Microbial biodegradation of cellophase.

Microbial biodegradation of cellophase. The microbial biodegradation of cellophane (U.C.B.--Division Sidac) was studied. Preliminary experiments with pure cultures of seven cellulolytic microorganisms (Aspergillus sp., Penicillium sp., Chaetomium crispatum, Ch. globosum, Sclerotium rolfsii and two actinomycetes) revealed that the substrate as such was very recalcitrant, probably due to the occurrence of insoluble coating agents. Therefore, mixed cultures of the above mentioned cellulolytic microorganisms were used as inoculum. The cellophane showed a slow microbial degradation which starts only after 37 days of incubation. This long lag-phase is due to the unaltered presence of the coating agents. However, when the coating agents are extracted with tetrahydrofuran, the biodegradation starts after 10 days, resulting in a biodegradation rate of 85% after 52 days of incubation and a protein content of 30%. The endproduct (30% protein, 60% soluble sugars, 10% residual substrate) will probably be useful as compost.


Kinetic studies on the formation of nitrosamines I. Formation of dimethylnitrosamine in aqeous solution of perchloric acid. The kinetics of nitrosation of dimethylamine (DMA) in aqueous perchloric acid solution have been studied using a differential spectrophotometric technique. The rate law is Initial rate = e[DMA]0 [nitrite]2 0 [H+]/(f + [H+])2 where [DMA]0 and [nitrite]0 represent initial stoichiometric concentrations. At 310.0 K and mu = 2.0 M, e = (2.2 +/- 0.2) X 10(-5) M-1 s-1 and f = (1.28 +/- 0.02) X 10(-3) M. The associated activation energy is 56 +/- 3 kJ mol-1. A clear inhibition of the nitrosation rate by ionic strength has been observed in which only the kinetic parameter (f) has an effective change. It is concluded that under the experimental conditions of this work only the dinitrogen trioxid is the effective carrier for the nitrosation.


Pyruvate and citrate metabolism in the muscle tissue of Ascaris lumbricoides. Aconitase and NAD linked isocitrate dehydrogenase were present in Ascaris lumbricoides muscle at only very low activities, whilst there were significant levels of citrate synthase, NADP linked isocitrate dehydrogenase, 2-oxoglutarate dehydrogenase and succinic thiokinase. Pyruvate dehydrogenase was present in A. lumbricoides muscle at levels comparable with mammalian tissues and results suggest that it is modulated via a phosphotransferase/phosphatase system. The tricarboxylic acid cycle intermediates, citrate, isocitrate and 2-oxoglutarate were all detected in freeze clamped muscle, but their steady state levels were considerably lower than those found in mammalian tissues.


Investigations into the free-living phase of the cat hookworm life cycle. Laboratory studies were undertaken to evaluate the role of environmental factors on the development of free-living stages of the cat hookworm Ancylostoma tubaeforme. An index of development calculated from the proportion of eggs that developed to the infective stage and the reciprocal of the development period, has suggested that the absence of 'high' temperatures (26 degrees - 30 degrees C) would significantly affect the L3 population in contaminated soils. There were, however, no significant differences in the size and lipid content of L3 that had developed at the different temperatures.


[Japanese methods of psychotherapy and their socio-cultural determinants. Impressions from Japan together with a report on the IV Congress of the International College of Psychosomatic Medicine, Sept. 5--9, 1977 in Kyoto]. Impressions of a congress-visiting-journey through the highly-industrialized far-east Japan are the background for description of specific Japanese Psychotherapeutic-technics f.i.: the Morita-therapy, Naikan-therapy and the Fasten-therapy. To the analytically trained Psychosomatic-specialist of western mentality those medical applications appear rather bewildering if not frightening. Their psychodynamic and their effectiveness become more understandable regarding the social-cultural conditions in modern Japan founded and grown on a tradition thousands of years old. Psychoanalysis can't prosper there. During the congress of the ICPM (International College of Psychosomatic Medicine) in Kyoto it became evident to the author by visits to clinics and above all by experience of typical Japanese travelling impressions, how much just psychotherapy and it's special drive-dynamical intentions are to a major degree depending on social-cultural development, the norms of value, the religion and the philosophy of a society.--Nippon is unique--strange and fascinating.


Factors affecting the microbial and chemical composition of silage. III. Effect of urea additions on maize silage. The effect of urea additions on the microbiological and chemical properties of silage, produced from young maize plants (Darawa stage), was studied. Urea treatments, i.e., 0.25%, 0.50%, 0.75%, and 1.00%, stimulated higher densities of the desired microorganisms than the control, while undesired organisms showed lower counts (proteolytic and saccharolytic anaerobes). Addition of 0.25 to 0.50% or urea resulted in the production of high quality silage with pleasant small and high nutritive value, as confirmed by the various microbiological and chemical analyses conducted. Higher levels (0.75 and 1.00%) of urea decreased the quality of the product.


Serum-albumin agar used as slant for the differentiation between Trichophyton rubrum and Trichophyton mentagrophytes: specific reg pigmentation of T. rubrum. Trichophyton rubrum strains freshly isolated from clinical material exhibited red pigment formation within 7 days at 26 degrees C on slant serum-albumin agar, initial pH 7.0. In Trichophyton mentagrophytes strains this phenomenon could not be observed. Since in the case of T. rubrum, yellow pigmentation will appear at an initial pH of 5.0, the use of initial pH values of 7.0 and 5.0 is recommended for diagnostic application of slant serum-albumin agar.


[Neurologic syndromes in myocardial infarct patients during various stages of treatment]. The clinical picture of myocardial infarctions in complicated and manifold. A clinico-physiological analysis of neurological disorders gives ground to consider that in myocardial infarctions the process of irritation involves the peripheral vegetative apparatus and spinal segmental structures, as well as the hypothalmostem structures with the reticular formation, which in its turn is accompanied by disturbances of cortico-subcortical interrelationships. In patients with myocardial infarctions the vegetative disorders are of a regional character and may be expressed in symptomatology, characteristic of the stimulation of sympathetic ganglions on the left side, mainly in the stellate node.


The action of cathepsin B and collagenolytic cathepsin in the degradation of collagen. Cathepsin B and collagenolytic cathepsin were obtained from bovine spleen and human placenta and identified as thiol proteinases. Both enzymes degraded insoluble fibrous collagen maximally at pH 3.5 and soluble monomeric collagen near pH 4.5. The response to activators and inhibitors was similar for both enzymes. Collagenolytic cathepsin was unable to degrade the synthetic substrates of cathepsin B and was also shown to differ in its physico-chemical properties. Minor differences were noted in the action of these cathepsins on insoluble fibrous collagen from different tissues. It was concluded that the rate and extent of the dissolution of fibrous collagen was determined by the number and location of the interchain cross-links, the amount of the associated non-collagenous components and the type of solvent ions, but not by the collagen phenotype.


Reduced post-natal rise of testosterone in plasma of cryptorchid infants. Plasma testosterone has been studied in 31 full-term male infants born with bilaterally undescended testes (14) or unilaterally undescended testis (17). From 10 to 89 days after birth, the post-natal testosterone rise was significantly lower in the 18 infants who remained cryptorchid at 4 months than in the 13 who underwent spontaneous testicular descensus and in the normal controls. Blunted post-natal Leydig cell secretion in cryptorchids may relate to a primary LH defect and could contribute to the impairment of both testicular descensus and maturation.


In vitro and in vivo studies on Leydig cell function in old rats. Young adult (3 months old) and old (26-28 months old) male Wistar rats were studied. Testicular weight was 1.66 g (range: 1.33--2.86) in the younger group and 1.62 g (range: 1.04--1.90) in the older group. The total Leydig cell volume as measured by a quantitative histometric method was significantly larger in the old animals (-x = 0.153 ml vs. -x = 0.089 ml). The testicular HCG binding capacity was 2.54 ng HCG per 100 mg tissue (range: 1.88--3.77) in the younger animals and 1.83 ng HCG per 100 mg tissue (range: 0.80--3.02) in the older ones (P less than 0.01). Plasma testosterone was on an average 242 ng/100 ml (range: 72--1162) in the young adult rats and 91 ng/100 ml (range: 23--277) in the older rats. Plasma LH was only slightly  (P less than 0.05) decreased and was 49 ng LH-RP-1/ml (range: 14--120) in the younger group and 40 ng LH-RP-1 ng/ml (range: 5--98) in the older. When the testicular tissue pieces were incubated with different doses of HCG or dibutyryl cAMP and testosterone production was measured, identical dose-response curves for old and young tissue were obtained. After in vitro incubation with a NADPH generating system, which under the conditions used was a much stronger stimulus for steroid biosynthesis than HCG or dibutyryl cAMP, the tissue of young rats produced about twice the amount of testosterone precursors (pregnenolone, progesterone, 17alpha-hydroxyprogesterone and androstenedione) than old tissue.


Influence of "lytic cocktail" on blood flow and oxygen consumption in the rat brain. The influence of a sedative dose of "lytic cocktail" (chlorpromazine, promethazine and pethidine) on cerebral blood flow (CBF) and oxygen consumjtion (CMRO2) was tested in artificially ventilated rats, maintained on either 70% N2 or 70% N2O. When given alone, the lytic cocktail had no significant effect on CBF or CMRO2. However, in the presence of nitrous oxide there was a 25% reduction in blood flow and oxygen consumption.


tcPCO2 electrode design, calibration and temperature gradient problems. Transcutaneous PCO2 electrodes habe been constructed and evaluated on adults and children. Glass pH and silver reference electrodes were used at 44--45 degrees C, with either circulating water and a copper jacket, or with internal electrical heating. The skin surface PCO2 at 44 degrees C is about 1.33 times PaCO2 plus 3 mmHg when measured with electrodes calibrated in gas at 44 degrees C. Three temperature effects combine in this ratio: Heating raises blood PCO2 4.5%/degrees C, skin metabolism adds about 3 mmHg, and the cooling of the electrode active surface by skin increases electrode reading. Response time to step changes of PaCO2 was about 3 min to 63%, of which 1.2 min was sensor delay, the remainder skin CO2 washout. It was found important to use ethylene glycol-water mixtures rather than water for electrolyte to avoid bubble generation and drift. Heat transfer through the pH glass electrode has been increased by enlarging the internal silver electrode to virtually fill the entire glass electrode. Time required for initial vasodilation and stabilization is similar to that of tcPO2 electrodes, and accuracy of determination of PaCO2 appears to be better than +/- mmHg.


