J Bacteriol, 1994 Feb, 176(3), 714 - 24 Inhibition of erythromycin synthesis by disruption of malonyl-coenzyme A decarboxylase gene eryM in Saccharopolyspora erythraea; Hsieh YJ et al.; Malonyl-coenzyme A (malonyl-CoA) decarboxylase is widely distributed in prokaryotes and eukaryotes . However, the biological function of this enzyme has not been established in any organism . To elucidate the structure and function of this enzyme, the malonyl-CoA decarboxylase gene from Saccharopolyspora erythraea (formerly Streptomyces erythreaus) was cloned and sequenced . This gene would encode a polypeptide of 417 amino acids . The deduced amino acid sequence matched the experimentally determined amino acid sequences of 25 N-terminal residues each of the enzyme and of an internal peptide obtained by proteolysis of the purified enzyme . This decarboxylase showed homology with aminoglycoside N6'-acetyltransferases of Pseudomonas aeruginosa, Serratia marcescens, and Klebsiella pneumoniae . Northern (RNA) blot analysis revealed a single transcript . The transcription initiation site was 220 bp upstream of the start codon . When expressed in Escherichia coli, the S . erythraea malonyl-CoA decarboxylase gene yielded a protein that cross-reacted with antiserum prepared against S . erythraea malonyl-CoA decarboxylase and catalyzed decarboxylation of {3-14C}malonyl-CoA to acetyl-CoA and 14CO2 . The S . erythraea malonyl-CoA decarboxylase gene was disrupted by homologous recombination using an integrating vector pWHM3 . The gene-disrupted transformant did not produce immunologically cross-reacting 45-kDa decarboxylase, lacked malonyl-CoA decarboxylase activity, and could not produce erythromycin . Exogenous propionate restored the ability to produce erythromycin . These results strongly suggest that the decarboxylase provides propionyl-CoA for erythromycin synthesis probably via decarboxylation of methylmalonyl-CoA derived from succinyl-CoA, and therefore the malonyl-CoA decarboxylase gene is designated eryM . The gene disrupted mutants also did not produce pigments. Microbiology, 1994 Feb, 140 ( Pt 2), 311 - 20 Cloning and expression in Escherichia coli of a Streptomyces coelicolor A3(2) argCJB gene cluster; Hindle Z et al.; From a partial Sau3AI library of Streptomyces coelicolor A3(2) DNA in pIJ916, two hybrid plasmids pGX1 and pGX2 were isolated that complemented S . coelicolor A3(2) or S . lividans arginine auxotrophs . Subcloning DNA from pGX1 in the Escherichia coli expression vector pRK9 containing the Serratia marcescens trp promoter gave rise to one plasmid, pZC2, that complemented E . coli argB, C, E and H auxotrophs, and another, pZC1, that complemented only the first three . The plasmids were markedly unstable in the various complemented hosts, to varying extents; pZC1 was characterized further as providing the stablest host/plasmid combinations . In vitro deletion of part of the vector's trp promoter did not affect complementation of the argB and C auxotrophs, implying that the S . coelicolor A3(2) arg genes may be expressed from their own promoter . The trp promoter-less plasmids included isolates, such as pZC177, that had suffered extensive further deletion without loss of complementing ability . Extracts of an E . coli argE auxotroph carrying pZC177 showed ornithine acetyltransferase activity, indicating that the complementing gene is of the argJ type . The complementation properties of in vitro deletion derivatives of pZC177 indicated the gene order argC-J-B . Part of argC and the upstream region were sequenced; an ORF was identified whose predicted product showed appreciable homology with the E . coli and Bacillus subtilis ArgC polypeptide . Upstream of this ORF a consensus-type promoter and ribosome binding site could be discerned; overlapping its promoter was a sequence with homology to arginine operators in these two other organisms.(ABSTRACT TRUNCATED AT 250 WORDS) Protein Expr Purif, 1994 Feb, 5(1), 37 - 43 A procedure for renaturation and purification of the extracellular Serratia marcescens nuclease from genetically engineered Escherichia coli; Friedhoff P et al.; Overproduction of the extracellular Serratia marcescens nuclease in Escherichia coli results in aggregation and sequestration of a large amount of the protein in inclusion bodies . Only a relatively small amount is secreted into the medium from which it can be purified following established procedures . The cell-associated insoluble protein can be solubilized in 6 M urea after breaking up the cells by sonication . Renaturation is achieved by dilution or dialysis . Subsequent phosphocellulose chromatography yields a homogeneous protein preparation which is shown by a variety of biochemical and biophysical analyses to be indistinguishable from conventionally prepared material . The high yield (> 10 mg/500-ml culture) and the ease of preparation (2 to 3 days) make this an attractive alternative to previously described procedures. Kansenshogaku Zasshi, 1994 Feb, 68(2), 177 - 82 {Bacterial contamination of hair washing liquids}; Amemiya K et al.; To determine the extent of contamination by bacteria of hair-washing shampoo and rinse used professionally at barber shops and hair-dressing saloons, quantitative isolation of bacteria were performed by using a total of 39 samples of shampoo and rinse fluid obtained from 17 facilities . It was found that a maximal number of 1 x 10(7)/ml colony forming units/ml of bacteria were isolated from 60.7% (17 out of 28 samples) of the shampoo and 45.5% (5 out of 11) of the rinse . Gram-negative bacilli were the predominant strains (87.9%) involved in bacterial contamination and the major isolates were Serratia marcescens (43.3%, most frequently isolated), Pseudomonas cepacia, P . fluorescens, P . aeruginosa and Klebsiella pneumoniae, all which are widely recognized as nosocomial-infection causing pathogens . These results indicate hair-washing liquids for professional use are contaminated with a great number of gram-negative bacteria, being possible causes of nosocomial infections, and much attention should be paid to the sanitation and cleanliness of the shampoo and rinse for hair-washing. J Clin Microbiol, 1994 Feb, 32(2), 575 - 7 Nosocomial septicemia caused by Serratia plymuthica; Domingo D et al.; We report a case of nosocomial septicemia in a 79-year-old patient caused by Serratia plymuthica with no evident focus of infection . The patient was treated with gentamicin (40 mg every 8 h) during 10 days; clinical resolution of the infection was obtained after the 10-day treatment period. Epidemiol Infect, 1994 Feb, 112(1), 125 - 31 Aminoglycoside resistance patterns of Serratia marcescens strains of clinical origin; Coria-Jimenez R et al.; Aminoglycoside resistance patterns of 147 Serratia marcescens strains of clinical origin were studied . All strains analysed belonged to three different bacterial populations . The periods of study and the institutions the strains were isolated from correlated significantly with the resistance patterns shown by the strains . The most frequent resistance patterns found were the following: ACC (6')-I at the Hospital Infantil de Mexico (Children's Hospital of Mexico), and ANT (2'') + AAC(6')-I at the Instituto Nacional de Pediatria (INPed or National Institute of Pediatrics) in Mexico City . Furthermore, the isolation frequency of aminoglycoside-sensitive strains decreased remarkably at the INPed over a 12-year period . These results suggest that there has been a selection of Serratia marcescens strains that are very resistant to aminoglycosides. Can J Microbiol, 1994 Feb, 40(2), 120 - 6 Characterization and primary specificity of an extracellular metalloproteinase from Serratia marcescens; Kim N et al.; An extracellular endopeptidase (proteinase) from Serratia marcescens (Serratia marcescens extracellular proteinase, EC 3.4.24.4), purified to homogeneity, was analyzed for enzyme properties . The enzyme has a polypeptide chain molecular mass of 52 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) . The enzyme has an optimal temperature of 40 degrees C and an optimal pH of 7.0 . Enzyme activity was enhanced over two times by the addition of Ca2+ and Mg2+ ions and eliminated almost completely by the presence of 0.2% SDS . The enzyme has broad substrate specificity and contains neither cysteine nor methionine . Low homology was found between the NH2-terminal amino acid sequence of the enzyme of this study and the NH2-terminal sequence of a proteinase from another strain of S . marcescens . Chemical modification with N-bromosuccinimide, 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, and 8-anilino-1-naphthalene sulfonic acid and by photooxidation with methylene blue reduced enzyme activity considerably . The enzyme was shown to have broad peptide bond specificity judging from the contribution of 11 amino acids to the carboxyl side of the peptide bonds hydrolyzed. Nippon Kyobu Geka Gakkai Zasshi, 1994 Jan, 42(1), 95 - 100 {Curative report of post sternotomy mediastinitis due to bacterofungal infection}; Katsumata T et al.; We describe a 72-year-old patient with mediastinitis caused by Methicillin-resistant Staphylococcus aureus (MRSA) and Candida albicans after median sternotomy who was successfully treated with wound closure with pectoral musculocutaneous flap and closed continuous mediastinal irrigation . The irrigation device consisted of two pairs of irrigation and suction tubes which placed upper and lower half of mediastinum respectively, in which high rate irrigation technique (200 ml/h) was employed using 0.01% of Vancomycin hydrochloride as a base agent and additional 0.1% of Povidone-iodine in early phase and 0.01% of Fluconazole in late phase . After 12-days irrigation, the drainage culture turned negative and the wound was healed and tomographically granulated . High rate irrigation with sufficient concentration of antimicrobial agents selected according to each organism sensitivity could eliminate redundant irrigation and contribute to avoid antimicrobial toxication . We experienced also another four cases (two caused by MRSA, two caused by Serratia liquefaciens) treated successfully with this technique . These results led us to believe that continuous mediastinal irrigation technique could be carried out with safe and effectiveness so far as high rate irrigation with low concentration of Povidone-iodine is employed. J Antimicrob Chemother, 1994 Jan, 33(1), 91 - 101 Cefminox: correlation between in-vitro susceptibility and pharmacokinetics and serum bactericidal activity in healthy volunteers; Aguilar L et al.; Plasma concentration of cefminox and serum bactericidal activity against four ATCC strains (Escherichia coli 25992, Klebsiella pneumoniae 13833, Serratia marcescens 8100 and Bacteroides fragilis 25285), were determined over a 24 h period after administration of cefminox 1 and 2 g to six healthy volunteers in a randomized, cross-over, single blind study . The increase observed in the area under the bactericidal curve (AUBC) with the 2 g dose was at least 3.5 times that seen with the 1 g dose for all four test strains and was larger than predicted by the corresponding increase (1.84 times) in the area under the serum concentration versus time curve (AUC); a correlation (r = 0.88, P = 0.0001) between the cefminox concentration and the serum bactericidal titres was, however, observed with all four strains tested . The MBC6h showed a better association with the serum bactericidal titre (P < 0.01) than did the MIC or MBC. Arch Microbiol, 1994, 161(2), 176 - 83 The extracellular nuclease of Serratia marcescens: studies on the activity in vitro and effect on transforming DNA in a groundwater aquifer microcosm; Ahrenholtz I et al.; A quantitative endonuclease assay, which relies on the introduction of single and double strand breaks into supercoiled plasmid DNA, was used to study the activity of the extracellular nuclease of Serratia marcescens SM6 in buffer and in groundwater . The parallel enzyme concentration-dependent production of relaxed and linear plasmid molecules suggests that the nuclease produces single and double strand breaks in duplex DNA . Bovine serum albumin stimulated the nuclease activity towards DNA and RNA and increased the stability of the enzyme against thermal inactivation . The DNase activity at 4 degrees C and 50 degrees C was almost half of that at the optimum temperature (37 degrees C) . The nuclease was active in groundwater, although the specific activity was lower than in buffer . In a groundwater aquifer microcosm, mineral-adsorbed transforming DNA was substantially less accessible to the nuclease than was dissolved DNA . The data suggest that the extracellular nuclease of Serratia marcescens may contribute to DNA turnover in the environment and that adsorption of DNA to minerals provides protection against the nuclease. Res Microbiol, 1994 Jan, 145(1), 17 - 25 Replicon typing of 71 multiresistant Serratia marcescens strains; Llanes C et al.; Replicon typing is the identification of plasmids by hybridization with specific DNA probes which contain the genes involved in plasmid maintenance . This new method has been used to classify plasmids into replicon (rep) groups which can often be correlated with incompatibility (Inc) groups . We studied 71 multiresistant Serratia marcescens strains with 19 rep probes constructed from reference plasmid replicons belonging to known Inc groups . These probes are known to react with enteric bacterial plasmids . However, they did not represent the totality of the thirty known Inc groups . For 52% of the studied strains, plasmids were identified and classified into groups FIB, FIC, FIIA, HI2, L/M, N, B/O, P, W, Y and Com9 . Most (79%) of the plasmid preparations hybridized with a single rep probe, and 21% hybridized with two different probes . Electrophoretic analysis of DNA suggested that double hybridization could result from the presence