Antimicrobial

Blood, 2004 Mar 15, 103(6), 2407 - 9 Epub 2003 Nov 20.
Screening hepcidin for mutations in juvenile hemochromatosis: identification of a new mutation (C70R); Roetto A et al.; Juvenile or type 2 hemochromatosis (JH) is a genetic disease caused by increased intestinal iron absorption that leads to early massive iron overload . The main form of the disease is caused by mutations in a still unknown gene on chromosome 1q . Recently, we recognized a second type of JH with clinical features identical to the 1q-linked form, caused by mutations in the gene encoding hepcidin (HEPC) . Hepcidin is a hepatic antimicrobial-like peptide whose role in iron homeostasis was first defined in animal models; deficiency of hepcidin in mice leads to iron overload, whereas its hepatic overexpression in transgenic animals causes iron deficiency . To define the prevalence of HEPC mutations in JH we screened the HEPC gene for mutation in 21 unrelated JH subjects . We identified a new mutation (C70R), which affects 1 of the 8 conserved cysteines that form the disulfide bonds and are critical for the stability of the polypeptide.

Farmaco, 2003 Dec, 58(12), 1235 - 42
Functionalized alkyl and aryl diselenides as antimicrobial and antiviral agents: synthesis and properties; Wojtowicz H et al.; The different dialkyl and diaryl diselenides with carbamoyl and sulfamoyl moieties 2, 3, 5 and other substituents in the ortho position of benzene ring 4, 7, 8 as well as derivatives of 1,2,4-benzoselenadiazine (6) were designed as antiviral and antimicrobial agents and synthesized . Some of them, particularly 8a and 8b, were found in the antiviral assay in vitro to be strong inhibitors of cytopathic activity encephalomyocarditis virus (EMCV) . The compound 4a and 8a were found to have a broad spectrum of acivity against bacteria, yeasts and pathogenic fungi in vitro.

Fitoterapia, 2003 Dec, 74(7-8), 729 - 31
Antimicrobial activity and constituents of Coccoloba acrostichoides; Cota BB et al.; The ethanol extract and fractions from Coccoloba acrostichoides aerial parts were assayed for in vitro antimicrobial activity . The extract was active against the assayed bacteria while most of the fractions also inhibited fungal growth, especially the n-hexane and EtOAc fractions . The isolated beta-sitosterol and betulin were tested, being the last one active against Fusarium oxysporum.

Fitoterapia, 2003 Dec, 74(7-8), 706 - 9
Antimicrobial and cytotoxicity evaluation of Buchholzia coriacea stem bark; Ajaiyeoba EO et al.; Fractions prepared from the methanol extract of Buchholzia coriacea stem bark exhibited a high concentration-dependent antibacterial and antifungal activity compared to the standard antibiotics, ampicillin and tioconazole . In the brine shrimp lethality (BSL) assay, the methanol extract was found to be non-toxic with an LC(50) of 1031 microg/ml . The two main compounds present in the most active fraction were isolated and identified as lupeol and beta-sitosterol.

Fitoterapia, 2003 Dec, 74(7-8), 702 - 5
Antimicrobial activity of aqueous extracts and of berberine isolated from Berberis heterophylla; Freile ML et al.; The antimicrobial activity of Berberis heterophylla leaves, stems and root aqueous extracts was studied in vitro on Gram-positive and Gram-negative bacteria and fungi . The in vitro antifungal activity of berberine isolated from the same source against different Candida species was also investigated.

Fitoterapia, 2003 Dec, 74(7-8), 695 - 8
Antimicrobial activity of extractives of Sarcocephalus coadunatus; Khan MR et al.; The methanolic extracts and the fractions (petrol, dichloromethane, ethyl acetate, butanol) obtained from the leaves, seeds, stem and root barks of Sarcocephalus coadunatus exhibited a high level of broad spectrum antibacterial activity . The activity was more pronounced in the dichloromethane, ethyl acetate and butanol fractions of the leaves; ethyl acetate and butanol fractions of the seeds; dichloromethane fractions of the stem bark and the ethyl acetate fractions of the root bark . None of the fractions showed any antifungal activity.

Fitoterapia, 2003 Dec, 74(7-8), 692 - 4
Antimicrobial activity of Andrographis paniculata; Singha PK et al.; The antimicrobial activity of aqueous extract, andrographolides and arabinogalactan proteins from Andrographis paniculata were evaluated . The aqueous extract showed significant antimicrobial activity, which may be due to the combined effect of the isolated arabinogalactan proteins and andrographolides.

Biotechniques, 2003 Nov, 35(5), 1060 - 4
Antibiotic-free bacterial strain selection using antisense peptide nucleic acid; Dryselius R et al.; Antibiotics are widely useful in medicine, agriculture, and industrial fermentations . However, increasing problems with resistant strains call for restrained use and alternative strategies . Antisense peptide nucleic acids (PNAs) show potent bactericidal effects when targeted against the essential Escherichia coli acpP gene . Aside from attractive antimicrobial therapeutic possibilities for such antisense PNAs, we considered that they could be used as a substitute for antibiotics in bacterial strain selection . Here, treatment of a mixture of E . coli wild-type cells and cells carrying a binding-site altered copy of acpP (acpP-1) with anti-acpP PNA completely killed wild-type cells within 2 h, whereas cells carrying acpP-1 proliferated . Furthermore, electrotransformation of E . coli cells with the plasmid carrying acpP-1 followed by PNA selection gave rise to only true transformants . Unlike previous antibiotic-free selection strategies, this procedure does not require special growth environments or special host strains . Also, the PNA-selected cells grow at a near normal rate . The results open possibilities to use antisense PNAs for strain selection and construction in research and industrial application.

Biotechnol Appl Biochem . 2003 Nov 20; {Epub ahead of print}
Recombinant antimicrobial peptides efficiently produced using novel cloning and purification processes; Metlitskaia L et al.; Endogenous antimicrobial peptides are ubiquitous components of animal and plant host defences . These peptides, usually cationic and amphipathic, kill target cells rapidly and are efficacious against antibiotic-resistant and clinically-relevant pathogens . A practical challenge in the development of cationic peptides as therapeutics is to meet the production requirements for large quantities of highly purified drug substance at competitive costs . While chemical peptide synthesis can be used to manufacture cationic peptides, we have developed cost-effective methods for recombinant production by expressing fusion proteins comprised of multiple copies of the peptides . The fusion proteins accumulate in E . coli inclusion bodies and constitute over 50% of the total cellular proteins . Active antimicrobial peptides are released by chemical reagents and purified by chromatography, combining both standard and novel approaches . Challenges of industrial scale manufacturing of therapeutics were considered in the development of this process.

Int J Hyg Environ Health, 2003 Oct, 206(6), 465 - 72
State-of-the-art hand hygiene in community medicine; Kampf G; Hand hygiene becomes more important in community medicine not only since antibiotic resistant bacteria such as MRSA spread within the community . Hands may be colonized with transient microorganism in up to 75% . Among those transient pathogens S . aureus, C . difficile or the hepatitis C virus may be found . During patient care the number of microorganisms on the hands steadily increases . In addition hands may be contaminated with different kinds of germs even if only "clean" activities are carried out . Gloves may be worn but do not provide complete protection from contamination due to leaks . Therefore hands should always be treated after gloves are taken off . State-of-the-art treatment of hands is the hygienic hand disinfection with alcohol-based hand rubs . They are more effective, quicker to carry out, better tolerated by the skin, with a positive effect on compliance, and cost effective in comparison to antiseptic soaps based on chlorhexidine or triclosan and in comparison to normal non-medicated soaps . Healthy skin easily tolerates alcohol-based products from the beginning on . Only health care workers with an underlying irritative contact dermatitis which is often caused by bar or liquid antiseptic soaps may have difficulties to use alcohol-based products initially . In such a case treatment of the underlying skin condition is the way to go and not staying with a preparation which has caused the dermatitis . All this knowledge is now reflected in current guidelines on hand hygiene . Beside liquids alcohol-based gels can be used if they have an antimicrobial activity equal to alcohol-based liquid preparations . Hand hygiene remains the single most important tool to avoid cross transmission of microorganisms between patients . This state-of-the-art hand hygiene should also be emphasized more in community medicine . This review may help to go the first step into this direction.

