Mechanisms of selective inhibition of 3' to 5' exonuclease activity of Escherichia coli DNA polymerase I by nucleoside 5'-monophosphates. The 3' to 5' exonuclease activity of Escherichia coli DNA polymerase I can be selectively inhibited by nucleoside 5'-monophosphates, wherease the DNA polymerase activity is not inhibited. The results of kinetic studies show that nucleotides containing a free 3'-hydroxy group and a 5'-phosphoryl group are competitive inhibitors of the 3' to 5' exonuclease. Previous studies by Huberman and Kornberg [Huberman, J., and Kornberg, A. (1970), J. Biol. Chem. 245, 5326] have demonstrated a binding site for nucleoside 5'-monophosphates on DNA polymerase I. The Kdissoc values for nucleoside 5'-monophosphates determined in that study are comparable to the Ki values determined in the present study, suggesting that the specific binding site for nucleoside 5'-monophosphates represents the inhibitor site of the 3' to 5' exonuclease activity. We propose that (1) the binding site for nucleoside 5'-monophosphates on DNA polymerase I may represent the product site of the 3' to 5' exonuclease activity. (2) the primer terminus site for the 3' to 5' exonuclease activity is distinct from the primer terminus site for the polymerase activity, and (3) nucleoside 5'-monophosphates bind at the primer terminus site for the 3' to 5' exonuclease activity.


Nonspecific interactions of Escherichia coli RNA polymerase with native and denatured DNA: differences in the binding behavior of core and holoenzyme. We have investigated the nonspecific interactions of Escherichia coli RNA polymerase core and holoenzyme with double-stranded (ds) and single-stranded (ss) DNA. Binding constants for these interactions as functions of such solution variables as monovalent and/or divalent cation concentration, temperature, or pH were determined by the method of deHaseth et a. [deHaseth, P.L., Gross, C.A., Burgess, R.R. and Record, M.T. (1977), Biochemistry 16, 4777--4783] from analysis of the elution of the proteins from small columns containing immobilized DNA. This technique, although as yet empirical, has been demonstrated to yield accurate binding constants fot the nonspecific interation of lac repressor with ds DNA. We find that observed binding constants (Kobsd) are extraordinarily sensitive functions of the monovalent cation concentration for the interactions of both core and holoenzyme with ds DNA. In the absence of divalent cations, the derivatives --(d log Kobsd/d log [Na+]) are 11 +/- 2 for the holo--ds DNA interaction and 21 +/- 3 for the core--ds DNA interaction. Consequently, approximately 11 and 21 low-molecular-weight ions are released, iin the thermodynamic sense, in the formation of the holo--ds and core--ds complexes, respectively (Record, M.T., Jr., Lohman, T.M., and deHaseth, P.L. (1976), J. Mol. Biol. 107, 145--158; Record, M.T., Jr., Anderson, C.F., and Lohman, T.M. (1978), Q. Rev. Biophys., in press). Ion release is a thermodynamic driving force for these nonspecific interactions and causes the stability of the complexes to increase very substantially with a reduction in monovalent ion concnetration. Possible molecular models which account for the different salt sensitivities of the holo--ds and core--ds complexes are discussed. Effects of the competitive ligand Mg2+ on these interactions are also examined. Substantial ion release (approximately 18 monovalent ions) also accompanies the interaction of either holo or core polymerase with ss DNA. Over the range of ion concentrations investigated the holo--ss interaction is substantially stronger than the core--ss interaction; furthermore, we conclude that the interactions of polymerase with ss DNA are, in general, stronger than the nonspecific interations of the enzyme with ds DNA. It is likely that the nonspecific interactions of RNA polymerase with DNA have physiological relevance. Not only is it plausible to assume that the same regions of the protein are involved in both specific and nonspecific interactions, but in addition nonspecific interactions of RNA polymerase and DNA may play role in determining the availability of this protein, in both the thermodynamic and the kinetic sense, for promoter binding and RNA chain initiation [von Hippel. P.H., Revzin, A., Gross, C.A., and Wang, A.C. (1974), Proc. Natl. Acad. Sci U.S.A. 71, 4808--4812]. Consequently, the strong dependences of the nonspecific interactions of RNA polymerase on ionic conditions suggest the possibility of a modulating role of ion concentrations in the control of transcription.


In vitro transscription of ribosomal RNA on phage lambdarifd 18 DNA. In vitro transcription of ribosomal RNA was studied on the DNA of the transducing bacteriphage lambdarifd 18, which carries an rRNA transcription unit from Escherichia coli. rRNA synthesis was preferential at all polymerase/DNA ratios tested, and at the optimal 0.3 weight ratio nearly 60% of the transcript was rRNA. At this ratio the principal product of transcription comigrated in acrylamide-agarose electrophoresis with authentic 30-S rRNA precursor synthesized in vivo. In the presence of rifampicin more than one equivalent of rRNA was synthesized, thus suggesting the existence of two initiation sites for rRNA on the phage DNA. Similar results were obtained on E. coli DNA. Preincubation with heparin virtually eliminated the transcription of rRNA, in sharp contrast with the results of similar experiments on bacterial DNA where rRNA genes were transcribed 4--5 times in the presence of heparin. The possible explanation of this difference between rRNA promotors on the phage and the bacterial DNA are discussed.


Unusual effects of 5a,6-anhydrotetracycline and other tetracyclines. Inhibition of guanosine 5'-diphosphate 3'-diphosphate metabolism, RNA accumulation and other growth-related processes in Escherichia coli. 5a,6-Anhydrotetracycline was discovered to be unique among several tetracycline derivatives tested in its ability to inhibit RNA accumulation in vivo at low concentration (20 microgram/ml and less). In addition, in vivo protein, DNA, and guanosine 5'-diphosphate 3'-diphosphate (ppGpp) synthesis were completely inhibited by 20 microgram/ml 5a,6-anhydrotetracycline. ppGpp decay in a spoT strain was inhibited by 20 microgram/ml 5a,6-anhydrotef RNA synthesis by a 5a,6-anhydrotetracycline may be due, in part, to reduced UTP and CTP synthesis. The effects of tetracyclines on in vitro ppGpp synthesis by crude stringent factor in the absence of ribosomes were investigated. It was determined that of six tetracyclines tested, four strongly inhibited the reaction (oxytetracycline, chlorotetracycline, dedimethylaminotetracycline, and tetracycline) whereas 5a,6-anhydrotetracycline gave a moderate inhibition and alpha-6-deoxyoxytetracycline resulted in only a slight reduction in ppGpp synthesis. It is proposed that tetracyclines interfere with factors involved in ppGpp metabolism and function.


Proteins associated with rRNA in the Escherichia coli ribosome. Ribosomal proteins located near the rRNA have been identified by cross linking to [14C]spermine with 1,5-difluoro-2,4-dinitrobenzene. The polyamine binds to double-stranded rRNA; those proteins showing radioactivity covalently bound after treatment with the bifunctional reagent should therefore be located in the vicinity of these regions of rRNA. Six proteins from the small subunit, S4, S5, S9, S18, S19 and S20 and ten proteins from the large subunit L2, L6, L13, L14, L16, L17, L18, L19, L22 and L27 preferentially take up the label. The results obtained with three proteins from the large subunit, L6, L16 and L27, show a high degree of variability that could reflect differences of conformation in the subunit population. Several proteins were drastically modified by the cross-linking agent but were not detected in the two-dimensional gel electrophoresis (e.g., S1, S11, S21, L7, L8 and L12) and therefore could not be studied.


Biological effects and repair of damage photoinduced by a derivative of psoralen substituted at the 3,4 reaction site: photoreactivity of this compound and lethal effect in yeast. A newly synthesized linear psoralen derivative, 3-carbethoxypsoralen is shown to bind to yeast nucleic acids after 365 nm light treatment. As compared to 8-methoxypsoralen, a well-known bifunctional furocoumarin, 3-carbethoxypsoralen exhibits a high photoaffinity for DNA in vivo. Both compounds bind and photoreact more efficiently in vivo than in vitro. In contrast to 8-methoxypsoralen, 3-carbethoxypsoralen does not form cross-links in yeast DNA as demonstrated by heat denaturation-reassociation studies at least in the range of doses used. Thus 3-carbethoxypsoralen reacts as a monofunctional compound. Wild-type cells of Saccharomyces cerevisiae are 6 times more resistant to 3-carbethoxypsoralen than to 8-methoxypsoralen plus 365 nm light treatment in terms of lethal effect. In comparison to angelicin, another monofunctional (but angular) furocoumarin, 3-carbethoxypsoralen is more photoreactive. When the photoaffinity for DNA of 8-methoxypsoralen and 3-carbethoxypsoralen are considered in relation to photoinduced cell killing, it is clear that monoadducts are very efficiently repaired in wild-type cells. In contrast to the additivity obtained with 8-methoxypsoralen, a synergistic interaction of the two different repair pathways blocked by the rad2 and the rad9 mutation is observed after 3-carbethoxypsoralen plus 365 nm light. Dark holding experiments show that the excision repair function which is present in wild-type and rad9-4 cells is important for dark recovery.


Pairing properties of the methylester of 5-carboxymethyl uridine in the wobble position of yeast tRNA3Arg. At optimum magnesium concentration (10 mM) both yeast tRNA1Arg and tRNA3Arg are able to bind to poly (A,G) and A-G-A in presence of Escherichia coli robisomes. With A-G-G only tRNA1Arg ginds, wherea tRNA3Arg (anticodon mcm5 U-C-U) is not bound. This result means that the methylcarboxymethyl substituant in position 5 of U prevents its wobble with G.


Strong magnesium binding sites in yeast phenylalanine transfer RNA. To ascertain the sites that are available for strong binding between magnesium ions and phosphate groups in yeast phenylalanine transfer RNA, all distances below 5.5 A separating the phosphoryl oxygens (Op) of the 76 nucleotide residues have been computed from the latest atomic coordinates for the monoclinic form of the tRNA crystallized in the presence of magnesium chloride. The 5.5 A distance is chosen as the upper limit expected for Op....Op distances involved in strong magnesium-phosphate binding, on the basis of studies on a model magnesium phosphodiester hydrate, taking into account the quoted standard deviation in the tRNA atomic coordinates. It is concluded that there are four possible sites for strong magnesium binding in the tRNA molecule, in addition to the three sites previously reported. One of the hypothetical sites: m2G10-OL, U47-OR, could be involved in the first stage of melting of the tRNA molecule, and may be relevant to tertiary structure stabilization, since it links the dihydrouridine arm with the extra (V) loop.


