is shown by the method of fixation of non-histone proteins-DNA complexes on nitrocellulose filters that: (1) all the non-histone proteins preparations studied form complexes with DNA in 0.02 M Tris--HCl (pH 7.5)--3 mM MgCl2; (2) the main part of non-histone proteins interacting with DNA binds to it non-specifically; (3) a small part of non-histone proteins interacts specifically with the homologous native DNA in 5 M urea; (4) both homologous and heterologous denatured DNA binds non-histone proteins more effectively than the native one; (5) the specific interaction of non-histone proteins with the homologous denatured DNA is observed both without urea and in its presence. The specific interaction of a small part of non-histone proteins with the homologous native and denatured DNA is also shown by the method of non-histone proteins chromatography on polyacrylamide--agarose columns containing DNA. The data obtained are discussed in the light of the possible non-histone proteins role in the specific regulation of the transcription process.


The state of messenger ribonucleic acid and ribosomes in the cytoplasm of ethionine-treated rat liver. The administration of ethionine to female rats causes breakdown of hepatic polysomes. The state of mRNA and monomeric ribosomes after the polysome dissociation was studied. The mRNA was selectively labeled with [14C] orotate after a low dose of actinomycin D. Sucrose density gradient centrifugation of Triton X-100-treated cytoplasm revealed an accumulation of heterodisperse radioactive material with very large S values. This material was converted to smaller S values with deoxycholate treatment and was extremely sensitive to mild ribonuclease treatment. Since this material was banded at around 1.43 g/cm3 in CsCl gradient centrifugation and contained RNA with a distribution of S values characteristic of polysomal mRNA, this material was identified as mRNA-containing ribonucleoprotein particles. The monomeric ribosomes were shown to be dissociated into subunits in the presence of 0.5 M KCl, indicating that these lacked nascent polypeptide chains. When the animals were recovered from the ethionine treatment by subsequent administration of adenine and methionine, the heterodisperse ribonucleoprotein particles and monomeric ribosomes appeared to be utilized for the reformation of polysomes.


Ion effects on protein-nucleic acid interactions: the disassembly of the 50-S ribosomal subunit from the halophilic bacterium, Halobacterium cutirubrum. The 50-S ribosomal subunits from the extreme halophilic bacterium, Halo-bacterium cutirubrum, stable structurally and functionally in concentrated salt solutions were subjected to ionic environments depleted in either K+ or Mg2+ or both. Under these conditions specific classes of proteins are released from the subunit along with the 5 S RNA. Two-dimensional electrophoretic analysis of the resultant split protein fractions indicate some mutually exclusive effects of specific ions on the binding of specific proteins to the 23 S RNA as well as on the retention of 5 S RNA within the ribosomal macrostructure.


Prediction of energy cost from peak heart rate in lower extremity amputees. Heart rate, which is an easily measurable biomechanical index, has been known to bear a definite relationship with the energy cost of any activity. An investigation undertaken on two groups of lower extremity rehabilitees and a group of normal, healthy, sedentary, adult males, when the subjects were administered with a battery of tests representing four common activities of daily living and working and a fifth test for exercise tolerance (as suitable), has confirmed that the relationship between peak heart rate (beats/min) and energy expenditure (kcal/min) is linear and can be represented by the equation E=0.068 PHR-4.59, thus making it possible to predict the energy cost of any activity from the peak heart rate value. This method is a more direct, simpler and more accurate one for the assessment of rehabilitation effectiveness in clinics than the more widely used direct measurement of energy cost by indirect calorimetry.


The role of a department of physiological (clinical) measurement in the UK National Health Service. The relatively new concept of a centralized department of physiological measurement, and the ways in which such a department interacts with related hospital departments within the framework of the National Health Service, are be borne in mind when setting up such a department are outlined and, finally, discussed. A variety of structural and functional considerations which are to the author describes his own department in the light of these considerations.


Use of postpheresis plasma to improve granulocyte yields for transfusion. Humoral factors which stimulate release of mature granulocytes from body reserves are presumed to be the mechanism through which high yields of granulocytes are obtained from donors by filtration leukopheresis. Postpheresis plasma (PPP) obtained 2 hr after leukopheresis, when infused into normal rats, induced a peak granulocytosis at 3 hr of 22,000/cu mm above controls. A substance in the nylon filters, which caused a peak granulocytosis at 4 hr of 7600/cu mm above controls, was eliminated by washing the filter with 30 volumes of saline. Injection of PPP obtained following leukopheresis with washed filters resulted in an 8000/cu mm increase in granulocytes. One milliliter of PPP given 1 hr before pheresis increased the granulocyte yield from 4.3 to 8.7 times 10-7 granulocytes in a 2-hr run. We conclude that (1) a humoral substance elaborated by the host during filtration leukopheresis induces a granulocytosis in the donor, (2) a substance in commercial leukopaks, which can be eliminated by vigorous washing of the filters, may be responsible in part for granulocytosis observed during leukopheresis, (3) PPP may be used to increase granulocyte yields in donors undergoing leukopheresis.


"Early-peak" carbon monoxide production in certain erythropoietic disorders. The "early-labeled" peak (ELP) of 14CO excretion following injection of glycine-2-14C was used to study erythropoiesis in a patient with sideroblastic anemia and in four subjects with myeloproliferative disorders. The ELP was greatly enlarged in all patients, as compared with a normal volunteer. The contour of the peaks from the hematologically abnormal subjects suggested the presence of increased erythroid heme degradation. In the patient with sideroblastic anemia, all hours of the early peak were significantly reduced after transfusion. This was interpreted to mean that even the earliest or "nonerythroid" phase of the peak is influenced by erythropoietic activity, at least under conditions of erythropoietic stress.


Heparin-induced thrombocytopenia: confirmation of diagnosis with in vitro methods. Profound thrombocytopenia developed in a patient during treatment with heparin for venous thrombosis. The platelet count increased toward normal when heparin administration was stopped, but fell abruptly when the drug was again given. Platelet aggregation occurred when heparin was added to the patient's platelet-rich plasma, or to normal platelets plus the patient's serum. This serum also effected release of 3H-serotonin from normal platelets. This pattern of aggregation was clearly different from that occasionally caused by heparin in a control population. The data is consistent with an effect of heparin on platelets, possibly mediated by on immune mechanism.


Short communication: possible association of newly absorbed serotonin with nonmetabolic, granule-located adenine nucleotides in human blood platelets. [3H]-adenine-labeled human platelets in plasma were incubated with or without nonradioactive serotonin. Release reaction was then induced by ADP, epinephrine, collagen, or thrombin. Platelets that had been incubated with serotonin released four times as much serotonin as platelets incubated without serotonin. The specific radioactivities of the ATP and ADP released to plasma during release reaction induced with all four inducers were the same in both systems. This shows that when serotonin is taken up by human platelets, it enters the compartment containing nonmetabolic, granula-stored ATP, and not the compartment with metabolic extragranular ATP. These results suggest that the mechanism of serotonin storage in human platelets is similar to that in other species investigated, i.e., rabbit, guinea pig, and pig.


Erythropoiesis in steel mutant mice: effects of erythropoietin in vitro. Adult SI/SI-d mutant mice have severe macrocytic, normochromic anemia. Moreover these animals are unresponsive to the stimulation of erythropoietin in vivo. By means of a bone marrow cell suspension culture system, the present investigation shows that in adult SI/SI-d marrow, there are cells capable of responding in vitro to erythropoietin in a normal fashion. Moreover, the erythropoietin present in SI/SI-d serum is biologically active in vitro without any prior biochemical modification. These observations support the suggestion that there is a defect in differentiation in the erythroid cell lines of SI/SI-d mice in vivo due to an abnormal hemopoietic microenvironment.


