Gram negative

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.

Arch Invest Med (Mex), 1990, 21 Suppl 1, 175 - 81
{Immunochemical differences in the surface polysaccharides obtained from Entamoeba histolytica strain HM1:IMSS and its virulent (C-A) and non-virulent (L-6) clones}; Isibasi A et al.; There probably exist many different surface molecular polysaccharides with different compositions of their constituent sugars, in the different zymodemes . This fact might have a repercussion upon the resistance to the lysis of the complement . Seeking to prove this hypothesis, the ideal would be to obtain polysaccharide molecules of type LPFG of pathogenic and non-pathogenic zymodemes, demonstrating that there is a difference in the chemical composition of their polysaccharides . If this were true, a serologic classification, of the different zymodemes, would be possible in a fashion similar to gram-negative bacteria . Our results suggest the existence of structural differences between the polysaccharides of a virulent clone (C-A) and a non-virulent one (L-6).

Arq Cent Estud Curso Odontol, 1990 Jan-Dec, 27(1-2), 9 - 15
{Denture stomatitis--presence of bacteria and yeasts}; Jorge AE et al.; The presence of yeasts and bacteria was studied in 26 patients with denture stomatites, and the results compared with the data of the normal mucosa foi edentoulous patients, who used or not upper dentures . The use of dentures caused an increase in the amount of yeasts, and there was a correlation with the severity of the stomatitis . Gram positives cocci and bacilus predominated in all studied groups, but in cases of stomatitis there was an increase in the amount of Gram negative cocci and filamentous . These results suggest that besides yeasts, modifications of the bacteria flora can be relevant for the development of denture stomatitis.

Lung, 1990, 168 Suppl, 707 - 15
Biological activity of bacterial surface components: bacterial extracts and defined bacterial cell wall components as immunomodulators; Bessler WG et al.; Bacterial extracts obtained from pathogenic strains occurring in lung infections (Broncho Vaxom) or urogenital infections (Urovaxom) as well as defined surface components of Gram-negative bacteria purified from bacteria or obtained by chemical synthesis were tested for their immunomodulatory properties in a murine system . The bacterial extracts were able to act as immunogens inducing an antigen-specific response . Both the bacterial extracts and the purified bacterial cell wall components constituted polyclonal activators of murine splenic B cells, as demonstrated by proliferation assays measuring the incorporation of {3H}thymidine into DNA . They were also able to act as immunoadjuvants increasing the SRBC and the BSA-TNP specific immune response, and could induce tumor cytotoxicity in bone marrow-derived macrophages . The results show that bacterial extracts and defined bacterial surface components constitute immunogens as well as immunomodulators in vitro and in vivo.

Am J Physiol, 1990 Jan, 258(1 Pt 1), E57 - 64
Interleukin 6 stimulates hepatic glucose release from prelabeled glycogen pools; Ritchie DG; Cytokines, derived from a wide variety of cell types, are now believed to initiate many of the physiological responses accompanying the inflammatory phase that follows either Gram-negative septicemia or thermal injury . Because hypoglycemia (after endotoxic challenge) and hyperglycemia (after thermal injury) represent well-characterized responses to these injuries, we sought to determine whether hepatic glycogen metabolism could be altered by specific cytokines . Cultured adult rat hepatocytes were prelabeled with {14C}glucose for 24 h, a procedure that resulted in the labeling of hepatic glycogen pools that subsequently could be depleted (with concomitant {14C}glucose release) by either glucagon or norepinephrine . After the addition of a highly concentrated human monocyte-conditioned medium (MCM) or various cytokines to these prelabeled cells, {14C}glucose release was stimulated by MCM and recombinant human interleukin 6 (IL-6) but was not stimulated by other cytokines tested . Furthermore, only antisera to IL-6 were capable of reducing the glucose-releasing factor activity found in MCM . These data therefore suggest a novel glucoregulatory role for IL-6.

Int Rev Immunol, 1990, 6(4), 207 - 21
Bacterial lipopolysaccharides: structure, metabolism and mechanisms of action; Freudenberg MA et al.; Endotoxins (lipopolysaccharides, LPS) are biologically active substances present in the outer membrane of gram-negative bacteria . They induce a spectrum of biological effects which may be harmful or beneficiary for the host . Lipid A is the biologically active part of the LPS molecule . This was demonstrated using soluble forms of lipid A and more recently confirmed further by employing synthetic lipid A . LPS administered into experimental animals circulates as LPS/HDL complex and is cleared from the blood mainly into the liver and spleen . In the liver LPS undergoes partial deacylation however without a loss of toxic activity . Its excretion is effected mainly via the bile into the gut . The lethal toxicity and tolerance inducing properties of LPS are mediated by macrophages through tumor necrosis factor alpha (TNF alpha), which is probably the most important endogenous mediator of the lethal effects of LPS . The lethal toxicity of LPS may be completely inhibited by anti-TNF alpha antibodies.

Lymphokine Res, 1990 Winter, 9(4), 449 - 63
Differential release of TNF-alpha, IL 1, and PGE2 by human blood monocytes subsequent to interaction with different bacterial derived agents; Fidler IJ et al.; The purpose of these studies was to determine whether the tumoricidal phenotype of human blood monocytes would be affected by different activation signals . Human monocytes obtained by elutriation of buffy coats were cultured in vitro in medium containing LPS, muramyltripeptide phosphatidylethanolamine (MTP-PE), or a lipopeptide analogue of gram-negative bacteria cell wall . These immunomodulators were added to monocytes in the presence or absence of IFN-gamma . Incubation with LPS, lipopeptide, and MTP-PE rendered the monocyte cytotoxic against allogeneic melanoma cells . Monocytes treated with LPS and lipopeptide (in the absence of IFN-gamma) secreted IL 1, TNF, and PGE2 . In contrast, monocytes incubated with MTP-PE (in the absence of IFN-gamma) secreted only TNF . When the monocytes were coincubated with IFN-gamma (human but not mouse) and the immunomodulators, IL 1, TNF, and PGE2 were secreted at all test groups . These data show that some immunomodulators can regulate the release of TNF independently of IL 1 and that not all "activated tumoricidal macrophages" share identical phenotypes.

Am J Nephrol, 1990, 10(5), 397 - 403
Outbreak of gram-negative bacteremia and pyrogenic reactions in a hemodialysis center; Beck-Sague CM et al.; During the period from April 4, 1988, to April 20, 1988, nine pyrogenic reactions and five gram-negative bacteremias occurred in 11 patients undergoing dialysis . All pyrogenic reactions and gram-negative bacteremias occurred among patients in whom a reprocessed dialyzer was used . The rate of pyrogenic reactions or bacteremias per 100 sessions using a reprocessed dialyzer was higher than in sessions during which a new dialyzer was used (4.5 vs . 0; p = 0.03) . Dialyzers were manually reprocessed with 2.5% Renalin germicide . The Renalin concentrations varied widely in 12 dialyzers stored after manual reprocessing during the epidemic period (0.9-4.2%); the median endotoxin concentrations varied from 0 to 246 ng/ml and were higher in dialyzers with Renalin concentrations less than or equal to 1.0% than in dialyzers with higher concentrations (p = 0.01) . Experiments using a dilution technique described by a technician resulted in Renalin concentrations ranging from 1.4% at the surface to 3.5% at the bottom of the preparation container . These findings suggest that failure to adequately admix Renalin during dilution may be associated with low levels of disinfectant, high levels of bacteria and endotoxins in dialyzers, and outbreaks of pyrogenic reactions and gram-negative bacteremias in dialysis patients.

Microbiol Rev, 1989 Dec, 53(4), 491 - 516
Replication of plasmids in gram-negative bacteria; Kues U et al.; Replication of plasmid deoxyribonucleic acid (DNA) is dependent on three stages: initiation, elongation, and termination . The first stage, initiation, depends on plasmid-encoded properties such as the replication origin and, in most cases, the replication initiation protein (Rep protein) . In recent years the understanding of initiation and regulation of plasmid replication in Escherichia coli has increased considerably, but it is only for the ColE1-type plasmids that significant biochemical data about the initial priming reaction of DNA synthesis exist . Detailed models have been developed for the initiation and regulation of ColE1 replication . For other plasmids, such as pSC101, some hypotheses for priming mechanisms and replication initiation are presented . These hypotheses are based on experimental evidence and speculative comparisons with other systems, e.g., the chromosomal origin of E . coli . In most cases, knowledge concerning plasmid replication is limited to regulation mechanisms . These mechanisms coordinate plasmid replication to the host cell cycle, and they also seem to determine the host range of a plasmid . Most plasmids studied exhibit a narrow host range, limited to E . coli and related bacteria . In contrast, some others, such as the IncP plasmid RK2 and the IncQ plasmid RSF1010, are able to replicate in nearly all gram-negative bacteria . This broad host range may depend on the correct expression of the essential rep genes, which may be mediated by a complex regulatory mechanism (RK2) or by the use of different promoters (RSF1010) . Alternatively or additionally, owing to the structure of their origin and/or to different forms of their replication initiation proteins, broad-host-range plasmids may adapt better to the host enzymes that participate in initiation . Furthermore, a broad host range can result when replication initiation is independent of host proteins, as is found in the priming reaction of RSF1010.

J Gen Microbiol, 1989 Dec, 135 ( Pt 12), 3253 - 62
Purification and partial characterization of a major outer-membrane protein of Fusobacterium nucleatum; Bakken V et al.; The major outer-membrane proteins of 40-41 kDa were identified by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in Fusobacterium nucleatum strains ATCC 10953, ATCC 25586, F3, F6 and Fev1 . The proteins were purified by preparative gel electrophoresis . Their behaviour in gel filtration and gel electrophoresis, their sensitivity to proteolytic enzymes, and their amino acid composition were investigated . The purified proteins were partly sequenced from the N-terminal end . A 36.5 kDa portion was protected against extrinsic proteolytic (trypsin, chymotrypsin or pronase) digestion of whole cells . This polypeptide was isolated and partially sequenced from the N-terminal end . From these data and data from extrinsic iodination it was concluded that the N-terminal end of the protein is probably exposed on the surface of the cell . A database search revealed amino acid sequence similarity in an Ala-Pro-rich region of outer-membrane protein A (OmpA) in other Gram-negative bacteria.

Biochem Int, 1989 Dec, 19(6), 1223 - 9
Purification and properties of RNA polymerase from acidophilic heterotroph SJH; Beebee TJ; RNA polymerase was isolated from the acidophilic heterotrophic bacterium SJH and purified to homogeneity . Subunit structure (beta', beta, theta, alpha 2) resembled that of other gram-negative bacteria such as Escherichia coli . pH optimum of the enzyme was around 8.0, although the bacterium grew only below pH 6.0 and optimally between 3.0-4.0 . SJH and E . coli RNA polymerases showed maximal activities on homologous high molecular-weight DNA templates.

Zentralbl Bakteriol, 1989 Dec, 272(2), 225 - 30
Synergism between ciprofloxacin and fosfomycin against gram-negative bacteria in vitro; Vogt K et al.; A combination of ciprofloxacin and fosfomycin was evaluated for synergism employing time-kill experiments . A total of 59 clinical isolates of gram-negative bacteria was examined . All organisms were moderately sensitive or resistant to ciprofloxacin; 72.9% also showed moderate sensitivity or resistance to fosfomycin . After 24 h, synergism was seen in 22% of the strains, i.e . less than reported by other authors . There was no correlation between synergistic action and sensitivity to ciprofloxacin and fosfomycin, respectively.

Arch Fr Pediatr, 1989 Dec, 46(10), 741 - 2
{Nephrotic syndrome and B leukemia}; Pacquement H et al.; A boy, aged 14 1/2 years, presented with Burkitt leukemia . His renal status was normal before treatment . Chemotherapy (SFOP LMB 86 protocol) was begun Oct . 9, 1986 . After the first 2 courses of chemotherapy, the patient had Gram negative sepsis treated with cefotaxime, netilmycine, Vancomycin and ornidazole . During sepsis, nephrotic syndrome developed (albumin 25 g/l, non selective proteinuria 15 g/24 h), with moderately high blood pressure, functional renal failure (creatinine 141 mumols/l, U/P urea = 20), polyuria and tubular damage . Kidney ultrasonography was normal . Needle biopsy showed minimal glomerular lesions, acute tubular lesions, and no deposits in immunofluorescence . The nephrotic syndrome disappeared within 3 weeks, with treatment of leukemia . He is at present in complete remission with a follow-up of 25 months.

Carbohydr Res, 1989 Dec 1, 194, 209 - 21
Studies of the synthesis of sugar phosphonates related to 3-deoxy-D-manno-2-octulosonic acid (Kdo); Molin H et al.; In attempting to synthesize the analogue of beta-Kdo (2R)-2-carboxy-6-(1',2'-dihydroxyethyl)-4,5-dihydroxy-D-manno-1,2 lambda 5-oxaphosphorinan-2-one (6) as an inhibitor of the enzyme CMP-Kdo synthetase, which is involved in the biosynthesis of the lipopolysaccharide component of the outer membrane of Gram-negative bacteria, (2R)-6-(1',2'-dihydroxyethyl)-2-ethoxy-3,4,5-trihydroxy-4,5:1', 2'-di-O-isopropylidene-D-glycero-D-talo-1,2 lambda 5-oxaphosphorinan-2-one (8) was converted into (2S)-6-(1',2'-dihydroxyethyl)-4,5-dihydroxy-4,5:1',2'-di-O-isop rop ylidene-2-vinyl-D-manno-1,2 lambda 5-oxaphosphorinan-2-one (16), but alkene cleavage to give the target carboxyphosphonate failed . Reduction-oxidation-Arbuzov reaction on the intermediate (2R)-6-(1',2'-dihydroxyethyl)-2-ethoxy-4,5-dihydroxy-4,5:1', 2'-di-O-isopropylidene-D-manno-1,2 lambda 5-oxaphosphorinan-2-one (11) gave the 2S isomer of the protected target compound, but removal of the protecting groups gave the acyclic product dilithium (D-manno-2,3,4,5,6-pentahydroxyhexyl)phosphinatoformate (24) . N.m.r . studies of the intermediates allowed assignment of stereochemistry at P for all compounds via 2JP,H coupling constants.

Scand J Work Environ Health, 1989 Dec, 15(6), 430 - 5
Effects of different extraction protocols on endotoxin analyses of airborne grain dusts; Olenchock SA et al.; The detection of gram-negative bacterial endotoxins in occupational dusts, specifically those from agricultural environments, is of increasing importance in research on occupational lung disease . In this study, the quantitative chromogenic Limulus amebocyte lysate test for the detection of endotoxins in airborne dusts from spring wheat and oats was examined . Different extraction fluids were tested, as were the effects of time on extraction and of repeated freeze-thaw cycles on the extracts . The data suggest that the chromogenic method can be used effectively in the analysis of environmental dusts or their frozen extracts for endotoxin quantitation . Water appears to be the preferable extraction medium, and the length of extraction time may affect the results.

Neurosurgery, 1989 Dec, 25(6), 923 - 6
Causes of infections in penetrating head wounds in the Iran-Iraq War; Aarabi B; Factors seemingly influential in postoperative central nervous system infections were evaluated in 379 patients who sustained missile wounds to the head during the Iran-Iraq War . The mean wound age was 49 hours . The site of injury and presence of retained bone and/or shell fragments did not have a significant effect on infection rate . Thirty-three of 379 patients developed cerebrospinal fluid (CSF) fistulas of which 12 (36%) were associated with infection . The infection rate was 6 of 346 (1.7%) in the absence of CSF leaks . There was a statistically significant association between CSF fistula and infection . All 6 positive cultures in patients with CSF fistulas were gram-negative . Thirty-two patients died during the study, a mortality rate of 8.4% . The mortality rate from infection was 1.8% . Two hundred seventy-seven of 347 patients were followed for a mean of 22 +/- 17 months during which time 2 patients were readmitted with central nervous system infections, both due to CSF leakage . CSF fistulas were the main predisposing factor in postdebridement central nervous system infections in this study.

Arch Surg, 1989 Dec, 124(12), 1416 - 20; discussion 1420-1
Kupffer cell cytotoxicity to hepatocytes in coculture requires L-arginine; Billiar TR et al.; Activated macrophages convert L-arginine to citrulline and unstable nitrogen oxides that have cytotoxic properties . We recently have shown that the inhibition of protein synthesis in Kupffer cell (KC):hepatocyte (HC) coculture, following exposure to gram-negative bacterial endotoxin (lipopolysaccharide), is due to the metabolism of L-arginine by this cytotoxic pathway . Although this finding supports a role for activated KCs and the L-arginine-dependent mechanism in the HC dysfunction seen in sepsis, it and previous studies have failed to demonstrate direct damage to HCs by adjacent KCs . The current study was undertaken to determine if KCs exposed to lipopolysaccharide could directly damage HCs and, if so, whether the damage was dependent on the metabolism of L-arginine . By using the release of aspartate aminotransferase as a marker of HC damage, it was found that a significant aspartate aminotransferase release by KC:HC cocultures in response to lipopolysaccharide occurred only if L-arginine was present . In addition, requirements for significant aspartate aminotransferase release included KC:HC ratios of 7.5:1 or greater and L-arginine concentrations of 1 mmol or more . Although the KC-induced damage was mild, these results show that in vitro HC damage in KC:HC coculture does require the metabolism of L-arginine and supports a hypothesis that toxic L-arginine metabolites may contribute to liver cell damage in patients with sepsis.

J Infect Dis, 1989 Dec, 160(6), 1005 - 11
Cerebrospinal fluid endotoxin levels in children with H . influenzae meningitis before and after administration of intravenous ceftriaxone; Arditi M et al.; Total, cell-free, and cell-bound endotoxin and bacterial density were measured in cerebrospinal fluid (CSF) of 22 children with Hemophilus influenzae meningitis . Also the effect of ceftriaxone on CSF endotoxin levels was investigated in eight patients by reexamining their CSF 2-6 h after the initial dose . Initial CSF bacterial density correlated with initial CSF endotoxin levels (P less than .001) . Ceftriaxone induced a marked increase of free endotoxin in CSF, from an initial (mean +/- SE) 0.75 +/- 0.21 to 1.29 +/- 0.23 log10 ng/ml (P less than .01) . This increase correlated positively with the number of bacteria killed in the CSF (P less than .01) . The increase in free endotoxin was associated with an increase in mean CSF lactate levels from 8.5 to 9.7 units/l (P less than .05) and mean lactate dehydrogenase levels from 102 to 180 mmol/l (P less than .02) and a decrease in mean CSF glucose from 1.17 to 0.46 mmol/l (P less than .05) . Initial CSF total endotoxin concentrations correlated both with the Herson-Todd clinical severity score (P less than .001) and with the number of febrile hospital days (P less than .001) . These findings suggest that highly bactericidal agents initially lead to release of free endotoxin from gram-negative organisms into CSF, with associated enhanced inflammatory response by the host.

Biofactors, 1989 Dec, 2(2), 87 - 94
PQQ and quinoprotein research--the first decade; Duine JA; On the occasion of the first international symposium on pyrroloquinoline quinone (PQQ) and quinoproteins (Delft, September 1988), a review of this novel field in enzymology is presented . Quinoproteins (PQQ-containing enzymes) are widespread, from bacteria to mammalian organisms (including man), and occur in several classes of enzymes . Indications already exist that PQQ is a versatile cofactor, involved not only in oxidation but also in hydroxylation, transamination, decarboxylation and hydration reactions . The current list of quinoproteins shows that it was overlooked in several well-studied enzymes where the presence of a common cofactor had already been established . Up until now, all eukaryotic quinoproteins have covalently bound PQQ (or perhaps pro-PQQ), while free PQQ occurs exclusively in a number of (bacterial) dehydrogenases and in the culture fluid of certain Gram-negative bacteria . Biosynthesis of free PQQ in methylotrophic bacteria starts with tyrosine and glutamic acid as precursors while intermediates in the route have not been detected and the presence of free PQQ is not required for synthesis of the covalently bound form of the cofactor in glutamic acid decarboxylase from Escherichia coli . Therefore, the assembly of covalently bound cofactor might occur in situ, i.e . in the quinoproteins themselves . If the latter also applies to mammalian quinoproteins, this implies that PQQ is not a vitamin . On the other hand, positive effects have been reported upon administration of PQQ to test animals . Methods suited to detach and to detect PQQ with a derivatized o-quinone moiety may answer questions on the uptake and processing of the compound.

J Biol Response Mod, 1989 Dec, 8(6), 625 - 43
T cell and interferon-gamma involvement in the adjuvant action of a detoxified endotoxin; Tomai MA et al.; The adjuvant activities of a detoxified derivative of endotoxic lipopolysaccharides, isolated from the outer membrane of gram-negative bacteria, were evaluated in aging mice . This monophosphoryl lipid A (MPL) (Ribi) was capable of enhancing antibody production in vitro in splenic cultures from 2-3-month-old male Balb/c mice as well as cultures from 22-24-month-old Balb/c mice . Separation of spleen cells from MPL and phosphate-buffered saline-injected mice into adherent and nonadherent populations and subsequent mixing of populations and culture with antigen implicated an adherent cell as being involved in the enhancement of antibody formation induced by MPL . However, separation of normal spleen cells into purified populations of adherent cells, T-lymphocytes, and B-lymphocytes, followed by in vitro stimulation of the individual populations with MPL and subsequent transfer into cultures of normal spleen cells, revealed only the T cell as capable of transferring the enhancement of antibody formation . In addition, culture filtrates from MPL-stimulated T cells were able to enhance antibody production by spleen cell cultures from aging mice twofold above that of filtrates from unstimulated T cells . The enhancement of antibody formation induced by such filtrates and also by MPL in spleen cell cultures from young and aging mice was inhibited by a monoclonal antibody (MAb) to recombinant interferon-gamma (rIFN-gamma) as well as antiserum against IFN-alpha, -beta, and -gamma, but not by an antiserum to IFN-alpha/beta . Enhancement of antibody formation correlated well with an increase in interleukin-1 (IL-1) but not with an increase in IL-2 production . Addition of anti-asialo-GM1 MAb plus complement to the effective spleen populations did not diminish the adjuvant action.

