Farmaco, 2002 Sep, 57(9), 715 - 22 Synthesis of halogen derivatives of benzo{h}chromene and benzo{a}anthracene with promising antimicrobial activities; Khafagy MM et al.; The synthesis of novel 7-(4-halophenyl)-8,9-dihydro-7H-12-oxa-9,11-diaza-benzo{a}anthracene derivatives has been reported . The key intermediate 3-amino-9-chloro-1-(4-halophenyl)-1H-benzo{h}chromene-2-carbonitrile (3) was obtained by treating 4-halobenzylidenmalononitriles (1a-c) and ethyl 4-halobenzylidenmalonates (1d-f) with 4-chloro-1-naphthol (2) in ethanolic piperidine solution . Antimicrobial activity was shown for most of the synthesized compounds. Antimicrob Agents Chemother, 2002 Nov, 46(11), 3585 - 90 Cationic hydrophobic peptides with antimicrobial activity; Stark M et al.; The MICs of cationic, hydrophobic peptides of the prototypic sequence KKAAAXAAAAAXAAWAAXAAAKKKK-amide (where X is one of the 20 commonly occurring amino acids) are in a low micromolar range for a panel of gram-negative and gram-positive bacteria, with no or low hemolytic activity against human and rabbit erythrocytes . The peptides are active only when the average segmental hydrophobicity of the 19-residue core is above an experimentally determined threshold value (where X is Phe, Trp, Leu, Ile, Met, Val, Cys, or Ala) . Antimicrobial activity could be increased by using peptides that were truncated from the prototype length to 11 core residues, with X being Phe and with 6 Lys residues grouped at the N terminus . We propose a mechanism for the interaction between these peptides and bacterial membranes similar to the "carpet model," wherein the Lys residues interact with the anionic phospholipid head groups in the bacterial membrane surface and the hydrophobic core portion of the peptide is then able to interact with the lipid bilayer, causing disruption of the bacterial membrane. J Surg Res, 2002 Sep, 107(1), 7 - 13 Systemic inflammatory response induced by dacron graft and modulation by antimicrobial agents: experimental study; Lozano FS et al.; BACKGROUND: The purpose of this work was to evaluate the effect that different antimicrobial agents and different forms of administering them would have over a systemic inflammatory response (SIR) induced by an intraperitoneally implanted collagen-coated Dacron graft . MATERIALS AND METHODS: Thirty-six male Wistar rats were randomly allocated into six groups of 6 animals each: (I) control, (II) "sham," (III) graft but no antibiotic, (IV) graft plus systemic cefazolin, (V) graft plus locally applied gentamicin, and (VI) graft soaked in rifampicin.After 72 h, mesenteric lymph nodes, liver, kidney, and the implanted graft were sent to the microbiology laboratory and cultured for aerobic and anaerobic organisms in order to evaluate bacterial translocation . Serum cytokines (IL-1beta and TNF-alpha), myeloperoxidase activity in liver and kidney, and superoxide anion and superoxide dismutase activities in liver were also determined to evaluate the level of SIR . RESULTS: Microbiologic and biochemical data indicated that intraperitoneal implantation of a collagen-coated Dacron graft induced a significant (P < 0.05) bacterial translocation and a high inflammatory response, both of which decreased significantly with antibiotic treatment regardless of the means of administration (P < 0.05) . CONCLUSIONS: The present experimental model shows that the antibiotics used, in different means of administration, reduce bacterial translocation and behave as modulators of the SIR induced by an intraperitoneal collagen-coated Dacron graft. Lancet Infect Dis, 2002 Oct, 2(10), 593 - 604 Bacterial eradication in the treatment of otitis media; Dagan R et al.; Drugs differ in their ability to eradicate various pathogens from the middle-ear cavity during acute otitis media (AOM), and these differences clearly affect clinical outcome . Outcome is derived from differences in the association between concentrations of the drugs at the site of infection and the antimicrobial effect (termed pharmacodynamics) . These differences are even more marked in the present era of antimicrobial resistance . However, since AOM is a self-limiting disease in most cases, difference in clinical outcome is more difficult to ascertain than that of bacteriological outcome, which is measured within 3-5 days . A favourable clinical outcome regardless of the bacteriological effect of the drug can result in false optimism when less-effective antibiotic drugs are used . Inappropriate study design and manipulation of clinical results add to this confusion . In this review we attempt to highlight the evidence regarding bacteriological response to antibiotics in AOM and to draw attention to potential flaws that may mislead clinicians. J Clin Pharm Ther, 2002 Oct, 27(5), 353 - 5 Quantification of Helicobacter pylori resistance in functional and organic dyspepsia; Rosandic M et al.; OBJECTIVE: To compare the efficacy of Helicobacter pylori eradication in patients with functional and organic dyspepsia . METHODS: The study included a cohort of 160 patients (115 with organic and 45 with functional dyspepsia) with dyspeptic symptoms and gastroscopically confirmed H . pylori infection . Triple therapy with omeprazole 20 mg, amoxicillin 1000 mg and metronidazole 400 mg (OAM) was administered twice a day for a week . Minimal inhibition concentration (MIC) was estimated on cultures from 41 patients with positive H . pylori for determination of antimicrobial sensitivity and primary resistance to amoxicillin and metronidazole . RESULTS: Endoscopic examination at least 6 weeks after therapy showed that 116 (72.5%) patients had H . pylori eradicated, whereas 44 (27.5%) were not . From the latter patients, 10 (23%) had functional dyspepsia and from 116 eradicated patients 35 (30%) had functional dyspepsia . Difference in efficacy of OAM therapy between patients with organic and functional dyspepsia was not significant (P > 0.5) . Percentages of non-eradicated patients with organic and functional dyspepsia were 29.6 and 22.2%, respectively (ratio 1.3 : 1) . MIC from 41 samples showed 18 (44%) in vitro resistant strains . There was no resistance to amoxicillin . CONCLUSIONS: There is no significant difference in H . pylori resistance to the same antibiotic between patients having functional or organic dyspepsia. Drugs Aging, 2002, 19(9), 647 - 69 HIV disease and advanced age: an increasing therapeutic challenge; Manfredi R; The mean age of patients at both first HIV detection and AIDS diagnosis is progressively rising over time . However, reliable epidemiological estimates, clinical data or controlled therapeutic and outcome figures are lacking for elderly patients, especially with regard to laboratory and clinical response to antiretroviral therapy, treatment tolerability, drug-drug interactions, short- and long-term toxicity, and interactions with underlying illnesses and concurrent pharmacological treatment . In fact, the large majority of randomised, controlled trials evaluating and comparing new antiretroviral drugs or anti-HIV therapeutic strategies, as well as antimicrobial treatment or chemoprophylaxis of HIV-related complications, either excluded patients with advanced age and/or concurrent disorders or did not offer substudies or detailed data analysis focusing on older patients compared with younger ones . The life expectancy of HIV-infected persons receiving highly active antiretroviral therapy (HAART) is now extended (approaching that of the general population), so that the definition of AIDS has lost its epidemiological and clinical significance thanks to the immune reconstitution resulting from potent antiretroviral therapy . However, an ever-increasing number of individuals aged > or =50 years with HIV infection is expected in the coming years, as a result of both increased survival of patients with treated disease and delayed recognition of individuals with occult HIV disease . The limited data available about combined antiretroviral therapy in the elderly seem to show an overlapping virological success rate but a slower and blunted immune recovery compared with younger patients . Thymic output, however, seems somewhat preserved even in adulthood and may contribute to the reconstitution of most of the quantitative and functional T cell abnormalities caused by HIV disease . More attention must be paid to underlying end-organ disorders, as well as expected pharmacological interactions and combined drug toxicity that may interfere with HAART efficacy and patients' compliance with recommended regimens and could lead to increased adverse effects . The available guidelines for antiretroviral treatment and therapy and prophylaxis of AIDS-related illnesses should be regularly updated and should include problems related to HIV disease in an aging population . Specific trials or substudies focusing on older people are warranted to obtain controlled data on all issues of antiretroviral therapy in the elderly, including time and mode of initiation, and modification and salvage HAART regimens . Antiretroviral drug dosage adjustment to take into account underlying pathological conditions or other pharmacological treatments is another emerging issue. J Am Chem Soc, 2002 Oct 23, 124(42), 12543 - 9 Solid-phase synthesis of fullerene-peptides; Pantarotto D et al.; The solid-phase synthesis of peptides (SPPS) containing {60}fullerene-functionalized amino acids is reported . A new amino acid, fulleropyrrolidino-glutamic acid (Fgu), is used for the SPPS of a series of analogues of different length based on the natural Leu(5)-Enkephalin and on cationic antimicrobial peptides . These fullero-peptides were prepared on different solid supports to analyze the influence of the resin on the synthesis . Optimized protocols for the coupling and deprotection procedures were determined allowing the synthesis of highly pure peptides in sufficient quantities for evaluation of biological activities . In particular, to avoid side reactions of the fullerene moiety with bases and nucleophiles, the removal of the protecting groups was performed under inert conditions (nitrogen or argon in the dark) . We have encountered serious problems with the recovery of the crude compounds, especially when Fgu was inserted in the proximity of the resin core as fullero-peptides tend to remain embedded inside the resin . Eventually, all of the fullero-peptides were easily purified, and the cationic peptides were tested for their antimicrobial activities . They displayed a specific activity against the Gram-positive bacterium S . aureus and also lysed erythrocytes . The availability of a fullero-amino acid easily useable in the SPPS of fullero-peptides may thus open the way to the synthesis of new types of biologically active oligomers. J Food Prot, 2002 Oct, 65(10), 1632 - 6 Combined effects of mustard flour, acetic acid, and salt against Esherichia coil O157:H7 stored at 5 and 22 degrees C; Rhee MS et al.; The combined effects of acetic acid and mustard flour were investigated to ascertain their impact on Escherichia coli O157:H7 stored at 5 and 22 degrees C . Samples were prepared with various concentrations of acetic acid (0, 0.25, 0.5, 0.75, and 1% {vol/vol}) combined with 10% (wt/vol) Baltimore or Coleman mustard flour and 2% (fixed; wt/vol) sodium chloride . An acid-adapted mixture of three E . coli O157:H7 strains (10(6) to 10(7) CFU/ml) was inoculated into prepared mustard samples that were stored at 5 and 22 degrees C, and samples were assayed periodically for the survival of E . coli O157:H7 . The numbers of E . coli O157:H7 were reduced much more rapidly at 22 degrees C than at 5 degrees C . E . coli O157:H7 was rapidly reduced to below the detection limit (<0.3 log10, CFU/ml) after 1 day at 22 degrees C, whereas it survived for up to 5 days at 5 degrees C . There was no synergistic or additive effect with regard to the killing of E . coli O157:H7 with the addition of small amounts of acetic acid to the mustard flour . When stored at 5 degrees C, mustard in combination with 0.25 (M-0.25), 0.5 (M-0.5), and 0.75% (M-0.75) acetic acid exerted less antimicrobial activity than the control (M-0) . The order of lethality at 5 degrees C was generally M-0.25 = M-0.5 < M-0.75 = M-0 < M-1 . The addition of small amounts of acetic acid (<0.75%) to mustard retards the reduction of E coli O157:H7 . Statistical reduction in populations of E . coli O157:H7 (P < 0.05) was enhanced relative to that of the control (mustard alone) only with the addition of 1% acetic acid . This information may help mustard manufacturers to understand the antimicrobial activity associated with use of mustard flour in combination with acetic acid. Clin Ther, 2002 Sep, 24(9), 1414 - 25 Open-label, randomized comparison of the efficacy and tolerability of clarithromycin, levofloxacin, and cefuroxime axetil in the treatment of adults with acute bacterial exacerbations of chronic bronchitis; Weiss LR; BACKGROUND: In the absence of a confirmed pathogen, empiric antimicrobial treatment of patients with acute exacerbations of chronic bronchitis and acute bacterial exacerbations of chronic bronchitis (ABECB) is accepted as standard practice and recommended in treatment guidelines . OBJECTIVE: This study compared the efficacy and tolerability of a 10-day course of 3 antimicrobial regimens commonly used to treat adults with ABECB . METHODS: This prospective, open-label, randomized study assessed clarithromycin 500 mg twice daily, levofloxacin 500 mg once daily, and cefuroxime axetil 250 mg twice daily, each administered for 10 days with food, in patients with ABECB . Efficacy was determined on the basis of the clinical response to treatment and need for hospitalization and/or further antimicrobial therapy . RESULTS: A total of 283 patients (150 men, 133 women) with