Rheumatol Int, 1986, 6(3), 139 - 44 Measurement of serum DNA binding in chronic active hepatitis and systemic lupus erythematosus using the Farr assay; Pollard KM et al.; Antibodies to double-stranded DNA (dsDNA), generally regarded as highly specific for systemic lupus erythematosus (SLE), have also been reported in chronic active hepatitis (CAH) . Using the Farr assay and E . coli DNA, fractionated by benzoylated-naphthoylated-DEAE cellulose chromatography into dsDNA and dsDNA containing single-stranded regions, we compared the serum DNA binding of CAH and SLE patients . Although CAH sera were found to have dsDNA binding significantly above the normal control group such binding was of low level and we could find no evidence of markedly elevated dsDNA binding in CAH . However 12 of the 20 CAH sera studied did bind preferentially to dsDNA containing single-stranded regions suggesting the presence of anti-single-stranded DNA antibodies . We conclude that the description of elevated anti-dsDNA antibodies, as measured by the Farr assay, in CAH is due to the interaction of anti-single-stranded DNA antibodies or other serum components with single-stranded DNA contaminating dsDNA preparations. Gene, 1986, 43(3), 311 - 7 Synthesis of fused glycoprotein D of herpes simplex virus type 1 but not type 2 inhibits Escherichia coli hosts; Steinberg DA et al.; Glycoprotein D from either Herpes simplex virus type 1 (gD-1) or type 2 (gD-2) has been expressed in Escherichia coli as a series of chimeric proteins . The expression vector used in this study, pJS413, was derived from pBR322 and contains several cloning sites between the lacZ promoter-operator and the phage lambda cro gene . Plasmids containing fusions between the cro gene, gD-related sequences and lacZ was constructed and shown to direct the synthesis of 160-kDa proteins . The accumulation of fusion protein could be visualized as inclusion bodies when the cells were examined by dark phase-contrast or transmission electron microscopy . None of the plasmids that encoded cro::gD gene fusions yielded significant amounts of material upon induction with isopropyl-beta-D-thiogalactopyranoside . In addition, certain plasmids produced a form of Cro-gD-1 fusion protein which resulted in severe growth inhibition of E . coli . These inhibitory effects were attributed to the presence of specific gD-1 sequences, i.e., the transmembrane and cytoplasmic anchor region of the protein. Gene, 1986, 43(3), 287 - 93 Nucleotide sequence and deduced amino acid sequence of Escherichia coli adenine phosphoribosyltransferase and comparison with other analogous enzymes; Hershey HV et al.; The Escherichia coli apt gene has been analyzed and its nucleotide (nt) sequence and the deduced amino acid (aa) sequence compared to those of other phosphoribosyltransferases (PRTs) . The apt mRNA has a 102-nt leader sequence which may form alternate secondary structures . The RNA transcript may also form several 3' hairpin structures, which, however, do not appear to act as Rho-independent terminators . All PRTs, including E . coli adenine PRT (APRT), have a strongly conserved 13-aa sequence, as well as other regions of aa sequence or structural similarity . E . coli APRT is remarkably similar to the mouse enzyme. Gene, 1986, 43(3), 183 - 96 Overlapping arrangement of the recF and dnaN operons of Escherichia coli; positive and negative control sequences; Armengod ME et al.; The recF gene of Escherichia coli controls one of the recombination pathways and UV sensitivity, but its precise function and expression pattern are still largely unknown . We have characterized the promoter region of the recF gene by mapping for E . coli RNA polymerase binding sites, in vitro transcription experiments, cloning, and S1 mapping of in vivo mRNAs . It contains three overlapping promoters, two initiating transcription towards recF and one in the opposite direction . The recF promoter region is located about 600 bp upstream from the start codon of the recF structural gene and resides entirely within the translated region of the preceding gene, dnaN, which encodes for the beta subunit of DNA polymerase III . This unusual arrangement might provide discoordinate regulation of the recF and dnaN genes, thus controlling the level of DNA polymerase III holoenzyme . Expression of recF is also negatively controlled by sequences located upstream as well as inside the recF coding frame . Such negative regulation may serve to prevent toxic effects due to accumulation of an excessive number of copies of the recF gene product. Circ Shock, 1986, 19(4), 409 - 22 The pulmonary microvascular response to infusion of live Escherichia coli in sheep with acutely or chronically prepared lung lymph fistula; Smith L et al.; The effects of infusion of live Escherichia coli bacteria in awake sheep with a chronic lung lymph fistula (n = 15) were compared to anesthetized animals (n = 7) receiving the same septic insult after surgical trauma including bilateral thoracotomies for lung lymph cannulation (acute group) . During preseptic baseline conditions, pulmonary arterial pressure (Ppa) and central venous pressure (Pcv) were increased and leukocytes decreased in the newly operated animals compared to the sheep with a chronic lung lymph fistula . After i.v . infusion of live E . coli 10(9) X kg-1 b.w . over 20 min, arterial pressure (Psa), cardiac output (Qt), leukocytes, and partial pressure of arterial oxygen (PaO2) decreased in both groups . Ppa peaked after 15 min at 37.2 +/- 2.5 in the chronic and 33.4 +/- 3.3 mm Hg in the acute group . In the chronic group, Ppa remained elevated but not in the acute group during the rest of experiment . Lung lymph flow (QL) increased significantly in both groups during the initial high Ppa, but it increased to a higher level in the chronic group . After 150 min, QL did not differ between the groups but remained elevated over baseline . Lymph-to-plasma concentration ratio (L/P) for total protein decreased in the chronic group during the initial high QL . This decrease was not seen in the acute group that had a significantly higher L/P between 30 and 120 min after sepsis . The high QL with unchanged L/P compared to baseline indicated increased permeability in the pulmonary microvessels in both groups but the changes in permeability, hemodynamics, or respiratory parameters after sepsis were not aggravated by the surgical trauma. Basic Life Sci, 1986, 38, 439 - 52 Use of gradient denaturing gels to determine mutational spectrum in human cells; Cariello NF et al.; Based on the fact that mutagens induce specific patterns of gene mutations, this paper outlines a method to allow discrimination among mutagen-treated populations . The technique should allow direct screening of human tissue for genetic change, using human peripheral blood lymphocytes deficient in the enzyme hypoxanthine guanine phosphoribosyl transferase . The method is based on gradient denaturing gel electrophoresis, which separates short DNA molecules according to their melting properties . The melting behavior of DNA fragments is extremely sequence-dependent, and DNAs with single basepair substitutions often migrate differently . Even DNA fragments with the same basepair substitutions at different locations in the molecule have been resolved . Gradient-denaturing gel electrophoresis has the capacity to separate mutant DNA on the basis of the nature and position of the mutation. Basic Life Sci, 1986, 38, 311 - 8 Molecular approaches to the study of nucleotide excision repair in eukaryotes; Friedberg EC et al.; Very little is known about the molecular mechanism of nucleotide excision repair in eukaryotes . Studies on human cells have been stimulated by the availability of excision repair-defective cell lines from patients suffering from the autosomal recessive disease xeroderma pigmentosum (XP) . Such studies have contributed significantly to an understanding of the genetic complexity of excision repair in human cells . However, to date, no human excision repair genes or gene products known to complement the repair defect in XP cells have been isolated . The yeast Saccharomyces cerevisiae is an interesting model for exploring the molecular mechanism of nucleotide excision repair in eukaryotic cells . As is true in human cells, multiple yeast genes are involved and at least five genes are required for the specific incision of UV-irradiated DNA in vivo . These five genes have been isolated by molecular cloning and the nucleotide sequences of four of them have been determined . Each of these cloned genes is being used for overexpression of protein. Am J Pediatr Hematol Oncol, 1986 Summer, 8(2), 99 - 104 Antibody response to Escherichia coli L-asparaginase . Prognostic significance and clinical utility of antibody measurement; Cheung NK et al.; By using a modification of the microtiter solid-phase radioimmunoassay, we have measured Escherichia coli L-asparaginase (L-ASP) specific IgG, IgG4, and IgE antibodies in children who received L-ASP as part of their chemotherapy for leukemia and lymphoma . In 13 children with acute lymphoblastic leukemia induced with vincristine, prednisone, and L-ASP (10,000 IU/M2 i.v . each week for 3 weeks), seven developed high titer specific IgG antibodies . Four of the seven relapsed at the time of their peaking IgG response (6-10 months) . None of the six with low or absent L-ASP antibody response have relapsed (followed for 20-35 months) . In six children with allergic reactions to L-ASP reinduction, all had high titers of L-ASP specific IgG4 (greater than or equal to 20 U/ml) at the time of their reaction . In 16 other children with low L-ASP IgG4 (less than 13 U/ml), none demonstrated allergic reactions to rechallenge . Specific IgE was not consistently detectable in either group . In 21 patients with leukemia or lymphoma on L-ASP with cyclophosphamide-containing regimens, none developed significant IgG antibody response, compared with seven of 13 not receiving cyclophosphamide (p less than 0.001) . We conclude: (a) development of L-ASP antibodies may have prognostic significance; (b) the detection of specific IgG4 can predict L-ASP allergy; and (c) cyclophosphamide-containing regimens reduce antibody formation to L-ASP and may allow repetitive (without anaphylaxis) and more effective (avoiding neutralizing antibodies) use of L-ASP. Vet Med Nauki, 1986, 23(4), 7 - 11 {Isolation of Escherichia coli O 157 from pigs with diarrhea}; Petkov A et al.; Biochemical and serologic studies were carried out with Escherichia coli strains isolated from pigs affected with diarrhea . A total of 161 were investigated, 13 (8.01 per cent) of which were found to belong to serogroup O 157 . Almost all strains proved beta-hemolytic, enterotoxigenic, and K 88-positive . The strains of this serogroup were found for the first time in this country in swine colibacteriosis. Med Microbiol Immunol (Berl), 1986, 175(4), 251 - 60 Indirect haemagglutination test for the detection of thermolabile (LT) enterotoxin from Escherichia coli; Ricci LC et al.; An indirect haemagglutination test (IH) for the detection of enterotoxigenic E . coli (LT) was developed . Twenty-five enterotoxigenic E . coli (ETEC) from human and porcine diarrhoea and from river water were examined . The described IH test was more specific and sensitive than the passive immune haemolysis test (PIH). Med Microbiol Immunol (Berl), 1986, 175(4), 221 - 7 Treatment of LD100 Escherichia coli septic shock with netilmicin and methylprednisolone in baboons; Flournoy DJ et al.; Treatment efficacy with netilmicin sulphate/methylprednisolone sodium succinate in a severe septic shock baboon model, using an LD100 of live Escherichia coli, was evaluated . All the animals treated with both netilmicin and methylprednisolone were permanent (greater than or equal to 7 days) survivors, whereas none of the untreated baboons lived more than 24 hours . These results indicate that, in a baboon model, netilmicin is an effective alternative to gentamicin (with methylprednisolone) in the treatment of severe septic shock. Int J Immunopharmacol, 1986, 8(3), 357 - 68 Recombinant human tumor necrosis factor--II . Antitumor effect on murine and human tumors transplanted in mice; Sohmura Y et al.; Recombinant human tumor necrosis factor (rHu-TNF) was found to exhibit potent antitumor activities not only against murine tumors, i.e . Meth A sarcoma, B 16 melanoma, colon 26 adenocarcinoma, Lewis lung carcinoma and MH134 hepatoma, transplanted in syngeneic mice but also against human tumors, i.e . HMV-2 melanoma, PC-10 lung carcinoma and GOTO neuroblastoma, heterotransplanted in nude mice . rHu-TNF caused necrosis of all tumors tested and inhibited their growth in a dose dependent manner . Complete regression of tumors was observed in mice bearing Meth A, B16, colon 26, MH134, HMV-2 and PC-10 but not in mice bearing Lewis lung carcinoma and GOTO neuroblastoma . The prolongation of survival time was also observed in syngeneic mice transplanted with murine tumors except Lewis lung carcinoma . The antitumor effect of rHu-TNF was more evident when it was given intratumorally than when given intravenously . The feasibility of rHu-TNF as a drug for cancer therapy is discussed. Int J Immunopharmacol, 1986, 8(3), 347 - 55 Recombinant human tumor necrosis factor--I . Cytotoxic activity in vitro; Nakano K et al.; Cytotoxic activity of recombinant human TNF (rHu-TNF) on various human cell lines was examined in vitro . rHu-TNF exerted a cytostatic effect on various types of human tumor cells such as carcinoma, sarcoma, leukemia, melanoma and other types . When the cytocidal effect was examined on the tumor cells which were cytostatically susceptible to rHu-TNF, the cytocidal effect of rHu-TNF was also noticed on many of these tumor cells . However, some tumor cells were affected cytostatically only . Human diploid cells were not affected cytostatically or cytocidally by rHu-TNF . WI-38 VA13 cells which are an SV-40-transformed derivative of WI-38 diploid cells, were affected both cytostatically and cytocidally by rHu-TNF . These results suggest that rHu-TNF exerts cytostatic and cytocidal effects against a broad spectrum of human tumor cells, and its cytotoxic activity is tumor-specific. Int J Immunopharmacol, 1986, 8(3), 313 - 21 A comparative evaluation of particulate and soluble glucan in an endotoxin model; Bowers GJ et al.; Particulate glucan (P) but not soluble glucan (F) has been shown to sensitize rats to endotoxins . This phenomenon is believed to be mediated by the reticuloendothelial system (RES) . The effect of glucan-P and -F on the RES, and the response of glucan-treated rats to nonlethal doses of endotoxin were investigated . Rats were injected for 5 days with 10 mg/kg of glucan-P, -F or saline . Three days later rats were either (1) injected with colloidal carbon for clearance studies, (2) sacrificed for organ histology and determination of serum glucose, plasma thromboxane (Tx) B2, and plasma 6-keto-prostaglandin (PG) F1 alpha concentrations, or (3) challenged with a nonlethal dose of endotoxin . The latter were further subdivided into groups for either 30-day survival or for sacrifice at 30 min or 4 h post-endotoxin infusion to obtain blood samples for glucose, TxB2, and 6-keto-PGE1 alpha determinations . Glucan-P induced hepatosplenomegaly and granulomatous changes within the liver and spleen . The carbon clearance halftime was markedly decreased in these animals . In glucan-P-treated rats challenged with endotoxin, elevated concentrations of both plasma prostanoids were observed as well as alterations in serum glucose levels . These changes were less pronounced in glucan-F- or saline- treated rats . Following endotoxin challenge, only 40% of glucan-P-treated rats survived 30 days whereas 100% of both the glucan-F and saline-treated rats survived . We conclude that glucan-P, in contrast to glucan-F, significantly heightens RES function and that this effect likely accounts for the endotoxin sensitivity. Environ Mutagen, 1986, 8(4), 571 - 7 Induction of SOS genes of Escherichia coli by chromium compounds; Llagostera M et al.; The induction of several SOS genes of Escherichia coli such as recA, umuC, and sfiA by hexavalent (K2Cr2O7, K2CrO4, and CrO3) and trivalent (CrCl3, Cr(NO3)3, and (CH3COO)3Cr) compounds of chromium was studied . Induction was measured as beta-galactosidase activity, using lacZ gene fusions under the control region of different SOS genes . The hexavalent chromium forms induced the genes responsible for massive synthesis of RecA protein, error-prone repair, and inhibition of cell division . On the other hand, the trivalent chromium compounds were unable to induce any of the SOS genes tested . Individual assay of hexavalent chromium compounds showed that K2Cr2O7 was a stronger inducing agent of those three SOS genes tested than K2CrO4, which, in turn, was stronger than CrO3 . All this data led to the conclusion that hexavalent chromium compounds, but not trivalent, are proficient agents of induction of the SOS system and can produce indirect mutagenesis in Escherichia coli. CRC Crit Rev Biochem, 1986, 21(1), 27 - 52 Molecular biology of terminal transferase; Chang LM et al.; Terminal transferase is an unusual deoxynucleotide polymerizing enzyme found only in prelymphocytes . The protein was purified to homogeneity from calf thymus glands in 1971 as a 32 kDa protein with a two peptide structure . Subsequent biochemical and immunological analyses of terminal transferase protein in crude extracts from a number of animal species showed a single peptide with a molecular weight of about 58,000 . The two peptide structure found earlier was caused by proteolysis . Homogeneous 58 kDa terminal transferase has now been produced from human lymphoblastoid cells and calf thymus glands by immunoaffinity chromatography . In vitro phosphorylation studies showed that the terminal transferase protein contains one phosphorylation site near one end of the polypeptide chain, and the phosphorylation of the enzyme has been confirmed by in vivo labeling experiments . Unambiguous demonstration of the molecular weight of the human terminal transferase was obtained by translation of the cloned human terminal transferase DNA sequence to a 58,308 Da protein . The translated amino acid sequence also provided a possible phosphorylation site near the amino-terminus of the protein . Preliminary analysis of the genomic structure shows a simple intron/exon pattern with the total human terminal transferase gene spanning at least 65 Kb. Ann N Y Acad Sci, 1986, 469, 97 - 103 Model for the dynamics of colicin plasmids in continuous culture; Lauffenburger DA; Bacteriocin plasmids may be useful in preventing plasmid instability because the bacteriocin they produce selectively kills otherwise identical bacterial cells lacking the plasmid . This capability may allow plasmid-bearing cells to persist despite the problems of growth-rate depression and segregational plasmid loss that often lead to displacement by plasmid-free cells . In order to determine the conditions under which bacteriocin plasmids can prevent instability, we have developed and analyzed a mathematical model for the growth of colicin plasmid-bearing E . coli in continuous culture . Model behavior compares successfully with experimental results of Adams et al . Dependence of the system dynamic behavior on key parameters has been elucidated and a simple parameter criterion for prevention of plasmid instability has been derived . The possibility of instability prevention was found to be extremely sensitive to dilution rate. Vet Med Nauki, 1986, 23(1), 3 - 9 {Therapeutic effect and residues of a Gentavet preparation in chicks, piglets and calves}; Vangelov S et al.; Studied were the therapeutic and prophylactic effects of the preparation Gentavet (containing gentamycin complex II) with pigs, birds and calves affected with colibacteriosis . It was found that in conc . 80,000 IU (0.08 g or 0.8 g) with the drinking water (1 lit.) given 14 days to birds lowered the mortality rate to a higher extent than Avimycin (containing oxitetracycline and neomycin) . At the rate of 10,000 IU (0.01 g or 0.1 g) per kg body mass, orally, split in two for 24 h with newborn pigs Gentavet produced a very good prophylactic effect, and when used at a higher rate--16,000 IU (0.016 g or 0.16 g) per kg under the same conditions up to 90 per cent of the pigs with polyenteritis recovered . At 10,000 IU (0.01 g or 0.1 g) per kg body mass, orally, split in two for 24 h with newborn calves with colibacteriosis produced therapeutic and prophylactic effects . Gentamycin complex II residual amounts remained longest in the parenchymal tissue of the kidneys and liver . The withdrawal times in slaughtering birds, pigs, and calves treated with Gentavet should last from 30 to 46 days (31 days for birds, 46 days for pigs and calves of up to 100 kg, and 30 days for heifers of up to 400-500 kg) . This period could be shortened 4 to 5 days if both kidneys and liver are to be eliminated after slaughter. Gene, 1986, 42(1), 49 - 57 High-level expression vectors to synthesize unfused proteins in Escherichia coli; Seth A et al.; A new class of plasmid vectors (pANK-12, pANH-1, and pPL2) for synthesizing unfused proteins was constructed by inserting synthetic linkers at the NdeI site (CATATG) of plasmid pJL6, which contains the lambda cII gene initiator codon . These expression vectors contain the lambda pL promoter, the cII ribosome-binding site, cII start codon and unique restriction sites (KpnI, Asp718, HpaI, BamHI) downstream from the initiator ATG for expression of unfused proteins . The main advantage of these vectors is that any DNA fragment with an open reading frame that does not possess a start and/or a stop codon can be directed to overproduce protein in an unfused form. Circ Shock, 1986, 19(1), 55 - 67 Metabolic effects of sodium dichloroacetate in endotoxemic minipigs; Weingand KW et al.; Lactic acidosis is significant in the pathophysiology of endotoxicosis . By increasing the activity of pyruvate dehydrogenase, sodium dichloroacetate (DCA) decreases lactate production and may be useful as a therapy for endotoxic shock . Five (n = 5) 50-80-kg Yucatan minipigs were fitted with jugular, portal, hepatic vein, and carotid artery catheters, and hepatic artery and portal vein flow cuffs to quantitate transhepatic kinetics of glucose, lactate, and insulin . Three days later, they were placed in slings, and a primed-continuous intravenous infusion of 3-tritiated glucose was initiated to monitor whole body glucose kinetics . Following a 3-hour control period, an intravenous infusion of E . coli endotoxin (LPS) was administered at 15 micrograms/kg/hr for 6 hours . After 1 hour of LPS infusion, DCA was administered as a primed (30 mg/kg)-continuous (30 mg/kg/hr) intravenous infusion for 5 hours . These DCA-treated endotoxemic minipigs (group DE) were compared statistically to a group (n = 8) of minipigs given endotoxin only (group E) . Group DE arterial lactate concentrations eventually decreased (19.2 +/- 1.0 mg/dl) significantly (P less than or equal to 0.05) below group E (39.3 +/- 7.7 mg/dl) in the last hour of DCA infusion . The progressive hypoglycemia of group DE decreased below that of group E and became significantly (P less than or equal to 0.05) lower in the last hour of the experiment (33.6 +/- 11.5 vs 50.6 +/- 10.0 mg/dl, respectively) due to decreased hepatic gluconeogenesis as evidenced by a lower relative rate of appearance (%Ra) of glucose and decreased hepatic glucose output and lactate extraction . A large relative decrease in pancreatic output of insulin in group DE contributed to the lower serum insulin levels than group E throughout the treatment period . Lethality in group DE (60%) was unaltered from that of group E (67%) . Despite its ability to decrease arterial lactate concentrations, DCA alone does not appear to be an effective treatment for endotoxicosis. Circ Shock, 1986, 19(1), 39 - 45 Potentiated cardiodepressant effect of serum by endotoxin in adrenalectomized rats; Carli A et al.; We have previously shown that in intact rats (IT) a sublethal and nonhypotensive dose (2 mg/kg IV) of Escherichia coli endotoxin was able to induce the early and sustained release of a lipid-soluble cardiodepressant factor, decreasing contractility of cultured rat heart cells by about 35% . As a humoral mediation may also be envisaged in cardiac dysfunction observed by other investigators in adrenal insufficiency, this study was designed to assess serum cardiodepressant effects of endotoxin in 6-10-day, saline-maintained, adrenalectomized rats (ADX) . In ADX 2 mg/kg endotoxin caused severe hypotension and 100% lethality within the first 90 min . To obtain in ADX a depressant effect of serum on cultured heart cells fairly similar to that observed in IT, it was necessary to reduce the endotoxin dose by about 200 times (0.01 mg/kg) . The latter proved sublethal and nonhypotensive in ADX and was without depressant effect of serum in IT . Serum from ADX (no endotoxin) induced a slight but significant decrease by about 9% in cultured heart cell contractility when compared to serum from IT or sham-operated controls . These data show that adrenalectomy confers a cardiodepressant effect on rat serum and potentiates the release of endotoxin-induced cardiodepressant substance(s) without relation to systemic hypotension. Ultramicroscopy, 1986, 19(1), 43 - 56 Contrast and resolution of scanning transmission electron microscope imaging modes; Reichelt R et al.; Image blurring due to delocalization of inelastic events was studied for scanning transmission electron microscopy (STEM) of unstained thin sections . The delocalization probability was obtained from the angular distribution of inelastic scattering, which was calculated from experimental electron loss spectra of organic samples . This probability was implemented in a Monte Carlo program to simulate the effects of multiple scattering and delocalization for STEM images collected by either the annular detector or the spectrometer, and images generated by a combination of these two signals . Depending on the illumination, the detector geometry and the energy-loss range selected for imaging the annular detector image is blurred by a non-negligible fraction of inelastically scattered electrons . Simultaneous acquisition of an inelastic image using a spectrometer allows the blurring to be reduced by calculation of either the ratio or the difference of the two darkfield signals . While inherent nonlinearities reduce the interpretability of ratio-contrast images, difference-contrast improves the visibility of details submerged in a diffuse background without introducing artifacts. Proc Natl Sci Counc Repub China B, 1986 Jan, 10(1), 35 - 42 Lung vascular injury after Escherichia coli endotoxin and phorbol myristate acetate infusion in dogs; Hsu K et al.; The effects of Escherichia coli endotoxin and phorbol myristate acetate (PMA), a potential stimulator of polymorphonuclear leukocyte (PMN), on circulating PMN counts, gas exchange, protein concentration of lavage fluid, pulmonary hemodynamics and pathology of the lung were studied in ten anesthetized dogs . Six dogs were infused with 1 microgram/kg endotoxin plus 10 micrograms/kg of PMA; four other dogs were infused with the same amount of endotoxin but 5 micrograms/kg of PMA . After administration of endotoxin plus 10 micrograms/kg PMA, the number of circulating PMN (per mm3) decreased dramatically from 4081 +/- 1041 to 303 +/- 119, arterial oxygen partial pressure (PaO2) dropped to 49.1 +/- 2.4 mmHg and the arterial alveolar oxygen partial pressure difference (A-a DO2) increased significantly above baseline . Lungs from this group appeared to be grossly damaged: edema with distinct petechial hemorrhage and areas of hemorrhagic consolidation; frothy edema fluid often emanated from the tracheas . The group infused with endotoxin plus 5 micrograms/kg PMA showed no significant decrease in the number of PMN; PaO2 and A-a DO2 maintained comparatively stable . Protein concentration of lavage fluid and lung wet/dry