Adv Dent Res, 1997 Apr, 11(1), 43 - 9 Phagocyte-bacteria interactions; Keisari Y et al.; Recognition and phagocytosis of micro-organisms in a serum-poor environment represent innate immunity against many extracellular pathogens . As a paradigm for such processes, we discuss the recognition of Klebsiella pneumoniae by alveolar macrophages and monocyte-derived macrophages in the absence of serum . Macrophages recognize and subsequently kill Klebsiella expressing Man-alpha 2/3-Man or Rha-alpha 2/3-Rha sequences in their capsular polysaccharides by two mechanisms: (a) recognition of the capsular structures by macrophage mannose receptors, and (b) opsonization by the lung surfactant protein A (SP-A), which binds to the capsular polysaccharides of Klebsiella and to SP-A receptors on the macrophages . Sp-A may also enhance phagocytosis by increasing the activity of macrophage mannose receptors . We conclude that a specific microbial surface structure may be a target for recognition by macrophages via several mechanisms, as exemplified in the case of Klebsiella capsular polysaccharides . Multiple recognition mechanisms of pathogens by macrophages may be essential to provide innate immunity to reduce the frequency of infections caused by a relatively less virulent bacterium in the immuno-compromised host. Biochim Biophys Acta, 1998 Mar 4, 1396(1), 8 - 14 Cloning and expression of Escherichia coli 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase: identification of the pfs gene product; Cornell KA et al.; The enzyme 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (EC 3.2.2.9) is responsible for cleavage of the glycosidic bond in both 5'-methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH) . Based on amino acid sequence analysis of this enzyme from Klebsiella, we recently speculated that an open reading frame found in E . coli (designated pfs) encoded MTA/SAH nucleosidase . To explore this possibility, we amplified, cloned, and expressed the complete pfs gene from E . coli genomic DNA . The recombinant protein exhibited a molecular weight and Michaelis constants for MTA that are in agreement with those reported for native enzyme . From this biochemical evidence we confirm our original assignment of the pfs gene as encoding MTA/SAH nucleosidase. Microbios, 1997, 91(368-369), 137 - 43 Hydrophobicity and serum sensitivity of Klebsiella pneumoniae treated with sub-MICs of quinolones; Hostacka A; Cell surface hydrophobicity and serum sensitivity of Klebsiella pneumoniae, after treatment with sub-minimum inhibitory concentrations (sub-MICs) of ciprofloxacin, norfloxacin and enoxacin, were studied . The quinolones tested at 1/4, 1/8 and 1/16 of the MICs decreased surface hydrophobicity . The most significant reduction of the bacterial cell surface hydrophobicity was found after treatment with antibiotics at 1/16 of the MICs (to 20.3% for both ciprofloxacin and norfloxacin, and to 23.6% for enoxacin, compared with control values) . The most significant increase in the susceptibility of the bacteria to serum bactericidal activity was seen after 180 min incubation with ciprofloxacin and enoxacin at 1/16 of their MICs . Survival of treated bacteria was 55 +/- 8% 56 +/- 10% as compared with controls without antibiotics. Kaohsiung J Med Sci, 1998 Jan, 14(1), 19 - 24 Ultrasound-guided percutaneous transhepatic drainage of gallbladder followed by cholecystectomy for acute cholecystitis--10 years' experience; Wong SR et al.; Acute cholecystitis is a common disease which may carry the risk of complications, including empyema, perforation, abscess, peritonitis and sepsis . Percutaneous transhepatic drainage of the gallbladder (PTGBD) with antibiotics can provide prompt decompression of gallbladder in acute cholecystitis and interrupt the natural history of the disease effectively . From July 1986 to June 1996, 154 patients with acute cholecystitis were reviewed retrospectively in Kaohsiung Medical College Hospital . The chief symptoms and signs were pain (98.1%), fever (57.1%) and jaundice (37.7%) . WBC count more than 10,000 was noted in 116 (75.3%) patients . Associated diseases included empyema: 42 (27.3%), septic shock: 14 (9.1%), diabetes mellitus: 13 (8.4%), pancreatitis: 10 (6.5%), perforation: 7 (4.5%), liver cirrhosis: 6 (3.9%) and respiratory failure: 1 (0.6%) . All of them underwent ultrasound-guided PTGBD immediately after the diagnosis was established . The symptoms and signs disappeared soon after this procedure . Bacterial culture was found positive in 104 (67.5%) of 154 patients in which Escherichia coli (51.9%) was the most common organism, followed by Klebsiella pneumonia (20.2%) . After acute stage, 138 patients obtained the cholangiography via PTGBD tube . Gallbladder stones were only noted in 56 (40.6%) patients, gallbladder stone concomitant with common bile duct stone in 26 (18.8%), cystic duct obstruction in 25 (18.1%), acalculous cholecystitis in 21 (15.2%), gallbladder perforation in 1 (0.7%), choledochocyst in 1 (0.7%), and cholecystocolonic fistula in 1 (0.7%) . There were 135 patients to undergo surgery after the clinical condition was stable . The operative findings included gallbladder stones only in 88 (65.2%), gallbladder stone concomitant with common bile duct stone in 34 (25.2%), acalculous cholecystitis in 13 (9.6%), choledochocyst in 1 (0.7%), and cholecysto-colonic fistula in 1 (0.7%) . The postoperative complications included wound infection 8 (5.9%), UGI bleeding 3 (2.2%), acute renal failure 1 (0.7%) and acute respiratory failure 1 (0.7%) . The postoperative mortality rate was 0.7% (1/135), which was much lower than those of previous reports, which not undergoing PTGBD initially . It led us to conclude that PTGBD, as an initial preoperative modality to treat acute cholecystitis, is effective in decreasing postoperative morbidity and mortality. Arch Surg, 1998 Mar, 133(3), 242 - 5 Pyogenic liver abscesses in patients with malignant disease: a report of 52 cases treated at a single institution; Yeh TS et al.; BACKGROUND: Prognosis of pyogenic liver abscesses in patients with malignant disease is generally considered poor . The discrepancy between the outcomes of liver abscesses caused by hepatopancreatobiliary malignant disease and those caused by other malignant diseases, however, to our knowledge has never been investigated . OBJECTIVES: To clarify the clinical course of pyogenic liver abscess in patients with different types of cancer, and to compare outcomes in abscesses caused by hepatopancreatobiliary malignant disease and other malignant disease . DESIGN: Retrospective review of case series in our experience from 1980 through 1993 . SETTING: Tertiary care university teaching hospital . PATIENTS: Fifty-two patients with pyogenic liver abscess related to the underlying cancer were divided into 2 groups . Group 1 (n=32) was composed of patients with cancer originating from the hepatic parenchyma, bile duct, and pancreas; group 2 (n=20) was composed of patients with cancer originating from other sites . INTERVENTIONS: Parenteral antibiotics, percutaneous drainage, surgical drainage, or hepatectomy, in combinations, were employed . MAIN OUTCOME MEASURES: Patient characteristics, symptoms, laboratory data, abscess characteristics, microbiological study, management, and outcome of the 2 groups were analyzed . RESULTS: Thirteen patients (41%) in group 1 and 16 patients (80%) in group 2 had undergone prior anticancer treatment . Jaundice was encountered more often in group 1 than in group 2 (29 patients {91%} vs 6 patients {30%}, respectively, P=.001), whereas nausea and vomiting were more frequently seen in group 2 than in group 1 (17 patients {52%} vs 6 patients{31%}, respectively, P=.04) . Leukocytosis, hypoalbuminemia, hyperbilirubinemia, and reversed albumin-globulin ratio were more pronounced in group 1 than in group 2 (P=.001, .02, .003, and .03, respectively) . Abscesses communicating with the intrahepatic biliary tree were more frequently encountered in group 1 than in group 2 (11 patients {34%} vs 2 patients {10%}, respectively, P=.03) . Escherichia coli and Klebsiella pneumoniae predominated in group 1, while the bacteria species in group 2 were more diverse . The hospital mortality rates of group 1 and group 2 were 28% (9 of 32 patients) vs 10% (2 of 20 patients) (P=.04), respectively . Twenty-three patients (72%) of group 1 died of uncontrolled biliary sepsis or progressive cancer or both within 6 months after the diagnosis, while 17 patients (85%) of group 2 survived longer than 1 year without relapse of the abscess and continued with anticancer treatment . CONCLUSIONS: Pyogenic liver abscess could be a presentation of hepatopancreatobiliary malignant disease at the preterminal stage, and carries a grave prognosis . Pyogenic liver abscess in patients with nonhepatopancreatobiliary malignant disease has a better chance of favorable outcome. Plasmid, 1998, 39(2), 123 - 33 Characterization of mutants of the 6'-N-acetyltransferase encoded by the multiresistance transposon Tn1331: effect of Phen171-to-Leu171 and Tyr80-to-Cys80 substitutions; Panaite DM et al.; The Klebsiella pneumoniae plasmid pJHCMW1 harbors a copy of Tn1331, a multiresistance transposon that includes the aac(6')-Ib gene which encodes a 6'-N-aminoglycoside acetyltransferase . This gene was mutagenized using the mutator Escherichia coli XL1-Red . Two plasmids with a single nucleotide mutation in aac(6')-Ib were selected for further analysis: pDP1 and pDP6 . Plasmid pDP1 codes for a mutant enzyme, AAC(6')-IbDP1, that has the Phe171 replaced by a Leu residue . This mutant derivative showed a lower specific activity than the wild-type enzyme when either kanamycin (Km) or its semisynthetic derivative amikacin (Ak) were used as substrates in enzymatic assays performed at 30 degrees C . Furthermore, AAC(6')-IbDP1 showed a change of specificity of substrate when incubated at 42 degrees C . While its acetylating activity for Km was higher at this temperature than at 30 degrees C, it had its ability to utilize Ak as substrate for acetylation considerably reduced . Accordingly, minimal inhibitory concentration assays showed that E . coli(pDP1) was resistant to Ak at 37 degrees C but susceptible at 42 degrees C . The same assays showed that E . coli(pDP1) was highly resistant to Km at either 37 degrees C or 42 degrees C . A high level of resistance to Ak was observed for E . coli(pJHCMW1) which harbors the wild-type AAC(6')-Ib at either 37 or 42 degrees C . Extension of the analyses to other aminoglycosides showed that the enzymatic activity of AAC(6')-IbDP1 as well as the E . coli(pDP1) MICs for netilmicin dropped at 42 degrees C as was the case for Ak . These results could indicate that at 42 degrees C the mutant adopts a conformation that makes it unable to efficiently acetylate aminoglycoside molecules substituted in the C-1amino group of the deoxystreptamine moiety . Plasmid pDP6 encodes the mutant AAC(6')-IbDP6 which has the Tyr80 substituted by a Cys residue . E . coli(pDP6) exhibited reduced MICs for Ak, Km, tobramycin, and netilmicin . Analysis of the acetylating activity of AAC(6')-IbDP6 showed only marginal levels of activity when either Ak, Km, tobramycin, or netilmicin were used as substrates. Alcohol Clin Exp Res, 1998 Feb, 22(1), 157 - 62 Adenoviral-mediated interferon-gamma gene therapy augments pulmonary host defense of ethanol-treated rats; Kolls JK et al.; Alcohol has long been recognized as an immunosuppressive drug and a risk factor for a spectrum of infectious diseases . Among these infections, bacterial pneumonias are most closely correlated with alcohol abuse . One potential mechanism of ethanol-induced immunosuppression is through its ability to suppress alveolar macrophage production of tumor necrosis factor (TNF-alpha) . This defect can be reversed by priming macrophages with interferon-gamma (IFN-gamma) . We hypothesized that macrophage priming in vivo in a model of acute ethanol intoxication could augment pulmonary host defenses . To test this hypothesis, we used adenoviral-mediated gene transfer of the IFN-gamma gene . This strategy resulted in prolonged expression of IFN-gamma in vivo . Moreover, in a model of acute ethanol intoxication, this vector significantly enhanced lipopolysaccharide-induced TNF-alpha responses and lung polymorphonuclear leukocyte recruitment . Furthermore, pulmonary host defenses against Klebsiella pneumoniae were significantly augmented . These enhanced host defenses were not reversed with pretreatment with a polyclonal anti-TNF-alpha antibody, suggesting that IFN-gamma's effect was through a non-TNF-alpha-dependent mechanism . These data demonstrate that ethanol-induced suppression of pulmonary host defenses can be reversed with IFN-gamma gene therapy. J Med Microbiol, 1998 Mar, 47(3), 201 - 9 Long-term investigation of the clonal dissemination of Klebsiella pneumoniae isolates producing extended-spectrum beta-lactamases in a university hospital; Branger C et al.; Seventy isolates of Klebsiella pneumoniae with extended-spectrum beta-lactamases (ESBLs) were compared . These were isolated from 51 patients on 10 separate wards in one hospital over an 18-month period between 1992 and 1994 . Antibiograms were determined and the isolates were typed by pulsed-field gel electrophoresis of their DNA digestion with XbaI . The isolates were compared to three genotypically different epidemic strains responsible for previous outbreaks at the hospital between 1988 and 1991 . Isolates from 84% of the present patients had closely related XbaI patterns, and most (74%) produced an ESBL with an iso-electric point (pI) of 7.0 . A similar pattern was found for one of the previous epidemic strains, but it produced an ESBL with a pI of 7.8; isolates with this latter enzyme variant were found only in six of the present patients . The two