Am J Vet Res, 1989 Jan, 50(1), 126 - 30 Humoral recognition of lipopolysaccharide core antigens of gram-negative bacteria in neonatal swine; Tyler JW et al.; Serologic recognition of common lipopolysaccharide core antigens has been related to enhanced resistance to gram-negative bacterial disease in several species . Class-specific titers (IgG, IgM) were determined by direct ELISA, using intact Escherichia coli (J5) as a plate antigen . Serum samples were obtained from 224 neonatal swine between the ages of 36 and 60 hours . The mean (+/- SEM) log10 IgG titer against gram-negative core antigens was 1:1,713 +/- 0.4718 and the mean log10 IgM titer was 1:202 +/- 0.5644 . The IgG titer was directly related with litter size, birth weight, and serum total IgG concentration; IgM titer was directly related with dam parity and serum total IgG concentration. J Bacteriol, 1989 Jan, 171(1), 321 - 8 Isolation and nucleotide sequencing of an aminocyclitol acetyltransferase gene from Streptomyces rimosus forma paromomycinus; Lopez-Cabrera M et al.; A gene (aacC7) encoding an aminocyclitol 3-N-acetyltransferase type VII {AAC(3)-VII} from Streptomyces rimosus forma paramomycinus NRRL 2455 was cloned in the Streptomyces plasmid pIJ702 and expressed in Streptomyces lividans 1326 . Subcloning experiments located the aacC7 structural gene on a 1.05-kilobase DNA sequence . The direction of transcription of aacC7 was determined by using riboprobes synthesized in vitro from a DNA fragment internal to the gene . A DNA segment encoding the AAC(3)-VII activity and comprising 1,495 base pairs was sequenced . The aacC7 gene was located in an open reading frame of 864 base pairs that encoded a polypeptide of Mr 31,070, consistent with the Mr (32,000) of the AAC(3)-VII enzyme as determined by physicochemical methods . High-resolution S1 nuclease mapping suggested that transcription starts at or near the A residue of the ATG initiator codon . A DNA fragment from the 5' region of aacC7 had promoter activity in the promoter-probe plasmid pIJ486 . The -10 and -35 regions of this fragment showed limited sequence resemblance to other Streptomyces promoters . The primary structure of the AAC(3)-VII enzyme showed strong homology with those of the AAC(3)-III and AAC(3)-IV enzymes encoded by plasmids in gram-negative bacterial genera . Upstream of the aacC7 gene was an open reading frame of 357 nucleotides which did not appear to be involved in controlling the expression of the aacC7 gene. J Steroid Biochem, 1989 Jan, 32(1A), 27 - 34 Isolation and characterization of a 50 kDa testosterone-binding protein from Pseudomonas testosteroni; Thomas JE et al.; A testosterone-binding protein (Mr = 50,500) has been isolated from the Gram-negative bacterium Pseudomonas testosteroni . The protein was partially purified by a combination of ion exchange chromatography and chromatofocusing . Final purification was achieved by electroelution of the 50 kDa protein from SDS-polyacrylamide gels . Following renaturation from a diluted solution of guanidine-HCl, specific binding of {3H}testosterone to the purified protein was observed . The native protein has a pI of 6.8 . It appears to contain 428 amino acids, 39% of which are hydrophobic . There is only one cysteine residue . Both chymotrypsin and V8 protease were used to produce peptide maps of the protein for use in future identification . The first 10 amino acids situated at the N-terminal of the protein were Ser-Pro-Phe-Asp-Leu-Arg-Pro-Leu-Ser-Gly . Testosterone binding to the protein was saturable at approximately 3.8 nmol/mg protein; the binding constant was approximately 25 nM . Unlabelled testosterone, androstenedione, 5 alpha-dihydrotestosterone and 5 beta-dihydrotestosterone were able to compete for {3H}testosterone bound to the protein; 17 beta-estradiol also competed for {3H}testosterone but to a lesser degree . Neither progesterone nor desoxycorticosterone competed for the testosterone-binding site . Binding of testosterone to the protein was stable at pH's ranging from 5.5 to 9.0 and at various temperatures ranging from 4 to 30 degrees C . The protein was unable to metabolize testosterone in either the presence or absence of the cofactor NAD. J Trauma, 1989 Jan, 29(1), 109 - 12 Secondary infection of post-traumatic pulmonary cavitary lesions in adolescents and young adults: role of computed tomography and operative debridement and drainage; Carroll K et al.; Secondary infection of post-traumatic cavitary lung lesions is unusual . This report describes the clinical course of four patients who sustained severe blunt chest trauma and developed pulmonary pseudocysts that became foci for systemic sepsis . All four patients were adolescents or young adults . Hemophilus species and aerobic Gram-negative rods were the predominant pathogens recovered . Computed tomography of the chest was instrumental in establishing the diagnosis in each case . Despite appropriate antibiotic therapy, all four patients remained septic for weeks . One of the patients died as a result of this infectious process . One patient underwent successful operative debridement and drainage of the involved lung and pleural space . Because infected traumatic pseudocysts may not respond like typical lung abscesses to appropriate antibiotic management, early exploratory thoracotomy should be considered in those patients with prolonged fever and pulmonary deterioration. Chest, 1989 Jan, 95(1), 155 - 61 Patterns and routes of tracheobronchial colonization in mechanically ventilated patients . The role of nutritional status in colonization of the lower airway by Pseudomonas species; Niederman MS et al.; Tracheobronchial colonization by Gram-negative bacteria is common in mechanically ventilated patients . Pseudomonas sp are commonly isolated from the lower airways . We hypothesized that Pseudomonas sp would preferentially colonize the lower airway and would be more common in patients with poor nutritional status . We serially collected 75 pairs of upper and lower respiratory tract cultures from 14 patients treated with mechanical ventilation for at least one week, examined patterns of airway colonization and routes of bacterial entry for Pseudomonas sp and other enteric Gram-negative bacteria (EGNB), and related these findings to host-associated factors, including nutritional status . Pseudomonas sp were the most common species isolates taken from the lower airway, found in nine of 14 patients and in 41.3 percent of all cultures . In contrast to other EGNB, Pseudomonas sp were found significantly (p less than or equal to 0.05) more often in the tracheobronchial tree (31 of 75 cultures) than in the oropharynx (18 of 75 cultures) . Primary colonization of the lower airway by Pseudomonas sp was found in four patients, while other EGNB never followed this pattern when subjects were studied with cultures taken every third day . A host-related factor related to lower airway colonization by Pseudomonas species was poor nutritional status, assessed by a multifactorial index (p less than or equal to 0.01) . We conclude that in mechanically ventilated patients, Pseudomonas sp colonize the lower airway in a different pattern and by a different route from those of other EGNB . The findings that Pseudomonas sp preferentially colonize the tracheobronchial tree may be important for the design of strategies to prevent airway colonization . The recognition that poor nutritional status, a potentially modifiable host-related factor, favors lower airway growth of Pseudomonas sp suggests one direction for future infection-control efforts. Pneumonol Pol, 1989 Jan, 57(1), 29 - 37 {Bacterial infections in pulmonary aspergilloma}; Halweg H et al.; 28 cavernoscopies were carried out in 26 patients . Complete or partial removal of mycetomas was achieved in 18 patients with 20 caverns . Under aerobic conditions Aspergillus fumigatus was cultured from the 28 removed mycetomas in 22 cases (78.6%) and bacterial organisms in 20 (71.4%) . Bacteria were isolated from mycetomas that had demonstrable A . fumigatus organisms as well as from those that did not have them . In all infected mycetomas Gram negative rods were cultured, in only 4 cases Gram (+) cocci were found . Only in two cases of mycetomas under anaerobic conditions Gram (+) cocci were cultured . In most patients the organism could be demonstrated in sputum as well as in mycetomas . In 18 patients the pathogenic organism disappeared from sputum after removal of mycetomas . Also following removal of aspergillomas a decrease of sputum production was noted and it changed its character from purulent to mucoid. J Intern Med, 1989 Jan, 225(1), 21 - 7 Abnormal proteolysis (DIC)--successful treatment with antithrombin III concentrate and a concentrate containing F XIII and native von Willebrand factor; Delshammar M et al.; Two patients with life-threatening disseminated intravascular coagulation (DIC) syndrome, one caused by Gram-negative bacteria and one by premature separation of the placenta, are described . Specific substitution was given by antithrombin III concentrate and AHF-Kabi, a low purity factor VIII concentrate containing native von Willebrand factor and factor XIII . The treatment quickly returned the extremely low levels of antithrombin III, factor VIII:C, fibrinogen and factor XIII, initially found, to normal, and also returned the multimeric pattern of von Willebrand factor to normal . This resulted in diminished bleeding, enabling surgical treatment of the underlying disease. J Bacteriol, 1989 Jan, 171(1), 308 - 13 Susceptibility of Rhodobacter sphaeroides to beta-lactam antibiotics: isolation and characterization of a periplasmic beta-lactamase (cephalosporinase); Baumann M et al.; Thirteen strains of the gram-negative, facultative phototrophic bacterium Rhodobacter sphaeroides were examined fro susceptibility to beta-lactam antibiotics . All strains were sensitive to the semisynthetic penicillins ampicillin, carbenicillin, oxacillin, cloxacillin, and methicillin, but 10 of the 13 strains were resistant to penicillin G, as well as a number of cephalosporins, such as cephalothin, cephapirin, and cephalosporin C . A beta-lactamase (EC 3.5.2.6) with strong cephalosporinase activity was detected in all of the resistant strains of R . sphaeroides . With strain Y-1 as a model, it was shown that the beta-lactamase was inducible by penicillin G, cephalosporin C, cephalothin, and to some minor extent, cephapirin . The beta-lactamase was located in the periplasmic space, from which it could be extracted by osmotic shock disruption . By using this fraction, the beta-lactamase was purified 34-fold to homogeneity by steps involving batch adsorption to and elution from DEAE-Sephadex A50, chromatography on Q-Sepharose, and preparative polyacrylamide gel electrophoresis . The molecular masses of the native and denatured enzymes were determined to be 38.5 kilodaltons by gel filtration and 40.5 kilodaltons by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively, indicating a monomeric structure . The isoelectric point was estimated to be at pH 4.3 . In Tris hydrochloride buffer, optimum enzyme activity was measured at pH 8.5 . The beta-lactamase showed high activity in the presence of the substrates cephalothin, cephapirin, cephalosporin C, and penicillin G, for which the apparent Km values were 144, 100, 65, and 110 microM, respectively . Cephalexin, cepharidine, and cephaloridine were poor substrates . The beta-lactamase was strongly inhibited by cloxacillin and oxacillin but only slightly inhibited by phenylmethylsulfonyl fluoride or thiol reagents such as iodoacetate and p-chloromercuribenzoate. Microbios, 1989, 59(238), 47 - 63 Microbiological degradation of quinoline by Pseudomonas stutzeri: the coumarin pathway of quinoline catabolism; Shukla OP; A Gram-negative, oxidase positive, polar flagellated rod, characterised as Pseudomonas stutzeri, has been isolated from sewage by enrichment culture on quinoline . The organism utilizes quinoline as the sole source of carbon, nitrogen and energy, and liberates UV absorbing and phenolic metabolites during its growth on quinoline . 2-Hydroxyquinoline, 2,8-dihydroxyquinoline, 8-hydroxycoumarin and 2,3-dihydroxyphenylpropionic acid have been isolated as the transformation products of quinoline by this bacterium . Quinoline, 2-hydroxyquinoline, and 8-hydroxycoumarin were rapidly oxidised by quinoline-adapted cells; 2,3-dihydroxyphenylpropionic acid oxidation was also demonstrated by Warburg respirometry but 2,8-dihydroxyquinoline was not oxidised . A pathway for quinoline catabolism by P . stutzeri and the probable mechanisms for formation of 8-hydroxycoumarin are suggested. Vutr Boles, 1989, 28(1), 60 - 5 {The diagnostic significance of antibodies to lipid A in patients with chronic kidney infections}; Altunkova I et al.; The interest in antilipid A antibodies increases in recent years . By means of an immunoenzyme test developed by the authors the frequency, quantity and characteristic of these antibodies were studied in 46 patients with chronic renal infections and in 36 healthy controls . The frequency of antilipid A antibodies class IgM is 5.7% and of class IgG it is 2.8% in the control group (healthy persons) and it is 41.3% and 19% respectively in the renal patients group . Most frequently their presence and quantity correlate with an active infection or with the convalescent period following antibiotic treatment (62.5%) . This allows the use of antilipid A antibodies as a confirming marker for the presence of a heavy Gram negative infection and as a possible index for evaluation the efficacy of the treatment. Antibiot Khimioter, 1989 Jan, 34(1), 66 - 8 {Antibiotic sensitivity of gram-negative opportunistic bacteria isolated from the feces of newborn infants with intestinal infections}; Litiaeva LA et al.; Antibiotic sensitivity of 136 pure cultures of gram negative aerobic potentially pathogenic bacteria was assayed . The bacteria were isolated from feces of newborns with intestinal infections . It was shown that the majority of the strains were resistant to 4-5 antibiotics widely used in treatment of patients . However, they were sensitive to gentamicin and polymyxin M. Acta Neurochir (Wien), 1989, 97(3-4), 111 - 3 Stunning device as a suicide weapon; Klun B et al.; Thirteen patients who had attempted suicide with the stunning device, were treated over a time