Series on pharmacology in practice: 1. Drugs that alter mood. I. Tricyclic agents and monoamine oxidase inhibitors. In the last 20 years, the treatment of mood disorders has advanced immeasurably. We now have relatively safe and effective agents for the treatment of depression and mania. This review discusses two types of agents that elevate mood--tricyclic antidepressants and monoamine oxidase inhibitors--including the indications for their use and their modes of action, pharmacokinetics, side effects, and drug interactions.


Health status index: category rating versus magnitude estimation for measuring levels of well-being. Levels of Well-Being are social preferences, or weights that members of society associate with time-specific states of function. A Weighted Life Expectancy, which can be used to measure program outputs, is created by summing the levels across diverse cases and multiplying them by probable transitions (prognoses) among the states and levels. This operation requires however, that the Levels of Well-Being be measured on underlying metric scale. The present analysis compares preference measurements from a simple category rating procedure with those obtained using the more complex and difficult magnitude estimation method which has been claimed to yield ratio level measures. In a randomly counterbalanced design, 65 college students rated 30 case descriptions representing the range of the Well-Being continuum. The results exhibit the classical logarithmic relation observed for a prothetic continua. When transformed to a meaningful 0-1 unit scale, however, the magnitude responses are compressed at the lower end of the scale near death. Such results are inconsistent not only with category rating, but also with intuitive notions of the relative importance of the function states, with the results of rating procedures that simulate social choice, and with evidence that confirms the interval properties of the category ratings themselves. Furthermore, the ease of administration of category rating means that multiple attributes of cases can be considered jointly, avoiding the need to aggregate scale values for different attributes by arbitrary rules. In sum, magnitude estimation is inappropriate as a measurement method for a Health Status Index and is probably also inappropriate for other measures of utility and social choice.


[Determinación of islet-cell antibodies in different groups of diabetics (author's transl)]. The discovery of islet-cell antibodies (ICA) has served to reinforce the theory that the pathogenesis of diabetes is immunologic. These antibodies have been found in the serum of diabetics who also had other autoimmune diseases and in that of most insulin dependent diabetics. ICA were detected by an indirect immunofluorescent technique using human pancreas tissue as a substrate. In this study two human islet cell adenomas were employed; their cell composition was analyzed by means of this technique with antihormone antibodies marked with fluorescein. Serum from 74 diabetic patients and 45 normal controls were studied using this substrate. Twenty-three of the 56 insulin-dependent diabetics were in the first clinical stage of the disease; 18 of them (78%) were positive for ICA. The other 33 insulin-dependent diabetics had the disease for over 3 years; six of these (36%) were positive. Anticellular antibodies were detected also in two of the 18 patients who were not insulin-dependent (5%). There was only one case of positivity among the controls (2%). These figures are similar to those of previous studies. It has been shown that the incidence of islet-cell antibodies drops rapidly after the fifth week of the disease; it depends primarily on the stage of development and has little to do with the age of the patient at the onset of the disease. Knowledge about these antibodies makes it easier to classify diabetics and to arrive at a prognosis regarding possible later insulin-dependence. These antibodies may be a consequence or even the cause of the destruction of the pancreatic islets provoked by a viral infection and perhaps determined by a predisposing genetic factor.


[A study of the alveolar-arterial oxygen gradients and intrapulmonary shunt under the action of NAB-365, a new exogenous catecholamine of selective beta-2 action (author's transl)]. Thirty patients were studied using NAB-365 (clenbuterol), an exogenous catecholamine with selective beta-2 action to determine various parameters of pulmonary physiopathology. They included: alveolar-arterial gradients in partial oxygen pressures, coefficient of respiratory inefficacy, alveolar ventilation, and intrapulmonary venoarterial short circuits. By studying these parameters before and after clinical administration of clenbuterol we were able to demonstrate a decrease in the alveolar-arterial oxygen gradients (p less than 0.001), decrease in the respiratory inefficacy coefficient (p less than 0.001), increase in alveolar ventilation (p less than 0.001), and a shunt decrease (p less than 0.0025). The drug improves diffusion and oxygenation without affecting the adrenergic cardiac receptors.


The rhinologist and the management of pituitary disease. Since the early days of pituitary surgery, a variety of transnasal approaches have been used to gain access to the sella turcica. Each of these approaches requires crossing the sphenoid sinus, hence the transsphenoidalnoidal designation of these methods. Since the growth and refinement of microsurgery as a distinct surgical discipline, there has been a coincidental maturation of transsphenoidal microsurgical techniques for the management of a variety of pituitary disorders. The present paper reviews the historical events leading to the current methods of transphenoidal pituitary surgery. Detailed descriptions of each method are given, with the advantages and disadvantages of each. The author's method is described and illustrated in a step-by-step manner, and the criteria which determine operability by transsphenoidal versus transfrontal craniotomy are discussed. The advantages which transsphenoidal techniques offer over transfrontal methods are emphasized. The paper deals with pituitary disorders on a clinical and pathological basis. The work-up of pituitary lesions is discussed, with a detailed presentation of the endocrine evaluation. Further emphasis is given to the value of tomography of the sphenosellar complex in planning operative approaches for removal of pituitary lesions, particularly when the lesion is a so-called "microadenoma," producing no obvious expansion nor erosion of the cellar floor. The author's experience in the management of 125 lesions of the pituitary or associated sella turcica is presented and complications are discussed. Because of the relatively brief period of follow-up, no conclusive evidence can be offered regarding cures in the author's series. However, on the basis of reported experience by others working in the same area, one may expect acceptable long-term improvement or cures in selected pituitary lesions operated on by transshpenoidal methods. The team management of pituitary disorders is stressed. In particular, emphasis is given to the contributions which the current day otorhinolaryngologist can make, because of his knowledge and experience in rhinological and microsurgical techniques. There is no other surgical specialty so uniquely qualified to provide operative treatment of pituitary lesions using transsphenoidal surgical pathways.


Flap reconstruction in patients with head and neck cancer: techniques and results. Patients who are treated by surgery, radiotherapy, or any combination thereof may suffer from complications in wound healing or tumor recurrence which necessitate reconstruction of repair utilizing tissue brought in from other parts of the body. The majority of patients, who required flap reconstruction for primary repair or for a complication of their surgery, had their primary lesion in the supraglottic larynx or inferior hypopharynx. More than two-thirds of the flaps elevated were for fistula repair. Five year survival in this group of patients was lower than in those patients with uncomplicated postoperative courses, although the quality of life for the survivors in both groups was not significantly different.


[Automation of the determination of cardiac output by means of 4-electrode rheocardiography]. A method of cardiac output determination is described, based on automatic processing of the differential rheogram and arterial pressure traces, by a computer algorithm. All variable parameters of the rated formula are defined automatically. The method provides highly precise and rapid definition of the cardiac output.


[Laser correlation spectrometer for immunological and virological analyses]. A correlative laser spectrometer is recommended to record the immunologic reactions and differentiate the virus particles. The correlator used in this spectrometer is of the three-bit type; it measures the base line and allows preliminary signal processing. The correlative laser spectrometer is designed for application in clinical diagnostic laboratories.


[Automated system of medical examination data processing]. To solve some problems concerning practical and experimental medicine, an automatic system was elaborated for processing the medical examination data. The core of the system is the NAIRY-2 computer. All the problems to be solved were divided into four groups, according to the software management: those, concerned with archival storage and acquisition of data about the patients' state; on formation of visual (graphic) presentation of the data contained in perforated documents; on diagnosis and prognoses, according to clinical and physiological data through the use of an applied program package; and those to be programmed by the user, in accordance with standard and additional software of the NAIRI-2.


[Cardiosynchronized means of treatment and diagnosis]. Diverse ways of a cardiophased action on the cardio-vascular system for therapeutic and diagnostic purposes are assessed. Requirements for the basic subunits of the assisted circulation apparatus to be met, the possibilities open for developing automatic cardiosynchronization systems, and also features peculiar to the synchronization mode in effecting coronarography and conditions necessary for implementation of the method for elective introduction of liquid agents into the supravalvular space of the aorta are discussed.


Plasmid replication functions. II. Cloning analysis of the repA replication region of antibiotic resistance plasmid R6-5. R6-5 is a low copy number, conjugative, FII incompatibility group plasmid that has a molecular length of 102 kb and that specifies resistance against several antibiotics (chloramphenicol, fusidic acid, kanamycin, streptomycin and sulphonamide) and mercury salts. By means of in vitro cloning procedures, mini plasmids have been generated that contain a DNA segment from the essential region of R6-5 that is only 2.6 kb in length. This DNA segment, which consists of two PstI fragments that are adjacent in the parent plasmid, carries all genes and sequences required for the regulated replication and incompatibility properties of R6-5, including its origin of replication, OriV, an essential function that has been designated RepA, and the copy control function, Cop. Three different polypeptides, having monomer molecular weights of 23,000, 10,000 and 9,500 daltons, are synthesized in detectable quantities by minicells carrying pBR322 hybrid plasmids that contain DNA segments from the R6-5 essential region. A spontaneous deletion derivative of a pBR322 hybrid plasmid that carries the R6-5 origin of replication was isolated. Heteroduplex analysis of this derivative plasmid indicates that the deleted DNA segment carries the R6-5 replication origin and that its termini consist of short inverted repeat sequences.


Inserted sequence in the mitochondrial 23S ribosomal RNA gene of the yeast Saccharomyces cerevisiae. The sequence organization of the yeast mit-DNA region carrying the large ribosomal RNA gene and the polar locus omega was examined. Hybridization studies using rho- deletion mutants and electron microscopy of the heteroduplexes formed between 23S rRNA and the appropriate restriction fragments, lead to the conclusion that the 23S rRNA1 gene of the omega+ strains is split by an insertion sequence of 1,000-1,100 bp. In contrast, no detactable insertion was found in the 23S rRNA gene of the omega- strains. The size and the location of the insert found in the 23S rRNA gene of the omega+ strains appear to be identical to those of the sequence delta which had previously been found to characterize the difference (at the omega locus) between the mitDNA of the wild type strains carrying the omega+ or omega- alleles (Jacq et al., 1977).