Cas Lek Cesk, 2003 Aug, 142(8), 483 - 6
{Treatment of Helicobacter pylori infections in gastric and duodenal ulcers}; Svestka T; Helicobacter pylori is an organism that is thought to be important in the pathophysiology of ulcer disease and gastritis . Eradication of the organism is useful in the treatment of infected patients . Efficacious regimens generally include an antisecretory agent combined with two antimicrobials . The main determinant of the overall cost of treatment is the eradication of H . pylori in the microorganism . Resistance to the commonly used antibiotics can occur but it can be usually overcome with regimen changes . It is important for care physicians to clearly understand when and how to test and how to select appropriate therapy for Helicobacter pylori infection.

Cas Lek Cesk, 2003 Aug, 142(8), 465 - 9
{Hepcidin--a peptide regulating the quantity and distribution of iron in the body in healthy and disease states}; Vokurka M et al.; Iron is an essential element and its amount and balance must be precisely regulated . Iron intestinal absorption is essential for the iron balance; however, the precise mechanism of its regulation remains unknown . Antimicrobial peptide hepcidin, produced in the liver, is considered as a key regulator of iron absorption and kinetics in the organism . Its expression increases in response to the iron overload . Hepcidin decreases iron absorption in the duodenum and causes its sequestration in macrophages . Apart from the iron, inflammation increases hepcidin expression in the liver, and hepcidin is considered to be acute phase protein . Hepcidin is not only the physiological regulator of iron kinetics but is supposed to be a part of the pathogenetic mechanism of anaemia accompanying chronic diseases and its relationship to the hereditary hemochromatosis is also studied.

Cell Mol Life Sci, 2003 Nov, 60(11), 2409 - 26
Pharmacologically active spider peptide toxins; Corzo G et al.; Advances in mass spectrometry and peptide biochemistry coupled to modern methods in electrophysiology have permitted the isolation and identification of numerous novel peptide toxins from animal venoms in recent years . These advances have also opened up the field of spider venom research, previously unexplored due to methodological limitations . Many peptide toxins from spider venoms share structural features, amino acid composition and consensus sequences that allow them to interact with related classes of cellular receptors . They have become increasingly useful agents for the study of voltage-sensitive and ligand-gated ion channels and the discrimination of their cellular subtypes . Spider peptide toxins have also been recognized as useful agents for their antimicrobial properties and the study of pore formation in cell membranes . Spider peptide toxins with nanomolar affinities for their receptors are thus promising pharmacological tools for understanding the physiological role of ion channels and as leads for the development of novel therapeutic agents and strategies for ion channel-related diseases . Their high insecticidal potency can also make them useful probes for the discovery of novel insecticide targets in the insect nervous system or for the development of genetically engineered microbial pesticides.

FEMS Immunol Med Microbiol, 2003 Nov 28, 39(2), 155 - 61
Non-specific immunity-enhancing effects of tryptic casein hydrolysate versus Fermosorb for treatment/prophylaxis of newborn calf colibacillosis; Biziulevicius GA et al.; The effects of treatment/prophylaxis of newborn calf colibacillosis with tryptic casein hydrolysate (TCH), recently shown to be a novel type of antimicrobial acting through stimulation of the microbial autolytic system, versus an authorized veterinary drug, Fermosorb, were evaluated . Both products showed similar high therapeutic and prophylactic efficacies, but hematological indices and daily weight gain of cured/protected animals were better with TCH . The differences in hemoglobin and hematocrit levels, total protein, gamma-globulin and sulfhydryl group quantities, bactericidal and lysozyme activities as well as daily weight gain at the end of treatment/prophylaxis were statistically significant (P<0.05-0.000005) . Statistically significant differences (P<0.05-0.0005) in favor of TCH were also observed when bactericidal activity, total protein quantity of serum as well as daily weight gain of the animals were compared on the 90th day after birth . We conclude that TCH acts not only as an antimicrobial, but also as an immunostimulant (and growth promoter) . The immunostimulatory activity of TCH most probably derives from a synergistic action of bioactive peptides encrypted in the preparation itself and the cell wall fragments resulting from microbial autolysis induction.

Curr Opin Infect Dis, 2003 Dec, 16(6), 547 - 51
Development of drugs for antimicrobial-resistant pathogens; Powers JH; PURPOSE OF REVIEW: Clinicians have noted an association between antimicrobial resistance and antimicrobial use since the introduction of these agents over 50 years ago . The problem of resistance becomes more pressing, however, when organisms acquire resistance mechanisms to multiple antimicrobial agents . Treatment options are limited for some multidrug-resistant organisms . Antimicrobial resistance is a driving force for the need for new antimicrobial agents, especially for these multidrug-resistant pathogens . At the same time, large pharmaceutical companies have indicated that they are devoting fewer resources to antimicrobial drug development . RECENT FINDINGS: This article will review initiatives by the US Food and Drug Administration to identify problem pathogens for which drug development is of most public health importance, and to streamline the drug development process for antimicrobial agents . This article will also touch upon initiatives by federal agencies to implement programs to educate clinicians and the public on the appropriate use of antimicrobial agents to preserve the usefulness of currently marketed drugs . SUMMARY: The most effective way to address the issues of antimicrobial resistance appears to be striking a balance between promoting new drug development and the prudent use of older agents to preserve the usefulness of currently marketed products.

Curr Opin Infect Dis, 2003 Dec, 16(6), 515 - 9
Role of oral antimicrobial therapy in the management of osteomyelitis; Shuford JA et al.; PURPOSE OF REVIEW: Medical therapy of chronic osteomyelitis is largely based on experimental models, historical observational or non-randomized studies, and expert opinions . A minimum of 4-6 weeks of intravenous antimicrobial therapy targeting the causative organism, given in conjunction with surgery, has become the standard for chronic long-bone osteomyelitis in adults . Given the expense, inconvenience, and potential complications inherent to such a treatment program, alternative strategies including effective oral antimicrobial regimens are desirable . RECENT FINDINGS: Several oral antimicrobial agents have undergone evaluation for the treatment of acute and chronic osteomyelitis recently . These include fluoroquinolones, clindamycin, and linezolid . For the treatment of atypical causes of Gram-positive osteomyelitis, other oral therapies have been evaluated with reported success in small numbers of patients . SUMMARY: The standard of care for chronic osteomyelitis in adults remains intravenous antimicrobial therapy, in combination with surgery, for at least 4-6 weeks . Acute osteomyelitis in the pediatric population as well as osteomyelitis caused by atypical Gram-positive organisms and some Gram-negative organisms may be treated successfully with oral antibiotics . Some antimicrobials have equivalent concentration in serum whether administered orally or parenterally . When therapy with these antimicrobials is indicated, the oral route is preferred in compliant patients . As research continues in this area and as new drug formulations are developed, oral therapy may become an accepted alternative in additional selected patients.

Proc Natl Acad Sci U S A, 2003 Nov 25, 100(24), 14281 - 6 Epub 2003 Nov 17.
Delivery of antimicrobials into parasites; Samuel BU et al.; To eliminate apicomplexan parasites, inhibitory compounds must cross host cell, parasitophorous vacuole, and parasite membranes and cyst walls, making delivery challenging . Here, we show that short oligomers of arginine enter Toxoplasma gondii tachyzoites and encysted bradyzoites . Triclosan, which inhibits enoyl-ACP reductase (ENR), conjugated to arginine oligomers enters extracellular tachyzoites, host cells, tachyzoites inside parasitophorous vacuoles within host cells, extracellular bradyzoites, and bradyzoites within cysts . We identify, clone, and sequence T . gondii enr and produce and characterize enzymatically active, recombinant ENR . This enzyme has the requisite amino acids to bind triclosan . Triclosan released after conjugation to octaarginine via a readily hydrolyzable ester linkage inhibits ENR activity, tachyzoites in vitro, and tachyzoites in mice . Delivery of an inhibitor to a microorganism via conjugation to octaarginine provides an approach to transporting antimicrobials and other small molecules to sequestered parasites, a model system to characterize transport across multiple membrane barriers and structures, a widely applicable paradigm for treatment of active and encysted apicomplexan and other infections, and a generic proof of principle for a mechanism of medicine delivery.