span of eleven years . Five of them died . The survivors made a satisfactory recovery with minor neurological sequelae only . The characteristics of this type of brain injury are: high incidence of infection (all developed meningitis and six of them brain abscesses), with prevalence of gram negative animal strains and technical difficulties in removing foreign bodies from the deeper part of the missile track. Z Exp Chir Transplant Kunstliche Organe, 1989, 22(1), 3 - 8 {Endotoxin in experimental mesenteric infarction}; Gogler H; After ligation of the mesenteric arteries an intestinal ischaemia was obtained . Endotoxin was measured from the peritoneal cavity, the portal und the peripheral blood . The limulus test for Endotoxin was used after phenol-water-extraction of the plasma . Microbiological investigations were done simultaneously . The endotoxin concentrations correlated to the clinical state of gram negative sepsis. Sci Prog, 1989, 73(292 Pt 4), 469 - 99 Airborne bacteria and viruses; Cox CS; Coughing, sneezing, talking, bed-making, turning pages of books, etc . all generate microbial aerosols which are carried and dispersed by air movements . Inhalation of these particles may cause allergic responses but whether or not infectious disease ensues depends in part on the viability and infectivity of the inhaled microbes and their landing sites . Desiccation is experienced by all airborne microbes; gram-negative bacteria and lipid-containing viruses demonstrate phase changes in their outer phospholipid bilayer membranes owing to concomitant changes in water content and/or temperature . These changes most likely lead to cross-linking reactions of associated protein moieties principally at mid to high relative humidity (RH) . For lipid-free viruses these reactions of their surface protein moieties occur most rapidly at low RH . Radiation, oxygen, ozone and its reaction products and various pollutants also decrease viability and infectivity through chemical, physical and biological modification to phospholipid, protein and nucleic acid moieties . The extent of damage and the degree of repair together with the efficacy of host defence mechanisms largely controls whether the causative microbes take hold and spread disease via the airborne route . At least indoors, where desiccation is the predominant stress, the general reversibility of membrane-phase changes by vapour-phase rehydration when coupled with efficacious microbial enzymatic repair mechanisms under genetic control, virtually ensures the spread of disease by the aerobiological pathway. Arkh Patol, 1989, 51(12), 74 - 9 {Pathology of the digestive organs and systemic endotoxinemia}; Permiakov NK et al.; The intestine being a natural depot of the gram-negative bacteria, may be the source of the systemic endotoxinemia in various pathologic processes followed by the increase of bacteria death, mucous membrane damage, pancreas endocrine function deficiency, liver barrier function depression, decrease of the portal circulation speed or its shunting . At the same time, the systemic endotoxinemia in itself may be one of the most important etiological factors of the various digestive organs injury . The authors observations and the literature suggest an important role of the endotoxin-positive granulocytes in the pathogenesis of the haemorrhagic infarct of the intestine, mesenteric vein thrombosis, destructive appendicitis and cholecystitis. Scand J Infect Dis, 1989, 21(6), 709 - 15 Early fall of circulating iron and rapid rise of lactoferrin in septicemia and endotoxemia: an early defence mechanism; Gutteberg TJ et al.; Total serum iron, plasma lactoferrin and circulating leukocytes were measured in piglets during the early phase of severe gram-negative septicemia and endotoxemia in 3 experimental settings: intravenous (i.v.) infusion of lipopolysaccharide (LPS) (n = 8), i.v . infusion of live Escherichia coli (n = 7) and intraperitoneal (i.p.) infusion of E . coli (n = 6) . Iron dropped significantly during the first 30 min of LPS infusion from a median of 32 microM to 13.4 microM . A similar decrease in serum iron was demonstrated in the 2 other groups with minimum values at 120 min after the start of E . coli infusion . Plasma levels of lactoferrin increased significantly 120 min after the start of LPS infusion (median 6 mg/l) when compared to preinfusion values (0.25 mg/l) . After i.v . infusion of E . coli a significant rise of plasma lactoferrin was demonstrated already 30 min after bacterial infusion (to 2.1 mg/l) compared to preseptic values (0.8 mg/l) . This increase was accompanied with a significant drop of circulating leukocytes (to 7.3 x 10(9)/l) compared to before the infusion (17 x 10(9)/l) in the pigs given E . coli i.v . After i.p . E . coli infusion no significant change of plasma lactoferrin was observed . The rapid fall of total serum iron seen during endotoxemia and E . coli septicemia may in part be explained by the release of lactoferrin from granulocytes and the clearance of iron-bound lactoferrin in the blood or peritoneal cavity. Eur J Cancer Clin Oncol, 1989, 25 Suppl 2, S25 - 35 Current approaches to management of infections in bone marrow transplants; Winston DJ et al.; Infections continue to be common complications of bone marrow transplantation, but recent advances have improved their outcome . Oral chemoprophylaxis with the fluoroquinolones has reduced gram-negative infections during periods of granulocytopenia, while new triazole drugs show promise for improving antifungal prophylaxis . Similarly, recombinant hematopoietic growth factors may reduce infections by shortening the period of post-transplant granulocytopenia . The efficacy of double beta-lactam antibiotic therapy or monotherapy with imipenem has obviated the need to use aminoglycosides in the empiric treatment of febrile patients receiving cyclosporine or other nephrotoxic agents . Treatment of post-transplant interstitial pneumonia associated with cytomegalovirus (CMV) remains problematic, but recent results using the combination of ganciclovir plus intravenous immune globulin have been favorable . In CMV-seronegative patients, CMV infections and pneumonia can be prevented or modified by using CMV-seronegative blood products and intravenous immune globulin . Intravenous immune globulin also has the additional benefits of modifying graft versus host disease and preventing late bacterial infections after marrow engraftment . In CMV-seropositive patients, prophylactic ganciclovir may prevent CMV reactivation and pneumonia and is the subject of an ongoing controlled clinical trial. Trans R Soc Trop Med Hyg, 1989 Jan-Feb, 83(1), 38 - 41 Antibiotic resistance in the tropics . 1 . The genetics of bacterial ampicillin resistance in tropical areas; Young HK et al.; Ampicillin and its derivatives are the most widely used beta-lactam antibiotics throughout the world . Ampicillin resistance in Gram-negative bacteria is usually manifested by plasmid-encoded beta-lactamases, which hydrolyse the beta-lactam ring of the antibiotic . There are at least 30 different plasmid-encoded beta-lactamases but almost all of them are found very infrequently . The one exception is the TEM-1 beta-lactamase which is found wherever transferable ampicillin resistance emerges and accounts for over 50% of all plasmid encoded ampicillin resistance . In India, the incidence of ampicillin resistance is high (82%) and, amongst Escherichia coli, a significant proportion of the plasmid-encoded beta-lactamases are different from those found in the United Kingdom . Although many Gram-negative species are able to accept the TEM-1 beta-lactamase, certain species have a pre-disposition to their own plasmid beta-lactamase types. Arkh Patol, 1989, 51(5), 77 - 80 {Role of bacterial lipopolysaccharides in provoking immune conflict in the mother-fetus system}; Bandazhevskii IuI; The paper analyzes the role of lipopolysaccharide (LPS) substances of gram-negative bacteria in occurrence of the immune conflict in the mother-fetus system . Proceeding from experimental evidence, the author advances the hypothesis that the immune conflict between the mother and her fetus is induced by bacterial LPS . In so doing, the bacterial agents play a subsidiary role by replacing T helpers when the mother's organism develops an immune response to antigenic determinants of fetal erythrocytes. Immunol Invest, 1989 Jan-May, 18(1-4), 187 - 209 The role of antibody, complement and neutrophils in host defense against Actinobacillus actinomycetemcomitans; Wilson ME et al.; A . actinomycetemcomitans is a facultative Gram-negative coccobacillus which has been implicated in the etiology and pathogenesis of localized juvenile periodontitis and has also been recognized for its potential to cause serious extraoral infections, particularly endocarditis . The polymorphonuclear neutrophil has been suggested to play a key role in host resistance to periodontopathic organisms, as indicated by the association between defective production or function of these phagocytic cells and severe periodontal disease . This association has engendered interest in the study of the interaction between neutrophils and A . actinomycetemcomitans, as well as the role of immunoglobulin and complement in facilitating this interaction . The objective of this review is to summarize current knowledge of the nature and consequences of the interaction between A . actinomycetemcomitans and the host defense triad consisting of neutrophils, complement and immunoglobulin. Clin Exp Pharmacol Physiol, 1989 Jan, 16(1), 1 - 6 Mechanisms of aminoglycoside nephrotoxicity; Bennett WM; Aminoglycosides continue to be widely used for the treatment of serious Gram-negative infections . Ten to fifteen per cent of all courses of therapy are complicated by declines in renal function, despite close monitoring of serum drug levels . The proposed pathogenesis and biochemical mechanisms of renal dysfunction caused by these commonly used therapeutic agents are discussed. Crit Care Clin, 1989 Jan, 5(1), 151 - 5 Review of corticosteroid treatment in sepsis and septic shock: pro or con; Nicholson DP; The use of glucocorticoids in high dosage for the prevention and treatment of sepsis, septic shock, and ARDS has been controversial . The state of our knowledge has been reviewed, particularly to include some recent critical contributions . It is possible that glucocorticoids have a place in the treatment of gram-negative bacteremia, because the evidence in this situation is equivocal . There is no evidence that such therapy is indicated in other septic states, nor is there evidence that glucocorticoids prevent ARDS or improve the outcome . There are inadequate data to state categorically that glucocorticoid therapy in sepsis is harmful. APMIS, 1989 Jan, 97(1), 14 - 22 Cloning and expression of the Legionella micdadei "common antigen" in Escherichia coli; Bangsborg JM et al.; To study individual Legionella antigens, a Legionella micdadei genomic library in Escherichia coli SC181 was established . Partially Sau3A digested L . micdadei DNA fragments (15-25 kilobase pairs (kb} were cloned into the tetracycline resistance gene of the cosmid vector pHC79 . Four thousand ampicillin resistant recombinants were obtained; seven hundred were screened for expression of Legionella antigens in Western blot analysis with a polyspecific E . coli-absorbed anti-L . micdadei rabbit antibody . One of the positive clones expressed a 60 kilodalton (K) antigen, which reacted strongly with a monospecific rabbit antiserum raised against L . micdadei "common antigen" (CA), and an additional 13 K L . micdadei protein . The region encoding these two proteins from the 17 kb recombinant plasmid (pBA 2) was subcloned in pBGS18+ . The DNA sequence of the CA encoding region in the 2.7 kb subcloned fragment will provide important information with respect to genetic vs . antigenic relatedness among Legionellae and other Gram-negative species, as well as to CA structure and possible function. Mayo Clin Proc, 1989 Jan, 64(1), 84 - 94 The first 100 liver transplantations at the Mayo Clinic; Krom RA et al.; Between March 1985 and June 1987, the first 100 liver transplantations at the Mayo Clinic were performed in 83 patients (primarily adults) . The most frequent diagnoses were chronic active hepatitis (in 24 patients), primary sclerosing cholangitis (in 22), and primary biliary cirrhosis (in 20) . The median operating time was 406 minutes, and the median usage of erythrocytes was 13.2 units . A venovenous bypass was used in all patients older than 10 years of age . Hepatic artery thrombosis occurred in 10% of the 100 transplants . A choledochocholedochostomy was done in 58 patients and a choledochojejunostomy in 25 patients . Revision of the biliary anastomosis was necessary in 9 of the 83 patients (11%) . Rejection, diagnosed by clinical and histologic criteria, occurred in 50 patients (60%) and was treated with a corticosteroid bolus, followed by OKT3 (monoclonal antibody) treatment if necessary . Selective bowel decontamination helped prevent infections; only 16 bacteremias occurred, 1 of which was caused by a gram-negative organism . Fungal infections were rare . Cytomegalovirus infection occurred in 47 patients (57%) . Of the 83 patients, 16 required retransplantation, in 11 of whom graft rejection had occurred . One- and 2-year patient survival was 83% and 70%, respectively . Although problems still remain, liver transplantation is a reasonable option for patients with end-stage liver disease. Metab Pediatr Syst Ophthalmol, 1989, 12(4), 110 - 2 Serum secretory IgA levels in patients with Behçet disease; Sunakawa M et al.; Serum IgA, IgC, IgM and secretory IgA levels in patients with Behcet disease were examined . Among them, serum IgA and secretory IgA levels, which increase in mucosal inflammation, were significantly higher than controls (p less than 0.001, p greater than 0.01) . These results indicate that mucosal inflammation, oral and gastrointestinal inflammation, may lead to gram-negative bacterial infection and endotoxin-induced uveitis in patients with Behcet disease. Intensive Care Med, 1989, 15(7), 471 - 4 Calcium free hemodialysis: an effective therapy in hypercalcemic crisis--report of 4 cases; Kaiser W et al.; Hypercalcemic crisis represents a medical emergency . If conservative treatment is ineffective, low calcium