Low impedance pH sensitive electrochemical devices that are potentially applicable to transcutaneous PCO2 measurements. Two cases of low impedance, non-glass membrane electrodes for pH measurement were evaluated: (I) Metal--metal oxide electrodes and (II) Reduction-oxidation electrodes. The fundamental cause of oxygen sensitivity of metal-metal oxide electrodes were examined and three approaches for its suppression were proposed. For the case of Sb--Sb2Ox electordes, oxygen sensitivity can be attenuated partially by cell loading, either directly across the reference electrode or indirectly across a third slave electrode. In a PO2 range of 8--54 kPa, more than 95% of the PO2  response can be suppressed by loading the cell emf to half of tis open-circuit value. The oxygen sensitivity also was observed to diminished by grinding the metal-metal oxide and pressing it under high pressure into a pellet electrode. Other metal-metal oxide electrodes that have promise in transcutaneous  measurement are the Pd-PdO2 electrodes. The redox electrodes are typified by the Quinhydrone electrode. A membrane Quinhydrome electrode showed a sensitivity of 56 mV/Decade at 37 degree C and no oxygen sensitivity up to 50 kPa and a drift of 1 mV/h over a 24-h period. However, the stability deteriorated over a long-term period.


The effects of isoproterenol on fetal oxygenation. Infusion of isoproterenol (1 microgram/kg/min, i.v.) into the anesthetized pregnant rhesus monkey near term consistently reduced fetal oxygenation, despite diminishing myometrial activity. The decline in pO2 of fetal arterial blood (mean = 4.3 +/- 2.3 mmHg S.D.) was accompanied by an increase in pCO2 tension (mean = 4.6 +/- 2.7 mmHg) and a decline in pH (mean = 0.04 +/- 0.02 S.D.). There was an increase in heart rate and a widening of pulse pressure in the mother and also in the adequately oxygenated fetus providing evidence that the agent crosses the placenta. The poorly oxygenated fetuses developed bradycardia and hypotension. Administration of isoproterenol directly to the fetus elicited similar changes in the composition of blood, and in blood pressure and heart rate, to those observed after administration of the agent to the mother.


Glaucoma treatment with timolol.

Glaucoma treatment with timolol. Thirty-two eyes from 19 patients with either capsular glaucoma, pigmentary glaucoma or primary open angle glaucoma were given topical timolol and followed through a 3--6 months period. Twelve eyes were previously out of control on full medication including pilocarpine, epinephrine and acetazolamide. Eight of these eyes could be adequately controlled throughout the follow-up period on topical timolol or a combination of timolol and pilocarpine, but without resorting to acetazolamide. One eye needed acetazolamide in combination with timolol, and three eyes had to be referred to surgery. Twelve eyes were adequately controlled either on pilocarpine or full medication, but could be equally well controlled on timolol alone, or timolol and pilocarpine in combination. Acetazolamide could be withheld from all of the six patients who used this drug. Eight eyes were previously untreated. They could all be adequately controlled either on timolol (5 eyes) or timolol and pilocarpine (3 eyes). The study indicates that the effect of timolol 0.5% X 2 may be stronger than the combined effect of epinephrine 1% X 2 and acetazolamide 500 mg a day. Topical timolol was well tolerated. No side reactions occurred and the pupillary size and reactions were not influenced by the timolol treatment.


Microcalorimetric study of the effects of cephalexin and cephaloridin on Escherichia coli and Staphylococcus aureus. The kinetics of the antibacterial actions of cephalexin and cephaloridin against a strain of Escherichia coli and a strain of Staphylococcus aureus were studied by a flow microcalorimeter. The heat production was related to the number of viable organisms (CFU ml-1), the pH, the optical density of the culture medium (OD540), and the morphology of the antibiotic-exposed bacteria. No heat effects could be registered when the number of CFU was below 10(4) ml-1. The addition of cephalexin, 2.5 microgram ml-1 (5 x MIC), to cultures of S. aureus caused a decrease in the heat production which was only roughly correlated with the number of CFU ml-1. This was also the case when 9.0 microgram ml-1 (2 x MIC) of this drug were added to cultures of E. coli. Two to three hours after the drugs had been added, no heat effects could be registered for the following 6--8 hours, after which an increase in the heat production again occurred. The MIC and MBC of the organisms isolated during this late heat increase were 8--40 times higher than those of the parent test organisms. A direct relation between drug concentration and response, i.e. heat effects produced, was found when increasing concentrations of cephalexin, i.e. 1.0 up to 50 micrograms ml-1 (2--100 x MIC) were added in the logarithmic growth phase to cultures of S. aureus. In ampoule calorimetric experiments, E. coli was cultured in a non-aerated, sealed growth vessel in the presence of cephalexin or cephaloridin in concentrations corresponding to 1/2 x MIC. The thermograms did not differ in shape, although the heat effects occurred somewhat later in the culture containing cephaloridin.


Studies on calcium uptake by myometrial microsomes with particular reference to the dependence on inorganic phosphate and oxalate. Ca uptake by microsomes isolated from non-pregnant rabbit myometrium was potentiated by both inorganic phosphate (Pi) and oxalate anions. Both Pi and oxalate had little effect on the initial rate of uptake but a pronounced effect on the capacity of Ca uptake measured after 20 min which was greater in the presence of oxalate than that of Pi (5 mM each). The presence or absence of sucrose in the uptake medium had a significant effect on oxalate-induced potentiation of Ca uptake but not on that potentiated by Pi or that measured in the absence of either potentiating anion. A part of Ca accumulated additionally under the influence of sucrose could be removed by washing microsomes with KCl. Another significant difference between the pontentiating effect of oxalate and Pi was observed when the pH of the incubation medium was varied. In the presence of oxalate the pH optimum was between 6.4--6.8, whereas that in its absence or in the presence of Pi the optimal pH was around 7.2. Reduction in pH from 7.2 to 6.8 along with the substitution of KCl by sucrose resulted in 3-fold increase in Ca uptake when oxalate was used as the potentiating anion. The results suggest that Ca is taken up by a different mechanism in the presence of oxalate than that in its absence or when oxalate anion is substituted with inorganic phosphate.


A controlled long-term study of flunitrazepam, nitrazepam and placebo, with special regard to withdrawal effects. The hypnotic effect of flunitrazepam (Ro 5-4200), nitrazepam and a placebo was studied in 117 outpatients using hypnotics for at least 3 months prior to the study. They obtained various neurotropic drugs and this and other treatments were unchanged throughout the trial period of 13 weeks. This consisted of 3 weeks on the previously used hypnotic, 3 weeks on a test drug (during the first of these a doubling of the dose was permitted if the initial dose of 1 mg flunitrazepam, 5 mg nitrazepam or one tablet of placebo was not satisfactory) and 4 weeks' observation after a request to stop medication with the test drug. The effects were evaluated every week by self-ratings. Also noted were: the frequency of dose increase after 1 week of the test period, number of drop-outs in the test period, and failure in the attempt to stop taking the test drug. A "psychological concentration test" was done, as was a follow-up interview. The self-ratings had a good reliability and showed that more patients experienced shorter sleep induction, longer sleep time, better sleep quality and a subjective feeling of having had a better rest with flunitrazepam than with either nitrazepam or placebo. There were no differences between the nitrazepam and the placebo groups. Tiredness was the most common side effect and appeared in the same frequency in all groups. The number of patients who increased the dose after 1 week's medication, as well as the number of drop-outs, was significantly higher in the nitrazepam and placebo groups than in the flunitrazepam group. There was no difference in the ability to discontinue the medication between the test groups or between groups having previously used different hypnotics. The "psychological concentration test" did not reveal any differences between groups. It was concluded that withdrawal of a hypnotic in chronic users was not facilitated by the use of a placebo. This was interpreted as due to a strong psychological dependence upon the hypnotics and their lack of pharmacological effects during long-term treatment.


Demonstration of enzymatic activity converting azathioprine to 6-mercaptopurine. The enzymatic conversion of azathioprine to 6-mercaptopurine was detected at pH 6.5 with rat liver supernatants, although the non-enzymatic reaction predominated at pH 7.0 and 7.5. Glutathione S-transferase may catalize this conversion. Activities of the enzyme in liver with both zathioprine and 1,2-dichloro-4-nitrobenzene as substrate decreased upon carbon tetrachloride-induced hepatic injury. These results may explain an ineffectiveness of azathioprine in patients with severe hepatic damage.


Transmitters for the afferent and efferent systems of the neostriatum and their possible interactions. The surgical denervation techniques have been widely employed as a tool to identify the neuronal origin of various neurotransmitters in the basal ganglia. Besides the well known nigrostriatal dopaminergic pathway, there is a high glutamic acid-containing corticostriatal connection and the cholinergic thalamostriatal projection. On the other hand, it is strongly suggested that GABA is the transmitter for the strionigral and striopallidal efferent systems. Furthermore, no evidence was found to support strionigral or striopallidal cholinergic pathways. From the 6-OHDA experiments, it is proposed that the 6-OHDA not only has a selective toxic effect on dopaminergic neurons, but also affects the cholinergic neurons in the striatum, and may provide further evidence for dopaminergic-cholinergic interaction in the striatum. Finally, GABA may modulate dopamine metabolism in the striatum.


Enzymic studies on glial and neuronal cells during myelination. The formation of ethanolamine plasmalogen from labelled 1-alkyl-2-acyl-sn-glycero-3-phosphorylethanolamine was studied in neurons and glial cells of the developing rat brain. It was found that the conversion of the ether to the enol-ether bond of the 1-alkyl moiety by the neuronal and glial desaturase system requires unsaturated fatty acids at the 2 position of the substrate. There is almost no difference between the activity of the neuronal and glial desaturase during the period of active myelination, whereas the neuronal cell fraction of the adult rats displays a threefold higher enzyme activity as compared to the glial cells. Evidence for the involvement of a microsomal electron transport system in the enzymic conversion of alkylacyl-glycero-3-phosphorylethanolamine to ethanolamine plasmalogen was obtained by using specific antibodies against NADH-cytochrome b5 reductase. Cytochrome b5 stimulated the biosynthesis of ethanolamine plasmalogen.


Interaction between PCO2 and plasma [HCO-3] in regulation of CSF [HCO-3] in respiratory alkalosis and metabolic acidosis. The results of studies presented may be interpreted in light of the dual contribution theory of regulation of CSF acid-base balance, indicating significant interaction between systemic and local CNS mechanisms in H+ homeostasis in the brain and CSF. Both systemic and local mechanisms are at work in H+ homeostasis in the brain and CSF and depending on the specific acid-base disorder or the duration of the imbalance one or the other of the two factors may be the dominant one in maintaining H+ levels in the "normal" range. These factors come into play only when there has been a change in central pH with the primary aim of bringing the CNS [H+] towards normal.