a mean age of 55 years (range, 29 to 86 years) were randomized to receive clarithromycin (n = 97), levofloxacin (n = 94), or cefuroxime axetil (n = 92) . Of 262 clinically assessable patients, clinical cure or improvement occurred in 87.9% (80/91) of those treated with clarithromycin, 87.4% (76/87) of those treated with levofloxacin, and 79.8% (67/84) of those treated with cefuroxime axetil . Eight (8.8%) clarithromycin-treated patients, 6 (6.9%) levofloxacin-treated patients, and 12 (14.3%) cefuroxime axetil-treated patients required a change in antimicrobial therapy to achieve clinical cure/improvement; between-group differences were not significant . No patients treated with clarithromycin required hospitalization for further antimicrobial treatment, compared with 3.4% (3/87) of levofloxacin-treated and 3.6% (3/84) of cefuroxime axetil-treated patients (P = NS) . A total of 6.2% (6/97) of clarithromycin-treated patients were prematurely discontinued from treatment due to adverse events, compared with 7.4% (7/94) and 8.7% (8/92) of levofloxacin- and cefuroxime axetil-treated patients, respectively . CONCLUSION: A high rate of clinical efficacy and tolerability was observed in this population of patients with ABECB treated with clarithromycin 500 mg twice daily, levofloxacin 500 mg once daily, or cefuroxime axetil 250 mg twice daily for 10 days. Vet Clin North Am Small Anim Pract, 2002 Sep, 32(5), 1101 - 26 Nosocomial infections; Johnson JA; Nosocomial infections and antimicrobial resistance are topics that have been intensely studied in human medicine because of their significant impact on human health . In recent years, concerns have been raised that the use of antibiotics in veterinary medicine, animal husbandry, and agriculture may be contributing to the development of resistance in common bacterial species affecting human beings . Although there is inadequate proof at this time that the resistance is transmitted from animals to people, if antibiotics continue to be used indiscriminately in veterinary medicine, veterinarians may find themselves facing regulations restricting the use of some antibiotics . Nosocomial infections have been reported in veterinary medicine and are likely to increase in prevalence with the increase in intensive care practices in many hospitals . Prolonged hospitalization and the use of invasive devices and procedures increase the risk of nosocomial disease . As in human medicine, organisms isolated in the nosocomial infections reported in veterinary patients have an increasingly broad spectrum of antimicrobial resistance . Despite these findings, the use of empiric and prophylactic antibiotic therapy is still widespread in veterinary medicine . Nosocomial infections and antimicrobial resistance may have a serious impact on the future of {table: see text} veterinary medicine, because the cost and ability to treat our patients may be affected by the loss of access to or effectiveness of antimicrobial drugs . Despite the millions of dollars spent on research to reduce the incidence of nosocomial infections in human patients, the strategies that have consistently proven successful are simple and inexpensive to implement . The most important factor in preventing nosocomial infections is improving the hygiene practices of health care providers . Hand-washing or the use of disposable gloves can dramatically reduce the transmission of bacteria between patients . Aseptic technique should be used in the placement and management of all invasive devices . All staff members should be educated on the risks and symptoms associated with nosocomial infections so that cases can be detected early and treated appropriately . We in the veterinary profession have the opportunity to learn from the experiences of the human medical profession and can take steps to prevent the escalation of nosocomial infections and their impact on our profession. J Pharm Sci, 2002 Nov, 91(11), 2452 - 64 Activity-bioavailability balance in oral drug development for a selected group of 6-fluoroquinolones; Talens-Visconti R et al.; A nomogram is proposed to select the best candidate in drug development studies with quinolones and is intended to substitute other possible models . The nomogram is referred to as an activity-bioavailability balance (ABB) because it includes the following two criteria: ABB = {(1/gm MIC drug candidate)/ (1/gm MIC ciprofloxacin)}.{(F(calc) drug candidate)/( F(calc) ciproflaxacin)} . The in vitro activity of a group of 4'N-alkyl-ciprofloxacin derivatives was determined together with that of ciprofloxacin, initially against some reference strains and subsequently against 159 clinical isolates of eight selected species . The inverse of the geometric mean of the lowest concentration of drug at which the original inoculum was reduced to no more than two colonies (1/gm MIC), as an antimicrobial activity parameter, and the absolute oral bioavailability index (F(calc)), as predicted from in situ intestinal absorption rate constants, were used for calculation of the ABB values, which ranged from 0.1 to 17 for the species and compounds tested . Ciprofloxacin was the best candidate only against Escherichia coli, whereas 4'N-methyl- and/or 4'N-ethyl-ciprofloxacin showed better or much better ABB values than the model drug, and can be selected as potential drug candidates against the remaining clinical strains . The procedure described could be a useful technique for further drug development studies . J Pharm Sci, 2002 Nov, 91(11), 2283 - 300 Excipient-drug interactions in parenteral formulations; Akers MJ; Excipients are added to parenteral formulations to enhance or maintain active ingredient solubility (solubilizers) and/or stability (buffers, antioxidants, chelating agents, cryo- and lyoprotectants) . Excipients also are important in parenteral formulations to assure safety (antimicrobial preservatives), minimize pain and irritation upon injection (tonicity agents), and control or prolong drug delivery (polymers) . These are all examples of positive or synergistic interactions between excipients and drugs . However, excipients may also produce negative effects such as loss of drug solubility, activity, and/or stability . This review article will highlight documented interactions, both synergistic and antagonistic, between excipients and drugs in parenteral formulations . The reader will gain better understanding and appreciation of the implications of adding formulation ingredients to parenteral drug products . Infect Immun, 2002 Nov, 70(11), 6302 - 9 Activation of NKT cells protects mice from tuberculosis; Chackerian A et al.; The T-cell immune response to Mycobacterium tuberculosis is critical in preventing clinical disease . While it is generally accepted that both major histocompatibility complex class I (MHC-I)-restricted CD8(+) and MHC-II-restricted CD4(+) T cells are important for the immune response to M . tuberculosis, the role of non-MHC-restricted T cells is still not clearly delineated . We have previously reported that CD1d(-/-) mice do not differ from CD1d(+/+) mice in their survival following infection with M . tuberculosis . We now show that, although CD1d-restricted NKT cells are not required for optimum immunity to M . tuberculosis, specific activation of NKT cells by the CD1d ligand alpha-galactosylceramide protects susceptible mice from tuberculosis . Treatment with alpha-galactosylceramide reduced the bacterial burden in the lungs, diminished tissue injury, and prolonged survival of mice following inoculation with virulent M . tuberculosis . The capacity of activated NKT cells to stimulate innate immunity and modulate the adaptive immune response to promote a potent antimicrobial immune response suggests that alpha-galactosylceramide administration could have a role in new strategies for the therapy of infectious diseases. J Biol Chem, 2002 Dec 20, 277(51), 49332 - 40 Epub 2002 Oct 11. Dermaseptins from Phyllomedusa oreades and Phyllomedusa distincta . Anti-Trypanosoma cruzi activity without cytotoxicity to mammalian cells; Brand GD et al.; Amphibian skin secretions are known as a rich source of biologically active molecules, most of which are alkaloids, biogenic amines, and peptides . Dermaseptins are a class of antimicrobial peptides present in tree frogs of the Phyllomedusa genus . They are cationic molecules of 28-34 residues that permeabilize the membrane of Gram-positive and Gram-negative bacteria, yeasts, and filamentous fungi, showing little or no hemolytic activity . This work reports the isolation, molecular mass analysis, primary structure determination, biological activities, and potential therapeutic applications of an antimicrobial peptide found in the skin secretion of Phyllomedusa oreades, which is a newly described amphibian species endemic of the Brazilian savanna . DS 01 is a 29-residue-long peptide with a molecular mass of 2793.39 Da showing antibacterial properties against Gram-positive and Gram-negative bacteria in the range of 3-25 microm . Anti-protozoan activity was investigated using T . cruzi in its trypomatigote and epimastigote forms cultivated in both cell culture and blood media . Within 2 h after incubation with DS 01 at a final concentration of approximately 6 microm, no protozoan cells were detected . Two synthetic dermaseptins, described previously by our group and named dermadistinctins K and L (DD K and DD L), also had their anti-Trypanosoma cruzi activity investigated and demonstrated similar properties . Toxicity of DS 01 to mouse erythrocytes and white blood cells was evaluated by means of atomic force microscopy and flow cytometry . No morphological alterations were observed at a lytic concentration of DS 01, suggesting its therapeutic value especially as an anti-T . cruzi agent to prevent infections during blood transfusion. J Immunol Methods, 2002 Dec 1, 270(1), 53 - 62 Detection of dermcidin-derived peptides in sweat by ProteinChip technology; Flad T et al.; Recently, a novel antimicrobial peptide DCD-1, derived from the Dermcidin (DCD) gene and secreted by sweat glands, has been described by Schittek et al . {Nat . Immunol . 2 (2001) 1133.} . Here we describe the application of the surface-enhanced laser desorption/ionisation (SELDI) technology for the detection of DCD-1 and other dermcidin-derived peptides directly from microlitre amounts of human sweat . The advantages of the technique are as follows: (a) it can be carried out with ease and rapidity; (b) multiple samples can be processed simultaneously; (c) prior purification is not required; and (d) only a limited sample volume is necessary for both protein profiling and semiquantitation . Profiling of human sweat from various donors revealed that in addition to DCD-1, other DCD-derived peptide species were also present in significant quantities . Four of five identified peptides were DCD-1 related, while the fifth corresponded to a portion of the DCD protein outside the DCD-1 core . This provides clues as to how the novel protein is processed to its active form, though further work remains to elucidate this fully . Thus, we have demonstrated the applicability of such technology to the detection of DCD-1 and for the protein profiling of sweat in general . Such studies could reveal valuable new biomarkers for diagnosis and treatment of skin and sweat gland disorders. Biochemistry, 2002 Oct 22, 41(42), 12835 - 42 Design of non-cysteine-containing antimicrobial beta-hairpins: structure-activity relationship studies with linear protegrin-1 analogues; Lai JR et al.; Protegrins are short, cationic peptides that display potent, broad-spectrum antimicrobial activity . PG-1, the first of the five natural analogues discovered, forms a rigid antiparallel two-stranded beta-sheet that is stabilized by two disulfide bonds . The two strands of the sheet are linked by a short two-residue loop segment . Removal of the disulfide bridges (e.g., in Cys --> Ala analogues) is known to cause marked loss of antimicrobial activity . We have used basic principles of beta-hairpin design to develop linear analogues of PG-1 that lack cysteine but nevertheless display PG-1-like activity . Our most potent reengineered molecules contain three essential design features: (i) the four cysteine residues of PG-1 are replaced by residues that have high propensity for beta-strand conformation, (ii) D-proline is placed at the i + 1 position of the reverse turn to promote a type II' beta-turn, and (iii) amino functionality is incorporated at the gamma-carbon of the D-proline residue to mimic the charge distribution of the natural beta-hairpin . Structural studies revealed that the antimicrobial potency of the non-disulfide-bonded peptides can be correlated to the stability of the beta-hairpin conformations they adopt in aqueous solution . The presence of 150 mM NaCl was found to have little effect on the antimicrobial activity of PG-1, but one of our linear analogues loses some potency under these high salt conditions . Despite this discrepancy in salt sensitivity, NMR and CD data indicate that neither PG-1 nor our linear analogue experiences a significant decrease in beta-hairpin conformational stability in the presence of 150 mM NaCl . Thus, salt inactivation is not due to destabilization of the beta-hairpin conformation . Furthermore, our results show that beta-sheet design principles can be used to replace conformation-stabilizing disulfide bridges with noncovalent conformation-stabilizing features. Biotechnol Bioeng, 2002 Dec 20, 80(6), 599 - 609 Electro-membrane filtration for the selective isolation of bioactive peptides from an alpha(s2)-casein hydrolysate; Bargeman G et al.; For the isolation of the ingredients required for functional foods and nutraceuticals generally membrane filtration has too low a