The effects of the ultraviolet-protecting plasmids pKM101 and R205 on DNA polymerase I activity in Escherichia coli K-12. The mutagenesis- and repair-enhancing plasmids pKM101 and R205 were introduced into a series of Esherichia coli K-12 polA mutants including two temperature-sensitive mutants. Polymerase levels in extracts of these strains were assayed using an activated DNA template. In none of the cases did the presence of the plasmid in the strains change either the initial rate of incorporation of [3H]thymidine triphosphate into acid-soluble material or the subsequent degradation of the template at longer reaction times. Neither did the presence of the plasmids affect the proportion of N-ethylmaleimide-sensitive polymerase activity detected. Previous studies have reported increased polymerase I-like activity of polA mutants of Salmonella typhimurium and Pseudomonas aeruginosa upon introduction of mutagenesis- and repair-enhancing plasmids. Our experiments indicate that, at least, such an increase in polymerase-I-like activity is not an obligatory phenotype associated with these plasmids.


[Heat denaturation of paramyosin].

[Heat denaturation of paramyosin]. Heat denaturation of paramyosin and its two proteolytic fragments has been studied by scanning microcalorimetry. It has been shown that paramyosin denaturation proceeds in several stages, each stage corresponding to the melting of a cooperative block in the macromolecule. The junctions between the blocks are the most strongly affected by proteolytic fragments. Enthalpies of paramyosin and its fragment denaturation and their temperature dependencies have been measured. It has been shown that the mean value of the specific enthalpy extrapolated to 110 degrees is noticeably lower than for globular proteins and is close to the values obtained earlier for L-meromyosin and tropomyosin. A conclusion has been drawn that the lower, in comparison with globular proteins, value of the denaturation enthalpy of alpha-superhelical structure is of a general character.


The mutagenicities of safrole, estragole, eugenol, trans-anethole, and some of their known or possible metabolites for Salmonella typhimurium mutants. Safrole, estragole, anethole, and eugenol and some of their known or possible metabolites were tested for mutagenic activity for S. typhimurium TA1535, TA100, and TA98. Highly purified 1'-hydroxyestragole and 1'-hydroxysafrole were mutagenic (approximately 15 and 10 revertants/micromole, respectively) for strain TA100 in the absence of fortified liver microsomes; trans-anethole and estragole appeared to have very weak activity. 3'-Hydroxyanethole was too toxic for an adequate test. Supplementation with NADPH-fortified rat-liver microsomes and cytosol converted 3'-hydroxyanethole to a mutagen(s) and increased the mutagenic activities for strain TA100 of 1'-hydroxyestragole, 1'-hydroxysafrole, estragole, and anethole. No mutagenicity was detected for safrole or eugenol with or without added NADPH-fortified liver preparations. The electrophilic 2',3'-oxides of safrole, 1'-hydroxysafrole, 1'-acetoxysafrole, 1'-oxosafrole, estragole, 1'-hydroxyestragole, and eugenol showed dose-dependent mutagenic activities for strain TA1535 in the absence of fortified liver microsomes. These mutagenic activities ranged from about 330 revertants/micromole for 1'-oxosafrole-2',3'-oxide to about 7000 revertants/micromole for safrole-2',3'-oxide. The arylalkenes, their hydroxylated derivatives, or their epoxides did not show mutagenic activity for strain TA98, except for 1'-oxosafrole-2',3'-oxide, which had weak activity. Since the arylalkenes are hydroxylated and/or epoxidized by hepatic microsomes, hydroxy and epoxide derivatives appear to be proximate and ultimate mutagenic metabolites, respectively, of the arylalkenes.


[Quantitative studies of interaction of polyuridylic acid with 30S subunits of ribosomes of Escherichia coli]. Polyuridilic acid of average molecular weight 18 000 binds to the 30S subunits with stoichiometry 1 : 1 but two kinds of 30S.poly(U) complexes with different stability are formed. The main reason for such heterogeneity was found to be due to the presence or absence of ribosomal protein Sl in 30S subunits. In its presence the association constant of 30S.poly(U) complex is equal 2.7.10(8) M-1, and in the opposite case it is much less 1.5.10(6) M-1. In the same conditions (20 mM MgCl2, 200 mM HN4Cl, 0 degrees) the association constant of binary complex Sl.poly(U) is equal 5.10(7) M-1.


Reduction in mutagenicity of cigarette smoke condensate by added sugars. The effects of adding sugars to high- and low-tar cigarettes on the mutagenicity of their smoke condensates were studied using Salmonella typhimurium TA100 and TA98 with and without metabolic activation. The sugars tested were glucose, fructose, galactose, sorbitol, sucrose and lactose. The lowest mutagenicities observed with these sugars per mg of smoke condensate assayed on TA98 with metabolic activation were 37% (high-tar cigarettes) and 22% (low-tar cigaretts) of that of smoke condensate from untreated cigarettes. Addition of sugars increased the total amounts of smoke condensates, but the mutagenicities of the total condensates were also decreased by all the sugars, the lowest values being 35% (high-tar cigarettes) and 36% (low-tar cigarettes) of that of smoke condensates from cigarettes without added sugar. On assay with TA100 with metabolic activation, decreases in both specific and total mutangenicities of condensates of high-tar cigarettes were observed with all the sugars tested except galactose and sucrose. Treatment with glucose, fructose or sorbitol decreased the specific mutagenicity of condensates of low-tar cigarettes and glucose and fructose reduced also their total mutagenicity. The effects of added sugars were more marked when assayed on TA98 than on TA100 and of the sugars tested fructose and sorbitol had the greatest effects. Addition of sugars had no effect of the mutagenicity of cigarette-smoke condensate without metabolic activation.


The mutagenic potential of unexcised pyrimidine dimers in Saccharomyces cerevisiae, rad1-1: evidence from photoreactivation and pedigree analysis. Photoreactivation and pedigree analysis have been combined to show that unexcised pyrimidine dimers in the DNA  of rad1-1 yeast can initiate mutagenesis after passing through several DNA replications. Monomerisation of dimers immediately before the second replication to follow UV has no effect on mutants appearing after the first post-UV cell division but reduces second-generation mutants to one third of their frequency in the dark and has a similar through slightly less marked effect on mutants appearing in the third or subsequent generations. The bearing of these findings on the mechanism of UV mutagenesis is dicussed.


Mitochondrial mutagenesis in Saccharomyces cerevisiae. I. Ultraviolet radiation. UV efficiently induces mutations in mitDNA , conferring resistance to erythromycin. Mitochondrial chloramphenicol-resistant mutants are probably also induced by UV, but almost 90% of mutants with such phenotype are non-mitochondrial; therefore it is possible to estimate accurately the frequences of the induced presumptive mitochondrial capr mutations.


Mutagenicity of 43 structurally related heterocyclic compounds and its relationship to their carcinogenicity. 43 heteropolycyclic compounds belonging to a homologous series were investigated for mutagenicity. The results are compared with carcinogenicity data obtained with the same batches of compounds under conditions identical for all of them. Mutagenicity was tested in the Ames test with Salmonella typhimurium strains TA1535, TA1537 and TA100 in the presence and absence of liver 10 000 g supernatant from rats treated with Aroclor 1254. Carcinogenicity was tested by injection of the compounds into subcutaneous tissue of XVIInc/Z mice. 18 test compounds showed carcinogenic activity, some strongly, others only weakly. Of these, 17 were detected as mutagens: one weak carcinogen did not revert the Salmonella strains. No quantitative correlation was observed between the extents of the mutagenic and the carcinogenic effects. Of the 25 substances that did not produce tumours, 13 showed mutagenicity (12 in the presence, 2 in the absence, of the liver homogenate). The mutagenic effects of these compounds were quantitatively similar to those of the compounds that produced tumours. The most sensitive strain of Salmonella typhimurium was TA100. It detected all 30 mutagens. TA98 was mutated by 25 compounds, TA1537 by 16 compounds. No mutagenic effects were seen with TA1535. Possible reasons for the high percentage of apparently "false positives" in the Ames test and the lack of a quantitative correlation between the potency of the mutagenic and carcinogenic effects are discussed. It is suggested that the complexity of the metabolism of these heterocyclic compounds may lead to critical differences in metabolism in mouse subcutaneous tissue in vivo and in liver homogenates from rats treated with Aroclor. Therefore the present study will be extended to life-long oral and intrahepatic carcinogenicity tests leading to a higher proportion of metabolism in the liver.


delta 1-Tetrahydrocannabinol and 1 alpha, 2 alpha-epoxyhexahydrocannabinol: mutagenicity investigation in the Ames test. 1,2-Epoxyhexahydrocannabinol is a metabolite of delta 1-tetrahydrocannabinol. Because many epoxides are mutagens, we investigated 1,2-epoxyhexahydrocannabinol as well as delta 1-tetrahydrocannabinol for mutagenicity with Salmonella typhimurium TA1535, TA1537, TA98 and TA100 in the presence and in the absence of S9 mix from liver homogenate of rats treated with Aroclor 1254. Additionally, an epoxide hydratase inhibitor was used in some experiments. Whereas several other epoxides and further positive controls, not requiring activation or activated under the same conditions, respectively, showed strong mutagenicity, no indications of a mutagenic hazard by 1,2-epoxyhexahydrocannabinol or by delta 1-tetrahydrocannabinol were found.


Mutagenicity of phenanthrene and phenanthrene K-region derivatives. Phenanthrene and 9 K-region derivatives, most of them potential metabolites of phenanthrene, were tested for mutagenicity by the reversion of histidine-dependent Salmonella typhimurium TA1535, TA1537, TA1538, TA98 and TA100 and the rec assay with Bacillus subtilis H17 and M45. The strongest mutagenic effects in the reversion assay were observed with phenanthrene 9,10-oxide, 9-hydroxyphenanthrene and N-benzyl-phenanthrene-9,10-imine. Interestingly, the mutagenic potency of the arene imine was similar to that of the corresponding arene oxide. This is the first report on the mutagenicity of arene imine. The mutagenic effects of all these phenanthrene derivatives were much weaker than that of the positive control benzo[a]pyrene 4,5-oxide. Even weaker mutagenicty was found with cis-9,10-dihydroxy-9,10-dihydrophenanthrene and with trans-9,10-dihydroxy-9-10-dihydrophenanthrene. The other derivatives were inactive in this test. However, 9-10-dihydroxyphenanthrene and 9,10-phenanthrenequinone were more toxic to the rec- B. subtilis M45 strain than to the rec+ H17 strain. This was also true for phenanthrene 9,10-oxide and 9-hydroxyphenanthrene, but not with the other test compounds that reverted (9,10-dihydroxy-9,10-dihydrophenanthrenes; N-benzyl-phenanthrene 9,10-imine; benzo[a]pyrene 4,5-oxide) or did not revert (phenanthrene, 9,10-bis-(p-chlorophenyl)-phenanthrene 9,10-oxide, 9-10-diacetoxyphenanthrene) the Salmonella tester strains. Although the K region is a main site of metabolism and although all potential K-region metabolites were mutagenic, phenanthrene did not show a mutagenic effect in the presence of mouse-liver microsomes and an NADPH-generating system under standard conditions. However, uhen epoxide hydratase was inhibited, phenanthrene was activated to a mutagen that reverted his- S. typhimurium. This shows that demonstration of the mutagenic activity of metabolites together with the knowledge that a major metabolic route proceeds via these metabolites dose not automatically imply a mutagenic hazard of the mother compound, because the metabolites in question may not accumulate in sufficient quantities and therefore the presence and relative activities of enzymes that control the mutagenically active metabolites are crucial. N-Benzyl-phenanthrene 9.10-imine was mutagenic for the episome-containing S. typhimurium TA98 and TA100 but not for the precursor strains TA1538 and TA1535. This arene imine would therefore be useful as a positive control during routine testing to monitor in the former strains the presence of the episome which is rather easily lost.


