J Clin Microbiol, 1990 May, 28(5), 1071 - 4 Cellular fatty acid compositions of an unidentified organism and a bacterium associated with cat scratch disease; Moss CW et al.; The cellular fatty acid composition of a gram-negative bacterium associated with cat scratch disease was determined by capillary gas chromatography-mass spectrometry (GC-MS) . The fatty acid profile of this organism was distinct from those of other bacteria we have tested and was characterized by an unknown acid which was identified as 11-methyloctadec-12-enoic acid . The position of the branched methyl group in this acid was established by GC-MS of the reduced acid, and the location of the double bond was confirmed by GC-MS analysis of dimethyl disulfide derivatives . Another clinical isolate with no known relationship to cat scratch disease but with similar morphological and biochemical features had a similar fatty acid profile, including 11-methyloctadec-12-enoic acid. Kidney Int, 1990 May, 37(5), 1219 - 26 Direct effects of endotoxin on the function of the isolated perfused rat kidney; Cohen JJ et al.; When the endotoxin-lipopolysaccharide (LPS) derived from the cell wall of gram-negative bacteria is given to the rat in vivo, there are prompt, marked decreases in glomerular filtration rate (GFR), renal blood flow (RBF) and % Na+ reabsorption (%T-Na+) . However, it has not been determined whether the endotoxin itself has a direct effect on these renal functions . To test whether endotoxin has a direct renal effect, isolated rat kidneys (N = 8) were perfused for 160 minutes with a Krebs-Ringer-HCO3- solution containing substrate-free albumin (40 g/liter), glucose (5 mM) and L(+) lactate (7.5 mM) . After control observations (20 to 80 min) were made, purified LPS from E . coli was added (N = 4) to the perfusate to achieve {endotoxin} of 0.01 micrograms/ml (80 to 120 min) and 0.1 micrograms/ml (120 to 160 min) . Endotoxin had no effect on GFR, Na+ reabsorption or tissue K+ content when compared to timed-control perfusions (N = 4) . There was a small (approximately 10%) but significant decrease in mean perfusion flow rate (PFR) at the highest {endotoxin} when compared to the low {endotoxin}p but no change in GFR occurred . When the same LPS was given to four rats in vivo at a dose which achieved an {endotoxin} of approximately 0.08 micrograms/ml plasma, there were prompt decreases in GFR and %T-Na+ and an increase in body temperature when compared with timed-controls; there also was a large loss of K+ from the kidney tissue.(ABSTRACT TRUNCATED AT 250 WORDS) Ann Surg, 1990 May, 211(5), 543 - 9; discussion 549-51 The effects of sepsis and endotoxemia on gut glutamine metabolism; Souba WW et al.; The effects of sepsis on gut glutamine (GLN) metabolism were studied to gain further insight into the regulation of the altered glutamine metabolism that characterizes critical illnesses . Studies were done in laboratory rats and in hospitalized patients . The human studies were done in seven healthy surgical patients (controls) and six septic patients who underwent laparotomy . Radial artery and portal vein samples were obtained during operation and were analyzed for GLN and oxygen content . Despite no reduction in arterial glutamine concentration in the septic patients, gut glutamine extraction was diminished by 75% (12.0% +/- 1.6% in controls vs . 2.8% +/- 0.8% in septic patients, p less than 0.01) . Similarly gut oxygen extraction was diminished by nearly 50% in the septic patients (p less than 0.05) . To further investigate these abnormalities, endotoxin (10 mg/kg intraperitoneally) or saline (controls) was administered to adult rats 12 hours before cannulation of the carotid artery and portal vein . The arterial GLN concentration was increased by 13% in the endotoxin-treated animals (p less than 0.05) but gut glutamine uptake was diminished by 46% (526 +/- 82 nmol/100 g BW/minute in controls vs . 282 +/- 45 in endotoxin, p less than 0.01) . Simultaneously gut glutaminase activity was diminished by 30% (p less than 0.01) and intestinal glutamate release fell by two thirds . Blood cultures were negative in control animals (0 of 20), but were positive in 25% of endotoxemic animals (6 of 24) for gram-negative rods (p = 0.019) . Sepsis and endotoxemia impair gut glutamine metabolism . This impairment may be etiologic in the breakdown of the gut mucosal barrier and in the development of bacterial translocation. J Indian Med Assoc, 1990 May, 88(5), 131 - 3 Pulmonary complications of septicaemia in women; Malhotra M et al.; Twenty-five female patients of septicaemia aged 15-50 years comprised this study . Twenty-three patients (92%) were in the reproductive age group (15-35 years) . Sixteen patients (64%) had some relation to parity, suggesting an increased risk of septicaemia in women with puerperal sepsis . The commonest primary source of infection was the female genital tract (48%) . Gram-negative septicaemia was the commonest . The pleuropulmonary complications observed were pneumonia (10 cases), lung abscess (7 cases), empyema (3 cases), septic pulmonary embolisation (4 cases) and adult respiratory distress syndrome (4 cases) . Seven cases of the total 25 died . Adult respiratory distress syndrome with septicaemia was the commonest cause of mortality in these patients. Pathol Biol (Paris), 1990 May, 38(5), 436 - 40 {Interaction of bactericidal serum effect and antibiotics in subminimal inhibitory concentrations on E . coli strains}; Gillissen G et al.; Imipenem (IMI) forming round cells in Gram negative rods reduces in subinhibitory concentrations (subMIC) the seroresistance of E . coli . This effect is distinctly more pronounced in a moderately seroresistant strain of E . coli than in a high seroresistant one . Conversely, human serum (HS) increases the sensitivity of E . coli strains to IMI dependent on their original seroresistance . In contrast, ampicillin (AMP), a filament inducer in E . coli, reduces equally seroresistance but only to a minimal degree and that only in a moderately seroresistant strain; the high seroresistant strain was nonreactive in this respect . It was concluded, that a synergism of antibiotics and bactericidal serum effect is predominantly produced with round cell forming antibiotics, whereas filament forming ones show only minimal effects . Moreover, the original seroresistance of strains in apparently important for the degree tho which these phenomena are expressed. Appl Environ Microbiol, 1990 May, 56(5), 1271 - 8 Comparison of the limulus amebocyte lysate test and gas chromatography-mass spectrometry for measuring lipopolysaccharides (endotoxins) in airborne dust from poultry-processing industries; Sonesson A et al.; The lipopolysaccharide (endotoxin) content in airborne dust samples from three different poultry slaughterhouses was determined with both the chromogenic Limulus amebocyte lysate assay and gas chromatography-mass spectrometry analysis of lipopolysaccharide-derived 3-hydroxy fatty acids . Gram-negative cell walls were also measured by using two-dimensional gas chromatography/electron-capture analysis of diaminopimelic acid originating from the peptidoglycan . The correlation between the results of the Limulus assay and those of gas chromatography-mass spectrometry for determination of the lipopolysaccharide content in the dust samples was poor, whereas a good correlation was obtained between lipopolysaccharide and diaminopimelic acid concentrations with the gas chromatographic methods . The results suggest that it is predominantly cell-wall-dissociated lipopolysaccharides that are measured with the Limulus assay, whereas the gas chromatographic methods allow determination of total concentrations of lipopolysaccharide, including Limulus-inactive lipopolysaccharide, gram-negative cells, and cellular debris. Am Rev Respir Dis, 1990 May, 141(5 Pt 1), 1296 - 306 Effects of D-galactosamine-induced acute liver injury on mortality and pulmonary responses to Escherichia coli lipopolysaccharide . Modulation by arachidonic acid metabolites; Matuschak GM et al.; Multiple extrapulmonary organ system failures increase mortality, permeability edema, and alveolar inflammation during gram-negative sepsis because of abnormal regulation of host inflammatory responses . We tested the hypothesis that acute hepatocytic injury induced by the selective hepatotoxin, D-galactosamine (GalN), augments mortality and amplifies pulmonary microvascular permeability to albumin and neutrophilic influx after administering Escherichia coli lipopolysaccharide (LPS) 24 h later by impairing the metabolism of endogenously synthesized products of arachidonic acid . We determined the lung extravascular leak of 125I-human serum albumin measured at multiple time points after LPS and enumerated polymorphonuclear leukocytes (PMNs) in bronchoalveolar lavage fluid (BALF) . Because the liver is important in prostaglandin (PG) and leukotriene (LT) metabolism, we measured plasma concentrations of 6-keto-PGF1 alpha and thromboxane B2 (TxB2) in addition to paired plasma BALF concentrations of LTB4 and BALF LTC4 60 min and 24 h after LPS . We further assessed the protective effects of a single 20-mg/kg injection given intraperitoneally (i.p.) of the LTA4 synthetase inhibitor, diethylcarbamazine (DEC) . After 400 mg/kg GalN, LPS at 2.5 or 1.25 mg/kg i.p . increased mortality (p less than 0.001), albumin leak 60 and 90 min after LPS (p less than 0.05), plasma 6-keto-PGF1 alpha, TxB2, and LTB4 levels and BALF LTC4 within 60 min (p less than 0.05) . LTB4 and LTC4 levels in BALF 24 h later were similarly increased (p less than 0.05) as were bronchoalveolar PMNs (p less than 0.001) . DEC improved mortality and albumin leak (p less than 0.001), reduced lung influx of PMNs and peripheral leukocytosis (p less than 0.05), attenuated plasma LTB4 and BALF LTC4 levels 60 min after LPS (p less than 0.05), and decreased BALF LTB4 and LTC4 at 24 h (p less than 0.05), but was associated with higher plasma 6-keto-PGF1 alpha and TxB2 values at 60 min . Changes in eicosanoid levels and modulation of responses by DEC in this model suggest that impaired metabolism of endogenously synthesized leukotriences by the damaged liver underlies these phenomena . We conclude that this mechanism may enhance septic lung injury during acute liver dysfunction. J Bacteriol, 1990 May, 172(5), 2303 - 12 Interdomain hybrid Tet proteins confer tetracycline resistance only when they are derived from closely related members of the tet gene family; Rubin RA et al.; Inner membrane Tet proteins encoded by tet genes in gram-negative bacteria mediate resistance to tetracycline (Tcr) by directing its export . Total sequences for class A, B, and C tet genes demonstrate that their products have a common ancestor, with Tet(A) and Tet(C) being more closely related (78% identical) than either is to Tet(B) (45% identical) . The N- and C-terminal halves of Tet(B) and Tet(C) appear to comprise separate domains, and trans-complementation observed between tetracycline sensitive mutants in either domain of Tet(B) suggests separate but interactive functions for these domains . In this present study, interdomain hybrid genes were constructed to express hybrid tet products whose N- and C-terminal halves were derived from different family members {Tet(A/C), Tet(B/C), and Tet(C/B)} . Tet(A/C) specified a level of Tcr comparable to wild-type Tet(C) and 60% that of Tet(A), indicating that domains from these closely related tet products can function in cis . Although neither Tet(B/C) nor Tet(C/B) hybrids conferred significant Tcr, cells producing both of these types of hybrid proteins expressed substantial Tcr, indicating that productive interactions can occur in trans between Tet(B/C) and Tet(C/B) . Taken together, these results suggest that highly specific interactions between the N- and C-terminal domains are necessary for Tcr and do not occur in individual hybrids derived from the more distant relatives, Tet(B) and Tet(C) . This requirement for specific interactions suggests that N- and C-terminal domains have coevolved in each member of the Tet family. Res Microbiol, 1990 May, 141(4), 425 - 35 Comparison of beta-galactosidase production by two inducible promoters in Myxococcus xanthus; Letouvet-Pawlak B et al.; The inducibility of two promoter systems, one heterologous and one homologous, has been assessed in the Gram-negative bacterium Myxococcus xanthus . The heterologous system involved the hybrid tac promoter and the presence of lacIq, the lac repressor from Escherichia coli . This system is inducible in its natural host with isopropyl-beta-D-thiogalactopyranoside (IPTG) . The homologous promoter system involves the light-inducible carQRS promoter, which is normally involved in the expression of the regulators of the light-inducible light-protective carotenoid synthesis regulon in M . xanthus . In each case, promoter activity and strength was assayed using the E . coli gene lacZ . In our constructs, which were present in a single copy in the M . xanthus chromosome, the carQRS promoter yielded at least a 47-fold increase in beta-galactosidase production upon light induction, whilst IPTG increased by 8-fold the amount of enzyme produced under the control of the ptac-lacIq system . Regulation by the latter was significantly higher than that obtained with the unmodified lacZ promoter. J Appl Physiol, 1990 May, 68(5), 1820 - 5 Effects of hypercarbia on arterial and alveolar oxygen tensions in a model of gram-negative pneumonia; Keenan RJ et al.; Inspired CO2 causing changes from hypo- to normocapnia has previously been shown to improve arterial O2 tension (PaO2) and to reduce alveolar-arterial O2 difference . The effect of further increases in inspired CO2 to hypercarbic levels has not been studied in inflammatory lung disease . Three days after induction of sublobar Pseudomonas pneumonia, Suffolk sheep were anesthetized and ventilated with a fixed-volume ventilator . After 2.5 h, CO2 was added to the inspired gas to raise arterial CO2 tension (PaCO2) to 60-65 Torr . Four hours later the CO2 was withdrawn and ventilation continued for an additional 2 h . Constant minute ventilation and inspired O2 fraction were maintained . Regional lung perfusion was measured by injection of radioactive microspheres . With the administration of CO2, PaO2 increased significantly from 65.5 to 77.5 Torr as did alveolar O2 tension (from 109.7 to 120.0 Torr) with no significant change in alveolar-arterial O2 difference . There were no significant changes in cardiac output, shunt fraction, O2 uptake, O2 delivery, respiratory quotient, or distribution of regional lung perfusion . We conclude that the increases in alveolar O2 tension and PaO2 with the added CO2 resulted from improved alveolar ventilation. J Immunol, 1990 May 1, 144(9), 3506 - 12 Deacylated lipopolysaccharide inhibits plasminogen activator inhibitor-1, prostacyclin, and prostaglandin E2 induction by lipopolysaccharide but not by tumor necrosis factor-alpha; Riedo FX et al.; Bacterial LPS and TNF induce vascular endothelial cells to express a variety of response molecules . LPS that is partially deacylated (dLPS) by a human neutrophil enzyme blocks the ability of LPS, but not TNF, to augment one of these responses, the expression of endothelial cell surface molecules that promote neutrophil adherence (J . Exp . Med . 