Formation of NADPH-nitrate reductase activity in vitro from Aspergillus nidulans niaD and cnx mutants.

Mutants of A. nidulans at several loci lack detectable NADPH-nitrate reductase activity. These loci include niaD, the structural gene for the nitrate reductase polypeptide, and five other loci termed cnxABC, E, F, G and H which are presumed to be involved in the formation of a molybdenum-containing component (MCC) necessary for nitrate reductase activity. When forzen mycelia from A. nidulans deletion mutant niaD26 were homogenized in a Ten Broeck homogenizer together with frozen mycelia from either cnxA6, cnxE29, cnsF12, cnxG4 or cnxH3 strains grown on urea + nitrate as the nitrogen source, nitrate reductase activity was detectable in the extract. Similar results were obtained by co-homogenizind niaD mycelia with Neurospora crassa nit-1 mycelia induced on nitrate. Thus, all A. nidulans cnx mutants are similar to the N. crassa nit-1 strain in their capacity to yield NADPH-nitrate reductase in the presence of the presumed MCC. As judged by the amounts of nitrate reductase formed, niaD26 mycelia grown on urea +/- nitrate contained much more available MCC than ammonium-grown mycelia. No NADPH-nitrate reductase activity was found in extracts prepared by co-homogenizing mycelia from all five A. nidulans cnx strains. Wild-type A. nidulans NADPH-nitrate reductase acid dissociated by adjustment to pH 2.0-2.5 AND RE-ADJUSTED TO PH 7 could itself re-assemble to form active nitrate reductase and thus was not a useful source of MCC for these experiments. These results are consistent with the conclusion that the active nitrate reductase complex is composed of polypeptide components which are the niaD gene product, plus the MCC which is formed through the combined action of the cnx gene products. Further, the production of MCC may be regulated in response to the nitrogen nutrition available to the organism.


Genetic and biochemical studies on mannose-negative mutants with colonial morphological alterations.

Four mannose mutants of Saccharomyces cerevisiae are described. In addition to their inability to grow on mannose as sole carbon and energy source, the mutants exhibit a distinct colonial morphological alteration. The isolates form hard colonies when grown on agar and exhibit extreme flocculation in broth. These organisms can catabolize mannose, but only form one-half the mannan found in the wild-type yeasts.


Revertants from RNase III negative strains of Escherichia coli.

E. coli strains carrying the rnc-105 allele do not show any level of RNase III in extracts, grow slower than rnc+ strains at temperatures up to 45 degrees C and fail to grow at 45 degrees C. Revertants which can grow at 45 degrees C were isolated. The vast majority of them still do not grow as fast as rnc+ strains and did not regain RNase III activity. The mutation(s) which caused them are suppressor mutations (physiological suppressors) which do not map in the immediate vicinity of the rnc gene. A few of the revertants regain normal growth, and contain normal levels of RNase III. They do not harbor the rnc-105 allele and therefore are considered to be true revertants. By using purines other than adenine it was possible to isolate rnc + pur- revertants from an rnc- pur- strain with relative ease. They behaved exactly like the true rnd+ revertants isolated from rns- strains at 45 degrees C. A merodiploid strain which contains the rnc+ gene on an episome behaves exactly like an rnc+ strain with respect to growth and RNA metabolis, eventhough its specific RNase III activity is about 60% of that of an rnc+ strain; thus the level of RNase III is not limiting in the cell. The rnc- strains show a characteristic pattern of transitory molecules, related to rRNA, 30S, 25S, "p23" and 18S, which are not observed in rnc+ strains. This pattern is unchanged in rnc- strains and in the revertants which are still lacking RNase III, regardless of the temperature in which RNA synthesis was examined (30 degrees to 45 degrees C). On the other hand, in the rnc+ strains as well as in the true revertants and the rnc+/rnc- merodiploid, the normal pattern of p16 and p23 is observed at all temperatures. These findings suggest that all the effects observed in RNase III- strains are due to pleiotropic effects of the rnc-105 allele, and that the enzyme RNase III is not essential for the viability of the E. coli cell.


Identity of a gene responsible for suppression of aminoacyl-tRNA synthetase mutations with rpsT, the structural gene for ribosomal protein S20.

Sepcialized transducing lines of phage lambda carrying segments between thr and car from the E. coli chromosome have been isolated. With help of these phages it has been shown that the gene sups20 (Böck et al., 1974) corresponds to rpsT, the structural gene for ribosomal protein S20.


Anomalies in the selection of mutants of Schizosaccharomyces pombe resistant to 8-azaguanine.

Anomalies in selection render 8-azaguanine unsuitable as a selective agent for screening forward mutations in continuour cultures of Sch. pombe. This system may, however, provide tha basis for an enrichment method for the simultaneous isolation of mutants at two loci.


Further evidence for the participation of proteins S 3, S 14 and S 19 in tRNA binding to E. coli 30 S subunits.

Previous studies have shown that iodination of 30 S subunits causes inactivation for both enzymatic fMet-tRNA and non-enzymatic phe-tRNA binding activities. This inactivation was shown to be due to the modification of three to five ribosomal proteins [1]. In this report the role of these proteins in tRNA binding activity has been further studied. Purified ribosomal proteins, isolated from modified subunits, are re-assembled into otherwise unmodified 30 S ribosomes and assayed for tRNA binding capacity. The presence of modified S 3, S 14 and S 19 (S 15) in the reconstituted particle results in substantial reduction of both fMet-tRNA and phe-tRNA binding activities. This reduction in tRNA binding activity does not appear to be due to an assembly defect.


Ribosomal protein as substrate for a GTP-dependent protein kinase from yeast.

A protein kinase specific for casein and acidic ribosomal proteins was isolated and partly characterized. It was found that the enzyme utilizes GTP and ATP as phosphoryl donors. Its affinity for ATP was considerably higher than for GTP with the km values of 7.6 X 10(-6)M and 5.5 X 10(-5)M, respectively. Two-dimensional acrylamide gel electrophoresis revealed the phosphorylation of the same ribosomal proteins with either of the [gamma-32P] nucleotides used. It was also shown that one acidic protein (S1 or S2) of 40 S and two acidic proteins (L2 and L3) of 60 S ribosomal subunits were predominantly phosphorylated in vitro. The phosphorylated proteins: L2 and L3 seem to correspond to the proteins of L7 and L12 of E. coli ribosomes. The isolated kinase phosphorylated several basic ribosomal proteins though to a lower extent than the acidic ones.


Catalysis of the peptide bond formation by 50 S subunits of E. coli ribosomes with N-(formyl) methionine ester of adenylic acid as peptide donor.

50 S subunits of E. coli ribosomes catalyze the reaction of the 2'(3')-N-(formyl) methionine ester of adenosine 5'-phosphate and Phe-tRNA resulting in peptide bond synthesis. Cytidine 5'-phosphate stimulates this process on 50 S ribosomal subunits as well as on intact ribosomes. The obtained data show that the areas of the peptidyltransferase donor site which binds the 3'-terminal fragment of peptidyl-tRNA possess completely formed structures on 50 S ribosomal subunits.


A model of promoter recognition along the two helical grooves of DNA.

A speculative model based on published sequences to explain the specific binding of RNA polymerase of E. coli to promoters is suggested: (see article). A fragment 24 base pairs long to the left of the initiation site must not contain the G's in the positions indicated by the circles and must contain T's (or G's) in the positions marked by the squares. In most known cases mutual disposition of the circle and square patterns is as shown above. In one case (the fd G3 promoter) the pattern of squares is shifted by 4 base pairs to the right relative to the pattern of universal non-G's (circles).


[Our method for routine examination of kidney bioptic material by light, immunofluorescence and electron-microscopy].

Authors describe their own method for the investigation of needle-biopsy of kidney using light-, and electron microscopy and immunofluorescent technique as well. Small pieces are cut from both end of the needle-biopsy-cylinder and are embedded for electron microscopy. From the larger central part of it cryostat sections are made (at -20 degrees C) for the immunofluorescent study. Afterwards the piece of tissue remained can be fixed in formaldehyde and embedded in paraffin for light microscopic investigation. All three methods can be carried out even when having a 6 mm long needle-biopsy specimen.


Mutagenic evaluation of ronidazole.

Ronidazole was evaluated for mutagenic potential using in vitro microbial tests and in vivo studies in mice. The microbial test used the histidine requiring mutants of Salmonella typhimurium with and without a rat liver microsomal activation system (Ames test). The studies in mice included the dominant lethal test, micronucleus test and cytogenetic assays. Ronidazole was given orally in doses of 50, 100 and 200 mg/kg/day in the in vivo studies. In the dominant lethal test, groups of male mice were treated for five consecutive days before being mated with untreated females. In the micronucleus test, the mice were administered the compound for 2 or 5 consecutive days; they were killed 6 h after the last dose and bone marrow was examined for the presence of micronuclei in developing erythrocytes. In the cytogenetic assays, bone marrow cells in metaphase were examined for chromosome aberrations, 6, 24 and 48 h after mice were treated acutely with the test compound. In addition, similar examinations of chromosomes were made on mice given five consecutive dosages of ronidazole and killed 6 h after the last dose. The results of the various in vivo studies did not suggest that ronidazole would be mutagenic for the mammal. Ronidazole at concentrations of 10 and 50 mug/plate was found to increase the number of back mutations of missense mutants in the in vitro bacterial test. This finding confirms the results of Voogd et al. [19]. Incorporation of the microsomal activation system had no effect on the mutagenic capability of the test compound. In conclusion, although ronidazole was shown to be mutagenic in in vitro bacterial systems, the in vivo systems did not suggest that the compound would be mutagenic for the mammal.


Mutagenic effect of nialamide on Salmonella typhimurium.

The mutagenicity of an antidepressant drug, nialamide, was studied with Salmonella typhimurium TA1535-8. Nialamied was mutagenic for strain TA1535 in the absence of rat liver extracts.


Mutagenicity of the food colour brown FK and constituents in Salmonella typhimurium.