Conditions for production of thermosensitive attenuated influenza virus recombinants. Recombination and cross-reactivation between virulent influenza viruses and a cold-adapted thermosensitive vaccine strain regularly produced genetically stable attenuated recombinants, the selection of which was based on the thermosensitivity marker. This marker, correlating with the safety of the recombinants for man was inherited independently on the properties of the haemagglutinin and neuraminidase surface antigens. There was no relationship between the thermosensitivity of the resulting recombinants and the decrease in the optimal temperature of the neuraminidase activity (OTNA marker). This indicates a separate localization in the viral genome of the mutation damages responsible for the expression of ts and OTNA markers.


Interspecies interactions of arboviruses. III. Competition for virus envelope antigens in mixed Getah and Sindbis virus populations. Interactions of Getah and Sindbis virus populations in mixed infections of 5 tissue systems were competitive or neutral. The type of interaction is regulated by the host cell and virus population density. On reproduction in mouse brains and SPEV cell cultures the mixed population exhibited antigenic markers of Getah virus; in Syrian hamster kidney and chick embryo cell cultures those of Sindbis virus; and in Chinese hamster kidney cell cultures the antigenic markers of both viruses. Mixed populations with monospecific antigenic characteristics contained genomes of both viruses detectable by the TC marker (dual spetrum of cytopathogenicity). Clonal analysis of such populations confirmed the occurrence of genome masking and demonstrated the formation of genotypically mixed particles. Possible mechanisms of the antigenic dominance of one virus and selective blocking of functions of the other virus are discussed.


Experimental pathogenesis of non-lethal herpesvirus infection and the establishment of latency. In rabbits inoculated into the right scarified cornea with the Kupka strain of human Herpesvirus type 1 (HHV 1), healing of the keratitis occurred between days 8--12 post infection (p.i.). Viral particles were seen in nuclei and cytoplasm of corneal epithelium cells and in a few fibrocytes of corneal stroma. By immunofluorescence and electron microscopy, the virus was demonstrated in single neurons and satellite cells of the homolateral Gasserian ganglion since 24 hr p.i. From 20 hr to 8 days p.i., HHV 1 was found in ganglia of 7 out of 15 rabbits, while the explantation was positive in 10 out of 12 animals. In the course of latency, single HHV antigen-containing cells were seen in ganglia of 3 out of 7 rabbits and virus isolation attempts before explantation always were negative. Viral antigen was found in single ganglion cells only in 20% of rabbits with healed corneal lesions while the explanation of the ganglion tissue yielded HHV at an unchanged high frequency (92% of animals).


Antiviral activity of dipyridamole derivatives. Among 46 novel pyrimido [5.4-d] pyrimidine derivatives, 26 compounds were found to exhibit antiviral activity as revealed in a test programme against Mengo, Coxsackie B1, fowl plague, vaccinia and pseudorabies viruses, as concerns inhibition of plaque formation and of infectious virus yield. Attempts to disclose structure-activity relationships by discriminant analysis pointed to a possible importance of hydrophobic substitution for the antiviral effectiveness against Mengo virus of the derivatives investigated.


Isolation of influenza virus from wild ducks (Anas platyrhynchos). In the course of studies on influenza virus ecology, Influenzavirus A was isolated from a cloacal smear from a wild duck (Anas platyrhynchos) caught in west Slovakia. The strain was identified as A(Hav7Nav1). The results of virus isolation experiments from other species of aquatic and other small birds were negative.


Influence of mild acid hydrolysis on the antigenic properties of phase I Coxiella burnetii. Mild acid hydrolysis of phase I Coxiella burnetii cells gradually cleaved off polysaccharide chains, which contain determinants of antigen 1, thus unmasking determinants of the diagnostically important antigen 2. This artificial phase I to phase II conversion was reflected by changed serological activity in the complement-fixation (CF) test and by changed nonspecific phagocytosis. The product of mild acid hydrolysis can serve as a complex diagnostic preparation, because it reacts in the CF test with antibodies directed to both C. burnetii antigens.


Immunological properties of the lipopolysaccharide-protein complex of Coxiella burnetii. Purified lipopolysaccharide-protein complex (LPS-PC) extracted by trichloroacetic acid from phase I Coxiella burnetii organisms induced in mice and rabbits fair levels of antibodies directed to antigen 1 and antigen 2, as detected by complement-fixation (CF), microagglutination (MA), opsonization-phagocytosis (OP) and serum protection (SP) tests. In guinea pigs only very low levels of MA antibodies against antigen 2 were demonstrated. In rabbit serum, MA antibodies directed to antigen 2 were found exclusively in the IgM fraction after the primary immunizing dose; the second dose was followed by gradual shift of MA antibodies to the IgG class. Two immunizing doses of the LPS-PC were more effective when testing antibody response in mice or protection of mice and guinea pigs against phase I virulent challenge.


Cytomegalovirus infections in the human population. Diagnostic and clinical experience gained in the period from 1966--1977 is summarized. Cytomegalovirus was isolated from 35 sucklings and 5 infants aged more than 1 year as well as from necropsy materials from 10 sucklings. The most frequently involved were the respiratory tract (77.8%), liver (53.5%) and haematopoetic apparatus (42.2%). Congenital defects were found in 22.2%. The incidence in the population varied from 20% in sucklings to 50--60% in persons aged over 40 years. The incidence of infection was higher in sucklings and infants in nurseries. Out of 37 adult patients with kidney transplantation, the virus was isolated from 10 (27%) and complement-fixing antibodies were found in 34 (91%) patients.


Strain differences in electrophoretic mobility of influenzavirus A polypeptides. Slab polyacrylamide gel electrophoresis of Influenza-virus A strains varying in antigenic composition of the envelope and in the natural host species showed the nucleprotein and membrane proteins to have a standard molecular weight of 58,000 and 24,000 daltons, respectively. The electrophoretic mobility and molecular weights of the light and heavy chains of haemagglutinin varied in the strains under study. The differences observed persisted on passaging and were transferred upon genome recombination.


Isolation of encephalomyocarditis virus from the pheasant (Phasianus colchicus) in Czechoslovakia. The first isolation of encephalomyocarditis (EMC) virus from the blood and brain of a pheasant (Phasianus colchicus) captured in Moravia is reported. Complement-fixation (CF) tests with reference antisera revealed no relationship to any of a wide range of known arboviruses.


Sensitivity of formation of adenovirus specific tumour antigen to leukocyte interferon. The effect of various concentrations of chicken leukocyte interferon was examined on the expression of adenovirus tumour (T) antigen in chick embryo cells. The inhibition of T antigen formation was dependent on the multiplicity of infection. The reduction of T antigen-producing cells was greater when interferon was applied before and after virus infection.


Rates of infection in, and transmission of, African horse-sickness virus by Aedes aegypti mosquitoes. Very low infection rates (less than 3%) were obtained when Aedes aegypti mosquitoes ingested blood contained 5.8--6.5 log10 MLD50/0.02 ml African horse sickness virus (AHSV). When A. aegypti mosquitoes were inoculated intrathoracically with virus, however, high infection rates were achieved. Mosquitoes infected by inoculum failed to transmit virus to embryonated hens eggs by bite, and virus could not be detected in membrane or blood when inoculated mosquitoes were allowed to engorge on uninfected blood through a chick skin membrane. It was concluded that the mosquito A. aegypti is unlikely to be an effective vector of AHSV.


Rickettsiella phytoseiuli and virus-like particles in Phytosfiulus persimilis (Gamasoidea: Phytoseiidae) mites. Electron microscope investigation of tissue of Phytoseiulus persimilis mites revealed its double infection with Rickettsiella phytoseiuli and virus-like particles. The presence of rickettsiae in tissues of the mite seemed to be decisive for establishment of virus infection.


Cross reactions among flaviviruses in macrophage migration inhibition assay. Adult Swiss albino mice were immunized intravenously with flaviviviruses. Cellular immune response was studied by macrophage migration inhibition assay. Migration of cells from sensitized mice was significantly inhibited in the presence of homologous virus or antigen. Migration inhibition was also significant with serologically related viruses, thus establishing cross reaction in cellular immune response.


Methodical aspects of the experimental chemotherapy of viral infections. An analysis of the methods of testing the antiviral activity in vitro (in cell cultures) and in vivo (in experimental virus infections) is presented. The necessity of applying two-stage screening in vitro is emphasized. The chemotherapeutic viral models is in vitro testing as well as the principles of modelling of experimental viral infections are examined. Criteria for stage-by-stage assessment of the antiviral activity of the substances tested are formulated.


The theoretical implications of chlorpromazine as a sensory integrative theory. In order to evaluate the influence of antipsychotic drugs used in the treatment of schizophrenia on therapy based on sensory integrative theory, this study, developed through library research, explores the theoretical bases for these two modes of treatment. Studies of chlorpromazine, a prototypical antipsychotic drug, show that its local action on the neurotransmitters of the brain may explain in theory its therapeutic efficacy. By citing evidence of sensory processing deficits in schizophrenic patients, a theoretical basis for the use of a sensory integrative approach to therapy is established. A comparison of these theories leads to the conclusion that a sensory integrative approach can serve as an important reinforcer of the therapeutic actions of chlorpromazine. Implications for treatment and research are considered.


Arterial hypoxemia following the administration of sublingual nitroglycerin. The effect of nitroglycerin on arterial blood gases and cardiovascular hemodynamics were studied in patients with coronary artery disease. In 13 premedicated patients blood gases and cardiovascular hemodynamics were studied before and 10 minutes after sublingual nitroglycerin (0.6 mg). In eight unpremedicated patients only blood gases were determined before and 10 minutes after sublingual nitroglycerin 0.6 mg. All studies were performed before induction of anesthesia with the patients in the supine position breathing room air. In both groups, arterial PO2 decreased significantly (p less than 0.001); pH and pCO2 did not change. In the 13 patients on which hemodynamic studies were performed, the mean arterial pressure  (p less than 0.001), cardiac index (p less than 0.001), central venous pressure (p less than 0.001), pulmonary artery (p less than 0.001) and pulmonary artery wedge pressure (p less than 0.001) decreased. Calculated values for systemic and pulmonary vascular resistance were not significantly altered (p greater than 0.4). This study gives conclusive evidence that nitroglycerin reduces arterial PO2 in most patients with coronary artery disease breathing room air in the supine position. The possible mechanisms and clinical implications are discussed.