1987; 165:1393-1402) . We show that dLPS can inhibit the ability of LPS, but not TNF, to elicit the expression of plasminogen activator inhibitor-1 (PAI-1), prostacyclin, and PGE2 by human umbilical vein endothelial cells . dLPS also prevented the accumulation of specific PAI-1 mRNA in response to LPS, but not to TNF . Neither the LPS- or TNF-induced expression of PAI-1 nor the dLPS inhibition of the LPS response was mediated by prostanoids . These results indicate that dLPS can specifically block a variety of endothelial cell responses to LPS and provide support for the hypotheses 1) that dLPS and LPS may interact with a common target molecule on or in endothelial cells, and 2) that dLPS, produced by enzymatic deacylation of LPS in vivo, could inhibit endothelial cell stimulation by LPS and thereby limit the host inflammatory response to invasive gram-negative bacteria. Am J Respir Cell Mol Biol, 1990 May, 2(5), 433 - 40 Type I pili mediate gram-negative bacterial adherence to intact tracheal epithelium; Dal Nogare AR; We have investigated the role of pili in mediating gram-negative bacterial adherence to an intact tracheal epithelium . Type 1 pili, but not P or Pseudomonas pili, markedly increased bacterial adherence . The adherence-promoting effect of Type 1 pili was due to the mannose-binding Type 1 pili adhesin, as both alpha-methyl mannoside and concanavalin A blocked adherence of Type 1 piliated bacteria . The Type 1 pili-binding site on tracheal epithelium appears to be a mannose-containing glycoprotein . Clearance of Type 1 piliated bacteria from the lung parenchyma was assessed by depositing the bacteria into a lobe; no difference in clearance rates between Type 1 and nonpiliated bacteria was present . Type 1 pili may enhance the ability of gram-negative bacteria to adhere to and colonize the lower respiratory tract. Microb Pathog, 1990 May, 8(5), 353 - 62 Lipooligosaccharide epitopes shared among gram-negative non-enteric mucosal pathogens; Campagnari AA et al.; The non-enteric Gram-negative human pathogens, B . catarrhalis, H . ducreyi, H . influenzae, N . gonorrhoeae and N . meningitidis, do not have repeating O-antigens as part of their principle surface glycolipid, the lipooligosaccharide (LOS) . Because they have similar LOS structures, we studied the conservation of LOS oligosaccharide epitopes among these organisms . Twenty-one monoclonal antibodies (mAbs) generated by immunizing mice with H . influenzae, N . gonorrhoeae and N . meningitidis were studied for cross reactivity . Five mAbs generated against non-typable H . influenzae were the only strain-specific antibodies . Ten mAbs reacted to LOS epitope(s) common to a genera or species, and six mAbs bound to epitope(s) on the LOS of strains from different genera . Some cross reactive mAbs bound to LOS bands of similar molecular weights, while others bound to bands of varying molecular weights . mAb 3F11, whose epitope mimics a human blood-group antigen, bound to a 4.8 kDa LOS band in N . gonorrhoeae and H . ducreyi, two pathogens that infect genital epithelium . mAb 3D9, whose epitope consists of 2-keto-3-deoxyoctulosonic acid (KDO), reacted with different LOS bands in N . gonorrhoeae, H . influenzae and some R mutants of S . minnesota . A 14 kb restriction fragment containing lipooligosaccharide synthesis genes responsible for the assembly of the 3D9 epitope in H . influenzae hybridized to all H . influenzae strains tested but did not hybridize to gonococcal and S . minnesota strains that expressed this epitope . These studies demonstrate that conserved LOS epitope(s) exist among different species and genera of non-enteric human pathogens and that different genetic mechanisms may have evolved in these pathogens to assemble some of these conserved epitopes. Lett Appl Microbiol, 1990 May, 10(5), 209 - 12 An improved method for rapid isolation of plasmid DNA from wild-type gram-negative bacteria for plasmid restriction profile analysis; Olsen JE; An improved method for the isolation of large and small plasmids from wild-type Gram-negative bacteria has been developed . The protocol combines the lysis and purification procedures of two popular plasmid isolation methods, and produces DNA sufficiently pure for restriction enzyme digestion in less than three hours. Pharm Weekbl Sci, 1990 Apr 27, 12(2), 46 - 50 Helicobacter pylori in gastroduodenal disease; Loffeld RJ; Helicobacter pylori is a recently described Gram-negative micro-organism, which plays an important role in the pathogenesis of peptic ulcer disease and gastritis . In this paper an outline is given of the history and the pathophysiology of Helicobacter pylori, as well as the therapeutic studies aimed at eradication of the bacterium in various clinical entities. J Immunol Methods, 1990 Apr 17, 128(2), 219 - 25 An enzyme-linked immunosorbent assay for the determination of human IgG subclass antibodies directed against Branhamella catarrhalis; Goldblatt D et al.; An ELISA procedure to determine the distribution of human IgG subclass antibodies directed against the gram-negative bacterium Branhamella catarrhalis has been developed using commercially available monoclonal anti-IgG subclass antibodies . Using whole bacteria as coating antigen the specificity of the assay was determined and showed minimal cross-reactivity with a range of other bacteria . Estimations of IgG1, IgG2, IgG3, IgG4 and total IgG antibodies directed against this antigen were performed . All normal adult sera tested had measurable antibody levels of specific IgG1, IgG2, IgG3 and total IgG . Specific IgG4 was undetectable in the majority of adult sera . These assays will be of value for investigation of both children and adults with suspected immunodeficiency and recurrent upper respiratory tract infection. J Biol Chem, 1990 Apr 15, 265(11), 6394 - 402 A mutant of Escherichia coli defective in the first step of endotoxin biosynthesis; Galloway SM et al.; Using localized mutagenesis of whole cells, we have isolated a temperature-sensitive UDP-N-acetylglucosamine acyltransferase mutant of Escherichia coli that loses all detectable acyltransferase activity and quickly dies after a shift from 30 to 42 degrees C . Acyltransferase activity and temperature resistance are restored by transforming the mutant with a hybrid plasmid containing the E . coli gene for UDP-GlcNAc acyltransferase (lpxA) . In addition, a new assay has been developed for quantitating the amount of lipid A (the active component of endotoxin) in E . coli and related Gram-negative strains . Cells are labeled with 32Pi and extracted with chloroform/methanol/water (1:2:0.8, v/v) to remove glycerophospholipids . The residue is then hydrolyzed with 0.2 M HCl to liberate the "monophosphoryl" lipid A degradation products (Qureshi, N., Cotter, R . J . and Takayama, K . (1986) J . Microbiol . Methods 5, 65-77), each of which bears a single phosphate residue at position 4' . The amount of lipid A is normalized to the total amount of labeled glycerophospholipid present in the cells . The steady state ratio of lipid A to glycerophospholipid in wild-type cells is approximately 0.12 . The lipid A content of the acyltransferase mutant is reduced 2-3-fold, and the rate of lipid A synthesis is reduced 10-fold when compared to wild-type after 60 min at 42 degrees C . These results provide physiological evidence that UDP-N-acetylglucosamine acyltransferase is the major committed step for lipid A biosynthesis in E . coli and that lipid A is an essential molecule. Zh Mikrobiol Epidemiol Immunobiol, 1990 Apr, (4), 64 - 70 {The use of blood plasma with elevated titers of antibodies to Re-chemotype glycolipid in peritonitis}; Apollonin AV et al.; The article analyzes the possibility of immunotherapy of septic complications in cases of peritonitis caused by gram-negative bacteria . A strictly inverse correlation between the severity of intoxication and the level of antibodies to glycolipid Re in blood serum has been established . About 5% of healthy nonimmunized donors have elevated Re-antibody titers (1:128 and higher) in their blood plasma . Screening of blood preparations from more than 1000 donors permitted the creation of the blood plasma bank used for the treatment of peritonitis patients . Immunotherapy of such patients has made it possible to decrease almost two-fold the death rate and to reduce the severity and duration of the intoxication syndrome in patients, as well as to improve the results of the treatment of peritonitis. Jpn J Antibiot, 1990 Apr, 43(4), 692 - 5 {Basic and clinical studies on aztreonam in obstetric and gynecologic infections during the perinatal period}; Makinoda S et al.; The basic and clinical efficacies of aztreonam (AZT) were evaluated in obstetric and gynecologic infections during the perinatal period . The results obtained are summarized below . 1 . Concentrations of AZT in umbilical venous blood and amniotic fluid showed a good placental transfer of AZT . The rate of placental transfer of AZT was very similar to rates for cephalosporins . 2 . Since the clinical efficacy rate of AZT in the treatment of Gram-negative bacterial infections was 80.0%, AZT appears to be a useful drug against Gram-negative bacterial infections. Jpn J Antibiot, 1990 Apr, 43(4), 686 - 91 {Pharmacokinetic and clinical studies on aztreonam in perinatal period}; Sengoku K et al.; Pharmacokinetic and clinical studies were carried out on aztreonam (AZT), a monobactam antibiotic with a high activity against Gram-negative bacteria . The results obtained are summarized as follows: 1 . Following 2 g bolus intravenous injection, transfers of AZT to umbilical cord serum and amniotic fluid were found to be satisfactory . AZT level in amniotic fluid was higher than 1 micrograms/ml at 40 minutes after administration and it was at 3.7 micrograms/ml in 23.5 hours . 2 . In the treatment of 9 patients with perinatal infections, clinical efficacies of AZT were judged excellent in 3 cases and good in 6 cases . 3 . No side effects and abnormal laboratory findings due to the drug were observed in any case . These results indicate that AZT may be a useful antibiotic for the treatment of perinatal infections. Noseleutike, 1990 Apr-Jun, 29(132), 113 - 22 {Determinative factors in nosocomial pneumonia in patients with mechanical ventilation in the respiratory care unit}; Apostolopoulou E et al.; We studied risk factors for nosocomial pneumonia in 192 intubated patients . Nosocomial pneumonia was diagnosed in 50 (26%) of the 192 patients . The patients receiving mechanical ventilation and the ventilators were colonised with Gram negative bacteria, mainly Actinetobacter and Pseudomonas . The duration of mechanical ventilation and the bacterail colonization of ventilators were associated with the development of pneumonia. J Biol Response Mod, 1990 Apr, 9(2), 178 - 84 Initial evaluation of a human immunoglobulin M monoclonal antibody (HA-1A) in humans; Khazaeli MB et al.; A human monoclonal antibody (HA-1A) directed against bacterial endotoxin was administered to 15 patients with incurable malignant disease . No adverse effects were noted following single intravenous infusions of 0.05 to 100 mg . Pharmacokinetics were evaluated in nine patients receiving 10 mg (n = 3), 25 mg (n = 3), and 100 mg (n = 3) . Seven of these patients had initial peak serum concentrations greater than 80% of predicted values with plasma disappearance curves fitting a one-compartment system and a plasma half-life of 31.5 h (range of 20.3-44.6 h) . The peak serum concentrations and area under the curve values were proportional to the dose of HA-1A administered . One patient had a hypercatabolic state with low levels of serum albumin and IgM . He achieved 65% of the predicted value for peak serum concentration of HA-1A with a plasma half-life of 12.3 h . A second patient had detectable serum HA-1A for only 15 min following infusion without an adequate technical or biologic explanation . We were unable to demonstrate antibody to HA-1A in sera from these nine patients either prior to therapy or during 28 days postinfusion using a "double-antigen" radiometric assay . This study suggests that HA-1A human monoclonal antibody administration is well tolerated by patients . Phase I trials will need to be carried out to characterize further the pharmacokinetics and toxicity of HA-1A in patients with gram-negative sepsis. J Clin Microbiol, 1990 Apr, 28(4), 689 - 93 Biochemical and chemical characterization of pink-pigmented oxidative bacteria; Wallace PL et al.; The biochemical and chemical characteristics were determined for 156 clinical isolates of pink-pigmented bacteria that are similar to but distinct from Methylobacterium extorquens (synonymous with Pseudomonas mesophilica) . These isolates were gram-negative, nonfermentative, usually nonvacuolated, coccoid rods; all grew at 35 degrees C and were catalase and urease positive; the majority grew on MacConkey agar and were variable for oxidase production and motility . On the basis of oxidation of xylose and mannitol and hydrolysis of esculin, these 156 strains were subdivided into four groups that were designated "pink coccoid" groups I, II, III, and IV . Groups I, II, and III are similar to an unnamed taxon described by Gilardi and Faur in 1984; only strains of group IV hydrolyze esculin . The cellular fatty acid compositions of strains of groups I, II, and III were essentially identical and differed from strains of group IV by the absence of 3-OH-C14:0 and the presence of C19:0 delta and 2-OH-C19:0 