bath or zero calcium bath hemodialysis are good alternatives . We report 4 patients treated with calcium free acetate hemodialysis because of hypercalcemic crisis due to breast cancer, hepatocellular carcinoma, cirrhosis of the liver and immobilisation with hydrochlorothiazids' medication . Following 3 h of hemodialysis, serum calcium concentrations fell from a mean value of 3.96 (range 3.53-4.46) mmol/l to 2.71 (2.28-3.12) mmol/l . In 2 patients rapid clinical improvement was achieved and in one oliguric patient diuresis started spontaneously during hemodialysis . One patient died from gram-negative sepsis . In 3 cases the subsequent conservative treatment was sufficient to maintain serum calcium levels within the normal range . Together with the previously reported cases (5 patients treated by hemodialysis with low dialysate calcium and 3 patients by hemodialysis with calcium free dialysate) our experience indicates that hemodialysis is an effective and safe therapy for hypercalcemic crisis. Microbiol Immunol, 1989, 33(11), 941 - 9 Studies on B-cell memory . IV . Effects of lipopolysaccharide on primary and secondary antibody responses to T-independent type-2 (TI-2) antigen in mice; Hosokawa T et al.; Effects of LPS on primary and secondary antibody responses to typical TI-2 antigens were investigated in mice . Simultaneous injection of LPS with a TI-2 antigen showed only little adjuvant effect on the following primary antibody response to the antigen . In contrast, either a single or multiple injections of LPS, prior to the immunization with a TI-2 antigen, significantly augmented the following primary antibody response to the antigen . LPS, however, inhibited the development of B-cell memory to a TI-2 antigen when administered together with the antigen . Moreover, an injection of LPS in mice, which had strong IgM and IgG B-cell memories to a TI-2 antigen, caused disappearance or profound reduction of the memories . The results suggest that LPS produced by gram-negative bacteria exerts inhibitory effects on the development and continuation of B-cell memory to bacterial infections. Arch Microbiol, 1989, 152(6), 556 - 63 Anaerobic degradation of aniline and dihydroxybenzenes by newly isolated sulfate-reducing bacteria and description of Desulfobacterium anilini; Schnell S et al.; A new, rod-shaped, Gram-negative, non-sporing sulfate reducer (strain Ani1) was enriched and isolated from marine sediment with aniline as sole electron donor and carbon source . The strain degraded aniline completely to CO2 and NH3 with stoichiometric reduction of sulfate to sulfide . Strain Ani1 also degraded aminobenzoates and further aromatic and aliphatic compounds . The strain grew in sulfide-reduced mineral medium supplemented only with vitamin B12 and thiamine . Cells contained cytochromes, carbon monoxide dehydrogenase, and sulfite reductase P582, but no desulfoviridin . Strain Ani1 is described as a new species of the genus Desulfobacterium D . anilini . Marine enrichments with the three dihydroxybenzene isomers led to three different strains of sulfate-reducing bacteria; each of them could grow only with the isomer used for enrichment . Two strains isolated with catechol (strain Cat2) or resorcinol (strain Re10) were studied in detail . Both strains oxidized their substrates completely to CO2, and contained cytochromes, carbon monoxide dehydrogenase, and sulfite reductase P 582 . Desulfoviridin was not present . Whereas the rod-shaped catechol oxidizer (strain Cat2) was able to grow on 18 aromatic compounds and several aliphatic substrates, the coccoid resorcinol-degrading bacterium (strain Re10) utilized only resorcinol, 2,4-dihydroxybenzoate and 1,3-cyclohexanedion . These strains could not be affiliated with existing species of sulfate-reducing bacteria . A further coccoid sulfate-reducing bacterium (strain Hy5) was isolated with hydroquinone and identified as a subspecies of Desulfococcus multivorans . Most-probable-number enumerations with catechol, phenol, and resorcinol showed relatively large numbers (10(4)-10(6) per ml) of aryl compound-degrading sulfate reducers in marine sediment samples. Arch Microbiol, 1989, 152(2), 132 - 7 Characterization and localization of phosphatidylglycerophosphate and phosphatidylserine synthases in Rhodobacter sphaeroides; Radcliffe CW et al.; Catalytic properties and membrane associations of the phosphatidylglycerophosphate (PGP) and phosphatidylserine (PS) synthases of Rhodobacter sphaeroides were examined to further characterize sites of phospholipid biosynthesis . In preparations of cytoplasmic membrane (CM) enriched in these activities, apparent Km values of PGP synthase were 90 microM for sn-glycerol-3-phosphate and 60 microM for CDP-diacylglycerol; the apparent Km of PS synthase for L-serine was near 165 microM . Both enzymes required Triton X-100 with optimal PS synthase activity at a detergent/CDP-diacylglycerol (mol/mol) ratio of 7.5:1.0, while for optimal PGP synthase, a range of 10-50:1.0 was observed . Unlike the enzyme in Escherichia coli and several other Gram-negative bacteria, the PS synthase activity had a specific requirement for magnesium and was tightly associated with membranes rather than ribosomes in crude cell extracts . Sedimentation studies suggested that the PGP synthase was distributed uniformly over the CM in both chemoheterotrophically and photoheterotrophically grown cells, while the PS synthase was confined mainly to a vesicular CM fraction . Solubilized PGP synthase activity migrated as a single band with a pI value near 5.5 in a chromato-focusing column and 5.8 on isoelectric focusing; in the latter procedure, the pI was shifted to 5.3 in the presence of CDP-diacylglycerol . The PGP synthase activity gave rise to a single polypeptide band in lithium dodecyl sulfate-polyacrylamide gel electrophoresis at 4 degrees C. Pediatr Pulmonol, 1989, 7(3), 128 - 32 Branhamella catarrhalis bronchopulmonary isolates in PICU patients; Kasian GF et al.; Branhamella catarrhalis is commonly considered a respiratory commensal but has recently been implicated as a pathogen, particularly in adults . Over a 28 month period, B . catarrhalis was isolated from bronchopulmonary secretions of 14 PICU patients with acute respiratory infections . Twelve patients had pneumonia and two had tracheitis . The mean age was 3.5 years . Seven patients had chronic cardiopulmonary disease including two who were immunosuppressed . Three had an acute underlying condition and four had no complicating medical problem . Polymorphs and Gram-negative diplococci on Gram stain were found in respiratory secretions of all patients . Twelve of 14 isolates produced beta-lactamase, and six patients had a second potentially pathogenic bronchopulmonary isolate . All patients were treated for B . catarrhalis infection and none died . When isolated in pure culture from bronchopulmonary secretions in symptomatic patients, B . catarrhalis should be considered a pathogen . When isolated in mixed culture, its pathogenic role is uncertain . We conclude that B . catarrhalis can be a bronchopulmonary pathogen in critically ill children with otherwise normal cardiopulmonary function as well as in those with chronic cardiopulmonary dysfunction . When administering antibiotics the high frequency of beta-lactamase-producing strains must be taken into consideration. DICP, 1989 Jan, 23(1), 33 - 8 Impact of a clinical pharmacokinetic service on patients treated with aminoglycosides for gram-negative infections; Destache CJ et al.; The present study was a retrospective, case-control design aimed at evaluating whether the clinical pharmacokinetic service (CPS) is cost-effective, as assessed by lengths of hospital stay and aminoglycoside therapy, incidence of a decrease in renal function, and time for resolution of infection as determined by vital signs . Forty-six patients were entered into this study, and were matched by defined criteria . The results of the study demonstrated a six-day difference in hospital stay for the CPS group (p less than 0.05) . Length of aminoglycoside therapy was 33 hours shorter for the CPS group . Additionally, the time necessary for resolution of the infection was significantly shorter for this group, as assessed by vital signs returning to normal or baseline . Three patients in each group expired . Two patients in the CPS group and five in the control group developed aminoglycoside-associated increases in serum creatinine . No significant difference was found between the two groups in age, weight, or APACHE II score . Additionally, the two groups were similar with respect to concomitant diseases and concomitant antibiotics used . The approximate cost of the CPS was calculated as $56 per patient . Use of the CPS decreasing hospital stay by six days (mean $1875/patient) would translate to an annual savings of $654,375 in hospital charges, assuming 365 patients received aminoglycoside therapy per year. Ophthal Plast Reconstr Surg, 1989, 5(4), 277 - 80 The anophthalmic socket and the prosthetic eye . A clinical and bacteriologic study; Vasquez RJ et al.; Culture samples were obtained from the anophthalmic sockets of 40 consecutive patients using a prosthetic eye . We found an increased prevalence of bacteria in the conjunctivae of anophthalmic sockets . No significant differences in bacterial flora were observed between symptomatic and asymptomatic patients, suggesting that symptoms of irritation are not usually related to abnormal bacterial flora . Patients who frequently manipulated their prosthesis had a significantly higher proportion of gram-negative bacteria, suggesting that manipulation of the prosthesis should be avoided. Tr Inst Im Pastera, 1989, 66, 104 - 9, 172 {Methods of isolating phospholipids from Coxiella burnetii}; Lukacheva M et al.; Phospholipids are a part of the outside membrane of Coxiella burnetii (or other gram negative bacteria) . It is possible to extract them with a mixture of chloroform-methanol (CM) . For extraction of phospholipids from C . burnetii it is important to take account of temperature, proportion of CM, and reaction time . The best results are obtained at CM 2:1, at boiling point of this mixture and reaction time of 2 hours . Longer reaction time does not influence the reaction, the additional amounts of phospholipids can be extracted only with fresh solvent mixture. Nauchnye Doki Vyss Shkoly Biol Nauki, 1989, (7), 13 - 24 {Proteins--channel formers . II . Porins and proteins of the external mitochondrial membranes}; Krasil'nikov OV; The physical and chemical properties in protein external membranes of gram-negative bacteria and mitochondria are considered . A possible evolutional affinity between these proteins is discussed . The data on peculiarities of their polypeptide chain structure depending on environmental parameters and chemical modification of ionogeneous groups of protein-channel formers as well on a possible structure of water pore in ion channels are presented. Complement Inflamm, 1989, 6(3), 175 - 204 Ba and Bb fragments of factor B activation: fragment production, biological activities, neoepitope expression and quantitation in clinical samples; Kolb WP et al.; Factor B is a centrally important component of the alternative complement pathway . Alternative pathway activation results in factor B cleavage and production of the amino-terminal Ba and the carboxyl-terminal Bb fragments which have molecular weights of approximately 30,000 and 63,000 daltons, respectively . Both Ba and Bb fragments have been reported to express a variety of biological activities in vitro . Thus, binding of Ba and Bb fragments to specific B lymphocyte surface receptors modulates proliferation of prestimulated B cells . In addition, the enzymatically active Bb fragment induces activation and spreading of human and murine macrophages and monocytes as well as regulates C5a des Arg chemotactic activity . The fractional catabolic rate and metabolism of factor B in vivo is similar to that of C3, C4 and C5 complement proteins, which are among the most metabolically active plasma proteins in the circulatory system . Factor B hyperconsumption and increased catabolism, concomitant with factor B fragment production, occurs in a wide variety of diseases, including gram-negative sepsis, autoimmune diseases and burns . Measurement of alternative pathway activation in vivo has been attempted utilized a number of different techniques to quantitate factor B fragments in biological fluids . However, the recent development of enzyme immunoassays (EIA) employing monoclonal antibodies (MoAbs) reactive with factor B fragment neoepitopes provides the best approach currently available for the quantitation of factor B activation fragments . Results obtained using these new MoAb-based EIAs have indicated that factor B fragment concentrations were elevated, as compared with normal donor levels, in EDTA plasma samples obtained from patients with rheumatoid arthritis and systemic lupus erythematosus (SLE) . Plasma concentrations of factor B fragments, especially Ba fragment levels, in these patients showed a positive correlation with disease activity scores . One of the highest disease activity correlations was obtained with Ba fragment measurements in SLE plasma samples . In fact, the results strongly suggested that quantitation of Ba fragment levels in SLE plasma samples more accurately reflected disease activity and was a more sensitive predictor of impending flare in these patients than any other test(s) currently available. An Esp Pediatr, 1988 Dec, 29(6), 435 - 9 {Antibiotic therapy of infected cerebrospinal fluid in hydrocephalus treated by a shunt}; Isla Guerrero A et al.; Nine hydrocephalic shunted children with infected cerebrospinal fluid (CSF) were treated . Ages ranged from 0 to 10 years . Diagnosis was made through clinical symptoms, CSF examination, echographic and computed tomography (CT) . The microorganism which was seen more frequently, was S . epidermidis . This germ was more often found in young children . Treatment of these patients consisted of a systematic change of the shunt which was externally diverted, implantation of a CSF Ommaya reservoir in the lateral ventricle, and intraventricular and systemic administration of antibiotic, were made . Using this protocol CSF sterilization was obtained in all cases, after 5 to 12 days of treatment . CSF shunt infected