[Pathology of mastitis in cows, goats and swine. A review]. The literature concerning pathogenesis, pathological physiology and pathological anatomy in bovine, caprine and ovine mastitis is reviewed. The evoluation of infectious mastitis is divided in the three following phases: transmission-, invasion- and establishphase. The pathological anatomy about Mastitis acuta haemorrhagica et necroticans, Mastitis catarrhalis acuta, Mastitis catarrhalis chronica, Mastitis suppurativa and Mastitis granulomatosa is discussed.


[Closure of the vagina after colpohysterectomy using an automatic suture forceps (author's transl)]. Closure of the lower third of the vagina using a TA 55 automatic suture forceps is an easy procedure which offers the best guarantee from a septic and haemostatic standpoint during the peroperative phase. We have used the technique on 50 occasions. In all cases, it gave a linear vaginal scar without granuloma. Used in association with sub-peritoneal suction drainage it brings about a reduction in the number of lymphoceles and their complications.


[Lassa, Marbourg and Ebola viruses: new features of African tropical pathology. II. Epidemiology. Public health problems (author's transl)]. Lassa, Marbourg and Ebola viruses are characterised by their endemo-epidemicity in tropical Africa, by their potential of inter-human transmission, by their gravity (30 to 50% mortality in cases admitted to hospital) and by the difficulty of their aetiological diagnosis. This results in a public health problem for countries in non-endemic regions receiving travellers coming from Africa. This problem is related to the risk of importation of cases, a risk which should not be exaggerated but nor should it be underestimated. Appropriate measures may be suggested in the light of assessment of the risk: organisation of specialised hospital facilities, laboratory and coordination service.


[Controlled trial of levamisole in rheumatoid arthritis. Clinical and immunological study of 22 cases (author's transl)]. The results of a controlled trial in 22 patients indicated that levamisole, administered in a dose of 150 mg per day for 2 months, was more effective than a placebo on the clinical symptoms and signs of rheumatoid arthritis. Side effects were seen in 8 of the 12 patients treated with levamisole. They necessitated the premature interruption of treatment in three, but were never serious (no cases of agranulocytosis were seen). In the group treated with levamisole, there was a slight decrease in sedimentation rate, a significant fall in levels of the C3 fraction of serum complement and of circulating immune complexes (detected by polyethylene glycol), and a significant increase in cutaneous reaction to tuberculin and to candidin. By contrast, there were no significant variations in levels of rheumatoid factor, serum immunoglobulins, C4 fraction of complement, lymphocyte transformation indices in the presence of six dilutions of phytohaemagglutinin, nor in the percentages of T and B lymphocytes. The effectiveness of levamisole in rheumatoid arthritis would appear to be demonstrated, but its mode of action remains open to discussion, in the absence of any definite correlation between therapeutic activity and the immunostimulant effects of the drug.


RNA-RNA interactions in the binding site of protein L24 on 23S ribosomal RNA of Escherichia coli: 1. Evidence for their occurrence between widely separated sequence regions. Protein L24, from the Escherichia coli ribosome, protects a large region of 23S RNA against ribonuclease digestion. This protected RNA consists of a series of non-contiguous subfragments encompassing about 480 nucleotides at the 5' -end of 23S RNA (1). The present work demonstrates that this RNA moiety remains intact, after removal of protein L24, indicating that the subfragments are maintained together by RNA-RNA interactions. Using a urea washing procedure, the weakly bound RNA subfragments were selectively removed leaving more strongly interacting subfragments that were identified, by gel electrophoresis and oligonucleotide fingerprinting, and shown to derive from widely separated sequence regions.


Thermodynamics of a stable yeast 5.8S rRNA hairpin helix. The 5. 8S ribosomal RNA of bakers yeast contains one particularly stable hairpin helix which is isolated by partial T1 ribonuclease digestion. Thermal hyperchromism analysis of the hairpin fragment showed that it dissociates cooperatively with 18% hyperchromism, with a Tm of 83 degree C at 2.7 mM sodium ion concentration, and with a hyperchromic difference spectrum indicative of over 90% G + C content. The probable secondary structure for the fragment was used to predict a helix free energy, delta G = -16.2 kcal/mole, which was the same as that determined from the melting equilibrium. The predicted enthalpy however, was 77% of the value, delta H = -114 kcal/mole, determined from the van't Hoff relationship. The effect on these data of a G.U base pair within the 9 base pair helix is discussed.


Small-angle X-ray titration of the complex formed between the ribosomal protein S4 and its 16S binding site, S4-RNA: a central core in the 30S subunit. X-ray scattering titrations at 21 degree C and in ribosomal reconstitution buffer indicate that the S4-RNA and the protein S4 from a 1:1 complex with a stability constant, log  K approximately 6.5. When the complex forms, there is only a limited change in the scattering curve indicating that S4-RNA essentially retains its conformation during the complex formation. The increase in the gyration radius as a result of the complex formation, delta R = 4 +/- 3 A, as well as the experimental scattering curve of the complex can be explained by models where the protein S4 is supposed to interact with the periphery of the S4-RNA.


Large-scale isolation of a native deoxyribonucleohistone complex from baker's yeast. A method for large scale isolation of a native deoxyribonucleohistone complex from yeast is described. Crude chromatin, obtained after disrupting yeast cells at low ionic strength, contains a large amount of lipids, partially due to contaminating membranes. Most of them are removed by a Triton X-100 treatment, followed by step-gradient centrifugation. About 90% of the pellet may be solubilized by mild procedures, the composition of the soluble material being: histone/DNA = 1.0;nonhistone proteins/DNA = 0.55; RNA/DNA = 0.18. Histones can be obtained with high purity. Micrococcal nuclease digests DNA to yield a series of oligomeric fragments, with an average repeat length of about 160 base pairs. Circular dichroism spectra show that (theta) 270 is reduced by about 30% when compared to pure DNA and that chromosomal proteins are not denatured. These results indicate that the components of the complex conserve the native state.


A computer aided oligonucleotide analysis provides a model sequence for RNA polymerase-promoter recognition in E.coli. A novel computer procedure has been used to search for homology among 17 known procaryotic promoter sequences. A model sequence, :formula: (see text), is compatible with the properties of all known promoter and operator mutations, predicts base positions for the initiation of RNA synthesis coinciding with those determined experimentally, is compatible with current models for the regulation of transcription, suggests that RNA polymerase could recognize the DNA double helix firstly in the B conformation then in the A.


Preparation of synthetic tRNA precursors with tRNA nucleotidyltransferase. Rabbit liver tRNA nucleotidyltransferase can be used to substitute nucleotides within the -C-C-A sequence of tRNA or to add nucleotides following this sequence. These anomolous reactions of the enzyme have been used to prepare radioactively-labeled synthetic tRNA precursors which mimic the structure of the natural precursors. Under appropriate conditions synthetic precursors of defined structure can be made. In this paper we describe the synthesis of tRNA-C-[14C]U and tRNA-C-C-A-[14C]C-C, which are representative of tRNA precursors containing altered residues within the -C-C-A sequence or with extra residues following the normal 3'terminus. A variety of other possible precursors can also be prepared. These synthetic tRNA precursors have already proved useful for isolation of possible tRNA processing nucleases.


Purification of potential 3' processing nucleases using synthetic tRNA precursors. The synthetic tRNA precursors, tRNA-C-114C]U and tRNA-C-C-A-[14C]C-C, as well as poly (a) and diesterase-treated tRNA, have been used to identify and purify potential 3'processing nucleases. Four activities have been separated by this analysis; and three of them have been characterized. Two of the enzymes, which are well-separated on hydroxylapatite columns, act on poly(A), require K+ and Mg2+ for activity, and have molecular weights of about 90,000. These activities have properties previously ascribed to RNase II. The third enzyme does not act on poly(A), requires Mg2+ for activity, and has a molecular weight of about 60,000. It is identical to RNase D, previously characterized as an exonuclease acting on tRNAs with altered structure. Each of the enzymes can remove nucleotides from the tRNA precursor containing extra nucleotides beyond the 3'terminus, whereas they are relatively inactive with intact tRNA or tRNA-C-U. The greatest specificity was displayed by RNase D. The possibility that RNase D is a 3'processing nuclease is discussed.


Interaction of N-acetyl-phenylalanyl-tRNAPhe with 70S ribosomes of Escherichia coli. The interaction of N--Acetyl--Phe--tRNA Phe with 70 S ribosomes is a reversible process in the absence as well as in the presence of messenger. The equilibrium binding constants of these interactions were measured at different magnesium concentrations and temperatures and thermodynamical quantities computed. The enthalpy of the formation of complexes with the P site of ribosomes is larger by 6,000 cal/mol in the presence of poly (U) than in the presence of poly (C) or in total absence of messenger. Free energy differences are rather small, the association constants differ less than one order of magnitude. The association constant of N--Acetyl--Phe--tRNA Phe with the A site of ribosomes is 30--50 times lower than with the P site even in the presence of poly (U).


Pulsed FT-NMR double resonance studies of yeast tRNAPhe: specific nuclear Overhauser effects and reinterpretation of low temperature relaxation data. Cross-relaxation effects are demonstrated between the imino protons and other protons in yeast tRNAPhe and H2O. A detailed examination has been made of the observed relaxation rate of the proton resonance at 11.8 ppm from DSS as a function of the D2O content in the solvent. This result, as well as the size and number of observed nuclear Overhauser effects, suggests that dipolar magnetization transfer between solvent H2O, amino, imino, and other tRNA protons may dominate the relaxation processes of the imino protons at low temperature. At higher temperatures the observed relaxation rate is dominated by chemical exchange. The selective nuclear Overhauser effects are shown to be an important aid in resonance assignments. By these means we were able to identify tow protons from the wobble base pair GU4 at 11.8 ppm and 10.4 ppm.


An Escherichia coli ribonuclease which removes an extra nucleotide from a biosynthetic intermediate of bacteriophage T4 proline transfer RNA. The biosynthesis of bacteriophage T4 tRNAPro, tRNASer, and tRNAIle requires enzymatic removal of extra nucleotides from the 3' terminus of the respective precursor RNAs. A ribonuclease activity capable of catalyzing such reactions has been partially purified from uninfected Escherichia coli using an artificial precursor RNA as substrate. A number of ribonuclease activities were resolved during purification. Use of E. coli strain BN, a mutant known to be deficient in the relevant ribonuclease activity, permitted us to identify it in wild-type cells. This activity was designated the BN ribonuclease. BN ribonuclease had an apparent molecular weight of 35,000 as measured by Sephadex gel filtration. Mg2+ was required for activity, which was optimal at [Mg2+] of 2mM. Activity did not require monovalent cations K+ or Na+. BN ribonuclease was less efficient at removing extra residues in the biosynthesis of tRNASer and tRNAIle than in the biosynthesis of tRNAPro.