J Bacteriol, 1989 Dec, 171(12), 6521 - 5
Nucleotide transport in Rhodobacter capsulatus; Carmeli C et al.; Cytoplasmic membrane vesicles isolated from the gram-negative photosynthetic bacterium Rhodobacter capsulatus catalyzed the transport of nucleotides . No transport occurred in the intact bacteria unless they were pretreated with EDTA . The transport rate was measured by incorporation of radioactive phosphate into externally added ADP or by incorporation of nonradioactive phosphate into added labeled ADP . The catalytic activities which utilized the added ADP were photosynthetic ATP synthesis, Pi-ADP exchange, and adenylate kinase . These activities were shown to occur on the cytoplasmic side of the internal membrane . The products were found in the outer medium . The rate of nucleotide transport across the membranes was comparable to the rate of photophosphorylation . These results indicated that nucleotides can be transported across the cytoplasmic membrane but not across the outer membrane of the native R . capsulatus cell . Therefore, by analogy to the mitochondrial ATP-ADP translocator, the exchange might function as an energy transfer system to the periplasm of these bacteria.

J Clin Microbiol, 1989 Dec, 27(12), 2881 - 3
Identification of Moraxella bovis by using a monoclonal antibody to a lipopolysaccharide epitope; Wannemuehler Y et al.; A monoclonal antibody to the lipopolysaccharide of Moraxella bovis is described . In an indirect fluorescent-antibody test, this monoclonal antibody reacted with 39 of 39 strains of M . bovis tested and did not react with 26 nonfermenting gram-negative coccobacilli other than M . bovis . When used in an indirect fluorescent-antibody test, it proved useful for rapid and easy identification of M . bovis.

Gene, 1989 Nov 30, 83(2), 371 - 5
Bacteriophage T7 RNA polymerase-controlled specific gene expression in Pseudomonas; Davison J et al.; The rifampicin (Rif)-resistant RNA polymerase of phage T7 has proved invaluable for the exclusive over-expression, in Escherichia coli, of genes cloned downstream from the T7 phi 10 promoter {Tabor and Richardson, Proc . Natl . Acad . Sci . USA 82 (1985) 1074-1078} . Here, we demonstrate that the system can be extended to Gram-negative bacteria other than E . coli, by the use of compatible wide host range plasmids . As an example, the Rif-resistant in vivo synthesis and specific radiolabelling of E . coli galactokinase in Pseudomonas ATCC19151, is demonstrated . The incidental observation that 30 min after treatment with Rif, two polypeptides continue to be synthesized in plasmid-free Pseudomonas ATCC19151, indicates that these proteins are produced by very stable mRNA species.

Am J Med, 1989 Nov 30, 87(5A), 248S - 249S
Ciprofloxacin in the treatment of gram-negative bacillary meningitis; Schonwald S et al.; Gram-negative bacillary meningitis (typically caused by E . coli, K . pneumoniae, P . aeruginosa, A . calcoaceticus, and others) has been associated with head trauma, neurosurgical operations, pathologic anastomosis with nasal cavity, and CSF rhinorrhea, as well as with impaired host defenses . Intravenous ciprofloxacin was given to 20 patients with gram-negative bacillary meningitis . The dose of ciprofloxacin was 200 mg by intravenous infusion, over 30 minutes, every 12 hours for 10 days . Two patients also received intravenous cefotaxime and penicillin G . All patients were monitored daily . Of 20 patients, 18 had cure and therapy failed in two . Because relatively low concentrations of ciprofloxacin are achieved in the CSF, the drug should be used in the treatment of gram-negative bacillary meningitis only if the MICs of the causative pathogen and the ciprofloxacin concentration in CSF can be followed . A randomized, controlled study is needed to compare the efficacy of intravenous ciprofloxacin with standard antibiotic therapy in the treatment of this disease.

Cell, 1989 Nov 17, 59(4), 657 - 65
The molecular mechanism of phase variation of H . influenzae lipopolysaccharide; Weiser JN et al.; Multiple carbohydrate structures on the outer-membrane lipopolysaccharide (LPS) of the gram-negative pathogen H . influenzae undergo high frequency, reversible loss, indicative of phase variation . Characterization of a genetic locus, lic-1, responsible for expression of two LPS epitopes displaying phase variation, showed it to comprise four genes . The first gene mediates phase variation . At its 5' end, within the open reading frame, are a variable number of tandem repeats of the tetramer CAAT . By shifting upstream initiation codons in or out of frame, these 4 bp units create a translational switch . The phenotype of organisms corresponds to the number of 4 bp units . Phase variation between three levels of expression ( +, +, and -) of lic-1-derived epitopes is caused by differences in the three phases of translation of the 5' terminus of this gene . Phase variation also allows for selection of organisms displaying certain LPS epitopes in vivo.

J Exp Med, 1989 Nov 1, 170(5), 1627 - 33
Antibodies to cachectin/tumor necrosis factor reduce interleukin 1 beta and interleukin 6 appearance during lethal bacteremia; Fong Y et al.; Cytokines secreted in response to invading micro-organisms are important mediators of detrimental hemodynamic and metabolic changes in the host . To test whether cachectin/TNF plays a role in triggering release of other cytokines in the setting of infection, anesthetized baboons were passively immunized against systemic cachectin/TNF before infusion of a LD100 dose of live Escherichia coli . Bacteremia led to significant increases in circulating levels of cachectin/TNF, IL-1 beta, and IL-6 . Although bacterial endotoxin/lipopolysaccharide is a potent stimulus for the synthesis and release of IL-1 and IL-6 in vitro, specific neutralization of cachectin/TNF in vivo with mAb pretreatment significantly attenuated both the IL-1 beta and the IL-6 responses despite fulminant overwhelming bacteremia . These data suggest that cachectin/TNF is essential for the initiation or amplification of IL-1 and IL-6 release during lethal gram-negative septic shock syndrome.

Clin Orthop, 1989 Nov, (248), 227 - 30
The use of quantitative bacterial counts in open fractures; Moore TJ et al.; Late infections remain a significant complication in the treatment of open fractures, occurring in 2% to 25% of all open fractures . Recently there has been a trend toward early soft-tissue coverage of open fracture wounds . Quantitative bacterial counts have been used primarily by plastic surgeons to assess the viability of primary versus delayed wound closures . Quantitative bacterial counts were obtained in 52 consecutive open fractures in 50 patients to assess the correlation between quantitative bacterial counts and subsequent sepsis, to determine the effect of time to debridement on the quantitative bacterial count, and finally to review the bacteriology of open fractures . Forty-one of the 52 fractures had positive quantitative bacterial counts . Four of eight (50%) fractures with quantitative bacterial counts greater than 10(5) developed late sepsis, while only two of 44 (5%) with quantitative bacterial counts less than 10(5) or negative quantitative counts became infected . There was no correlation between time to debridement and quantitative bacterial count levels . A significant percentage of the positive quantitative bacterial counts were gram-negative organisms.

Am J Reprod Immunol, 1989 Nov-Dec, 21(3-4), 119 - 22
Macrophages in human uteroplacental tissues: a review; Hunt JS; Macrophages are major cellular components of the uterus, the placenta, and the extra-placental membranes throughout pregnancy . Many macrophages in normal uteroplacental tissues are in an activated state as evidenced by their expression of class II major histocompatibility antigens . Activated macrophages are capable of synthesizing and releasing an impressive array of potent "monokines" and other molecules with dramatic effects on receptive cells . Macrophage activation levels can be further enhanced by exposure to either endotoxin from gram-negative organisms or to interferon-gamma from stimulated lymphocytes . Since prostaglandin E2 is one of the products of activated macrophages and can stimulate contraction of smooth muscle, macrophages may be responsible in part for the induction of preterm labor in cases of infection . Thus, understanding the conditions of uncomplicated pregnancy as well as the consequences of local infections requires exploration of the temporal, spatial, and functional characteristics of macrophages residing in uteroplacental tissues.

Plasmid, 1989 Nov, 22(3), 203 - 14
Two mechanisms necessary for the stable inheritance of plasmid RP4; Grinter NJ et al.; Plasmid RP4 is stably maintained in strains of Escherichia coli and other Gram-negative bacteria . Inactivation of the plasmid primase gene (pri) or removal of the PstIC fragment gave RP4 derivatives that are slightly unstably maintained in E . coli . Removal of the Tn 1 multimer resolution system (res and tnpR) did not lead to any detectable plasmid loss . Removal of all three of these regions, however, gave rise to pNJ5000 which is lost at high frequency . We have dissected the mechanisms causing this phenomenon . In contrast to RP4, pNJ5000 accumulates significantly as plasmid multimers in a Rec+ host; in a recA host, multimers are not seen and the plasmid is stably maintained . Multimers therefore appear to form by recA-mediated homologous recombination and cause plasmid instability, perhaps by interfering with partition . We demonstrate a mechanism provided by the PstIC fragment which acts on multimers analogously to the Tn1/3 resolution system on plasmid cointegrates, being effective only when cloned in cis . The loss of pri, on the other hand, can be complemented in trans . Our results are consistent with the view that primase prevents multimers forming (rather than resolving them once formed), perhaps by binding specifically to single-stranded regions of the plasmid and preventing homologous pairing.

S D J Med, 1989 Nov, 42(11), 5 - 8
Eikenella corrodens: an unusual cause of endocarditis in a patient with silent mitral valve prolapse; Hansen LA et al.; A 69-year old man with clinically silent mitral valve prolapse developed infective endocarditis secondary to Eikenella corrodens after dental work . The patient required surgical removal of abscessed teeth and long-term antibiotic therapy . E . corrodens is a gram-negative coccobacillus which normally inhabits the oropharynx, gastrointestinal tract, and upper respiratory tract . The organism can cause cutaneous and abdominal abscesses, meningitis, osteomyelitis, and endocarditis . Patients with mitral valve prolapse and a pre-existent systolic murmur or Doppler echocardiographic evidence of mitral regurgitation should receive prophylactic antibiotics for any procedure associated with a bacteremia . An infection caused by E . corrodens should be considered in patients with fever after dental manipulation or in patients with "culture-negative" endocarditis.

J Antimicrob Chemother, 1989 Nov, 24 Suppl B, 1 - 22
The epidemiology of beta-lactamases; Wiedemann B et al.; Chromosomally mediated species-specific beta-lactamases, as well as plasmid-mediated beta-lactamases, contribute to bacterial resistance to beta-lactam antibiotics . Chromosomally mediated enzymes confer primary resistance to some drugs and secondary resistance, by mutation to over-production of the enzyme . By far the most prevalent and most important of the, more than thirty, described plasmid-mediated beta-lactamases are those of the TEM group . They can be found in nearly all Gram-negative bacterial species of clinical importance . Furthermore these enzymes have changed their specificity by mutation so that recently described TEM enzymes hydrolyse even third generation cephalosporins . Although there is no change in the quantity of these enzymes, as far as the percentage of producer strains in any species is concerned, there has been a change in quality . The enzymes are further distributed to new species and hydrolyse more so-called 'stable' beta-lactam compounds.

Rev Infect Dis, 1989 Nov-Dec, 11 Suppl 7, S1669 - 90
Drug delivery to the central nervous system: general principles and relevance to therapy for infections of the central nervous system; Scheld WM; Drug delivery to the central nervous system (CNS) is of vital concern to the therapy for primary CNS disorders and the development of drug neurotoxicity . The factors influencing drug entry into the CSF include the status of the blood-brain barrier (BBB) and lipid solubility, molecular weight, pKa, protein binding, and removal of the drug from the CSF by an exit pump in the choroid plexus . The most important of these factors is the status of the complex BBB systems . The morphologic equivalent of the BBB and its specialized functions (e.g., transport of D-glucose, amino acids, and ions) are discussed in depth . Methods developed for increasing drug delivery to the CNS by circumvention and/or manipulation of the BBB have included direct injection into the CSF, administration of prodrugs or chemical delivery systems, or reversible "opening" of the BBB by hyperosmotic agents, pentylenetetrazole, etoposide, DMSO, or other agents . The relevance of these general principles to selected examples of CNS infections (i.e., gram-negative aerobic bacillary meningitis and subacute encephalopathy associated with AIDS) is emphasized.

Haematologica, 1989 Nov-Dec, 74(6), 583 - 5
Pefloxacin in the treatment of gram-negative infections in patients with hematologic diseases; Micozzi A et al.; Quinolones are active against gram-negative strains . They are commonly used for selective intestinal decontamination in patients with hematologic malignancies and prolonged neutropenia due to chemotherapy . In our open study we used pefloxacin, a new fluoroquinolone, for the treatment of fifteen documented gram-negative infections in hematologic patients . Thirteen patients were mildly neutropenic, and in nine cases they received oral treatment as non-hospitalized patients . Cure was achieved in fourteen cases, with microbiological eradication of the offending pathogen.

Infection, 1989 Nov-Dec, 17(6), 382 - 4
T lymphocyte disorder after Capnocytophaga ochracea endocarditis; Gormand F et al.; Capnocytophaga species are gram-negative rods which may cause disease in both non-immunocompromised and immunocompromised hosts . We describe a case of endocarditis due to Capnocytophaga ochracea in a non-immunocompromised patient with a decrease of blood CD4/CD8 ratio and lymphocyte proliferative response to ConA during infection . In vitro experiments showed that C . ochracea decreased lymphocyte proliferation to mitogens (ConA, PHA), cell surface CD4 antigen and IL2 receptor expression on peripheral blood mononuclear cells (PBMC) from normal volunteers.

J Biol Chem, 1989 Oct 15, 264(29), 17462 - 8
Protein secretion by gram-negative bacteria . Characterization of two membrane proteins required for pullulanase secretion by Escherichia coli K-12; d'Enfert C et al.; Pullulanase secretion in Escherichia coli depends on the expression of a MalT-regulated operon called pulC . Characterization of the first two genes of this operon showed that they encode, respectively, a 31,000-Da protein (PulC) and a 70,600-Da protein (PulD) which has a putative signal peptide and that these two proteins are required for pullulanase secretion . The analysis of alkaline phosphatase hybrid proteins generated by TnphoA mutagenesis of pulC and pulD showed that both PulC and PulD contain export signals which can direct the alkaline phosphatase segment of the hybrids across the inner membrane . A representative PulC-PhoA hybrid protein fractionated mainly with the inner membrane upon isopycnic sucrose gradient centrifugation of membrane vesicles . This, together with sequencing data, suggests that PulC is an inner membrane protein . Antibodies raised against a purified PulD-PhoA hybrid protein were used to show that PulD was enriched in low density outer membrane vesicles.

CLAO J, 1989 Oct-Dec, 15(4), 268 - 70
Contact lens related corneal ulcers at the Ohio State University Hospitals 1983-1987; Derick RJ et al.; We reviewed the records of patients with ulcerative keratitis associated with contact lens wear admitted to The Ohio State University Hospitals from January 1, 1983 through December 31, 1987 . Of 127 cases of infectious ulcerative keratitis, 25 (19%) were associated with the use of contact lenses . Of these 25, 21 cases (84%) were associated with the use of soft lenses (40% aphakic, 40% cosmetic) . Seventy-six percent (19 of 25) were culture-positive; Pseudomonas was the most common isolate (12 of 19, 63% of culture-positive cases) . When compared with daily wear soft contact lenses, cosmetic and aphakic extended wear contact lens related ulcers were associated with a delay in definitive diagnosis and a worse prognosis . Patching and steroid use were associated with more severe ulcers . Gram stain findings were of value only when gram-negative rods were noted . The findings emphasize the need for patients to understand: 1) the risks of extended wear; 2) the necessity of removing the lenses at the first sign of irritation; and 3) the importance of having prompt, intensive care readily available.

Gastroenterology, 1989 Oct, 97(4), 1025 - 7
Pancreatic duct-portal vein fistula; Willis SM et al.; We describe a case of pancreatic duct-portal vein fistula discovered by endoscopic retrograde cholangiopancreatography, with confirmatory computed tomography and angiographic studies . The fistula was associated with chronic pancreatitis that was complicated by recurrent gram-negative bacteremia and peripheral subcutaneous fat necrosis, although pancreatic pseudocyst was not present . Even though surgical treatment was recommended, medical complications precluded surgery and the patient recovered with supportive therapy only . Follow-up endoscopic retrograde cholangiopancreatography showed spontaneous closure of the fistulous connection.

Circ Shock, 1989 Oct, 29(2), 77 - 92
Effect of antioxidants in experimental Escherichia coli septicemia; Broner CW et al.; Free radicals have been implicated in the pathogenesis of gram-negative bacterial sepsis . We assessed the effectiveness of antioxidants and chelators to alter oxidative injury in established severe experimental Escherichia coli septicemia . One hour after challenge by intraperitoneal injection of bacteria, 36 rabbits were treated with moxalactam and randomized in sets of three to receive either placebo, superoxide dismutase (SOD), or a combination of antioxidants and chelators consisting of SOD, sodium thiosulfate, alpha-tocopherol, deferoxamine, and diethyldithiocarbamate . Throughout the course of treatment, levels of bacteremia and endotoxemia were similar among the three experimental groups . Neither antioxidant-treated group was significantly different from the control group in mean arterial blood pressure, leukocyte count, platelet count, core temperature, blood lactate, oxygenation or survival . Arterial pH and {HCO3-} were significantly lower in the antioxidant combination group compared to the control and SOD groups (P less than .01) . In this model, antioxidant and chelator therapy does not substantially ameliorate established septicemia.

Circ Shock, 1989 Oct, 29(2), 133 - 42
Gram-negative endotoxemia: effects on cardiac Na-Ca exchange and stoichiometry; Hale CC et al.; We studied the acute effects of gram-negative endotoxemia on Na-Ca exchange activity and stoichiometry in cardiac sarcolemmal (SL) vesicles isolated from pentobarbital-anesthetized dogs . Dogs were given either endotoxin (ET; 1.5 mg/kg IV) or saline vehicle (C; n = 4 dogs/group) . Characteristic of endotoxemia, endotoxin produced a decrease in mean arterial pressure from 120 to 60 mmHg, an increase in packed cell volume from 38% to 60%, and an increase in heart rate from 130 to 190 bpm . After 2 hr, hearts were removed and SL vesicles were prepared from left and right ventricular tissue . For ET and C, Na-dependent Ca2+ uptake (left ventricle) was 3.13 and 3.44 (2 sec) and 18.60 and 19.42 (60 sec) nmole Ca2+/mg protein, respectively; ET group values were not significantly different from corresponding C values in either left or right ventricles . The stoichiometry of Na-Ca exchange was determined in left ventricular vesicles by a previously described thermodynamic approach utilizing a K+-valinomycin gradient opposed by Na+ equilibrium potentials (Reeves and Hale: J Biol Chem 259:7733-7739, 1984) . The stoichiometry of exchange of Na+ for Ca2+ was 2.84 +/- 0.09 and 2.74 +/- 0.14 for ET and C, respectively . We conclude that during the developmental phase (2 hr) of endotoxemia, there were no ET-mediated changes in cardiac Na-Ca exchange activity in SL vesicles from either left or right ventricular tissue and the exchange process remained electrogenic with a stoichiometry of 3Na+ for 1Ca2+.

J Am Podiatr Med Assoc, 1989 Oct, 79(10), 497 - 9
Use of ciprofloxacin in podiatric medicine; LeFrock JL; Ciprofloxacin is the first of the new class of antibiotics known as fluoroquinolones to be approved for use in skin, skin structure, and bone and joint infections . It has an extremely broad spectrum and is particularly effective against traditionally resistant gram-negative rods . As an oral agent, it is as effective as parenteral drugs against a variety of organisms and diseases . Its spectrum, pharmacokinetics, and podiatric indications are reviewed.

Eur J Haematol, 1989 Oct, 43(4), 309 - 13
Idarubicin in combination with intermediate-dose cytarabine in the treatment of refractory or relapsed acute leukemias; Carella AM et al.; 13 patients with refractory or relapsed acute lymphoblastic leukemia (ALL) and 7 patients with acute myeloid leukemia (AML) were treated with a regimen that included idarubicin 12 mg/m2 intravenously daily for 3 d plus cytarabine 2 g/m2 by infusion over 3 hours daily for 3 d . There were 10 remissions (ALL:7; AML:3) in the 15 relapsed patients and 4 (ALL:3) in the 5 patients with primary refractory disease . Severe myelosuppression was observed in all patients . Toxicity of this regimen caused nausea and vomiting, stomatitis, infections and/or liver enzymes increase . Cardiac toxicity was not observed . 2 patients died in aplasia of Gram-negative septicemia and brain hemorrhage . In conclusion, the combination of idarubicin and intermediate-dose cytarabine (IDARA-C) seems to be highly effective and sufficiently well-tolerated for the treatment of refractory and relapsed acute leukemias.

Circ Shock, 1989 Oct, 29(2), 93 - 106
Changes in anti-endotoxin-IgG antibody and endotoxaemia in three cases of gram-negative septic shock; Barclay GR et al.; Circulating endotoxin levels and IgG antibodies to a range of Gram-negative bacterial lipopolysaccharides (LPS) (endotoxins) of different sizes and structures were measured daily in three cases of septic shock . There was an inverse relationship between endotoxin levels and cross-reactive antibodies to the core glycolipid (CGL) region of lipopolysaccharide . This suggests that antibody to LPS-CGL was initially consumed by a superabundance of endotoxin, and that a resurgence of intrinsic anti-LPS-CGL antibody levels may be associated with a reduction of circulating endotoxin . The implications of these findings for passive antibody therapy of septic shock are discussed.

J Pharm Pharmacol, 1989 Oct, 41(10), 694 - 9
Use of strips containing tetracycline hydrochloride or metronidazole for the treatment of advanced periodontal disease; Deasy PB et al.; Strips containing tetracycline hydrochloride or metronidazole 25% in polyhydroxybutyric acid as a biodegradable polymer matrix, showed sustained release in simulated gingival fluid pH 6.6 at 37 degrees C . When evaluated in patients suffering from advanced periodontal disease, the greatest response to therapy was observed with tetracycline hydrochloride strips inserted into periodontal pockets at four-day intervals for 16 days, compared with an untreated control group . A reduction in plaque index, gingival index and pocket depth was observed . A favourable alteration occurred in the microbial flora of treated pockets with an increase in the proportion of cocci and decrease in gram-negative rods, fusiforms and spirochetes . Metronidazole strips or root-planning tended not to be as effective . The clinical improvement produced by each treatment was not maintained when treatment was terminated.

J Hosp Infect, 1989 Oct, 14(3), 217 - 25
Nosocomial gram-negative bloodstream isolates: a comparison of in vitro antibiotic potency; Stanley GL et al.; Nosocomial bloodstream infections add to the morbidity, mortality and length of hospitalization that is attributed to the underlying diseases alone . We have compared the in vitro potency of fifteen antibiotics against 136 isolates from clinically significant nosocomial gram-negative bacteraemias . Ciprofloxacin was the most potent antibiotic and had the broadest spectrum of activity (98% of isolates susceptible, MIC90 range: 0.06-0.5 micrograms ml-1) . We subjected all isolates to beta-lactamase induction but antibiotic susceptibility was unaffected by this procedure.