The food colour Brown FK (EEC Serial No. 124) is a mixture of p-sulphophenylazo derivatives of m-toluylenediamine and m-phenylenediamine and is used in the UK for colouring kippers. Brown FK and its constituents were assayed for mutagenicity in Salmonella typhimurium TA 1535, TA 1537 and TA 1538. Samples of Brown FK from three manufacturers were mutagenic in TA 1538 (framshift mutant) when activated by a rat-liver supernatant fraction. Mutagenicity was linearly dose-dependent in the range of 0--3 mg/plate with activities ranging from 22 to 50 times the spontaneous mutation frequency. One sample of Brown FK was mutagenic in the absence of metabolic activation producing a 16-fold increase in mutation at 4 mg/plate. Two major constituents of Brown FK, 2,4-diamino-5-(p-sulphophenylazo)-toluene (I) and 1,3-diamino-4-(p-sulphophenylazo)benzend (II), each present at about 18% in the complete colour, were mutagenic in TA 1538. Mutagenicity was linearly dose-related in the range 0--1 mumol/plate, with slopes of 0.35 mutants/nmol for compound I and 1.5 mutants/nmol for compound II. This activity was dependent on metabolic activation. Four other major constituents, (di- and tri-substituted diamines) were inactive, as was sulphanilic acid, the major excretion product of Brown FK. The mutagenicity of Brown FK could be largely accounted for by the combined effects of compounds I and II. Earlier studies showed that compounds I and II were responsible for the acute myotoxic effects seen when Brown FK was given per os to rats and pigs. Azoreductive fission of I and II to reactive triamines by gut microflora was thought to be the main metabolic pathway by which Brown FK produced its myotoxic effects, and it is proposed that the mutagenic effects of Brown FK are probably mediated by a similar mechanism.


Mutagenicity of industrial compounds: styrene and its possible metabolite styrene oxide.

Styrene and its presumed metabolite, styrene oxide, were tested for their mutagenic effect on a forward mutation system of yeast and of Chinese hamster cells, and on a gene-conversion system of yeast. Experiments with liver microsomal preparations and host-mediated assay with yeast were also carried out. Styrene oxide was mutagenic in all test systems. Styrene was mutagenic only in the host-mediated assay.


The mutagenicity of benzimidazole and benzimidazole derivatives. VI. Cytogenetic effects of benzimidazole derivatives in the bone marrow of the mouse and the Chinese hamster.

Methyl benzimidazole-2-ylcarbamate (MBC) was mutagenic in mice by the micro-nucleus test. Other benzimidazole derivatives, with the exception of the parent compound of MBC, benomyl, and the very closely related substance 2-benzimidazolylurea, did not produce micro-nuclei in mouse bone marrow. Evidence is presented that MBC acts through inhibition of mitosis and that for this action the carbamoyl group is a necessary but not a sufficient condition. It is also demonstrated that for this particular type of mutagenic activity a threshold limit exists, which seems to be in the order of less than 10 mug MBC per ml blood.


Dominant lethal studies with paraquat and diquat in male CD-1 mice.

The herbicides paraquat and diquat were tested for dominant lethal activity in male CD-1 mice. No mutagenic effects were detected on any stage of spermatogenesis in treated males if these compounds were administered orally up to 4 mg paraquat/kg/body weight/day of 10 mg diquat/kg/body weight/day for 5 days. There was no increase in the number of post-implantation losses as shown by the number of females with one or more early deaths or number of early deaths/pregnancy or the number of early deaths/total implants/pragnancy. There was no evidence of pre-implantation losses as indicated by the total implants/pregnant female. There was no anti-fertility effect on the treated males as measured by the pregnancy frequency or successful mating frequency. The lack of dominant lethal effect was not due to particular insensitivity of the animals used since such effects were amply demonstrated in mice doses orally on 5 days with ethyl methanesulphonate at 100 mg/kg/body weight/day, or intraperitoneally on one day with cyclophosphamide at 200 mg/kg/body weight.


Mutagenicity testing and risk estimation with mammals.

Mammalian test systems are currently used for mutagenicity screening. The necessity and the limitations of standardizing these methods are discussed for dominant-lethal assay. In addition to the refinement of standard methods, the development of new systems in mammals is emphasized. One promising approach is the detection of presumed somatic mutations. Another new development takes advantage of electrophoretic methods for detecting induced structural alterations of gene products. Mammalian experiments will be essential for the assessment of risks from chemical mutagens. The development of standards for the controlled use of chemical mutagens should be guided by the experience accumulated in radiation genetics. Two methods, the measurement of specific-locus mutation rates in mice and the direct determination of the phenotypic damage of dominant genes affecting the skeleton of mice, are recommended for the assessment of the hazard of chemical mutagens.


Practical evaluation of mutagenicity data in mammals for estimating human risk.

Chrosomal aberrations are known to constitute a significant portion of the genetic risk from practically all mutagenic agents. These effects are diverse in both nature and in the stage of the germ cells' life cycle at which they are produced. Arguments are made pointing out many of the problem areas in our understanding the significance of chromosomal aberrations in relation to genetic risk. Data are summarized that offer explanations as to (1) why so few chromosomal effects are recovered after treating spermatogonial stem cells, (2) how chromosome damage and dominant lethality are correlated when postmeiotic germ-cell stages are treated with MMS, and (3) why so few reciprocal translocations are recovered after irradiation of oocytes.


The utilization of in vitro mutagenesis techniques to explain strain, age and sex related differences in dimethylnitrosamine tumor susceptibilities in mice.

The carcinogen dimethylnitrosamine (DMNA) is known to exhibit a high degree of strain, organ, age, and sex related tumor specificity in mice. Using microbial mutagenesis assay coupled with mouse tissue microsomal enzyme activation systems, evidence has been obtained that demonstrated a close relationship between the level of in vitro DMNA activation to a mutagen and in vivo tumor susceptibility. DMNA activation by liver, lung, and kidney microsomes from several mouse strains was compared by measuring the rate of mutagenic metabolites formed during incubation of the carcinogen in mutation assays using Salmonella typhimurium G-46 as the indicator microorganism.


Mutagenicity and DNA-modifying activity: a comparison of two microbial assays.

Aspects of the Salmonella mutagenesis and Escherichia coli DNA polymerase deficient (pol A1) assay procedures for detecting environmental mutagens are discussed. The chief limitation of the pol A1-- assay involves substances that do not diffuse rapidly in agar. This problem can be overcome by performing the test in suspension. A simple procedure for accomplishing this is described. Although the Salmonella assay is more flexible, under routine conditions it does not respond to several classes of substances which give positive responses in the pol A1- system. For optimal testing, it is recommended that the two microbial assays be used in tandem. The DNA-modifying properties of povidone-iodine for eukaryotic and prokaryotic cells are described. Even though this substance does not display mutagenic properties in the standard Salmonella assay, it does so in suspension culture. The basis of the mutagenic and DNA-modifying properties of povidone-iodine appears to involve the iodination of the cystosine residue of DNA.


Metabolic studies and the evaluation of genetic risk from the viewpoint of industrial toxicology.

In recent years some important industrial chemicals, e.g. solvents, and monomers used in the production of plastics, have been found to be more dangerous than had been suspected. Some of them are mutagens and carcinogens. The active substance may be the compound itself, but more frequently it is a reactive intermediate, an alkylating agent, formed from the parent compound by oxidation under the influence of liver-microsome mixed-function oxidases. Trichloroethylene, vinyl chloride, 1,1-dichloroethylene and 2-chloro-1,3-butadiene can be mentioned as examples, where mutagenic or carcinogenic activity has been demonstrated; similar activity of some others is to be suspected. Metabolic studies may explain unexpected effects or they may allow one to predict such effects. Tests on microorganisms with a metabolic activation in vitro seem to be extremely valuable for fast and efficient screening of man-made chemicals to be introduced into the environment, because such tests combine the use of liver microsomes and bacteria for detection and classification of the effect. Mutagenicity tests in mammals do not lose their usefulness, since thay take into account the distribution of the compound in the body, its transport to the target organ, species-specific differences etc. As exposures of workers to chemicals mentioned above have been quantitatively measured regularly, epidemiological studies should be performed with regard to mutagenic and carcinogenic effects in relation to the degree of exposure. Here is a chance for better understanding of how to extrapolate experimental results to human populations and how to establish safe levels of chemicals in the environment if at all possible and acceptable.


A hierarchical approach to mutagenicity testing and regulatory control of environmental chemicals.

A hierarchical approach to mutagenicity testing and regulatory control of environmental chemicals is proposed. The guiding concepts and principles relating to the testing of chemicals for mutagenicity and evaluation of genetic hazards are discussed.


The capacity of Drosophila for detecting relevant genetic damage.

For the detection and study of mutagenic agents, Drosophila offers many advantages. It is a higher organism with a short generation time that is cheap and easy to breed in large numbers. The simple genetic testing methods provide unequivocal answers about the whole spectrum of relevant genetic damage. A comparison of the detection capacity of assays sampling different kinds of genetic damage revealed that various substances are highly effective in inducing mutations, but do not produce chromosome breakage effects at all,  or only at much higher concentrations than those required for mutation induction. Of the different assay systems available, the classical sex-linked recessive lethal test thus deserves priority, in view of its superior capacity to detect mutagens. Of practical importance is also its high sensitivity, because a large number of loci in one-fifth of the genome is tested for newly induced forward mutations, including small deletions. Drosophila is capable of carrying out the same metabolic activation reactions as the mammalian liver. An additional advantage, in this respect, is the capacity of Drosophila for detecting short-lived activation products, because intracellular activation occurs within the spermatids ans spermatocytes. These properties make the test for recessive sex-linked lethals a useful tool for verifying results obtained in the pre-screening of potential mutagens with fast microbial assay systems. In studies on non-disjunction, detailed genetic analysis of the induced changes is possible, and these may shed light on the mechanisms involved. A new adaptation of the bithorax transvection method by Mendelson permits the recovery of high yields of chromosome aberrations in a fast one-generation test.


Misrepair of overlapping daughter strand gaps as a possible mechanism for UV induced mutagenesis in UVR strains of Escherichia coli: a general model for induced mutagenesis by misrepair (SOS repair) of closely spaced DNA lesions.

It has been previously reported that an inducible form of post-replication repair appeared to be required for UV induced mutagenesis in a uvrA strain of Escherichia coli. It is shown here that the numbers of daughter strand gaps requiring inducible repair were similar to the numbers calculated to be overlapping one another in opposite daughter chromosomes. An estimation of survival with no repair of these gaps resembled the survival predicted with no mutagenesis. It is the thus proposed that inducible post-replication repair causes mutagenesis by the repair of overlapping daughter strand gaps. A general model for induced mutagenesis is presented. It is proposed that (a) some DNA lesions introduced by any DNA damaging agent may be close enough to interfere with constitutive repair replication of each other, (b) these lesions induce a repair system (SOS repair) which involves the recA+ lexA+ and polC+ genes (c) repair and concomitant mutagenesis occurs during repair replication by the insertion of mismatched bases opposite the noncoding DNA lesions.