Yeast 3-phosphoglycerate kinase. Essential arginyl residues at the 3-phosphoglycerate binding site. Yeast 3-phosphoglycerate kinase (ATP:3-phospho-D-glycerate 1-phospho-transferase, EC 2.7.2.3) is inactivated by phenylglyoxal. Loss of activity correlates with the modification of two arginyl residues, both of which are protected by all of the substrates. The modification is not accompanied by any significant conformational change as determined by optical rotatory dispersion. Ultraviolet difference spectrophotometry indicates that the inactivated enzyme retains its capacity for binding the nucleotide substrates whereas the spectral perturbation characteristic of 3-phosphoglycerate binding is abolished in the modified enzyme. The data suggest that at least one of the two essential arginyl residues is located at or near the 3-phosphoglycerate binding site. A likely role of this residue could be its interaction with the negatively charged phosphate or carboxylate groups of 3-phosphoglycerate.


alpha-D-Mannosidase of Saccharomyces cerevisiae. Characterization and modulation of activity. A unique and interesting alpha-D-mannosidase (alpha-D-mannoside mannohydrolase EC 3.2.1.24) activity has been isolated from Saccharomyces cerevisiae. The enzyme was localized in a crude particulate fraction of the cell extract and was not solubilized by treatment with detergents or high ionic strength NaCl. The enzyme had a pH optimum of 6.3, Km 50 micron with p-nitrophenyl-alpha-D-mannopyranoside, and was competitively inhibited by D-mannose (Ki 20 mM). The enzyme is not affected by ethylenediaminetetraacetic acid, a number of different cations, or sulfhydryl reagents. It was inhibited by p-chloromercuriphenyl sulfonic acid and this inhibition is prevented by the addition of substrate. The cellular concentration of alpha-D-mannosidase is inversely proportional to growth rate, suggesting that the enzyme is under catabolite repression. The level of enzyme was found to increase approx. 8-fold during sporulation. This is apparently due to de novo synthesis, since inhibition of protein synthesis by cycloheximide prevents the increase in enzyme activity.


Affinity chromatography studies with the pyruvate dehydrogenase complex of wild-type Escherichia coli. 1. The lifetime of thiamine pyrophosphate-Sepharose 2B affinity matrices synthesized according to Matsuura et al. (Matsuura, A., Iwashina, A. and Nose, Y. (1973) Biochem. Biophys. Res. Commun. 51, 241-246) has been improved. The matrix interacts with bacterial pyruvate dehydrogenase complexes. 2. The synthesis of a stable thiochrome-Sepharose 2B matrix is described. 3. Both matrices bind the pyruvate dehydrogenase complex of Escherichia coli in a 50 mM phosphate buffer, pH 7.0. Elution is possibly by an increase in ionic strength but not by the cofactor or metal-cofactor complexes. 4. The presence of Mg2+, reduces the capacity of the affinity matrices but leads to higher specificity for the multienzyme complex. 5. The pyruvate dehydrogenase complex of E. coli has been successfully purified by combining a classical purification step with these affinity chromatography systems. The method is less suitable for large scale operation.


Purification and properties of the polymeric fatty acid synthetase from a filamentous fungus. Fatty acid synthetase was purified from the filamentous fungus, Aspergillus fumigatus to a specific activity of 4000--5000 munits/mg protein. Its purity was established by its appearance in electron micrographs, on sodium dodecyl sulphate polyacrylamide gels and by analytical ultracentrifugation, and also by its behaviour upon sucrose gradient centrifugation. This enzyme comprises two large polypeptides with molecular weights of 190 000 and 186 000. Evidence from electron microscopy indicates that it consists of three equivalent loops of protein. It dissociates into different-sized circular subunits on ageing or upon dissolution in buffer of low ionic strength. Differences in properties between this fungal synthetase and that found in yeast have been noted and relate, for example, to inhibition by acetyl CoA and malonyl-CoA, cold-lability and pH optimum. The synthetase from A. fumigatus, purified by different procedures, consistently exists in two forms of similar specific activity, with sedimentation coefficients approx. 40 S and 60 S. Synthetase activity present in crude extracts has been identified as a very heavy component with sedimentation coefficient greater than 100 S.


Function of phospholipids on the regulatory properties of solubilized and membrane-bound sn-glycerol-3-phosphate acyltransferase of Escherichia coli. Glycerophosphate acyltransferase (acyl-CoA:sn-glycerol-3-phosphate O-acyltransferase, EC 2.3.1.15) solubilized from Escherichia coli membranes was highly activated by phosphatidylglycerol. Phosphatidylethanolamine, cardiolipin and 1,2-diacyl-sn-glycerol 3-phosphate showed no effect. The Km of the enzyme for sn-glycerol 3-phosphate was increased 20-fold by solubilization. The value could not be restored by the addition of phospholipids. Temperature-sensitive regulation of the synthesis of either 1-palmitoyl- or cis-vaccenoyl-sn-glycerol 3-phosphate by the solubilized enzyme was identical with that by the membrane-bound enzyme in vivo and in vitro. The proportion of the molecular species of 1-acyl-sn-glycerol 3-phosphate varied when the ratios of palmitoyl-CoA and cis-vaccenoyl-CoA were changed, but changes in the sn-glycerol 3-phosphate concentration had no effect on selective acylation by both the solubilized and membrane-bound enzymes.


Folate polyglutamates in T4D bacteriophage and T4D-infected Escherichia coli. The folate compound which is a structural component of the Escherichia coli T-even bacteriophage baseplates, has been identified as the hexaglutamyl form of folic acid using a new chromatographic procedure (Baugh, C.M., Braverman, E. and Nair, M.G. (1974) Biochemistry 13, 4952-4957). It has also been found that the host cell contains a variety of polyglutamyl forms of folic acid. The major form is the triglutamate (about 50%) but small amounts of higher molecular weight folates including the octaglutamate (1.8%) have been identified. Upon infection with wild-type T4D bacteriophage there is a shift in the distribution of the folate compounds so that the folyl polyglutamyl compounds having the higher molecular weights are increased. Infection of E. coli with baseplate mutants of T4D containing an amber mutation in gene 28 resulted in the formation of significant amounts (over 7%) of folate compound(s) of molecular weight much higher than those observed either in uninfected cells or cells infected with wild-type T4D. It is suggested that the T4D gene 28 product functions to cleave glutamate residues from high molecular weight folyl polyglutamates to increase the availability of the folyl hexaglutamate for virus assembly.


[Glycoproteins and glycolipids in intercellular interactions]. Glycoproteins and glycolipids as important constituents of the extracellular matrix are considered in the present review. Their role in the processes of cell-to-cell contacts, control of cell proliferation and neoplactic transformation, are discussed. A glycosyltransferase-acceptor model of cellular recognition is also given.


[Ribosome binding to the membranes of Candida albicans]. The in vitro binding of free ribosomes to the membranes of polyenoic antibiotics-sensitive and resistant strains of Candida albicans, considerably differing in the composition of their sterol components, was studied. It was shown that the extent and type of membrane-ribosome interaction is similar in both cases. The dependence of binding on ribosome concentration in the incubation mixture is double-phase, as was shown for different membrane fractions of both sensitive and resistant strains. The role of lipid membrane components, particularly that of sterols in the membrane-ribosome interactions is discussed.


[Comparative study of the effect of ionic and non-ionic surface-active compounds on plasmic membrane of yeast protoplasts]. Lytic effect of some cationic surface-active compounds (SAC) on yeast protoplast plasmic membrane is studied. Comparative analysis of the lytic effect of anionic, cationic and non-ionic SAC is carried out, a correlation between key physico-chemical characteristics of the agents (critical concentration of micellization, hydrophyle-lipophyle balance) and their lytic activity is investigated. A proposition is made on the presence in all biphylic compounds of a hydrophyly gradient, which is due to the effect of hydrophylic group on lipophylic properties of hydrophobic part of the molecule, and possible role of this factor in thelytic effect of biphylic compounds on biological membranes is discussed.


[Animal tissue polynucleotide phosphorylase]. Localization, physico-chemical and catalytic properties and possible biological functions of polynucleotide phosphorylase (PNPase) from animal tissues are discussed. In animal tissue cells PNPase has multiple localization; the major amount of the enzyme is localized in the endoplasmic reticulum ribosomes. In the nuclei PNPase, similar to other endo- and exo-RNAses participates in the processing of precursor molecules of mature forms of RNA, whereas in the cytoplasm it is involved in the destruction of polyribosomes in the polyribosomes of rapidly growing tissues the activity of PNPase is extremely decreased. The mechanisms regulating the PNPase activity in rapidly growing tissues are discussed.


[Proteolysis of products of mitochondrial protein synthesis in isolated mitochondria of Saccharomyces cerevisiae yeasts]. Products of mitochondrial protein synthesis were specifically labeled with 3H-leucine in the presence of cycloheximide at the end of the exponential phase of yeast aerobic growth on glucose. The mitochondria isolated from these cells lost 37-40% of the label from the protein fraction during 60 min incubation at 35 degrees, which was accompanied by the accumulation of 3H-leucine in TCA-soluble fraction. This process was suppressed by phenyl-methyl sulfonyl fluoride and p-chloromercuriphenyl sulfonate, the inhibitors of proteases, and could thus be considered as the proteolysis of the products of mitochondrial protein synthesis. The proteolysis was ATP dependent and was stimulated by puromycine which is known to induce the removal of incomplete polypeptides from mitochondrial ribosomes. A body of indirect evidence allows a suggestion to be made that the observed proteolysis can hardly be due to the action of cytoplasmic proteinases.


[Spatial proximity of the ribosomal mRNA binding center to the 50S subunit]. 4-(N-2-chloroethyl-N-methylamino)benzylamide of 5'-heptaadenylic acid was used for affinity labelling of the ribosome in the vicinity of its mRNA-binding centre. This derivative, similar to the free oligonucleotide, stimulates the binding of [14C]-lysyl-tRNA to ribosomes of E. coli and alkylates ribosomes both the 30S and the 50S subunits. The alkylation of ribosomes is inhibited by pre-incubation of ribosomes with polyadenylic acid, which suggests that the chemical modification is a specific one and occurs in the vicinity of mRNA-binding site. The fact, that a short oligonucleotide having an active group on its 5'end attacks the 50S subunit of ribosome may indicate that the mRNA-binding centre is located in the contact region between ribosomal subunits.


[In vitro RNA-synthesis on immobilized DNA-templates]. The possibility of RNA-synthesis by E. coli RNA-polymerase using denatured DNA-templates from mouse liver, immobilized on nitrocellulose filters was shown. The size of RNA molecules, synthesized on immobilized templates was estimated by electrophoresis in polyacrylamide gel. The length of the RNA molecules was found to be about 30 nucleotides. Data from alkaline hydrolisys and sedimentation in sucrose density gradient suggest that there is no connection between the DNA-primer and the RNA-product, therefore the DNA-primer is not necessary for the initiation of RNA-synthesis.