Vet Hum Toxicol, 1989 Oct, 31(5), 422 - 6
Effects of polymyxin B on selected features of equine carbohydrate overload; Raisbeck MF et al.; Gram negative endotoxins play a contributory role in the syndrome which results from over consumption of carbohydrates by horses and ponies . Since the antibiotic polymyxin B exerts a direct anti-endotoxin effect by chemically modifying the active lipid A moiety of endotoxin, it might be expected to protect horses after carbohydrate overload and provide a new therapeutic and experimental tool for this condition . The present study was undertaken to evaluate the effect of polymyxin B on hemostatic, hemodynamic, acid-base, and clinical aspects of the syndrome resulting from carbohydrate overload . Experimentally-induced carbohydrate overload resulted in lactic acidosis, hypercoagulability, hypovolemic shock and lameness . Although there was a slight delay in the onset of clinical signs resulting from experimental carbohydrate overload in treated animals, polymyxin B administered iv at 2.5 mg/kg every 6 hr failed to significantly ameliorate the coagulopathy, acidosis, lameness and shock induced by alimentary carbohydrate overload.

J Antimicrob Chemother, 1989 Oct, 24(4), 523 - 31
Use of the Malthus Microbial Growth Analyser to study the post antibiotic effect of antibiotics; Gould IM et al.; The post antibiotic effect (PAE) of ceftazidime, ciprofloxacin, mecillinam and imipenem alone and in combination against Gram-negative bacteria was assessed by a new technique using a Malthus Microbial Growth Analyser . Ciprofloxacin gave the most prolonged and consistent PAE (1.3-2.9 h) and imipenem also gave a significant PAE against some bacterial strains (up to 1.3 h) . The PAE of both antibiotics was dependent on concentration . The PAE of combinations of ciprofloxacin and imipenem often showed less PAE than was present with either agent alone . Ceftazidime gave no significant PAE (-1.5-0.4 h), though mecillinam consistently gave a short PAE (approximately 0.5 h) against Escherichia coli . The new method allows for the rapid and labour saving evaluation of PAE . We believe that further studies on PAE of antibiotic combinations are desirable.

Eur J Clin Microbiol Infect Dis, 1989 Oct, 8(10), 901 - 3
Decontaminating efficacy of ciprofloxacin in an animal model; van Ogtrop ML et al.; A study was performed to assess the suppressive effect of ciprofloxacin on the aerobic gram-negative flora of the gastrointestinal tract in mice in relation to its effect on colonization resistance . Ciprofloxacin exhibited a potent decontaminating effect, with the maximum effect being reached at a dose of 8 mg/kg/day . At this dose aerobic gram-negative rods were eliminated from the gastrointestinal tract within 24h without any adverse influence on the parameters of colonization resistance measured.

J Gen Microbiol, 1989 Oct, 135 ( Pt 10), 2601 - 9
Oxygen inhibition of globin gene transcription and bacterial haemoglobin synthesis in Vitreoscilla; Dikshit KL et al.; A soluble dimeric haemoprotein, structurally and functionally similar to plant and animal haemoglobins, is found in the Gram-negative aerobic bacterium Vitreoscilla sp., strain C1 . Vitreoscilla haemoglobin (VtHb) increases in concentration when the cells are exposed to hypoxic conditions . The globin part of VtHb is encoded by a single gene (vgb) . An RNA transcript, approximately 500 bases long, specific for vgb was detected after Northern hybridization . The relative amount of this mRNA increased in cells grown at low levels of oxygen . Two enzymes important for haemoglobin function are delta-aminolaevulinic acid synthase (ALAS), which is necessary for haem biosynthesis, and NADH-methaemoglobin reductase, which is necessary to keep VtHb in the physiologically functional ferrous state . An increase in ALAS specific activity under hypoxic conditions preceded the increased haem production . Cellular reductase content also increased when the VtHb increased in cells grown under hypoxic conditions . The ratio of cellular reductase activity to VtHb content remained relatively constant in cells grown under a variety of conditions . The data suggest that in Vitreoscilla the transcription of the globin gene and the biosynthesis of two enzymes important for VtHb function are regulated by oxygen.

Presse Med, 1989 Sep 23, 18(29), 1419 - 22
{Preventive treatment of infection caused by gram-negative bacteria using anti-lipopolysaccharide antibodies . Evaluation in patients with aplasia}; Lecomte F et al.; The prophylaxis of severe Gram-negative infections with human antiserum to lipopolysaccharide (LPS) was evaluated in a randomised study of 60 patients with therapeutic aplasia for leukaemia . The antiserum was found to be ineffective in preventing Gram-negative infections . The levels of anti-LPS antibodies showed that passive immunization was obtained in only one half of the patients . These disappointing results warrant further investigations to evaluate the effectiveness of this prophylactic treatment.

Am J Med, 1989 Sep 11, 87(3B), 27S - 33S
Efficacy of antithrombin III supplementation in animal models of fulminant Escherichia coli endotoxemia or bacteremia; Emerson TE Jr et al.; Plasma antithrombin III (ATIII) levels decrease early during gram-negative septicemia, and even a moderate decrease in this major inhibitor of the coagulation system is associated with serious disseminated intravascular coagulation (DIC) . Herein the efficacy of high-dose (at least 250 units/kg) ATIII supplementation in animal models of Escherichia coli endotoxemia or bacteremia is reported . An endotoxemic rat model demonstrated that: (1) DIC occurs very early, before the appearance of deleterious cardiovascular abnormalities; (2) ATIII prophylaxis attenuates DIC, metabolic dysfunction, and organ damage; (3) ATIII prophylaxis increases permanent survival; (4) ATIII treatment one hour after endotoxin challenge attenuates DIC, metabolic dysfunction, and organ damage, although not as well as when given prophylactically, and survival is not increased . An endotoxemic sheep pulmonary dysfunction model demonstrated that: (1) ATIII prophylaxis prevents the typical decrease in arterial oxygen partial pressure; (2) ATIII prophylaxis combined with alpha-1-proteinase inhibitor significantly attenuates indices of pulmonary dysfunction . An E . coli bacteremic baboon model demonstrated that ATIII prophylaxis and treatment significantly attenuate indices of DIC and organ damage and prevent death in an otherwise completely lethal dose bacterial challenge . In conclusion, prophylactic treatment with high doses of ATIII may be efficacious in disease states of impending disseminated intravascular coagulation, such as primary or secondary gram-negative septicemia.

Hosp Formul, 1989 Oct, 24(10), 598, 607 - 9, 612
Gentamicin blood levels: ordered too soon and too often; Averbuch M et al.; Records of 100 consecutive patients in whom gentamicin blood levels (GBLs) had been measured were reviewed to assess the relation between GBL, disease course, and outcome . Thirty-two patients discontinued treatment, 16 within 72 hours because of negative cultures or improved clinical course, and 16 within 96 hours because susceptibility tests indicated that another antibiotic would be more appropriate . Of the remaining 68 patients who continued treatment, 21 recovered from their infection despite low peak or low peak and trough gentamicin levels, at least partially attributable to treatment with multiple antibiotics . Renal function deteriorated in only 1 of 10 patients with normal renal function and a high GBL . Our findings of early discontinuation of gentamicin, low concentrations with good outcome, and rare nephrotoxicity indicate wasteful overuse of GBLs . To decrease patient care costs without adversely affecting outcome, gentamicin measurements should be delayed for 72 to 96 hours--until culture results become available--except in patients with decreased renal function, documented gram-negative sepsis, leukopenia, or severe burns.

Ther Drug Monit, 1989 Sep, 11(5), 574 - 9
Serial pharmacokinetic dosing of aminoglycosides: a community hospital experience; Hoffa DE; The aminoglycoside antibiotics are useful in the treatment of gram-negative bacillary infections but are potentially toxic . A method to maximize their therapeutic benefit while minimizing their risk of toxicity is desirable . Serial pharmacokinetic dosing has been proposed as a method to achieve these goals . An audit was conducted comparing optimal dosing of aminoglycosides by physicians of a community hospital using a serial pharmacokinetic dosing service versus its nonuse . Optimal dosing was 81% (81/101) of trough-peak pairs using the service versus 17% (28/161) not using the service (p less than 0.001) . This difference was due to greater achievement of therapeutic peak levels with the service (96%, 97/101) than without (32%, 52/161) . There was no difference in toxic trough level occurrence, with 15% (15/101) occurrence with the service and 17% (27/161) occurrence without its use (p greater than 0.05) . Clinical nephrotoxicity occurred 0% (0/49) of the time with the use of the service versus 7% (6/88) of the time without its use (p greater than 0.05) . The data presented here demonstrate that serial pharmacokinetic dosing of aminoglycosides results in the achievement of therapeutic peak levels in most patients . An understanding of aminoglycoside nephrotoxicity and pharmacokinetics allows the subsequent adjustment of dosing, if necessary, to avoid nephrotoxicity in most patients.

J Vet Pharmacol Ther, 1989 Sep, 12(3), 296 - 301
The effect of avilamycin in the control of stress-induced post-weaning diarrhoea in piglets; Kyriakis SC; Avilamycin, an oligosaccharide antibiotic with growth-promoting properties in pigs, has proved to be effective in controlling stress-induced post-weaning diarrhoea in piglets, caused mainly by Escherichia coli . The present study includes two trials, in which 400 newly-weaned piglets were used (200/trial) . The following five different treatments were tested; 0, 40 and 80 p.p.m . avilamycin, 50 p.p.m . olaquindox and 100 p.p.m . apramycin . In each trial there were four pens (each with five females and five males) per treatment . Avilamycin when given at 80 p.p.m . reduced average daily diarrhoea score (ADDS) and mortality, and improved liveweight gain and feed conversion ratio (FCR), compared with the untreated controls, the 40 p.p.m . avilamycin and the 50 p.p.m . olaquindox (P less than 0.05) treatments . The overall performance of 40 p.p.m . avilamycin and 50 p.p.m . olaquindox was similar . The results indicate that avilamycin at the dose level of 80 p.p.m . in the starter feed can control post-weaning diarrhoea of piglets and prevent loss of productivity . Nevertheless, the antibiotic apramycin, whose spectrum of activity is mainly against the Gram-negative bacteria, given at the therapeutic level of 100 p.p.m., was more effective than any other experimental treatment (P less than 0.05), except for ADDS and FCR which were not significantly different from that of avilamycin 80 p.p.m.

J Cardiovasc Surg (Torino), 1989 Sep-Oct, 30(5), 870 - 2
Infection of an abdominal aortic aneurysm from an appendix abscess; Jewkes AJ et al.; We report a case of gram negative anaerobic infection developing in an abdominal aortic aneurysm due to direct involvement by an appendix abscess . Diagnosis, management and successful treatment of this unusual and often lethal condition are discussed.

Biomed Environ Mass Spectrom, 1989 Sep, 18(9), 741 - 52
Characterization of new bacterial transformation products of 1,1,1-trichloro-2,2-bis-(4-chlorophenyl) ethane (DDT) by gas chromatography/mass spectrometry; Masse R et al.; The microbial transformation of DDT, DDD and DDE was studied in Gram-negative strain B-206 and a number of phenolic metabolites were identified as the trimethylsilyl derivatives in the bacterial extracts by gas chromatography/mass spectrometry . The major metabolites of DDT were DDD, DDE, DDMU, 1,1,1-trichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-chlorophenyl) ethane, 1,1,1-trichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-hydroxyphenyl) ethane, and 1,1,1-trichloro-2,2-bis-(2-hydroxy-4-chlorophenyl) ethane . Conversely, DDD was mainly degraded into DDE, 1,1-dichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-chlorophenyl) ethane and 1,1-dichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-hydroxyphenyl) ethane . Finally, DDE was transformed into DDMU, 1,1-dichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-chlorophenyl) ethylene, 1,1-dichloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'hydroxyphenyl) ethylene and 1-chloro-2-(2-hydroxy-4-chlorophenyl)-2-(4'-chlorophenyl) ethylene . The phenolic metabolites exhibited {M - TMSCl}+., {M - HCl - TMSCl}+ . and/or {M - HCl - TMSCl - Me}+ fragment ions which reflect the presence of an ortho hydroxyl group in these molecules . Other mass spectral features used to determine their structure are presented and a metabolic scheme accounting for their formation is proposed.

Biometrics, 1989 Sep, 45(3), 797 - 816
A discrete-time model of the acquisition of antibiotic-resistant infections in hospitalized patients; Garber AM; Antibiotic use is thought to promote bacterial antibiotic resistance by selectively inhibiting the growth of sensitive strains . This study investigates the relation between antibiotic use and the propagation of antibiotic-resistant hospital-acquired infections due to gram-negative bacteria in a population of hospitalized patients . It treats infection spread and hospital mortality as a Markov process, in which the transition probabilities are logistic functions of a set of personal and hospital characteristics . Data from a university hospital are used to derive the parameters of the model.

Am J Pathol, 1989 Sep, 135(3), 489 - 97
Endotoxin-induced shock in the rat . A role for C5a; Smedegard G et al.; Administration of endotoxin from gram-negative bacteria to rats results in systemic hypotension, an increased hematocrit, and decreased numbers of circulating leukocytes (polymorphonuclear), monocytes, and platelets . These potentially lethal physiologic changes may be partially attributed to complement activation and generation of anaphylatoxins by the endotoxin (LPS) . We demonstrated an elevation in the plasma levels of both C3a and C5a in LPS-treated rats . Injection of 5 micrograms C5ades Arg (rat) into rats produced effects similar to those induced by LPS, including decreased mean arterial pressure (systemic hypotension) and decreased numbers of circulating polymorphonuclear leukocytes, monocytes, and platelets . Unlike the response to LPS, C5a did not increase the hematocrit, indicating little effect on vascular permeability at the doses used . When LPS-treated animals were pretreated with F(ab')2 fragments of rabbit anti-rat C5a, no changes were measured in the circulating cell counts compared with LPS alone; however a significant improvement in the mean arterial pressure and a decrease in hematocrit was observed . We conclude that LPS-induced (septic) shock in the rat may result, in part, from the effects of complement activation and particularly from the generation of C5a . The influence of C5a on the LPS effect in the rat appears to enhance both the hypotensive (mean arterial pressure) and vascular permeability (hematocrit) responses . These results appear to support and confirm earlier observations that anti-human C5a increased survival in a septic-shock monkey model by eliminating circulating C5a and presumably thereby reducing the effects of endotoxin on blood pressure . Our results demonstrate that C5a plays a significant role in the hemodynamic changes associated with endotoxin-induced shock . Neutralization of C5a with specific antibodies may reduce the hypotensive response to endotoxin sufficiently to prevent lethal septic shock both in animals and in man.

Obstet Gynecol, 1989 Sep, 74(3 Pt 2), 465 - 8
Fetal death from sepsis following a reassuring intrapartum fetal acoustic stimulation test; Strong TH Jr et al.; An intrapartum fetal death within 20 minutes of a reassuring acoustically stimulated fetal heart rate acceleration is reported . The cause of death in this instance was congenital pneumonia, gram-negative sepsis, and meconium aspiration . Umbilical cord pH values obtained at delivery did not demonstrate asphyxia (ie, low pO2, high pCO2, and low pH), but suggested a metabolic acidosis typical of sepsis.

Microb Pathog, 1989 Sep, 7(3), 175 - 88
Characterization of lipid-modified immunogenic proteins of Treponema pallidum expressed in Escherichia coli; Schouls LM et al.; This study describes the sequence of the immunodominant Treponema pallidum surface protein TpD and its expression in Escherichia coli . The translated TpD DNA sequence revealed the presence of a putative site for lipid-modification downstream from the signal sequence of this membrane protein . Growth of TpD-expressing E . coli in the presence of radioactive palmitic acid revealed that TpD was lipid-modified . Three other, previously characterized cloned proteins of T . pallidum were also lipid-modified . The N-termini of two of three sequenced T . pallidum proteins contain a tetrapeptide sequence characteristic for lipoproteins in Gram-negative bacteria: Leu-X-Y-Cys . Only TpD differed from this consensus sequence in the substitution of the first residue by Phe . The apparent high incidence of lipoproteins among E . coli recombinants expressing T . pallidum antigens suggest an important role of lipoproteins in the induction of humoral immunity during syphilitic infection.

Infection, 1989 Sep-Oct, 17(5), 333 - 7
Resistance to third generation cephalosporins: the current situation; Pechere JC; Newer beta-lactam antibiotics, notably the third generation cephalosporins (3 GC) have been designed for providing high intrinsic potency against a large variety of microorganisms . Bacterial resistance can occur however, and nowadays, clinicians are concerned by novel situations where even most recently developed compounds can be ineffective . A first situation is generated by bacteria which produce great amounts of chromosomal cephalosporinase . The resistance emerges during therapy, in hospital isolates which are classified as susceptible with conventional susceptibility testing . The prevalence of 3 GC resistance among these gram-negative rods with inducible beta-lactamase seems to increase in some institutions but the significance of susceptibility testing in this regard is doubtful . It is probably more important to note that the prevalence of gram-negative rods with inducible beta-lactamases remains stable . A second problem arose with the abrupt development of plasmid mediated beta-lactamases markedly active against 3 GC . This resistance is underestimated because some strains fall into susceptibility range of 3 GC as determined by MICs or inhibition zone sizes . These extended spectrum enzymes are now distributed over four continents and represent a growing threat.

Acta Chir Scand, 1989 Sep, 155(9), 451 - 4
Influence of splenectomy, partial splenectomy and splenic artery ligation on E . coli sepsis in rats; Andersson R et al.; Male Sprague-Dawley rats were allocated to four groups--sham operation, partial splenectomy, splenic artery ligation or total splenectomy, and 4 weeks after the operation 3 x 10(8) colony-forming units of Escherichia coli were injected intraperitoneally . Among the splenectomized rats the mortality was significantly (p less than 0.02) increased compared with the controls, while both partial splenectomy and splenic artery ligation did not influence survival . Blood clearance and organ (liver, spleen and lungs) uptake of intravenously injected, radiolabelled, heat-killed E . coli were determined 1 hour after the intraperitoneal challenge . Splenectomy caused a significant decrease in blood clearance . Splenic uptake of radiolabelled E . coli was significantly reduced following partial splenectomy and splenic artery ligation . The splenic operations increased hepatic uptake expressed per gram tissue . Splenectomy thus resulted in reduced blood clearance and increased mortality in Gram-negative sepsis, while the reduced splenic uptake following partial splenectomy or splenic artery ligation did not influence blood clearance of E . coli or mortality.

Bioorg Khim, 1989 Sep, 15(9), 1232 - 8
{Chemical-enzymatic synthesis and cloning of DNA, coding the signal for secretion of proteins in gram-negative bacteria}; Dobrynin VN et al.; Chemical-enzymatic synthesis of an artificial gene encoding leader peptide and 22 N-terminal amino acids of mature carboxypeptidase G2 from Pseudomonas sp . followed by enterokinase signal sequence (Asp4Lys) has been accomplished . The resulted DNA was fused with semi-synthetic gene coding for polypeptide 4-157 of mature human tumour necrosis factor (TNF) and then placed under control of early promoters of T7 bacteriophage . The expression products of the construct obtained was analysed using anti-TNF anti-serum . In E.coli leader peptide was cleaved off during translocation through inner membrane and the resultant product was found in membrane fraction.

Pediatr Infect Dis J, 1989 Sep, 8(9 Suppl), S124 - 7; discussion S128-32
Aztreonam in the management of gram-negative infections in penicillin-allergic patients: a review; Saxon A; Penicillins and cephalosporins, which are well-tolerated by most patients, can result in hypersensitivity reactions . This factor not only limits the use of beta-lactam antibiotics but also presents challenges to the selection of alternative drugs because of the potential for cross-reactivity . Evidence suggests that aztreonam, the prototype of the new monobactam class of beta-lactam antibiotics, may be a safe alternative antibiotic for managing Gram-negative infections in penicillin-allergic patients.

Pediatr Infect Dis J, 1989 Sep, 8(9 Suppl), S120 - 3; discussion S128-32
Safety and toxicity profile of aztreonam; Chartrand SA; Aztreonam is the first monocyclic beta-lactam antibiotic released for clinical use . Extensive toxicity and safety data for aztreonam in animals, healthy volunteers and adult patients have been accumulated previously; recently these studies have been extended to children . Overall the incidence of adverse clinical reactions caused by aztreonam is similar to or lower than that caused by comparison drugs . There is no evidence that aztreonam causes significant ototoxicity or nephrotoxicity; biochemical and hematologic abnormalities are rarely significant . Compared with the broad spectrum cephalosporins, aztreonam causes less disruption of normal gastrointestinal flora and consequently a lower incidence of diarrhea . Aztreonam does not displace bilirubin from albumin and penetrates readily into cerebrospinal fluid . Because of negligible immunologic cross-reactivity with other beta-lactams, aztreonam has been safely administered to patients with IgE-mediated penicillin hypersensitivity . These data suggest that aztreonam should be safe and well-tolerated in infants and children with infections caused by susceptible Gram-negative bacteria.

J Trauma, 1989 Sep, 29(9), 1252 - 7
Adrenergic mechanisms contribute to alterations in regional perfusion during normotensive E . coli bacteremia; Fantini GA et al.; The complex role of adrenergic mechanisms in the pathogenesis of sepsis and shock remains incompletely understood . Recent reports have suggested that adrenergic mechanisms may play an important modulatory role in arachidonate metabolism during early sepsis, specifically in the synthesis of the vasoactive metabolites thromboxane A2 and prostacyclin . The purpose of the present set of experiments was to evaluate the extent to which adrenergic mechanisms contribute to early alterations in regional perfusion during Gram-negative bacteremia, using the mixed alpha adrenergic receptor antagonist phenoxybenzamine hydrochloride . Male Sprague-Dawley rats received a 3-hour continuous intraarterial infusion of either saline (n = 7) or 6.6 +/- 0.4 x 10(8) live E . coli colony forming units (n = 7), after which regional perfusion was determined using 51Cr-labelled microspheres (16.5 +/- 0.1 micron) . Significant reductions (p less than 0.05) in blood flows to the renal, gastric, cecal, pancreatic, and splenic beds, as well as a reduction (p less than 0.05) in the calculated portal venous flow, were observed in the bacteremic group while mean arterial blood pressure remained unchanged . Further experiments conducted in 14 rats with preexisting alpha adrenergic receptor blockade revealed relative preservation of renal, gastric, cecal, and portal venous blood flows during bacteremia . These data identify alterations in regional perfusion during early E . coli bacteremia occurring before changes in systemic arterial blood pressure, and indicate that adrenergic mechanisms participate in the modulation of such.