other previous epidemic strains had ESBLs with a pI of 6.3 and organisms related to them were found in one and two of the present patients, respectively. Epidemiol Mikrobiol Imunol, 1998 Feb, 47(1), 12 - 4 {Effectiveness of cefpirome on Klebsiella pneumoniae producing beta-lactamases with an extended spectrum of activity}; Kolar M; In the submitted paper the author investigated the effectiveness of cefpirome on Klebsiella pneumoniae strains producing beta-lactamases with an extended spectrum (ESBL) . From a total of 1348 examined strains of Klebsiella pneumoniae ESBL production was confirmed in 59 strains (4.4%) . Resistance of ESBL-positive strains of Klebsiella pneumoniae to ceftazidime was 83.1%, to cefotaxime 93.2%, while resistance to cefpirome was recorded only in 45.8%. J Antimicrob Chemother, 1998 Jan, 41(1), 123 - 5 PCR single strand conformational polymorphism can be used to detect the gene encoding SHV-7 extended-spectrum beta-lactamase and to identify different SHV genes within the same strain; M'Zali FH et al.; The PCR single strand conformational polymorphism (PCR-SSCP) technique described to identify mutants of the SHV beta-lactamases was extended to identify an SHV-7 type beta-lactamase . This was found in a strain of Klebsiella pneumoniae, the first recorded isolate in the UK to produce this type of enzyme . We also demonstrate that PCR-SSCP can be used to identify more than one SHV beta-lactamase gene in a single strain. Kansenshogaku Zasshi, 1998 Jan, 72(1), 75 - 82 Effects of cytokines and minocycline on subacute lung injuries induced by repeated injection of lipopolysaccharide; Yamaki K et al.; Pathological changes were seen in the lungs of ddY mice one week after repeated intraperitoneal injections of lipopolysaccharide (LPS) of Klebsiella pneumoniae . The infiltration of polymorphonuclear cells (PMN), mainly neutrophils, and lymphocytes into the alveolar septum, the infiltration of PMN into perivascular areas and microthrombi were recognized in this murine model . The blood levels of TNF alpha and IL-1 alpha did not rise at this time, suggesting that the most important cytokine promoting inflammation one week after LPS stimulation was neither TNF alpha nor IL-1 alpha . In the lungs of mice administered minocycline together with LPS, lymphocyte infiltration of alveoli and perivascular areas as well as microthrombi were suppressed . The blood levels of TNF alpha, IL-1 alpha, IL-4 and IL-6 were elevated in these groups, suggesting the suppression of pathological changes to be associated with the anti-inflammatory effect of IL-6 and/or persistent elevation of TNF alpha and/or IL-1 alpha levels . In conclusion, subacute pathological changes in the lung were induced by repeated intraperitoneal injections of LPS to mice . These pathological changes were suppressed by minocycline, suggesting the anti-inflammatory effects of this antibiotic to be the result of stimulating certain cytokines. Clin Infect Dis, 1998 Feb, 26(2), 468 - 74 Klebsiella endocarditis: report of two cases and review; Anderson MJ et al.; The rarity of endocarditis due to Klebsiella species limits its recognition and awareness of its often malignant course . We describe two recent cases of Klebsiella pneumoniae endocarditis and review the clinical context and outcomes of 48 other cases reported in the literature . At our hospital, endocarditis complicated only one of 86 consecutive episodes of bacteremia due to Klebsiella species . In 22 series of endocarditis that we reviewed, Klebsiella species caused < or =1.2% of cases of native valve endocarditis and up to 4.1% of cases of prosthetic valve endocarditis . Valves were replaced in 44% of these cases, and the mortality rate was 49% in cases for which outcome was specified . Valve replacement may be associated with improved survival . We conclude that Klebsiella species are a rare but ominous cause of complicated bacterial endocarditis that requires careful evaluation during the entire course of therapy. Chem Pharm Bull (Tokyo), 1998 Feb, 46(2), 319 - 23 Some properties and the inclusion behavior of three positional isomers of 6(1),6n-di-O-alpha-D-glucosyl-cyclomaltoheptaoses (beta-cyclodextrins); Okada Y et al.; Three positional isomers of 6(1),6n-di-O-alpha-D-glucosyl-cyclomaltoheptaose {1,n-(G)2-beta CDs; n = 2-4} which existed in the digests with glucoamylase of the products from cyclomaltoheptaose (beta-cyclodextrin, beta CD) and maltose with Klebsiella pneumoniae pullulanase, were purified by HPLC . The solubilities of two isomers of those doubly branched beta CDs, 1,2- and 1,3-(G)2-beta CDs, in water were much higher than those of parent non-branched beta CD and mono-branched beta CD, 6-O-alpha-D-glucosyl-beta CD (G-beta CD), while the solubility of another isomer, 1,4-(G)2-beta CD, was significantly lower than these two isomers, though it was higher than that of beta CD . On the other hand, the solubilities of 1,2- and 1,3-isomers in 10, 30, and 50% (v/v) aqueous methanol at 25 degrees C were independent of methanol concentrations and their solubilities were the same as those in water at 25 degrees C . However, that of 1,4-isomer increased with increasing methanol concentrations . The hemolytic activities of 1,n-(G)2-beta CDs on human erythrocytes in isotonic solution were lower than those of G-beta CD and beta CD, and became weaker in the order of 1,4- > 1,2- > 1,3-isomers . The complex-forming abilities of 1,n-(G)2-beta CDs for digitoxin, digoxin, fluorometholone, flurbiprofen, hydrocortisone acetate, and norfloxacin were about the same as those of beta CD and G-beta CD, whereas reserpine was more difficult to include within 1,n-(G)2-beta CDs than beta CD and G-beta CD . Nevertheless, the solubilities of those guest compounds were much more enhanced by 1,n-(G)2-beta CDs and G-beta CD than by beta CD. Methods Find Exp Clin Pharmacol, 1997 Nov, 19(9), 579 - 83 A low dose immunorestorative effect of aporphinoid alkaloid oxoglaucine on experimentally immunosuppressed and infected mice; Ivanovska N et al.; The immunoregulatory activity of aporphinoid alkaloid oxoglaucine was studied in delayed-type hypersensitivity (DTH) reaction and Klebsiella pneumoniae infection in normal and immunosuppressed mice . The alkaloid enhanced DTH reaction when applied during the developing phase and abolished the inhibitory action of cyclophosphamide . Oxoglaucine possessed a restorative effect in K . pneumoniae infection in immunosuppressed mice in combination with indomethacin, prednisolone or artemisinin, as evaluated by the number of survivors and mean survival time . These data suggest that oxoglaucine affects selected lymphocyte clones responsible for antiinfectious host resistance . Its use as a second agent in combination with another immunosuppressant might enable reduction in the dosage or time of application. Biotechnol Prog, 1998 Jan-Feb, 14(1), 116 - 25 Metabolic engineering of propanediol pathways; Cameron DC et al.; Microbial fermentation is an important technology for the conversion of renewable resources to chemicals . In this paper, we describe the application of metabolic engineering for the development of two new fermentation processes: the microbial conversion of sugars to 1,3-propanediol (1,3-PD) and 1,2-propanediol (1,2-PD) . A variety of naturally occurring organisms ferment glycerol to 1,3-PD, but no natural organisms ferment sugars directly to 1,3-PD . We first describe the fed-batch fermentation of glycerol to 1,3-PD by Klebsiella pneumoniae . We then present various approaches for the conversion of sugars to 1,3-PD, including mixed-culture fermentation, cofermentation of glycerol and glucose, and metabolic engineering of a "sugars to 1,3-PD" pathway in a single organism . Initial results are reported for the expression of genes from the K . pneumoniae 1,3-PD pathway in Saccharomyces cerevisiae . The best naturally occurring organism for the fermentation of sugars to 1,2-PD is Thermoanaerobacterium thermosaccharolyticum . We describe the fermentation of several different sugars to 1,2-PD by this organism in batch and continuous culture . We report that Escherichia coli strains engineered to express either aldose reductase or glycerol dehydrogenase convert glucose to (R)-1,2-PD . We then analyze the ultimate potential of fermentation processes for the production of propanediols . Linear optimization studies indicate that, under aerobic conditions, propanediol yields that approach the theoretical maximum are possible and CO2 is the primary coproduct . Without the need to produce acetate, final product titers in the range of 100 g/L should be possible; the high titers and low coproduct levels should make product recovery and purification straightforward . The examples given in this paper illustrate the importance of metabolic engineering for fermentation process development in general. J Bacteriol, 1998 Mar, 180(5), 1311 - 22 NasFED proteins mediate assimilatory nitrate and nitrite transport in Klebsiella oxytoca (pneumoniae) M5al; Wu Q et al.; Klebsiella oxytoca can use nitrate and nitrite as sole nitrogen sources . The enzymes required for nitrate and nitrite assimilation are encoded by the nasFEDCBA operon . We report here the complete nasFED sequence . Sequence comparisons indicate that the nasFED genes encode components of a conventional periplasmic binding protein-dependent transport system consisting of a periplasmic binding protein (NasF), a homodimeric intrinsic membrane protein (NasE), and a homodimeric ATP-binding cassette (ABC) protein (NasD) . The NasF protein and the related NrtA and CmpA proteins of cyanobacteria contain leader (signal) sequences with the double-arginine motif that is hypothesized to direct prefolded proteins to an alternate protein export pathway . The NasE protein and the related NrtB and CmpB proteins of cyanobacteria contain unusual variants of the EAA loop sequence that defines membrane-intrinsic proteins of ABC transporters . To characterize nitrate and nitrite transport, we constructed in-frame nonpolar deletions of the chromosomal nasFED genes . Growth tests coupled with nitrate and nitrite uptake assays revealed that the nasFED genes are essential for nitrate transport and participate in nitrite transport as well . Interestingly, the delta nasF strain exhibited leaky phenotypes, particularly at elevated nitrate concentrations, suggesting that the NasED proteins are not fully dependent on the NasF protein. Eur Respir J, 1997 Dec, 10(12), 2902 - 3 A case of superior vena cava syndrome caused by Klebsiella pneumoniae; Kim JY et al.; A 27 yr old man presented with productive cough, fever and manifestations of superior vena cava syndrome . He was an alcoholic but had been in good health until 3 days prior to admission . The physical examination, the chest radiograph and the results of the sputum culture were compatible with Klebsiella pneumoniae pneumonia of the right upper lobe . The superior vena cava scintigram using technetium-99m showed near total occlusion of the superior vena cava, while sputum cytology, chest computed tomography, and bronchoscopy were all negative for malignant aetiology . Antibiotic therapy brought about slow resolution of the pneumonia and also of the superior vena caval obstruction . The follow-up scintigram showed normalized venous flow of the superior vena cava . To our knowledge, this is the first case of superior vena cava syndrome developed in probable association with Klebsiella pneumoniae pneumonia. Microbiology, 1998 Feb, 144 ( Pt 2), 555 - 9 High affinity binding of albicidin phytotoxins by the AlbA protein from Klebsiella oxytoca; Zhang L et al.; Albicidins are a family of phytotoxins and antibiotics which play an important role in the pathogenesis of sugarcane leaf scald disease . The albA gene from Klebsiella oxytoca encodes a protein which inactivates albicidin by heat-reversible binding . Albicidin ligand binding to a recombinant AlbA protein, purified by means of a glutathione S-transferase gene fusion system, is an almost instant and saturable reaction . Kinetic and stoichiometric analysis of the binding reaction indicated the presence of a single high affinity binding site with a dissociation constant of 6.4 x 10(-8) M . The AlbA-albicidin complex is stable from 4 to 40 degrees C, from pH 5 to 9 and in high salt solutions . Treatment with protein denaturants released all bound albicidin . These properties indicate that AlbA may be a useful affinity matrix for selective purification of albicidin antibiotics . AlbA does not bind to p-nitrophenyl butyrate or alpha-naphthyl butyrate, the substrates of the albicidin detoxification enzyme AlbD from Pantoea dispersa . The potential exists to pyramid genes for different mechanisms in transgenic plants to protect plastid DNA replication from inhibition by albicidins. Microbiology, 1998 Feb, 144 ( Pt 2), 343 - 52 Escherichia coli CoIV plasmid pRK100: genetic organization, stability and conjugal transfer; Ambrozic J et al.; Uropathogenic Escherichia coli strains express chromosomal and plasmid-encoded virulence-associated factors such as specific adhesins, toxins and iron-uptake systems . A CoIV plasmid (pRK100) of a uropathogenic strain and its host KS533 were studied . The host strain encodes the K1 capsule, and P and S fimbriae, but neither haemolysin nor the cytotoxic-necrotic factor CNF1, indicating that this strain does not harbour a larger pathogenicity island . A restriction map of pRK100 was constructed on the basis of hybridization experiments and nucleotide sequencing . pRK100 harbours CoIV, the conserved replication region RepFIB, the aerobactin-uptake system, a RepFIC replicon and additionally Colla as well as transposon Tn5431 . The location of the RepFIC replicon was similar to that in plasmid F . CoIV plasmids and F thus share a region spanning more than half the length of plasmid F . Even though their replication and transfer regions are homologous, CoIV plasmids are found only in E . coli strains . Among the four other species tested, conjugal transfer of pRK100 was demonstrated, with low frequency, only to Klebsiella pneumoniae, suggesting that a natural barrier effectively bars transfer . In vitro stability of the plasmid with integration into the chromosome to ensure maintenance in the presence of an incompatible plasmid was demonstrated. J Clin Apheresis, 1997, 12(4), 200 - 1 Plasmapheresis in the treatment of inadvertent intravenous infusion of an enteral feeding solution; Ong BC et al.; This 50-year-old man was on enteral full-strength Ensure through the jejunostomy tube . The enteral feed was accidentally infused through the peripheral intravenous line . He went into a septic state with hypotension, hypoxemia, and increased pulmonary shunting requiring ventilatory and inotropic support . We instituted two cycles of plasmapheresis in an attempt to remove any foreign antigen and toxins present . This resulted in improved oxygenation, rapid recovery, and discharge from intensive care . His blood culture grew Klebsiella species that was consistent with the organism grown from the culture of the Ensure feeds. Am J Perinatol, 1998 Jan, 15(1), 47 - 51 Piperacillin/tazobactam in the treatment of Klebsiella pneumoniae infections in neonates; Pillay T et al.; Nosocomial Klebsiella pneumoniae infection is associated with a high mortality in neonates and antimicrobial therapy of these infections has been complicated by the emergence of multiresistant strains . These organisms remain susceptible to only a few antimicrobial agents, and some of these are not recommended for use in children . In this study the antimicrobial agents used in the treatment of 33 neonates with Klebsiella pneumoniae (K . pneumonia) infection in our tertiary neonatal unit, during an outbreak were: piperacillin/tazobactam (13), imipenem/cilastatin (17), cefotaxime (2), and ciprofloxacin (1) . Extended-spectrum beta-lactamase production was detected in K . pneumoniae isolates from 18 of 33 (54.5%) neonates . All-cause mortality was 13 of 33 (39.4%) and there was no significant difference in mortality between neonates treated with imipenem/cilastatin (6 of 17 or 35.3%) and neonates treated with piperacillin/tazobactam (6 of 13 or 46.2%) . The duration of antimicrobial therapy and total hospital stay was similar between neonates who received imipenem/cilastatin and those that received piperacillin/tazobactam . This report suggests that piperacillin/tazobactam may be a useful antimicrobial agent in neonatal infections caused by beta-lactamase-producing organisms. Am J Gastroenterol, 1998 Feb, 93(2), 253 - 5 Cholangiocarcinoma presenting as pyogenic liver abscess: is its outcome influenced by concomitant hepatolithiasis? Jan YY, Yeh TS, Chen MF. OBJECTIVE: The etiology of pyogenic liver abscess is changing . Malignant biliary obstruction has emerged as one of the most important causes . We explored the clinical course of pyogenic liver abscess caused by cholangiocarcinoma . METHODS: The medical records of 19 patients with cholangiocarcinoma presenting as pyogenic liver abscess were reviewed . Of them, 57.8% (11 of 19) had concomitant hepatolithiasis . Escherichia coli and Klebsiella pneumoniae were the most common pathogens isolated from aspirates of the abscesses . Eight patients received percutaneous drainage, whereas 11 patients initially underwent surgical drainage because of the presence of ascites or coagulopathy or lack of awareness of the underlying cholangiocarcinoma . RESULTS: Overall, the hospital mortality rate was 36.8% (seven of 19) . Patients with hepatolithiasis had a hospital mortality rate of 54.5% (six of 11), compared with 12.5% (one of eight) in those without (p < 0.01) . Notably, 84.2% (16 of 19) of the patients died within 6 months after the diagnosis was made . CONCLUSIONS: Cholangiocarcinoma presenting as liver abscess has a dismal prognosis . Concomitant hepatolithiasis worsened the infectious process and adversely affected the survival. J Antimicrob Chemother, 1997 Dec, 40(6), 789 - 95 Characterization and amino acid sequence of the OXY-2 group beta-lactamase of pI 5.7 isolated from aztreonam-resistant Klebsiella oxytoca strain HB60; Farzaneh S et al.; Klebsiella oxytoca strain HB60 is highly resistant to cefoperazone and aztreonam (MICs = 128 mg/L) . It produces a chromosomally encoded beta-lactamase of pI 5.7 which was highly efficient against penicillins, first-generation cephalosporins and cefoperazone, a non-oxyimino third-generation cephalosporin . Aztreonam and oxyimino broad-spectrum cephalosporins were less good substrates . The beta-lactamase activity was susceptible to inhibition by clavulanic acid (IC50 = 1 microM) . The enzyme purified to homogeneity had a specific activity towards benzylpenicillin of 3670 U/mg . The 263 amino acid residues of the protein were sequenced by Edman degradation of proteolytic peptides . The beta-lactamase was shown to belong to the OXY-2 group as it had only one amino acid substitution (Asn for Asp at ABL position 197) compared with the beta-lactamase (pI 5.2) from the aztreonam-susceptible K . oxytoca strain SL911 and two substitutions (Ala223 for Val and Asp255 for Asn) compared with the beta-lactamase (pI 6.4) from the aztreonam-resistant K . oxytoca strain D488 . These three OXY-2-group enzymes behave in the same way towards beta-lactam antibiotics . The variability in the resistance of these K . oxytoca strains would thus seem to be due to variation in the level of production of the beta-lactamases rather than to structural alteration of the enzymes. J Clin Microbiol, 1998 Jan, 36(1), 266 - 8 Sporadic emergence of Klebsiella pneumoniae strains resistant to cefepime and cefpirome in Greek hospitals; Tzouvelekis LS et al.; The sporadic emergence of Klebsiella pneumoniae strains resistant to cefepime and cefpirome was observed in Greek hospitals during 1996 . Examination of six epidemiologically distinct strains and clones selected in vitro provided indications that resistance is due to the cooperation of decreased outer membrane permeability and hydrolysis of the cephalosporins by SHV-5 beta-lactamase, which was produced in large amounts. Klin Lab Diagn, 1997 Aug, (8), 38 - 41 {Methodological aspects of Klebsiella pneumoniae serotyping and its use for characterization of clinical isolates}; Morozova OT et al.; The method for serological typing of Klebsiella pneumoniae making use of Russian commercial K-sera manufactured by the Ilya Metchnikoff firm has been used to characterize 85 strains isolated from newborns at an obstetrical hospital and department of newborn diseases and from children with acute enteric infections hospitalized at the hospital for infectious diseases . The authors emphasize that their methods of serotyping are to be accurately performed, specifically, the selection of capsular forms and identification of serovars in strains which can be agglutinated by several sera . Serovars were identified by the proposed serotyping method in 89.4% of the studied strains . A wide spectrum of K-serovars typical of this or that hospital has been defined for each institution . K . pneumoniae K2 predominated in the obstetrical hospital, K . pneumoniae K24 and K25 prevailed in department for newborn diseases, and the K14 variant in the infectious diseases hospital. J Bacteriol, 1998 Feb, 180(3), 578 - 85 Two roles for the DNA recognition site of the Klebsiella aerogenes nitrogen assimilation control protein; Pomposiello PJ et al.; The nitrogen assimilation control protein (NAC) binds to a site within the promoter region of the histidine utilization operon (hutUH) of Klebsiella aerogenes, and NAC bound at this site activates transcription of hutUH . This NAC-binding site was characterized by a combination of random and directed DNA mutagenesis . Mutations that abolished or diminished in vivo transcriptional activation by NAC were found to lie within a 15-bp region contained within the 26-bp region protected by NAC from DNase I digestion . This 15-bp core has the palindromic ends ATA and TAT, and it matches the consensus for LysR family transcriptional regulators . Protein-binding experiments showed that transcriptional activation in vivo decreased with decreasing binding in vitro . In contrast to the NAC-binding site from hutUH, the NAC-binding site from the gdhA promoter failed to activate transcription from a semisynthetic promoter, and this failure was not due to weak binding or greatly distorted protein-DNA structure . Mutations in the promoter-proximal half-site of the NAC-binding site from gdhA allowed this site to activate transcription . Similar studies using the NAC-binding site from hut showed that two mutations in the promoter proximal half-site increased binding but abolished transcriptional activation . Interestingly, for symmetric mutations in the promoter-distal half-site, loss of transcriptional activation was always correlated with a decrease in binding . We conclude from these observations that if the binding in vitro reflects the binding in vivo, then binding of NAC to DNA is not sufficient for transcriptional activation and that the NAC-binding site can be functionally divided in two half-sites, with related but different functions. Arch Pathol Lab Med, 1997 Dec, 121(12), 1287 - 91 Epstein-Barr virus-associated primary central nervous system lymphoma in a child with the acquired immunodeficiency syndrome . A case report and review of the literature; Rodriguez MM et al.; A 34-month-old black boy who had contracted acquired immunodeficiency syndrome from his mother presented with fever, vomiting, and cough . He was cachectic, hypertonic, and developmentally delayed . A brain computed tomography scan revealed masses in the left frontal horn, subependymal, and periventricular regions; secondary edema; and hydrocephalus . The differential diagnosis was cerebral lymphoma versus toxoplasmosis . The patient had disseminated Mycobacterium avium-intracellulare infection, lymphoid interstitial pneumonitis, as well as Pseudomonas and Klebsiella pneumonia . He died of respiratory insufficiency 53 days after admission . The autopsy confirmed a primary cerebral B-cell lymphoma, large cell type, which was positive for Epstein-Barr virus, latent phase, by in situ hybridization . Primary central nervous system lymphomas are rare in children, in contrast to adults . To our knowledge, only five well-documented cases of primary cerebral lymphomas in infants and children with acquired immunodeficiency syndrome have been reported previously . The current study shows that these childhood lymphomas are associated with and presumably caused by Epstein-Barr virus and thus have a pathogenesis similar to that of primary central nervous system lymphomas in adults. Antimicrob Agents Chemother, 1998 Jan, 42(1), 108 - 13 A novel extended-spectrum TEM-type beta-lactamase (TEM-52) associated with decreased susceptibility to moxalactam in Klebsiella pneumoniae; Poyart C et al.; Klebsiella pneumoniae NEM865 was isolated from the culture of a stool sample from a patient previously treated with ceftazidime (CAZ) . Analysis of this strain by the disk diffusion test revealed synergies between amoxicillin-clavulanate (AMX-CA) and CAZ, AMX-CA and cefotaxime (CTX), AMX-CA and aztreonam (ATM), and more surprisingly, AMX-CA and moxalactam (MOX) . Clavulanic acid (CA) decreased the MICs of CAZ, CTX, and MOX, which suggested that NEM865 produced a novel extended-spectrum beta-lactamase . Genetic, restriction endonuclease, and Southern blot analyses revealed that the resistance phenotype was due to the presence in NEM865 of a 13.5-kb mobilizable plasmid, designated pNEC865, harboring a Tn3-like element . Sequence analysis revealed that the blaT gene of pNEC865 differed from blaTEM-1 by three mutations leading to the following amino acid substitutions: Glu104-->Lys, Met182-->Thr, and Gly238-->Ser (Ambler numbering) . The association of these three mutations has thus far never been described, and the blaT gene carried by pNEC865 was therefore designated blaTEM-52 . The enzymatic parameters of TEM-52 and TEM-3 were found to be very similar except for those for MOX, for which the affinity of TEM-52 (Ki, 0.16 microM) was 10-fold higher than that of TEM-3 (Ki, 1.9 microM) . Allelic replacement analysis revealed that the combination of Lys104, Thr182, and Ser238 was responsible for the increase in the MICs of MOX for the TEM-52 producers. Antimicrob Agents Chemother, 1998 Jan, 42(1), 53 - 8 Epidemiology and successful control of a large outbreak due to Klebsiella pneumoniae producing extended-spectrum beta-lactamases; Pena C et al.; An outbreak due to extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-KP) was detected from May 1993 to June 1995 . A total of 145 patients, particularly patients in intensive care units (ICUs) (107 patients {72%}), were colonized or infected . Infection developed in 92 (63%) patients, and primary bacteremia caused by ESBL-KP was the most frequent infection (40 of 92 patients {43%}) . A single clone of ESBL-KP was identified by pulsed-field gel electrophoresis analysis throughout the whole period, and no molecular epidemiological relationship could be found between the epidemic strain and non-ESBL-KP isolates . To determine risk factors for ESBL-KP infection weekly rectal swabs were obtained in three serial incidence surveys (470 patients); the probabilities of carriage of ESBL-KP in the digestive tract were 33% (October and November 1993), 40% (May and June 1994), and 0% (October and November 1995) at 10 days of ICU admission . A logistic regression model identified prior carriage of ESBL-KP in the digestive tract (odds ratio, 3.4; 95% confidence interval 1.1 to 10.4) as an independent variable associated with ESBL-KP infection . A statistically significant correlation was observed between the restricted use of oxyimino-beta-lactams (189 defined daily doses {DDD}/ 1,000 patient-days to 24 DDD/1,000 patient-days) and the trends of ESBL-KP infection (r = 0.7; P = 0.03). Food Chem Toxicol, 1997 Dec, 35(12), 1151 - 7 The effect of vitamin C on N-acetyltransferase activity in Klebsiella pneumoniae; Hsieh SE et al.; This study was designed to assess the effect of vitamin C on arylamine N-acetyltransferase (NAT) activity in Klebsiella pneumoniae by using HPLC to measure the acetylation of 2-aminofluorene (2-AF) with and without vitamin C . Two assay systems were performed, one with intact bacterial cell suspensions, the other with S-9 fractions (9000g supernatant) . It was found that vitamin C promoted NAT activity in K . pneumoniae in a dose-dependent manner in both systems . 