delta . The fatty acid composition of group IV strains was most similar to that of M . extorquens but differed by the presence of small amounts of two C17:1 acids, 3-OH-C16:0, and 2-OH-C18:1. Cornea, 1990 Apr, 9(2), 108 - 14 Infectious crystalline keratopathy in a neonatal infant; Burnette WC et al.; Bilateral white crystalline opacities of the central corneas were found at autopsy in the eyes of a 17-day-old girl with Turner's Syndrome . Corticosteroid therapy had not been used . Histopathologic and electron microscopic evaluation revealed clusters of gram-negative rods between relatively intact stromal lamellae . Inflammation was mild and restricted to the limbus and peripheral cornea . This case represents an unusual presentation of an infectious crystalline keratopathy. Can J Vet Res, 1990 Apr, 54 Suppl, S53 - 6 Actinobacillus pleuropneumoniae: molecular aspects of virulence and pulmonary injury; Bertram TA; Contributions made by several laboratories in the area of Actinobacillus pleuropneumoniae virulence and its relationship to pulmonary disease will be reviewed briefly . Lung injury and subsequent disease, after infection with A . pleuropneumoniae, can be related to various bacterial toxins and host factors . Similar to other gram-negative bacteria . A . pleuropneumoniae has cell wall lipopolysaccharides which have been incriminated in a wide variety of toxic and tissue damaging processes . Virulent isolates of A . pleuropneumoniae have been shown to have a thick capsule whereas some avirulent isolates have a thin and easily removed capsule . The capsule of A . pleuropneumoniae is a linear unbranched polysaccharide composed of repeating dissaccharide subunits that bestow antiphagocytic properties to the bacterium but are also immunogenic . In addition, A . pleuropneumoniae has several chemically defined exotoxins . These toxins have generally been shown to be proteinaceous molecules that are hemolytic, cytotoxic, or edemogenic . Some of these toxins are proteolytic and others have the putative activity of being lytic for secretory IgA . Several of these molecules are capable of inducing lesions that are similar to those observed in natural infections and disease . Endogenous host factors have also been implicated in the development of lung lesions after infection by A . pleuropneumoniae Coagulation and inflammatory pathways have been demonstrated to be pivotal in the early phases of lesion development . In addition, the immune status of the animal is clearly related to the severity and ultimate outcome of A . pleuropneumoniae infection . To adequately treat and prevent this disease, we must understand the distinguishable interactions that occur between the host and the various molecular virulence attributes of A . pleuropneumoniae. Can J Vet Res, 1990 Apr, 54 Suppl, S28 - 32 Virulence attributes of the liposaccharides of the HAP group organisms; Fenwick BW; The pathogenic potential of purified gram-negative bacterial endotoxin is well documented . However, the role of endotoxin in the disease producing ability of pathogenic organisms, including those of the HAP group, remains in doubt . Endotoxin is not a classic virulence factor, but likely contributes to the pathogenic potential of gram-negative bacteria by activation of host defensive systems which ultimately results in the clinical signs and tissue damage that we recognize as disease . On the other hand, the proinflammatory nature of endotoxin, as well as liposaccharide antigens, play a critical role in the successful elimination of the infecting organism . How the induction of host defensive systems by endotoxin might benefit the infecting organism is difficult to reconcile . In this regard, host response to endotoxin often appears to be in excess of that necessary to clear the infection . Regardless of the role endotoxin might play in the pathogenesis of an infection, liposaccharide antigens are important in the serologically based classification of bacterial strains, are known to contribute to protective immunity, and are useful in the serological diagnosis of infection. J Surg Res, 1990 Apr, 48(4), 349 - 53 Kupffer cell:hepatocyte cocultures release nitric oxide in response to bacterial endotoxin; Billiar TR et al.; Nitric oxide (NO.) is a short-lived intermediate in a biochemical pathway where L-arginine is converted to L-citrulline and nitrite/nitrate (NO2-/NO3-) . This highly reactive molecule is the biologically active component of this inducible pathway in macrophages . Using a rat Kupffer cell:hepatocyte (KC:HC) coculture model, we have previously shown that this combination of cells produces large quantities of both citrulline and NO2-/NO3- if exposed to lipopolysaccharides (LPS) but we did not determine whether nitric oxide was produced or released . We had also shown that this L-arginine metabolism was associated with a profound decrease in total protein synthesis . In these experiments, we show that KC:HC cocultures release nitric oxide into the culture supernatant if exposed to LPS . NO . production by these cells requires L-arginine and is inhibited by NG-mono-methyl-L-arginine . In addition, the time course for NO . release by KC:HC cocultures parallels the previously reported time course for NO2-/NO3- synthesis and the decrease in protein synthesis, supporting the hypothesis that NO . is the reactive nitrogen intermediate of the pathway responsible for this inhibition of protein synthesis . Finally, we show that KC:HC cocultures release more NO . than KC alone in response to LPS, and we propose that the combination of KC and HC acts as a functional unit capable of generating large amounts of NO . from L-arginine in gram-negative sepsis. Am J Physiol, 1990 Apr, 258(4 Pt 1), E678 - 85 Triglyceride kinetics, tissue lipoprotein lipase, and liver lipogenesis in septic rats; Lanza-Jacoby S et al.; The mechanism for the development of hypertriglyceridemia during gram-negative sepsis was studied by examining liver production and clearance of very-low-density lipoprotein (VLDL) triglyceride (TG) . To assess liver output and peripheral clearance the kinetics of VLDL-TG were determined by a constant iv infusion of {2-3H}glycerol-labeled VLDL . Clearance of VLDL-TG was also evaluated by measuring activities of lipoprotein lipase (LPL) in heart, soleus muscle, and adipose tissue from fasted control, fasted E . coli-treated, fed control, and fed E . coli-treated rats . Lewis inbred rats, 275-300 g, were made septic with 8 x 10(7) live E . coli colonies per 100 g body wt . Twenty-four hours after E . coli injection, serum TG, free fatty acids (FFA), and cholesterol of fasted E . coli-treated rats were elevated by 170, 76, and 16%, respectively . The elevation of serum TG may be attributed to the 67% decrease in clearance rate of VLDL-TG in fasted E . coli-treated rats compared with their fasted controls . The suppressed activities of LPL in adipose tissue, skeletal muscle, and heart were consistent with reduced clearance of TG . Secretion of VLDL-TG declined by 31% in livers of fasted E . coli-treated rats, which was accompanied by a twofold increase in the composition of liver TG . Rates of in vivo TG synthesis in livers of the fasted E . coli-treated rats were twofold higher than in those of fasted control rats . Decreased rate of TG appearance along with the increase in liver synthesis of TG contributed to the elevation of liver lipids in the fasted E . coli-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS) Biotechnol Appl Biochem, 1990 Apr, 12(2), 129 - 40 Specific removal of endotoxin from protein solutions by immobilized histidine; Matsumae H et al.; A method for reducing endotoxin contamination in various solutions by immobilized histidine is described . Immobilized histidine is a porous adsorbent suitable for the adsorption of endotoxin with a high affinity over a wide range of pH and temperature and at low ionic strength (gamma/2 less than or equal to 0.1) . When a purified endotoxin originating from Escherichia coli UKT-B was studied, the apparent dissociation constant between endotoxin and the adsorbent was 7.3 X 10(-13) M . The adsorbent was able to remove various kinds of endotoxin originating from gram-negative bacteria; the concentration of endotoxin was reduced from 1000 to less than 0.01 ng/ml in water . It is shown that the adsorbent specifically adsorbs endotoxin provided that the adsorption conditions are properly selected . Some examples of the specific removal of endotoxin from high-molecular-weight physiologically active substances such as tumor necrosis factor and lysozyme are shown. Am J Pathol, 1990 Apr, 136(4), 831 - 41 Mechanisms of tumor necrosis factor-alpha alteration of PMN adhesion and migration; Salyer JL et al.; We have investigated the effects of recombinant human tumor necrosis factor-alpha (rhTNF alpha) on polymorphonuclear leukocytes (PMNs), concentrating on the mechanisms involved in the alterations of PMN-directed migration and adherence by this cytokine . RhTNF alpha profoundly suppressed PMN chemotaxis toward FMLP by 80% . At similar concentrations, it enhanced adhesion to gelatin-coated plastic dishes by more than tenfold and increased the expression of the CD11b antigen to 182% of the control . The monoclonal antibody 60.1, which is directed against the alpha chain of the CD11b/CD18 complex, completely blocked rhTNF alpha, induced inhibition of the chemotactic response to FMLP, and rhTNF alpha induced hyperadherence, suggesting that these effects were related to rhTNF alpha's effects on CD11b antigen expression . The fluid state of the PMN membrane was also decreased by rhTNF alpha . N-butanol, a known membrane fluidizer, partially inhibited the effect of rhTNF alpha on membrane fluidity and chemotaxis and completely reversed its effects on adherence and the expression of the CD11b antigen . Pentoxifylline, an agent that has previously been studied for its ability to prevent some effects of rhTNF alpha on PMNs, completely prevented the effect of rhTNF alpha on chemotaxis, the expression of the CD11b antigen, and membrane fluidity . Pentoxifylline partially prevented changes in adherence caused by this cytokine . Increased CD11b antigen expression caused by rhTNF alpha may result in enhanced PMN adhesion and suppression of migration . These events may, in turn, lead to the accumulation of PMNs on the vascular endothelium, resulting in the extensive vascular and tissue damage that is seen in gram-negative sepsis. J Bacteriol, 1990 Apr, 172(4), 1681 - 7 Mutagenesis of Bordetella pertussis with transposon Tn5tac1: conditional expression of virulence-associated genes; Cookson BT et al.; The Tn5tac1 transposon contains a strong outward-facing promoter, Ptac, a lacI repressor gene, and a selectable Kanr gene . Transcription from Ptac is repressed by the lacI protein unless an inducer (isopropyl-beta-D-thiogalactopyranoside {IPTG}) is present . Thus, Tn5tac1 generates insertion mutations in Escherichia coli with conditional phenotypes because it is polar on distal gene expression when IPTG is absent and directs transcription of these genes when the inducer is present . To test the usefulness of Tn5tac1 in Bordetella pertussis, a nonenteric gram-negative bacterial pathogen, we chose the bifunctional adenylate cyclase-hemolysin determinant as an easily scored marker to monitor insertional mutagenesis . Tn5tac1 delivered to B . pertussis on conjugal suicide plasmids resulted in Kanr exconjugants at a frequency of 10(-3) per donor cell, and nonhemolytic (Hly-) mutants were found among the Kanr colonies at a frequency of about 1% . Of eight independent Kanr Hly- mutants, two were conditional and exhibited an Hly+ phenotype only in the presence of IPTG . Using a new quantitative assay for adenylate cyclase based on high-pressure liquid chromatography, we found that enzymatic activity in these two strains was specifically induced at least 500-fold in a dose-dependent fashion over the range of 0 to 125 microM IPTG . These data show that Ptac serves as a promoter, lacI is expressed and is functional, and IPTG can induce Ptac transcription in B . pertussis . Adenylate cyclase expression in whole cells, culture supernatants, and cell extracts from these strains depended upon IPTG, suggesting that the insertions do not merely alter secretion of adenylate cyclase-hemolysin . Other virulence determinants under control of the vir locus are expressed normally, implying that these Tn5tac1 insertions specifically regulate adenylate cyclase-hemolysin expression . We conclude that Tn5tac1 insertion mutations permit sensitive, exogenous control over the expression of genes of interest, providing a useful tool for studying virulence and other important traits of diverse bacterial species. Z Gastroenterol, 1990 Apr, 28(4), 192 - 7 {The protease inhibitor gabexate mesilate in experimentally-induced acute pancreatitis with early gram-negative infection in the Göttingen minipig: therapeutic efficacy and effects on blood coagulation and fibrinolysis}; Heitz J et al.; Due to its biochemical properties, the newly developed low-molecular protease inhibitor gabexate mesilate is assumed to be efficient in the treatment of complicated acute pancreatitis . This thypothesis was tested using the model of the experimental taurocholate pancreatitis with early artificial E . coli infection in Gottingen mini pigs . Either gabexate mesilate or a placebo was given intravenously 150 min after induction of pancreatitis in a dosage of 2 mg/kg b.w . and h . Median survival time was 15 h in the gabexate mesilate treated animals (n = 7) as compared to 34 h in the placebo group (n = 7); this difference was not significant . The application of gabexate mesilate had no influence on the increased coagulation activity (decrease of prothrombin time, antithrombin III and platelet count) nor on the additional hyperfibrinolysis with concomitant decrease of plasminogen and antiplasmin . Prothrombin time and antithrombin III were less distinctly decreased in the placebo group; decrease of platelet count was more pronounced . On the basis of this study the hypothesis is not confirmed that treatment with gabexate mesilate for necrotizing experimental pancreatitis with additional gram-negative infection has a therapeutic efficiency. J Immunol, 1990 Apr 1, 144(7), 2566 - 71 CD18-deficient cells respond to lipopolysaccharide in vitro; Wright SD et al.; The CD11/CD18 complex of leukocyte adhesion molecules has been shown to bind LPS on the surface of gram negative bacteria and LPS-coated erythrocytes (J . Exp . Med . 