weight ratios in dogs of 10 micrograms/kg PMA group were significantly increased (P less than 0.05) as compared to those of 5 micrograms/kg PMA group . Our study showed that the magnitude of leukopenia after endotoxin and PMA was paralleled with the severity of lung vascular injury . These results support the potential role of PMN in the pathogenesis of acute edematous lung injury. Physiol Behav, 1986, 36(4), 745 - 9 Effect of centrally administered interleukin-1 and endotoxin on food intake of fasted rats; McCarthy DO et al.; We have previously shown that interleukin-1 (IL-1), a polypeptide known to mediate many aspects of the acute phase response to infection, suppresses food intake when injected intraperitoneally into fasted rats . IL-1 acts at the level of the hypothalamus to induce fever . In view of the large number of peptides that have been shown to alter food intake as well as body temperature when injected intracerebroventricularly (ICV), we hypothesized that the receptor site for the anorexigenic activity of IL-1 would be located in a central nervous site bathed by the cerebrospinal fluid . In the present study, ICV injection of IL-1 or E . coli endotoxin (a stimulus for the synthesis of IL-1), significantly elevated body temperature, but did not affect food intake of fasted rats . We conclude that receptors mediating the anorexigenic actions of IL-1 or endotoxin are not located at a central nervous site bathed by the cerebrospinal fluid . Furthermore, fever per se is not responsible for the reduction in food intake seen following peripheral injection of IL-1 or endotoxin. J Gen Microbiol, 1986 Jan, 132 ( Pt 1), 171 - 81 Effects of pho regulatory mutations and phoA gene amplification on alkaline phosphatase synthesis and release by lky mutants of Escherichia coli K12; Atlan D et al.; lky mutants of Escherichia coli K12 spontaneously released alkaline phosphatase (APase) into the extracellular medium to give up to 300 units ml-1 . APase is a phosphate repressible periplasmic enzyme encoded by the gene phoA . With a view to establishing a method of easy purification, we have analysed APase synthesis and release patterns of isogenic lky strains containing either a constitutive pho regulatory mutation, or a hybrid plasmid carrying the structural gene phoA+ and pho regulatory genes, or a transducing phi 80 phoA+ phage . In the presence of the phoS2333 mutation, F- lky strains lysogenized with phi 80 phoBin phoA+ phage and grown in high phosphate medium were able to release eight times more APase activity (2300 units ml-1) than haploid strain 2336 (phoS+ lky) grown in low phosphate medium . Neither protein synthesis, the cell export machinery nor leakage mechanisms were limiting for APase release . Sufficient APase was released into the medium to facilitate its purification. Int J Nucl Med Biol, 1986, 12(6), 477 - 81 {Detection of endotoxins in radiopharmaceutical preparations--III . Limulus test assessment using radiopharmaceutical preparations; correlation with the rabbit pyrogen test}; Cohen Y et al.; Experiments using 17 radiopharmaceuticals containing known amounts of added endotoxin show that none of them inhibits the pyrogenic reaction of the rabbit . Gelation of the Limulus amoebocyte lysate (LAL) is inhibited by 4 of them: colloidal erbium 169Er citrate, colloidal rhenium 186Re sulfide, colloidal technetium 99mTc (Re) sulfide for liver scintigraphy and the colloidal technetium 99mTc (Re) sulfide for lymphography . This inhibition is cancelled, either by dilution or after neutral pH adjustment . Both controls were performed on 313 batches of various radiopharmaceuticals, 95% of results were identical (93% negative, 2% positive) . The remaining 5% correspond to positive LAL tests vs negative rabbit tests on the same batches . No negative LAL test vs positive rabbit test was observed. Int J Nucl Med Biol, 1986, 12(6), 471 - 6 {Detection of endotoxins in radiopharmaceutical preparations--II . Comparison of the sensitivity of methods using the rabbit and the Limulus amoebocyte lysate for the detection of endotoxins}; Bruneau J et al.; The rise of the rabbit internal temperature after i.v . injection of an endotoxin solution is proportional to concentration . Gelation of Limulus amoebocyte, when in presence of an endotoxin solution, is also related to concentration . We compared the sensitivity of these two methods . With our experimental procedure, the rabbit is sensitive to a 0.40 ng/mL solution and the Limulus amoebocyte lysate to a 0.14 ng/mL solution . The rabbit sensitivity increase is related to the per kilogramme injected volume, whereas sensitivity is not related to the volume to check in the case of the lysate. Int J Nucl Med Biol, 1986, 12(6), 467 - 70 {Detection of endotoxins in radiopharmaceutical preparations--I . Comparison of rabbit hyperthermia after intravenous or intrathecal administration of reference endotoxin preparations}; Merlin L et al.; The rise of the rabbit internal temperature after endotoxin injection is related to the route of administration . A rise of 1.71 +/- 0.411 degrees C is obtained after i.v . injection of 1 ng/kg Escherichia coli 0111.B.4 endotoxin . An increase of 1.93 +/- 0.236 degrees C is obtained after suboccipital intrathecal injection of 0.1 ng/kg of the same endotoxin; with the intrathecal route, the hyperthermia is induced by E . coli endotoxin after a dose ten times lower than with i.v . injection as shown by statistical analysis. Folia Microbiol (Praha), 1986, 31(2), 81 - 5 The control of adenylate cyclase activity in Escherichia coli; Janecek J et al.; Cell-free extract of E . coli possessed an inhibited adenylate cyclase activity after a previous anaerobic incubation of cells with glucose which is transported and metabolized . The degree of the inhibition depends on incubation conditions . Glucose analogues that are only transported but not metabolized, are not inhibitory . To restore the adenylate cyclase activity, the cells have to be cultivated aerobically prior to disintegration for a defined period of time without glucose. Gene, 1986, 41(2-3), 225 - 31 Oligonucleotide directed mutagenesis of cauliflower mosaic virus DNA using a repair-resistant nucleoside analogue: identification of an agnogene initiation codon; Dixon L et al.; Mutation of the initiation codon of the dispensible open reading frame, ORF VII, of cauliflower mosaic virus (CaMV) delayed the appearance of disease symptoms, but the mutants reverted with high frequency . This suggests a role of this start codon in viral expression . Oligonucleotide-directed mutagenesis, utilizing a novel, repair-resistant deoxyguanosine analogue, 2'-deoxy-7-deazainosine (dDI), highly improved the yield of mutants. Int J Biochem, 1986, 18(5), 431 - 5 Uridine phosphorylase from Escherichia coli B . Enzymatic and molecular properties; Vita A et al.; Uridine phosphorylase (EC 2.4.2.3) from Escherichia coli B is an oligomeric protein composed of four identical subunits of 29,000 mol . wt . The enzyme has four half-cystine residues per subunit titrable only in denaturing condition . No disulphide linkages either inter- or intra-chain are present . The isoelectric point is 5.25 . The enzyme shows strict specificity toward uridine and 5-methyluridine and is inhibited by thymine, deoxycytidine and heavy metal ions. Eur Surg Res, 1986, 18(2), 112 - 21 Application of a quantitative spectrophotometric endotoxin assay on lymph and plasma from the rat; Olofsson P; A quantitative spectrophotometric assay for endotoxin, utilizing Limulus amebocyte lysate and a chromogenic peptide substrate, was standardized for the application on lymph and plasma from the rat . The influence of inhibitors and unspecific amidolytic activity, the optimal incubation times, the adhesion of endotoxin to platelets and glass surface, the variation in endotoxin standard as well as the effects of cold storage on endotoxin activity were settled . In vitro added as well as endogenously derived endotoxin was studied . The method was found to be reliable when in practical use on these media. Res Vet Sci, 1986 Jan, 40(1), 128 - 35 Emigration of polymorphonuclear leucocytes into the intestinal lumen of the neonatal piglet in response to challenge with K88-positive Escherichia coli; Sellwood R et al.; The emigration of neutrophils from the blood of neonatal piglets into the intestinal lumen in response to a K88-positive strain of Escherichia coli was investigated . The pig herd used was of known genetic susceptibility to K88-positive E coli and had recently experienced an outbreak of neonatal diarrhoea . Neutrophil emigration depended on certain factors . Neutrophils emigrated into ligated loops in susceptible piglets sucking immune colostrum from susceptible dams but not into loops in colostrum deprived resistant piglets or piglets sucking non-immune colostrum from resistant dams . In susceptible, colostrum deprived piglets neutrophils in intestinal contents were only associated with severe lesions . Large numbers of neutrophils which appeared at several foci on the villi were observed in three of six resistant piglets that sucked immune colostrum from susceptible dams . It was concluded that neutrophil emigration into the intestinal lumen of piglets could occur in response to K88-positive E coli and resulted not from the presence or absence of the intestinal K88-receptor but from the ingestion of immune colostrum. J Clin Microbiol, 1986 Jan, 23(1), 92 - 9 Immunologic response of patients with legionellosis against major protein-containing antigens of Legionella pneumophila serogroup 1 as shown by immunoblot analysis; Sampson JS et al.; Major protein-containing antigens of Legionella pneumophila serogroup 1 were were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis with rabbit antisera to 14 different Legionella species or serogroups . Fourteen bands were observed in immunoelectropherograms of whole-cell, sonicated cell, and heated cell preparations, seven of which appeared in the supernatant fluid from the heated cells and three of which were shown in an outer membrane fraction . Immunoblots of whole-cell antigen preparations of 14 Legionella species or serogroups revealed seven major Legionella proteins: antigens with molecular weights of 58,000, 79,000, and 154,000 were present in all Legionella sp . strains, antigens with molecular weights of 44,000 and 97,000 occurred in multiple species, and antigens with molecular weights of 14,000 and 25,000 were present only in L . pneumophila strains . All sera from 15 patients with culture-confirmed L . pneumophila serogroup 1 disease and 14 of 18 (78%) sera from serologically diagnosed patients reacted with the 58-kilodalton (kDa) common antigen . In contrast, less than one-half of the sera reacted with the L . pneumophila-specific proteins (14 and 25 kDa) . Absorption of sera with Escherichia coli cells had no effect on their reactivity with the 58-kDa antigen, whereas absorption with L . pneumophila serogroup 1 cells removed reactivity . These data suggest that the 58-kDa antigen may prove useful in serodiagnostic tests for legionellosis. Gene, 1986, 41(1), 67 - 74 Mutations that reduce expression from the P2 promoter of the Escherichia coli galactose operon; Bingham AH et al.; We describe the isolation and characterisation of twelve different mutations that reduce gene expression from the galP2 promoter, starting with a gal regulatory region with a mutation that inactivated galP1, the cAMP-CRP-dependent promoter . Seven of the new mutations reduce the initiation of transcription at P2 whereas the others reduce translation initiation of the first gal operon gene, galE . Two of the mutations affecting translation fall in the galE initiation codon and the Shine-Dalgarno sequence . Mutations that allow the formation of a stem-loop structure in the messenger including this sequence also reduce translation . A deletion of 11 bp, upstream of the Shine-Dalgarno sequence, almost totally prevents translation . Although none of the point mutations that reduced transcription initiation at P2 fall in the -35 region, we repeatedly isolated insertions in this zone . The point mutations all fell around the -10 region: the strongest effects were found with mutations that altered the sequence away from the consensus that has been established for Escherichia coli promoters . The effects of the two strongest P2 mutations were investigated in the absence of the P1 mutation used for their isolation . One mutation, a T:A to C:G transition at -12, inactivates both P2 and P1 . In contrast the other, a T:A to G:C transversion at -19, specifically inactivates P2, but leaves P1 partially active even in the absence of cAMP-CRP . The implications of this are discussed in the context of how cAMP-CRP controls the balance between transcription from P2 and P1 at the gal operon regulatory region. Environ Mutagen, 1986, 8(2), 263 - 71 The genetic toxicology of metal compounds: II . Enhancement of ultraviolet light-induced mutagenesis in Escherichia coli WP2; Rossman TG et al.; Salts of metals which are carcinogenic, noncarcinogenic, or of unknown carcinogenicity were assayed for their abilities to modulate ultraviolet (UV)-induced mutagenesis in Escherichia coli WP2 . In addition to the previously reported comutagenic effect of arsenite, salts of three other compounds were found to enhance UV mutagenesis . CuCl2, MnCl2 (and a small effect by KMnO4), and NaMoO4 acted as comutagens in E coli WP2, which has wild-type DNA repair capability, but were much less comutagenic in the repair deficient strain WP2s (uvrA) . The survival of irradiated or unirradiated cells was not affected by these compounds . No effects on UV mutagenesis were seen for 16 other metal compounds . We suggest that the comutagenic effects might occur either via metal-induced decreases in the fidelity of repair replication or (in the case of CuCl2) via metal-induced depurination. Circ Shock, 1986, 18(4), 289 - 300 Metabolic effects of glucagon in endotoxemic minipigs; Weingand KW et al.; A treatment group of four 50-80-kg Yucatan minipigs was fitted