of either two different Inc plasmids in the same strain (e.g . S37) or one single plasmid with a multireplicon (e.g . S113). Microbiol Immunol, 1994, 38(3), 171 - 5 A function of 40 kDa outer membrane protein in Serratia marcescens; Tada Y et al.; The function of one of the outer membrane proteins of Serratia marcescens was investigated . S . marcescens with an abundant 40 kDa outer membrane protein was induced to form spheroplast at a high rate in an isotonic medium in the presence of calcium, although the spheroplasts were generally fragile in the isotonic environment . The degree of spheroplast induction was correlated to the amount of the 40 kDa protein present in the membrane . In the 40 kDa proteinless mutant strains, the spheroplast induction rate was remarkably decreased . Autoradiography of the outer membrane revealed the presence of a calcium-binding protein as a radioactive band whose position coincided with the 40 kDa protein . These results suggest that the 40 kDa protein has an important role in maintaining the structural integrity of the cell wall against osmotic shock. Arch Microbiol, 1994, 161(5), 414 - 7 Isolation of Serratia marcescens mutants which could overproduce and excrete Escherichia coli alkaline phosphatase and beta-lactamase; Asdornnithee S et al.; Serratia marcescens mutants, which excrete Escherichia coli alkaline phosphatase (APase) encoded by the plasmid-bearing phoA gene, were isolated after mutagenesis by N-methyl-N'-nitro-N-nitrosoguanidine . These mutants produced two to four times as much APase as did the parent strain under a phosphate-limiting condition, and more than 70% of the enzyme was released into the culture medium . In addition, overproduction and excretion of beta-lactamase was observed in these mutants. Microbios, 1994, 79(320), 155 - 61 Effect of various growth conditions on pigmentation of Serratia marcescens; Rjazantseva IN et al.; Prodigiosin biosynthesis in Serratia marcescens depends on growth conditions . In the presence of various concentrations of NaCl, pigment synthesis started later, but up to 4-5% NaCl increased prodigiosin accumulation per biomass unit . Visible light (< 2,000 lux) influenced pigmentation without changing the growth characteristics of the culture . The maximum prodigiosin content in dark and light cultures was observed on days 3-4 and 2-3, respectively . The influence of illumination conditions on pigmentation of S . marcescens was demonstrated . Light directly affects the pigment itself which is synthesized by the culture . Growing cells contain mono and dimer forms of prodigiosin in glycerol, with light influencing these pigments. Klin Lab Diagn, 1994, (4), 41 - 2 {Identification of Serratia marcescens by a modified crystal coating method}; Bazhenov LG; Crystal coating method modified for microbiologic studies is presented . Studies of 230 microorganism strains referred to different species revealed specific features characteristic of S . marcescens crystallogram only: the presence of luxurious whitish formations shaped as ramified branches with air vesicles and small white balls . Detection of these features permit a reliable identification of S . marcescens. Eur J Cardiothorac Surg, 1994, 8(12), 665 - 6 Mycotic pulmonary artery aneurysm following pulmonary artery banding; Kumar RV et al.; A neonate with situs inversus, transposition of the great arteries, ventricular septal defect, criss-cross ventricles and hypoplastic right ventricle underwent pulmonary artery banding at the age of 7 days . The course was complicated by septicaemia and subsequently the development of an aneurysm of the pulmonary artery . Serratia marcessans was grown from the band site . The pulmonary artery aneurysm was resected and the pulmonary artery was repaired . The literature is reviewed with the emphasis on diagnosis, natural history and surgical management. Biochem Biophys Res Commun, 1993 Dec 15, 197(2), 536 - 41 Superoxide dismutase mimetic activities of metal complexes of L-2(2-pyridyl)-1-pyrroline-5-carboxylic acid (pyrimine); Itami C et al.; We previously isolated the strain Z-54 (Serratia marcescens O5:H1) which produces a reddish-violet pigment . The structure of this pigment was confirmed to be that of a peptide complex containing Fe2+ and L-2(2-pyridyl)-1-pyrroline-5-carboxylic acid (pyrimine), as a chromophore . We measured the superoxide dismutase mimetic activities for the pyrimine-metal complexes by xanthine oxidase/nitroblue tetrazolium and cytochrome c methods and found that the pyrimine-Cu2+ (2:1) complex shows the highest activity yet reported (IC50 = 0.11 microM) among the complexes tested . Pyrimine-Cu+, -Fe2+ and -Mn2+ complexes also gave relatively high SOD mimetic activities . ESR spectra observed for pyrimine-Cu2+ (4:1) showed the structure of the Cu(2+)-complex to be tetrahedral and coordinated with four nitrogen atoms . These results support the idea that the pyrimine-metal complexes might be potent SOD mimics. Biotechnol Appl Biochem, 1993 Dec, 18 ( Pt 3), 389 - 99 Characterization of natural and recombinant nuclease isoforms by electrospray mass spectrometry; Pedersen J et al.; Isoforms of natural and recombinant nuclease have been characterized on the basis of their M(r) as determined by electrospray m.s. . The natural nuclease was isolated and purified from Serratia marcescens B10M1 and the recombinant nuclease from Escherichia coli MT102 carrying the plasmid p403-SD2 . The primary structure of each of the isoforms isolated from the nuclease preparations was established by comparing their mass with the known amino acid sequence derived from the nucleotide sequence of the nuc gene . All the preparations were found to be contaminated with the same N-terminal split variants of native nuclease, although the natural nuclease contained much larger amounts of these isoforms than did the recombinant nuclease . The structure of some of the isoforms could be verified by N-terminal sequencing, and nearly all of them by isoelectric focusing. Klin Monatsbl Augenheilkd, 1993 Dec, 203(6), 418 - 22 {Bilateral acanthamoeba keratitis}; Holz FG et al.; BACKGROUND: Given the protracted clinical course, diagnostic difficulties, frequent treatment failures and the increasing incidence, keratitis caused by free-living acanthamoeba represents a clinical challenge . PATIENT AND METHODS: A 37-year-old healthy female who wore gas-permeable contact lenses for ten years developed bilateral keratouveitis, pseudodentritic subepithelial infiltrates and corneal ring ulcers . Cultures were obtained from corneal scrapings, contact lenses and storage container . Medical treatment during the clinical course included propamidine isethionate, miconazole, ketoconazole, polymyxin B, aminoglycoside antibiotics and corticosteroids . Surgical procedures included bilateral penetrating keratoplasty and extracapsular cataract extraction . Each corneal button was examined after chemofluorescent staining with calcofluor white . RESULTS: Klebsiella oxytoca and Serratia marcescens were grown from cultures of the contact lens storage container . Although suspected early in the clinical course repeated cultures from corneal scrapings were negative for acanthamoeba . Despite transient remission, medical therapy including therapy for acanthamoeba could not halt the progression of infection in both eyes . Visual acuity deteriorated to light perception and counting fingers, respectively . Penetrating keratoplasty was performed 12 and 15 months after the onset of symptoms . Histopathological examination allowed identification of acanthamoeba cysts in each button . Because of secondary cataract formation cataract-extraction with intraocular lens implantation was simultaneously performed in the right and subsequently in the left eye . While corneal infiltrates recurred and optic atrophy developed due to secondary glaucoma in the right eye, the left corneal graft has remained clear . CONCLUSIONS: The case-report demonstrates that diagnostic procedures may fail to detect acanthamoeba organisms before obtaining a corneal button for histopathologic examination . Decreased corneal sensation later in the clinical course after initial pain disproportionately related to clinical signs does not exclude the diagnosis of an acanthamoeba keratitis . Medical treatment failure may occur despite early initiation of antiparasitic therapy. Appl Biochem Biotechnol, 1993 Dec, 43(3), 189 - 97 Improvement of culture conditions for L-proline production by a recombinant strain of Serratia marcescens; Masuda M et al.; Serratia marcescens SP511 was previously reported to be an L-proline-producing strain that harbors a recombinant plasmid carrying the mutant type of the proline operon . This strain produced 65 g/L of L-proline in a medium containing 22% sucrose and urea after 5 d of incubation under the conventional culture conditions . We searched for more suitable culture conditions for more abundant L-proline production by SP511 . To improve the supply of a nitrogen source to cells, ammonium was used instead of urea and fed to a culture under control of the pH of the medium . The concentrations of MgSO4 and K2HPO4 were increased, and in addition, sucrose was continuously added to the culture at a final concentration of 32% . Under these conditions, the cell amount was increased twofold over that under the previous conditions and L-proline production reached a maximum of more than 100 g/L after 4 d of incubation. Cornea, 1993 Nov, 12(6), 489 - 92 Suture abscesses after penetrating keratoplasty; Leahey AB et al.; Eighteen suture abscesses that developed after penetrating keratoplasty in 15 patients were reviewed . The time from keratoplasty to the diagnosis of an abscess ranged from 1 to 53 months with a mean of 21.5 months . In 13 of the 18 cases, the patient was taking topical steroids at the time of diagnosis . All were culture-proven bacterial ulcers, except for one case that had a positive Gram's stain, but no growth on culture . The organisms cultured were Staphylococcus epidemidis (six eyes), Streptococcus pneumoniae (five eyes), Sta . aureus (four eyes), Str . viridans (two eyes), Klebsiella oxytoca (one eye), Serratia marcescens (one eye), Moraxella sp (one eye), and Escherichia coli (one eye) . The offending suture was removed in all cases, and the eyes were treated with topical fortified antibiotics (cefazolin and tobramycin) . After treatment, 67% (12 of 18 eyes) had clear grafts, 17% (three of 18 eyes) were scarred, and 16% (three of 19 eyes) had failed grafts . Intensive topical steroid therapy was used when a subsequent graft rejection developed . Retained sutures following corneal transplants can result in sight-threatening infections and should be considered for removal as soon as the wound is well healed. EMBO J, 1993 Nov, 12(11), 4151 - 7 First structure of a snake venom metalloproteinase: a prototype for matrix metalloproteinases/collagenases; Gomis-Ruth FX et al.; Adamalysin II, a 24 kDa zinc endopeptidase from the snake venom of Crotalus adamanteus, is a member of a large family of metalloproteinases isolated as small proteinases or proteolytic domains of mosaic haemorrhagic proteins from various snake venoms . Homologous domains have recently been detected in multimodular mammalian reproductive tract proteins . The 2.0 A crystal structure of adamalysin II reveals an ellipsoidal molecule with a shallow active-site cleft separating a relatively irregularly folded subdomain from the calcium-binding main molecular body composed of a five-stranded beta-sheet and four alpha-helices . The folding of the peptide fragment containing the zinc-binding motif HExxHxxGxxH bears only a distant resemblance to thermolysin, but is identical to that found in astacin, with the three histidines and a water molecule (linked to the glutamic acid) likewise constituting the zinc ligand; adamalysin II lacks a fifth (tyrosine) zinc ligand, however, leaving its zinc ion tetrahedrally co-ordinated . Furthermore, adamalysin II and astacin share an identical active-site basement formed by a common Metturn . Due to their virtually identical active-site environment and similar folding topology, the snake venom metalloproteinases (hitherto called adamalysins) and the astacins (and presumably also the matrix metalloproteinases/mammalian collagenases and the Serratia proteinase-like large bacterial proteinases) might be grouped into a common superfamily with distinct differences from the thermolysin family. J Biochem (Tokyo), 1993 Nov, 114(5), 723 - 31 Characterization of secretory intermediates of Serratia marcescens serine protease produced during its extracellular secretion from Escherichia coli cells; Shikata S et al.; The Serratia marcescens serine protease (SSP; 66 kDa) is synthesized as a precursor (preproSSP; 112 kDa) composed of the NH2-terminal signal peptide of 27 amino acids, the mature protease part and a large COOH-terminal domain . When the SSP gene is expressed in Escherichia coli under the control of the tac promoter, the mature enzyme is excreted into the medium through the outer membrane, whereas preproSSP and two proteins, C-1 (40 kDa) and C-2 (38 kDa), processed from the COOH-terminal domain, are accumulated in the membrane fraction . Although treatment of the intact cells with trypsin caused slight truncation of C-1 and C-2, the main parts of C-1 and C-2, both of which are detected in the outer membrane, were resistant to trypsin, even after the cells had been osmotically shocked . Consistent with this, a high content of beta-sheet structure in C-2 was suggested by marked heat-modifiability, as determined by their electrophoretic mobilities on SDS-polyacrylamide gel . These findings suggest rigid integration of C-1 and C-2 in the outer membrane . Upon induction of the tac promoter, rapid excretion of SSP into the medium was first accompanied by the accumulation of C-1 in the outer membrane, which was followed by conversion of C-1 to C-2 . PreproSSP was not detected during the accumulation of SSP in the medium, but it was gradually accumulated after the accumulation of SSP had reached a plateau . In addition, preproSSP still containing the intact NH2-terminal signal peptide was completely digested with trypsin when added to osmotically shocked cells.