Infect Dis Clin North Am, 1989 Sep, 3(3), 613 - 24
Imipenem and aztreonam; Sobel JD; To date, clinical use of aztreonam has been remarkably restrained and widespread enthusiasm for addition to hospital formularies similarly moderate . Its most accepted indication appears to be in gram-negative bacillary infections in the penicillin-allergic individual where one might ordinarily use an extended spectrum penicillin or a third generation cephalosporin . In all individuals its clinical efficacy appears similar to that of the aminoglycosides, not clinically superior but offering a higher therapeutic index by virtue of its lack of nephrotoxicity and ototoxicity . Its second application, therefore, is as an aminoglycoside-sparing agent even for multiresistant gram-negative bacillary infections, and although by no means inexpensive it offers an efficacious and safe alternative to aminoglycosides in the elderly and other patients with renal insufficiency . Aztreonam can be used effectively in combination with non-beta-lactam agents for mixed infections, even though it is not synergistic in vitro with vancomycin and clindamycin.

J Bacteriol, 1989 Sep, 171(9), 5005 - 11
Demonstration of rare protein in the outer membrane of Treponema pallidum subsp . pallidum by freeze-fracture analysis; Walker EM et al.; The surface of Treponema pallidum subsp . pallidum (T . pallidum), the etiologic agent of syphilis, appears antigenically inert and lacks detectable protein, as judged by immunocytochemical and biochemical techniques commonly used to identify the outer membrane (OM) constituents of gram-negative bacteria . We examined T . pallidum by freeze-fracture electron microscopy to visualize the architecture of its OM . Treponema phagedenis biotype Reiter (T . phagedenis Reiter), a nonpathogenic host-associated treponeme, and Spirochaeta aurantia, a free-living spirochete, were studied similarly . Few intramembranous particles interrupted the smooth convex and concave fracture faces of the OM of T . pallidum, demonstrating that the OM of this organism is an unusual, nearly naked lipid bilayer . In contrast, the concave fracture face of the OM of S . aurantia was densely covered with particles, indicating the presence of abundant integral membrane proteins, a feature shared by typical gram-negative organisms . The concentration of particles in the OM concave fracture face of T . phagedenis Reiter was intermediate between those of T . pallidum and S . aurantia . Similar to typical gram-negative bacteria, the OM convex fracture faces of the three spirochetes contained relatively few particles . The unique molecular architecture of the OM of T . pallidum can explain the puzzling in vitro properties of the surface of the organism and may reflect a specific adaptation by which treponemes evade the host immune response.

Antimicrob Agents Chemother, 1989 Sep, 33(9), 1428 - 34
Purification, toxicity, and antiendotoxin activity of polymyxin B nonapeptide; Danner RL et al.; Polymyxin B, a relatively toxic antibiotic, has potent endotoxin-neutralizing properties that may be beneficial as adjunctive therapy in gram-negative sepsis . Polymyxin B nonapeptide (deacylated polymyxin B) is devoid of antibiotic activity but retains the capacity to disorganize the outer membrane of gram-negative bacteria . To evaluate the potential therapeutic usefulness of this derivative, we produced purified polymyxin B nonapeptide, tested its in vivo toxicity in animals, and evaluated its in vitro antiendotoxin activity . Effectiveness as an antiendotoxin agent was assessed by examining the ability of polymyxin B nonapeptide to block the enhanced release of toxic oxygen radicals induced by lipopolysaccharide in human neutrophils (priming) . In vivo, at doses of 1.5 and 3.0 mg/kg, polymyxin B nonapeptide did not exhibit the neuromuscular blocking, neurotoxic, or nephrotoxic effects that were observed with polymyxin B sulfate . Both polymyxin B and polymyxin B nonapeptide inhibited lipopolysaccharide-induced neutrophil priming in a concentration-dependent manner, but the parent compound, polymyxin B, was 63 times more effective on a weight basis . The inhibitory activity of both compounds, however, diminished rapidly when they were added after the start of the lipopolysaccharide-neutrophil incubation . We conclude that polymyxin B nonapeptide is less toxic than polymyxin B and, at the doses tested, lacks the neurotoxicity and nephrotoxicity of the parent compound . Polymyxin B nonapeptide retains the antiendotoxin activity of polymyxin B but is much less potent . The findings suggest that these compounds block an early step in the neutrophil priming process, possibly lipopolysaccharide attachment to or insertion into the neutrophil membrane.

J Immunol, 1989 Sep 1, 143(5), 1606 - 13
Products of lipopolysaccharide-activated macrophages (tumor necrosis factor-alpha, transforming growth factor-beta) but not lipopolysaccharide modify DNA synthesis by rat trophoblast cells exhibiting the 80-kDa lipopolysaccharide-binding protein; Hunt JS et al.; Pregnancy losses from gram negative bacterial infections could be caused by direct effects of LPS on placental cells, or indirectly via LPS activation of macrophages in the uteroplacental unit . To evaluate those alternatives, LPS, LPS-activated peritoneal cells, conditioned medium from LPS-activated peritoneal cells, and some purified and recombinant molecules known to be secreted by activated macrophages were tested for their abilities to modify DNA synthesis by rat trophoblast cells . Three trophoblast cell lines derived from midgestation placentas of outbred and inbred rats were used for the experiments . Although the 80-kDa LPS-binding protein was demonstrated on trophoblast cells, LPS alone had no effect on the ability of trophoblast cells to synthesize DNA . In cocultures, trophoblast cell DNA synthesis was slightly enhanced by low concentrations of both unstimulated and LPS-activated peritoneal cells . At higher concentrations, LPS-activated cells caused significant inhibition of DNA synthesis by trophoblast cells . Conditioned media from LPS-activated peritoneal cells were highly inhibitory to trophoblast cell DNA synthesis . When specific molecules likely to be components of those media were tested, IL-1 was found to have a modest but reproducible stimulatory effect and PGE2 did not change trophoblast cell incorporation of {3H}TdR . In contrast, trophoblast cell DNA synthesis was markedly inhibited in a dose-dependent manner by both TNF-alpha and TGF-beta 1 . No differences in the sensitivity of trophoblast cells from outbred and inbred rats were observed . Given the limitations of the experimental model system, the results suggest that in cases of infection by gram-negative bacteria LPS may have an adverse effect on pregnancy by stimulating resident macrophages to generate and release molecules that are inhibitory to trophoblast cell DNA synthesis.

Med J Aust, 1989 Aug 21, 151(4), 224 - 32
Prospective audit of aminoglycoside usage in a general hospital with assessments of clinical processes and adverse clinical outcomes; Li SC et al.; A comprehensive, multiphasic review of gentamicin and tobramycin utilization was undertaken with audits of the microbiological sensitivity of Gram-negative pathogens; indications for the prescription of aminoglycoside agents; the utilization of assay services; the adequacy of clinical drug delivery by measures of serum antibiotic levels; and the assessment of adverse outcomes by markers of nephrotoxicity . The great majority of clinical isolates of target organisms (n = 4208) was more sensitive to gentamicin (96%) and to tobramycin (99%) than to all alternative agents, including first- and third-generation cephalosporin agents . A review of the indications for the prescription of aminoglycoside agents by clinical criteria showed that in 85.6% of 278 documented cases, the choice of agent was appropriate by clinical and microbiological criteria . In a substantial (77.6%) proportion of the 511 patients who were receiving therapeutic courses of an aminoglycoside agent, serum drug assays had been performed . Assay data could not be interpreted adequately in 52.6% of 3079 assayed cases as a result of inadequate data on administration regimens (39.7%) or sampling regimens (12.9%) . Where sampling was documented adequately, there was extreme variation (zero to five hours) in post-dose sampling . In only 33.2% of cases could it be concluded unambiguously that the patients were receiving safer, adequate therapy for clinically significant infections, 5.6% of patients were receiving potentially toxic doses, and 8.6% of patients showed suboptimal concentration profiles . The majority of potentially toxic levels were associated with adverse effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Z Hautkr, 1989 Aug 15, 64(8), 634 - 6, 639-43
{Skin and genital infections caused by gram-negative bacteria}; Mensing H; We discuss the classification of gram-negative bacteria as well as the most common dermatological disorders and reactions due to these germs . In addition, we give some information both on the bacteriological identification and the therapeutical concepts in these dermatoses.

Hosp Pharm, 1989 Sep, 24(9), 701 - 4
The extended-spectrum penicillins: microbiologic, utilization, and cost review in a community hospital; Stein GE et al.; The extended-spectrum penicillins have a similar spectrum of activity and clinical efficacy . The newer acylaminopenicillins (azlocillin, mezlocillin, piperacillin) offer several potential therapeutic advantages over the carboxypenicillins (carbenicillin, ticarcillin), but they are more expensive per gram . Literature reviews of these penicillins suggest that individual hospital susceptibility patterns and cost should guide selection of the most appropriate antibiotic for formulary addition . Consequently, the authors performed a microbiologic, utilization, and cost review of these antibiotics in a community hospital . From this hospital and literature review, the authors were able to make the following conclusions: (1) the newer acylaminopenicillins exhibited similar activity against common clinical pathogens and were microbiologically more active than the carboxypenicillins; (2) none of these antibiotics used alone appears adequate for empiric treatment of serious systemic gram-negative bacillary infections; (3) the extended-spectrum penicillins were prescribed in a similar fashion and usually in conjunction with an aminoglycoside; and (4) significant cost savings can be realized when less expensive drugs are used . The authors found only minor differences in microbiologic activity and clinical use among the newer extended-spectrum penicillins . Therefore, they recommend the least expensive drug for routine use in the hospital.

J Chromatogr, 1989 Aug 4, 476, 175 - 85
Efficient endotoxin removal with a new sanitizable affinity column: Affi-Prep Polymyxin; Talmadge KW et al.; A new affinity column packing for removal of endotoxins has been prepared by coupling USP drug-quality polymyxin B to Affi-Prep, a novel synthetic macroporous polymer . Affi-Prep Polymyxin binds endotoxins from a number of different strains of gram-negative bacteria . Endotoxin binding is not significantly affected by 10 mg/ml of bovine serum albumin or human immunoglobulin G, by 1 mg/ml sodium dodecyl sulphate, or by 1 mg/ml deoxycholate . Affi-Prep Polymyxin is stable to treatment with 1.0 M sodium hydroxide, an important property for sanitizing the resin . The resin shows a high ligand stability, since no leakage of polymyxin B from the packing could be detected . Affi-Prep Polymyxin exhibited the highest endotoxin binding efficiency when compared with several commercial agarose affinity packings.

Antimicrob Agents Chemother, 1989 Aug, 33(8), 1342 - 5
Influence of indomethacin on the intrarenal uptake of gentamicin in endotoxemic rats; Bergeron MG et al.; Gentamicin is a commonly used antibiotic for the treatment of gram-negative-bacterial infections . Bacterial endotoxin is liberated during antibiotic therapy, and we have shown that endotoxemic animals accumulate more aminoglycosides in their renal parenchyma than normal animals . Vasoactive mediators, such as prostaglandins and thromboxanes, are released after endotoxin and are involved in inflammation . Indomethacin, a nonsteroidal anti-inflammatory drug known to inhibit the synthesis of these hormones, was infused intravenously as a bolus (3.0 mg/kg) or as a bolus followed by a continuous infusion (0.75 mg/kg per h) to rats given gentamicin . Levels of gentamicin in serum and kidney were increased 2 h post-antibiotic treatment in the endotoxemic animals . Renal function was not significantly disturbed . Indomethacin given as a bolus failed to correct the disturbed intrarenal pharmacokinetics of gentamicin induced by endotoxin . However, a bolus followed by continuous infusion of indomethacin resulted in low cortical and high papillary levels of antibiotic . These changes were correlated with the inhibition of prostaglandin synthesis from the kidney . These observations suggest an important role for prostaglandins in the interaction among endotoxin, aminoglycosides, and the kidney . Specific inhibitors of arachidonic acid metabolites should be investigated to further understand the mechanisms of this interaction.

Eur J Haematol, 1989 Aug, 43(2), 112 - 9
An investigation into the effects of bacterial lipopolysaccharide on human platelets; Whitworth NH et al.; The addition of lipopolysaccharide (LPS) from Gram-negative bacteria to whole blood induced a consistent enhancement of platelet impedance aggregation (WB-PIA) and an increase in median platelet volume (MPV) . In contrast, the addition of LPS to platelet rich plasma (PRP) resulted in inconsistent changes in turbidometric platelet aggregation (TPA) and no significant change in MPV . The LPS-induced increase of MPV in whole blood was inhibited by verapamil, a calcium channel blocker . We conclude that: (a) LPS consistently activates platelets in whole blood but not in PRP; (b) LPS-induced activation of platelets is probably mediated by products released by leucocytes and/or erythrocytes; (c) the increase in MPV induced by LPS is probably dependent upon calcium influx; and (4) an increase in MPV may be a useful and sensitive model for the study of platelet activation by LPS, in vitro and in vivo.

Am J Physiol, 1989 Aug, 257(2 Pt 1), L65 - 70
Endotoxin stimulates platelet-derived growth factor production from cultured human pulmonary endothelial cells; Albelda SM et al.; The interaction of Gram-negative bacterial cell wall products (endotoxins) with endothelial cells is thought to be responsible for many of the damaging manifestations of Gram-negative sepsis . Because cultured human endothelial cells are relatively resistant to the direct cytotoxic actions of endotoxin, it is possible that many of the systemic effects of endotoxin may be caused by stimulation of endothelial cells to produce biologically active mediators which could then act on targets such as smooth muscle cells, fibroblasts, and leukocytes . We hypothesized that one such endothelial cell-derived mediator could be platelet-derived growth factor (PDGF), a protein that causes proliferation of mesenchymal cells, chemotaxis of leukocytes, fibroblasts and smooth muscle cells, and vasoconstriction . We therefore examined the effect of endotoxin on PDGF-like protein production by cultured adult human pulmonary artery endothelial cells . Twenty-four hours of endotoxin exposure resulted in a threefold increase in the steady-state levels of mRNA coding for PDGF B-chain (c-sis) and a two- to threefold increase in the amount of newly synthesized PDGF released into the media, as measured by immunoprecipitation of {35S}methionine-labeled protein with anti-PDGF antiserum . We conclude that human pulmonary artery endothelial cells in culture are stimulated both to produce increased amounts of PDGF mRNA and to release PDGF-like protein after exposure to endotoxin . This increased release of PDGF-like protein by human endothelial cells may play a role in the inflammatory infiltrate, vasospasm, and fibroblast proliferation that characterize the host response to endotoxin.

J Antimicrob Chemother, 1989 Aug, 24(2), 111 - 9
The type VII dihydrofolate reductase: a novel plasmid-encoded trimethoprim-resistant enzyme from gram-negative bacteria isolated in Britain; Amyes SG et al.; Plasmid pUN835 was identified in an Escherichia coli strain isolated from an outbreak of porcine diarrhoea on a farm near Nottingham, UK . The trimethoprim resistance gene did not hybridize with any of the gene probes derived from known plasmid-encoded trimethoprim resistance genes . The trimethoprim resistance gene of pUN835 was shown to encode the production of a dihydrofolate reductase which confers high-level resistance on its host . This enzyme was smaller than most plasmid-encoded dihydrofolate reductases (molecular mass = 11,500) and was labile to heat . It had relatively low affinity for the substrate dihydrofolate (Km = 20 microM) and it was resistant to competitive inhibition by trimethoprim (Ki = 7.0 microM) . We classify this novel enzyme as type VII.

Gene, 1989 Aug 1, 80(1), 161 - 9
New derivatives of transposon Tn5 suitable for mobilization of replicons, generation of operon fusions and induction of genes in gram-negative bacteria; Simon R et al.; Three types of new variants of the broad-host-range transposon Tn5 are described . (i) Tn5-mob derivatives with the new selective resistance (R) markers GmR, SpR and TcR facilitate the efficient mobilization of replicons within a wide range of Gram-negative bacteria . (ii) Promoter probe transposons carry the promoterless reporter genes lacZ, nptII, or luc, and NmR, GmR or TcR as selective markers . These transposons can be used to generate transcriptional fusions upon insertion, thus facilitating accurate determinations of gene expression . (iii) Tn5-P-out derivatives carry the npt- or tac-promoter reading out from the transposon, and TcR, NmR or GmR genes . These variants allow the constitutive expression of downstream genes . The new Tn5 variants are available on mobilizable Escherichia coli vectors suitable as suicidal carriers for transposon mutagenesis of non-E . coli recipients and some on a phage lambda mutant to be used for transposon mutagenesis in E . coli.

Zentralbl Veterinarmed A, 1989 Aug, 36(7), 539 - 48
Modulation of gap junctions in the myometrium of pigs during early gestation, induced abortion and the oestrous cycle; Rodriguez-Martinez H et al.; Myometrial samples were biopsied from pigs at early pregnancy, within 12 h after initiated abortion and at first dioestrus post-abortion . Abortion was induced either with an isolated gram-negative bacterial endotoxin or the prostaglandin (PG) F2 alpha analog cloprostenol . The tissues were quantitatively examined for gap junctions in electron micrographs . Very small numbers of gap junctions were present between smooth muscle cells in myometria at 24-27 days of pregnancy . The cell-to-cell junctions increased both in number and size at abortion, irrespective of the agent used as abortifacient . A low number of gap junctions were recorded in samples obtained after reinitiation of the oestrous cycle . The plasma levels of oestradiol-17 beta were low at the three biopsy procedures while the increased numbers of gap junctions at abortion occurred when significantly high levels of PGF2 alpha concomitantly with withdrawal of progesterone were determined in circulating blood plasma . It remains therefore unclear, which role do oestrogens play in the synthesis of gap junctions at induced abortion.

J Biol Chem, 1989 Jul 25, 264(21), 12119 - 21
O-antigen from Bradyrhizobium japonicum lipopolysaccharide inhibits intercellular (symplast) communication between soybean (Glycine max) cells; Gharyal PK et al.; The technique of fluorescence redistribution after photobleaching was utilized to measure intercellular movement of low molecular weight fluorescent hydrophilic substances across the cell wall/membrane interface between contiguous soybean (Glycine max (L.) Merr . cv . Mandarin) root cells (SB-1 cell line) in tissue culture . Lipopolysaccharide (LPS) purified from Bradyrhizobium japonicum R110d, a Gram-negative bacterium that normally infects and induces nodulation in soybean roots in vivo, inhibits intercellular communication between the soybean cells in a dose-dependent manner . In contrast, LPS from noninfecting strains failed to yield the same effect . The inhibitory activity of the LPS was localized to the O-antigen region of the LPS.

J Biol Chem, 1989 Jul 15, 264(20), 11663 - 70
Topology of the transposon Tn10-encoded tetracycline resistance protein within the inner membrane of Escherichia coli; Eckert B et al.; The transposon Tn10-encoded tetracycline resistance protein TetA is an integral membrane protein responsible for the export of tetracycline from the cytoplasmic to the periplasmic side of the inner membrane of Gram-negative bacteria . From a plot of the average hydrophobicity along the sequence of this protein, a two-dimensional membrane topology with 12 transmembrane domains may be predicted . Using plasmid-bearing Escherichia coli maxicells we specifically radiolabeled the TetA protein . The amino terminus of this membrane protein was shown not to be processed, and its location on the inner side of the cytoplasmic membrane was demonstrated by a newly developed use of a chemical method . Spheroplasts and inside-out vesicles of the TetA protein synthesizing maxicells were subjected to limited digestion by proteases of different specificities . The TetA protein was not accessible to proteases from the periplasmic side . On the inner side of the cytoplasmic membrane, the carboxyl terminus and four sites accessible to endoproteases could be identified . The cleavage sites are proposed to be localized between amino acid residues 60-70, 110-130, 180-200, and at amino acid 327 . These results allow the definition of a model for the two-dimensional topology of the TetA protein.

Invest New Drugs, 1989 Jul, 7(2-3), 203 - 7
Phase I study of vincristine and escalating doses of etoposide; Jackson DV Jr et al.; A phase I trial of vincristine and etoposide was designed following the identification of a potentially synergistic antitumor effect in a murine model . The dose of vincristine was fixed (0.5 mg daily for 3 days) . Etoposide was given at 1 of 3 total dose levels (250, 500, or 750 mg/m2) per treatment . Each dose was given in 3 equal fractions and each fraction was given daily for 3 days, i.e., 83.3 mg/m2/d x 3d, 166.7 mg/m2/d x 3d, or 250 mg/m2/d x 3d . A total of 31 patients were entered into study including 10, 18, and 3 patients treated at the 250, 500, and 750 mg/m2 dose levels, respectively . Dose-limiting toxicity occurred at the 750 mg/m2 level, in which Grade 4 myelosuppression developed in all of the patients . Life-threatening gram negative sepsis occurred in two of these patients and both required platelet transfusions . Grade 3-4 WBC toxicity was observed in 9 of 16 (56%) evaluable patients treated at the 500 mg/m2 level, but reversal of toxicity was generally rapid with repeat courses given at 3 week intervals in most patients . Non-hematologic toxicity was negligible . Objective responses were observed in 2 of 4 patients with Hodgkin's disease . The starting dose of etoposide recommended for phase II trials of this agent in combination with vincristine is 500 mg/m2; dose escalation may be possible in some patients.

J Wildl Dis, 1989 Jul, 25(3), 313 - 8
The morphological features of Aegyptianella bacterifera: an intraerythrocytic rickettsia of frogs from Corsica; Desser SS et al.; Study of ultrathin sections and freeze-fracture replicas of erythrocytes containing Aegyptianella bacterifera (Rickettsiales; Anaplasmataceae) revealed that the organisms develop within a membrane bound vacuole in the erythrocyte cytoplasm . The organisms divide by binary fission to produce about 12 rickettsiae in a mature inclusion . The organisms have gram-negative cell envelopes . The distribution of intramembranous particles within the outer and plasma membranes of A . bacterifera is similar to that of other rickettsiae and gram-negative bacteria such as Escherichia coli . The definition of the genus Aegyptianella should be broadened to include rickettsiae measuring up to 5 microns in length prior to division which develop within membrane bound inclusions in erythrocytes of their hosts.