Antioxidants reduce the mutagenic effect of malonaldehyde and beta-propiolactone. Part IX. Antioxidants and cancer. Increasing concentrations of malonaldehyde and beta-propiolactone were increasingly mutagenic with 7 mutants of Salmonella typhimurium, 5 of which mutated bya frameshift mechanism and 2 of which mutated through base-pair substitution. The antioxidants vitamin C, vitamin E, selenium and butylated hydroxytoluene (BHT) at 3 logarithmic concentrations markedly reduced mutagenesis in those strains which mutated by frameshift mechanism.


Detection of the mutagenic activity of lead chromate using a battery of microbial tests. The potential mutagenicity of the carcinogen lead chromate was tested by the following battery of microbial tests: the Escherichia coli PolA+/PolA- survival test; the Salmonella/microsome His+ reversion assay; the E. coli Trp+ reversion test as a plate assay; the E. coli Gal+ forward mutation test; and the Saccharomyces cerevisiae assay for mitotic recombination. Lead chromate is mutagenic in Salmonella and in Saccharomyces and is thus identified as a microbial mutagen by this battery. Metabolic activation by rat liver homogenate (S9) is not required for the mutagenic activity of lead chromate. The most statistically significant, positive result is found with a supplementary assay, the E. coli fluctuation test. To determine whether the lead ion and/or the chromate ion were responsible for the mutagenicity observed, lead chloride and chromium trioxide (chromic acid) were also tested. In E. coli fluctuation test, the ranges of maximal mutagenicity for chromium trioxide and lead chromate overlap at the concentration 10(-5)M, whereas lead chloride shows no mutagenicity and little lethality at concentrations up to 10(-3)M. Thus, it appears that the chromate ion is responsible for the mutagenicity of lead chromate.


Mutagenic action of a series of epoxides. The mutagenicity of a series of 13 epoxide compounds was studied using a bacterial plate assay system. The histidine-dependent tester strains TA98 (for frameshift mutagens) and TA100 (for base-pair substitution mutagens) of Salmonella typhimurium were used. Mutagenicity was evaluated both with and without the additon of rat liver microsomal extract. Dieldrin, diglycidyl ether of bis phenol A and 3 of its homologues were not mutagenic. Allyl glycidyl ether, n-butyl glycidyl ether, vinly cyclohexene diepoxide, glycidol, glycidal-dehyde, diglycidyl ether, diepoxybutane and diglycidyl ether of substituted glycerine were mutagenic in the TA100 strain, causing reversion of the bacteria to histidine independence. Dose-reponse curves of the mutagenicity of the latter 4 compounds were obtained. On a molar basis, glycidaldehyde was about 20-50 times more potent in producing mutation that were the other 3 epoxides in the dose-response test. In general, the mutagenicity of the epoxides was not enhanced or diminished by the addition of microsomal extract.


The mutagenicity of halogenated alkanols and their phosphoric acid esters for Salmonella typhimurium. 9 halogenated alkanols, 9 corresponding tris (haloalkyl)phosphates, and 2 bis-(2,3-dibromopropyl)phosphate salts were evaluated for mutagenicity against Salmonella typhimurium TA98, TA100, TA1535, TA1537 and TA1538, with and without rat liver in vitro metabolic activation system (S9 mix). Most of the test samples showed mutagenic activity in the strains TA100 and TA1535, but not in the strains TA98, TA1537 and TA1538. In general, the mutagenic activities of the phosphates obtained with S9 mix were greater than the activities obtained without S9 mix. Among the phosphates, several structure--activity relationships were found; i.e., (i) the bromoalkyl derivatives were more mutagenic than the corresponding chloroalkyl derivatives, (ii) the beta-haloethyl derivatives were more mutagenic than the gamma-halopropyl derivatives, (iii) the phosphates having adjacent beta and gamma halogen atoms in the alkyl moiety, e.g., tris-(2,3-dibromopropyl)phosphate, were particularly potent mutagens, (iv) the branched carbon chain reduced the mutagenic activities in spite of the presence of beta-halogen atoms, e.g., tris(1-bromomethyl-2-bromoethyl)phosphate. However, such relations did not necessarily apply to the halogenated alkanols. It is concluded that the metabolic activation pathway via haloalkanols to mutagens must not be in common with all tris-BP-like phosphates.


Mutagenicity test of dyes used in cosmetics with the Salmonella/mammalian-microsome test. 37 dyes including 3 anthraquinone, 22 azo; 5 xanthene, 5 fluorandiol, and 2 thioindigo dyes, were tested for mutagenic potential with the Salmonella/mammalian-microsome test. Two frame-shift histidine mutants (TA1537 and TA98) and two base-pair substituted histidine mutants (TA1535 and TA100) of Salmonella typhimurium were employed. Both the spot test and the plate-incorporation assay indicated that one azo dye, D&C Orange No. 17, was mutagenic with three of the bacterial test strains. The mutagenic response of D&C Orange No. 17 was depressed by the addition of the microsomal fractions from rat livers. Of the chemicals used to synthesize D&C Orange No; 17 was depressed by the addition of the microsomal fractions from rat livers. Of the chemicals used to synthesize D&C Orange No. 17, beta-naphthol was not mutagenic but 2,4-dinitroaniline was mutagenic to the same Salmonella strains as D&C Orange No. 17 . Dimethyl sulfoxide extracts of lipsticks of similar formula but without D&C Orange No. 17 were tested in the plate incorporation assay. Only those containing D&C Orange No. 17 were mutagenic and the dye was mutagenic at concentrations consumed in normal daily use.


Mutagenicity of cyclic nitrosamines in Salmonella typhimurium: effect of ring size. Mutagenicity of cyclic nitrosamines with varying carcinogenic potentials was assayed in the Salmonella histidine-reversion system. Mutagenicity in the pour-plate assay was compared with that in the liquid pre-incubation test. The smaller ring compounds (nitrosoazetidine, nitrosopyrrolidine, and nitrosopiperidine) exhibited a similar effect in both assays. The large ring compounds (nitrosohexamethyleneimine, nitrosoheptamethyleneimine, nitrosooctamethyleneimine, and nitrosododecamethyleneimine) were more effective in the liquid pre-incubation test. Our results suggest a reasonable relationship between their mutagenic and carcinogenic activities.


In vitro mutagenicity and cell transformation screening of phenylglycidyl ether. Phenylglycidyl ether (1,2-epoxy-3-phenoxy propane) (PGE) was tested for genetic activity in bacterial and mammalian tests. It was active in the Salmonella/microsome mutagenicity test. Concentration-dependent mutagenicity was demonstrated in S. typhimurium strains TA1535 and TA100 with and without rat S9, but not in strains TA98, TA1537, or TA1538. These results suggest PGE, is a direct-acting mutagen causing base substitutions. Phenylglycidyl ether did not induce 6-thioguanine-resistant mutants of Chinese hamster ovary cells, with or without rat S9, and with or without serum in the medium. Dose-dependent enhancement of SA7 virus transformation of primary hamster embryo cells was observed at concentrations of 1.6 microgram/ml and higher. In addition, this compound was able to chemically transform secondary hamster embryo cells at concentrations of 6.2 micrograms/ml and higher. At a dose of 2500 mg/kg p.o., PGE was active in the host-mediated assay using C57B1/6 X C3H mice and S. typhimurium strain TA1535. This activity represented a positive response in 2 of 5 animals tested. Murine testicular DNA synthesis was not inhibited by oral administration of PGE at 500 mg/kg.


Protein methylation in behavioural control mechanisms and in signal transduction. In both prokaryotes and eukaryotes methyl groups can be added to and removed from the carboxyl groups of proteins. Recent work has revealed that these reactions have a role in several behavioural phenomena. The nature of this role has been uncovered in one case--that of bacterial chemotaxis.


The membrane attachment protein for spectrin is associated with band 3 in human erythrocyte membranes. Ankyrin, the membrane attachment protein for human erythrocyte spectrin, is tightly linked in a 1:1 molar ratio with band 3 in detergent extracts of spectrin-depleted membranes. Ankyrin-linked band 3, which represents 10--15% of the total band 3, spans the membrane, and is nearly identical to the major band 3 by peptide analysis. Spectrin binds to solubilised ankyrin-linked band 3, but not to free band 3. A portion of band 3 remains firmly associated with detergent-extracted cytoskeletal proteins. It is concluded that a fraction of band 3 is attached to the erythrocyte cytoskeleton through association with ankyrin, which in turn is bound to spectrin.


The culture of chick embryo dorsal root ganglionic cells on polylysine-coated plastic. Polylysine-coated culture surfaces are strongly adhesive for neural cells, restrict locomotion on nonneuronal elements, but do not inhibit neurite elongation. In the present study, culture dishes were pre-treated with poly-D-lysine (PDL) at various concentrations, seed with dissociates from 8-day chick embryo dorsal root ganglia, and incubated under conditions that normally support both neuronal survival and nonneuronal proliferation. Pretreatment with low (0.1 mg/ml) PDL concentrations had no effect on neuronal survival and neuritic growth, but entirely prevented an increase in ganglionic nonneurons, yielding a numericallly stable culture greatly enriched in neurons. Higher PDL concentrations caused increasing losses in both cell classes. The 50% levels of cell loss were achieved at about the same PDL dose, but earlier for neurons that nonneurons and still with no impairment of neuritic growth from the surviving neurons. A procedure was developed to compare acid-soluble and acid-precipitable accumulation of radioactivity under 1-hr pulses of [3H]uridine, which was applicable even to poorly attached cells. The cytotoxic effects of higher PDL pretreatments was revealed as early as 6 hr after seeding by 2- to 4-fold lower radioaccumulation. The data are discussed in terms of possible regulations of cell permeability and metabolism by adhesive interactions between cells and their substratum, or other cells.