Effect of excision repair on azide-induced mutagenesis.

Azide mutagenesis was investigated in Salmonella typhimurium and Escherichia coli. Azide was highly effective in inducing his+ revertants in excision-repair deficient (uvrB) derivatives of S. typhimurium hisG46 and in inducing high frequencies of 5-fluorouracil resistant mutants in excision-repair deficient (uvrA) derivatives of E. coli B/r WP2. In excision-repair plus strains, azide was only a marginal or ineffective mutagen, demonstrating that the bacterial excision-repair system could repair nearly all azide-induced damage. This observation suggests that the initial azide-induced lesion causes a major DNA helix distortion recognizable by the excision-repair endonucleases. The presence of recombination deficient (recB or recC) genes in combination with uvrA increased E. coli sensitivity to azide killing, but depressed azide mutagenicity. These results are similar to those reported for UV-induced mutagenesis with the E. coli strains and suggest that post-replication repair might be the error-prone step in the repair process. Azide mutagenesis specificity is, however, unique and different from UV, as demonstrated by inability of azide to revert the ochre try locus in E. coli WP2s. These results show that the initial azide-induced DNA damage is highly specific but different from UV-induced DNA damage. Metabolic inhibitors, similar in action to azide, did not induce mutations in S. typhimurium strain TA1530, a strain highly susceptible to azide mutagenesis, thus ruling out the possibility that azide mutagenesis was due to peroxide accumulation. A mechanism based on in vivo activation of azide to the actual mutagen is proposed.


The relation between repair of DNA and radiation and chemical mutagenesis in Saccharomyces cerevisiae.

The effect of various genes involved in DNA repair functions on radiation and chemical mutagenesis in Escherichia coli is discussed and compared to similar studies done in yeast. Results of the effect of various genes conferring radiation-sensitivity on mutation induction in yeast are presented and related to current ideas of mutagenesis.


The distribution of fetal death in control mice and its implications on statistical tests for dominant lethal effects.

In dominant lethal testing fetal death is generally assumed to follow either a Poisson or binomial distribution. However, both of these models were found to be inappropriate when three large sets of mouse control data and other data sets from the literature were examined. The validity of statistical test procedures based on these inappropriate models was then studied in detail. It was found that chi-square tests (which assume an underlying binomial distribution) may seriously exaggerate the level of significance and hence should not be used. In contrast, the inappropriateness of the underlying Poisson or binomial model appeared to have little effect on the validity of pairwise comparisons by analysis of variance procedures. Unlike chi-square, these procedures regard the pregnant female rather than the individual implant as the experimental unit. However, a statistical analysis of dominant lethal data generally involves more than a series of pairwise comparisons, and it is unclear how an invalid underlying model may affect statistical test procedures in this more complex situation. Moreover, it is difficult to justify the use of statistical models that are demonstrably invalid when a reasonable alternative exists. Thus, until a satisfactory parametric model can be found and appropriate test procedures derived, we prefer to analyze dominant lethal data by non-parametric (distribution-free) methods. Proportion of dead implants per female appears to be a more meaningful measure of fetal death than number of dead implants per female for several seasons which include (1) analyses based on proportions take the total number of implants per female into account and (2) analyses based on proportions make more reasonable assumptions concerning pre-implantation losses and are more powerful when such losses occur. Despite our concern with the appropriateness of the underlying model, in practice we have found few instances in which non-parametric and analysis of variance procedures have led to markedly different conclusions.


Detection of X-ray induced dominant lethal mutations in mice: an in vitro approach.

The expression of dominant lethal mutations induced by 450 rad of X-radiation in the germ cells of male mice was detected by an in vitro analysis. Embryos sired by irradiated males had a higher incidence of developmental failures in early cleavage, at the late morula stage, and at the late blastocyst stage due to dominant lethal mutations than did control embryos. Dominant lethal mutations were induced most frequently in germ cells exposed as early spermatids and spermatocytes and next most frequently in germ cells exposed as sperm. A decrease in fertilizations, presumably reflecting cell killing, was found in irradiated spermatocytes. These observations indicate that in vitro analysis permits a more detailed description of mutations that are expressed as preimplantation failures than does in vivo analysis.


The production of micronuclei from chromosome aberrations in irradiated cultures of human lymphocytes.

A rapid assay for chromosomal damage would greatly speed studies of the mechanism by which chromosomal aberrations are formed. The characteristics of such an assay--micronuclei produced in cultured human lymphocytes--are given here, together with the evidence that the assay accurately measures X-ray-induced chromosomal damage. Micronuclei arise from chromosomal fragments that are not incorporated into daughter nuclei at mitosis because they lack a centromere. In our experiments the response of lymphocytes from different donors was very uniform and agreed well with what was expected from metaphase analysis of aberrations: (1) the increase in micronucleus frequency begins at the time of the first mitoses, 48 hours after the cultures are started, (2) the exponent of the dose response equation (y=kDn) was 1.2 for micronuclei. For one-hit aberrations n=1 whereas for two-hit aberrations n=2. Since two-hit aberrations predominate in these cultures, a value of n= approximately 1.8 was expected if no increase in mitotic delay or cell death occurred at higher doses, and n less than 1.8 if an increase occurred, (3) the frequency of micronuclei was decreased by a factor of about two when the dose was fractionated, as expected when most of the aberrations are two-hit. The rejoining time for four or five donors was between 30 and 60 minutes, (4) the X-ray-induced micronucleus frequency in cells from people with Down's syndrome (trisomy-21) was twice that of control donors as expected from metaphase analysis [22,23]. Since the micronucleus assay reflects the aberration frequencies so well and is so fast, it is suitable for a rapid assessment of chromosomal damage.


Sister chromatid exchange as an assay for genetic damage induced by mutagen-carcinogens. I. In vivo test for compounds requiring metabolic activation.

An in vivo system has been devised in which chemical mutagen-carcinogens injected into an animal induce sister chromatid exchanges that can be observed when the animal's peripheral lymphocytes are subsequently cultured and then stained with the FPG technique. Chemicals requiring metabolic activation, as well as those that do not, produce significant increases in SCE frequency one day after exposure. The frequency then returns to control level within two weeks. This in vivo system is a highly sensitive assay for the ability of chemical agents to damage chromosomes.


Sister chromatid exchange as an assay for genetic damage induced by mutagen-carcinogens. II. In vitro test for compounds requiring metabolic activation.

Sister chromatid exchanges (SCE's) which are easily seen by "harlequin chromosome" techniques can be readily induced in cultured Chinese hamster ovary (CHO) cells by low concentrations of mutagen-carcinogens that do not require metabolic activation. If the cells are simultaneously treated with cyclophosphamide which does require metabolic activation before it becomes mutagenic, and an activating system consisting of an extract of rat liver containing microsomes (S-9 Mix) then numerous SCE's are induced by the compound. This indicates that the induction of sister chromatid exchanges in such cells can be used as an in vitro assay for mutagens that require activation as well as those that do not. The method, which is very simple and quick, is more sensitive than is the usual cytogenetic assay in which chromosome aberrations are assayed.


An investigation into the potential of a mammalian temperature sensitive leucyl-tRNA synthetase mutant for mutagenesis studies.

Reversion of the mammalian temperature sensitive leucyl-tRNA synthetase mutant CHO tsH1 [33] has been investigated to determine its potential for mutagenesis studies. The protocol for a mutation assay using the new class of temperature sensitive conditional lethal mutants of somatic cells is presented. While it requires careful control of temperature during revertent selection it promises to provide a system complementary to the existing mutation assays. UV and EMS mutagenesis of tsH1 has confirmed that approx. 3TD of expression time are sufficient for complete expression of induced mutants at low mutagen doses (80% survival following UV and 40-50% with EMS). At a higher UV dose resulting in only 10% survival much longer expression times were required which cannot be explained by growth delays alone. While the reason for this is unknown it suggests that care must be taken in studies which require high mutagen doses. Representative revertants which were isolated show a range of phenotypes between those of tsH1 and WT. They appear to be a promising source of extragenic suppressor mutants [33] with alterations in functions affecting protein synthesis.


The fluctuation test as a more sensitive system for determining induced mutation in L5178Y mouse lymphoma cells.

Luria-Delbrück fluctuation tests for the determination of the spontaneous mutation rate to ouabain resistance incultured L5178Y mouse lymphoma cells gave values in the range 0.44 to 1.03 X 10(-7) mutants per cell per generation. Addition of very low, non-toxic, levels of EMS (0.1 mM) and MMS (0.012 mM) 30-48 h before plating in selective medium gave a highly significant increase in the number of ouabain-resistant mutants. Methods for the calculation of spontaneous and induced mutation rates are discussed and a method for the computation of induced mutation rates is described. It is suggested that the modified fluctuation test is a simpler and considerably more sensitive assay for mutagens than the conventional experimental design. Some of its limitations are discussed.


[Effects of light on reproductive processes in fungi; hypotheses on the identity of photoreceptors. A bibliographic revue].

The present review describes the effects of light on reproductive processes in fungi, mainly when action spectra are available. The study of these has resulted in three kinds of photoresponses observable in fungi: - responses only to UV light (230-380 nm) - responses to NUV and blue light (300-520nm) - responses to great wavelengths of the visible spectrum. The photomorphogenetic processes on the control of these same photoreceptor pigments are reviewed.


Spontaneous mutation by mutagenic repair of spontaneous lesions in DNA.

Strains of yeast carrying mutations in many of the steps in pathways repairing radiation-induced damage to DNA have enhanced spontaneous mutation rates. Most strains isolated because they have enhanced spontaneous mutation carry mutations in DNA repair systems. This suggests that much spontaneous mutation arises by mutagenic repair of spontaneous lesions.


Effect of alternate-day steroids on renal transplant function. A controlled study.

A randomized, controlled study of the effect of alternate-day prednisone therapy on renal transplant recipients was done on 53 patients followed for 610 patient months. There was no difference between the 17 recipients of related donor organs who received alternate-day prednisone (experimentals) and their 15 controls who received daily prednisone nor between the 11 experimental and 10 control recipients of cadaver donor organs as regards: rejections per patient month, change in serum creatinine, doses of azathioprine or prednisone, or weight. Likewise the controls and experimentals of the related and cadaver groups did not seem different in either their blood pressure change or their tendency to lose their Cushingoid facies. Alternate-day steroids had no obvious effect on the development of posterior subcapsular cataracts in either related or cadaver organ recipients.