with S . epidermidis can be effectively cleaned with daily intra-shunt vancomycin, and shunt infected with gram-negative are also cleaned with daily intra-shunt gentamycin. Clin Chest Med, 1988 Dec, 9(4), 623 - 33 Bacterial colonization of the tracheobronchial tree; Salata RA et al.; Bacterial colonization of the tracheobronchial tree is a significant problem in hospitalized and chronically ill individuals . Colonization occurs because of both microbial and host factors, with the foremost mechanism being adherence of pathogenic gram-negative bacteria to mucosal epithelial cells . In the pathogenesis of nosocomial pneumonia, colonization appears to be an essential first step . In the diagnostic approach to hospital-acquired pneumonia, a distinction must be made between colonization and true lower respiratory tract infection . Future approaches to the major problem of colonization with gram-negative bacteria will require a better understanding of pathogenesis and employ strategies that not only focus upon limiting the bacterial pathogens but also attempt to improve upon pulmonary defense mechanisms. Am J Pathol, 1988 Dec, 133(3), 472 - 84 Activation of glomerular mesangial cells by gram-negative bacterial cell wall components; Lovett DH et al.; The cell walls of gram-negative bacteria contain several biologically active components, including lipopolysaccharide (LPS), lipoprotein, and protein 1 . The effects of these individual components and a synthetic analog of lipoprotein, TPP, on several activation parameters of glomerular mesangial cells (MC) were examined . Prostaglandin secretion, synthesis of the autogrowth factor, mesangial interleukin-1 (IL-1), and new synthesis of cellular proteins were assessed as markers of MC activation . All bacterial cell wall components evaluated were active in varying degrees as stimulants of prostaglandin secretion . In general, PGE was the predominant product . TPP and protein 1 also induced substantial secretion of thromboxane . Each cell-wall component was effective in stimulating mesangial IL-1 secretion . The activation of MC was associated with the enhanced synthesis of many cellular proteins in addition to IL-1 . Stimulation by these bacterial components was dependent on the state of the mesangial cell cycle, because nonproliferating cells did not respond to these factors . Activation of MC by gram-negative bacterial cell wall components, with release of vasoactive prostaglandins and peptide mitogens, may be responsible for some of the glomerular hemodynamic alterations and cellular proliferative events associated with sepsis or chronic bacterial infection. Infect Immun, 1988 Dec, 56(12), 3184 - 8 Comparison of type 2 and type 6 fimbriae of Bordetella pertussis by using agglutinating monoclonal antibodies; Li ZM et al.; Two types of fimbriae have been identified on the pathogenic gram-negative organism Bordetella pertussis . Monoclonal antibodies to these fimbriae were produced to better understand the role of fimbriae as serotype-specific agglutinogens and to investigate the antigenic relationship between these fimbriae . Three monoclonal antibodies were identified that specifically agglutinated B . pertussis cells containing the U.S . Reference Factor 2 agglutinogen, and six monoclonal antibodies were produced that agglutinated only those strains containing the U.S . Reference Factor 6 agglutinogen . Indirect immunofluorescence studies and immunogold electron microscopy demonstrated that these monoclonal antibodies bind to an outer membrane component on serotype-specific strains of B . pertussis . All of the monoclonal antibodies reacted with native or partially assembled type-specific fimbriae but not with monomeric fimbrial subunits as indicated by Western blot (immunoblot) analysis . The fimbrial agglutinogens recognized by the monoclonal antibodies were also uniquely reactive with either U.S . Reference Factor 2 or 6 antiserum (Eldering agglutinogen 2 or 6 polyclonal antiserum) in an indirect ELISA . No cross-reactivity of the monoclonal antibodies with the unrelated fimbriae was observed in any of the comparative immunological studies . Some of the monoclonal antibodies agglutinated certain strains of B . bronchiseptica, suggesting that this closely related species can contain antigenically similar fimbriae . These monoclonal antibodies should prove useful for further structural and functional analysis of Bordetella fimbriae and for studies on the role that these antigens play in prevention of infection and disease. J Biol Chem, 1988 Nov 15, 263(32), 16586 - 90 The alpha-lytic protease gene of Lysobacter enzymogenes . The nucleotide sequence predicts a large prepro-peptide with homology to pro-peptides of other chymotrypsin-like enzymes; Epstein DM et al.; alpha-Lytic protease is a 19.8-kDa protein secreted from the Gram-negative bacterium Lysobacter enzymogenes . We have cloned and sequenced the gene for this serine protease . The nucleotide sequence contains an open reading frame which codes for the 198-residue mature enzyme and a potential prepro-peptide, also of 198 residues . The COOH-terminal 49 residues of the pro-peptide are significantly homologous to the propeptides of Streptomyces griseus proteases A and B . We suggest that this pro-peptide region facilitates formation of the active enzyme . A region bridging the NH2-terminal pre- and pro-peptides is homologous to a maize inhibitor of serine proteases . We speculate that this region inhibits enzymatic activity of the prepro-enzyme. Gene, 1988 Nov 15, 71(1), 75 - 84 A broad-host-range shuttle system for gene insertion into the chromosomes of gram-negative bacteria; Barry GF; A deletion derivative of transposon Tn7 containing the Escherichia coli lacZY genes as a selectable marker for insertion of foreign DNA into the chromosomes of soil bacteria was improved to facilitate the cloning of additional genes and their insertion by this element . This report describes a series of plasmid vectors that enable this cloning to be carried out in small, high-copy, narrow host-range plasmids . The final Tn element can then be easily moved (by transposition) without further use of restriction enzymes, to plasmids suitable for delivering it to the bacterial chromosome . The very high specificity for insertion of Tn7 into single locations in bacterial chromosomes has been exploited in the construction of a shuttle system for delivering these Tn7 elements. Anaesth Intensive Care, 1988 Nov, 16(4), 418 - 22 Altered aminoglycoside pharmacokinetics in the critically ill; Beckhouse MJ et al.; We studied prospectively 49 patients being treated in an intensive care unit with aminoglycosides for gram-negative sepsis . Pharmacokinetic data were calculated from three post-dose serum levels using a one-compartment model . Doses required to achieve peak levels between 5 and 10 mg/l with trough levels approximately 1.0 mg/l ranged between 2 and 12 mg/kg per day (mean dose 7 mg/kg per day) . During therapy 60% of the patients had a change in their apparent volume of distribution (Vd) of greater than 20% . These patients were likely to have confirmed infection and to be febrile at the start of treatment . Two to three weeks after discharge ten patients were restudied after a single dose of aminoglycoside . There was a reduction in mean Vd from 0.24 to 0.18 l/kg (P less than 0.02) . Critically ill patients have significantly larger volumes of distribution and may require larger doses per kilogram of body weight of aminoglycoside to achieve therapeutic concentrations . Due to considerable variation in kinetic parameters, the use of standard doses or dosing nomograms is not recommended. J Clin Invest, 1988 Nov, 82(5), 1714 - 21 Bacterial lipopolysaccharide-induced intestinal microvascular lesions leading to acute diarrhea; Mathan VI et al.; Subcutaneous challenge of mice with lipopolysaccharide (LPS) from gram negative bacteria, produced an intestinal microvascular lesion causing fluid exudation into the lumen of the intestine and diarrhea . The microvascular lesion was characterized by endothelial cell damage and microthrombi in the venules and capillaries of the intestinal lamina propria . Marker organisms, given orally to challenged mice, grew in the exuded fluid and could invade the mucosa . Intravenous transfer of postchallenge plasma produced the lesion in normal mice and absorption of such plasma by Sepharose coupled to LPS-antibody abolished this effect . Instillation of large quantities of LPS into the lumen of the intestine produced scattered microvascular lesions, although none of these animals developed diarrhea . Since a similar microvascular lesion has been described in the rectal mucosal lamina propria of adults with acute diarrhea, it is suggested that LPS-induced vascular damage may be a novel mechanism in the pathogenesis of acute diarrhea. Arch Surg, 1988 Nov, 123(11), 1367 - 70 Effect of bacteremia on mortality after thermal injury; Sittig K et al.; Infection is a common cause of death after thermal injury . Therefore, we reviewed the records of 1108 patients who had sustained burns to determine the effect of bacteremia on mortality . Overall, 32 patients died due to sepsis, 26 of whom had positive blood cultures . During this period, 93 patients had 133 bacteremic episodes . The 26 bacteremic patients (28%) who died had larger burns (71% +/- 24%) than the 67 patients who survived (39% +/- 24%), as well as a higher incidence of inhalation injuries . The patients dying of sepsis also were more likely to have polymicrobial bacteremias than the survivors . The distribution of microorganisms causing bacteremia was different between the surviving and nonsurviving patients, with patients sustaining fatal bacteremias having a higher incidence of infections with gram-negative enteric organisms or Pseudomonas . Thus, increased burn size and the presence of an inhalation injury, polymicrobial bacteremia, or gram-negative bacteremia were associated with an increased mortality rate in bacteremic patients with burns. Antimicrob Agents Chemother, 1988 Nov, 32(11), 1684 - 92 RSF1010 and a conjugative plasmid contain sulII, one of two known genes for plasmid-borne sulfonamide resistance dihydropteroate synthase; Radstrom P et al.; The nucleotide sequence of the type II sulfonamide resistance dihydropteroate synthase (sulII) gene was determined . The molecular weight determined by maxicells was 30,000, and the predicted molecular weight for the polypeptide was 28,469 . Comparison with the sulI gene encoded by Tn21 showed 57% DNA similarity . The sulII-encoded polypeptide has 138 of 271 amino acids in common with the polypeptide encoded by sulI . The sulII gene is located on various IncQ (broad-host-range) plasmids and other small nonconjugative resistance plasmids . Detailed restriction maps were constructed to compare the different plasmids in which sulII is found . The large conjugative plasmid pGS05 and the IncQ plasmid RSF1010 contained identical nucleotide sequences for the sulII gene . This type of sulfonamide resistance is very frequently found among gram-negative bacteria because of its efficient spread to various plasmids. Am J Vet Res, 1988 Nov, 49(11), 1950 - 4 Relationship between serologic recognition of Escherichia coli 0111:B4 (J5) and clinical coliform mastitis in cattle; Tyler JW et al.; Serum IgG1 ELISA titers recognizing gram-negative core antigens (Escherichia coli {J5}) were studied at a large dairy in central California . Population mean log10 titer was 2.7357 (equivalent to 1:544) with a SE of 0.03843 . Titers increased with increased lactation number (unstandardized regression coefficient = 0.06733) . Changes in lactation number accounted for only 6.77% of titer variation . Titers less than 1:240 were associated with 5.33 times the risk of clinical coliform mastitis . Also, older cattle were at greater risk to develop clinical coliform mastitis . These factors apparently affect incidence in a nonlinear fashion, with greatly increased risk associated with titers less than 1:240 and with fourth or greater lactations. Mol Gen Mikrobiol Virusol, 1988 Nov, (11), 3 - 12 {Genetic determinants of antibiotic resistance in gram negative bacteria}; Anisimova LA et al.; Modern data on spreading, structural and functional organization and evolution of the genetic determinants of antibiotic resistance in the gram-negative bacteria are reviewed . Some mechanisms for resistance to trimethoprime, sulphonamides, tetracyclines, chloramphenicol aminoglycosides, beta-lactam antibiotics controlled by the plasmid and chromosomal genes are presented . The problem of using the molecular DNA-probes containing the genetical determinants for antibiotic resistance in the practical work of clinical laboratories is discussed. JPEN J Parenter Enteral Nutr, 1988 Nov-Dec, 12(6 Suppl), 43S - 52S Invited comment: lipids and the development of immune dysfunction and infection; Wan JM et al.; Excessive W-6 PUFA metabolism due to high levels of dietary fat intake can encourage infection via prolonged inflammation, enhanced Gram negative survival, reticuloendothelial blockage, immunosuppression, and monokine depression . Lipids can influence host immunity by altering eicosanoid metabolism and membrane structure and function . Further investigations are essential to answer questions regarding the levels and properties of various essential fatty acids in TPN lipid emulsions . Combining the features of LCT in the form of W-3 PUFA (fish oil) and MCT in the form of medium-chain triglyceride in a "structured lipid" may decrease infection and may improve survival rates by producing fewer inflammatory eicosanoids of the two- and four-series, and serving as a more "efficient fuel." The introduction of W-3 polyunsaturated fatty acids into the TPN emulsions as well as into normal diets may provide an important therapeutic advance in the pathogenesis of disease . Such unique antiinflammatory properties of W-3 PUFA require intensive research. Anaesthesist, 1988 Nov, 37(11), 704 - 10 {Prevention of stress hemorrhage in an internal medicine intensive care station: sucralfate versus ranitidine}; Laggner AN et al.; Stress ulcer bleeding is a serious complication of critical illness and is associated with increased morbidity and mortality . For the prophylaxis of stress ulcers, antacids, H2-blockers, or sucralfate are prescribed . While H2-blockers inhibit the secretion of gastric acid, sucralfate appears to provide protection without reducing levels of gastric acid . Inhibition of acid secretion increases gastric pH, allowing bacterial overgrowth of the stomach by Gram negative bacteria, which colonize the pharynx and trachea and increase the risk of nosocomial pneumonia . For this reason, H2 blockers appear disadvantageous, though they offer adequate prophylaxis for stress ulcer bleeding . As it does not increase gastric pH, sucralfate provides adequate protection against Gram negative gastric overgrowth, however its prophylactic efficacy is not generally accepted . Therefore, we compared the H2-blocker ranitidine to sucralfate in the prophylactic treatment of stress ulcer bleeding and studied the incidence of positive bacteriological findings in the blood and bronchial secretions of the two groups . In a randomized study, 84 patients undergoing general intensive care received either ranitidine (6 x 50 to 6 x 100 mg daily i.v.) or sucralfate (6 x 1 g via gastric tube or per os) . Both groups were comparable with respect to age, underlying disorders, and factors predisposing to the development of stress ulcers.