Int J Food Microbiol, 2003 Dec 31, 89(2-3), 163 - 70
Comparison of the activity of antifungal hexapeptides and the fungicides thiabendazole and imazalil against postharvest fungal pathogens; Lopez-Garcia B et al.; In this study, we evaluated the activity of short antimicrobial peptides against different fungal isolates that cause postharvest decay of fresh fruits . The previously identified hexapeptides PAF19, PAF26 and LfcinB4-9 inhibited the in vitro growth of isolates from Penicillium digitatum and P . italicum, and from Alternaria and Geotrichum genera, being no active against Rhizopus, Mucor and Aspergillus . The results extend our previous observations on the specific and distinct activity profiles of this class of antifungal peptides . In addition, peptide activities were compared with that of two fungicides used for citrus fruit preservation, thiabendazole (TBZ) and imazalil (IMZ) . We observed a lack of correlation between peptide and fungicide sensitivity among different species . Importantly, P . digitatum and P . italicum isolates resistant to fungicides were susceptible to peptides and our data suggest that common multiple drug resistance mechanisms are not active against this class of peptides . The in vitro peptide inhibition was correlated with a retard of the decay caused by Penicillium on citrus fruits, and this effect was comparable for both fungicide-resistant and -sensitive isolates . Comparison of PAF26 and TBZ in vitro minimum inhibitory concentration (MIC) values and their in vivo effect on citrus decay indicated that PAF26 performed in vivo better than TBZ.

Int J Food Microbiol, 2003 Dec 31, 89(2-3), 125 - 38
Combining nonthermal technologies to control foodborne microorganisms; Ross AI et al.; Novel nonthermal processes, such as high hydrostatic pressure (HHP), pulsed electric fields (PEFs), ionizing radiation and ultrasonication, are able to inactivate microorganisms at ambient or sublethal temperatures . Many of these processes require very high treatment intensities, however, to achieve adequate microbial destruction in low-acid foods . Combining nonthermal processes with conventional preservation methods enhances their antimicrobial effect so that lower process intensities can be used . Combining two or more nonthermal processes can also enhance microbial inactivation and allow the use of lower individual treatment intensities . For conventional preservation treatments, optimal microbial control is achieved through the hurdle concept, with synergistic effects resulting from different components of the microbial cell being targeted simultaneously . The mechanisms of inactivation by nonthermal processes are still unclear; thus, the bases of synergistic combinations remain speculative . This paper reviews literature on the antimicrobial efficiencies of nonthermal processes combined with conventional and novel nonthermal technologies . Where possible, the proposed mechanisms of synergy is mentioned.

Biochem Biophys Res Commun, 2003 Nov 28, 311(4), 853 - 63
AML-1, PU.1, and Sp3 regulate expression of human bactericidal/permeability-increasing protein; Lennartsson A et al.; Bactericidal/permeability-increasing protein (BPI) is an antimicrobial protein in neutrophils, stored in azurophil granules . Expression of BPI is absent in neutrophils of newborns and patients with secondary granule deficiency (SGD), possibly contributing to dysfunction of neutrophils . We report two alternative transcription start sites at 52 and 22bp upstream of the translation start . A proximal 222bp promoter conferring expression in myeloid cells was identified, and critical cis-acting sites for myeloid expression were contained within the 159bp upstream of translation start . Within this region, direct binding and transactivation by AML-1, PU.1, and Sp3 were demonstrated, as judged by electrophoretic mobility shift analysis . Moreover, transient transfections of C/EBPalpha or C/EBPepsilon to HeLa cells resulted in increased promoter activity, indicating a direct or indirect role for C/EBP . In conclusion, we provide evidence for AML-1, PU.1, and Sp3 cooperatively and directly mediating BPI-expression during myeloid differentiation.

J Feline Med Surg, 2003 Dec, 5(6), 305 - 11
Efficacy of azithromycin for the treatment of feline chlamydophilosis; Owen WM et al.; The current recommended treatment for feline chlamydophilosis involves daily oral administration of antimicrobials to all cats within an affected group for a prolonged period of time (4-6 weeks) . Not surprisingly, owner compliance can be poor resulting in apparent treatment failure . Recent anecdotal evidence, supported by its efficacy in the treatment of Chlamydia trachomatis infection in humans, has suggested that azithromycin may offer an alternative by allowing less frequent dosing for a shorter duration . A clinical trial was designed to evaluate the efficacy of azithromycin for the treatment of chlamydia (Chlamydophila felis) infection in cats . Whilst azithromycin, given at 10-15 mg/kg daily for 3 days and then twice weekly, provided a similar, rapid resolution of clinical signs and negative isolation scores as doxycycline, C felis was re-isolated in four out of the five cats treated . Furthermore, even daily administration of azithromycin to chronically infected cats was ineffective in clearing infection . The azithromycin protocols used here were therefore found to be unsuccessful in eliminating the carriage of this strain of C felis.

J Hosp Infect, 2003 Nov, 55 Suppl 1, 1 - 12
Appropriate antimicrobial treatment in nosocomial infections-the clinical challenges; Masterton R et al.; Resistance to antimicrobial agents is emerging in a wide variety of pathogens, particularly those that cause nosocomial infections . As a consequence of this increasing resistance, morbidity and mortality in nosocomial infections is also increasing . It is therefore critical to treat nosocomial infections appropriately by starting antimicrobial treatment early in the course of infection, using the correct agent, at the most appropriate dose, and for an adequate duration . Indeed, early 'appropriate' antibiotic prescribing has been shown significantly to reduce mortality, length of intensive care unit and hospital stay and overall costs.Early use of the correct antibiotic at the appropriate dose and for an adequate duration are key to initial appropriate antibiotic prescribing . Choosing the right antibiotic depends mainly on the likely pathogen(s) and the expected local susceptibility patterns . Selection of appropriate antimicrobial therapy requires a thorough understanding of the likely microbial cause of the infection, including local susceptibility patterns, as well as the properties of the antimicrobials available for treating these infections, namely spectrum of activity and potency (including activity versus known resistance mechanisms), pharmacokinetic profile and tolerability and safety . This review, based on a series of presentations at the 5th International Conference of the Hospital Infection Society (Edinburgh, 2002) examines the importance of appropriate antimicrobial therapy in nosocomial infections, and provides guidance on how to achieve this.

Eur J Biochem, 2003 Nov, 270(22), 4478 - 87
Bilayer localization of membrane-active peptides studied in biomimetic vesicles by visible and fluorescence spectroscopies; Sheynis T et al.; Depth of bilayer penetration and effects on lipid mobility conferred by the membrane-active peptides magainin, melittin, and a hydrophobic helical sequence KKA(LA)7KK (denoted KAL), were investigated by colorimetric and time-resolved fluorescence techniques in biomimetic phospholipid/poly(diacetylene) vesicles . The experiments demonstrated that the extent of bilayer permeation and peptide localization within the membrane was dependent upon the bilayer composition, and that distinct dynamic modifications were induced by each peptide within the head-group environment of the phospholipids . Solvent relaxation, fluorescence correlation spectroscopy and fluorescence quenching analyses, employing probes at different locations within the bilayer, showed that magainin and melittin inserted close to the glycerol residues in bilayers incorporating negatively charged phospholipids, but predominant association at the lipid-water interface occurred in bilayers containing zwitterionic phospholipids . The fluorescence and colorimetric analyses also exposed the different permeation properties and distinct dynamic influence of the peptides: magainin exhibited the most pronounced interfacial attachment onto the vesicles, melittin penetrated more into the bilayers, while the KAL peptide inserted deepest into the hydrophobic core of the lipid assemblies . The solvent relaxation results suggest that decreasing the lipid fluidity might be an important initial factor contributing to the membrane activity of antimicrobial peptides.