Plasmid replication functions. III. Origin and direction of replication of a "mini" plasmid derived from R6-5. Replicating DNA molecules of the mini R6-5 plasmid, pKTO71, were purified by equilibrium centrifugation in two successive ethidium bromide-caesium chloride gradients, converted to linear forms by cleavage with either HindIII or BglII restriction endonuclease, and examined in the electron microscope. Determination of the replication fork positions in 65 replicating molecules demonstrated that replication is initiated at a unique location on the plasmid and that it proceeds uni-directionally from this site. The direction of replication is such that the origin-proximal BglII cleavage site is replicated late or, in the case of the parent R6-5 plasmid, is such that the R-determinant region of the molecule is replicated early. The origin of replication, located by these experiments at R6-5 coordinate 98.6 kb, is clearly distinct from that of the R6-5 incompatibility determinant which has been shown to be located on an adjacent PstI-generated DNA fragment whose termini have R6-5 coordinates 96.8 and 97.9 kb. This result indicates that the incompatibility function is not an origin DNA sequence.


Introduction of an active enzyme into permeable cells of Escherichia coli: acquisition of ultraviolet light resistance by uvr mutants on introduction of T4 endonuclease V. Plasmolysed cells of Escherichia coli N212 (uvr A recA) acquired ultraviolet resistance when the cells were exposed to high concentrations of T4 endonuclease V. With increasing concentrations of T4 enzyme, survivals of plasmolysed cells after ultraviolet irradiation increased while colony-forming ability of unirradiated plasmolysed cells was not significantly affected by the enzyme treatment. Under appropriate conditions more than 200 fold increase in survivals was observed. When plasmolysed cells were treated with a pre-heated enzyme preparation or enzyme fractions derived from T4v1 (endonuclease V-deficient mutant)-infected cells, only little or no reactivation took place. Permeabilization of cells prior to the enzyme treatment was essential for the effective reactivation. Treatment of intact cells with the T4 enzyme did not cause any reactivation. Cells treated with 20 mM EGTA or 50 mM CaCl2 in cold were reactivated to certain extents by the enzyme, but the extents of the reactivation were far less compared to those of plasmolysed cells. Plasmolysed cells of strains carrying a mutation in one of uvrA, uvrB and uvrC genes were reactivated by introduction of T4 endonuclease V, as was the uvrA recA double mutant. UvrD mutants were also reactivated, but rather slightly. However, wild type strain as well as strains having a mutation in recA or polA gene were not reactivated. From these results it was suggested that T4 endonuclease V, taken up into permeable cells, can function in vivo to replace defective functions, which are controlled by the uvr genes. The conditions established in the present study may be used for introduction of other proteins into viable bacterial cells.


Studies of viable T4 bacteriophage containing cytosine-substituted DNA (T4dC phage). II. Cleavage of T4dC DNA by endonuclease SalI and bam HI. Digestion of non-glucosylated and cytosine-substituted T4 phage (T4dC) DNA with SalI restriction endonuclease showed that the DNA had nine SalI-sensitive sites. There were eight SalI sites in DNA from a strain which had a deletion in the rII-denB-ndd region. The comparison of two digestion patterns indicated that one of the SalI-sensitive sites was present in the deleted region and that the SalI-F fragments (8.4 x 10(6) daltons) was located adjacent to the SalI-C or SalI-D fragments (15.5 x 10(6) daltons) on the T4 chromosome. The DNA gave no detectable cleavage product when digested with BamHI endonuclease alone, while, when digested successively with BamHI and SalI, the DNA yielded two new digestion products in place of one fragment formed by SalI alone. The BamHI-sensitive site was in the SalI-A fragment (25.2 x 10(6) daltons). The usefulness of this information for making cleavage maps of T4 phage chromosome is discussed.


Degradation of Escherichia coli DNA: evidence for limitation in vivo by protein X, the recA gene product. DNA is more extensively degraded after it is damaged in recA mutants of E. coli than in wild type cells. All data presented here are consistent with the recA gene product, protein X, being an inhibitor of nalidixic acid induced degradation of the bulk DNA (but not of newly replicated DNA). Production of protein X also is correlated with appearance of various "S.O.S." repair functions. Evidence was obtained by comparing the rates of protein X synthesis and solubilization of uniformly-labeled DNA in intact cells, incubated in the presence of nalidixic acid. A set of mutants at the lexA locus produced protein X at different rates and degraded their DNA at rates which were inversely correlated to their rates of protein X production. A low concentration of rifampicin quite specifically inhibited protein X production by wild type E. coli, and allowed more rapid DNA degradation. After the DNA was damaged by the incubation of cells in the presence of nalidixic acid, cells preloaded with protein X degraded their DNA more slowly. We propose that protein X could protect DNA against degradation by binding to single-stranded regions, thereby inhibiting nuclease action.


The use of sym-triazine trichloride in RNA-protein cross-linking studies with Escherichia coli ribosomal subunits. The reagent sym-triazine trichloride is used as a bifunctional reagent to generate RNA-protein cross-links within intact ribosomal subunits from E. coli. The reaction takes place in a stepwise manner, involving substitution of one chlorine atom at 12 degrees and pH 8, and substitution of the second at 40 degrees and pH 6. The cross-linked proteins are analysed by two-dimensional electrophoresis, and the existence of a stable cross-linkage is demonstrated by isolating protein-oligonucleotide complexes from 32P-labelled subunits. The proteins cross-linked are S3 and S4 in the 30S subunit, and L2 in the large subunit, together with smaller amounts of other proteins. The reagent should prove useful in topographical studies of the E. coli ribosome as it is a rigid molecule and generates very short cross-links.


Inhibition of growth by imidazol(on)e propionic acid: evidence in vivo for coordination of histidine catabolism with the catabolism of other amino acids. Imidazole propionic acid (ipa), a gratuitous inducer of the histidine-utilization (hut) system in Salmonella typhimurium, inhibits the organism's growth on succinate minimal medium. Induction of the hut system is necessary, but not sufficient, to cause inhibition. A study of the ability of single amino acids to relieve ipa-restricted growth suggests that insufficient glutamate is the cause of slow growth. The inhibition of growth by imidazolone propionic acid (iopa), an intermediate in the catabolism of histidine to glutamate, is similar to that by ipa. Studies using 2, 3, 5-triphenyl tetrazolium chloride plates to examine amino acid catabolism suggest that accumulation of ipa or iopa leads to inactivation of aspartate amino-transferase (AAT). This interpretation is supported by studies of an Escherichia coli mutant lacking AAT. The mutant grows poorly on succinate minimal medium, and the poor growth is relieved by the same amino acids that relieve ipa- and iopa-restricted growth. These and other findings are discussed in terms of coordination of the histidine-utilization system with enzymatic activities involved in the catabolism of other amino acids.


X-ray sensitivity of Escherichia coli lysogenic for bacteriophage P2. Strains of Escherichia coli C or K lysogenic for the non-inducible phage P2 show a lower survival following X-ray irradiation as compared to nonlysogenic strains. This difference in X-ray sensitivity is not accompanied by a significant difference in X-ray induced mutability. The capacity of X-irradiated P2 lysogens to multiply any of a number of unirradiated infecting phages is severely impaired. These effects of X-ray treatment can be most simply explained as a consequence of the fact that protein and RNA syntheses are strongly inhibited in P2 lysogens after X-irradiation. All the above events specifically occurring in X-rayed P2 lysogens are dependent on the P2 gene old.


Regulation of RNA synthesis in yeast. III. Synthesis during the cell cycle. Centrifugal elutriation was used to separate cells in different stages of the cell cycle from a culture of Saccharomyces cerevisiae in balanced exponential growth. The rate of DNA and RNA synthesis was determined using a pulse--long-term label technique that is capable of distinguishing between exponential, linear, and periodic variations in the rate of synthesis through the cell cycle. It was found that while the rate of DNA synthesis varies periodically through the cell cycle, the rate of synthesis of mRNA, rRNA, and tRNA increases exponentially through the cell cycle. The implications of these findings for the control of RNA synthesis are discussed.


Genetics of ribosomal protein methylation in Escherichia coli. III. Map position of two genes, prmA and prmB, governing methylation of proteins L11 and L3. Two genes governing ribosomal protein methylation have been located on the map of Escherichia coli by conjugation and transduction crosses between wild-type and prm (protein methylation) mutants. The Prm phenotype of recombinants was determined by an in vitro assay of methylgroups incorporation into protein. Gene prmA, governing methylation of protein L11 is situated at minute 71 on the map and is cotransduced with aroE (30%) and with rpsL (5%). Gene prmB, governing methylation of protein L3 is at minute 50, very close to aroC (98.5% co-transduction). A cold-sensitive phenotype was found associated with mutation prmB and was used to score a large number of recombinants in a three factor cross. The results of this cross suggest the order aroC -prmB - purF. The striking symmetrical clustering of aro, prm and rim (ribosome maturation) genes is discussed.


Analysis of rpsD mutations in Escherichia coli. I. Comparison of mutants with various alterations in ribosomal protein S4. Streptomycin-independent revertants were selected from streptomycin-dependent mutants. Twenty-five out of 150 such revertants were temperature sensitive. Ribosomal proteins from 18 temperature-sensitive and 10 temperature-insensitive revertants were analysed by SDS-polyacrylamide gel electrophoresis. Seventeen of the former but none of the latter category showed an alteration of protein S4. The mutated rpsD allele of 6 temperature-sensitive revertants was transduced into a rpsL+ strain. In all cases an increased suppressibility of T4 amber phages was observed. Such suppressibility was not observed in the original rpsD, rpsL strains. All 18 temperature-sensitive mutants were disturbed in the processing of 17s to 16s RNA at non-permissive temperature and the accumulated 17s RNA was degraded. Temperature-insensitive rpsD revertants could be isolated, which had gained a second alteration in S4. Such revertants, which had lost the temperature-sensitive property, were also unable to suppress growth of T4 amber phages. It is concluded that temperature-sensitive growth, inability to process 17s RNA and to assemble 30S ribosomes at non-permissive temperature as well as increased translational ambiguity are highly correlated properties in rpsD mutants.


Analysis of rpsD mutations in Escherichia coli. II. Physiology of some representative mutants. The effects of ribosomal ambiguity mutations (ram A-) on the assembly of ribosomal 30S subunits in Escherichia coli were studied in some representative mutant strains. It was found that the inability of these strains to produce active 30S subunits at nonpermissive temperatures is correlated with a halt in the accumulation of protein S4. It is demonstrated that 30S-precursor particles lacking this protein accumulate and break down at nonpermissive temperatures and that most of the 30S proteins as well as the 17S RNA constituting these particles are similarly unstable. These findings are discussed and related to the finding that merodiploid strains containing genes for both mutant and wild type protein S4 do not accumulate the mutant form of the protein. Experiments indicating that ribosomal precursor particles are associated with polysomes are presented. The implications of these findings are discussed and it is suggested that the assembly of ribosomes is tightly coupled to the synthesis of ribosomal proteins.