(ABSTRACT TRUNCATED AT 250 WORDS) Circ Shock, 1988 Nov, 26(3), 287 - 95 Effects of plasma administration on gram negative shock in granulocytopenic dogs; Crowley JP et al.; We studied the effect of an infusion of citrated plasma on the hemodynamic response and bacterial clearance of acutely septic dogs rendered severely neutropenic with cyclophosphamide . Two hours after a 5 X 10(9)/kg intravenous injection of E . coli, neutropenic control dogs infused with 20 ml/kg of normal saline became hypotensive and remained so for the next 2 hr . In contrast, the infusion of 20 ml/kg of citrated plasma after the initiation of bacteremia was followed by a significant (P less than .01) improvement in the mean arterial pressure (MAP), compared to saline-treated animals . The effect of citrated plasma administration on MAP was unrelated to bacterial clearance or to improvement in cardiac output but was associated with an increase in total peripheral resistance that was sustained for 2 hr after plasma infusion . The infusion of hyperimmune plasma from dogs previously immunized to core glycolipid antigen appeared to have no important advantage over normal plasma . At the doses of plasma that were given, a relatively higher arterial pH was the only observed significant difference (P less than .05) when immune plasma was compared to normal plasma . Resuscitation of septic neutropenic dogs with large volumes of plasma deserves further study with respect to the specific mechanisms responsible for its beneficial effect. Circ Shock, 1988 Nov, 26(3), 227 - 35 Antithrombin-III prevents the lethal effects of Escherichia coli infusion in baboons; Taylor FB Jr et al.; Infusion of Escherichia coli (LD100) was followed by coagulopathic and cell injury responses, cardiovascular collapse, and death in 18 to 32 hr in four out of four baboons . Infusion of AT-III in sufficient amounts to achieve AT-III levels of more than 4 units/ml of plasma before and during the infusion of E . coli reduced the intensity of the coagulopathic and cell injury response and prevented vascular collapse and death in four out of four baboons . Failure to achieve AT-III levels of more than six units/ml at T +60 min during the infusion of E . coli resulted in failure to prevent its lethal effects in three out of three baboons even though levels as high as 10 units/ml were achieved later in the course of the experiment . These studies suggest that thrombin and/or its products can contribute to the inflammatory response to E . coli and that AT-III is of potential value as a prophylactic but not as a therapeutic agent in the treatment of patients at high risk of developing gram negative sepsis. J Surg Res, 1988 Nov, 45(5), 467 - 71 Antiserum to endotoxin in hemorrhagic shock; Pohlson EC et al.; Antiserum to Escherichia coli J5, a mutant endotoxin (LPS) which contains only core determinants, has proven effective in reducing mortality from endotoxic shock due to a wide variety of gram-negative bacteria . Twenty New Zealand white rabbits with coliforms in the gut were subjected to hemorrhagic shock of 36 mm Hg for 3 hr . Treated rabbits were resuscitated with 15 cc of rabbit J5 antiserum (hemagglutinating antibody titer against J5 lipopolysaccharide of 1:1024), remaining shed blood, and lactated Ringer's to achieve a mean arterial blood pressure (MABP) within 20% of baseline . The control group was similarly resuscitated but received 15 cc normal rabbit serum (titer 1:2) . Catheters were removed and rabbits were returned to their cages until death or 5 days of survival . Hemodynamic parameters (heart rate, MABP, cardiac output, and total peripheral resistance) did not differ significantly between groups . However, six treated rabbits survived 5 days (60%) and no control rabbit lived past the third postexperimental day (P less than 0.019) . Our data suggest that systemic endotoxemia may contribute to morbidity and mortality in severe hemorrhagic shock. Presse Med, 1988 Oct 26, 17(37), 1968 - 70 {Antibiotic therapy protocol using ceftazidime 3g/day alone or in combination with vancomycin or amikacin . In febrile episodes in neutropenic patients}; Marie JP et al.; In a preliminary study of 21 febrile episodes in neutropenic patients ceftazidime used as empirical treatment in doses of 3 grams per day succeeded in controlling fever in 74 per cent of the cases . Laboratory studies performed in patients with Gram-negative septicaemia showed clinically effective plasma concentrations of the antibiotic . A trial of ceftazidime (3 g/day) administered alone or combined with amikacin or vancomycin is currently in progress in two medical centres . No statistically significant conclusions could be reached from an intermediate study. J Biol Chem, 1988 Oct 15, 263(29), 14802 - 7 Lipid A binding sites in membranes of macrophage tumor cells; Hampton RY et al.; Lipopolysaccharide affects a variety of eukaryotic cells and mammalian organisms . These actions are involved in the pathogenesis of Gram-negative septicemia . Many of the actions of lipopolysaccharide are believed to be caused by its active moiety, lipid A . Our laboratory has previously identified a bioactive lipid A precursor, termed lipid IVA (Raetz, C . R . H., Purcell, S., Meyer, M . V., Qureshi, N., and Takayama, K . (1985) J . Biol . Chem . 260, 16080-16888), which can be labeled with 32P of high specific activity and purified . In this work we have used the labeled probe, 4'-32P-lipid IVA, to develop a novel assay for the specific binding of lipid IVA to whole cells . We have also demonstrated its use in a ligand blotting assay of immobilized cellular proteins . Using the whole cell assay, we show that 4'-32P-lipid IVA specifically binds to RAW 264.7 macrophage-like cultured cells . The binding is saturable, is inhibited with excess unlabeled lipid IVA, and is proteinase K-sensitive . It displays cellular and pharmacological specificity . Using the ligand blotting assay, we show that several RAW 264.7 cell proteins can bind 4'-32P-lipid IVA . The two principal binding proteins have Mr values of 31 and 95 kDa, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Fractionation studies indicate that the 31-kDa protein is enriched in the nuclear fraction and may be a histone, whereas the 95-kDa protein is enriched in the membrane fraction . The binding assays that we have developed should lead to a clearer understanding of lipid A/animal cell interactions. Cancer Res, 1988 Oct 15, 48(20), 5766 - 9 Production of tumor necrosis factor in nude mice by muramyl peptides associated with bacterial vaccines; Noso Y et al.; Gram-negative vaccines can elicit the production of tumor necrosis factor (TNF) in mice primed by muramyl dipeptide (MDP) or by its lipophilic derivative MDP-dipalmitoyl glycerol (MDP-GDP) . In mice pretreated with MDP and particularly with MDP-GDP, Bordetella pertussis vaccine was shown to be more effective than typhoid vaccine . The time course of TNF production in the blood did not indicate any difference between the effect of MDP or of MDP-GDP . In both cases the cytotoxic activity reached maximal levels by 2 h after injection of the bacterial preparations and returned to normal values between 3 and 5 h after the challenge . In nude mice, high titers of circulating TNF were also produced by combined treatment with MDP-GDP and bacterial vaccine . Moreover, in tumor-bearing mice the association of MDP or of MDP-GDP to a bacterial vaccine induced a strong hemorrhagic necrosis, whereas each treatment alone was inactive . It was also found that mice were less sick when they were primed with MDP-GDP than with MDP, and when TNF was elicited by B . pertussis instead of lipopolysaccharide . Moreover, nude mice appeared more resistant to shock and to hemoconcentration than normal mice. J Biol Chem, 1988 Oct 15, 263(29), 14684 - 9 Properties of Escherichia coli mutants lacking membrane-derived oligosaccharides; Fiedler W et al.; Membrane-derived oligosaccharides (MDO) consist of branched substituted beta-glucan chains and are present in the periplasmic space of Escherichia coli and other gram-negative bacteria . A procedure for the isolation of mutants defective in MDO synthesis is described . Their phenotype was compared with a mdoA mutant previously identified, and they are mapped in the mdoA region . Mutants lacking MDO showed imparied chemotaxis on tryptone swarm plates, a reduced number of flagella, and an enhanced expression of the OmpC porin . Revertants able to form swarm rings again had regained the ability to synthesize MDO and showed the wild-type porin pattern . A second group of chemotactic revertants were mutated in the ompB gene region involved in osmoregulation, and they were still devoid of MDO . These findings provide evidence for a link between MDO biosynthesis and other functions of E . coli related to its adaptation to the environment. Gene, 1988 Oct 15, 70(1), 191 - 7 Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria; Keen NT et al.; Improved broad-host-range plasmid vectors were constructed based on existing plasmids RSF1010 and RK404 . The new plasmids pDSK509, pDSK519, and pRK415, have several additional cloning sites and improved antibiotic-resistance genes which facilitate subcloning and mobilization into various Gram-negative bacteria . Several new polylinker sites were added to the Escherichia coli plasmids pUC118 and pUC119, resulting in the new plasmids, pUC128 and pUC129 . These plasmids facilitate the transfer of cloned DNA fragments to the broad-host-range vectors . Finally, the broad-host-range cosmid cloning vector pLAFR3 was improved by the addition of a double cos casette to generate the new plasmid, pLAFR5 . This latter cosmid simplifies vector preparation and has permitted the rapid cloning of genomic DNA fragments generated with Sau3A . The resulting clones may be introduced into other Gram-negative bacteria by conjugation. Gene, 1988 Oct 15, 70(1), 181 - 9 Versatility of a vector for expressing foreign polypeptides at the surface of gram-negative bacteria; Charbit A et al.; A wide variety of peptides in terms of length and sequence can be expressed at the surface of the bacterium Escherichia coli by genetic insertion into a 'permissive' site of the outer membrane protein LamB, used as a carrier . The resulting hybrid proteins essentially keep their biological activities with inserts of up to about 60 amino acid residues, and of a large range of predicted structures or hydrophobicities . This reflects a remarkable flexibility in the organization of the protein, but also in the export machinery . The method used to select such a permissive site is quite general and its potential to generate applications, including a versatile type of live bacterial vaccine, are discussed. Biochemistry, 1988 Oct 4, 27(20), 7919 - 24 Fluorescence measurement of the kinetics of DNA injection by bacteriophage lambda into liposomes; Novick SL et al.; Bacteriophage lambda attaches to Gram-negative bacteria using the outer membrane protein LamB as its receptor . Subsequently, DNA is injected by the bacteriophage into the host cell for replication and expression . The mechanism of DNA injection, however, is poorly understood . In order to begin to characterize DNA injection, a quantitative kinetic assay to detect injection into reconstituted LamB liposomes is described . The technique involves monitoring the increase in fluorescence of liposome-encapsulated ethidium bromide, which occurs as DNA enters the aqueous compartment of the vesicles . The data indicate that injection is several times faster than indicated by earlier studies and is complete within 1 min . Such assays which allow direct observation of this process are necessary first steps toward a mechanistic understanding. Infect Immun, 1988 Oct, 56(10), 2552 - 7 Killing of Actinobacillus actinomycetemcomitans by human lactoferrin; Kalmar JR et al.; Actinobacillus actinomycetemcomitans is a fastidious, facultative gram-negative rod associated with endocarditis, certain forms of periodontal disease, and other focal infections . Human neutrophils have demonstrated bactericidal activity against A . actinomycetemcomitans, and much of the oxygen-dependent killing has been attributed to the myeloperoxidase-H2O2-halide system . However, the contribution of other neutrophil components to killing activity is obscure . Lactoferrin, an iron-binding glycoprotein, is a major constituent of neutrophil-specific granules and is also found in mucosal secretions . In this report, we show that human lactoferrin is bactericidal for A . actinomycetemcomitans . Killing activity required an unsaturated (iron- and anion-free) molecule that produced a 2-log decrease in viability within 120 min at 37 degrees C at a concentration of 1.9 microM . Besides exhibiting concentration dependence, killing kinetics were affected by minor variations in temperature and pH . Magnesium, a divalent cation thought to stabilize lipopolysaccharide interactions on the surface of gram-negative organisms, enhanced lactoferrin killing of A . actinomycetemcomitans, while other cations, such as potassium and calcium, had no effect . Our data suggest that lactoferrin contributes to killing of A . actinomycetemcomitans by human