Evaluation of the counterimmunoelectrophoretic (CIE) procedure in a clinical laboratory setting. The counterimmunoelectrophoretic (CIE) procedure was evaluated under clinical laboratory conditions to determine its validity and comparability with culture methods. The procedure was further evaluated to determine applicability to a variety of clinical samples. An inexpensive set-up was developed to utilize the CIE procedure at bench level. Results indicated the procedure to be sensitive in detecting Haemophilus influenzae, type b, and Neisseria meningitidis (meningococcus), group B. The procedure was more sensitive for detection of H. influenzae, type b, than for meningococcus, group B. The authors have confirmed the usefulness of the CIE procedure in the detection of group B streptococci, pneumococci and teichoic acid antibody to Staphylococcus aureus. Detection of Escherichia coli K 1 antigen was also accomplished by CIE. In the authors' laboratory the CIE procedure was superior to culture methods when used for the detection of H. influenzae, type b, and meningococcus group B.


Analysis of hydroxyzine hydrochloride, meperidine hydrochloride and atropine sulfate in glass and plastic syringes. The apparent stability of combinations of hydroxyzine hydrochloride and meperidine hydrochloride (50 mg/2 ml each) and of these two drugs (50 mg/2.5 ml each) and atropine sulfate (0.4 mg/2.5 ml) in prefilled glass and plastic syringes was studied. Syringes (3 ml) containing the combinations were stored at 25 C and 3 C for 10 days and analyzed at specific time intervals. Absorption spectra, chromatographic characteristics and pH were determined in addition to visual inspection. Results of these qualitative tests indicated that the mixtures apparently were stable for 10 days at room temperature or when refrigerated. No differences were found between solutions stored in glass and those stored in plastic syringes. Degradation of the syringe contents or appearance of additional constituents was not detected in any of the admixtures, and they were considered to be chemically compatible within the limitations of the study. The study suggests that storage of these combinations in syringes is feasible but the results cannot be extrapolated to drug solutions or syringes other than those studied.


Education and integration of midlevel health-care practitioners in obstetrics and gynecology: experience of a training program in Washington state. The impetus for the development of the Gynecorps Training Program was the identification of medically underserved populations of women in Washington State and their need for preventive health care, the maldistribution of physicians, and the success of a pilot program for training midlevel personnel conducted in 1972 by the Department of Obstetrics and Gynecology, University of Washington School of Medicine. The program trains community-based and -sponsored Registered Nurses and Physician's Assistants as women's health-care specialists, with the goal of delivering preventive care in obstetrics and gynecology to populations of high-risk, low-income, and/or underserved women. A summary of the training program, its curriculum, and the integration of its graduates into community health teams is presented and discussed.


Amino acid transport in diaphragms from newborn rats: evidence for insulin resistance. Diaphragms from rats under 24-h-old did not show the well-known increased transport of alpha-aminoisobutyrate found in older tissues in respone to insulin in vitro. A small effect was apparent by 3 days, and stimulation increased as donor rats aged (up to 4--5 wk). One-day diaphragms also had greater uptake than older tissues, due to both decreased Km and elevated Vmax. The change in insulin sensitivity did not result from alteration in the transport system used by alpha-aminoisobutyrate because uptake showed characteristics of the A system at both 1 day and older. Results suggest instead that the 1-day tissues had been made insulin-resistant by high insulin levels in donor animals. Plasma insulin levels of 1-day-old rats were 5 times those of 5-day animals. Elevating the plasma insulin levels of 5-day or 25- to 35-day rats led to a decreased effectiveness of insulin in vitro in stimulating alpha-aminoisobutyrate transport into their diaphragms. In the older animals, the stimulation was inversely proportional to the plasma insulin level 2 h after insulin injection.


Characterization of distal hydrogen ion secretion in acute respiratory alkalosis. The effect of acute respiratory alkalosis (ARA) on distal nephron H+ secretion was evaluated by measuring urine-to-blood (U-B) Pco2 in dogs with highly alkaline urine (urine pH greater than 7.8). ARA led to a significant decrease in U-B Pco2 and in urine HCO3 concentration; urine pH, however, increased significantly, indicating that the decrease in urine Pco2 was of greater magnitude than the decrease in urine HCO3 concentration. For any given urine HCO3 concentration urine Pco2 was lower (i.e., urine pH was higher) in ARA than in controls. Administration of tris(hydroxymethyl)aminomethane (Tris) during ARA resulted in a significant increase in U-B Pco2 to control values. In animals with moderately alkaline urine (urine pH 6.4--7.4) and high urine PO4 concentration, ARA resulted in a significant decrease in UB-Pco2 and urine PO4 concentrations. Neutral PO4 infusion in these dogs resulted in an increase in urine PO4 concentration and U-B Pco2 to control levels. These data demonstrate that ARA results in a significant decrease in U-B Pco2 that is not solely attributable to changes in urine HCO3 concentration. The observation that Tris and PO4 infusion during ARA raises U-B Pco2 to control levels suggests that the ability to secrete H+ is intact.


Differences in inducibility by glucocorticoids of rat liver TO and TAT. The modulation of tryptophan-oxygenase (TO) and tyrosine amino-transferase (TAT) in the rat liver after a single dose of hydrocortisone has been studied under various physiological conditions. Differences in the induction behavior of the two enzymes have been observed dependent on sex, age, amount of administered hormone, and presence or absence of the adrenals. Some of the results observed are the following: 1) TO reached its maximum induction level 3 h prior to TAT in normal male rats. This difference disappeared when adrenalectomized male rats or when female animals were tested. 2) The maximal induction values of both enzymes were 50--80% higher in adrenalectomized rats than in normal rats. This effect was independent of sex. 3) Higher doses of hydrocortisone were necessary for optimal induction of TO and TAT in normal than in adrenalectomized rats. 4) The minimum dose of hydrocortisone necessary for enzyme induction was significantly lower for TO than for TAT. 5) Actinomycin D caused a complete inhibition of the induction of TO and TAT when given simultaneously with the glucocorticoid. The inhibition was less complete the longer the interval between hormone and actinomycin D administration. The activity of TAT was suppressed to a larger extent than TO.


Ammonia metabolism.

Ammonia metabolism. The pathways responsible for an the mechanisms underlying the adaptive increase in ammonia production in response to acidosis are considered. It seems unlikely that the cytosolic pathways (glutamine synthetase, glutaminase II, phosphate-independent glutaminase, and gamma-glutamyl transferase) are of primary importance in the adaptive process, but the role of the purine nucleotide cycle has not been resolved. The intramitochondrially located phosphate-dependent glutaminase pathway is generally believed to be of primary importance. Adaptation involved either enhanced glutamine entry into the mitrochondria and/or activation of phosphate-dependent glutaminase, but the relative importance of each has not been resolved definitively. The overall adaptive response is probably modulated by factors regulating alpha-ketoglutarate metabolism to phosphoenolpyruvate, and possibly also by metabolism of TCA cycle intermediates. It seems unlikely that a decrease in systemic pH is the direct effector for the acidosis-induced increase in ammonia formation; however, the resulting decrease in urine pH may play a critical role. Other potential messengers, including potassium, glucocorticoids, mineralocorticoids, cyclic AMP, and calcium probably do not serve a primary function, but the importance of other circulating factor(s) is unclear.


Changes in the plasma anion gap during chronic metabolic acid-base disturbances. A basic premise in the utilization of the plasma anion gap in the assessment of acid-base disorders is that this parameter remains constant during hyperchloremic metabolic acidosis and metabolic alkalosis. Experimental data under in vitro conditions, however, cast serious doubt on this premise. The purpose of the present study was to characterize the plasma anion gap, estimated as (Na + K) - Cl + HCO3), in two large groups of dogs with graded degrees of chronic, HCl-induced metabolic acidosis or chronic, diuretic-induced metabolic alkalosis. The data indicate that the plasma anion gap decreases significantly in HCl acidosis and increases significantly in metabolic alkalosis; the predicted mean anion gap in animals with a plasma bicarbonate concentration of 10, 21 (normal), and 40 meq/liter approximated 13, 18, and 26 meq/liter, respectively. The observed variation in the plasma anion gap is interpreted as originating mainly from directional changes in the net negative charge of plasma proteins; these changes result from the titration process secondary to the altered plasma acidity and, in the case of metabolic alkalosis, from the additional effect of an increased plasma protein concentration.


Relationship between blood pH and potassium and phosphorus during acute metabolic acidosis. Metabolic acidosis is known to be associated with increased blood potassium and phosphorus concentrations but the influence of mineral versus nonmineral acids on these variables remains undefined. Therefore, we infused anesthetized mongrel dogs with 0.45% saline (controls), the mineral acids HCl and NH4Cl, and the nonmineral acids lactic, beta-hydroxybutyric and methyl malonic for 1-3 h. Administration of both mineral acids was associated with significant increases in plasma potassium. In contrast, infusion of the three monmineral acids did not result in increases in plasma potassium; in fact, the levels decreased initially in the majority of the dogs. Phosphorus concentrations were increased by lactic and beta-hydroxybutric acids, were unchanged by NH4Cl and HCl, and were decreased by methyl malonic acid. Although the mechanisms responsible for these changes remain to be elucidated, the findings indicate that short-duration infusion of mineral and nonmineral acids has substantially different effects on plasma concentrations of these predominantly intracellular ions.


Marijuana use in schizophrenia: a clear hazard. The use of marijuana as the independent variable produced a serious exacerbation of a psychotic process in four schizophrenic patients whose illness was otherwise well controlled with antipsychotic medication. Each patient served as his own control--the sole substance abused was marijuana and antipsychotic medication intake remained constant. Each time marijuana use at moderate levels began, there was exacerbation and deterioration. The author suggests that marijuana use is a special hazard to schizophrenic patients and that physicians should alert such high-risk patients to the possible untoward interaction between their illness and marijuana.


Current medication use and symptoms of depression in a general population. In a community survey, 41.8% of 771 men and 60.2% of 1,059 women reported having used one or more medications in the 48 hours before the interview. The use of medications and the number of medications used increased progressively with age among both men and women. Respondents who used four or more medications included significantly more high scores on a depression checklist than those who used fewer medications. The group of women who used minor tranquilizers and sedatives included significantly more high depression scorers than those who did not. Among both sexes, those scoring in the depressed range who were receiving psychotropic medication tended to be taking minor tranquilizers or sedatives.


Psychotropic drugs, summer heat and humidity, and hyperpyrexia: a danger restated. The authors note an apparent unawareness of, as well as a paucity of recent literature pertaining to, hyperpyrexia as a complication of psychotropic drug treatment. They report a case of hyperpyrexia in a patient receiving psychotropic agents during a summer heat wave; massive muscle destruction (rhabdomyolysis) with myoglobinuria and acute renal failure make this case of particular interest. Causative mechanisms, preventive measures, and the benefits of prompt recognition and treatment of this condition are discussed.


Simple, effective treatment of agoraphobia. For effective treatment of agoraphobia both patient and therapist must understand the mechanisms of sensitization and self-desensitization. Rather than aiming to adapt to difficult situations, to achieve desensitization by suggestions, or to avoid panic, the agoraphobe must learn to pass through panic and to rid oneself of drug dependency. This method of self-desensitization will, as a rule, achieve results quickly and does not necessarily depend upon finding the cause of the original sensitization.