Recovery of normal hematopoietic tissue and tumor following chemotherapeutic injury from cyclophosphamide (CTX): comparative analysis of biochemical and clinical techniques. Simultaneous alterations in the incorporation of 3H-thymidine (3H-TdR) into DNA are induced by CTX in normal host target tissues and L1210 ascites tumor. The timing of suppression and recovery of these nucleoside incorporation alterations was similar at the three CTX doses studied, but some evidence for a dose-response effect was seen as the magnitude of suppression of DNA synthesis increased with increasing dosage. A differential pattern of suppression and recovery of 3H-TdR incorporation in malignant and normal host tissues was observed. The pattern of suppression and recovery of the peripheral white blood count and bone marrow (BM) cellularity, two frequently studied clinical parameters of hematopoietic recovery, were out of phase with the recovery of BM-DNA synthesis and failed to accurately reflect the sensitivity of the BM to subsequent chemotherapeutic injury. In contrast, drug schedules based on the differential recovery patterns of the host tissues and tumor, reflected by their 3H-TdR incorporation into DNA, both reduced toxicity to normal mice and increased the survival of tumor-bearing animals.


Radiosensitivities of DNA molecules in lymphocytes from the circulating blood of man. The radiosensitivities of DNA molecules from lymphocytes of human circulating blood were examined by alkaline sucrose gradient centrifugation. The single-strand breaks of DNA per rad, including the breaks formed under the present alkaline condition, were 1.2 plus or minus 0.1 breaks per 10-12 daltons DNA per rad. When the cells were transformed, the number of breaks was found to increase to 1.8 plus or minus 0.2 breaks per 10-12 daltons DNA per rad. The lymphocytes are capable of rejoining radiation-induced single-strand breaks of DNA. The rate of rejoining was dependent upon types of the suspending medium. The rate increased to ten times of that of the non-transformed cells upon transformation.


Sex hormones and the regulation of erythroid spleen colonies development of fetal liver origin. The development of erythroid colonies of fetal liver hemopoietic cell origin in adult irradiated polycythemic mice was studied. It was found that orchidectomy sharply reduced the number of erythroid colonies developed in the spleen of these polycythemic male recipients. Estrogen injection to the orchidectomized polycythemic recipient did not further decrease the number of erythroid colonies developed. It is concluded that the development of erythroid colonies of fetal liver origin in polycythemic male recipients is maintained mainly by testicular testosterone. The complete suppression of these colonies in female recipients does not seem to be a result of inhibition by estrogen.


Post-transfusion purpura: a heterogeneous syndrome. Three new patients with post-transfusion purpura (PTP) are described. As the manifestations in two differ significantly from those of previously reported cases, they serve to expand the definition of this syndrome. Although all 14 previously reported cases have occurred in Pl-A1-negative females, one of our patients was a Pl-A-negative male. Moreover, a female whose postrecovery platelets possessed the Pl-A1 antigen is described. Antiplatelet antibody activity was detected in all three patients by the 51Cr release test; in contrast, only one reacted in the complement (C) fixation assay. Serum obtained during the acute episode from the PlA1-positive patient reacted against platelets from four of 11 normals by C fixation and against platelets from 48 of 53 normals by 51Cr release, including five of nine Pl-A1-negative platelet samples. This case represents the first instance of PTP in which the platelet isoantibody was not specifically directed against the Pl-A1 antigen. These observations suggest that PTP may be a more heterogeneous disorder than previously realized.


Mechanisms for elevated fibrin/fibrinogen degradation products in acute experimental pulmonary embolism. The mechanism and significance of elevated levels of serum fibrin degradation products (FDP) in pulmonary embolism were investigated experimentally. Dogs were embolized with autologous blood clot-incorporating canine 125I-fibrin and were infused with either saline, heparin, or streptokinase. Serial measurements were made of total FDP by hemagglutination inhibition assay and of radioactive FDP. After saline, the peak level of total FDP was 323 mug/ml, but radioactive FDP was only 8 mug/ml. After heparin, these values were 44 and 11 mug/ml, respectively, and after streptokinase, 415 and 20 mug/ml. The results suggest that under these experimental conditions the elevated levels of FDP in pulmonary embolism are derived mainly from lysis of fibrin deposited after embolization rather than from lysis of the original embolus. Heparin inhibits both fibrin deposition and elevation of FDP levels after embolism.


A new method of measuring the deformability of the red cell membrane. Red cells moving in a stream of fluid can be arrested by means of a network of fibrin threads. Those cells which fold over a fibrin thread present to view a flat strap-like portion of membrane in which the strain can be easily observed and accurately measured both optically and by scanning electron microscopy. Preliminary results obtained by this method show elastic strains of nearly 300% and indicate that the stress required to produce these large strains is an order of magnitude greater than that reported by other methods.


Heparin-induced hyperplasia of lymphoid tissue. Experiments on newborn and sexually mature mice and rats showed that repeated injection of heparin leads to an increase both in the number of lymphocytes in the thymus and spleen and in the number of hematopoietic stem cells forming endogenous colonies in the spleen. The lymphoid tissue and the pool of colony-forming units are conjecturally under the regulatory influence of the adrenocortical hormones and of the product of the mast cells-heparin.


Role of corticofugal influences in mechanisms of formation of cortical bioelectrical activity. In experiments on cats anesthetized with a mixture of chloralose and pentobarbital, reversible cooling of the 1st sensomotor area (SM-1) unilaterally to between 25 and 23 degree C was accompanied by a simultaneous and generalized change in the activity of other parts of the cortex. These changes were expressed either as the appearance of slow high - amplitude activity or, on the other hand, by a marked decrease in amplitude of the EEG. Warming the cooled region of the cortex was followed by restoration of the normal EEG of the other parts of the cortex. These changes in electroencephalographic activity of the cortical areas are discussed in connection with the existence of synchronizing and desynchronizing subcortical brain structures.


Effect of vagus nerve stimulation on coagulability of the blood in cats. Electrical stimulation of the vagus nerve in cats leads to the development of hypercoagulation, accompanied by an increase in the thromboplastic activity of the blood. Increased fibrinoloysis under these conditions is evidently protective in character and aimed at dissolving the clots that may be formed in the blood stream as a result of the development of hypercoagulation.


Effect of membranous structures of the Kupffer cells on blood clotting. The effect of subcellular fractions of the endothelia kupffer cells on blood clotting was studied. Cells were obtained from the rabbit liver by magnetic fractionation and the subcellular structures were than isolated from them by differential centrifugation. The inner membranous structures were found to carry thromboplastic activity. No correlation was found between the blood-clotting potential of the intracellular structures of the endothelial cells and the level of their cathepsin C activity.


Investigation of a pain syndrome of spinal origin (on the concept of the generator mechanism of the pain syndrome). A pain syndrome was induced in rats by means of a microinjection of purified tetanus toxin into the posterior horns of gray matter of the lumbosacral segments of the spinal cord. The toxin was used as a means of disturbing inhibitory mechanisms. Investigation showed that a pain syndrome can be reproduced if afferent stimulation from the periphery is blocked (by division of the nerves of the hind limbs or division of the dorsal lumbosacral roots on the side of injection of the toxin). Under these conditions the latent period of onset of the syndrome was lengthened and the degree of its development weakened a little in the initial stages by comparison with animals with intact afferentation. In many animals with blocked afferentation from the hind limb general manifestations (restlessness, aggressiveness, crying, etc.) were accompanied by a localized response in the form of increased licking, biting, or even chewing the tissues of the deafferented limb at the site of projection of the pain (the phantom syndrome). In some animals only the general reaction was observed without localization of the pain (protopathic pain). In all cases the attacks of pain arose paroxysmally. In animals with intact limb innervation the zones of licking were trigger zones of facilitated induction of an attack of pain. Injection of glycine into the affected posterior horns of the spinal cord abolished the pain syndrome during the time of action of the glycine. It is concluded that the pain syndrome is based on the formation of a generator of pathologically intensified excitation, as a result of disturbance of inhibitory processes, in the system of neurons connected with pain sensation. These mechanisms are evidently those principally concerned in the pathogenesis of all pain syndromes.


Genesis of arrhythmias and mechanism of electrical defibrillation of the heart. Atrial arrhythmias were induced in experiments on dogs by electrical stimulation or by local application of aconitine and methacholine to the atrium. The action of the defibrillator discharge on these arrhythmias was studied. The defibrillator discharge abolished the arrhythmias maintained by the circus movement of the excitation wave over the atria but did not abolish sinus tachycardia or ectopic aconitine tachysystoles. The threshold of the defibrillating effect depends on the existence of micro-or macro-reentries. Thmechanism of defibrillation consists of excitation of the atrial myocardium with a consequent decrease in the pathway for the circulation of excitation to below the critical size for maintaining the circus movement of the excitation wave. The axtion of the defibrillator does not inihibit the automatism of the nomotopic and heterotopic cardiac pacemakers.