Role of RNA and protein synthesis in memory formation. A brief review is given of experiments which are concerned with the hypothesis that brain RNA and protein synthesis are directly involved in the establishment of long-term memory. It is concluded that these experiments neither support or refute this hypothesis. A convincing demonstration is lacking of interanimal memory transfer by injection of macromolecular extracts. The majority of experiments which attempt to correlate increased macromolecular synthesis with learning use radioactive precursor methods and these studies do not exclude possible changes in precursor specific activity as the cause of the increased labeling. Although some studies find directly observable changes in brain macromolecules in response to training, their relationship to memory formation is unclear. It is possible that these changes represent only an enhanced production of constitutive macromolecules in response to an increase in cerebral metabolism during training, rather than molecular changes that are directly involved with modifying synaptic connectivity. Inhibitors of cerebral protein synthesis block memory formation, but these drugs are not pharmacologically specific and this complicates the interpretation of these studies.


Effect of barbiturates on release endogenous amino acids from rat cortex slices. The potassium-stimulated, calcium-dependent release of endogenous GABA and glutamic acid was suppressed by pentobarbital. The ouabain and veratridine-stimulated fluxes of the amino acids, calcium-independent processes, were not suppressed by pentobarbital. Release of GABA and glutamic acid was not suppressed by pentobarbital in the presence of the calcium ionophore A23187. Of eight barbiturates studied at equimolar concentrations six were found to inhibit GABA release. Thiopental was the most potent, and phenobarbital and secobarbital were inactive.


Effect of lesions in the suprachiasmatic nucleus-retrochiasmatic area on the inhibition of pulsatile LH release induced by electrical stimulation of the midbrain dorsal raphe nucleus. The present studies were designed to determine if the inhibition of pulsatile luteinizing hormone (LH) secretion reported to occur during midbrain dorsal raphe nucleus (DRN) stimulation involved some portion of the suprachiasmatic nucleus(SCN)-retrochiasmatic area (RCA). In animals having lesions that completely destroyed the SCN but did not extend into the RCA, dorsal raphe-induced inhibition of pulsatile LH release was still present. If, however, a significant portion (more than 40%) or all of the RCA was encompassed by the lesion (with variable or no SCN damage), the inhibition induced by DRN stimulation was prevented. This indicates that the SCN is not necessary for the inhibitory effect of DRN stimulation on pulsatile LH release to occur. However, the pathway responsible for this inhibition does appear to project to or through the RCA. Furthermore, pulsatile LH release was reduced, but present, during the control period prior to stimulation in rats with SCN lesions (and no damage to the RCA), and mean blood LH levels for this group were significantly decreased during the control bleeding period. Therefore, the neuronal pathway activating episodic LH release involves, but is not restricted solely to, the SCN.


Prostaglandins and the release of LHRH from hypothalamic synaptosomes. Effects of indomethacin pretreatment. The possible role of prostaglandins (PGs) in the hypothalamic control of ovulation has been investigated by studying the effects of indomethacin (ID) and PGs on the release of luteinizing hormone-releasing hormone (LHRH) from hypothalamic synaptosomes. Female rats were chronically pretreated with ID on diestrus 2 (D2) and proestrus (P), or on diestrus 1 (D1) and D2. Basal LHRH release from hypothalamic synaptosomes prepared from rats pretreated on D2/P was reduced to 48% after a 20-min incubation period, compared with sham-treated animals. ID pretreatment on D1/D2 caused a similar but less marked reduction in LHRH release (to 66% of controls at 20 min). Extraction of the LHRH remaining within the synaptosomal pellet showed that ID pretreatment also reduced the synaptosome content of LHRH. (15S)-15-methyl PGE2 (15-E2) (10(-4) and 10(-6) M) added to the incubating medium stimulated the release of LHRH BY APPROXIMATELY 40%, but this effect was only observed in synaptosomes prepared from ID-treated rats. PGE2 (10(-4) and 10(-6) M) had no significant effect in either sham-treated or ID-treated groups.


Adult dementia: history, biopsy, pathology. The historical events in the evolution of Alzheimer's disease are reviewed, including the initial description by Alois Alzheimer and the subsequent controversy regarding the nosological specificity of this entity. The similarity of senile dementia and Alzheimer's disease is emphasized. The basis for the modern concept of Alzheimer's disease as premature or accelerated aging is included in the review. The pathological correlates of the major categories of adult dementia have been described. The traditional criteria of neurofibrillary tangles and senile plaques have been re-evaluated using the current insight into these changes afforded by electron microscopy and biochemistry. The significance of amyloid has been described because it occurs within the senile plaque and also as the essential component of congophilic angiopathy. The new information regarding neuronal cell counts and the loss of choline acetyltransferase has been evaluated in terms of an indication of a pathogenic mechanism of Alzheimer's disease. The current understanding of normal pressure hydrocephalus, Creutzfeldt-Jakob disease, and multi-infarct dementia has been described. Brain biopsy in dementia has been described as having diagnostic, research, pathogenic, and prognostic value. The precautions involving the performance and handling of the biopsy have been stressed, particularly because these procedures involve conditions of possible slow virus etiology. The polemic for Alzheimer's disease as aging or slow virus infection has been summarized. At this time a consideration seems justified that Alzheimer's disease is an age-related, slow virus disease due to a hitherto unknown immune defect. Aging as an etiological agent must be clarified before Alzheimer's disease, in any form, can be considered to be an inevitable consequence of longevity.


Meige disease: striatal dopaminergic preponderance. A dopamine agonist (apomorphine) and a cholinomimetic drug (physostigmine) were administered to five patients with blepharospasm and oromandibular dystonia (Meige disease). The effects of haloperidol and levodopa were also assessed. Apomorphine lessened and physostigmine aggravated the facial dyskinesias in all patients, while placebo injections had no consistent effect. Levodopa did not modify the symptoms, but haloperidol attentuated the facial dystonia. Dysfunction of the basal ganglia, characterized by a state of striatal dopamine preponderance, probably underlies the dystonic spasms in Meige disease. The prominent cholinergic enhancement of facial dyskinesias may distinguish this disorder pharmacologically from tardive dyskinesia, a differentiation which has practical therapeutic implications.


Tremor due to sodium valproate.

Tremor due to sodium valproate. Four patients developed postural tremor after ingestion of sodium valproate. The tremor was recorded by a variable-capacitance transducer and was of the "benign essential" type. The dosages of sodium valproate varied between 1000 mg and 2000 mg daily and serum levels were between 34.9 microgram per milliliter and 154.3 microgram per milliliter. Tremor was ameliorated in two cases when the dosage was reduced. In only one case was the serum level in the toxic range for our laboratory. The pharmacology of essential tremor is unknown; production of a similar tremor by a drug could serve as a biochemical model.


[Preliminary experience in extracorporeal renal surgery and autotransplants]. Successful management of stenosis of a polar branch of the renal artery by means of autotransplantation is reported. The main indications for this technique are described: arteriosclerosis and injury to the artery, aneurysm, various forms of ureteral injury, bilateral or unilateral renal tumour, coralliform intraparenchymal lithiasis, together, with the techniques for extracorporeal perfusion of the kidney and the mechanisms of the more commonly adopted conservation solutions.


[Gastrointestinal localizations of malignant lymphomas]. Lymphoma of the alimentary canal is somewhat rare. It has no unusual features and few characteristics, with the result that its clinical, radiological and endoscopic diagnosis is problematic, since its lesions may readily be confused by other neoplastic or inflammatory forms. The difficulty is, of course, primarily felt in the case of primary sites, since the co-existence of other lymphomatous tissues facilitates diagnosis. It is suggested that surgeons should be aware of the existence and treatment of these forms. 13 cases collected from 25 laparosplenectomies for staging since 1971 are presented. Four patients had primary lesions, whereas generalised lymphoma was present in the remainder. Nine patients are still alive after treatment.


[Primary tumors of the spleen].

[Primary tumors of the spleen]. Four cases of so-called primary tumour of the spleen are discussed, stress being laid on their rarity and clinical course. The importance of splenectomy for precise histological diagnosis and assessment of the stage of the disease, factors which are indispensable for the oncologist when planning therapy, is also emphasized.


[A new approach to immunotherapy: the transfer factor]. The transfer factor is a tiny molecule capable of transferring the function of the T lymphocytes (immunological memory and retarded hypersensitivity) from a sensitized to a non-sensitized individual. The exact structure and action modalities of the molecule have not yet been precisely established. The difficulties involved in the study of the transfer factor are aggravated by the lack of any suitable experimental model. The attention of immunologists is attracted by this factor which opens up new prospects for the treatment of cancer, immunological deficiencies and certain infectious and autoimmune diseases. More profound research would appear useful to evaluate if and in what cases a potentiation of the immune mechanism can represent an alternative to immunosuppression.


[Incidence of thyroid diseases in primary hyperparathyroidism]. The incidence of thyroid diseases was evaluated in patients with primary hyperparathyroidism subjected to parathyroidectomy. Eleven patients (26.8%) were affected in this way: 2 with carcinoma, 5 with nodular goitre, and 4 with adenoma. The possible reasons for associations of this kind are discussed, and it is suggested that their high frequency points to a relationship of cause and affect.


[Pharmacokinetic and clinical research of a new phenyl ester of carbecinillin: carfecillin]. Following a review of the latest methods of treating infections of the urinary ways, the pharmacokinetic and clinical results of a new antibiotic, Carfecillin, are reported. The examination was carried out on a total of 10 volunteers. The substance showed good serum levels and fair renal excretion with both the 250 mg and the 500 mg p.o. doses. The clinical study was carried out on a double blind basis versus a standard reference (indanyl ester of carbenicillin) in 26 patients and showed the electivity of the antibiotic in the oral treatment of urinary infections under well tolerated conditions.


[Target cimetidine treatment of peptic ulcer. Considerations and assessment of a new dosage]. 32 patients suffering from peptic ulcer have been selected. Of these 12 formed part of a previous series treated with cimetidine and subsequently relapsed; the other 20 were selected on the basis of endoscopic and functional examinations and were treated for the first time with cimetidine. The drug doses used were 1.2 g for 30 days and 0.3 g for a further 60 days. The purpose of the study was to check for possible recurrences by comparing the results obtained and their maintenance in the long term using these doses. 2 months after suspension of maintenance treatment there has not been any recurrence of the acute pain and dyspeptic symptomatology.