neutrophils and that it may also play a significant role in innate secretory defense against this potential periodontopathogen. Infect Immun, 1988 Oct, 56(10), 2650 - 7 Recombinant interleukin-1 alpha and recombinant tumor necrosis factor alpha synergize in vivo to induce early endotoxin tolerance and associated hematopoietic changes; Vogel SN et al.; Endotoxin, the lipopolysaccharide (LPS) derived from gram-negative bacteria, invokes a wide range of responses in susceptible hosts . It is known that virtually all responses to LPS are mediated by the action of macrophage-derived cytokines (such as interleukin-1 {IL-1}, tumor necrosis factor {TNF}, and others) which are produced principally by macrophages and maximally within several hours of LPS administration . One manifestation of LPS administration which is not well understood is the phenomenon of "early endotoxin tolerance." In response to a single sublethal injection of LPS, experimental animals become refractory to challenge with a homologous or heterologous LPS preparation 3 to 4 days later . Animals rendered tolerant exhibit mitigated toxicity and a reduced capacity to produce circulating cytokines (i.e., colony-stimulating factor or interferon) in response to the challenge LPS injection . Previous studies have also shown that this state of transient, acquired hyporesponsiveness to LPS is accompanied by a marked increase in the size of cells in the bone marrow which are enriched in numbers of macrophage progenitors . In this study, we examined the capacity of recombinant IL-1 or recombinant TNF or both to induce early endotoxin tolerance and its associated hematopoietic changes . Neither cytokine alone was able to mimic LPS for induction of tolerance . Combined administration of recombinant IL-1 and recombinant TNF doses which were not toxic when administered individually led to synergistic toxicity (as assessed by death or weight loss) . However, within a nontoxic range, the two cytokines synergized to induce a significant reduction in the capacity to produce colony-stimulating factor in response to LPS, as well as the characteristic increase in bone marrow cell size and macrophage progenitors shown previously to be associated with LPS-induced tolerance. Ann Ophthalmol, 1988 Oct, 20(10), 383 - 4 Lupus optic neuritis with negative serology; Deutsch TA et al.; A 56-year-old woman with a 12-year history of systemic lupus erythematosus presented with severe optic-disc swelling and blepharitis . At the same time, she developed acute pancreatitis and ultimately died of gram-negative sepsis . Although it appeared that the ocular and systemic disorders were manifestations of lupus, her serum antinuclear antibody and complement levels remained normal throughout her hospital course . Optic neuritis may be secondary to lupus, but the diagnosis is difficult to make when the serology is negative. J Clin Microbiol, 1988 Oct, 26(10), 2206 - 8 Antithrombin III in the diagnosis of infection in febrile neutropenic patients; Kibbler CC et al.; Antithrombin III levels in plasma were measured in 29 febrile neutropenic patients (31 episodes) . They were lowest on day 2 in 62.5% of febrile patients and then increased or were normal in 81.3% . Low levels were present on day 2 in only 28.6% of patients with gram-negative infections, as compared with 33% of patients with doubtful infections. J Clin Microbiol, 1988 Oct, 26(10), 2096 - 100 Comparative evaluation of selective media for isolation of Pseudomonas cepacia from cystic fibrosis patients and environmental sources; Carson LA et al.; Pseudomonas cepacia has recently emerged as an important pathogen affecting cystic fibrosis (CF) patients . We evaluated three selective media to assess their comparative potential for identification of patients colonized with P . cepacia and for efficacy of detection of P . cepacia in environmental fluids . Test organisms included P . cepacia isolates from CF patients (10 each from two CF centers), non-CF patients (10 isolates), and environmental sources (10 isolates) . Microbiologic assays were done by the membrane filter procedure; filters were placed on P . cepacia medium (PCM), OFPBL, TB-T, MacConkey agar (MAC), and blood agar (BA) or Standard Methods (SM) sugar, and colonies were counted after incubation at 30 or 35 degrees C for 72 h . Mean recovery efficiencies (MREs) (mean CFU/ml on selective media compared with CFU/ml on BA controls) for environmental and non-CF P . cepacia and patient isolates from one CF center showed a rank order of PCM greater than OFPBL greater than TB-T; for isolates from a second CF center, a rank order of PCM greater than TB-T greater than OFPBL was obtained . MREs for CF center isolates were generally lower than for non-CF patients or environmental isolates on P . cepacia-selective media . With MAC, the MREs for each group of CF isolates were extremely low (14 and 2%) compared with those for non-CF patient (47%) or environmental (84%) isolates . In laboratory and field studies, PCM and OFPBL showed good selectivity against bacteria commonly associated with CF patient respiratory secretions . These findings show that selective media should be used in clinical settings where P . cepacia is sought . With environmental fluids from CF centers, P . cepacia-selective media showed low selectivity against a variety of gram-negative water bacteria and appeared to afford little advantage over SM agar for isolating P . cepacia from environmental samples. Chemioterapia, 1988 Oct, 7(5), 309 - 12 Antibiotic susceptibility of bacteria isolated from active, therapy-resistant periodontal sites; De Luca M; Periodontally affected sites may show resistance to mechanical therapy associated with conventional treatment . This periodontal situation is the result of the cooperation of bacteria, and in some cases the development of superinfections due to opportunistic bacteria is possible . These situations need particular attention in the choice of antibiotics . From some sites, bacteria not commonly found as periodontal pathogens may be isolated; they are generally resistant to antibiotics administered in periodontal therapy . Attention should be used particularly in cases where lesions most resemble those characteristic of the action of gram-negative bacteria . According to results of the present research these cases should be managed with antibiotic combinations and conventional mechanical therapy. J Infect Dis, 1988 Oct, 158(4), 761 - 5 Comparison of the outer membrane proteins of 50 strains of Branhamella catarrhalis; Bartos LC et al.; Branhamella catarrhalis colonizes the respiratory tract of humans and commonly causes otitis media in children and respiratory infections in adults with chronic lung disease . In view of the emergence of this organism as an important human pathogen, we used sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to examine the outer membrane proteins (OMPs) of 50 strains of B . catarrhalis . OMPs were isolated from broth culture supernatants . Typical of other gram-negative bacteria, eight proteins ranging in molecular weight (MW) from approximately 98,000 to 21,000 daltons were revealed by SDS-PAGE; these proteins were designated OMP A-OMP H . Of the OMPs identified, four were heat modifiable (C, D, E, and H) . The 50 strains were obtained from diverse geographic and clinical sources . The OMP patterns were strikingly homogeneous; there was minimal variability in the MW of OMPs between strains . Future studies should establish whether the similarity in MWs of OMPs is paralleled by their antigenic characteristics. J Bacteriol, 1988 Oct, 170(10), 4493 - 500 Expression of the gene encoding the 17-kilodalton antigen from Rickettsia rickettsii: transcription and posttranslational modification; Anderson BE et al.; Recently, we reported the molecular cloning and nucleotide sequence analysis of a gene from Rickettsia rickettsii that codes for a 17-kilodalton antigen (17K antigen) and is preceded by sequences closely resembling the -10 and -35 consensus sequences for recognition by Escherichia coli RNA polymerase (Anderson et al., J . Bacteriol . 169:2385-2390, 1987) . Experiments described in this report indicate that the start sites for initiating transcription of the 17K antigen gene are identical in the E . coli clone and in intact R . rickettsii . In each case, initiation was shown to begin 9 bases downstream of the presumed Pribnow box sequence (TATACT) . A 169-base-pair fragment containing the promoter sequence initiated transcription in both directions when cloned into an E . coli promoter probe vector . The rickettsial fragment was found to contain sequences identical to the -10 region (but not the -35 region) of the E . coli promoter consensus sequence directed away from the 17K antigen gene . The amino-terminal portion (residues 17 to 20) of the deduced amino acid sequence for the 17K antigen contained the tetrapeptide Leu-Gln-Ala-Cys, a sequence that conforms favorably to those described for lipid modification and cleavage by lipoprotein signal peptidase II . The 17K antigen produced by the E . coli clone was shown to be labeled with {3H}palmitate and {3H}glycerol, indicative of lipid modification . In vitro mutagenesis designed to alter the cysteine at residue 20 to a glycine abolished incorporation of {3H}palmitate, suggesting that posttranslational modification occurs via a mechanism similar to that described for other gram-negative bacterial lipoproteins. Chemioterapia, 1988 Oct, 7(5), 327 - 9 Chemoprophylaxis of bacterial infections in granulocytopenic cancer patients using norfloxacin; Casali A et al.; Sixty-five cancer patients pretreated with chemo or radiotherapy, with granulocytopenia less than 1000/mm3 and without fever, were entered into this study: 30 of them were submitted to prophylaxis with norfloxacin while the remaining 35 patients were used as a control group . 20% of the treated subjects versus 68.6% of the controls presented a subsequent infection (P less than 0.001), the lung representing the most frequent site of the infectious disease in both groups (3/6 and 14/24 respectively) . These data strongly suggest the use of norfloxacin as an effective prophylactic drug in nonfebrile, granulocytopenic cancer patients, especially as far as gram-negative infections are concerned . Because of the high prevalence of lung cancer in the patients of our study, and a related prevalence of lung infections, at the present time, a wider use of this antibiotic in every kind of solid tumor cannot be generalized. Ann Surg, 1988 Oct, 208(4), 532 - 40 Intestinal gram-negative bacterial overgrowth in vivo augments the in vitro response of Kupffer cells to endotoxin; Billiar TR et al.; A number of disease states and therapeutic maneuvers common to surgical patients can result in changes in the intestinal flora, permitting bacterial overgrowth and translocation of bacteria to gut lymphoid tissue . It is possible that these changes in gut flora increase portal levels of several factors that are capable of altering macrophage activation state, including endotoxin, lymphokines, and eicosanoids . Since Kupffer cells are directly exposed to gut factors via the portal circulation, changes in intestinal flora may influence Kupffer cell responses . Using germfree rats, it has previously been shown that the presence of gut bacterial flora is important in inducing Kupffer cells to respond to endotoxin, and that an overgrowth of gram-negative bacteria can further augment Kupffer cell responses, supporting the above-mentioned hypothesis . The current set of experiments examines how intestinal gram-negative bacterial overgrowth in normal adult rats effects the response of Kupffer cells to septic stimuli . Kupffer cells were obtained from conventional rats with induced intestinal overgrowth with Escherichia coli C25 for 2 or 7 days . After 2 days of overgrowth, Kupffer cells were only slightly less responsive to lipopolysaccharide (LPS) than control Kupffer cells . However, after 7 days of overgrowth, when placed in coculture with normal hepatocytes, Kupffer cells were significantly more responsive to LPS (p less than 0.001), inducing a greater degree of suppression in hepatocyte protein synthesis at lower LPS concentrations . When cultured alone, Kupffer cells from these animals also produced more interleukin-1 (p less than 0.002) and prostaglandin E2 (PGE2) (p less than 0.009) in response to LPS . These results show that intestinal gram-negative bacterial overgrowth in conventional rats can have direct influences on the response of hepatic macrophages to septic stimuli, and provides further support to the hypothesis that imbalances in the intestinal flora can effect the responses of immune cells in other sites of the body. Gene, 1988 Sep 30, 69(2), 237 - 44 Molecular analysis of the gene encoding alpha-lytic protease: evidence for a preproenzyme; Silen JL et al.; A 1.7-kb EcoRI fragment containing the structural gene for alpha-lytic protease has been cloned from Lysobacter enzymogenes 495 chromosomal DNA: the first example of a gene cloned from this organism . The protein sequence deduced from the nucleotide sequence encoding this serine protease matches the published amino acid sequence {Olson et al., Nature 228 (1970) 438-442} precisely . Sequence analysis and S1 mapping indicate that, like subtilisin {e.g., Wells et al., Nucleic Acids Res . 