selectivity and chromatography is (too) expensive . Electro-membrane filtration (EMF) seems to be a breakthrough technology for the isolation of charged nutraceutical ingredients from natural sources . EMF combines the separation mechanisms of membrane filtration and electrophoresis . In this study, positively charged peptides with antimicrobial activity were isolated from an alpha(s2)-casein hydrolysate using batch-wise EMF . alpha(s2)-Casein f(183-207), a peptide with strong antimicrobial activity, predominated in the isolated product and was enriched from 7.5% of the total protein components in the feed to 25% in the permeate product . With conventional membrane diafiltration using the same membrane (GR60PP), isolation of this and other charged bioactive peptides could not be achieved . The economics of EMF are mainly governed by the energy costs and the capital investment, which is affected by the flux of the desired peptide . A maximum average transport rate of alpha(s2)-casein f(183-207) during batch-wise EMF of 1.2 g/m2 . h was achieved . Results indicate that an increase in the hydrolysate (feed) concentration, the applied potential difference and the conductivity of the permeate and electrode solutions, and a reduction in the conductivity of the feed result in a higher transport rate of alpha(s2)-casein f(183-207) . This is in line with the expectation that the transport rate is improved when the concentration, the electrical field strength, or the electrophoretic mobility is increased, provided that the electrophoretic transport predominates . The expected energy consumption of the EMF process per gram of peptide transported was reduced by approximately 50% by applying a low overall potential difference and by processing desalinated hydrolysate . Considerable improvements in transport rate, energy efficiency, and process economics seem to be attainable by additional optimization of the process parameters and the EMF module design . J Leukoc Biol, 2002 Oct, 72(4), 677 - 84 Calcium spikes in activated macrophages during Fcgamma receptor-mediated phagocytosis; Myers JT et al.; Rises in intracellular-free calcium ({Ca(2+)}(i)) have been variously associated with Fcgamma receptor (FcR)-mediated phagocytosis in macrophages . We show here that activation of murine bone marrow-derived macrophages increases calcium spiking after FcR ligation . Ratiometric fluorescence microscopy was used to measure {Ca(2+)}(i) during phagocytosis of immunoglobulin G (IgG)-opsonized erythrocytes . Whereas 13% of nonactivated macrophages increased {Ca(2+)}(i) in the form of one or more spikes, 56% of those activated with lipopolysaccharides (LPS; 18 h at 100 ng/ml) and interferon-gamma (IFN-gamma; 100 U/ml) and 73% of macrophages activated with LPS, IFN-gamma, interleukin (IL)-6 (5 ng/ml), and anti-IL-10 IgG (5 micro g/ml) spiked calcium during phagocytosis . Calcium spikes were inhibited by thapsigargin (Tg), indicating that they originated from endoplasmic reticulum . The fact that activated macrophages showed a more dramatic response suggested that calcium spikes during phagocytosis mediate or regulate biochemical mechanisms for microbicidal activities . However, lowering {Ca(2+)}(i) with ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid or inhibiting calcium spikes with Tg did not inhibit phagosome-lysosome fusion or the generation of reactive oxygen or nitrogen species . Thus, the increased calcium spiking in activated macrophages was not directly associated with the mechanism of phagocytosis or the increased antimicrobial activities of activated macrophages. Trends Microbiol, 2002, 10(10 Suppl), S8 - 14 Microbial and viral drug resistance mechanisms; McKeegan KS et al.; Microorganisms and viruses have developed numerous resistance mechanisms that enable them to evade the effect of antimicrobials and antivirals . As a result, many have become resistant to almost every available means of treatment . This problem, although not new, is becoming increasingly acute and it is now clear that a fundamental understanding of the mechanisms that microbes and viruses deploy in the development of resistance is essential if we are to gain new insights into ways to combat this problem. Structure (Camb), 2002 Oct, 10(10), 1363 - 70 Structure of the cathelicidin motif of protegrin-3 precursor: structural insights into the activation mechanism of an antimicrobial protein; Sanchez JF et al.; Cathelicidins are a family of antimicrobial proteins isolated from leucocytes and epithelia cells that contribute to the innate host defense mechanisms in mammalians . Located in the C-terminal part of the holoprotein, the cathelicidin-derived antimicrobial peptide is liberated by a specific protease cleavage . Here, we report the X-ray structure of the cathelicidin motif of protegrin-3 solved by MAD phasing using the selenocysteine-labeled protein . Its overall structure represents a fold homologous to the cystatin family and adopts two native states, a monomer, and a domain-swapped dimer . This crystal structure is the first example of a structural characterization of the highly conserved cathelicidin motif and thus provides insights into the possible mechanism of activation of the antimicrobial protegrin peptide. Diagn Microbiol Infect Dis, 2002 Sep, 44(1), 69 - 76 Fluoroquinolones for the treatment of outpatient community-acquired pneumonia; Jones RN et al.; The increasing prevalence of beta-lactam and macrolide resistance in bacteria that cause respiratory infections has underscored the need for effective antimicrobial agents . The broad spectrum, excellent oral bioavailability, and once-daily dosing of fluoroquinolones contributed to the introduction of several new agents in the past decade . This class is among the world's most used antimicrobial therapies in community and hospital settings . Fluoroquinolones are generally well tolerated, but safety profiles differ widely among agents . Knowledge of in vitro activity, local microbiologic susceptibility and resistance patterns, adverse effects, and potential drug interactions should influence the selection of the best agent for individual patients . This overview of the fluoroquinolones directs particular attention to use in community-acquired pneumonia and safety. J Pathol, 2002 Nov, 198(3), 369 - 77 Antimicrobial peptides are expressed and produced in healthy and inflamed human synovial membranes; Paulsen F et al.; The objective of this study was to determine the expression and production of antimicrobial peptides by healthy and inflamed human synovial membranes . Deposition of the antimicrobial peptides lysozyme, lactoferrin, secretory phospholipase A(2) (sPA(2)), matrilysin (MMP7), human neutrophil alpha-defensins 1-3 (HNP 1-3), human beta-defensin 1 (HBD-1), and human beta-defensin 2 (HBD-2) was determined by immunohistochemistry . Expression of mRNA for the antimicrobial peptides bactericidal permeability-increasing protein (BPI), heparin binding protein (CAP37), human cationic antimicrobial protein (LL37), human alpha-defensin 5 (HD5), human alpha-defensin 6 (HD6), HBD-1, HBD-2, and human beta-defensin 3 (HBD-3) was analysed by reverse transcription polymerase chain reaction (RT-PCR) . RT-PCR revealed CAP37 and HBD-1 mRNA in samples of healthy synovial membrane . Additionally, HBD-3 and/or LL37 mRNA was detected in synovial membrane samples from patients with pyogenic arthritis (PA), osteoarthritis (OA) or rheumatoid arthritis (RA) . BPI, HD5, HD6, and HBD-2 mRNAs were absent from all samples investigated . Immunohistochemistry identified lysozyme, lactoferrin, sPA(2), and MMP7 in type A synoviocytes of all samples . HBD-1 was only present in type B synoviocytes of some of the samples . Immunoreactive HBD-2 peptide was only visible in some inflamed samples . HNP1-3 was detected in both healthy and inflamed synovial membranes . The data suggest that human synovial membranes produce a broad spectrum of antimicrobial peptides . Under inflammatory conditions, the expression pattern changes, with induction of HBD-3 in PA (LL37 in RA; HBD-3 and LL37 in OA) as well as down-regulation of HBD-1 . HBD-3 holds therapeutic potential in PA as it has a broad spectrum of antimicrobial activity and accelerates epithelial healing . However, caution is appropriate since defensins also promote fibrin formation and cell proliferation - key elements in joint infection . Clarification of the role of antimicrobial peptides in OA and RA will require further investigation . J Antibiot (Tokyo), 2002 Aug, 55(8), 696 - 701 New dithiolopyrrolone antibiotics from Saccharothrix sp . SA 233 . I . Taxonomy, fermentation, isolation and biological activities; Lamari L et al.; Three new natural antibacterial and antifungal dithiolopyrrolone antibiotics were isolated along with the known iso-butyropyrrothine and thiolutine from the fermentation broth of an actinomycete strain which was isolated from a saharian palm grove soil collected at Adrar, south Algeria . The strain was identified as Saccharothrix sp . The three new antibiotics exhibited broad antimicrobial activity against gram-positive bacteria, yeasts and fungi in vitro. Mass Spectrom Rev, 2002 Mar-Apr, 21(2), 87 - 107 Collision-induced fragmentations of the (M-H)- parent anions of underivatized peptides: an aid to structure determination and some unusual negative ion cleavages; Bowie JH et al.; This article describes the fundamental cleavage reactions of (M-H)(-) anions of underivatized peptides that contain up to 25 amino acid residues . The experimental observations of these cleavages have been backed up by molecular modeling, generally at the AM1 level of theory . The basic cleavages are the ubiquitous alpha- and beta-backbone cleavage reactions, which provide information similar to that of the B and Y + 2 cleavages of MH(+) ions of peptides . The residues Asp and Asn also effect cleavages of the backbone (called delta- and gamma-cleavages), by reactions initiated from side chain enolate anions, causing elimination reactions that cleave the backbone between the Asp (Asn) N bond;C backbone bond . Glu and Gln also direct analogous delta- and gamma-cleavages of the backbone, but in this case the processes are initiated by attack of the side chain CO(2) (-) (CONH(-)) to form a lactone (lactam) . Ser and Thr residues undergo characteristic fragmentations of the side chain . These processes, losses of CH(2)O (Ser) and MeCHO (Thr), convert these residues into Gly . In larger peptides, Ser and Thr can effect two backbone cleavage reactions, called gamma- and epsilon -processes . The C-terminal CO(2) (-) (or CONH(-)) forms a hydrogen bond with the side chain OH (of Ser or Thr), placing the C-terminal residue in a position where it may affect S(N) (2) attack at the electrophilic backbone CH of Ser, with concomitant cleavage of the backbone . All of the above negative ion cleavages require the peptide backbone to be conformationally flexible . However, there is a backbone cleavage that requires the peptide to have an alpha-helical conformation in order for the two reacting centers to approach . This cleavage is illustrated for the Glu 23-initiated backbone cleavage at Ile 21 for the (M-H)(-) anion of the antimicrobial peptide caerin 1.1 . J Biol Chem, 2002 Dec 20, 277(51), 49921 - 6 Epub 2002 Oct 07. Epithelial innate immunity . A novel antimicrobial peptide with antiparasitic activity in the blood-sucking insect Stomoxys calcitrans; Boulanger N et al.; The gut epithelium is an essential interface in insects that transmit parasites . We investigated the role that local innate immunity might have on vector competence, taking Stomoxys calcitrans as a model . S . calcitrans is sympatric with tsetse flies, feeds on many of the same vertebrate hosts, and is thus regularly exposed to the trypanosomes that cause African sleeping sickness and nagana . Despite this, S . calcitrans is not a cyclical vector of these trypanosomes . Trypanosomes develop exclusively in the lumen of digestive organs, and so epithelial immune mechanisms, and in particular antimicrobial peptides (AMPs), may be the prime determinants of the fate of an infection . To investigate why S . calcitrans is not a cyclical vector of trypanosomes, we have looked in its midgut for AMPs with trypanolytic activity . We have identified a new AMP of 42 amino acids, which we named stomoxyn, constitutively expressed and secreted exclusively in the anterior midgut of S . calcitrans . It displays an amphipathic helical structure and exhibits a broad activity spectrum affecting the growth of microorganisms . Interestingly, this AMP exhibits trypanolytic activity to Trypanosoma brucei rhodesiense . We argue that stomoxyn may help to explain why S . calcitrans is not a vector of trypanosomes causing African sleeping sickness and nagana. Neuropeptides, 2002 Aug, 36(4), 246 - 51 Langerhans cell expression of neuropeptide Y and peptide YY; Lambert RW et al.; Neuropeptide Y (NPY) and peptide YY (PYY) are structurally related peptides with a variety of known functions . The role of these peptides in the skin is largely unknown, although NPY-like immunoreactivity has been reported in the epidermis . The recent report that these peptides have antimicrobial properties suggests that NPY and PYY may contribute to the skin's defense mechanisms against invading microorganisms . We have demonstrated that Langerhans cells (LC) and a certain BALB/c epidermis-derived dendritic cell line contain mRNA for NPY and PYY using RT-PCR . Furthermore, this dendritic cell line as well as an epidermis-derived dendritic cell line from A/J mice were found to produce NPY and PYY and LC produced PYY, as assessed by radioimmunoassay . These data suggest that the protective function of LC include not only antigen presentation, but also production