J Clin Microbiol, 1989 Jul, 27(7), 1671 - 2
Rahnella aquatilis, an unusual gram-negative rod isolated from the bronchial washing of a patient with acquired immunodeficiency syndrome; Harrell LJ et al.; Rahnella aquatilis, a rare enteric gram-negative rod which is usually found in fresh water, was isolated from the bronchial washing of a patient with acquired immunodeficiency syndrome . Although few clinical isolates have been reported, this is the second isolation of R . aquatilis from a human in North Carolina . A case report and discussion of R . aquatilis is presented.

Circ Shock, 1989 Jul, 28(3), 249 - 55
Alterations in Gc levels and complexing in septic shock; Lee WM et al.; Septic shock involves increased generation of eicosanoids from arachidonic acid . Gc (vitamin D-binding protein) has been recently found to bind the parent molecule arachidonic acid but can also complex actin released as a result of tissue damage which causes displacement of bound arachidonic acid . Possible changes in serum levels of Gc and extent of complexing were therefore investigated in patients with gram-negative sepsis . As compared to healthy controls, serum levels of Gc were significantly decreased in patients with septic shock (P less than 0.01) . Moreover, the percentages of Gc circulating in complexed form were significantly increased (P less than 0.01) and correlated strongly with disease severity, with levels often greater than 90% in patients who died (normal mean 8% +/- 3) . These results suggest a hitherto unsuspected role for Gc in septic shock syndrome.

Allergy, 1989 Jul, 44(5), 336 - 41
Occupationally related respiratory symptoms in trout-processing workers; Sherson D et al.; All eight production workers from a small trout-processing factory were studied because of respiratory or rhinitis symptoms when working next to the automatic gutting machine . Seven of the eight had positive histamine provocation tests . Four patients had elevated total IgE levels while one had a slightly elevated eosinophil count . All workers had either weakly or moderately positive RAST against contaminated water from the outlet drain of the gutting machine . This water contained 1 microgram endotoxin/ml . We concluded that five of the workers had probably developed occupationally related asthma from inhalation of contaminated aerosol from the gutting machine . Inhalation of endotoxin from gram-negative bacteria may be the cause of this disorder . An alternative explanation is a Type I allergy caused by fish or bacterial protein with or without a simultaneous effect of endotoxin.

Pneumonol Pol, 1989 Jul-Sep, 57(7-9), 403 - 8
{CEFOBID in the treatment of respiratory tract infection (preliminary report)}; Filipecki S et al.; The therapeutical efficacy of CEFOBID (Pfizer) a third generation cephalosporine (natrium cephaperasone salt) was studied in 10 patients with acute respiratory infections including 6 with pneumonias . The therapeutical effect was arbitrarily judged according to the applied three point scale--"good", "satisfactory", "lack of effect" . A "good" effects was seen in 6 patients, in 2 "satisfactory", in further two "lack" of effect was observed . Thus CEFOBID proved to be of beneficial effect in eight out of ten patients . One should stress that in the group that responded to treatment were all cases of pneumonia . CEFOBID was well tolerated . In this preliminary study the beneficial therapeutical effect in Gram negative opportunistic infections is stressed.

Diagn Microbiol Infect Dis, 1989 Jul-Aug, 12(4 Suppl), 131S - 137S
Induction of beta-lactamase in Gram-negative bacteria; Wiedemann B et al.; The induction of beta-lactamase in Gram-negative bacteria in vitro has been established . It is possible to distinguish between high and low beta-lactamase inducers in vitro . This differentiation is clinically irrelevant because induction has little effect on the treatment of a bacterial infection with beta-lactam antibiotics . Regardless of the amount of induced beta-lactamase, the kill kinetics are usually not affected . In mutated cells, the regulatory mechanism is destroyed by inactivation of the relevant genes with respect to their regulatory function, particularly by inactivation of the amp D gene . These mutants overproduce the beta-lactamase constitutively, which results in an enzyme level that significantly exceeds the induced level . The induction process is probably not the cause of clinical failures associated with the use of beta-lactam antibiotics . It is concluded that the selection of resistant mutants with constitutive overproduction of beta-lactamase is the reason for most of these treatment failures.

Z Rheumatol, 1989 Jul-Aug, 48(4), 200 - 3
{Detection of endotoxin in synovial tissue of patients with inflammatory-rheumatic diseases}; Wagener P et al.; The etiology of inflammatory rheumatic diseases is unknown . The hypothesis, that bacteriae may play a part in the induction of a rheumatic synovialitis is further substantiated by the detection of Endotoxin (Lipid A) in the synovial membranes of five of eleven patients with early synovitis (disease duration shorter than 6 months) by using anti Lipid A monoclonal antibodies . Endotoxin (Lipid A) is a component of gram-negative bacteriae and Chlamydiae . It is a potent immunogen, and could be the link between infection of the gut or the urogenital system and the rheumatic synovialitis.

Am J Vet Res, 1989 Jul, 50(7), 1138 - 40
Rapid decay of serum IgG recognizing gram-negative cell wall core antigens in neonatal calves; Douglas VL et al.; Serum immunoglobulins of the IgG isotype recognizing common gram-negative cell core epitopes were serially measured, using a direct ELISA, on samples obtained from 20 neonatal Holstein calves . An R-mutant Escherichia coli (strain J5) was used as a plate antigen in this assay . Total serum IgG concentration was measured using radial immunodiffusion . Half-lives of core antigen-specific IgG (7.56 days) and total serum IgG (22.66 days) were dramatically different (P less than 0.0005) . This may be an indication of cross-reactive consumption of core antigen-specific immunoglobulins.

Noseleutike, 1989 Jul-Sep, 28(129), 176 - 85
{Infections in patients with cancer, criteria for quality of nursing care}; Apostolopoulov E; In this paper the factors that lead to nosocomial infections of hospitalized patients with cancer and prophylaxis measures are discussed . Infection is the primary cause of death for the individual with cancer . The typical signs and symptoms often are absent in these patients . Although Gram negative organisms account for the majority of infections today, an increase of nonbacterial infections is noted . "Good" nursing care of these patients is the major factor in preventing and minimizing the incidence of nosocomial infections and it is an indicator of quality care.

J Burn Care Rehabil, 1989 Jul-Aug, 10(4), 327 - 30
Control of endotoxemia in burn patients by use of polymyxin B; Munster AM et al.; A group of patients with severe burns were entered into two sequential prospective randomized trials for reduction of endotoxemia by the use of intravenous polymyxin B . The first group underwent polymyxin administration during the first week after burn injury in a bell-shaped dosage form constructed to resemble the level of endotoxemia as previously documented . This group showed a statistically highly significant reduction in endotoxin levels and a suggestive, but not statistically significant, reduction in wound infection and mortality in the treated group compared with controls . The second group of patients underwent treatment with perioperative polymyxin B given in conjunction with an excisional procedure of the burn wound . In this group, polymyxin B also accomplished a reduction in endotoxemia from preoperative to postoperative cases, but there was no significant reduction in clinical complication rate or mortality . In the dosages used, polymyxin B is nontoxic and promises to be a useful part of the surgeon's armamentarium in dealing with severe complications of gram-negative sepsis.

Infect Immun, 1989 Jul, 57(7), 2086 - 91
Macrophage activation by an ornithine-containing lipid or a serine-containing lipid; Kawai Y et al.; alpha-N-(3-Acyloxyacyl)-ornithine (or -serine) is the structure of lipoamino acids obtained by us previously from some gram-negative bacteria (Y . Kawai and I . Yano, Eur . J . Biochem . 136:531-538, 1983; Y . Kawai, I . Yano, and K . Kaneda, Eur . J . Biochem . 171:73-80, 1988; Y . Kawai, I . Yano, K . Kaneda, and E . Yabuuchi, Eur . J . Biochem . 175:633-641, 1988) . The 3-acyloxyacylamide structure is present in both the lipoamino acids and lipid A of lipopolysaccharide (endotoxin) . The efficacy of lipoamino acids (an ornithine-containing lipid and a serine-containing lipid) in activating C3H/HeSlc mouse peritoneal exudate macrophages was compared with that of bacterial lipopolysaccharide, because the two types of substances were expected to exhibit similar biological activities and physiological functions on the basis of their structural similarities . Actually, the lipoamino acids, as well as lipopolysaccharide, strongly activated the macrophages to generate the immunoregulatory substances prostaglandin E2 and interleukin-1, but their effect on the induction of L929 cell cytolytic factor (a possible tumor necrosis factor), another immunoregulatory substance, was weaker than that of lipopolysaccharide . The effect of lipoamino acids on the cytotoxicity of macrophages for EL-4 leukemia cells was very weak . However, all of these activities, as far as tested, were strongly enhanced by synergistic action with gamma interferon . Only the serine-containing lipid killed both C3H/HeSlc and C3H/HeJ macrophages to almost the same degree as endotoxin killed C3H/HeSlc macrophages . On the other hand, lethal toxicity for mice was not found with either the ornithine-containing lipid or the serine-containing lipid, even when 7 mg of compound was injected into a mouse . These studies suggest that the lipoamino acids are nontoxic characteristic immunoactivators.

J Biol Chem, 1989 Jun 25, 264(18), 10867 - 71
Identification of a lipid A binding site in the acute phase reactant lipopolysaccharide binding protein; Tobias PS et al.; Lipopolysaccharide (LPS) binding protein (LBP), a recently discovered 60-kDa acute phase protein, is present in the acute phase serum of many species including human, rabbits, mice, and rats . Using either highly purified LBP from acute phase rabbit serum or unfractionated acute phase rabbit serum as a source of LBP, we examined the binding of LBP to LPS immobilized on plastic microtiter plates and to LPS electrotransferred to nitrocellulose after sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The presence of LBP bound to LPS was detected with goat anti-rabbit LBP and peroxidase-conjugated rabbit anti-goat IgG . LBP was found to bind to a variety of LPS types from both rough and smooth strains of Gram-negative bacteria, to lipid A, and to the tetraacyl glucosamine disaccharide diphosphate precursor IVA, but bound very poorly to the diacyl glucosamine phosphate, lipid X . No binding to 3-deoxyoctulosonic acid was observed . Binding affinities for LPS are near 10(9) M-1 . The data presented here support the concept that LBP contains a binding site for lipid A.

Med J Aust, 1989 Jun 19, 150(12), 720 - 1
Near-fatal septicaemia with Chromobacterium violaceum; Georghiou PR et al.; Chromobacterium violaceum is a Gram-negative organism which normally inhabits water and soil . Human infection is unusual and is associated with a high mortality rate . We describe a typical case of disseminated infection with Chr . violaceum in a male carpet cleaner . The possible origin and treatment of the infection is discussed.

Am J Med, 1989 Jun 9, 86(6A), 85 - 93
Side effects of stress bleeding prophylaxis; Tryba M; Conventional stress bleeding prophylaxis with antacids or histamine (H2)-antagonists, as well as the newer mucosa-protective drugs pirenzepine and sucralfate, are satisfying most of the clinicians with regard to efficacy of stress bleeding prevention . Therefore, potential side effects are attaining crucial importance with regard to the drugs to be used . Pharmacologic blockade of cardiac H2-receptors increases the risk of bradycardia and negative inotropic effects as well as coronary vasoconstriction at least in the presence of elevated plasma histamine levels . Intracardiac injection of pirenzepine can lead to temporary tachycardia . Elderly patients have been shown to be at an increased risk of side effects to the central nervous system when treated with H2-antagonists . These drugs can also induce toxic effects in the liver . Cimetidine leads to interactions with a number of drugs used in the intensive care unit . In patients with pre-existing pulmonary diseases, H2-antagonists have been demonstrated to increase pulmonary bronchoconstriction . Alkalinization of the gastric juice is associated with a significant increase in colonization of gram-negative bacteria in the stomach . In intubated patients, aspiration of stomach contents occurs in 30 to 40 percent of the patients . A number of studies have shown a direct correlation between alkalinization of the gastric juice and pulmonary infections . Sucralfate and to a lesser degree pirenzepine can reduce the risk of pulmonary infections . Sucralfate also exerts a bactericidal effect . Recent investigations support the hypothesis that alkalinization of the stomach also increases the risk of systemic infections . This may be the main reason for the observation that at least in ventilated patients sucralfate, unlike H2-antagonists or antacids, leads to a significant reduction of the mortality rate compared with conventional stress bleeding prophylaxis.

Zh Mikrobiol Epidemiol Immunobiol, 1989 Jun, (6), 91 - 4
{Effect of inducers and inhibitors of mixed function oxidases on body resistance to endotoxins of gram-negative bacteria}; Liniuchev MN et al.; Experimental typhoid intoxication in white mice leads to the inhibition of microsomal oxidation in the liver, which is manifested by the prolongation of hexenal-induced sleep and a decrease in the toxic action of parathion . Phenobarbital, capable of inducing oxidases with mixed function (OMF), enhances the process of the detoxification of endotoxin injected into the animals, which is manifested by the increase of its LD50 . Soluble levomycetin succinate, widely used for the treatment of typhoid-paratyphoid infections, is a powerful inhibitor of OMF (as shown by the hexenal test) . Benzonal, the analog of phenobarbital, removes the inhibitory effect of the antibiotic . Experimental studies carried out in the course of this investigation make it possible to substantiate the clinical trial of these preparations (OMF inducers) used in the complex therapy of typhoid-paratyphoid infections for the stimulation of natural detoxification mechanisms of the body . Benzonal is the preparation of choice for use in clinical practice.

Pathol Biol (Paris), 1989 Jun, 37(5 Pt 2), 643 - 48
{Effects of antibiotics on the production of prostaglandins by macrophages in vitro}; Garraffo R et al.; Many reports have pointed out a relationship between antibiotics and immunity . It has been shown that antibiotics are able to modify polynuclear chemotactism, bacteria phagocytosis by macrophages, lymphocyte proliferation and the production of various cytokines (interleukins I, II...) . On the other hand, the immunoregulatory activity of metabolites of arachidonic acid is proposed to explain, in part, these properties . For example it is well established that PGE2 has a negative retro control on Interleukin production by monocytes . Moreover eicosanoids are probably involved in the inflammatory and vasoactive reactions during the septic shock syndrome observed frequently during phagocytosis and bacterial lysis of Gram negative bacteria . Thus it seems interesting to evaluate the antibiotics activity on eicosanoid monocyte production . We have studied the influence of ciprofloxacin, ofloxacin, pefloxacin, imipenem, doxycycline, clindamycin and cefadroxil at various concentrations on eicosanoid production by A 13187 calcium ionophore activated mouse peritoneal macrophages (P 388 D1) . The analyzed prostaglandins are: 6 keto (PGI2), PGE1, PGE2, F2 alpha, A2, B2, TxB2 et PG D2 . All the antibiotics tested show either an inhibiting or a stimulating action on eicosanoid production . These immunomodulatory and pro-inflammatory effects must be considered, together with anti-infectious activity before the prescription of an antibiotic.

Oral Microbiol Immunol, 1989 Jun, 4(2), 98 - 105
Opsonic IgG antibody against Actinobacillus actinomycetemcomitans in localized juvenile periodontitis; Baker PJ et al.; Actinobacillus actinomycetemcomitans is a gram-negative bacterium frequently recovered from periodontal lesions of patients with localized juvenile periodontitis (LJP) . Elevated levels of serum IgG and IgM antibodies to A . actinomycetemcomitans antigens are frequently observed in LJP patients, although the functional properties of such antibodies have not been characterized systematically . In this study, we analyzed serum from LJP subjects infected with A . actinomycetemcomitans with respect to the presence of IgG antibodies expressing opsonic, bactericidal and/or leukotoxin-neutralizing activity against this organism . The IgG fractions obtained from serum of 3 LJP patients with elevated antibody titers to A . actinomycetemcomitans contained opsonic activity against a non-leukotoxic Y4 strain, as well as for a highly leukotoxic JP2 strain . Opsonic activity required the presence of complement . The IgG fractions of pooled normal serum and serum from a fourth LJP subject with minimal ELISA-reactive IgG antibody against this organism lacked detectable opsonic activity . Leukotoxin-neutralizing IgG antibodies, although variably present, did not influence neutrophil killing of the leukotoxic JP2 strain . None of the sera tested contained bactericidal IgG antibodies capable of promoting direct complement-mediated killing of A . actinomycetemcomitans . These results indicate that LJP subjects infected with A . actinomycetemcomitans are capable of producing opsonic IgG antibodies which may facilitate neutrophil-mediated host defense against this periodontopathic organism.

Arch Intern Med, 1989 Jun, 149(6), 1449 - 51
A pink-pigmented, oxidative, nonmotile bacterium as a cause of opportunistic infections; Korvick JA et al.; We describe two cases of bacteremia due to a pink-pigmented, oxidative, nonmotile, gram-negative, rod-shaped organism . One case occurred in a febrile neutropenic patient and another in a chronically debilitated patient with pancreatic abscess . The first patient was cured with gentamicin and ticarcillin, but the second patient died while receiving cefamandole therapy . The organisms described here are similar to Methylobacterium mesophilicum (Pseudomonas mesophilica) and the "unnamed taxon" organisms . A major difference from M mesophilicum is the lack of methanol utilization . Further distinctions between our isolates and M mesophilicum are the lack of flagella in our organisms, growth at 42 degrees C, growth on MacConkey's agar, lack of acetamide assimilation, and citrate utilization . The lack of flagella is the principle difference between our isolates and those in the unnamed taxon . Both of the isolates were resistant to the cephalosporins, but susceptible to the aminoglycosides, ticarcillin-clavulanic acid, sulfamethoxazole and trimethoprim, and imipenem . With the growing population of immunocompromised and chronically ill patients, these organisms may emerge as important pathogens.

Arch Intern Med, 1989 Jun, 149(6), 1314 - 6
Pefloxacin treatment of meningitis caused by gram-negative bacteria; Segev S et al.; Ten patients with acute meningitis caused by gram-negative bacteria were treated with pefloxacin intravenously for a mean period of 10 days . Eight patients responded clinically to pefloxacin treatment, and the causative organisms were eradicated from the cerebrospinal fluid in 9 of the 10 patients . Pefloxacin may offer a new, efficacious, and safe therapy for gram-negative meningitis.

Gastroenterology, 1989 Jun, 96(6), 1589 - 95
Preserved cytosolic and synthetic liver function in jaundice of severe extrahepatic infection; Pirovino M et al.; We investigated prospectively 9 adult patients with the syndrome of jaundice complicating severe extrahepatic infection both clinically and by quantitative liver function tests . Five patients having severe extrahepatic infection without jaundice were used for comparison . Intraperitoneal infection was found to be a major risk factor for development of jaundice . Jaundice was mainly associated with gram-negative infection, but did not influence survival . Duration of jaundice was dependent on control of the underlying infection . Liver function tests showed a severely deranged organic anion transport, whereas synthetic, cytosolic, and microsomal functions remained preserved . Our study shows that (a) the syndrome of jaundice associated with extrahepatic infection is a functional disorder that is reversible upon control of infection, and that (b) cytosolic, synthetic, and microsomal function is preserved . This may have consequences for both assessing prognosis and clinical management.

J Chemother, 1989 Jun, 1(3), 162 - 3
Serum sensitivity of Escherichia coli strains present in the clinical environment; Miglioli PA et al.; It is well known that the growth of some Gram-negative microorganisms is inhibited by normal human serum; but there are no reports about the percentage of serum-sensitive Escherichia coli strains present in clinics . The aim of the present paper is to obtain data useful for elucidating this point . The study was carried out using 38 strains of E . coli isolated in clinics . Different concentrations of serum (from 2.5% to 20% in BHI broth) were used for evaluating its bactericidal activity . The bactericidal activity was measured both as colony forming units (CFU) and as a delay in the growth curve recorded by an Avantage-Abbott instrument . The concentration of serum inducing a killing effect greater than 99.9% was considered bactericidal . The results indicate that in the presence of 2.5%, 5%, 10% and 20% of serum in BHI broth, the percentage of serum-sensitive strains was 2.6%, 10.5%, 18.4% and 31.6%, respectively . These data are discussed in view of the possible application of the synergistic effect of serum with sub-MICs of different antibiotics on serum-sensitive E . coli strains present in the clinical environment.

Obstet Gynecol Clin North Am, 1989 Jun, 16(2), 291 - 304
Cephalosporins; Martens MG; The cephalosporins have had a great effect on the prevention and treatment of many obstetric and gynecologic infections . Broader spectra and greater activity often allow for effective single-agent usage . Improved pharmacokinetics have permitted the number of doses to be decreased . Excellent activity against several gram-negative organisms sometimes permits substitution of the safer cephalosporins for the aminoglycosides, thus avoiding potential renal and otic toxicities . However, these advantages come with costs of their own, such as superinfection, induction of resistance, and the potential for clinical bleeding . Nevertheless, these adverse effects fortunately are infrequent and, with careful monitoring, they can be readily avoided or controlled . Therefore, the increasing number and diversity of these helpful agents should be accompanied by greater knowledge of all these factors . This should result in increasing benefits for our patients, without sacrificing cost, safety, and future therapeutic success.

Surg Gynecol Obstet, 1989 Jun, 168(6), 481 - 90
Use of topical antiseptic in prophylaxis of catheter-related septic complications; Illner H et al.; Pretreatment of catheters by a simple new procedure designed to reduce the incidence of septic complications was tested in both in vitro and in vivo studies . In the former experiments, the high sensitivity of gram-negative bacteria and fungi to Silastic (silicone rubber) catheters pretreated with silver nitrate solution was determined . The antimicrobrial activity remained unchanged after both sterilization and up to six weeks of storage . Furthermore, prolonged incubation of treated catheter segments in Escherichia coli inoculated plasma resulted in a significant reduction of organisms in the media and those adherent to the surface of the catheters . The in vivo experiments were performed upon two groups of rabbits . In the first group, catheters (one treated and one untreated) were implanted into contralateral jugular veins . In the second group, alternating treated and control catheter segments were threaded onto polyethylene core tubing; the resulting string of segments was positioned in the inferior vena cava . Five to 11 days after implantation of the catheter, all rabbits were intravenously injected with live E . coli (approximately 10(8) to 10(9) organisms per kilogram of weight); 18 to 24 hours later, the catheters and blood samples were removed for cultivation . Significant reductions in both incidence and magnitude of colonization in treated catheters by E . coli were observed in both rabbit groups . Additionally, histologic examination did not reveal any significant differences between contralateral jugular veins (previously in prolonged and intimate contact with the indwelling catheters), confirming the absence of any local adverse effects of silver nitrate.