Biochemistry, 2003 Nov 25, 42(46), 13725 - 34
Immobilization and aggregation of the antimicrobial peptide protegrin-1 in lipid bilayers investigated by solid-state NMR; Buffy JJ et al.; The dynamics and aggregation of a beta-sheet antimicrobial peptide, protegrin-1 (PG-1), are investigated using solid-state NMR spectroscopy . Chemical shift anisotropies of F12 and V16 carbonyl carbons are uniaxially averaged in 1,2-dilauryl-sn-glycero-3-phosphatidylcholine (DLPC) bilayers but approach rigid-limit values in the thicker 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphatidylcholine (POPC) bilayers . The Calpha-Halpha dipolar coupling of L5 is scaled by a factor of 0.16 in DLPC bilayers but has a near-unity order parameter of 0.96 in POPC bilayers . The larger couplings of PG-1 in POPC bilayers indicate immobilization of the peptide, suggesting that PG-1 forms oligomeric aggregates at the biologically relevant bilayer thickness . Exchange NMR experiments on F12 (13)CO-labeled PG-1 show that the peptide undergoes slow reorientation with a correlation time of 0.7 +/- 0.2 s in POPC bilayers . This long correlation time suggests that in addition to aggregation, geometric constraints in the membrane may also contribute to PG-1 immobilization . The PG-1 aggregates contact both the surface and the hydrophobic center of the POPC bilayer, as determined by (1)H spin-diffusion measurements . Thus, solid-state NMR provides a wide range of information about the molecular details of membrane peptide immobilization and aggregation in lipid bilayers.

Biochemistry, 2003 Nov 25, 42(46), 13613 - 24
SpaC and NisC, the cyclases involved in subtilin and nisin biosynthesis, are zinc proteins; Okeley NM et al.; Lantibiotics are peptide-derived antimicrobial agents that are ribosomally synthesized and posttranslationally modified by a multienzyme complex to their biologically active forms . Nisin has attracted much attention recently due to its novel mechanism of action including specific binding to the bacterial cell wall precursor lipid II, followed by membrane permeabilization . Nisin has been commercially used as a food preservative, while other lantibiotics show promising activity against bacterial infections . The posttranslational modifications are believed to be carried out by a multienzyme complex . At present the enzymes catalyzing the formation of the lantibiotic signature structural motifs, dehydroalanine (Dha), dehydrobutyrine (Dhb), lanthionine (Ln), and methyllanthionine (MeLn), are poorly characterized . In an effort to gain insight into the mechanism by which lantibiotics are biosynthesized, the cyclase enzymes involved in the synthesis of nisin and subtilin (NisC and SpaC, respectively) have been cloned, expressed, and purified . Both proteins exist as monomers in solution and contain a stoichiometric zinc atom . EXAFS data on SpaC and a C349A mutant are in line with two cysteine ligands to the metal in the wild-type enzyme with possibly two additional histidines . The two cysteine ligands are likely Cys303 and Cys349 on the basis of sequence alignments and EXAFS data . The metal may function to activate the cysteine thiol of the peptide substrate toward intramolecular Michael addition to the dehydroalanine and dehydrobutyrine residues in the peptide.

J Am Vet Med Assoc, 2003 Nov 1, 223(9), 1306 - 10, 1280-1
Concurrent bartonellosis and babesiosis in a dog with persistent thrombocytopenia; Tuttle AD et al.; A 12-year-old castrated male West Highland White Terrier was referred because of recurrent episodes of collapsing . The dog was mildly anemic and severely thrombocytopenic and had high serum alanine aminotransferase activity . Infection with Bartonella vinsonii (berkhoffii) was initially diagnosed on the basis of serologic testing . Despite treatment with a series of antimicrobials and prolonged use of immunosuppressive drugs, thrombocytopenia persisted . After 5 months of treatment, Babesia canis organisms were seen during examination of a direct blood smear . The dog was treated with imidocarb dipropionate for babesiosis, after which thrombocytopenia resolved, and administration of immunosuppressive drugs was discontinued . Retrospective review of blood smears failed to identify organisms; however, polymerase chain reaction (PCR) analysis of multiple stored blood samples obtained during the 5-month period of persistent thrombocytopenia identified DNA of B . canis vogeli . Babesiosis may cause persistent, unexplained thrombocytopenia in dogs that are not anemic . A PCR assay can facilitate a diagnosis of babesiosis when organisms are not evident or when serologic testing fails to detect Babesia-specific antibodies.

Dig Liver Dis, 2003 Oct, 35(10), 706 - 10
Use of bovine lactoferrin for Helicobacter pylori eradication; Di Mario F et al.; BACKGROUND: One-week triple therapy is the most frequently recommended treatment for Helicobacter pylori infection . Eradication rate is satisfactory, nevertheless is advisable to look for more effective therapies . AIM: To test the efficacy of a standard triple therapy plus bovine lactoferrin in the eradication of H . pylori infection . PATIENTS AND METHODS: One hundred and fifty consecutive H . pylori positive patients, suffering from dyspeptic symptoms were recruited in a 7-day triple therapy open randomised single centre study with rabeprazole, clarithromycin, tinidazole, bovine lactoferrin (group A) or rabeprazole, clarithromycin, tinidazole (group B), or a 10-day therapy with rabeprazole, clarithromycin, tinidazole (group C) . H . pylori status was assessed 8 weeks after the end of the treatment by means of a 13C-urea breath test or a H . pylori stool antigen-test . RESULTS: Eradication rates (intention to treat/per protocol) were: group A (92.2/95.9%), group B (71.2/72.5%) and group C (70.2/75%) . The efficacy of triple therapy added with lactoferrin was significantly higher than other two regimens (p=0.01, intention to treat analysis; p=0.005, per protocol analysis) . CONCLUSION: These results suggest that lactoferrin tested in the present study was effective in curing H . pylori and could be a new agent to assist the antimicrobials in the eradication of the bacterium.

Pharmacotherapy, 2003 Nov, 23(11), 1497 - 507
Chemical and microbiologic aspects of penems, a distinct class of beta-lactams: focus on faropenem; Hamilton-Miller JM; Many beta-lactam antimicrobials were developed between the 1960s and 1980s, with continuing development driven by the emergence of microbial resistance . Penems form a discrete class of beta-lactams that comprises structural hybrids of penicillins (penams) and cephalosporins (cephems) . The chemistry and microbiology of the representative penems MEN 10700, ritipenem, CGP 31608, sulopenem, BRL 42715, and faropenem are reviewed . Particular emphasis is placed on faropenem, which is in late clinical development.

Pharmacotherapy, 2003 Nov, 23(11), 1486 - 96
Management of severe sepsis: integration of multiple pharmacologic interventions; Micek ST et al.; Severe sepsis is an infection-induced process that often promotes organ dysfunction and death in up to 50% of afflicted patients . Clinical advances that improve patient survival include early goal-directed volume resuscitation, broad-spectrum empiric antimicrobial therapy with deescalation strategies, therapy with drotrecogin alfa (activated), glucocorticoid replacement in patients with adrenal insufficiency, and tight control of blood glucose levels . The challenge for critical care practitioners is to integrate the many pharmacologic and supportive interventions required for optimal care of these patients.

Scand J Infect Dis, 2003, 35(9), 670 - 6
Cathelicidins and innate defense against invasive bacterial infection; Nizet V et al.; Cathelicidins are small cationic peptides that possess broad-spectrum antimicrobial activity . These gene-encoded 'natural antibiotics' are produced by several mammalian species on epithelial surfaces and within the granules of phagocytic cells . Since their discovery over a decade ago, cathelicidins have been speculated to function within the innate immune system, contributing to a first line of host defense against an array of microorganisms . Consequently, cathelicidins have captured the interest of basic investigators in the diverse fields of cell biology, immunology, protein chemistry and microbiology . A burgeoning body of experimental research now appears to confirm and extend the biological significance of these fascinating molecules . This article reviews the latest advances in the knowledge of cathelicidin antimicrobial peptides, with particular emphasis on their role in defense against invasive bacterial infection and associations with human disease conditions.