Possible role of slow sodium-calcium channels in the mechanism of changes in electrical and mechanical activity of guinea pig heart cells in local anaphylaxis (the isoptine effects). The local anaphylactic reaction and the effects of histamine during blocking of the slow sodium-calcium channels by isoptine were investigated in the spontaneously contracting oracle of the atrium of a guinea pig previously sensitized to egg albumin. Simultaneously with the intracellular recording of the potentials, isometric contractions of the preparation were recorded by means of a mechanotron. The investigation showed that egg albumin (0.2 mg/ml) and histamine (0.1 mg/ml) are antagonists of isoptine (2-16 mg/liter) as regards its effect on automatic contraction and duration of the action potential. It is postulated that this anaphylatic reaction is based on activation of the slow sodium-calcium channels in the surface membrane of the myocardial fibers.


Inhibitors of erythropoiesis in patients with aplastic anemia. An inhibitor of erythropoiesis was found in the urine of patients with aplastic anemia. The inhibitor was concentrated with kaolin and separated from erythropoietin by subsequent fractionation with alcohol and gel filtration on Sephadex G-100 and DEAE-Sephadex A-50. The inhibitor blocked the effect of erythropoietin (standard C) when injected into mice with hypoxic polycythemia 3 H before or along with exogenous erythropoietin; partial neurtralization of standard C also was observed on incubation with the inhibitor.


Functional and morphological adaptation of the gastric glands to disturbances of external pancreatic secretion and experimental reflux pancreatitis. The mucous membrane of the gastric fundus was investigated in dogs after interruption of the external pancreatic secretion and the formation of experimental reflux pancreatitis. A marked increase in the number of chief cells and a decrease in the number of parietal cells were found in the gastric fundal glands between 6 and 15-17 months after the operation. Meanwhile the RNA concentration in the chief cells was increased. These changes were constant throughout the period of investigation (until 20 months). The results are in full agreement changes in the secretory function of the stomach described previously under the same conditions and they can therefore be regarded as being compensatory or adaptive in character.


Disturbance of the urodynamics of the upper urinary tract as a result of hormonal imbalance. Considerable functional disturbances of the urodynamics, consisting of hypotonia, hypokinesia, and dyskinesia of the pelvis and upper third of the ureters were detected by television excretory pyeloscopy and motion-picture urography in female rabbits after prolonged administration of progesterone and estradiol dipropionate. These findings suggest a role of hormonal imbalance in the pathogenesis of disorders of the urodynamics that are the possible causes of development of pyelonephritis, nephrolithiasis, hydronephrosis, and other diseases of the kidneys in women.


Action of the mycotoxin of Fusarium sporotrichiella v. sporotrichioides on lysosomal membranes. The effect of the mycotoxin of Fusarium sporotrichiella v. sporotrichioides (sporofusarin) was studied in vitro on the total and nonsedimenting activity of eight lysosomal enzymes: acid ribonuclease, aryl sulfatases A and B, beta-glucuronidase, alpha- and beta-galactosidases, beta-glucosidase, beta-acetylglucosaminidase, and alpha-mannosidase. Incubation of a suspension of rat liver lysosomes with an aqueous solution of sporofusarin led to inhibition of the total activity of the membrane-bound lysosomal enzyme beta-glucosidase. In a dose of only 1.6 x 10-5 M sporofusarin caused a significant increase in the nonsedimenting activity of nearly all the enzymes; in a concentration of 1.6 x 10-3 M most of the enzymes of the lysosomal matrix (beta-glucuronidase, beta-galactosidase, aryl sulfatases A and B) were liberated almost completely into the supernatant, and nearly all the beta-glucosidase also was liberated. It is postulated that damage to the subcellular membranes is an important component of the toxic action of sporofusarin.


Oxidative phosphorylation in the skeletal muscles of rabbits during dinithrophenol hyperthermia. After intramuscular injection of 2,4-dinitrophenol (DNP) into rabbits in doses of 10,15, 25, and 30 mg/kg the degree of elevation of the body temperature, the phosphorylation in skeletal muscle homogenates were found o increase with the dose. In the course of dinitrophenol hyperthermia (following injection of 25 mg/kg DNP) the changes in body temperature and oxygen consumption of the animals followed a parallel course with the changes in the level of oxidative phosphorylation in the skeletal muscles.


Effect of insulin and glucose on the glucose consumption and glycogen synthesis by the rat diaphragm. Insulin activated both the uptake of glucose-C-14 from the medium and its incorporation into glycogen by the incubated diaphragm: the percentage of glucose-C-14 incorporated into glycogen increased with an increase in the dose of insulin. An increase in the concentration of glucose in the medium also caused this more rapid assimilation by the muscle tissue. However significant stimulation of glycogen synthesis was observed only in the presence of a high (unphysiological) glucose concentration in the incubation fluid; the percentage of glucose-C-14 incorporated into glycogen remained small and constant. It is postulated that the glucose-C-14 found in the case of substrate control is the result of exchange of the glucose residues of glycogen with intracellular glucose.


Detection of lactate dehydrogenase isoenzymes in single mammalian oocytes during cleavage by a micromodivication of disc electrophoresis. A micromodification of the method of disc-electrophoresis in glass capillary tubes is described. It can be used to study the protein composition and activity of lactate dehydrogenase isoenzymes in single rat and mouse oocytes during clevage.


Hyperglycemia and gluconeogenesis in the liver of mice with tumors. Gluconeogenesis, when sharply stimulated by exhaustion of the liver glycogen reserves, is one of the factors maintaining the normal blood sugar level in mice with tumors. Hyperglycemia induce by glucose leads to an increase in the liver glycogen content and a decrease in the intensity of gluconeogenesis in control mice with tumors. Only in the latter, however, does glycogen synthesis from noncarbohydrate compounds rise again steadily after the injections of glucose are discontinued.


Stimulation of adenylate deaminase activity by serotonin. After a single intraperitoneal injection of serotonin into rats adenylate deaminase activity in the mitochondrial fraction of the liver was stimulated. The conditions under which a twofold increase in the deamination of AMP occurred, after serotonin administration,were determined. Preliminary blocking of monamine oxidase activity did not prevent this effect of serotonin.


Comparative effect of amphetamine and caffeine on spontaneous activity of sensomotor cortical units and their responses to stimulation of the caudate nucleus. In acute experiments on anesthetized cats activity of sensomotor cortical units was recorded during low-frequency stimulation of the head of the caudate nucleus. Amphetamine (1 mg/kg), although not significantly affecting the spontaneous firing rate, increased the number of spontaneously active cortical units. Meanwhile inhibition of these units during stimulation of the caudate nucleus was weakened. Amphetamine had a stronger suppressive action on the inhibition arising from the ventro-lateral portion than from the dorso-medial portion of the head. In the case of caffeine the location of the stimulating electrodes was unimportant.


Correlation between the toxicity of chemical agents and their inhibitory activity on isolated mitochondria. Correlation was found between the concentrations of compounds inhibiting respiration of isolated mitochondria and the toxicity of the same substances for the whole organism. This correlation was observed for different intensities of action in vivo and for different pathways of entry of the toxic agent into the body. The relationship discovered applies to all inhibitors of tissue respiration irrespective of their point of application and of the molecular mechanism of their action.


Hypersensitivity of delayed type manifested during the development of allergic lesions of the myocardium. Changes in humoral and cellular responses were studied during the development of allergic lesions of the myocardium in rabbits. These responses,manifestations of hypersensitivity of delayed type, are connected with transformation of cells of the active mesenchyme, responsible for the process of immuogenesis, into lymhocytes and plasma cells. In allergic lesions of the myocardium hypersensitivity of delayed type is manifested as lympho-marcrophagal reaction in the intermuscular stroma of the heart, the thymus, and lymph glands and also as blast transformation of the lymphocytes. A complex series of disturbances of immunologic homeostasis arises, in which the primary components are  micronaglopahies, blast-transformation of the lymphoctes and activation of iysosomal enzymes.