Response to parenteral iron with and without androgen therapy in patients undergoing regular haemodialysis.

The effects of intravenous iron, with and without androgens, was studied in a group of 39 patients treated by regular haemodialysis, almost all of whom had normal serum iron concentrations. Intravenous iron 1-2 g every 4 months produced a significant rise in haemoglobin from 6.3 to 7.9 g/dl in 17 female patients with their kidneys in situ (p less than 0.001). In a group of 13 male subjects intravenous iron plus testosterone produced a similar rise in haemoglobin from 6.9 to 8.6 g/dl (p less than 0.001). Nine nephrectomised patients showed no significant response to iron and androgens. There were no fatalities and no serious side effects in 500 courses of intravenous iron.


Studies on erythropoiesis in patients with renal failure and after kidney transplantation.

Ferrokinetic studies were made in 16 patients before and after kidney transplantation. The Fe clearance decreased in 9 of 11 patients, the plasma iron turnover showed no variation in six patients, while it increased in three and decreased in one, the production of hemoglobin increased significantly after transplantation. The nonnephrectomized patients showed more erythropoiesis after transplantation, manifested by higher hemoglobin level, shorter Fe clearance, and lower serum iron. Uremic patients, including the anephric, had significant responses in erythropoiesis both to stimulation by phlebotomy and to inhibition by blood transfusion. After transplantation, however, the response to phlebotomy was greater and the blockade by blood transfusion smaller, than during uremia.


Renal acidification in patients with mineralocorticoid deficiency.

Renal acidification was studied in eight patients with Addison's disease or bilateral adrenalectomy receiving glucocorticoid replacement only and without renal insufficiency (group I); in five patients with hyporeninemic hypoaldosteronism and moderate chronic renal insufficiency (group II), and in nine control subjects (group III). During acidosis minimal urine pH was 5.4 or less in all subjects, but excretion of ammonium and titratable acid was moderately reduced in group I and markedly reduced in group II. Bicarbonate excretion at normal serum bicarbonate concentration was trivial in group I subjects and inappropriately elevated in two group II patients.


[Limits and prospects of gamma angioencephalography].

Among the informations that can be obtained from the gamma-angioencephalography, the two major aspects are limiting high resolution diagnosis:--on one hand, unspecificity of radio-active molecules for the different lesions,--on the other hand, insufficiency of detection possibilities and scintigraphy enhancement. On this last field fast sequential acquistions technics, data's filtering functional imaging bring much better inflamations. Recently, the new detectors with a high resolution power allow these technics to give to the physicians functional datas that can not be obtained with cumputerized tomography.


[Gamma cysternographic data in hydrocephalus and post-traumatic lesions].

With pneumoencephalography and pressure perfusions tests, the gamma-cisternography is a method of choice for establishing a reliable diagnosis of Normal Pressure Hydrocephalus (NPH) which concept was introduced by HAKIM. The gammacisternography allows to establish the diagnosis of NPH when a persistent ventricular filling is present and to find, very often, the etiological factor by showing the absence of radioactivity over the subarachnoïd pathways along the convexities at any stage or delayed and abnormal CSF absorption. The gammacisternography permits also to explore with 111 In DTPA the high risk patients who develop gradually over the course of months and years a NPH after a head trauma, a meningitis, a subarachnoïd hemorrhage or a neuro-surgery treatment. The NPH is present only 12 to 15% of our dementia cases with gait disturbance. Nevertheless, the search for such cases remains worthwhile as the patient may be able to return to a useful life after shunting procedure. The indication for shunting is not possible ot assess on scintigraphic results alone. It must be kept in mind that the abnormalities of CSF flow may be transient, may progress or regress. The shunting may be decided if the scintigraphic pattern remains unchanged and if the clinical symptoms are going worse. The pronostic is better in statistics after ventricular shunting when the NPH scintigraphic pattern is present. The valvo-scintigraphy can also evaluate the function of a shunt during the post-operative period.


[Diagnosis of cerebrospinal fluid leakages by gamma-cisternography].

Cerebrospinal fluid (CSF) leakages either secondary (traumatic) or spontaneous (non-traumatic) are first considered in their frequency and origin. The exact topography of the various meningeal and cranial lesions involved are difficult to assess particularly in the most important groups of persistant traumatic CSF rhinorrhea and recurrent meningitis. Among the various diagnostic approaches, direct observation is always necessary, but of limited value. Standard X-rays must be followed by multidirectionnal tomography (Polytome) and, whenever available, computed tomodensitography of the base of the skull. Brain pneumography provides a thorough setting fourth of the congenital or acquired cerebral lesions as well as the new cranio-meningeal conditions. Difficulties encountered with the techniques of subdurography and Pantopaque injection are underlined. Three radioisotope techniques are considered. 1) The earlier technique of cotton-pledgets only shows the external orifice. 2) The recent proposal of nuclide cranial subdurography is criticized for ignoring the leptomeningeal bag. 3) Radioisotope cisternography (RIC) or gamma-cisternography is described more precisely. It remains the most complete and appropriate method for observing the natural behaviour of the leakage. RIC with fistulography is performed through suboccipital injection of 99mTc-DTPA. RIC provides essential clues on the relative importance of associated dynamic disturbances of the third circulation and morphological changes of its anatomical bed (stenoses and widenings of the ependymal and leptomeningeal spaces). If present, the leakage may be directly shown on the RIC pictures. If rhinorrhea is abundant, there is no difficulty in assessing side and site of the fistula. If rhinorrhea is occult, dubious or intermittent, diagnosis is often difficult. There are also indirect signs of rhinorrhea: leptomeningeal dilatation near a frontal or ethmoidal fracture, contamination of the rhinopharynx, examination of the handkerchieves for presence of radioactive material. Statistical results from the literature are summarized in a table.


[Radioisotopic methods for studying cerebrospinal fluid shunts].

The article presents a general and critical review of the radio-isotopic methods for assessing the function of the surgical shunts of cerebrospinal fluid (CSF). The radio-isotope cisternography may indirectly demonstrate the function of the shunt by the postoperative modification of the eventual ventricular reflux and of the permeability of the subarachnoid spaces on the cerebral convexity. The methods of direct ventricular injection of the indicator permit a more direct assessment of the drainage, as based on a scintigraphy, or on the quantitative study of the cephalic radio-activity, or on the demonstration of radio-activityon a target-organ. The direct injection of a highly diffusible indicator in the valve allows a quantitative measurement of the flow of CSF in the shunt and brings morphological arguments for the localization of the eventual occlusion. The technical aspects of the latter method are critically discussed and illustrated by personal results.


[Comparative experimental study of microsurgical nerve suture. I. Immediate suture and delayed suture].

Impressed by multiplicity of theories and conceptions concerning nerve surgery and by the difficulty to draw grouded conclusions from the different and contradictory work available from the litterature on the subject, the author makes a revision of the biology of the nerve, to be used as a point of departure for investigation. Based on the fact that protheosynthesis capacity of the nerve cell is known to be at its top value during the first week after the lesion of the nerve, the author endeavours to compare immediate suture with delayed suture. This comparative study is based both on electrophysiological and histological data and show clearly enough the real advantage of trying nerve reconstruction as early as possible, to avoid progression of devitalisation and invasion by fibrosis of both stumps. The surgical procedures are made with the help of the operating microscope and the same technique is used in both groups.


Synthesis of LH-RH by rat hypothalamic tissue in vitro: I. Use of a specific antibody to LH-RH for immunoprecipitation.

Following the procedure of Jeffcoate et al. [1974] we have successfully obtained a specific antiserum to LH-RH in rabbits, and are utilizing our antibody to study the synthesis of LH-RH by rat hypothalamic tissues in vitro. Our antibody, used at a dilution of 1:20,000, binds 37% of added 125I-labelled LH-RH, and cross-reacts minimally with thyrotropin releasing factor (TRF; 4.0%), somatotropin release inhibiting factor (SRIF; 0.035%), and melanocyte inhibiting factor (MIF; 0.009%). The antiserum recovers LH-RH added to charcoal-stripped rat serum quantitatively. Serial dilutions of rat hypothalamic extract and varying aliquots of normal rat serum are parallel to the standard curve (synthetic LH-RH: 0-100 pg/tube). Hypothalamic tissue of 4 male rats, bounded by the optic chiasm, mammillary bodies and hypothalamic fissures to a depth of 2-3 mm, was pooled, minced and incubated for 1-3 h in 2 ml of Eagle's Minimum Essential Medium in atmosphere of 95% O2/5% CO2 with 10 muCi of 3H-glycine. Reactions were stopped by the addition of 1 ml 0.1 N HC1. The tissue and medium were homogenized together and boiled for 3 min. Aliquots from the incubates were neutralized, LH-RH levels measured by radioimmunoassay (RIA), and 3H-glycine incorporation determined by immunoprecipitation, using our antibody. 3H-glycine incorporation into presumptive LH-RH increased linearly over the 3 h period. Aliquots of the 3 h incubates were chromatographed on Sephadex G-25 columns (1 x 10 cm) using 0.01 M acetic acid for elution; 1 ml fractions were collected. Synthetic LH-RH was chromatographed in a similar manner, and its elution profile determined by UV absorbance at 280 mm. Aliquots of each fraction of the eluted material from the 3 h incubates were counted in a scintillation counter to determine the elution pattern of the labelled material. Additionally, aliquots from the 1 ml fractions were neutralized and used for immunoprecipitation. Coincident peaks of 3H-label, immunoprecipitable 3H-labelled presumptive LH-RH, and synthetic LH-RH were observed. These results lend strong evidence to support our conclusion that the technique of immunoprecipitation is an efficacious approach to the study of the synthesis of LH-RH by rat hypothalamic tissue in vitro.


Biphasic effect of estradiol on mechanisms regulating LH release in ovariectomized sheep.