of antimicrobial peptides . Bioorg Med Chem Lett, 2002 Nov 4, 12(21), 3171 - 4 The antimicrobial natural product chuangxinmycin and some synthetic analogues are potent and selective inhibitors of bacterial tryptophanyl tRNA synthetase; Brown MJ et al.; The antimicrobial natural product chuangxinmycin has been found to be a potent and selective inhibitor of bacterial tryptophanyl tRNA synthetase (WRS) . A number of analogues have been synthesised . The interaction with WRS appears to be highly constrained, as only sterically smaller analogues afforded significant inhibition . The only analogue to show inhibition comparable to chuangxinmycin also had antibacterial activity . WRS inhibition may contribute to the antibacterial action of chuangxinmycin. Enferm Infecc Microbiol Clin, 2002 Oct, 20(8), 407 - 11; quiz 412 {Pharmacokinetic and pharmacodynamic concepts for an interpretative reading of the antibiogram}; Soriano F; Both microbiological and epidemiological reading of the antibiogram can be performed without additional pharmacokinetic or pharmacodynamic concepts . However, when the aim is a clinical reading of the antibiogram, knowledge of the pharmacokinetics of antimicrobial agents and of all phenomena occurring between antimicrobial agents and microorganisms is imperative . Pharmacokinetics includes the study of absorption, distribution, metabolism and elimination of drugs . These data provide information on the active concentrations of antimicrobial agents in blood and other fluids as well as in the tissues where infection may develop . Knowledge of metabolism and of elimination pathways complete the main pharmacokinetic parameters of antimicrobial agents . The bactericidal effect of antimicrobial agents can be either concentration- or time-dependent . In the former, high concentrations of antimicrobial agents, much higher than their corresponding MICs, are needed, while in the latter, maintaining the concentration of antimicrobial agents at levels slightly higher than their MICs over time is more important . With these concepts, a microbiological-pharmacological reading of the antibiogram, taking into account dosage, administration pathway and location of the infectious process among other factors, can be achieved . Most working groups, either national or from abroad, have considered both pharmacokinetics and pharmacodynamics in interpretative reading of the antibiogram . In all cases, breakpoints for susceptibility and resistance should be corrected on the basis of data from well designed and performed clinical trials . Clinically oriented breakpoints do not necessarily have to be the same as those based on microbiological or epidemiological concepts, but clinical microbiologists should be able to give an appropriate response to the question and to the application based on that response. Enferm Infecc Microbiol Clin, 2002 Oct, 20(8), 384 - 7 {Evolution of the use of antibiotics in a hospital long-term care center in Catalonia}; Vaqueiro M et al.; INTRODUCTION: Advanced age, together with immune system changes, malnutrition, chronic disease, and the institutional environment, all contribute to a higher risk of acquiring infection in the elderly . Antibiotics are widely used in geriatric centers, but often their use is not optimal . MATERIAL AND METHODS: Study carried out during the period 1992-1999 in Centro Sociosanitario Albada (Sabadell, Spain) . Data were taken from the Pharmacy Department's unidose registry . We determined the most frequently used antibiotics, the hospital units with highest consumption, the variation in these factors over time, and related costs . RESULTS: A progressive increase in overall antibiotic consumption was observed during the first 5 years of the study with subsequent stabilization . The units showing highest consumption were the Moderate and Highly-Dependent Chronic Unit, the Palliative Care Unit and the Convalescence and Rehabilitation Unit, with significant increases in the Palliative Care Unit in the last two years of the study . Amoxicillin-clavulanate, ciprofloxacin and norfloxacin were the most extensively used antibiotics . Cost increases were seen in the last three years despite the stabilization of antibiotic use . CONCLUSION: We observed a change in the consumption and profile of the antimicrobial agents used in our setting, probably related to changes in the population, increases in parenteral treatment and changes in the criteria for treatment of terminal patients . The establishment of controls for antibiotic use in long-term care centers would lead to improvements in the quality of the care provided. Infect Dis Clin North Am, 2002 Sep, 16(3), 681 - 95 Bone and joint infections in injection drug users; Kak V et al.; Skeletal infections in injection drug users have an insidious onset, present with indolent symptoms, and often occur in unusual locations . Unless physicians are familiar with the disease entities unique to the injection drug user, the diagnosis is frequently delayed . Systemic signs of infection are often lacking . The organisms causing the infection represent a wide spectrum; hence, empiric therapy is not generally recommended . Plain-film radiographs are of little help for early diagnosis . Imaging studies, especially radionucleotide studies and CT or MR imaging scans, can help localize the site of infection . For etiologic diagnosis of these infections, bone biopsy or needle aspiration of the involved bone or joint is required . The choice of antibiotic agent should be based on culture results and the antimicrobial susceptibility of the causative organism . Treatment may also involve surgical drainage or debridement of affected structures . Failure to manage acute bone and joint infection aggressively inevitably leads to chronic, often incurable, infection . Successful therapy requires a team approach including the internist and consultants from orthopedic surgery, infectious diseases, and substance abuse counselors. Minerva Chir, 2002 Oct, 57(5), 543 - 60 The current status and future outlook of intestinal transplantation; Abu-Elmagd K et al.; Recently, the clinical reality of intestinal, combined liver-intestinal, and multivisceral transplantation qualified the procedure by the American Health Care Financing Administration (HCFA) as the standard of care for patients with irreversible intestinal failure . The decision was supported by a decade of clinical experience with cumulative improvement in survival . Prior to tacrolimus, the worldwide experience was plagued with uncontrolled rejection, graft versus host disease (GVHD), and fatal infection . These undefeated barriers stemmed from the large gut lymphoid mass and heavy microbial load contained in the intestinal lumen with the absence of an effective immunosuppressive and antimicrobial therapy . With the emerge of small bowel and multivisceral transplantation in 1990, multiple factors, in addition to the clinical introduction of tacrolimus, have sustained and increased these efforts including evolution in surgical techniques and improvements in postoperative care . The most valuable achievement, however, has been the effective control of rejection and treatment of life threatening opportunistic infections . This chapter outlines the common current practice, surgical techniques and postoperative management of the three different types of intestinal transplantation . In addition, new strategies to overcome some of the current immunologic and biologic challenges are defined with the aim of raising the level of such a creative surgery to be a better therapy for TPN dependent patients. Ann Plast Surg, 2002 Oct, 49(4), 410 - 3 Pyoderma gangrenosum after reduction mammaplasty in an otherwise healthy patient; Lifchez SD et al.; Pyoderma gangrenosum (PG) is a rare postoperative complication of plastic surgery of the breast . Initial signs and symptoms resemble those of infection, and antimicrobial therapy is usually initiated and fails before considering PG as a diagnosis . Therapy consists of immune modulators, and use of corticosteroids is frequent, as is local wound care . Sufficiently small wounds are allowed to heal secondarily, but larger wounds require coverage with either skin grafts or flaps . Long-term (1 year or more) postoperative surveillance is necessary because late failure of the graft or flap can occur. Sex Transm Dis, 2002 Oct, 29(10), 559 - 67 The etiology and management of genital ulcers in the Dominican Republic and Peru; Sanchez J et al.; BACKGROUND: Clinical diagnosis of genital ulcers is difficult, and diagnostic tests are least available in settings where rates of disease are highest . The World Health Organization (WHO) has developed protocols for the syndromic management of genital ulcers in resource-poor settings . However, because risk factors, patterns and causes of disease, and antimicrobial susceptibilities differ from region to region and over time, they must be adapted to local situations . GOAL: The goal of this study was to determine etiologic factors, evaluate syndromic management, and compare polymerase chain reaction (PCR) testing with other diagnostic alternatives for genital ulcers among patients attending sexually transmitted disease clinics in the Dominican Republic and Peru . STUDY DESIGN: Eighty-one men with genital ulcers in the Dominican Republic and 63 in Peru underwent identical interviews and identical multiplex PCR (M-PCR) tests of genital lesion specimens for etiologic diagnoses . Algorithms for managing genital ulcers were developed . RESULTS: In the Dominican Republic, 5% were M-PCR-positive for, 26% for, and 43% for herpes simplex virus (HSV); in Peru, 10%, 5%, and 43%, respectively, were positive . The WHO algorithm for treating syphilis and chancroid had a sensitivity of 100%, a positive predictive value (PPV) of 24%, and an overtreatment rate of 76% . A modified algorithm for treating only those without vesicular lesions had 88% sensitivity and a 27% PPV, and the overtreatment rate was reduced to 58% . CONCLUSION: HSV caused 43% of genital ulcers in these populations . The modified algorithm had lower sensitivity but a reduced overtreatment rate . M-PCR testing was more sensitive than standard tests and more specific and sensitive than clinical diagnosis. Mini Rev Med Chem, 2002 Aug, 2(4), 331 - 42 How do channel- and pore-forming helical peptides interact with lipid membranes and how does this account for their antimicrobial activity? Duclohier H. Animals and plants defend themselves against pathogenic micro-organisms by the rapid mobilization of polycationic helical amphipathic peptides . Interactions with membranes induce optimal orientation and mutual structural changes, allowing for example to form transbilayer ion channels or pores whose properties are compared in this review . Physicochemical studies of peptide-lipid interactions provide attractive approaches for drug design. Protein Pept Lett, 2002 Oct, 9(5), 395 - 402 Antibiotic activity of reversed peptides of alpha-helical antimicrobial peptide, P18; Lee SH et al.; P18 (KWKLFKKIPKFLHLAKKF-NH(2)), an a-helical antimicrobial peptide designed from cecropin Amagainin 2 hybrid, was known to have potent antimicrobial activity against bacteria as well as fungi without hemolytic activity . To find the peptides comparable or superior to the antimicrobial activity of P18, the two reversed peptides (Rev-1 and Rev-2) of P18 were designed and synthesized . These peptides were found to have similar antimicrobial activity against bacterial and fungal cells without hemolytic activity as compared with P18 . Furthermore, a reversed peptide, Rev-2 was shown to have a two-fold higher activity in killing some bacterial cells than P18 . Therefore, these results suggested that Rev-2 peptide seems to be an excellent candidate for developing novel peptide antibiotics. Curr Protein Pept Sci, 2002 Feb, 3(1), 121 - 31 Inhibitory mechanisms of antibiotics targeting elongation factor Tu; Hogg T et al.; Since the pioneering discovery of the inhibitory effects of kirromycin on bacterial elongation factor Tu (EF-Tu) more than 25 years ago {1}, a great wealth of biological data has accumulated concerning protein biosynthesis inhibitors specific for EF-Tu . With the subsequent discovery of over two dozen naturally occurring EF-Tu inhibitors belonging to four different subclasses, EF-Tu has blossomed into an appealing antimicrobial target for rational drug discovery efforts . Very recently, independent crystal structure determinations of EF-Tu in complex with two potent antibiotics, aurodox and GE2270A, have provided structural explanations for the mode of action of these two compounds, and have set the foundation for the design of inhibitors with higher bioavailability, broader spectra, and greater efficacy. Biochemistry, 2002 Oct 15, 41(41), 12359 - 68 Solution and micelle-bound structures of tachyplesin I and its active aromatic linear derivatives; Laederach A et al.; Tachyplesin I is a 17-residue peptide isolated from the horseshoe crab, Tachypleus tridentatus.It has high antimicrobial activity and adopts a beta-hairpin conformation in solution stabilized by two cross-strand disulfide bonds . We report an NMR structural investigation of wild-type tachyplesin I and three linear derivatives (denoted TPY4, TPF4, and TPA4 in which the bridging cysteine residues are uniformly replaced with tyrosine, phenylalanine, and alanine, respectively) . The three-dimensional aqueous solution structures of the wild type and the active variant TPY4 reveal very similar beta-hairpin conformations . In contrast, the inactive variant TPA4 is unstructured in solution . The arrangement of the tyrosine side chains in the TPY4 structure suggests that the beta-hairpin is stabilized by aromatic ring stacking interactions . This