Production of influenza antibodies by lymphocytes of the respiratory tract. Antibody production against influenza virus by lymphocytes of the human nasopharyneal tonsils and the tracheal wall of rats was investigated. Atibody-producing cells were found in preparations of lymphadenoid tissue removed in the postepidemic period. Intranasal immunization of rats with living influenza vaccine led to the accumulation of producer cells in the tracheal wall and of antibodies in the secretions of the respiratory tract. Reimmunization was followed by a well-marked secondary response. A characteristic feature of secretory immunity is the slower formation of producer cells and of antibodies than is observed in the system of general immunity.


Effect of the rhythm of cyclophosphamide administration on its antitumor action and the immune response of mice with methylcholanthrene sarcoma. The therapeutic effect of cyclophosphamide (CP) in relation to the dose and rhythum of its administration was studied in C57BL/6 mice with a methycholanthrene-induced sarcoma. The optimal scheme giving a complete cure of the experimental animals was 5 injections, each of  199 mg/kg CP at intervals of 10 days. The effect of the total dose of CP on humoral antitumor immunity was demonstrated: after injection  of 100 mg/kg CP the animals' serum lost its ability to stimulate tumor growth and acquired cytotoxic properties. The serum of animals receiving the total doses of CP above 400 mg/kg lost its effect on tumor growth. The doses of CP used did not affect cellular antitumor immunity (the cytotoxicity of the lymphocytes and resistance of the tumor to retransplantation.).


Proliferation of the glandular epithelium of the resected rat stomach. Between 45 and 50% of the wall of the gastric fundus was resected in adult male rats. By pulse labeling and 5 injections of thymidine-H-3 in the course of the 24 h an increase in the number of cells synthesizing DNA and the number of dividing cells of all types was found in the chief glands of the stomach during the first 3 days after resection. On the 5th-10th day the level of proliferation of all types of cells was lower than in intact animals except for the DNA-synthesizing parietal cells. The daily number ofDNA-synthesizing cells of all types 3-6 months after the operation had risen to the characteristic level for intact animals. The number of cells on the surface and pit epithelium and the number of mucous neck and parietal cells were increased. Partial reduction and dedifferentiation of the zymogenic cells were observed.


Effect of local negative pressure on reproductive function. Local negative pressure acting in the region of projection of the internal reproductive organs in rats widened and enlarged the network of blood vessels, thereby increasing the specific area of transcapillary exchange. Activation of ovulation was observed and the number of follicles and the number of young rats in the litter were increased. Local negative pressure had a positive action on the developing embryo, fetus, and newborn animal.


The "memory" of salivary gland cells.

The "memory" of salivary gland cells. In experiments on dogs with a chronic fistual of the submandibular salivary gland intravenous or subcutaneous injection of pilocarpine at intervals of a few days was accompanied by a gradual increase in the volume of saliva, reaching a maximum after 5 injections. Retention of the 'trace' of the action of pilocarpine was observed also in experiments on a dog with vago-sympathetic anastomosis and subsequent parasympathetic denervation of the gland.


Reaction of smooth muscle cells of blood vessels to an increase in functional load. Changes in the muscle tissue of the caudal vena cava of rats and its branches after disturbance of the blood drainage were studied by the methods of isotope biochemistry, autoradiography, immunomorphology, and cytophotometry.  Enlargement and polyploidy of the DNA-synthesizing muscle cells with activation of protein synthesis in them and the formation of intimal layers resembling the muscular layers were found in the main trunk of the vein which was dilated the most. Activation of DNA and protein synthesis in the less distended femoral vein was much less marked than in the main trunk. Activation of protein synthesis occurred after a delay compared with the activation of DNA synthesis but both processes were phases of development of hypertrophy of the muscle tissue of the vessel wall in response  to increased functional loading.


DNA synthesis in various cells of the exocrine part of the intact albino rat pancreas. A differential count was made of the DNA-synthesizing cells in various parts of the exocrine epithelium of the intact pancreas of sexually mature male albino rats sacrificed 1 h after a single injection of thymidine-H3. The acinar cells showed low proliferative activity (0.18 plus or minus 0.05%). The highest labeling index was found in the epithelium of the ducts. However, the lining membrane of the ducts consisted of a heteromorphic system of cells with a varying level of DNA synthesis. The labeling index of the nuclei of the centroacinar cells was 2.5 times higher than that of the acinar epithelium and amounted to 0.48 plus or minus 0.17%, whereas the epithelium of the intercalary ducts had an extremely low labeling index: 0.09 plus or minus 0.09%, compared with 0.27 plus or minus 0.09% for the intralobular ducts and 0.50 plus or minus 0.08% for the interlobular ducts.


The treatment of resistant malignant ascites by insertion of a peritoneo-atrial Holter valve. Holter valves have been used in 9 cases to drain ascites, and in 1 case of pleural effusion, into the superior vena cava. All these effusions were due to malignant disease. Three patients died before the results of the operation could be assessed and in 1 case ascites rapidly recurred. Five patients were relieved until death from their primary disease. One patient died 3 1/4 years after insertion of the valve, but ascites had returned--probably as a result of recurrent blockage of the valve by particulate matter.


Decreased incidence of indomethacin-induced gastric ulceration in rats by bile duct diversion. Gastric mucosal ulceration was produced in rats following intraperitoneal injection of indomethacin (40 mg/kg). The incidence of ulcerations was progressively reduced as the bile flow was diverted progressively distal to the ampulla of Vater. It is suggested that bile reflux into the stomach may play a part in the pathogenesis of indomethacin-induced gastric ulceration in rats.


The aetiology of hypocalcaemia in acute pancreatitis. Plasma calcitonin, glucagon and parathyroid hormone were measured in patients with acute pancreatitis. Plasma calcitonin was not detectable in 6 specimens obtained from the hypocalcaemic patients. Plasma glucagon values were similar in patients with acute pancreatitis and control subjects and were unrelated to hypocalcaemia, which was not even induced by glucagon infusion. High or rising parathyroid hormone levels were noted in association with hypo-and normocalcaemia, suggesting that parathyroid hormone rises and maintains plasma calcium within normal limits. Plasma parathyroid hormone was, however, undetectable in 8 patients with prolonged hypocalcaemia. Deficiency of parathyroid hormone due to its destruction by proteolytic enzymes or because of parathyroid gland exhaustion is suggested as the major factor inducing persistent hypocalcaemia in acute pancreatitis. Administration of parathyroid hormone should, therefore, be considered in patients with acute pancreatitis when hypocalcaemia does not respond to intravenous calcium therapy.


An appraisal of peritoneal lavage in the diagnosis of the acute abdomen. Forty-seven selected patients presenting with acute abdominal symptoms were investigated by examination of the fluid obtained by peritoneal lavage. In 34 patients this confirmed the clinical diagnosis, in 11 it was a more positive help in that it corrected an erroneous clinical diagnosis and in 2 the results of the investigation were misleading.


Abdominal lavage in blunt trauma.

Abdominal lavage in blunt trauma. A technique for abdominal lavage is described and was used to evaluate the abdomen of 671 multiple trauma victims. In 44 per cent (299 cases) there was a bloodstained return and these were regarded as positive. Patients with a positive result underwent exploratory laparotomy which revealed that 89 per cent had significant intra-abdominal trauma requiring a surgical procedure, 8 per cent had trauma which did not require any active surgical correction and 3 per cent had no abdnormal findings. Of all the lavages performed, there were 0-11 per cent false positive and 0-03 per cent false negative results. The value of the test in the context of multiple trauma is emphasized.


Extrinsic duodenal obstruction in infants and children. A series of 34 patients with extrinsic duodenal obstruction is described. The diagnosis and treatment are discussed. Five infants died and the findings are given. The remaining 29 have progressed normally.


Complete disappearance of popliteal arterial thromboses in 2 patients. Two young male patients with complete spontaneous resolution of popliteal thrombosis are described. The thromboses had been present for roughly 3 1/2 and 4 months. Such resolution has not been previously reported. The cause of the lesions is unexplained. It is suggested, should such a lesion be encountered, that the correct treatment is exploration to exclude adventitial cysts and popliteal entrapment, and simple evacuation of clot and anticoagulation.