Changes of glycosaminoglycan synthesis during in vitro ageing of human fibroblasts (WI-38). Synthesis rates of glycosaminoglycans by WI-38 cultures (diploid, human fibroblasts exhibiting a limited number of population doublings in vitro) were determined by incorporation of 35S-sulfate of 14C-glucosamine into cellular and extracellular glycosaminoglycans at different passage levels before phase out. A progressive decline in the synthesis of cellular and extracellular glycosaminoglycans occured during the last (about 4) population doublings. 35S-sulfate incorporation into extracellular glycosaminoglycans appeared to be somewhat more reduced than 14-C-glucosamine incorporation during the last passages. Analysis of the distribution pattern of incorporated label into various glycosaminoglycan types (hyaluronic acid, chondroitin sulfate, dermatan sulfate and possibly heparan sulfate) revealed an age-related relatively stonger decline of 14C-glucosamine incorporation into cellular and extracellular hyaluronic acid and of 35S-sulfate into extracellular chondroitin sulfate in comparison with the other glycosaminoglycan types. Addition of exogenous glycosaminoglycans (chondroitin-4-sulfate, chondroitin-6-sulfate, dermatan sulfate, hyaluronic acid, heparan sulfate, heparin) at 100 microgram/ml to the culture media during the last 7 to 10 population doublings before phase out did not increase the total number of population-doublings. Heparin exhibited a significant growth inhibitory effect at 100 or 500 microgram/ml. The changes in glycosaminoglycan metabolism are interpreted as an expression of cellular ageing, and such an in vitro system offers a model for analyzing the factors involved in or causing the induction respectively prevention of this functional change.


Experimental evidence for a unifying concept of the molecular mechanisms of the cellular aging and the cellular neoplastic transformation. Experimental data supporting a Unifying Concept of the molecular mechanisms of cellular aging and cellular neoplastic transformation of dividing cells in mass culture and clonal culture systems will be described. The Unifying Concept combines the hitherto antithetically presented Differentiation Theory, the Mutation Theory, the Error Catastrophy Theory and the Degradation Deficiency Theory with a newly worked out Virus Theory of the cellular aging and the cellular neoplastic transformation. Quantitative in vitro studies of embryonic fibroblast cell systems of two closely related inbred rat strains L.BN and Lewis were undertaken. The data obtained from the experimental analysis of the molecular mechanisms of the cellular aging and the cellular neoplastic transformation demonstrate, that the cellular aging of dividing cells is a Three-Stage-Differentiation Sequence under the control of three different genetic programs. The genetic constitution of the senescent cell regulates the expression of virogenes and oncogenes of the endogenous RNA tumor viruses of the C-type, resulting either in the cellular degeneration of the senescent cell under the control of the virogenes or in the cellular neoplastic transformation regulated by the oncogenes.


Correlation between DNA repair of embryonic fibroblasts and different life span of 3 inbred mouse strains. Primary mouse fibroblast cultures were established from 10 day old embryos of 3 inbred strains with a genetically determined different life expectancy. The capacity for unscheduled DNA synthesis following u.v. irradiation was studied in these cells at various passage levels. The mouse fibroblasts show considerable repair synthesis corresponding to the duration of exposure time. The capacity for induction of unscheduled DNA synthesis was different in the cells of each strain and correlated to the natural life span of the animal; in each case, however, the ability to perform repair synthesis was subjected to an age-associated decline.


[Stimulation of enzymes of red blood cells by pyridoxal-5-phosphate in dependence on age (author's transl)]. In 30 students and 43 patients over 60 years the stimulation of enzymes in erythrocytes by pyridoxal-5-phosphate was tested. GOT in blood and erythrocytes was in the old patients diminished and showed higher values after stimulation.


[Age-dependent DNA-content of brain, testis and ovar in men and rats (author's transl)]. The DNA-content of brain, testis and ovar of Wistar rats (inbred strain) was quantitatively analysed. We observed no significant differences between the several parts of the cerebrum, but between the cerebrum and the cerebellum. Furthermore we demonstrated alterations (with increase and decrease) of the DNA content of all brain parts as well as of testis and ovar during the course of life of this Wistar rat strain. These results and further investigations of the water content as well as of the absolute and of the relative weight of brains of these rats (in comparison to humans) extend the data and our knowledge about these subjects. It is possible to differentiate growth phases (by means of the DNA assays, which correlate with the morphologically well defined growth phases. Finally these results are fundamentals and presuppositions for investigations of the production metabolism of these organs because the DNA content is a better parameter (reference or relation figure, resp.) than the dry or wet weights and the protein or the nitrogen contents.


[Age-dependent enzymatic changes in human cerebral cortex (author's transl)]. Little information about the possible neurochemical-enzymatic changes occuring during aging of human brain is available. We, therefore, investigated the activity of various enzymes of human brains obtained at autopsy and covering a range from 19 to 91 years. Protein kinase, which mediates the information carried by the second messenger cAMP, does not show age-related changes of basal activity. Cyclic AMP-dependent activation of protein kinase remains nearly constant up to 60 years of life, but undergoes a distinct and progressive decline between 60 and 90 years. In corpus striatum no age-related changes of cyclic AMP-dependent protein kinase activity were observed. The activity of carbonic anhydrase demonstrates in both human cortex and corpus striatum an age-dependent decrease which also begins after the 6th decade of life. These neurochemical changes are similar to morphological and physiological changes occuring in the aging brain. They begin after the 60th year of life.


[Adenyl cyclase activity and the stimulation of adenosine 3',5'-cyclic monophosphate in blood vessel walls during ageing and under stress (author's transl)]. It is well-known that the cAMP-adenylate cyclase system is important in mediating the effects of numerous hormones. We investigated the age-dependent behaviour of this system in aortas and femoral arteries of male Wistar rats (at the age of 10 days, 1, 4, 8, 12, and 22 months). It was found that: The basic adenylate cyclase activity had considerably been decreased beyond the first month of life, thereafter it was almost constant. The response of adenylate cyclase to guanylyl-imidodi-phosphate and NaF had essentially been elevated since the 4th month of age. The possibility of stimulating the cAMP generation due to epinephrine and histamine had substantially been increased since the 12th month of age. Besides the ability of adrenaline and histamine to stimulate the formation of cyclic AMP was investigated in broken cell preparation and intact cells of smooth muscle of the aorta and femoral artery of rats which had been subjected to daily intermittend immobilization of 1, 3, and 17 weeks. It was found that this type of stress led to an instability of the blood pressure which was associated with an increase in the sensitivity of adenylate cyclase in the broken cell preparations from the arteries to adrenaline and histamine and with a heightened cyclic AMP response to the two hormones in the intact arterial smooth muscle cells. The sensitivity of cardiac adenylate cyclase for adrenaline remained unchanged.


[Histamine H2-receptor blockade and thrombosis prophylaxis with heparin in tetanus. A case report (author's transl)]. The use of "low-dose" heparin for the purpose of the prophylaxis of thrombosis is not yet general in the treatment of tetanus because this therapy may increase the risk of bleeding gastrointestinal stress-ulcers. The combination of heparin with histamine H2-receptor blockers, however, may be of therapeutic benefit and may reduce this complication as well as thrombosis. A case of tetanus and its therapy is reported.


[Neurostimulation--endorphins. Personal opinion in the guise of an introduction]. Up until 1974 & 1975, Anesthesiologie observed with a certain reserve progress, which was nevertheless spectacular, in the physiological aspects of pain. The fundamental theory of the inhibition of small algophoric fibres by large fibres (NOOR-DENBOS, 1959), the striking theory of spinal control (WALL and MELZACK, 1965) and even the detection of morphine receptors at the same time paradoxically were associated with little enthusiasm here with only a minimum of mention in the journals. Why this lack of enthusiasm? Undoubtedly because new theories remain speculative when viewed from our own standpoint. Since we have been in existence, our methods of general anesthesia have been based upon solid pharmacological concepts but generally paid little attention to physiology which was of relatively little use and even failed to shed any light upon the pharmacology already mentioned. Our attitude was diametrically opposed to that of neurosurgery which by contrast remained within the technical furrow of physiology. The arrival of encephalins forces us to leave behind such attitudes for three reasons: 1--they clearly throw light into the darkness of a whole area of pharmacology; 2--they open up a pharmacological persepective; 3--they raise a number of theoretical and practical questions, which range from their phylogenesis to the hope of one day possessing morphine-like substances without side-effects.


[Current data on pain pathways].

[Current data on pain pathways]. 1--The classical notion attributing the conduction of painful messages to only the A delta and C fibres will be discussed. 2--Spinal cord cells of layers I, V and VIII received painful messages and their physiological properties will be compared. 3--The origin within the cell layers of the spinal cord of the various ascending bundles will be examined. 4--The reticular an thalamic zones (posterior ventra nucleus, mid-line and intralaminar nuclei, GMme posterior group in particular) which receive painful messages have also been studied using activation provoked by pinching, pinprick, the injection of painful substances and stimulation of the dental pulp. The possible role of the different structures thereby brought into action and the limitation of these techniques will be discussed. 5--The existence of metameric controls and controls of central origin active at a spinal, reticular and thalamic level. The absence of inhibitory controls may be responsible for the development of abnormal painful sensations which accompany certain deafferentation procedures. The reestablishment of these controls with the aid of stimulation may result and often results in the improvement of these painful syndromes. 6--Section of the dorsal routes in the chronic animal (rat and cat) may represent an experimental model for deafferentation pain seen in man.


[Pharmacology of morphine and its derivatives (review)]. Morphine, the principal alkaloid of "papaver somniferum" is the reference substance of central analgesics, the parmacodynamic constants of which are: analgesia and the possibility of addiction. Respiratory depression is, for many of them, a grave side-effect. At the present time, no substance in this category is fully satisfactory and all may result in dependence. Equi-analgesic doses of dextromoramide, phenoperidine and Fentanyl are less than those of morphine, whilst those of pethidine and pentazocine are higher. Study of the pharmacokinetics of these various substances indicates no common elements, and it is difficult to consider that the analgesic action is proportional to blood levels. Clinical assessment of the mean duration of action makes it possible to divide morphine derivatives into substances with a very short action (20 to 45 minutes) such as Febtanyl and phenoperidine, and those with a longer action (1 to 4 hours) which includes the majority of the other substances. The analgesic activity of Methoadone lasts for 4 to 6 hours. Morphine antagonists such as Methadone, nalophine, naloxone and naltrexone possess specific problems in terms of their utilization. Pharmacological data concerning theses substances are described.