Scand J Infect Dis, 2003, 35(9), 573 - 6
Macrophage migration inhibitory factor and host innate immune responses to microbes; Calandra T; Among innate immune cells, macrophages play an essential role in the sensing and elimination of invasive microorganisms . Binding of microbial products to pathogen-recognition receptors stimulates macrophages to release cytokines and other effector molecules that orchestrate the host innate and adaptive immune responses . Recently, the protein known as macrophage migration inhibitory factor (MIF) has emerged as a pivotal mediator of innate immunity . First identified as a T-cell cytokine, MIF was rediscovered as a protein released by pituitary cells after exposure to endotoxin {lipopolysaccharide (LPS)} or bacteria and in response to stress . Monocytes, macrophages and lymphocytes constitutively express MIF, which is rapidly released after stimulation with bacterial endotoxins and exotoxins, and cytokines . MIF induces powerful proinflammatory biological responses and has been shown to be an important effector molecule of septic shock . High levels of MIF have been detected in the circulation of patients with severe sepsis and septic shock . Inhibition of MIF activity with neutralizing anti-MIF antibodies or deletion of the Mif gene led to a marked reduction in cytokine production and protected mice from lethal bacterial sepsis and toxic shock induced by Gram-negative endotoxin or Gram-positive exotoxins . Investigations into the mechanisms whereby MIF modulates innate immune responses to endotoxin and Gram-negative bacteria have shown that MIF up-regulates the expression of Toll-like receptor 4 (TLR4), the signal-transducing molecule of the LPS receptor complex . Thus, MIF enables cells, such as the macrophage, that are at the forefront of the host antimicrobial defences, to sense promptly the presence of invading Gram-negative bacteria and mount an innate immune response . Given that it is a pivotal regulator of innate immune responses to bacterial infections, MIF appears to be a perfect target for novel therapeutic interventions in patients with severe sepsis.

Am J Health Syst Pharm, 2003 Nov 1, 60(21), 2229 - 32
Neutropenia in patients receiving long-term cefepime therapy for osteomyelitis; Wong BB et al.; The frequency of neutropenia in patients receiving long-term cefepime therapy for osteomyelitis compared with that in patients receiving other antimicrobials was studied . A comparative case review was conducted of home infusion patients receiving cefepime and patients receiving other antimicrobials for osteomyelitis . All courses of antimicrobial therapy for osteomyelitis between January 2001 and December 2002 were evaluated . The duration of prescribed therapy was approximately six weeks . Weekly laboratory tests included complete blood counts with differential . A total of 134 courses of antimicrobial therapy were reviewed (13 courses of cefepime therapy in 12 patients and 121 courses of therapy with other i.v . antimicrobials in 120 patients) . Eight (62%) of the cefepime courses resulted in neutropenia, compared with none in the courses of other antimicrobials . Neutropenia was detected after 17-30 days of cefepime therapy . Blood counts returned to normal within one week of cefepime discontinuation . Eight of 13 courses of extended therapy with i.v . cefepime for osteomyelitis resulted in neutropenia, compared with none of 121 courses of other antimicrobials . Clinicians should exercise extreme caution when prescribing cefepime for longer than 14 days.

Helicobacter, 2003, 8 Suppl 1, 53 - 60
Treatment of Helicobacter pylori infection; Perri F et al.; Review of the recently published data on Helicobacter pylori management highlights various interesting aspects . Current H . pylori eradication guidelines generally suggest a noninvasive 'test and treat' strategy for all dyspeptic patients with certain age limits depending on the local gastric neoplasia risk . According to the 'Maastricht 2-2000 Consensus Report' treatment should be thought of as a 'package' considering first- and second-line eradication therapies together . Various centres have published their results using novel antimicrobial formulations and 'rescue' and 'sequential' therapies . Review suggests that care at the specialist level remains a challenge and guidelines are deficient particularly as regards the selection and duration of eradication therapies . Results indicate that differences for CYP2C19 genotype and the selection of proton pump inhibitors have no significant role in determining eradication rates whereas antibiotic resistance and socio-economic factors play a variable role according to different geographical areas . Compliance remains an important factor in determining clinical outcome at the primary and secondary levels worldwide.

Curr Opin Ophthalmol, 2003 Dec, 14(6), 413 - 9
Retinal vasculitis; Walton RC et al.; Retinal vasculitis represents a group of diseases characterized by inflammation affecting the retinal vasculature . It is an uncommon disorder that may occur as an isolated disease or more commonly in association with other ocular diseases or a variety of systemic diseases . With a wide variety of disease associations, a search for an underlying etiology should be undertaken based on a meticulous history, review of systems, and physical examination . The laboratory evaluation of patients with retinal vasculitis is an essential component of the work-up to facilitate detection of any underlying disease or to establish a limited differential diagnosis . The management of infectious causes of retinal vasculitis consists of antimicrobial therapy while noninfectious retinal vasculitis is managed with corticosteroids and/or immunosuppressive agents . Because retinal vasculitis is an uncommon disease, there are only a limited number of publications over the past year related to this topic.

Pediatr Infect Dis J, 2003 Nov, 22(11), 996 - 1002
Role of antimicrobial applications to the umbilical cord in neonates to prevent bacterial colonization and infection: a review of the evidence; Mullany LC et al.; In developing countries umbilical cord infections constitute a major cause of neonatal morbidity and pose significant risk for mortality, whereas outbreaks of cord infections continue to occur in developed country nurseries . Cord infections in developing countries can be prevented through increasing access to tetanus toxoid immunization during pregnancy, promoting clean cord care and reducing harmful cord applications and behaviors . Interventions introduced in both developed and developing countries to reduce exposure of the cord to infectious pathogens include clean cord cutting, hand-washing before and after handling the baby, bathing of the infant with antimicrobial agents and application of antimicrobials to the cord . Despite the importance of umbilical cord care, both traditionally and medically, there have been few randomized trials investigating the impact of different cord care regimens on rates of local or systemic infections, particularly in developing countries.This review examines available data on umbilical cord care, with a particular focus on those comparing rates of bacterial colonization and/or rates of cord infection among neonates receiving different umbilical cord care regimens . Although most investigators agree that topical antimicrobials reduce bacterial colonization of the cord, a firm relationship between colonization and infection has not been established . Further research in developed countries, including follow-up beyond hospital discharge, is required before advising on "best cord care practices." The paucity of published reports from developing countries indicates the need to investigate the impact of antimicrobial applications on cord and systemic infections in a community-based, prospective manner.

Microbes Infect, 2003 Nov, 5(14), 1317 - 27
Neutrophil granules and secretory vesicles in inflammation; Faurschou M et al.; The neutrophil is a major effector cell of innate immunity . Exocytosis of granules and secretory vesicles plays a pivotal role in most neutrophil functions from early activation to the destruction of phagocytosed microorganisms . Neutrophil granules contain a multitude of antimicrobial and potentially cytotoxic substances that are delivered to the phagosome or to the exterior of the cell following degranulation . This review summarises current knowledge of granule biology and highlights the effects of neutrophil degranulation in the acute inflammatory response.

Microbes Infect, 2003 Nov, 5(14), 1293 - 8
Neutrophil cell signaling in infection: role of phosphatidylinositide 3-kinase; Moraes TJ et al.; Neutrophils play a pivotal role in the innate immune response to microbial pathogens . They are uniquely suited to this role by virtue of specialized antimicrobial capabilities that include the capacity to sense minute amounts of microbial products and inflammatory mediators, to move to the site of infection, and finally to bind, internalize and kill the pathogens . To optimize host defense capabilities while minimizing damage to host tissues ('collateral damage'), these microbicidal responses must be tightly regulated . Additionally, neutrophils clear inflammatory debris, a process that is necessary for restoration of the native architecture and function of the tissue . This review highlights some recent advances in our knowledge of cell signaling as it pertains to neutrophil function, with specific emphasis on the role of the phosphatidylinositide 3-kinase in antimicrobial function.