Comparative distribution of LH-RH and somatostatin in the supraoptic crest (OVLT) of the rat. Adjacent serial sections of the organum vasculosum laminal terminalis (OVLT) of the rat were incubated with antisera of luteinizing-hormone--releasing-hormone (LH-RH) and somatostatin and stained by the peroxidase-antiperoxidase technique. The distribution patterns of the two oligopeptides show distinct differences, as already found in the median eminence. A denser, more centrally placed somatostatin deposit is surrounded by a more extensive, looser, more peripheral one of LH0RH. These differences in distribution may reflect an anatomical basis for the differing effects of various manipulations on the OVLT and median eminence stores.


Detection of catecholamine and luteinizing hormone-releasing hormone (LH-RH) containing nerve endings in the median eminence and the organon vasculosum laminae terminalis by fluorescence histochemistry and immunohistochemistry on the same microscopic sections. Distribution of catecholamine (CA) and LH-RH nerve endings in the median eminence (ME) and the organon vasculosum laminae terminalis (OVLT) of the rat was investigated by application of fluorescence histochemistry and immunohistochemistry on the same sections of the tissue. In the ME, those two kinds of endings coexisted in the lateral portion of the middle part of ME, and in the wall of tuberoinfundibular sulcus, where they might be considered to have functional correlation. In the OVLT they were also distributed in fairly near distance, but they were not so closely associated as observed in the ME.


In vitro effects of a synethetic enkephalin analogue (D-Met2,Pro5)-enkephalinamide on the hypothalamus-pituitary--testis function in rats. The effects of (D-Met2,Pro5)-enkephalinamide, a synthetic analogue of the natural opioid pentapeptides, were investigated on the hypothalamus-pituitary-testis axis using in vitro tissue and isolated cell incubation systems. (D-Met2,Pro5)-enkephalinamide in a concentration range 10(-7)--10(-5) M did not alter the testosterone production and the troph-hormone sensitivity of the isolated Leydig cells, or the gonadotroph hormone release and the luteinizing-hormone--releasing hormone (LH-RH) sensitivity of the isolated anterior pituitary cells. However 10(-5)--10(-5) M synthetic enkephalin analogue induced a dose-dependent inhibition of LH-RH content and release of the mediobasal hypothalamus, which was prevented by equimolar concentrations of naloxone.


Further studies on the immunohistochemical localization of alpha-MSH in the rat brain. Using two different specific antisera directed against alpha-melanocyte-stimulating hormone (alpha-MSH), the immunoperoxidase localization of alpha-MSH was performed on sections of the rat brain. With one antiserum, some nerve fibers stained for alpha-MSH were seen close to the apex of ependymal cells near the lumen of the lateral and third ventricles. With the other antiserum, alpha-MSH positive nerve fibers were localized in the pai mater. These results indicate that two different specific antisera directed against a same nolecule can probably recognize different antigenic sites of this molecule. Our findings also suggest a possible source of release for the alpha-MSH present in the cerebrospinal fluid (CSF).


Immunofluorescent localization of microfilaments in neuronal and glial primary cell cultures with antibody against adrenal medullary myosin. The localization of myosin was studied in rat neuronal and glial cell maintained in primary culture, using the double antibody immunofluorescent method. Antibodies were raised against myosin purified from bovine adrenal medulla. Myosin-specific immunoreactivity was found in the cell body and neurites of neuronal cells and in the cytoplasm of glial cells. In the former no typical substructure was observable, whilst in the latter myosin-rich filaments were found forming either a cage entrapping the nucleus or as long cables in cellular morphogenic expansions.


Monoamine neuron regulation of LRF neurons innervating the organum vasculosum laminae terminalis and median eminence. The LRF content of the median eminence (ME) and organum vasculosum laminae terminalis (OVLT) was analyzed following lesions destroying the serotonin neurons of the midbrain raphe and the dopamine neurons of the zona incerta. Six weeks after zona incerta lesions the LFT content of both OVLT and ME was significantly reduced. Raphe lesions did not significantly reduce LRF content of the ME or OVLT. No changes in circulating LH content were noted after either lesion. These observations suggest that the dopamine neurons of the incerto-hypothalamic system may be involved in the regulation of LRF synthesis or release from LRF neurons innervating the ME and OVLT.


Musculocutaneous flaps in reconstructive pelvic surgery. Gracilis musculocutaneous flaps are useful in reconstruction of many crippling pelvic defects. They are especially beneficial for vaginal reconstruction at the time of pelvic exenteration. Twenty patients having vaginal reconstruction with musculocutaneous flaps are presented. Nineteen (95%) have vaginas adequate for sexual function.


Treatment of premature labor with ritodrine: a randomized controlled study. Twenty-nine women with premature labor were randomly assigned to a ritodrine (N = 14) or placebo (N = 15) drug group. The 2 groups were of similar age, parity, weight, gestational age, and cervical change at the onset of treatment. They were treated sequentially with intravenous, intramuscular, and oral drugs and monitored carefully during therapy. There was a significant increase in both the maternal and fetal heart rates during ritodrine treatment, and also a significant decrease in maternal blood pressure. Ritodrine-treated women often complained of palpatations. There was no significant difference in the extension of pregnancy, birth weight, or infant survival for the ritodrine group. Although the 1-minute Apgar scores in the ritodrine group were higher, the 5-minute scores were similar in both groups.


Identification of rhinopharyngeal plasmacytic tumours by immunofluorescence on paraffin-embedded sections. Solitary myeloma is an uncommon lesion which may occur in many organs, mainly in the rhinopharynx. This lesion usually appears as non-secretory, and it is constituted by either typical plasma cells or large and atypical cells resembling epithelial anaplastic cells. Therefore, and immunofluorescent method on paraffin-embedded sections may be applied to reveal the presence and the monoclonality of the intracytoplasmic Ig in the pathological cells. In this paper we report a study on 22 patients suspected to bear a rhinopharyngeal tumor investigated by immunofluorescence. Moreover, in 4 cases peripheral blood lymphocytes were also studied. The results are discussed on the basis of the modern immunological trends.


[Potential transformation of the anterior corneal epithelium of the common frog]. The possibility of transformation of the corneal anterior epithelium in the lens following its separation from the stroma was studied. The corneal epithelium was implanted into: a) empty eye orbit and b) cavity of lensless eye of the Rana temporaria tadpoles. In the eye orbit it continued, as in the normal development, to form the basal membrane. Although in the eye cavity the structures similar to lentoids arose but the specific lens proteins were shown to be asbent from them using immunofluorescence. In both the cases, thus, no transformation of the corneal epithelium in the lens was observed. The role of stroma in the stabilization of differentiation of the corneal anterior epithelium is discussed. It is suggested that the absence of increase in the mitotic activity is one of the causes of failure of the corneal epithelium transformation in the lens.


Lateral orbitotomy.

Lateral orbitotomy. The modified Wright technique for lateral orbitotomy is described, based on 57 orbitotomies. This allows for excellent exposure of the orbit for an area lateral to a line joining the superior orbital foramen to the inferior orbital foramen.


Double wedge resection for severe involutional ectropion. Although the individual elements of our technique are well known to ophthalmic plastic surgeons and ophthalmologists, their combination into one procedure for use in severe cases of involutional (senile) ectropion has not been described. Some authors have advocated performing a two-stage procedure for a severe medial ectropion. We have met considerable success in our one-stage procedure and advocate its adoption.


Tissue adhesives: new perspectives in corneal perforations. In corneal perforations associated with extensive progressive corneal disease, a technique using tissue adhesive closure of the perforation site and reformation of the anterior chamber before penetrating keratoplasty or conjunctival flap mobilization has been described. Over the past year, six eyes have been successfully treated in this manner. This method allows reformation of the eye under local anesthesia as an emergency procedure without incarceration or injury of the intraocular contents and without pain to the patient. The definitive surgical procedure can then be performed safely under retrobulbar anesthesia.


Acute pulpal-aleveolar cellulitis syndrome. I. Clinical study of bacterial isolates from pulps and exudates of intact teeth, with description of a specific culture technique. A wide spectrum of microbes has been demonstrated as etiologic factors in the severest form of acute pulpal-alveolar infection; acute cellulitis. An appreciation of the pathogenic potential or oral microorganisms has been gained. Streptococci have been substantiated as the major group of microbes associated with acute odontogenic infection. Anaerobes, primarily gram-negative, are also significant factors. A culture technique for specific microbial growth has been demonstrated and discussed. A simple and economical procedure has been suggested for transport of culture and office study.


Suturing technique in facial soft tissue injuries. This article has presented suturing techniques and instrumentation and suggestions that we have found to be helpful in the management of facial soft tissue injuries. The reader is encouraged to practice suture and knot tying techniques on fresh pigfeet, which are readily accessible and inexpensive and approximate the characteristics of human skin.


Dental dolorimetry for human pain research: methods and apparatus. Electrical stimulation of human tooth pulp provides a means of safely producing human pain in the laboratory. This paper describes a dolorimetry and data collection system for stimulating volunteers, recording responses and analyzing data. The system allows multilevel stimulation in pseudorandom sequences and analysis of results using the methods of Sensory Decision Theory. It consists of modified commercial equipment, specially designed circuitry, an interface, and a programmable calculator. Fundamental problems and safety considerations for electrical dental stimulation are reviewed. Reliability of stimulation and response measurement is discussed.


Myocardial electrophysiology: intracellular studies on heart cell cultures from newborn rats. Electrical properties of cultured newborn rat heart cells are investigated by the use of microelectrophysiological methods. Amplitudes of resting and action potentials appear close to those of in situ heart cells. Elevated spike rate of rise reveals functional fast sodium channels. An inconstant ratio of cells exhibit pacemaker-like activity but no relationship can be established between this automaticity and the tissular origin of the cultured cells. The pulsation rate appears to be linked to the action potential duration and to the pace-maker potential slope. Spontaneous arrhythmias may occur; they are mainly caused by anomalous conduction and (or) erratic pacemaker driving. Thus heart cell cultures may be considered as a precious tool in the field of the cardiac electrophysiologal and physiopathological studies.


Transmembrane calcium movements and excitation-contraction coupling in myocardial cells. It has been realized for a century that Ca2+ is important in the initiation and the control of mechanical activity. The present paper does not cover the field of excitation-contraction coupling in heart but mainly reports some modern and controversial aspects about the regulation of the internal Ca concentration by the sarcolemma while the role of internal stores is only discussed. The first part deals with flux experiments; the second with the slow inward current mainly carried by Ca ions. An analysis of the mechanical activity reveals that only a part of tension is triggered by this slow current; besides, a second component of tension is demonstrated and an exchange mechanism of Na and Ca ions is described in detail. This countertransport, during depolarization, facilitates an influx of Ca ions (coupled to an efflux of Na ions). During hyperpolarization, or even at the resting membrane potential, it promotes an efflux of Ca ions. Thus, the same mechanism may account in part for the development or for the relaxation of tension according to the membrane potential.