(ABSTRACT TRUNCATED AT 250 WORDS) Appl Environ Microbiol, 1993 Oct, 59(10), 3225 - 32 Molecular breeding of a biotin-hyperproducing Serratia marcescens strain; Sakurai N et al.; We previously reported that an acidomycin-resistant mutant of Serratia marcescens Sr41, SB304, and a mutant that was derived from SB304 and was resistant to a higher concentration of acidomycin, SB412, produced 5 and 20 mg of D-biotin, respectively, per liter of a medium containing sucrose and urea (N . Sakurai, Y . Imai, M . Masuda, S . Komatsubara, and T . Tosa, Appl . Environ . Microbiol . 59:2857-2863, 1993) . In order to increase the productivity of D-biotin, the biotin (bio) operons were cloned from strains SB412, SB304, and 8000 (wild-type strain), and pLGM412, pLGM304, and pLGW101, respectively, were obtained through subcloning . These plasmids harbored 7.2-kb DNA fragments coding for the bioABFCD genes on a low-copy-number vector and were introduced into SB304, SB412, and 8000 . Among the resulting recombinant strains, SB412(pLGM304) exhibited the highest D-biotin production (200 mg/liter) in the production medium . The plasmid was stably maintained in cells . Unexpectedly, SB412(pLGM412) grew very slowly, and the D-biotin productivity of this recombinant strain was not evaluated because pLGM412 was unstable. Anal Biochem, 1993 Oct, 214(1), 212 - 21 Isolation and characterization of the outer membrane proteins of Serratia marcescens W225; Larsen BS et al.; The study addressed the general problem of fractionating cell envelopes in order to isolate the outer membranes of gram-negative bacteria . Whereas the cells are normally transformed into spheroplasts prior to disintegration and membrane separation, Serratia marcescens was found to be resistant to spheroplast formation using the procedures available, which were originally developed for Escherichia coli . An efficient technique for spheroplasting S . marcescens was therefore developed; this comprised combining osmotic shock and lysozyme-EDTA treatment of sucrose-conditioned cells . Spheroplasting efficiency and the amount of outer membrane protein recovered were highly dependent on the spheroplasting technique used . Separation of the outer and inner membranes was performed by two methods, isopyenic centrifugation and selective detergent solubilization with Sarkosyl . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the analysis of specific inner membrane marker enzymes revealed that the protein obtained by detergent solubilization was much purer than that obtained by isopycnic centrifugation . The outer membrane isolated accounted for 60% of the envelope proteins and had a buoyant density of 1.2502 g/cm3 . The protein profile of the outer membrane determined by SDS-PAGE resolved into 12 distinct protein bands, 3 of which represented major proteins. Optom Vis Sci, 1993 Oct, 70(10), 839 - 42 Ultraviolet disinfection of contact lenses; Harris MG et al.; To evaluate the efficacy of ultraviolet (UV) radiation as a method of disinfecting contact lenses and their storage solutions, we contaminated soft lenses (Bausch & Lomb Optima 38), rigid gas permeable (RGP) lenses (Oxyflow F-30), and their storage solutions with three common bacteria . Escherichia coli (E.c.), Staphylococcus epidermis (S.e.), and Serratia marcescens (S.m.) . The storage solutions used were saline solution and RGP conditioning solution . We determined the exposure times to 253.7-nm wavelength UV radiation necessary to disinfect the contact lenses and solutions . The decimal reduction values (D values) found for UV radiation were 10 to 200 hundred times shorter than reported for currently available disinfection systems . For E.c., sterilization was attained after 100 s of exposure . For S.e . and S.m., sterilization occurred after 300 s of exposure . Different contact lens solutions transmit UV radiation to various degrees, with saline solution passing more than 90% of the UV radiation . Thus, our results indicate that UV radiation is an effective and rapid method of disinfecting contact lenses and their storage solutions. Transfusion, 1993 Oct, 33(10), 802 - 8 Transfusion-associated Serratia marcescens infection: studies of the mechanism of action; Gong J et al.; The growth of two strains of Serratia marcescens in blood components was tested in this study . One of the strains had been implicated in the epidemic of transfusion-associated sepsis experienced in Denmark and Sweden in 1991 . In whole blood with a final concentration of 100 colony-forming units per mL of S . marcescens, there was an immediate reduction of more than 95 percent of colony-forming units, but no reduction of the bacterial concentration if the blood had been white cell-reduced before inoculation . This is interpreted as an effect due to phagocytosis by white cells and as a lack of bactericidal effect of the plasma . A reduction to 10 percent of the original concentration, observed if the blood had a nominal content of white cells, was most likely due to phagocytosis . White cell reduction by filtration after inoculation further reduced the bacterial concentration of one of the strains tested, but, after a 1-week lag phase, growth accelerated to high concentrations by 6 weeks . In platelet-rich plasma prepared from S . marcescens-inoculated units, abundant growth was found after 24 hours, increasing to very high concentrations (10(12) colony-forming units/mL) during 10-day storage at 22 +/- 2 degrees C . Keeping the whole blood at ambient temperature for 20 hours before preparation of platelet-rich plasma caused only temporary reduction of bacterial concentration in the S . marcescens experiments, but resulted in a complete absence of bacteria in the platelet-rich plasma for 10 days in control experiments performed with Staphylococcus epidermidis.(ABSTRACT TRUNCATED AT 250 WORDS) Biull Eksp Biol Med, 1993 Oct, 116(10), 384 - 6 {The effect of stimulation of the mononuclear phagocyte system on the binding of high- and low-density lipoproteins by the hepatocytes, Kupffer cells and liver endotheliocytes of rats}; Usynin IF et al.; The role of the mononuclear phagocyte system (MPS) in the regulation of low- (LDL) and high-density lipoprotein (HDL) receptor activity in different rat liver cells was investigated . MPS was activated by intravenous administration of bacterial lipopolysaccharide (LPS) from Serratia marcescens . Liver cells were isolated by in vitro perfusion of the liver with a collagenase solution . Separation of Kupffer and endothelial cells was performed by the method of centrifugal elutriation . In control rats, the hepatocytes bound 1.6 times more 125I-HDL and 2.5 times more 125I-LDL per cell than Kupffer cells . Treatment of rats with LPS resulted in a 4.5-fold decrease in the 125I-HDL binding to Kupffer cells . In contrast, the hepatocytes from LPS-treated rat bound 2 times more 125I-HDL than that from untreated rats . The binding of 125I-LDL and 125I-HDL to endothelial cells and 125I-LDL to hepatocytes were not affected by LPS treatment . These results suggest that the MPS (especially Kupffer cells) plays an important role in the regulation of HDL receptor activity in hepatocytes. Am J Dent, 1993 Oct, 6(5), 239 - 42 The antimicrobial activity of Prevention mouthrinse; Drake DR et al.; Prevention mouthrinse was designed to serve as a vital supplement to normal oral hygiene procedures . To determine the antimicrobial potency of this mouthrinse, minimal inhibitory concentrations (MICs), bactericidal kinetics, and short-term exposure studies were conducted . A spectrum of oral microorganisms was employed in this investigation: Streptococcus sanguis, Streptococcus anginosus, Streptococcus mutans, Actinomyces viscosus, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Eikenella corrodens, Porphyromonas gingivalis, Serratia marcescens and Candida albicans . Microorganisms were cultured in standard enriched media and under appropriate atmospheric conditions . Inhibition assays were conducted in tubes, with each mouthrinse dilution assayed in triplicate . MIC determinations revealed that all of the microorganisms studied were highly susceptible to Prevention mouthrinse, with MICs ranging from 16-fold to 128-fold dilutions . Bactericidal kinetics assays showed rapid killing of the test organisms in the presence of the mouthrinse . Brief (5-minute) exposure of S . mutans to 8-fold diluted mouthrinse resulted in a substantial delay in growth . Under the constraints of this type of study, Prevention mouthrinse exhibited potent antimicrobial activity against all of the microorganisms studied . We support the notion that Prevention mouthrinse may be a valuable supplement to normal oral hygiene procedures . A 6-month clinical trial assessing the in vivo efficacy of Prevention mouthrinse is currently being conducted. FEBS Lett, 1993 Sep 27, 331(1-2), 134 - 40 Astacins, serralysins, snake venom and matrix metalloproteinases exhibit identical zinc-binding environments (HEXXHXXGXXH and Met-turn) and topologies and should be grouped into a common family, the 'metzincins'; Bode W et al.; The X-ray crystal structures of two zinc endopeptidases, astacin from crayfish, and adamalysin II from snake venom, reveal a strong overall topological equivalence and virtually identical extended HEXXHXXGXXH zinc-binding segments, but in addition a methionine-containing turn of similar conformation (the 'Met-turn'), which forms a hydrophobic basis for the zinc ion and the three liganding histidine residues . These two features are also present in a similar arrangement in the matrix metalloproteinases (matrixins) and in the large bacterial Serratia proteinase-like peptidases (serralysins) . We suggest that these four proteinases represent members of distinct subfamilies which can be grouped together in a family, for which we propose the designation, metzincins. Biochim Biophys Acta, 1993 Sep 3, 1202(1), 13 - 21 Characterization of Serratia marcescens nuclease isoforms by plasma desorption mass spectrometry; Pedersen J et al.; Isoforms of Serratia marcescens nuclease found in the natural nuclease produced by S . marcescens and in recombinant nuclease produced by Escherichia coli were structurally characterized by peptide mapping using plasma desorption mass spectrometry . The nuclease isoforms produced and secreted from S . marcescens B10M1, which are present in much greater amounts than in S . marcescens W225 nuclease produced by E . coli, were characterized completely and the information used to facilitate characterization of the recombinant nuclease isoforms . After purification of the nuclease the isoforms were separated on a DEAE-cellulose anion-exchange column and then digested with endoproteinase Lys-C . The peptides generated were isolated by reverse-phase HPLC and their molecular masses determined by plasma desorption mass spectrometry . Comparison of the peptides from the native nuclease, Sm2, and the two isoforms, Sm1 and Sm3, revealed that they differed only in the N-terminus, the latter being found to lack three amino acids in Sm1 and one amino acid in Sm3 . No interior post-translational changes were found in either of the three isoforms . Using this information we were able to confirm that Sm1, the isoform lacking three amino acids, was also present in very small amounts in recombinant S . marcescens W225 nuclease produced and excreted by E . coli. J Clin Oncol, 1993 Sep, 11(9), 1746 - 50 Clinical trial of Serratia marcescens extract and radiation therapy in patients with malignant astrocytoma; Black KL et al.; PURPOSE: A clinical trial was undertaken to determine the safety and efficacy of combining a biologic response modifier derived from the bacterium Serratia marcescens (ImuVert) and radiation therapy (RT) in patients with newly diagnosed anaplastic astrocytoma (AA) or glioblastoma multiforme (GBM) . PATIENTS AND METHODS: Fifteen patients who had undergone either a gross total resection, a partial resection, or a biopsy were treated concurrently with ImuVert and RT . Safety and tolerance were examined by assessment of symptomatic reactions recorded at each ImuVert treatment . Efficacy of treatment was examined in terms of time to progression of tumor and survival . RESULTS: All patients experienced local reactions at the injection sites that consisted of erythema and induration . The majority of patients experienced flu-like symptoms . Hypotension was responsible for the most significant morbidity (which required fluid resuscitation and extended observation) and dose deescalation . No patients were removed from the study because of toxicity . There were no on-study deaths related to ImuVert treatment . Median time to progression was 33.4 weeks, and median survival was 78 weeks . CONCLUSION: These results compare favorably with those of recent studies in patients with malignant astrocytomas who received multimodality therapy. Dev Comp Immunol, 1993 Sep-Oct, 17(5), 377 - 87 Analysis of the attraction of haemocytes from Mytilus edulis by molecules of bacterial origin; Schneeweiss H et al.; Boyden chamber assays were performed to test the stimulatory effect of different bacterial products on the migratory activity of Mytilus haemocytes . The results indicate that these blood cells exhibit chemotactic as well as chemokinetic reactions . Lipopolysaccharides (LPS) from both Serratia marcescens and Escherichia coli stimulated the migration of cells through the membrane of the Boyden chamber when LPS was present in the lower compartment only . In contrast, addition of LPS to the lower as well as to the upper chamber did not increase the rate of migrating cells . Thus, LPS seemed to act as a chemotaxis-stimulating substance . Further analysis indicated that complete LPS molecules are required for cell stimulation because this did not occur when either the lipid or polysaccharide moieties of LPS were tested alone . Unlike LPS, the formylated tripeptide N-FMLP stimulated random cell migration . The peptide, which is released by bacteria, induced a higher haemocyte motility when present in both wells of the Boyden chamber than in tests where it was added to the lower compartment only . This chemokinetic response was not stimulated by the tetrapeptide N-FMLPLys . These findings demonstrate that bacterial products may elicit chemotactic and/or chemokinetic reactions in haemocytes from an invertebrate, and that the type of reaction that occurs is dependent upon the nature of the molecules presented. Indian J Med Res, 1993 Sep, 97, 202 - 5 An outbreak of Serratia marcescens infection among obstetric patients; Stephen M et al.; An outbreak of S . marcescens infection occurred among 17 obstetric patients during May-June 1990 . Simultaneously 11 newborns were also affected . All the 28 strains were identical in their biochemical characteristics, serotype and phage type as well as antimicrobial susceptibility pattern . The source of infection was traced to a contaminated batch of cream, consisting of 0.5 per cent savlon in carboxy methyl cellulose base, used while doing pelvic examination . The affected patients were treated with appropriate antibiotics and there was no mortality . No further infection was reported after the removal of the contaminated cream. Salud Publica Mex, 1993 Sep-Oct, 35(5), 440 - 7 {An epidemic of primary bacteremia due to an endemic strain of Serratia marcescens in an intensive care unit}; Volkow-Fernandez P et al.; An outbreak of Serratia marcescens bacteremia detected in the intensive care unit (ICU) of a tertiary care center on the last days of October, 1985, is described . The rate of primary S . marcescens nosocomial bacteremia during the pre-epidemic period (January-September 1985) was 6.25 per cent; and for the post-epidemic period compared with the epidemic were significantly different (p < 0.0001) . The outbreak strains belonged to the biotype A8b, which has been endemic in our hospital . The responsible organism exhibited an unusual antimicrobial resistance pattern associated to the presence of a specific plasmid (greater than 50 kilobases), which showed similar fragments after restriction endonuclease digestion . No specific risk factors were identified in the case-control study . The outbreak was probably related to a greater influx of infected patients, resulting in less careful infection control measures, due to the emergency situation which suffered the hospital after the earthquakes in 1985 . The unusual high rate of blood isolation of S . marcescens at the ICU was the first sign of the outbreak . The prompt reinforcement of infection control policies facilitated its resolution. Eur J Pediatr, 1993 Sep, 152(9), 745 - 6 Brain abscesses in neonates--report of three cases; Ries M et al.; We report three newborns with brain abscesses . Two infants suffered from Serratia marcescens meningitis and one infant had enterococcal sepsis and meningitis . Brain abscesses were detected by cerebral sonography . Outcome in one infant with S . marcescens infection was poor . This patient developed multicystic encephalo-malacia and severe developmental retardation . In the other patient with S . marcescens infection surgical drainage of the abscess was performed . The outcome was good both in this infant and in the patient with enterococcal brain abscess. J Pharm Belg, 1993 Sep-Oct, 48(5), 341 - 51 {Comparison of in vitro activity of six disinfectants on bacteria from contamination in hemodialysis water}; Ossia-Ongagna Y et al.; The bactericidal and sporicidal activities of six disinfectants were compared in vitro by determination minimal bactericidal concentrations from AFNOR standards for antiseptics and disinfectants . These disinfectants were tested against seventeen bacterial strains and two suspensions of spores including reference strains and wild strains from water of hospital environment . The compound with peracetic acid, ACETOPER 200, is the only disinfectant with the two activities against all strains at 21 degrees C +/- 1 degree C in short length of time . Furthermore except for Serratia liquefaciens that is remarkably resistant to STERLANE, no significant differences of susceptibility to disinfectants appear between wild bacterial strains and reference strains. Mol Microbiol, 1993 Sep, 9(6), 1229 - 37 Amino acid replacements in the Serratia marcescens haemolysin ShIA define sites involved in activation and secretion; Schonherr R et al.; The haemolysin of Serratia marcescens (ShIA) is translocated through the cytoplasmic membrane by the signal peptide-dependent export apparatus . Translocation across the outer membrane (secretion) is mediated by the ShIB protein . Only the secreted form of ShIA is haemolytic . ShIB also converts in vitro inactive ShIA (ShIA*), synthesized in the absence of ShIB, into the haemolytic form (a process termed activation) . To define regions in ShIA involved in both processes, ShIA derivatives were isolated and tested for secretion and activation . Analysis of C-terminally truncated proteins (ShIA) assigned the secretion signal to the amino-terminal 238 residues of ShIA . Trypsin cleavage of a secreted ShIA' derivative yielded a 15 kDa N-terminal fragment, by which a haemolytically inactive ShIA* protein could be activated in vitro . It is suggested that the haemolysin activation site is located in this N-terminal fragment . Replacement of asparagine-69 and asparagine-109 by isoleucine yielded inactive haemolysin derivatives . Both asparagine residues are part of two short sequence motifs, reading Ala-Asn-Pro-Asn, which are critical to both activation and secretion . These point mutants as well as N-terminal deletion derivatives which were not activated by ShIB were activated by adding a non-haemolytic N-terminal fragment synthesized in an ShIB+ strain (complementation) . Apparently the activated N-terminal fragment substituted for the missing activation of the ShIA derivatives and directed them into the erythrocyte membrane, where they formed pores . It is concluded that activation is only required for initiation of pore formation, and that in vivo activation and secretion are tightly coupled processes . Complementation may also indicate that haemolysin oligomers form the pores. J Biol Chem, 1993 Aug 25, 268(24), 17711 - 5 Effect of microbial and mite proteases on low and high molecular weight kininogens . Generation of kinin and inactivation of thiol protease inhibitory activity; Maruo K et al.; Kinin release from guinea pig plasma high molecular weight kininogen (HMWK) induced by various microbial and mite proteases has been demonstrated previously (Molla, A., Yamamoto, T., Akaike, T., Miyoshi, S., and Maeda, H . (1989) J . Biol . Chem . 264, 10589-10594; Maruo, K., Akaike, T., Matsumura, Y., Kohmoto, S., Inada, Y., Ono, T., Arao, T., and Maeda, H . (1991) Biochim . Biophys . Acta 1074, 62-68) . In this paper, we describe the effects of various microbial and mite proteases on low molecular weight kininogen (LMWK) and HMWK from human plasma . A protease from the house dust mite Dermatophagoides farinae (Df-protease) directly liberated kinin from both LMWK and HMWK to a significant degree . The Km, kcat, and kcat/Km values for kinin generation from LMWK were 3.24 microM, 0.61 s-1, and 1.9 x 10(5) M-1 x s-1, respectively, and those for kinin generation from HMWK were 0.56 microM, 0.12 s-1, and 2.1 x 10(5) M-1 x s-1, respectively; kcat/Km values for Df-protease were comparable with that for glandular kallikrein . In contrast, microbial proteases showed only weak kinin-releasing activity from both human plasma kininogens . Four of ten different microbial proteases liberated kinin from LMWK, and only serratial 56-kDa protease released kinin from HMWK . Furthermore, Df-protease markedly inactivated the thiol protease inhibitory activity of LMWK and HMWK, whereas all microbial proteases (as well as the endogenous protease trypsin) did not affect this inhibitory activity of both kininogens from human plasma. J Chromatogr, 1993 Aug 20, 645(2), 353 - 61 Separation of isoforms of Serratia marcescens nuclease by capillary electrophoresis; Pedersen J et al.; Three S . marcescens nuclease isoforms differing mainly in charge (native nuclease with pI 6.8 and two minor isoforms with pI 7.3 and 7.4) were separated using several different modes of high-performance capillary electrophoresis . Separation of the isoforms by free solution capillary electrophoresis was unsatisfactory . Separation by micellar electrokinetic capillary chromatography was therefore investigated in detail and the method optimized with respect to pH and sodium dodecyl sulphate concentration; in addition, the effect of adding various substances to control dispersion and avoid analyte adsorption at the capillary wall was examined . Under optimal conditions there was almost complete baseline separation of the two isoforms with basic pI whereas there was only partial separation of the native form and the isoform with pI 7.4 . With capillary isoelectric focusing there was complete baseline separation of the native nuclease and the other two isoforms. Minerva Chir, 1993 Aug, 48(15-16), 851 - 6 {Is the surgical treatment of spontaneous bacterial peritonitis still up-to-date?}; Aldrighetti L et al.; We describe a case of spontaneous bacterial peritonitis in a 53 year old man affected by cryptogenic micro-macronodular cirrhosis, portal hypertention, splenomegaly and hypersplenism, who was admitted with hepatic failure and septic shock and successfully treated with antibiotics (combination of clindamycin and netilmycin), surgical abdominal drainage and splenectomy . This case gave reason for a literature review and an update on the therapeutic options in these high risk patients, especially concerning the role of surgery . Spontaneous Bacterial Peritonitis (SBP) is defined as a bacterial infection of ascitic fluid in the absence of any septic focus . It is a typical life-threatening complication of hepatic cirrhosis with ascites . Mortality is very high and ranges from 75% to 97% of patients, due to septic shock and hepatic failure (hepatorenal syndrome, hepatic encephalopathy, gastrointestinal bleeding) . Infection with a single organism is found in most cases . Gram negative bacilli are present in about 70% of cases and E . coli (less frequently Klebsiella, Serratia, Pseudomonas) is principally found . Gram positive cocchi comprise an additional 30% of cases . Anaerobic and microaerophilic organisms seem to be rare causes of SBP (2.7-6%); this finding is probably due to the intrinsic bacteriostatic activity of ascites, which contains high oxygen tension (70 mmHg) and is an inhospitable environment for bacteroides and Clostridia . The prevalent isolation of enteric organism suggest that the gut is the most frequent source of infection, even if the pathogenetic mechanism is not yet well known . The right treatment depends on differentiating primary (SBP) from secondary peritonitis.(ABSTRACT TRUNCATED AT 250 WORDS) Epidemiol Infect, 1993 Aug, 111(1), 99 - 107 Comparison of serotype, biotype and bacteriocin type with rDNA RFLP patterns for the type identification of Serratia marcescens; Alonso R et al.; Variations in rDNA gene loci in DNA digests of 209 clinical isolates of Serratia marcescens were determined with an Escherichia coli rRNA probe . Forty-one restriction fragment length polymorphism patterns (ribotypes) were identified, based on the size of 4-14 (mean 7.5) hybridization bands . The patterns differed by more than a single band in 98% of pair-wise comparisons . On a subset of 76 isolates, ribotyping proved to be marginally more discriminating than biotyping (discrimination index 0.92 v . 0.89) followed by serotyping (0.87) and bacteriocin typing (0.74) . About one-third of isolates belonged to unique ribotypes and only two ribotypes exceeded 5% in frequency (23.0 and 6.4% respectively) . A combination of serotype or biotype with ribotyping defined a similar number of strains, although none of the methods alone was sufficiently discriminatory to identify strains . We conclude that due to the accessibility of biotyping and the lack of commercially available antisera for S . marcescens, the biotype and ribotype together provide reliable markers of strain identity. Antibiot Khimioter, 1993 Aug-Sep, 38(8-9), 16 - 21 {Effect of nalidixic acid and mitomycin C on growth and biosynthesis of extracellular proteins by Serratia marcescens}; Iusupova DV et al.; The effect of various concentrations of nalidixic acid and mitomycin C on the dynamics of growth of Serratia marcescens and synthesis of endonuclease and other extracellular enzymes by it was studied . The synthesis of extracellular endonuclease was induced during inhibition of the cell division under the effect of nalidixic acid and mitomycin C . The induction of the endonuclease synthesis preceded the start of the division of the resistant cells which overcame the phase of the population retarded growth . The fermentation broth of the cells exposed to nalidixic acid and mitomycin C contained increased amounts of protein and possessed higher activities of chitin phosphodiesterase and phosphatase . It was shown by PAAG-electrophoresis with sodium dodecyl sulphate that the pattern of the extracellular proteins of S . marcescens undergone quantitative and qualitative changes under the effect of nalidixic acid and mitomycin C. J Appl Physiol, 1993 Jul, 75(1), 233 - 9 Protective effects of E5, an antiendotoxin monoclonal antibody, in the ovine pulmonary circulation; Chen TY et al.; The cross-protective effects of a murine immunoglobulin M monoclonal antilipid A antibody (E5 MAb) were tested by challenging awake sheep with mixtures of in vitro incubated E5 MAb (0.02 mg/kg) with lipopolysaccharide (LPS, 0.02 micrograms/kg) derived from Escherichia coli O111:B4, E . coli O55:B5, or Serratia marcescens . Intravenous infusion of these LPS preparations without antibody into awake sheep produced a similar pattern of fever, leukopenia, plasma thromboxane B2 (TxB2) release, and acute pulmonary vasoconstriction with pulmonary hypertension . The addition of MAb E5 to LPS from E . coli O111:B4 reduced these responses to the LPS in a fashion comparable to that achieved with an MAb specific to the E . coli O111:B4 O-side chain . Incubation of LPS derived from E . coli O55:B5 with the E5 MAb only slightly diminished acute pulmonary hypertension, the delayed temperature increase, and the degree of leukopenia (all P = NS) but reduced the mean peak TxB2 at 60 min (P < 0.05) compared with a control infusion of E . coli O55:B5 LPS . We were unable to demonstrate any protective effects on the pulmonary circulation from incubating E5 with LPS derived from S . marcescens . Preincubation of B55 MAb (a murine immunoglobulin M MAb directed against a human milk fat globulin), the control antibody, with LPS from E . coli 0111:B4 decreased the mean peak TxB2 but had no effect on the other parameters . We conclude that incubating E5 with LPS protects the pulmonary circulation of sheep from challenge with LPS derived from the parent E . coli strain . There were trends toward protection by E5 against LPS from 055:B5 E . coli, but these did not reach statistical significance.