Analysis of rpsD mutations in Escherichia coli. III. Effects of rpsD mutations on expression of some ribosomal protein genes. Relative rates of production and steady state levels of ribosomal proteins were determined in a temperature sensitive rpsD (S4) mutant of Escherichia coli. Some proteins (S4, S12, S13) were overproduced in the mutant at permissive temperature but steady state levels of all examined ribosomal proteins were normal. In a rpsD+/rpsD+ homodiploid strain the relative rates of production of ribosomal proteins were not affected by the increased gene dose. In a rpsD+/rpsD heterodiploid strain only wild type, but not mutant S4, was found. In such a strain S4, S7, S12 and probably S13 is overproduced. It is implied that S4 is involved in the regulation of expression of proximal genes of the two transcriptional units including the genes coding for S4 itself and S12, respectively. A degradation system for ribosomal proteins, which is rapid enough to be of regulatory significance, is demonstrated.


W-reactivation of phage lambda in recF, recL, uvrA, and uvrB mutants of E. coli K-12. W-reactivation is reduced by recF143 and recF144 mutations and is undetectable if a second mutation at either the uvrA or uvrB locus is combined with recF143. The uvrA and uvrB mutations alone block W-reactivation partially. A recL152 mutation also partially blocks W-reactivation by itself. In combination with a uvrB5 mutation, recL125 blocks W-reactivation completely but in combination with recF143, significant residual W-reactivation ability remains. We suggest that the phenomenon of W-reactivation is the result of at least two modes or pathways. The observation that recF143 uvrB5 and recF143 uvrA6 strains permit normal levels of mutagenesis (Kato et al., 1977) but completely block all W-reactivation leads us to suggest further that the mechanism(s) of W-reactivation is at least partly different from that of UV mutagenesis.


Deletion mapping of the ilvGOEDAC genes of Escherichia coli K-12. A set of lambdadilv phage has been examined that carry overlapping segments of isoleucine-valine structural and regulatory genes derived from the ilv cluster at 83 min on the Escherichia coli K-12 chromosome. The ilv genes present in these phage, and their order, have been determined by transduction of auxotrophs, escape synthesis, and deletion mapping. The order of ilv genes in the phage, and hence the order in the host chromosome, was found to be ilvG-ilvO-ilvEDA-ilvC. Lysogens containing lambdadilv phage were constructed for dominance analysis of regulatory mutations in the ilvO and ilvA genes. The ilvO671 allele is cis-dominant to ilvO+, while the ilvA538 allele is trans-recessive to ilvA+. Thus, the ilvO gene, that is identified by cis-dominant regulatory mutations that result in increased ilvG and ilvEDA expression, is situated between and may be contiguous with ilvG and ilvEDA.


Mutations affecting the formation of acetohydroxy acid synthase II in Escherichia coli K-12. Genetic mapping experiments have established that two recently isolated valine-resistant mutants of the K-12 strain of Escherichia coli have lesions lying between ilvE and rbs. These lesions allowed expression of the ilvG gene, specifying the valine-insensitive acetohydroxy acid synthase (synthase II) and an increased expression of the ilvEDA operon. In this respect, they resembled an earlier described ilvO lesion that was reported to lie between ilvA and ilvC. All three lesions were cis-dominant in cis-trans tests. Reexamination of the earlier studied ilvO lesion revealed that it, too, lies between ilvE and rbs. Valine-sensitive derivatives with lesions presumed to be in ilvG were selected from each of the valine-resistant strains. In two of the valine-resistant strains, the ilvG mutations were on the rbs side of ilvO, indicating a gene order rbs-ilvG-ilvO-ilvE-ilvD-ilvA-ilvC. In one of the recently isolated valine-resistant stocks, however, the apparent ilvG mutation was found to be between ilvE and the aline resistance marker. This finding suggests that either ilvO and ilvG mutations are interspersed or there is another locus, ilvR, that behaves phenotypically like ilvO and which lies between ilvG and rbs.


Mutants of Escherichia coli defective in the degradation of guanosine 5'-triphosphate, 3'-diphosphate (pppGpp). A new class of mutants of E. coli exhibiting altered metabolism of ppGpp and pppGpp has been isolated, and mapped at a locus designated gpp, near min 83 on the genetic map. These mutants accumulate elevated levels of pppGpp during amino acid starvation or carbon source downshift, and exhibit a reduced rate of pppGpp degradation in vivo. The in vitro evidence suggests that the gpp mutants are defective in a 5'-nucleotidase, which specifically hydrolyzes pppGpp to ppGpp. Certain combinations of gpp and spoT mutations are inviable. A gpp spoT double mutant, constructed by employing a leaky spoT mutation, was found to have a slower rate of pppGpp degradation than the gpp mutant alone. This result indicates that spoT also participates in pppGpp degradation. The inviability of certain gpp spoT combinations is attributed to the inability of the double mutants to degrade pppGpp. This is supported by the observation that selection for increased growth rate on the double mutant results in the recovery of relA mutations. Various effects of the gpp mutation upon the pppGpp and ppGpp pools provide additional support for a scheme in which pppGpp is the major precursor of ppGpp.


Genetic organization of the E. coli chromosome around the structural gene for initiation factor IF3 (infC). A set of lambda transducing phages carrying varying lengths of the E. coli chromosome around the structural gene for initiation factor IF3 (infC) was derived from lambda p2 which is known to carry, besides infC, the structural genes for the alpha subunit of phenylalanyl-tRNA synthetase (pheS), the beta subunit of phenylalanyl-tRNA synthetase (pheT) and the structural gene for threonyl-tRNA synthetase (thrS). The E. coli coding content of these derived phages was analysed by genetic complementation of a set of mutants and by SDS-polyacrylamide gel analysis of the proteins synthesized in UV irradiated cells infected with these phages. The segregation pattern of the different genes among these derived phages indicates that the order of the genes is pheT - pheS - "P12" - (infC, thrS) where infC is probably between "P12" and thrS. "P12" is the structural gene of a 12,000 molecular weight unidentified protein.


Investigations of the F conjugation gene traI:traI mutants and lambdatraI transducing phages. A series of traI point and deletion mutants of Flac, and a traM mutant, were characterised. Complementation tests with an amber Flac traI mutant confirmed their genotypes, and in addition all the traI mutants, but not the traM mutant, were complemented by pRS31 (PSC101 traDI) and EDlambda109 (lambdatraI). Judging from the efficiencies of plating of F-specific phages, none of the mutations affected pilus formation. The traI products of F and of the F-like plasmid R1 were interchangeable with each other but not with that of R100, while the traM product of F could not be replaced by those of R1 or of R100. Neither traI nor traM were needed for conjugal transfer of ColE1. Three lambda transducing phages carrying traI were isolated by in vivo or in vitro techniques, and characterised by genetic complementation tests, by analysis of the fragments produced by restriction endonucleases, and by measurement of heteroduplex molecules. The genetic structures together with the sizes and F coordinates, of the transfer regions carried by the phages were thereby determined. Comparison of the proteins synthesised in UV-irradiated cells by one of the lambdatraI phages with those made by a derivative carrying an amber traI mutation, allowed the traI product to be identified as a protein of molecular weight 174,000. In addition, the molecular weights of the traD (84,000), traS (18,000), and traT (25,000) products made by the lambdatraSTD1 phage EDlambda107 were measured. The possible roles of the traI and traM products in conjugation are discussed.


Analysis of lincomycin resistance mutations in Escherichia coli. High level lincomycin resistant strains of Escherichia coli were isolated and screened for altered ribosomal proteins and functions. Amongst 58 strains investigated by electrophoresis one had an altered ribosomal protein S7, another one a mutated L14 and two showed altered L15 proteins. A correlation between these alterations and lincomycin resistant growth could not be demonstrated by genetic analysis for any of the mutants. In vitro, however, extracts from the two L15 mutants were less sensitive to inhibition by the drug. A gene locus (linR) responsible for the lincomycin resistance phenotype was mapped at min 30 of the Escherichia coli chromosome near tyrR; it seems to be identical to the previously described linB locus (Apirion, 1967); however, in contrast to these reports it does not seem to alter any ribosomal function.


Individual messenger RNA half lives in Saccharomyces cerevisiae. We have measured the decay half-life of functional messenger RNA (mRNA) for some thirty different proteins in the yeast Saccharomyces cerevisiae. Production of newly synthesized mRNA was halted by raising the temperature of a culture of a temperature-sensitive mutant, ts 136. Aliquots of this culture were pulsed-labelled with [35S]-methionine at various times after the temperature shift and the radioactive proteins separated on the two-dimensional gel electrophoresis system of O'Farrell. We find a range in the decay half lives of individual mRNA species which varies from 3.5 min to greater than 70 min. We find three general classes of decay curves, (a) simple exponential (first order); some of these showed a shoulder before onset of exponential decay; (b) bi-component or multi-component concave upward; (c) initial stimulation of rate of mRNA synthesis, followed by virtually undetectable decay.


The half-life of mRNA in Saccharomyces cerevisiae. The decay kinetics of mRNA was studied in a yeast temperature-sensitive mutant, ts136, which is defective in cytoplasmic RNA production at 37 degree C. The disappearance of the synthetic capacity of mRNA was determined by withdrawing equal volumes of ts136 cell culture and pulse-labelling with [35S]methionine at various time intervals after the shift to 37 degrees C from 23 degrees C. The synthesized proteins were separated on a two-dimensional gel electrophoretic system and then quantitatively analyzed for theri incorporated radioactivities by scintillation counting. Our results show that yeast mRNAs have divergent functional half-lives ranging from 4.5 to 41 min, with an average value of 22 min. Each mRNA exhibits a simple exponential decay with its own characteristic dacay pattern. Of the approximately 500 major polypeptides made by yeast cells, which are detectable on autoradiograms of the gels, 80 were arbitrarily selected and the mRNAs coding for those polypeptides were examined for their decay kinetics.


A streptomycin-resistant Escherichia coli mutant with ribosomes temperature-sensitive in the suppression of a nonsense codon. Cell free extracts from a streptomycin-resistant E. coli mutant which is also temperature-sensitive for Q beta phage were studied for suppression of a nonsense mutation at various temperatures. The streptomycin-resistant ribosomes of the mutant were found to be temperature-sensitive in suppression of an amber mutation in f2 phage coat protein while retaining the ability to synthesize proteins at an elevated temperature (42 degrees C). The restriction of amber suppression at 42 degrees C is assumed to be related to an alteration in ribosomal protein S12 of the streptomycin-resistant mutant which also causes a change in its electrophoretic mobility.