is supported by experiments in which the beta-hairpin structure of TPF4 is disrupted by the addition of phenol, but not by the addition of an equimolar amount of cyclohexanol . We have also determined the structures of wild-type tachyplesin I and TPY4 in the presence of dodecylphosphocholine micelles . Both peptides undergo significant conformational rearrangement upon micelle association . Analysis of the micelle-associated peptide structures shows an increased level of exposure of specific hydrophobic side chains and an increased hydrophobic integy moment . Comparison of the structures in micelle and aqueous solution for both wild-type tachyplesin I and TPY4 reveals two requirements for high antimicrobial activity: a beta-hairpin fold in solution and the ability to rearrange critical side chain residues upon membrane association. Plant Mol Biol, 2002 Oct, 50(3), 441 - 52 Over-expression of a seed specific hevein-like antimicrobial peptide from Pharbitis nil enhances resistance to a fungal pathogen in transgenic tobacco plants; Koo JC et al.; Two hevein-like peptides from the seed of Pharbitis nil, designated Pharbitis nil antimicrobial peptide 1 (Pn-AMP1) and Pn-AMP2, had been purified previously . Both exhibit potent in vitro antifungal activity against a broad spectrum of phytopathogenic fungi . We now report the isolation of two cDNA clones, designated pnAMP-h1 and pnAMP-h2, and the corresponding genomic clones encoding these proteins from mature seeds of P . nil . Comparison of the deduced amino acid sequence to that of the mature protein suggests that the peptides are produced as a prepropeptide consisting of an N-terminal signal peptide, the mature protein and C-terminal domains . The transcripts of the two genes are accumulated seed--specifically, and the maximum transcripts are observed in the mid-to-late stage of seed development . Constitutive over-expression of the pnAMP-h2 cDNA in transgenic tobacco under the control of the cauliflower mosaic virus 35S promoter conferred enhanced resistance against the oomycete Phytophthora parasitica, the causal agent of black shank disease . Thus the Pn-AMPs may play a role in the protection of seeds and may be useful as a novel gene source to engineer plants resistant to fungal pathogens. Dent Update, 2002 Sep, 29(7), 325 - 30 Polypharmacy and dentistry: I . Introduction and interactions with local anaesthetics and sedative drugs; Meechan JG; This series of two papers considers the effects of drug interactions in dentistry . In this first paper, the principles of drug interactions will be described . In addition, interactions with drugs used in local anaesthesia and sedation will be discussed . The second paper will concentrate on interactions with analgesics and antimicrobials prescribed in dental practice. Ther Umsch, 2002 Sep, 59(9), 475 - 9 {Genital infections in prepubertal girls}; Navratil F; Vulvitis and vulvovaginitis are the most common gynecologic complaint in prepubertal girls . The frequently observed therapeutic failures are due mainly to lack of knowledge of the characteristics of this age group, of age and development in appropriate diagnostic procedures and of therapeutic measures similar to those in the adult female patient . Prepubertal girls are anatomically, physiologically and behaviorally at relative risk for vulvovaginitis . Symptoms include pruritus, genital pain, "vulvar dysuria" and discharge . History taking and a general pediatric examination are mandatory, thereafter an age appropriate careful anogenital examination should follow . It requires time, patience and knowledge of the different non traumatizing examination techniques . The findings in girls with vulvovaginitis are variable and erythema, excoriations and discharge can be found . The genital inspection and the use of microscopy and microbiologic studies are helpful in planning an appropriate therapy . The majority of vulvovaginal infections in children are nonspecific but they can also be caused by specific organisms and are mostly bacterial . Yeast infections are not found in otherwise healthy prepubertal girls . The therapeutic approach consists of improved anogenital hygiene, sitz/tub baths and use of non irritating soaps . However if an abnormal population of bacteria is present antimicrobial therapy should be considered . Reassurance and a review of preventive methods are crucial in the management of girls with vulvovaginitis. Health Econ, 2002 Oct, 11(7), 637 - 47 Superbugs II: how should economic evaluation be conducted for interventions which aim to contain antimicrobial resistance? Coast J, Smith R, Karcher AM, Wilton P, Millar M. To date, there has been little examination of the problems associated with conducting economic evaluation for interventions designed to contain antimicrobial resistance . There are two quite different types of intervention aimed at containing antimicrobial resistance: interventions which are designed to avoid the emergence of resistant organisms; and interventions that are designed to avoid the transmission of resistance organisms . Four aspects of economic evaluation where the ease of assessment might be expected to differ across evaluations for these different types of intervention are examined: problems associated with the identification of diffuse impacts, problems associated with comparing current and future impacts, problems associated with uncertainty, and problems associated with difficulties in measurement and valuation . The paper suggests that it may be much easier to conduct rigorous economic evaluations for interventions designed to avoid transmission of resistance, than for those intended to avoid emergence . Unfortunately, the transmission policies, which are likely to be the easiest to evaluate, are not likely to produce an optimal long-term outcome given the apparent irreversibility of much resistance and the potentially severe harms which could be imposed as a result . Given the desirability of avoiding a scenario where, in the evidence-based medicine culture, the most rigorously evaluated policies are followed even though they may be less important, there is the need to consider carefully what, and how, economic evaluation should be conducted in the area of antimicrobial resistance . It is suggested that research should focus on the use of modelling as a means of evaluating optimal policy responses and on trying to resolve some of the difficulties associated with measurement and valuation . J Pediatr Hematol Oncol, 2002 Oct, 24(7), 558 - 60 Nontuberculous mycobacterial infections in pediatric acute leukemia; Suryanarayan K et al.; We report on 3 children undergoing treatment for acute lymphoblastic leukemia (ALL), who developed systemic nontuberculous mycobacterial (NTM) infections . All 3 patients were treated successfully with 5 months or less of antimicrobial therapy and completed their chemotherapy with no further recurrence of their NTM infection . NTM infections in some children with ALL may be successfully treated with antimicrobial agents without necessarily compromising the ALL treatment . The optimal duration of therapy for NTM remains unclear, but may be shorter than previously reported. Laryngoscope, 2002 Oct, 112(10), 1758 - 61 Outpatient intravenous antibiotics for chronic rhinosinusitis; Gross ND et al.; OBJECTIVE/HYPOTHESIS: Peripherally inserted central catheter (PICC) lines have greatly facilitated the use of intravenous antibiotics in outpatient medical practice . Otolaryngologic applications for home intravenous therapy through PICC lines have not been well characterized to date . The purpose of the study is to describe indications and complications related to outpatient intravenous antibiotic therapy in patients with chronic rhinosinusitis . STUDY DESIGN: Retrospective cohort study . METHODS: Chart review of patients with chronic rhinosinusitis who were treated at an academic rhinology practice with outpatient intravenous antibiotics over a 3-year period.RESULTS Fourteen patients receiving, in all, 16 courses of intravenous antibiotic therapy through PICC line were identified . The average patient age was 51 years (age range, 36-74 y) . The primary indication for intravenous antibiotic use was the treatment of resistant pathogens (50%) . The most common organisms treated were, and . Other indications included gastrointestinal intolerance of oral antibiotics and extranasal complications of sinusitis . Eighty-eight percent of patients (14 of 16) were able to complete the entire prescribed course of therapy . Three (19%) catheter-related complications occurred, including thrombophlebitis and deep venous thrombosis . All three complications required removal of the PICC line; one of these patients underwent successful reinsertion of a second catheter and completion of treatment . CONCLUSIONS: Peripherally inserted central catheter line delivery of home intravenous antibiotics can be a well-tolerated adjunct to surgery in the outpatient treatment of chronic rhinosinusitis . Resistant infections, intolerance to oral antimicrobials, and extranasal complications of sinusitis are indications for PICC line therapy . Catheter-related complications can be significant and must be considered in patient selection. Toxicol Lett, 2002 Nov 15, 136(1), 77 - 84 Studies on cytotoxic and genotoxic effects of N-hydroxypyridine-2-thione (Omadine) in L5178Y mouse lymphoma cells; Moller M et al.; The cytotoxicity and genotoxicity of the antifungal and antimicrobial agent Omadine, i.e . N-hydroxypyridine-2-thione (HOPT), has been investigated in L5178Y mouse lymphoma cells in the dark and under UVA irradiation . Omadine inhibits cell growth and induces micronuclei at concentrations >0.5 microM in the absence of light . At a 0.5-microM concentration, an UVA-dose-dependent induction of micronuclei is observed, conditions at which the cytotoxicity and genotoxicity in the dark is negligible . The photogenotoxicity is not accompanied by cytotoxicity . Control experiments with the radical scavengers GSH and GSHOEt implicate the involvement of hydroxyl radicals in the photogenotoxicity of Omadine. Neuropharmacology, 2002 Sep, 43(4), 778 - 87 Characterization of the interaction between a novel convulsant agent, norbiphen, and GABA(A) and other ligand-gated ion channels; Halliwell RF et al.; A hybrid molecule composed of the antimicrobial, norfloxacin, linked to the non-steroidal anti-inflammatory drug (NSAID), biphenylacetic acid, which we have termed norbiphen, is a lethal convulsant in vivo and an antagonist of rodent GABA(A) receptors in vitro . In the present study, the selectivity, molecular site(s) and mechanism of action of this novel convulsant were investigated using electrophysiological techniques . Sub-maximal GABA-evoked currents recorded from rodent hippocampal neurons were reversibly inhibited by norbiphen (1 microM) to 5+/-2% of control whereas glutamate, NMDA and glycine activated responses were little or unaffected . Sub-maximal GABA-evoked currents recorded from oocytes expressing recombinant human alpha1beta2gamma2s or alpha1beta2 GABA(A) receptors were also reversibly inhibited by norbiphen (1-1000 nM) with an IC(50) (+/-s.e.m.) of 5.7+/-1 and 8.8+/-1 nM, respectively . Similarly, GABA currents recorded from alpha1beta1gamma2s, alpha1beta1 and beta2gamma2s receptors were inhibited with IC(50)s of 16.1+/-1, 18.8+/-1 and 4.2+/-1 nM, respectively . In contrast, norbiphen (100 nM) had little or no effect at rho1 GABA(C) homomers . At alpha1beta2gamma2s receptors, norbiphen had no affect on the GABA reversal potential, and inhibition was not voltage-dependent, suggesting that this compound does not act at the ion channel . The GABA concentration response curve was shifted in a competitive-like fashion by norbiphen (10-300 nM) and a Schild analysis of these data yielded a slope of 0.94+/-0.1 and a pA(2) of 7.77 . Our data reveal a novel, selective and highly potent antagonist of GABA(A) receptors . Norbiphen should be a valuable agent in future studies of this receptor complex. J Periodontal Res, 2002 Oct, 37(5), 389 - 98 Selection of antimicrobial agents in periodontal therapy; Slots J; BACKGROUND: The recognition over the past 3 decades of microbial specificity in periodontitis has afforded dental practitioners the ability to prevent and treat the disease with a variety of antimicrobial drugs . These include systemic antibiotics, topical antibiotics and topical antiseptics . RESULTS: Systemic antibiotic therapy can be essential in eliminating pathogenic bacteria that invade gingival tissue and in helping control periodontal pathogens residing in various domains of the mouth from where they may translocate to periodontal sites . Frequently used periodontal combination antibiotic therapies are metronidazole-amoxicillin (250-375 mg of each 3 x daily for 8 days) and metronidazole-ciprofloxacin (500 mg of each 2 x daily for 8 days) . Microbiological analysis helps determine the optimal antibiotic therapy and effectiveness of treatment . Topical antibiotics that are commercially available as controlled release devices suffer from several potential problems, including insufficient spectrum of antimicrobial activity in some periodontal polymicrobial infections, risks of producing an antibiotic resistant microbiota, and high acquisition costs . Topical antiseptics of relevance in periodontal treatment include 10% povidone-iodine placed subgingivally by a syringe for 5 min, and 0.1% sodium hypochlorite solution applied subgingivally by patients using an irrigation device . CLINICAL IMPLICATIONS: The present paper recommends periodontal treatment that includes a battery of professionally and patient-administered antimicrobial agents (properly prescribed systemic antibiotics, povidone-iodine and sodium hypochlorite