[Morphine analgesia: neurobiologic data]. Recent Neurobiological (Neurophysiological, Histochemical, Neurochemical and behavioural studies) data have indicated that the analgesic effects of morphine may, at least in part, be explained by two modes of action. A--Morphine has a direct depressive action at a spinal level on the activity of neurones of the grey matter of the dorsal horn which run towards the higher centres of the encephalon. These effects are exerted preferentially on activities induced by the activity of fine non-myelinized fibres (C). These mechanisms are discussed taking into account recent data concerning polypeptides (substance P and encephalins). B--Morphine acts at the level of the brain stem (periaqueductal grey matter, raphian nucleus, etc.) reinforcing the activity of descending bulbo-spinal pathways which block the transmission of painful messages within the cord.


[Opiate receptors and endorphins at the central nervous system level]. Four years ago, sterospecific sites for the bending of opiates were discovered within the brain of animals and the human being. All of the properties of these sites are in conformity with the proposition that they are pharmacological receptors which have long been postulated for these drugs. The binding of morphine or of one of its derivatives to these sites should result in chemical or physical reactions leading to well known pharmacological responses. These reactions following the binding of drugs to the receptors are not yet known, but there is some evidence that cyclical nucleotides play a role. The affinity of a whole series of morphine derivatives, agonists and atagonists, is well correlated with their pharmacological effectiveness. In the presence of sodium salts, antagonists become more strongly bound and agonists less strongly than in the absence of sodium. The evidence is presented. This is explained by an equilibrium between two formations of the receptor: one characteristic of the absence of sodium and one of its presence. Receptors are found in the nervous system of all vertebrates and their distribution has been studied in the human brain. The regions with the highest concentration of receptors are those of the limbic system. A high level exists also in the "substantia gelatinosa" of the spinal cord, which is involved in the passage of painful messages. Study of the function of morphine receptors has led to the isolation, in animal brain, of a number of peptides with morphine properties named endorphines. The first two endorphines isolated were pentapeptides named encephalins. The properties of endorphines from the subject of several lecture in this course.


[Electrophysiological effects of opiates and endogenous peptides on central neurons]. The influence of micro-iontophoretically administered opiates and opiate peptides on the activity of single neurones in the central nervous system will be discussed. It will be shown that in single neurone studies in several different supraspinal regions opiates and opiate peptides have similar actions on individual neurones and both may excite or depress neuronal activity. The excitatory and inhibitory effects of these substances can be reversed by a necrotic antagonist and therefore may be mediated via an action at specific opiate receptors. Studies on spinal neurons indicate that the excitatory action of morphine and enkephalin may be due to an interaction with a cholinergic receptor or at least cholinoceptive neurones. Evidence will also be presented showing that responses of dorsal horn neurones to nowious peripheral stimuli may be selectively reduced by morphine and enkephalin administered into the substantia gelatinosa region or the spinal cord. The relevance of these observations to the pharmacological action of opiates and the possible physiological function of endogenous opiate peptides will be mentioned.


[Transcutaneous electrotherapy in the treatment of chronic post-zoster pain]. Transcutaneous electrotherapy has been employed in 112 patients with chronic pains of various origins. The immediate and long term results are not considered, at whole, satisfactory, especially for length and quality of the obtained improvement. Only patients with chronic post-zoosterian neuralgia had good results, or immediate either at long term, especially in intercostal pains. In the contrary in acute cases of this nevralgia, there were not satisfactory improvements.


[Effects of percutaneous heterosegmental electric stimulation (electro-acupuncture) on the nociceptive flexion reflex in man]. A comparative study of the effects of electro-acupuncture of the HO KU point and of a placebo control point were carried out in man, taking as an objective test of pain the threshold of a pain induced flexion reflex. In the absence of electrical stimulation of the HO KU point, or of the control point, no variation in the painful reflex threshold was seen. This remained stable throughout the experimental session. However, electrical stimulation of the HO KU point resulted in a 115 p. 100 increase in the reflex threshold, whilst that of the control point resulted in an increase of 20 p. 100 in the threshold of the same reflex. These results are discussed on the basis of data in the literature.


[Modulation of experimental dental pain in man with acupuncture and by transcutaneous electric stimulation]. We have studied the effects of electrical acupuncture stimulation and transcutaneous electrical stimulation on the ability of human volunteers to perceive pain. Both psychophysical indices and measures of cerebral evoked responses to painful stimuli have been used to demonstrate that these treatments reduce pain perception. A recently completed study showed that the analgesic effects of transcutaneous electrical stimulation can be partly reversed when 0,4 mg of naloxone is injected. This observation suggests that endogenic morphine-like peptides are released in response to low frequency electrical stimulation of the skin.


[Modes of action of various aspects of analgesia by stimulation]. A study of 12 patients suffering from chronic lumbar pain led to a comparison of the results of treatment with traditional acupuncture and that with transcutaneous electrical stimulation. The degree and quality of pain, as well as the duration of relief after treatment, were analyzed in an attempt to differentiate between the two forms of analgesia. Statistical analysis of the two methods showed no significant difference. Not only did they appear to have the same therapeutic merit, but it would also seem reasonable to suppose that they have a common mechanism at their origin. Intense stimulation of trigger points followed by relief of pain led us to compare them to acupuncture zones both in terms of topography and characteristics. There was a certain analogy of distribution, associated with a similarity of expression which led us to consider that these two techniques were possibly related to the same neurological process.


[Chronic intracerebral electric stimulation for analgesia in man]. This report concerns the current status of the results of intraencephalic implantations carried out up to the present time in man with the aim of relieving certain forms of chronic pain. It places little emphasis upon the neuro-physiological basis of these implantations, which have been studied at length during previous reports. After presenting the overall results of operations carried out in the world up until September 1967, a critical study is undertaken: 1--In relation to the targets (Postero-Lateral Ventral Nucleus, posterior arm of the internal capsule, para-ventricular thalamic grey matter). 2--In relation to etiologies, both analytically (syndrome) by syndrome as well as in terms of synthesis (excess of pain perception-deafferentation). 3--In terms of various factors, such as: --technical requirements; --side ffects; --duration of "post effect" (residual analgesia after stimulation); --parallel action of drugs. Conclusions are drawn concerning future perspectives of this technique.


Differential responsiveness of central noradrenergic and dopaminergic neuron tyrosine hydroxylase to hypophysectomy, ACTH and glucocorticoid administration. Hypophysectomy leads to a small increase in tyrosine hydroxylase activity of all brain areas containing noradrenergic neurons or tuberoinfundibular dopamine neurons, but nigroneostriatal dopamine neurons are not so affected. ACTH or corticosterone treatment inhibited this effect of hypophysectomy in some noradrenergic neurons and in tuberoinfundibular dopamine neurons. These data showing differential responsiveness of tyrosine hydroxylase in different brain areas are compatible with differences in regulation or molecular form of tyrosine hydroxylase in central noradrenergic and dopaminergic neurons. The disparity between increased hypothalamic tyrosine hydroxylase activity and decreased norepinephrine turnover following hypophysectomy may result from a change in the rate-limiting step to the hydroxylation of dopamine.


[Polyol dehydrogenases in mycobacteria (author's transl)]. Contrary to the the tubercle bacilli (H37Ra, BCG), Mycobacterium phlei, grown on Sauton medium, formed the NAD+ dependent dehydrogenases that catalyse the oxidation of ribitol, sorbitol and mannitol. These enzymes were separated by chromatography on DEAE-cellulose and Sephadex G-200. In the present work we have principally studied the ribitol dehydrogenase. All the experiments for induction of the ribitol dehydrogenase in H37Ra or BCG were negative; whereas after the adaptation of M. phlei to ribitol, the specific activity of this enzyme increased in the supernatants more than 100 per cent. The ribitol dehydrogenase of M. phlei reduced NAD+ not only in the presence of ribitol but also (though to a lesser extent) in the presence of erythritol and glycerol. Other properties studied concerning this enzyme and the reaction it catalyses were: pH dependence, equilibrium constant, Km and sensitivity towards the inhibitors of the thiol groups.


Ocular hypotensive efficacy of topical epinephrine in normotensive and hypertensive rabbits: continuous drug delivery vs eyedrops. The ocular hypotensive efficacy of continuously delivered epinephrine is compared to that of pulsed doses provided by eyedrops in both normotensive and hypertensive rabbit eyes. In normotensive eyes, 2 microgram/hr and 4 microgram/hr epinephrine delivered into the tear film continuously for 12 hours reduces intraocular pressure as well as eyedrop pulses of 0.5% epinephrine hydrochloride or 2% epinephrine bitartrate (doses of 500 and 1100 microgram, respectively). Ocular hypertension induced by an intragastric water load in rabbits is significantly inhibited by continuous delivery of epinephrine at the rates of 3 or 6 microgram/hr, or by 2% epinephrine bitartrate (1% free base) eyedrops. Epinephrine delivered continuously at rates of 2-6 microgram/hr for 6 to 12 hours (12-72 microgram total) has hypotensive efficacy equivalent to 15 to 40 times as much epinephrine applied once in eyedrops. Epinephrine bitartrate eyedrops reduce tear film pH well below normal. Continuous delivery of epinephrine bitartrate does not reduce tear film pH below normal levels.


Improved methods for detecting enteric viruses in oysters. New and improved methods for concentrating enteroviruses, reoviruses, and adenoviruses from oysters have been developed and evaluated. Viruses are efficiently adsorbed to homogenized oyster meat by adjusting the homogenate to pH 5.0 and a conductivity of less than or equal to 2,000 mg of NaCl per liter. After low-speed centrifugation, the virus-free supernatant is discarded and the viruses are eluted from the sedimented oyster solids with pH 7.5 glycine-NaCl having a conductivity of 8,000 mg of NaCl per liter. The oyster solids are removed by low-speed centrifugation and filtration, and the viruses in the filtered supernatant are concentrated to a small volume by either ultrafiltration or acid precipitation at pH 4.5. The concentrate is treated with antibiotics and inoculated into cell cultures for virus isolation and quantitation. When these methods were tested with oysters experimentally contaminated with polioviruses, reoviruses, and adenoviruses, recovery efficiencies averaged about 46%. With the exception of virus assay and quantitation, these methods are simple and inexpensive enough to be done in typical shellfish microbiology laboratories.


Comparative effects of ammonia and related compounds on poliovirus. The abilities of ammonia and related compounds to inactivate poliovirus were compared. Compounds virucidal at pH 9.5 had the following order of activities: ethylamine greater than propylamine, dimethylamine, methylamine greater than ammonia greater than 2-methoxyethylamine.