Gastroenterol Jpn, 1989 Jun, 24(3), 262 - 9
Non-septic endotoxemia in cirrhotic patients; Yajima Y et al.; We have found that endotoxemia detected by conventional LCT (limulus colorimetric test) in patients with liver diseases could not be detected by endotoxin-specific LCT at all, and proposed that this beta-glucan like activity (BGLA) should be termed as non-septic endotoxemia, distinguishing it from septic endotoxemia seen in gram-negative sepsis . In this study, we investigated non-septic endotoxemia through the clinical course of 8 cirrhotic patients . Non-septic endotoxemia appeared at the onset of DIC but tended to decline in level in the late terminal stage . This phenomenon cannot be consistent with the "spillover" theory which explains the mechanism of endotoxemia without sepsis in liver disease . We think it is an urgent problem to elucidate the nature of BGLA in liver disease, without recourse to the "spillover" theory.

Am Rev Respir Dis, 1989 Jun, 139(6), 1361 - 8
Pentoxifylline prevents tumor necrosis factor-induced lung injury; Lilly CM et al.; Human tumor necrosis factor-alpha (TNF) is a monokine produced by mononuclear cells after many stimuli, including bacterial endotoxin . Full exploration of its antineoplastic effects has been limited by side effects . We have previously shown that the administration of TNF to guinea pigs is associated with a syndrome similar to gram-negative septic shock, which includes capillary permeability lung injury . In this study, we measured the effects of pentoxifylline (PTX) on parameters of TNF-induced lung injury including: lung wet-to-dry weight ratio, the ratio of lung-to-plasma 125I-labeled albumin (albumin index), bronchoalveolar lavage (BAL) and peripheral leukocyte counts, and serial measurements of mean arterial pressure (MAP) . Four groups of animals were studied: a TNF group received 3.75 x 10(6) U/kg TNF; a PTX group received a 20-mg/kg bolus of PTX followed by an infusion of 6 mg/kg/h; the PTX-TNF group received both; and the final group was a saline control . ANOVA analysis revealed significant elevations of lung wet-to-dry ratio only in the TNF group (5.9 {5.6 to 6.3}, p less than 0.001), expressed as the mean followed by 95% confidence intervals) . Lung albumin index was elevated only in the TNF group (0.24 {0.19 to 0.29}, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

J Exp Med, 1989 Jun 1, 169(6), 1977 - 91
Tumor necrosis factor/cachectin increases permeability of endothelial cell monolayers by a mechanism involving regulatory G proteins; Brett J et al.; Endothelium is an important target of tumor necrosis factor/cachectin (TNF), a central mediator of the host response in endotoxemia and Gram-negative sepsis . In this report, TNF is shown to increase the permeability of endothelial cell monolayers to macromolecules and lower molecular weight solutes by a mechanism involving a pertussis toxin-sensitive regulatory G protein . Within 1-3 h of exposure to TNF (5 nM), changes in cell shape/cytoskeleton occurred that led to disruption of monolayer continuity with the formation of intercellular gaps . Correlated with these structural changes was an increase in endothelial permeability to macromolecular and lower molecular weight tracers; time-dependent, reversible increases in passage of these tracers, evident by 1-3 h, were observed after addition of TNF to cultures . Perturbation of barrier function by TNF also depended on the dose of TNF added being half-maximal by approximately 0.4 nM . Only a brief exposure (15 min) of TNF to endothelium was required to induce an increase in permeability, and this was not prevented by the presence of cycloheximide or actinomycin D . Preincubation of monolayers with pertussis toxin blocked in parallel TNF-induced increased passage of solutes and cell shape/cytoskeletal perturbation, indicating the close correlation between these changes in endothelial cell function . In contrast, pertussis toxin did not alter TNF-induced modulation of two endothelial cell coagulant properties . These data provide evidence for two intracellular pathways of TNF action that are distinguishable by pertussis toxin and provide insight into a mechanism underlying loss of solute from the intravascular space mediated by TNF: alteration in endothelial cell barrier function.

Blood, 1989 Jun, 73(8), 2188 - 95
Endotoxin-induced production of plasminogen activator inhibitor by human monocytes is autonomous and can be inhibited by lipid X; Schwartz BS et al.; Peripheral blood mononuclear cells (PBMs) produce both tissue factor and plasminogen activator inhibitor type 2 (PAI-2) in response to gram-negative bacterial lipopolysaccharide (LPS) . The cellular roles in the tissue factor response have been previously elucidated, and we now report those roles in PAI-2 production . Monocytes are the only cells among LPS-stimulated PBMs that produce PAI-2 as assessed by measurement of PAI-2 activity and antigen . Concomitant immunohistochemistry demonstrated that monocytes contain PAI-2, with a greater number staining positively and more intensely after exposure to LPS . LPS-stimulated monocytes produced increased amounts of PAI-2 with or without addition of lymphocytes . Lymphocytes prestimulated with LPS and then washed did not induce PAI-2 production in monocytes to which they were added . Lipid X, a precursor in the biosynthetic pathway of lipid A and LPS, was able to inhibit LPS induction of monocyte PAI-2 in a dose-dependent manner . This inhibition was not due to cellular toxicity, the phospholipidlike nature of lipid X, interference with the PAI-2 assay, or monocyte production of a substance interfering with PAI-2 . Lipid X was an effective inhibitor of PAI-2 production even when added up to 30 minutes after LPS.

Rev Prat, 1989 May 25, 39(16), 1395 - 8
{Nosocomial bronchopneumopathies}; Durocher A et al.; Nosocomial bronchopulmonary infections are common and severe complications, particularly in intensive care units . The high incidence of pneumonia is related to multiple factors such as underlying disease, acute respiratory failure, nutritional disorders, depressed mental status and the frequent need tracheal intubation . The most frequent cause of respiratory tract infection is aspiration of oropharyngeal secretions . In hospitalized patients, there is usually an oropharyngeal colonization with Gram-negative bacteria . Prevention of nosocomial bronchopulmonary infections requires close attention to the patient's environment, proper techniques, handwashing and decontamination of respiratory equipment.

Mol Cell Biochem, 1989 May 4, 87(1), 79 - 84
Involvement of cytochrome b5 in the cytotoxic response to Escherichia coli lipopolysaccharide; Portoles MT et al.; Cytotoxic lesions, induced by Gram-negative lipopolysaccharides (LPS), occur mainly in liver where the microsomal compartment of hepatocytes is involved in the detoxification mechanisms as well as in the biosynthesis of different active metabolites . The alterations induced by LPS from E . coli 0111:B4 on cytochrome b5 and its correlation with cytochrome P450, have been studied using an in vivo reversible endotoxic shock model and 24 h non-replicative hepatocyte monolayers . Results show that cytochrome b5 is directly affected by LPS that induces also a membrane damage with an active release of lactate dehydrogenase (LDH) . The increase of cytochrome b5 levels may enhance the efficiency of the electron transport, thus facilitating the cytochrome P450-associate oxidations and reactions involved in the repair mechanisms of membranes.

Appl Environ Microbiol, 1989 May, 55(5), 1289 - 90
Growth of the syntrophic anaerobic acetogen, strain PA-1, with glucose or succinate as energy source; Brulla WJ et al.; Strain PA-1 (S . Barik, W.J . Brulla, and M.P . Bryant, Appl . Environ . Microbiol . 50:304-310, 1985) is an anaerobic, gram-negative rod that in pure culture decarboxylates succinate to propionate and that grows syntrophically as an acetogen with the H2 utilizer Methanospirillum hungatei if glucose, pyruvate, aspartate, or fumarate is provided . In pure culture, strain PA-1 grows optimally in a medium containing 5% ruminal fluid, 0.1% yeast extract, a 4:1 N2-CO2 gas phase, and 20 mM succinate . With the PA-1 plus M . hungatei coculture, good growth was obtained with 7.5 mM glucose and tryptophan could replace the yeast extract . Strain PA-1 in pure culture grew quite well in glucose medium if the large headspace was flushed intermittently with N2 . Flushing with H2 inhibited this growth.

J Dairy Sci, 1989 May, 72(5), 1137 - 41
Conventional tube and microplate Limulus amoebocyte lysate procedures for determination of gram-negative bacteria in milk; May SA et al.; A comparison was made of the conventional tube and microplate Limulus amoebocyte lysate assay for detection of gram-negative bacterial lipopolysaccharide in milk . Raw whole milk samples were maintained frozen and portions were examined periodically on alternate days during 13-d storage to evaluate the reproducibility of both Limulus amoebocyte lysate procedures and to determine optimum reaction conditions for the microplate method . One-day-old, raw and locally purchased pasteurized milk samples, held at 7 degrees C, were analyzed during storage to establish the correlation of both procedures with aerobic and modified psychrotrophic plate counts . Vitamin- and mineral-fortified dairy-based products were examined using the microplate Limulus amoebocyte lysate test as a potential indicator of raw material or finished product bacterial quality and possible postprocessing contamination . Statistical analysis of the data collected comparing the conventional tube and the microplate Limulus amoebocyte lysate assay demonstrated no significant difference exists between the methods when either the modified psychrotrophic bacterial count or the aerobic plate count was used to determine gram-negative bacteria in pasteurized or raw milk (P less than .91) . The microplate method, which uses half the lysate reagent, was a good indicator of the bacterial quality of milk and fortified dairy products, consistently detecting bacterial levels greater than 10(3) to 10(4)/ml.

Circ Shock, 1989 May, 28(1), 49 - 58
Antithrombin-III treatment limits disseminated intravascular coagulation in endotoxemia; Redens TB et al.; Gram-negative septicemia/endotoxemia remains a serious clinical disorder that is often complicated by disseminated intravascular coagulation (DIC) . Plasma antithrombin-III (AT-III) levels usually decrease during gram-negative septicemia/endotoxemia, and even moderate decreases in this major inhibitor of the coagulation system are associated with serious DIC . We demonstrated in an earlier study that prophylactic treatment of rats with 250 U/kg of AT-III followed by endotoxin challenge markedly attenuates DIC, indices of organ damage, and metabolic dysfunction . The present study was to determine whether treatment with 250 U/kg AT-III 1 hr after endotoxin challenge would be similarly efficacious . Rats treated with 250 U/kg of AT-III inactivated by human sputum elastase (ATX) served as protein controls . Blood samples for analysis were obtained 4 hr after AT-III or ATX treatment (5 hr after endotoxin challenge) . Rats in the ATX treatment group exhibited abnormalities characteristic of endotoxemia, i.e., decreased fibrinogen levels and platelet counts, increases in prothrombin time and activated partial thromboplastin time, elevated serum glutamic oxaloacetic transaminase (SGOT) and alkaline phosphatase (AKP), and hypoglycemia . Treatment with AT-III markedly and significantly (P less than .05) attenuated all of these abnormalities, although survival was not increased . This study strongly suggests that supplementation of plasma AT-III is efficacious after the development of sepsis, although not as efficacious as prophylactic treatment.

J Infect Dis, 1989 May, 159(5), 931 - 7
Netilmicin in the treatment of gram-negative bacteremia: single daily versus multiple daily dosage; Sturm AW; Seventy patients with culture-proven bacteremia with gram-negative rods were randomly treated with either a single (S) dose daily or multiple (M) doses daily of netilmicin . All bacterial strains were susceptible to the antibiotic . No differences were found with respect to efficacy . Therapy for bacteremia failed in two M patients, but bacteria persisted in 17 (10 S, 7 M) at the primary infection site . In 12 there was an anatomic or physiologic factor contributing to this persistence . One S patient showed mild nephrotoxicity, but ototoxicity was not found in any of the 35 patients who underwent serial audiography . Mild reversible rises of transaminases were found in 5 of 35 S patients and in 3 of 34 M patients . Once-daily administration of netilmicin seems to be as effective and as safe as conventional multiple daily doses.

Infection, 1989 May-Jun, 17(3), 142 - 5
Three-step empiric treatment for severely neutropenic patients with fever: ceftazidime--vancomycin--amphotericin B; Koeppler H et al.; In a prospective study 50 febrile episodes in severely neutropenic patients (neutrophils less than 500/mm3) were empirically treated with ceftazidime monotherapy . If no response was seen after 48 h, vancomycin was added . After another 72 h period, patients with persisting fever additionally received amphotericin B . In 29 episodes (58%) patients became afebrile with ceftazidime monotherapy . Another seven patients (14%) responded to the addition of vancomycin and five patients (10%) needed amphotericin B to become afebrile . A success of the study drugs without modification was seen in 40 episodes (80%), success with modification in three episodes (6%) and failure in six episodes (12%) . One patient died of myocardial infarction . No other death occurred during the two-week observation period after entering the study . Though there were two gram-negative isolates resistant to ceftazidime, these patients were successfully treated with modification . It is concluded that the response-adapted additive sequence of ceftazidime, vancomycin and amphotericin B is an effective approach towards febrile episodes in severely neutropenic patients.

Biochem Pharmacol, 1989 May 1, 38(9), 1427 - 33
Quantitative relationship between sensitivity to beta-lactam antibiotics and beta-lactamase production in gram-negative bacteria--II . Non-steady-state treatment and progress curves; Frere JM et al.; A non-steady-state model is discussed for the study of the interplay between beta-lactamase activity and outer membrane permeability with slowly hydrolysed beta-lactams . The analysis shows: (1) that the simple, steady-state model presented in the accompanying paper remains valid as long as kcat (i.e . k3 with chromosome-encoded class C beta-lactamases) is larger than 10(-3)/sec (generation time = 20 min or more); (2) that among the beta-lactam antibiotics studied here, the complete, non-steady-state model needs only be used in the case of aztreonam; (3) that the term "trapping" should be replaced by "formation of a covalent acyl-enzyme" and that such a phenomenon only contributes significantly to the resistance when penetration and hydrolysis are very slow and the periplasmic beta-lactamase concentration is very high . Aztreonam seems to be the only compound which fulfils the first two conditions.

FEMS Microbiol Lett, 1989 May, 50(1-2), 203 - 6
Construction of Hfr-like donors of the obligate methanol-oxidizing bacterium Methylobacillus flagellatum KT; Serebrijski IG et al.; The transposon-loaded plasmid pAS8-121, incapable of autonomous replication in Gram-negative bacteria of non-enteric group, was transferred to Methylobacillus flagellatum KT wild type strain MFK1 . The transconjugants arose at a frequency of 10(-7) per donor cell . The majority of the transconjugants tested exhibited the transfer of all selected chromosomal markers at rather high (10(-4)-10(-6) per donor cell) but similar frequencies . Only one of the obtained donors, designated MFK 64, was capable of mobilizing M . flagellatum KT chromosome in a polarized manner . The integrated nature of the plasmid in this and other MFK1 (pAS8-121) derivatives was supported by the results of DNA-DNA hybridization.

Thromb Haemost, 1989 Apr 25, 61(2), 294 - 7
Heparin inhibition of endotoxin-dependent Limulus amebocyte lysate coagulation; Marcum JA et al.; Addition of commercial heparin to preparations of the coagulation system obtained by hypotonic lysis of Limulus amebocytes, prior to the addition of endotoxin from gram-negative bacteria, inhibited gelation . Activation of the cell-lysate proclotting enzyme, utilizing a diluted lysate preparation and synthetic substrates, was inhibited to a comparable extent with either anticoagulantly active or inactive heparin . Other glycosaminoglycans, including heparan sulfate, dermatan sulfate, chondroitin 4-sulfate, and chondroitin 6-sulfate, did not inhibit the generation of clotting enzyme activity . Experiments conducted with preparations of proactivator(s) and proclotting enzyme, obtained by fractionation of lysate with heparin-Sepharose, revealed that inhibition of endotoxin-dependent activation of amebocyte lysate by heparin is mediated by precipitation of the proactivator(s) and can be prevented by increasing the salt concentration.

Nature, 1989 Apr 20, 338(6217), 652 - 5
Fibronectin binding mediated by a novel class of surface organelles on Escherichia coli; Olsen A et al.; Gram-negative bacteria are known to produce two types of surface organelles: flagella, which are required for motility and chemotaxis, and pili (fimbriae), which play a part in the interaction of bacteria with other bacteria and with eukaryotic host cells . Here we report a third class of E . coli surface organelles for which we propose the name curli . Curli are coiled surface structures composed of a single type of subunit, the curlin, which differs from all known pilin proteins and is synthesized in the absence of a cleavable signal peptide . Although the gene encoding this structural subunit, crl, is present and transcribed in most natural isolates of E . coli, only certain strains are able to assemble the subunit protein into curli . This assembly process occurs preferentially at growth temperatures below 37 degrees C . The ability of curli to mediate binding to fibronectin may be a virulence-associated property for wound colonization and for the colonization of fibronectin-coated surfaces.

Arch Surg, 1989 Apr, 124(4), 445 - 8
Recombinant human tumor necrosis factor produces hemodynamic changes characteristic of sepsis and endotoxemia; Schirmer WJ et al.; Tumor necrosis factor (TNF) is a macrophage-derived peptide mediator released during endotoxemia and sepsis . We examined the systemic and visceral hemodynamic response to low doses of human recombinant TNF in rats . Each animal received a 30-minute intravenous infusion of either saline solution (n = 8) or TNF (n = 8) in a dose of 0.25 mg/kg or 1.0 mg/kg . Thermodilution cardiac output, blood pressure, pulse, vascular resistance, effective hepatic blood flow (galactose clearance), and effective renal plasma flow (p-aminohippurate clearance) were determined at time = 2 hours . The 0.25-mg/kg dose had no apparent effect on systemic hemodynamics . The 1.0-mg/kg dose produced a hyperdynamic systemic circulatory response with an elevated cardiac output, tachycardia, and a diminished systemic vascular resistance . Effective hepatic blood flow was exquisitely sensitive to even the lowest dose of TNF, with a 29% reduction despite the normal cardiac output . Renal flow was unaffected by either dose . Tumor necrosis factor-induced systemic and visceral hemodynamic changes are remarkably similar to those seen in gram-negative sepsis, suggesting that TNF may occupy a proximal position in the pathogenesis of overwhelming infection.

Acta Chir Scand, 1989 Apr-May, 155(4-5), 233 - 9
Polymorphonuclear leucocyte activation after trauma: evidence for increased generation of oxygen free radicals in response to in vitro endotoxin stimulation; Rokke O et al.; The effects of moderate bone trauma on the functional status of circulating polymorphonuclear neutrophil (PMN) leucocytes were studied over a 3-day period in a porcine model . Nine pigs were submitted to femur marrow nailings and six acted as controls . The PMNs were studied in regard to production of oxygen free radicals and locomotion . The trauma was followed by leucocytosis, dominated by granulocytosis . Oxygen free radical production by PMNs, measured by chemiluminescence, was significantly greater in the traumatized animals than in the controls . Stimulation of the PMNs with low concentrations of endotoxin disclosed intergroup differences until 48 hours post-trauma . No intergroup difference in PMN locomotion were demonstrated . The results indicated post-traumatic functional changes in the circulating PMNs, suggesting increased reactivity and sensitivity for gram-negative bacteria and their endotoxins during septicaemia.

Infect Immun, 1989 Apr, 57(4), 1054 - 8
Endotoxic-lipopolysaccharide-specific binding proteins on lymphoid cells of various animal species: association with endotoxin susceptibility; Roeder DJ et al.; Endotoxic lipopolysaccharide (LPS), a common structural component of all gram-negative bacteria, is well recognized for its capacity to interact with and perturb immunologically relevant cells . Using a radioiodinated, photoactivatable LPS probe, we have recently identified an 80-kilodalton LPS-specific binding protein on murine B lymphocytes . We now have extended these studies to determine if other mammalian species, as well as representative endotoxin-resistant species (frog and chicken), have a similar LPS-binding protein . We have identified what appears to be a relatively conserved 80-kilodalton LPS-binding protein on mononuclear cells of all mammalian species tested . However, both frog and chicken leukocytes failed to show the presence of a similar LPS-binding protein . It is possible that the presence of specific LPS-binding proteins may be important for endotoxin sensitivity of most mammalian species.

Appl Environ Microbiol, 1989 Apr, 55(4), 771 - 7
Differential regulation of lambda pL and pR promoters by a cI repressor in a broad-host-range thermoregulated plasmid marker system; Winstanley C et al.; Plasmid systems with unique markers were constructed to assess the fate of recombinant DNA and genetically manipulated bacteria in soil and freshwater model environments . On such constructs the marker gene, xylE (for catechol 2,3-dioxygenase), is expressed from the lambda promoter pL or pR, each of which is controlled by the temperature-sensitive lambda repressor c1857 . Combinations of these elements were cloned into the broad-host-range plasmid pKT230 to form pLV1010 (pL-xylE), pLV1011 (pL-xylE-c1857), and pLV1013 (pR-xylE-c1857) . The recombinant plasmids were introduced into different gram-negative bacteria . The thermoregulated system of pLV1013 functioned well in a range of species, with xylE induction being readily achieved by elevation of the temperature from 28 to 37 degrees C . There was a difference in the induction of catechol 2,3-dioxygenase activity, depending on whether xylE was expressed from pL (pLV1011) or pR (pLV1013) . Our observations on testing the different systems in a number of hosts suggest that genes carried by the DNA of genetically engineered microorganisms may not be expressed in a predictable manner following transfer from the release host to other species.

Appl Environ Microbiol, 1989 Apr, 55(4), 1029 - 32
Isolation and characterization of quinoline-degrading bacteria from subsurface sediments; Brockman FJ et al.; Two gram-negative, motile bacteria isolated from deep subsurface sediments mineralized the nitrogen-containing polyaromatic hydrocarbon quinoline under aerobic conditions and transformed quinoline to soluble intermediates under anaerobic conditions . Many aromatic compounds were also able to serve as the sole source of carbon and energy under aerobic conditions . Rapid aerobic mineralization of quinoline at concentrations as low as 0.002 microgram ml-1 indicates that these organisms possess a high-affinity uptake and utilization system, which may reflect the oligotrophic nature of deep subsurface environments . Both bacteria harbored four plasmids of identical size, ranging from 50 to 440 kilobases.

Br J Rheumatol, 1989 Apr, 28(2), 113 - 7
Mortality of Jamaican patients with systemic lupus erythematosus; Harris EN et al.; A retrospective study of all patients with systemic lupus erythematosus (SLE) who died at the University Hospital of the West Indies over a 14-year period is presented . The major cause of death was infection followed by renal failure . Gram-negative organisms were the major microbiological agents causing infections . Side-effects of therapy were common, in particular bone marrow depression and haemorrhage related to anticoagulants . It appears that controlling severe lupus activity without increasing the risk of lifethreatening complications remains an important goal in the treatment of SLE.