The requirement of nonsense suppression for the development of several phages. A spontaneous streptomycin-resistant Escherichia coli mutant which is temperature-sensitive for suppression of a nonsense codon was studied for its ability to propagate phages T2, T4D, T5, phi K, f2, MS2, R17, Q beta, lambda as well as filamentous phages fl, fd and M13. Of all phages tested, only the growth of Q beta, lambda, and filamentous phages is inhibited in the mutant at 42 degree C. This selective inhibition suggests that, like Q beta, lambda and filamentous phages also require a read-through proten(s) which results from suppression of a termination codon.


Methylgroups of ribosomal protein L11 are not related to the synthesis of ppGpp. Ribosomes carrying either a normally methylated or an undermethylated L11, respectively, were tested with respect to the stringency reaction in the presence of crude stringent factor. Systems with either kind of ribosomes synthesize ppGpp with the same efficiency. The ppGpp synthesis in presence of ribosomes with undermethylated L11 depends also on stringent factor, mRNA and deacylated tRNA whereas aminoacyl-tRNA and peptidyl-analogous tRNA show no effect. Thus, the absence of methylation in L11 does not influence the stringency reaction.


Induced segregation in interspecific hybrids of Aspergillus nidulans and Aspergillus rugulosus obtained by protoplast fusion. Interspecific hybrids produced by polyethylene glycol induced fusion of protoplasts from auxotrophic mutants of Aspergillus nidulans and Aspergillus rugulosus were grown in the presence of the recombinogens benomyl and chloral hydrate to stimulate segregation. The A. nidulans parental strains used had a known genetic marker in each linkage group. Hybrids grown on complete medium containing benomyl yielded more segregants. Analysis of the segregants showed that the distribution of A. nidulans linkage groups was random. No specific linkage group appeared in all the segregants. The two parents are closely related taxonomically and the findings from these experiments suggest that a high degree of chromosomal homology may exist between them.


Peptide chain elongation rate and ribosomal activity in Saccharomyces cerevisiae as a function of the growth rate. The peptide-chain elongation rate of Saccharomyces cerevisiae at two different growth rates was estimated by the kinetics of radioactive labelling of nascent and finished polypeptides as described by Gausing, 1972, and Young and Bremer, 1976. The elongation rates of a diploid strain cultured in yeast nitrogen base supplemented with glucose or acetate were 9.3 amino acids/s and 5.5 amino acids/s at 30 degrees C, respectively. These data together with published values on the "ribosomal efficency" as a function of growth rate (Waldron and Lacroute, (1975) enable us to estimate the rate of synthesis of ribosomal proteins as a function of the rate of total protein synthesis, alpha r, and the fraction of ribosomes that one active in protein synthesis. We conclude that in S. cerevisiae alpha r is largely independent of the growth rate while the fraction of active ribosomes decreases with decreasing growth rate.


Cloning of an EcoRI fragment carrying E. coli tufA gene. EcoRI fragments of the transducing phage lambda fus3 DNA have been linked to the ColEl derivative plasmid RSF2124 (ColEl-Apr) DNA using bacteriophage T4 ligase. Among the plasmids formed, one designated pTUAl was found to contain the E. coli tufA gene. The proof for the presence of tufA gene in pTUAl is based on the following observations: (1) ability of pTUAl DNA and is EcoRI fragments to direct synthesis of EF-Tu in a cell-free protein synthesizing system; and (2) RNA . DNA hybridization of RNA transcribed from phage lambda rifd18 carrying tufB with DNA from pTUAl.


Sectioning of Epon blocks in the 20 m to 60 micromicron range for histological studies. Thick sections of tissue, (20 micron--60 micron), are useful in studying the relationship between individual large cells and cell layers in organized neural structures. The ability of the Nomarski Differential Interference-Contrast Microscope to bring a single thin layer into sharp focus makes the examination of such sections feasible. Although celloidin is the classical embedding medium for large, thick sections of neural tissue, the time necessary for this preparation is most inconvenient. Epon is an excellent embedding medium; however, it is extremely hard and brittle. By heating the Epon block face, thick sections can be cut. To avoid the cumbersome, often detrimental use of heat, a modification of this technique was found. Epon blocks, trimmed to a 1 millimeter square face, may be sectioned at room temperature on the sliding microtome at 20 micron to 60 micron with ease. The simple method of preparing such sections is described.


The art and science of clinical teaching. A review of the literature on clinical teaching indicates that medical schools emphasize didactic teaching of facts. Interpersonal skills, problem solving skills and cultivation of attitudes are neglected. However, the literature also described the skills of 'good' clinical teachers, and enumerates them. Programmes attempting to develop students' interpersonal skills and problem solving skills are reported. Finally, areas for further practice and study in clinical teaching are recommended.


Tinidazole in the prevention of wound infection after elective colorectal surgery. During the first six months of 1978, 71 patients were the subject of a controlled trial of the use of tinidazole for the prevention of wound infection after elective colonic surgery. The trial design was prospective, randomized and double-blind with tinidazole or placebo given at the last oral intake before operation. The objective endpoint of the trial was the presence or absence of wound infection manifested by pus. All patients underwent a standard preoperative preparation of bowel washouts, and a standardized surgical technique included, in all cases, the use of wound drainage. At the end of the trial there were three wound infections in 40 patients who were given tinidazole, and eleven wound infections in 31 patients who were given placebo. The difference in wound infection rate between these two groups is significant (x2 with Yates correction = 7.3; P less than 0.01).


Mineral springs and spring fever. Lithium: fact and fantasy in psychopharmacology. Lithium salts have been employed as medical treatment for more than two thousand years. The history of lithium in medicine reveals a strictly empiricist approach to its use which has resulted in its varying popularity as a therapeutic treatment over the past century. Much of the enthusiasm about the therapeutic use of lithium has been generated by faulty scientific process. This paper reviews the scientific errors which have surrounded the medical use of lithium in history, discusses the metaphorical use of medical terminology and the problem which is posed by the literal interpretation of such terms, and challenges the currently accepted point of view that lithium is a specific treatment for mania. The scientific errors which have resulted in the introduction of lithium as a treatment for mania are reviewed, and it is proposed that lithium treatment be viewed as the paradigm of modern psychopharmacology. The author suggests that the empiricist approach in biological psychiatry requires critical scrutiny in order to avoid tragic consequences, regarding the hazards both to patients arbittrarily exposed to lithium therapy, as well as to the scientific concept of disease as it is modified by those who wish to re-define disease, empirically, in terms of response to treatment.


Immunonephelometric assays.

Immunonephelometric assays. Nephelometry, the measurement of scattered light, determines the size, shape, and concentration of the scattering particles (at least in theory). For the applications of nephelometry in immunoassays, the scatterers are the antigen-antibody complexes formed. The rationale for using nephelometry includes the advantages of greater simplicity and improved precision as compared to other assay methods. There are, however, other factors which limit the immunological assays in which nephelometry is employed, most notably its sensitivity to noise due to dust and occasional large aggregates of antigen-antibody complexes. Nephelometers commercially available for immunoassays use a variety of light sources; most measure the scattered light at a single angle, and several incorporate microcomputers. A review of the theory of light scattering by small spheres is introduced to indicate fashions in which future developments may make nephelometry the method of choice for an even wider range of immunoassays.


Mason-Pfizer like virus in kidney grafted patients. Lymphocytes from a kidney grafted patient were specifically stimulated to incorporated thymidine in vitro by mitomycin C treated Hela cells infected with a Mason-Pfizer like virus (MPV). The thermolabile mutant of vesicular stomatitis virus (VSV tl) grown in leukocytes of this patient produced pseudotypes which were neutralized by the patient's sera and boy rabbit anti-MPV sera. Coculture of the patients leukocytes with SIRC cells yielded virus populations with dual properties; those of MPV plus those of another virus which was not typed serologically but possessed the biological properties of typical C particles. The patient also had neutralizing antibodies to MPV, which were detected by inhibition of syncytium formation on KC cells and by neutralization of VSV tl (MPV) pseudotypes. Preliminary results on the frequency of similar findings in other kidney grafted patients are presented.


The relation of repair phenomena to mutation induction in bacteria. The relation of various processes to mutation induction by radiation and chemicals is discussed for for various species of bacteria. A variety of repair processes have been identified at the molecular level that can eliminate many kinds of potentially mutagenic lesions before they can be converted to final mutation. Fixation often but not always occurs at replication. A number of mutagens, including UV light, ionizing radiation, and a number of chemicals, induce an error-prone process, perhaps a modification of the proof-reading system, that allows bacteria to survive after potentially lethal damage at the expense of making errors. Some mutagens, notably monofunctional alkylating agents and base analogues, produce mutations by other processes. Even in these cases, repair processes play an important role. There is some evidence that error-free as well as error-prone repair processes can be induced. A brief discussion is given of the relation of these findings to the practical problems of hazards estimations.


Photodynamic effects of dyes on bacteria. II. Genetic effects of broad-spectrum visible light in the presence of acridine dyes and methylene blue in chemostat cultures of Escherichia coli. Photodynamic mutagenesis was studied in chemostat cultures of Escherichia coli B/r (TlR trp) exposed to one of six different acridine dyes or methylene blue. Mutation to phage T5 resistance was induced with a broad-spectrum fluorescent-light source. All of the agents tested were photomutagenic; acridine yellow was the most efficient sensitizer and quinacrine was the least efficient. Quinacrine also was moderately mutagenic in the dark, in contrast to the other agents tested, which were not significantly mutagenic in the dark at the low concentrations tested for photomutagenesis. The mutation rate with acridine orange was directly proportional to both fluence rate and dye concentration over the ranges tested. Photomutation rates with acridine orange, proflavine and methylene blue were independent of growth rate of the chemostat cultures. These results are consistent with photomutagenesis occurring as the result of photochemical damage to DNA-dye complexes, independent of cell expression was approximately 2.5 generations for each of the photomutagens tested. This short expression delay supports an earlier segregational model for expression of phage resistance. The following results suggest that photodynamic mutagenesis is due mainly to intercalated dye molecules: (1) both acridine and 9-aminoacridine are photodynamic mutagens; (2) acridine inhibits photomutagenesis with acridine orange; and (3) neither putrescine or spermine, which bind to DNA without intercalating, inhibited photomutagenesis by acridine orange or proflavine.