Characteristics of bacteriophages for Micromonospora purpurea. Chemical and physical stabilities of bacteriophages øUW 21 and øUW 51 infecting Micromonospora purpurea ATCC 15835 were examined. Both phages were stable over the pH range of 5 to 8 and to heating at temperatures up to 50 degrees C and especially stable in buffer containing magnesium ion. Exposure to 1 M Ca(NO3)2 inactivated both phages, and phage øUW 51 was also susceptible to 1 M CaCl2, 0.1 M tris(hydroxymethyl)aminomethane, and 0.3% H2O2. Phage plating efficiency was highest on the cultures at logarithmic phase and sometimes much influenced by host growth. Phage øUW 51 has a latent period of 2 h at 34 degrees C and a burst size between 35 and 40. The latent period for phage øUW 21 is about 12 h, and the burst size is smaller than 30.


Inactivation of enteroviruses by ascorbic acid and sodium bisulfite. Poliovirus type 1, coxsackievirus type A9, and echovirus type 7 were inactivated by sodium bisulfite and ascorbic acid. Inactivation rates depended upon concentration, temperature, and pH. RNA infectivity was lost during inactivation; the capsid was also altered by these inactivating agents, as determined by enzyme sensitivity assays and by tests of adsorption to cells. Structural modifications of the virus particles were not identical, suggesting that the mechanism of inactivation by ascorbic acid differs from that of sodium bisulfite.


Development of quantitative methods for the detection of enteroviruses in sewage sludges during activation and following land disposal. The development and evaluation of methods for the quantitative recovery of enteroviruses from sewage sludge are reported. Activated sewage sludge solids were collected by centrifugation, and elution of the solid-associated virus was accomplished by mechanical agitation in glycine buffer at pH 11.0. Eluted viruses were concentrated either onto an aluminum hydroxide floc or by association with a floc which formed de novo upon adjustment of the glycine eluate to pH 3.5. Viruses which remained in the liquid phase after lowering the pH of glycine eluate were concentrated by adsorption to and elution from membrane filters. The method of choice included high pH glycine elution and subsequent low pH concentration; it yielded an efficiency of recovery from activated sludge of 80% for poliovirus type 1, 68% for echovirus type 7, and 75% for coxsackievirus B3. This method was used to study the survival of naturally occurring virus in sludge at a sewage treatment plant and after subsequent land disposal of the solids after aerobic digestion. Reduction of enterovirus titers per gram (dry weight) of solids were modest during sludge activation but increased to a rate of 2 log 10/week after land disposal.


Hydrogen peroxide and superoxide radical formation in anaerobic broth media exposed to atmospheric oxygen. Fourteen different broth media were autoclaved under anaerobic conditions and then exposed to atmospheric oxygen. The hydrogen peroxide and superoxide radical formation as well as the bactericidal effect of the media were studied. The rate of killing of Peptostreptococcus anaerobius VPI 4330-1 was high in media that rapidly autoxidized and accumulated hydrogen peroxide. In actinomyces broth (BBL), 50% of the cells were killed within 2 min, and in Brewer thioglycolate medium (Difco), 50% were killed within 11 min, whereas more than 50% of the cells survived for more than 2 h in Clausen medium (Oxoid), fluid thioglycolate medium (BBL), and thioglycolate medium without dextrose or indicator (Difco). Only media that contained phosphate and glucose had a tendency to accumulate hydrogen peroxide. A solution of phosphate and glucose autoxidized when it had been heated to 120 degrees C for at least 5 min and when the pH of the solution was higher than 6.5. Transitional metal ions catalyzed the autoxidation, but they were not necessary for the reaction to occur. Of the other substances heated in phosphate buffer, only alpha-hydroxycarbonyl compounds autoxidized with accumulation of hydrogen peroxide. Superoxide dismutase decreased the autoxidation rate of most of the broth media. This indicated that superoxide radicals were generated in these media.


Presence and partial characterization of internal acid protease of Aspergillus oryzae. The presence and partial characterization of the internal acid protease (EC 2.4.23.6) of Aspergillus oryzae has been investigated. Although the majority of the acid protease is external and present in the culture filtrate, a significant amount of the active enzyme is firmly bound to the cells; it is not released by repeated extraction of cells with 0.9% sodium chloride but is liberated into the soluble fraction during disruption of cells. The internal acid protease, as well as the external one, was separated into two major molecular forms (F1 and F2) with molecular weights of 60,000 and 42,000, respectively, by chromatography on Sephadex G-100 and on CM-Sephadex C-50. The partially purified internal enzymes had the same catalytic and immunological properties, as did the external enzyme.


Prevalence of fungi during Skylab missions. Samples for mycological analysis were collected from surfaces in the Skylab spacecraft before launch and during flight for each manned mission. Fungal contamination levels were low during the first two flights; however, the species recovered were different for each mission. On the third mission, widespread contamination of the Skylab spacecraft with Aspergillus and Pencillium spp. was detected. This contamination was traced to several contaminated space suit undergarments.


Phthalate and 4-hydroxyphthalate metabolism in Pseudomonas testosteroni: purification and properties of 4,5-dihydroxyphthalate decarboxylase. Phthalate is degraded through 4,5-dihydroxyphthalate and protocatechuate in Pseudomonas testosteroni NH1000. The ezyme 4,5-dihydroxyphthalate decarboxylase, catalyzing the conversion of 4,5-dihydroxyphthalate to protocatechuate and carbon dioxide, was purified approximately 130-fold from phthalate-induced cells of a protocatechuate 4,5-dioxygenase-deficient mutant of P. testosteroni. The most purified preparation showed a single protein band on sodium dodecyl sulfate-acrylamide disc gel electrophoresis with a molecular weight of 38,000. The apparent molecular weight of the native enzyme determined by Sephadex G-200 column chromatography was 150,000. Among the substrate analogs tested, only 4-hydroxyphthalate served as a substrate, which was decarboxylated to form m-hydroxybenzoate. The apparent Km values for 4,5-dihydroxyphthalate and 4-hydroxyphthalate were estimated to be 10.5 micrometer and 1.25 mM, respectively, and the Vmax for the former was 10 times larger than that for the latter. Whereas the wild-type strain could utilize 4-hydroxyphthalate as a sole source of carbon, none of the following could grow with the compound: 4,5-dihydroxyphthalate decarboxylase-deficient, m-hydroxybenzoate-nondegradable, and protocatechuate 4,5-dioxygenase-deficient mutants. Since one-step revertants of these mutants could utilize 4-hydroxyphthalate, the compound appears to be metabolized through m-hydroxybenzoate and protocatechuate in P. testosteroni NH1000.


Tissue sterility in uneviscerated carcasses. Sheep muscle tissue removed aseptically from control carcasses, from uneviscerated carcasses held at 20 degrees C for 24 h, and from carcasses of sheep subjected to stress before slaughter was examined for the presence of bacteria. All samples from a total of 68 carcasses were sterile. Whole-body autoradiography of mouse carcasses showed that 14C-labeled fixed bacteria injected after death remained in the lumen of the intestine. Live bacteria did not penetrate the mucosal surface until the tissue structure had been disrupted by proteolytic enzymes. Bacteria were unable to penetrate sections of intestine longitudinally until considerable structural breakdown had occurred, indicating that blood and lymph vessels do not normally offer a pathway for microbial invasion from the intestine. Clostridia, which have been reported to be responsible for deep spoilage of meat, reached maximum numbers 24 to 28 h after death in the intestines of guinea pig carcasses stored at 20 degrees C, but did not invade carcass tissues until the stomach ruptured as a result of proteolysis between 2 and 3 days after death.


Storage quality of dark, firm, dry meat. Dark, firm, dry meat contains little or no glucose. Therefore, spoilage bacteria growing on such meat immediately attack amino acids so that spoilage odors and ammonia are detectable at comparatively low cell densities (about 10(6)/cm2).


Establishment of a heat inactivation curve for Clostridium botulinum 62A toxin in beef broth. A procedure is described for establishing a heat inactivation curve for the toxin of Clostridium botulinum 62A in beef broth. The effect of toxin titer, pH, and the type of acid employed for pH adjustment on the heat stability of the toxin is described.


Membrane bound pyrophosphatases in Entamoeba histolytica. Entamoeba histolytica homogenates are capable of hydrolyzing a range of inorganic and organic pyrophosphates. Two separate activities are present: an inorganic pyrophosphatase hydrolyzing inorganic pyrophosphate and linear tripolyphosphate, and a nucleoside diphosphatase hydrolyzing thiamine pyrophosphate and nucleoside diphosphates (ADP, GDP and UDP). The inorganic pyrophosphatase has an acid pH optimum, a relatively high KM (congurent to 1 micrometer) and is markedly heat stable and lacks a metal requirement. The nucleoside diphosphatase also has an acid pH optimum but displays a much higher affinity for substrate (KM congurent to 50 micrometer), is unstable to heating and is activated by Ca ions. Both pyrophosphatases distinct from the acid phosphatase activity which is also present. All three hydrolases are sedimentable and latent suggesting their association with membrane bounded organelles. No soluble inorganic pyrophosphatase activity could be demonstrated.


Fine structural localization of alkaloid synthesis in endoplasmic reticulum of submerged Claviceps purpurea. Acetyl coenzyme A (CoA) carboxylase (EC 6.4.1.2), an enzyme catalyzing the synthesis of malonyl-CoA, was cytochemically localized in endoplasmic reticulum (ER) of sclerotia-like cells of submerged Claviceps purpurea Tul. producing clavine alkaloids. The enzymic activity was structurally bound in unit membranes of ER strands which, later on, evolved into vacuoles containing lipoprotein material. The reaction product was absent from ER in nonvacuolized filamentous hyphae and ovoid asexual spores containing numerous lipid globules; it was also absent from ER in the mycelium of submerged C. purpurea strain producing no alkaloids. In view of our previous morphogenetic observations and the available biochemical evidence, the observed localization of acetyl-CoA carboxylase was assumed not to coincide with fatty acid biosynthesis but to represent sites of alkaloid synthesis.


Evidence for an incomplete reductive carboxylic acid cycle in Methanobacterium thermoautotrophicum. The involvement of reactions of the tricarboxylic acid cycle in autotrophic CO2 fixation in Methanobacterium thermoautotrophicum was investigated. The incorporation of succinate into glutamate (= alpha-ketoglutarate), aspartate (= oxaloacetate) and alanine (= pyruvate) was studied. The organism was grown on H2 plus CO2 at pH 6.5 in the presence of 1 mM [U-14C-]succinate. Significant amounts of the dicarboxylic acid were incorporated into cellular material under these conditions. Alanine, aspartate, and glutamate were isolated and their specific radioactivities were determined. Only glutamate was found to be labelled. Degradation of glutamate revealed that C-1 of glutamate was derived from CO2 and C-2--C-5 from succinate indicating that in M. thermoautotrophicum alpha-ketoglutarate is synthesized via reductive carboxylation of succinyl CoA. The finding that succinate was not incorporated into alanine and aspartate excludes that oxaloacetate and pyruvate are synthesized from alpha-ketoglutarate via isocitrate or citrate. This is taken as evidence that a complete reductive carboxylic acid cycle is not involved here in autotrophic CO2 fixation.