(ABSTRACT TRUNCATED AT 250 WORDS) Antimicrob Agents Chemother, 1993 Jul, 37(7), 1456 - 62 Characterization of the aac(6')-Ib gene encoding an aminoglycoside 6'-N-acetyltransferase in Pseudomonas aeruginosa BM2656; Galimand M et al.; Pseudomonas aeruginosa BM2656 was resistant to tobramycin and susceptible to gentamicin and amikacin by disk diffusion testing . This unusual resistance was not transferable by conjugation to Escherichia coli or P . aeruginosa PAO38, and plasmid DNA was not detected in this strain . A 0.9-kb fragment harboring the tobramycin resistance gene was cloned from BM2656 into pUC18, generating pAT129 . Analysis for aminoglycoside-modifying activity in extracts of BM2656 and E . coli harboring pAT129 indicated that tobramycin resistance was due to synthesis of an aminoglycoside 6'-N-acetyltransferase type I {AAC(6')-I} enzyme which modified amikacin and tobramycin . Although amikacin was acetylated, the bactericidal synergism of this aminoglycoside with ceftazidime against BM2656 was minimally affected . The sequence of the DNA fragment was determined . It contained an aac (6')-Ib-like gene and was located downstream from a conserved region related to Tn21 . The translated sequence of this aac(6')-Ib gene possessed 99.2% identity with the putative products of the aac(6')-Ib gene cassettes from Serratia marcescens and Klebsiella pneumoniae and 69% identity with the putative aacA(6')-II gene product from P . aeruginosa . We conclude that an aac(6')-Ib gene has spread to the chromosome of P . aeruginosa, probably by transposition. J Clin Microbiol, 1993 Jul, 31(7), 1826 - 30 Growth and survival of Serratia marcescens under aerobic and anaerobic conditions in the presence of materials from blood bags; Szewzyk U et al.; Several patients receiving blood transfusions during the summer of 1991 developed bacteremia after the transfusion . In all cases, the infection was caused by Serratia marcescens . The same strain of Serratia marcescens was isolated from the patients and from the outer surface of unfilled blood bags . The transport containers for the blood bags were made anoxic by using a catalyst in order to prevent microbial growth . The survival and growth of S . marcescens K202, which was isolated from the blood bags, was studied at different oxygen concentrations in deionized water containing materials derived from the blood bags . The rate of survival and growth of S . marcescens was highest under anaerobic conditions, in which growth occurred with all materials and even in deionized water alone . In contrast, S . marcescens did not survive in control cultures under semi-anaerobic and aerobic conditions . Growth was observed, however, under both aerobic and semi-anaerobic conditions in the presence of each of the tested blood bag materials . These findings indicate that the conditions in the transport containers for the blood bags were favorable for the survival and growth of S . marcescens. Biochim Biophys Acta, 1993 Jun 11, 1157(2), 178 - 84 Purification and characterization of the valine sensitive acetolactate synthase from Serratia marcescens ATCC 25419; Yang JH et al.; The valine sensitive acetolactate synthase (ALS) isozyme from Serratia marcescens ATCC 25419 was purified to homogeneity . Analysis of the native molecular weight of the purified enzyme by the native pore gradient polyacrylamide gel electrophoresis indicated the molecular weight of about 178,000 and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the enzyme to be composed of two different types of subunits with molecular weights of 62,000 and 35,000 . The molar ratio of the two polypeptides was estimated to be 1, suggesting that native enzyme is composed of two large subunits and two small subunits . The enzyme exhibits homotropic allosterism with pyruvate unlike other enteric ALS isozymes . The specificity ratio R (V{acetohydroxybutyrate}/V{acetolactate} = R.{alpha-ketobutyrate}/pyruvate}), of the enzyme was found to be 0 suggesting that the Serratia ALS has very high specificity for pyruvate . The pH optimum was around 7.5, and the enzyme was stable at 50 degrees C for 30 min . The pI value for the purified enzyme was 5.2 . The concentration of branched chain amino acids for 50% inhibition of the enzyme was 0.1 mM for valine, and 1 mM for leucine and isoleucine, respectively. Dtsch Med Wochenschr, 1993 Jun 11, 118(23), 854 - 60 {Whipple's disease associated with opportunistic infections}; Meier-Willersen HJ et al.; A 36-year-old man, with a history of recurrent respiratory infection, dermatomycosis, arthralgia and abnormal stools for 12 years, developed a febrile illness (up to 40 degrees C) . A Serratia marcescens septicaemia responded to antibiotics . Four months later cervical and abdominal lymph-adenopathies were noticed . Cervical lymph node biopsy revealed lymphadenitis with epithelioid cell nests . Duodenoscopy with biopsy demonstrated Whipple's disease associated with lambliasis . Electron-microscopy showed rod-shaped bacteria typical of Whipple's disease, and Giardia lamblia . Using the polymerase chain reaction, Whipple-specific DNA fragments of 284 base pairs from the genome of the Whipple bacterium (Tropheryma whippelii) were demonstrated . Antibiotic treatment with Ampicillin (2 g three times daily) and ceftriaxone (2 g once daily) i.v . for 21 days, followed by oral ofloxacin (200 mg daily) and co-trimoxazole (three times daily 800 mg sulfamethoxazole and 160 mg trimethoprim), brought about remission of Whipple's disease . Long-term antibiotic treatment was continued with co-trimoxazole . Lambliasis recurred after 3 and 5 months, despite treatment with metronidazole, 250 mg three times daily for 7 days. Ann Plast Surg, 1993 Jun, 30(6), 556 - 9 Tourniquet-related hypotension in venous stasis ulcer excision; Feldman DL et al.; Extremity tourniquets are widely used to achieve bloodless dissection in the surgical field . Excision of venous stasis ulcers (VSU) is aided by tourniquet use because of large dilated veins associated with venous stasis disease . We present 3 patients with hypotensive shock occurring 10 to 15 minutes after tourniquet release after excision of venous stasis ulcers . All patients had long histories of venous stasis changes and two-thirds had prior histories of deep vein thromboses and pulmonary embolism . Mean tourniquet inflation time was 34 minutes and there were electrocardiographic changes in two-third of the patients . All patients responded rapidly to standard resuscitation measures and in all 3 postoperative testing for pulmonary embolus and myocardial infarction was negative . Wound cultures revealed no organisms in 1 patient, mixed Gram-positive cocci in another, and greater than 10(5) Serratia marcescens in the third patient . Although small decreases in blood pressure and blood pH, and increases in blood lactate, PcO2, and creatinine phosphokinase, are normally associated with the use of extremity tourniquets, hypotensive shock has not been reported . The combined effect of tourniquet ischemia and venous stasis changes may cause hypotensive shock by (1) an endotoxic bolus upon tourniquet release, (2) pulmonary microembolization of platelet, fibrin, and leukocyte aggregates causing vasoactive substance release, and (3) synergistic effects of platelet-activating factor, a known mediator of endotoxic shock . The untoward events noted in these patients may be prevented by (1) proximal to distal dissection of the ulcer with initial ligation of large veins, (2) pretreatment with steroids and/or platelet-activating factor antagonists, and/or (3) slow release of the tourniquet. J Antimicrob Chemother, 1993 Jun, 31(6), 841 - 54 The use of a DNA probe and PCR to examine the distribution of the aac(6')-Ic gene in Serratia marcescens and other gram-negative bacteria; Snelling AM et al.; A nucleotide probe for the chromosomal aminoglycoside 6' acetyltransferase gene (aac(6')-Ic) of Serratia marcescens was used in non-radioactive dot-blot hybridization experiments with 186 strains belonging to ten different species of Serratia . The gene was only detected in strains of S . marcescens (all strains tested), and positive hybridization was seen irrespective of whether or not strains were kanamycin-resistant . An additional 180 strains belonging to 28 Gram-negative bacterial species other than in the genus Serratia did not hybridize with the probe . A rapid PCR test for the aac(6')-Ic gene was developed and used to confirm that the aac(6')-Ic gene is only found in S . marcescens . Southern hybridization analysis of S . marcescens chromosomal DNA demonstrated that the gene was usually located on a PvuII fragment with a putative isoleucine tRNA-2 gene, but polymorphisms with respect to the size of this fragment were observed. Biol Chem Hoppe Seyler, 1993 Jun, 374(6), 363 - 76 Microbial metabolism of quinoline and related compounds . XVIII . Purification and some properties of the molybdenum- and iron-containing quinaldic acid 4-oxidoreductase from Serratia marcescens 2CC-1; Fetzner S et al.; Serratia marcecens 2CC-1 utilizes quinaldic acid (quinoline 2-carboxylic acid) as sole source of carbon, nitrogen and energy . Growth of strain 2CC-1 on quinaldic acid as well as on nicotinic acid and hypoxanthine was inhibited completely by the molybdate antagonist tungstate, whereas growth on kynurenic acid and 6-hydroxynicotinic acid was not affected by tungstate . The synthesis of the molybdenum-containing hydroxylases quinaldic acid 4-oxidoreductase and nicotinic acid 6-oxidoreductase was found to be inducible . In addition, Serratia marcescens 2CC-1 produced a constitutively expressed xanthine oxidoreductase . Quinaldic acid 4-oxidoreductase was purified 1075-fold with a recovery of 5% . For catalytic activity, artificial electron acceptors were necessary . The 95-100-kDa enzyme was a heterodimer with subunit molecular masses of 75-80 kDa and 18-19 kDa . Quinaldic acid 4-oxidoreductase contained 2.3-3.7 g atom of iron and 0.5-0.6 g atom of molybdenum per mol of enzyme . The absorption spectrum exhibited maxima at 280 nm, 334 nm, 480 nm and a shoulder at 550 nm, with A280/A334 = 4.8, A280/A450 = 10.0, A280/A480 = 9.4, and A450/A550 = 1.6, suggesting the absence of a flavin cofactor . Acridine, quinacrine, ethylenediaminetetraacetate, 2,2'-dipyridyl, 1,10-phenanthroline and iodoacetate did not affect enzyme activity . p-Hydroxymercuribenzoate, m-arsenite, cyanide and methanol were effective inhibitors of quinaldic acid 4-oxidoreductase . Cyanide-inhibited enzyme was reactivated by treatment with S2-, indicating the presence of a pterin molybdenum cofactor with a monooxo-monosulfidotype molybdenum center . Quinaldic acid 4-oxidoreductase showed a very high substrate specificity, quinaldic acid being the only substrate found to be transformed significantly. Masui, 1993 Jun, 42(6), 931 - 2 {Septic attack during percutaneous nephrolithotripsy under epidural anesthesia}; Nakata F et al.; A 59-year-old man was scheduled for percutaneous nephrolithotripsy (PNL) for renal calculus . Anesthesia was maintained by epidurally administered mepivacaine 1.5% under prone position . During surgical manipulation, the patient complained of shivering and chill accompanied with fever and tachycardia . His temperature peaked at 40.7 degrees C and decreased gradually by cooling . Serratia was detected from arterial blood obtained in the operating room . It was thought that fever attack was due to excessive increase of intrarenal pressure by irrigation fluid which might have led to septicemia . Therefore, careful attention for septic attack must be paid during PNL procedure as well as postoperatively. Gene, 1993 May 15, 127(1), 39 - 45 Cloning and sequencing of the Serratia marcescens gene encoding a single-stranded DNA-binding protein (SSB) and its promoter region; de Vries J et al.; The gene (ssb) coding for a single-stranded DNA-binding protein (SSB) was identified on a 1.2-kb EcoRI-SalI fragment cloned from chromosomal DNA of Serratia marcescens . The cloned fragment conferred increased resistance against UV and mitomycin C (MC) to ssb- mutants of Escherichia coli . The nucleotide (nt) sequence revealed that SSB consists of 175 amino acids (aa) and has an M(r) of 18,677 . It shows 89% aa sequence homology with the SSB of E . coli . The nt sequence preceding the gene