4 and 8 mM vitamin C were selected for further studies in S-9 fractions and intact cell systems, respectively . Through a 4-hr time course study, vitamin C promoted the N-acetylation of 2-AF in both assay systems, but, the longer the reaction time lasted, the lower the promotion rate . In the kinetic studies, vitamin C increased the value of Km from 0.42+/-0.03 mM to 2.43+/-0.87 mM in S-9 fraction assays and from 0.54+/-0.03 mM to 0.85+/-0.18 mM in intact cell assays . Vitamin C also increased the apparent Vmax values from 3.5 +/-0.08 to 39.66+/-9.81 nmol/min/mg protein in S-9 fraction assays, and from 1.28+/-0.06 to 4.88+/-0.87 nmol/min/10(10) CFU in intact cell assays, for acetylation of 2-AF . In the presence of vitamin C, the NAT activity was increased from 0.58+/-0.06 to 1.34+/-0.02 nmol/min/mg protein in S-9 fractions, and from 0.18+/-0.02 to 0.40+/-0.02 nmol/min/10(10) CFU in intact cells, for acetylation of 2-AF . From the present study, it is concluded that vitamin C does promote the N-acetylation of 2-AF in K . pneumoniae . This is a first report suggesting that oral vitamin C may be involved in modifying the mutagenicity/carcinogenicity of ingested arylamines through enhancing the NAT activity of human enteric bacteria . This interaction should be pursued in future in vivo studies. Am J Gastroenterol, 1998 Jan, 93(1), 118 - 9 A new variant of food poisoning: enteroinvasive Klebsiella pneumoniae and Escherichia coli sepsis from a contaminated hamburger; Sabota JM et al.; For the first time, we report Klebsiella pneumoniae as an enteroinvasive food-borne pathogen transmitted from a hamburger . A 28-year-old previously healthy African-American male ingested a portion of a hamburger from a fast food chain . Symptoms of gastroenteritis rapidly deteriorated to multiorgan failure . Blood and hamburger cultures grew Escherichia coli and Klebsiella pneumoniae . Since Klebsiella had not previously been reported as enteroinvasive, the isolates were compared . Full biochemical profiles, antimicrobial sensitivity, plasmid profile, and toxin assay by DNA hybridization probe were completely concordant . The patient survived the episode of food-borne sepsis . Deliberate or inadvertent employee contamination of food products with feces may be a potential source of life-threatening food-borne illness. Zentralbl Bakteriol, 1997 Nov, 286(4), 449 - 55 Transfer of antibiotic resistance and production of extended-spectrum beta-lactamase (ESBL) in nosocomial Klebsiella pneumoniae strains from two hospitals in Czech Republic; Blahova J et al.; Six Klebsiella pneumoniae strains were collected from two hospitals in Ostrava, Czech republic . Four strains (Nos . 209, 217, 218, 222) were isolated from sputa of critically ill patients from Municipal Hospital Vitkovice-Ostrava . They were resistant to cephalothin, cefotaxime, and ceftazidime (MIC > 100 mg x l-1) . Strain No . 218 was intermediately resistant also to ofloxacin and aztreonam (MIC = 12.5 mg x l-1), strain No . 222 was resistant to aztreonam (MIC = 50 mg x l-1) . Determinants of resistance to cephalothin, cefotaxime, aztreonam and ceftazidime were transferred to recipient strains of P . mirabilis P-38 rif+ and E . coli K-12 No . 3110 rif+ by all four strains . Synergy between clavulanate-cefotaxime, clavulanate-ceftazidime and clavulanate-aztreonam indicated production of ESBLs by these strains . Two strains, No . 214 and 224, from patients of the ICU in the University Hospital were resistant to cephalothin, cefotaxime and ceftazidime (MIC > 100 mg x l-1) . Strain No . 214 was intermediately resistant to aztreonam and ofloxacin (MIC = 12.5 mg x l-1) and strain No . 224 was highly resistant to aztreonam (MIC = 50 mg x l-1) . Synergy between clavulanate and cefotaxime as well as between clavulanate and aztreonam, but not between clavulanate and ceftazidime corresponds with non-transferable ceftazidime resistance in strains No . 214 and 224 and indicates different types of ESBL in strains from each of two hospitals. J Biol Chem, 1997 Dec 19, 272(51), 32034 - 41 Characterization, sequencing, and expression of the genes encoding a reactivating factor for glycerol-inactivated adenosylcobalamin-dependent diol dehydratase; Mori K et al.; Diol dehydratase undergoes suicide inactivation by glycerol during catalysis involving irreversible cleavage of the Co-C bond of adenosylcobalamin . In permeabilized Klebsiella oxytoca and Klebsiella pneumoniae cells, the glycerol-inactivated holoenzyme or the enzyme-cyanocobalamin complex is rapidly activated by the exchange of the inactivated coenzyme or cyanocobalamin for free adenosylcobalamin in the presence of ATP and Mg2+ (Honda, S., Toraya, T., and Fukui, S . (1980) J . Bacteriol . 143, 1458-1465; Ushio, K., Honda, S., Toraya, T., and Fukui, S . (1982) J . Nutr . Sci . Vitaminol . 28, 225-236) . Permeabilized Escherichia coli cells co-expressing the diol dehydratase genes with two open reading frames in the 3'-flanking region were capable of reactivating glycerol-inactivated diol dehydratase as well as activating the enzyme-cyanocobalamin complex in situ in the presence of free adenosylcobalamin, ATP, and Mg2+ . These open reading frames, designated as ddrA and ddrB genes, were identified as the genes of a putative reactivating factor for inactivated diol dehydratase . The genes encoded polypeptides consisting of 610 and 125 amino acid residues with predicted molecular weights of 64,266 and 13,620, respectively . Co-expression of the open reading frame in the 5'-flanking region was stimulatory but not obligatory for conferring the reactivating activity upon E . coli . Thus, the product of this gene was considered not an essential component of the reactivating factor. FEMS Microbiol Lett, 1997 Dec 15, 157(2), 313 - 8 NifL of Klebsiella pneumoniae carries an N-terminally bound FAD cofactor, which is not directly required for the inhibitory function of NifL; Schmitz RA; In Klebsiella pneumoniae NifL antagonizes the action of the transcriptional activator NifA in the presence of molecular oxygen or combined nitrogen . To determine what cofactors might be involved in the oxygen sensing mechanism, we purified and analyzed fusion proteins made between the Escherichia coli maltose binding protein, MalE, and NifL . NifL synthesized and purified under strictly anaerobic conditions did not contain significant amounts of iron or acid-labile sulfur indicating the absence of an oxygen sensing iron-sulfur cluster . However, NifL protein purified in its inhibitory form contained 0.3 +/- 0.01 mol FAD and less than 0.01 mol FMN per mol NifL suggesting the presence of FAD as a cofactor . Characterization of NifL synthesized in the absence of oxygen and combined nitrogen showed that the non-inhibitory form of NifL also contained FAD (0.54 mol FAD per mol NifL) . Using fusions between MalE and different portions of NifL we localized the binding site of FAD to the N-terminal domain of NifL . These results and our previous observation that the C-terminal domain of NifL is sufficient to inhibit NifA activity indicate that the N-terminally bound FAD is not directly required for the inhibitory activity of NifL . This observation is supported by the finding that purified apoprotein of NifL was still able to inhibit transcriptional activation by NifA in vitro. Scand J Infect Dis, 1997, 29(5), 491 - 3 Dietary fish oil supplementation increases survival in mice following Klebsiella pneumoniae infection; Bjornsson S et al.; The effect of dietary fish-oil supplementation on survival of NMRI mice after Klebsiella pneumoniae infection was investigated . 30 mice in each group were fed a fish-oil enriched diet, olive-oil enriched diet or standard chow diet . After 6 weeks, the mice were injected intramuscularly with Klebsiella pneumoniae . After 120 h the survival of the mice fed fish-oil enriched diet was 40%, while the survival for mice fed standard or olive-oil enriched diets was 20% and 25%, respectively . The survival curve over 120 h was significantly improved (p = 0.0034) for mice fed a fish-oil enriched diet, compared to the survival curves for mice fed the other 2 diets . The study was repeated by comparing the survival of mice fed a fish-oil enriched diet to those given a corn-oil enriched diet . After 120 h the survival curve for mice fed the fish-oil enriched diet was significantly better compared to the survival curve for mice given the corn-oil enriched diet (p = 0.01) . A fish-oil enriched diet therefore increases survival in mice following Klebsiella pneumoniae infection, whether compared to a standard diet, olive-oil enriched diet or corn-oil enriched diet. Protein Expr Purif, 1997 Dec, 11(3), 257 - 62 Overexpression of arylsulfate sulfotransferase as fusion protein with glutathione S-transferase; Baek MC et al.; A procedure has been developed for the overexpression and purification of milligram quantities of the Klebsiella K-36 arylsulfate sulfotransferase (ASST) . The structural gene was amplified by means of a polymerase chain reaction (PCR) technique and inserted into the plasmid vector pGEX-3X . The plasmid pGEX-100, carrying the Klebsiella K-36 astA structural gene under the control of the Escherichia coli tac promoter, was transformed into the E . coli strain BL21 (DE3) . The ASST was produced in E . coli as a fusion with glutathione S-transferase . Conditions for protein production, isolation on glutathione Sepharose 4B, and Xa cleavage to generate active ASST were developed . The purification yielded approximately 0.7 mg of pure enzyme per liter of bacterial culture . Kinetic analysis of the overexpressed enzyme indicated that it had kinetic properties almost the same as those of the enzyme purified from Klebsiella K-36 cells . The purification procedure was very rapid and is suitable for obtaining considerable amounts of enzyme at a relatively high yield compared with its purifying method from the culture of the Klebsiella K-36 strain. J Appl Toxicol, 1997 Nov-Dec, 17(6), 385 - 90 Glycyrrhizic acid inhibits arylamine N-acetyltransferase activity in Klebsiella pneumoniae in vitro; Lo HH et al.; Glycyrrhizic acid, one of the proposed chemopreventive drugs, was used to inhibit arylamine N-acetyltransferase (NAT) activity in Klebsiella pneumoniae, both in cytosol and intact bacteria . The NAT activity was measured by using high-performance liquid chromatography to assay the amounts of 2-acetyl-aminofluorene and remaining 2-aminofluorene . The NAT activity in K . pneumoniae was inhibited by glycyrrhizic acid in a dose-dependent manner . The cytosol NAT activities were 0.675 +/- 0.028 nmol min(-1) mg(-1) protein for the acetylation of 2-aminofluorene without glycyrrhizic acid and 0.367 +/- 0.008 nmol min(-1) mg(-1) protein with 8 mM glycyrrhizic acid . The NAT activities measured from intact bacteria were 0.308 +/- 0.018 nmol min(-1) 10(-10) colony forming units for the acetylation of 2-aminofluorene without glycyrrhizic acid and 0.236 +/- 0.005 nmol min(-1) 10(-10) colony forming units in the presence of 8 mM glycyrrhizic acid . The inhibition of NAT activity by glycyrrhizic acid was demonstrated to remain for at least 4 h . The apparent Km and Vmax values calculated from cytosol NAT were 1.08 +/- 0.05 mM and 9.09 +/- 0.11 nmol min(-1) mg(-1) protein, respectively, for 2-aminofluorene . In the presence of 8 mM glycyrrhizic acid, the apparent Km and Vmax values were 0.15 +/- 0.01 mM and 0.95 +/- 0.11 nmol min(-1) mg(-1) protein, respectively, for 2-aminofluorene . In intact bacteria, the apparent Km and Vmax values were 1.28 +/- 0.48 mM and 4.08 +/- 1.06 nmol min(-1) 10(-10) colony forming units, respectively, for 2-aminofluorene . However, in the presence of 8 mM glycyrrhizic acid, the apparent Km and Vmax values were 0.67 +/- 0.09 mM and 1.82 +/- 0.37 nmol min(-1) 10(-10) colony forming units, respectively, for 2-aminofluorene . Taking these results together, the NAT activity in K . pneumoniae was inhibited by glycyrrhizic acid both in cytosol and intact bacteria . This study provides the first evidence to demonstrate that glycyrrhizic acid inhibits bacterial NAT activity. Minerva Chir, 1997 Jul-Aug, 52(7-8), 937 - 42 {Totally implantable venous access systems . Analysis of complications}; D'Angelo F et al.; Totally implantable central venous access devices (Port-a-Cath, PaC) allow better treatment of cancer patients, with safe administration of chemotherapeutic agents, and are well accepted by the patients . The aim of the present paper is to analyze the complications of the different implant techniques on the basis of a personal experience of 92 central venous access devices . MATERIAL AND METHODS: A total of 92 PaC (Port-a-Cath, Pharmacia: Celsite Braun) have been implanted in 88 patients between August 1992 and June 1995 for cancer treatment . Age ranged between 19 and 79 years (median 52 years), 56 were male and 32 women . PaC have been implanted by percutaneous cannulation of the subclavian vein, with Seldinger technique, in 34 