Isolation of Q nucleoside precursor present in tRNA of an E. coli mutant and its characterization as 7-(cyano)-7-deazaguanosine. One of the E. coli mutants selected for deficiency of modified nucleoside Q was found to contain an analogue of Q and normal guanosine in place of Q. The analogue of Q, designated as preQo, was isolated on a large scale from purified tRNATyr containing preQo. The structure of preQo was determined to be 7-(cyano)-7-deazaguanosine by comparison of its ultraviolet absorption spectra, thin-layer chromatographic mobility and mass spectrum with those of synthetic material.


T7 gene 6 exonuclease has an RNase H activity. T7 gene 6 exonuclease has been shown to have an RNase H activity as well as a double-strand specific DNase activity by the following experiments: The RNase H activity coelutes with the DNase activity from DEAE-cellulose, phosphocellulose, hydroxyapatite, and Sephadex G-200 columns. Gene 6 exonuclease specified by a T7 strain with a temperature sensitive mutation in gene 6 has an extremely heat-labile RNase H activity as well as a heat-labile DNase activity. T7 gene 6 exonuclease degrades the RNA region of a poly(A) . poly(dT) hybrid polymer exonucleolytically from the 5' terminus, releasing a ribonucleoside 5'-monophosphate product. When the RNA strand of a 0X174 RNA . DNA hybrid molecule synthesized with E. coli RNA polymerase is degraded, a ribonucleoside triphosphate is produced from the 5'-triphosphate terminus. Participation of T7 gene 6 exonuclease in the removal of primer RNA in discontinuous replication of T7 DNA is discussed.


Separation of ribosomal subunits of Escherichia coli by Sepharose chromatography using reverse salt gradient. A mixture of 30 S and 50 S subunits quantitatively absorbs on a column of Sepharose--4B from the buffer: 0.02 M Tris--HCl, pH 7.5, containing 1.5 M (NH4)2SO4. During elution by reverse gradient of ammonium sulphate (1.5--0.05 M) the subunits are eluted at different salt concentrations. Complete separation of subunits is attained in the absence of Mg2+ ions. The 30 S subunits prepared from 70 S ribosomes according to this procedure are fully active in the codon--dependent binding of a specific aminoacyl--tRNA. After their reassociation with 50 S subunits isolated by zonal centrifugation, the resulting 70 S ribosomes are active in polypeptide synthesis at the same degree as control 70 S ribosomes in which both types of subunits were prepared by zonal centrifugation. The initial 70 S ribosomes for the chromatographic separation into subunits can be obtained by their pelleting from a crude extract with subsequent washing with concentrated solutions of NH4Cl in the ultracentrifuge, or by salt fractionation of the crude extract according to a slightly modified procedure of Kurland.


Isopentenyladenosine deficient tRNA from an antisuppressor mutant of Saccharomyces cerevisiae. We have isolated a mutant of Saccharomyces cerevisiae that contains 1.5% of the normal tRNA complement of isopentenyladenosine (i6A). The mutant was characterized by the reduction in efficiency of a tyrosine inserting UAA nonsense suppressor. The chromatographic profiles of tRNATyr and tRNASer on benzoylated DEAE-cellulose are consistent with the loss of i6A by these species. Transfer RNA from the mutant exhibits 6.5% of the cytokinin biological activity expected for yeast tRNA. Transfer RNAs from the mutant that normally contain i6A accept the same levels of amino acids in vitro as the fully modified species. With the exception of i6A, the level of modified bases in unfractionated tRNA from the mutant appears to be normal. The loss of i6A apparently affects tRNA's role in protein synthesis at a step subsequent to aminoacylation.


Incorporation of labelled degradation products of radioactive thymine into non DNA material. When thymine auxotrophs are grown in the presence of methyl labelled [3H] or [14C] thymine which has been stored at 4 degrees C, two classes of material are labelled which are not DNA. One class sediments on neutral sucrose gradients with spontaneously single stranded Okazaki pieces, is unstable in alkali, migrates on alkaline gels as very small material and is digested by ribonucleases and micrococcal nuclease, but not by DNAase I. This class is presumably RNA. The second class sediments more slowly on both neutral and alkaline sucrose gradients than Okazaki pieces, but co-migrates on alkaline gels with DNA whose size is between 700 and 4000 nucleotides. It is not digested by alkali, ribonucleases or deoxyribonucleases. Its identity is unknown. The proportion of the total incorporated counts in these two classes depends on the time of storage of the thymine and is already sufficient to interfere with certain types of experiments when the thymine is only a few weeks old. Thymine is easily purified by paper chromatography and this purified thymine does not label the two non DNA classes of material. It is recommended that radioactive thymine be purified in this way before use.


Clinical application and results of the assessment of coronary blood flow by the regional precordial xenon residue detection technique. Methodological and technical aspects as well as application and results of the precordial Xenon-residue-detection technique are critically reviewed. The results concern mainly normal flow in various regions of the heart esp. in the free wall of the right and left ventricle, poststenotic flow in patients with coronary artery disease in relation to the degree of proximal narrowings as well as wall motion of the corresponding LV segment, bypassgraft flow and flow after drug interventions esp. nitrates, betablockers, the calcium-antagonist Nifedipine and the coronary dilator Dipyridamole. In spite of its serious limitations (high affinity of Xenon for fatty tissue, geometrical problems in the assessment of flow and its relation to anatomy, gas exchange in situations of high flow etc.), the technique is found to be a useful investigatory tool. Due to its technical display and the related high costs routine application is, however, prohibitive.


Histochemical identification of T and B areas in paraffin-embedded lymphoid tissues by demonstration of alpha-naphthyl acetate esterase (ANAE) activity. A strong activity of acid alpha-naphthyl acetate esterase (ANAE) in T lymphocytes and macrophages within mouse lymphoid organs is demonstrable even after paraffin-embedding, if a rapid embedding procedure with acetone as dehydrating agent is used. The lymphoid tissue structure is better preserved in paraffin sections procured after this rapid embedding procedure than in frozen sections commonly used for histochemical demonstration of ANAE. In the paraffin sections the pattern of the ANAE reaction allows a clear delineation of the T and B areas in the lymphoid organs, and the intracellular location of the focal ANAE activity in T lymphocytes is readily discernible.


[Complications following injection of fluid silicones into tissue and indications for their clinical use]. On the basis of literature and the results of investigations performed in the Department of Experimental Surgery and Plastics Research of the Institute of Surgery, Medical Academy in Wroclaw (Poland), the author discusses early and late complications after intratissular injections of silicone fluids. The indications for their clinical use are limited.


[Hydrogen peroxide, chloramine T and chlorhexidrine in the disinfection of acrylic resin]. The effectiveness of 3% h drogen peroxide, 5% chloramine T and 0,5% chlorhexidine gluconate solutions in disinfection of acrylic resine plates massively infected with oral flora was analysed. The acrylic resine plates used for investigations, were infected in vitro with mixed salivary flora characterized by small numbers of yeast-like fungi (1st group), or great number of these microorganisms (2nd group). Infected plates were exposed to solutions of analysed disinfectants during various time periods. After rinsing or inactivation of disinfectant residues, acrylic plates were put into bacteriological medium and incubated during 7 days period in 37 degrees C. The results of this study indicated the effectiveness of acrylic plates disinfection to be dependent on used disinfectant, time of exposition, and microorganisms present on the surface of acrylic resine. The solutions of disinfectants were less active in the cases of plates infected with material containing great numbers of yeast-like microorganisms. Among analysed disinfectants 0,5% solution of chlorhexidine was characterized by most effective and rapid activity, whereas 3% solution of hydrogen peroxide was found to be the least effective.


[The yesterday and today of alloplastic materials]. The authors present establishment and development of works connected with manufacturing of knitted materials for medicine in the Research and Development Centre of the Knitting Industry in Lódź. The range of products in the years of 1959-1978, the characteristic of research works and the in projects for the next years are described.


Diagnosis of treatable types of short and tall stature. Some of the most important types of small and tall stature which are accessible to treatment are discussed. The causes of small stature are multiple, since any severe chronic illness during childhood may inhibit growth and the most specifically treatable disorders (e.g. hypothyroidism and hypopituitarism) are the least frequent ones. In tall stature, the causes are less varied and genetically determined, familial tall stature is predominant. The point is stressed that the basis of an accurate diagnosis in these conditions is (1) a thorough understanding of the dynamics and variations of normal growth and (2) knowledge of methods concerning growth analysis, evaluation of bone maturation and prediction of adult height. Sophisticated laboratory procedures, such as determinations of growth hormone, are necessary in only a few selected cases.


Neuronal migration and contact guidance in the primate telencephalon. Over the last decade, evidence from experimental studies on neuronal migration in non-human primates has accumulated to the point where it can significantly amplify our understanding of the normal and pathological development of the human telencephalon. Systematic analysis of neuron genesis by the method of H3-thymidine autoradiography shows that in rhesus monkeys all neurons destined for the neocortex are generated near the surface of the lateral ventricle during a two-month period in the middle of gestation. Following their last cell division, young neurons migrate outwards across the cerebral wall to the developing cortical mantle, a journey that requires one to three days at early stages of neurogenesis, or more than two weeks towards the end of cortical development. From the very beginning, the basic columnar organization of the neuroepithelium favours radial migration. During later stages, when the primate telencephalic wall expands unevenly in thickness and surface area and begins to form primary fissures and cerebral promontoria, young neurons migrate to their cortical destinations in apposition to fascicles of radial glial fibres which span the full distance between the ventricular and pial surface. Furthermore, it appears that several generations of neurons all originate in the same restricted location at the ventricular surface, migrate along the same glial fascicles and consequently accumulate in the same radial cortical 'columns' in which, as a rule, somas of later generated neurons take positions external to the somas of their predecessors. It is proposed that fascicles of radial fibres (a) facilitate neuronal migration to the distant cortical plate through a complex assembly of closely-packed cells and processes that compose the developing primate telencephalon; (b) provide constraints which preserve a radial alignment of clonally related neurons in cortical columns; and (c) reproduce the mosaicism of the germinal ventricular zone at the expanded and curved cerebral surface.