contains three promoters . Two of them overlap with a presumptive SOS box, and the distal one overlaps with a second SOS box that coincides with the promoter of the adjacent uvrA (gene encoding the UvrA protein) . The uvrA is transcribed in a direction opposite to that of ssb . The sequence coding for the N terminus of the UvrA of S . marcescens indicates that the first 74 aa are identical to those of the E . coli protein . The results suggest that the two bacterial SSBs are members of a group which differs from the known SSBs of prokaryotic transmissible plasmids, because their aa sequence homology with these proteins is only about 60%. South Med J, 1993 May, 86(5), 489 - 96 Chemotherapy-induced alopecia: new developments; Hussein AM; Alopecia (hair loss) is one of the most physically and psychologically distressing side effects of cancer chemotherapeutic drugs . Since its first recognition as a common outcome to most chemotherapeutic agents, only a few trials have been reported, using either a method to temporarily reduce the scalp blood flow (scalp tourniquet or hypothermia) or vitamin E, with undocumented and variable efficacy . The lack of progress in the treatment and prevention of chemotherapy-induced alopecia is in part due to the lack of a reproducible animal model . In the past 2 years, we reported on the following observations: (1) treatment of 8-day-old rats with vidarabine (ara-C), doxorubicin, and cyclophosphamide consistently produced either total body alopecia (ara-C and cyclophosphamide) or alopecia confined to the head and proximal part of the back (doxorubicin); (2) Imuvert, a biologic response modifier derived from the bacterium Serratia marcescens, uniformly produced complete protection against alopecia induced by ara-C and doxorubicin but not that produced by cyclophosphamide; (3) the protective effect of Imuvert against chemotherapy-induced alopecia is mediated by a monocyte-mediated cytokine; and (4) this monocyte-derived cytokine is, possibly, interleukin-1 . These observations constitute important progress in the understanding and prevention of chemotherapy-induced alopecia. J Lab Clin Med, 1993 May, 121(5), 662 - 7 Changes in cytoplasmic pH in platelets activated through the high- and moderate-affinity receptor pathways by highly purified human alpha-thrombin; Jones GD et al.; Platelets in plasma were loaded with the probe BCECF/AM, and changes in cytoplasmic pH levels induced by highly purified human alpha-thrombin (2900 NIH U/mg) were studied in washed platelets having high- and moderate-affinity receptors and in platelets from which the high-affinity alpha-thrombin receptor had been removed by treatment with Serratia marcescens protease . In intact platelets, cytoplasmic acidification reached a maximum within 2 minutes of -0.072 +/- 0.009 pH units at 0.3 nmol/L alpha-thrombin concentration (0.03 U/ml) . Cytoplasmic pH values were higher at both lower and higher alpha-thrombin concentrations and were significantly (p = 0.018) higher at 2 nmol/L alpha-thrombin, which induced -0.037 +/- 0.013 pH units of acidification . Five nanomoles of alpha-thrombin, however, induced cytoplasmic alkalinization of +0.027 +/- 0.033 pH units . In platelets lacking the high-affinity receptor where there is a 10 to 20-fold reduction in sensitivity to alpha-thrombin, acidification reached a maximum of -0.175 +/- 0.033 pH units at 2 nmol/L alpha-thrombin, but alkalinization was observed at 5 nmol/L (+0.038 +/- 0.025) and 10 nmol/L (+0.042 +/- 0.007) alpha-thrombin . These results show that the transition from acidification to alkalinization occurs in the same range of alpha-thrombin concentrations (2 to 5 nmol/L) in both preparations, despite the rightward shift in sensitivity caused by the absence of the high-affinity receptor . However, the maximum acidification reached in control platelets (-0.037 pH units at 2 nmol/L) was much less than the value obtained in platelets lacking the high-affinity receptor (-0.175 pH units at 2 nmol/L alpha-thrombin).(ABSTRACT TRUNCATED AT 250 WORDS) Ann Thorac Surg, 1993 May, 55(5), 1192 - 6 Coronary artery bypass grafting in patients with chronic lymphocytic leukemia; Finck SJ et al.; Chronic lymphocytic leukemia is a disease of the elderly . It tends to have a variable clinical course . Because of the patients' immunologically dysfunctional state, there has been reluctance to perform open cardiac procedures because of concern about early postoperative sepsis leading to death . To assess the risk of coronary artery bypass grafting in elderly patients, the records of 26 patients (mean age, 69.6 +/- 4.9 years) with chronic lymphocytic leukemia who underwent coronary artery bypass grafting between January 1975 and July 1990 were retrospectively reviewed . Nineteen underwent isolated coronary artery bypass grafting, and 7 had combined procedures . The operative mortality rate was 7.7% . Postoperative infections developed in 6 patients (23.1%): pneumonia in 3 and sternal osteomyelitis, acute parotiditis, and bacteremia in 1 each . One of these 6 patients died of acute Serratia pneumonitis . Twenty-four patients (92.3%) were discharged from the hospital an average of 10.6 +/- 7.7 days postoperatively . Patients with chronic lymphocytic leukemia can undergo coronary artery bypass grafting with acceptable mortality but with increased risk of postoperative infection. FEMS Immunol Med Microbiol, 1993 Apr, 6(4), 265 - 71 Stimulation and inhibition of polymorphonuclear leukocytes phagocytosis by lipoamino acids isolated from Serratia marcescens; Miyazaki Y et al.; The role of the lipoamino acids (serratamolide and ornithine lipid), membrane lipid components of Serratia marcescens, was examined in phagocytosis and phagosome-lysosome fusion of human peripheral polymorphonuclear leukocytes . A mutant strain of Serratia marcescens (NS 38-09) lacking serratamolide was actively phagocytosed by human PMN, while the wild-type strain (NS 38) producing serratamolide was more resistant to phagocytosis by human PMN . Phagocytosis of killed Staphylococcus aureus coated with lipoamino acid (serratamolide), showed that they were more resistant to phagocytosis by PMN, while the cells coated with ornithine lipid or serratamic acid were phagocytosed more actively . Staphylococci coated with phosphatidylethanolamine or phosphatidylglycerol had no significant effect on phagocytosis by PMN . Phagosome-lysosome fusion by PMN labelled with acridine orange was examined by fluorescence microscopy . The fusion indices of lipoamino acid-coated staphylococci were the same as that of controls . Further, ornithine lipid-coated staphylococci stimulated the release of superoxide anion from PMN slightly, but serratamolide did not . These results suggested that serratamolide may contribute to the virulence of S . marcescens in vitro. Antiviral Res, 1993 Apr, 20(4), 279 - 92 Murine cytomegalovirus-inhibitory effects of ImuVert; Sidwell RW et al.; ImuVert, a sterile preparation composed primarily of Serratia marcescens membrane vesicles and ribosomes, was significantly inhibitory to murine cytomegalovirus (MCMV) infections in BALB/c mice . Antiviral activity was manifested as increased survivor number and decreased recoverable virus titers in spleens, lungs and salivary glands . Treatments were intraperitoneal (i.p.) beginning 24 h pre, 4 h post- or 24 h post-virus inoculation and then repeated 4 days later . Doses of 5, 16 or 50 micrograms/mouse were effective; 160 micrograms/mouse, which caused host weight loss in toxicity controls, was not inhibitory to the infection . A single i.p . treatment of mice substantially augmented natural killer (NK) cell activity and increased total B-cells, while reducing total T- and T-helper cells . A late (48 h) decline in T-cell function and transient increases in B-cell function were observed in the treated animals . Serum interferon was not induced . Mice pretreated with anti-asialo GM1 antibody to reduce their NK cell populations, then infected with MCMV and treated with ImuVert were protected to the same degree as normal animals . Severe combined immunodeficient mice infected with MCMV and treated with ImuVert were not protected from the infection . These data suggest ImuVert to act by a mechanism other than NK cell activation in preventing MCMV infections. Mol Microbiol, 1993 Apr, 8(2), 409 - 23 Activity domains of the TonB protein; Traub I et al.; Escherichia coli and related Gram-negative bacteria contain an energy-coupled transport system through the outer membrane which consists of the proteins TonB, ExbB, ExbD anchored in the cytoplasmic membrane and receptors in the outer membrane . Differences in the activities of the Escherichia coli and the Serratia marcescens TonB proteins were used to identify TonB functional domains . In E.coli TonB segments were replaced by equivalent fragments of S . marcescens TonB and the activities of the resulting chimaeric proteins were determined . In addition, E . coli TonB was truncated at the C-terminal end, and point mutants were generated using bisulphite . From the results obtained we draw the following conclusions: an important site of interaction between TonB and ExbB is located in the N-terminal region of TonB within or close to the cytoplasmic membrane since an N-terminal 44-residue fragment of TonB was stabilized by ExbB and interfered with wild-type TonB activity . In addition, the activity of a TonB derivative in which histidine residue 20 was replaced by arginine was strongly reduced, and a double mutant containing arginine-7 to histidine and alanine-22 to threonine substitutions displayed an impaired uptake of ferrichrome . Furthermore, the domain around residue 160 is involved in TonB activity . S . marcescens TonB segments of this region in E . coli TonB conferred S . marcescens TonB activities, and E . coli TonB point mutants displayed strongly impaired activities for the uptake of colicin B and M and ferric siderophores . Plasmid-encoded tonB mutants of this region showed negative complementation of chromosomal wild-type tonB, and certain tonB mutants suppressed colicin B TonB-box mutants . Uptake of colicins required different domains in TonB, for colicin B and M around residue 160 and for colicin Ia, a domain closer to the C-terminal end . Tandem duplication of the E . coli (EP)X(KP) region by insertion of the S . marcescens (EP)X(KP) region (38 residues) and replacement of lysine residue 91 by glutamate did not alter TonB activity so that no evidence was obtained for this region to be implicated in receptor binding . The aberrant electrophoretic mobility of TonB was caused by the proline-rich sequence since its removal resulted in a normal mobility. Mol Microbiol, 1993 Apr, 8(2), 229 - 42 Secretion of Serratia liquefaciens phospholipase from Escherichia coli; Givskov M et al.; The Serratia liquefaciens phospholipase (PhlA) is secreted to the medium from its natural host . Here we present results which indicate that, when cloned and expressed in Escherichia coli, secretion can be mediated by a putative host-encoded pathway, expression of which is controlled by FlhD (formerly FlbB), the master regulator of the flagellar/chemotaxis regulon . In the absence of this secretion pathway, the synthesized phospholipase accumulates inside the host cell where it forms a complex with the PhlB protein . PhlB, which is encoded from the promoter distal gene of the phospholipase operon, inhibits the phospholipase activity of PhlA . Formation of this enzymatically inactive PhlA/PhlB complex is required for maintenance of cell viability. J Hosp Infect, 1993 Apr, 23(4), 263 - 70 An outbreak of Serratia marcescens traced to a contaminated bronchoscope; Vandenbroucke-Grauls CM et al.; An outbreak of colonization and infection with Serratia marcescens in a surgical Intensive Care Unit is described . A case-control study pointed to a bronchoscope as the source of the epidemic strain, and cultures of washing effluent of the incriminated bronchoscope yielded S . marcescens . Discontinuation of the use of the instrument and the implementation of recommendations for future use of bronchoscopes ended the outbreak. FEMS Microbiol Lett, 1993 Apr 1, 108(2), 225 - 30 Antibacterial activity of phosphono peptides based on 4-amino-4-phosphonobutyric acid; Zboinska E et al.; Phosphono dipeptides based on 4-amino-4-phosphonobutyric acid (phosphonic acid analogue of glutamic acid, GluP) were synthesized and evaluated for their antibacterial activity . Dipeptides containing N-terminal alanine, leucine, isoleucine, phenylalanine or lysine showed marked antibacterial activity against Escherichia coli, whilst those containing alanine, leucine, valine or proline were active against Serratia marcescens . AlaGluP and LeuGluP were nearly equipotent with the respective dipeptides based on 1-aminoethylphosphonic acid (phosphonic acid analogue of alanine) . The structure-activity relationship, i.e . dependence of the activity of phosphono dipeptides on the nature of their N-terminal component, indicated that transport of the peptide through the bacterial cytoplasmic membrane constitutes a crucial step in its antibacterial activity. Transfusion, 1993 Mar, 33(3), 221 - 7 Nosocomial epidemic of Serratia marcescens septicemia ascribed to contaminated blood transfusion bags; Heltberg O et al.; Two cases of transfusion-related Serratia marcescens bacteremia prompted extensive epidemiologic investigations in three independent hospitals . Test tubes and plasma from donors whose blood was drawn into bags from a single production batch were cultured . Analysis of the ribotype of S . marcescens isolates was performed . For comparison, a strain from the production plant and eight other, unrelated bacteremia isolates were examined . In addition, a retrospective national survey was carried out . S . marcescens was cultured from 11 (0.73%) of 1515 blood units, and an additional (third) bacteremic patient was