[Isolation and physico-chemical properties of a complex kinetoplast DNA associate from the cells of Trypanosoma lewisi]. An associate of kinetoplast DNA (k-DNA) was isolated from the cells of Trypanosoma lewisi and characterized in terms of its sedimentation properties, melting parameters and reassociation kinetics. Electron microscopy studies showed that k-DNA isolated is a complex associate of circular molecules. The contour length of minicircular molecules is 0.77 mkm. k-DNA contains sites enriched by AT-pairs; melting of native associate and k-DNA fragments in the presence of 6.5 M sodium perchlorate results in the appearance of six zones within the temperature range of 47-64 degrees C. Data from k-DNA reassociation studies suggest that k-DNA associate constitutent molecules differ in sizes and nucleotide sequences. k-DNA is found to consist of two components with molecular weights of 1.7 . 10(6) and 17.5 . 10(6), respectively.


Burn wound coverings - a review.

Burn wound coverings - a review. Few injuries are more traumatic to the victim than a burn. To assist in preventing further damage beyond the burn site, a range of burn wound coverings has emerged over the years. These wound coverings, both natural and synthetic are discussed in connection with their historical significance and their effects on promoting the healing process. Particular reference is made to the choice of material for synthetic wound coverings.


[Role of lymphoid cells of the immune system in the genesis of kidney diseases]. Experiments on mice CBA X C57BL showed that the mixture of cells of the bone marrow, thymus, and spleen from donors with experimental hepatitis caused by carbon tetrachloride and from donors with resection of the normal liver, injected to healthy animals, caused dystrophic changes in the liver; this testifies to the presence of a special clone of cells with a damaging hepatotrophic effect, and participating in the mechanism of autoimmune disturbances in diseases of the liver.


[Several biological properties of E. coli SK cells]. Escherichia coli SK cells are not colicinogenic, but possess multiple resistance to antibiotics (tetracycline, kanamycin, penicillin, polymyxin, ampicillin). Numerous variations, sensitive to one or several of the antibiotics listed were obtained by cloning the initial culture. Experiments with acridine orange failed to eliminate completely the resistance of E. coli cells to all five of the antibiotics under study. Partial modifications in the spectrum of antibiotic resistance did not influence the host specificity system present in the cells SK. The capacity of restriction and the activity of the methylating enzymes in all the clones under study in the initial strain proved to be the same.


[Effect of stimulating antibody formation under the influence of bone marrow cells in vivo]. Injection of intact bone marrow cells to mice at the peak of the secondary immune response results in a 2.4-fold increase of the number of antibody-forming cells in the regional lymph node. Preliminary injection of bone marrow cells to donors of the immune lymph node cells decreases the stimulation effect of antibody formation when the lymph node cells are subsequently cultivated with the intact bone marrow cells. The data obtained demonstrate the cell interaction at the level of mature antibody producers in vivo.


[Correction of a T-immunodeficit in mice by bone marrow transplantation from donors treated with hydrocortisone]. Adult CBA mice were exposed to thymectomy, lethal irradiation, and protection by syngeneic bone marrow transplantation. In some experiments syngeneic bone marrow of donors, treated with hydrocortisone in a dose of 125 mg/kg for 3 days was used. The bone marrow of these donors contained cells with the Q-marker. Thymectomized and lethally irradiated animals subjected to the transplantation of syngeneic bone marrow from hydrocortisone-treated donors rejected the skin allotransplants, and the lymph node cells of these mice suppressed the endogenous colony-formation in the sublethally-irradiated hybrids (CBA X C57Bl/6) F1.


[Heterologous antisera to stromal mechanocytes of the bone marrow]. Heterologous rabbit antifibroblast serum (AFS) to stromal fibroblasts of the guinea-pig bone marrow was obtained. AFS was shown to bind specifically stromal fibroblasts and their precursors (but not macrophages) in the monolayer primary cultures of the bone marrow, thymus and spleen of guinea pigs (in the complement-dependent cytotoxic reaction and in the indirect immunofluorescence test); AFS also bound precursors of blood fibroblasts and peritoneal exudate (in the cytotoxic reaction). Precursors of the thymus fibroblasts were found to be more sensitive to the AFS action than the spleen and bone marrow precursors, this suggesting that the thymic stromal mechanocytes had a greater concentration of common tissue-specific antigens on their surface. Precursors of the stromal fibroblasts in native cell suspensions were found to be essentially more sensitive to the cytotoxic action of AFS than the colony-forming fibroblasts on the passage cultures.


[Autoaggressive immunocompetent cells in the bodies of mice at remote periods following irradiation]. Lethally irradiated DBA/l mice or (C57Bl X DBA/l1 F1 hybrid mice were injected with therapeutically effective doses of isologous bone marrow cells; simultaneously syngeneic lymph node cells from either intact (control) animals or mice survived after sublethal irradiation were transplanted. In control the viability of the recipients was not affected by the presence of lymphoid cells in the mixed transplant. In contrast, the beneficial action of the bone marrow cells was abolished (killing-effect) by the lymphoid cells from mice sacrificed 6 to 12 months after the irradiation (600--700 r). The manifestation of the killing-effect depended on the number of the transplanted lymphoid cells and on the dose of the bone marrow cells in the transplant. The killing-effect was not revealed when the lymphoid cells were obtained from the donors on the 30th day after irradiation. The results suggest the autosensitization of the organism at the late postirradiation periods.


[Suppression of the intradermal hypersensitivity reaction of the delayed type by a serum fraction from patients containing leukocyte migration inhibition factor]. The blood serum of patients with active tuberculosis of the lungs and chronic pneumonia inhibited migration of donor's leukocytes and macrophages of the peritonal exudate of guinea pigs when compared with migration of similar cells in the medium with the serum of cattle or donors. After chromatography these sera were fractionated on the columns with Sephadex G-100. Fractions containing the leukocyte migration inhibition factor (LMIF) suppressed, up to complete abolition, the intradermal reaction to tuberculin in man and guinea pigs sensitized with BCG. The LMIF is supposed to act in the regulation of delayed hypersensitivity reaction.


[Proliferation of the exocrine and endocrine portions of the pancreas after its resection]. A weak regenerative capacity of the pancreas was revealed after the resection of about 40 per cent of the tissue of this organ in mice-hybrids CBA X C57BL/6, weighing 20--28 g. During the 21 days of the experiment there was no significant restoration of the pancreas weight. An increase of the proliferative activity (the number of mitoses and daily fraction of the thymidine-3H-labeled cells were counted) was brief and failed to spread over the whole organ. The maximal number of the labeled nuclei in the epithelium of the acini and the islets was found in the area near the wound where the organ tissue was somewhat edematous. In the areas near the wound surface, but unaffected, the labeled cells count was increased during the early post-operative period only. The number of labeled cells of the islets and acini of the pancreas in the areas distant from the wound (in the loop of the duodenum) was no different from the control. The number of labeled cells in the islets was greater than in the acini.


[Stimulation of nonspecific resistance in mice with high- and low-molecular fractions of endotoxin on 2 models of Shigella infection]. The capacity of S. paratyphi B endotoxin and its high molecular (HMF) and low molecular (LMF) fractions to stimulate the resistance of mice to shigella infection in intraperitoneal and intranasal challenge was investigated. The HMF developed high activity after the intraperitoneal injection (efficiency index--EI = 2.8). All the preparations were less active on the lung model (EI = 1.6--1.7). Materials of this work lead to the conclusion that in studying the phenomenon of nonspecific resistance induction conditions providing antitoxic and antiinfectious resistance should be considered separately.


[Localization of labelled cells in the islets, acini and periinsular zone of the pancreas of mice undergoing resection]. Thymidine-3H was injected 5 times during 24 hours to mice CBA X C57BL/6 with resected pancreas (about 40 per cent of the organ tissue). The number of labeled cells in the islets and the acini failed to change with the prolongation of the interval after the last injection of thymidine-3H (5 to 18 days). Localization of labeled cells in the pancreatic islets at later periods showed not much difference from the same localization two hours after the isotope injection. Examination of large andmedium-sized islets at the maximal distance from the wound revealed a similar number of labeled cells per square unit in the central, middle andperipheral region of the islets 2 hours and 18 days after the last injection of the isotope. It is concluded that under conditions of this experiment no islet cells are formed due to external sources.


[Electron microscopic and autoradiographic study of the pancreas at different times of postmortem ischemia]. Experiments on dogs showed that the maximally admissible term of normothermic postmortem ischemia of the pancreas should not exceed 45 minutes, because irreversible destructive changes in the ultrastructural formations of the exocrine parenchyma cells developed later. The Langerhans islet cells proved to be more resistant to the ischemic effect.


[Morphology of the juxtaglomerular apparatus of allotransplanted human cadaver kidneys in the late periods following transplantation]. Histological structure of 13 allografted human cadaver kidneys was observed at late stages after transplantation (from 121 days to 3 years and 10 months). In the grafted kidneys with a well-preserved structure the microscopic picture of the juxtaglomerular apparatus (JGA) failed to differ significantly from the JGA in the control. This conclusion was confirmed by the karyometric data and by the results of the juxtaglomerular index calculation. In the allografted kidneys with severe destructive and dystrophic alterations there can occur a partial or complete JGA involution.


[Absnece of amyloid protein in the organs of normal mice]. The immunomorphological Coons' method (its indirect variant) was used to discover specific amyloid protein AA in the organs of mice with experimental amyloidosis and normal mice (adult, newborn, embryo). The use of pure antiprotein-AA-antibodies allowed to reveal the minimal deposits of amyloid protein at the earliest stages of amyloidogenesis. The organs of normal mice (adult, newborn, embryo) did not contain this protein.


Successful engraftment of cryopreserved autologous bone marrow in patients with malignant lymphoma. In an effort to evaluate the possible utility of cryopreserved autologous bone marrow infusions in man,  22 patients with malignant lymphoma resistant to conventional chemotherapy were treated with high-dose chemotherapy. This was followed in 12 patients by an infusion of their cryopreserved autologous bone marrow; 10 patients received chemotherapy alone and serve as controls. Following chemotherapy, severe leukopenia (less than 100 leukocytes/mm3) lasted 6-10 (median 8) days in patients receiving cryopreserved marrow, compared to 10-29 (median 16) days in controls (p less than 0.001). Recovery to 1000 leukocytes/mm3 occurred 10-18 (median 13) days after chemotherapy in autograft recipients but was delayed until 12-38 (median 23) days after chemotherapy in controls (p less than 0.001). Autografted patients also recovered granulocyte and platelet function significantly faster and had significantly fewer febrile days after chemotherapy than did controls. Cryopreserved autologous bone marrow infusions can hasten hemopoietic recovery in man after high-dose chemotherapy; this earlier reconstitution may be of clinical benefit to the patient.