[Prosthetic rehabilitation by removable prostheses with distribution of force by means of wires in Kennedy 4th class cases]. Prosthetic rehabilitation in cases of Kennedy class 4th, particularly when the from gap is extensive, presents problems of both aesthetic (lack of front teeth) and functional nature that are not easy to solve. On one side a mixed dental and mucosa support must be obtained while on the other over-loading of the abutment teeth must be avoided by distributing force axially and unformly. This is done by using a special type of prosthesis described here where force is distributed by means of wires. Cases treated like this and followed up have shown positive results as regards both clinical and mechanical results.


Effect of mutagens, chemotherapeutic agents and defects in DNA repair genes on recombination in F' partial diploid Escherichia coli. The ability of mutagenic agents, nonmutagenic substances and defects in DNA repair to alter the genotype of F' partial diploid (F30) Escherichia coli was determined. The frequency of auxotrophic mutants and histidine requiring (His-) haploid colonies was increased by mutagen treatment but Hfr colonies were not detected in F30 E. coli even with specific selection techniques. Genotype changes due to nonreciprocal recombination were determined by measuring the frequency of His- homogenotes, eg. F' hisC780, hisI+/hisC780, hisI+, arising from a His+ heterogenote, F' hisC780 hisI+/hisC+, his1903. At least 75% of the recombinants were homozygous for histidine alleles which were present on the F' plasmid (exogenote) of the parental hetergenote rather than for histidine alleles on the chromosome. Mutagens, chemotherapeutic agents which histidine alleles on the chromosome. Mutagens, chemotherapeutic agents which block DNA synthesis and a defective DNA polymerase I gene, polA1, were found to increase the frequency of nonreciprocal recombination. A defect in the ability to excise thymine dimers, uvrC34, did not increase spontaneous nonreciprocal recombination. However, UV irradiation but not methyl methanesulfonate (MMS) induced greater recombination in this excision-repair defective mutant than in DNA-repair-proficient strains. Mutagenic agents, with the exception of ethyl methanesulfonate (EMS), induced greater increases in recombination than the chemotherapeutic agents or the polA1 mutation. EMS, which causes relatively little degradation of DNA, was more mutagenic but less recombinogenic than MMS, a homologous compound ths that inhibition of DNA occurring single-stranded regions in replicative intermediates of the DNA. Mutagens which cause the rapid breakdown of DNA may, in addition, introduce lesions into the genome that increase the number of single-stranded regions thus inducing even higher frequencies of recombination.


Validation and characterization of the L5178Y/TK+/- mouse lymphoma mutagen assay system. The current status of the L5178Y/TK+/- leads to TK-/- mouse-lymphoma mutagenicity assay is described. Dose-survival-mutagenic response data are shown for 43 chemicals. Mutagenicity and cytotoxicity in the presence or absence of non-induced and/or Aroclor-induced rat-liver S-9 are compared for most of these chemicals, 25 of these for which usuable carcinogenicity data exist have been used to construct an approximately linear relationship between oncogenic potency in vivo and mutagenic potency in this system in vitro; linearity between these two endpoints extends over a greater than 100,000-fold range in potencies. Several carcinogens which are negative or difficult to detect in the standard Ames assay are mutagenic in this mammalian cell system. These include natulan, sodium saccharin (lot S-1022), p,p'-DDE (metabolite of DDT), dimethylnitrosamine, diethylnitrosamine and diethylstilbestrol. Characterization of the TK-/- mutants suggests that two mutagenic mechanisms contribute to their final yield. Large-colony TK-/- mutants probably represent point or gene mutations affecting the TK locus. In addition, a class of small-colony TK(/- mutants are described and characterized as being heritably growth-deficient; this and other properties suggest that these small-colony TK-/- mutants originate by a heritable and viable chromosomal aberration. Most carcinogens and mutagens tested produce both classes of TK-/- mutants in this system; the relative proportions of small- and large-colony mutants are both mutagen- and dose-dependent. Comparative studies have been done at the rapidly-expressing TK locus and the slowly-expressing HGPRT locus in these cells. Several carcinogens detected at the TK locus are non- or very weakly mutagenic at the HGPRT locus. This findings is consistent with the induction of slow-growing specific locus mutants by a chromosomal mechanism and their subsequent dilution during this long expression time.


Photodynamic effects of dyes on bacteria. III. Mutagenesis by acridine orange and 500-nm monochromatic light in strains of Escherichia coli that differ in repair capability. In the presence of acridine orange (AO) and monochromatic 500-nm light, the recombination-deficient strain of Escherichia coli, WP10 (recA), showed a 15-fold increase in mutation rate over the wild-type (WP2) strain. Under the same conditions, strain Bs--1 (uvrB lexA lon) showed a 5-fold increase in mutation rate over strain WP2. In contrast, the endonuclease-deficient, strain, WP2s (uvrA), showed a lower AO-500 nm mutation rate than wild-type. The extremely high mutation rate of the recA strain cannot be due to error-prone inducible SOS repair since the inducible recA + function is absent. Repair of the AO-500 nm-induced lesions is likely due to a recA+-dependent, error-free, recombination process. It is concluded that the high mutation rates with AO-500 nm light obtained in chemostat cultures of recA and lexA strains occur as a consequence of errors during semi-conservative DNA replication in the presence of unrepaired DNA lesions.


Mutation induction by thymidine deprivation in Escherichia coli B/r. I. Influence of caffeine. The addition of caffeine to the plating medium after thymine deprivation of E. coli WP2 uvr+ thyA or WP2 uvrA thyA had no influence on survival. Caffeine, however, reduced the frequency of mutants. The hypothesis is presented that the reduced mutagenesis is due to the sensitivity to caffeine of an inducible error-prone repair mechanism operating during thymine deprivation and after the re-addition of thymine.


Aryl-monoalkyl and cyclic triazenes: direct-acting mutagens. An aryl-monoalkyl triazene, methyl-p-tolyl triazene (MTT) and a cyclic triazene (delta2-triazoline) are direct-acting mutagens for Salmonella typhimurium bacteria and for cell-free Hemophilus influenzae DNA. MTT causes reversion of the hisG46 base-substitution mutation, but no reversion of the hisD3052 frameshift mutation. Induced mutation frequency is not strongly influenced by modifications in the genetic background of the S. typhimurium Ames tester strains, but is mildly enhanced by the addition of a pool of amino acids to the plating medium and is strongly enhanced by liquid preincubation before plating.


Effect of gamma and alpha irradiation on survival of wild-type and sensitive mutants of yeast. Survival curves have been obtained for haploid and diploid Saccharomyces cerevisiae of the wild-type strain and the sensitive mutant rad51-1 exposed to gamma-rays and alpha-particles. The difference in radio-sensitivities of the wild-type diploid strain and the homozygous rad51-1 mutant was significantly higher after low-LET radiation than after high-LET radiation. This was not true for haploid strains. The liquid-holding recovery of irradiated cells was greater for gamma-ray-induced damage than for damage inflicted by alpha-particles. The value of the RBE of high-LET radiation was correlated with cell-repair capacity.


Further study of the genetic toxicity of gentian violet. The genetic toxicity of gentian violet was studied with the Ames and the Rosenkranz bacterial assays as well as the cytogenetic assays (Chinese hamster ovary cells in vitro in the presence of rat-liver S-9 fractions, the chicken-embryo and mouse-bone-marrow cells in vivo). Gentian violet was found to be toxic but not mutagenic in the Ames assay. However, it was active in the Rosenkranz assay causing reparable DNA damage. The presence of S-9 in the in vitro cytogenetic assay and in the bacterial assays showed that the activity of gentian violet could be reduced or eliminated. In the in vivo assays, gentian violet was not clastogenic and failed to induce sister-chromatid exchanges. However, gentian violet proved to be highly toxic to growing chick embryos at high dosage and depressed mitotic activities in mouse bone marrow after prolonged treatment. Our study suggested that gentian violet can be inactivated by the liver detoxification system. However, it is potentially hazardous to cells that are exposed to the dye directly (e.g. skin epithelium and cell lining of the gastrointestinal tract).


Actions of an antispermatogenic, but non-mutagenic, indenopyridine derivative in mice and Salmonella typhimurium. This paper describes a new antispermatogenic agent. Following single oral administration to mice, the indenopyridine derivative (4aRS,5SR,9bRS)-2-ethyl-1,3,4,4a,5,9b-hexahydro-7-methyl-5-p-tolyl-2H-indeno(1,2-c)pyridine hydrochloride, code No. 20-438, produced long-lasting inhibition of the spermatogenic process at dose levels of 10 mg/kg (1/40 of the lowest lethal dose) and higher. Testes weights were significantly reduced from days 2--217 after treatment, and no clear-cut evidence of a recovery was found during this time. The fertility of treated males was normal during the initial 2 weeks after treatment, followed by partial or total sterility in weeks 3--6, and incomplete recovery in weeks 7--29 after treatment. The antifertility effects were caused by maturation depletion of the germ cells, leading to oligospermia. The following rank of decreasing "susceptibility" of the germ cells was observed: Spermatocytes greater than early spermatids, intermediate spermatogonia greater than stem cells. Sperm and late spermatids were not affected. Despite the characteristic specific germ-cell pattern of antifertility effects, 20-438 showed neither indications of pre- and post-implantational dominant lethality, nor mutagenic potentiality as measured by cytogenetic analysis of spermatocytes or spermatogonia, the sperm abnormality assay, the micronucleus test, and the Salmonella assay. These data suggest that the action of 20-438, leading to oligospermia, does not involve genetic toxic effects.


Lack of cytogenetic effects in mice or mutations in Salmonella receiving sodium fluoride. We have examined the possible effect of fluoride intake on chromosome damage. There was no evidence of increased frequency of chromosomal aberration in bone marrow or testis cells of mice with either 50 ppm fluoride intake over several generations or 100 ppm intake for 6 weeks compared to animals drinking distilled water. Fluoride was not found to be mutagenic in a widely used bacterial mutagenesis assay over a range of 0.1 to as high as 2000 microgram fluoride per plate.