11 (1983) 7911-7925} alpha-lytic protease is synthesized as a pre-pro protein (41 kDa) that is subsequently processed to its mature extracellular form (20 kDa) . This first finding of a large N-terminal protease precursor in a Gram-negative bacterial protease strengthens the hypothesis that large precursors may be a general property of extracellular bacterial proteases, and suggests that the N- or C-terminal location of the precursor segment may be significant. J Biol Chem, 1988 Sep 25, 263(27), 13479 - 81 A family of lipopolysaccharide binding proteins involved in responses to gram-negative sepsis; Tobias PS et al.; The lipopolysaccharides (LPS) of Gram-negative bacteria initiate potentially fatal processes in many host organisms . Recently published amino acid sequence data suggest that there is a family of LPS binding proteins that may participate in the host response to Gram-negative bacteremia . The first two members of the family to be identified are an LPS binding protein present in serum after an acute phase response in humans, mice, rabbits, and rats and a bactericidal/permeability increasing protein present in the primary granules of human and rabbit neutrophils . LPS binding protein and bactericidal/permeability increasing protein share an ability to bind to LPS, have homologous NH2-terminal amino acid sequences, and are immunologically cross-reactive . Nevertheless, these two molecules differ in their effects on LPS and Gram-negative bacteria, in their sites of biosynthesis, and localization in vivo. J Mol Biol, 1988 Sep 5, 203(1), 275 - 8 Naturally crystalline porin in the outer membrane of Bordetella pertussis; Kessel M et al.; The Gram-negative bacterium Bordetella pertussis is the agent responsible for whooping-cough, and much interest has focused on the functions, structures and immunological properties of the molecules exposed at its outer surface . We have found by electron microscopy that cells of two strains of B . pertussis are covered with a crystalline surface lattice . This lattice is not an extrinsic layer of high molecular weight glycoproteins, such as occur on many other bacteria, but is a natural crystal of an intrinsic membrane protein of 40,000 Mr . This molecule has been shown to be an anion-selective member of an extensive family of proteins ("porins") that render Gram-negative outer membranes permeable to solutes of up to approximately 650 Mr . Computer image processing reveals a trimeric channel-like structure that closely resembles other porins visualized in artificial arrays after treatment with detergents, but in a novel (p2) crystal form . This correlation provides a "missing link" between earlier structural studies based on artificial arrays of porins (of undefined physiological status), and membrane-permeabilization experiments with solubilized porins (in undefined structural states) . For the strains characterized so far, crystallinity of the porin surface lattice shows an intriguing correlation with nonpathogenicity. Biotechniques, 1988 Sep, 6(8), 752 - 60 A plasmid-based method to quantitate homologous recombination frequencies in gram-negative bacteria; Xu BW et al.; A method is described which enables quantitative evaluation of the ability of gram-negative bacterial cells to perform homologous recombination between DNA molecules . This method is particularly useful in cases where the stringency of rec mutations is to be determined . The procedure is based on a wide-host-range vector (pRK404) in which two unequally truncated and overlapping fragments of the neo gene were cloned . When introduced into gram-negative bacteria either by transformation or by conjugation, molecules of this plasmid, pBX404-7, undergo unequal crossing-over leading to the restoration of a functional neo gene . The stringency of putative rec mutations can thus be determined by measuring the frequency at which kanamycin-resistant colonies appear in bacterial strains harboring pBX404-7. Int J Clin Pharmacol Ther Toxicol, 1988 Sep, 26(9), 432 - 5 Some questions of aminoglycoside therapy in the intensive care unit; Csapo K et al.; The pharmacokinetics of tobramycin and gentamycin was studied in patients of the intensive care unit . The "blood-bronchus barrier" was studied by determining the serum and sputum concentrations of the two drugs . At systemic administration, both aminoglycosides passed into the bronchial secretion and when they were inhaled, they appeared in the serum . By culturing the sputum samples of the patients of the intensive care unit, a high ratio of gram-negative pathogens was found . Bacteria were detected in some cases on the objects surrounding the patients, too. Vet Microbiol, 1988 Sep, 18(1), 27 - 39 Development and evaluation of an enzyme-linked immunosorbent assay for endotoxin in milk; Mohammed AH et al.; A double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the detection of endotoxin in milk samples . Bovine and rabbit antisera raised in response to vaccination with the J5 mutant of Escherichia coli 0111:B4 were used . Antiserum to this mutant has been shown to be cross-reactive with endotoxin from other gram-negative organisms . Known quantities of endotoxin were added to milk samples to generate a standard curve . Acid treatment of whole milk enhanced the detection of endotoxin as compared to untreated whole milk, skim milk and chloroform-treated milk . Milk samples from experimentally induced mastitic cows were then assayed for endotoxin content . Recovery of endotoxin, as measured by ELISA, positively correlated with the amount of endotoxin infused and the time post-infusion of sampling . However, when endotoxin from these samples was quantitated using the Limulus Amebocyte Lysate (LAL) assay, readings tended to increase, suggesting false-positive reactions with the LAL assay . Milk samples from cases of clinical mastitis were assayed by ELISA with 64% of these showing measurable levels of endotoxin . While further studies of this assay are needed, refinements may produce an assay important for clinical applications. Oral Surg Oral Med Oral Pathol, 1988 Sep, 66(3), 287 - 9 Metronidazole in the management of anaerobic neck infection in acute leukemia; Barrett AP; A case involving a rapidly progressive neck infection in a severely neutropenic patient with acute leukemia is reported . Resolution followed the addition of metronidazole to an existing standard piperacillin/gentamicin combination, which was used primarily to cover potential gram-negative bacillary pathogens . This indicated the importance of considering extension of anaerobic cover in such infections. Circ Shock, 1988 Sep, 26(1), 15 - 26 Synergistic protection from lung damage by combining antithrombin-III and alpha 1-proteinase inhibitor in the E . coli endotoxemic sheep pulmonary dysfunction model; Redens TB et al.; Septicemic/endotoxic-induced adult respiratory distress syndrome (ARDS) remains a major clinical problem . The present study was to determine in the E . coli endotoxemic sheep ARDS model the efficacy of combination prophylaxis with antithrombin-III (AT-III) and alpha 1-proteinase inhibitor (alpha 1-PI) . We reasoned that 1) AT-III supplementation would ameliorate the endotoxin-induced coagulopathy, 2) alpha 1-PI supplementation would attenuate pulmonary damage caused by neutrophil elastase and inactivation of AT-III by neutrophil elastase, and 3) the therapeutic effects of this combination would be additive or synergistic . The typical increases in lung lymph flow microvascular permeability to protein, transvascular protein flow and transvascular protein clearance, and decrease in systemic arterial PO2 were prevented or significantly attenuated during 5 hours of endotoxemia by the AT-III/alpha 1-PI combination pretreatment . Limited efficacy was observed with AT-III pretreatment, and none was seen with alpha 1-PI alone . Results of this study demonstrate that combining AT-III and alpha 1-PI prophylaxis prevents or attenuates indices of ARDS during gram-negative endotoxemia and that this efficacy is due to a statistically significant synergism between AT-III and alpha 1-PI. Appl Environ Microbiol, 1988 Sep, 54(9), 2203 - 7 Degradation of bromacil by a Pseudomonas sp; Chaudhry GR et al.; A gram-negative rod, identified as a Pseudomonas sp., was isolated from soil by using bromacil as the sole source of carbon and energy . During growth on bromacil or 5-bromouracil, almost stoichiometric amounts of bromide were released . The bacterium was shown to harbor two plasmids approximately 60 and 100 kilobases in size . They appeared to be associated with the ability to utilize bromacil as a sole source of carbon and also with resistance to ampicillin . This microorganism also showed the potential to decontaminate soil samples fortified with bromacil under laboratory conditions. Rev Infect Dis, 1988 Sep-Oct, 10(5), 998 - 1004 Bone and joint infections caused by Kingella kingae: six cases and review of the literature; de Groot R et al.; Six cases of Kingella kingae arthritis, osteomyelitis, and diskitis were studied, and data were reviewed from an additional 23 cases in the English-language literature . K . kingae is a slow-growing, fastidious, gram-negative microorganism that colonizes mucous membranes of the upper respiratory tract . Infections were predominantly seen in infants and young children (86% of cases) and were preceded by an upper respiratory tract infection in 31% of patients . Low-grade fever (38 degrees C-39 degrees C) and pain or swelling involving the affected limb developed insidiously . However, 76% of the infections were diagnosed within 1 week after the onset of symptoms . The knee was involved in 47% and the hip in 33% of cases of arthritis . Osteomyelitis mainly involved the femur (36%); four cases of osteomyelitis (29%) were diagnosed as diskitis . The erythrocyte sedimentation rate was greater than 20 mm/h in all patients . Gram stains of aspiration fluid were positive in 19% of cases, whereas blood cultures yielded growth in only 5% . Radiographic signs indicating the presence of osteomyelitis were observed in 93% of patients . All patients recovered completely . K . kingae is an important causative agent in indolent bone, joint, and intervertebral disk infections. J Clin Invest, 1988 Sep, 82(3), 964 - 71 Modulation of human platelet protein kinase C by endotoxic lipid A; Grabarek J et al.; Lipid A is the toxic principle of lipopolysaccharide of gram-negative bacteria, which causes a spectrum of changes in blood cells and vascular cells . We now report that human platelets are directly stimulated by endotoxic lipid A that activates protein kinase C . Rapid phosphorylation of a human platelet protein of Mr 47,000, a marker of protein kinase C activation, accompanies secretion of {14C}serotonin and aggregation triggered by endotoxic lipid A . These events are time and concentration dependent, with phosphorylation reaching maximum in 2 min and the concentration of lipid A causing a 50% effect (EC50) between 12 and 15 microM . Phospholipase C activation in lipid A-stimulated platelets was not observed as judged by a lack of generation of {3H}diacylglycerol in {3H}arachidonic acid-labeled platelets and a lack of generation of {32P}-phosphatidic acid in 32PO4-labeled platelets . Lipid A did not induce formation of TXA2 as measured by radioimmunoassay for TXB2 . The stimulation of human platelets and activation of protein kinase C by endotoxic lipid A was blocked by lipid X, a structural precursor of lipid A . Lipid X also blocked the stimulation of human platelets by phorbol 12-myristate 13-acetate, suggesting that lipid A, lipid X and phorbol ester share reactive site(s) on the human platelet membrane . Although lipid X inhibited thrombin-induced phosphorylation of P47 it did not suppress secretion of {14C}serotonin, indicating the role of protein kinase C-independent pathways in platelet stimulation by thrombin . The inhibitory effect of lipid X did not involve generation of cyclic AMP in human platelet membrane preparations . These results indicate that human platelets are stimulated by endotoxic lipid A, a naturally occurring biologic modifier of protein kinase C . Due to the widespread presence of this enzyme in blood cells, vascular cells, and neurons, its modulation by lipid A may represent a significant mechanism underlying hematologic and circulatory derangements observed in endotoxic shock in humans. Clin Chest Med, 1988 Sep, 9(3), 449 - 57 Bacterial and fungal pneumonias; Fels AO; Bacterial pneumonias occur with increased frequency and can be associated with increased morbidity in the HIV-infected population compared with normals . The pathogens that most frequently cause community-acquired pneumonias are S . pneumoniae, H . influenzae, and occasionally S . aureus . These pneumonias usually respond to appropriate antibiotic therapy; however, patients diagnosed with bacterial pneumonias are at increased risk for subsequent episodes . Nosocomial pneumonias, by contrast, are usually caused by gram-negative organisms and have a high mortality . Fungal pneumonias also have an increased incidence in AIDS patients, and usually occur in the setting of disseminated disease . Infections caused by C . neoformans, H . capsulatum, and C . immitis often recur despite a good initial response to amphotericin B . Maintenance therapy with an antifungal agent is therefore recommended. Crit Care Med, 1988 Sep, 16(9), 848 - 51 Effect of scavengers of oxygen-derived free radicals on mortality in endotoxin-challenged mice; Broner CW et al.; Oxygen-derived free radicals have been implicated as mediators of cellular injury in several model systems . Recently, a role for free radicals has been proposed in the mortality associated with Gram-negative bacterial sepsis . To determine if pretreatment with free radical scavengers can prevent endotoxin-induced mortality, mice rendered sensitive to endotoxin with actinomycin D were treated with either superoxide dismutase (SOD), N-acetylcysteine (NAC) or saline and were then challenged with a dose of endotoxin calculated to cause a mortality of greater than 80% . Mortality was assessed at 12-h intervals after challenge . Increased survival was seen in the SOD-treated group compared to the control group (p less than or equal to .05) . In contrast, survival in mice treated with NAC, another potential scavenger, was not significantly different from the control group . These results support the hypothesis that superoxide and hydroxyl radicals contribute to mortality in Gram-negative bacterial sepsis. Am Ind Hyg Assoc J, 1988 Aug, 49(8), 420 - 1 Airborne endotoxin associated with industrial-scale production of protein products in gram-negative bacteria; Palchak RB et al.; Human and animal proteins of therapeutic value can be produced in E . coli, a gram-negative bacteria . Endotoxin, a cellular component, is reported to have clinically significant health effects . Operations--including culturing the microbe, separating solids by centrifugation, and mixing/homogenizing--had associated endotoxin levels ranging from 0.07 ng/m3 to 12.8 ng/m3 . Utilizing a 10-fold safety factor under the threshold where clinically significant changes can be detected, an action level of 30 mg/m3 for large scale operations involving the use of E . coli was established . Operations conducted without engineering controls had maximum airborne endotoxin levels of 1812 