Long term review of the results of rubber band ligation of haemorrhoids. A long term assessment (mean 4.8 years) of the value of rubber band ligation treatment for haemorrhoids has been obtained by a questionnaire answered by 125 of 147 patients. Eighty-nine per cent of patients were cured or satisfied with the results of treatment, although complete absence of symptoms was only obtained in 44 per cent. Further conservative treatment (rubber band ligation or manual dilatation) was required in 12 per cent and only 3 (2 per cent) have had a haemorrhoidectomy. Similar results were obtained for patients with the additional symptoms of pain and irritation, the presence of large skin tags and for the majority of third degree piles.


Trans-sphenoidal hypophysectomy in disseminated carcinoma of the breast. One hundred and eight patients with advanced carcinoma of the breast were treated by trans-sphenoidal hypophysectomy and followed up for at least 1 year. The results were assessed both by clinical response and objectively by survival following hypophysectomy, as expressed by the survival ratio. The two methods showed close correlation. The results show that there is a variation in response relating to menopausal status and age at diagnosis, subsequent hormonal manipulaton and physical condition at hypophysectomy. Younger patients whose carcinoma is diagnosed after menopause carry a good prognosis, yet the older premenopausal patients, especially if their clinical condition is poor, do not benefit. Easily available clinical information can be a guide to prognosis.


Raised intracranial pressure due to spinal tumours: 3 rare cases with a probable common mechanism. Three cases are reported of raised intracranial pressure associated with spinal tumours. The theories which have been advanced to explain this complication are discussed and it is suggested that in many of the reported cases the underlying mechanism may have been intracranial spread of the tumour and not a blockage of the cerebrospinal fluid pathways with proteinaceous deposits, as has been widely maintained. Autopsy revealed the former process to have operated in 2 of the 3 cases and it may have done so in the third. Two cases had unusual features in common: they were malignant schwannomas (a very rare spinal tumour) and they had both developed within the field of irradiation given for benign conditions many years previously.


Arteriographic exposition of the parathyroids. Arteriographic exposition of the parathyroids in 52 patients consecutively referred for primary hyperparthyroidism is reported. Thirty-nine patients under-went neck exploration. In 29 patients pathologic parathyroids were found. The arteriographic evaluation was based on observation of the course of the inferior thyroid artery and the presence of parenchymal opacification. Comparison of arteriography and surgical findings revealed that selective angiograpy of the inferior thyroid artery had a high sensitivity and specificity in the detection of pathologic parathyroids far exceeding that of a non-selective technique.


Peak expiratory flow in youths with varying cigarette smoking habits. Measurements of peak expiratory flow (P.E.F.) were done on 195 boys arriving at a detention centre, and again eight weeks later at the time of their discharge. During this time they took much physical exercise, and cigarette smoking and drug taking were not permitted. At the initial assessment there was an impairment of P.E.F. inversely proportional to the amount of cigarettes smoked which was statistically significant. There was a significant improvement in P.E.F. between reception and discharge in all groups of boys with varying smoking habits, all groups except the heavy smokers achieving near normal results after eight weeks. The heavier smokers improved more than the non-smokers or light smokers, but this difference was not statistically significant. Suggested reasons for this improvement are discussed.


Early discharge of infants of low birth weight: a prospective study. A total of 495 African infants of low birth weight were discharged from Harari Maternity Hospital, Salisbury, between October 1972 and September 1973. Criteria used for discharge were (a) no clinical evidence of disease, (b) satisfactory feeding by mouth (breast or bottle or both), and (c) stable temperature control under normal room conditions. Of the 495 babies 264 fulfilled these criteria when they weighed 1801-1900 g (group 1), 99 when they weighed 1901-2000 g (group 2), and 132 (group 3) when they weighed 2001-2500 g. The overall follow-up rate of those infants living in greater Salisbury was 85-5%, the health visitor playing an important contributory role in their progress, especially those in group 1. More than two clinic visits in the first four to five weeks after discharge were essential for continuing weight gain in groups 1 and 2 but not in group 3. The mean daily weight gain for all babies at the end of four to five weeks was 26 g. Readmission rates for babies in groups 1,2, and 3 were 9-5%, 1%, and 0-8%, respectively, the largest single cause for readmission being bronchopneumonia associated with hypothermia. Altogether 60% of the readmissions occurred during the two winter months (June and July). Hypothermia, associated with low environmental temperatures played a significant part in morbidity and mortality, and twins, particularly in group 1, had a mortality rate three times greater than singletons in the same group. In general, even in underdeveloped communities singleton babies born outside the winter months with reasonable clinic or home visiting facilities can be discharged at a weight of 1800 g or more.


Persistent primary enuresis: a urodynamic assessment. Videocystourethrography with synchronous pressure and flow-rate recordings has been carried out on 50 patients referred for the investigation of persistent primary enuresis. Urodynamic studies showed nocturnal enuresis to be associated mainly with normal detrusor function and nocturnal plus diurnal enuresis mainly with abnormal detrusor function. Evidence is presented which suggests that these two distinct types of enuresis occur de novo and do not overlap. Out of 18 of formerly enuretic male patients nine with abnormal detrusor function showed persistent nocturnal plus diurnal symptoms.


Measurement of fibrinogen-fibrin-related antigen in serum as aid to diagnosis of deep vein thrombosis in outpatients. A study of serum levels of fibrinogen-fibrin-related antigen (F.R.-antigen) in outpatients presenting with clinical features suggesting deep vein thrombosis was undertaken. A raised serum level of this antigen (greater than 12 mg/1) is strong evidence in favour of the diagnosis of deep vein thrombosis. It is virtually conclusive evidence if other known causes of a raised level of the antigen are absent. On the other hand, a normal serum level of F.R.-antigen does not exclude even extensive thrombosis, and other objective techniques are required to substantiate the diagnosis.


Follow-up of emergency ambulance calls in Nottingham: implications for coronary ambulance servie. Information about patients in ambulance service records has been linked to that in the patients' hospital records in an attempt to make the most efficient use of a special ambulance service for patients suspected of having heart attacks. During one week 248 emergency (999) calls for an ambulance were made by the public in the city of Nottingham. The quality of information given to the ambulance centre was poor, and all four patients eventually found to have had a myocardial infarction were described as having collapsed. A further study of patients who were also described as having collapsed has led to a system which allows an ambulance controller to send a "coronary ambulance" only in answer to those emergency calls where there is a reasonable possibility that the patient has had a heart attack.


Lymphocyte reactivity in pregnant women and newborn infants. The mitotic response to phytohaemagglutinin (PHA) was determined in lymphocytes of mothers and their newborn infants obtained at delivery and seven days later by measuring the rate of 125 I-idoxuridine uptake into DNA in lymphocytes cultured in their own plasma and after washing and resuspension in fetal bovine serum. There was no difference in the unstimulated counts of maternal lymphocytes taken at delivery, whether unwashed or washed, compared with those from nonpregnant controls. With PHA stimulation the mitotic response of the maternal lymphocytes cultured in their own plasma was reduced compared  with that of the control lymphocytes but washed maternal cells showed a similar response to the controls. These findings suggest that the reduced lymphocyte mitotic response to PHA in pregnancy is due to a plasma inhibitory factor This inhibition was not evident in maternal blood taken seven days after delivery. DNA synthesis in unstimulated cultures from newborn infants at birth and seven days after birth was greater than that in adult control cultures. With PHA stimulation the mitotic response of cord-blood lymphocytes cultured in their own plasma paralleled that of control lymphocytes but washed newborn cells showed a greater response. Thus plasma suppression similar to that observed in the mother seems also to affect infants at birth. This inhibition was not demonstrable in blood taken from infants of 7 days.


Depression of bone marrow colony formation in gold-induced neutropenia. Bone marrow culture in semi-solid agar was used to assess the proliferative activity and the response to sodium aurothiomalate of the myeloid precursor cells from patients during and after recovery from neutropenia associated with the use of this drug. Colony formation was reduced during the neutropenia and returned to normal after recovery. The rheumatoid process itself did not impair colony formation even in patients with Felty's syndrome. Sodium aurothiomalate inhibited colony formation by normal marrow in a dose-dependent manner. Bone marrow colonies from patients who had recovered from neutropenia induced by sodium aurothiomalate were not abnormally sensitive to the inhibitory effect of the drug in vitro. The metabolism of gold is probably altered in a small proportion of patients, which causes high local concentrations within the bone marrow leading directly to marrow depression.