Skeletal concentrations of lead in ancient Peruvians. The level of biologic lead (expressed as the ratio of atomic lead to atomic calcium) in bones of Peruvians buried 1600 years ago was found to be 3 x 10(-8), as compared to 2100 to 3500 x 10(-8) in the bones of present-day residents of England and the United States. The ratio of barium to calcium was 2 to 3 x 10(-6) in bones of ancient Peruvians and present-day Americans. Barium and lead have similar morphologic distributions in organisms, so this discrepancy for lead must result from overexposure of present-day people to industrial lead and not from natural variations. The magnitude of this discrepancy has been confirmed by two different lines of investigation not reported in this article. This new evidence suggests that natural interactions of lead in human cells have not yet been determined because reagents, nutrients and controls used in laboratory and field studies have been contaminated with lead far in excess of naturally occurring levels.


The ribosomal RNA gene region in the sea urchin Lytechinus variegatus. A fine structure map of the ribosomal gene region of the sea urchin Lytechinus variegatus has been generated. The ribosomal DNA density satellite has been analyzed with restriction enzymes and by hybridization to urchin 18S and 26S ribosomal RNA (rRNA). The L. variegatus rRNA repeats show heterogeneity on the nontranscribed spacer. Minor bands containing transcribed spacer sequences are observed. One interpretation of these bands would require both transcribed spacer heterogeneity and defective ribosomal genes retained among the rRNA gene repeats.


Genetics of cultured mammalian cells, as studied by somatic cell hybridization. In the past 10 years, the technique of somatic cell hybridization has been used extensively in studies on mammalian somatic cell genetics. Much of the work with hybrid cells has focused on the problems of gene regulation and gene mapping; these two areas and the consideration of new techniques are discussed in this article. The types of genetic manipulations than can be used to investigate these problems are the central theme of the review.


In vitro carcinogenesis with cells in early passage. A well-documented rationale exists for the study of the induction of cancer at the cellular level. Transformation can be quantitated; its frequency follows a linear relationship with dose and is consistent with a "one-hit" phenomenon. Transformed colonies do produce transformed lines with attributes of neoplastic cells including the production of tumors; in vitro activity correlates with in vivo activity to provide evidence that chemically induced carcinogenesis can be studied in vitro. In vitro techniques utilizing mammalian cells in culture have made possible the rapid evaluation of carcinogenicity of agents in man's environment. Neoplastic transformation is inductive and not the result of the selection of preexisting tumor cells. The addition of a host-mediated step in the bioassay makes it possible to decrease the number of false negatives, which may result from the requirement for metabolic activation of the chemical. Thus the in vitro studies described have a high probability of providing practical methods for determining which chemicals in use have a potential of producing cancer. Furthermore, the nature of the cell-target insult interaction can be determined, as well as the chemical nature of the ultimate carcinogen, the degree to which any agent acts alone, be it a chemical, a virus, or irradiation, and the extent to which one agent interacts with another from the same or a different category of carcinogens. Sequential treatment involving chemicals, viruses, and radiation are important, since combinations of various agents may be responsible for an increased risk of cancer in laboratory animals and human populations. The use of multiple agents may also lead to different but specific new types of assays to use for surveillance of our environment for carcinogenic agents. Pretreatment of Syrian golden hamster embryo cells with either X-irradiation or methyl methanesulfonate, but not UV-irradiation, increases the frequency of chemical transformation as does posttreatment with caffeine. Most, if not all, chemical carcinogens will increase the sensitivity of hamster embryo cells to transformation by a carcinogenic simian adenovirus SA7. The enhancement of virus transformation is related to both the length of chemical treatment and the interval between chemical and viral addition. The mechanism of transformation enhancement by various agents has yet to be explained. They may affect a number of molecular processes or cause a modification of existing DNA and thus provide an explanation for carcinogenesis; in fact, in some systems some of these agents may also show mutagenic activity and produce chromosomal aberrations, However, although DNA is the critical site for a mutagen, the critical target(s) of chemical carcinogens is still unknown.


Metabolism of acetylcholine receptors in skeletal muscle. The acetylcholine receptor in skeletal muscle is an integral plasma membrane glycoprotein. Its biosynthesis and incorporation into plasma membrane and its degradation are being studied with the use of biochemical, biophysical, and microscopic techniques. In this report, previously published data are combined with new information to yield a consistent and fairly detailed description ofthe mechanisms involved in receptor metabolism. It is proposed that the biosynthesis, transport, and incorporation of the receptor into plasma membranes involve a mechanism similar, or identical, to that used by the cell for production and secretion of secretory proteins. The receptor is degraded by a random-hit process, which involves internalization, transport to secondary lysosomes, and hydrolysis. Sites of regulation of receptor metabolism are discussed in the context of regulation of the number and distribution of receptors in plasma membranes, particularly with respect to the formation and stability of neuromuscular junctions.


The genetics of the mitochondrial DNA of mammalian somatic cells, their hybrids and cybrids. A brief review is attempted of the status of the genetics of the mitochondria of mammalian cells in vitro. Reverse segregant human-mouse hybrid strains are unusually stable and retain mitochondrial DNA (mtDNA) of both species for many generations but eventually lose the mtDNA of the species whose chromosomes are segregated. A molecular interchange resembling recombination among mtDNA molecules can be detected in these hybrids. Eisenstadt and co-workers have shown that chloramphenicol inhibits protein synthesis less in mitochondria isolated form chloramphenicol-resistant, mutant cell strains than from sensitive strains. They have also shown that resistance is effectively transmitted by cytoplast fusion. These genetic markers are, therefore, probably in the mtDNA of mammals as they are in yeast. Although chloramphenicol resistance has not been transferred from one distinct species to another by cytoplast fusion, we find that different subspecies of mouse are able to exchange resistance. We have also transferred chloramphenicol resistance to ("transformed") 10(-3) sensitive cells by treating them with purified mtDNA from resistant cells. Treatment with DNA from Escherichia coli and/or mtDNA purified from sensitive cells does not transform. Treatment with nuclear DNA, however, from either sensitive or resistant cells does transform. We suggest that exogenously applied nuclear DNA acts as a potent mutagen in mitochondria. A potential application of the mtDNA transformation phenomenon to molecular cloning of DNA in mammalian mitochondria is proposed.


Red cell-mediated microinjection.

Red cell-mediated microinjection. I have reviewed the current status of microinjection based on fusion of red blood cells and tissue culture cells. Macromolecules are introduced into red blood cells during hypotonic hemolysis, and the resealed red cells are then fused to tissue culture cells with Sendai virus. The procedure has been used to inject ferritin, thymidine kinase, bovine serum albumin, and transfer RNA molecules into large numbers of tissue culture cells. Physiologically significant amounts of various macromolecules can be transferred, and preliminary studies show that [125I]bovine serum albumin and transfer RNA are stable within recipient culture cells. Tissue culture cells remain viable following microinjection. Red cell-mediated microinjection should facilitate the study of various processes, such as macromolecular turnover and genetic regulation, that are not easily studied with conventional biochemical techniques.


Transitional cell cancer: fluorescent antibody binding to tumor cells in vitro. The IFA assay has been used to detect antigen--antibody reactions between sera from patients with TCC and TCC cell lines established in our laboratory. A positive immunofluorescence reaction, consisting of perinuclear staining, was seen in 86% of the sera from TCC patients exposed to 253J cells. A positive reaction was detection in only 8% of the sera from healthy donors, in 19% of that from patients with nonmalignant urinary tract disease, and in 31% of the sera from patients with urinary tract tumors other than TCC when exposed to 253J cells. No positive reaction was detected when these same sera were exposed to either WI-38 or normal human kidney cells. The positive fluorescence could be removed from TCC sera by absorption with TCC surgical specimens but not by absorption with nontumor surgical specimens.


Identification of cultured, human, malignant, prostatic epithelial cells. Identification of cloned cells is necessary for experimentation with them. This paper details a method for the identification of cultured human malignant prostatic epithelial cells derived from metastatic deposits of prostate cancer by localization of a specific rabbit antiserum to human prostatic acid phosphatase in the cells.


Poly I:C immunotherapy in patients with papillomas or superficial carcinomas of the bladder. A randomized prospective therapeutic trial of poly I:C immunotherapy in patients with papillomas or superficial carcinomas of the bladder is presented. Poly I:C had no beneficial effect on the recurrence rate of superficial bladder tumors.


EB 33, an epithelial cell line from human prostate carcinoma: a review. The cell line EB 33 was cultivated from the tissue of a human prostate adenocarcinoma in June 1973. Since that time, efforts have been made by scientists in several laboratories to characterize this line by morphologic, biochemical, endocrinologic, genetic, and immunologic parameters. The great need for a well-defined prostatic cell line for immunologic experimentation warrants a complete review of the subject.


Transmission of allosteric effects in DNA. Binding of distamycin induces a cooperative transition of calf thymus DNA to a new form with higher affinity for the drug and altered structural properties.


Distribution of polymorphic forms of troponin components and tropomyosin in skeletal muscle. Specific antibodies have been used to show that in adult skeletal muscle the slow and fast forms of the components of the troponin complex are located in type I and type II fibres respectively. alpha-Tropomyosin is restricted to type II cells. During development, as a result of changes in innervation and in certain diseased stages, both the slow and fast polymorphic forms of the troponin components are present in the same cell.


Experimental data concerning the question of the synergic action of the influenza virus and of some chemical carcinogens in the pathogenesis of lung cancer. The importance of new discoveries in the field of molecular biology concerning the interaction of oncogenic material in cell leading to a synthesis of knowledge in the field of viral and physico-chemical blastomogenesis is underlined. A concise reference to earlier studies and to the present state of knowledge is presented, with a subsequent review of the results of experimental studies dealing with the problem of the synergic action of influenza viruses (type A2, A/PRS, B, Sendai) and of chemical carcinogens (urethane, cancerogenic hydrocarbons, diethylnitrosamines, cigarette smoke) in the pathogenesis of lung cancer. Both the suggested and already demonstrated mechanisms of the joint action of viruses and chemical carcinogens, illustrating the interrelations between the two factors in the process of malignant transformation, are discussed.


Splenic and thymic histamine forming enzyme activity and weights following organ allograft and tumor cell transplantation. Kinetic of the increasing activity of histamine forming enzyme--histidine decarboxylase in host spleen and thymus, following kidney and heart allografting and after tumor cells transplantation is presented.