Bioorg Chem, 2003 Dec, 31(6), 425 - 36
Cathelicidin family of antimicrobial peptides: proteolytic processing and protease resistance; Shinnar AE et al.; Cathelicidins are a gene family of antimicrobial peptides produced as inactive precursors . Signal peptidase removes the N-terminal signal sequence, while peptidylglycine alpha-amidating monooxygenase often amidates and cleaves the C-terminal region . Removal of the cathelin domain liberates the active antimicrobial peptide . For mammalian sequences, this cleavage usually occurs through the action of elastase, but other tissue-specific processing enzymes may also operate . Once released, these bioactive peptides are susceptible to proteolytic degradation . We propose that some mature cathelicidins are naturally resistant to proteases due to their unusual primary structures . Among mammalian cathelicidins, proline-rich sequences should resist attack by serine proteases because proline prevents cleavage of the scissile bond . In hagfish cathelicidins, the unusual amino acid bromotryptophan may make the active peptides less susceptible to proteolysis for steric reasons . Such protease resistance could extend the pharmacokinetic lifetimes of cathelicidins in vivo, sustaining antimicrobial activity.

Chembiochem, 2003 Nov 7, 4(11), 1151 - 63
4-fluorophenylglycine as a label for 19F NMR structure analysis of membrane-associated peptides; Afonin S et al.; The non-natural amino acid 4-fluorophenylglycine (4F-Phg) was incorporated into several representative membrane-associated peptides for dual purpose . The (19)F-substituted ring is directly attached to the peptide backbone, so it not only provides a well-defined label for highly sensitive (19)F NMR studies but, in addition, the D and L enantiomers of the stiff side chain may serve as reporter groups on the transient peptide conformation during the biological function . Besides peptide synthesis, which is accompanied by racemisation of 4F-Phg, we also describe separation of the epimers by HPLC and removal of trifluoroacetic acid . As a first example, 18 different analogues of the fusogenic peptide "B18" were prepared and tested for induction of vesicle fusion; the results confirmed that hydrophobic sites tolerated 4F-Phg labelling . Similar fusion activities within each pair of epimers suggest that the peptide is less structured in the fusogenic transition state than in the helical ground state . In a second example, five doubly labelled analogues of the antimicrobial peptide gramicidin S were compared by using bacterial growth inhibition assays . This cyclic beta-sheet peptide could accommodate both L and D substituents on its hydrophobic face . As a third example, we tested six analogues of the antimicrobial peptide PGLa . The presence of d-4F-Phg reduced the biological activity of the peptide by interfering with its amphiphilic alpha-helical fold . Finally, to illustrate the numerous uses of l-4F-Phg in (19)F NMR spectroscopy, we characterised the interaction of labelled PGLa with uncharged and negatively charged membranes . Observing the signal of the free peptide in an aqueous suspension of unilamellar vesicles, we found a linear saturation behaviour that was dominated by electrostatic attraction of the cationic PGLa . Once the peptide is bound to the membrane, however, solid-state (19)F NMR spectroscopy of macroscopically oriented samples revealed that the charge density has virtually no further influence on the structure, alignment or mobility of the peptide.

FEMS Microbiol Lett, 2003 Nov 7, 228(1), 27 - 31
Isolation of a novel antimicrobial peptide gene (Sp-AMP) homologue from Pinus sylvestris (Scots pine) following infection with the root rot fungus Heterobasidion annosum; Asiegbu FO et al.; A new family of antimicrobial peptide homologues termed Sp-Amp has been discovered in Pinus sylvestris (Scots pine) . This is the first report of such proteins to be characterized in a conifer species . Sp-AMP1 was identified in a substructured cDNA library of root tissue infected with the root rot fungus Heterobasidion annosum and encodes a mature peptide of 79 amino acid residues . Three additional members of the Sp-AMP family (Sp-AMPs 2-4) encode cysteine-rich proteins of 105 amino acids, each containing an N-terminal region with a probable cleavage signal sequence . Northern analysis confirmed that Sp-AMP expression is elevated in Scots pine roots upon infection with H . annosum . These peptides share 64% amino acid identity with a mature protein from Macadamia integrifolia (MiAMP1), which allowed us to build a homology model for preliminary analysis . Southern analyses further confirmed that several copies of the gene are present in the Scots pine genome . The potential significance of Sp-AMP in the H . annosum-conifer pathosystem is discussed.

Bioorg Med Chem Lett, 2003 Sep 1, 13(17), 2933 - 6
Synthesis and evaluation of methyl ether derivatives of the vancomycin, teicoplanin, and ristocetin aglycon methyl esters; McComas CC et al.; A series of methyl ether derivatives of the vancomycin, teicoplanin, and ristocetin aglycon methyl esters was synthesized and their antimicrobial activity was established . These derivatives exhibit increased activity against VanB resistant strains of bacteria equipotent with that observed with sensitive bacteria.

Biosens Bioelectron, 2003 Nov 30, 19(3), 269 - 76
The influence of antimicrobial treatments on the cytocompatibility of polyurethane biosensor membranes; von Woedtke T et al.; The cytocompatibility of polyurethane membranes was tested following ultraviolet or gamma irradiation as well as treatment with hydrogen peroxide or glutaraldehyde containing solutions . Despite the fact that all of the methods had been recommended for antimicrobial treatment of glucose biosensors, the treatments investigated significantly influenced cytocompatibility characteristics . Cytotoxicity of membrane eluates was not observed following irradiation treatments . This was also the case when the membranes were repeatedly washed following chemical treatment . Cell growth upon the membranes was stimulated to a different extent after gamma and UV irradiation as well as following hydrogen peroxide treatments . Residues of an urea-based hydrogen peroxide inclusion compound caused a restriction in cell growth upon the membranes as was similarly observed with 2 and 4% glutaraldehyde solutions acting over 2 and 4 h, respectively . It is concluded that cytocompatibility in vitro reflecting the host response against a biomaterial in vivo does not only depend upon the material itself but also upon antimicrobial treatments which could have consequences for its bioperformance characteristics.

Int J Immunopathol Pharmacol, 2003 Sep-Dec, 16(3), 241 - 6
Novel peptides enhance the production of nitric oxide and inducible nitric oxide synthase (iNOS) gene expression in murine macrophage; Acharya A et al.; Bioactive novel polypeptide of anuran skin has a wide range of antimicrobial properties against the infection and tumour cell . Macrophages are known to produce the Nitric oxide (NO) by a variety of cells upon activation . NO produced by the activated macrophages an important mediator for antimicrobial and tumoricidal activity . In-vitro macrophage exposed with medium alone, containing LPS, containing polypepeptides and LPS plus polypeptides for 24 h showed enhanced production of NO with respect to control and LPS treated and significant increase in NO production in LPS plus polypeptide . Western blot and PCR analysis also showed that increased production of protein expression and mRNA expression of inducible nitric oxide synthase (iNOS) . These findings suggest that novel polypeptides are potent activating agent for enhanced production of NO through activation of iNOS gene.

J Burn Care Rehabil, 2003 Nov-Dec, 24(6), 395 - 9
Role of thymus oil in burn wound healing; Dursun N et al.; Thymus oil and its components are becoming increasingly popular as naturally occurring antimicrobial and antioxidant agents . The real importance of thymus on nitric oxide (NO) is unknown . NO is an important mediator in numerous physiologic and pathophysiologic events . Stasis and thrombosis in burn wound can progress as a result of the release of local mediators . The implication of NO in burn injury is not well studied . In this study, we tried to determine the role of burn-induced NO and whether thymus oil plays a protective role after a thermal injury . Rats were divided into five groups . We topically applied thymus oil, olive oil, and silverdin and sulfadiazine on the rats, respectively, during a period of 21 days after they were burned while under anesthesia . The burned control group and nonburned control group did not receive any treatment . The results of this study show that NO was overproduced by thermal injury and decreased during the days after burn injury . The decrease in rats treated with thymus and sulfadiazine was higher than the others . These data indicate that thymus oil may serve as a protective agent to the damaged tissues by decreasing the NO level . Histologic examination results show that the formation of new tissue in rats receiving thymus oil was more than other burned groups, and this finding supports our hypothesis.