identified . The clinical isolates from three patients, the three units of blood transfused, and the plant-derived strain shared a unique ribotype . The incident is interpreted as a sporadic, bacterial contamination of blood bags with the S . marcescens epidemic strain, occurring during the manufacturing or packaging . A similar incident has not previously been reported . Attention is drawn to the possibility of significant contamination during the complex production of multiple-bag blood collection systems . Guidelines for improved registration and handling of transfusion complications in wards are suggested . Manufacturers should be encouraged to provide blood packs with sterile exteriors, in appropriate, single, outer packages. J Foot Ankle Surg, 1993 Mar-Apr, 32(2), 227 - 31 Serratia marcescens as a cause of postoperative infection in a total joint implant; Brink D et al.; Serratia marcescens is a rare cause of musculoskeletal infections and osteomyelitis occurring most frequently in nosocomial infections, debilitated patients, drug addicts, and traumatic open wounds . It is rarely a cause of postoperative infection in elective surgery with only a few such cases being reported in the United States and European literature . The authors present a case of Serratia marcescens infection of a total joint implant from an apparent perioperative source and review the literature concerning this organism as a cause of infection in man. J Mol Biol, 1993 Feb 5, 229(3), 656 - 70 Co-existing structures of an mRNA stability determinant . The 5' region of the Escherichia coli and Serratia marcescens ompA mRNA; Rosenbaum V et al.; The structure of untranslated regions of mRNA is thought to play a key role in the degradation of mRNAs by specific RNases . As a model system, in vitro transcripts of the stability determining 5' non-coding region of bacterial ompA mRNA were investigated by calculation of secondary structure models and by experiments applying the temperature-gradient gel electrophoresis (TGGE) . For the theoretical prediction of secondary structures an algorithm was used, which yields the structure of lowest free energy as well as a large set of suboptimal structures . Three structures were predicted to co-exist in similar concentrations under native conditions . They denature in a low temperature transition leading to a unique structure which denatures in a high temperature transition . The prediction of three structures and two transitions could be confirmed experimentally by TGGE . Due to the use of transcripts of different length the conformational transitions could be attributed to distinct parts of the molecules . A pseudoknot structural motif was predicted theoretically, but could not be confirmed experimentally . Comparing ompA transcripts of E . coli and S . marcescens, a conservation of structural features could be shown in spite of a sequence homology of only 63% . Regarding the sequential folding of the transcript after synthesis, a metastable structure is formed first and is converted slowly into structures of lower free energy . The biological implications for in vivo degradation are discussed. Acta Paediatr Jpn, 1993 Feb, 35(1), 17 - 21 The in vivo and in vitro postantibiotic effect of aminoglycosides using a clinically isolated micro-organism; Arai S et al.; We reported a case of abscess in which Serratia marcescens was isolated as the causative organism . We measured the postantibiotic effect (PAE) of dibekacin (DKB) and gentamicin (GM) against S . marcescens and studied the relationship between the clinical effect and the PAE . The minimal inhibitory concentration (MIC) of DKB against S . marcescens was 6.25 micrograms/mL and the serum concentration 30 min after infusion of 100 mg DKB was 5.99 micrograms/mL . The abscess was cured by the administration of DKB every 12 h . The PAE in vivo was 2.5, 2.9 and 3.3 h when DKB was administered at 50 mg/kg, 100 mg/kg and 200 mg/kg, respectively . This PAE is one of the reasons that infection can be effectively treated with intermittent administration, even if the serum concentration is below the MIC. Appl Environ Microbiol, 1993 Feb, 59(2), 620 - 2 Purification and properties of a thermostable chitinase from Streptomyces thermoviolaceus OPC-520; Tsujibo H et al.; A chitinase was purified from the culture filtrate of Streptomyces thermoviolaceus OPC-520 . The enzyme showed a high optimum temperature (70 to 80 degrees C), a high optimum pH level (8.0 to 10.0), and heat stability . This enzyme showed high sequence homology with chitinases from Serratia marcescens QMB1466 and Bacillus circulans WL-12. J Clin Microbiol, 1993 Feb, 31(2), 444 - 5 Fatal pneumonia due to Serratia proteamaculans subsp . quinovora; Bollet C et al.; Serratia proteamaculans subsp . quinovora was isolated from several samples (blood cultures, tracheal aspirates, pleural effusion) from a patient with pneumonia . This is the first clinical isolate and the first documented human infection caused by this organism. J Bacteriol, 1993 Feb, 175(4), 959 - 65 Role of serine 352 in the allosteric response of Serratia marcescens aspartokinase I-homoserine dehydrogenase I analyzed by using site-directed mutagenesis; Omori K et al.; Aspartokinase I and homoserine dehydrogenase I (AKI-HDI) from Serratia marcescens Sr41 are encoded by the thrA gene as a single polypeptide chain . Previously, a single amino acid substitution of Ser-352 with Phe was shown to produce an AKI-HDI enzyme that is not subject to threonine-mediated feedback inhibition . To determine the role of Ser-352 in the allosteric response, the thrA gene was modified by using site-directed mutagenesis so that Ser-352 of the wild-type AKI-HDI was replaced by Ala, Arg, Asn, Gln, Glu, His, Leu, Met, Pro, Thr, Trp, Tyr, or Val . The Thr-352 and Pro-352 replacements rendered AKIs sensitive to threonine . The Tyr-352 and Asn-352 substitutions led to activation, rather than inhibition, of AKI by threonine . The other replacements conferred threonine insensitivity on AKI . The threonine sensitivity of HDI was also changed by the amino acid substitutions at Ser-352 . The HDI carried by the Tyr-352 mutant AKI-HDI was activated by threonine . Single amino acid replacements at Ser-352 by Ala, Asn, Gln, His, Phe, Pro, Thr, or Tyr were introduced into truncated AKI-HDIs containing the AKI and the central regions . The AKI activity of the truncated AKI-HDI containing the first 468 amino acid residues was sensitive to threonine, and introduction of the amino acid replacements did not alter the threonine sensitivity of the AKI . Another truncated AKI-HDI containing the first 462 amino acid residues possessed threonine-resistant AKI, whereas the substitutions of Ser-352 with Ala and Pro rendered AKI sensitive to threonine . The replacement of GIn-351 with Phe activated AK1 of the truncated AKI-HDI in the presence of L-threonine . These findings suggest that Ser-352 of the central region of AKI-HDI is possibly a key residue involved with the allosteric regulation of both AKI and HDI activities. J Bacteriol, 1993 Feb, 175(3), 785 - 94 Nucleotide sequence of the Serratia marcescens threonine operon and analysis of the threonine operon mutations which alter feedback inhibition of both aspartokinase I and homoserine dehydrogenase I; Omori K et al.; The nucleotide sequence of the Serratia marcescens threonine operon (thrA1A2BC) was determined . Three long open reading frames were identified; these open reading frames code for aspartokinase I (AKI)-homoserine dehydrogenase I (HDI), homoserine kinase, and threonine synthase, in that order . The predicted amino acid sequences of these enzymes were similar to the amino acid sequences of the corresponding enzymes in Escherichia coli . The AKI-HDI protein is apparently a tetramer composed of monomer polypeptides that are 819 amino acids long . A deletion analysis revealed that the central and C-terminal region was responsible for threonine-resistant HDI activity, a monomeric fragment extending from the N terminus to residue 306 was responsible for threonine-resistant AKI activity, and an N-terminal portion containing 468 residues was responsible for threonine-sensitive AKI activity . The thrA(1)1A(2)1 and thrA(1)5A(2)5 mutations of threonine-excreting strains HNr21 and TLr156, which result in the loss of threonine-mediated feedback inhibition of both AKI activity and HDI activity, cause single amino acid substitutions (Gly to Asp at position 330 and Ser to Phe at position 352, respectively) in the central region of the AKI-HDI protein . The thrA1+A(2)2 mutation of strain HNr59, which results in a threonine-sensitive AKI and a threonine-resistant HDI, also causes a single amino acid substitution (Ala to Thr at position 479). Lett Appl Microbiol, 1993 Feb, 16(2), 77 - 9 Rapid extraction of high purity chromosomal DNA from Serratia marcescens; Alonso R et al.; Rapid non-specific degradation of Serratia marcescens DNA extracted with guanidium thiocyanate, occurred within 10 min of incubation with restriction endonuclease enzymes . The described modified method based on chemical and enzymatic deproteinization produced preparations of Ser . marcescens DNA of high yield and quality which did not autodegrade when incubated with restriction endonucleases. Biochim Biophys Acta, 1993 Jan 22, 1180(3), 267 - 76 Difference between human and guinea pig Hageman factors in activation by bacterial proteinases: cleavage site shift due to local amino acid substitutions may determine the activation efficiency of serine proteinase zymogens; Semba U et al.; Human and guinea pig Hageman factors have been subjected to the action of pseudomonal elastase and serratial E15 proteinase . The pseudomonal elastase cleaved 22-24% of the human molecule at Arg353-Val354, and the remainder at Gly357-Leu358 resulting in the generation of about 20% of potential activity as activated Hageman factor, compared with trypsin activation, while it hydrolyzed Arg340-Ile341 bond in guinea pig molecule and generated about 75% of activity as activated Hageman factor . The serratial proteinase did not hydrolyze the essential cleavage site (Arg353-Val354) of the human zymogen but Gly356-Gly357 (30%) and Gly357-Leu358 (70%) bonds . Both products showed no activity . The guinea pig zymogen, in contrast, was cleaved mostly at Arg340-Ile341 (70%) and less abundantly at Gly344-Leu345 (30%), generating about 85% of the whole potential activity as activated Hageman factor . From the high correspondence between the proportions of activation and of hydrolysis at the essential cleavage site in activation, it was concluded that hydrolysis of the bonds different from the essential bond did not cause activation, even when the spatial separation was only 3 or 4 residues . Considering the amino acid differences between human and guinea pig Hageman factors, -Met351-Thr-Arg-Val-Val-Gly-Gly-Leu-Val-Ala360- and -Leu338-Ser-Arg-Ile-Val-Gly-Gly-Leu-Val-Ala347-, respectively, it was realized that even the minor amino acid substitutions caused the cleavage site shift which resulted in significant differences in activation efficiency of the proteinase zymogens. J Biomater Sci Polym Ed, 1993, 4(3), 199 - 215 Mechanism of cytoplasmic calcium changes in platelets in contact with polystyrene and poly(acrylamide-co-methacrylic acid) surfaces; Yui N et al.; Changes in cytoplasmic free calcium levels ({Ca2+}i) in platelets in contact with polystyrene (PSt) and poly(acrylamide-co-methacrylic acid) (PAAmMAc) particles were evaluated and results were compared with those from two representative biological calcium agonists; thrombin and calcium ionophore A23187 . PSt particles stimulated a steep increase in cytoplasmic calcium levels in platelets as much as thrombin and A23187 . Serratia protease-treated platelets showed a steep increase in {Ca2+}i by PSt particles, suggesting that PSt surfaces can initiate platelet activation independent of a glycoprotein Ib (GPIb)-mediated pathway . By contrast, dibucaine-treated platelets showed little increase in {Ca2+}i by PSt particles, indicating that microfilament assembly, including binding of GPIb with actin binding protein, should be required for platelet activation in contact with PSt surfaces . PAAmMAc particles induced little increase in cytoplasmic calcium levels in platelets . However, PAAmMAc particle-treated platelets demonstrated little response to thrombin in terms of an increase in {Ca2+}i and ATP release, suggesting the possibility that PAAmMAc surfaces may regulate {Ca2+}i by influencing platelet metabolism . Furthermore, sodium azide-treated platelets showed an increase in {Ca2+}i in platelets when contacting PAAmMAc particles, supporting the suggestion that PAAmMAc surfaces could regulate platelet functions . Fluorescence polarization measurements using 1,6-diphenyl-1,3,5-hexatriene-loaded platelets revealed that PAAmMAc particles increased membrane fluidity in platelets, which may be due to physicochemical interaction with PAAmMAc surfaces. New Microbiol, 1993 Jan, 16(1), 63 - 71 Non-transmissible and self-transmissible plasmids conferring drug resistance in clinical isolates of Serratia marcescens from hospitals in Riyadh, Saudi Arabia; al-Harithy RN et al.; Sixty Serratia marcescens isolates were obtained from patient specimens from three different hospitals in the city of Riyadh . These were tested for their antibiotic resistance factors using eleven different antibiotics . Their ability to transfer antibiotic resistance plasmids to a sensitive E . coli strain (RR1) was tested by transformation and conjugation experiments . Agarose gel electrophoresis was used to determine the size and number of the R-plasmids . Southern blotting was used to assess homologies between antibiotic resistance plasmids from different isolates . Among the isolates tested, 36.7% contained plasmids, and all these were from strains isolated from two hospitals . No R-plasmids could be detected among the