with jugular, portal, hepatic vein, and carotid artery catheters, and hepatic artery and portal vein flow cuffs to quantitate transhepatic kinetics of glucose, lactate, and insulin . Three days later, they were placed in slings, and a primed-continuous intravenous infusion of 3-tritiated glucose was initiated to monitor whole body glucose kinetics . Following a 3-hour control period, an intravenous infusion of E . coli endotoxin was administered at 15 micrograms/kg/hr for 6 hours . After 1 hour of endotoxin infusion, glucagon was administered as a primed (50 micrograms/kg)-continuous (50 micrograms/kg/hr) intravenous infusion for 5 hours . These results were compared statistically to a control group of eight minipigs given endotoxin only . Glucagon caused a transient (less than 1 hour) peak elevation of arterial glucose levels due to increased hepatic glycogenolysis immediately following initiation of the glucagon infusion . Despite enhanced extrahepatic (renal) gluconeogenesis, plasma glucose concentrations decreased to the hypoglycemic levels of the control group, as glucagon was unable to overcome the relative endotoxic inhibition of hepatic gluconeogenesis . Pancreatic output of insulin increased sixfold resulting in threefold increments in transhepatic uptake and arterial serum insulin levels . Arterial lactate and pyruvate concentrations were elevated due to increased peripheral production and hepatic output . Survivability in the treatment group (75%) improved over the control group (33%). Circ Shock, 1986, 18(4), 267 - 75 Regional blood flow during continuous low-dose endotoxin infusion; Fish RE et al.; Escherichia coli endotoxin (ET) was administered to adult rats by continuous IV infusion from a subcutaneously implanted osmotic pump (Alzet) . Cardiac output and regional blood flow were determined by the radiolabeled microsphere method after 6 and 30 hr of ET or saline infusion . Cardiac output (CO) of ET rats was not different from time-matched controls, whereas arterial pressure was 13% lower after 30 hr of infusion . After both 6 and 30 hr of ET, pancreatic blood flow and percentage of cardiac output were lower than in controls . Estimated portal venous flow was decreased at each time point, and an increased hepatic arterial flow (significant after 30 hr) resulted in an unchanged total hepatic blood flow . Blood flow to most other tissues, including epididymal fat, muscle, kidneys, adrenals, and gastrointestinal tract, was similar between treatments . Maintenance of blood flow to metabolically important tissues indicates that the previously reported alterations in in vitro cellular metabolism are not due to tissue hypoperfusion . Earlier observations of in vitro myocardial dysfunction, coexistent with the significant impairment in pancreatic flow, raise the possibility that release of a myocardial depressant factor occurs not only in profound shock but also under less severe conditions of sepsis and endotoxemia. Circ Shock, 1986, 18(3), 179 - 92 Effects of methylprednisolone upon vascular permeability changes in endotoxic shock; Hubbard JD et al.; This study was designed to examine glucocorticoid effects on increases in vascular permeability caused by endotoxic shock in dogs anesthetized with pentobarbital sodium (30 mg/kg) . Methylprednisolone sodium succinate was administered IV in two doses (30 mg/kg each) before Escherichia coli endotoxin was administered (0.5 mg/kg) . Samples of serum and lymph from the left thoracic duct were collected for measurement of total protein and separation by polyacrylamide gel electrophoresis . The same four protein electrophoretic fractions with molecular weights (M.W.) ranging from 60,000 to 1,000,000 were consistently chosen for analysis . Methylprednisolone treatment given prior to endotoxin administration resulted in an attenuation of the early increase in total protein flux and lymph to plasma protein (L/P) ratio and prevented significant increases in the permeability surface area product observed in the group given endotoxin alone . Endotoxin administration alone resulted in significant increases in the permeability-surface area product and L/P ratio for all four electrophoretic fractions . Pretreatment with glucocorticoid partially attenuated the increase in the L/P ratio for only those fractions of 100,000 M.W . or less . These results suggest MP provides partial but not complete protection from increases in vascular permeability during endotoxic shock. Can J Microbiol, 1986 Jan, 32(1), 52 - 5 Site specificity of metallic ion binding in Escherichia coli K-12 lipopolysaccharide; Ferris FG et al.; The site specificity of metallic ion binding in Escherichia coli K-12 lipopolysaccharide was assessed by collecting high-resolution phosphorus nuclear magnetic resonance spectra in the presence of manganese, a paramagnetic divalent cation . This technique revealed high-affinity interactions between the cation and all of the lipopolysaccharide phosphoryl groups . To ascertain whether the carboxyl groups of 2-keto-3-deoxyoctonate contributed to the metal cation binding, lipopolysaccharide was chemically modified using a glycine ethyl ester - carbodiimide reaction . Of the three available carboxyl groups, only one was neutralized by the exogenously added ligand; the others appeared to be cross-linked within the molecule . By analogy, only one carboxyl group should be freely available for binding metallic ions, while the others are probably neutralized by the close proximity of endogenous amino substituents . Although high-resolution phosphorus nuclear magnetic resonance showed that an intermolecular conformational change had occurred after the carboxyl groups were neutralized, titration with manganese revealed no differences in the apparent strength of the interactions between the cation and the phosphoryl groups . Together, these data suggest that the high affinity of lipopolysaccharide for divalent metallic ions can be attributed primarily to the phosphoryl substituents and not free carboxyl groups. Biol Neonate, 1986, 49(2), 102 - 8 Metabolic effects of endotoxin in newborn rabbits; Molnar D et al.; The metabolic and hormonal effects of Escherichia coli endotoxin injected intraperitoneally (IP) or into the cerebral ventricles (ICV) and that of passive hyperthermia were studied in rabbits aged 6-10 days . Irrespective of the route of administration, endotoxin caused a transient rise in blood glucose with a simultaneous rise in plasma insulin . In contrast, only in the IP, but not in the ICV group, the endotoxin resulted in a rise of the free fatty acid and a fall in the ketone body concentrations by the second hour . The blood level of pyruvate, lactate, alanine and glycerol was not altered by endotoxin . No parameter was affected by the injection of saline or passive hyperthermia. Ann Inst Pasteur Immunol, 1986 Jan-Feb, 137C(1), 39 - 50 {Application of the vapor test for the detection and immunologic determination of Escherichia coli heat labile enterotoxin}; Germani Y et al.; The principle of this thin-layer immunoassay (vapour condensation technique or TVAP) is based on the ability of antibodies to absorb firmly to polystyrene surfaces and to retain their reactivity . A condensation pattern consisting of large confluent water drops is noticable when an antibody-antigen reaction takes place . We used this technique to detect and assay the Escherichia coli heat-labile enterotoxin (ETEC LT+) and compared the results of 53 strains (40 positives and 13 negatives) with single radial immune haemolysis, Gm1-ELISA and Vero cell culture tests . With the reagents used, this reaction was specific for a toxin dilution up to 1/14 . As little as 0.025 micrograms/ml of cholera toxin could be detected . The TVAP-test is simple, rapid and cost-effective . It is thus quite suitable for use in diarrhoeal endemic areas. Scand J Infect Dis, 1986, 18(1), 65 - 70 Effective treatment of diarrhoeal dehydration with an oral rehydration solution containing citrate; Ahmed SM et al.; To compare the clinical efficacy of oral rehydration salts (ORS) from effervescent tablets containing citrate with the WHO recommended ORS for the treatment of dehydration due to acute diarrhoea, a randomized clinical trial was carried out in 57 adults and 58 children . These patients had mild or moderate degrees of dehydration and acidosis due to acute watery diarrhoea that was caused by enterotoxigenic Escherichia coli in 43-47% of the cases . Efficacies were compared by measuring oral fluid intake, stool output, gain in body weight, decrease in serum specific gravity and correction of acidosis during treatment . Successful rehydration and maintenance of hydration was achieved in 25 adults and 24 children treated with citrate containing ORS and 25 adults and 24 children treated with WHO ORS . The mean intake of ORS/kg body weight in children receiving WHO ORS was greater (p less than 0.05) and correction of acidosis was faster than the citrate group during the initial 24 h of therapy (p less than 0.05) . By 48 h, however, both groups showed satisfactory and comparable intake of ORS and correction of acidosis . Thus ORS from effervescent tablets containing sodium citrate base is effective for management of diarrhoea in both adults and children and is a convenient stable form of ORS for use in the home and for travelers. Neuroendocrinology, 1986, 42(4), 285 - 8 Indomethacin, an antipyretic drug, prevents the endotoxin-induced and potentiates the hemorrhage-induced oxytocin release into the plasma of the male rat; Kasting NW; Oxytocin (OXY) is a nonapeptide of hypothalamic origin which has defined roles in the female reproductive functions of lactation and labor . However, OXY may have other physiological functions because of its presence in the male and its release in response to stress . Available evidence suggests prostaglandins may stimulate the release of OXY . These experiments sought to determine if the stressors, endotoxin and hemorrhage, would release OXY in the chronically catheterized, freely behaving male rat and what effect the antipyretic and prostaglandin synthesis inhibiting drug, indomethacin, would have on these responses . Endotoxin caused a marked release of OXY from mean baseline levels of 5 pg/ml to mean peak levels of 168 pg/ml . Indomethacin greatly attenuated this increase . In contrast, OXY release in response to hemorrhage of either 22 or 44% of the blood volume of the rat was enhanced by indomethacin . Indomethacin increased the hemorrhage-induced OXY levels about 2-fold over a 2-hour posthemorrhage period . Indomethacin alone had no effect on OXY levels . These data verify that stress is a potent stimulus for OXY release and strengthen the hypothesis that prostaglandins mediate OXY release . The paradoxical effects of indomethacin on OXY release suggest that the prostaglandins may have different effects on OXY release depending upon the evoking stimulus. Med Microbiol Immunol (Berl), 1986, 175(1), 55 - 60 Stability of thermolabile (LT) enterotoxin produced from enterotoxigenic Escherichia coli strains maintained in vitro; Gatti MS et al.; The stability of thermolabile (LT) enterotoxin in 26 strains of porcine enterotoxigenic Escherichia coli (PETEC) belonging to serogroups 08 and 0149 was assayed by the passive immune hemolysis (PIH) test, over a period of 9 months at -70 degrees C . It was found that the percentage of LT+ colonies (% LT+) and the mean value of hemoglobin release (XHb), could predict a change from LT+ to LT-. Med Microbiol Immunol (Berl), 1986, 175(1), 15 - 26 Phenotypic variations among enterotoxigenic O-groups of Escherichia coli from various human populations; Kuhn I et al.; ETEC isolates from various sources (children from Ethiopia and some Asian countries, and Swedish tourists suffering from traveller's disease) were analysed with the aid of "biochemical fingerprinting", which is a highly discriminative, computerized method designed to subdivide E . coli isolates into different phenotypes . Isolates belonging to the most common ETEC O-groups and others which had not been typeable with available O-antisera were selected . It was found that certain phenotypes of O-groups 6 and 114 could be found in materials from several continents . Phenotypes of other O-groups were usually more restricted to certain geographic areas . Among children in Addis Abeba, 19 out of 25 isolates carrying O-antigen 78 belonged to the same phenotype . Some possible explanations for the fact that certain phenotypes of enterotoxigenic E . coli could be found over the whole world are that they might represent relatively recent developed clones, or they may represent unusually stable clones . Of the isolates that had been nontypeable with available antisera, some had lost their LT-productivity after one year of storage . These isolates proved to belong to a wide variety of phenotypes, whereas nontypeable isolates which were stable LT-producers could be clustered into distinct groups . It is suggested that the non-stable LT-producers are members of the normal E . coli flora of these children, which have occasionally picked up the enterotoxin-producing plasmids, whereas most stable LT-producers represent true ETEC clones. Mol Gen Genet, 1986 Jan, 202(1), 90 - 5 Base pair substitution and frameshift mutagenesis induced by apurinic sites and two fluorene derivatives in a recA441 lexA (Def) strain; Granger-Schnarr M; One of the consequences of the induction of the Escherichia coli SOS system is the increased ability of the cells to perform mutagenesis . Induction of the SOS system is the result of derepression of a set of genes through a regulatory mechanism controlled by LexA and RecA . In response to an inducing signal, RecA is activated in a form that facilitates the proteolytic cleavage of LexA repressor . Previous works have shown that activated RecA plays a second role, i.e . it is required for the establishment of base pair substitution mutations promoted by UV irradiation . Using a forward mutational assay and recA441 lexA (Def) host bacteria, we show that the result can be