Glucose and palmitate metabolism by beating rat heart cells in culture. The metabolism of glucose and palmitate by new born rat heart cell in tissue culture was investigated to ascertain the usefulness of this model system for metabolic studies. Heart cells in culture oxidize palmitate at a rate which is close to the rate reported in vivo. The fate of glucose is shown to differ from in vivo. High rates of lactate release are observed although there are many evidences that cells are not hypoxic. This impaired regulation is likely due to an impaired uptake of glucose.


[Action of erucic and palmitic acids on rat cardiac myoblasts in primary cell culture. An ultrastructural study (author's transl)]. Primary cultures of beating myocardial cells of neonatal rat are taken in order to observe the ultrastructural modifications caused by certain long chain fatty acids (erucic acid C22 : 1 and palmitic acid C16 : 0). Reference cultures are established and observed at the same time as the others. The eurcic acid create an intense steatosis, on the opposite palmitic acid does not. On the contrary the transormations of certain cellular organites such as mitochondria, dictyosomes, rough endoplasmic reticulum and ribosomes are observed in both cases.


[Effects of alanine on the metabolism of lactate and glucose in the isolated perfused rat heart (author's transl)]. Perfused non working isolated rat heart preparations have been used to study whether alanine (0,89 gl-1) could represent an energy source for the myocardium. Its influence on glucose and lactate metabolism in the heart was also studied. The use of 14C labelled compounds permitted the estimation of the oxidation rate of these three metabolites. The results show that: 1) Alanine was not utilized by the heart. 2) Alanine did not alter myocardial glucose metabolism or lactate oxidation. 3) Alanine lowered lactate uptake by the myocardium which may indicate that alanine influences lactate transport through the cell membranes. One may speculate that in vivo, alanine will not interfere with glucose or lactate metabolism by the heart as, even in acute physiological conditions (exercice, hypoxie, fasting) lactate is always present in blood at higher concentration than alanine.


[Biomedical image analysis by mathematical morphology (author's transl)]. There coexist two types of biomedical image analysers, namely the specialized and the universal ones. This distinction corresponds to the one separating pattern recognition from structural analysis. Only specialized devices can give rise to full automation. Mathematical morphology turns out to be a common theoretical background for both kinds of analysers. It defines the logical conditions according to which quantization is possible, and provides a comprehensive class of algorithms.


[Histomorphometric analysis of bone-marrow adipose density and heterogeneity in myeloid aplasia and dysplasia (author's transl)]. A method for histomorphometric analysis of sternal, or iliac, bone-marrow biopsies is reported. Undecalcified, or decalcified, bone marrow-samples from 169 patients with aplastic or dyserythropoietic anaemia were studied utilizing stereological methods: point counting analysis on serial sections. The most informative structure parameters in each of the marrow were the volume density of adipose cells (after correction for the variation with age) inversely related to marrow cellularity, and the standard-deviation (after angular transformation arc sin square root p) correlated with marrow heterogeneity. Hypercellularity, or heterogeneity, appeared to be related to histo-cytological features of dyserythropoiesis and they were associated with a better prognosis comparatively with hypocellular homogeneous marrows.


[Quantitative study of medullary adipocytes in marrow aplasia (author's transl)]. Quantification of adipocyte surface in relation to hematopoietic tissue was performed quantitatively with a classimat on 21 marrow biopsies from patients belonging to the Cooperative Marrow Aplasia Group. This analysis revealed two quantitative parameters, distribution and homogeneity, which permitted the classification of the biopsies into four groups. Each of these categories has been correlated with clinical and kinetic data.


[Study of the spatial repartition of adipocytes with a texture analyser (author's transl)]. A method is given for quantifying the medullary weakness from biopsies of the iliac bone. It consists in calculating the variogram of adipocyte area proportion in a moving window of 70 micron x 70 micron moving 2 mm along the thin section. A numerical application is provided.


[Quantitative study of the medullar fibrosis in 84 cases of idiopathic myeloid splenomegaly (author's transl)]. 84 bone marrow biopsies were analysed by quantification methods with Classimat. The initial diagnosis was: idiopathic myelofibrosis with splenomegaly. A previous study divided the patients among three categories, according to the classification in three progressive stages of the bone marrow disease. The quantitative lecture was done after reticulin coloration, "in blind", and on several biopsies from the same patient. The results show good correlation with classical histologic methods. The quantitative measure of fibrosis is already important as soon as the first stages of the disease. Correlations with prognosis are more difficult to define.


[Quantitative study of hepatic fibrosis in Hodgkin's disease (author's transl)]. The construction of a prototype apparatus adapted to the constraints of tissue structures has permitted not only the quantification of fibrosis within the liver but also analysis of its distribution. The cumulative histograms thus achieved permit the characterization of the liver in Hodgkin's disease and its differentiation from other forms of hepatic fibrosis.


[Image analysis application at immuno-hematology (author's transl)]. Image analysis is an original method for the determination of blood groups and tissue types and for the serology of syphilis. Major advantages are the rapid and specific discrimination of biological particles present in agglutinated or free form, either fluorescent or stained. The reading of the tests is instantaneous and quantitative. The results are expressed immediately and stored with the use of computers. We have studied three micromethods, two for red blood cell grouping and irregular antibodies screening and one for the serology of syphilis, each one being adapted for an automatic system.


[Gastric mucus glycoproteins : structure, functions and pathology (author's transl)]. O-glycosylated glycoproteins are the main constituents of gastric mucus. They are responsible for its viscosity and for its gel-forming properties. Recent advances accomplished over the last years on its physical and chemical characterization are briefly reviewed. Many issues remain still unsettled concerning the modification of its structure and composition in diseases as well as details of its biosynthesis and its pharmacology.


Quantitative evaluation of the in vitro biosynthesis of gastric mucus glycoproteins. Standardization of the methodology. For the evaluation of a normal or pathological function of gastric mucosa, a reproducible method for the estimation of the biosynthesis of mucus glycoproteins appears to be necessary. Rat gastric mucosal scrapings incorporate in vitro several labeled compounds such as 14C-U-D-glucose, 35SO4, 14C-I-L-fucose and L-G-3H-proline in glycoproteins similar to those synthetized in vivo by native mucosa. The aim of our present work is to describe in detail the procedure we use and to show its reproducibility in 9 separate experiments. Secreted glycoproteins (fraction II) and intracellular glycoproteins (fraction III) obtained during a 4 hours in vitro incubation of rat gastric mucosal scrapings at 37 degrees C in standardized conditions were separately studied. The protein and hexose contents, total incorporated radioactivity and specific radioactivity (cpm/mg protein) were determined in these two fractions. The protein content of fraction II from 2 rat gastric mucosal scrapings incubated in 5 ml was 2 mg +/- 0.4 SEM and its hexose was 38.7 mg per 100 mg protein +/- 3.9. Fraction III contained 6.3 mg protein +/- 1.3 and 15.7 mg hexose per 100 mg protein +/- 3.5. About 27% of the total radioactivity incorporated (from 14C-glucose) was in fraction II (+/- 2.6 SEM) and 73% (+/- 2.6 SEM) in the cell bound fraction III. The distribution of total radioactivity was quite similar to that of total proteins : about 25% proteins were in fraction II and 75% in fraction III with a SEM of about 5 to 10% of the average value. If the biosynthetic activity is expressed as specific radioactivity, i.e. the ratio of incorporated radioactivity to mg proteins, the average for both fraction II and fraction III was about 10,000 cpm/mg protein and the standard error of the mean values are +/- 5.2% of the mean for in vitro secreted mucus glycoproteins (fraction II) and 9.1% in the case of intracellular glycoproteins (fraction III). These values can be considered as a satisfactory index of reproducibility of the method of mucosal scrapings if performed as described above.


Antibiotic-associated pseudomembranous colitis in children. Antibiotic-associated colitis is a rare complication of antimicrobial therapy in children. Ampicillin, penicillin, and clindamycin are the drugs most frequently reported to cause pseudomembranous colitis in pediatric patients. This diagnosis should be suspected in any child with significant diarrhea during or after a course of antimicrobial therapy, especially if the diarrhea persists after the drug has been discontinued. The diagnosis is established by proctoscopic findings of typical plaques of pseudomembranes. Most cases resolve promptly when the implicated antibiotic is stopped; however, the disease can be fulminant, progressing to toxic megacolon, peritonitis, and shock. Therapy of patients who have persistent diarrhea after the offending antibiotic has been discontinued should include oral vancomycin. Close fluid management is crucial for survival.


Neonatal necrotizing enterocolitis: implications for an infectious disease. There is a broad spectrum of presentations and severity of necrotizing enterocolitis. Because it may have several different causes, ncerotizing enterocolitis may be a syndrome rather than a specific disease. The triad of formula feeding, intestinal ischemia, and bacterial growth may be part of the pathogenesis of necrotizing enterocolitis. Bacteria are of central importance for the production of pneumatosis, a prerequisite of which is formula feeding. Bacteria may also contribute to the intestinal injury seen after ischemia. However, the disease in the low risk patient seen during an epidemic associated with a single organism is probably caused by a primary gastrointestinal infection. On the other hand, in the stressed newborn infant with mucosal injury the presence of the appropriate bacteria may be all that is needed to initiate the chain of events leading to necrotizing enterocolitis. Figure 2 illustrates the importance of bacteria in all the causes proposed to be involved in the pathogenesis of necrotizing enterocolitis. Whether bacteria are primary or secondary agents, necrotizing enterocolitis should always be approached therapeutically as an infectious disease.


Infant botulism.

Infant botulism. Infant botulism is a unique neuromuscular disease affecting infants less than six months old. It is the result of intraintestinal toxin production by C. botulinum (toxi-infection). Characteristic symptoms include constipation, lethargy, and decreased feeding. Physical examination often reveals generalized hypotonia with cranial nerve impairment. Recovery is dependent on supportive care in an intensive care setting. The relationship of this disease to the sudden infant death syndrome requires further study.


Yersinia enterocolitica infections in children. Y. enterocolitica has been increasingly associated with a wide range of age-related clinical manifestations in children and adults, including febrile gastroenteritis, pseudoappendicitis, arthritis, sepsis, and focal suppurative disease. Although definite patterns of incidence, prevalence, transmission, and pathophysiology are emerging, much remains to be explained. The alert clinician who notifies his clinical laboratory colleagues that special isolation techniques are required to recover this organism from stool samples, and who submits mesenteric lymph nodes for bacteriologic examination in cases of mesenteric adenitis, will aid attempts to further delineate the significance of this emerging pathogen in the United States. Therapy depends on the form and severity of illness and must be guided by in vitro sensitivity, pending animal and epidemiologic studies.