cases; by venous cutdown respectively on the cephalic vein in 46 cases, the jugular vein in 7 cases, the basilar vein in 4 and the saphenous vein in 1 case . Four patients experienced a double implant . In 84 cases the implant was done under local anesthesia, while in 8 required general anesthesia, during operation for the primary neoplasm . RESULTS: A total of 7 complications were experienced (7.6%, 7/92): 4 sepsis and 3 mechanical . No cases of pnx were observed . Sepsis occurred after 29, 45, 64, 401 days of implantation respectively, and culture demonstrated S . aureus in 2 cases, and E . coli and Klebsiella oxytoca in 1 case each . Mechanical complication comprehends 2 cases of catheter dislodgement and 1 case of port rotation . No complications were noticed in case of implant during surgery for primary cancer (8 cases) . In 7 cases the procedure has been converted from cephalic vein cutdown to percutaneous cannulation of the subclavian vein due to anatomic reasons (13.2%, 7/53) . Five PaC have been explanted for complications . DISCUSSION: On the basis of the personal experience we think that PaC are of easy implant, with few complications and of good acceptance from the patients . We prefer venous cutdown on cephalic vein as implant technique because of avoidance of pnx or bleeding complications . Percutaneous puncture of subclavian vein is useful for implantation during major surgery, because less time consuming, and in case of anatomical anomalies fo the cephalic vein . Basilic vein cutdown has been utilized exclusively for esthetic reason in young people, to avoid the scar in the upper thoracic region . Alternative implant techniques has been employed in special conditions, such as catheter position in the inferior v.cava, or early in our experience (internal jugular vein) . A total of 7 complication have been reported (7.6%), 4 sepsis and 3 mechanical (2 dislodgement, 1 rotation) . Sepsis were not related to implant technique, presenting on day 29, 45, 64 and 401 respectively; all required the explant of the PaC as a treatment . Mechanical complications are related to surgical technique; all required re-exploration with 1 explant and 2 reposition of the PaC . In PaC positioning during surgery for primary cancer (8 cases) no morbidity has been reported . All but the 5 PaC explanted were functioning until patient's need; maximum length reported is 42 months. J Antimicrob Chemother, 1997 Oct, 40(4), 525 - 32 Mechanism and stability of hyperproduction of the extended-spectrum beta-lactamase SHV-5 in Klebsiella pneumoniae; Xiang X et al.; Some isolates of SHV-5 beta-lactamase-producing Klebsiella pneumoniae K2 from a single-strain outbreak of cross-infection produced approximately five-fold more beta-lactamase than others . We investigated three possible genetic mechanisms of this hyperproduction: the presence of a more powerful promoter, an increase in plasmid copy number or an amplification of the gene on a plasmid . No differences between low and high beta-lactamase producers were detected in the promoter region of the SHV-5 beta-lactamase gene, which closely resembled that of SHV-2 . SHV-5 beta-lactamase production was encoded on a low copy number plasmid, but DNA-DNA hybridization with an SHV-specific probe detected a higher gene dose in hyperproducers . The beta-lactamase hyperproduction was unstable on repeated subculture, with a reduction of about 75% after 100 generations . Hyperproducing mutants of a low-producing Klebsiella and its Escherichia coli K-12 transconjugants could be selected in vitro at a frequency of 10(-5) to 10(-6) and these variants had an increased SHV-5 beta-lactamase gene copy number on low copy number plasmids . We conclude that hyperproduction of the extended-spectrum beta-lactamase was caused by gene amplification that could be easily lost or gained in vitro . Since the change to hyperproduction occurred at a high frequency and hyperproducers showed increased resistance to many beta-lactams and beta-lactam/beta-lactamase inhibitor combinations, we suspect that these variants may readily be selected in patients during antibiotic therapy. Eur J Pediatr Surg, 1997 Oct, 7(5), 263 - 6 Characteristics of multisystem organ failure in neonates; Avanoglu A et al.; The records of 45 neonatal deaths in a four year period were reviewed retrospectively . Sixteen patients (35.5%) developed multisystem organ failure (MSOF) . The criteria for pulmonary, hepatic, renal, hematologic, cardiac and microvascular failures were established . The onset of the first organ involvement was calculated in days prior to death . The earliest organ involved was kidney (14.2 +/- 15.1) followed by microvascular (9.4 +/- 7.6), hematologic (84 . +/- 10.1), liver (6.8 +/- 6.7), lung (6.3 +/- 6.6) and cardiac (6.0 +/- 8.7) failure . Blood culture analyses revealed 5 patients with culture-positive sepsis . Yeast was the leading septic agent (n = 3) followed by Klebsiella pneumoniae (n = 2), Pseudomonas sp . (n = 1) and E . coli (n = 1) . The first organ involvement was noted at 17.6 +/- 23.2 days . We concluded that the sequence of neonatal MSOF is different from that of adults, yet the inciting events are not clear-cut . Lung, which is the first organ involved in adults, seems to be a lately involved organ in neonates. Am J Perinatol, 1997 Oct, 14(9), 519 - 21 Ilio-psoas abscess in a neonate; Andreou A et al.; A full-term, small-for-gestational-age, neonate was born 4 days after rupture of the membranes . On the 5th day of life, she developed sepsis due to Klebsiella pneumoniae . On the 18th day of life, the right hip was noted swollen with limited range of motion, but it was painless on passive movements . Ultrasonography revealed abscess of the right ilio-psoas muscle with normal appearance of the right hip joint . Surgical incision and drainage and antibiotic administration resulted in a gradual full recovery . Ultrasonography can confirm the diagnosis of this exceptional clinical entity in neonates, which is difficult to differentiate from septic arthritis of the hip. Appl Microbiol Biotechnol, 1997 Oct, 48(4), 522 - 7 Influence of phosphate on rhamnose-containing exopolysaccharide rheology and production by Klebsiella I-714; Farres J et al.; Physiological conditions enhancing rhamnose-containing polysaccharide synthesis by Klebsiella I-714 were studied in batch culture (0.3-l and 2-l bioreactors) . The four carbon sources tested, sucrose, sorbitol, Neosorb and Cerelose, allowed exopolysaccharide production . Larger amounts of polymer were produced when high carbon/nitrogen ratios and complex nitrogen sources were used . Exopolysaccharide synthesis was greatest at 30 degrees C, which was a suboptimal growth temperature . A reduction in the phosphate content of the medium enhanced rhamnose-containing polysaccharide production . When the initial carbon source concentration was augmented, byproducts other than exopolysaccharide were formed . Rhamnose-containing polysaccharide rheology can be modulated by changing the phosphate content of the medium. Cytometry, 1997 Nov 1, 29(3), 267 - 72 Rapid discrimination of bacterial species with different ampicillin susceptibility levels by means of flow cytometry; Walberg M et al.; An in vitro model for flow cytometric detection of heterogeneous drug response in exponentially growing Escherichia coli and Klebsiella pneumoniae was studied to evaluate the potential of this technology for rapid antibiotic susceptibility testing in polymicrobial samples . The cells, which exhibited 80-fold difference in in ampicillin susceptibility, were cultivated in the presence of ampicillin at a concentration equaling 1 MIC of the low-susceptibility strain (E . coli) . Prior to flow cytometric analysis, the cells were fixed in 70% ethanol and stained with a DNA-specific dye . After 1 h of ampicillin incubation, the light scattering and fluorescence intensities of the susceptible cells increased 4.3-fold and 5-fold, respectively, but remained about constant for the resistant cells . The corresponding cell number increase for the resistant and the susceptible cells was 9.4 and 1.7, respectively . The two strains could be distinguished in the histograms on the basis of their light scattering and fluorescence intensities and by cell number . With an incubation of up to 3 h, the light scattering and fluorescence intensities of the susceptible cells grew stronger, at the expense of cell number . By combination of histograms, the discrimination of different cell populations could be improved . The results demonstrate the ability of flow cytometry to discriminate between species in heterogeneous cultures and suggest the potential of the technique for rapid assessment of bacterial susceptibility . However, the present results are preliminary, and the application to biological liquids and clinical samples has to be demonstrated further. Gene, 1997 Oct 1, 198(1-2), 111 - 3 Evolutionary relationship among rfb gene clusters synthesizing mannose homopolymer as O-specific polysaccharides in Escherichia coli and Klebsiella; Sugiyama T et al.; In order to clarify the evolutionary relationship among rfb gene clusters synthesizing mannose homopolymer as O-specific polysaccharides in Escherichia coli and Klebsiella, we studied the DNA sequence of the boundary region between the rfb and his genes in a series of strains possessing mannose homopolymer as O-specific polysaccharide . All had a characteristic gene organization carrying no gene between the rfb and his genes . Further, the recombination event was suggested to occur at the same site of the hisI gene in those strains . It was suggested that there was a close evolutionary relationship among rfb gene clusters synthesizing mannose homopolymer as O-specific polysaccharide in E . coli and Klebsiella. J Med Microbiol, 1997 Nov, 46(11), 921 - 6 Molecular epidemiology of a large outbreak of multiresistant Klebsiella pneumoniae; Weller TM et al.; An outbreak of multiresistant Klebsiella pneumoniae has continued in the Grampian Region of Scotland since 1992 . The organism, which generally produces an extended-spectrum beta-lactamase (ESBL), has spread to several hospitals and nursing homes . DNA from 80 possible outbreak isolates was digested with the restriction endonucleases XbaI and SpeI, and the patterns obtained by pulsed-field gel electrophoresis were compared . Restriction patterns of 79 of the isolates were found to be highly similar with both restriction enzymes, whereas one isolate was unrelated . The outbreak isolates were divided into six subtypes with SpeI and 16 subtypes with XbaI . These subtypes were independent of antibiotic susceptibility pattern, date of isolation and ward of origin, but the XbaI subtype did correlate with the SpeI subtype . It was concluded that the Klebsiella isolates of this outbreak were clonally related. Epidemiol Infect, 1997 Oct, 119(2), 135 - 42 Klebsiella meningitis in Taiwan: an overview; Tang LM et al.; Klebsiella infection has been considered to be an uncommon cause of meningitis . To determine its incidence and clinical features, we reviewed the microbiologic records of cerebrospinal fluid (CSF) and blood cultures and the medical records of patients with bacterial meningitis admitted between 1981 and 1995 . Klebsiella meningitis was diagnosed in 79 patients with 83 episodes . All patients had klebsiella isolated from CSF and/or blood and typical symptoms and signs of acute bacterial meningitis . Of these, 74 were over 16 years of age and 2 of the 5 children were infants . There was an increased prevalence rate of klebsiella meningitis after 1986 . Of the 83 episodes, only 9 occurred between 1981 and 1986, accounting for 7.8% of 115 cases with CSF and/or blood culture-proven acute bacterial meningitis, whereas in 1987-95, there were 74 episodes accounting for 17.7% of 419 bacteriologically proven cases . K . pneumoniae accounted for 69 episodes, K . oxytoca, 11 episodes and K . ozaenae, 3 episodes . Male gender, diabetes mellitus and liver cirrhosis were commonly associated with K . pneumoniae meningitis . Neurosurgical procedures were frequently associated with K . oxytoca meningitis . All three patients with K . ozaenae meningitis had a primary disease of the nasopharyngeal pathway . The mortality rate due to K . pneumoniae was 48.5%, K . oxytoca, 10% and K . ozaenae, 0% . In patients with K . pneumoniae meningitis, poor prognostic factors included age over 60 years, diabetes mellitus, bacteremia and severe neurological deficits on the first day of treatment. Biosci Biotechnol Biochem, 1997 Oct, 61(10), 1729 - 33 A protein factor is essential for in situ reactivation of glycerol-inactivated adenosylcobalamin-dependent diol dehydratase; Mori K et al.; The adenosylcobalamin-dependent diol dehydratase of Klebsiella oxytoca undergoes suicidal inactivation by glycerol during catalysis involving irreversible dissociation of the Co-C bond of the coenzyme . The glycerol-inactivated holoenzyme in permeabilized cells (in situ) of E . coli harboring a plasmid containing the diol dehydratase genes and their flanking regions was rapidly reactivated in the presence of free AdoCbl, ATP, and Mg2+ . beta,gamma-Methylene ATP was not able to replace ATP . Inactive complexes of the enzyme with aqCbl, CN-Cbl, and PeCbl were activated in situ in the presence of AdoCbl, ATP, and Mg2+, but the complex with AdePeCbl was not . These results suggest that the inactivated holoenzyme is reactivated in situ in the presence of ATP and Mg2+ by exchange of the inactivated coenzyme lacking the adenine moiety for free intact AdoCbl . The in situ reactivation was also observed when an analog lacking the alpha-ribose moiety of the nucleotide loop was used as coenzyme . The results with a recombinant E . coli strains carrying a deletion mutant plasmid demonstrate that certain protein(s) encoded by the 