Antitumor effect of sublethal irradiation caused by its tolerance abrogating capacity. After the resection of methylcholanthrene-induced fibrosarcomas, representing 1--2% of total body weight, "R" rats were immunized with glutaraldehyde-fixed tumor cells, irradiated sublethally [0.1419 C kg-1 (550 R)], and restored immunologically by thymus, spleen and bone marrow cells. Afterwards, 87.5% of them were able to reject a viable challenge cell graft of 1 X 10(5) cells. Sublethal irradiation alone had the same effect, reflected by 90% of rats rejecting the grafts. Five control lots were run. They showed that none of the treatments, applied to "tumor-resected" animals could provide normal animals with the same defense capacity. Results point to the decisive role of the transient presence of the tumor in the host and of the sublethal irradiation in restoration of its defense capacity. Role of partial tolerance, in producing the host's immune inhibition, and of the capacity of irradiation to abrogating it, are discussed.


Luteinizing hormone-releasing hormone (LH-RH) neurons in cultures of fetal rat hypothalamus. Hypothalamic fragments from 21-day-old fetal rats were cultured in Maximow double-coverslip assemblies for 1 to 2 months. Neurons containing LH-RH were demonstrated immunohistochemically using an antiserum to LH-RH (Dermody; 1:500--1:4,000). LH-RH was demonstrable only in neuronal perikarya (8--13 micrometer) and in small (less than 1 micrometer) round structures nearby, primarily in explants of the median eminence-arcuate nucleus region. Reactive neurons were not found in explants of the preoptic area and could not be demonstrated in fetal hypothalami at the time of explantation. The presence of mature-looking LH-RH containing neurons in these cultures suggests that this tissue culture system can be used for the study of hypothalamic development.


The profile and severity of lithium-induced side effects in mentally healthy subjects. In a double-blind, placebo-controlled, crossover design various side effects (= unwanted effects) during 6 months of therapeutic serum lithium levels (0.7-1.1 mmol/1) to nonpsychiatric patients were studied. The side effects were measured by self-rating scales and independent observer rating scales administered every 2-4 weeks throughout the study. After months of treatment, lithium induced hand tremor and thirst/polyuria; however, without any relationship to the serum levels of lithium. The frequency of tremor was highest in patients above the age of 60 years. No initial sedative-like lithium effect was found.


Creutzfeldt-Jakob disease: familial clustering among Libyan-born Israelis. Creutzfeldt-Jakob disease was studied among Libyan-born Israelis, in whom the disease appears with unusual frequency. Interviews with relatives of deceased victims revealed statistically significant clustering within families. The results suggest either a common source of exposure or a genetic influence on susceptibility to the virus.


Paying for physician services under Medicare and Medicaid. Public systems for physician reimbursement aim to reconcile two disparate objectives: ensuring availability of services to the poor and aged; and keeping rates of cost increase within acceptable limits. Several interesting--and unorthodox--policy simulations of physician pricing behavior are investigated through econometric estimation. Current arrangement for paying physicians are fraught with difficulties. The objectives of Medicare and Medicaid are not well served.


[The effect of respiratory mechanics on the hematocrit. III. Study in patients with bronchopulmonary disease treated with intermittent positive pressure inhalation therapy]. The Authors studied the behaviour of lung volumes and mechanics and of haematocrit after intermittent pulmonary positive pressure breathing (IPPB) in patients with chronic obstructive lung disease. An improvement of obstructive sindrome at 10'-20'-40' and a contemporary decreasing of haematocrit were observed. This statistically significant modification was regarded as an expression of a different distribution of blood in lung vessels dued to an increasing of this vascular compliance and to a decrease of alveolar inflation. The Authors postulate the realization of a better circulation in lung vessels which contributes to a lowering of haematocrit in these patients perhaps for a different VA/Ob ratio.


[Hepatic porphyria].

[Hepatic porphyria]. Porphyria is making increasing demands on the attention of clinicians and research worker. An account is given of hepatic forms, since these have recently come into prominence on account of recent advances in the understanding of their metabolic, diagnostic and therapeutic aspects. A description of the physiopathology of porphyrin metabolism is followed by an examination of the incidence, genetic features, aetiology, pathogenesis, pathological anatomy, symptomatology, diagnosis, prognosis, and treatment of each form. Particular attention is devoted to intermittent acute and cutanea tarda porphyria, since these are more commonly encountered in practice. Personal experience gathered in a large series of cases of cutanea tarda porphyria is presented.


[Antitubercular chemoprophylaxis after 20 years of its world-wide application]. The researches carried out on calves (1952--1953) and over 3000 guinea-pigs, rabbits and rats in the Forlanini Institute of Rome for five years, are remembered. The first clinical experiment on a group of 500 children exposed to family contagion (1955--1956) is also related. The links between INH-prophylaxis and anti-tuberculous immunity, and between INH-prophylaxis and vaccination with BCG are discussed. The clinical indications of this method among miners with pneumoconiosis, in the psychiatric institutions, and among patients subjected to long cortisone treatment are exposed. A short synthesis of the favorable results obtained in many countries everywhere is reported. Lastly the Author synthesizes the vast experiment carried out during this decade by I.U.A.T. on the remarkable efficacy of INH-prophylaxis among adults with fibrotic lung lesions in seven nations in Europe. The Author concludes the paper, affirming that chemoprophylaxis with isoniazid, after 20 years of application, has obtained the official chrism and can furnish, besides the BCG vaccine, a remarkable contribution to the control of tuberculosis in all countries of the world.


[Diagnosis of chronic reflux esophagitis. Role of endoscopic and histological examination]. Fibroendoscopic and histobioptic study of the distal oesophageal mucosa has been carried out in a series of patients suffering from gastro-oesophageal regurgitation. The regurgitation condition was verified and its extent established by means of anamnestic, spot-fluorographic and manometric-pHmetric investigations. On the basis of the analysis of relations between the extent of the oesophagitis assessed on the basis of endoscopy and that arising out of microscopic examination of the biopsy, it is concluded that there is no complete correspondence between the two techniques insofar as endoscopy can give false positive or false negative results. On the basis of these results and of those of functional investigations, histology is considered desirable even when the oesophageal mucosa is normal, if there are clinical and/or laboratory signs of gastro-oesophageal regurgitation.


[Functional study of the pancreatic alpha and beta cells of normal aged subjects, using a standard meal]. Modifications in glycaemic, insulinaemic and pancreatic glucagon levels have been studied in 20 normal elderly subjects and in 20 young adults (controls) following a standard meal. Results showed that glycaemia is higher and insulinaemia rises more slowly in the elderly, while serum glucagon values show no substantial variations in the two groups of subjects.


[Anti-ammonemic activity of the gamma-ethyl ester of glutamic acid. Controlled clinical study]. A controlled clinical trial was made of the anti-ammoniaemic properties of gamma-ethylester glutamic acid, arginine and placebo in a series of 45 patients with hepatic insufficiency characterized by hyperammonemia (excluding cases of pre-coma and coma) randomly assigned to three groups for treatment with: gamma-ethylester of glutamic acid (25 g), l-arginine (25 g) and physiological solution in identical 500 ml bottles. Various parameters were controlled. The results obtained were assessed with different statistical techniques, depending on the parameter concerned and the aims of the trial. Blood ammoniaemia was significantly decreased by both-ethylester of glutamic acid and arginine though the former was more effective quantitatively and in terms of time. SGOT and BSF values also followed the same pattern.


Creutzfeldt-Jakob disease--alteration in ganglioside sphingosine in the brain of a patient. Gangliosides isolated from the brain of patients with Creutzfelt-Jakob (C-J) disease were analyzed. The ganglioside current was abnormally low, and the percentage distributions of individual gangliosides and the long-chain base compositions were abnormal. The C20-sphingosine contents of all the ganglioside fractions were low. Abnormalities in ganglioside long-chain bases in adult human brain have been reported previously only in patients with inherited metabolic disorders. These abnormalities in C-J disease seem to be closely related to the cause of the disease.


Bone and joint complications of maintenance haemodialysis and renal transplantation. Bone and joint complications of haemodialysis and renal transplant managed at Auckland Hospital have been reviewed. All pathological fractures in patients on haemodialysis united with conservative treatment and a cervical spine infection resolved following currettage of the lesion. Avascular necrosis was the main complication following renal transplant, the earliest pathognomonic radiological sign being a lucent line parallel to the articular surface, although this was often preceded by symptoms. Conservative management successfully controlled symptoms of femoral condylar and humeral lesions but only temporary relief was gained when the hip was involved. Total hip replacement, however, gave good results in all cases (10 hips) without significant operative complications.


Diclofenac in rheumatoid disease.

Diclofenac in rheumatoid disease. The clinical activity and side effects of a new phenyl acetic-acid derivative, diclofenac, have been assessed in 24 patients with rheumatoid disease. Diclofenac 25 mg q.i.d. was compared with 750mg of aspirin BP q.i.d. using a double-blind crossover technique involving four weeks on each drug. Benefits were similar for the two preparations but with improvement in morning stiffness and incidence of dyspepsia favouring diclofenac, and a more marked reduction of the sedimentation rate occurring with aspirin. A moderate elevation of liver enzymes was seen in one patient on diclofenac, and of lactate dehydrogenase in one other. It is concluded that diclofenac is a therapeutically active analgesic anti-inflammatory agent.


Dysuria in general practice.

Dysuria in general practice. An analysis of 98 clinical consultations for dysuria in an urban general practice revealed that only 60 percent of patients had proven urinary tract infections. Two percent had gonorrhoea and the rest had an urethral syndrome. Most of the urinary tract infections were caused by Escherichia coli and Streptococcus epidermidis. Proteus mirabilis was the next commonest infecting organism. Among the commonly used urinary antimicrobials, nitrofurantoin was found to be the cheapest and most effective agent. The aetiology of the urethral syndrome was obscure and apart from a small minority of patients who had thrush (16 percent) no obvious cause could be found.


Thrombocytopenia in obstetric syndromes. A review. Thrombocytopenia is a phenomenon sometimes observed in connection with various obstetric syndromes. Many times it appears clinically to be a comparatively benign complication of an otherwise significant disease. In some cases, however, it may be a clue to serious disease and its pathophysiology. The subject of thrombocytopenia induced by or related to various obstetric syndromes is reviewed and comments offered regarding the significance of the observation of platelet deficiency in these conditions.


Contemporary management and conduct of preterm labor and delivery: a review. Current concepts in the conduct of preterm labor and management of delivery of the premature infant are reviewed. Pharmacologic modalities available for inhibiting preterm labor are discussed as well as the efficacy, indications and contraindications for these agents. An analysis of the role of corticosteroids in achieving fetal pulmonary maturity in the preterm infant is reviewed, and based on the existent literature the intrapartum management of the small fetus is outlined. The mode of delivery, vaginal or via cesarean section, for these infants is likewise discussed.