Imagery and the autonomic nervous system: some methodological issues. The present paper is concerned with the role played by image content in the mediation of autonomic nervous system (ANS) arousal. The minimum methodological requirements of such studies are described including controls for imaging, image content, and expectancy effects. Studies meeting these requirements are then reviewed. It is concluded that image content can be a significant modifier of ANS arousal and that this property is not restricted to images containing affective, e.g., phobic, content. These conclusions have relevance to research into techniques such as biofeedback, Transcendental Meditation, and progressive relaxation, where imagery many have a profound influence but where it has received little direct empirical attentiol.


Do low doses of deoxycholate modify the release of rat jejunal brush border hydrolases? The in-vivo effects of sodium deoxycholate (DOC) at low concentrations on the release of protein and some brush border hydrolases, sucrase (SA), maltase (MA), leucine aminopeptidase (LA), alkaline phosphatase (AP), have been investigated in the rat by a jejunal perfusion technique. During perfusion with DOC (0.125 or 0.25 mmol/l), enzyme release was not enhanced. After removal of DOC from the perfusion solution with 0.125 mmol/l DOC, there was a steady release of SA, MA and AP although enzyme release was increased linearly in the control and the 0.25 mmol/l DOC groups. The results also confirm the deep localization of AP within the membrane.


[Bacteria profiles in four coastal areas of East Zealand--coliform bacteria, E. coli and vibriolike organisms (VLO) (author's transl)]. The prevalence of E. coli, coliforms and Vibrio-like-organisms has been studied at four different sites along the east coast of Zealand. High values were registered in sediment taken from the water-line and at a depth of one meter. Generally there exist significant differences between the bacterial counts of water and sediment, and it is obvious that the greatest concentration is in the waterline sediment. The epidemiological and hygienic consequences of these facts are discussed.


[Initial immunochemotherapy in inflammatory carcinomas of the breast. Preliminary results of a trials in 9 patients (author's transl)]. Traditional treatments have proved to be of little effect on inflammatory carcinomas of the breast. The later show a tendency to early metastases and the mean survival is less than 19 months. The preliminary results of the therapeutic protocol reported here and whose originality lies in the primary use of non-specific immunotherapy and of high-dosage chemotherapy appear to be very favourable. The protocol was used in 9 patients. "Thermographic cooling" was taken as the criterion of operability. Initial chemotherapy was restarted after surgery up to a total of 10 courses, and followed by maintenance chemotherapy for one year. Immunotherapy using BCG was routinely associated with the antimitotic agents. After 41 months, the results were as follows: -- local recurrence after tumorectomy in 2 patients (18th and 31st months); these recurrences were controlled by cobalt. -- the other 7 patients show no sign of recurrence. These 9 patients are presently in complete remission.


[Optimal expiratory positive pressure during artificial ventilation. Application in the treatment of respiratory distress syndrome in the adult (author's transl)]. In a group of 14 patients (7 males and 7 females) ventilated artificially for acute respiratory distress syndrome, the authors defined a level of optimal expiratory positive pressure giving an FiO2 = 1, an arterial pO2 greater than or equal to 400 mmhg and/or an intrapulmonary shunt less than or equal to 15%. Improvement in arterial pO2 under such conditions would appear to be related to maximum alveolar recrutment. This optimal level of expiratory positive pressure would appear to be independent of values of total static pulmonary compliance. The long term use of this technique would seem to be encouraging.


[Cryptococcosis: serodiagnosis by immunoenzymology (ELISA) (author's transl)]. For the first time ELISA test was applied for the human cryptococcosis serodiagnosis. We have used the somatic and metabolic antigene extracted from culture of Cryptococcus neoformans. It seems that metabolic antigenosis more specific and more actif. For two cryptococcosis conformated cases (9 sera), ELISA test was always and intensely positive and, in one case after curative treatment, we have seen an importante antibodies decrease. If this first results were confirmated, the ELISA test should give some new perspectives for the sero diagnosis of the cryptococcosis and for the survey of the treatment, that was until now in practice unrealizable.


[Tubal repermeabilisation after sterilisation. Techniques and results (author's transl)]. Requests for repermeabilisation surgery, previously rare, are becoming more common. The indications for reparative surgery are analysed. Microsurgery is possible when the length of the remaining tube us adequate, more than 5 cm, and the ampulla undamaged. It is made difficult when the initial sterilisation procedure has destroyed too long a segment of the tube: electrocoagulation, resection of a long loop of tube. It is impossible when the ampulla and the adjacent portion of the tube have been damaged. Microsurgical techniques certainly improve the results of reparative surgery. After end-to-end anastomosis of the medial half of the tube, 60 to 75% of pregnancies going to term are obtained, and more than 90% secondary permeability. New methods for surgical sterilisation such as clips or rings should make it possible to achieve even better results in the future.


[Lymphocyte migration inhibition in rabbit experimental syphilis (author's transl)]. During rabbit experimental syphilis a suspension of Treponema pallidum (TP) was used as antigen. The evolution of anti TP antibodies was followed up to the fifteenth month, the antibody titer was maximum at month seven. A stimulation of leukocytes migration test was observed up to the sixth month then a significant inhibition appeared (chi square test) which persists after the ninth month. Hence the possibility was discussed that some form of cellular immunity appears secondary while immune response was mostly of humoral type at the beginning of the disease.


[Radio-immunometric determination of the ferritin for evaluation of the iron storage pool (author's transl)]. Seventy-three patients with several haematological diseases have been studied with radio-iron-kinetic, cyto-chemical measurement of iron in the bone marrow, and radio-immunometric determination of the ferritin in the serum. This method gives results which correlate significantly with other methods which evaluate the iron storage pool, in normal or iron-deficient patients. In some cases, an excess of serum ferritin, contrasting with normal serum iron, is confirmed by cytochemical and/or Fe-kinetic studies. In myeloproliferative diseases however, secretion of ferritin by immature cells may induce an excess of serum ferritin. In such cases, the dosage of ferritinaemia cannot be considered as an index of the iron storage pool. Future development of specific dosage of isoferritins could enable to measure the true iron stores, as well as to give an index estimating the evolution of the abnormal cell population.


[Palliative oesophago-jejunal anastomosis for advanced carcinoma of the stomach (author's transl)]. Palliative oesophago-jejunal anastomosis via a purely abdominal approach is proposed in the treatment of inoperable carcinoma of the stomach, and in which gastro-enterostomy is not possible. The latero-lateral nature avoiding any dissection of the oesophagus and the wide calibre of the anastomosis made possible by automatic suture clamps are factors in ensuring the reliability and effectiveness of the method, which considerably improves the living conditions of these patients.


The nucleotide sequence of the major glutamate transfer RNA from Schizosaccharomyces pombe. The nucleotide sequence of glutamate tRNA1 from Schizosaccharomyces pombe was determined to be pU-C-C-G-U-U-G-U-m1G-G-U-C-C-A-A-C-G-G-C-D-A-G-G-A-U-U-C-G-U-C-G-C-U-U-U*-C-A-C-C-G-A-C-G-G-G-A-G-m5C-G-G-G-G-T-psi-C-G-A-C-U-C-C-C-C-G-C-A-A-C-G-G-A-G-C-C-AOH. The sequence differs markedly from that of S. cerevisiae tRNAGlu. S. pombe glutamate tRNA1 can be aminoacylated by the homologous glutaminyl-tRNA synthetase as well as by the corresponding enzyme from S. cerevisiae.


Propagation of restriction fragments from the mitochondrial DNA of Saccharomyces cerevisiae in E. coli by means of plasmid vectors. Some of the EcoRI fragments of yeast (Saccharomyces cerevisiae) mitochondrial DNA were cloned into E. coli using plasmid pMB9. The five smallest fragments in molecular weight appeared to be preferentially retained by E coli; partial fragments derived from larger mitochondrial DNA fragments were also found. One of the fragments, R7 (2.4 kb), may contain the OII gene. Cloned R7 DNA was stable under a variety of growth conditions, but showed some changes in molecular weight after transfer to different E. coli strains. Fragment R7 is transcribed in minicells, producing RNA that hybridizes specifically to mitochondrial DNA. Both DNA strands are transcribed, in contrast to the asymmetric transcription found in mitochondria. No new polypeptides were observed in minicells containing cloned fragment 7.


Segmental flexibility in Escherichia coli ribosomal protein S1 as studied by fluorescence polarization. Ribosomal protein S1 covalently reacts with approximately one equivalent of iodoacetylethylenediamine (1,5-napthol sulfonate (IAEDANS) or iodoacetylaminofluorescein (IAAF). The product AEDANS-S1 can bind to 30S ribosomal subunits lacking S1 as shown by polyacrylamide-agarose gel electrophoresis AEDANS-S1 and AAF-S1 when added back to S1-depleted 30S subunits modulate poly(U)-dependent polyphenylalanine synthesis in the presence of IF3 in a very similar way to unmodified S1. AEDANS-S1 also stimulates RI7-dependent fMet-tRNA binding to 1.0M NH4C1 washed ribosomes whereas AAF-S1 does not. Both static and nanosecond fluorescence polarization techniques were used to study the rotational motions of AEDANS-S1. Several previous studies had indicated that S1 is a highly extended protein which can be modeled by a prolate ellipsoid with an axial ratio of 10 to 1. However, the rotational correlation time we find is about half that expected for such a particle. This suggests that S1 is a flexible protein with at least two domains that can rotate independently.


A ribonuclease-resistant region of 5S RNA and its relation to the RNA binding sites of proteins L18 and L25. An RNA fragment, constituting three subfragments of nucleotide sequences 1-11, 69-87 and 89-120, is the most ribonuclease-resistant part of the native 5S RNA of Escherichia coli, at 0 degrees C. A smaller fragment of nucleotide sequence 69-87 and 90-110 is ribonuclease-resistant at 25 degrees. Degradation of the L25-5S RNA complex with ribonuclease A or T2 yielded RNA fragments similar to those of the free 5S RNA at 0 degrees C and 25 degrees C; moreover L25 remained strongly bound to both RNA fragments and also produced some opening of the RNA structure in at least two positions. Protein L18 initially protected most of the 5S RNA against ribonuclease digestion, at 0 degrees C, but was then gradually released prior to the formation of the larger RNA fragment. It cannot be concluded, therefore, as it was earlier (Gray et al., 1973), that this RNA fragment contains the primary binding site of L18.


The use of R-looping for structural gene identification and mRNA purification. A method is presented for the purification of mRNAs and the identification of structural gene sequences in recombinant DNA molecules. RNA is hybridized to double-stranded linear DNA such that R-loops are formed between most DNAs and their complementary RNA sequences. These R-loops are purified from unhybridized RNAs by gel filtration chromatography in the presence of a high concentration of salt. The complementary RNAs are released from the R-loops by heating, and are assayed by gel electrophoresis or cell free translation to determine their purity and to identify the proteins for which they code. We have demonstrated that recombinant DNAs containing sequences for abundant or moderately abundant mRNAs of Saccharomyces cerevisiae can be identified by this means.