Measurement of Donath-Landsteiner antibody-producing cells in idiopathic nonsyphilitic paroxysmal cold hemoglobinuria (PCH) in children. A 2-yr-old girl with a nonsyphilitic type of paroxysmal cold hemoglobinuria (PCH)  is reported. We applied the plaque-forming cell (PFC) assay to the measurement of Donath-Landsteiner (DL) antibody-producing cells in the patient. Some of the circulating B lymphocytes formed plaques in our assay system. Initially the number of PFC was 1442 +/- 225/10(6) lymphocytes and DL titer was 1:16. Thereafter the PFC decreased in number as the DL titer decreased. Eight weeks later circulating PFC were not found but the DL titer was still 1:1 positive. The PFC assay appears to be useful as a new method for obtaining rapid imformation as to the disease status of PCH.


Demonstration of the Fc-receptor of blood cells by soluble peroxidase-anti-peroxidase (PAP) complexes. The Fc-receptor of normal human leukocytes, of CLL-cells, and of hematopoietic cell lines was demonstrated with soluble peroxidase-anti-peroxidase (PAP) complexes. In about 9% of normal lymphocytes an almost continuous, strong labeling of the cell membrane was established. Some of these lymphocytes were characterized by a peculiar uniform fine structure. The percentage of PAP-labeled monocytes was in the range of 25%, neutrophils nearly 100%, eosinophils 0%, CLL-cells 10%. Labeled portions of the membrane were interiorized from monocytes. The lymphoid cell-line Daudi established from a Burkitt's lymphoma appeared almost negative, the cell line K562 established from a myeloid leukemia in 75% of the cells strongly positive. PAP-labeling was not influenced by preincubation with trypsine or with neuraminidase; it was negative when PAP-F(ab)2 was used. Results of PAP-labeling were not always in agreement with EA-rosettes or with agg-Ig.


An SDILINE profile oriented to patient care. Hospital physicians naturally require current medical information in order to treat their patients. An objectively obtained SDILINE storesearch was devised to obtain this information. A rank-ordered list of hospital discharge diagnoses, coded with ICDA terms, was translated into MeSH. The top two Bradford zones of journals defined as useful at the University of Missouri--Kansas City were ANDed to these MeSH terms. Citations retrieved from the storesearch are input into an in-house computerized data base. The method can be easily used by other medical or hospital libraries without access to computers.


A controlled trial of L-tryptophan in mania. A double-blind placebo-controlled trial of L-tryptophan, with pyridoxine and ascorbic acid, was carried out in 10 female patients suffering from mania. In contrast to earlier studies led by Prange and Murphy, L-tryptophan was found to be no better than placebo.


Haemolytic-uraemic syndrome complicating shigella dystentery in south Indian children. Shigella dysentery caused 65% of all cases of acute renal failure (ARF) seen in children treated at the Christian Medical College Hospital, Vellore, during the 33 months ending September 1977. In the 40 children with ARF secondary to shigella dysentery, haematological findings suggested that they were suffering from the haemolytic-uraemic syndrome, and glomerular hypercellularity and fibrin deposition were present in all 12 patients whose renal histology could be studied. Peritoneal dialysis was the main element of treatment: 43% of children who underwent dialysis improved, compared with only 25% of those who did not undergo dialysis. The haemolytic-uraemic syndrome precipitated by bacillary dystentery is therefore the most important cause of ARF in children aged under 5 years in Tamil Nadu and the adjoining area of Andhra Pradesh.


Sjörgren's syndrome treated with bromhexine: a randomised clinical study. Existing treatment for Sjögren's syndrome is unsatisfactory, and uncontrolled observations have suggested that bromhexine may be effective. Twenty-nine patients with Sjögren's syndrome were therefore assigned to two randomised double-blind crossover trials with bromhexine and placebo, each comprising two two-week periods. In the first trial bromhexine 24 mg/day was given by mouth; in the second the dose was increased to 48 mg/day. After each treatment period the Schirmer test response, break-up time, Bijsterveld score, and the time taken for the patient to eat a dry biscuit were recorded, as well as the patient's estimate of moistness in the eyes and mouth. In the second (higher-dose) trial values on the Schirmer test were significantly higher after bromhexine than after placebo and the break-up time was also increased after bromhexine, which suggested that the drug has a dose-dependent effect on lacrimal gland secretion in Sjögren's syndrome. It had no effect on salivary gland function. Bromhexine is therefore valuable in the treatment of Sjögren's syndrome.


Cell-mediated immunity during syphilis.

Cell-mediated immunity during syphilis. Evidence is presented which reinforces the complexity of the host-parasite interaction during the course of syphilis. Infection with Treponema pallidum evokes a complicated antibody response and an assortment of cell-mediated immune reactions in the host. It appears that humoral immunity plays a minor role towards the complete elimination of syphilitic infection while the cellular limb of the immune response may be an important host defence mechanism. Information now available indicates that a state of anergy, or immunosuppression, exists in the early stages of human and experimental rabbit syphilis based upon negative skin reactions to T. pallidum antigen(s), the abnormal histological appearance of lymphoid organs, and impaired in vitro lymphocyte reactivity. It is also evident that in the later stages of the disease cellular immunity becomes activated as delayed type skin reactions can normally be elicited in tertiary syphilitics and lymphocyte behaviour in cell culture appears normal. Several mechanisms have been invoked to explain the delay in an effective immune response against syphilitic infection and the duration of the disease: (1) a capsule-like substance on the outer surface of virulant T. pallidum may act as a barrier against treponemicidal antibody; (2) this material and other biological properties of virulent treponemes could enable spirochaetes to escape being engulfed by macrophages and other phagocytic cells; (3) antigenic competition among different treponemal antigens causing partial tolerance; (4) T. pallidum infection may bring about the elaboration of immunosuppressive substances of host or treponemal origin which inhibit the proper function of lymphocytes, macrophages, and other cell types.


TPHA test. Experience at the Clinic of Dermatology, University of Milan. The results of serological investigations with the treponemal haemagglutination (TPHA) test are reported in a large number of luetic and non-luetic patients. From these the following conclusions can be drawn: (1) The sensitivity of the TPHA test is decreased if the test is carried out with micromethods; however, in our opinion, the same degree of sensitivity can be obtained either with the macromethod at dilution 1/80-1/60 or with micromethods at 1/20-1/40. (2) The sensitivity of this test is high, being similar to that of the fluorescent treponemal antibody absorption (FTA-ABS) test in primary and secondary syphilis and even higher than that of the FTA-ABS test in treated subjects. (3) The specificity of the reaction is high, as demonstrated by examining sera in patients with a negative history and clinical examination together with negative results to Treponema pallidum immobilisation (TPI) and FTA-ABS tests, and by studying biological false positive sera. (4) For the serological screening, it may be sufficient to perform the TPHA test with the 1/20-1/40 micromethod together with the Venereal Disease Research Laboratory (VDRL) test. In patients with suspected syphilis, it is advisable to perform the TPHA test by the macromethod, in combination with the FTA-ABS test.


Dopamine uptake in serotoninergic terminals in vitro: a valuable tool for the histochemical differentiation of catecholaminergic and serotoninergic terminals in rat cerebral structures. An in vitro method was developed to separately visualize dopaminergic, noradrenergic and serotoninergic terminals in the cerebral, hippocampal and cerebellar cortices of the rat, by means of the glyoxylic histochemical fluorescence method. Animals were pretreated with alpha-methylparatyrosine to deplete catecholamine stores. Thin vibratome sections were made and incubated in the presence of various exogenous amines and inhibitors of the catecholaminergic and serotoninergic transport systems. Experimental conditions insuring the best specificity were determined and the validity of the combined pharmacological and histochemical approach was further tested by using animals in which the cortical dopaminergic, noradrenergic or serotoninergic innervations were destroyed. Under the experimental conditions used, norepinephrine as well as alpha-methyl-norepinephrine were taken up in only noradrenergic and dopaminergic terminals. A separate visualization of the two systems was obtained by using specific uptake inhibitors. Dopamine was taken up not only in catecholaminergic but also in serotominergic terminals. The uptake of DA in serotoninergic fibers was inhibited by a specific inhibitor of the serotoninergic transport system or by the presence of serotonine in the incubating medium. The signification and the implications of thes results are discussed.


Energy utilization in the induced release of gamma-aminobutyric acid from synaptosomes. Newly accumulated gamma-aminobutyric acid (GABA) was released from synaptosomes by treatment with 30 mM K+ or the Ca2+ ionophore A23187. Release was Ca2+-dependent and energy-dependent. The induced release of GABA was inhibited by S-13, an uncoupler of oxidative phosphorylation, by azide, a blocker of mitochondrial respiration, and by oligomycin, efrapeptin, tributyltin and dicyclohexylcarbodiimide (DCCD), which are inhibitors of Ca2+/Mg2+-ATPases, including mitochondrial ATPase. Efrapeptin blocked GABA release induced by K+ but not A23187-induced release. Azide and oligomycin appeared to inhibit GABA release as a consequence of their effects on mitochondrial ATP synthesis. However, the inhibition of GABA release by the other compounds could not be totally accounted for by their effects on synaptosomal ATP stores. It is proposed that these compounds, in addition to affecting ATP synthesis, directly affect biochemical reactions involved in GABA release. Thus, these and similar inhibitors seem to be useful probes of the transmitter release process.


Temporal changes in medial basal hypothalamic LH-RH correlated with plasma LH during the rat estrous cycle and following electrochemical stimulation of the medial preoptic area in pentobarbital-treated proestrous rats. In the present studies we have simultaneously measured changes in medial basal hypothalamic (MBH) leutenizing hormone-releasing hormone (LH-RH) and in plasma LH by radioimmunoassay in female rats at various hours during the 4-day estrous cycle and under experimental conditions known to alter pituitary LH secretion. In groups of rats decapitated at 12.00 h and 15.00 h on estrus and diestrus, plasma LH remained at basal levels (5-8 ng/ml) and MBH-LH-RH concentrations showed average steady state concentrations of 2231 +/- 205 pg/mg. On the day of proestrus hourly measurements of MBH-LH-RH between 12.00 h and 21.00 h suggested rhythmic rises and falls in the decapeptide concomitant with rises and falls in plasma LH. In a second group of pentobarbital-anesthetized proestrous rats a significant decline in MBH-LH-RH occurred (to 573 +/- 137 pg/mg) which then remained at low concentrations between 14.00 h and 18.00 h proestrus. Following bilateral preoptic area (MPOA) electrochemical stimulation of pentobarbital-treated proestrous rats, LH was significantly increased by 30 min, peaked between 90-120 min and returned to basal levels by 210 min poststimulation. In the same animals within 15 min poststimulation, MBH-LH-RH increased from the basal concentrations noted after pentobarbital anesthesia to elevated levels comparable to those observed throughout estrus, diestrus and on proestrous morning. Further, as plasma LH rose to peak concentrations and declined to basal plasma values, rhythmic rises and falls in MBH-LH-RH were observed with intervals between pulses of approximately 60 min. Seemingly, hypothalamic LH-RH is released as pulsatile pulses from a releasable pool; this pool is replenished and again LH-RH is discharged in response to constant stimulation by the preoptic brain.