[Aspartate aminotransferase activity of different strains of Escherichia coli]. Aspartate-aminotransferase activity was measured in strains of Escherichia coli grown no meat infusion broth. As an enzyme cells with damaged permeability of the cytoplasmatic membrane were used. The cells were damaged by an addition of toluene at a concentration of 1% of the reaction mixture by volume. These cells showed 50--60% aspartate-aminotransferase activity as compared with aliquotes of the cell-free extract. E. coli synthesized oxalacetic and glutamic acids from L-aspartic acid and alpha-ketoglutaric acid. Glutamic acid was identified by thin-layer chromatography on Fixion plates. E. coli str. 85 cells and their extracts grown on the synthetic medium containing glucose or glycerol showed a two-fold specific aspartate-aminotransferase activity as compared with those cultivated on meat-infusion broth.


[Glucagon secretion in several endocrine diseases]. The authors examined 48 patients with different endocrine pathology (relatives of patients with diabetes mellitus with a normal glucose tolerance test, patients with diabetes mellitus, obesity, thyrotoxicosis, and hypothyroidism) and a group of healthy persons. Blood glucagon concentration was determined radioimmunologically on fasting stomach and against the background of insulin hypoglycemia. A marked reduction of glucagon on fasting stomach was noted in patients with diabetes mellitus, and a reduction of the hormone concentration 30 and 60 min after the insulin injection. In obese patients and relatives of diabetic patients the initial blood glucagon level was not different from that in healthy persons. At the same time there was a significant reduction, and in relatives of diabetes patients also a retardation of glucagon secretion against the background of insulin hypoglycemia. The pattern of glucagon secretion in thyrotoxicosis and hypothyroidism proved to be changed.


Three-dimensional structures of aspartate carbamoyltransferase from Escherichia coli and of its complex with cytidine triphosphate. X-ray diffraction studies to nominal resolutions of 3.0 A for unliganded aspartate carbamolytransferase (EC 2.1.3.2)(R32 crystal symmetry) and of 2.8 A for the complex of aspartate carbamoyltransferase with cytidine triphosphate (P321 crystal symmetry) have yielded traces of the polypeptide chains of the catalytic (C) and regulatory (R) chains in the hexameric C6R6 molecules. The independent molecular structures of the liganded and unliganded forms of the enzyme are very nearly identical. In the regulatory chain there is a CTP-binding domain that interacts with an adjacent regulatory subunit and a zinc-binding domain that interacts with the catalytic subunit. In the catalytic chain a polar domain shows interactions between adjacent pairs of C chains to form each trimer C3 while an equatorial domain shows intramolecular C3--C3 interactions. The active site is at or near the interface between adjacent C chains within the trimers. Probably each active center involves amino acid residues from adjacent C chains.


A photo-CIDNP study of the interaction of oligonucleotides with gene-5 protein of bacteriophage M13. It is shown that photo-CIDNP effects (CIDNP, chemically induced dynamic nuclear polarization) can be generated in the 360-MHz proton NMR spectrum of gene-5 protein from bacteriophage M13. This technique is used to determine the number of tyrosyl residues at the surface of the protein and to assign the resonances from the 3,5-ring protons of these residues. The DNA-binding site of the protein is investigated by formation of complexes with oligonucleotides. Complex formation leads to shifting and/or quenching of the photo-CIDNP emission signals of the surface tyrosines, implying that they are involved in DNA-protein interaction. These experiments are complemented by studying the complex formation of Lys-Tyr-Lys to poly(A).


Contacts between Escherichia coli RNA polymerase and a lac operon promoter. The chemical alkylating agent dimethyl sulfate can probe the interaction between Escherichia coli RNA polymerase (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) and the purine bases of a promoter. This agent methylates the N7 position on guanine or the N3 position on adenine; the bound protein can either protect these positions or affect the reactivity to produce an enhanced methylation. The pattern of DNA residues in the lactose promoter protected from, or enhanced to, methylation by a specifically bound polymerase shows that the enzyme covers a region of at least 38 base pairs, stretching upstream from the origin of transcription. These protein-DNA contacts occur predominantly in the major groove of the DNA helix. Furthermore, this pattern of methylation shows that the polymerase unwinds the helix at the origin of transcription. The relationship between polymerase-DNA contacts defined by dimethyl sulfate and known features of promoter structure is discussed. To facilitate these experiments I have constructed a plasmid that permits a unique 5'-end labeling of each strand of a 95-base-pair fragment containing a lac operon promoter. This plasmid contains two copies of the lac promoter-operator region.


Pulvomycin, an inhibitor of protein biosynthesis preventing ternary complex formation between elongation factor Tu, GTP, and aminoacyl-tRNA. Pulvomycin and the synonymous antibiotics labilomycin and 1063-Z are shown to inhibit prokaryotic protein synthesis by acting on elongation factor Tu (EF-Tu): in the presence of the antibiotic, the affinity of EF-Tu for guanine nucleotides is altered, the EF-Tu.GDP/GTP exchange is catalyzed, and the formation of the EF-Tu.GTP complex is stimulated. Hydrolysis of GTP by EF-Tu, induced by aminoacyl-tRNA, ribosomes, and mRNA or by kirromycin, is inhibited by pulvomycin. As shown by Millipore filtration, chromatographic analysis, and hydrolysis protection experiments, pulvomycin prevents interaction between aminoacyl-tRNA and EF-Tu.GTP to yield the ternary complex aminoacyl-tRNA.EF-Tu.GTP. Thus, enzymatic binding of aminoacyl-tRNA to ribosomes is blocked.


Oxidation of the methionine residues of Escherichia coli ribosomal protein L12 decreases the protein's biological activity. Oxidation of ribosomal protein L12 with hydrogen peroxide converts the three methionine residues to methionine sulfoxide. The oxidized protein has a decreased ability to bind to ribosomes, interact with ribosomal protein L10, be precipitated by L12 antiserum, and serve as substrate for the acetylating enzyme that converts L12 to L7. Full activity of L12 is regained when the protein is reduced with 2-mercaptoethanol. Sedimentation equilibrium analysis shows that oxidation of the methionine residues in L12 causes the conversion of the protein from the dimer to the monomer form, and the results indicate that the dimer is the active form of the protein in the above reactions.


Molecular analysis of cloned human 18S ribosomal DNA segments. A fraction of DNA from the human fetal lung fibroblast line IMR-90, 30-fold enriched for ribosomal DNA, was cloned in the lambda phage vector Charon 16A. Of 978 clones assayed by hybridization to a mixture of 125I-labeled 18S and 28S ribosomal RNA, 11 recombinants containing a 3.8-megadalton segment of human 18S ribosomal DNA were identified. Restriction endonuclease analysis of these clones demonstrated variation only in orientation of the human gene segment within the phage vector. Restriction sites that we had previously detected from analysis of restriction products of unfractionated human DNA by using the Southern transfer method were also present in the cloned DNA segment. Recombinant DNA technology thus provides a valid and efficient means to define structural conservation or variation within families of human genes.


Requirement of a plasmid-encoded protein for replication in vitro of plasmid R6K. Conditions are described for the replication of exogeneous R6K DNA in an in vitro system prepared from Escherichia coli cells. Replication of plasmid DNA in this system is semiconservative and sensitive to actinomycin D, novobiocin, arabinofuranosyl-CTP,N-ethylmaleimide, and inhibitors of DNA-dependent RNA polymerase. An ammonium sulfate fraction prepared from cells carrying the R6K plasmid is required for replication. A direct role in replication for a plasmid-encoded protein, designated pi, in this fraction is indicated by the inactivity of this fraction when prepared from cells carrying a temperature-sensitive mutant plasmid and the thermolability of this fraction when prepared from cells carrying a partial revertant of the mutant plasmid. This plasmid-encoded protein is necessary for the initiation of R6K DNA replication and functions before or during the formation of nascent RNA in the initiation process. The results of titration assays of this protein using various template DNAs suggest that the protein interacts with the plasmid DNA at the region essential for DNA replication.


Construction and characterization of a 2.5-kilobase procollagen clone. Recombinant bacterial plasmids have been constructed by inserting double-stranded chicken procollagen cDNA sequences linked to chemically synthesized decanucleotides containing HindIII sites into the HindIII site of pBR322. After transformation of Escherichia coli chi1776, colonies were selected by ampicillin resistance and recombinants containing procollagen sequences were identified by colony hybridization to 32P-labeled procollagen cDNA. The inserts from three recombinant plasmids, pCg10, pCg13, and pCg45, were 1200, 2200, and 2550 base pairs long respectively. Their sequence homology has been established by restriction mapping and crosshybridization of nick-translated plasmids to Southern blots of Hpa II fragments of the inserts, pCg45 has been positively identified as containing the pro alpha2 collagen sequence by partial determination of the DNA sequence of its ends: it has a short thymine-rich sequence at one end and a sequence coding for residues 478--499 in the chicken alpha2 chain at the other end.


Structure of the lambda att sites generated by int-dependent deletions. Bacteriophage lambda integrates into the chromosome of its Escherichia coli host by means of a site-specific recombination between a locus on the phage chromosome (phage att site) and a locus on the bacterial chromosome (bacterial att site). The nucleotide sequence of four lambda att sites altered in site-specific recombination has been determined. The int-dependent deletions that generated these att sites have one end point within the phage att site and extend either to the left or to the right. As a result of the new internucleotide bond created by deletion formation, these phage have alterations in the 15-base-pair common core region. The new DNA sequences brought to the att sites by the deletions, designated delta for regions to the left and delta' for regions to the right, do not share any discernible homology with their analogous counterparts in the phage att site arms, P and P', respectively, or with the bacterial att site arms, B and B', respectively. The finding of alterations in the 15-base-pair common core region necessitates a reinterpretation of the genetic properties of these att sites in site-specific recombination. The structure of these sites in relation to their genetic properties can be viewed as being consistent with a model in which the only specificity elements in int-dependent site-specific recombination are the common core region, O, and the phage arms, P and P'.


Transcription termination: nucleotide sequence at 3' end of tryptophan operon in Escherichia coli. We have determined the RNA and DNA sequences in the region specifying termination of transcription at the end of the tryptophan (trp) operon of Escherichia coli. A 3'-terminal mRNA fragment of about 150 nucleotides yielded oligonucleotide products that could be assigned to the end of trpA (the last structural gene in the operon) by correlation with the amino acid sequence of the protein product. Analysis of the DNA corresponding to this region served to align the few noncoding RNA oligonucleotide sequences and demonstrated that termination of trp transcription occurs in vivo at a site 36 nucleotides after trpA, with greater than 95% efficiency. In two different strains partially defective in the transcription termination factor rho, the purified transcript is much longer and more complex, suggesting that a significant amount of read-through occurs in these strains. This is consistent with evidence [Guarente, L. P., Mitchell, D. H. & Beckwith, J. (1977) J. Mol. Biol. 112, 423-436] that efficient termination in vivo at the end of the trp operon is a rho-dependent event. The trp terminator (trp t) shares several features with other known sites of transcription termination, including (i) a 3'-terminal RNA sequence of several uridine residues, C-A-U-U-U-U(OH), (ii) a G.C-rich region in the DNA immediately preceding the site of termination, followed by an A.T-rich region, and (iii) a region of dyad symmetry in the DNA which, in the transcript, is capable of forming a stable hairpin containing seven G.C base pairs and one A.U base pair in its stem.