164:1876, 1986) . LPS elicits several responses in leukocytes including secretion of TNF-alpha and IL-1 beta, and priming for enhanced release of oxygen radicals such as superoxide anion . To determine if expression of CD18 molecules is necessary for these effects of LPS, we have examined the responses of leukocytes from CD18-deficient patients . Three of the patients in this study are characterized for the first time here, and three were described elsewhere . Monocytes and macrophages from CD18-deficient patients synthesized normal amounts of IL-1 beta and TNF-alpha in response to LPS . Further, PMN and monocytes from CD18-deficient patients showed normal priming for enhanced release of superoxide anion in response to LPS . Although a small contribution of CD18 molecules to some responses cannot be ruled out by our data, we may conclude that CD18 molecules are not essential for cellular responses to LPS. Obstet Gynecol, 1990 Mar, 75(3 Pt 1), 402 - 6 A clinical and microbiologic analysis of risk factors for puerperal endometritis; Newton ER et al.; Predictors of postpartum endometritis were identified in 607 asymptomatic, laboring women . One hundred (16.5%) developed postpartum endometritis . Multivariate analysis using stepwise logistic regression identified cesarean delivery (relative risk 12.8; P less than .0001) as the dominant overall predictor . In patients with cesarean delivery (N = 124), prophylactic antibiotics (relative risk 0.54; P less than .0002) and high-virulence bacteria or Mycoplasma hominis (relative risk 1.4; P less than .01) predicted the incidence of endometritis, and in patients with vaginal delivery (N = 483), "bacterial vaginosis organisms" (relative risk 14.2; P less than .001) and aerobic gram-negative rods (relative risk 4.2; P less than .01) predicted endometritis . Despite significant associations found on univariate analysis, clinical variables such as duration of labor, rupture of membranes, and internal monitoring were not predictive of endometritis in the multivariate analysis . Our findings show that cesarean delivery and certain organisms, such as bacterial vaginosis or high-virulence organisms, predict endometritis, and that clinical variables may be facilitators rather than predictors of endometritis. Am J Emerg Med, 1990 Mar, 8(2), 152 - 61 Modern approaches to the therapy of septic shock; Putterman C; Bacteremia from gram-negative rods is a great cause of concern for hospital physicians today . Shock-complicating gram-negative sepsis has a mortality rate of 60% and above, despite early diagnosis and treatment . Intensive research efforts have shown new pathophysiological mechanisms and mediators involved in septic shock, with changes in recommended treatment protocols . In this report, the authors review the use of corticosteroids, fibronectin, naloxone hydrochloride, and immunotherapy, with emphasis on theoretical considerations and relevant clinical experience . Although these treatment methods may have been promising initially, data from large double-blind human trials are either lacking or unencouraging . While continued research and modern therapeutic approaches should improve future survival rates from septic shock, use of the therapies reviewed should be considered experimental at this time. Int J Food Microbiol, 1990 Mar, 10(2), 157 - 76 A simple nutrient-tolerance (NT) test for the characterization of the different types of oligocarbotolerant and oligocarbophile water bacteria from non-carbonated mineral water; Schmidt-Lorenz W et al.; Comparative determination of the specific growth kinetics in mineral water and low and higher concentrated broths at 20 degrees C of 25 selected Gram-negative bacteria isolated from natural non-carbonated mineral water yielded three groups: (1) facultative oligocarbotolerants--with faster growth in normal broth (In g l-1: yeast extract 2.5; casein peptone 5.0; glucose 1.0); (2) obligate oligocarbotolerants--with equal rates of growth in normal and 1:10 diluted broth; and (3) oligocarbophiles--with faster growth in 1:10 diluted broth and in mineral water . In addition, three nutrient types, 'eu-, meso- and oligotrophic' could be distinguished on the basis of full, weak and no growth in brain-heart infusion broth . Further characterization was made between slow and very slow growth types in 1:10 diluted broth . All 25 isolates were psychrotrophic with a minimum growth temperature below 0 degree C . The optimum and maximum temperatures of growth in 1:10 diluted broth, as determined in a temperature gradient incubator were between 20 and 32, and between 29 and 34 degrees C with an average of 26 and 31 degrees C, respectively . Based on these results a very simple nutrient-tolerance test was proposed . After inoculation of the three media, 1:10 diluted broth, normal broth and brain-heart infusion, it is only necessary to check whether or when visible turbidity occurs during 2 weeks incubation at 20 degrees C . This allows additional characterization of bacteria from natural mineral water, which are often difficult to identify, on the basis of growth characteristics in various types of nutrient media. J Gen Microbiol, 1990 Mar, 136 ( Pt 3), 589 - 98 Molecular analysis of beta-lactamases from four species of Streptomyces: comparison of amino acid sequences with those of other beta-lactamases; Forsman M et al.; Genes encoding extracellular beta-lactamases of Streptomyces badius, Streptomyces cacaoi, Streptomyces fradiae and Streptomyces lavendulae were cloned and mapped in Streptomyces lividans . DNA sequence analysis of the beta-lactamase genes revealed a high overall G + C content, ranging from 71 to 75 mol%, with a G + C content of 95 mol% at the third position of the codons for all four genes . The primary structure of the beta-lactamases including their signal peptides was deduced . The four beta-lactamases exhibited homology to each other and to class A beta-lactamases from other bacterial genera . We suggest that Streptomyces beta-lactamases are representatives of a superfamily of genes, from which class A beta-lactamases of Gram-negative bacteria may have evolved. J Gen Microbiol, 1990 Mar, 136 ( Pt 3), 441 - 6 Taxonomic studies on methylotrophic bacteria by 5S ribosomal RNA sequencing; Bulygina ES et al.; Nucleotide sequences of 5S ribosomal RNA (rRNA) isolated from 19 strains of Gram-negative methylotrophic bacteria were determined . Comparison of these sequences allowed construction of a tentative phylogenetic tree and showed that the bacteria analysed belong to the Proteobacteria and fell into several clusters, including obligate methanotrophs, obligate methylotrophs and several groups of facultative methylotrophs . Taxonomic relations between methylotrophic and non-methylotrophic bacteria are discussed, and the polyphyletic nature of methylotrophy as a taxonomic feature is highlighted. J Pharm Sci, 1990 Mar, 79(3), 261 - 5 Simultaneous derivative spectrophotometric determination of aztreonam and L-arginine in injections; Morelli B; Mixtures of aztreonam, 2S-{2 alpha,3 beta(Z)}-2-{{{1-(2-amino-4-thiazolyl-2- {(2-methyl-4-oxo-1-sulfo-3-azetidinyl) amino}-2-oxo- ethylidine}amino}oxy}-2-methyl-propanoic acid, and L-arginine are assayed by peak-to-baseline, peak-to-peak, and zero-crossing second-derivative spectrophotometry . Aztreonam is the first of a new class of beta-lactam antibiotics (i.e., the mono-bactam antibiotics, highly effective against aerobic gram-negative bacteria) . Beer's law is followed for up to 31 micrograms/mL of aztreonam and L-arginine in the presence of one another . Detection limits at the p = 0.05 level of significance range from 0.11 to 0.37 micrograms/mL . The method was successfully applied to laboratory mixtures and commercial injections containing these substances. Zhonghua Min Guo Xiao Er Ke Yi Xue Hui Za Zhi, 1990 Mar-Apr, 31(2), 110 - 5 {Bacterial meningitis in neonates}; Wu SF et al.; A retrospective study of the clinical features and management of 27 newborns with meningitis is presented . All were treated in the China Medical College Hospital between July 1981 and July 1988 . Seven were premature and twenty were full term . There were 11 males and 16 females . Five infants had low birth weight (less than 2500 g) and 12 cases were diagnosed in the first week of life (early onset) . E.coli was the predominant pathogen . The most common presenting manifestations were nonspecific in nature-fever, poor feeding, respiratory distress and poor activity, etc.. . Six patients died during the period of acute illness and the case fatality rate was 22.2% . Of the 21 patients who survived, six had sequelae (four hydrocephalus, one subdural effusion, one microcephaly), giving an over-all rate of 19 percent in the survivors . The presence of gram-negative bacterial infection and high protein content in CSF correlated significantly with the higher mortality rate. Pediatr Dermatol, 1990 Mar, 7(1), 11 - 8 Cat-scratch disease: a review of the literature; Shinall EA; Cat-scratch disease (CSD) is a common cause of chronic lymphadenopathy (especially regional) that primarily affects children and adolescents . The clinical diagnosis of CSD is based on the presence of three of four criteria, which may include a positive CSD skin test . Usually a benign, self-limiting disease, CSD may sometimes have atypical manifestations and serious complications, particularly in immunocompromised hosts . Cat-scratch disease is now known to be caused by a small, gram-negative, pleomorphic bacterium . Antibiotics are dramatically effective against CSD in immunocompromised patients, but are not {corrected} of proven benefit in typical cases . Most patients recover with only symptomatic treatment . This article reviews the history of CSD research, clinical features of typical and atypical CSD, and current topics of interest in CSD research, especially in the areas of diagnosis and treatment. World J Surg, 1990 Mar-Apr, 14(2), 196 - 203 Intraabdominal infection: pulmonary failure; Runcie C et al.; Since its first description in 1967, the mortality of the adult respiratory distress syndrome (ARDS) has remained unchanged despite the increasing sophistication of supportive techniques . Few patients now die of refractory hypoxemia, the majority succumbing to the multiple systems organ failure syndrome, commonly due to sepsis . Sepsis is both the most common cause of ARDS, usually involving the abdomen, and the most frequent complication, usually affecting the lungs . ARDS is, thus, increasingly seen as the pulmonary component of multiple systems organ failure, triggered by the systemic response to sepsis . In critically ill patients, impairment of hepatic function and of the barrier function of the gut mucosa allows translocation of endotoxin derived from the aerobic Gram-negative bacteria within the gut . This releases mediators which are responsible for the activation of cellular and humoral cascades, resulting in the pathological changes seen in ARDS . This sequence of events underlines the importance of therapies directed at abnormal colonization of the gastrointestinal tract and elimination of the gut endotoxin pool . Selective decontamination of the digestive tract is attractive in that it attacks the problem from 2 sides: first, by eliminating colonization, it appears effective in preventing secondary infection and, second, it may also play a role in reducing the enteric endotoxin pool . Recent descriptions of pathological oxygen supply dependency in both ARDS and septic patients emphasize the similarity of pathophysiological abnormalities in the 2 conditions . Intensive supportive therapy to achieve adequate oxygen transport and aggressive investigation and surgical management of septic foci are the cornerstones of management of the established syndrome. J Med Microbiol, 1990 Mar, 31(3), 213 - 8 Identification and cloning of the type IIIa plasmid-encoded dihydrofolate reductase gene from trimethoprim-resistant gram-negative bacteria isolated in Britain; Thomson CJ et al.; A clinical strain of Escherichia coli isolated in Nottinghamshire in 1980 was shown to harbour the type IIIa trimethoprim-resistant dihydrofolate reductase gene, previously identified on only one occasion, in New Zealand in 1979 . The gene was identified by hybridisation with an 855-bp type III gene probe and its classification as a type IIIa dihydrofolate reductase was confirmed by detailed biochemical analysis of the enzyme product . The dihydrofolate reductase was identical in size and isoelectric point with the original type IIIa enzyme and shared similar inhibitory and kinetic profiles . The trimethoprim resistance gene was subsequently cloned and the type IIIa dihydrofolate reductase gene was localised to a 700-bp EcoRI-PstI fragment . This smaller fragment may prove to be a more specific DNA probe for the future identification of type IIIa dihydrofolate reductase genes. Circ Shock, 1990 Mar, 30(3), 265 - 78 Effect of gram-negative endotoxin on levels of serum corticosterone, TNF alpha, circulating blood cells, and the survival of rats; Feuerstein G et al.; The relationship between serum tumor necrosis factor (TNF alpha), circulating blood cells, plasma corticosterone (CS), and survival was studied in conscious rats injected intravenously with E . coli lipopolysaccharide (LPS, 0.0001-28.8 mg/kg) . The TNF alpha response was dose-related, peaked at 90-120 min after LPS injection, and subsided 6 hr later . The CS response showed an earlier onset, prolonged (greater than 12 hr) duration, and a broader dose-response pattern . White blood cells (WBC) and platelet depletion peaked at 2-4 and 24-48 hr post-LPS, respectively; however, no changes in the cell count were observed at LPS doses which produced greater than 50% maximal increase in TNF alpha levels . Survival curves for each of the LPS doses were significantly different from the doses of LPS which elicited TNF alpha release . Systemic injection of human recombinant TNF alpha (h-rTNF alpha), which produced plasma levels of TNF alpha 10 x 10(7) U/ml, much higher than the maximal TNF alpha levels produced by LPS, 1-2 x 10(5) U/ml, failed to affect survival, plasma WBC, or hematocrit . Our data suggest that factors other than TNF alpha produced by high doses of LPS are essential in eliciting LPS-induced death. Am