Mayo Clin Proc, 1989 Apr, 64(4), 433 - 45
Intensive-care unit experience in the Mayo liver transplantation program: the first 100 cases; Plevak DJ et al.; The first 100 liver transplantations at the Mayo Clinic were performed in 83 patients, who required a total of 917 patient days in the intensive-care unit (ICU) . The mean duration of stay in the ICU was 5.91 days after liver transplantation and 6.15 days for patients who subsequently required readmission to the ICU . During the immediate postoperative period, hypothermia and hyperglycemia invariably occurred . Later during the initial admission or on readmission to the ICU, there arose the possibility of infections and renal insufficiency . Prompt diagnosis and treatment are necessary for hypertension, hypokalemia, severe metabolic alkalosis, fever, altered mental status, oliguria, and signs of graft failure in liver transplant patients . In our patient series, selective bowel decontamination minimized the occurrence of gram-negative and fungal sepsis, and use of antihypertensive agents and correction of coagulopathies may have decreased the risk of intracranial bleeding in patients with hypertension and clotting defects . Anticipation of potential conditions postoperatively and early implementation of treatment are key factors in the successful ICU management of patients who have undergone liver transplantation.

Surg Clin North Am, 1989 Apr, 69(2), 259 - 71
Liver abscesses; Frey CF et al.; Clinical suspicion of a liver abscess mandates an investigation of the liver for evidence of a liver abscess by radionuclide, ultrasound, or CT scan . Amebic abscesses have a lower mortality rate than pyogenic abscesses . Amebic and pyogenic abscesses can be distinguished on the basis of epidemiologic, clinical, and laboratory studies . The definitive studies for identifying amebic liver abscesses are hemagglutinin or gel diffusion studies . Amebic abscess of the liver may be complicated by extension to the lung, with pulmonary complications . Patients suspected to have amebic abscesses require metronidazole . Emetine or chloroquine may be added if there is no response or if the abscess recurs . Unless there is a failure of the amebic abscess to resolve or secondary infection occurs, there is seldom a need to aspirate or drain these abscesses . Pyogenic abscesses should be treated with broad-spectrum antibiotics to cover gram-negative aerobes and anaerobic organisms . All pyogenic abscesses larger than 1.5 cm in diameter should be aspirated, and the aspirate should be Gram stained and cultured . Percutaneous or surgical drainage should then be performed . Operative intervention is required in those patients with intra-abdominal pyogenic infections that are seeding the liver abscess . The marked reduction in the mortality rate of pyogenic liver abscess witnessed in this decade is multifaceted and attributable in part to earlier diagnosis, permitting definitive treatment in a timely fashion, as well as to improved intensive unit care, antibiotic management, and operative technique.

Singapore Med J, 1989 Apr, 30(2), 141 - 4
A retrospective study of community-acquired pneumonia in Hong Kong with special reference to the choice of antibiotics; Cohen MA et al.; The records of patients admitted to the Prince of Wales Hospital with a presumptive diagnosis of pneumonia over a one-year period were reviewed retrospectively . Forty-four patients fulfilled our diagnostic criteria for community-acquired pneumonia and were included in the survey . The initial choice of antibiotics was reasonably uniform: a penicillin in 75%; erythromycin in 9% and other antibiotics in 16% . Lack of clinical improvement resulted in a change of antibiotics in 15 patients (34%) . Overall 23 patients (52%) responded to a penicillin, 12 patients (27%) responded to erythromycin, and 8 patients (18%) responded to other antibiotics . This suggests that the majority of the causative organisms were sensitive to penicillin, but a significant number of patients required drugs which were effective against 'atypical' and gram negative organisms . One elderly patient, who also suffered from severe chronic airflow obstruction, died after a grand mal seizure which might have been partly related to theophylline toxicity (Mortality 2%) . The investigations performed confirmed or strongly suggested the identity of the causative organism in only 9 patients (21%), and had little or no influence in therapeutic decisions . Although empirical antibiotic treatment was reasonably successful, frequent changes were required, and the lack of firm data on the aetiology indicates that a detailed prospective study is needed.

J Bacteriol, 1989 Apr, 171(4), 2222 - 5
Translation of merD in Tn21; Lee IW et al.; All four sequenced examples of the mercury resistance (mer) operon of gram-negative bacteria have a promoter-distal reading frame, merD, whose removal has little effect on the resistance phenotype and whose translation has not previously been observed . Using merD-lacZ protein fusions, we show that merD is translated . However, Hg(II)-induced merD expression, as measured by beta-galactosidase activity and immunoblotting, is 10- to 15-fold lower than that of fusions to the gene immediately preceding it, merA.

J Bacteriol, 1989 Apr, 171(4), 2028 - 32
Characterization of lipopolysaccharide from Myxococcus xanthus by use of monoclonal antibodies; Fink JM et al.; Lipopolysaccharide is a major constituent of the cell surface of the gram-negative procaryote Myxococcus xanthus . We have purified lipopolysaccharide from M . xanthus and have shown by silver staining that the lipopolysaccharide contains a heterogeneous population of molecules which migrate as a broad low-molecular-mass band (approximately 5 kilodaltons) and as a stepladder of about 30 higher-molecular-mass bands (15- to 70-kilodalton range) . The broad band consists of lipopolysaccharide molecules with just lipid A and core regions . The stepladder bands contain lipopolysaccharide molecules with lipid A, core regions, and various numbers of O-antigen units . Monoclonal antibodies generated against the cell surface of developing M . xanthus cells (J . S . Gill and M . Dworkin, Proc . Natl . Acad . Sci . USA 84:4505-4508, 1987) were used to help characterize the lipopolysaccharide molecules . Five monoclonal antibodies bound to carbohydrate epitopes on the stepladder but not to the broad band, indicating that these monoclonal antibodies recognize carbohydrates on the O antigen of the lipopolysaccharide molecules . Four of these five monoclonal antibodies bound to doublet bands in the stepladder, while the other monoclonal antibody bound to singlet bands in the stepladder . One monoclonal antibody bound to a carbohydrate epitope on both the broad band and the stepladder, indicating that it bound to the core of the lipopolysaccharide.

Am J Vet Res, 1989 Apr, 50(4), 480 - 2
Preferential decay of passively acquired immunoglobulins recognizing shared gram-negative core antigens in neonatal swine; Tyler JW et al.; Serum immunoglobulins of the IgG isotype recognizing common gram-negative cell core epitopes were serially measured by use of a direct ELISA on blood obtained from 10 neonatal swine . An R-mutant Escherichia coli (strain J5) was used as a plate antigen . Total serum IgG was measured by use of radial immunodiffusion . Half-lives of core antigen-specific IgG (6.81 days) and total serum IgG (14.85 days) were dramatically different (P less than 0.01).

J Biol Chem, 1989 Mar 15, 264(8), 4434 - 41
Lipopolysaccharide interaction with lysozyme . Binding of lipopolysaccharide to lysozyme and inhibition of lysozyme enzymatic activity; Ohno N et al.; Experiments have been carried out to characterize the binding of lysozyme (LZM) to bacteriol lipopolysaccharide (LPS) . The formation of LPS.LZM complexes can be readily demonstrated using either physical-chemical separation techniques or a radiolabeled photoaffinity LPS probe . The binding affinity of LZM for LPS has been estimated to be approximately 10(8) liters/mol . Binding of LPS results in loss of LZM enzymatic activity by a noncompetitive inhibition, as assessed by either particulate or soluble substrates . This interaction of LPS with LZM is dictated primarily by hydrophobic interactions and appears to be a general property of both constituents . Binding can be demonstrated with LZM of both human and avian sources, as well as with LPS isolated from a variety of Gram-negative organisms . The addition of LPS to biologically relevant fluids containing LZM results in dose-dependent inhibition of LZM enzymatic activity suggesting that such interactions may have relevance in Gram-negative infections . Finally LZM has been shown to reduce the endotoxic activity of LPS as assessed by gelation of Limulus amoebocyte lysates.

Plasmid, 1989 Mar, 21(2), 99 - 112
Host-specific effects of the korA-korB operon and oriT region on the maintenance of miniplasmid derivatives of broad host-range plasmid RK2; Schmidhauser TJ et al.; Two genetic determinants are sufficient for small derivatives of broad host-range plasmid RK2 to replicate in different Gram-negative bacteria: trfA, which encodes a replication initiator, and oriV, the origin of replication . In this study, nonessential RK2 determinants in the region encoding oriT, the origin of conjugative transfer, and the korA-korB operon, whose products regulate trfA expression, were tested for their effects on the stability of mini-RK2 plasmids in eight different hosts . We found that determinants of both regions can substantially alter plasmid stability, but the effects are not uniform in all hosts . The results also indicate that the effects of the korA-korB operon extend beyond that of the regulation of trfA transcription . This study further illustrates the different requirements for stable plasmid maintenance in diverse bacteria and the ability of wild-type RK2 to adapt to a variety of intracellular environments . The data also provide further evidence for the involvement of different regions of RK2 for stable maintenance in various hosts.

Kosm Biol Aviakosm Med, 1989 Mar-Apr, 23(2), 75 - 9
{Comparative ecologic study of microbial biocenosis of the lettuce rhizosphere under various conditions of cultivation}; Drugova NA et al.; The qualitative and quantitative composition of microorganisms occurring in the rhizosphere of lettuce plants grown in a Svet hot-house on the Balcanin substrate using humates, canned and silvered water was investigated . Microbiological experiments were carried out during a 300-day study within which 5 plant vegetations took place . In the course of a long-term use of the substrate . Gram-negative bacteria became inhibited, streptomycetes abundant and Penicillium and Trichoderma perdominant . The granulometric composition of the substrate and different photoperiods influenced rhizosphere microorganisms without changing substrate microorganisms . It can be concluded that humates and canned water help maintain microbiological activity of the plant rhizosphere which allows a longer-term use of the Balcanin substrate for plant cultivation in the Svet hot-house.

J Clin Microbiol, 1989 Mar, 27(3), 465 - 73
Cellular fatty acid compositions and isoprenoid quinone contents of 23 Legionella species; Lambert MA et al.; The cellular fatty acid compositions and ubiquinone contents of 182 Legionella strains representing 23 species were determined by capillary gas-liquid chromatography and reverse-phase high-performance liquid chromatography, respectively . Except for the type strain of Legionella erythra (ATCC 35303T), all Legionella species contained large (40 to 90%) amounts of branched-chain fatty acids and only trace to small (less than 0.5 to 5%) amounts of ester-linked hydroxy acids . The 23 species were placed in three major fatty acid groups on the basis of differences in the relative amounts of 14-methylpentadecanoic (Ci16:0), hexadecanoic (C16:1), and 12-methyltetradecanoic (Ca15:0) acids . All Legionella species contained ubiquinones with 9 to 14 isoprene units in the side chains and were divided into five different ubiquinone groups . The species were further differentiated into 16 groups on the basis of qualitative and quantitative differences in their fatty acid compositions and ubiquinone contents . Both of these chemical characteristics can be used to distinguish Legionella species from other gram-negative bacteria and rapidly and accurately identify suspected isolates before serologic and other tests are done.

Drug Chem Toxicol, 1989 Mar, 12(1), 49 - 59
Disposition kinetics of cimetidine and ranitidine in endotoxin pretreated rats; Kaka JS et al.; Cimetidine and ranitidine are used in patients with life-threatening gram-negative infections, endotoxemia and acute stress erosions . Disposition kinetics of cimetidine and ranitidine in endotoxin pretreated rats was investigated . The H2-antagonists were administered intravenously 24 h after endotoxin (10 mg/kg) pretreatment . This endotoxin dosage resulted in 50% mortality in rats . Blood samples (0.25 ml) were collected at different timed intervals . No significant differences were observed in plasma clearance, half-life and volume of distribution between endotoxin pretreated and control rats . Cimetidine is eliminated extensively by the renal route in animals and man with metabolism being a minor process . Ranitidine is metabolized to a large extent (70%) in rats, while in humans this represents a minor process . No significant changes in cimetidine and ranitidine disposition parameters in endotoxin pretreated rats were observed . These results suggest that cimetidine and ranitidine may be used in normal dosages in endotoxemia patients since their pharmacokinetic parameters would not be affected under these circumstances.

J Appl Physiol, 1989 Mar, 66(3), 1059 - 64
Effect of inhaled endotoxin on bronchial reactivity in asthmatic and normal subjects; Michel O et al.; Endotoxins are released from the membrane of Gram-negative bacteria present in the environment and in oral and nasal cavities . They are proinflammatory substances that could participate in bronchial obstruction and hyperreactivity in asthmatic patients . This hypothesis was tested by using bronchial challenge tests with inhaled lipopolysaccharides (LPS) from Escherichia coli 026:B6 (22.2-micrograms total dose) followed by a histamine nonspecific challenge test and compared with a placebo procedure, in which the diluent was substituted for the LPS solution . In doing so we showed that LPS induces a slight but significant (P less than 0.01) bronchial obstruction (measured as forced expiratory volume in 1 s) in asthmatics (n = 8) but not in normal subjects (n = 6) . The histamine hyperresponsiveness, expressed as the dosage of histamine necessary to decrease the bronchial specific conductance by 50%, was increased 5 h after LPS inhalation in asthmatics (P less than 0.05) but not in normal subjects . This effect of LPS on bronchial obstruction and hyperresponsiveness was observed in extrinsic (n = 6) as well as in intrinsic (n = 2) asthma.

Arch Dis Child, 1989 Mar, 64(3), 388 - 94
Outcome of neonatal necrotising enterocolitis: results of the BAPM/CDSC surveillance study, 1981-84; Palmer SR et al.; Neonatologists in 100 special care baby units in the United Kingdom and Ireland collaborated in a four year surveillance study of neonatal necrotising enterocolitis . The average overall annual reporting rate of necrotising enterocolitis for infants in England and Wales was 0.3/1000 live births, but ranged from 9.5/1000 live births in infants weighing less than 1000 g at birth to 0.2/1000 live births in infants weighing 2500 g or more . There were more deaths among girls, infants who weighed less than 1500 g at birth, those whose bleeding was abnormal or who had low peripheral platelet counts, infants with Gram negative bacteraemia, and very low birthweight infants who developed it during the first few days of life . In both boys and girls, and in all birthweight groups, operation was associated with increased mortality.

Acta Chir Scand, 1989 Mar, 155(3), 145 - 9
Depression of plasma endotoxin levels during gram-negative septicemia subsequent to moderate trauma; Rokke O et al.; The influence of a moderate, standardized trauma on bacterial and endotoxin kinetics in post-traumatic Escherichia coli septicemia was studied in a porcine model . Septicemia was induced by intravenous infusion of live E . coli (2.5 x 10(9) cfu/kg, rough:K5:H6) into 14 piglets . Seven of these animals had been exposed to moderate trauma 48 hours previously . Following the E . coli infusion, cardiovascular signs of severe septicemia appeared in all the piglets, associated with gradual increase in the blood bacterial count and the plasma levels of endotoxin . This increase was significantly less pronounced in the animals with prior moderate trauma than in the nontraumatized animals . Thus no depression of the host defense system was demonstrable 2 days after moderate trauma.

Mod Pathol, 1989 Mar, 2(2), 85 - 9
Factors contributing to mortality in lung transplant recipients: an autopsy study; Cagle PT et al.; Despite improved surgical techniques and advances in immunosuppressive therapy, posttransplant mortality rates remain significantly high in lung transplant patients . Since 1985, 3 of 6 single lung recipients, 3 of 3 double lung recipients, and 4 of 7 heart-lung recipients have died and undergone autopsy . We reviewed the autopsy findings in these patients to determine the type and frequency of pathologic processes associated with mortality . One or more infectious processes was found in every patient at autopsy . Gram-negative bacterial pneumonia and sepsis, found in 7 and 8 of 10 autopsy cases, respectively, were by far the most frequent contributing factors to mortality . Epstein-Barr virus infection was demonstrated in one patient using polymerase chain reaction amplification . Acute transplant rejection was found in only 2 patients and therefore is a much less common factor in the death of lung transplant recipients . Diffuse alveolar damage occurred in 6 patients and bronchiolitis obliterans occurred in 3 patients . These latter two processes may have different etiologies in different patients.

Vet Microbiol, 1989 Mar, 19(3), 283 - 9
Antibodies to Escherichia coli J5 core glycolipids in gnotobiotic and conventionally reared piglets; McVey DS et al.; Antibody titers to Escherichia coli J5 endotoxin core glycolipids were determined in 5-week-old, unvaccinated, conventionally reared and gnotobiotic pigs in a micro-ELISA . Both groups of pigs had naturally occurring antibody of all major isotypes . The conventionally reared pigs had significantly (P less than 0.05) more serum antibody to J5 endotoxin glycolipids (geometric mean titer, GMT = 2.07) than gnotobiotes (GMT = 0.85) . This relationship was not necessarily true in fluids from various anatomical structures . In gnotobiotes, mucosal-associated, anti-J5 glycolipid immunoglobulin was primarily IgM . In conventionally reared piglets, the mucosal-associated, anti-J5 glycolipid antibody contained more IgA . Such naturally occurring antibody could confer partial to complete immunity from challenge with Gram-negative bacteria.

Pediatr Infect Dis J, 1989 Mar, 8(3), 155 - 9
Cerebrospinal fluid interleukin 1-beta and tumor necrosis factor concentrations and outcome from neonatal gram-negative enteric bacillary meningitis; McCracken GH Jr et al.; Concentrations of interleukin 1-beta and tumor necrosis factor in cerebrospinal fluid (CSF) of 42 neonates with Gram-negative enteric bacillary meningitis were determined and correlated with outcome and other CSF indices . At the time of diagnosis interleukin 1-beta was detected in CSF of 40 infants and peak concentrations correlated significantly with outcome from disease . Days of interleukin 1-beta concentrations of greater than 200 pg/ml or greater than or equal to 20 pg/ml were significantly correlated with days that CSF cultures were positive and that K1 antigen and endotoxin were detected in CSF . Tumor necrosis factor activity was detected in CSF of 25 of 27 infants; there was no correlation between outcome and CSF tumor necrosis factor concentrations . The results provide a possible rationale for therapeutic intervention to improve outcome.

J Exp Med, 1989 Mar 1, 169(3), 823 - 32
Endotoxin and tumor necrosis factor challenges in dogs simulate the cardiovascular profile of human septic shock; Natanson C et al.; Survivors of both human and animal bacterial shock develop a characteristic pattern of progressive changes in cardiovascular function over a period of 7-10 d . In this present study, we examined whether endotoxin (a product of Gram-negative bacteria) or TNF (a cytokine released from macrophages) could reproduce the same complex cardiovascular changes observed in septic shock over a period of 7-10 d . To test this hypothesis, we implanted a thrombin-fibrin clot containing purified endotoxin from E . coli into the peritoneal cavity of eight dogs, and infused TNF into eight different dogs . Over the next 10 d, serial simultaneous heart scans and thermodilution cardiac outputs were performed in these awake nonsedated animals . By day 2 after challenge with either endotoxin or TNF, animals developed a decrease (p less than 0.05) in both mean arterial pressure and left ventricular ejection fraction . With fluid resuscitation, animals manifested left ventricular dilatation (increased {p less than 0.05} end diastolic volume index), increased or normal cardiac index, and decreased or normal systemic vascular resistance index . In surviving animals, these changes returned to normal with 7-10 d . The time course of these changes was concordant (p less than 0.05) with that previously described in a canine model of septic shock using viable bacteria . During the 10-d study, control animals receiving sterile clots or heat-inactivated TNF had not significant changes in hemodynamics . The results from this canine model demonstrate that either endotoxin or TNF alone can produce many of the same hemodynamic abnormalities seen in human septic shock and in a canine septic shock model induced by live bacteria . These findings support the hypothesis that the action of endogenous mediators (TNF) responding to bacterial products (endotoxin) is the common pathway that produces the serial cardiovascular changes found in septic shock.

Oral Microbiol Immunol, 1989 Mar, 4(1), 52 - 6
Characterization of selected gram-negative oral microorganisms by SDS-PAGE; Kinder SA et al.; SDS-PAGE analysis of bacterial cell proteins was used to examine a diverse group of oral microorganisms . Most of the species examined, including physiologically similar microorganisms, produced distinct protein patterns which are characterized by the distribution of their major protein bands . The SDS-PAGE system described provides a rapid, simple and standardized identification technique to be used in consort with routine taxonomic procedures.

Bull Chest Dis Res Inst Kyoto Univ, 1989 Mar, 22(1-2), 11 - 8
{A case report of cat scratch disease which mimicked a metastasis of lung cancer}; Huang CL et al.; A 67-year-old woman to our hospital because of the swelling of lymph nodes in the right inguinal region . Eighteen months before, right lower lobectomy was performed for primary lung cancer (papillary adenocarcinoma) . In September 1986, she noticed the swelling of lymph no does in the right inguinal region and low grade fever . At first she was suspected to have a metastasis of the lung cancer . Lymph node biopsy was performed . The pathological examination revealed purulent lymphadenitis with granulomatous change, and Gram-negative Warthin Starry stain positive rods were found . In addition, she admitted a history of keeping cats fourteen years . Therefore the diagnosis of cat scratch disease was made . Six months later, she became free of the associated symptoms . We feel pathological confirmation of suspicious lymph nodes is important in the treatment of patients with history of malignancy.

J Clin Invest, 1989 Mar, 83(3), 970 - 7
Bacterial lipopolysaccharides prime human neutrophils for enhanced production of leukotriene B4; Doerfler ME et al.; Neutrophils can be "primed" for an enhanced respiratory burst by lipopolysaccharide (LPS) in concentrations measurable in patients with septic shock . Leukotriene B4 (LTB4) is the primary eicosanoid product of neutrophils and is felt to be a mediator of host defense and inflammation . We investigated the in vitro effects of LPS on neutrophil production of LTB4 and the omega-oxidation metabolites of LTB4 . Incubation of neutrophils with LPS in concentrations ranging from 0.01 to 100 ng/ml did not result in production of LTB4 or metabolites in the absence of a second stimulus . Priming neutrophils with LPS and then stimulating with opsonized zymosan, phorbol-myristate-acetate or a low concentration of the calcium ionophore A23187 resulted in enhanced production of LTB4 . LPS priming of neutrophils occurred in a concentration dependent manner . LPS did not result in LTB4 production in response to the chemoattractant peptide FMLP . LPS priming of neutrophils had no effect on cytosolic calcium concentrations of resting or zymosan-stimulated cells . These results suggest that LPS might effect host defense and tissue injury by potentiating the effect of other stimulants on neutrophil production of LTB4 . This LPS induced enhancement may represent an important pathogenetic pathway in patients with gram negative sepsis.