subgingival irrigants, and chlorhexidine mouthrinse) . Available chemotherapeutics can provide effective, safe, practical and affordable means of controlling subgingival colonization of periodontal pathogens and various types of periodontal disease. J Pept Res, 2002 Oct, 60(4), 198 - 214 Exploring relationships between mimic configuration, peptide conformation and biological activity in indolizidin-2-one amino acid analogs of gramicidin S; Roy S et al.; Indolizidin-2-one amino acids (I2aas, 6S- and 6R-1) possessing 6S- and 6R-ring-fusion stereochemistry were introduced into the antimicrobial peptide gramicidin S (GS) to explore the relationships between configuration, peptide conformation and biological activity . Solution-phase and solid-phase techniques were used to synthesize three analogs with I2aa residues in place of the d-Phe-Pro residues at the turn regions of GS: {(6S)-I2aa4-5,4'-5'}GS (2), {Lys2,2',(6S)-I2aa4-5,4'-5'}GS (3) and {(6R)-I2aa4-5,4'-5'}GS (4) . Although conformational analysis of {I2aa4-5,4'-5'}GS analogs 2-4 indicated that both ring-fusion stereoisomers of I2aa gave peptides with CD and NMR spectral data characteristic of GS, the (6S)-I2aa analogs 2 and 3 exhibited more intense CD curve shapes, as well as greater numbers of nonsequential NOE between opposing Val and Leu residues, relative to the (6R)-I2aa analog 4, suggesting a greater propensity for the (6S)-diastereomer to adopt the beta-turn/antiparallel beta-pleated sheet conformation . In measurements of antibacterial and antifungal activity, the (6S)-I2aa analog 2 exhibited significantly better potency than the (6R)-I2aa diastereomer 4 . Relative to GS, {(6S)-I2aa4-5,4'-5'}GS (2) exhibited usually 1/2 to 1/4 antimicrobial activity as well as 1/4 hemolytic activity . In certain cases, antimicrobial and hemolytic activities of GS were shown to be dissociated through modification at the peptide turn regions with the (6S)-I2aa diastereomer . The synthesis and evaluation of GS analogs 2-4 has furnished new insight into the importance of ring-fusion stereochemistry for turn mimicry by indolizidin-2-one amino acids as well as novel antimicrobial peptides. Microbiol Immunol, 2002, 46(8), 571 - 3 Menadione-catalyzed luminol chemiluminescent assay for viability of Mycobacterium bovis; Yamashoji S; Stable luminol chemiluminescence was observed 10 min after the addition of menadione to a suspension of Mycobacterium bovis homogenized in Middlebrook 7H9 broth base including OADC enrichment . The chemiluminescence intensity was proportional to the absorbance of the bacterial suspension at 600 nm in a range of 0.005 to 0.15 . Luminol chemiluminescence disappeared after 10 min incubation of M . bovis at over 60% of ethanol or 4 days of cultivation of M . bovis in the presence of 40 microg/ml of streptomycin . The bacterium showing the disappearance of chemiluminescence could not grow after being washed, suggesting that the inhibition concentration of the antimicrobials can be estimated on the basis of the disappearance of chemiluminescence . Menadione-catalyzed luminol chemiluminescent assay was rapid and sensitive in comparison to turbidimetry, tetrazolium (WST-8) reduction assay, and the assay using the Mycobacteria growth indicator tube (MGIT). Microb Drug Resist, 2002 Fall, 8(3), 187 - 92 Clonal spread of resistant pneumococci despite diminished antimicrobial use; Arason VA et al.; The effects of community-wide interventions to reduce resistance rates are poorly understood . This study evaluated the effect of reduced antimicrobial usage on the spread of penicillin-nonsusceptible pneumococci (PNSP) in four communities in Iceland . The study was performed after interventions to reduce antimicrobial usage and compared to an identical study performed 5 years before . A randomized sample of 953 children was chosen from all 2,900 1- to 6-year-old children living in four well-defined communities . The main outcome measures were nasopharyngeal carriage of PNSP and individual and community use of antimicrobials . Pneumococci were carried by 51.7% of the 743 children enrolled, and 8.1% of the pneumococci were PNSP as opposed to 8.5% in the previous study . The antimicrobial use of participants had been reduced from 1.5 to 1.1 courses/year and the overall use among children <7 years old living in the study areas from 13.6 to 11.1 defined daily dosages/1000 children per day . The prevalence of PNSP increased in the two areas furthest away from the capital area despite reduced consumption . The major risk factors for carriage of PNSP remained the same . Interventions can be effective in reducing antimicrobial use . Pandemic multiresistant clones can also spread fast in small communities with low antimicrobial use, where their appearance may be delayed compared to highly populated urban areas . Clonal spread and herd immunity are important factors to be considered in the evaluation of intervention effects. Can J Gastroenterol, 2002 Sep, 16(9), 611 - 4 Motion--Helicobacter pylori worsens GERD: arguments for the motion; O'Morain CA et al.; There are several reasons for eradicating Helicobacter pylori in patients with chronic gastroesophageal reflux disease (GERD) . Perhaps the most compelling is the evidence that chronic acid suppression therapy can lead to the development of atrophic gastritis, a premalignant condition, in patients with H pylori infection . Epidemiological data that suggest that H pylori is less prevalent in GERD patients than in control subjects may be susceptible to publication bias, and confounding social and environmental factors may also be involved . Although it has been thought that eradication of the organism might lead to increased esophageal acid exposure, this has not been demonstrated in practice . Studies that appeared to show that GERD could be provoked by antimicrobial therapy of duodenal ulcers also have methodological weaknesses . Underlying GERD symptoms might be unmasked after withdrawal of acid-suppression therapy, for reasons that are unrelated to H pylori . In fact, eradication of the organism has been shown to decrease heartburn in patients with peptic ulcer disease . When H pylori is successfully eradicated in patients with GERD, relapse rates are not increased, and the disease-free interval seems to be prolonged . Eradication of the organism is a wise policy in patients who face long term acid-suppression therapy for GERD. Eur J Gastroenterol Hepatol, 2002 Oct, 14(10), 1093 - 100 Five minute endoscopic urea breath test with 25 mg of (13)C-urea in the management of Helicobacter pylori infection; Isomoto H et al.; OBJECTIVE: The endoscopic (13)C-urea breath test ((13)C-EUBT), which combines the urea breath test (UBT) with endoscopy, provides high accuracy for the detection of Helicobacter pylori . This study was conducted to determine whether the (13)C-EUBT using low doses of urea and short sampling times could preserve accuracy in the management of H . pylori infection . METHODS: Three hundred and twenty-five patients were randomized to receive the EUBT with 100, 50 or 25 mg of (13)C-urea by endoscopic spraying . The breath samples collected at 5, 10 and 20 min were analysed using an isotope selected non-dispersive infrared spectrometer . H . pylori infection was assessed by the rapid urease test and histology . In each sampling schedule and protocol, cut-off values were calculated by a receiver operating characteristic curve . We applied the EUBT with 25 mg of (13)C-urea at 5 min to the assessment of eradication in 135 patients who had received the antimicrobial treatment or to the detection of the organism in 61 patients with previous partial gastrectomy . RESULTS: Based on histology and the urease test, patients who had discordant results were excluded from the analysis . Using 100 mg of urea, the sensitivity and specificity of the test were both 100% at 10 and 20 min, and the sensitivity and specificity at 5 min were best with 98.6% and 100%, respectively . With 50 mg, they were both 100% at 20 min, and the best combination of sensitivity and specificity at 5 and 10 min was 97.3-96.6% and 97.3-100%, respectively . Even with 25 mg, the sensitivity and specificity were both 100.0% at 20 min, and at the 5 min and 10 min time point, the EUBT yielded a sensitivity of 98.7% and a specificity of 100% . There was a significant positive correlation between the test values of the 5 min (13)C-EUBT with 25 mg of test urea and those of the conventional UBT . The 5 min EUBT with (13)C-urea offered high accuracy in the assessment of H . pylori eradication, with the sensitivity and specificity being 100% and 96.4%, respectively . In patients with previous gastrectomy, the EUBT provided acceptable accuracy (a sensitivity of 96.4% and a specificity of 97.0%) . CONCLUSIONS: Our results indicate that the (13)C-EUBT is an accurate method for detecting H . pylori infection . The EUBT using only 25 mg of (13)C-urea at the early (5 min) time point has satisfactory diagnostic efficacy in pre- and post-eradication treatment settings, providing a less expensive and more rapid way of performing the test . The EUBT may be a reliable method of assessing H . pylori status in the remnant stomach. Farmaco, 2002 Aug, 57(8), 631 - 9 Synthesis of variously substituted 1,8-naphthyridine derivatives and evaluation of their antimycobacterial activity; Badawneh M et al.; A series of 1,8-naphthyridine derivatives variously substituted in the 2, 3, 4 and 7 positions were synthesized for in vitro evaluation of antimycobacterial activity in accordance with an international program with the tuberculosis antimicrobial acquisition and coordinating facility (TAACF) . Several compounds 4, 8, 12, 14, 19, 29 and 30, when tested at a concentration of 6.25 microg/ml against Mycobacterium tuberculosis H37Rv, showed an interesting activity with % inhibition in the range 38-96% . The most effective substituent in position 2, 4 or 7 of the 1,8-naphthyridine nucleus seem to be the piperidinyl group. Clin Orthop, 2002 Oct, (403), 80 - 6 Outpatient intravenous antimicrobial therapy for the practicing orthopaedic surgeon; Osmon DR et al.; The medical treatment of many musculoskeletal infections requires prolonged intravenous antimicrobial therapy, much of which is administered outside the hospital under the guidance of an infectious disease specialist . The essential elements of patient selection, antimicrobial administration, and clinical and laboratory monitoring of outpatient intravenous antimicrobial therapy for adult patients with musculoskeletal infection is described in the current study . Orthopaedists who treat patients with musculoskeletal infection should know the essential details of outpatient intravenous antimicrobial therapy to optimize the medical and surgical treatment of patients with these infections. Clin Orthop, 2002 Oct, (403), 49 - 53 Antimicrobial release kinetics from polymethylmethacrylate in a novel continuous flow chamber; Perry AC et al.; Polymethylmethacrylate is used for local delivery of antimicrobials in the treatment of musculoskeletal infections . A novel continuous flow chamber system was designed to measure in vitro antimicrobial release . Three-millimeter beads containing amikacin, gentamicin, tobramycin, or vancomycin {concentration of 7.5% (weight per weight)} were placed individually in a continuous flow chamber with a total volume of 1 mL Kreb's Ringer buffer flowing at 1 mL/hour . Effluent was sampled hourly for 24 hours and then every 2 hours up to 48 hours; antimicrobial concentrations were measured in triplicate by bioassay . The mean peak concentrations were 40.9, 30.1, 30.0, and 19.1 microg/mL; the mean areas under the concentration time curves (Time 0 to infinity) were 263, 327, 110, and 180 hours x microg/mL of antibiotic; and the mean percentages of initial amount of antimicrobial released were 11.7%, 14.5%, 6.6%, and 10.9% for tobramycin, gentamicin, amikacin, and vancomycin, respectively . The results for each polymethylmethacrylate-antimicrobial agent combination were reproducible . In contrast to other in vitro elution systems, this novel system operates under the premise that there is dynamic flow surrounding polymethylmethacrylate in vivo and permits rapid in vitro comparison of the relative release of antimicrobial agents from polymethylmethacrylate. Proc Natl Acad Sci U S A, 2002 Oct 15, 99(21), 13705 - 10 Epub 2002 Oct 01. Overexpression of a pattern-recognition receptor, peptidoglycan-recognition protein-LE, activates imd/relish-mediated antibacterial defense and the prophenoloxidase cascade in Drosophila larvae; Takehana A et al.; In Drosophila, microbial infection activates an antimicrobial defense system involving the activation of proteolytic cascades in the hemolymph and intracellular signaling pathways, the immune deficiency (imd) and Toll pathways, in immune-responsive tissues . The mechanisms for microbial recognition are largely unknown . We report that, in larvae, the imd-mediated antibacterial defense is activated by peptidoglycan-recognition protein (PGRP)-LE, a PGRP-family member in Drosophila . Consistent with this, PGRP-LE binds to the diaminopimelic acid-type peptidoglycan, a cell-wall component of the bacteria capable of activating the imd pathway, but not to the lysine-type peptidoglycan . Moreover, PGRP-LE activates the prophenoloxidase cascade, a proteolytic cascade in the hemolymph . Therefore, PGRP-LE acts as a pattern-recognition receptor to the diaminopimelic acid-type peptidoglycan and activates both the proteolytic cascade and intracellular signaling in Drosophila immunity. Pediatrics, 2002 Oct, 110(4), 805 - 14 The future of pneumococcal conjugate vaccines for prevention of pneumococcal diseases in infants and children; Pelton SI et al.; Seven-valent pneumococcal conjugate vaccine (PCV7) was licensed in February 2000 . In June 2000, the Advisory Committee on Immunization Practices and the American Academy of Pediatrics recommended the universal administration of pneumococcal conjugate vaccine for all children 23 months of age and younger and for children 24 to 59 months of age who are at high risk for serious pneumococcal disease . Since then, >23 million doses have been administered in the United States . Postlicensure surveillance of invasive pneumococcal disease (IPD) in the United States from the Active Bacterial Core Surveillance program at the Centers for Disease Control and Prevention and the Northern California Kaiser Permanente Vaccine Study Center has reported a decline in IPD and in pneumococcal disease incidence as a result of vaccine serotypes, respectively . During this period, issues critical to the long-term success of PCV7 have become more relevant: Will PCV7 be as effective in groups of children who are at high risk for IPD as in healthy children? Will nonvaccine types replace vaccine serotypes in the nasopharynx and in disease? Why are the results of the clinical trials different for IPD and for acute otitis media? How many doses of PCV7 and what concentrations of antibody are necessary for protection? Will universal administration of PCV7 to children younger than 2 years reduce antimicrobial drug resistance and alter prescribing patterns of physicians for febrile infants? Have there been unanticipated adverse events or benefits observed? The purpose of this report is to review the current data available to address these questions and to identify gaps that will require additional knowledge to determine the ultimate value of pneumococcal conjugate vaccines in reducing the burden of pneumococcal disease in infants and children. Acad Emerg Med, 2002 Oct, 9(10), 977 - 82 The effects of a high-potency topical steroid on cutaneous healing of burns in pigs; Singer AJ et al.; OBJECTIVE: Burns are dynamic injuries that tend to progress over the course of several days . Steroids inhibit the formation of vasoconstrictive prostanoids that may contribute to this progression of injury . The authors hypothesized that adding topical steroids to a standard antimicrobial agent would reduce the progression of burns and accelerate reepithelialization without increasing infection rates . METHODS: This was a prospective, blinded, controlled, experimental trial . Forty-eight standardized second-degree burns were created by applying an aluminum bar preheated to 80 degrees C to the flanks of isoflurane-anesthetized young pigs for 20 seconds . Three equal sets of 16 burns were randomly treated with silver sulfadiazine cream (SSD), clobetasol propionate 0.05% (CP), or both (SSD+CP) . Daily dressing changes were performed for 14 days . Full-thickness biopsies were taken after injury and at one, two, seven, ten, and 14 days for blinded histopathological evaluation using hematoxylin and eosin (H&E) staining . The primary outcome was the % reepithelialization (REP) calculated by dividing the length of the neoepidermis by the section's total length (interobserver correlation = 0.99) . Depth of injury was measured for each dermal element (collagen; epithelial, mesenchymal, and vascular cells; and vessel thrombosis) . Comparisons across groups were performed using one-way analysis of variance (ANOVA) . A repeated-measures ANOVA was used to compare injury depths over time . This study had 80% power to detect a 33-percentage point difference in REP across groups (two-tailed alpha = 0.05) . RESULTS: Pretreatment burn depths were similar across groups . While burn depth changed over time, there was no difference between the groups in burn injury progression . There was no difference across the groups in REP or infection rates at all times . CONCLUSIONS: Addition of a potent topical steroid to standard antimicrobial topical agents does not reduce burn depth or accelerate reepithelialization after burns. Lett Appl Microbiol, 2002, 35(4), 285 - 90 Study on the mechanisms of the antibacterial action of some plant alpha,beta-unsaturated aldehydes; Trombetta D et al.; AIMS: In this paper the mechanisms involved in the antibacterial effect of six 2E-alkenals {(E)-2-hexenal, (E)-2-eptenal, (E)-2-octenal, (E)-2-nonenal, (E)-2-decenal and (E,E)-2,4-decadienal} were investigated . METHODS AND RESULTS: We measured the release of carboxyfluorescein (CF) trapped in liposomes of phosphatidylcholine (PC) following exposure to the aldehydes mentioned above, in comparison with that elicited by hexanal and nonanal; the modifications of the thermotropic behaviour of liposomes of dimyristoylphosphatidylcholine (DMPC) induced by (E,E)-2,4-decadienal (the aldehyde endowed with the highest microbicidal activity) were evaluated by means of differential scanning calorimetry . With the exception of hexanal, all aldehydes tested caused rapid CF leakage from PC liposomes . The effectiveness order correlates well with the chain length and the presence of the alpha,beta-double bond . Furthermore (E,E)-2,4-decadienal is able to interact with and cross DMPC bilayers . CONCLUSIONS: The present findings suggest that the 2E-alkenals tested elicit, very likely, a gross perturbation of the lipidic fraction of plasmatic membranes and are able to penetrate into bacterial cells . SIGNIFICANCE AND IMPACT OF THE STUDY: These data represent an interesting background for a rational employment of the plant 2E-alkenals tested as antimicrobial agents. Am J Clin Dermatol, 2002, 3(8), 529 - 34 Topical treatment of pediatric patients with burns: a practical guide; Palmieri TL et al.; Over 440 000 children receive medical attention for burn injuries each year in the US . Burn wound infections are a major source of morbidity and mortality in these patients . Infected wounds not only heal more slowly, but also may lead to systemic infections . The factors that contribute to wound complications are both the size and depth of the wound . Burn depth is usually categorized into first-degree (superficial, involving only the epidermis), second-degree (partial thickness, involving both epidermis and dermis), and third-degree (full thickness, through the epidermis, dermis, and into fat) . Burns that will not heal within 2 weeks are at least second-degree and should generally be referred to a burn surgeon for possible excision and grafting, due to the increased risk of infection and scarring . The burn wound is dynamic . Proper treatment minimizes the extent of the burn injury, whereas improper treatment (lack of proper wound-care, edema formation, lack of resuscitation) may actually increase the size and/or depth of the wound . Topical antimicrobial agents have been shown to decrease wound-related infections and morbidity in burn wounds when used appropriately . The goal of topical antimicrobial therapy is to control microbial colonization, thus preventing development of invasive infections . A wide variety of agents are available for treatment of burn wounds, including ointments, creams, biological and nonbiological dressings . Topical antimicrobials of choice include bacitracin, neomycin, silver sulfadiazine and mafenide. J Med Microbiol, 2002 Sep, 51(9), 786 - 90 Prevalence of resistant Helicobacter pylori isolates in Bulgarian children; Boyanova L et al.; The aim of this study was to assess the primary and combined resistances of Helicobacter pylori isolates obtained from paediatric patients in 2000-2001 to seven antimicrobial agents . Resistance rates of pre-treatment isolates from 115 children were investigated by the limited agar dilution method alone and by the E-test . The cut-off concentrations for resistance were: metronidazole >8 mg/L, clarithromycin and azithromycin >1 mg/L, clindamycin >4 mg/L, amoxicillin >0.5 mg/L, tetracycline >4 mg/L and ciprofloxacin >1 mg/L . Primary resistance rates were: metronidazole 15.8%, clarithromycin 12.4%, azithromycin 14.6%, clindamycin 20.0%, amoxicillin 0%, metronidazole + clarithromycin 4.5%, ciprofloxacin 6.0%, metronidazole + clarithromycin + ciprofloxacin 1.2%, tetracycline 3.1% and metronidazole + ciprofloxacin 1.2% . There were no significant age (1-9 years versus 10-18 years) or gender differences . Prevalence of both macrolide-resistant and intermediately susceptible strains was 21.9% for azithromycin and 15.9% for clarithromycin . Of 18 metronidazole-resistant isolates, 77.8% exhibited a metronidazole MIC > or = 32 mg/L . H . pylori resistance rates to metronidazole, clarithromycin and both agents were relatively low in Bulgarian children . However, resistance was found to all drugs tested except for amoxicillin . The consumption of newer macrolides and tetracyclines could be related to the prevalence of resistance to the corresponding agents . There were no significant differences in primary resistance rates of H . pylori to antimicrobial agents between children and adults except for metronidazole . Multi-drug resistance to newer macrolides, metronidazole and ciprofloxacin in association with a slightly elevated amoxicillin MIC (0.38 mg/L) was detected in one strain. Equine Vet J, 2002 Sep, 34(6), 542 - 8 Chronic pulmonary disease with radiographic interstitial opacity (interstitial pneumonia) in foals; Nout YS et al.; Twelve foals, age 3-9 months, examined at The Ohio State University Veterinary Teaching Hospital between 1995 and 2000 were diagnosed with chronic pulmonary disease associated with marked interstitial opacity on radiographic examination . The most characteristic features were a history of respiratory disease of 1-3 months duration, marked clinical signs of respiratory disease, failure to yield a consistent pathogen from tracheobronchial aspirates and a predominantly interstitial pattern on thoracic radiographs . We attributed these signs to chronic interstitial pneumonia . Foals were treated with broad spectrum antimicrobial and corticosteroid drugs . All 12 foals were discharged alive from hospital and, of the 10 available for follow-up, all were disease-free and performing to expectation 5 months to 5 years after discharge . We conclude that chronic interstitial pneumonia, occuring in foals, is associated with a good prognosis and that corticosteroid therapy may be useful in its treatment. Planta Med, 2002 Sep, 68(9), 836 - 8 Chemical analysis and antifungal activity of the essential oil of Calea clematidea; Flach A et al.; The chemical composition of the essential oils of Calea clematidea Baker obtained by hydrodistillation of the leaves and flowers was analysed by GC and GC/MS and the oils were assayed for their antifungal activities . The essential oil of the leaves showed a high content of a new natural epoxy terpenoid, named clemateol (ca . 70 %), with minor amounts of o-vanillin (6.5 %), spathulenol (4.2 %), alpha-terpinene (4.0 %), germacrene B (2.9 %), yomogi alcohol (1.8 %), ( E)-caryophylene (1.7 %), m-cymenene (1.6 %), and alpha-gurjunene (1.5 %), while the essential oil of the flowers was characterized by a higher content of thymol methyl ether (ca . 80 %), with minor amounts of clemateol (4.8 %) and o-cymene (4.7 %) . The antimicrobial activity of the oils was also evaluated against dermatophytes for their possible use in pharmaceutical preparations for topical applications . The oil of the leaves (MIC > 3.57 mg/ml), clemateol (MIC > 1.52 mg/ml), and the alcohol 2 (MIC > 2.82 mg/ml) showed a moderate antifungal activity against Trichophyton tonsurans, Trichophyton rubrum, Trichophyton menthagrophytes var . i nterdigitale, Epidermophyton floccosum, Microsporum gypseum, Microsporum canis and Microsporum nanum. Planta Med, 2002 Sep, 68(9), 808 - 12 Antimicrobial terpenoids from the oleoresin of the Peruvian medicinal plant Copaifera paupera; Tincusi BM et al.; Twelve known diterpenes 1 - 11 and 13, and three known sesquiterpenes 14 - 16, along with a new C(20) - C(15) terpenoid 17, with a structure based on an unprecedented skeleton in which a labdane diterpene is linked to a monocyclic sesquiterpene by an ester bridge, were isolated from the oleoresin of the Peruvian medicinal plant Copaifera paupera (Herzog) Dwyer (Leguminosae) . Their structures were elucidated on the basis of spectral analysis, including homo- and heteronuclear correlation NMR experiments (COSY, ROESY, HMQC and HMBC), and by comparison with data in the literature . The leishmanicidal, antimicrobial, cytotoxic, and aldose reductase inhibitory activities were studied . Compounds 1 and 11 showed significant antimicrobial activity (MIC < 10 microg/ml) against Gram-positive bacteria, comparable with cephotaxime used as control . Compound 2 exhibited moderate cytotoxic activity against four cancer cell lines. Curr Opin Crit Care, 2002 Oct, 8(5), 465 - 72 New concepts in sepsis; Sessler CN et al.; An estimated 750,000 cases of severe sepsis occur annually in the United States, and the mortality rate is about 30% . As a condition that disproportionately affects the elderly and is related to invasive and immunosuppressive healthcare, increases in the frequency of sepsis are anticipated . The complex pathophysiology of sepsis encompasses the interplay of pro- and anti-inflammatory mediators, activated circulating and resident inflammatory cells, disrupted coagulation, endothelial activation and injury, vasodilatation and vascular hyporesponsiveness to vasoactive mediators, cardiac dysfunction, and cellular dysoxia . Current management of severe sepsis includes eradication of infection through source control and antimicrobial therapy, aggressive and targeted shock resuscitation that includes fluid administration, correction of anemia, vasopressor support, modest inotropic therapy, infusion of human recombinant activated protein C to selected patients, and compulsive supportive care to manage organ dysfunction