Can colonic bacterial metabolites predispose to cholesterol gall stones? The cholesterol content of biliary lipids increased significantly when 16 healthy volunteers ingested deoxycholic acid (DC) for two weeks in a daily dose of 100-150 mg. Serum cholesterol also fell significantly to 88% of the baseline levels. Since DC is formed in the colon we suggest that populations in whom there is a high colonic absorption of bacterially metabolized cholate--that is, DC--have an increased predisposition to cholesterol gall stones.


Assessment of clinical competence using objective structured examination. To avoid many of the disadvantages of the traditional clinical examination we have introduced the structured clinical examination. In this students rotate round a series of stations in the hospital ward. At one station they are asked to carry out a procedure, such as take a history, undertake one aspect of physical examination, or interpret laboratory investigations in the light of a patient's problem, and at the next station they have to answer questions on the findings at the previous station and their interpretation. As they cannot go back to check on omissions multiple-choice questions have a minimal cueing effect. The students may be observed and scored at some stations by examiners using a check list. In the structured clinical examination the variables and complexity of the examination are more easily controlled, its aims can be more clearly defined, and more of the student's knowledge can be tested. The examination is more objective and a marking strategy can be decided in advance. The examination results in improved feed-back to students and staff.


Histochemical mapping of catecholamine neurons and fiber pathways in the pontine tegmentum of the dog. The distribution of catecholamine-containing neurons and fiber pathways were studied in the dog's pontine tegmentum by the Falck-Hillarp method. There are fundamental similarities between the dog and other mammals. Most catecholamine fluorescent cell bodies in the dog are located within the nucleus locus coeruleus and subcoeruleus in the pontine tegmentum. They are more diffuse and extensive than in the rat, monkey and human, and rather similar to those of the cat. In this study, catecholamine fiber pathways were clearly traced without using the lesion-degeneration technique. The dorsal pathway from the locus coeruleus consists of more compact bundles of non-varicose fibers than the rat, cat and monkey. It runs parallel to the mesencephalic tract of the trigeminal nerve in pons and midbrain. The ventral pathway courses as loosely arranged varicose fibers in the midreticular formation of the pons and sends axon collaterals to the caudal midbrain to form a dense network of varicose fibers. There are fluorescent plexi of fine varicose fibers at the ventromedial and dorsolateral aspect of the superior cerebellar peduncle. Cells of the medial and lateral parabrachial nucleus, in addition to those of locus coeruleus, send fluorescent fibers to this peduncle. No fluorescent fibers are found in other peduncles.


Subcellular fractionation of rat cerebellum: an electron microscopic and biochemical investigation. III. Isolation of large fragments of the cerebellar glomeruli. By a combination of differential and sucrose density gradient (both discontinuous and linear) centrifugation, large fragments of the cerebellar glomeruli were isolated in high purity from hand homogenised tissue. The final preparation contained only about 1% of the tissue protein, but over 90% of its volume was accounted for by the glomerulus particles. The ultrastructure of the glomerulus particles was well preserved. The enzyme profile was characteristic: the glomerulus particles were enriched in glutamate decarboxylase (GAD) activity (relative specific activity (RSA), 2.54), but the RSA of choline acetyltransferase (ChAc) was only 1.05. These findings are consistent with the view that GAD activity is very high in the inhibitory Golgi terminals, which occupy only a small fraction of the total volume of the particles, and acetylcholine may be a transmitter only in a relatively small fraction of the mossy fibre terminals. The glomerulus particles also contained a high concentration of succinate dehydrogenase (SDH) activity (RSA, 1.91), whereas the RSA of glutamate dehydrogenase (GDH) was only 1.15. The great asset of this preparation for future investigations is that it is composed almost exclusively from pre- and postsynaptic neuronal structures. Fractions containing neuropil fragments of non-glomerular origin were also obtained, but the profile of the estimated enzymes did not indicate unique characteristics.


Effects of litter size on protein, choline acetyltransferase (cat), and dopamine-beta-hydroxylase (DBH) of a mouse sympathetic ganglion. Litters of 5, 10 and 17 pups were formed at birth. At 7, 14, 21, 28 and 60 days after birth mice were weighed and the cervical superior ganglia were examined for total protein and for CAT and DBH activities. At each age, the parameters examined were fitted into regression lines on the reciprocal of the litter size (L.S.R.); a highly positive correlation was found at each age between body weight and L.S.R., with maximum differences at weaning (day 21) followed by partial recovery after ad libitum feeding until day 60. A positive correlation was also found for total ganglion protein at days 21 and 28, with recovery practically completed at day 60. For CAT activity a positive correlation with L.S.R. appeared at day 14, with increasing coefficient and significance in later periods, until day 60. The temporal pattern of DBH activity for the 5-pup litter showed a peak at day 21, followed by a 35% decrease at day 28, with a steady level until day 60. A similar trend was observed for the 10- and 17-pup litters, but the peak occurred at day 28. Significant differences were found as early as day 7 between the 5- and the 17-pup litters, the maximum differences in the 3 experimental conditions being found at weaning, with a high and significant correlation coefficient, which persisted at day 60. Thus, the biochemical development and maturation of both presynaptic cholinergic nerve terminals and postsynaptic adrenergic neurones was evidently affected by the itter size during the suckling period, with persistent effects.


The freezing lesion. III. The effects of diphenylhydantoin on potassium transport within nerve terminals from the primary foci. Possible mechanisms by which dephenylhydantoin (DPH) controls seizures were examined. The effects of intraperitoneal DPH on seizure discharges within epileptogenic freeze lesions were correlated with DPH action on in vitro potassium uptake within synaptosomes isolated from the same freeze foci. When in vivo DPH suppressed seizure discharges, it stimulated in vitro potassium uptake within synaptosomes incubated in a high-Kplus (10 mM) media. With 2-5 mM Naplus and 10mM Kplus, DPH stimulation of synaptosome potassium uptake was reversed by ouabain. With 50 mM Naplus and 10 mM Kplus, DPH stimulation of potassium uptake was not reversed by ouabain. In low-Kplus (0.2-5 mM) media, DPH did not affect potassium uptake even when sodium concentrations were varied at 10-100 mM. In sham-operated controls and in non-epileptogenic lesions, the effects of DPH on synaptosome potassium uptake were identical to those previously reported in normal brains. These results strongly suggest that DPH controls the epileptogenic state by stimulating potassium uptake within synaptic terminals. DPH controls the epileptogenic state by stimulating potassium uptake within synaptic terminals. DPH enhances synaptic potassium uptake by stimulating the (Naplus-Kplus) pump and a second potassium uptake process which is insensitive to ouabain.


Diversity of coding profiles of mechanoreceptors in glabrous skin of kittens. We examined stimulul-response (S-R) profiles of 35 single mechanoreceptive afferent units having small receptive fields in glabrous forepaw skin of 24 anesthetized domestic kittens. Single unit activity was recorded with tungsten microelectrodes from cervical dorsal root ganglia. The study was designed to be as quantitatively descriptive as possible. We indented each unit's receptive field with a broad battery of simple, carefully controlled stimuli whose major parameters, including amplitude, velocity, acceleration, duration, and interstimulus interval were systematically varied. Stimuli were delivered by a small probe driven by a feedback-controlled axial displacement generator. Single unit discharge data were analyzed by a variety of direct and derived measures including dot patterns, peristimulus histograms, instantaneous and mean instantaneous firing rates, tuning curves, thresholds for amplitude and velocity, adaptation rates, dynamic and static sensitivities, and others. We found that with respect to any of the S-R transactions examined, the properties of our sample of units were continuously and broadly distributed. Any one unit might exhibit either a slow or rapid rate of adaptation, or might superficially appear to preferentially code a single stimulus parameter such as amplitude or velocity. But when the entire range of responsiveness of units to the entire stimulus battery was surveyed by a variety of analytic techniques, we were unable to find any justifiable basis for designation of discrete categories of S-R profiles. Intermediate response types were always found, and in general, all units were both broadly tuned and capable of responding to integrals of several stimulus parameters, our data argue against the usefulness of evaluating a unit's S-R coding capabilities by means of a limited ste of stimulation of response analysis procedures.