Cellulose degradation and cellulase formation by Phialophora malorum. The formation of cellulases and beta-glucosidase and their location in the fungus Phialophora malorum was studied on some different carbon sources. The cellulases were found to be partly cell-free and partly cell-bound during growth on cellulose and carboxymethyl-cellulose. Glucose and cellobiose repressed the cellulase formation but a low carboxymethylcellulase activity was measurable on the glucose-grown mycelium. The unicellular stage did not appear to grow on carboxymethyl-cellulose or cellulose, but mycelium was formed on these carbon sources.


Incorporation of labeled small molecules into rubratoxin. A sterile glucose-mineral salts broth was inoculated with conidia of Penicillium rubrum P-13 and P-3290. Radiolabeled compounds were added to some cultures, these being incubated quiescently at 28 degrees C for 14 days. Other stationary cultures were grown for 21 days, received labeled compounds, and were then grown for 5 more days. The remaining cultures were inoculated with 72-h-old mycelial pellets, received labeled materials and were incubated with shaking for 60 h. Rubratoxin was resolved by thin-layer chromatography. Labeled [1(14)C]acetate, [1,5(14)C]citrate, [2(14)C]malonate, [1(14)C]glucose, [U14C]glucose or [1(14)C]hexanoate were incorporated into rubratoxins A and B by P. rubrum 3290 and into rubratoxin B by P. rubrum 13. Incorporation of [1(14)C]acetate and [2(14)C]malonate increased when exogenous unlabeled acetate, malonate, pyruvate, or phosphoenol-pyruvate was added. Acetate incorporation was influenced by cultural conditions, attaining maximum amounts in quiescent cultures which received labeled acetate after 21 days of incubation. Acetate incorporation in shake cultures was enhanced by reduced nicotinamide adenine dinucleotide phosphate (NADPH) and by unlabeled exogenous citrate.


Utilization of D-amino acids by dadR mutants of Salmonella typhimurium. Utilization of D-amino acids being substrates of D-amino acid dehydrogenase of Salmonella typhimurium was examined. The experiments were done with wild type strains and the mutants dadA missing the enzyme activity and dadR in which its synthesis is released from catabolite repression. Growth on D-tryptophan, D-histidine and D-methionine used as precursors of the L-amino acids was faster when the respective auxotrophs carried dadR mutations. The dadR mutants grew faster when D-or L-alanine was present as a sole source of nitrogen. Experiments with D-amino acid dehydrogenase in vitro provided evidence that D-tryptophan is its substrate with a very low affinity to the dehydrogenase.


Relation of plasma prolactin to clinical response in schizophrenic patients. It has been suggested that, if dopamine antagonism is a necessary condition for the antischizophrenic action of neuroleptics, the prolactin response, as an index of dopamine blockade, would correlate with clinical response. Morning prolactin and clinical symptomatology were measured in 15 schizophrenic patients before neuroleptic therapy, and after three and six weeks of high-dose butaperazine or loxapine treatment. Prolactin levels were transiently elevated during the unmedicated admission period, probably reflecting a normal stress response. Prolactin increased in all patients during neuroleptic therapy. There was, however, no correlation between magnitude of prolactin changes and clinical response, probably because the prolactin response achieved a maximum at relatively low doses of neuroleptics.


Monozygotic twins with presumed metachromatic leukodystrophy. Activity of arylsulfatase A in serum of patients and family. Arylsulfatase A (ASA) activity in urine and serum was assayed on two 21-month-old monozygotic twins with presumed metachromatic leukodystrophy (MLD), their parents, and kin. The patients showed a marked reduction in ASA activity in both urine and serum. The twins' parents and 11 kin, a total of 13 persons, were examined for ASA activity in serum, but it was not possible to delineate heterozygous carriers of MLD by the present study. The assay of ASA activity in serum promises to be useful for diagnosis of MLD.


Behavior therapy and the treatment of flight phobia. Research since 1960 was reviewed to compare the relative efficacy of behavior therapy and other therapy modalities in the treatment of flight phobias in trained military aircrew, commercial flight crews, and civilian air travellers. Results indicated that treatment programs involving behavior therapy techniques appeared to have a higher success rate than treatment programs not involving these techniques, especially in cases of focal fear with acute onset under stressful conditions. Other conditions for optimal outcome are outlined and hypotheses for a more programmatic research effort discussed.


Purification and properties of gamma-glutamylcyclotransferase from human erythrocytes. 1. GAMMA-Glutamylcyclotransferase was purified 10000-fold from human erythrocytes. 2. The purification steps involved fractionation with (NH4)(2)SO(4) and chromatography on Sephadex G-75, DEAE-cellulose and hydroxyapatite. The purified enzyme was found to be homogeneous on density-gradient polyacrylamide-gel electrophoresis. 3. The maximum reaction rate was observed at pH9.0 and the apparent Km value for gamma-glutamyl-L-alanine was 2.2mM. 4. The molecular weight (25250) of the purified enzyme agreed well with the value (25500) in fresh haemolysates, indicating no apparent structural modification of the enzyme during purification. However, rapid processing of the blood through the initial (NH4)(2)SO(4) and Sephadex-chromatography steps was required to prevent formation of a high-molecular-weight aggregate with substantially lower specific activity. 5. gamma-Glutamylcyclotransferase catalyses the formation of 5-oxoproline from gamma-glutamyl dipeptides. The role of this enzyme in erythrocytes is of particular interest, because gamma-glutamyl-L-cysteine serves as a substrate for both gamma-glutamylcyclotransferase and glutathione synthetase. Thus the cyclotransferase could modulate glutathione synthesis.


Purification of rat intestinal maltase/glucoamylase and its anomalous dissociation either by heat or by low pH. Maltase-glucoamylase, a microvillous membrane ectoenzyme, was solubilized from rat intestinal mucosa by digestion with papain and subsequently purified to homogeneity with an overall yield of 10--20%. An antibody to the purified enzyme formed a single precipitin line in immunodiffusion experiments with an intestinal homogenate. The enzyme was shown to be an acidic glycoprotein (20% sugar by weight) which contained low amounts of cysteine and no sialic acid. At pH3--6, maltase activity was slowly lost, but the enzyme was re-activated by re-adjustment of the pH to neutrality. However, in the presence of sodium dodecyl sulphate, acid pH values inactivated maltase irreversibly, and at the same time converted the enzyme (mol.wt. 500000 approx.) into five new species with apparent molecular weights ranging from 134000 to 480000 as judged by polyacrylamide-gel electrophoresis. The same five fragments were also formed by boiling the enzyme for brief periods in the presence of sodium dodecyl sulphate or urea either with or without reducing agents. The dissociated species were stable on re-electrophoresis, and amino acid analysis showed them to be very similar to each other and to the original enzyme. The bands migrated anomalously on polyacrylamide gels of different concentration, thereby preventing the assignment of precise molecular weights. It is possible that the five species may represent stable aggregates of a common monomer of the enzyme.


The selective retardation of NADP+-dependent dehydrogenases by immobilized procion red HE-3B. The capacities of Procion Red HE-3B and Cibacron Blue F3G-A immobilized to Sepharose CL-4B and Matrex 201R for NAD+-, NADP+- and NAD(P)+-dependent dehydrogenases were measured. Procion Red HE-3B columns retarded NADP+-dependent dehydrogenases more effectively than NAD+-dependent dehydrogenases, whilst immobilized Cibacron Blue F3G-A retarded NAD+-dependent dehydrogenases more effectively than NADP+-dependent dehydrogenases. The capacity of procion Red HE-3B-Sepharose CL-4B for five dehydrogenases was highest in the region of 70nmol of immobilized ligand/ml of settled gel. The effects of using poly(ethyleneimine) as a spacer for both porous and pellicular supports were also examined. Four NADP+-dependent dehydrogenases were purified from yeast extract by using Procion Red HE-3B-Sepharose CL-4B. Two NAD+-dependent dehydrogenases were purified from the same source using Cibacron Blue F3G-A-Sepharose CL-4B. These results are discussed in relation to the use of immobilized Procion Red HE-3B to purify dehydrogenases. This immobilized dye's chromatograhic behaviour is compared with that of immobilized nucleotides. The most important feature of immobilized tirazine dyes seems to be their high operational capacities when compared with group-specific nucleotide adsorbents.


Malate dehydrogenase of the cytosol. Ionizations of the enzyme-reduced-coenzyme complex and a comparison with lactate dehydrogenase. 1. The pH-dependencies of the binding of NADH and reduced nicotinamide--benzimidazole dinucleotide to pig heart cytoplasmic malate dehydrogenase and lactate dehydrogenase are reported. 2. Two ionizing groups were observed in the binding of both reduced coenzymes to lactate dehydrogenase. One group, with pKa in the range 6.3--6.7, is the active-site histidine residue and its deprotonation weakens binding of reduced coenzyme 3-fold. Binding of both coenzymes is decreased to zero when a second group, of pKa 8.9, deprotonates. This group is not cysteine-165.3. Only one ionization is required to characterize the binding of the two reduced coenzymes to malate dehydrogenase. The group involved appears to be the active-site histidine residue, since its ethoxycarbonylation inhibits the enzyme and abolishes binding of reduced coenzyme. Binding of either reduced coenzyme increases the pKa of the group from 6.4 to 7.4, and deprotonation of the group is accompanied by a 10-fold weakening of coenzyme binding. 4. Two reactive histidine residues were detected per malate dehydrogenase dimer. 5. A mechanism which emphasizes the homology between the two enzymes is presented.


Identification of histidine-122alpha in human haemoglobin as one of the unknown alkaline Bohr groups by hydrogen--tritium exchange. Human carbonmonoxy- and deoxy-haemoglobins were incubated at 37 degrees C in 3H2O at various pH values to measure the pH-dependent hydrogen--tritium exchange at the C-2 position of the imidazole ring of histidine-122alpha. To obtain the pseudo-first-order rate constants for the exchange, k, the two peptides containing histidine-122alpha were isolated and the amounts of tritium incorporated were determined. The rate constants gave pK values for the histidine of 6.1 in carbonmonoxyhaemoglobin and 6.6 in deoxyhaemoglobin, showing that it contributes about 20% to the total alkaline Bohr effect and about 10% at pH7.4.