Single injections of 50 mug estradiol-17beta (E2beta) into overiectomized sheep caused biphasic changes in plasma luteinizing hormone (LH). An initial 8-h period of LH inhibition was followed by a period (12-20 h after E2beta) of facilitated LH release. Pituitary LH responsiveness to small dosages of synthetic gonadotropin-releasing hormone (GnRH) was tested repeatedly at 2-h intervals during the time periods when plasma LH was inhibited and when it was facilitated. Reduced sensitivity to GnRH (91% decrease) characterized only the initial 4 h of the inhibitory period, suggesting that E2beta suppressed endogenous LH-releasing factor (LRF) during the latter part of the inhibitory period. Hypersensitivity (2-fold increase) to GnRH was briefly observed at the beginning of the period of E2beta-facilitated LH release. This transient and modest hypersensitivity does not completely account for the very large E2beta-induced increases in plasma LH. Therefore, E2beta probably increased endogenous LRF during the period of facilitated LH release.


Effect of testosterone on hypothalamic LH-RH content.

Hypothalamic LH-RH content in male rats is lowered after castration. The s.c. implantation of testosterone or testosterone propionate-packed Silastic tubing (from 0.5 to 6 cm in length) in a range which encompassed the normal circulating plasma testosterone concentration, prevented this lowered LH-RH content 21 days following castration and simultaneous implantation. The temporal response to implanted testosterone was then studied: rats were killed 1,2,4,8,14 and 21 days after castration and simultaneous implantation of 3 cm testosterone-packed Silastic tubing. The hypothalamic LH-RH content began to decrease in the castrated group after 4 days and fell progressively thereafter. However, the hypothalamic LH-RH content of the castrated group maintained with constant levels of testosterone showed no such reduction at any time following castration. These experiments indicate that circulating testosterone in physiological concentrations can maintain a normal hypothalamic LH-RH content and demonstrate an action of testosterone, in physiological concentrations, in the feedback regulation of LH-RH secretion.


Alpha-methyldopa-chlorpromazine combination in schizophrenic patients.

In a double-blind, controlled clinical study, lasting 8 weeks, 55 hospitalized schizophrenic patients were randomly assigned to one of the five drug treatments: 600 mg chlorpromazine/day, 300 mg chlorpromazine/day, 300 mg chlorpromazine--250 mg alpha-methyldopa/day, 300 mg chlorpromazine--500 mg alpha-methyldopa/day, 300 mg chlorpromazine--750 mg alpha-methyldopa/day. The results showed that all five treatments were significantly effective in treating schizophrenic behavior. The dose-response relationship for the different doses of alpha-methyldopa in combination with chlorpromazine, suggested an optimal response to 300 mg chlorpromazine--500 mg alpha-methyldopa.


Neuropsychobiology of affective disorders. Some methodological considerations.

Amine hypothesis is neither an adequate nor a sufficient postulate in understanding the neuropsychobiology of affective disorders. Interactions and balance between various mechanisms and factors may be important. Better techniques and methodology need to be used and a more comprehensive approach seems to be necessary in formulating meaningful research strategies. The present paper reexamines someaspects of the available data and suggests a few approaches in the sutdy and elucidation of neruophychobiology of affective disorders.


[The free autologous dermo-adipose graft in the treatment of postoperative laparocele].

An account of the biological and contentive features of the derma is followed by a proposal for its utilisation in the form of an autologous, free dermo-adipose graft in the management of some forms of post-operative laparocele. Personal experience and technique are explained. Grafts are used both for reinforcement of the wall after simple laparoplasty and in the replacement of missing muscle-aponeurotic planes.


The spread of blood-borne metastases in malignant lymphomas of man.

An autopsy study was performed on 190 cases with Hodgkin's disease and on 134 cases with reticulum cell sarcomas. It was found that a cascade spread of metastases, as opposed to a random spread, was detectable in patients with Hodgkin's disease, but not in patients with reticulum cell sarcoma. Metastatic key-sites for both lymphomas are defined as those organs whose chance of being seeded by the primary tumor is the highest; these sites are the spleen, liver, bone marrow and lungs. This study could not demonstrate in Hodgkin's cases that metastases in the liver and bone marrow were spleen dependent, but rather it showed that the spleen, liver and bone marrow acted as three independent sources of generalized metastases. Explanations proposed to account for the different metastatic spread in Hodgkin's disease and reticulum cell sarcomas, included different intrinsic cellular characteristics of the two lymphomas. the different location of the primary tumor either because of nodal, or of extra nodal origin, or because the primary tumor was located either in the upper or in the lower torso or in both areas. Malignant cells in the kidney, central nervous system and endocrine system were either released by the spleen via the lungs, or released by the liver and the lungs, or they were bone marrow dependent. The cascade spread of metastases hold true only for Hodgkin's disease. In cases with reticulum cell sarcomas, metastases appeared to spread at random from the primary tumor throughout the body, with rare exceptions such as metastases in the central nervous system, which originated either in the liver or in the spleen, lungs, kidney and bones.


Inhibition by ionophore A23187 of the cytotoxicity of vincristine, colchicine and X-rays to leukemic lymphocytes.

The ionophore A23187 was tested for its effect on the sensitivity of normal and leukemic lymphocytes to colchicine, vincristine, and X-irradiation. The ionophore was used because it transfers divalent cations through biological membranes and causes the transformation of normal lymphocytes. The sensitivity of the cells was measured by incubation of lymphocyte suspensions with and without reagent at 37 degrees C for one to seven days and counting the number of viable lymphocytes before and after incubation. Viability of the cells was judged by cytologic criteria as visualized by phase contrast microscopy. Lymphocytes were obtained from patients with chronic lymphocytic leukemia and from normal persons. Colchicine and vincristine killed in one day non-dividing leukemic lymphocytes while normal lymphocytes were much less sensitive to the two alkaloids. X-irradiation (1000 rad) killed nearly all leukemic lymphocytes in two days and normal lymphocytes in three days. Ionophore A23187 (2 X 10(-5)M) was moderately toxic to normal but not to leukemic lymphocytes and caused the transformation of both types of cells. The ionophore inhibited the cytocidal action of colchicine and vincristine to leukemic but not to normal lymphocytes and inhibited the action of X-irradiation to both types of lymphocytes. The present findings and a review of the literature suggest the hypothesis that calcium or other divalent ions are involved in the cytotoxicity of colchicine, vincristine, and X-irradiation of leukemic lymphocytes and that there may be an abnormality of the metabolism of divalent ions in leukemic lymphocytes.


Antibody response to and immunity induced by Corynebacterium pyogenes vaccine.

A procedure is described for producing a high quality C. pyogenes vaccine. A satisfactory antibody response was obtained by administering 3 subcutaneous injections of vaccine at 10-day intervals, using doses of 2,0 ml for rabbits, 5,0 ml for sheep and 10,0 ml for cattle. The titres resulting from this procedure compared favourably with those obtained from following the more intensive schedule and higher doses of vaccine recommended by the British Veterinary Codex. The antibody response was at its best 1-2 months after immunization but dropped to a plateau 6 months after immunization. Mice were also successfully immunized against challenge with 1,5x108 organisms by the application of 3 doses of vaccine administered at 10-day intervals.


[Research on hepatic and overall collagen metabolism in irradiated rats (author's transl)].

The authors studied the effect of ionising radiation on hepatic and overall collagen metabolism in rats. Doses of 100 and 250 rads produced a reduction in urinary hydroxyproline while doses of 500 and 700 rads produced an increase. These results may be interpreted in favour of an increase in reticulation in the first case and in the second case, destruction of inter-molecular bonds. The quantity of total and insoluble collagen increases proportionally to the dose 40 days after the radiation. This proportional relationship is not observed during the first few days after radiation. Prolonged radiation with 700 rads produces a marked reduction in all collagen fractions, compared with 4 days after acute radiation. Among the substances studied, "folcisteine" had the best radio-protective effect when administered before radiation with 700 rads.


[Immunoglobulins in the CSF during meningitis (author's transl)].

Cerebrospinal fluid and serum immunoglobulins were studied in 76 patients with acute meningitis of various aetiologies. This comparative study showed that the increase of CSF immunoglobulin levels was related to important modifications of the permeability of the blood-brain barrier (especially in case of bacterial meningitis) or to a local in situ production (particularly in case of "viral" meningitis). Results obtained allow us to draw a different biochemical pattern in purulent meningitis and acute lymphocytic meningitis.


[Practical interest of a micromethod for neuraminidase treatment of lymphocytes in transplantation immunology (author's transl)].

Vibrio cholerae neuraminidase treatment increases the cell sensitivity to complement and antibodies cytotoxic action. This property can be applied to the microlymphocytotoxicity technic for antibodies study in dialysed and kidney transplanted patients and for pretransplantation cross-matches. The enzymatic treatment usually employed needs a great deal of lymphocytes submitted in a second step to antibodies cytotoxic action. But this method appeared difficult to be routinely applied. We developed a simpler method consisting in treating only the lymphocytes needed to perform the test, on the reaction plate itself. This method gives the same results as the classical enzymatic treatment: the intensity of the weakly positive reaction is strongly increased, after pretreatment of cells; in certain cases, it allows one to detect cytotoxic antibodies not revealed without neuraminidase. These antibodies can be related (or not) to HLA system but practically the more important fact is the ability of this method to reveal an eventual incompatibility.


[Effect of clofibrate on tissue lipase activity in rats].

Given for 20 days at a concentration of 0.3% in the diet, clofibrate lowers serum triglyceride level and causes a 48% increase of lipoprotein lipase activity in rat fat pad, after feeding. This increase does not appear in the animals on fasting. These results agree with an effect of clofibrate upon membranous adenyl-cyclase of adipose tissue.


[Antibody coated bacteria in urinary sediment (author's transl)].

The presence of antibody coated bacteria in the urinary sediment was correlated with the infection site in 151 patients. 86 patients were followed regularly and support sequential data. A positive test is noted in most of the chronic pyelonephritis cases in all the prostatitis cases and in some of the chronic infection on bladder disease. In recurrent cystitis the test is generally negative as in acute pyelonephritis seen within 24 hours.


[Bone marrow's histopathology in tuberculosis of the hematopoietic organs (author's transl)].

Through the reports of the literature, the histopathologic lesions of the bone marrow in tuberculosis of the hematopoietic organs are analysed. Whether the lesions are typical or not, it is necessary to have a bacteriologic proof of tuberculosis. Near these lesions, bone marrow's cellularity is often greater or lower, reticular fibers are often increased and sometimes reticulin fibrosis is marked. Other necrosis and granulomatous inflammation are discussed. Contribution of macrophage is described. The relationships between tuberculosis and hematologic diseases are reported.


Tolerance and leukaemogenesis.