Identification of a membrane protein as a histidine transport component in Salmonella typhimurium. A component of high-affinity histidine transport in Salmonella typhimurium has been identified. It is a basic (pI about 9.0) membrane-bound protein, the P protein. It is shown to be coded for by the distal half of the previously described hisP gene by analysis of numerous hisP mutants, two of which exhibit P proteins with altered electrophoretic mobilities. Upon separation of the cytoplasmic (inner) from the outer membrane, it can be shown that the P protein is located in the cytoplasmic membrane. The P protein is under the same regulatory controls as histidine transport--i.e., transport operon promoter dhuA and nitrogen regulation. A wild-type cell contains about 200 molecules of P protein. As a result of this work we now divide the hisP gene into two genes: the hisP gene proper and the hisQ gene, which codes for another essential component of histidine transport, the Q protein. The P protein was shown previously by genetic analysis to interact with the periplasmic histidine-binding protein J, another essential component of histidine transport. Possible mechanism for the interaction of the J, P, and Q components in histidine transport, and of P and Q in lysine/arginine/ornithine transport, are discussed.


Three-dimensional electron microscopical visualization of the cytoskeleton of animal cells: immunoferritin identification of actin- and tubulin-containing structures. Cytoskeletons prepared by Triton X-100 treatment of tissue culture cells appear in stereo electron microscopy as a highly organized and interconnected three-dimensional matrix of different fibrous elements. Microfilament bundles and also tonofilament-like bundles are readily discerned when present in the cell type. In addition thinner fibers, some of which branch (smallest diameter 30--40 A), as well as fibers of larger diameter, some of which correspond to microtubules, can be seen. Since such cytoskeletons are an open, membrane-free system, individual fibrous organizations can be identified by specific antibodies. An indirect immunoferritin procedure using antibodies to tubulin or actin visualizes microtubules or actin-containing structures. Stereo electron microscopy of cytoskeletons decorated with actin antibody reveals, in addition to the F-actin-containing microfilament bundles, an extended fine actin lattice. This actin net is displayed throughout the cytoplasm not only between the microfilament bundles but also in those regions of the cytoskeleton that in the intact cell correspond to the submembraneous regions. Thus all actin-containing fibrous cytoplasmic structures may be interconnected in the living cell.


Naturally occurring promoter down mutation: nucleotide sequence of the trp promoter/operator/leader region of Shigella dysenteriae 16. The promoter/operator/leader region of the trp operon of Shigella dysenteriae 16 has single base pair differences from the corresponding region of Escherichia coli at positions -24 and -13. The difference at -13 was shown to be responsible for the 90% reduction in promoter function characteristic of the trp operon of S. dysenteriae. The base pair difference at position -13 also renders the operator partially constitutive. This allows the organism to maintain relatively high repressed levels of the trp enzymes and increases the relative importance of attenuation as a transcription control mechanism. These findings and the earlier observation that the trpE protein of S. dysenteriae is only slightly active explain the low in vivo expression of the trp operon of this organism. Nutritional studies suggest that operons involved in other amino acid biosynthetic pathways in S. dysenteriae 16 may be similarly partially inactivated.


Analysis of bacteriophage P1 immunity by using lambda-P1 recombinants constructed in vitro. We describe the dissection and reconstruction of a complex control circuit, the P1 immunity system, by a method that involves inserting EcoRI-generated fragments of P1 DNA into lambda vectors that can then be sequentially inserted into a bacterial cell. Using these techniques we have isolated lambda-P1 hybrid phages that express the products of P1 genes c1, c4, ant, and ban and, in appropriately constructed lysogens, confirmed the roles played by the first three of these products in phage immunity. In addition we have localized to particular P1 fragments the sites requisite for expression and repression of these gene products. The analysis leads to the conclusion that gpant acts in trans to antagonize repression mediated by gpc1, in support of one of two proposed models for gpant action. Moreover, two features of the immunity system are revealed: (i) a hitherto unknown component that effects gpc1 repression; and (ii) an unexpected ability of gpc4 to channel a superinfecting c1+ phage into the lysogenic state, which suggests that gpc4 activity regulates the establishment phase of lysogeny.


Evidence for translocation of DNA sequences during sea urchin embryogenesis. Hairpin-like DNA was prepared in vitro from the family of sequences that are inverted relative to each other and, as pairs, are relatively homologous and adjacent on the sea urchin genome. The majority of these hairpins are shown to have base pair mismatch positions distributed along their stems. Comparison of the hairpins derived from the DNA of morula, blastula, and gastrula stage embryos shows that during embryogenesis there are changes in the average number and position of S1 nuclease-sensitive base pair mismatch sites on the majority of the hairpin stems. Our data indicate that during early embryogenesis there are sequence changes in vivo within the majority of the adjacent inverted repeat sequences of the sea urchin genome. We have also found that there is higher specificity for the occurrence of sequence-change events within that fraction of the inverted repeat sequences that are methylated in vivo.


Protein Ia and the lamB protein can replace each other in the constitution of an active receptor for the same coliphage. Protein Ia and the lamB protein are both located in the outer membrane of Escherichia coli K-12. The lamB protein is known to be the receptor for phage lambda. Datta et al. [Datta, D. B., Arden, B. & Henning, U. (1977) J. Bacteriol. 131, 821--829] recently isolated a phage called TuIa that uses protein Ia for its adsorption. While phage TuIa fails to grow on ompB mutants, which lack protein Ia, we show here that host-range mutants of TuIa can be isolated that do grow on ompB strains. These host-range mutants fail to grow on ompB lamB double mutants, but retain the ability of the parental phage to grow on ompB+ lamB strains. They are therefore apparently able to use either protein Ia or the lamB protein for their adsorption. Genetic evidence suggests that essentially the same site on the lamB protein may be interacting with phage lambda or the host-range mutants of phage TuIa.


Immunodiagnosis of infection with Schistosoma mansoni: enzyme-linked immunosorbent assay for detection of antibody to circulating antigen. A circulating antigen, a negatively charged polysaccharide from the trematode Schistosoma mansoni, was noncovalently bound to the surface of poly(L-lysine)-coated wells in polystyrene trays, which were then used in a micro-enzyme-linked immunosorbent assay (ELISA) test. The method provides an immunodiagnostic test for schistosomiasis of exceptional sensitivity with a high degree of specificity. Comparison of Bell egg counts and ELISA titers revealed a good correlation (r congruent to 0.80) in young individuals with low to moderate worm burdens, but this relationship was less marked in older individuals or those with high egg counts.


Identification of insulin in rat brain.

Identification of insulin in rat brain. Insulin concentrations in acid/ethanol extracts of the whole rat brain were on the average 25 times higher than plasma insulin levels. Brain insulin was indistinguishable from authentic pancreatic insulin, based on its behavior in radioimmunoassay, radioreceptor assay, and bioassay and its chromatographic pattern on Sephadex G-50 column chromatography. Insulin was found in all regions of the brain examined, but distribution was uneven. Some regions had insulin concentrations as much as 100 times higher than in plasma; levels at least 10 times higher were found in other regions. The role of insulin in the central nervous system is not clear at present but, because both insulin and insulin receptors are abundant in the central nervous system, an extensive physiological regulation of the central nervous system by insulin is proposed.


Antihypertensive effect of prostacyclin (PGI2) in experimental hypertension and its influence on plasma renin activity in rats. The influence of intravenous injection of Prostacyclin (PGI2) on systemic blood pressure was investigated in conscious and anaesthetized hypertensive rats. PGI2 in doses of 1.0, 5.0 and 10.0 micrograms/kg showed a dose dependent antihypertensive effect in conscious rats with spontaneous and chronic renal hypertension. A similar response could be demonstrated in conscious rats with normal blood pressure with doses of 1.0, 10.0 and 100.0 micrograms/kg. In anaesthetized rats with acute renal hypertension or blood pressure increase, induced by continous infusion of Angiotensin II or Norepinephrine, PGI2 caused a marked decrease of blood pressure. PGI2 induced an increase of plasma renin activity in anaesthetized rats with doses of 0.1, 1.0 and 10.0 micrograms/kg. These findings support the suggestion of an antihypertensive role for PGI2 in experimental hypertension.


Prostaglandin endoperoxides promote calcium release from a platelet membrane fraction in vitro. A calcium sequestering platelet membrane fraction was prepared and the effect of arachidonic acid, PGG2 and PGH2 on calcium content evaluated. At 4 degrees C, 6.7--16.7 micrometers arachidonic acid caused significant release of calcium from preloaded vesicles. Such release was completely inhibited by aspirin pretreating the platelets from which the membrane fraction was prepared. Gamma-linolenic acid, not a substrate for prostaglandin synthesis, did not cause calcium release. At 37 degrees C, after a 5 minute calcium loading of the membrane vesicles, arachidonic acid, PGG2, and PGH2 caused release of calcium. Calcium release by the PGG2 and PGH2 was only slightly inhibited by aspirin. Imidazole, which prevented conversion of the prostaglandin endoperoxides to thromboxanes, also only slightly inhibited calcium release. Other prostaglandins including PGD2, PGE1, PGE2 and PGD2 had no effect on the calcium content of the vesicles. These studies suggest that PGG2 and PGH2 may exert their effects on platelets by mobilizing calcium from an internal membrane store to make it available to promote platelet activation.