The role of C. albicans in denture stomatitis. Denture stomatitis is an erythematous disorder of the denture-bearing tissues, especially the maxillary hard palate. Frequently it mimics many oral lesions. Early diagnosis of the lesion is essential to assure rational therapy. A diagnostic procedure based upon the enumeration of Candida albicans appeared to be feasible. A control group of twenty fully dentate subjects with no clinical evidence of stomatitis was sampled to establish normal concentrations of C. albicans in the oral flora. Samples were taken from a triangular area of the hard palate, serially diluted in broth, and all dilutions were incubated aerobically. In the control population, C. albicans never exceeded 100 organisms per square centimeter per subject sample. A population of edentulous patients with denture stomatitis, prior to any therapy, was similarly sampled. In many of these patients the concentration of C. albicans always exceeded 10,000 organisms per square centimeter per patient. The patients received denture replacements and antimycotic antibiotic therapy. Follow-up cultures were taken 4 to 6 weeks after completion of therapy. All patients were found to be free of denture stomatitis.


The effects of etching enamel with acid on human dental pulp. A preliminary study. Enamel acid-etching has become a widespread, popular clinical procedure in recent years. Numerous studies have investigated the nature of the acid-treated enamel. However, little or no information is presently available pertaining to the effects of acid treatment upon dental pulp tissue. A limited investigation was carried out on bilateral orthodontic extraction cases to determine the effects of enamel acid-etching upon healthy dental pulp. In each of five cases one premolar served as the experimental tooth, while the contralateral premolar served as the control. After a 24-hour postoperative interval there appeared to be no basic histologic difference between the experimental and control groups.


Progress in community pediatric research. This review examines three major facets of progress in community pediatric research. The number of publications with this orientation has increased substantially over a 10-year period, and abstracts submitted to the Ambulatory Pediatric Association meetings have doubled. New methodologies are identified and their application to ambulatory pediatric care is described. In particular, procedures and approaches are examined in the context of recognized criteria for sociomedical research.


The identification of soluble adult antigen on the tegumental surface of juvenile Fasciola hepatica. An antiserum was raised in rabbits against a soluble extract of fresh homogenized adult Fasciola hepatica of rat origin and was then absorbed with rat and mouse tissue antigens. This antiserum reacted specifically with the surface coat of adult flukes, of both rat and mouse origin, by indirect immunofluorescence to show the detail of surface spines. When tested against juvenile stages recovered from mice the reaction was positive with all but the earliest hepatic parenchyma stages. No reaction was present on the tegumental surface of newly excysted juveniles or stages 1 or 2 days post-infection (p.i.) whether recovered from the peritoneal cavity or the hepatic parenchyma.


The ribosomes of Plasmodium berghei: isolation and ribosomal ribonucleic acid analysis. Ribosomes and high molecular weight ribosomal ribonucleic acid (rRNA) from the blood stages of Plasmodium berghei parasites were studied in preparations free from host ribosome contamination. Purified malarial ribosomes were isolated in high yield from a population of ultrastructurally intact, viable parasites by hypertonic lysis with Triton X-100 and differential centrifugation. These ribosomes were shown to be derived from active polysomes and could be dissociated into subunits by puromycin-0.5 M KCl treatment. Malarial rRNA extracted from purified 40S and 60S ribosomal subunits was characterized by electrophoretic, sedimentation and base ratio analyses. Like certain other protozoa, the P. berghei 40S ribosomal subunit possessed an exceptionally large RNA species (mol. wt 0.9 X 10(6), while RNA isolated from the parasite's 60S subunit (mol. wt 1.5 X 10(6)) was specifically 'nicked' to produce one large component (mol.wt 1.2 X 10(6)) and one small component (mol.wt 0.3 X 10(6)) in equimolar quantities. These rRNA's migrate identically on polyacrylamide gels after heating to 63 degrees C for 5 min or under denaturing conditions in the presence of formamide, indicating an absence of aggregation and non-specific degradation of the rRNA species. Base composition studies showed P. berghei rRNA to be low in guanosine and cytosine content, as is the case for protozoa generally.


[Ultrastructural study of rat liver after administration of an aminated steroid (author's transl)]. The administration of amino-3 beta hydroxy-20 beta pregnene-5, to the male Wistar rat, per os, at doses of 100 and 200 mg/kg/24 h, induce the development of a chronic active hepatitis. The ultrastructural observation shows slight changes only in perilobular hepatocytes at the beginning of treatment; then hepatocellular alterations progressively increase and may be observed in the whole lobule after 40 and 80 days of treatment; the progression of hepatocellular damage is associated with collagen increase and bile duct proliferation. The interest of this experimental hepatitis, as a model analogous to human chronic active hepatitis, is discussed.


Biochemical studies on glomerular basement membrane in human diabetic microangiopathy. Biochemical modifications of glomerular basement membrane (GBM) in diabetes mellitus have been investigated in human subjects. Amino acid analysis and electrophoretic study of unfractioned GBM from diabetic and non diabetic kidneys showed an increase in hydroxylysine, hydroxyproline and glycine and in the glycosylgalactosyl-hydroxylysine (G-G-H) content of diabetic GBM. A decrease in 1/2 cystine was also demonstrated in diabetes. Fractionation of GBM showed the increase in G-G-H observed on unfractioned GBM to reflect a quantitative increase of the "collagen" fraction in diabetes, the amino acid composition of this fraction remaining unchanged. The decrease in 1/2 cystine demonstrated on unfractioned GBM was located in the non collagenous fractions of this structure.


[Study of amino monosaccharides by thermoionic detection in gas-liquid chromatography. Application to a pyroglobulin IgM (author's transl)]. The monosaccharides of proteic samples liberated by methanolysis and submitted to trimethylsilylation by the trimethylsilylimidazole (TSIM) are determined by gas-liquid chromatography. The neutral sugars are detected in simple flame ionization (FID) and the amino sugars identified separatively by the use of a thermoionic detector (TID) which responds selectively to nitrogenous and phosphorated compounds. The method described has been tested on standard mixtures of monosaccharides and on a well known glycoprotein : the uromucoid. Then it has been applied to a pyroglobulin IgM (mu2, chi2).


[Lipid particles agglutinated in hyperlipidemias with opalescent sera (author's transl)]. During the five last years, we observed under the microscope all the hyperlipidemic sera received in our laboratory, to search a lipid particle agglutination. We have retained 24 sera with obvious agglutination (9 types V, 9 types IV and 6 types IIb), the lipidemia of which was compared, type to type, to 56 control sera (16 types V, 16 types IV and 24 types IIb), in which no agglutination has been found. We observed significant differences in the means of cholesterol (CT) triglycerids (TG), vitamin A (VA) and CT/TG, VA/TG and VA/CT ratios, comparing agglutinated and control sera. These differences suggest that the agglutination phenomenon is in relation with the nature of the lipid particles present in the hyperlipidemic sera, which could be "chylomicron remnants" in type V hyperlipidemias, and other forms of "remnants" in type IV and IIb hyperlipidemias. These "remnants" could result from a blockade of lipolysis. This blockade of lipolysis in the agglutinated sera might be related to what is known about the authentificated autoimmune hyperlipidemia (AIH). Thus we can suppose that hyperlipidemias with obvious serum lipid particle agglutination are autoimmune hyperlipidemias.


[Investigation of circulating immune complexes in some diseases associated with a platelets hyperdestruction process (author's transl)]. A possible relationship between the presence of circulating immune complexes (CIC) and the platelet-bound IgG levels was investigated in 38 patients with some diseases associated with a no drug-related platelets hyperdestruction process of which 33 had idiopathic thrombopenic purpura (ITP). By using the 125I-C1q binding test, these CIC were only detected in 4 of the 42 tested sera. No correlation was noticed between the percentage of C1q binding and the platelet IgG levels evaluated by the quantitative hemolysis inhibition test. These results do not allow to consider the involvement of CIC in the thrombocytopenia genesis observed in these diseases.


[Pathophysiology of chronic, bronchial hypersecretion (author's transl)]. The biochemical and rheological analysis of sputum allows one to detect abnormalities of the bronchial secretion and to describe the development of the bronchial disease. The main biochemical components of sputum and their biological activities are first described. The fibrillar structure of the mucus is related to heavy weight glycoproteins and these macromolecules contribute to the viscoelastic properties of the bronchial secretion, by their capacity to combine with other proteins such as secretory immunoglobulin A (S-IgA). As the chronic bronchial hypersecretory state progresses, the glandular synthesis of mucins and/or secretory proteins decreases while the passive filtration of blood-derived proteins, such as serumalbumin, increases. Such biochemical changes are frequently associated with a rise in viscosity and a loss in elasticity of sputum. These rheological modifications result in an impairment of the mucociliary clearance and therefore contribute to airway obstruction. The assessment of different indices such as the equilibrium in mucin secretion, S-IgA/serum-albumin ratio, viscosity and elasticity of sputum and its mucociliary transport rate, should lead to a better knowledge of the defense properties of the bronchial mucosa.


[Spectrofluorimetric automatised assay of free and total tryptophan in plasma (author's transl)]. An automated fluorometric method, including norharmane transformation of tryptophan (TRP) by formaldehyde and perhydrol, is applied to free and total TRP determination in human plasma. Separation of free TRP determination in human plasma. Separation of free TRP is achieved by ultrafiltration at pH 7,25 under an air/CO2 flow. The procedure here described is easy, rapid and very sensitive, allowing to work on microsamples if free TRP separation could be realized with them. A correct precision is obtained (CV : 2,8% for plasma repetability) and the specificity is quite good in plasma according to the negligible effect of tryptamine. Frequent values, calculated from 19 healthy subjects are [mean +/- tSE (risk 5%)]:--free TRP : 11,8 +/- 1,5 mumol/l;--total TRP : 52,2 +/- 19,7 mumol/l.


Retinoblastoma.

Retinoblastoma. Retinoblastoma is a radiosensitive malignancy of neuroblastic origin that primarily affects young children. Its relatively low incidence belies its potential importance in the understanding of tumor biology in general. A case is made for referral of all retinoblastoma patients to centers with retinoblastoma protocols.


Right aortic arch with aberrant left innominate artery. A 9-year-old girl with type D right aortic arch anomaly, has been presented. The diagnosis was based upon the presence of signs and symptoms of a vascular ring and the early visualization of a retroesophageal aberrant left innominate artery, by means of countercurrent right brachial angiography. Division of the ligamentum arteriosum at operation relieved the symptoms.