The proportion of all point mutations which are unacceptable: an estimate based on hemoglobin amino acid and nucleotide sequences. Statistical analysis of the distribution of 156 kinds of human hemoglobin beta (Hbbeta) chain variants suggests that mutations are essentially random in their location. Thus differential fitness, not differential mutability, is the principal source of nonrandom distribution of interspecies differences in Hbbeta amino acid sequence. Similar analyses of both the location and the kind of interspecies differences detected among primates support this viewpoint and lead us to estimate that at least 95% of all amino acid subsitutions,i.e., nonsynonymous mutations, in Hbbeta are functionally unacceptable in homozygous state. Through the combined use of this estimate and the number of nonsynonymous and synonymous substitutions per nucleotide site inferred from comparisons of entire human and rabbit HbbetamRNA nucleotide sequences, we calculate (a) approximately 70% of synonymous Hbbeta mutations are adaptively undersirable and (b) the mutation rate underlying all changes is lesser than or equal to 10(-8) nucleotide substitutions per nucleotide site per year. Apart from such calculations, analyses of nucleotide patterns in HbbetamRNA as well as in rat preproinsulin mRNA reinforce the notion that a large portion of synonymous mutations are functionally unacceptable and rendered so by selective constraint, at a pretranslational level, of the abundance of particular nucleotide doublets such as CpG.


Biologic properties of bacterial lipopolysaccharides treated with chromium chloride. Addition of small amounts of chromium chloride to a saline suspension of Salmonella typhosa lipopolysaccharide (LPD; Difco) caused a marked reduction in several of the biologic activities of this substance including toxicity, B-cell mitogenicity, plasma colony-stimulating activity (CSA), radioprotective effect, and induction of the dermal Shwartzman reaction. Nevertheless, LPS treated with chromium chloride was found to be at least as effective as untreated LPS in enhancing resistance of B6CBF1 mice to the lethal effects of Klebsiella pneumoniae infection.


Enumeration of petroleum-degrading marine and estuarine microorganisms by the most probable number method. Several media designed for use in a most probable number (MPN) determination of petroleum-degrading microorganisms were compared. The best results, i.e., largest numbers, were obtained using a buffered (32 mM PO4=) liquid medium containing 1% hydrocarbon substrate. Of 104 presumptive oil degraders tested, 20 grew on oil agar medium but did not utilize oil or a mixture of pure paraffinic hydrocarbons (C10 to C16 n-alkanes) in liquid (MPN) medium. Visible turbidity in the liquid medium was correlated with hydrocarbon utilization. Counts of petroleum degraders obtained using liquid medium (MPN) were in most cases higher than those obtained on an oil-amended silica gel medium. Both procedures yield an estimation of oil degraders, and the oil-amended agar permits growth of organisms which do not degrade crude oil. All strains of oil-degrading microorganisms examined in this study were lipolytic, but the converse was not always true.


Inhibition of RNA polymerase from Bacillus thuringiensis and Escherichia coli by beta-exotoxin. The characteristics of exotoxin inhibition of deoxyribonucleic acid (DNA) dependent ribonucleic acid (RNA) polymerase isolated from Escherichia coli and Bacillus thuringiensis were investigated. RNA polymerase isolated from a variety of growth stages was partially purified and assayed using several different native and synthetic DNA templates, and exotoxin inhibition patterns were recorded for each. Although 8 to 20-h RNA polymerase extracts of E. coli retained normal sensitivity to exotoxin (50% inhibition at a concentration of 7.5 X 10(-6) M exotoxin), RNA polymerase isolated from late exponential and ensuing stationary-phase cultures of B. thuringiensis were nearly 50% less sensitive than exponential RNA polymerase activity. Inhibition patterns relating culture age at the time of RNA polymerase extraction to exotoxin inhibition suggested a direct correlation between diminishing exotoxin sensitivity and sporulation. Escherichia coli RNA polymerase could be made to mimic the B. thuringiensis exotoxin inhibition pattern by removal of sigma from the holoenzyme. After passage through phosphocellulose, exotoxin inhibition of the core polymerase was 30% less than the corresponding inhibition of E. coli holoenzyme. Heterologous enzyme reconstruction and assay were not possible due to loss of activity from the B. thuringiensis preparation during phosphocellulose chromatography, apparently from the removal of magnesium. In enzyme velocity studies, inhibition with exotoxin was noncompetitive with respect to the DNA template in the RNA polymerase reaction.


Incidence of the H2 group of plasmids in chloramphenicol-sensitive salmonella isolated in 1974 from clinical sources in Ontario. Chloramphenicol resistance in salmonella obtained from clinical sources in Ontario was previously found to be often mediated by R plasmids of the H2 incompatibility group. In the present study 40 salmonella strains resistant to one or more of kanamycin, streptomycin, sulfonamides, or tetracycline but sensitive to chloramphenicol, were investigated to determine if they contained R plasmids. Self-transmissible plasmids were isolated from 17 of the strains, and 7 of those showed the bacteriophage inhibition and thermosensitive mechanism of transfer characteristic of H2 plasmids. Entry exclusion and incompatibility experiments confiremd their classification. The results demonstrate that in this population of salmonella, R plasmids of the H2 group are prevalent. Experiments with plasmid-specific phages indicate that the plasmids of this sample, which are not in the H2 group, do not belong to any of the F, I, N, P, or W incompatibility groups.


Effect of sunlight on bacterial survival in transparent air samplers. Two types of commercially available microbiological air samplers (model STA-203, New Brunswick Scientific and All-Glass Impingers, Millipore) were used outdoors at an activated sludge treatment plant. The recovery of viable particles with both instruments was increased three- to eight-fold by shielding the trapping medium from the sun during the sampling process.


A rapid radiometric technique used to trap and quantitate 14CO2 evolved by slow-growing microorganisms. An apparatus to trap and quantitate 14CO2 is described. When used to measure antibiotic effects on Histoplasma capsulatum and Mycobacterium bovis (BCG), there was an inverse quantitative relationship between 14CO2 evolved and antibiotic concentration. The technique should prove useful for analyses that require trapping and quantitation of 14CO2, including antimicrobial sensitivities of slow-growing organisms.


Immunologic findings, thrombocytopenia and disease activity in lupus nephritis. Twenty patients with nephritis due to systemic lupus erythematosus were followed up for a mean of 34 months after renal biopsy with serial determinations of total serum complement and C3 and C4 concentrations, binding of deoxyribonucleic acid (DNA), antinuclear antibody pattern and platelet count. There were 25 episodes of nonhematologic observed disease activity in 16 of the 20 patients; elevated DNA binding and thrombocytopenia correlated well with these episodes. The mean platelet count during episodes of observed disease activity was 96 +/- 42 X 10(9)/L, which was significantly different from the mean count of 248 +/- 90 X 10(9)/L during disease quiescence. The proportion of false-positive results with the immunologic tests varied from 25% to 67% and with platelet counts it was 11%. It is suggested that thrombocytopenia may be a simple and accurate index of disease activity in lupus nephritis.


Tourette's syndrome: a treatable tic.

Tourette's syndrome: a treatable tic. Tourette's syndrome, or Gilles de la Tourette's disease, is a disorder characterized by involuntary tic-like muscular movements, compulsive behaviour and involuntary vocalization of sounds, words or profanities. It begins in childhood and may persist for life, with a varied pattern and course. Recent studies indicate an organic basis for the disorder, and an abnormality of dopamine or purine metabolism has been suggested. The treatment of choice is haloperidol administration; most patients do well with low or moderate doses for long periods. Because these patients are often mistakenly regarded as anxious, psychoneurotic or hysterical, correct diagnosis is important if they are to be treated appropriately and regarded properly in the home, school and society.


Pregnancy in patients with chronic renal disease. Pregnancy is not invariably contra-indicated in patients with pre-existing renal disease. Clinical data now exist that permit the clinician to distinguish such patients who are likely to experience difficulty during pregnancy from those in whom pregnancy can be undertaken with high expectation of success. Patients suffering from systemic lupus erythematosus, active or inactive, with or without lupus nephritis, should avoid pregnancy. Patients with other forms of chronic renal disease in whom the serum creatinine concentration prior to pregnancy is less than 1.5 mg/dL are not exposed to increased maternal or fetal risk. On the other hand, patients with serum creatinine values exceeding 1.6 mg/dL experience a high incidence of maternal and fetal complications and should avoid pregnancy. The life expectancy of recipients of a renal transplant is uncertain, and these patients should receive counselling as to the advisability of undertaking pregnancy. The maternal risk in such patients is not inordinately high, but the fetal risk is considerable.


Chronic granulocytic leukemia following successful renal transplantation. The use of immunosuppressive therapy has markedly increased over the past several years, and concomitant with its use has been an increased frequency of associated neoplasia. The patient presented is a 22-year-old white male who, following two renal transplants and prolonged immunosuppressive therapy with azathioprine and methylprednisolone, developed chronic granulocytic leukemia. Chromosome karyotyping demonstrated the somewhat unusual development of a Philadelphia chromosome with translocation to the No. 7 of the C group. A review of transplantation centers revealed that five cases of chronic granulocytic leukemia have occurred in a population of 25,000 renal transplant patients, a 5-fold increased incidence over the general population. Possible etiologies that may be responsible for the development of chronic granulocytic  leukemia in patients on immunosuppressive therapy are discussed. It is our hope that by the introduction of these reports of chronic granulocytic leukemia into the medical literature, the need for caution in the use of immunosuppressive drugs in nonmalignant disease will again be emphasized.