extended not only to other mutagens promoting base pair substitution mutations (Apurinic sites, Ap sites and N-hydroxy-N-2-aminofluorene, N-OH-AF) but also mutagens promoting frameshift mutations (N-Acetoxy-N-2-acetylaminofluorene, N-AcO-AAF) . In the recA441 lexA (Def) strain all the genes which are part of the lexA regulon, including recA itself, are expressed constitutively . The recA441 mutation allows RecA to acquire its activated form when the bacteria are grown at 42 degrees C . We show that in such strains Ap sites or N-OH-AF induce a high level of mutations only when the bacteria are grown at 42 degrees C . On the other hand, we show that N-AcO-AAF can promote mutations even at 30 degrees C; the number of mutations being increased when the bacteria were grown at 42 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS) Mol Gen Genet, 1986 Jan, 202(1), 162 - 8 Mutational specificity of a proof-reading defective Escherichia coli dnaQ49 mutator; Piechocki R et al.; The dnaQ (mutD) gene product which encodes the epsilon-subunit of the DNA polymerase III holoenzyme has a central role in controlling the fidelity of DNA replication because both mutD5 and dnaQ49 mutations severely decrease the 3'-5' exonucleolytic editing capacity . It is shown in this paper that more than 95% of all dnaQ49-induced base pair substitutions are transversions of the types G:C-T:A and A:T-T:A . Not only is this unusual mutational specificity precisely that observed recently for a number of potent carcinogens such as benzo(a) pyrene diolepoxide (BPDE) and aflatoxin B1 (AFB1), which are dependent on the SOS system to mutagenize bacteria, but it is also seen for the constitutively expressed SOS mutator activity in E . coli tif-1 strains as well as for the SOS mutator activity mediated gap filling of apurinic sites . Because the G:C-T:A and A:T-T:A transversions can either result from the insertion of an adenine across from apurinic sites or arise due to the incorporation of syn-adenine opposite a purine base, we postulate that the DNA polymerase III holoenzyme also has a reduced discrimination ability in a dnaQ49 background . The introduction of a lexA (Ind-) allele, which prevents the expression of SOS functions, led to a significant reduction in the dnaQ49-caused mutator effect . Both, the mutational specificity observed and the partial lexA+ dependence of the mutator effect provoke a reanalysis of the hypothesis that the DNA polymerase III holoenzyme can be converted into the postulated but until now unidentified SOS polymerase. J Biochem (Tokyo), 1986 Jan, 99(1), 303 - 6 Differential effect of phosphatidylethanolamine molecular species on glycerophosphate acyltransferase activity of Escherichia coli; Ishinaga M; The effect of phosphatidylethanolamine (PE) molecular species on the reconstitution of partially purified glycerophosphate acyltransferase of Escherichia coli was investigated . The acyltransferase activity was abolished by 1,2-di-unsaturated (U-U) PE, but not by 1-saturated-2-unsaturated (S-U) PE or 1-saturated-2-cyclopropanoyl PE . Since both the U-U and S-U PE used in the present work are in a fluid state at temperatures above about 30 degrees C, the differential effect cannot be accounted for in terms of the membrane fluidity . Therefore, the inactivation of the reconstituted enzyme was attributed to the large amount of the 1,2-di-cis-vaccenoyl species of PE. Infection, 1986 Jan-Feb, 14(1), 7 - 12 Colonization and persistence of Escherichia coli phenotypes in the intestines of children aged 0 to 18 months; Kuhn I et al.; The aim of the present investigation was to study the intestinal colonization of Escherichia coli in newborn children, and to determine which strains become residential within the human intestine . The E . coli flora of 89 newborn children was studied by repeated sampling during their first 11 or 18 months of life . The E . coli isolates from the samples were subdivided into phenotypes by the aid of biochemical fingerprinting, a method which measures the kinetics of 24 selected biochemical tests as a tool for discriminating bacterial strains . It was found that E . coli strains colonizing children soon after birth persisted longer than strains colonizing them later . Especially those phenotypes which were defined as hospital strains persisted longer . Certain phenotypes were commonly found among the children, and these phenotypes were more persistent and more homogeneous than other phenotypes with respect to their pattern of biochemical activities . They might be specially adapted to colonize the human intestine . It was concluded that the generally long persistence of the first E . coli strains colonizing a newborn child indicates that the first case of bacterial colonization in children may be an event too important to be allowed to happen at random. Int J Biochem, 1986, 18(3), 257 - 62 The synthesis and use of oligodeoxynucleotides in plasmid DNA sequencing; Ner SS et al.; A convenient procedure for the synthesis and purification of oligonucleotides is described . 16-base long primers synthesised by this method were used to investigate DNA sequencing using plasmid DNA as a template . This allowed the further analysis of the E . coli glt A sequence coding for citrate synthase and enabled determination of the 5'-non-coding regulatory region of the aminoglycoside phosphotransferase gene. EMBO J, 1986 Jan, 5(1), 175 - 9 Near ultraviolet DNA damage induces the SOS responses in Escherichia coli; Caldeira de Araujo A et al.; The influence of the growth delay induced by near u.v . radiation on the SOS response was monitored by comparing the level of sfiA expression by means of a sfiA::lacZ fusion in both a nuvA+ cell and an isogenic nuvA mutant . The mutant lacks 4-thiouridine in its tRNA and does not exhibit the near u.v.-induced growth delay . Although the two strains exhibit similar sfiA induction levels after 254 nm irradiation, their behaviour is different after illumination with near u.v . light, including solar u.v . Inducibility is 10-20 times higher in the nuvA mutant than in the parent strain . Furthermore, pre-illumination with broad band near u.v . light does not affect the 254 nm-induced sfiA response in the mutant but reduces it by a factor of 3-4 in the parent strain . The kinetics of sfiA induction in near u.v.-illuminated nuvA+ cells, whether treated with 254 nm light or not, is unusual and follows the growth curve: only after 50 min is sfiA derepression observed . It can be concluded that (i) near u.v.-induced DNA lesions are able to trigger the SOS response and (ii) the growth delay effect reduces this response, whether triggered by u.v . or near u.v . light . Hence 4-thiouridine in tRNA acts as a built-in antiphotomutagenic 'device' protecting Escherichia coli cells against mutagenesis and the induction of the SOS response by near u.v . light and sunlight. Diagn Immunol, 1986, 4(1), 17 - 23 ELISA detection of specific functional antibodies in human serum to Escherichia coli, tetanus toxoid, and diphtheria-tetanus toxoids: normal values for IgG, IgA, and IgM; Moen RC et al.; An inexpensive, easily performed enzyme-linked immunosorbent assay (ELISA) was developed to measure specific IgG, IgA, and IgM antibodies to the common antigens Escherichia coli, diphtheria-tetanus toxoid, and tetanus toxoid . Normal values were established . Classical antibody deficiency disease states were confirmed and delineated by these assays . Additionally, several instances were discovered when functional antibody levels were abnormal when the serum immunoglobulin levels were normal . The use of ELISA assays for antibodies to common antigens provides a useful technique to measure and monitor isotype responses of the humoral immune system. Brain Res Bull, 1986 Jan, 16(1), 99 - 105 alpha-Melanocyte-stimulating hormone infused ICV fails to affect body temperature or endotoxin fever in the cat; Rezvani AH et al.; Permanent cannulae for intracerebroventricular (ICV) infusion were implanted bilaterally in cats following stereotaxic procedures . After colonic temperature was recorded for a one-hour baseline, a 300 microliter ICV infusion was given of CSF control vehicle, 1:100 dilution of W3110 E . coli endotoxin (10(8) organisms/ml) or alpha-melanocyte-stimulating hormone (alpha-MSH) in one of seven doses ranging from 50.0 ng to 50.0 micrograms . Whereas ICV E . coli always induced an intense and prolonged fever of rapid onset, alpha-MSH infused similarly was essentially without effect on the deep body temperature of the normothermic cat . When each of the doses of alpha-MSH was infused ICV, either during the rising phase of an E . coli fever or after the febrile response had reached its asymptote, the core temperature of the cat was unaffected . Similarly, a mixture of E . coli combined with alpha-MSH given ICV failed to alter the characteristics of the rapidly developing fever produced in the cat by this endotoxin . On the other hand, either excess Ca++ ions (50 mM) given ICV or the antipyretic drug . Dipyrone, administered systematically during the course of an endotoxin fever effectively attenuated the animal's elevated body temperature . These results demonstrate that alpha-MSH is apparently neither involved in the central mechanisms underlying normal thermoregulation, nor does it act as an endogenous antipyretic in the cat as has been postulated in another species. Biophys J, 1986 Jan, 49(1), 251 - 8 Alignment and merging of electron microscope images of frozen hydrated crystals of the T4 DNA helix destabilizing protein gp32*I; Grant RA et al.; Low dose cryoelectron microscopy has been used to record images and electron diffraction patterns of frozen hydrated crystals of the single-stranded DNA binding protein gp32*I . Fourier transforms from 13 image areas, corresponding to approximately 40,000 unit cells, were aligned by a minimal phase residual search and merged by vector addition in reciprocal space . Phases from the resulting composite transform were combined with amplitudes from electron diffraction patterns to reconstruct the projected mass density of the gp32*I crystal at 8.4 A resolution. Arch Microbiol, 1986 Jan, 143(4), 400 - 2 Role of relA mutation in the survival of amino acid-starved Escherichia coli; Hecker M et al.; Amino acid-starved cells of Escherichia coli relA+, which contain a large number of glycogen particles, are able to survive in phosphate buffer for a longer time period than their relaxed counterparts . With regard to NH4+ starvation differences in the survival of both strains were not found . NH4+ starved cells of E . coli relA are able to synthesize glycogen but amino acid-starved cells of the relA strain are not . We suggest that the synthesis of glycogen triggered by guanosine tetraphosphate during amino acid starvation is responsible for the prolonged viability of the E . coli relA+ strain. Anal Biochem, 1986 Jan, 152(1), 89 - 93 A nitrocellulose filter binding assay for ribonucleotide reductase; Soderman K et al.; Protein B1, one of the two nonidentical subunits of Escherichia coli ribonucleotide reductase, contains two classes of binding sites for nucleoside triphosphates . One class (h-sites), with a high affinity for dATP (KD = 30 nM) regulates the substrate specificity, while the other (l-sites), with a lower affinity for dATP, regulates overall activity . These classes were defined from experiments involving equilibrium dialysis . Here we describe a sensitive alternative method to measure nucleotide binding to ribonucleotide reductases that gave the same results as equilibrium dialysis . The method involves the protein-specific binding of radioactive nucleotides to nitrocellulose filters . We believe that this method will be useful in binding studies with pure reductases from sources other than E . coli, for the characterization of mutants with changed allosteric properties, and as an assay during purification of reductases containing an h-site for dATP. Plasmid, 1986 Jan, 15(1), 35 - 47 Directly repeated, 20-bp sequence of plasmid R1162 DNA is required for replication, expression of incompatibility, and copy-number control; Lin LS et al.; DNA required in cis for the replication of the broad-host-range plasmid R1162 is located on two contiguous HpaII fragments of 210 and 370 bp . The latter of these contains three and one-half, perfectly conserved, 20-bp directly repeated sequences . The significance of these for plasmid replication, incompatibility, and copy-number control was examined by generating deletions into these repeats and testing the properties of the remaining DNA . We conclude from the results that the direct repeats are essential for expression of incompatibility and for the decrease in copy number observed when the directly repeated DNA is cloned into R1162 . Little, if any, additional DNA is required from the ori region for these properties . Moreover, deletions of intermediate size result in an intermediate level of incompatibility, indicating the importance of the periodic structure of the direct repeats . The directly repeated DNA is also required for an active origin of replication, as are additional, nonrepeated sequences adjacent to this DNA . The properties of the direct repeats are discussed with respect to their possible role in the replication of R1162 DNA. Mol Biol (Mosk), 1986 Jan-Feb, 20(1), 242 - 9 {Symmetry of multienzyme complexes}; Gol'dshtein BN et al.; A model for studying the symmetry of stable states arising from polyenzymic complex conformations is proposed . A formal scheme of submolecular structure self-assembly, on which the model is based, enables it not only to limit the class of conformations but in some cases to determine the structure of a complex in an unambigous manner . The model is shown in its application to polyenzymic complexes of dehydrogenases of alpha-keto acids. Mol Biol (Mosk), 1986 Jan-Feb, 20(1), 224 - 32 {A model of an enzyme with mixed cooperation: 3 states of aspartate transcarbamylase}; Saifullin SR et al.; Allosteric enzyme models on the basis of the known properties of aspartate transcarbamylase (ATCase) from Escherichia coli are suggested . In the first model molecules are supposed to equilibrate between two states . In contrast to the classical Monod-Wyman-Changeux model the symmetry of enzyme molecules changes during the conformational transition . It is shown that the number of binding sites of the enzyme defined from the Scatchard plots is sufficiently dependent on values of parameters of enzyme reaction . This fact results from the mixed (both positive and negative) cooperative effects . However the complex kinetic of ATCase is not completely simulated by this model . Therefore the model is complicated by taking into account the inactive third state of the enzyme . Thus the complex kinetic behaviour of ATCase is explained . The models may be also used for other enzymes. Microbiologica, 1986 Jan, 9(1), 53 - 61 Toxicity of granular activated carbon treated coal gasification water as determined by the Microtox test and Escherichia coli; Makino Y et al.; The Microtox assay and various parameters (growth, ATP concentration and electrochemical detection) of Escherichia coli were used to assess the toxicity of various levels of granular activated carbon treated coal gasification process water . The generation time of E . coli was statistically significantly slower at the level of 50 percent treatment than any other level of treatment . No differences were seen for ATP concentration per cell or in the electrochemical detection methods for any level treatment . There was a very high correlation between total organic carbon removal by GAC treatment and reduction in toxicity as measured by the Microtox system . However, even the treated water which had 91 percent of the TOC removed was still highly toxic. J Antibiot (Tokyo), 1986 Jan, 39(1), 136 - 40 Effects of cations, polyamines and other aminoglycosides on gentamicin C2 . Binding to ribosomes from sensitive and resistant Escherichia coli strains; Moukaddem M et al.; Gentamicin C2 interacts cooperatively with ribosomes from a sensitive Escherichia coli strain in a multiphasic way with several classes of sites . It is shown that this binding is highly-dependent on Mg++ and natural endogenous polyamine concentrations . The differences observed between ribosomes from sensitive and resistant strains may be explained by the absence of specific cooperative gentamicin interactions with resistant ribosomes . The effects of other aminoglycoside antibiotics are discussed in terms of structure-activity relationships. CRC Crit Rev Biochem, 1986, 19(3), 191 - 245 Kinetics of protein-nucleic acid interactions: use of salt effects to probe mechanisms of interaction; Lohman TM; The kinetics of protein-nucleic acid interactions are discussed with particular emphasis on the effects of salt concentration and valence on the observed rate constants . A general review is given of the use of experimentally determined salt dependences of observed kinetic parameters as a tool to probe the mechanism of interaction . Quantitative analysis of these salt dependences, through the application of polyelectrolyte theory, can be used to distinguish reactions which occur in a single step from those reactions which involve distinct intermediates . For those rate constants which display a large salt dependence, in either the association or dissociation reaction, this is due to the high concentration of counterions (e.g., Na+) in the vicinity of the nucleic acid which are subsequently released (or bound in the case of dissociation) at some point before the rate limiting step of the reaction . A general discussion of other features which affect protein-nucleic acid kinetics, such as nucleic acid length and the ratio of nonspecific to specific DNA binding sites (in the case of sequence specific binding proteins), is also given . The available data on the nucleic acid binding kinetics of small ligands (ions, dyes, oligopeptides), nonspecific binding proteins (T4 gene 32 protein, fd gene 5 and Escherichia coli SSB), and sequence specific binding proteins (lac repressor, RNA polymerase, Eco RI restriction endonuclease) are discussed with emphasis on the interpretation of the experimentally determined salt dependences. Chem Biol Interact, 1986 Jan, 57(1), 41 - 53 Efficient breakage of DNA apurinic sites by the indoleamine related 9-amino-ellipticine; Malvy C et al.; The aromatic amine, 9-NH2-ellipticine, is a synthetic DNA intercalating derivative of the antitumor agent ellipticine, which breaks circular DNA containing apurinic sites . This breakage is inhibited when the apurinic (AP) sites are reduced . The concentration of 9-NH2-ellipticine required to get a significant effect (0.1 microM) is the lowest known among chemicals which induce the same breakage reaction . Comparison with the action of structurally related amines shows that the amino-indole structure is specific for AP sites . The ability of ellipticine derivatives to induce breakage in DNA containing apurinic sites is related to the nucleophile substituent in position 9 . Two ellipticine derivatives with known antitumor activity, BD 40 and 9-OH-ellipticine, were able to break purified DNA at apurinic sites. Arch Biochem Biophys, 1986 Jan, 244(1), 218 - 25 Spatial relationship between the intrinsic metal in the beta subunit and cysteine-132 in the sigma subunit of Escherichia coli RNA polymerase: a resonance energy transfer study; Chatterji D et al.; Fluorescence excited-state energy transfer measurements were carried out between the N-(1-pyrene)maleimide (PM)-labeled sigma subunit and Co in the beta subunit of Co-Zn RNA polymerase (RPase) . sigma subunit with or without PM labeling was cleaved with 2-nitro-5-thiocyanobenzoic acid, and the reaction products were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . One molecule of the fluorescent probe (PM) was found to be attached to the cysteine-132 residue of the sigma subunit . When excited at 340 nm, the fluorescence emission bands from 380 to 420 nm of PM-labeled sigma overlap with the charge transfer absorption band of Co-Zn RPase around 400 nm . Based on Forster's equation, the R0 values for the donor-acceptor pair were calculated to be 21.5 and 22 A in the absence and presence of template analog (dA-dT)60, respectively . Using these R0 values and the observed energy transfer efficiencies, the distance between the cysteine-132 of the sigma subunit and Co located at the initiation site of the beta subunit was calculated to be 22 A with or without the template present, indicating that no major conformational change of the enzyme was induced upon template binding . However, a small but significant change in the above distance was observed upon the addition of ATP to RPase in the presence (dA-dT)60 but not in the absence of (dA-dT)60 template . The biological implications of these observations are discussed. Arch Biochem Biophys, 1986 Jan, 244(1), 128 - 36 The 18O isotope effect in 13C nuclear magnetic resonance spectroscopy: mechanistic studies on asparaginase from Escherichia coli; Rohm KH et al.; The mechanism of the enzyme asparaginase (L-asparagine amidohydrolase, EC 3.5.1.1) from Escherichia coli was examined using 13C NMR spectroscopy . The pH-dependent oxygen exchange reactions between water and aspartic acid were followed by use of the 18O isotope-induced shift of the resonance positions of directly bonded 13C nuclei . Both L-1- and L-1,4-{13C}aspartic acid were used in experiments with previously 18O-labeled aspartic acid, or in experiments involving the use of 18O-labeled solvent water . Asparaginase catalyzes a relatively efficient exchange between the oxygens of water and those on one carboxyl group of aspartic acid . Exchange at C-4 occurs rapidly but, within experimental error, no exchange at C-1 could be detected . These and related experiments involving the position of 18O incorporation during hydrolysis of aspartic acid beta-methyl ester are all consistent with possible acyl-enzyme mechanisms involving C-4, but do not support a free aspartic acid anhydride mechanism. Antibiot Med Biotekhnol, 1986 Jan, 31(1), 40 - 3 {Comparative study of the chemotherapeutic effectiveness of mecillinam, ampicillin and their combination in coli bacillary pyelonephritis in rats}; Zinchenko TA et al.; The chemotherapeutic effect of mecillinam and ampicillin was studied comparatively on rats with hematogenic obturation colibacillary pyelonephritis . The antibiotics were administered intragastrically in a dose of 100 mg/kg for 7 days . The treatment was started 24 hours after infection . When the drugs were used in combination their doses were twice as lower . When used alone mecillinam and ampicillin had a significant effect which was practically the same . A marked increase in the therapeutic effect was observed with the use of the antibiotics in combination: sterilization of the tissues of the affected kidney and prevention of development of macroscopic lesions in it in all the animals. South Med J, 1986 Jan, 79(1), 41 - 6 Emphysematous pyelonephritis: diagnosis and treatment; Klein FA et al.; Emphysematous pyelonephritis is a rare life-threatening bacterial infection usually occurring in patients with diabetes mellitus and producing gas within the renal parenchyma and/or perirenal tissue . We add four cases to the 62 previously reported in the literature . The overall mortality is 38%, with only 29% survival for those treated medically compared to 71% for those given both medical and surgical treatment . Early diagnosis with the aid of renal ultrasonography and/or computerized axial tomography and aggressive early combined medical-surgical management are emphasized. Nephron, 1986, 42(2), 171 - 6 Progression of chronic pyelonephritis in the rat; Mackenzie R et al.; Thirteen of 20 female Wistar rats developed perihilar kidney scars 6 weeks after ascending infection with Escherichia coli 078 . After removal of the lesser scarred kidney from 6 of the animals, all animals were followed for 54 weeks . Proteinuria (greater than 18 mg/24 h) developed at 20 weeks in the uninephrectomised infected rats and at 34 weeks in the 2-kidney infected model . In 10 uninephrectomised controls significant proteinuria did not appear until 52 weeks . In 9 2-kidney controls proteinuria did not develop at all . The speed of onset and severity of proteinuria was related to the extent of the renal parenchymal loss . Pyelonephritic scars did not show macroscopic progression over the 54-week observation period, even though the original renal infection persisted . Uninephrectomised animals with infected scars developed a highly significant rise of creatinine/body weight (p less than 0.02) and of heart weight/body weight p less than 0.02) ratios compared with the non-infected controls . Their kidneys showed focal and segmental hyalinosis and sclerosis of the glomeruli adjacent to the scars . Immunofluorescent staining for serum proteins was negative, but mesangial deposits of Tamm-Horsfall protein were found in the glomeruli between the scars in animals with renal impairment . These findings establish that progressive renal impairment in rats with infected kidney scars is associated with the development of proteinuria and a glomerulopathy . The cause of the glomerulopathy is not clear, both glomerular hyperfiltration and deposition of Tamm-Horsfall protein in glomerular mesangial cells may be involved. Nephron, 1986, 42(2), 128 - 32 Endotoxin level of sterile injection solutions and substitution fluid for hemofiltration in Japan and Australia; Tominaga H et al.; The endotoxin contamination levels of sterile distilled water and saline solutions used for injection and hemofiltration in Japan and Australia were examined with a colorimetric limulus test using a chromogenic substrate . The endotoxin levels of injection solutions in most products from both countries measured against E . coli 0111:B4 endotoxin or USP reference standard endotoxin were less than 0.2 pg or 0.0006 endotoxin units (EU) X ml-1 . Only 2 solutions in 30 from both countries showed higher levels: 7.7 and 10.8 pg for E . coli 0111:B4 endotoxin X ml-1 (0.02 and 0.03 EU X ml-1) . Even these higher values were well below the level recommended by the draft guideline published by the United States Food and Drug Administration (FDA) . The endotoxin contamination level of a Japanese hemofiltration substitution fluid ranged from 8.2 to 9.2 pg E . coli 0111:B4 endotoxin X ml-1 (0.024-0.027 EU X ml-1). J Pediatr Gastroenterol Nutr, 1986 Jan, 5(1), 70 - 3 Ultrastructural and biochemical changes in human jejunal mucosa associated with enteropathogenic Escherichia coli (0111) infection; Taylor CJ et al.; A case of prolonged diarrhoea following Escherichia coli 0111 gastroenteritis is reported . Electron microscopy of the jejunal biopsy revealed effacement of the brush border and attachment of bacteria by pedestal formation . Specific activities of brush border enzymes showed marked depression of disaccharidases, zinc-resistant alpha-glucosidase, and alkaline phosphatase . In contrast, marker enzymes for basolateral membranes and endoplasmic reticulum were unaffected . The biochemical changes support the pathogenic mechanism suggested by ultrastructural studies previously reported. J Am Vet Med Assoc, 1986 Jan 1, 188(1), 57 - 9 High technology diagnostics: detection of enterotoxigenic Escherichia coli, using DNA probes; Maddox CW et al.; A relatively new technique termed colony blot hybridization appears to be a reliable replacement for the ligated porcine gut loop assay previously used to detect stable toxin-B producing Escherichia coli . The highly reproducible blot assay enables screening of large numbers of isolants for the presence of E coli stable-B toxin genes, thus avoiding variations in phenotypic expression and poor repeatability inherent in the in vivo assays . Availability of this diagnostic test can aid practitioners in identifying enterotoxigenic E coli related disease in swine and in determining the benefits of vaccination programs. Environ Mutagen, 1986, 8(1), 161 - 9 The reproductive effects assessment group's review of