3'-flanking region of the diol dehydratase genes are essential for the in situ reactivation of inactivated diol dehydratase. J Infect, 1997 Sep, 35(2), 185 - 8 Ruptured abdominal aorta secondary to psoas muscle abscess due to Klebsiella pneumoniae in an alcoholic; Sugawara Y et al.; An alcoholic patient with low back pain and Klebsiella pneumoniae septicaemia is reported . Computed tomography revealed abdominal aortic rupture associated with a psoas abscess . Aortic ligation above and below the rupture site and an axillo-femoral bypass were performed, but the patient died on the first postoperative day . Alcoholism is a common underlying disease in K . pneumoniae septicaemia and its septic metastasis to the psoas muscle . The prognosis of aortic infection secondary to psoas abscess is very poor once aortic rupture occurs . Prompt abscess drainage following correct diagnosis and arterial reconstruction before aortic rupture are mandatory. Diagn Microbiol Infect Dis, 1997 Sep, 29(1), 39 - 41 High rates of resistance to piperacillin/tazobactam among Escherichia coli and Klebsiella pneumoniae strains isolated in a Greek hospital; Tsakris A et al.; The activities of piperacillin/tazobactam (PTZ) and 10 other beta-lactams were evaluated by a broth microdilution method using the PASCO system in 1078 Escherichia coli and 447 Klebsiella pneumoniae strains isolated from hospitalized patients in a Greek hospital . Overall, 10.5% of the former and 62.6% of the latter strains displayed resistance to PTZ . Most of the PTZ-resistant strains (71.2%) expressed extended spectrum beta-lactamases and were also resistant to broad spectrum cephalosporins and aztreonam . The high rates of PTZ resistance in E . coli and K . pneumoniae and the extensive cross-resistance to other beta-lactams suggest that PTZ should be used with caution in our clinical setting. Acta Otolaryngol Suppl, 1997, 531, 39 - 51 Roxythromycin prevents endotoxin-induced otitis media with effusion in the guinea pig; Sugiura Y et al.; Pharmacological agents that can normalize or enhance the ciliary and mucociliary activity of the tubotympanum should also be able to break the vicious circle of chronic otitis media with effusion (OME) . Roxythromycin (RXM) has been shown to enhance the ciliary activity in vitro and also stimulate the mucociliary activity in vivo and may therefore, when clinically applied, prevent not only occurrence but also recurrence of clinical OME . The present study was designed to discuss the possible preventive effect of RXM on endotoxin-induced OME in the guinea pig . A total of 120 guinea pigs were used . The normal control group was treated with intratympanic injection of 0.1 ml of physiologic saline solution . The saline-control group was treated with oral administration of physiologic saline solution for 14 successive days . The low-dosage group and the high-dosage group were treated with oral administration of 5 and 50 mg/kg of sairei-to for 14 successive days, respectively . Then, the saline-control group, the low-dosage group and the high-dosage group were treated with intratympanic injection of 0.1 ml of lipopolysaccharide solution (100 micrograms/ml) derived from Klebsiella pneumoniae . All 10 animals in the four groups were sacrificed 1, 3, and 7 days after the intratympanic injection, to examine ciliary activity, mucociliary clearance time, and mucosal pathology of the tubotympanum . The saline-control group exhibited middle ear effusions and pathologies similar to human OME . The incidence of middle ear effusions was significantly reduced in the low-dosage and high-dosage groups . Throughout the observation period, the ciliary activity in the tubotympanum was significantly reduced in the saline-control group as compared with that of the normal control group . By contrast, the ciliary activity in the low-dosage and high-dosage groups was not so reduced in the Eustachian tube and the middle ear close to the orifice . Mucociliary clearance time in the low-dosage and high-dosage groups was not different from that in the normal control group throughout the period . The tubotympanum in the saline-control group exhibited mucosal pathologies characteristic of OME in human . By contrast, the low-dosage and high-dosage groups exhibited much milder pathological changes in the tubotympanum than those in the saline-control group . In conclusion, clinical application of RXM could be an effective measure to prevent the occurrence of OME and also the recurrence of the disease, especially in OME-prone individuals. Acta Otolaryngol Suppl, 1997, 531, 21 - 33 The herbal medicine, sairei-to, prevents endotoxin-induced otitis media with effusion in the guinea pig; Sugiura Y et al.; With current pharmacotherapy, otitis media with effusion (OME) is often recurrent and even develops to become chronic . There is now considerable experimental and clinical evidence that the cilia in the tubotympanum play an important part in the prevention of OME . A herbal medicine, sairei-to, has been shown to stimulate the ciliary activity in vitro, and oral administration of the medicine also stimulated the ciliary activity in the tubotympanum rather than physiological states . This study was designed to investigate whether oral administration of sairei-to could prevent experimental OME in the guinea pig . A total of 120 guinea pigs were used . The control group was treated with intratympanic injection of 0.1 ml of physiologic saline solution . The saline-control group was treated with oral administration of physiologic saline solution for 14 successive days . The low-dosage group and the high-dosage group were treated with oral administration of 120 and 600 mg/kg of sairei-to for 14 successive days, respectively . The saline-control group, the low-dosage group and the high-dosage group were then treated with intratympanic injection of 0.1 ml of lipopolysaccharide solution (100 micrograms/ml) derived from Klebsiella pneumoniae . All 10 animals from the 4 groups were sacrificed 1, 3, and 7 days after the intratympanic injection, to examine ciliary activity, mucociliary clearance time, and mucosal pathology of the tubotympanum . The saline-control group exhibited middle ear effusions and pathologies similar to human OME . The incidence of middle ear effusions in the low-dosage and the high-dosage groups was somewhat reduced compared with the saline-control group . The ciliary activity in the tubotympanum was significantly reduced in the saline-control and low-dosage groups compared with the normal-control group . By contrast, the magnitude of reduction in ciliary activity was much smaller in the high-dosage group . The ciliary activity especially in the Eustachian tube and the middle ear close to the tympanic orifice at 3 and 7 days in the high-dosage group was not significantly different from that in the normal-control group . Mucociliary clearance time in the high-dosage group was not different from that in the normal-control group throughout the observation period . The groups treated with sairei-to, especially the high-dosage group, exhibited much milder pathological changes in the tubotympanum than did the saline-control group . In conclusion, clinical application of sairei-to could be an effective measure to prevent the occurrence of OME and also the recurrence of the disease, especially OME-prone individuals. Arch Pediatr, 1997 Sep, 4(9), 853 - 6 {Acute pyelonephritis and subcapsular hematoma revealing nephroblastoma at the age of 15 years}; Tlili-Graiess K et al.; BACKGROUND: Nephroblastoma' the most common renal tumor in children between 1 and 5 years, occurs rarely in the oldest child . CASE REPORT: A 16-year-old teenager suffered from acute pyelonephritis caused by Klebsiella pneumoniae . Renal ultrasonography showed a left subcapsular hematoma; the CT scan confirmed the finding and also showed renal scarring . However, a second CT scan showed pulmonary nodules suggestive of metastasis, a diagnosis that was confirmed by needle biopsy of pulmonary lesions . Recovery was obtained after chemotherapy and nephrectomy with a 3-year-follow-up . CONCLUSION: This nephroblastoma was particular because its development in an adolescent, its association with acute pyelonephritis and subcapsular hemorrhage. Arch Biochem Biophys, 1997 Nov 1, 347(1), 132 - 40 Heterologous expression, purification, and properties of diol dehydratase, an adenosylcobalamin-dependent enzyme of Klebsiella oxytoca; Tobimatsu T et al.; Recombinant adenosylcobalamin-dependent diol dehydratase of Klebsiella oxytoca overexpressed in Escherichia coli was purified to homogeneity . The enzyme has a low solubility and was extracted from the crude membrane fraction with 1% Brij 35 in a high recovery . Subsequent chromatography on DEAE-cellulose resulted in 4.9-fold purification of the enzyme in an overall yield of 65% . The enzyme thus obtained showed specific activity comparable to that of the wild-type enzyme of K . oxytoca . The apparent molecular weight determined by nondenaturing gel electrophoresis on a gradient gel was 220,000 . The enzyme consists of equimolar amounts of the three subunits with apparent Mr of 60,000 (alpha), 30,000 (beta), and 19,000 (gamma) . Therefore, the subunit structure of the enzyme is most likely alpha2beta2gamma2 . The recombinant enzyme was also separated into components F and S upon DEAE-cellulose chromatography in the absence of substrate . Components F and S were identified as the beta subunit and alpha2gamma2 complex, respectively . Apparent Km for adenosylcobalamin, 1,2-propanediol, glycerol, and 1,2-ethanediol were 0.83 microM, 0.08 mM, 0.73 mM, and 0.56 mM, respectively . The three genes encoding the subunits of diol dehydratase were overexpressed individually or in various combinations in Escherichia coli . The alpha and gamma subunits mutually required each other for correct folding forming the soluble, active alpha2gamma2 complex (component S) . Expression of the beta subunit in a soluble, active form (component F) was promoted by coexpression with both the alpha and gamma subunits, probably by coexistence with component S . These lines of evidence indicate that each subunit mutually affects the folding of the others in this heterooligomer enzyme . Proc Natl Acad Sci U S A, 1997 Oct 28, 94(22), 12145 - 50 Probing the assembly of transcription initiation complexes through changes in sigmaN protease sensitivity; Casaz P et al.; The alternative bacterial sigmaN RNA polymerase holoenzyme binds promoters as a transcriptionally inactive complex that is activated by enhancer-binding proteins . Little is known about how sigma factors respond to their ligands or how the responses lead to transcription . To examine the liganded state of sigmaN, the assembly of end-labeled Klebsiella pneumoniae sigmaN into holoenzyme, closed promoter complexes, and initiated transcription complexes was analyzed by enzymatic protein footprinting . V8 protease-sensitive sites in free sigmaN were identified in the acidic region II and bordering or within the minimal DNA binding domain . Interaction with core RNA polymerase prevented cleavage at noncontiguous sites in region II and at some DNA binding domain sites, probably resulting from conformational changes . Formation of closed complexes resulted in further protections within the DNA binding domain, suggesting close contact to promoter DNA . Interestingly, residue E36 becomes sensitive to proteolysis in initiated transcription complexes, indicating a conformational change in holoenzyme during initiation . Residue E36 is located adjacent to an element involved in nucleating strand separation and in inhibiting polymerase activity in the absence of activation . The sensitivity of E36 may reflect one or both of these functions . Changing patterns of protease sensitivity strongly indicate that sigmaN can adjust conformation upon interaction with ligands, a property likely important in the dynamics of the protein during transcription initiation. Eur J Immunol, 1997 Sep, 27(9), 2123 - 32 Qa-1 interaction and T cell recognition of the Qa-1 determinant modifier peptide; Cotterill LA et al.; The peptide-binding properties of the nonclassical major histocompatibility complex (MHC) class 1b molecule Qa-1 were investigated using a transfected hybrid molecule composed of the alpha 1 and alpha 2 domains of Qa-1b and the alpha 3 domain of H-2Db . This allowed the use of a monoclonal antibody directed against H-2Db whilst retaining the peptide-binding groove of Qa-1b . By comparison with classical MHC class I molecules, intracellular maturation of the chimeric molecule was inefficient with weak intracellular association with beta 2-microglobulin . However, at the cell surface the hybrid molecules were stably associated with beta 2-microglobulin and were recognized by cytotoxic T lymphocyte (CTL) clones specific for the Qa-1b-presented peptide Qdm (AMAPRTLLL) . A whole-cell binding assay was used to determine which residues of Qdm were important for binding to Qa-1b and CTL clones served to identify residues important for T cell recognition . Substitutions at position 1 and 5 did not reduce the efficiency of binding and had little effect on CTL recognition . In contrast, substitutions at position 9 resulted in loss of MHC class I binding . Mass spectrometric analysis of peptides eluted from immunopurified Qa-1b/Db molecules indicated that Qdm was the dominant peptide . The closely related peptide, AMVPRTLLL, which is derived from the signal sequence of H-2Dk, was also present, although it was considerably less abundant . The mass profile suggested the presence of additional peptides the majority of which consisted of eight to ten amino acid residues . Finally, the finding that a peptide derived from Klebsiella pneumoniae can bind raises the possibility that this non-classical MHC class I molecule may play a role in