The epidemiology of breast cancer in 785 United States Caucasian women. A retrospective case-control hospital study of 785 Caucasian breast cancer patients and 2,231 age-stratified controls was conducted in New York City from 1969-1975. Patients were grouped by pre- peri- and postmenopausal status at diagnosis for the analysis to make a distinctive separation for variables showing a pre- and postmenopausal differential. Demographic characteristics were similar for cases and controls. Previously recorded hormone-related risk variables for this disease were largely confirmed for pre- and perimenopausal women, i.e., late age at first birth (greater than 25), premenstrual symptoms of breast swelling and premenopausal chills and flushes. Mother's history of breast cancer was also found to be a risk variable. Nulliparity was a risk factor only perimenopausally. No risk was foun for absolute height, weight or for obesity (Quetelet Index), prior breast diseases or previous usage of exogenous hormones of any type and no "protective" effect was found for multiparous women and for nursing. Perimenopausally diagnosed patients (menopause to 10 years after) were similar to premenopausally diagnosed women on most risk factors. Risk variables determined by this and other case-control studies cannot account for the magnitude of differences in the international incidence of breast cancer.


Metastatic malignant melanoma transplanted via a renal homograft: a case report. A case of transplantation of a malignant melanoma via a cadaver renal homograft is presented. The recipient developed a widespread malignancy ninteen months after renal transplantation. At autopsy the tumor was diagnosed as a malignant melanoma. Autopsy of the donor had previously revealed a spindle cell malignancy in the spleen. It is postulated that the splenic tumor in the donor represents a spindle cell variant of malignant melanoma, and hence this malignancy was inadvertently transferred to the recipient via the renal homograft.


Simultaneous extramedullary plasmacytomas of the vagina and vulva: a case report and review of the literature. An unusual case of simultaneous plasmacytomas of the vagina and vulva is presented along with a review of the literature on urogenital extramedullary plasmacytoma. The diagnosis in this case was obtained by vaginal cytological smear and later confirmed by histological examination of tumor biopsies. Of the four previously reported cases of malignant plasmacytoma of the vagina, one recurred locally and none was associated with metastases. In the case presented, local recurrence as well as multiple osseous metastases occurred.


T-lymphocytes and macrophages in primary murine fibrosarcomas at different stages in their progression. The relative contribution of lymphocytes, macrophages, and granulocytes to the cell content of primary 3-methyl-cholanthrene-induced murine fibrosarcomas was determined at different stages in their progression by differential cell analysis on enzyme-derived single-cell suspensions. Furthermore, immunohistological analyses were performed on the tumors to detect, quantitate, and determine the distribution of T-lymphocytes and macrophages. The T-lymphocyte content of small tumors was very high, and the T-cells were distributed throughout the tumor mass. As the tumor increased in size, there was a marked decrease in the relative T-cell content; most were located at the tumor periphery. Macrophages were present in significant numers in all tumors and appeared to increase in number as the tumors increased in size. Macrophages were distributed throughout the tumor mass, but generally they were more densely distributed near the tumor periphery. Granulocytes were present in low numbers in all tumors. Yeast phagocytosis was used to assess the functional capacity of the macrophage population. The phagocytic capacity of the macrophages was low in the small tumors, increased significantly as the tumors progressed, but dropped to relatively low levels in large tumors. The results represent a preliminary attempt to characterize the dynamics of host cell infiltration of primary immunogenic tumors.


Mechanisms of inhibition by ascorbate of microbial mutagenesis induced by N-nitroso compounds. Mutagenesis induced by N-methyl-N-nitrosoguanidine (MNNG) and dimethylnitrosamine (DMN) in Salmonella TA 1530 was inhibited by ascorbate. Inhibition of MNNG-induced mutagenesis resulted from a reaction between ascorbate and MNNG that led to consumption of MNNG. The rate of this reaction was considerably enhanced by catalytic amounts of Cu(II) and Fe(III). No direct reaction between DMN and ascorbate was detectable, but relatively high concentrations of Cu(II) enchanced inhibition of DMN-induced mutagenesis by ascorbate. Added protein reduced the effectiveness of Cu(II) as a catalyst of the reaction between ascorbate and MNNG, which suggested that the microsomal protein necessary to activate DMN, may reduce the concentration of free Cu(II) and thereby lower its catalytic efficiency. Mutagenesis by N-methyl-N-nitrosourea was not inhibited by ascorbate.


Mutagenicity of several classes of antitumor agents to Salmonella typhimurium TA98, TA100, and TA92. The mutagenic activities of antitumor agents, including 5 antibiotics, 19 antimetabolites, 5 alkylating agents, 2 alkaloids, 1 enzyme, and 1 adrenal steroid hormone, were tested on Salmonella tyhimurium TA100, TA98, and TA92. Four of these, busulfan, carbazilquinone, 1-(4-amino-2-methylpyrimidine-5-yl)methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride, and pipobroman were shown for the first time to be mutagenic. Further, the known mutagenicities of five others, daunomycin hydrochloride, Adriamycin hydrochloride, mitomycin C, 6-mercaptopurine, and cyclophosphamide, were confirmed.


Autologous bone marrow grafts in dogs treated with lethal doses of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea. Hematological and immunosuppressive effects of various single doses of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) were evaluated in the preclinical canine model. Immune function appeared not to be impaired by treatment with CCNU, but severe myelotoxicity contributed to the death of 3 of 4 dogs given 10 mg CCNU per kg and 5 of 5 dogs given 15 mg CCNU per kg within 10 days after drug administration. Infusions of autologous bone marrow protected 6 of 6 dogs receiving 15 mg CCNU per kg and 6 of 6 dogs receiving 20 mg CCNU per kg from lethal marrow failure. Dogs given 30 mg CCNU per kg and autologous marrow died within 7 days from severe gastrointestinal toxicity. We conclude that autologous bone marrow support may allow the use of high-dose CCNU regimens and thereby increase its therapeutic efficacy in the treatment of advanced cancer.


1,2-dihydro-1,2-dihydroxy-5-methylchrysene, a major activated metabolite of the environmental carcinogen 5-methylchrysene. The metabolic activation of the environmental carcinogen 5-methylchrysene was studied by combining high-pressure liquid chromatographic analysis of metabolites formed in vitro with assays of these metabolites for mutagenic activity toward Salmonella typhimurium. Metabolites were formed by incubation of 5-methylchrysene with the 9000 x g supernatant from Aroclor-treated rat livers. With the use of reverse-phase columns, the metabolites were resolved into nine peaks, A to I. Each peak was collected and tested for mutagenicity with activiation. Significant mutagenic activity was observed primarily in peak E and to a lesser extent in peak D. None of the other metabolites showed significant mutagenic activity. The major mutagenic metabolite (peak E) was identified as 1,2-dihydro-1,2-dihydroxy-5-methylchrysene (7.0% from 5-methylchrysene); Peak D was 7,8-dihydro-7,8-dihydroxy-5-methylchrysene (2.6% from 5-methylchrysene). Other metabolites included 9,10-dihydro-9,10-dihydroxy-5-methylchrysene, 9-hydroxy-5-methylchrysene, 7-hydroxy-5-methylchrysene, 1-hydroxy-5-methylchrysene, and 5-hydroxymethylchrysene. These results indicate that 1,2-dihydro-1,2-dihydroxy-5-methylchrysene is a major proximate mutagen of 5-methylchrysene.


Isophosphamide therapy for hematologic malignancies in patients refractory to prior treatment. Isophosphamide was administered to 27 patients with acute leukemia and to 15 patients with malignant lymphoma refractory to primary therapy. The starting dose of isophosphamide was 1200 mg/m2 administered as a daily continuous infusion x 5 days; the courses of treatment were repeated every 2-3 weeks. Of the 27 patients with acute leukemia, four achieved complete remission, two achieved partial remission, and two achieved hematologic improvement. However, no responses occurred in ten patients with acute myelogenous leukemia (AML). Thus, the response rate was 47% (eight responses among among 17 patients, in patients with acute lymphoblastic leukemia and acute undifferentiated leukemia. Seven of the 15 patients with malignant lymphoma responded. Most responses (five of six patients) occurred in patients with diffuse histiocytic lymphoma. Twenty-one of the 42 patients had received prior therapy with cyclophosphamide and 12 of these patients (two with leukemia and ten with lymphoma) responded, thus suggesting that as in the treatment of L1210 leukemia, isophosphamide is effective for tumors resistant to prior cyclophosphamide therapy. No significant genitourinary toxicity occurred; however, myelosuppression became the dose-limiting toxicity. Isophosphamide is active in malignant lymphomas and acute leukemias (except AML) and may have a role in combination regimens for such diseases.


A possible new approach to the treatment of metastatic breast cancer: massive doses of medroxyprogesterone acetate. The results obtained with a new hormone therapy using medroxyprogesterone acetate (MAP) in previously untested single and total doses in the treatment of advanced breast cancer are reported. Fifty-two postmenopausal patients were treated with an average total dose of 40 g of MAP for a period of 30 days. Nineteen of 44 patients (43%) had complete or partial remission, while the disease remained unchanged in nine of 44 patients (20%). Disease progression occurred in 12 of 44 patients (27%). Partial or complete remission occurred in 12 of 18 (67%) and four of six (67%) of the patients with dominant osseous and soft tissue metastases respectively. Three of ten (16%) of those with visceral metastases had remission. The average duration of remission was 7 months. Average survival times were 15.5 months for patients with remission, 8 months for those with no change, and 2.5 months for those with disease progression. From a subjective standpoint, pain was reduced significantly or disappeared in 34 of 36 patients (94%); this was also the case with respect to dyspnea (13 of 16 patients [81%]), anorexia (24 of 29 [83%]), asthenia (28 of 35 [80%]), and walking impairment (15 of 24 [63%]). When relapse occurred, patients previously treated with massive doses of MAP received further treatment with higher doses of MAP; four of 22 (18%) of the patients attained partial remission once again. Positive effects were also seen in subjective performance status, body weight, and EKG. We also describe the new clinical and toxicologic features of this treatment.


Combination chemotherapy plus methanol extracted residue of bacillus Calmette-Guérin or Corynebacterium parvum in stage III lung cancer. Seventy-six patients with stage III bronchogenic carcinoma were treated with monthly adriamycin, cyclophosphamide, cis-dichlorodiammineplatinum(II), and vincristine. In addition, they were randomized to receive either no immunotherapy, the methanol extracted residue of bacillus Calmette-Guérin (MER), or Corynebacterium parvum (C. parvum). Age, histology, extent of disease, prior therapy, and performance status were comparable in the three treatment groups. Twelve patients (16%) died having received only one treatment cycle. Sixty-four of the 76 patients received two or more treatments and were evaluable for response. There were no patients with complete responses, 14 patients (18%) with partial responses, 28 patients (37%) with no change, and 22 patients (29%) with progressive disease. Survival among patients receiving C. parvum is significantly longer (P less than 0.05) than that of patients receiving either MER or no immunotherapy.


Recurrent bladder papillomata treated with bacillus Calmette-Guérin: a preliminary report (phase I trial). Six patients with recurrent bladder papillomata were treated both by abdominal scarification with bacillus Calmette-Guérin (BCG) and by intravesical BCG, without resection. Four patients responded favorably (two showing major systemic complications), and the other two, who failed to respond, underwent further surgery.