Neuroendocrine changes in acute schizophrenia as a function of clinical state and neuroleptic medication. Changes in levels of prolactin, growth hormone, luteinizing hormone, and follicle stimulating hormone in serum, and testosterone in plasma, have been studied in 38 patients with acute schizophrenic illnesses in a 4-week double-blind comparison of the 2 isomers of flupenthixol and placebo. Only prolactin showed changes which could be related either to changes in clinical state or to the effects of medication. Prolactin levels increased during treatment with the therapeutically active alpha-isomer of flupenthixol but were unchanged with the inactive beta-isomer and placebo. Although there was a significant relationship between prolactin level and antipsychotic effect in patients on alpha-flupenthixol, in the individual case prolactin level was not a strong predictor of therapeutic response; and in patients on inactive medication changes in prolactin level could not be related to sympton change. There was a time lag of at least 2 weeks between the increase in prolactin secretion in patients on alpha-flupenthixol and the therapeutic effect attributable to medication. This delay suggests that if the antipsychotic effect is dependent upon dopamine receptor blockade it is not a direct consequence of this action. Perhaps dopamine receptor blockade permits other, and slower, changes to take place and it is these changes, rather than dopamine receptor blockade itself, which are reflected in clinical improvement.


Total anthropogenic CO2 production during the period 1800--1935 from carbon-13 measurements in tree rings. The 13C records in tree ring cellulose have been evaluated in terms of CO2 production rates on the basis of a two box model of atmosphere and ocean. We show that a linear relationship exists between the actual atmospheric excess, deltaM(t), and the actual production rate, P(t), for periods of exponential growth of the production rate. No further calibration data from other sources are needed. The results are certainly depending to some degree on the properties of the used model, but it is demonstrated that even much more complex models have essentially the same property as it has been used here. The analysis covers the perion 1800--1935, where world-wide parallel trends have been found in the tree records. The delta13C shift in atmospheric carbon due to the addition of carbon from the fossil and biospheric reservoir was -1.66% in 1935. The model analysis yields a total production of the order of 100 ppm = 212 . 10(9) tC. Roughly one half of it was produced during the last 25 years of the period. The contribution from the fossil source was 20% of the total. An open question is where the sinks are for those amounts of mobilized carbon.


Computed tomography in primary reticulum cell sarcoma of the brain. The CT findings in 16 patients with histologically proved primary central nervous system (CNS) reticulum cell sarcoma (RCS) showed this malignancy to have predilection for the basal ganglia and thalamus, the periventricular white matter, the corpus callosum, and the vermis cerebelli. In 43% of patients with untreated tumor it presented as multifocal lesions; in each of these patients the basal ganglia were involved in conjuction with another site. In the other 57% of patients with untreated tumor presenting as solitary lesions the basal ganglia, the corpus callosum, and the frontal lobe were sites of predilection. Such solitary lesions may be indistinguishable from other tumors. Contrast enhancement of RCS was characteristically homogeneous with rare lucency, even in very large lesions.


Comparison of the effects of prostacyclin (PGI2), prostaglandin E1 and D2 on platelet aggregation in different species. The activity of prostacyclin (PGI2), PGE1 or PGD2 as inhibitors of platelet aggregation in plasma from human, dog, rabbit, rat, sheep and horse was investigated. Prostacyclin was the most potent inhibitor in all species. PGD2 was a weak inhibitor in dog, rabbit and rat plasma whereas PGE1 and prostacyclin were highly active. Theophylline or dipyridamole potentiated the inhibition of human platelet aggregation by prostacyclin, PGE1 or PGD2. Compound N-0164 abolished the inhibition by PGD2 of human platelet aggregation but did not inhibit the effects of PGE1 or prostacyclin. The results suggest that prostacyclin and PGE1 act on similar sites on platelets which are distinct from those for PGD2.


Stability of authentic PGI2 during routine extraction. Clarification of the nature and sequence of products formed by the rat stomach including the origin of the ozonolysis products reported in 1971. Authentic PGI2 and PGI2 formed by rat stomach homogenates were carried through a simple extraction and purification procedure to explore the feasibility of isolation of this biologically active bicyclic ether product of arachidonic acid. The integrity of PGI2 was followed throughout by bioassay on the rat blood pressure. In this system we recently reported that PGI2 has very potent hypotensive actions which are easily distinguishable from those observed for PGE2 (14). Our results indicate that PGI2 survives the initial extraction steps (i.e. ethanol extraction, diethyl ether - HCl extraction and methylation) up to the step involving thin layer chromatography with an acidic developing solvent system. This latter procedure converts PGI2 entirely into a stable derivative, 6-keto-PGF1alpha (3,8--10). Oxidative ozonolysis of the methyl ester acetate derivative of authentic 6-keto PGF1alpha reveals products identical to those reported by Pace-Asciak and Wolfe in 1971 (1) which are also produced from authentic PGI2. This data sheds new light into 1) the nature of the biological product formed by stomach homogenates, 2) its transformation into 6-keto PGF1alpha during purification and 3) the origin of the ozonolysis products in the experiments reported in 1971.


Effect of in vivo prostaglandin treatment on 3H-PGF2alpha uptake in hamster corpora lutea. Pregnant hamsters were administered (SC) prostaglandin or vehicle on the morning of the 4th day of pregnancy. Serum progesterone was significantly depressed (p less than .01) at 0.5, 2, and 6 hours after treatment with 100 microgram PGF2alpha. Serum progesterone levels were unchanged 2 hours and 6 hours after treatment with 100 microgram PGF2beta and 2 hours after treatment with 1 mg PGF2beta. Progesterone levels were depressed to less than 1 ng/ml 6 hours after treatment with 1 mg PGF2beta. The specific uptake of 3H-PGF2alpha in whole hamster corpora lutea was significantly depressed 2 hours and 6 hours following 100 microgram PGF2alpha treatment. A 15% depression in specific uptake occurred 0.5 hour post-treatment. Treatment with 100 microgram PGF2beta resulted in no change. Administration of 1 mg PGF2beta resulted in depressed 3H-PGF2alpha uptake at both 2 and 6 hours post-treatment. Prostacyclin (PGI2) treatment resulted in no change in either 3H-PGF2alpha specific uptake or serum progesterone 2 hours after 100 microgram treatment SC. These parameters were both reduced approximately 30% 6 hours post-treatment. Treatment with 6-keto-PGF1alpha resulted in a complete lack of measurable 3H-PGF2alpha uptake and serum progesterone levels less than 1 ng/ml at both 2 and 6 hours after treatment with 1 mg SC.


Increased production of PGI2-like activity by frozen-thawed rat aorta. The effects of storage conditions, temperature, and time on the ability of the rat thoracic aorta to produce a platelet aggregation inhibitor were investigated. Aortic fragments were incubated in Tris buffer, aliquots of which were then tested for their ability to inhibit ADP-induced human platelet aggregation. The incubation fluid of samples that had been soaked in Tris buffer at 4 degrees C for 24 hours contained no inhibitor activity, whereas the incubation fluid of similar samples that had been kept at 4 degrees C but not soaked in buffer contained comparable inhibitor activity as that of fresh samples. The incubation fluid of samples that had been kept at -20 degrees C or -80 degrees C contained greater inhibitor activity than that of fresh samples, and was maintained in -20 degrees C samples for 7 days, and -80 degrees samples for 28 days. The aortic inhibitor had similar properties as PGI2.


The effect of PGI2 on canine renal function and hemodynamics. The effect of prostaglandin I2 (prostacyclin) on renal and intrarenal hemodynamics and function was studied in mongrel dogs to elucidate the role of this novel prostaglandin in renal physiology. Starting at a dose of 10(-8) g/kg/min, PGI2 decreased renal vascular resistance and redistributed the blood flow away from the outer cortex (zone 1) and towards the juxtamedullary cortex (zone 4). At 3 X 10(-8) g/kg/min, the renal vascular resistance decreased even further, but at this dose the mean arterial blood pressure also declined 13% indicating recirculation of this prostaglandin. PGI2 infusion at a vasodilatory dose resulted in natriuresis and kaliuresis. With a decline in filtration fraction, these changes were most likely secondary to the hemodynamic effects of this prostaglandin. Unlike PGE2, PGI2 had no direct effect on free water clearance indicating lack of activity at the collecting duct. PGI2 may be the important renal prostaglandin involved in modulating renal vascular resistance and intrarenal hemodynamics as well as influencing systemic blood pressure.


Localization and in vitro synthesis of prostaglandins in components of rabbit preovulatory graafian follicles. Graafian follicles obtained 9 hours after the injection of human chorionic gonadotropin (hCG) into mature rabbits were dissected into a follicular fluid component, a granulosa cell-oocyte component, and a residual wall component, (the latter containing mostly theca tissue with a small and variable amount of adhering granulosa cells). The amounts of PGE and PGF were determined for each component. The follicular fluid contained approximately 4-10 times more PGE and PGF than either the granulosa cell-oocyte component or the residual wall component. The latter two components contained approximately equal amounts of these prostaglandins. The in vitro biosynthesis of PGE and PGF was also studied and it was found that the granulosa cell-oocyte component had about 4 fold the capacity of the residual wall, and that the follicular fluid synthesized no prostaglandins. There was no significant effect of LH on either PGE or PGF synthesis in any of the components.


Formation of prostacyclin (PGI2) by the ductus arteriosus of fetal lambs at different stages of gestation. Prostaglandins appear to play a role in maintaining patency of the ductus arteriosus during gestation. Prostacyclin (PGI2) is the major product of prostaglandin biosynthesis in the lamb ductus arteriosus. This factor is both a vasodilator and a potent inhibitor of human platelet aggregation. We used inhibition of platelet aggregation as a sensitive bioassay to measure PGI2 generation in rings of ductus arteriosus from fetal lambs. Mechanical manipulation accelerated the rate of PGI2 released from the tissue 10 to 50 times. Tranylcypromine, an antagonist of prostacyclin synthetase, suppressed production of PGI2 by rings of ductus arteriosus. Rings from immature animals (98-103 days gestation, term is 150 days) released significantly more PGI2 (190 +/- 28 ng/g wet weight/ 20 min, n = 9) than did those from near term animals (136-146 days; 106 +/- 23 ng/g wet weight/20 min, n = 10). The capacity of the ductus arteriosus to generate more PGI2 earlier in gestation is consistent with the observation that vessels from animals less than 110 days gestation have a significantly larger indomethacin induced contraction than do vessels near term.


[Effect of glucose concentration on the biosynthesis of prodigiosin by serratia marcescens (author's transl)]. Serratia marcescens is an enterobacteria which produces a characteristic red pigment denominated prodigiosin. To study the effect of glucose on the kinetics of this secondary metabolite, cultures of Serratia marcescens S10 were incubated at 30 degrees C in the mineral medium GL, with glucose (2 g/l) as the carbon source. Prodigiosin production in relation to glucose consumption is studied, and parallel-wise, the effect of various concentrations of glucose on prodigiosin production. The kinetics data show the close correlation between glucose consumption and the synthesis of prodigiosin. This substrate inhibits the synthesis of pigment in cultures grown on solid medium GL with concentrations of glucose up to 15 g/l.