Improved methods for the formation and stabilization of R-loops. Improved methods for the formation and stabilization of R-loops for visualization in the electron microscope are presented. The two complementary strands of a duplex DNA are photochemically crosslinked once every 1 to 3 kb using 4, 5', 8 trimethylpsoralen. R-loops are then formed by incubation with RNA in 70% formamide at a temperature above the DNA melting temperature. Finally, the R-loops are stabilized by modifying the free single strand of DNA with glyoxal, thus minimizing the displacement of the hybridized RNA by branch migration. In this manner R-loops can be formed and visualized at a high frequency irrespective of the base composition of the nucleic acid of interest.


A study on the unprimed poly (dA-dT) synthesis catalyzed by preparations of E. coli DNA polymerase I. Evidence was obtained indicating that the initiation of poly (dA-dT) de novo synthesis is provided by deoxynucleoside diphosphate: oligonucleotide deoxynucleotidyl transferase (dNDP-transferase present in preparations of E. coli DNA polymerase I and capable of catalyzing the unprimed polymerization of dNDP. dNDP-transferase synthesyzes short oligonucleotides which form template-primer complexes repeatedly replicated by DNA polymerase I. This conclusion was based on the following observations: the abolition of the lag period of poly (dA-dT) synthesis by preincubation of DNA-polymerase I preparations with dADP and dTDP; the presence of oligo (dA-dT) among the preincubation products; the suppressive effect of dithiothreitol and N-ethylmaleimide (inhibitors of dNDP-transferase) on the de novo, but not on the primed synthesis of poly (dA-dT), catalyzed by preparations of DNA-polymerase I.


Rapid chemical synthesis and circular dichroism properties of some 2'-5'-linked oligoriboadenylates. Specific synthesis of some oligoadenylates including A2'p5'A2'p5'Ap(2'), the 2'-phosphorylated oligoribonucleotide core of the recently discovered protein synthesis inhibitor pppA2'p5'A2'p5'A is described using a novel solid-phase method. The CD spectra of A2'p5'Ap(2'), A2'p5'A2'p5'Ap(2') and A2'p5'A2'p5'A (derived by treatment of the phosphorylated synthetic trimer with E. coli alkaline phosphatase) are presented. Comparison of the latter spectrum with that of A2'p5'A2'p5'A obtained similarly from a biologically derived sample of pppA2'p5'A2'p5'A provides further evidence that this molecule is in fact the first naturally-occurring 2'-5'-linked oligoribonucleotide.


Studies on gene control regions X. The effect of specific adenine-thymine transversions on the lac repressor-lac operator interaction. Chemical and enzymatic methods were used to synthesize a transition (AT to GC) and a transversion (AT to TA) at a lac operator site known to interact with lac repressor through the thymine 5 methyl group. These operators also contained a poly(dA) . poly(dT) tail 8 to 12 base pairs in length at one end. Results suggest that the steric constraints of lac repressor relative to the position of the 5 methyl group are quite critical. For example a seven fold reduction in stability was observed for the transversion. Results also suggest that the operator spans at least 21 base pairs.


1H NMR studies on the conformational characteristics of 2-thiopyrimidine nucleotides found in transfer RNAs. The molecular conformations of naturally occurring 2-thiopyrimidine nucleosides (5-methylaminomethyl-2-thiouridine, 5-methoxycarbonylmethyl-2-thiouridine and 2-thiocytidine) and 5'-mononucleotides (5-methylaminomethyl-2-thiouridine 5'-monophosphate and 2-thiocytidine 5'-monophosphate) in 2H2O solution were elucidated by analyses of the proton NMR spin-coupling constant, nuclear Overhauser effect, and lanthanide-induced shifts and relaxation enhancements. As monomers, these nucleotides are almost exclusively in the 3E-gg-anti form, even in the absence of ordinary stabilizing factors of this form; i. e., base-stacking and base-pairing interactions with other nucleotide units. This inherent conformational rigidity of the 2-thiopyrimidine units probably contributes to stability of the conformation of tRNA.


Studies on the RNA and protein binding sites of the E. coli ribosomal protein L10. We have used modification of specific amino acid residues in the E. coli ribosomal protein L10 as a tool to study its interactions with another ribosomal protein, L7/L12, as well as with ribosomal core particles and with 23S RNA. The ribosome and RNA binding capability of L10 was found to be inhibited by modification of one more of its arginine residues. This treatment does not affect the ability of L10 to bind four molecules of L7/L12 in a L7/L12-L10 complex. Our results support the view that L10's role in promoting the L7/L12-ribosome association is due primarily to its ability to bind to both 23S RNA and L7/L12 simultaneously.


Characterization of in vitro transcription initiation and termination sites in Col E1 DNA. Overlapping restriction fragments from the region between the single Eco R1 site and the origin of replication of the plasmid, Col E1, have been utilised as templates in an in vitro transcription assay using E. coli RNA polymerase. Transcription towards the single Eco R1 site is initiated at a point 415 bp to the origin side of that site. In vivo, transcription starting at this point probably produces the mRNA for the colicin immunity protein. Transcription away from the Eco R1 site is initiated at a point 140 bp to the origin side of that site and terminated 30 bp further on. This terminator is probably the point at which transcription of the colicin gene is terminated in vivo. DNA sequence analysis in both these regions demonstrated several similarities to other prokaryotic regulatory regions. 50% homology between the putative immunity promoter and other prokaryotic promoters is apparent, so are similarities in AT-content. Upstream of the ATG start codon the sequence PuPuTTTPuPu and a termination codon (TAA) appear; both are typical of prokaryotic ribosome binding sites. The colicin terminator demonstrated similarities to other rho-independent prokaryotic terminators: a GC-rich region with termination in an adjacent AT-rich region containing T clusters on the non-coding strand. The possible role of initiation upstream from the colicin terminator is discussed.


Entrapment of plasmid DNA in liposomes.

Entrapment of plasmid DNA in liposomes. The entrapment of plasmid DNA (pMB9) and high molecular weight DNA into large unilamellar liposomes is described. The entrapment of DNA is specific and due to encapsulation of DNA into the aqueous compartment of liposomes. The entrapped Dna, resistant to deoxyribonuclease treatment, could be reisolated from liposomes intact and, as has been shown by transformation assay, it remains biologically active. The advantages of our method and possible applications are discussed.


Studies on the binding of the ribosomal protein complex L7/12-L10 and protein L11 to the 5'-one third of 23S RNA: a functional centre of the 50S subunit. The RNA binding sites of the protein complex of L7/12 dimers and L10, and of protein L11, occur within the 5'-one third of 23S RNA. Binding of the L7/12-L10 protein complex to the 23S RNA is stimulated by protein L11 and vice-versa. This is the second example to be established of mutual stimulation of RNA binding by two ribosomal proteins or protein complexes, and suggests that this may be an important principle governing ribosomal protein-RNA assembly. When the L7/12-L10 complex is bound to the RNA, L10 becomes strongly resistant to trypsin. Since the L7/12 dimer does not bind specifically to the 23S RNA, this suggests that L10 constitutes a major RNA binding site of the protein complex. Only one of the L7/12 dimers is bound strongly in the (L7/12-L10)-23S RNA complex; the other can dissociate with no concurrent loss of L10.


Nucleotide sequence of the genes III, VI and I of bacteriophage M13. A DNA region of 2750 base pairs encompassing the genes III, VI and I of bacteriophage M13 has been sequenced by the Maxam-Gilbert procedure. By establishing the nucleotide changes introduced by several amber mutations, the coding region and the regulatory signals of each gene have been deduced. The genes appear to span 1275 base pairs (gene III; mol.wt. 44,748) 339 base pairs (gene VI; mol.wt. 12,264) and 1047 base pairs (gene I; mol.wt. 39,500). Their separating non-codogenic regions are extremely short, namely two and one base pair, respectively. The C-terminal end of gene I, however, intrudes 23 nucleotides into gene IV. From the nucleotide sequence it appears that the minor capsid protein of the phage, which is encoded by gene III, is synthesized in a precursor form containing 18 extra amino acids at its N-terminal end. Furthermore, in this capsid protein two clusters of a fourfold repeat of the sequence Glu-Gly-Gly-Gly-Ser are apparent. Gene VI appears to code for a small, extremely hydrophobic polypeptide. Its total hydrophobic amino acids content of 51% suggests that this protein can only function in the host cell membrane.


Blue dextran Sepharose chromatography of the tryptophanyl-tRNA synthetase of E. coli: a potential application for the purification of the enzyme. E. coli tryptophanyl-tRNA synthetase can form a complex with Blue-dextran Sepharose, in the presence or in the absence of Mg++. In its absence, the complex is dissociated by either ATP or cognate tRNATrp. However, in the presence of Mg++, only tRNATrp can dissociate the complex whereas ATP has no effect. E. coli total tRNA or tRNAMet, at the same concentration, cannot displace the synthetase from the complex. It is suggested that the Blue-dextran binds to the synthetase through its tRNA binding domain. This hypothesis is supported by previous findings with polynucleotide phosphorylase showing that Blue-dextran Sepharose can be used in affinity chromatography to recognize a polynucleotide binding site of the protein. The selective elution by its cognate tRNA of Trp-tRNA synthetase bound to Blue-dextran Sepharose provides a rapid and efficient purification of the enzyme. Examples of other synthetases and nucleotidyl transferases are also discussed.


Comparative histologic, histochemical, and histomorphometric studies of T and B cell areas in peripheral lymphoid organs of normal young adult BALB/C mice. T cell areas can be distinguished from adjacent B cell compartments in routinely stained histologic sections of paraffin-embedded mouse lymph nodes and spleens by their comparatively loose texture. The reliability of this morphologic criterion for identifying T cell areas has been proven in comparative histologic, enzyme-histochemical, and immunohistochemical investigations. It is thus possible to determine the size of functionally different compartments in the lymph nodes and spleen of the mouse with sufficient accuracy by the use of conventional histologic techniques. According to our subsequent histomorphometrical studies in two groups of inbred mice matched with respect to age and strain, the proportions of B cell areas in the peripheral lymphoid organs varied significantly depending on environmental conditions, whereas the proportions of T cell areas were stable.