and to avoid complications. Curr Opin Crit Care, 2002 Oct, 8(5), 453 - 60 Severe community-acquired pneumonia; Ewig S et al.; Although several pneumonia severity criteria have been firmly established, the exact definition of severe community-acquired pneumonia (CAP) remains elusive . Mortality from CAP remains high, reaching 50% in some series . The particular role of and spp . in severe CAP has been defined more clearly . Microbial diagnosis in the individual patient remains a difficult task . Despite promising new diagnostic tools, concerns about possible mixed origins preclude a change from the currently advocated broad-spectrum approach of antimicrobial treatment . Although there is some evidence that guidelines may optimize outcomes, their role in limiting the spread of resistance has only recently received attention . Finally, although there are promising data on the use of noninvasive positive pressure ventilation to treat pneumonia in patients without chronic obstructive pulmonary disease, its place in the management of acute respiratory failure remains to be defined in randomized studies. Curr Opin Crit Care, 2002 Oct, 8(5), 435 - 40 Optimizing antimicrobial dosing in the critically ill patient; Goldberg J et al.; "We know everything about antibiotics except how much to give," Maxwell Finland once stated . Finally, with the proliferation of pharmacodynamics as a science, we are addressing the question of how much to give . We have moved from an era of more or less arbitrary antimicrobial dosage selection toward one characterized by evidence-based optimal dosing strategies . Optimizing antimicrobial therapy in critically ill patients is more than just the selection of a suitable dose for a particular patient . Optimizing therapy also involves the selection of an appropriate single or combination antibiotic regimen that is active against the suspected or documented pathogens at the site(s) of infection . The regimen should offer the fewest potential adverse events, and the duration of therapy should be the shortest possible so as not to encourage resistance . Dosing of the chosen regimen should reflect variables that are often ignored, such as the patient's weight and age . The new continuous renal replacement therapies are commonly used in the critical care unit and must be considered . Finally, the cost of the regimen should be considered, but not only the cost to purchase the chosen antimicrobial agent but the cost to administer it (, the cost of minibags or syringes, intravenous tubing, saline flushes {all multiplied by the number of times per day the drug is given}), and, most importantly, if the patient fails to respond to therapy, the cost necessary to re-treat the patient to bring about a cure . In this review, we discuss some of the principles required to optimize antimicrobial dosing and recently obtained data regarding its application to the critically ill patient. Curr Opin Crit Care, 2002 Oct, 8(5), 421 - 9 Nosocomial pneumonia: emerging concepts in diagnosis, management, and prophylaxis; Craven DE et al.; Nosocomial pneumonia is a dynamic disease with multiple etiologic agents and a changing natural history . The highest attack rates and mortality occur in patients with ventilator-associated pneumonia . Diagnosis of nosocomial pneumonia is often made by clinical criteria that are sensitive but lack specificity . The use of quantitative endotracheal aspirates or bronchoscopy with bronchoalveolar lavage and protected specimen brush clearly improve diagnostic specificity and outcome in patients who are mechanically ventilated . The rapid spread of multidrug-resistant, spp, and has made initial empiric therapy more difficult . Management principles include the use of techniques for more accurate diagnosis and early antimicrobial therapy with appropriate agents along with careful analysis of culture results, clinical response, and potential complications of pneumonia and therapy . Strategies for prophylaxis are of critical importance for risk reduction, improvement in patient outcome, and reduction of hospital costs. Proc Natl Acad Sci U S A, 2002 Oct 15, 99(21), 13560 - 5 Epub 2002 Sep 30. Solution structure and dynamics of the outer membrane enzyme PagP by NMR; Hwang PM et al.; The bacterial outer membrane enzyme PagP transfers a palmitate chain from a phospholipid to lipid A . In a number of pathogenic Gram-negative bacteria, PagP confers resistance to certain cationic antimicrobial peptides produced during the host innate immune response . The global fold of Escherichia coli PagP was determined in both dodecylphosphocholine and n-octyl-beta-d-glucoside detergent micelles using solution NMR spectroscopy . PagP consists of an eight-stranded anti-parallel beta-barrel preceded by an amphipathic alpha helix . The beta-barrel is well defined, whereas NMR relaxation measurements reveal considerable mobility in the loops connecting individual beta-strands . Three amino acid residues critical for enzymatic activity localize to extracellular loops near the membrane interface, positioning them optimally to interact with the polar headgroups of lipid A . Hence, the active site of PagP is situated on the outer surface of the outer membrane . Because the phospholipids that donate palmitate in the enzymatic reaction are normally found only in the inner leaflet of the outer membrane, PagP activity may depend on the aberrant migration of phospholipids into the outer leaflet . This finding is consistent with an emerging paradigm for outer membrane enzymes in providing an adaptive response toward disturbances in the outer membrane. Protein Expr Purif, 2002 Oct, 26(1), 59 - 64 Cloning and expression of functional shikimate dehydrogenase (EC 1.1.1.25) from Mycobacterium tuberculosis H37Rv; Magalhaes ML et al.; Tuberculosis (TB), caused by Mycobacterium tuberculosis, continues to be one of the deadliest diseases in the world . TB resurged in the late 1980s and now kills more than 2 million people a year . Possible factors underlying the reemergence of TB are the high susceptibility of human immunodeficiency virus-infected persons to the disease, the proliferation of multi-drug-resistant (MDR) strains, patient noncompliance in completing the standard "short-course" therapy, and decline of health care systems . Thus, there is a need for the development of new antimycobacterial agents to treat MDR strains of M . tuberculosis, to provide for more effective treatment of latent tuberculosis infection, and to shorten the treatment course to improve patient compliance . The shikimate pathway is an attractive target for antimicrobial agents development because it is essential in algae, higher plants, bacteria, and fungi, but absent in mammals . Homologs to enzymes in the shikimate pathway have been identified in the genome sequence of M . tuberculosis . The M . tuberculosis aroE-encoded shikimate dehydrogenase was PCR amplified, cloned, sequenced, and expressed in Escherichia coli BL21(DE3) . Recombinant protein expression was achieved by a low-cost and simple protocol . Although cell lysis resulted in the formation of insoluble aggregates of the recombinant protein, soluble and functional M . tuberculosis shikimate dehydrogenase could be obtained by repeated cycles of freezing and thawing . Enzyme activity measurements demonstrated that there was approximately a 5-fold increase in specific activity for M . tuberculosis shikimate dehydrogenase . Moreover, the enzyme activity was linearly dependent upon the amount of recombinant protein added to the assay mixture, thus, confirming cloning and expression of functional mycobacterial shikimate. Biopharm Drug Dispos, 2002 Oct, 23(7), 293 - 300 Pharmacokinetics and tissue distribution of intravenous pefloxacin for antibiotic prophylaxis in biliary surgery; Gascon AR et al.; The plasma levels and tissue penetration of pefloxacin were studied after prophylactic administration to patients undergoing elective biliary surgery . Pefloxacin was administered as a single dose of 800 mg given intravenously as an infusion 1 h before surgery . Over a period of two years, cultures of bile and stone were performed after cholecystectomy in order to find the main pathogens present in the geographical area of the hospital of Txagorritxu (Vitoria, Spain), as well as to test the antimicrobial susceptibility of these bacteria to pefloxacin . Thirty seven per cent of the bile and stone cultures were positive, and 75 different species were isolated . E . coli was the predominant microorganism (25%) . Other frequent microorganisms were E . faecium (9.3%), S . epidermidis (6.6%) and Cl . perfringens (6.6%) . Most species isolated were susceptible to pefloxacin, with MIC(90) values of 0.125 microg/ml for E . coli, 0.5 microg/ml for S . epidermidis and 1 microg/ml for Cl . perfringens . E . faecium was resistant, with a MIC(90) value of 8 microg/ml but a MIC(50) of 4 microg/ml (intermediate) . After pefloxacin infusion, adequate drug plasma levels (>MIC(90)) for the most frequent pathogens were found throughout the procedure . Elimination half-life was estimated as 22.03+/-6.91 h; the area under the concentration-time curve from zero to infinite had a value of 275.07+/-130.02 mg h/l and the values for volume of distribution at steady-state and plasma clearance were 96.48+/-28.65 L and 3.60+/-1.83 l/h, respectively . Bile pefloxacin concentrations generally exceeded the minimum inhibitory concentrations for most relevant pathogens . Drug levels in gallbladder and subcutaneous tissues were also above the MIC(90) for extended periods . Patients were observed daily throughout their hospital stay . This included examination of the surgical wound and recording of body temperature . No cases of anaerobic infection were noted in the study patients . Other constants such as hospitalization stay and time of recuperation were normal for this type of surgery . According to these results, pefloxacin presents many features that make it suitable for use as a therapeutic prophylactic agent, such as its broad spectrum of antimicrobial activity and favorable pharmacokinetic properties . Clin Infect Dis, 2002 Oct 15, 35(8), 982 - 9 Epub 2002 Sep 24. Has antifungal susceptibility testing come of age? Rex JH, Pfaller MA. The in vitro susceptibility of an infecting organism to the antimicrobial agent selected for therapy is one of several factors that influence the likelihood that therapy for an infection will be successful . To appreciate the value of antifungal susceptibility testing, it is helpful to review the overall predictive utility of antibacterial susceptibility testing . After >30 years of study, in vitro susceptibility can be said to predict the response of bacterial infections with an accuracy that is well summarized as the "90-60 rule": infections due to susceptible isolates respond to therapy approximately 90% of the time, whereas infections due to resistant isolates respond approximately 60% of the time . On the basis of a growing body of knowledge, standardized susceptibility testing for selected organism-drug combinations (most notably, Candida species and the azole antifungal agents) has been shown to have similar predictive utility . Antifungal susceptibility testing is now increasingly and appropriately used as a routine adjunct to the treatment of fungal infections. Eur Biophys J, 2002 Oct, 31(6), 428 - 37 Epub 2002 Jun 28. Lipid discrimination in phospholipid monolayers by the antimicrobial frog skin peptide PGLa . A synchrotron X-ray grazing incidence and reflectivity study; Konovalov O et al.; We present a first study using synchrotron grazing incidence diffraction and X-ray reflectivity measurements on mixed phospholipid/peptide monolayers at the air/water interface . The thermodynamic properties of the pure and mixed monolayers were characterized using the classical film balance technique . Surface pressure/potential-area isotherms showed that the antimicrobial frog skin peptide PGLa formed a very stable monolayer with two PGLa molecules per kinetic unit and a collapse pressure of ~22 mN/m . X-ray grazing incidence diffraction indicated that the peptide-dimer formation did not lead to self-aggregation with subsequent crystallite formation . However, the scattering length density profiles derived from X-ray reflectivity measurements yield information on the PGLa monolayer that protrudes into the air phase by about 0.8 nm, suggesting that the peptide is aligned parallel to the air/water interface . The monolayers, composed of disaturated phosphatidylcholines or phosphatidylglycerols, were stable up to 60 mN/m and exhibited a first-order transition from a liquid-expanded to a liquid-condensed state around 10 mN/m . Structural details of the phospholipid monolayers in the presence and absence of PGLa were obtained from synchrotron experiments . Thereby, the X-ray data of distearoylphosphatidylcholine/PGLa can be analyzed by being composed of the individual components, while the peptide strongly perturbed the lipid acyl chain order of distearoylphosphatidylglycerol . These results are in agreement that PGLa mixes at a molecular level with negatively charged lipids, but forms separate islands in zwitterionic phosphatidylcholine monolayers and demonstrates that antimicrobial peptides can discriminate between the major phospholipid components of bacterial and mammalian cytoplasmic membranes. J Clin Microbiol, 2002 Oct, 40(10), 3845 - 7 Reduction of contamination of mycobacterial growth indicator tubes with a modified antimicrobial combination; Chang CL et al.; Culture in the fluorimetric Mycobacteria Growth Indicator Tube (MGIT) treated with a combination of vancomycin, amphotericin B, and nalidixic acid (VAN) showed growth of most strains of 31 mycobacterial species with a less-than-1-day delay