Effect of prolonged fasting upon insulin and glucagon secretion from isolated rat pancreatic islets. To study insulin-glucagon interrelationships in the regulation of pancreatic islet functions, glucose-mediated insulin and glucagon secretion have been studied in isolated pancreatic islets from fed and from 4 and 8-day fasted rats. At low glucose levels (50 mg %) a continuous decrease of insulin and increase of glucagon secretion were observed during prolonged fasting. High glucose concentrations 300 mg %) stimulated insulin and inhibited glucagon secretion until 4 days, but did not cause any effect after 8 days fasting. These results suggest that the secretory mechanisms of the two hormones may have a common basis.


Effect of arachidonic acid (ARA), prostaglandins E2 (PGE2), and F2alpha (PGF2alpha) on sensitized spleen cells in the hemolytic plaque assay. Spleen cells from the BDF1 generation of DBA male X C57Bl/6 female matings sensitized with sheep erythrocytes (SRBC) were exposed to various concentrations of ARA, PGE2, and PGF2alpha for 90 minutes in vitro at 37 degrees C. Following exposure these sensitized spleen cells were then examined in the hemolytic plaque assay. No changes were demonstrated in the number of plaque forming cells (PFC) with 0.1 microgram/ml of ARA treatment. However, an increase in the number of PFC occurred with increasing concentrations of ARA. A significant increase (p less than 0.01) in PFC (59%) was shown with 10.0 microgram/ml of ARA. PGE2 treatment at 10.0 and 100.0 microgram/ml showed no evidence of increase, but a slight decrease (10%) was shown with 100.0 microgram/ml. On the other hand, exposure to PGF2alpha showed a significant increase (p less than 0.05) in PFC at the 10.0 microgram/ml (35%) and the 100.0 microgram/ml (29% levels, respectively. The data suggest that both ARA and PGF2alpha enhanced PFC formation.


Elimination and organ distribution of intravenously administered allogeneic and xenogeneic IgG modifications. (Standard IgG, F (ab)2-fragments and beta-propiolactone treated IgG) in dogs. Dog IgG was produced by fractionation procedures used for the production of clinically used i.v. gammaglobulins. Chemical modification of dog IgG was done by pepsin or beta-propiolactone treatment. The intravascular half-life of beta-propiolactone IgG was 8.5 +/- 2.1 days compared to 4.5 +/- 1.6 days of pepsin treated IgG. Tissue concentrations of radioactive labelled beta-propiolactone IgG were generally higher than of pepsin digested IgG. Pepsin treated Igg was degraded to a significantly higher extent (26% of the administered radioactivity was bound to fragments smaller than 6000 MW after three days) than beta-propiolactone IgG (9% fragments after the same interval, P less than 0.001). It is concluded that the short intravascular half-life of pepsin IgG cannot be explained by increased extravascular filling, but is due to rapid degradation and excretion via the kidneys. There was no obvious difference in elimination and organ distribution between standard and beta-propiolactone IgG.


Metabolism of hydrogenated palatinose, an equimolar mixture of alpha-D-glucopyranosido-1,6-sorbitol and alpha-D-glucopyranosido-1,6-mannitol. Hydrogenated palatinose, an equimolar mixture of alpha-D-glucopyranosido-1,6-sorbitol and alpha-D-glucopyranosido-1,6-mannitol, was investigated as a potential oral sugar substitute in the following experiments in man and rat. 1. Enzymatic cleavage occurred at slow rates by maltase (alpha-glucosidase) of jejunal mucosa, liver lysosomes and yeast. 2. Part of ingested hydrogenated palatinose arrived unsplit at the caecum of the rat and underwent fermentation there; excretion in feces and urine are neglegible in man and rat. 3. Growth and maintenance of rats demonstrated 20--40 percent diminished caloric utilisation of diets containing 34.5 percent hydrogenated palatinose whereas indirect calorimetry in man showed about 50 percent caloric deficit. 4. Blood sugar did not increase in man after oral doses up to 100 g. 5. The capacity of the rat kidneys for excretion of hydrogenated palatinose and its constituents was high, symptoms of incompatibility were not observed.


Experimental Salmonella typhimurium infection in calves. The paper describes the clinical, bacteriological and pathological findings in experimental Salmonella typhimurium infection in calves. Oral doses of 10(8) and 10(9) organisms produced clinical disease and high mortality; doses ranging from 10(4)--10(7) organisms were less consistent in their action. Jersey calves appeared more susceptible to infection than Friesian calves. The clinical signs in most calves were pyrexia and a characteristic diarrhoea that lasted for up to 11 days; more severe symptoms were seen in the calves that received the higher doses. Following infection, all calves excreted S typhimurium in their faeces, the highest counts being observed in the calves that died. In the calves that survived, counts ranging from 10(2)--10(5)/g faeces occurred continuously for up to a maximum of 20 days and subsequent intermittent excretion occurred in a number of calves. In the calves that died, necrotic enteritis in the ileum and large intestine was the most striking lesion; lesions were uncommon in other organs. The findings are discussed in relation to the pathogenesis, diagnosis and control of the disease.


The quantitative assessment of normal canine small intestinal mucosa. Quanitative methods of assessing the architecture of small intestinal mucosa have been applied to biopsy material from normal dogs. Mucosal samples taken from four predetermined sites show that there are significant quantitative differences between the various levels of the small bowel. Animals of one year of age and older show no correlation between age or weight and mucosal dimensions. The significance of these findings, in relation to examination of biopsy material from cases of clinical small intestinal disease, is discussed.


The quantitative assessment of mucosa in canine small intestinal malabsorption. Histological sections from the small intestines of dogs showing clinical signs of small intestinal malabsorption have been examined using morphometric techniques. Weibel graticule measurements appear to be a useful adjunct to the subjective assessment of villus atrophy. Severe clinical signs of small intestinal malabsorption may be associated with a patchy distribution of lesions throughout the small bowel.


In vivo studies on suppressor lymphocyte activity following administration of Escherichia coli O138 lipopolysaccharide. Evaluation of Escherichia coli vaccines used in veterinary practice usually relates to antibody formation and neglects the essential characteristics of cell function leading to this end. This paper attempts to investigate the cellular responses to E coli O138 lipopolysaccharide (LPS) administered subcutaneously in a dose range of 1 microgram to 200 microgram. Rosette-forming cell responses in the draining lymph node varied from antigenic at 10 microgram to tolerogenic at 200 microgram. The tolerogenic dose also caused a marked mitogenic response as can be seen by a fourfold increase in total lymph node cells. Fractionation of normal or LPS-responding, or LPS-tolerised lymph node cells into B cell-rich and T cell-rich fractions was carried out and these were adoptively transferred simultaneously with an antigenic dose of sheep erythrocytes (SRBC) into syngeneic recipient mice. Suppressor activity of the anti-SRBC response was found after transfer of 1 X 10(6) normal B cell fractions, 1 X 10(6) tolerised B cell fractions and 1 X 10(6) tolerised T cell fractions. LPS-antigenically stimulated lymph node cell fractions had no suppressor effect but when given with Freund's complete adjuvant ensuing T cell-rich fractions produced immunosuppression in recipient mice. The significance of these findings is discussed in relation to polyclonal B cell activation, helper and suppressor T cells, and possible feedback mechanisms between interacting subpopulations of lymph node cells.


An immunisation model for the control of infectious enteritis. An intestinal immunisation procedure involving systemic priming and oral boosting was investigated in lambs with a view to providing vaccination control of enteritis in young animals. The procedure stimulated the appearance of antibody-containing cells of IgA specificity in the intestine and lambs immunised in this way with a bacterial vaccine were protected against subsequent challenge with live enteropathogenic bacteria. This immunisation regime therefore provides a useful method for the stimulation of IgA immunity in the intestine of ruminants and it is anticipated that it will have general application to a variety of enteric diseases.


Tolerance breaking by lymphocyte activating factor induced by Escherichia coli O138 lipopolysaccharide from homologous macrophages. Following an antigenic dose of 10 microgram or a tolerogenic dose of 200 microgram of Escherichia coli O138 lipopolysaccharide (LPS), BALB/c mice were examined on day 14 for percentages of theta-bearing cells. A considerable increase in T cells was noticed in lymphocytes from tolerant draining lymph nodes, and furthermore these cells did not possess receptors for LPS when tested for rosette inhibition. However, when the supernatant from 1 X 10(7) macrophages, pretreated with 150 microgram LPS, was given to tolerant mice on day 7, by day 14 tolerance was found to be broken, anti-LPS IgG was present in circulation and the draining lymph node contained T cells specifically committed to LPS. The change from suppressor to helper T cell activity is discussed in relation to enhancement of the immune response.


Antibody response in cattle and rabbits to early antigens of malignant catarrhal fever virus in cultured cells. Cytosine arabinoside (Ara-C) inhibited the production of late antigens and of infectious virus in monolayers of bovine kidney cells infected with the high-passage, WC-11 strain of malignant catarrhal fever virus. Early antigens were not affected. Using hyperimmune and acute-phase sera from cattle and rabbits in indirect immunofluorescence tests, it was shown that Ara-C treated cultures contained two early antigens; one was diffuse and distributed throughout the cells, the other was particulate and intranuclear. Antibody to early antigens developed later and attained lower titres in infected animals, especially rabbits; only hyperimmune sera reacted with the diffuse early antigen.


[Creutzfeldt Jakob disease. Recommended precautions for patient management and diagnostic procedures (author's transl)]. The virus of Creutzfeldt Jakob disease differs from conventional viruses in its extraordinary resistance to commonly used physical and chemical methods of decontamination. Autoclaving the virus for 1 hour at 121 degrees C and 2 atmospheres pressure is the most certain means for achieving total inactivation. Exposure to 5 p. 100 hypochlorite for several hours can also totally inactivate the virus, and exposure to phenol or potassium permanganate is probably effective, but as yet insufficiently tested.


[Contribution of stereotactic methods to diagnosis and treatment of tumors of the pineal region (author's transl)]. The respective role of surgical ablation versus palliative methods (i.e. shunting followed by irradiation) in the treatment of tumors of pineal region is still questionnable and must take into account the variability of their pathology. Thus, knowledge of tumoral type in each case is the first step to therapeutic discussion. The stereotactic methods allow a faithful biopsy, easy to interpret and without major risks.