Microb Pathog, 1989 Mar, 6(3), 159 - 74
Isolation of the outer membrane of Branhamella catarrhalis; Murphy TF et al.; The emergence of Branhamella catarrhalis as an important human pathogen has stimulated interest in investigations of the outer membrane (OM) of the bacterium . In this study, the OM of B . catarrhalis was isolated and partially characterized . Radiolabelled cells were lysed and fractionated by isopycnic centrifugation in a continuous sucrose gradient . Five fractions were identified . Fraction A consisted of OM fragments of varying density . Fractions B and C were OM of a discrete density containing some cytoplasmic membrane . Fraction D was cytoplasmic membrane and Fraction E contained smaller less dense fragments of cytoplasmic membrane . The protein composition of the Branhamella OM is typical for that of Gram-negative bacteria in that approximately 10 to 20 proteins were present with six to eight of these proteins predominating . Having isolated and partially characterized the OM by sucrose density centrifugation, five simpler techniques for isolating OM were employed and the preparations compared to OM isolated on the gradient . Techniques that are based on differential detergent solubility of OM and cytoplasmic membrane were ineffective in isolating OM of B . catarrhalis . By contrast, techniques that involved collection of OM vesicles were successful in isolating OM of B . catarrhalis . Collection of vesicles from broth culture supernatants and EDTA-heat-induced vesicles were identified as convenient and reliable methods for isolating OM . Isolating and partially characterizing the OM of B . catarrhalis represents an initial step in a systematic study of outer membrane antigens of the bacterium.

J Surg Res, 1989 Mar, 46(3), 195 - 9
Complement levels in septic primates treated with anti-C5a antibodies; Hangen DH et al.; During gram-negative sepsis it is known that endotoxin activates complement by the alternate pathway . The complement anaphylatoxin C5a, a result of this activation, is thought to play a key role in attracting and activating neutrophils in the lungs, leading to the adult respiratory distress syndrome . Complement levels were measured in primates made septic by Escherichia coli infusions . Anti-human C5a antibodies were administered to study their effect on neutrophil-mediated lung injury . Control (I), septic (II) and septic + anti-C5a antibody (III) groups (n = 4) were studied . The antibody-treated group (III) demonstrated a significant attenuation of septic shock and pulmonary edema as has been previously reported . All complement profiles were corrected for varying hemoglobin concentrations . C3, C4, and C5 levels were measured by radial immunodiffusion and were depleted in both septic groups . Once the levels were depleted from the plasma, they did not recover . The depletion of C4 indicates that classical pathway activation also occurred . C3a, C4a, and C5a levels were measured by radioimmunoassay . Significantly increased peak levels were reached in the septic groups 15 min after initiation of the E . coli infusion . There were no significant differences in early peak C3a and C4a levels between groups II and III . However, the mean peak C5a level in group III (anti-C5a antibodies) was 42% lower than that in group II, and after this early peak, C5a levels were not elevated above control levels in group III . The antibody to human C5a was thus shown to be cross-reactive with primate C5a and was specific since C3a and C4a levels were not decreased in group III.(ABSTRACT TRUNCATED AT 250 WORDS)

Arch Surg, 1989 Mar, 124(3), 339 - 41
Comparison of antibody response with delayed hypersensitivity in severely injured patients; Hershman MJ et al.; Twenty-seven severely injured patients had antibody response to gram-negative organisms measured of whom 25 also had skin testing . Twenty-three patients (92%) were anergic at admission . Injury Severity Scores were greater in patients who remained anergic for three weeks compared with patients whose skin tests became positive . Patients with major infection had longer anergy duration than uninfected patients . Twenty-two (81%) of 27 anergic patients mounted antibody responses . Initial bacterial contamination determined the clinical outcome and antibody response . Six of eight patients with moderate contamination mounted IgM responses to all organisms and/or an IgG response to Escherichia coli, and they remained uninfected . Of the other two patients, one died, and the other developed chronic infection . Seven of eight patients with heavy contamination developed major sepsis despite mounting several antibody responses . Six patients without contamination mounted no antibody responses and remained uninfected.

FEMS Microbiol Lett, 1989 Mar, 49(1), 59 - 63
Molecular cloning of a gene encoding a Chlamydia psittaci 57-kDa protein that shares antigenic determinants with ca . 60-kDa proteins present in many gram-negative bacteria; Menozzi FD et al.; In order to develop reagents to study the immune response of guinea pigs to infection by Chlamydia psittaci guinea pig inclusion conjunctivitis strain (GPIC), we constructed a plasmid clone bank with C . psittaci DNA . One of the recombinant clones isolated produced large amounts of a 57-kilodalton (kDa) protein that was immunoreactive with sera from GPIC infected guinea pigs . While investigating this recombinant protein, we discovered that all the Gram-negative bacteria analyzed so far have immunoreactive proteins of similar size . This protein seems to be a 'common antigen' already described in various Gram-negative bacteria.

J Immunol Methods, 1989 Feb 24, 117(2), 175 - 80
Quantitative assay of a human monoclonal IgM antibody (HA-1A) in human serum; Khazaeli MB et al.; A rapid, specific and sensitive radiometric assay was developed capable of quantitating serum levels of HA-1A, a human IgM monoclonal antibody to endotoxin . 'Private' anti-idiotypic murine monoclonal antibodies were produced and utilized in the assay to avoid cross-reactivity with normal human IgG, IgM, IgA, IgE or IgD . The presence of E . coli or gram-negative lipopolysaccharide in the sera did not affect the ability of the assay to detect HA-1A . The sensitivity of the assay was calculated to be 25 ng/ml with an interassay coefficient of variation of less than 10% . In one patient given 100 mg of HA-1A, peak serum concentration was 101.5% of the predicted value with a mean plasma half life of 24.5 h . This assay will be useful in establishing the pharmacokinetics of HA-1A and in monitoring serum levels during phase II and phase III clinical trials.

Surgery, 1989 Feb, 105(2 Pt 1), 200 - 6
Changes in the levels of endogenous antioxidants in the liver of mice with experimental endotoxemia and the protective effects of the antioxidants; Sugino K et al.; Gram-negative bacterial endotoxin (lipopolysaccharide) induces hepatic damage in mice caused by lipid peroxidation, and administration of antioxidants (coenzyme Q10 and alpha-tocopherol) suppresses this lipid peroxidation, preserves energy metabolism, and enhances the survival of endotoxin-administered mice . Therefore experiments were done to determine whether experimental endotoxemia in mice affected the levels of endogenous antioxidants and whether treatment with antioxidants altered these levels . Endotoxin produced decreases in hepatic endogenous antioxidants with time: reduced coenzyme Q9, alpha-tocopherol, and reduced glutathione were lowered to 29.9%, 27.1%, and 45.4% of the control levels, respectively, 24 hours after administration . Exogenously administered coenzyme Q10 (oxidized) accumulated in the liver and showed a maximal plateau between 8 and 16 hours after injection when 82% of coenzyme Q10 was converted to the reduced form . Coenzyme Q10 administered with endotoxin totally or partially prevented the decreases in these endogenous antioxidants, and furthermore, total coenzyme Q10 and the reduced form, both at levels of approximately 30 mg protein, were consumed during the period of elevated lipid peroxidation, 16 hours after endotoxin injection . These results indicate that coenzyme Q10 acts in vivo as an antioxidant after it has been converted to the reduced form . alpha-Tocopherol administered also showed an 84-fold accumulation in the liver 8 hours after injection, completely preventing any decrease in endogenous reduced coenzyme Q9 and partially preventing reduction of glutathione, which indicated an in vivo antioxidant action of alpha-tocopherol . These results support the assumption that administered coenzyme Q10 or alpha-tocopherol functions cooperatively with endogenous antioxidants to prevent tissue damage caused by lipid peroxidation in endotoxemia.

J Clin Microbiol, 1989 Feb, 27(2), 231 - 5
Capnocytophaga canimorsus sp . nov . (formerly CDC group DF-2), a cause of septicemia following dog bite, and C . cynodegmi sp . nov., a cause of localized wound infection following dog bite; Brenner DJ et al.; CDC group DF-2 is the vernacular name given to a slow-growing gram-negative bacterium that causes septicemia and meningitis in humans . Infections frequently (one-third of cases) occur following dog bites or close contact with dogs or occasionally with cats . Splenectomy and alcoholism appear to be strong predisposing factors for DF-2 infection . In addition to 150 DF-2 strains received for identification, we received 9 DF-2-like strains; 6 were isolated from wound or eye infections, 3 of which were associated with dog bites and 1 of which was associated with a cat scratch, and 3 were isolated from dog mouths . The major characteristics of DF-2 include production of acid but no gas from lactose and maltose and usually D-glucose; positive reactions for oxidase, catalase, arginine dihydrolase, gliding motility, and o-nitrophenyl-beta-D-galactopyranoside; growth enhanced by serum and by incubation in a candle jar atmosphere; and negative reactions for sucrose, raffinose, inulin, melibiose, nitrate reduction, indole, and growth on MacConkey agar . DF-2-like strains had the same characteristics, except that acid was formed from sucrose, raffinose, inulin, and melibiose . By the hydroxyapatite method, DNAs from 12 DF-2 strains were 88% related in 60 degrees C reactions and 84% related in 75 degrees C reactions . Related sequences contained 0.5 to 1.5% unpaired bases (divergence) . Three DF-2-like strains were 73 to 80% related at 60 degrees C (with 2.0 to 2.5% divergence) and 68 to 75% related at 75 degrees C . The relatedness of DF-2 and DF-2-like strains was 19 to 31% at 60 degrees Celsius and 13 to 19% at 75 degrees Celsius . The relatedness of DF-2 and DF-2-like strains to Capnocytophaga species was 4 to 7% . The DNA relatedness date indicate that eh DF-2 and the DF-2-like strains are separate, previously undescribed species . Both groups are phenotypically and genetically distinct from Capnocytophaga species, although they do share several characteristics with Capnocytophaga species, including cellular morphology, gliding motility, cellular fatty acid composition, enhancement of growth in a candle jar atmosphere, and G+C content . The new species differ from Capnocytophaga species by their positive oxidase and catalase reactions . We chose to avoid creating a new genus and proposed the names Capnocytophaga canimorsus sp . nov . for group DF-2 and C . cynodegmi sp . nov . for the DF-2-like strains.

J Bacteriol, 1989 Feb, 171(2), 952 - 7
Increase of ornithine amino lipid content in a sulfonolipid-deficient mutant of Cytophaga johnsonae; Pitta TP et al.; The gram-negative gliding bacterium Cytophaga johnsonae contains not only large quantities of unusual sulfonolipids but also, as we report here, a second class of unusual lipids . These lipids were detected and quantified by two-dimensional thin-layer chromatography of lipids from cells grown in the presence of {14C}acetate and shown by chemical studies to be alpha-N-(3-fatty acyloxy fatty acyl)ornithines . Like the sulfonolipids, these ornithine lipids were localized in the outer membrane (whereas phosphatidylethanolamine was the predominant lipid of the inner membrane) . In a sulfonolipid-deficient mutant, the missing lipid was replaced, specifically, by an increased amount of ornithine lipid . Cells grown in liquid media contained predominantly ornithine lipids with nonhydroxylated residues in the O-fatty acyl position . In contrast, surface-grown cells contained a high proportion of ornithine lipids in which the O-fatty acyl group was 3-hydroxylated . The sulfonolipids and ornithine lipids are apparently coregulated in the sense that, regardless of perturbations caused by mutation or growth conditions, their total amounts remain constant at 40% of total cell lipid.

Laryngorhinootologie, 1989 Feb, 68(2), 81 - 7
{Lyme borreliosis and its significance for the ENT physician}; Diehl GE et al.; Lyme disease is a tick-borne multisystemic Borrelia infection to which the following diseases belong: erythema migrans, lymphadenosis benigna cutis, lymphocytic meningoradiculitis (Bannwarth's syndrome), Lyme-arthritis and acrodermitis chronica atrophicans . The infection rate of ticks with Borrelia Burgdorferi in Germany amounts to 13.6% compared to the infection with the European spring summer meningoencephalitis virus with 1.1% . Recent investigations show that lipopolysaccharides and interleukin-1 play an important role in the pathogenesis of Lyme disease . Lipopolysaccharides (LPS) are a constitutive part of the outer wall of gram negative bacteria . Its biological activities include pyrogenicity, mitogenicity for lymphocytes and the induction of interleukin-1 (IL-1) . IL-1 is the major macrophage-derived immunoregulatory protein . Lyme disease is characterized by a variety of symptoms which could be explained by the effects of IL-1 on host systems . These symptoms include: fever, malaise, erythema migrans and arthritis . The clinical course can be divided into three stages . Erythema migrans, lymphadenosis benigna cutis and general symptoms characterize the first stage . In the second stage disorders of the heart and the neurological system may follow including Bannwarth's syndrome . 60% of the patients develop facial palsy and 30% of these patients bilateral palsy . In 40% of all cases the facial palsy is the only motor disorder . Other cranial nerves can also be affected . The third stage consists of the Lyme-arthritis, acrodermitis chronica atrophicans and encephalomyelitis . The determination of specific spirochetal antibodies in serum and cerebrospinal fluid (CSF) is the most valuable diagnostic aid for this borreliosis . The CSF examination may also be helpful.(ABSTRACT TRUNCATED AT 250 WORDS)

Nippon Shokakibyo Gakkai Zasshi, 1989 Feb, 86(2), 246 - 52
{Treatment of endotoxin shock due to gram-negative bacteremia using extracorporeal circulation}; Asanuma Y et al.; The effectiveness and significance of the various therapeutic methods of extracorporeal circulation were assessed in an experimental model of septic endotoxin shock . After sepsis was produced in 30 puppies by intravenously injecting 5 X 10(9) CFU/ml/kg of E . coli, on-line plasma exchange, conventional plasma exchange, charcoal plasma perfusion, hemofiltration, and sham perfusion were performed for 2 hr . As a result, 4 of the 5 puppies in the on-line plasma exchange group and one of the 5 in the conventional plasma exchange group survived; the others all died . In the on-line plasma exchange group, the number of E . coli decreased to 2.5 X 10 CFU/ml at the end of the treatment, while it was 1.3 X 10(3) CFU/ml in the sham perfusion group . The endotoxin concentration was also reduced significantly, while plasma opsonin activity was remarkably elevated . These results suggest the significance of achieving the elimination of endotoxin in blood and an increase in plasma opsonin activity simultaneously in the treatment of septic endotoxin shock.

Anaesth Intensive Care, 1989 Feb, 17(1), 49 - 55
Endotoxins and anti-endotoxins (their relevance to the anaesthetist and the intensive care specialist); Brock-Utne JG et al.; Endotoxins (lipopolysaccharides, LPS) are potent bacterial poisons always present within the intestines in considerable amounts . Several pathophysiological conditions such as hypovolaemia, hypoxia, intestinal ischaemia, burns and radiation lead to a breakdown in the barrier and depending upon the extent of the injury, endotoxins enter the systemic circulation in increasing amounts . Antibiotics do not inactivate the endotoxins which continue to exert their toxic effects leading to nausea, vomiting, diarrhoea, fever, disseminated intravascular coagulation, vascular collapse and organ failure . When nonabsorbable antibiotics are given prior to the insult, systemic endotoxaemia is prevented . Immunotherapy, using anti-lipopolysaccharide IgG, inactivates plasma endotoxins, destroys gram-negative bacteria and opsonises them and may become a major form of therapy . An outline of endotoxin and anti-lipopolysaccharide and its importance to the anaesthetist and intensive care specialist is presented.

Infect Immun, 1989 Feb, 57(2), 656 - 9
Structural analysis of lipopolysaccharides from Eikenella corrodens by use of murine monoclonal antibodies; Kato T et al.; The structure of lipopolysaccharides (LPSs) of Eikenella corrodens was analyzed with prepared murine monoclonal antibodies . A common core epitope was found in three of seven LPS preparations from E . corrodens strains and Fusobacterium nucleatum ATCC 25586 . Four E . corrodens LPSs were found to possess an O-side-chain epitope which cross-reacted with LPSs from Fusobacterium necrophorum ATCC 25286 and Capnocytophaga ochracea M-12 . Lipid A of E . corrodens LPS shared an epitope common among LPSs from various gram-negative rods.

Mol Gen Genet, 1989 Feb, 215(3), 416 - 24
Genetic structure, function and regulation of the transposable element IS21; Reimmann C et al.; The IncP plasmid R68.45 and other plasmids carrying tandem repeats of the insertion sequence IS21 {= (IS21)2} produce replicon fusions via transposition at high frequencies in Escherichia coli and other gram-negative bacteria, whereas plasmids with a single IS21 copy, e.g . R68, give replicon fusions rarely . The 2131 bp nucleotide sequence of IS21 was determined; at the ends there were 11 bp inverted repeats with one mismatch . Two adjacent open reading frames, istA and istB, were located on one DNA strand of IS21 . In E . coli maxicells, polypeptides of 46 kDa (the istA gene product) and 30 kDa (the istB gene product) were expressed by (IS21)2 plasmids, but not by IS21 plasmids . Genetic analysis of (IS21)2 plasmids indicates that the IS21-IS21 junctions form a promoter, which initiates transcription of the istAB operon in one of the two IS21 elements . A single IS21 element fused to an inducible external tac promoter expressed both proteins after induction, but did not promote effective replicon fusion, unless an IS21-IS21 junction (the preferred site for IS21 transposase action) was also present on the plasmid carrying the tac-IS21 construct . The sequences located between the IS21 elements in (IS21)2, 3 bp in R68.45 or 2 bp in pME28, were not recovered in the replicon fusion products . Homologous recombination between the directly oriented IS21 elements in the fusion products led to plasmids with a single IS21 insertion . Analysis of the latter showed that IS21 had a low, but not totally random specificity of insertion and created target duplications of 4 bp (occasionally 5 bp).(ABSTRACT TRUNCATED AT 250 WORDS)

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1989 Feb, 187(3), 181 - 208
{Colony count and LPS content of gram-negative bacteria in cold storage foods and water}; Weber-Frick C et al.; In storage trials, the multiplication of gram negative bacteria was monitored by means of both colony counts and LPS-formation as determined by the three LAL tests methods (the Capillary test, the "Mini" Endotoxin test and the Coatest endotoxin method) . The detection limits of the colony forming units which could be determined by the three LAL tests were initially set in model experiments in which beef (M . cleidooccipitalis) was variously inoculated with stationary-phase cells of Pseudomonas sp . previously grown at 30 degrees C . In all three methods, measurable amounts of LPS were possible only at colony counts above 10(3)/ml, g or cm2 . The detection limits for colony counts of vacuum packed, aerobically stored beef muscle (Caput long . of M . triceps brachii) were found to lie between 1.0 x 10(3) and 1.0 x 10(4)/g for the Capillary test and the Coatest endotoxin method, and 5.0 x 10(4)/g for the "Mini" Endotoxin test . In the case of poultry carcasses the detection limits lay between 2.0 x 10(1) and 7.0 x 10(2)/cm2 and were thus considerably lower than for beef . With very low levels of bacterial loads, substrate interference in the LPS-LAL reaction must be taken into account . Linear regression analysis gave satisfactory correlation between the concentration of LPS and colony forming units for beef, poultry carcasses, ground beef and mixed salad . An acceptable proportionality was established only for beef and poultry carcasses . Potable water, stagnant for a fortnight in an experimental piping system and sampled at five different points, showed significant regrowth of oligocarbotolerant aquatic bacteria . However, only very low levels of LPS could be determined . All three LAL test methods can be recommended for rapid determination of the load of gram negative bacteria in meat and meat products.

Vaccine, 1989 Feb, 7(1), 29 - 33
Novel low-molecular-weight synthetic vaccine against foot-and-mouth disease containing a potent B-cell and macrophage activator; Wiesmuller KH et al.; Most synthetic peptide vaccines described to date are effective only in combination with proteins and Freund's adjuvant . The work describes a novel completely synthetic virus peptide vaccine, which consists of a synthetic activator of B cells and macrophages, covalently linked to an amphiphilic alpha-helical T-cell epitope . The low-molecular-weight vaccine of 3.4 kDa developed against foot-and-mouth disease virus (FMDV) is composed of a synthetic VP1 (135-154) with a sequence homologous to an FMDV protein and the adjuvant tripalmitoyl-S-glyceryl-cysteinylserylserine (P3CSS) . P3CSS is the synthetic analogue of the N-terminal part of the lipoprotein from Gram-negative bacteria . The antigenic determinant VP1 (135-154) is an alpha-helix as shown by circular dichroism . The resulting novel type of vaccine tripalmitoyl-S-glyceryl-cysteinylserylseryl-FMDV-VP1 (135-154) induces a long-lasting high protection against foot-and-mouth disease and serotype-specific virus-neutralizing antibodies in guinea-pigs after a single administration without any additional adjuvant or carrier . In contrast to other simple fatty acid conjugates this new type of vaccine contains a built-in adjuvant with high affinity to both B and T lymphocytes.

Vnitr Lek, 1989 Jan, 35(1), 9 - 15
{Clinical evaluation of granulocyte concentrates in the treatment of malignant disorders of hematopoiesis}; Mocikova K et al.; The authors evaluate in a retrospective, non-randomized two-year study the action of 57 granulocyte concentrates from a Fenwal CS 3000 separator in 12 patients used for induction therapy of acute leukaemia, malignant lymphoma and agranulocytosis, as compared with a control group of 18 patients without concentrates . The indication for selection was granulocytopenia of less than 0.5.10(9)/l, temperatures resistant to antibiotics for more than 48 hours, mainly gram-negative sepsis and a severe localized infection . A significant reduction of febrile days was achieved, to 6.08 as compared with 12.44 (p less than 0.001) along with cure of the infection . Survival, evaluated on the 21st day of the investigation in the treated group 66.6%, as compared with 61.1%, and the percentage of complete remissions 58% as compared with 55.56%, were not statistically significant (p greater than 0.05) . The mean one-hour rise of granulocytes by 0.37 x 10(9)/l in the recipients had no clinical impact . With the exception of one patient a relationship was observed between the favourable action of transfusions and the trend of recovery of the patient's granulopoiesis and the onset of remission of the disease . Minor pyretic and allergic reactions occurred in three patients (5.2%).

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