the presentation of peptides of microorganisms. Plasmid, 1997, 38(2), 97 - 105 Characterization of the replication and mobilization regions of the multiresistance Klebsiella pneumoniae plasmid pJHCMW1; Dery KJ et al.; A 2.4-kb EcoRI fragment including the replication and origin of transfer regions of the Klebsiella pneumoniae multiresistance plasmid pJHCMW1 has been cloned and sequenced . The isolated replication region was sufficient for stable maintenance of the plasmid and shares homology with RNA-regulated replicons . Homology was highest with the replication region of the plasmid p15A . Incompatibility experiments, however, determined that pJHCMW1 is compatible with pACYC177, a plasmid harboring the p15A replicon . Differences in their RNA I nucleotide sequences may account for their compatibility . A mobilization origin was also found in the 2.4-kb EcoRI pJHCMW1 DNA fragment analyzed . Conjugation experiments showed that although non-self-transmissible, the recombinant clone including the 2.4-kb EcoRI pJHCMW1 fragment could be mobilized in the presence of the helper plasmid pRK2073. Jpn J Antibiot, 1997 Aug, 50(8), 704 - 10 {Combination effect of fosfomycin to beta-lactam, aminoglycoside, and macrolide antibiotics against clinical isolates of Klebsiella pneumoniae}; O'Hara K et al.; It is well known that fosfomycin (FOM) shows the combination effects with some other antibiotics . Suck effects have not been known against Klebsiella pneumoniae, however . In this report, combination effects of FOM with beta-lactam, aminoglycoside, and macrolide antibiotics were investigated against clinical isolates of both FOM-susceptible and resistant Klebsiella pneumoniae . FOM had synergistic activities with beta-lactams such as ampicillin (ABPC) and cefminox (CMNX), and macrolide antibiotics as erythromycin (EM) and midecamycin (MDM) against all strains tested . Among beta-lactams, penicillin V showed synergistic actions against FOM-susceptible strain of Tf341A and additive actions against FOM-resistant 3 strains with FOM . Pheneticillin was synergistic with FOM against FOM-highly susceptible strain Tf341A and the additive or nearly synergistic effects against other strains . FOM with amoxicillin was synergistic against Tf341A, and FOM-resistant strain of Tf170B and additive against other strains . While the activities of combination of FOM with kanamycin or dibekacin against FOM-susceptible 2 strains were additive, those with amikacin were synergistic . Five different aminoglycosides tested showed antagonistic activities with FOM against 3 FOM-resistant strains . From these results, FOM appears to be clinically useful in treating FOM-susceptible and resistant strains of Klebsiella pneumoniae in combination of 4 antibiotics such as ABPC, CMNX, EM, and MDM. J Bacteriol, 1997 Oct, 179(19), 6014 - 9 Substrate recognition domains as revealed by active hybrids between the D-arabinitol and ribitol transporters from Klebsiella pneumoniae; Heuel H et al.; Two new genes, dalT and rbtT, have been cloned from the dal operon for D-arabinitol and the rbt operon for ribitol uptake and degradation, respectively, in Klebsiella pneumoniae 1033-5P14, derivative KAY2026 . Each gene codes for a specific transporter which, based on sequence data, belongs to a large family of carbohydrate transporters which constitutes 12 transmembrane helices . DalT and RbtT show an unusually high similarity (86.2% identical residues for totals of 425 and 427 amino acids, respectively) . This allowed the construction of DalT'-Rbt"T and RbtT'-Dal'T crossover hybrids by using a natural restriction site overlapping Met202 . This site is located within the large cytoplasmic loop which connects the putative helices 6 and 7 and in particular the amino- and the carboxy-terminal halves of the transporters . Both hybrids have close to normal transport activities but essentially the substrate specificities and kinetic properties of the amino-terminal half . This result localizes essential substrate binding and recognition sites to the amino-terminal halves of the proteins in this important class of carbohydrate transporters. Biochem J, 1997 Sep 15, 326 ( Pt 3), 637 - 40 Nitrogenase of Klebsiella pneumoniae: kinetics of formation of the transition-state complex and evidence for an altered conformation of MoFe protein lacking a FeMoco centre; Yousafzai FK et al.; We have investigated the kinetics of inactivation of Mo-nitrogenase isolated from Klebsiella pneumoniae when it forms an inhibited putative transition-state complex on incubation with ADP and AlF4- . In the presence of excess Kp2 (Fe protein of the Mo-nitrogenase of K . pneumoniae), the kinetics were found to depend on the Mo content of Kp1 (the MoFe protein of Mo-nitrogenase of K . pneumoniae) . The residual nitrogenase activity versus time of incubation using Kp1 preparations containing integral, i.e . one or two Mo atoms per molecule of Kp1, were essentially monophasic, but significantly different rates of inactivation were observed . In contrast, the progress curves for preparations of Kp1 with non-integral Mo content were biphasic, suggesting the presence of two discrete catalytically active species of Kp1 . The best fit to the observed data was obtained with a two-exponential expression, the amplitude of which was consistent with the Mo content, provided that the fast phase of the reaction was assigned to a Kp1 species containing one, and the slow phase to a species containing two Mo atoms per alpha2beta2 tetramer . This analysis provides the first evidence for the existence of a catalytically active Kp1 species containing a single Mo atom . These data also indicate that MoFe protein which does not have all FeMoco binding sites occupied has an altered conformation compared with a fully loaded protein, and that the Fe protein reacts with these conformations at different rates to form the stable, but inhibited transition-state complex. Southeast Asian J Trop Med Public Health, 1997 Mar, 28(1), 218 - 22 Cockroaches from urban human dwellings: isolation of bacterial pathogens and control; Vythilingam I et al.; A study was carried out to determine the distribution of cockroaches in two different housing areas with central sewerage or individual septic tanks in an urban area in Kuala Lumpur, Malaysia . Six species of cockroaches were present and of these Periplaneta americana and Periplaneta brunnea were found in greater abundance . Seventeen species of bacteria were isolated and of these Escherichia coli and Klebsiella p . pneumoniae were isolated in greatest numbers . Control measures carried out using lambda cyhalothrin showed that there was no significant difference between treated and control sites. Heart Lung, 1997 Sep-Oct, 26(5), 413 - 7 Klebsiella pneumoniae pneumonia; Prince SE et al.; Klebsiella pneumoniae is an uncommon cause of community-acquired pneumonia except in alcoholics . Klebsiella may mimic pulmonary reactivation tuberculosis because it presents with hemoptysis and cavitating lesions . Klebsiella pneumoniae is a difficult infection to treat because of the organism's thick capsule . Klebsiella is best treated with third- and fourth-generation cephalosporins, quinolones, or carbapenems . Monotherapy is just as effective as a combination treatment in Klebsiella pneumoniae because newer agents are used . In the past, older agents with less anti-Klebsiella activity were needed for effective treatment . The patient we present was initially thought to have pulmonary tuberculosis, and when found to have Klebsiella pneumoniae, the suggested treatment was monotherapy with ceftriaxone . The patient was treated parenterally initially, and then was treated for 3 weeks with oral ofloxacin. Drugs, 1997, 54 Suppl 1, 33 - 6 Pharmacology of ribosomal immunotherapy; Clot J; The use of immunostimulating drugs is one way to intervene in the immune system . Many of these agents are of bacterial origin and most are able to stimulate the nonspecific immune response by acting on polymorphonuclear cells (PMNs) and macrophages . Ribosomal immunotherapy ('Ribomunyl') contains both proteoglycans from Klebsiella pneumoniae and ribosomes from 4 different bacterial strains . It can stimulate not only macrophages but also specific antibody production . 'Ribomunyl' has been shown to stimulate many of the functions of PMNs, specifically the formation of oxygenated free radicals, chemotaxis and adhesion . The effect of 'Ribomunyl' immunostimulant on the properties of macrophages is of special interest, as these cells participate in both the nonspecific immune response (phagocytosis, proinflammatory cytokine production) and the specific immune response (antigen processing and presentation, lymphocyte proliferation) . 'Ribomunyl' has been shown to increase the production of many cytokines {interleukin (IL)-1, IL-6, IL-8, tumour necrosis factor-alpha and colony-stimulating factor}, leading to the activation of the cytokine network . 'Ribomunyl' was also able to stimulate natural killer cells involved in viral immunity . Because of the presence of ribosomes from 4 frequently encountered bacterial strains, 'Ribomunyl' has specific immunostimulant properties . This has been clearly demonstrated in animals and humans, where specific antibody-forming B cells were found in the tonsils after oral administration . However, specific T-cell response has not been reported, suggesting that 'Ribomunyl' could act directly on B cells such as T-cell-independent bacterial antigens. Curr Microbiol, 1997 Oct, 35(4), 195 - 200 Ibuprofen inhibits arylamine N-acetyltransferase activity in the bacteria Klebsiella pneumoniae; Chung JG et al.; Ibuprofen, one of the nonsteroidal anti-inflammatory drugs, inhibited arylamine N-acetyltransferase activity of Klebsiella pneumoniae both in vitro and in vivo . The NAT activities of Klebsiella pneumoniae were inhibited by ibuprofen in a dose-dependent manner both in vitro and in vivo . In vitro, the NAT activity was 0.675 +/- 0.028 nmol/min/mg of protein for the acetylation of 2-aminofluorene . In the presence of 8 mM ibuprofen, the NAT activity was 0.506 +/- 0.002 nmol/min/mg of protein for the acetylation of 2-aminofluorene . In vivo, the NAT activity was 0.279 +/- 0.016 nmol/min/10(10) colony forming units (CFU) for the acetylation of 2-aminofluorene . In the presence of 8 mM ibuprofen, the NAT activity was 0.228 +/- 0.008 nmol/min/10(10) CFU for the acetylation of 2-aminofluorene . The inhibition of NAT activity by ibuprofen was shown to persist for at least 4 h . For in vitro examination, the values of apparent Km and Vmax were 1.08 +/- 0.05 mM and 9.17 +/- 0.11 nmol/min/mg of protein, respectively, for 2-aminofluorene . However, when 8 mM of ibuprofen was added to the reaction mixtures, the values of apparent Km and Vmax were 1.19 +/- 0.01 mM and 6.67 +/- 0.11 nmol/min/mg of protein, respectively, for 2-aminofluorene . For in vivo examination, the values of apparent Km and Vmax were 1.24 +/- 0.48 mM and 4.18 +/- 1.06 nmol/min/10 x 10(10) CFU, respectively, for 2-aminofluorene . However, when 8 mM of ibuprofen was added to the culture, the values of apparent Km and Vmax were 0.95 +/- 0.29 mM and 2.77 +/- 0.37 nmol/min/mg protein, respectively, for 2-aminofluorene, respectively . This report is the first finding of ibuprofen inhibition of arylamine N-acetyltransferase activity in a strain of Klebsiella pneumoniae. J Biotechnol, 1997 Aug 11, 56(2), 135 - 42 Enzymatic evidence for an involvement of pyruvate dehydrogenase in the anaerobic glycerol metabolism of Klebsiella pneumoniae; Menzel K et al.; Stoichiometric analysis of pathways involved in anaerobic bioconversion of glycerol by Klebsiella pneumoniae revealed that enzyme(s) in addition to pyruvate formate-lyase (PFL) must be involved in pyruvate decarboxylation . In this work, enzymatic evidence is presented that confirmed a simultaneous involvement of pyruvate dehydrogenase complex (PDH) and excluded the presence of pyruvate:ferredoxin oxidoreductase in this anaerobic bioprocess . The in vitro PDH activity of cell extract from continuous culture was found to be strongly affected by the substrate (glycerol) concentration in medium and cell growth rate (dilution rate) . It increases with increasing glycerol concentration and correlates well with the specific substrate uptake rate at different dilution rates in a kind of saturation function . At a similar substrate uptake rate, it decreases with cell growth rate . The in vitro activity of PDH is much higher than its in vivo activity calculated from the pathway stoichiometry but comparable to the calculated in vivo activity of PFL. Zh Mikrobiol Epidemiol Immunobiol, 1997 May-Jun, (3), 73 - 6 {The fractional composition of hydroxylamine preparations and of the lipopolysaccharide of a vaccinal strain of Klebsiella pneumoniae}; Miriasova LV et al.; The fractional composition of the vaccine preparation obtained by the hydroxylamine treatment of K . pneumoniae 204 cells was studied with the use of gel chromatography in sepharose CL-6B . The preparation was shown to form 2 peaks, only the first high molecular peak being serologically active . The method for the treatment of the culture, obtained directly from a fermenter and stored in a frozen state before treatment, was proposed . In the technology of the manufacture of Klebsiella vaccine preparation the possibility of replacing dialysis with gel chromatography was shown. Clin Diagn Lab Immunol, 1997 Sep, 4(5), 550 - 5 O-antigen seroepidemiology of Klebsiella clinical isolates and implications for immunoprophylaxis of Klebsiella infections; Trautmann M et al.; To provide a database for the development of an O-antigen-polysaccharide-containing vaccine against Klebsiella spp., we examined the O-antigen seroepidemiology of 378 Klebsiella clinical isolates collected prospectively in two university centers . Strains