[A study of the use of sequential chemotherapy in 176 cases of glioblastoma (author's transl)]. Sequential administration of VM 26 and CCNU has been used in France for the treatment of glioblastoma since 1973. Results have shown that this association can cause a remission lasting, on average, from seven to eight months after starting chemotherapy. It seems that, adding Adriamycin to this sequential therapy does not increase its efficiency. On the other side, dosage and the interval between cycles of chemotherapy appear to be a determining factor in the activity, but the limits are very narrow. Therapy is either ineffective or has doubtful efficiency in about one third of patients, and even when prolonged clinical remission is obtained, cessation of therapy, usually due to a persistent thrombopenia is followed by progression of the tumour in the following months. The large amount of research being made in the field of cellular kinetics and pharcacology will, hopefully, lead to improvement in results.


Drug interactions in rheumatoid disease--are they of any clinical importance? Much of the literature on drug interactions over-emphasizes their importance. A review of the mechanisms of drug interactions and of the clinically important interactions involving drugs commonly used in rheumatology reveals that serious interactions are likely to be uncommon. A practical approach to this problem may be divided into three parts: 1. Be aware of the mechanisms of drug interactions as they may explain some unexpected finding occurring during drug treatment. 2. Remember that serious drug interactions are more likely to involve a drug which possesses a narrow therapeutic range (e.g. oral anticoagulants). 3. When indulging in polypharmacy attempt to ensure that such treatment has real benefits. This last point may be the most important for if it is proven that combined therapy has no advantages then the potential for drug interactions will be much reduced.


The selection of patients for controlled trials. The selection of patients for testing drugs is a very complicated process even overtly; subconscious bias must be avoided and our social conscience respected. It is of prime importance to define in advance exactly what you want to test and this may include not only the therapeutic and pharmacological effects and side-effects, the long-term neoplastic potential, the mutagenic effects and the effects on the foetus, but also mundane effects like credibility, acceptance, and even taste! Patient selection is not just "Attention there Volunteers nos. 1,3,5,7,9,....step forward smartly from the left". It is a complicated process in which there is inevitably some bias and our job is to see that any biases which are included do not bias the results, and that the results are understood to apply to this particular selection of patients and not to any other. Too many explicit exclusions therefore make results less useful because less widely relevant.


Prostaglandins and lymphokines in inflammation. It is suggested that the inflammatory process in rheumatoid arthritis (RA) results from activation of small lymphocytes to produce lymphokines, whose biological properties are appropriate to a mediator of chronic inflammation. Since prostaglandins (PGs) and allied products of arachidonic acid metabolism have properties pertinent to chronic inflammation, these compounds can also be regarded as potential mediators of chronic inflammation, a viewpoint supported by the capacity of steroids and non-steroidal anti-inflammatory drugs (NSAIDs) to suppress PG formation. Certain observations are inconsistent with PGs fulfilling this role. These anomalies may be resolved by considering PGE2 formation by macrophages to be a device for regulating lymphocyte activation. A defect of lymphocyte reactivity to PGE2 provides a basis for chronicity and has been demonstrated in multiple sclerosis (MS). Alternatively, should lymphocytes in lesions of RA be susceptible to PGE2 inhibition, one consequence of NSAID treatment may be exacerbation of joint destruction.


Defective phagocytosis by synovial fluid and blood polymorphonuclear leucocytes in patients with rheumatoid arthritis. I. The nature of the defect. In a series of thirty-seven patients with rheumatoid arthritis (RA), 50% showed a defect of phagocytosis of Candida albicans by synovial fluid polymorphonuclear leucocytes (SF-PMN) and 40% yielded a defect in peripheral blood (PB-PMN). There was a positive correlation between the defective phagocytosis of SF-PMN and PB-PMN suggesting that defective PB-PMN migrate into the synovial fluid. The defect was associated with the lack of a functional C3b receptor on SF-PMN in 64% of patients and on PB-PMN in 40% of patients. SF-PMN or PB-PMN contained intracellular IgG, IgM and C3 in some patients but the presence of these factors could not be correlated with defective phagocytosis of either cell population. There was no correlation between phagocytosis and the differential agglutination test (DAT) ratio of either serum or synovial fluid. The killing of Candida by SF-PMN and BP-PMN was normal. The results could explain the increased susceptibility to joint infection of patients with RA.


Effects of drugs on prostaglandin synthesis. The role of prostaglandins in inflammatory processes is now recognized to be more complex than that of mere mediators or modulators of the acute inflammatory response. As well as their obvious vasodilator properties they have profound regulatory effects on cell function, and these may be anti-inflammatory as well as pro-inflammatory. Consequently the use of those effective non-steroid anti-inflammatory drugs which are potent prostaglandin synthesis inhibitors may in fact have detrimental effects in inflammation. Further development of cyclo-oxygenase inhibitors is less likely to lead to genuine therapeutic advance than is development of anti-inflammatory drugs with actions on other aspects of the inflammatory and destructive processes in rheumatoid disease.


Drug therapy and circulating immune complexes in rheumatoid arthritis. Circulating immune complexes have been shown to be common in rheumatoid disease and to relate to severe systemic manifestations of disease. Hypocomplementaemia and complexes detected by anticomplementary activity are found in systemic rheumatoid vasculitis and in other complications such as Felty's syndrome. By contrast elevated complement levels and complexes detected by platelet aggregating activity are common in early and in continuing active rheumatoid disease. The common presence of circulating immune complexes suggests some therapeutic manoeuvres which might be helpful in systemic rheumatoid disease. Direct attempts to alter the levels of complexes by plasmapheresis, an effective treatment for acute exacerbations of systemic lupus erythematosus (SLE), have been disappointing in rheumatoid disease. Treatment with penicillamine and with cyclophosphamide may be effective in systemic manifestations and alter the levels of circulating immune complexes. An interesting paradox exists with penicillamine which may also induce a nephropathy with all the features of an immune complex deposition disease. The mechanism may relate to an alteration in the type of immune complex circulating, with a loss of the "protective" effect of rheumatoid factor.


Inflammation in osteoarthritis.

Inflammation in osteoarthritis. Osteoarthritis is usually regarded as a non-inflammatory disease. However, several authors have stressed the importance of an inflammatory phase early in the development of the condition, and specific inflammatory types of osteoarthritis have been described. Comparative trials of pure analgesic and anti-inflammatory drugs in a small sub-group of these patients showed a similar reduction of pain with both types of agent. However, a significantly greater improvement in joint tenderness and stiffness was found during treatment with anti-inflammatory drugs.


A comparison of ketoprofen and naproxen in rheumatoid arthritis. Ketoprofen, 50+50+100 mg, was compared to naproxen, 250+250+250 mg, in a double-blind, cross-over twice 4-weeks' study on patients with RA. There was no significant difference in the effect on morning stiffness, pain at rest, joint count, grip strength or ESR. Among 28 patients 10 preferred ketoprofen and 7 naproxen. Two ketoprofen and 1 naproxen periods were interrupted owing to intolerable side-effects. Twenty of the patients experienced some side-effect from at least one drug. The most common complaints were gastrointestinal, seen in 12 patients on ketoprofen and 9 on naproxen. Most side-effects were mild. No abnormality in blood morphology, liver function tests, serum creatinine or fasting blood glucose was observed. No occult bleeding was detected on routine stool examination.


Ketoprofen: double-blind cross-over study with indomethacin administered as a combined suppository/capsule regime in patients with rheumatoid arthritis. A preliminary report. A double-blind cross-over study is being undertaken in fifty patients with "definite" rheumatoid arthritis, comparing ketoprofen (2-(3-phenylbenzoyl proprionic acid) 75 mg daily orally and 100 mg suppository at night with indomethacin in the same dosage. A pilot study of twenty-four patients has been completed, fifteen having finished the trial. The objective assessment of grip strength and articular index were comparable with the two drugs, but subjective assessment showed that relief of early morning stiffness was better with indomethacin, although pain relief was no different. Evidence so far available shows that ketoprofen is a useful anti-inflammatory analgesic drug which is well tolerated and that the use of a suppository may be helpful in a few patients who find the oral route unacceptable. A larger study is necessary to evaluate finally the use of a combined administration.


Possible causes of treatment failure with the NSAID. The clinical efficacy of non-steroidal anti-inflammatory drugs (NSAID) is often disappointing in spite of their well-known antiphlogistic actions. The reasons for this failure are probably due to irregular intake by patients on long-term treatment with such agents. These considerations inevitably point to the necessity of concomitant determination of the plasma levels of all NSAID tested, and the need to perform clinical trials in an appropriate setting in which such determinations can be conveniently carried out.


Perforations of the nasal septum following operative procedures. New operative methods have been introduced into nasal surgery in recent years; these are septoplastic, rhinoplastic and nasal reconstruction. These new methods are now employed together with the more traditional submucous septum resection. Today the above mentioned operations have been carried out in such large numbers that it is possible to commence comparing operative results. The present investigation is a comparison of the number of septum perforations with 1) submucous septum resection, 2) septum correction with reconstruction of the septum skeleton, and 3) septum correction in connection with rhinoplastic of the outer nose, possibly supplemented by nasal reconstruction. In the latter case a systematic suturing of all mucosal lacerations has been carried out, partly to prevent septum perforations and partly in order to keep the septum flaps intact during reimplantation of the cartilage and bone with the object of constructing a solid septum skeleton. While primary suturing of the mucosa is a relatively simple procedure, the closure of postoperative septum perforations is far more difficult. The results depend on the size of the perforation and the amount of cartilage and bone material still remaining in the septum. It must therefore be advisable to employ operative techniques which are able to prevent septum perforations or at least reduce their number to a minimum.


[Fixation by longitudinal pinning of two rib grafts in a case of loss by substance of the horizontal rami of the mandible (author's transl)]. In a difficult case of bilateral loss of substance of the body of the mandible, with aplasia of the residual tissue, in an elderly psychotic patient, reconstruction was obtained simply by the insertion of two rib grafts each fixed by a single Kirschner's wire (removed four months later).


[Several cases of emergency repair of loss of skin substance in the child (author's transl)]. Traumatic loss of skin substance in the child is uncommon. There are three main causes: dog bites, scraping lesions and bullet wounds. In all cases, great attention is paid to initial and early treatment of the wounds (exploration, wound care) under general anaesthesia. Provided the operator is competent, dog bites may be completely repaired from the outset. Lesions due to scraping or bullet wounds should be treated more cautiously with emergency treatment not necessarily seeking to achieve closure at any price.


[Reconstruction of the lower lip for sequelae of noma (author's transl)]. The authors present a case of reconstruction of the height of the lower lip for sequelae of probable noma in a 35-year-old Algerian. They used a bipediculated Dufourmentel flap. The morphological and functional result was satisfactory.