Rev Respir Dis, 1990 Mar, 141(3 Pt 2), S145 - 50 Receptors in airway disease . Beta-adrenoceptors in lung inflammation; Nijkamp FP et al.; Beta-adrenoceptor dysfunction and increase in airway reactivity can be induced by administration of gram-negative bacteria, endotoxin, viruses, and allergens in laboratory animals . However, the deterioration of lung beta-adrenoceptor function is not invariably associated with lung inflammation . Severe asthmatics, but not asthmatics per se, show a diminished beta-adrenoceptor function of airway smooth muscle . These changes are probably a consequence of the active disease state rather than an intrinsic component of asthma . Mediators released from inflammatory cells such as reactive oxygen species and fatty acid metabolites may directly or indirectly induce beta-adrenoceptor dysfunction . Beta-adrenoceptor function of leukocytes from asthmatic patients can be decreased as well and it is suggested that lymphokines like interleukin-2 and interferon-gamma may affect beta-adrenoceptor function . A disturbed beta-adrenoceptor function on inflammatory cells themselves may have consequences for their immune function, mediator release, and effect on surrounding tissues. Ugeskr Laeger, 1990 Feb 5, 152(6), 379 - 81 {Sinusitis caused by nasotracheal intubation}; Pedersen J et al.; Sinusitis following nasotracheal intubation (NTI) is an almost overlooked complication . The pathogenesis is believed to be development of oedema of the nasal mucosa due to irritation by the tube in the nasal cavity . The size of the tube is probably not important, whereas the duration of intubation is of major importance . The infection is often caused by Gram-negative microorganisms . The frequency is found to be 0.3% in short term intubation (less than 5 days) and 40.4% in long term intubation (greater than 5 days) . In neurosurgical patients treated with NTI sinusitis is found in 52% and 100% of the patients . In adults the indications for prolonged NTI instead of orotracheal intubation and early tracheostomy should be considered carefully . If fever or sepsis develops in patients treated with NTI, investigations for sinusitis should be included in the search for focus . When sinusitis has developed, the tube should be removed, and the patient tracheostomised . If this treatment fails, surgical drainage should be performed. Epidemiol Infect, 1990 Feb, 104(1), 39 - 45 Prevalance of antibodies to 15 antigens of Legionellaceae in patients with community-acquired pneumonia; McIntyre M et al.; Sera from 252 patients with community-acquired pneumonia were examined for the presence of antibodies to 15 antigens of 7 Legionella spp . by indirect immunofluorescent antibody testing . The sera had been collected as part of the British Thoracic Society/Public Health Laboratory Service study of community-acquired pneumonia in adults . We also examined sera from 20 patients with gram-negative sepsis . Using a limited range of antigens of L . pneumophila, nine cases of legionellosis were diagnosed in the original study . However, using antigens to other Legionella spp., we identified two further cases, caused by L . micdadei and L . gormanii respectively . Twenty-six other patients had titres of 16 or 32 to one or more antigens, most commonly L . bozemanii serogroup 1, L . micdadei and L . dumoffi . None of the patients with non-legionella pneumonia, however, had significant changes in legionella antibody titres . All of the patients with Gram-negative sepsis had titres of less than 16. J Dairy Res, 1990 Feb, 57(1), 33 - 9 Effect of cleaning-in-place disinfectants on wild bacterial strains isolated from a milking line; Mattila T et al.; Two cleaning-in-place disinfectant systems (alkaline-acid, alkaline-Cl2) were tested on wild bacteria isolated from a milking line . Most of the isolated strains were Gram-negative and capsule-positive . The effect of milk and cream on the resistance of the wild strains to the cleaning systems was tested . Suspension tests did not show the resistance of bacteria whereas the surface test showed high resistance towards all cleaning-agents . Milk had a protective effect only on the surfaces whereas cream showed mild protection in suspension and complete protection on the surfaces . It was concluded that the growth pattern of contaminants on surfaces needs further attention and the laboratory suspension tests do not correlate with the practical situation . Disinfectant tests based on bacterial growth on the surface are of importance. J Clin Microbiol, 1990 Feb, 28(2), 400 - 1 Ocular infection caused by Psychrobacter immobilis acquired in the hospital; Gini GA; The name Psychrobacter immobilis recently has been proposed for a group of chiefly psychrotrophic, aerobic, gram-negative, nonmotile, oxidase-positive coccobacilli commonly found associated with fish, processed meat, and poultry products . This article reports an ocular infection in a 12-day-old newborn who acquired the infection in the hospital, probably because of frequent manipulations in a closed aerated incubator . Also, this report intends to alert microbiologists to opportunistic infections that might be confused with infections caused by unusual oxidase-positive, gram-negative diplococci and to the isolation of P . immobilis from a human infection. Gut, 1990 Feb, 31(2), 139 - 43 Tightly spiral shaped bacteria in the human stomach: another cause of active chronic gastritis? Morris A, Ali MR, Thomsen L, Hollis B. Tightly spiral shaped Gram negative bacteria were seen in the gastric biopsies obtained from two patients undergoing gastroscopy . Active chronic gastritis was present in both patients and one patient also had gastric ulceration . Attempts to culture the organism by a number of methods were unsuccessful but positive urease results were obtained in both patients . Both patients were treated with colloidal bismuth subcitrate . Biopsies taken after treatment showed resolution of infection and histological gastritis . These results suggest that Helicobacter pylori is not the only organism associated with chronic active gastritis in man. J Bone Joint Surg Am, 1990 Feb, 72(2), 272 - 8 Two-stage reimplantation for the salvage of total knee arthroplasty complicated by infection . Further follow-up and refinement of indications; Windsor RE et al.; Thirty-eight total knee replacements (in thirty-five patients) that were complicated by infection were treated with a two-stage protocol for reimplantation . The clinical results in these knees (nine of which have been previously reported on) were evaluated at an average follow-up of four years (range, 2.5 to ten years) . There was only one documented recurrence of infection with the original organism . Three patients in whom the immunological system was suppressed had a subsequent hematogenous infection with a different organism . According to the knee-rating system of The Hospital for Special Surgery, there were eleven excellent, thirteen good, six fair, and seven poor results . For one patient who had severe polyarticular rheumatoid arthritis, the result could not be rated . The results of this study suggested that the two-stage protocol for reimplantation, with a six-week interval of intravenous antibiotic therapy, is the procedure of choice for the treatment of an infection around a total knee arthroplasty . A patient who has polyarticular rheumatoid arthritis and in whom the immunological system is suppressed may not be an ideal candidate for the protocol . Gram-negative bacterial infection may be treated with this protocol, provided the organism is sensitive to relatively non-toxic antibiotic medication. J Biochem (Tokyo), 1990 Feb, 107(2), 190 - 6 Primary structure of a fucose-specific lectin obtained from a mushroom, Aleuria aurantia; Fukumori F et al.; Aleuria aurantia lectin (AAL) is a protein composed of two identical subunits having no carbohydrate chain and shows sugar-binding specificity for L-fucose . Full-length cDNA encoding for the lectin has been isolated from a lambda gt11 library, screened with an antiserum directed against AAL . The cDNA clone contained 1,370 nucleotides and an open reading frame of 939 nucleotides encoding 313 amino acids . The amino-terminal sequence (residues 1-30) of the lectin isolated from the mushroom coincided with the deduced amino acid sequence starting from proline at the 2nd residue, indicating that the mature AAL consists of 312 amino acids . Its molecular weight is calculated to be 33,398 . The deduced amino acid sequence shows that AAL includes six internal homologous regions, and has considerable homology with a hemagglutinin from a Gram-negative bacterium, Myxococcus xanthus, which forms a fruiting body . No significant homology was observed with higher plant or animal lectins . The recombinant AAL produced by Escherichia coli JM109 carrying the AAL expression plasmid pKA-1 {Fukumori, F . et al . (1989) FEBS Lett . 250, 153-156} was purified from the cell lysate by affinity chromatography using a fucose-starch column, and hundreds of milligrams of the lectin was obtained . The recombinant lectin showed the same biochemical characteristics and sugar binding specificity as did the natural AAL. Proc Natl Acad Sci U S A, 1990 Feb, 87(3), 942 - 5 Two independent retrons with highly diverse reverse transcriptases in Myxococcus xanthus; Inouye S et al.; A reverse transcriptase (RT) was recently found in Myxococcus xanthus, a Gram-negative soil bacterium . This RT has been shown to be associated with a chromosomal region designated a retron responsible for the synthesis of a peculiar extrachromosomal DNA called msDNA (multicopy single-stranded DNA) . We demonstrate that M . xanthus contains two independent, unlinked retrons, one for the synthesis of msDNA-Mx162 and the other for msDNA-Mx65 . The structural analysis of the retron for msDNA-Mx65 revealed that the coding regions for msdRNA (msr) and msDNA (msd), and an open reading frame (ORF) downstream of msr are arranged in the same manner as found for the Mx162 retron . The ORF encodes a polypeptide of 427 amino acid residues . The amino-terminal domain (residues 1-138) shows no striking similarity to these proteins presently available in the data bases including the msDNA-Mx162 ORF, while the sequence from residues 139-394 can be aligned with various known RT sequences and has 47% identity with the RT domain of the msDNA-Mx162 ORF . On the basis of these findings, possible origins of two highly diverse retrons on the M . xanthus chromosome are discussed. J Immunol Methods, 1990 Jan 24, 126(1), 109 - 17 Increased resolution of lipopolysaccharides and lipooligosaccharides utilizing tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis; Lesse AJ et al.; We utilized the recently described tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (TSDS-PAGE) system to study the lipooligosaccharides (LOS) and lipopolysaccharides (LPS) of gram negative bacteria . TSDS-PAGE resulted in a high degree of resolution of LOS and LPS in the 'mini-gel' format . TSDS-PAGE resulted in the LOS and LPS migrating as a function of their Mr during electrophoresis and allowed estimation of Mr from a protein standard . Several species of LOS were analyzed . The newly described procedure allowed a more rapid and accurate analysis of LOS and the core region of LPS. J Biol Chem, 1990 Jan 5, 265(1), 190 - 4 Molecular characterization of an anion pump . The ArsB protein is the membrane anchor for the ArsA protein; Tisa LS et al.; R-factor mediated bacterial resistance to arsenical salts occurs by active extrusion of the toxic oxyanions from cells of gram negative bacteria . The ars operon of the conjugative plasmid R773 encodes an anion pump . The pump has two polypeptide components . The catalytic subunit, the ArsA protein, is an oxyanion-stimulated ATPase . The membrane component, the ArsB protein, has been localized in the inner membrane of Escherichia coli . The ArsA and ArsB proteins have been postulated to form a membrane complex which functions as an anion-translocating ATPase . In this study evidence is presented showing that expression of the arsB gene is required to anchor the ArsA protein to the inner membrane . Binding studies with purified ArsA to membranes with and without the arsB gene product confirm this requirement . Membranes of uncA mutants containing both the ArsA and ArsB proteins exhibit arsenite(antimonite)-stimulated ATPase activity . These results support the model in which the ArsA protein is the catalytic energy transducing component of the anion pump, whereas the integral membrane ArsB protein serves as both the anion channel and membrane binding site for the ArsA protein. J Vet Intern Med, 1990 Jan-Feb, 4(1), 17 - 25 Immunity targeting common core antigens of gram-negative bacteria; Tyler JW et al.; Antibodies against common gram-negative core antigens can provide protection against environmental, commensual, and contagious bacteria that cause life-threatening gram-negative sepsis/endotoxemia . Cross-protective immunity may be effective against many common livestock diseases, including neonatal coliform septicemia, coliform mastitis, septic metritis, and severe enteritis . The greatest potential benefit of cross-protective immunity may be realized in the reduction of ill-thrift and mortality of neonates . The use of active and passive immunization against core antigens common to gram-negative bacteria has been successful in human beings, laboratory animals, and livestock . Protection has been observed following challenge with either heterologous endotoxin or unrelated, live, virulent gram-negative bacteria. Circ Shock, 1990 Jan, 30(1), 5 - 13 Does the heart fail in endotoxin shock? Abel FL. Evidence for a major participatory role of the myocardium in the hemodynamic response to endotoxemia and sepsis has been controversial . Early interpretations about in situ changes in cardiac performance during shock were confounded by the concomitant influence of uncontrolled pathophysiologic adjustments in cardioactive variables such as preload, afterload, sympathoadrenal discharge, left ventricular diastolic compliance (stiffness), and in some studies myocardial hypoperfusion/ischemia . Despite such complexities, many recent studies with intact subjects and isolated preparations consistently point toward a relatively early involvement of myocardial dysfunction in circulatory shock syndromes associated with gram-negative