multiple resistant strains isolated from the third hospital . Among the strains that contained plasmids, approximately 63.6% transferred multiple antibiotic resistance to E . coli and the rest transferred only one antibiotic resistance marker . The majority of strains carrying out the plasmids showed similarities in band number and size . In view of the similarities this group was denoted the predominant group, and selected for further molecular investigations . Restriction endonuclease digests of plasmids from this group gave the same restriction pattern which confirmed that they were closely related . Hybridization experiments using these plasmids and nick-translated 32p-labelled pBR-322 DNA probe, showed that all the large bands (36 kb) are related and exhibit homology with pBR-322. J Antimicrob Chemother, 1993 Jan, 31(1), 21 - 8 Alteration in expression of Serratia marcescens porins associated with decreased outer membrane permeability; Hashizume T et al.; The porin expression of two clinical isolates of Serratia marcescens, which overproduced cephalosporinase and had decreased outer membrane permeability, were studied in comparison with those of reference strains . Separation of the porin proteins assessed by SDS-PAGE containing urea revealed that both clinical isolates overexpressed a single porin of 44 and 43 kilodalton (kDa), respectively . In contrast, the in-vitro porin deficient mutant, which was derived from the reference strain IFO3736 as latamoxef-resistant, showed decreased outer membrane permeability, but produced low levels of all three peptidoglycan associated porins of 45, 44 and 43 kDa, and overexpressed 39 kDa OmpA protein . These observations suggested that the clinical isolates had a different mechanism of latamoxef resistance compared with the mutant and overexpressed possible narrow transport channels of 44 or 43 kDa . The 45 kDa porin may facilitate a more effective channel than the other two proteins . Heterogeneity of porin profiles between biotypes was also suggested . The two isolates were also resistant to penicillins and cephalosporins tested, but imipenem was the most active agent and inhibited the isolates at 1.56 mg/L. Can J Microbiol, 1993 Jan, 39(1), 108 - 11 Outer membrane proteins from Serratia marcescens; Puig M et al.; The outer membrane proteins (OMPs) of several strains of Serratia marcescens have been studied by sodium dodecyl sulphate - urea - polyacrylamide gel electrophoresis . Four major OMPs, named Omp1, Omp2, Omp3, and OmpA (42, 40, 39, and 37 kDa, respectively), have been visualized . The relative proportions of Omp2 and Omp3 depend on cultural conditions (temperature of incubation, osmolarity, and nutrient availability). Appl Environ Microbiol, 1993 Jan, 59(1), 183 - 8 Adaptation and growth of Serratia marcescens in contact lens disinfectant solutions containing chlorhexidine gluconate; Gandhi PA et al.; Serratia marcescens (11 of 12 strains) demonstrated an ability to grow in certain chlorhexidine-based disinfecting solutions recommended for rigid gas-permeable contact lenses . For a representative strain, cells that were grown in nutrient-rich medium, washed, and inoculated into disinfecting solution went into a nonrecoverable phase within 24 h . However, after 4 days, cells that had the ability to grow in the disinfectant (doubling time, g = 5.7 h) emerged . Solutions supporting growth of S . marcescens were filter sterilized . These solutions, even after removal of the cells, showed bactericidal activity against Pseudomonas aeruginosa and a biphasic survival curve when rechallenged with S . marcescens . Adaptation to chlorhexidine by S . marcescens was not observed in solutions formulated with borate ions . For chlorhexidine-adapted cells, the MIC of chlorhexidine in saline was eightfold higher than that for unadapted cells . Cells adapted to chlorhexidine showed alterations in the proteins of the outer membrane and increased adherence to polyethylene . Cells adapted to chlorhexidine persisted or grew in several other contact lens solutions with different antimicrobial agents, including benzalkonium chloride. J Bacteriol, 1993 Jan, 175(1), 176 - 81 Cloning, sequence, and expression of a chitinase gene from a marine bacterium, Altermonas sp . strain O-7; Tsujibo H et al.; The gene encoding an extracellular chitinase from marine Alteromonas sp . strain O-7 was cloned in Escherichia coli JM109 by using pUC18 . The chitinase produced was not secreted into the growth medium but accumulated in the periplasmic space . A chitinase-positive clone of E . coli produced two chitinases with different molecular weights from a single chitinase gene . These proteins showed almost the same enzymatic properties as the native chitinase of Alteromonas sp . strain O-7 . The N-terminal sequences of the two enzymes were identical . The nucleotide sequence of the 3,394-bp SphI-HindIII fragment that included the chitinase gene was determined . A single open reading frame was found to encode a protein consisting of 820 amino acids with a molecular weight of 87,341 . A putative ribosome-binding site, promoter, and signal sequence were identified . The deduced amino acid sequence of the cloned chitinase showed sequence homology with chitinases A (33.4%) and B (15.3%) from Serratia marcescens . Regardless of origin, the enzymes of the two bacteria isolated from marine and terrestrial environments had high homology, suggesting that these organisms evolved from a common ancestor. Crit Rev Microbiol, 1993, 19(2), 117 - 35 Fractal morphogenesis by a bacterial cell population; Matsuyama T et al.; Many species of bacteria have been found to form fractal colonies . Environmental (physicochemical) and biological factors for this fractal morphogenesis have been examined for their roles in the genesis of fractal and pattern diversity . Morphology of a bacterial colony on a solid agar medium depends on the nutrient diffusion field (two-dimensional) . When concentrations of nutrients are low, point-inoculated bacteria (e.g., Bacillus subtilis) exert diffusion-limited growth . A self-similar fractal colony formed slowly under such a condition has the same morphology as one made by the diffusion-limited aggregation (DLA) model . The value of fractal dimensions (ca . 1.72) and the appearance of specific phenomena (screening and repulsion effects) are consistent with computer simulations of the DLA fractal model . On the other hand, a round colony recognized on an agar-rich medium was considered to be the product of reaction-limited growth and was simulated by the Eden model . When motile bacteria are point inoculated onto semi-solid agar media, bacterial spreading behavior also is morphogenic . Branching patterns with various morphologies (e.g., dense-branching morphology) have been recognized and examined for factors responsible for pattern changes . By microscopic inspection of the extending branch, multicellular behavior of bacteria has been observed in the structured cell distribution . Besides cell division and translocation activities, wetting agents produced by some species of bacteria (e.g., serrawettins produced by Serratia marcescens) are considered to be important microbial factors for efficient space occupation and specific cell transpositions in various surface environments. J Basic Microbiol, 1993, 33(2), 123 - 30 Involvement of two genes of superinfecting phage kappa in curing and induction of prophage psi in Serratia marcescens HY; Steiger H; Prophage psi carried along with prophage y by Serratia marcescens HY is subject of moderate curing at heteroimmune superinfection of cells from stationary phase with phage kappa . Curing becomes considerably more frequent when the bacteria are non-lysogenic for y . Both psi, y-double-lysogenic and psi-single-lysogenic cells with a mutation in the ink gene are very efficiently cured of psi if infected by kappa tay, although this mutant was characterized as being deficient in transactivation of certain genes in prophage y . On the other hand to get efficiently cured after kappa wild-type infection these cells too must be devoid of a y prophage . Thus a y function turned on by tay+ seems to counteract the elimination of psi . However, interestingly enough psi curing is boosted by a further y function under special circumstances . Efficient curing depends on an intact kappa tap gene, a gene reported to cause transactivation of certain psi genes . Curing at kappa tay infection is specifically accompanied by induction of the psi prophage in a part of the infected cells . However, there is no such induction at kappa wild-type infection, either in the absence or presence of a y prophage . An explanation of these findings is suggested which includes an antirepressive effect exerted on psi and a hypothetical interaction between the products of genes tap and tay. Microbios, 1993, 76(308), 189 - 96 The role of O-antigen in susceptibility of Serratia marcescens to non-immune serum; Palomar J et al.; Selection of phage-resistant strains to three different phages, kappa, FSB3 and FSB55, whose primary receptor was lipopolysaccharide, enabled several spontaneous O-defective mutants to be isolated and characterized . Strains ATCC 274 (0:14), 2170 (0:14) and NIMA (0:6) showed serum resistance, whereas their O-side chain defective mutants exhibited high serum susceptibility . Both alternate and classical activation pathways were involved in serum killing activity in Serratia marcescens. Ren Fail, 1993, 15(5), 567 - 71 Effect of prednisolone on renal scarring in rats following infection with Serratia marcescens; Haraoka M et al.; Renal scarring is considered a criterion of reflux nephropathy and the end stage of pyelonephritis . Prednisolone, a strong anti-inflammatory drug, at doses of 1 or 2 mg/kg prevented renal scarring in rats following infection with Serratia marcescens . Four or 8 mg/kg of prednisolone, however, did not inhibit renal scar formation . In a time course experiment, renal scarring was prevented when 4-day treatment with prednisolone was initiated 2, 5, or 13 days after infection . These results show that prednisolone is effective in preventing such scarring and suggest the clinical use of this drug for preventing renal scar formation after pyelonephritis and reflux nephropathy. Postgrad Med J, 1993, 69 Suppl 3, S70 - 7 Review of disinfectant susceptibility of bacteria isolated in hospital to commonly used disinfectants; Shiraishi T et al.; The susceptibility of clinical isolates and indigenous bacteria to commonly used disinfectants was investigated during different time periods . Among the clinical isolates tested during Period I (August 1985-July 1986, 6 genera, 9 species, 353 strains) there were many resistant strains not killed within a short period of time by the recommended concentration of chlorhexidine gluconate (CHG) or benzalkonium chloride (BAC) . During Period II (October 1987-May 1988, 6 genera, 9 species, 152 strains), however, a reduction in the number of strains resistant to these disinfectants was observed . The use of the broad spectrum disinfectant povidone-iodine (PVP-I) increased between those two time periods . With regard to the susceptibility of indigenous bacteria, tests were carried out on bacteria isolated from sinks and physicians' hands in the gastroenterology division of the Departments of Internal Medicine and Surgery at the hospital . During Phase I (April-June 1987), strains of Pseudomonas and Serratia resistant to CHG and BAC were isolated from sinks, while the same strains of Serratia were also isolated from physicians' hands . During Phase II (March-May 1988), however, no resistant strains were isolated . A comparison of the consumption of disinfectants during the two phases revealed that a greater amount of CHG was consumed during Phase I, while a greater amount of PVP-I was consumed during Phase II . There was a strong indication, therefore, that bacteria resistant to CHG and BAC decrease with the increased use of PVP-I. Med Oncol Tumor Pharmacother, 1993, 10(4), 145 - 58 Clinical results and immunologic effects of a mixed bacterial vaccine in cancer patients; Havas HF et al.; A biological response modifier, mixed bacterial vaccine (MBV), derived from Streptococcus pyogenes and Serratia marcescens was used as a single agent in the treatment of 11 patients with refractory malignancies . MBV's effect on interleukin-2 (IL-2) production, plasma interferon (IFN) and tumor necrosis factor (TNF) levels was monitored . Most patients' peripheral blood mononuclear cells continued to produce baseline to elevated levels of IL-2, in spite of age and disease status . Several patients maintained moderate to high IFN levels . In general there was little correlation between IL-2 and IFN levels or with the response to therapy . One of 11 patients had minor response, 1 of 11 had partial response, 4 of 11 had temporary stabilization of disease, and 5 of 11 had progressive disease . A patient with AIDS and Kaposi's sarcoma experienced a dramatic improvement in performance status and disease stabilization . In all patients side effects occurred only following i.v . and not i.m . administration and included fever and chills . No adverse hepatic, renal or hematologic effects were observed . MBV is a well-tolerated biological response modifier with modest activity in advanced human tumors. Br J Ophthalmol, 1992 Dec, 76(12), 764 - 5 Pink hypopyon: a sign of Serratia marcescens endophthalmitis; al Hazzaa SA et al.; Endogenous bacterial endophthalmitis in infants is uncommon . We recently examined and treated an infant who presented with pink hypopyon which followed a Serratia marcescens septicaemia . Culture of the aspirate from the anterior chamber showed no red blood cells, and grew Serratia marcescens, which was also isolated from the tip of the child's umbilical artery catheter . The presence of a pink hypopyon in the absence of hyphaema may suggest the diagnosis of