Arch Pharm Res, 2003 Oct, 26(10), 773 - 7
Synthesis and antimicrobial activities of some new nitroimidazole derivatives; Benkli K et al.; In this study, some new nitroimidazole derivatives were obtained from 2-(2-methyl-5-nitro-1H-imidazol-1-yl)ethylamine dihydrochloride (4) and 1-(2-bromoethyl)-2-methyl-5-nitroimidazole (5), which were prepared using metronidazole . Compound 4 was reacted with arylisothiocyanates (6) to obtain 1-{2-(2-methyl-5-nitroimidazol-1-yl)ethyl}-3-arylthioureas (7) and the latter with alpha-bromoacetophenones (8) to give 3-{2-(2-methyl-5-nitroimidazol-1-yl)ethyl}-2-arylimino-4-aryl-4-thiazolines (9) . Also 1-{2-(2-methyl-5-nitroimidazol-1-yl)ethyl}-2-phenyl-4-arylideneimidazolin-5-ones (11) were prepared by reaction of 4 with 2-phenyl-4-arylidene-5-oxazolones (10) . The reaction of the other starting material 5 with 5-arylidenethiazolidin-2,4-dione (12) gave 3-{2-(2-methyl-5-nitroimidazol-1-yl)ethyl}-5-arylidenethiazolidin-2,4-dione (13) derivatives . Structural elucidation of the compounds was performed by IR, 1H-NMR and MASS spectroscopic data and elemental analysis results . Antimicrobial activities of the compounds were examined and moderate activity was obtained.

Vet Clin North Am Food Anim Pract, 2003 Nov, 19(3), 625 - 46
Susceptibility testing for bovine respiratory and enteric disease; Apley MD; The interpretation of susceptibility results for antimicrobials with NCCLS-approved veterinary-specific breakpoints and where the methods also were NCCLS-approved are well established . When these same breakpoints are applied to other applications, however, the interpretation is not so clear . In these cases, a finding of S based on serial-dilution breakpoints puts the isolate in a defined population of bacteria with an MIC equal to or below the S breakpoint . An R result, in these cases, indicates that the organism may have an MIC equal to or greater (with no limits) than the R breakpoint . Extended-dilution testing yields more specific information about the isolate MIC . The relationship of disk-diffusion zone diameters to serial-dilution MICs is correlated on the basis of specific bacterial populations . When disk-diffusion results are interpreted for isolates other than those used for interpretive criteria development, the clinician is left wondering if the zone-diameter results now have a different relationship to serial-dilution results . Furthermore, the question of predictive value of the serial-dilution break-points still remains . The veterinary clinician should be aware of the differences in susceptibility testing predictive value for different applications . When approved veterinary-specific interpretive criteria are not available, then it is appropriate to keep records of clinical response related to susceptibility testing results for common therapies . Advice should be sought on the relationship of pathogen MICs to pharmacokinetic-pharmacodynamic parameters in these situations.

Minerva Pediatr, 2003 Oct, 55(5), 415 - 38
Acute otitis media disease management; Pichichero ME et al.; A first step in management decisions regarding otitis media must focus on accurate diagnosis to distinguish normal from acute otitis media (AOM) from otitis media with effusion (OME) or a retracted tympanic membrane without middle ear effusion . There are several classification schemes for AOM that may impact management decisions: patients with acute, persistent, recurrent, or chronic AOM may have a different distribution of bacterial pathogens and a different likelihood of success from antimicrobial therapy . Patient age, prior treatment history and daycare attendance are other important variables . The natural history of AOM without antibiotic treatment is generally favorable; however, from the few studies available, this is difficult to quantitate because the diagnosis was infrequently confirmed by tympanocentesis leaving the possibility that many patients entered into these trials may not have had bacterial AOM . Antibiotic choices should reflect pharmacokinetic/pharmacodynamic data and clinical trial results demonstrating effectiveness in eradication of the most likely pathogens based on tympanocentesis sampling and antibiotic sensitivity testing . Thereafter, compliance factors such as formulation, dosing schedule and duration of treatment and accessibility factors such as availability and cost should be taken into account . The increasing prevalence of antibiotic resistance among AOM pathogens and the changing susceptibility profiles of these bacteria should be considered in antibiotic selection . Current best practice recommends amoxicillin for uncomplicated AOM; continuing or switching to an alternative antibiotic based on clinical response after 48 hours of therapy; and selection of second line antibiotics as first line choices when the patient has already been on an antibiotic within the previous month or is otitis prone . Preferred second-line agents frequently noted in various guidelines include amoxicillin/clavulanate, cefdinir, cefpodoxime, cefprozil, and cefuroxime . Three injections of ceftriaxone or gatifloxacin (when approved) or diagnostic/therapeutic tympanocentisis (when approved) become a third-line treatment option . No single antibiotic or management strategy is ideal for all patients.

J Immunol, 2003 Nov 15, 171(10), 5233 - 43
Timing of IFN-beta exposure during human dendritic cell maturation and naive Th cell stimulation has contrasting effects on Th1 subset generation: a role for IFN-beta-mediated regulation of IL-12 family cytokines and IL-18 in naive Th cell differentiation; Nagai T et al.; Type I IFNs, IFN-alpha and IFN-beta, are early effectors of innate immune responses against microbes that can also regulate subsequent adaptive immunity by promoting antimicrobial Th1-type responses . In contrast, the ability of IFN-beta to inhibit autoimmune Th1 responses is thought to account for some of the beneficial effects of IFN-beta therapy in the treatment of relapsing remitting multiple sclerosis . To understand the basis of the paradoxical effects of IFN-beta on the expression of Th1-type immune responses, we developed an in vitro model of monocyte-derived dendritic cell (DC)-dependent, human naive Th cell differentiation, in which one can observe both positive and negative effects of IFN-beta on the generation of Th1 cells . In this model we found that the timing of IFN-beta exposure determines whether IFN-beta will have a positive or a negative effect on naive Th cell differentiation into Th1 cells . Specifically, the presence of IFN-beta during TNF-alpha-induced DC maturation strongly augments the capacity of DC to promote the generation of IFN-gamma-secreting Th1 cells . In contrast, exposure to IFN-beta during mature DC-mediated primary stimulation of naive Th cells has the opposite effect, in that it inhibits Th1 cell polarization and promotes the generation of an IL-10-secreting T cell subset . Studies with blocking mAbs and recombinant cytokines indicate that the mechanism by which IFN-beta mediates these contrasting effects on Th1 cell generation is at least in part by differentially regulating DC expression of IL-12 family cytokines (IL-12 and/or IL-23, and IL-27) and IL-18.

Biomacromolecules, 2003 Nov-Dec, 4(6), 1811 - 7
Structure-activity relationships of oligoguanidines-influence of counterion, diamine, and average molecular weight on biocidal activities; Albert M et al.; A series of different oligomeric guanidines was prepared by polycondensation of guanidinium salts and four different diamines under varying conditions . The antimicrobial activities were evaluated against two to four microorganisms . MALDI-TOF-MS was used to analyze the different oligomers . It was found that in each case three major product type series are dominating . These series are linear and terminated with one guanidine and one amino group (type A), two amino groups (type B), or two guanidine groups (type C), respectively . By using 1,2-bis(2-aminoethoxy)ethane as the amino component, a considerable amount of two additional product series, consisting of cyclic structures, was detected (type D and E) . It turned out that an average molecular mass of about 800 Da is necessary for an efficient antimicrobial activity . Lower Mw's result in a rapid decrease of activity . By using guanidinium carbonate as the starting material, oligomers with low biocidal activity were obtained, which was caused by incorporation of urea groups during the polycondensation . The diamine determines the distance between two guanidinium groups . It was shown that both 1,2-bis(2-aminoethoxy)ethane and hexamethylenediamine give oligomers with high biocidal activity . By increasing the chain length of the diamine, the biocidal activity drops again.