bacteria and sepsis . The cardiac dysfunction cannot be accounted for by a direct toxic effect of the endotoxin molecule itself on cardiac myocytes . Rather, evidence is compelling that endotoxin interacts first with other types of cellular or tissue elements . The latter evoke biologic reactions through some unknown pathway culminating in deleterious effects in the heart . Discovery of the casual mechanism(s) responsible for myocardial sequelae of endotoxemia and sepsis remains a key objective in experimental and clinical research in circulatory shock pathogenesis. Am J Kidney Dis, 1990 Jan, 15(1), 1 - 7 Abdominal catastrophes and other unusual events in continuous ambulatory peritoneal dialysis patients; Steiner RW et al.; Primary bacterial peritonitis and catheter-associated infections compose the large majority of abdominal events in continuous ambulatory peritoneal dialysis (CAPD) patients . Yet occasionally primary pathology involving the abdominal viscera develops, and surgery is frequently considered . The early manifestations of intraabdominal inflammation or bleeding in patients undergoing CAPD depend on the pathological process, its access to the peritoneal cavity, and whether generalized bacterial peritonitis supervenes to obscure helpful physical findings . Clear dialysate is not a reliable sign that major pathology is absent, nor does initial stabilization of the clinical course with antibiotic therapy uniformly indicate that surgery will not be necessary . Polymicrobial peritonitis may develop in cholecystitis, pancreatitis, or from a colonic source, the latter featuring more bacterial species and more gram-negative and anaerobic organisms . A history directed at progression of symptoms and sites of abdominal discomfort and an examination for deep local tenderness and bowel incarcerated in an abdominal wall hernia are essential . Measurement of dialysate amylase and Gram stain of dialysate for food fibers may be helpful . Imaging techniques such as abdominal radiographs for dilated bowel or free subdiaphragmatic air, ultrasonography of the gallbladder or pancreas, computed tomographic (CT) scanning of the lower abdomen, and water-soluble contrast colonic studies may help identify the pathologic process . Special studies such as these should be considered early in the course of suspected unusual abdominal events in patients on CAPD. J Leukoc Biol, 1990 Jan, 47(1), 70 - 8 An endotoxin-induced factor distinct from interleukin-1 and tumour necrosis factor alpha produced by the THP-1 human macrophage line stimulates polymorphonuclear leukocyte infiltration in vivo; Megyeri P et al.; Endotoxin and gram-negative bacteria induce vigorous inflammatory reactions . Our previous work showed that rabbit macrophages (M phi) incubated with endotoxin produce a 45,000 dalton protein that recruited polymorphonuclear leukocytes (PMNL) into the skin of rabbits . This factor was separated from interleukin-1 (IL-1) but could not be unequivocally distinguished from rabbit tumour necrosis factor (TNF alpha) . Here we have examined the human M phi cell line, THP-1, for the production of an analogous protein . After exposure to phorbol diester the THP-1 cells assumed the characteristic M phi phenotype and function . During 6 hours of culture with LPS these M phi released a factor(s) that caused PMNL recruitment into the skin of rabbits when injected intradermally, measured using 51Cr-labelled blood leukocytes . This activity, referred to as PMNL recruiting activity (PRA), was heat labile, and its production was blocked by cycloheximide, suggesting that this is most likely a de novo synthesized protein . Sephadex-G 100 and Superose-12 FPLC chromatography indicated a molecular weight in the 45,000-65,000 dalton range . The active fractions were free of IL-1 activity (less than 0.2 U/ml), and Superose-12 chromatography separated the peak of PRA, which eluted around 45,000 daltons, from TNF alpha eluting at 20,000 daltons . The peak PRA was not neutralized by antiserum to IL-1 alpha, IL-1 beta TNF alpha, IL-6, and granulocyte-macrophage colony-stimulating factor (GMCSF), indicating that it was distinct immunologically from these cytokines . The major PRA did not induce migration of rabbit or human PMNLs in vitro in a Boyden chamber chemotaxis assay, although peaks of chemotactic activity and weak PMNL recruitment in vivo were detected in fractions eluting around 15,000 daltons and 800 daltons . The generation of PRA by a human M phi cell line is analogous to that reported previously with rabbit M phi . Here we extend these observations to a human M phi system and confirm that this molecule is distinct from several other M phi cytokines and M phi chemotactic factors with inflammatory properties. Infect Immun, 1990 Jan, 58(1), 222 - 7 The capsular polysaccharide is a major determinant of serum resistance in K-1-positive blood culture isolates of Escherichia coli; Leying H et al.; Serum resistance is a major virulence factor of gram-negative bacteria, and K-1 polysaccharide has been shown to contribute to serum resistance in selected strains . To obtain further information about the role of K-1 in serum resistance and to find out whether loss of the ability to produce K-1 can induce loss of serum resistance, we studied the serum resistance of mutants derived from completely serum-resistant, K-1-positive blood culture isolates of Escherichia coli by selection for resistance to infection with K-1 specific bacteriophages . The amounts of K-1 polysaccharide produced by wild-type strains and mutants were measured, and outer membrane protein and lipopolysaccharide (LPS) patterns were analyzed . In each group of mutants, several highly serum-sensitive strains were found . All mutant strains expressed less K-1 than did the corresponding wild-type strains . Mutants that became highly serum sensitive always had less K-1 than did mutants with less-pronounced changes of serum resistance . A few mutants derived from different wild-type strains showed increased expression of outer membrane proteins with molecular weights of about 46,000 and 67,000 . All of the wild-type strains examined had smooth-type LPS, and only two mutants had altered LPS structures; alterations of mutants in outer membrane proteins and LPS could not be correlated with alterations of serum resistance . The results indicate that for K-1-positive blood culture strains of E . coli, K-1 expression is a prerequisite for serum resistance, and loss of ability to synthesize K-1 leads to loss of serum resistance. Cancer, 1990 Jan 1, 65(1), 9 - 16 Beta-lactam regimens for the febrile neutropenic patient; Bodey GP et al.; A total of 535 evaluable febrile episodes in neutropenic patients were randomly assigned to treatment with ticarcillin-clavulanate plus vancomycin (TV), ceftazidime plus vancomycin (CV), or all three antibiotics (TCV) . The TCV regimen was significantly more effective than TV, considering all evaluable episodes, documented infections, gram-negative infections, and infections in patients with persistent severe neutropenia (less than 100 neutrophils/mm3) . The results with CV were intermediate between TV and TCV . The toxicities were similar with all three regimens and consisted primarily of skin rashes . The TCV regimen is effective for empiric therapy of fever in neutropenic patients and probably should be utilized in preference to CV or TV, although its superiority over CV in this study was inconclusive. Arch Microbiol, 1990, 154(2), 192 - 8 Microbial degradation of acrylamide monomer; Shanker R et al.; Acrylamide, a neurotoxic monomer with extensive industrial applications was found to be degraded by the microorganisms present in a tropical garden soil . A bacterium capable of degrading acrylamide was isolated from this soil by enrichment . It was found to be aerobic, gram-negative, motile, short rod and identified as Pseudomonas sp . The bacterium degraded high concentrations of acrylamide (4 g/l) to acrylic acid and ammonia which were utilized as sole carbon and nitrogen source for growth . An amidase was involved in the hydrolysis of acrylamide, which could act on other short chain amides like formamide and acetamide but not on acrylamide analogues: methacrylamide and N,N-methylene bisacrylamide . The enzyme was sensitive to catabolite repression by succinate both in presence as well as absence of nitrogen source. Infection, 1990, 18 Suppl 1, S19 - 21 The incidence of gram-negative bacterial and fungal infections in liver transplant patients treated with selective decontamination; Wiesner RH; We have used selective bowel decontamination in 145 consecutive patients undergoing liver transplantation . In these 145 patients undergoing initial transplantation, we have experienced 42 major bacterial infections in 37 patients . Only five patients have experienced a gram-negative infection and only 2 of 26 documented bacteremias were associated with gram-negative organisms . To date, we have experienced only one systemic Candida infection . These results suggest that selective bowel decontamination may play a major role in reducing the previously reported high incidence of gram-negative and Candida sepsis following liver transplantation. Plasmid, 1990 Jan, 23(1), 80 - 4 Initiation of DNA synthesis in the transfer origin region of RK2 by the plasmid-encoded primase: detection using defective M13 phage; Yakobson E et al.; The broad host range IncP (IncP1) plasmids of gram-negative bacteria encode DNA primases that are involved in conjugal DNA synthesis . The primase of RK2/RP4 is required for efficient DNA transfer to certain gram-negative bacteria, indicating that the enzyme primes complementary strand synthesis in the recipient . In vitro, the primase initiates synthesis of oligoribonucleotides at 3'-dGdT-5' dinucleotides on the template strand . In this report, replication-defective M13 phage are used to assay the ability of the RK2-encoded primase to initiate complementary strand synthesis in vivo on single-strand templates containing the RK2 origin of conjugal transfer (oriT) or the RK2 origin of vegetative replication (oriV) . The results show that sequences from either strand of the oriT region serve as efficient substrates for the RK2 primase and can enhance the growth of the defective M13 vectors delta E101 and delta Elac to levels approaching wild-type . The primise-oriT interaction appeared specific, since neither the oriV sequence nor another RK2 region, trfB, significantly enhanced growth of the defective phage, either in the presence or in the absence of the primase . In contrast to ColEl and F, this study also shows that the oriV region of RK2 lacks sites that are recognized by the host-specified DNA priming systems . The results suggest that the oriT region contains sites on both DNA strands that are efficient substrates for the plasmid-encoded primase, facilitating initiation of complementary strand DNA synthesis in both donor and recipient during conjugation. Arch Microbiol, 1990, 153(4), 360 - 7 Characterization of Xanthobacter strains H4-14 and 25a and enzyme profiles after growth under autotrophic and heterotrophic conditions; Meijer WG et al.; All Xanthobacter strains studied are versatile autotrophic bacteria, able to grow on methanol and other substrates . Strain 25a, a yellow-pigmented, pleomorphic, Gram-negative bacterium, capable of autotrophic growth on methanol, formate, thiosulfate, and molecular hydrogen, was isolated from an enrichment culture inoculated with soil from a subtropical greenhouse . Subsequent studies showed that the organism also grows on a wide range of multicarbon substrates . Ammonia, nitrate and molecular nitrogen were used as nitrogen sources . The taxonomic relationship of strains H4-14 and 25a with previously described Xanthobacter strains was studied by numerical classification . Strain H4-14 was identified as a X . flavus strain, but the precise position of strain 25a remained uncertain . It probably belongs to a new species of the genus Xanthobacter . The levels of various enzymes involved in autotrophic and heterotrophic metabolism were determined following growth of strains H4-14 and 25a in batch and continuous cultures . The mechanisms involved in controlling ribulose-1,5-bisphosphate carboxylase/oxygenase synthesis in Xanthobacter strains appear to be comparable to those observed for other autotrophic bacteria, namely repression by organic compounds and derepression by autotrophic energy sources, such as methanol and hydrogen. Mol Gen Mikrobiol Virusol, 1990 Jan, (1), 22 - 6 {Functional properties of the pOV13 plasmid as a vector for DNA cloning in a broad spectrum of gram negative bacteria}; Zakharenko VI et al.; The birepliconed plasmid pOV13 possesses all the properties of a vector for DNA cloning in a broad host range of bacterial cells . pOV13 is transfered by transformation and stably inherited by Escherichia coli, Brucella, Pseudomonas cells determining the resistance to streptomycin, tetrocycline and kanamycin in these bacteria . The plasmid pOV13 is a multicopy plasmid optimal in replication capacity (23kb) . The plasmid carries single sites for some restriction endonucleases that are used for DNA cloning, including some restriction sites in antibiotic resistance genes . The examples of DNA cloning with the selection of recombinant clones by the insertional inactivation of kanamycin or tetracycline resistance and expression of the cloned DNAs are presented. Clin Pharm, 1990 Jan, 9(1), 40 - 4 Expanded gentamicin volume of distribution in patients with indicators of malnutrition; Zarowitz BJ et al.; Visceral protein, body weight, and body composition were assessed in patients receiving gentamicin to determine whether a meaningful analysis of nutritional status can be used to identify patients with an expanded gentamicin volume of distribution (V) . Adults with gram-negative infections were selected sequentially from among hospitalized patients being treated with gentamicin . Serum gentamicin concentrations before and after an intravenous dose of gentamicin were determined by an enzyme-mediated immunoassay . Noncompartmental analysis was used to determine pharmacokinetic variables . Body composition was assessed by bioelectric impedance techniques . Of 26 patients, 10 were considered to be at high risk of malnutrition by meeting at least two of the following criteria: serum albumin concentration, less than or equal to 3.5 g/dL; actual weight, less than 80% of ideal weight; and ratio of exchangeable sodium to exchangeable potassium (Nae/Ke), greater than 1.22 . The non-high-risk group (n = 16) had a gentamicin V at steady state of 0.294 +/- 0.105 L/kg (mean +/- S.D.), which was significantly lower than that observed in the high-risk group (0.415 +/- 0.294 L/kg) . The accuracy of the high risk classification in identifying patients with an expanded gentamicin V was improved by revising the definition of the high-risk group . In the revised definition, patients were considered to be at high risk for malnutrition if they met at least two of the following criteria: serum albumin concentration, less than or equal to 3.1 g/dL; actual weight, less than 90% of ideal weight; and Nae/Ke' greater than 1.22.