Treatment of advanced ovarian cancer with cis=dichlorodiammineplatinum(II): poor-risk patients with intensive prior therapy. Nineteen patients with advanced ovarian adenocarcinoma were treated with cis-dichlorodiammineplatinum(II) (DDP), 50 mg/m2 as an iv bolus once every 3 weeks. Prior treatment had exhausted other chemotherapeutic options or damaged the bone marrow sufficiently to contra-indicate treatment with standard drugs. All patients had evaluable tumors and evidence of failure of prior therapy. DDP produced objective responses, relieved symptoms due to tumor, and improved the patients' quality of survival. There were no serious hematologic complications due to therapy except anemia.


Phase II trial of 5-fluorouracil, adriamycin, and mitomycin C in advanced colorectal cancer. Thirty-five patients with advanced measurable colorectal cancer were treated with a combination of 5-fluorouracil (5-FU), adriamycin, and mitomycin C (FAM). Objective regression of tumor was observed in six patients (17%); seven patients (20%) demonstrated stabilization of disease. The FAM regimen was generally well tolerated with toxic effects limited to bone marrow depression and mild-to-moderate nausea and vomiting. FAM combination chemotherapy did not significantly improve survival in responding patients, nor was it more effective than 5-FU alone in achieving objective regression of disease.


Chemo-immunotherapy for unresectable bronchogenic carcinoma. Seventy-nine patients with metastatic or unresectable bronchogenic carcinoma were treated with a regimen of combination chemotherapy which included methotrexate, Oncovin (vincristine), cyclophosphamide, and adriamycin (MOCA), and were randomized to receive no additional therapy, immunotherapy with bacillus Calmette-Guérin, or immunotherapy with Corynebacterium parvum (C. parvum). The response rate and estimated median survival time were 68% and 42 weeks in small cell carcinoma and 18% and 29 weeks in other histologic types. Improved survival correlated with high performance status and response to therapy. Immunotherapy did not improve response, time to progression, or hematopoietic tolerance of chemotherapy. C. parvum was associated with significant morbidity and was poorly tolerated. MOCA appears to be of modest value in the treatment of bronchogenic carcinoma, particularly of the small cell type. A role for immunotherapy remains unproven.


Phase II trial of ftorafur with mitomycin C versus ftorafur with methyl-CCNU in untreated colorectal cancer. In an attempt to substitute ftorafur for burdensome 5-fluorouracil (5-FU) infusions, 52 previously untreated patients were randomized to receive ftorafur with either mitomycin C or methyl-CCNU. Ftorafur was administered monthly as a 2-hour infusion daily x 5 days. Mitomycin C and methyl-CCNU were repeated every 8 weeks. A response rate of 27% (seven responses among 26 patients) was demonstrated on the mitomycin C arm compared to a response rate of 15% (four responses among 26 patients) on the methyl-CCNU arm (P = 0.25). There was no significant difference in the median survival between treatment arms. Central nervous system toxicity occurred in greater than 30% of the patients and appeared to be the limiting factor with ftorafur administration. Alternate schedules of ftorafur should be explored since there appears to be little advantage of a daily ftorafur schedule over conventional 5-FU infusions.


Altered pattern of vasopressin distribution in the hypothalamus of rats subjected to immobilization stress. An immunohistochemical study. Generally, the CRF-like activity of vasopressin is studied in experiments involving adrenalectomy and corticosteroid replacement. In order to avoid this complex type of stress, male and female (diestrus, estrus) rats were exposed to 5 min to immobilization stress and sacrificed 5, 15, and 30 min thereafter. After a survival period of 5 min the vasopressin-synthesizing part of the paraventricular nucleus exhibited an increased activity. Vasopressin-reactive axons in the pericapillary layer of the median eminence and among the solid cell clusters of the pars tuberalis became more conspicuous and increased in number. In this group of experimentally treated animals the prechiasmatic division of the supraoptic nucleus did not show any changes in immunoreactivity. The same holds true for the neurohypophyses in all experimental groups. In animals with increased survival times the supraoptic nucleus exhibited a slightly increased activity, whereas the staining intensity of the paraventricular nucleus decreases gradually. From these results it can be concluded that the paraventricular nucleus is involved in the first phase of the stress response. The problem of vasopressin or a very similar peptide synthesized in this nucleus and exerting a CRF-releasing function is discussed.


DNA sequence at the integration sites of the insertion element IS1. We have detected two independent occurrences of insertion mutations in the lacl gene of E. Coli, and have used small plasmids carrying the l gene to purify large amounts of DNA containing these insertions. Analyses with restriction endonucleases and DNA sequencing techniques establish that both insertions involve the previously characterized element IS1. In each case, the integration of IS1 into the l gene DNA is associated with a directly repeated sequence of 9 nucleotides appearing at each end of the insertion element. Since one of these sequences was present in the wild-type gene, the second sequence either preexisted in the IS1 before integration, or else was generated by the process of insertion itself. The 9 base repeat is different in both cases. We discuss the relevance of these findings to the mechanism of integration of transposable elements.


IS1 insertion generates duplication of a nine base pair sequence at its target site. Three independent integrations of the E. coli insertion sequence, IS1, into the gal operon have been analyzed. DNA sequences of portions of the wild-type galT gene which act as the target sites for these insertions, as well as the corresponding gal/IS1 junctions, are reported. Two features are particularly noteworthy. First, similar sequences appearing in inverted orientation consitute the ends of IS1: 18 of the terminal 23 base pairs at each end are identical. Second, in all three insertions, a 9 base pair segment found once in the wild-type sequence at the site of insertion is duplicated and appears in the same orientation at each end of the inserted element. The sequence of this 9 base pair repeat is different for each insertion analyzed. No homology between the inverted repeat sequences at the ends of IS1 and the sequences of the target sites is observed. Models for the mechanism of IS1 insertion are proposed.


Tubulin assembly protein: immunochemical and immunofluorescent studies on its function and distribution in microtubules and cultured cells. Cytoplasmic microtubule assembly from tubulin monomers requires an accessory protein or proteins present is isolated microtubules. These proteins have been designated "tau" factors. One such factor, tubulin assembly protein (TAP), has been purified to homogeneity from calf brain microtubules. A precipitating, monospecific antibody against the protein has been prepared. The antibody has been used to investigate the mechanism of TAP action in microtubule assembly and the distribution of TAP in cellular microtubules. Immunochemical, immunofluorescent and electron microscopic studies indicate that TAP functions stoichiometrically by binding physically to tubulin to form a complex active in microtubule assembly. TAP is an elongation protein which is required throughout the growth of a microtubule and which is actually present along the entire microtubule. Immunofluorescence microscopy has been used to demonstrate that TAP is distributed throughout the cytoplasmic microtubule network of cultured human, hamster and rat cells-both normal and virally transformed. Immunofluorescence of cells in mitosis shows that TAP is present in the mitotic spindle. These results demonstrate the biological importance of tubulin assembly protein and suggest that it or immunologically related "tau" proteins represent ubiquitous cofactors in cytoplasmic microtubule assembly.


Tubulin and actin in paired nonneoplastic and spontaneously transformed neoplastic cell lines in vitro: fluorescent antibody studies. Pairs of nonneoplastic and spontaneously transformed neoplastic cells were derived from rat, mouse and hamster embryos. The neoplastic cells of each pair had poorly spread cellular morphology, grew in agarose in vitro and produced invasive sarcomas in vivo; the nonneoplastic cells exhibited none of these properties. The distribution of microtubules and microfilament bundles (stress fibers or actin cables) was examined in five such paired lines and in 3T3 and SV40-transformed 3T3 cells by indirect immunofluorescent microscopy of fixed cells treated with rabbit antibody prepared against bovine brain tubulin or guinea pig smooth muscle actin, respectively. Actin cables in all the neoplastic cells appeared thinner and more sparse than in the paired nonneoplastic cells. These differences were also observed in living cells with polarization microscopy. In contrast, microtubules appeared similar in neoplastic and nonneoplastic cells, both in areas of thin peripheral lamellar cytoplasm which allowed a clear visualization of fine, curving microtubules and in regions of thick, central endoplasm which obsecured individual microtubules. In fact, the main morphological difference between neoplastic and nonneoplastic cells was the relative amount of lamellar cytoplasm or endoplasm, rather than the appearance of microtubles in either region. Thus the distinctive growth properties and retracted cellular morphology of neoplastic cells in this study did not correlate with decreased or disorganized microtubules, but with thin and sparse actin cables.


The effect of proteases on plasmodium formation in Didymium iridis. An improved assay for quantitatively measuring the number of plasmodia formed with time is presented. Using this assay we have investigated the effects of three proteases, subtilisin PBN', subtilisin carlsberg and alpha-chymotrypsin. We have shown that 1) plasmodium formation is sensitive to protease treatment only during the first 2 h after mixing amoebae of compatible mating type but not after, 2) amoebae are protease sensitive when treated 1 h prior to mixing, 3) the two clones used have different sensitivities to protease treatment and 4) these effects are due to enzymatic activity and have little effect on viability. The meaning of these results in relation to recent evidence for a diffusible inducer of plasmodium formation is discussed.


Sporulation competence in Aspergillus nidulans: a role for iron in development. There is a difference in the response of DNA from mycelial extracts of Aspergillus nidulans to hot acid hydrolysis depending upon the state of sporulation competence. The DNA in incompetent culture mycelia is not hydrolyzable while the DNA in competent culture is hydrolyzable. The inhibition of DNA hydrolysis is due to the presence of iron. Although the concentration of iron decreases in mycelia during growth, there is sufficient iron present in competent mycelia to inhibit DNA hydrolysis. The change in DNA hydrolyzability may be the result of a change in intracellular iron distribution, or a change in an iron binding component. We suggest that these changes are related to the altered capacity for gene expression which occurs at the time of acquisition of sporulation competence.


[Treatment of renal carcinoma with metastases]. They stress the need to evaluate certain characteristic of the tumour metastasis in the light of a series of limit events pin-pointing the lack of understanding of the natural history of renal carcinoma. They stress the need to evaluate certain characteristic of the tumour which are not normally taken into consideration by the surgeon: rate of proliferation, monitoring of hormonal receptors, and immunological monitoring. They reach the conclusion that treatment of renal carcinoma with metastasis must be the outcome of multidisciplinary planning.


Comparison of the EMIT test with high-performance liquid chromatography for the determination of phenobarbital in serum. The new enzyme immunoassay technique (EMIT) is compared to high-performance liquid chromatography used for determining phenobarbital in serum. After resuming the methods of both techniques, we compare the results of 116 sera collected from patients receiving treatment for epilepsy. The equation of the correlation curve is: HPLC = 0.98 EMIT + 0.12. The reliability characteristics are comparable. The enzyme immunoassay technique is less specific but more practicable.