Two subjects are considered here separately. Both are related to findings in the early embryo aggregation derived mouse chimaera model. Such chimaeras are most commonly derived following the aggregation of two undifferentiated embryos and therefore not suprisingly they were originally considered examples of "classic" immunological tolerance. Since this time alternative mechanisms including humoral and cell suppressor activity have been suggested and until recently tolerance in tetraparental chimaeras has remained a controversy. This controversy is now reviewed in the light of recent findings which has suggested that such mice are in fact examples of classic tolerance with the possibility that this is achieved by heterogeneous elimination of the clone of potentially self in equilibrium self auto-reactive cells. Chimaeras have also been studied in respect of leukaemogenesis. Results in a group of AKR reversible CBA leukaemia susceptible reversible resistant chimaeras suggest resistance is dominant. Moreover evidence now points to lack of anti-viral antibody activity in these chimaeras which I wish to suggest may be related to apparent resistance to leukaemia. In this context it may be envisaged that in the absence of masking anti-viral antigen complexes "normal" tumour immunity may have been effected. Although this has yet to be proven evidence points to tolerance to the oncogenic virus being maintained in the chimaeras furthermore also in the naturally derived (AKR X CBA) F1. This in turn leads me to suggest that "intolerance" to the oncogenic virus, the spontaneous development of anti-viral antibodies and tumour development might well be related in the AKR. This in turn enables me to propose that tolerance and leukaemogenesis, at least in this stiuation, appear to be related.


[Bidimensional electrophoresis in the study of cerebrospinal fluid protein (author's transl)].

The authors report 650 bidimensional electrophoreses of the cerebrospinal fluid (CSF). All examinations were carried out after prior concentration of CSF protein. The technic used is that described by Rebeyrotte in 1970. Four types of tracing were noted: -- Two tracings were unusual by the presence of a large peak in the alpha-2 region. -- One tracing was of the hypergamma type. -- One tracing was comparable to serum by the richness of the precipitations. A change in the first migration permitted us to obtain electrophoretic separations from 100 microlitres of pure CSF. The bidimensional immunoelectrophoresis tracings are comparable to those described previously.


[Effect of phosphate upon calcium intestinal absorption (author's transl)].

Adult rats receive 5 to 50 mM CaCl2 solutions in which glycerophosphate or sodium diacid phosphate may be added in variable quantity. These solutions are administered by gavage or in situ ligatured jejunal loop. The inhibition of calcium absorption dependent on simultaneously administered phosphate doses is well characterized: high for the lowest concentration, the inhibiting effect of phosphate doses decreases more and more reaching a limit from which phosphate supplementation has no effect. These observations discarding an intervention of phosphate by calcium insolubilization seem to demonstrate that the control supplied by phosphates on calcium absorption is of enzymatic character. Facts related to the respective effects of calcium and phosphates on the action of alkaline phosphatases lead to discuss a possible intervention of these enzymes upon calcium transfer.


[Bronchial lympho-epithelial nodules in the rat. Definition and morphological characteristics in optical and electron microscopy (author's transl)].

Among the bronchial lymphoid tissue, lympho-epithelial nodules are an original component, easily distinguished from other structures, particularly from lymphoid cell infiltrates. Such structures are encountered in many species and may be characterized by a flattened epithelium, containing a few ciliated cells, devoid of mucus producing cells, and infiltrated with lymphocytes. Inter and intracellular vacuoles are described in the lympho-epithelium and the underlying mucosa. Electron microscope studies support the view that lymphoepithelium has a "trapping" function of particulate matter, more direct evidence is however attempted. Peripheral lymphatic vessels and high endothelium veins contribute to the important lymphocyte transfer. An hypothesis of capacity for antigenic recognition of the lymphoepithelial nodule is discussed.


Antigenic analysis of a variant specific component of Trypanosoma Brucei Brucei I -- Immunochemical purification by affinity chromatography.

An antigenic component specific to the bloodstream form of serotype AnTat-1 of Trypanosoma brucei brucei was purified by immunoadsorption technique. Subsequently, a monospecific rabbit antiserum was prepared. Purity and specificity controls of the final product were carried out by immunoelectrophoretic analysis.


Histological aspects of arterial abnormalities produced by X-irradiation of the aortic and branchial arches in the chick embryo.

Low dose X irradiations centred on one or two aortic and branchial arches of 3-day check embryos result in teratogenic effects on the arteries (21-92%) and in low mortality (4-12%). Histological studies of abnormal arteries show different structural malformations of the arterial wall: inorganized arterial fibres, abnormal intima, newly formed arterial tissue in arterial lumen, coalescence of two arteries. It is suggested that a disturbance at the aortic differentiation level results under our experimental conditions in the observed arterial abnormalities.


[Bactericidal power of polymorphonuclear neutrophils. Trials for standardization of a micromethod using autoradiography (author's transl)].

A 50 mul sample of blood is placed successively in the presence of staphylococci, then of tritiated thymidine. Stripping film autoradiography permits identification of living intracellular bacteria which metabolize the marked precursor. In our experimental conditions, the percentage of marked intracellular bacteria is: -- 15,4 +/- 3,7% (x +/- 1s) among adults -- 14,6 +/- 3,2% among children.


Human platelet fibrinogen: a protein different from plasma fibrinogen.

Fibrinogen has been purified from normal and dysfibrinogenemia Metz plasmas, and from normal and from the patient's platelets. There are differences in the pattern of disc polyacrylamide gel electrophoresis between normal plasma and normal platelet fibrinogens, but the migration of the A alpha chain is similar. The abnormality of the electrophoretic mobility of the A alpha chain of plasma fibrinogen from Metz dysfibrinogenemia is not found in platelet fibrinogen of this patient. This result clearly establishes that platelet fibrinogen is a different protein from plasma fibrinogen.


[Vascular subendothehlium: structure and functions (author's transl)].

Two subendothelial structures, glomerular basement membrane and arterial subendothelium, were compared regarding their biochemistry and ultrastructure and their role in physiology and pathology. Filtration function of the glomerular basement membrane can be related to the presence of three layers which are superimposed on each other and which differ regarding the orientation  and density of the microfibrils. The arterial subendothelium which is a mixture of amorphous material, microfibrilis, elastin and collagen, with a still unknown orientation of these components, limits the transport of many blood molecules and reinforces the endothelial barrier. A striking difference is observed in the thrombogenicity of these two structures, especially in their reactivity towards platelets. This can be related to the presence of collagen IV in the glomerular basement membrane and of collagen III in the arterial subendothelium: in both cases adhesion of platelets can be observed. The presence of platelet aggregates on the subendothelial surface may also be seen. This is probably due to the presence of collagen III which is known to be able to induce the platelet release reaction and thereby promotes aggregation.


Inhibition of platelet aggregation by cationic protein from polymorphonuclear leukocytes.

The cationic protein isolated from the granules of neutrophilic polymorphonuclear leukocytes inhibited the aggregation of human platelets as induced by collagen, thrombin, ristocetin, aggregated IgG by approximately 50% and anti-C1q by 75% but had no effect on aggregation-induced by ADP.


Galactosyl transferase assay. Application to experimental atherosclerosis.

To establish a repetitive measurement, aortic galactosyl transferase activity has been studied with a specific exogenous acceptor, collagen. Reactions were realized with an acellular biosynthesis mixture containing, collagen (3-4 mg/ml) an aliquot of enzymatic extract preparation (105000 g supernatant (2-3 mg/ml of proteins), UDP (14C) galactose (300 pM/ml). Galactose incorporation into collagen required Mn++ (2.5. 10-3M), incubation temperature of 37 degrees C and pH =7,75. Under such conditions a reproducible assay of aortic collagen galactosyl transferase was possible. After submitting rabbits to a chronic lathyric intoxication and/or to an hypercholesterolimic diet, galactosyl transferase activity was measured in rabbit aortic wall. Enzymatic activity was increased for rabbits under treatment, and the increase was directly proportional to the length of treatment (BAPN associated with cholesterolemic diet).


Biosynthesis of elastin and other matrix-macromolecules in veinous arterial prosthesis.

Homologous saphenous veins where implanted in the arterial circilation (femoral artery) of dogs for various times (1 to 9 months, most data given for 3 months). After this period, the in vitro biosynthesis of intercullar matrix macromolecules was studied by incubating the veins in organ culture conditions in the presence of 14C-lysine for 3 days. A fractional extraction procedure was used to obtain representative macromolecular extracts for the determination of chemical composition (hydroxyproline, hexosamines) and radioactivity. Neosynthesis of elastin was considered as valid criteria for the adaptation of the veinous wall to the new (arterial circulatory) conditions. The chemical composition of the grafted veins was different from that of the nongrafted, controlateral saphenous veins suggesting a molecular remodeling of the grafted veinous wall. Radioactive lysine was incorporated in all macromolecular fractions of grafted and control veins. The specific radioactivity of the extracts obtained from grafted veins was higher than that obtained from the control veins. The only exception was the collagenase extract (containing mainly the polymeric collagen) which had a somewhat lower radioactivity in the grafted vein. The incorporation of lysine in the elastin fraction increased by a factor 10 in the grafted vein as compared to the non-grafted veins. Radioactive desmosine could be demonstrated in the elastin fraction. It appears therefore that the veinous wall could adapt its biosynthetic capacity to the new circulatory conditions by increasing considerably the biosynthesis of elastin. This is accompanied by an increase of the synthesis of other macromolecules and a significant remodeling of the macromolecular structure of the vessel wall.


[Von Willebrand factor activity and thrombophilic states. A review (author's transl)].

After a brief survey of some physiologic properties of Von Willebrand factor activity (VIII r WF) the numerous clinical states with an increase of this activity are described. The authors report their own results concerning sepsis, chronic arteritis of lower limbs, diabetes, toxemia of pregnancy. In most cases the increase of VIII r WF activity appears as a sign of vascular injury and/or the consequence of tissular damage.


Factor V and VIII activation "in vivo" during bleeding. Evidence of thrombin formation at the early stage of hemostasis.

Factor V and VIII activity was measured by a one-stage assay on capillary blood issuing from a skin cut. This activity increases during bleeding. The phenomenon is suppressed by a previous injection of a low dose of heparin. These data give evidence of a very early local formation of thrombin during hemostasis. The study of this activation process in patients with congenital deficiency of blood clotting factor allows to investigate the mechanism of thrombin formation in vivo. This demonstrates the preeminence of the tissue factor pathway and the need of factor VIII for factor X activation in the extrinsic system.