The effect of parietal cell vagotomy and selective vagotomy with pyloroplasty on gastric acid secretion. A prospective randomized study. The insulin- and pentagastrin-stimulated gastric acid secretion was studied before and 1 week, 1 month, 6 months and 1 year after parietal cell vagotomy (PCV) and selective vagotomy with pyloroplasty (SV + P) in 50 patients who took part in a consecutive randomized trial. The randomization and the operations were performed in such a way that the extent of the denervation of the proximal stomach was identical in the two groups. Basal acid output and the insulin-stimulated acid secretion were significantly increased from 1 month to 1 year after PCV, while there was no such increase after SV + P. There was no change of the pentagastrin-stimulated acid secretion (PAOP) from 6 to 12 months in either groups. One year postoperatively PAOP was significantly greater after PCV than after SV + P. The results are compatible with vagal reinnervation after PCV and support the assumption of a sprouting from the innervated gastric antrum.


A comparison of oral prednisolone given as single or multiple daily doses for active proctocolitis. Forty mg has been shown to be the optimal daily dosage of prednisone for outpatients requiring oral steroids for active proctocolitis. Although daily doses of oral steroids are commonly divided, a single dose each morning causes less adrenal suppression and is more convenient to take. A randomized controlled trial has been performed on patients with proctocolitis, in which 23 received 40 mg prednisolone each morning as one dose, and 22 received 10 mg four times a day, over two weeks. Physicians unaware of the dosage schedule scored the disease activity and assessed the steroid side-effects when the patient entered the trial, at day 7 and at day 14. Of those taking the divided dose the disease improved in 17 and failed to improve in five. No side-effects were observed in ten patients. Of those receiving a once daily regimen, 17 improved and six did not. Nine patients had no side-effects. Further assessment showed no difference between the two groups either in response rate or side-effects produced. When oral steroids are indicated for active proctocolitis, 40 mg prednisolone, as a single dose each morning can be recommended.


Anti-fibrinolytic therapy of giant hypertrophic gastritis (Menetrier's disease). In five cases of giant hypertrophic gastritis (Menetrier's disease) biopsied gastric mucosa was examined for fibrinolytic activity; in all cases there was marked elevation of the activity due mainly to tissue plasminogen activator. The patients were given antifibrinolytic therapy with oral tranexamic acid (trans-4-aminomethyl cyclohexane carboxylic acid; trans-AMCHA), and four of the patients showed marked improvement of their hypoproteinemia as well as their mucosal disorders. One patient, who showed moderate increase of serum protein level but no reduction of the mucosal disorder, finally received gastrectomy. It was concluded that antifibrinolytic therapy seemed to block the vicious circle of 'membrane disorders', 'increased tissue fibrinolysis', 'increased vascular permeability' and 'hypoproteinemia' in Menetrier's disease.


Clonal priming of human lymphocytes with soluble microbial antigens: high-dose paralysis, restoration, and autologous leukocyte preference. Lymphocytes stimulated in appropriate leukocyte cultures undergo blastogenesis and proliferation for a finite period of time. With specific antigens the proliferative response peaks usually between 4 and 8 days, after which the blastoid cells revert to small lymphocytes. Lymphocytes "primed" in this manner can be restimulated to proliferate only by the same antigen with which they were incubated and only with an adequate amount of a self-specific, autologous, somatic product(s). First or "primary" leukocyte cultures stimulated by optimal or supraoptimal concentrations of soluble protein antigens (purified protein derivative (PPD), tetanus toxoid, Candida albicans) will undergo proliferation in the first culture, but the increased number of small lymphocytes that can be visualized after 10--14 days often fail to respond to any stimulus in second (secondary) or "primed" cultures. However, when fresh X-irradiated autologous cells are re-added in appropriate amounts, vigorous accelerated proliferation takes place. Addition of allogeneic cells to antigen-primed populations has one of three effects: (1) no effect (complete restriction); (2) in some instances allogeneic cells restore a significant response to the specific antigen but almost never to the same degree as autologous cells; and (3) allogeneic cells can also induce high levels of accelerated responsiveness without added antigen. These findings are discussed in the context of a working hypothesis that self-specific factors are involved in all specific immune responses. The combination of antigen and self-specific factors may lead to a quantitatively unique immune response to all antigens in each individual. The preferential response to antigen in conjunction with autologous rather than allogeneic leukocytes suggests that self-specific products are required for recognition of soluble microbial antigens.


Optimal conditions for polyclonal antibody secretion and DNA synthesis in human blood and spleen lymphocytes by lipopolysaccharide. Lipopolysaccharides from E. coli (LPS) stimulated increased DNA synthesis and high-rate antibody secretion in human lymphocytes from blood and spleen. Peak DNA synthesis by 10 or 100 microgram/ml LPS was seen on day 6 for blood lymphocytes. Using 500 microgram/ml of LPS the peak response was reached on day 5 and with 0.1--1 microgram/ml the maximal stimulation was on day 7. For spleen cells LPS 100 microgram/ml gave a peak stimulation on day 4 or 5. Antibody secretion, measured as plaque-forming cells (PFC) against fluorescein isothiocyanate-coupled sheep erythrocytes in a haemolysis-in-gel assay, was maximal on day 5 in blood lymphocytes and on day 3 or 4 in spleen cells. The highest cell viability and antibody secretion after stimulation with LPS was obtained in 16 mm diameter multiwell plates usng 2.5 x 10(6) cells in 1 ml of medium containing 10% AB serum absorbed with SRBC. Feeding of cultures was essential for stimulation of blood lymphocytes but not for spleen cells.


Rectal colonization with group B streptococci: relation to urogenital carriage. Urethral, cervical and rectal specimens were taken from 92 females and urethral and rectal swabs from 46 males attending a venereological outpatient department. The frequencies of group B streptococci were, in the women: urethra 24%, cervix 14% and rectum 17%; in the males: urethra 22% and rectum 7%. These findings, supported by the results of serotyping, clearly demonstrate that the human urethra is an important site of colonization and do not support the idea that the gastrointestinal canal is the primary site of acquisition.


Treatment of urgency by instillation of emepronium bromide in the urinary bladder. The possibility of a purely local action of intravesically instilled emepronium bromide (Cetiprin) was investigated. Subjective reports and simultaneous urethrocystometry were used to assess such effect. In order to ascertain the purely local nature of effect, the serum was analyzed for freely circulating emepronium ion at increasing intravesical concentrations. No adverse effects were found at the highest dosage (100 mg in 100 ml saline solution), and the concentrations in serum were very low or were not detectable. The seemingly promising influence on micturition frequency and urge prompted a double-blind study of 20 patients with chronic urgency. In ten of them 100 mg emepronium bromide in 100 ml saline solution was instilled intravesically, while the other ten received only the same volume of saline solution. The symptoms were relieved in eight of the ten patients given emepronium bromide. The observation time was one week. The therapeutic possibilities were further evaluated in 24 women with frequency and urgency with or without urge incontinence. Emepronium bromide was intravesically instilled in the mentioned concentration and, if the symptoms were not totally relieved, this was repeated at weekly intervals. The maximum number of instillations was four and the observation time after the final treatment was at least three weeks. Frequency and urgency disappeared in approximately 70% of the affected women, nocturia in 60%, but urge incontinence in only 30%. Both frequency and urgency therefore were therapeutically influenced by intravesical instillation of emepronium bromide.


Renal allograft rupture.

Renal allograft rupture. In the present material of 448 consecutive renal transplants the incidence of allograft rupture was 3.6%. Among 389 first transplants there were two ruptures in the living donor group (126 patients, 1.5%) and 12 ruptures in the cadaveric donor group (263 patients, 4.6%). Only one ruptured kidney (living donor) achieved long term function. Four patients with first graft rupture were retransplanted with early loss of the kidney in all, in two because of rupture. Two of these patients received a 3rd cadaveric graft, of which one is functioning well after two years. All ruptures occurred within three weeks after transplantation, 14 kidneys ruptured during the first week. The clinical course and the operative findings suggested that rejection was the cause of rupture in all cases. This was confirmed by light and immunofluorescent microscopy of specimens from 15 kidneys, while one kidney only demonstrated extensive intrarenal vessel thrombosis. It is concluded that renal allograft rupture signals a strong immunological response in the recipient with poor graft prognosis. The chance of a successful retransplantation is small.


Renal graft rejection and blood transfusion before and during the transplant operation. The outcome of 390 consecutive primary cadaveric renal transplantations in a single centre was studied with regard to blood transfusions before and during the transplant operation. The graft loss due to rejection was 38% at 12 months in the transfused group compared to 67% in the non-transfused group (p less than 0.0005). When peroperative transfusions were added to patients in the previously transfused group, the graft loss was reduced to 34% compared to 50% if no peroperative blood was given (p less than 0.05).


Vascular function in kidneys preserved by simple cold storage in Sacks' or Collins' solutions. Vascular resistance was determined in 40 kidneys after preservation in Collins' or Sacks' solution for 24 and 48 hours. Kidneys without warm ischaemia and kidneys with up to 60 min of warm ischaemia prior to preservation were investigated, vascular resistance was determined during short time perfusion with Tis-U-Sol at 4 degrees. The experiments showed that cold preservation of kidneys with no warm ischaemia or 15 min of warm ischaemia gave a small increase in vascular resistance, independent of the solution used. 30 and 60 min of warm ischaemia gave a considerable increase in vascular resistance, but less increase was found in kidneys preserved in Sacks' solution. Determination of the weights and thereby formation of oedema showed slight dehydration in kidneys preserved in Sacks' solution. Kidneys preserved in Collins' solution showed small increase in weight. No correlation was, however, found between the vascular resistance and degree of oedema.


A new solution for initial perfusion of transplant kidneys. There is good evidence that blood should be removed from kidneys as completely as possible before they are transplanted. Rabbit kidneys, either freshly isolated or after one hour of warm ischaemia, were perfused with a solution of osmolality 400 mOsm/kg which contained high concentrations of Mg++ (72 mEq) and glucose (167 mM). The rate and completeness of blood removal were measured by collecting effluent fractions until 100 ml had been perfused, and then assaying the haemoglobin in each effluent fraction and the residual haemoglobin in the kidney. A pressure of 60 mmHg was found to be sufficient for effective perfusion. The addition of 5% dextran 40 (Rheomacrodex) to the perfusate greatly improved the completeness of blood removal from kidneys subjected to one hour of warm ischaemia. Rabbit kidneys washed out with this solution functioned excellently after transplantation.