6-hydroxydopa during development of central adrenergic neurons produces different long-term changes in rat brain noradrenaline. 6-hydroxydopa (6-OH-DOPA) administered to rats during their early development produces long-term modifications in the content of brain noradrenaline (NA) which have regional differences. An increase in brain stem NA is observed when the rats are exposed to the drug between the day 14 of gestation and the 9th postnatal day. When 6-OH-DOPA is injected subcutaneously on the 13th postnatal day or later, there is a decrease in brain stem NA. On the other hand, the content of NA in the telediencephalon is depleted for the first time in rats exposed to the drug during the day 16 of gestation, the decrease is more evident when the injection is done on days 17 or 18 and the effect is also marked when the drug is administered in the period between the day of birth and the 20th day of age. These results indicate that 6-OH-DOPA exerts different effects during the process of development and that the increase in brain stem NA is not solely dependent on the depletion produced in the forebrain because both phenomena are temporally dissociated. The adrenergic neurons injured by the drug, most probably respond in such a way that leads to an increase in brain stem NA only during the period in which they are under the influence of the factors controlling their physiologic development.


Effect of gonadal hormones on enzyme activities in brain and pituitary of male and female rats. Gonadectomized male and female rats were treated with equimolar doses of estradiol benzoate (EB) or testosterone pripionate (TP) daily for one week and enzyme activities were measured in the basomedial hypothalamus, corticomedial amygdala, and pituitary. In females, the hypothalamus showed estrogen-dependent increases in glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (ICDH), and malate dehydrogenase (MDH). Activities of ICDH and MDH were elevated in the amygdala. In the pituitary, estrogen administration resulted in increased levels of G6PDH, 6-phosphogluconate dehydrogenase (6PGDH), and lactic dehydrogenase (LDH). The estrogen antagonist, MER-25, effectively blocked estrogen-dependent increases in pituitary G6PDH and 6PGDH. Administration of TP did not result in changed enzyme levels. In males, treatment with EB and TP resulted in significant elevations in some but not all enzymes that were increased by EB in the female. Estrogen-dependent increases of activity in males were noted in pituitary G6PDH, 6PGDH, and LDH, in hypothalamic MDH, and in amygdaloid ICDH. Administration of TP led to increased levels of pituitary G6PDH, 6PGDH, LDH, ICDH, and MDH, hypothalamic ICDH and G6PDH, and amygdaloid MDH. The pattern of enzyme changes found in male and female brain and pituitary is discussed in relation to behavioral responses to gonadal hormones, nuclear uptake of gonadal hormones, and metabolism of androgen.


Effect of gonadal steroids on activities of monoamine oxidase and choline acetylase in rat brain. Gonadectomized male and female rats were treated with equimolar doses of estradiol benzoate (EB) and testosterone propionate (TP) daily for periods of 3 days to 1 week and activities of monoamine oxidase (MAO) and choline acetyltransferase (ChAc) were measured in the cortex, hippocampus, basomedial hypothalamus, corticomedial amygdala and medial preoptic areas. After hormone treatment, changes in enzyme activities were found in those brain regions where gonadal hormones are known to affect sexual behavior and/or gonadotropin release and which contain putative hormone receptor sites. More specifically, EB administration to females resulted in decreased activity of MAO in the corticomedial amygdala and basomedial hypothalamus and an elevation of ChAc activity in the medial preoptic area and corticomedial amygdala while TP administration did not alter enzyme levels in any brain region. In contrast, EB administration to castrated males was without significant effect on enzyme activities while TP administration resulted in increased activity of MAO and ChAc in the medial-preoptic area. The estrogen antagonist, MER-25, given concomitantly with EB, effectively blocked EB-dependent changes in both enzymes in ovariectomized female rats. EB treatment to hypophysectomized females led to similar enzymatic changes as in ovariectomized females in all areas except the basomedial hypothalamus. Estradiol added directly to the enzyme incubation medium did not result in altered enzyme activities. Results obtained are discussed in relation to sexual differentiation of the brain, metabolism of gonadal hormones, and possible mechanism of gonadal hormone regulation of enzyme activities.


Changes in proteolytic enzymes and proteins during maturation of the brain. (1) Changes during development in the levels of proteinases and peptidases were measured in brain homogenates. At all ages di- and tripeptidase levels were 7-15-fold higher than proteinase activity. (2) Cathepsin A and D and neutral proteinase activity first decreased (during the 5 days before birth) and then increased (primarily during the first 10 days after birth) in development. The total enzyme content per unit weight of brain did not change greatly after 10 days, although specific activity fell owing to an increase in protein in older animals. (3) The developmental pattern of activities or peptidases measured with Leu-Gly and Leu-Gly-Gly and of arylamidases measured with Arg- and Arg-Arg-beta-naphthylamides was similar to that of proteinases. Total and specific activities increased rapidly after birth; then total activity did not change and specific activity decreased. (4) The proteinase content of tissue fractions (nuclear and lysosomal-mitochondrial) similarly reached a maximal peak in the rapid growth phase of the brain. (5) The decrease of hydrolytic activity after 10 days of age seems to parallel a decrease in the rates of protein breakdown in vivo, showing parallel behavior with decreasing protein turnover. In contrast, during the first 10 days of life protein turnover and calculated rate of protein breakdown in vivo decrease while the level of hydrolytic enzymes increases.


Functional correlates of low molecular weight peptide synthesis in Aplysia neurons. An attempt was made to determine whether the synthesis of low molecular weight proteins by certain neurons of Aplysia could be correlated with either pacemaker activity or neurosecretion. Electrophysiological tests indicated that the spontaneous generation of action potentials in cells L8 and L9 is due to an endogenous pacemaker mechanism. Gel electrophoresis of proteins synthesized by these neurons showed no evidence of low molecular weight material. The non-neurosecretory interneuron L10 synthesizes a 12,000 dalton protein, whereas the silent neurosecretory cell L5 synthesizes a lower molecular weight peptide. Taken with previous findings, these results indicate that there is no correlation between pacemaker activity and the synthesis of these peptides. There is, however, a strict correlation between neurosecretory activity and the synthesis of proteins of molecular weights lower than 12,000 daltons.


Equivalent stimuli for stripes in rats with striate cortex ablations. The purpose of the experiment was to determine which features of a visual discrimination task were used by rats with total striate cortex ablations. The task was vertical black and white stripes versus grey matched in flux. Probe trials were inserted in which stripes of two different orientations or a speckled pattern were substituted for either the positive or negative stimulus. It was found that the destriated rats were normal in response to stripes of different orientations, but showed a deficit with the speckled pattern. A finer analysis suggests that the latter finding reflected an increased preference for the speckled pattern by the destriated group.


Hyperdipsia induced by bilateral destruction of the locus coeruleus in rats. Bilateral lesions in the locus coeruleus (LC) of rats induced a urinary disorder and hyperdipsia. Dilatation of the urinary bladder, urinary retention and hematuria with bleeding originating from the urinary bladder, occurred immediately after the lesioning and continued for 2-5 days. Water intake increased 4 days after the lesioning. This hyperdipsia persisted for at least 4 days and then gradually returned to the control level. Food intake decreased for the first 5 days, and then returned to the control level. Lesions in the ascending dorsal bundle (DB) originating from the LC also produced hyperdipsia, but not urinary disorder. Destruction of the ascending ventral vundle (VB) originating from the noradrenaline (NA) neurons in the medulla oblongata did not affect eating, drinking or urination. The LC- or DB-lesioning caused a significant reduction of NA in the whole forebrain except the hypothalamus, whereas VB-lesioning caused reduced NA in the hypothalamus. In LC-lesioned animals, no significant changes were observed in serum osmolarity, Na+, K+, albumin and glucose in serum, or in the excretion of urine in the water-loading test.


The cerebral blood distribution in dogs and cats. An anatomical and functional study. On the basis of corrosion preparations and of microsphere studies, the following characteristics of the canine and feline cerebral circulation were observed. (1) In cats, a greater part of the vertebral arterial blood goes to the brain and it is more specifically restricted to the ponto-medullary and cerebellar structures. These structures received approximately 3 times more microspheres in cats than in dogs. (2) In dogs, an important amount of vertebral blood goes to the neck muscles, and the intracranial vertebral blood supply is spread over a greater area of the brain, including the thalamo-hypothalamic and posterior cortical zone. (3) In cats the thalamo-hypothalamic area receives a greater amount of blood via the common carotid artery than in dogs. (4) In both animal species, the vascular connections between the left and right side of the brain are more extensive in the vertebral than in the carotid bed. However, for either vascular bed, a more important left to right transmission was found in the dog.