(ABSTRACT TRUNCATED AT 250 WORDS) Urology, 1990 Jan, 35(1), 54 - 6 Seminal antisperm antibodies and genitourinary infection; Micic S et al.; A total of 326 infertile men, 126 with genitourinary tract infection, and 200 without infection, were compared with 60 fertile men . Seminal plasma antisperm antibody with other clinical and microbiologic investigations were performed in all . Infertile men with genitourinary tract infection had antisperm antibody in 11.9 percent, infertile men without infection in 14.5 percent, and fertile men in 5 percent; these differences were not significant . The most common causes of these infections were gram-negative organisms and Chlamydia . Comparison of the levels of the titers of seminal plasma antisperm antibodies showed no significant difference between infertile and fertile men . This study shows no significant relationship between genitourinary tract infection and formation of the seminal antisperm antibody. Comp Biochem Physiol B, 1990, 97(3), 477 - 81 Membrane damage to Escherichia coli and bactericidal kinetics by the alternative complement pathway of channel catfish; Jenkins JA et al.; 1 . Increased permeability of cytoplasmic membranes in Escherichia coli was a consequence of alternative complement pathway (ACP) activity of serum of channel catfish, Ictalurus punctatus . Evidence was provided by beta-galactosidase activity extracellularly when E . coli was incubated with catfish serum . 2 . Lesions were detected on outer membranes of E . coli following exposure to catfish serum . 3 . Catfish ACP induced a temporal sequence of pre-killing and killing phases . 4 . Loss of cell viability, killing rate and cytoplasmic enzyme release increased with increasing serum concentrations . 5 . By incubating E . coli with sera treated to remove complement, both release of cytoplasmic enzyme and bactericidal activity were eliminated . 6 . Lethal activity associated with channel catfish ACP against Gram-negative bacteria was functionally comparable to that seen in mammalian and reptilian systems. Ann Med Interne (Paris), 1990, 141(6), 515 - 8 {Specificity and serum concentrations of tumor necrosis factor in septic shock}; Guidet B et al.; Several lines of evidence implicate tumor necrosis factor (TNF), a cytokine produced by monocytes-macrophages, in the systemic manifestations of shock induced by Gram-negative bacteria . Whether the increase of circulating TNF levels is specific to septic shock as compared to sepsis without shock or to non-septic shock is still unclear . Since TNF values recorded at the time of admission to the hospital vary widely, statistical analysis has not been possible . Therefore, we postulated that the evolution of a patient's TNF serum level as compared to his initial value may better distinguish the survivor from the non-survivor than a single initial determination . Using a radioimmunoassay, we measured the TNF concentrations in the sera of 7 patients with severe infections without shock, 16 patients with septic shock and 8 patients with non-septic shock . Blood samples were drawn within the first 12 hours after the onset of shock . Patients with cancer, HIV infection, or under steroid therapy were excluded . Repeated measurements were made during the first 3 days of septic shock in 10 patients . The circulating TNF level, determined upon admission, appears to be neither specific nor predictive of the outcome of septic shock . In contrast, persistently high levels of circulating TNF seem to be well correlated with a poor prognosis, since 5 out of 6 patients with elevated TNF values died of septic shock. Rev Mal Respir, 1990, 7(6), 497 - 503 {Respiratory manifestations of HIV infections in children}; de Blic J; Respiratory disorders during the course of infections in children who are HIV positive are frequent . These are interstitial pneumonia (IP) and bacterial pneumonia . The acute IP are most often infectious in particular opportunist infection, and are dominated by Pneumocystis carinii (PC, 82 to 86% of the infectious agents isolated) . This is often a presenting feature of an HIV infection and the radiological picture is very variable and the diagnosis rests on bronchoalveolar lavage which is readily achievable even in the infant . The immediate outcome is usually favourable and cotrimoxazole is efficacious and well tolerated . However pneumonia to PC points to a poor prognosis because they are always associated with a profound deficit or cellular immunity . All these criteria are against the likelihood of lymphoid PI which is, however, much more frequent in children than in adults . It is the chronic interstitial pneumonia with a "large grain" miliary radiological pattern and preserved cellular immunity and the absence of opportunistic infections in particular PC pneumonia . In addition, beyond the absence of PC, the bronchoalveolar lavage liquid shows a constant hyperlymphocytosis without polymorphonuclear leucocytes which is significantly different from that which is observed during the course of PC pneumonia . The long term prognosis of chronic PI, however, remains guarded with the possibility of seeing a worsening of the immune function . As for bacterial pneumonias they usually present as an acute lobar pneumonia with a banal organism but severe gram negative pneumonias are possible justifying a detailed systematic approach in certain cases. Life Sci, 1990, 47(21), 1933 - 9 Effect of a new synthetic free radical scavenger, 2-octadecyl ascorbic acid, on the mortality in mouse endotoxemia; Nonaka A et al.; Oxygen-derived free radicals have been implicated as mediators of cellular injury in several model systems . In order to clarify the role of oxygen radicals in endotoxemia, we measured the serial lipid peroxide changes resulting from systemic radical reactions using a newly developed colormetric method . To determine the effect of a free radical scavenger on mortality in endotoxemia, a new synthetic scavenger, 2-Octadecylascorbic acid (CV-3611), which overcome the detrimental properties (circulation half-life and cell penetration) of native SOD, was used in the model of mouse endotoxemia induced by the i.p . administration of E-coli endotoxin (10 mg/kg) . Serial LPO (Lipid Peroxide) changes revealed significant elevations from the basal level of 4.52 +/- 0.79 nmol/ml to 10.5 +/- 2.04 nmol/ml at 2h (P less than 0.05), 12.0 +/- 2.44 nmol/ml at 8h (P less than 0.05), 32.8 +/- 12.5 nmol/ml at 12h (P less than 0.05) and 13.6 +/- 2.40 nmol/ml at 24h (P less than 0.05) following i.p . administration of E-coli . The circulation half life of CV-3611 was checked by a reversed-phase HPLC after 10 mg/kg s.c . administration . The level of CV-3611 reached peak levels of 0.54 +/- 0.10 micrograms/ml at 1h and 0.52 +/- 0.20 micrograms/ml at 2h then gradually decreased to the level of 0.04 +/- 0.004 micrograms/ml at 6h and to a non-detectable level at 24h after s.c . administration . Increased survival was seen at 2 days (P less than 0.001) after E-coli endotoxin administration in the CV-3611 treated group compared to the control group . These results suggest that oxygen derived free radicals contribute to mortality in mouse endotoxemia and that antioxidants such as CV-3611 may provide a new therapeutic avenue by improving survival of patients with gram-negative bacterial sepsis. Crit Rev Microbiol, 1990, 17(5), 329 - 64 Family Spirosomaceae: gram-negative ring-forming aerobic bacteria; Raj HD et al.; The bacteria having a unique ring-like morphology first isolated from nasal mucus by Weibel in 1887 were classified as a new genus Spirosoma by Migula in 1894 . However, because these bacteria were not completely described for taxonomic purposes and their cultures were no longer available, the genus was deleted from the Bergey's Manual of Determinative Bacteriology, 6th edition, 1948 . Orskov (1928) created a new genus "Microcyclus" (a name that has been found to be illegitimate and replaced with Ancylobacter by Raj 1983) to describe these nonmotile vibroid bacteria that occasionally formed ring-like structures . Several similar isolates found in many countries during the last 60 years were readily identified with this genus on the basis of the characteristic morphology alone . For the first time, these fascinating bacteria were extensively reviewed by Raj in 1977 and again in 1981 . However, during the last decade, the systematics of these microcyclus bacteria has been reexamined and redefined . It has been shown that these Gram-negative ring-forming aerobic bacteria constitute a heterogeneous group of five genera: Ancylobacter, Cyclobacterium, Flectobacillus, Runella, and Spirosoma; the last four genera have been grouped into a family Spirosomaceace (reviving the old discarded name originally proposed by Migula 1894), thus separating them from the genus Ancylobacter which remains unaffiliated with any family yet (Bergey's Manual of Systematic Bacteriology, Vol . I, 9th ed., 1984) . Also, this article reviews the recent studies reported on the ecology, morphogenesis, metabolism, and physiology of the picturesque bacteria. Life Sci, 1990, 47(12), 1023 - 9 Pentoxifylline inhibits lipopolysaccharide-induced serum tumor necrosis factor and mortality; Noel P et al.; Tumor necrosis factor, a mononuclear phagocyte-derived peptide produced in response to lipopolysaccharide, has been shown to mediate certain aspects of septic shock and multiple organ failure resulting from gram-negative septicemia . In the present investigation, pretreatment of animals with pentoxifylline inhibited lipopolysaccharide-induced serum tumor necrosis factor in a dose-dependent fashion . Pentoxifylline prevented the sequestration of neutrophils seen in animals given intravenous lipopolysaccharide . Furthermore, pentoxifylline protected animals from the lethal effects of an intravenous challenge with lipopolysaccharide . These data indicate that pentoxifylline inhibits lipopolysaccharide-induced tumor necrosis factor and may be an effective agent in mitigating the lethal consequences of sepsis and other disease processes mediated by this cytokine. Am J Ind Med, 1990, 18(3), 279 - 84 Presence of endotoxins in different agricultural environments; Olenchock SA et al.; Gram-negative bacterial endotoxins are contaminants of dusts from agricultural products . They represent a potential health hazard for farmers working in many different processes . However, the occurrence of endotoxins has not been well characterized in the various farming operations . Therefore, two farming activities with potential for generating airborne endotoxins were studied: 1) chopping of baled corn stalks or straw for bedding in New York State, and 2) oat bin unloading in Alabama . Actual airborne endotoxin levels in dusts obtained during bedding chopper operations far exceeded {90 endotoxin units (EU)/m3} the level at which acute pulmonary function decrements occur in cotton dust-exposed individuals . Endotoxin contamination of laboratory-generated dust from an oat sample likewise exceeded these levels . This study documents the presence of potentially hazardous exposures to endotoxins in two common farm processes, which expands the knowledge of airborne endotoxin exposures on the farm. Intensive Care Med, 1990, 16(5), 307 - 11 Intestinal decontamination in a polyvalent ICU . A double-blind study; Godard J et al.; A double blind, placebo-controlled trial was performed to test the efficacy of prevention of nosocomial infections by selective digestive decontamination . Placebo or tobramycin (80 mg) and colistin (100 mg) was given four times daily via the gastric tube . Amphotericin B (500 mg/6 h) was administered to all patients . As our ICU is divided into two separate subunits, intestinal decontamination or placebo was administered alternatively to patients of the two subunits during two 3-month periods, separated by a 2-month period without prevention . The decontamination (n = 97) and placebo groups (n = 84) were similar with respect to age, sex, severity score and diagnostic categories on admission . Intestinal decontamination alone failed to significantly reduce the number of infected patients (26% vs 34.5%, p = 0.20), but was effective on ICU-acquired infections (0.33 vs 0.60, p = 0.02) especially gram-negative infection rates (0.17 vs 0.43, p = 0.01) . The onset of the first ICU-acquired infection was delayed (9 vs 13 days, p less than 0.001) and incidence of pneumonia (2 vs 13 cases, p less than 0.01) including bacterial pneumonia (0 vs 8 cases, p less than 0.01) was significantly decreased . However, mean ICU stay and mortality were not significantly modified by intestinal decontamination. Intensive Care Med, 1990, 16(5), 303 - 6 Gentamicin volume of distribution in critically ill septic patients; Triginer C et al.; Gentamicin intrapatient pharmacokinetics variations were studied in 40 critically ill medical patients, suffering gram-negative sepsis . These patients were studied in two phases throughout gentamicin treatment: firstly, on the second day of treatment, when aggressive fluid therapy was required, and secondly, five days later, when patients had achieved a more stable clinical condition . Pharmacokinetic parameters were determined using least squares linear regression analysis assuming a one-compartment model using the Sawchuk-Zaske method . The apparent volume of distribution (Vd) in the first phase of the study was 0.43 +/- 0.12 L/kg, while on the seventh day of treatment it was 0.29 +/- 0.17 L/kg (p less than 0.001) . Statistically significant differences were also observed for peak serum concentration (p less than 0.001), total dosage recommended (p less than 