An assessment of the radial diffusion method for the measurement of pepsin in gastric secretion and its comparison with the haemoglobin substrate colorimetric method. The radial diffusion method of pepsin assay has been compared with a modified Anson-Mirsky colorimetric method. Though claimed to measure accurately over a wide range, we found that the assay range of the radial diffusion method was only 10 to 100 mg/l; even with these narrow limits, it was inaccurate. In contrast, the colorimetric method was much more rapid, sensitive and could accurately measure a wide range of 0.5 to 500 mg/l. The colorimetric technique is therefore the superior method.


Immunoglobulin systems of human tonsils. II. Patients with chronic tonsillitis or tonsillar hyperplasia: quantification of Ig-producing cells, tonsillar morphometry and serum Ig concentrations. Patients suffering from recurrent tonsillitis (RT), hyperplastic tonsillitis (HT) or idiopathic tonsillar hyperplasia (ITH) were compared in immunological studies with subjects showing clinically normal palatine tonsils. Serum concentrations of immunoglobulins, particularly IgG, were increased in association with tonsillitis. Conversely, the number of IgG-, IgA- and IgM-producing immunocytes was reduced per tonsillar tissue unit. The density of such cells was decreased in the germinal centres of the lymphoid follicles, in the extrafollicular areas and in the reticular parts of the crypt epithelium. However, only small changes in immunocyte class ratios were observed, excepting a significantly raised proportion of IgD-producing cells. The immunohistochemical results indicated a defect in the tonsillar immunological function associated with disease, perhaps as a result of inadequate  stimulatory signals for local maturation of B-cell clones. An influence of low age on this result in the HT and ITH groups could not be excluded. In the RT group there was a significant negative correlation between the tonsillar density of IgA-producing cells and the rate of synthesis of serum IgA. This may indicate that compensatory mechanisms are reflected in inversely related levels of local and systemic IgA synthesis.


Comparison of a leucocyte adherence test with the leucocyte migration inhibition test and skin reactivity to PPD. Using PPD as antigen, a leucocyte adherence test was compared with the leucocyte migration inhibition test and with the results of skin testing in healthy subjects. In the presence of antigen, leucocyte adherence was significantly greater in skin test-positive subjects than in skin test-negative subjects who had not previously had BCG vaccination. When the test was performed in heterologous serum, skin test-negative subjects who had had previous BCG vaccination showed significantly greater leucocyte adherence in the presence of antigen than skin test-negative subjects who had not had BCG vaccination; this difference was not present in autologous serum and its abolition was attributed to a serum blocking effect. The leucocyte adherence test correlated with the results of the leucocyte migration inhibition test in heterologous serum, and also with the results of skin testing interpreted in the light of previous vaccination history. The finding of increased rather than decreased adherence in the presence of antigen, although previously reported, is unusual.


Differing surface marker characteristics in plasma cell dyscrasias with particular reference to IgM myeloma. Immunological marker studies using direct immunofluorescence and rosette methods were performed on the bone marrow and peripheral blood of an IgM myeloma, and on the peripheral blood of two cases of IgA myeloma and one IgG myeloma. These studies confirmed the peripheral blood involvement in plasma cell dyscrasias. In the IgA myeloma, the membrane and cytoplasmic immunoglobulin was restricted to IgA, and the IgG myeloma did not express immunoglobulin at the surface of the cells shown to contain cytoplasmic IgG. The IgM myeloma cells, on the other hand, expressed membrane immunoglobulin, including IgD, of a single light chain type. The majority of plasma cells secreting IgA or IgG did not express the IgG Fc receptor, but most of the cells from the IgM myeloma, including plasma cells, did express IgG Fc receptors.


B- and T-lymphocytes in toxic diffuse goitre. Operation specimens of thyroid tissue from patients with Graves' disease were digested, with collagenase, down to a cell suspension. The lymphocytes in the suspension were concentrated by density-gradient sedimentation and the proportions of B and T lymphocytes were determined. Six patients with Graves' disease were studied. The proportions of B and T lymphocytes in the thyroid glands were similar to the proportions of B and T lymphocytes in the venous blood from healthy subjects.


Adverse effect of splenectomy in renal transplantation. Between January 1968 and June 1974 at Newcastle upon Tyne, 63 patients underwent splenectomy in association with transplantation; 45 of these had splenectomy with bilateral nephrectomy before (20) or at the time of (25) transplantation; 18 had post-transplant splenectomy for leucopenia. Mortality was significantly higher in splenectomized patients than in 136 non-splenectomized controls. Of the 63 splenectomized patients, 25 died within 1 year of transplantation, 12 of severe infection associated with leucopenia. Although splenectomy produced a temporary rise in white cell count, leucopenia during the first year of immunosuppressive therapy was not significantly less frequent in splenectomized patients than in controls. There was no significant difference in graft loss between the splenectomy and control groups. It is concluded that splenectomy is contra-indicated in patients who are to undergo renal transplantation and confers no benefit when carried out because of leucopenia developing after renal transplantation.


De novo development of membranous nephropathy in cadaver renal allografts. Glomerulonephritis in transplant recipients often reflects recurrence of the immunopathogenetic mechanism causing the original renal disease. Membranous nephropathy (MN), a progressive immune complex mediated glomerular disease and the commonest cause of idiopathic nephrotic syndrome in adults, has been virtually unreported in transplant recipients. Two cases are reported here of typical MN (by clinical, light, immunofluorescent and electron microscopic criteria) developing de novo in the transplants of patients whose original diseases were anti-GBM nephritis and focal glomerular sclerosis. NM developed following episodes of viral hepatitis and renal infarction respectively. Possible mechanisms by which this lesion might develop in these patients are investigated and discussed. Chronic immune complex nephropathy (MN) can develop de novo in immunosuppressed transplant recipients apparently initiated by events in the post-transplant period.


Neonatal coagulation: normal physiology and pathophysiology. The newborn infant, particularly when premature, has a haemostatic mechanism which may not be entirely capable of withstanding the onslaughts of trauma, infection, asphyxia or other complications of the neonatal period. He is at risk of local or diffuse haemorrhage, which may at times be serious or even life-threatening. The cause of haemorrhage during the newborn period can generally be ascertained by a careful history and brief physical examination directed toward recognition of any predisposing factors or underlying diseases. Screening laboratory tests can usually be correctly interpreted as long as certain laboratory artifacts and physiological peculiarities of the neonatal coagulation mechanism are kept in mind. Diagnosis of and therapy for vitamin K deficiency and haemophilia in the healthy-appearing neonate is generally carried out with little difficulty. The seriously ill neonate with bacterial sepsis, respiratory distress syndrome, or extreme immaturity presents greater problems, for laboratory tests may be more difficult to obtain and interpret and underlying conditions may be untreatable. DIC occurs commonly in such neonates, and transfusion therapy, with or without heparin, is often unsuccessful. A persistent dilemma are those neonates with fatal intravascular haemorrhage, in whom definable haemostatic abnormalities are few and transfusion therapy is futile.


Gallium-67-citrate accumulation in the heart with tumor involvement. Tumor involvement of the heart is often found in post-mortem examination of patients who died of widespread malignancy. Recently, we saw four patients who had intense gallium uptake in the region of the heart. All of them had some cardiac symptoms and abnormal physical findings. Tumor was found in the pericardium and myocardium in all three patients who received necropsy.


Aspirin and aspirin-caffeine in postpartum pain relief. In two double-blind randomized, balanced, single oral dose studies with 140 women, a combination of 800 mg aspirin and 64 mg caffeine (aspirin-caffeine) was compared to 650 mg aspirin and to placebo. In patients with moderate to severe uterine or episiotomy pain, there was greater analgesic response with active drugs when the initial pain intensity was more severe. In patients with severe episiotomy pain, aspirin-caffeine was more effective than 650 mg aspirin (p less than 0.05) at the second and third hours. There was no difference between analgesic effects of aspirin and aspirin-caffeine in women with severe uterine pain.


Clinical pharmacokinetics of lignocaine. Lignocaine is widely used as a local anaesthetic and antiarrhythmic drug. It is commonly administered to patients with acute myocardial infarction as prophylaxis for ventricular fibrillation, although its efficacy in preventing primary ventricular fibrillation is still debated. Toxicity, sometimes with serious clinical consequence, is not uncommom and is usually related to overdosage. Blood lignocaine concentrations correlate roughly with antiarrhythmic and toxic effects and might be useful as an end point for monitoring prophylactic therapy. Administration of lignocaine as a local anaesthetic may result in blood lignocaine concentration in the antiarrhythmic or even toxic ranges. Expected peak levels for various routes of local anaesthesia are tabulated so that 'safe' total doses can be calculated. Intramuscular injection of high doses results in sustained therapeutic levels but is often associated with early minor toxicity. Lignocaine is eliminated primarily by hepatic metabolism, which appears to be limited by liver perfusion. Active metabolites may contribute to therapeutic and/or toxic effects. Disease states such as cardiac failure or drugs that alter hepatic blood flow may significantly affect lignocaine clearance. Pharmacokinetic studies in man show wide variability in drug disposition between patients, even when cardiac and hepatic status is considered, making specific dosing recommendations a problem. With intravenous injection, multicompartment kinetics is observed, with an initial rapid decline phase and initial decline in antiarrhythmic activity due to redistribution. With constant infusion, steady state concentrations of lignocaine are seen after 3 to 4 hours in normal subjects and after 8 to 10 hours in patients with myocardial infarction without circulatory insufficiency. In patients with cardiac failure, blood lignocaine concentration may continue to rise for 24 to 48 hours. In the presence of cardiac failure, decreased volumes of distribution and clearance require reduction in loading and maintenance doses. Lignocaine clearance is reduced in patients with liver disease and appears to be a sensitive index of liver dysfunction. A dosing algorithm for treatment of patients with myocardial infarction is presented.


National Clearinghouse for Poison Control Centers. The Clearinghouse was originally conceived to coordinate information flow among the then few Poison Control Centers. Services were to be provided through the state health departments where complete control over the individual state's poison centers was to reside. No authority to establish or regulate centers, or to set standards for staffing and operation of centers, or to designate regional centers was granted. However, growth in the numbers of centers, more sophistocation in the state of the art, more demands for product and human experience data, calls for research support, regionalization, standards setting, competency testing, and others has led to an increasing call for more and more Clearinghouse involvement in the entire poison control program. Though staffing and funding remains fairly constant, the Clearinghouse hopes to be as involved and to provide as great a service as possible to those involved in clinical toxicology and poison control.