Histological and immunohistological studies on gastric mucosa. I. The presence of CEA in dysplastic surface epithelium. Surface and foveolar gastric cells (SE) of 74 gastric resection specimens were studied by classical and indirect immunoenzyme histology. Immunoadsorbed CEA-antibodies reacting only with the specific determinant of of the CEA molecule were applied. The following SE types with different CEA distribution patterns were observed: Type SE-O, no CEA, normal neutral mucosubstances (MCS); type SE-A, CEA present in the lower half of the SE, important decrease of MCS, mild cell dysplasia by conventional criteria; type SE-B, CEA present in the total cell, absence of MCS, moderate until marked dysplasia; type SE-C, CEA present or decreased, absence of MCS, very important cell dysplasia resembling cancer cells. CEA proved to be useful as marker for the degree of SE dysplasia.


Chlormethiazole--no hangover effect but not an ideal hypnotic for the young. The hangover effects of 768 mg chlormethiazole, an hypnotic with a half-life of approximately 4 hr, were compared with those of placebo in a double-blind cross-over study in 8 young subjects. Ten hours after dosing there were no differences between the preparations in the subjects' psychomotor performance, EEG sleep scores, or visual analogue ratings. Thus, an hypnotic which is rapidly removed from the body may confer considerable advantages. However, all 8 subjects had unpleasant nasal symptoms following chlormethiazole, and it is therefore not an ideal hypnotic for this age group.


[Caval catheterization via the innominate vein (author's transl)]. Experience gained in 3500 supraclavicular innominate vein punctures for catheterization of the superior vena cava with the indirect technique is explained and discussed. The innominate vein is easily accessible in every state of blood circulation, even intraoperatively when the patient is covered by drapes. The thrombosis risk is reduced because of the wide lumen and the straight course of the vessel (vena anonyma dextra). The special advantages of the indirect technique are the small puncture trauma and absence of false positions. The only important complication observed during an average infusion time of 8 days was pneumothorax at a rate of 1.4%.


Severe bronchial asthma requiring ventilation. A review of 20 cases and advice on management. The natural history and outcome of 20 asthmatics who required mechanical ventilation is reviewed. Seven of these patients died. The majority of the deaths could have been prevented if: (1) The patient had been followed-up in an asthma clinic and had been fully informed of the danger of an asthmatic attack. (2) The medical staff had been trained more fully in the evaluation and management of a severe asthma attack. Careful sequential assessment will enable timely transfer of patients with severe bronchial asthma to a unit experienced in their management.


[Development of anaesthetic technique for endolaryngeal surgery 1960--1976 (author's transl)]. In 1960 direct laryngoscopy in combination with general anaesthesia with relaxation and intermittent positive negative pressure ventilation via a smallbore blocker tube was introduced. When, in 1965, microlaryngoscopy was developed it was exclusively performed with this technique. Since 1960, 44, 464 ear, nose or throat operations were carried out. 3,305 (7.4%) were endolaryngeal operations. 943 of them were performed in surface analgesia. 2,363 microlaryngoscopic operations were done under general anaesthesia. 22.5 per cent of the patients were women and 77.5 per cent were men. Their age varied between 6 weeks and 86 years. 2.4 per cent were children under 6 years of age and 33 per cent were aged over 60 years. The main advantages of this method over "open laryngeal surgery" are: 1. it provides a large measure of safety for the patient since even old and obese persons with a rigid rib cage can be adequately ventilated; the cuff prevents aspiration; there is no danger of the patient waking up during relaxation since he is being kept ventilated with a mixture of nitrous oxide-oxygen and halothane. Ventilation via the blocker tube begins immediately after intubation and not, as in open jet ventilation, after insertion of the laryngoscope. 2. The surgeon and his team are not exposed to the risk of infection since, in contrast to the "open larynx" methods, the closed system effectively prevents the escape of pathogenic micro-organisms.


[A system for "intermitten mandatory ventilation" (IMV) using the engström respspirators ER 200/300 (author's transl)]. At low financial and material costs conventional Engström-respirators (Types ER 200, 300) can be converted to make Intermittent Mandatory Ventilation (IMV) possible. These so converted respirators were examined when a group of patients in a surgical intensive care unit who had undergone mechanical ventilation for a longer period of time was weaned from the respirator. It could be shown that during the step-by-step weaning of these patients no significant changes appeared in the results of the blood gas analyses, which means that the patients' pulmonary function took over again gradually. Doubtlessly, the convertability of the Engström-respirator for IMV will help promote the weaning of patients with respiratory complications by this method.


[Treatment of auto-immune diseases with special reference to their pulmonary manifestations (author's transl)]. The treatment of the Goodpasture syndrome, collagenoses (rheumatoid arthritis, systemic lupus erythematosus, progressive systemic sclerosis/scleroderma, matosus, progressive systemic sclerosis/scleroderma, dermatomyositis/polymyositis), necrotizing angiitis (periarteritis nodosa, Wegener's granulomatosis) and idiopathic pulmonary fibrosis is reviewed. Their pathogenesis and morbid anatomy is outlined. The uncommunness of these disorders and their pulmonary manifestations is emphasized. The most frequently employed therapeutic agent are the corticosteroids together with acetylsalicylic acid and indomethacin. The effectiveness of immuno-suppressive and cytostatic drugs (azothioprine, cyclophosphamide, chlorambucil) and of D-penicillamine in the treatment of the different disorders is described. Standard programmes for treating any of these diseases have not yet emerged; the long-term results tend do be unsatisfactory. Co-operation with other specialists (nephrologists, rheumatologists, dermatologists) is often necessary in the treatment of these disorders.


[A case of extremely severe chest injury with fracture of several ribs (author's transl)]. A 72-years old man was severely injured when a lorry rolled back and pinned him down, causing contusion of the chest, fractures of ribs 3-10 on the right and haemothorax. Treatment of the chest injuries was by drainage and by positive end-expiratory pressure ventilation because of the development of severe pneumonia with wet lung. Persistent renal insufficiency, a gastro-intestinal haemorrhage and diabetes also required treatment. The patient developed septic endomyocarditis as a late complication, possibly attributable to the central venous catheter. All pulmonary and extrapulmonary injuries and complications could be set right during the patient's 4-months' stay in the intensive care unit.


[Pneumothorax and haemothorax (author's transl)]. Pneumothorax and haemothorax constitute serious complications of a blunt chest trauma. During 1970-1978 64 persons with pneumothorax and 109 cases of haemothorax were admitted to and treated in the Surgical Unit of the University Hospital, Göttingen. In nearly every case fracture of several ribs had also occurred. The method of choice for treating cases of pneumothorax was by Bülau drainage; surgical closure of the air leakage was rarely necessary. Conservative measures are indicated only if the pneumothorax is narrow or confined to the apical region. It is worth mentioning that in a large number of cases pneumothorax developed during positive pressure ventilation after a chest injury. Complications of Bülau drainage were: damage to the lungs, skin emphysema and bleeding from the intercostal artery. Open application of the Bülau drainage in the 2nd intercostal space is therefore recommended. Treatment of traumatic pleural effusions by Bülau drainage was required in only 14 of 109 cases. Therapy-resistent effusions necessitated thoracotomy and evacuation of the haematoma in 4 persons. If Bülau drainage fails completely to evacuate the haemothorax, it should be promptly followed by thoracotomy to prevent the development of adhesions and the resulting impairment of respiratory function.


[Diagnosis and treatment of injuries to the tracheo-bronchial system (author's transl)]. Clinical symptoms such as mediastinal and tissue emphysema, impaired inflow with cyanosis and extreme dyspnoea, signs of tension pneumothorax suggest rupture of a bronchus; nonstop loss of air via intercostal drainage is an almost certain sign and bronchoscopy provides the final proof. Injury to smaller bronchi may remain asymptomatic. 1,600 persons were treated for chest injury during the past 17 years, 7 of them on account of a ruptured bronchus. The right and left main bronchus were involved in 4 cases and one case respectively. The bronchus of the right upper lobe and the trachea were ruptured in one case each. Once the injury has been diagnosed surgical repair should follow quickly, although the results of anastomoses performed at a later stage were also satisfactory. Closure was by chromcut knotted sutures. Partial pneumonectomy is indicated only if damage to the lungs is extensive. The postoperative respiratory function of the injured lung was satisfactory in all cases.


[Injuries to the tracheobronchial system (author's transl)]. The primary aim in the treatment of penetrating bronchial injuries must be preservation of the organ. 4 cases of traumatic injury to the bronchi are analysed. If the clinical and/or roentgenological signs suggest involvement of a bronchus the suspicion must be verified by bronchoscopy; if the diagnosis is confirmed, thoracotomy is indicated. In 3 patients the operation was performed within 8 hours of the accident. The bronchial defect was closed by chromcat interrupted sutures. Potential complications and the prognosis of these cases are discussed.


[Results of continuous suction drainage in the treatment of pleural empyema (author's transl)]. 201 adults with non-specific pleural empyema (58 women, 143 men) had suction drainage. Most of the patients were aged 61-70 years. In the 47 patients who had not had antibiotic therapy before admission the main causal micro-organisms were staphylococci, pneumococci and streptococci; in the 149, who had had antibiotic therapy outside, micro-organisms such as Pseudomonas, Esch. coli and Proteus predominated. Suction drainage had to be kept up for a remarkably long time: for 4-8 weeks in 36% of the cases and for more than 8 weeks in 8%. 31 patients died, but only 17 of them as a direct result of the pleural empyema. They had all been "high risk" cases on account of age (average 67.7 years), type of bacteria (59%), serious primary or complicating disease (59%), duration of the pleural empyema (76%). Suction drainage succeeded in 140 patients in clearing the infection; but marked adhesions were demonstrable in 27% of the patients when they were discharged from hospital.


[Standardised antibiotic therapy in major lung surgery (author's transl)]. Sputum and bronchial biopsies and smears were obtained from 221 patients who had undergone partial pneumonectomy without preceding antibiotic therapy. The results of 343 sputum examinations and of 126 bacteriological examinations of biopsies and smears were evaluated. Pathogenic bacteria were demonstrated in about 50% of the sputa and in about 25% of the biopsies and swabs. Anaerobic micro-organisms were extremely rare. Treatment was with tetracycline. Before antibiotic therapy Haemophilus influenzae, staphylococci and E. coli predominated, afterwards there were practically only staphylococci and Esch. coli. The incidence of primary resistance to the usual antibiotics was high, especially in respect of gram-negative organisms. The difference in incidence between primary and secondary resistance was one of degree. The least effective agents were ampicillin and amoxicillin, the most active were the aminoglycosides. Erycin proved satisfactory in infections with grampositive organisms while the cephalosporins were active against both gram-positive and gram-negative bacteria. The development of resistance and the clinical picture suggest that "eradication" of the infection during the postoperative stage is not advisable as it causes the selection of more or less drug-resistant organisms. A more satisfactory approach is gradually to reduce the infection until the immediate postoperative stage is over. Macroscopic and microscopic examination of the sputum is important, as the choice of the appropriate antibiotic is determined by the result of gram-staining.