the mutagenicity of vinylidene chloride; Jacobson-Kram D; A large number of studies indicate that vinylidene chloride is mutagenic to bacteria and that this activity is largely dependent on microsomal activation . Vinylidene chloride gave positive results for gene reversion and conversion in yeast that was also dependent on metabolic activation, and was positive in tradescantia . In mammalian systems, vinylidene chloride failed to induce gene mutations in V79 cells at two separate loci, failed to induce chromosomal aberrations in mouse bone marrow in vivo, and failed to induce dominant lethals in either mice or rats . Vinylidene chloride was found to alkylate DNA of mice exposed through inhalation and may have caused unscheduled DNA synthesis in kidneys of similarly exposed mice . The studies on the mutagenicity of vinylidene chloride are evaluated in this review. Circ Shock, 1986, 18(1), 31 - 42 Regional blood flow and metabolism in canine endotoxin shock before, during, and after infusion of glucose-insulin-potassium (GIK); Bronsveld W et al.; Glucose-insulin-potassium infused (GIK) during endotoxin shock causes increased cardiac output (CO) accompanied by decreased systemic vascular resistance . We have studied the effects of GIK on the distribution of cardiac output with radioactive microspheres to see if this decrease in resistance is equally distributed over all organs . GIK resulted in increased CO and increased flow to heart, splanchnic bed, kidneys, adrenals, and skeletal muscle, but fractional flow to these organs did not change . Thirty minutes after the GIK infusion, CO and organ flow had fallen again and differences between the endotoxin and control groups were no longer significant . GIK thus increases CO during endotoxin shock but does not influence its distribution . Systemic oxygen transport increased after GIK, but oxygen extraction decreased . Myocardial and splanchnic oxygen consumption did not change significantly . Oxygen extraction also diminished in these areas after GIK . GIK did not influence serum lactate: In both groups lactate increased significantly. Circ Shock, 1986, 18(1), 21 - 9 Lipolytic patterns in isolated adipocytes of continuously endotoxemic rats; Spitzer JA et al.; Lipolytic patterns were studied in adipocytes isolated from rats after 6 and 30 hr of continuous Escherichia coli endotoxin (ET) or saline infusion via a subcutaneously implanted osmotic pump . By 6 hr, ET cells responded to norepinephrine (NE) stimulation with significantly greater increase above basal rates of glycerol and free fatty acid (FFA) release than did control adipocytes . By 30 hr of continuous infusion, basal glycerol release was enhanced; the in vitro lipolytic response evoked by NE was significantly reduced in ET cells compared to rates on the previous day, and NE-stimulated lipolysis in ET cells was significantly below that of controls . At the same time, the in vitro antilipolytic effect of insulin was attenuated . We conclude that 1) an initial metabolic response can be observed within a few hours of a continuous, low dose ET infusion, 2) the biphasic nature of the sequential changes in lipolysis is likely to reflect alterations in the hormonal environment in vivo, and 3) these features are consonant with some aspects of the metabolic profile of septic patients. Circ Shock, 1986, 18(1), 11 - 9 Ultrastructural studies of experimental endotoxin shock in the liver and spleen: therapeutic effects of low-dose heparin on reticuloendothelial disturbances; Onda M et al.; To determine the effects of low-dose heparin on the reticuloendothelial system in shock, rats were subjected to intraperitoneal administration of E coli endotoxin . Alterations such as degeneration and necrosis of the Kupffer cells and sinusoidal endothelial cells were observed in the livers of the non-heparin-treated animals 4 hours after the administration of the endotoxin . These changes progressed with time . Kupffer cells in the heparin-treated rats appeared undisturbed up to 8-12 hours post endotoxin challenge . Phagocytosis of cell remained slight in the heparin-treated rats . The mortality rate of the heparin-treated group was 10% with LD50 endotoxin (7.6 mg/kg) . The mortality rate of the untreated control group 24 hours after administration was 50% . However, when endotoxin was administered in a dosage of 30 mg/kg (MLD), the mortality rate of the heparin-treated rats was 60%--10% greater than that of controls (P less than 0.05). Arch Surg, 1986 Jan, 121(1), 65 - 70 In vitro reversal of cardiac deterioration in septic shock with tetraethylammonium chloride; DeMeules JE; Overwhelming sepsis associated with cardiac failure continues to be a major clinical problem . This is commonly associated with a failure to respond to conventional pharmacologic therapy . This study was undertaken to see if manipulations of the electrophysiologic defects previously described by treatment with tetraethylammonium chloride (TEA) would be advantageous . Septic shock was induced in rabbits by a lethal dose of Escherichia coli . Peak tension and velocities of contraction and relaxation were measured in papillary muscle with and without 5mM TEA . Exposure to this compound improved peak tension and velocities of contraction and relaxation to normal values . The action of TEA is not specific to septic tissue as values in normal muscles are similarly improved . Tetraethylammonium chloride or other drugs that decrease outward potassium current and prolong the action potential duration may be helpful in treating cardiac dysfunction that accompanies sepsis. Am Rev Respir Dis, 1986 Jan, 133(1), 62 - 7 Reduction of total hemolytic complement activity with Naja haje cobra venom factor does not prevent endotoxin-induced lung injury in sheep; Flick MR et al.; We studied the effects of reducing total hemolytic complement activity with Naja haje cobra venom factor on the lung injury caused by intravenously infused endotoxin in 5 unanesthetized sheep with lung lymph fistulas . In normal sheep, infusions of lipopolysaccharide W from Escherichia coli (1.0 micrograms/kg) intravenously over 30 min caused increases in protein-rich lung lymph flow as well as the appearance in plasma and lung lymph of complement (C5)-derived chemotactic activity for polymorphonuclear leukocytes . Reduction of total hemolytic complement activity by treatment with Naja haje cobra venom factor (12 to 17 U/kg intraperitoneally) did not prevent the lung injury caused by endotoxin and also did not prevent the appearance in plasma and lung lymph of chemotactic activity . We conclude that although complement appears to be activated following intravenously infused endotoxin in sheep, a completely intact complement system is not necessary for endotoxin-induced lung injury. Proc Natl Acad Sci U S A, 1986 Jan, 83(2), 483 - 7 Design of a photoreactive analogue of the Escherichia coli heat-stable enterotoxin STIb: use in identifying its receptor on rat brush border membranes; Gariepy J et al.; The Escherichia coli heat-stable enterotoxin, STIb was prepared by solid-phase peptide synthesis and purified to homogeneity by high-pressure liquid chromatography . This analogue was iodinated and shown to bind specifically to rat intestinal membranes . The radiolabeled peptide was derivatized at the amino terminus with the photoreactive heterobifunctional crosslinking agent N-hydroxysuccinimidyl p-benzoylbenzoate . This photoreactive probe also exhibited binding specificity . It was mixed with rat intestinal brush border membranes and photolyzed in the presence or absence of excess unlabeled STIb . Polyacrylamide gel electrophoresis performed in the presence of sodium dodecyl sulfate and 2-mercaptoethanol indicated that the peptide probe was crosslinked specifically to two molecular species of 57 and 75 kDa . One or both of these molecules appear to constitute the enterotoxin receptor or to be in close proximity to it. Proc Natl Acad Sci U S A, 1986 Jan, 83(2), 303 - 7 Evidence for a contact between glutamine-18 of lac repressor and base pair 7 of lac operator; Ebright RH; Glutamine-18 of the lac repressor (lacR) has been substituted by glycine, by serine, and by leucine . The specificities of wild-type lacR and of the three substituted lacR variants have been analyzed with respect to base pairs 5, 6, 7, 8, 9, and 10 of the lac operator (lacO) . The data indicate that {Gly18}lacR, {Ser18}lacR, and {Leu18}lacR lose the ability to distinguish between the O+ base pair G . C and the Oc base pairs T . A and A . T at position 7 of lacO (KdOc/KdO+ approximately equal to 1) . In contrast, the three substituted variants retain the ability to discriminate O+ from Oc at each other position, by factors of 9 to 37 . Therefore, I propose that glutamine-18 contacts base pair 7 of lacO . These data suggest that the interaction between the helix-turn-helix motif and DNA may be very similar or identical in lacR and the catabolite gene activator protein. Proc Natl Acad Sci U S A, 1986 Jan, 83(2), 275 - 9 DNA hybridization electron microscopy: ribosomal RNA nucleotides 1392-1407 are exposed in the cleft of the small subunit; Oakes MI et al.; The ribosomal sequence corresponding to Escherichia coli 16S rRNA nucleotides 1392-1407 (the "1400 region") is phylogenetically conserved and is in a functionally important region of the subunit . Using the technique of DNA hybridization electron microscopy, we have mapped this sequence on the surface of the small ribosomal subunit . In this procedure a synthetic oligodeoxynucleotide probe, complementary to a specific rRNA sequence and carrying an attached marker molecule, is hybridized to ribosomal subunits in order to determine the dimensional site of attachment . In the E . coli ribosome, the 1400 region is located at the level of the neck, near the cleft and most likely on the head of the small subunit . The related sequence in yeast 18S rRNA, nucleotides 1618-1633, is located in the topological equivalent of the E . coli site . The location of this region, which has been crosslinked to the anticodon of a peptidyl-site-bound tRNA, indicates that this part of the cleft of the small subunit has a similar three-dimensional organization in phylogenetically diverse organisms and suggests that it is the site of the codon-anticodon interaction. Proc Natl Acad Sci U S A, 1986 Jan, 83(2), 231 - 5 Mechanism of the idling-turnover reaction of the large (Klenow) fragment of Escherichia coli DNA polymerase I; Mizrahi V et al.; The mechanism of the idling-turnover reaction catalyzed by the large (Klenow) fragment of Escherichia coli DNA polymerase I has been investigated . The reaction cycle involved is one of excision/incorporation, in which the 3' deoxynucleotide residue of the primer DNA strand is partitioned into its 5'-mono- and 5'-triphosphate derivatives, respectively . Mechanistic studies suggest the 5'-monophosphate product is formed in the first step by simple 3'----5' exonucleolytic cleavage . Rapid polymerization follows with the concomitant release of inorganic pyrophosphate . In the second step, the 5'-triphosphate product is generated by a pyrophosphorolysis reaction, which, despite the low concentration of pyrophosphate that has accumulated, occurs at a rate that is comparable with that of the parallel 3'----5' hydrolysis reaction. Mutat Res, 1986 Jan, 173(1), 13 - 8 Studies on mutagenesis and repair induced by platinum analogs; Fram RJ et al.; Mutagenesis and cytotoxicity were studied in Escherichia coli by iproplatin and carboplatin, two analogs of cisplatin (CDDP) currently undergoing clinical trial . As with CDDP, mutagenesis by these agents was mediated by the umuDC gene product . In contrast to CDDP, however, mismatch repair did not substantially contribute to survival of cells after exposure to these agents since dam-3 E . coli were not more sensitive than wild type E . coli . UvrA- E . coli, however were more sensitive to these analogs demonstrating that as with CDDP, uvr endonuclease-mediated excision contributes to the repair of DNA damage induced by platinum compounds. Mutat Res, 1986 Jan, 165(1), 39 - 44 Postreplicational formation and repair of DNA double-strand breaks in UV-irradiated Escherichia coli uvrB cells; Wang TC et al.; The number of DNA double-strand breaks formed in UV-irradiated uvrB recF recB cells correlates with the number of unrepaired DNA daughter-strand gaps, and is dependent on DNA synthesis after UV-irradiation . These results are consistent with the model that the DNA double-strand breaks that are produced in UV-irradiated excision-deficient cells occur as the result of breaks in the parental DNA opposite unrepaired DNA daughter-strand gaps . By employing a temperature-sensitive recA200 mutation, we have devised an improved assay for studying the formation and repair of these DNA double-strand breaks . Possible mechanisms for the postreplication repair of DNA double-strand breaks are discussed. Mutat Res, 1986 Jan-Feb, 159(1-2), 41 - 6 The molecular basis of the origin of complete and mosaic mutants; Dianov GL et al.; To study the molecular basis of the origin of complete and mosaic mutants, pBR322 plasmids with damage to one or both DNA strands were constructed by limited chemical modification of plasmid DNA . Damage to one strand of DNA resulted in the induction of predominantly mosaic mutants . Data were obtained indicating that complete mutations arise as a result of damage to both strands in the region of the mutated gene. Mutat Res, 1986 Jan-Feb, 159(1-2), 31 - 9 Inducible error-prone repair in yeast . Suppression by heat shock; Mitchel RE et al.; The production of reversion mutations in wild-type, diploid Saccharomyces cerevisiae by the alkylating agents N-methyl-N'-nitro- N-nitrosoguanidine (MNNG) and methylnitrosourea (MNU) was suppressed in cells previously treated with a heat shock, or the protein synthesis inhibitor, cycloheximide . The same cells previously treated with a heat shock, or the protein synthesis inhibitor, cycloheximide . The same treatment after mutagen exposure did not lower the induced mutation frequency . In split-dose experiments, a first MNNG exposure prevented subsequent heat (or cycloheximide) treatment from blocking mutation by a second, later mutagen exposure . These data suggest that, in yeast, MNNG or MNU induces an error-prone DNA-repair system, and that this induction is blocked by protein-synthesis inhibitors . The specificity of this system for different types of DNA damage was investigated using a variety of other mutagenic agents . A prior heat shock did not suppress mutation produced by exposure to ethyl methanesulfonate, ethylnitrosourea, 8-methoxypsoralen + UVA, or gamma-radiation . Partial suppression was observed in cells exposed to methyl methanesulfonate or to 254-nm ultraviolet light . These results indicate that, unlike the SOS system of E . coli, this inducible error-prone process of yeast is responsive to only certain mutagens . Heat shock suppression of mutation produced by MNNG exposure was also demonstrated in wild-type haploid cells, as well as haploid strains mutant in representative genes of the RAD52 epistasis group (rad52, rad53, rad54), the RAD3 epistasis group (rad1, rad2, rad3) and the RAD6 epistasis group (rad9, rad18) . The rad6 mutant itself was immutable with MNNG and therefore untestable by these techniques . These data indicate that this error-prone repair system is not absolutely dependent on the integrity of the RAD52 (recombination) or the RAD3 (excision) systems, or on at least some parts of the RAD6 system. Mutat Res, 1986 Jan-Feb, 159(1-2), 13 - 21 RecBC enzyme activity is required for far-UV induced respiration shutoff in Escherichia coli K12; Swenson PA et al.; Shutoff of respiration is one of a number of recA+ lexA+ dependent (SOS) responses caused by far ultraviolet (245 nm) radiation (UV) damage of DNA in Escherichia coli cells . Thus far no rec/lex response has been shown to require the recB recC gene product, the RecBC enzyme . We report in this paper that UV-induced respiration shutoff did not occur in either of these radiation-sensitive derivatives of K12 strain AB1157 nor in the recB recC double mutant . The sbcB gene product is exonuclease I and it has been reported that the triple mutant strain recB recC sbcB has near normal recombination efficiency and resistance to UV . The sbcB strain shut off its respiration after UV but the triple mutant did not show UV-induced respiration shutoff; the shutoff and death responses were uncoupled . We concluded that respiration shutoff requires RecBC enzyme activity . The RecBC enzyme has ATP-dependent double-strand exonuclease activity, helicase activity and several other activities . We tested a recBC+ (double dagger) mutant strain (recC 1010) that had normal recombination efficiency and resistance to UV but which possessed no ATP-dependent double-strand exonuclease activity . This strain did not shut off its respiration . The presence or absence of other RecBC enzyme activities in this mutant is not known . These results support the hypothesis that ATP-dependent double-strand exonuclease activity is necessary for UV-induced respiration shutoff. J Virol, 1986 Jan, 57(1), 101 - 9 Hepatitis B virus polypeptide X: expression in Escherichia coli and identification of specific antibodies in sera from hepatitis B virus-infected humans; Meyers ML et al.; Sequence analysis of the hepatitis B virus (HBV) genome revealed the presence of an open reading frame (ORF X) which has the potential to encode a 154-amino acid polypeptide . A fusion protein containing 145 of the amino acids encoded by ORF X and 8 amino acids of beta-galactosidase was expressed and characterized in bacterial extracts . Immunoprecipitations with the ORF X fusion protein as a radioactively labeled antigen were performed to screen sera of humans infected with HBV for the presence of antibodies against ORF X-encoded determinants (anti-X) . Such antibodies were identified in 9 samples from a set of 26 sera characterized as positive for HBV surface antigen but were not found in 16 normal human sera . The data reported here demonstrate that sera from some patients with markers of HBV infection contain antibodies directed against the polypeptide encoded by ORF X . As such, these findings represent evidence that ORF X constitutes a gene, or a portion of a gene, which is expressed during HBV infection . Although there does not appear to be a direct relationship between anti-X and any individual markers of HBV infection, our data suggest that anti-X is more prevalent in HBV-positive sera containing antibodies to HBe3 antigen (anti-HBe3). J Infect Dis, 1986 Jan, 153(1), 98 - 108 Relation of structure to function for the U . S . reference standard endotoxin after exposure to 60Co radiation; Csako G et al.; The structure and function of the highly purified U.S . reference standard endotoxin (RSE) were studied after exposure to ionizing radiation from a 60Co source . With increasing doses of radiation, the trilaminar ribbon-like structure of untreated endotoxin exhibited focal swelling, after which only spherical particles were seen by electron microscopy . These morphological changes were paralleled by the respective loss of O-side chain repeating units and pieces of the R-core from the lipopolysaccharide molecules, as demonstrated by electrophoresis . The biologic function of the irradiated endotoxin was assessed with a variety of tests . At higher doses of radiation, a direct relation was observed between the degradation of the molecular and supramolecular structure and the loss of biologic function . At lower doses of radiation, however, there was variability among the functional assays in their rate of change with progressive irradiation of the RSE . The results suggest that the carbohydrate moiety plays an important role both in determining the supramolecular structure and in modulating certain biologic activities of bacterial endotoxins. J Bacteriol, 1986 Jan, 165(1), 94 - 100 Highly mobile DNA segment of IncI alpha plasmid R64: a clustered inversion region; Komano T et al.; When R64 DNA was digested with EcoRI, two DNA fragments not equimolar to the plasmid DNA were produced . A DNA region including these fragments was cloned (pKK009), and the pKK009 DNA sample was found to be a mixture of six or more DNA species with EcoRI, PstI, and AvaI cleavage sites at different positions, suggesting a complex rearrangement of DNA . When a part of the pKK009 DNA was removed by HindIII digestion, 33 different types of plasmids (pKK010-series plasmids) were obtained out of 58 clones tested, but no DNA rearrangement could be observed . On the basis of a comparison of the detailed restriction maps of these pKK010-series plasmids, we propose a model in which four DNA segments invert independently or in groups within the 1.95-kilobase region of R64, so that the arrangements of these four segments change randomly . The fixed pKK010-series plasmid DNA was again rearranged in the presence of R64, indicating that trans-acting gene function may be present to mediate the DNA rearrangement . The gene (tentatively designated as rci) was located on a 4.5-kilobase E9' fragment of R64. J Bacteriol, 1986 Jan, 165(1), 82 - 7 Plasmid segregation into minicells is associated with membrane attachment and independent of plasmid replication; Rashtchian A et al.; The role of plasmid replication in the segregation of plasmids into Escherichia coli minicells was investigated with temperature-sensitive replication mutants derived from E . coli plasmids ColE1 and pSC101 . For as long as six generations of growth, at permissive or nonpermissive temperatures (when greater than 80% of plasmid replication was inhibited), the same amount of previously 3H-labeled plasmid DNA segregated into minicells . Density gradient separations of wild-type and temperature-sensitive plasmid DNA from both replicons segregated into the minicells showed that about 20 to 25% was stably associated with the minicell membrane at both temperatures . Electron microscopy showed this DNA to consist of circular plasmid molecules attached to the minicell membrane . These combined findings suggest that segregation of plasmids into minicells and their association with the minicell membrane are interrelated and independent of plasmid replication. J Bacteriol, 1986 Jan, 165(1), 34 - 40 Mutations in tar suppress defects in maltose chemotaxis caused by specific malE mutations; Manson MD et al.; Maltose-binding protein (MBP), which is encoded by the malE gene, is the maltose chemoreceptor of Escherichia coli, as well as an essential component of the maltose uptake system . Maltose-loaded MBP is thought to initiate a chemotactic response by binding to the tar gene product, the signal transducer Tar, which is also the aspartate chemoreceptor . To study the interaction of MBP with Tar, we selected 14 malE mutants which had specific defects in maltose taxis . Three of these mutants were fully active in maltose transport and produced MBP in normal amounts . The isoelectric points of the MBPs from these three mutants were identical to (malE461 and malE469) or only 0.1 pH unit more basic than (malE454) the isoelectric point of the wild-type protein (pH 5.0) . Six of the mutations, including malE454, malE461, and malE469, were mapped in detail; they were located in two regions within malE . We also isolated second-site suppressor mutations in the tar gene that restored maltose taxis in combination with the closely linked malE454 and malE461 mutations but not with the malE469 mutation, which maps in a different part of the gene . This allele-specific suppression confirmed that MBP and Tar interact directly. J Bacteriol, 1986 Jan, 165(1), 198 - 203 Posttranscriptional autoregulation of Escherichia coli threonyl tRNA synthetase expression in vivo; Butler JS et al.; Five mutations in thrS, the gene for threonyl-tRNA synthetase, have been characterized, and the sites of the mutations have been localized to different regions of the thrS gene by recombination with M13 phage carrying portions of the thrS gene . Quantitative immunoblotting shows that some of these mutations cause the overproduction of structurally altered threonyl-tRNA synthetase in vivo . The amounts of in vivo thrS mRNA as measured by quantitative hybridization are, however, the same as wild-type levels for each mutant . These results demonstrate that the expression of threonyl-tRNA synthetase is autoregulated at the posttranscriptional level in vivo. J Bacteriol, 1986 Jan, 165(1), 161 - 6 Nucleotide sequence corresponding to five chemotaxis genes in Escherichia coli; Mutoh N et al.; The nucleotide sequence of DNA which contains five chemotaxis-related genes of Escherichia coli, cheW, cheR, cheB, cheY, and cheZ, and part of the cheA gene was determined . Molecular weights of the polypeptides encoded by these genes were calculated from translated amino acid sequences, and they were 18,100 for cheW, 32,700 for cheR, 37,500 for cheB, 14,100 for cheY, and 24,000 for cheZ . Nucleotide sequences which could act as ribosome-binding sites were found in the upstream region of each gene . After the termination codon of the cheW gene, a typical rho-independent transcription termination signal was observed . There are no other open reading frames long enough to encode polypeptides in this region except those which code for the two previously reported genes tar and tap. Fed Proc, 1986 Jan, 45(1), 19 - 24 Endotoxin-mediated pulmonary endothelial cell injury; Meyrick BO; Infusion of endotoxin into sheep results in physiological and structural damage to the pulmonary endothelium . It is uncertain whether complement activation and granulocyte sequestration in the pulmonary microcirculation and the ensuing granulocyte migration into the interstitium seen with endotoxemia contribute to the endothelial damage . We have shown that infusion of complement-activated plasma into sheep, although causing the same degree of granulocyte sequestration in the lungs, results in only modest and transient endothelial damage . In addition, migration or chemotaxis of granulocytes across the endothelial layer of intimal explants is not accompanied by either structural evidence of endothelial damage or a detectable increase in vascular permeability . Such studies indicate that neither complement/granulocyte activation nor granulocyte migration across a vessel wall is entirely responsible for the severe endothelial damage seen with endotoxin . In vitro studies of bovine pulmonary endothelial monolayers indicate that endotoxin can cause direct damage to the endothelium; the damage is dose-dependent and more severe in the presence of serum . Structural studies show endothelial cell retraction, pyknosis, and sloughing . Prostacyclin production and lactic dehydrogenase release are increased, as are permeability to small solutes and hydraulic conductance across the endothelium . It seems that endotoxin can cause a direct injury to pulmonary endothelium but complement and granulocyte activation may enhance the damage. FEBS Lett, 1986 Jan 1, 194(1), 12 - 5 The roles of HPr and FPr in the utilization of fructose by Escherichia coli; Kornberg H; A mutant impaired in FPr activity was isolated . The altered gene (fpr), which was located near min . 44 on the E . coli genome, was transferred by phage-mediated transduction to appropriate recipients that lack HPr (ptsH), or Enzyme IIman (ptsM), or neither . The rates of growth on fructose of such transductants indicate that phosphate from PEP is transferred predominantly via FPr to fructose that enters the cells by Enzyme IIfru, but that HPr can play a role in transferring phosphate to fructose taken up via Enzyme IIman. Free Radic Res Commun, 1986, 2(1-2), 27 - 42 Anti-inflammatory activity of superoxide dismutases: studies on adjuvant induced polyarthritis in rats; Jadot G et al.; The anti-arthritic activities of various superoxide dismutases and of liposomal bovine Cu-SOD have been compared in the adjuvant induced Lewis Inbred Rat model . Various approaches, including plethysmometric measurements, red cell sedimentation rates, while cell counts, levels of IgA and IgG