Intracranial pressure increase and changes in microcirculation of the pial and iridial vessels in correlation to EEG, ECG, and arterial blood pressure. Controlled intracranial pressure increase was produced in 120 female albino rats by infusing homologous blood and saline solution into the cisterna magna. The behaviour of the brain current, cardiovascular system, and microcirculation of the iridial and pial vessels immediately after pressure increase was examined simultaneously. A direct correlation could be demonstrated between the decrease in brain electrical potentials and the intracranial pressure applied at a time of maximal cardiovascular stimulation. Different pathological mechanisms have been put forward, one of these being identified as pial arteriolospasms. Acting together with other factors these spasms might be responsible for the rapid drop in electrical activity of the brain current potentials.


Amino acid incorporation into neurons and glia of guinea pigs with experimental allergic encephalomyelitis. After in vitro incubation of brain slices from guinea pigs in the intermediate (10 days postinduction) stage of experimental allergic encephalomyelitis (EAE), incorporation of [14C]leucine into protein was increased in a glial-enriched fraction. By the late (17-18 days postinduction) stage of the disease, when EAE symptoms were manifest, both the neuronal- and glial-enriched fractions showed increased specific activity of their total protein. SDS polyacrylamide gel electrophoresis showed that the increased leucine incorporation occurred in those proteins which labeled in a control material. After intraperitoneal injection of [3H]leucine the incorporated radioactivity was slightly increased in unfractionated brains from EAE animals. The neuronal-glial ratios for protein-bound radioactivity indicated that the increased incorporation resided mainly in the glial population.


Indicator media for microorganisms degrading chlorinated pesticides. A bromocresol purple liquid indicator medium and an eosin-methylene blue agar have been developed for the demonstration and isolation of microorganisms able to degrade the chlorinated herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D). Plates of the eosin-methylene blue agar indicate individual 2,4-D-degrading bacterial colonies. Both indicator systems show the production of acid, presumably hydrochloric, during degradation of the 2,4-D in the media. Concentrations of 2,4-D required to give an acid reaction in media with varying concentrations of yeast extract were determined; the production of about 0.24 mmol of hydrochloric acid seems necessary to counteract the buffering effect of 100 mg of metabolized yeast extract. Acid production from the herbicide, 4-chloro-2-methylphenoxyacetic acid(MCPA), which in the salt form could yield only small amounts of hydrochloric acid, was inconsistent. The two indicator media should be useful in investigations of the microbial degradation of other acid-yielding halogenated pesticides.


Influence of temperature and salt concentration on the growth of a facultatively halophilic "Micrococcus" sp. Growth characteristics of a facultatively halophilic strain of Micrococcus, capable of growth in medium containing no added NaCl and in medium containing 5.5 M NaCl, were studied over a range of salt concentrations and temperatures. Optimal growth occurred at 35C in a medium containing 1.0 M NaCl. In a medium containing 0.1 M NaCl, growth only occurred if the incubation temperature was less than 30C. No growth occurred if KCl, LiCl, NH4Cl, MgCl2, or sucrose was substituted for NaCl at 35C. A reexamination of bacteria previously described as moderate halophiles is suggested.


In vitro and in vivo interactions between Erwinia amylovora and related saprophytic bacteria. Under carefully controlled laboratory conditions, a highly virulent strain of Erwinia amylovora coinhabited susceptible host tissues with a yellow saprophytic bacterium, which was invariably isolated from fire blight infected trees, with or without producing symptoms of the disease depending on the status of a number of environmental factors, both climatic and physiological. In particular, variation of temperature and sucrose concentration determined, independently, the equilibrium of a readily reversible alternation of predominance of the two bacteria. It is suggested that E. amylovora may sometimes exist as an avirulent resident on the surface or within healthy host plants when environmental conditions favor growth of the yellow saprophyte rather than the pathogen. Such conditions, which are more likely to be obtained in midsummer and the fall, include temperature fall or rise below or above the optimum for E. amylovora, decreased humidity or diminution of sap flow, and increased sugar content in the host tissues.


Kinetics of Naplus-dependent amino acid transport using cells and membrane vesicles of a marine pseudomonad. Sodium ion is required for transport of L-alanine and alpha-aminoisobutyric acid (AIB) into cells and membrane vesicles of a marine pseudomonad. Initial rate data obtained at various Naplus and amino acid concentrations were tested for fit by least squares analysis to sequential and Ping-Pong equations. The sequential case is preferred statistically at the 99% confidence limit. Cotransport of AIB and Naplus in cells could not be detected, perhaps explained by efflux of Naplus shown to occur from these cells. The kinetic analysis is consistent with formation of a ternary complex involving Naplus and the amino acid.


The effect of growth environment on the chloroform-methanol and alkali-extractable cell wall and cytoplasm lipid levels of Mucor rouxii. The distribution of chloroform-methanol and alkali-extractable lipids in the cell walls of aerobically grown filamentous cells from Mucor rouxii has been determined. The results have been compared with the corresponding lipid composition of yeast-like cells from M. rouxii, which can be produced in two ways: by growth under anaerobic conditions and by aerobic growth in the presence of 0.22% phenethyl alcohol (PEA). It was observed that in most cases the crude cytoplasmic fraction contained higher levels of several lipids (i.e., squalene, sterols, triterpenes, and fatty acids) than did the corresponding cell walls. The cell walls did, however, contain both "free" (chloroform-methanol extractable) and "bound" (alkali extractable) lipids although the relative amounts were markedly dependent on the cell growth environment. The aerobically grown filamentous cell walls contained higher levels of squalene, sterols, triterpenes, and fatty acids than did aerobically grown yeast-like PEA-induced cell walls and there was also considerable variation in the "free"/"bound" ratios of the various lipid components. The lipid levels in both the cell walls and cytoplasm of the anaerobically grown cells were considerably lower than those of the cells grown under aerobic conditions. In addition, the differences in the growth environment were also reflected in the compositions of the individual lipid fractions from both the cell wall and the cytoplasm fraction.


The origin of fatty acids in the hydrocarbon-utilizing microorganism Mycobacterium vaccae. The fatty acid pattern in Mycobacterium vaccae strain JOB5 was examined after growth on n-alkanes (C14-C18), 1-alkenes (C14-C18), 2- or 3-methyl octadecane, and 8-heptadecene. It was evident that monoterminal oxidation of n-alkanes was followed by beta-oxidation and that both parent fatty acid and products of beta-oxidation were incorporated into cellular lipids. Radioactive experiments demonstrated that there was desaturation of long-chain fatty acids. There was no evidence of chain elongation. Growth on 1-alkenes resulted in the incorporation of fatty acids that were products of two primary modes of oxidation: (1) methyl group attack resulting in omega-unsaturated fatty acids and (2) double-bond attack resulting in the removal of one carbon from the substrate. Cells of strain JOB5 grown on 2- and 3-methyl octadecane contained the corresponding iso- or anteiso-fatty acids in significant quantity. Cells cultured on 8-heptadecene contained 8- and 9-heptadecenoic acids, 6- and 7-pentadecenoic acids, 9- and 10-methyl heptadecanoic acids, and 7- and 8-methyl pentadecanoic acids. Fatty acid composition (C13 to C19) was affected by substrate chain length and was additionally modified by cellular control mechanisms.


Theoretical studies on the coexistence of competing species under continuous-flow conditions. In continuous-flow environments, such as a chemostat, we find that, in theory, stable mixed populations of competing species obeying Monod/Michaelis-Menten growth kinetics (extended to more than one limiting substrate) cannot coexist unless the number of growth-limiting substrates is equal to or greater than the number of species. Additional restrictions on the relative values of the growth-yield constants and input-substrate concentrations must also be satisfied. Examples of the theoretical growth of mixed cultures after initial inoculation in stable and unstable cases are presented. The ecological consequences of these findings are briefly discussed.