Two simple methods for the quantitative evaluation of platelet adhesion to collagen.

14C serotonine labelled platetel rich plasma (PRP) incubated with fibrillar collagen is either gel filtered through a Sepharose 2B column (Sepharose test) or centrifugated on a 23% Ficoll layer (Ficoll test). In the Sepharose test, the adhesion is quantified by the determination of the per cent yield of free platelets eluted from the column and the release by a quantitative evaluation of the 14C serotonin. In the Ficoll test, the released seroton is measured from the distribution of radioactivity in the different layers (supernatant plasma, platelets adherent to collagen, Ficoll, platelet pellet) of the tube after centrifugation. Both methods are rapid and quite reproducible.


Semantic memory: a bibliography, 1968-1975.

Semantic memory research is briefly described. Then a bibliography of 105 published references in semantic memory, calendar years, 1968-1975, is presented.


Personality and meditation.

The relationship of psychoticism, extraversion-introversion, and neuroticism to the practice of Transcendental Meditation was studied. Male meditators (n = 39) were more introverted and more neurotic than the normal population, whereas the female meditators (n = 27) had a higher mean psychoticism score. Over a 6-mo. period of meditation, the males (n = 24) became less neurotic; the degree to which they did so was related to the frequency of meditation. The implications of these findings are discussed.


Relationship between spatial density and sensory overload, personal space, and systolic and diastolic blood pressure.

Selected effects of high spatial density were investigated with 26 boys, serving as their own controls and undergoing alternate confinements in high and relatively low spatial density. Measured effects were changes in minimal aversion threshold values of white sound, projective measures of desire to alter room size, and systolic and diastolic blood pressure. High spatial density increased systolic and diastolic blood pressure and projective measures of desire to alter room size but did not affect minimal aversion threshold values. Semi-projective measures of personal space requirements were obtained prior to experimental trials. Analysis showed a significant interaction of minimal aversion threshold values with requirements for personal space; the minimal aversion threshold values of subjects requiring large personal space tended to decrease following confinement under high spatial density. The interaction of personal space with projected measures of desire to alter room size was not significant. High spatial density may, then, exert adverse effects on blood pressure and may induce sensory overload conditions in individuals requiring large personal space.


[Isolation and function of platelets. I. Platelet rich plasma. Comparison between 2 methods: gel filtration and albumin density gradient centrifugation. II. A new method using total blood: metrizamide gradient centrifugation].

Human platelets separated from platelet rich plasma (PRP) by two different methods: gel filtraton and centrifugation on albumin gradient have been compared for yield, cellular and protein contamination, ultra-structure, platelet populations, shape change, aggregability and functional preservation. A new method for the separation of platelets from total blood on metrizamide gradients has been established. This technique is rapid and easy; it avoids the initial centrifugation of the PRP where about 30% of the platelets are lost and needs of blood 5 to 10 ml of blood.


Studies on antilipolytic activity of antipyretics. Part I. Influence of sodium salicylate, acetylsalicylic acid, phenazone, aminophenazone, and acetophenidin on lipolysis in fever induced by E. coli pyrogen.

The fever induced by E. coli pyrogen (LPS) is accompanied by a rise of FFA and glycerol level. All tested antipyretics inhibited both thermogenesis of lipolysis produced by LPS. These results suggest that the antipyretic effect of antipyretic drugs is not confined to their action on heat-dissipating mechanisms, but may also be exerted by a depression of lipid metabolism.


Medical versus surgical treatment of stable angina pectoris: progress report of a large scale study.

A large scale, prospective, randomized study of surgical v. medical management of disabling angina pectoris is being conducted as a cooperative study among thirteen Veterans Administration hospitals in the U.S.A. A total of 1015 patients have been entered into the study and follow-up data are currently being evaluated. Patient entry into the study was concluded in December 1974. Patient compliance has been acceptable with only 7% of patients not adhering to their randomization category. Thirty-day operative mortality (1972-1974) in 309 patients was 5-3%. The patient population exhibited a severe degree of coronary disease. There was ECG evidence of prior myocardial infarction in 40%. There were significant obstructive lesions in three major coronary arteries in 51% and significant lesions of the left main coronary artery in 11%. Medical and surgical treatment groups demonstrated no significant differences in objective descriptive characteristics. Mortality in the medical group at 1 year was 8%. Mortality was influenced by several factors including the number of vessels involved, left ventricular function and the presence of left main coronary artery disease. The lowest mortality occurred in patients with single vessel disease and normal LV function who had a 1-year mortality of 3%. Patients with 3-vessel disease and abnormal LV function exhibited a 14% 1-year mortality. Patients with disease of the left main coronary artery and poor LV function had a 1-year mortality of 37%. Analyses of the results of treatment modalities in sub-groups is currently being performed and will be reported in future publications.


Principal aspects of the use of placebos.

After a brief historical review the author discusses the use of placebo in treatment and in the scientific evaluation of the efficacy of psychotropic drugs in double-blind studies. He also mentions its place in the elucidation of the personality structure and psychodynamics of so-called placebo reactors, as well as its importance in recent years for basic medical research.


[Pimozide in non-psychotic elderly patients with behavioural disturbances. An open study (author's transl)].

20 non-psychotic aged (more than 60 years old) in-patients with behavior troubles were treated with pimozide in an open study for more than 12 weeks. Such symptoms deriving from the emotional sphere, mainly the aggressivity, had the best improvement rate. Together with this could also be seen an improvement of the social behavior. The dosage optimum was between 3 and 4 mg p.d., the improvement period seems to reach from the second to the eighth week. Extrapyramidal side-effects were rare, two patients developed, after more than nine months of therapy and long after the study had finished, a tardive hyperkinesia which was reversible after the withdrawal of the substance.


The Comorosan effect: single- and double-blind studies on the urea/urease system.

Characteristic alterations of enzymatic reaction rates by irradiation of the crystalline substrates for fixed periods with an Hg vapor lamp were reported by Comorosan in 1969. Results of single- and double-blind studies reported here support the validity of the key features of this striking and potentially important phenomenon.


[Pharmacoangiography of the pancreas (author's transl)].

The value of pancreatic pharmacoangiography was assessed in 269 visceral angiograms comparing pre- and postpharmacoangiographic series. Angiotensin as a vasoconstrictor was studied in 179 patients, Adrenalin in 50 patients. Experiences with vasodilators based on 40 angiograms carried out with Secretin and Tolazolin were evaluated respectively. After application of Angiotensin, enhanced filling of small arteries in the head of the pancreas was achieved in 75% of patients. Good filling of small arteries of the body of the pancreas rose from 26 to 55%, while angiography with vasoconstrictors seems of low value in the tail of this organ (20% improvement). The diagnostic information of angiography was improved in 4 of 43 carcinomas (10%). Confirmation or exclusion of questionable pathologic alterations are a further, though not measurable advantage of this method. Pharmacoangiography with constrictors is recommended as an alternative method if superselective angiography is not possible. The use of vasodilators for better imaging of the pancreatic parenchyma is of no significant diagnostic value. The use of vasodilators for better presentation of the spleno- and mesenterico-portal axis is advisable because dilatation of the capillary system permits application of larger doses of contrast medium and a higher flowrate. This causes early and more intensive imaging of the veins.


Immunologic aspects of human sarcomas.

Forty-seven patients with mesenchymal sarcoma seen at the Mayo Clinic were tested for humoral and cellular immunologic responsiveness by means of a humoral cytotoxicity test, skin tests for delayed-type hypersensitivity, and lymphocyte transformation with concanavalin A, phytohemagglutinin, and pokeweed. Prospective follow-up of the clinical and laboratory results revealed that patients with low humoral cytotoxic indices, anergic skin test responses, or low lymphocyte transformation responsiveness tended to have the worst short-term clinical outlook.


Resection of malignant bone tumors.

Amputation is an unsatisfactory and depressing measure for the patient as well as for the surgeon. As long as operative removal of tumors in the extermities is indicated, the search for alternative solutions should be the most urgent task for every doctor involved in surgery. Resection would be the most equitable measure as far as demands for radicality and the best possible restoration of function are concerned. In order to warrant the radicality of the operation, meticulous preoperative planning and precise surgical techniques are essential. Sufficient restoration of function must take into consideration all the factors of the individual case, age and profession of the patient playing a large role. After partial resection of longer tubular bones, fitting of an endo-prosthesis has proved to be advantageous. Implants available at present have various drawbacks; studies directed at the elimination of these drawbacks must be set up.


Comparative pathology of bone tumors in animals, with particular emphasis on the dog.

A review of bone tumors in animals, based on cases reported in the literature and on personal examination of cases from several universities in the U.S.A. and Europe, has shown that: 1. The following bone tumors occur in decreasing order of frequency in animals: osteosarcoma, chondrosarcoma, osteochondroma, hemangiosarcoma, fibrosarcoma, and liposarcoma; 2. The dog has by far the highest frequency of bone tumors; and osteosarcomas account for approximately 80 percent of bone tumors in dog, followed by chondrosarcomas and hemangiosarcomas; 3. Bone tumors occur less frequently in the cat than in the dog, and are of essentially the same types and incidence as in the dog; 4. Sheep has a unique propensity for chondrosarcoma and primary bone tumors are rare in other species; 5. Giant cell tumors, reticulum cell sarcomas, and Ewing's sarcomas are exceptional findings in animals.


Biochemical markers in cancer of the breast.

Currently, one could summarize this area by saying that we appear to be in a situation where three relatively nonspecific tests detect the majority of patients with metastatic disease, as well as those post-operative patients who are at high risk of relapse. The critical test of their utility for segregating those at risk for relapse from those who are not at high risk will need to be done in a highly select subgroup, e.g., N- patients. Two of these tests, CEA and hCG, also appear to be useful indicators for predicting the probability of responding to combination chemotherapy in metastatic disease. The development and further testing of potentially more specific markers to replace or add to the current matrix is now in progress, Casein, which is a product of the milk synthesis pathway of breast tissue, represents a potentially more specific test than any of those studied to date. HENDRICK and FRANCHIMONT, 1974, have found elevated levels in 21 of 26, or 81%, of patients with metastatic disease, and 8 of 11, or 73%, of patients preoperatively. The test may also reflect the tumor burden since the proportion of patients with elevated levels dropped to 41-50% postoperatively. Further results with this marker are awaited with interest. Other tests such as ferritin, hydroxyproline, or the development of tumor antigen associated immunospecific assays could increase both the specificity and sensitivity of the tests utilized in this field of investigation. Injecting the use of both single marker tests and matrix approaches into routine clinical use in the postoperative setting now appears to be ready for more critical testing. Their use in diagnostic or screening settings, which is the ultimate goal, also needs to be evaluated. Finally, from the practising clinician's viewpoint the data in this discussion should be considered preliminary. It constitutes a status report. Although there is evidence that CEA and hCG are prognostic in metastatic disease, and that subclinical disease is detectable, larger and more tightly controlled studies will be necessary before their routine clinical use can be recommended in breast cancer patients.