ng/m3. Vestn Khir Im I I Grek, 1988 Aug, 141(8), 14 - 7 {Treatment of acute complicated abscess of the lungs}; Grigor'ev EG et al.; Results of examination and treatment of 194 patients with acute complicated abscesses of the lungs were analyzed . The main pathogenic agent of this pathology is gram-negative microorganisms and anaerobic neclostridial flora . Methods of bacterioscopy and gas-liquid chromatography were used for its verification . It was noted that the basis of medical tactics in complicated abscesses of the lungs is the rational antibiotic therapy in combination with parasurgical methods of treatment . Surgical methods are indicated in gangrening abscess, pulmonary hemorrhage, non-effective complex therapy. J Infect Dis, 1988 Aug, 158(2), 312 - 9 Treatment of gram-negative septic shock with human IgG antibody to Escherichia coli J5: a prospective, double-blind, randomized trial; Calandra T et al.; In a randomized, double-blind, multicenter trial we compared the efficacy of a preparation of human IgG antibody to Escherichia coli J5 (J5-IVIG) with that of a standard IgG preparation (IVIG) for the treatment of gram-negative septic shock . At study entry, patients received a single intravenous dose of 200 mg/kg of body weight (maximal dose, 12 g) of either J5-IVIG or IVIG . Of the 100 patients randomized, 71 (30 receiving J5-IVIG and 41 receiving IVIG) had a documented gram-negative infection . Mortality from gram-negative septic shock was 50% (15 of 30) in J5-IVIG recipients and 49% (20 of 41) in IVIG recipients . In addition, treatment with J5-IVIG did not reduce the number of systemic complications of shock and did not delay the occurrence of death due to septic shock . Thus we conclude that J5-IVIG was not superior to IVIG in reducing mortality or in reversing gram-negative septic shock. Jpn J Med Sci Biol, 1988 Aug, 41(4), 123 - 57 Pseudomonas pseudomallei and melioidosis, with special reference to the status in Thailand; Kanai K et al.; Melioidosis is a long-known disease since 1912, but only quite recently we have obtained the knowledges about its actual clinical and epidemiological features . The disease is so unique in having a wide spectrum of disease course and clinical manifestation . The causative agent, P . pseudomallei, is free-living bacterium in the natural environments (soil and surface water) of tropical and subtropical areas . Just like legionnaires' disease, melioidosis is a good example of infectious disease in which pneumonia is produced by inhalation of contaminated soil dusts or water droplets . The infection becomes dormant for years, but with a chance of recrudescence under a variety of insults to the host resistance . The disease, may it be acute or chronic, will be symptomatically confused with malaria, typhoid fever, leptospirosis, septicemia caused by other gram-negative bacteria, tuberculosis and mycotic infections . Isolation of the causative agent from clinical specimens is the only reliable method for diagnosis . Because of the increasing clinical awareness and the development of diagnostic methods, the reported cases of melioidosis have numbered almost one thousand in Thailand during the past 20 years . This country has now the most ample clinical experiences on melioidosis . We have reviewed the history of melioidosis research from bacteriological, immunological, clinical and epidemiological viewpoints, especially including the recent reports in Thailand. J Biochem Biophys Methods, 1988 Aug, 16(4), 301 - 9 A sulfone group-labeled TEM-DNA probe: comparison with a 32P-labeled probe in dot-hybridization; Jouvenot M et al.; A non-radioactive DNA probe for the TEM-type beta-lactamase gene was obtained by using the 'Chemiprobe' system . It was used along with a 32P-labeled TEM probe to screen for TEM beta-lactamase gene in 107 bacterial isolates representing 7 Gram-negative genera and previously classified as TEM-positive or negative . The DNA to be tested was extracted from these bacterial isolates by the Birnboim-Doly method and, after blotting into charged nylon membranes, it was submitted to hybridization with either the TEM 'Chemiprobe' or the 32P-TEM probe . The TEM 'Chemiprobe' could detect as few as 25 pg specific DNA if it was used at a concentration of 5 ng per cm2 of membrane . The results obtained by both probes were concordant in 93.5% of the entire sample . The TEM 'Chemiprobe' was specific since only one false positive was observed . Furthermore, it appeared at least as sensitive as the 32P-labeled TEM probe . As the dot-hybridization with the sulfone-labeled probe was sensitive, simple and easy to perform, it will be useful for large-scale screening in clinical laboratory. Br J Exp Pathol, 1988 Aug, 69(4), 537 - 49 Morphological damage induced by Escherichia coli lipopolysaccharide in cultured hepatocytes: localization and binding properties; Pagani R et al.; Lipopolysaccharides (LPS) from Gram-negative bacteria are considered to be the responsible agents for the induction of endotoxic shock, affecting the liver as a target organ . In this study, the cell morphology and some biochemical properties of 24 h-culture-hepatocyte monolayers treated with Escherichia coli 0111:B4 lipopolysaccharide, were observed . Cell morphology was observed by scanning electron microscopy and immunofluorescence methods . LPS interaction induced an increase in rounded cells with diminished adhesion capacity . As biochemical parameters, albumin synthesis and 2-deoxyglucose uptake were measured . LPS decreased the hexose uptake in a dose-dependent manner . Binding of (14C)LPS to cultured hepatocytes showed that LPS binds to non-specific constituents of the membrane bilayer. J Bacteriol, 1988 Aug, 170(8), 3750 - 1 Penicillin-binding proteins of bdellovibrios; Park JT et al.; We examined the predacious gram-negative bacterium Bdellovibrio bacteriovorous 109J and free-living strains 109J-A1 and 109J-KA1 derived therefrom for penicillin-binding proteins (PBPs) . We compared their PBPs with those of the host bacterium, Escherichia coli, and with those of a facultatively predacious bdellovibrio, B . stolpii UKi2, grown axenically . The multiple PBPs of the 109J strains and of UKi2 differed from each other and from those of E . coli, which suggests that screening for PBPs may be a convenient way to determine to what extent the bdellovibrios may represent a diverse group of organisms . A method for labeling furazlocillin and cefaperizone with iodine-125 is also described. Antibiot Khimioter, 1988 Aug, 33(8), 591 - 4 {The action of combinations of the nonapeptide polymyxin B with antibiotics on gram-negative bacteria}; Bairamashvili DI et al.; Activity of polymyxin B nonapeptide alone and in combination with other antibiotics against clinical strains of Pseudomonas and enteric bacteria was studied . It was shown that nonapeptide was highly active against Pseudomonas and moderately active against enteric bacteria . In combination with rifampicin, fusidic acid or erythromycin the nonapeptide had a potentiating effect on the tested strains. Gene, 1988 Jul 30, 67(2), 301 - 5 Insertion sequence IS2 in the cyanobacterium Chlorogloeopsis fritschii; Machray GC et al.; A cloned DNA fragment, previously demonstrated to encode ribulose bisphosphate carboxylase/oxygenase (RuBisCO) of Chlorogloeopsis fritschii strain CCAP1411/1b, is shown also to include the entire transposable element, IS2, normally a resident in the Escherichia coli genome . Southern-blot hybridisation experiments confirm the presence of IS2 in the C . fritschii genome . This finding adds a new and unrelated species to the known host range of this element and provides evidence of genetic transfer between the Gram-negative E . coli and cyanobacteria . This may also have significance in relation to the nucleotide sequence rearrangements known to occur adjacent to RuBisCO and nif genes in other nitrogen-fixing cyanobacteria. Biochem J, 1988 Jul 15, 253(2), 371 - 6 A novel aspect of the inhibition by arsenicals of binding-protein-dependent galactose transport in gram-negative bacteria; Richarme G; The inhibitory effects of arsenate and arsenite on binding-protein-dependent transport systems are reconsidered . It is shown that arsenate inhibits binding-protein-dependent galactose transport in proteoliposomes energized either by dihydrolipoamide and NAD+ or by a membrane potential (under conditions where ATP metabolism is not implicated); this result is in contradiction with the current interpretation of arsenate inhibition of binding-protein-dependent transport systems (which is based on ATP depletion) and can be explained by reference to the recently discovered ATP inhibition of the binding-protein-dependent galactose transport . In whole cells, the greater inhibition by arsenate of lipoamide-dependent transport than of protonmotive-force-dependent transport may be explained by a modification by arsenate of the pools of several compounds metabolized by 2-oxo-acid dehydrogenases (which have been implicated in binding-protein-dependent transport) . The inhibition of binding-protein-dependent galactose transport by arsenite is probably linked to the inhibition by arsenite of the galactose-stimulated lipoamide dehydrogenase activity implicated in this transport and is reminiscent of the known arsenite inhibition of lipoamide dehydrogenases. Nature, 1988 Jul 14, 334(6178), 173 - 5 Phosphatidylglycerol is involved in protein translocation across Escherichia coli inner membranes; de Vrije T et al.; Newly synthesized proteins to be exported out of the cytoplasm of bacterial cells have to pass across the inner membrane . In Gram-negative bacteria ATP, a membrane potential, the products of the sec genes and leader peptidases (enzymes which cleave the N-terminal signal peptides of the precursor proteins) are required . The mechanism of translocation, however, remains elusive . Important additional roles for membrane lipids have been repeatedly suggested both on theoretical grounds and on the basis of experiments with model systems but no direct evidence had been obtained . We demonstrate here, using mutants of Escherichia coli defective in the synthesis of the major anionic membrane phospholipids, that phosphatidylglycerol is involved in the translocation of newly synthesized outer-membrane proteins across the inner membrane. Drug Intell Clin Pharm, 1988 Jul-Aug, 22(7-8), 577 - 81 Endotracheal tobramycin in gram-negative pneumonitis; Stillwell PC et al.; This report describes the treatment of resistant gram-negative pneumonitis in a compromised host by the combined use of intravenous and endotracheal tobramycin . The endotracheal administration appeared to have an effect on the serum concentration and elimination rate, necessitating a reduction in the amount of drug given intravenously . The only apparent clinical complication of endotracheal drug administration was transient coughing . The addition of endotracheal aminoglycosides to intravenous antibiotics may be useful in pediatric patients with unresponsive (or other difficult-to-treat) pneumonitis caused by resistant microorganisms . The potential contribution of endotracheal aminoglycosides to the serum level and/or disposition profile must be recognized, and therapeutic drug monitoring guided accordingly when this route of administration is used. Appl Environ Microbiol, 1988 Jul, 54(7), 1756 - 60 Efficacy of chemical dosing methods for isolating nontuberculous mycobacteria from water supplies of dialysis centers; Carson LA et al.; Investigations of nontuberculous mycobacterium (NTM) infections associated with various environmental sources have been hampered by the lack of adequate techniques for selective isolation of these organisms from environmental fluids . This study compared chemical dosing techniques for recovery of NTM from water samples collected from 115 randomly selected dialysis centers . Cell suspensions of NTM group II and IV isolates and gram-negative bacteria were exposed to solutions containing sodium hypochlorite (0.2 micrograms/ml of free available chlorine), formaldehyde (1, 0.75, or 0.5%), oxalic acid (1.25%), cetylpyridinium chloride (25 micrograms/ml), or cetyltrimethylammonium bromide (100 micrograms/ml) . Results of standard membrane filtration assays with laboratory test strains and water samples from dialysis centers showed that 5 min of exposure to 1% formaldehyde effectively reduced gram-negative bacterial populations and allowed increased recovery of NTM in environmental fluids containing mixed microbial populations. J Pediatr Orthop, 1988 Jul-Aug, 8(4), 445 - 9 Kingella kingae infection in healthy children; Gamble JG et al.; Kingella kingae is a gram-negative occasional, but normal, inhabitant of the nasopharynx . We present two new cases of this infection that occurred in previously healthy children, and compare and contrast them to other cases reported in the literature . K . kingae osteomyelitis generally has an insidious, subacute onset, whereas septic arthritis has an acute presentation . To date, all strains of K . kingae have been sensitive to penicillin, and no residual damage has been reported following osteomyelitis or septic arthritis, except that residual disk space narrowing did occur after K . kingae discitis. Vet Q, 1988 Jul, 10(3), 211 - 6 Pharmacokinetics of ciprofloxacin in carp, African catfish and rainbow trout; Nouws JF et al.; The plasma disposition of ciprofloxacin was studied in carp, African catfish and trout after intravenous (IV) and intramuscular (IM) administration at a dose rate of 15 mg/kg . Pharmacokinetic analysis of IV data showed that ciprofloxacin was well distributed (distribution volume Vd(area): 3.08-5.59 litre/kg) and exhibited a similar elimination half-life of about 14 h in these 3 fish species . After IM administration to carp and trout a rapid absorption was noticed; the maximum ciprofloxacin plasma concentrations (mean: 3.49 and 2.37 micrograms/ml, respectively), were achieved within 1 h after injection . At the dose level applied, ciprofloxacin has potential therapeutic value for 2-5 days especially against gram-negative bacterial fish pathogens. J Reprod Med, 1988 Jul, 33(7), 657 - 60 Peripartum infections with Capnocytophaga . A case report; Hager H et al.; Capnocytophaga is an increasingly recognized genus of pathogens . Originally associated with periodontal disease, this fastidious, gram-negative organism