0.001) and half-life (p less than 0.05), whilst differences were not found for trough levels . From the analysis of the results obtained, we recommend increasing the initial dosage and monitoring plasma levels within the first days of therapy in critically ill patients treated with gentamicin, since important variations in aminoglycoside Vd related to disease, fluid balance and renal function, commonly occur in these patients. Prog Clin Biol Res, 1990, 349, 1 - 9 Regulation and role of phospholipases in host-bacteria interaction; Elsbach P et al.; Deacylating phospholipases play essential roles in numerous biological events, requiring tight control of hydrolytic activity . Most cells, unless stimulated or perturbed, exhibit little phospholipid turnover . Activation of phospholipases A (PLA) is usually triggered by membrane perturbing conditions or agents . Some activators indiscriminately activate any PLA, others are highly specific . Our studies concern an activator that is a potent bactericidal protein with membrane-perturbing properties, isolated from polymorphonuclear leukocytes (PMN), that is only cytotoxic for gram-negative bacteria and primarily responsible for the fate of several gram-negative bacterial species, ingested and killed by the PMN . It is this protein that activates the hydrolysis of the phospholipids of the killed bacteria (E . coli) by three PLA: 1) an E . coli PLA, the pldA gene product; 2) a PLA2 of PMN; 3) a soluble PLA2 in the extracellular fluid of an inflammatory exudate . However, this activator protein does not trigger the action of many other PLA2, all members of a highly conserved class of PLA . Our structural studies (including genetic engineering) of both responsive and non-responsive PLA2 have revealed that the amino acid composition and sequence of the NH2-terminal alpha-helix of the PLA2 molecule are major determinants of the ability of the PMN protein to activate a given PLA2 . Our results provide another demonstration that these important enzymes have diverged during evolution to perform different biological functions. Arkh Patol, 1990, 52(4), 9 - 16 {The morphofunctional characteristics of the microvascular endothelium and other structural elements of the lung in the development of gram-negative infection}; Barshtein IuA et al.; An early response (activation) of the microcirculatory bed endothelial cells precedes other changes in the lung in the course of development of gram-negative infection . This response is manifested at various degree in different endotheliocytes and taking into consideration the microenvironment (alveolar macrophages, stromal elements, etc) is one of the main mechanisms in the development of infectious process. Ann N Y Acad Sci, 1990, 587, 325 - 31 Metabolic responses to cachectin/TNF . A brief review; Tracey KJ et al.; Cachectin/tumor necrosis factor (TNF-alpha) is a macrophage-secreted cytokine initially found to be a lipoprotein lipase-suppressing serum factor in cachectic, parasite-infected animals . Cloning of the cDNA encoding the gene for cachectin enabled biosynthesis of recombinant human cachectin and proof that the protein is identical to TNF-alpha . Numerous biological activities have subsequently been attributed to this pluripotent cytokine . In addition to suppressing LPL, cachectin/TNF mediates decreased lipogenic enzyme synthesis in adipocytes, causing a state of "cellular cachexia" in vitro . Similarly, catabolic cellular energy responses are induced by cachectin/TNF in cultured skeletal muscle cells which exhibit accelerated glycogenolysis, enhanced lactate production, and increased expression of hexose transporters . Persistent cachectin/TNF production occurs in chronic infection and malignancy, and chronic exposure induces a cachexia syndrome characterized by anorexia, weight loss, and anemia . Acute systemic appearance of cachectin/TNF is capable of inducing a state of lethal shock, disseminated hemorrhagic necrosis, catabolic hormone release, and multiple organ injury . Inhibiting the toxic effects of cachectin/TNF with monoclonal anti-cachectin antibodies during overwhelming Gram-negative bacteremia confers protection against septic shock . In these studies, the unprotected controls succumbed within hours, but baboons immunized against cachectin/TNF did not develop the characteristic increases of IL-1, IL-6, or catabolic stress hormones and did not die, suggesting that cachectin/TNF is a pivotal, proximal factor in the humoral cascade mediating septic shock syndrome . Recent evidence indicates that when produced in lesser quantities, cachectin/TNF may participate in the degradative and reparative mechanisms of physiological tissue remodelling and homeostasis . Future studies of the immunological and metabolic effects of cachectin/TNF should lead to a better understanding of the pathogenesis of infection and inflammation. Adv Exp Med Biol, 1990, 256, 641 - 52 Lipid A precursors protect against endotoxin challenge; Proctor RA; These studies provide exciting prospects for the future treatment of gram-negative infections . The anti-endotoxin activity of lipid X, a monosaccharide precursor of lipid A, may be a prototypic compound for agents that can block the toxic effects of endotoxin that is being actively released . In contrast, monophosphoryl lipid A, a disaccharide derivative of lipid A, stimulates host defenses against infections and tumors . With further understanding of the mechanisms by which these compounds exert these effects, we can anticipate that new and more active compounds will be developed and that further activities of existing compounds will be found. Adv Exp Med Biol, 1990, 256, 233 - 45 Fluorescent detection of lipopolysaccharide interactions with model membranes; Jacobs DM et al.; The critical importance of the lipid A moiety of LPS in resistance and pathogenesis in gram negative infections has led to the assumption that LPS interaction with target cells is due to hydrophobic interaction with plasma membranes . However, work from several laboratories, including our own, is consistent with the presence of a cell membrane structure with characteristics of a "receptor" . We have proposed a two-step model for LPS-membrane interaction which resolves the two views, and have developed a model system to control the first step (binding to membrane protein) and study the second step (intercalation into lipid bilayer) . We examined the interaction of LPS with small unilamellar phosphatidylcholine vesicles labeled in the hydrophobic portion of the bilayer with the fluorescent probe diphenylhexatrine (DPH) and detected changes in the physical properties of the bilayer by measuring DPH fluorescence anisotropy (delta r) . We have found that purified, phenol-extracted S . typhimurium LPS interacts with the bilayer as measured by an increase in delta r and conclude that the LPS aggregate coalesced with the lipid bilayer . The greatest change in delta r was achieved with lipid A, Ra-Re glycolipids and diphosphoryl lipid A . Monophosphoryl lipid A and lipid X were less effective . Preparations of wild-type LPS fractionated according to the length of the O-antigen side chain and unfractionated LPS had least effect on delta r . Thus other factors such as serum components or membrane proteins may be necessary to enhance the interaction of LPS with target cells. Scand J Infect Dis, 1990, 22(1), 79 - 86 Continuous removal of leucocytes during early gram-negative septicemia reduces plasma levels of endotoxin and improves cardiac performance; Rokke O et al.; The effect of filter leucapheresis as treatment of gram-negative septicemia was studied in an experimental porcine model . In 7 matched pairs of animals, one of the animal groups received filter leucapheresis starting 2 h after the induction of septicemia . The other animal group acted as a non-treated control . A calculated mean number of 15 x 10(9) white blood cells were removed . Animals submitted to leucapheresis showed lower levels of circulating endotoxin, diminished degree of granulocyte activation and higher cardiac output when compared to the non-treated controls . The increased cardiovascular performance indicated a beneficial effect of selective leucafiltration during early gram-negative septicemia . Lower levels of circulating endotoxin and less active PMNs might contribute to these changes. Mol Microbiol, 1990 Jan, 4(1), 123 - 8 Isolation of the Actinobacillus pleuropneumoniae haemolysin gene and the activation and secretion of the prohaemolysin by the HlyC, HlyB and HlyD proteins of Escherichia coli; Gygi D et al.; The gene encoding the c . 105 kD secreted haemolysin protein of the porcine pathogen Actinobacillus pleuropneumoniae serotype 1 has been isolated by screening a lambda gt11 expression library in Escherichia coli with antiserum raised against the wild-type protein . A derivative recombinant DNA pJFF702 expressed the hlylA haemolysin gene from the pUC19 lac promoter but the resulting haemolysin I protein remained within the E . coli cell and was haemolytically inactive . Export of the intracellular A . pleuropneumoniae prohaemolysin out into the medium was achieved by the presence in trans of the E . coli haemolysin secretion genes hlyB and hlyD, and high levels of intracellular haemolytic activity were attained similarly by the E . coli post-translational haemolysin activator gene, hlyC . Southern hybridization of A . pleuropneumoniae parental DNA nevertheless indicated only a low degree of nucleotide sequence identity to the haemolysin structural and secretion genes hlyA and hlyB of E . coli . The data show that despite substantial nucleotide sequence divergence the A . pleuropneumoniae serotype 1 haemolysin determinant is closely related to that which is dispersed throughout other Gram-negative human and animal pathogens. J Basic Microbiol, 1990, 30(4), 279 - 87 DNA probes for studying streptothricin resistance evolution in enteric bacteria; Tietze E et al.; Probes for the detection of streptothricin resistance genes have been derived from recombinant plasmids . These include the streptothricin resistance gene probe sat 1/2 derived from Tn 1826 and specific for both the sat-1 determinant of Tn 1825 and the sat-2 determinant of Tn 1826, and the probe sat D derived from and specific for the sat-1 determinant of transposon Tn 1825 . A third streptothricin resistance gene probe, sat 3, represents the streptothricin resistance determinant sat-3 of the IncQ R plasmid pIE639 . Hybridization studies did not reveal any sequence homology between sat-3 and the transposon-localized sat-1 and sat-2 determinants . Moreover, non of the different sat-determinants isolated from plasmids of gram negative bacteria hybridized with the analogous resistance determinant of Streptomyces noursei, which had been cloned and named nat by Krugel et al . (Gene, 1988, 62, 209-214) . The sat 1/2 probe in combination with the sat D probe proved to be suitable for the identification and the differentiation of sat-1 and sat-2 determinants in different genetic environments . Streptothricin resistance genes related to those present on transposons Tn 1825 and Tn 1826 have been detected by hybridization with the probe sat 1/2 on plasmids isolated a long time ago before the application of streptothricins . The sat-3 determinant appears to be exclusively associated with the IncQ plasmid pIE639. Lymphokine Res, 1990 Summer, 9(2), 125 - 35 Cholera and pertussis exotoxins protect mice against the lethal Schwartzman reaction and antagonize the effects of lipopolysaccharide on second messenger systems; Cabeza-Arvelaiz Y et al.; Pertussis toxin, and also cholera toxin are capable of inhibiting the effects of LPS in the elicitation of the generalized Schwartzman reaction . This is a potentially lethal generalized thrombo-haemorrhagic hypersensitivity and inflammatory-type response that occurs after two consecutive injections of LPS . The two exotoxins furnish significant protection against the lethal outcome of this reaction . It is known that the acute haematological and haemodynamic changes are accompanied by alterations in the levels of various endogenous mediators: glucocorticoid hormones, prostaglandins, arachidonic acid metabolites, cytokines and proteases . In vitro effects of LPS on murine leukocyte cell lines can be antagonized by pertussis toxin, implicating a Gi-like regulatory protein in the mediation of these effects . Experiments designed to study the involvement of particular second messenger systems (cAMP and phosphatidylinositol) used by LPS in vivo, revealed that the protective effects conferred by these exotoxins are associated with the antagonization of alterations caused by LPS . No correlation was found between the levels of IL-6 and the mortality rate in this experimental mouse model . The results indicate that G proteins play a role in the generation of the Schwartzman reaction and open a new approach for pharmacological intervention in endotoxemia and in clinical settings with Gram-negative sepsis. J Allergy Clin Immunol, 1990 Jan, 85(1 Pt 1), 45 - 51 The effects of intravenous endotoxin on various host-effector molecules; Casale TB et al.; To understand better the pathogenesis of gram-negative infections, we administered a single intravenous injection of Escherichia coli RE-2 endotoxin (4 ng/kg) to five normal volunteers and periodically measured various host-effector molecules in blood . All subjects had a significant (p less than 0.05) increase in temperature (maximum of 1.3 +/- 0.1 degrees C at 4 hours) and white blood cell count (maximum of 7700 +/- 1800 cells per cubic millimeters at 8 hours) . Thymocyte costimulatory activity in blood was markedly increased 2 hours after endotoxin administration but returned to baseline at 4 hours . Blood cortisol and beta-endorphin levels paralleled each other, peaking 4 hours after endotoxin (mean increases of 21 +/- 14 micrograms/dl and p less than 0.01; 18.6 +/- 5.9 pg/ml and p less than 0.05, respectively) and returning to baseline at 8 hours . Blood histamine levels did not significantly change at any time point after endotoxin administration . Since thymocyte costimulatory activity was the first of the measured parameters to achieve peak blood levels, we suggest that endotoxin-induced increases in various cytokines may contribute to the increases in the other measured parameters and thereby play a significant role in the pathogenesis of gram-negative infections.