Ketoprofen - clinical efficacy.

Ketoprofen has, over the last few years, undergone extensive clinical trials in many centres throughout the world and these are reviewed. Adequately controlled trials have established its undoubted clinical efficacy as an antirheumatic agent in all the major rheumatic diseases, and have demonstrated therapeutic potency, comparable with other established non-steroidal anti-inflammatory analgesic drugs.


Double-blind cross-over study of ketoprofen and phenylbutazone in patients with chronic osteoarthrosis of the hip.

A double-blind cross-over study of oral ketoprofen capsules 100 mg/day and phenylbutazone capsules 400 mg/day has been carried out in 44 patients with osteoarthrosis of the hip. All patients received each drug for 4 weeks. Eight patients were withdrawn for various reasons during the trial period, leaving 36 assessable patients (20 males and 16 females), whose ages ranged from 31-88 years (mean 66 years). In this series of patients the results suggest that there is little to choose in efficacy between the two drugs at these dosages. All patients showed improvement after 4 weeks' treatment on either drug and of 12 parameters measured only two showed a statistically significant favour to phenylbutazone (intermalleolar straddle and intercondylar distance; both when pain first appeared). Side-effects were reported equally with both medications, even though of necessity the protocol called for the exclusion of patients with previous intolerance or sensitivity to phenylbutazone. The study was instituted during the early evaluation of ketoprofen when the dosage of 100 mg/day was being tried; it is felt that a more significant result may possibly have emerged if the dosage had been the now recommended one of 150 mg/day.


Ketoprofen in the treatment of rheumatoid arthritis.

A double-blind cross-over trial of ketoprofen (200 mg daily) compared with placebo and aspirin (4 g daily) was carried out in 24 patients with rheumatoid arthritis. Ketoprofen was shown to be significantly more effective than placebo in terms of pain relief, degree and duration of morning stiffness, articular index and patients' assessment of improvement. No significant differences were noted between ketoprofen and aspirin, although side-effects were less with ketoprofen.


Double-blind study of ketoprofen and phenylbutazone in ankylosing spondylitis.

A series of 26 patients with ankylosing spondylitis have been included in a double-blind cross-over comparison of ketoprofen 200 mg daily and phenylbutazone 300 mg daily, each drug being given for 4 weeks, with paracetamol as a 'rescue drug'. Six patients withdrew from the study: three on phenylbutazone and one on ketoprofen--two defaulted treatment. Twenty patients (17 males and 3 females) were eligible for assessment. Their ages ranged between 20 and 59 years (mean 40.2 years). Patient characteristics were comparable in both groups. The results of this series indicate no suggestion of statistically significant differences in any of the nine movements measured. The only significant differences, both in favour of phenylbutazone, were in the paracetamol consumption in the second treatment period in the group receiving ketoprofen first (0.01 greater than P greater than 0.005) and in the patients' preference for phenylbutazone (0.005 greater than P greater than 0.001). Adverse reactions were comparable in both groups.


Clinical evaluation of keto profen (Orudis) in rheumatoid arthritis: results of a multi-centre double-blind trial against phenylbutazone.

In a double-blind cross-over trial in rheumatoid arthritis, no difference was found between ketoprofen and phenylbutazone with regard to clinical efficacy at the dosages used. There was a higher incidence of side-effects reported during treatment with phenylbutazone.


A double-blind trial of ketoprofen in the treatment of osteoarthritis of the hip.

In a double-blind study of ketoprofen and placebo in the treatment of osteoarthritis of the hip, ketoprofen was shown to be significantly more effective. Analysis of results was made using the sequential technique. Major intolerance occurred in two cases and minor intolerance in five cases. Newly diagnosed cases were treated more easily with ketoprofen than chronic cases treated for several months with other drugs which had proved ineffective. There were no changes in biological parameters. Age and sex did not affect the result. The further study of ketoprofen in large open trials appears to be indicated.


Clinical evaluation of ketoprofen (Orudis) in rheumatoid arthritis: a double-blind cross-over comparison with indomethacin.

In a double-blind cross-over trial, ketoprofen was found to be as effective as indomethacin in the treatment of rheumatoid arthritis. Side-effects, particularly headache and vertigo, were less common in patients receiving ketoprofen.


Double-blind trial of ketoprofen compared with placebo in osteoarthrosis of the hip.

In this study, patients suffering from osteoarthrosis of the hip received two successive periods of treatment, A and B, each of two weeks, with either ketoprofen or a placebo. The daily dosage of ketoprofen was 150 mg in three divided doses of 50 mg. In each case the findings necessary for diagnosis and for assessment of progress during the treatment were recorded. At the end of the trial, the patient's preference for one or other of the two periods of treatment was used as the criterion. The results were analysed by the sequential method. The therapeutic effects assessed by the various criteria included in the protocol (patient's subjective assessment, pain at rest, duration of morning stiffness, distance walked without pain, range of movements, etc.) were also recorded. Nine cases were sufficient to provide a statistically significant result in favour of ketoprofen. In eight cases the preference was definitely in favour of ketoprofen. In a single case there was no preference for one or other of the two periods of treatment (failure of both ketoprofen and placebo). Tolerance to ketoprofen was excellent.


Results of the rectal administration of ketoprofen in fifty-two patients.

The results of a clinical study of ketoprofen suppositories suggest that the drug is effective when given in this way. It is suggested that, in the treatment of inflammatory rheumatic disorders, advantage should be taken of the longer duration of action of ketoprofen suppositories to prescribe their administration early in the morning and before retiring. Local intolerance occurred in only 4 of 52 patients.


A clinical study of ketoprofen suppositories.

Ketoprofen administered via the rectal route seemed to be valuable when given at night to patients with various rheumatic syndromes and may be particularly useful for patients who show gastric intolerance of the capsules. Anal intolerance was noted in 12% of the patients.


Clinical study of ketoprofen administered rectally in rheumatology.

The effects of ketoprofen suppositories were studied in an open trial in 50 patients with various rheumatic disorders. A beneficial effect was obtained in many patients and this lasted for about 8 hours after administration. Mild local irritation was the commonest side-effect but seldom led to withdrawal of treatment.


Ketoprofen suppositories in rheumatological practice.

Preliminary results are reported on 30 patients treated with suppositories of ketoprofen at a dosage of two daily (200 mg). Local side-effects, rectal burning and difficulty in retaining the suppository, were recorded in seven patients and caused treatment to be stopped from the first day, and in five others local (burning) or systemic (gastric pain) side-effects caused treatment to be abandoned at a later period. Interest in the suppository presentation may lie in its more prolonged action and this presentation, when given at night, may improve the over-all results obtained with ketoprofen capsules in inflammatory rheumatism.


A study of ketoprofen in rheumatoid arthritis.

Experience with ketoprofen, using the oral and rectal routes, was reviewed. The value of ketoprofen in rheumatoid arthritis was confirmed and the drug was usually well tolerated.


A controlled trial of ketoprofen (administered rectally) in arthritis of the hip and knee.

A double-blind therapeutic trial carried out by the sequential method has demonstrated clearly a significant difference in efficacy between ketoprofen (given rectally at a dosage of 200 mg daily) and a placebo in a group of patients suffering from osteoarthritis of the hip or knee. The interest in this type of study lies in the fact that it will often given the answer to a simple question using a relatively small number of patients.


Clinical trial of the injectable form of ketoprofen.

Although the number of synthetic anti-inflammatory agents is ever increasing, it is apparent that most of these drugs have been administered orally or rectally. Because it can be given parenterally, ketoprofen is of additional interest. Furthermore, tolerance of the product by the intramuscular route is adequate even in patients likely to suffer side-effects when the drug is given orally.


Clinically therapeutic study of intramuscular ketoprofen in rheumatic diseases.

We have studied the effect of ketoprofen (19583 R.P.) given intramuscularly in some painful rheumatic conditions, notably root pains of disc and spinal origin and inflammatory conditions, and it has proved to be an effective analgesic and anti-inflammatory product, and in our series of 30 patients was very well tolerated.


Comparative study of intravenous ketoprofen versus aspirin.

In order to assess the analgesic effect of an injectable preparation of ketoprofen, a double-blind comparative study was conducted against a soluble form of acetylsalicylic acid of known efficacy, widely used in the clinic. Only one indication was considered for inclusion in the trial, namely discogenic sciatica. We found no significant statistical difference between these two products, considering the criteria used. The time of onset of effect was, on average, 30 minutes and the mean duration of activity was 4 hours. The two products were well tolerated.


[Diagnostic and therapeutic problems in dystonia musculorum deformans].

Taking as starging point three patients who unsuccessfully underwent both medical treatment and different neurosurgical interventions, the Authors discuss the problem of the dystonia musculorum deformans nosographic arrangement and its present therapeutical possibilities. They believe that the dystonia musculorum deformans must be considered, rather than as a syndrome, as a real disease, to be distinguished in particular from the various infantile dystonic syndromes and from spasmodic torticollis. In fact in these last morbid forms, at least from a neurosurgical standpoint, the therapeutical possiblities are not so uncertain as those of the dystonia musculorum deformans.


Estimation of lymphocyte subpopulations in the peripheral blood of patients with sarcoidosis.

Lymphocyte subpopulations in the peripheral blood of 36 sarcoidosis patients and 36 age- and sex-matched controls were determined, using the sheep erythrocyte rosette (SRBC-R) technique and immunofluorescence staining with whole rabbit anti-human immunoglobulin antisera. In the patient group the total number of lymphocytes, SRBC-R lymphocytes, and IgM-positive lymphocytes was reduced in comparison to in the control group. In the patient group no correlation could be found between any of the lymphocyte subpopulations and disease stage, cutaneous involvement of the disease, or cutaneous reactivity to purified protein derivative and dinitrochlorobenzene.