were typed by competitive enzyme-linked immunosorbent assay with rabbit antisera specific for serogroups O1 to O12 and monoclonal antibodies (MAbs) specific for serogroups O1, O2ab, O2ac, and the genus-specific core antigen . The numbers of isolates (percentages) of individual O serogroups were as follows: 148 (39.2) for serogroup O1, 40 (10.6) for serogroup O2ab, 4 (1.1) for serogroup O2ac, 89 (23.6) for serogroup O3, 2 (0.5) for serogroup O4, 32 (8.5) for serogroup O5, none for serogroups O7, O9, and O12, and 21 (5.6) for serogroup O11 . Forty-two (11.1) of the strains were non-O-typeable . O-serogroup distributions were virtually identical between isolates from invasive infections and those from noninvasive infections or colonizations . A vaccine containing the O-specific polysaccharides of serogroups O1, O2ab, O3, and O5 would cover 82% of clinically occurring O-antigen specificities . Three hundred thirty-eight of 378 isolates (89.4%) reacted with the genus-specific MAb V/9-5, which recognizes an epitope of the outer core region of Klebsiella lipopolysaccharide . Antibodies directed against this epitope may represent a further alternative for O-antigen-targeted immunoprophylaxis of Klebsiella infections . These data support further experimental investigations on the protective potential of O-antigen-based vaccines and/or hyperimmune globulins in Klebsiella infection. Eur J Surg, 1997 Aug, 163(8), 591 - 6 Comparative trial of four antibiotic combinations for perforated appendicitis in children; Ciftci AO et al.; OBJECTIVE: To compare the therapeutic efficacy of four antibiotic regimens: penicillin, tobramycin, and clindamycin; penicillin, tobramycin, and ornidazole; piperacillin alone; and ceftriaxone and ornidazole in the treatment of children operated on for perforated appendicitis . DESIGN: Prospective randomised study . SETTING: Teaching hospital, Turkey . SUBJECTS: 200 patients aged between 1 and 16 years treated from December 1991 to December 1995 who were randomly assigned to one of four groups each consisting of 50 patients . INTERVENTIONS: Preoperative antibiotics given intravenously, peritoneal drainage by Penrose drains without irrigation, appendicectomy with the inversion of the stump by a purse string, taking peritoneal swabs, and primary skin closure . MAIN OUTCOME MEASURES: Comparability of the groups, duration of fever, leucocytosis, antibiotic treatment, stay in hospital, nasogastric intubation, and drainage, as well as results of cultures and complications . RESULTS: There were no significant differences between the groups for any variable studied . The predominant bacterial species were Escherichia coli, Klebsiella spp, Pseudomonas spp, Fusobacteria, and Peptostreptococci which were appropriately covered by all the antibiotic regimens . Fourteen patients had complications including wound infections (n = 10), prolonged ileus (n = 2) and intra-abdominal abscess (n = 2) all of which were treated conservatively . There was no mortality and no major complications . All regimens had the same clinical and bacteriological efficacy . CONCLUSION: There is no gold standard for antimicrobial chemotherapy in perforated appendicitis . Different antibiotic combinations or a single broad spectrum antibiotic, which include both aerobic and anaerobic coverage, can safely be used in children with perforated appendicitis. Zhonghua Min Guo Xiao Er Ke Yi Xue Hui Za Zhi, 1997 Jul-Aug, 38(4), 282 - 7 Clinical experience with early enteral feeding in very-low-birth-weight infants; Wang LY et al.; The primary objective of this study was to evaluate the safety and benefit of early enteral feeding in very-low-birth-weight (VLBW) infants without parenteral nutrition . Weight gain, feeding intolerance, nosocomial infection rate and a postnatal growth curve were recorded for 61 VLBW premature infants who were admitted to the Neonatal Intensive Care Unit of Mackay Memorial Hospital from September 1, 1995 to February 28, 1997 . Nine infants were unable to complete the study and three were excluded because of severe bronchopulmonary dysplasia; therefore only 49 infants could be evaluated . They were divided into two groups based on birth weight: 1001 gm to 1250 gm (Group A, mean birth weight 1153 +/- 64 gm, mean gestational age 29.0 +/- 2.2 weeks), and less than or equal to 1000 gm (Group B, mean birth weight 911 +/- 82 gm, mean gestational age 27.1 +/- 1.5 weeks) . They received breast milk or premature formula by intermittent nasogastric or continuous nasogastric feeding . Growth was followed over the first 30 postnatal days . Group A reached 100 kcal/kg/day of enteral feeding at a mean age of 17 days as compared with a mean age of 20 days for group B . Infants regained their birth weight at 20 and 25 days in Groups A and B, respectively . By the 30th postnatal day, weight gain exceeded birth weight by 218.2 +/- 143.1 gm and 95.3 +/- 81.5 gm in groups A and B respectively . No definite episodes of necrotizing enterocolitis (NEC) developed . Two cases of Escherichia coli sepsis and one of Klebsiella sepsis occurred . The conclusion was that early enteral feeding in very-low-birth-weight infants does not increase the risk of NEC . It was also demonstrated that enteral feeding alone can produce biphasic postnatal growth curves in very-low-birth-weight infants . Although early enteral feeding was well tolerated in the study infants, the occurrence of feeding intolerance in some (36%) would suggest that additional parenteral nutrition may benefit some infants until full enteral feeding can be achieved. Pathol Biol (Paris), 1997 May, 45(5), 347 - 56 {In vivo pharmacokinetic of amikacin and its pharmacodynamic in combination with cefepime, cefpirome and meropenem in an in vitro/ex vivo micropig model}; Elkhaili H et al.; Three female Yucatan micropigs were included and received a single dose of amikacin (15 mg/kg) by short infusion (30 min) combined either with a single dose of cefepime or cefpirome (30 mg/kg/12 h) or meropenem (7 mg/kg/8 h) . The beta-lactams were administered either by intravenous intermittent injection or by continuous infusion . The mean elimination half-life and clearance value of amikacin were 1.88 h and 2.15 ml/min.kg-1 respectively . These pharmacokinetic parameters were similar to those obtained in man (t1/2 = 2,42 h et Cl = 1,61 ml/min kg-1) . Furthermore, they were not affected by coadministration of cefepime, cefpirome and to meropenem . While resistant to cefepime, cefpirome and amikacin, Klebsiella pneumoniae producing ESBL was susceptible to combination of these cephalosporins with amikacin in an in vitro/ex vivo micropig model . For the six dosage regimens used in this study, the killing activities were similar and resulted in at least 4 log decrease at 6 h after drug exposure . For antimicrobial combination consisting of bolus dosing of amikacin plus continuous infusion of cefepime or cefpirome, the 12 h serum bactericidal titers (SBTs) were 1:8 for cefepime and 1:2 for cefpirome dosage regimen . When each drug administered intermittently, the 12 h SBTs were 1:4 for cefepime and 1:2 for cefpirome . The 8 h SBTs for dosing schedule containing meropenem combined with amikacin were 1:4 and 1:16 after 30 min short infusion and continuous infusion respectively . In conclusion, our study showed that the micropig model is a reliable model for pharmacokinetic investigation of amikacin . It was concluded that beta-lactam antibiotics tested with amikacin may be coadministered by using the standard recommended dosing regimen of amikacin . Continuous infusion of beta-lactams combined with once dosing of amikacin seems to be as or more effective than intermittent injection of each drug. J Clin Microbiol, 1997 Sep, 35(9), 2365 - 9 Identification of clinical isolates of indole-positive Klebsiella spp., including Klebsiella planticola, and a genetic and molecular analysis of their beta-lactamases; Liu Y et al.; In a collection of 43 indole-positive Klebsiella clinical isolates, which were initially identified as Klebsiella oxytoca, there were 18 isolates which exhibited a pattern characteristic of extended-spectrum beta-lactamase (ESBL) resistance . This study aimed to confirm their identity by biochemical tests and by PCR and to determine the genetic basis for their resistance to the beta-lactams and broad-spectrum cephalosporins . Chromosomal beta-lactamase genes were analyzed by PCR, and plasmid-mediated beta-lactamase genes were analyzed by conjugation and transformation . There were 39 isolates which grew on melezitose but failed to grow on 3-hydroxybutyrate, confirming them as K . oxytoca . PCR analysis of their beta-lactamase genes divided these isolates into two groups, the bla(OXY-1) group and the bla(OXY-2) group . Each group had beta-lactamases with different isoelectric points; the bla(OXY-1) group had beta-lactamases with isoelectric points at 7.2, 7.8, 8.2, and 8.8, and the more common bla(OXY-2) group had beta-lactamases with pIs at 5.2, 5.4 (TEM-1), 5.7, 5.9, 6.4, and 6.8 . A pI of 5.2 was the most frequently detected and accounted for 59% of all the bla(OXY-2) beta-lactamases . Hyperproduction of clavulanate-inhibited chromosomal beta-lactamases was detected in 17 K . oxytoca isolates, resulting in an ESBL phenotype . K . oxytoca isolates having a plasmid-mediated genetic basis for their ESBL phenotype were not found, confirming that, in K . oxytoca, plasmids are rarely involved in conferring resistance to the newer cephalosporins . Four isolates proved to be isolates of K . planticola in which the beta-lactamase genes failed to react with the primers used in the PCR . One K . planticola isolate contained a transferable plasmid harboring the SHV-5 beta-lactamase gene and showed an ESBL phenotype, while the other non-ESBL K . planticola isolates contained chromosomal beta-lactamases with isoelectric points at 7.2, 7.7, and 7.9 plus 7.2. J Clin Microbiol, 1997 Sep, 35(9), 2191 - 7 Comparison of screening methods for detection of extended-spectrum beta-lactamases and their prevalence among blood isolates of Escherichia coli and Klebsiella spp . in a Belgian teaching hospital; Vercauteren E et al.; Using a set of 33 well-defined extended-spectrum beta-lactamase (ESBL)-producing strains of Escherichia coli and Klebsiella pneumoniae, we compared three screening methods for ESBL detection: (i) a double-disk synergy test, (ii) a three-dimensional test (both the double-disk synergy test and the three-dimensional test were performed with ceftriaxone, ceftazidime, aztreonam, and cefepime), and (iii) the Etest ESBL screen (AB Biodisk, Solna, Sweden), based on the recognition of a reduction in the ceftazidime MIC in the presence of clavulanic acid . In the double-disk test, all four indicator antibiotics scored equally and 31 of the 33 reference strains were recognized . In the three-dimensional test, ceftriaxone was the only satisfactory indicator and 30 ESBL-positive strains were detected by this antibiotic . Both systems produced two false-positive results with cefepime . With the Etest ESBL screen, 15 of 16 TEM-related and 11 of 16 SHV-related ESBL-producing strains scored positive . In 10 cases the clavulanic acid on one end of the strip interfered with the MIC determination for ceftazidime, which was read on the opposite end . This MIC had to be determined with an extra ceftazidime-only strip . No false-positive results were noted . Eighty-six blood isolates of E . coli and Klebsiella species were screened for ESBL expression by the double-disk and three-dimensional tests, both with ceftriaxone . Six strains with suspicious antibiogram phenotypes also gave positive results by the double-disk test . One E . coli strain remained undetected by the three-dimensional test . Identification of the enzymes suspected of being ESBLs by isoelectric focusing (all strains) and DNA sequencing (1 strain) confirmed the screening test results except for one Klebsiella oxytoca strain, which proved to be a hyperproducer of its chromosomal enzyme and which also had a negative Etest score . The five true ESBL producers were all confirmed by the Etest ESBL screen . Pulsed-field gel electrophoresis proved that the E . coli strains were unrelated, but that two of the three K . pneumoniae strains were closely related. J Mol Biol, 1997 Aug 22, 271(3), 342 - 8 KpnAI, a new type I restriction-modification system in Klebsiella pneumoniae; Lee NS et al.; The KpnAI restriction-modification (R-M) system has been identified in Klebsiella pneumoniae strain M5a1 . The restriction gene of KpnAI was first cloned into pBR322 using an r-m+ M5a1 derivative and phage SBS for screening . Subsequently, an adjacent DNA fragment showing modification activity was cloned into pUC19 . A total of 7.2 kb DNA sequencing data revealed three open reading frames, corresponding to hsdR, hsdM and hsdS genes of type I R-M systems . The predicted hsdR, hsdM and hsdS-coded peptides shared 95%, 98% and 44% identity, respectively, with the corresponding peptides of the recently identified StySBLI system, a prototype of the type ID family . This high homology suggests that KpnAI is also a member of the type ID family . The KpnAI system seems to be the first type I system identified in Klebsiella species. J Appl Microbiol, 1997 Aug, 83(2), 166 - 74 Formate and ethanol are the major products of glycerol fermentation produced by a Klebsiella planticola strain isolated from red deer; Jarvis GN et al.; The rumen contents of red deer (Cervus elaphus) were used to isolate bacterial capable of fermenting glycerol . The biochemistry, physiology, morphology and phylogeny of one isolate were studied in detail . The isolate (DR3) was tentatively identified as a strain of the species Klebsiella planticola as based on phenotypic characterization . The data obtained from 16S rRNA sequence analysis showed that the deer rumen iso