Biomacromolecules, 2003 Nov-Dec, 4(6), 1457 - 65
Chitosan as antimicrobial agent: applications and mode of action; Rabea EI et al.; Chitosan, a hydrophilic biopolymer industrially obtained by N-deacetylation of chitin, can be applied as an antimicrobial agent . The current review of 129 references describes the biological activity of several chitosan derivatives and the modes of action that have been postulated in the literature . It highlights the applications of chitosan as an antimicrobial agent against fungi, bacteria, and viruses and as an elicitor of plant defense mechanisms.

Drug Dev Ind Pharm, 2003 Oct, 29(9), 1027 - 33
Degradation kinetics of somatostatin in aqueous solution; Herrmann J et al.; The degradation kinetics of somatostatin (somatotropin release inhibiting factor), a cyclic tetradecapeptide, was investigated as a function of temperature, pH, ionic strength, buffer type, and buffer concentration . In addition, the effect of different container materials in which the solutions were stored and the presence of an antimicrobial agent for in vitro use was examined . The degradation of somatostatin followed first-order kinetics under all investigated conditions . The pH-stability profile showed a well-defined stability optimum around pH 3.7 . The degradation was accelerated at higher buffer concentrations, phosphate buffer being significantly more detrimental than acetate buffer . The ionic strength and the drug concentration had virtually no effect on the degradation rate . When general purpose glass vials were used as storage containers, degradation was faster due to release of alkali from the container material . The solution properties, i.e., pH, buffer type, buffer capacity, and the experimental setup such as container material and sterile conditions need to be carefully selected or maintained, in order to avoid accelerated degradation.

J Clin Microbiol, 2003 Nov, 41(11), 5121 - 6
Detection and differentiation of Mycobacterium tuberculosis and nontuberculous mycobacterial isolates by real-time PCR; Shrestha NK et al.; Mycobacteria cause a variety of illnesses that differ in severity and public health implications . The differentiation of Mycobacterium tuberculosis from nontuberculous mycobacteria (NTM) is of primary importance for infection control and choice of antimicrobial therapy . Despite advances in molecular diagnostics, the ability to rapidly diagnose M . tuberculosis infections by PCR is still inadequate, largely because of the possibility of false-negative reactions . We designed and validated a real-time PCR for mycobacteria by using the LightCycler system with 18 reference strains and 168 clinical mycobacterial isolates . All clinically significant mycobacteria were detected; the mean melting temperatures (with 99.9% confidence intervals {99.9% CI} in parentheses) for the different mycobacteria were as follows: M . tuberculosis, 64.35 degrees C (63.27 to 65.42 degrees C); M . kansasii, 59.20 degrees C (58.07 to 60.33 degrees C); M . avium, 57.82 degrees C (57.05 to 58.60 degrees C); M . intracellulare, 54.46 degrees C (53.69 to 55.23 degrees C); M . marinum, 58.91 degrees C (58.28 to 59.55 degrees C); rapidly growing mycobacteria, 53.09 degrees C (50.97 to 55.20 degrees C) or 43.19 degrees C (42.19 to 44.49 degrees C) . This real-time PCR assay with melting curve analysis consistently accurately detected and differentiated M . tuberculosis from NTM . Detection of an NTM helps ensure that the negative result for M . tuberculosis is a true negative . The specific melting temperature also provides a suggestion of the identity of the NTM present, when the most commonly encountered mycobacterial species are considered . In a parallel comparison, both the LightCycler assay and the COBAS Amplicor M . tuberculosis assay correctly categorized 48 of 50 specimens that were proven by culture to contain M . tuberculosis, and the LightCycler assay correctly characterized 3 of 3 specimens that contained NTM.

Bioorg Med Chem, 2003 Nov 17, 11(23), 5035 - 43
Analysis of structural features of bis-quaternary ammonium antimicrobial agents 4,4'-(alpha,omega-polymethylenedithio)bis (1-alkylpyridinium iodide)s using computational simulation; Ohkura K et al.; The bis-quaternary ammonium compounds (QACs) consisted of two identical alkylpyridinium rings and a bridge structure linking the rings to each other . The QACs have a methylene bridge except for 4DCABP-P,12 which has a phenyl ring as a bridge . These bis-QACs are as follows; amide type: N,N'-tetramethylenebis(1-dodecyl-4-carbamoylpyridinium iodide) (4BCAP-4,12), N,N'-hexamethylenebis(1-decyl-4-carbamoylpyridinium iodide) (4BCAP-6,10), anti-amide type: 4,4'-(1,4-tetramethylenedicarbonyldiamine)bis(1-decylpyridinium iodide) (4DCABP-4,10), 4,4'-(1,4-tetramethylenedicarbonyldiamine)bis (1-dodecylpyridinium iodide) (4DCABP-4,12), 4,4'-(1,4-phenyldicarbonyldiamine)bis(1-dodecylpyridinium iodide) (4DCABP-P,12), ester type: 4,4'-(1,6-hexamethylenedioxydicarbonyl)bis(1-dodecylpyridinium iodide) (4DOCBP-6,12), thioether type: 4,4'-(1,6-hexamethylenedithio)bis(1-octylpyridinium iodide) (4DTBP-6,8) . From the investigation of the relationship between the median lethal dose (LD(50)) and the minimum inhibitory concentration (MIC) of these compounds, 4DTBP-6,8 as a disinfectant, seems to be very safe for human cells . The global minimum of 4DTBP-6,8 were searched and 1125 conformers obtained . The solvation free energy (dGW) of nine samples, which were extracted from these 1125 conformers, was calculated and two minimum points of dGW were observed . In the conformer-energy analysis of four types of model bridge-molecule, the thioether type bridge indicated a gradual energy increment, while the other three (amide, anti-amide, ester) types indicated an energy jump point in their profiles . Then we considered that the delicate balance between hydrophobicity and structural feature in the bridge-region of 4DTBP-6,8 molecule seemed to be related to its safety antibacterial activity.

Perit Dial Int, 2003 Sep-Oct, 23(5), 450 - 5
Increased severity of Escherichia coli peritonitis in peritoneal dialysis patients independent of changes in in vitro antimicrobial susceptibility testing; Valdes-Sotomayor J et al.; OBJECTIVE: Despite improvements in peritoneal dialysis (PD) technique, peritonitis continues to be one of the most frequent complications of PD . Nonresolving peritonitis remains a risk for severe anatomical peritoneal changes that may limit the viability of the membrane for dialysis purposes . We have observed remarkably poor outcome of peritonitis caused by Escherichia coli in the past 6 years . With its very low response rate to broad-spectrum antibiotics, the increased severity of E . coli peritonitis deteriorates peritoneal function and affects patient outcome . DESIGN: Retrospective study . SETTING: Two large PD units in two university hospitals . PATIENTS AND METHODS: The total number of patients reviewed was 456 . The records of 49 E . coli peritonitis episodes were studied.The observation period started in 1980 and ended in March 2001 . Sixteen males and 19 females were included . Severity was defined in terms of days of peritoneal inflammation, lack of response to a potentially useful antibiotic, requirement for catheter removal, and/or laparotomy . Study cases (study group) were those episodes appearing after 1996 (when the first severe cases appeared) and historic controls were episodes occurring before 1996 . RESULTS: In the study group, 18 peritonitis episodes developed in 15 patients . In the control group, 31 peritonitis episodes developed in 20 patients . There were no significant differences in clinical presentation; however, the outcome was significantly poorer for the later period . A severe outcome occurred in 50% of study versus 10% of control patients . In fact, 68% of the episodes registered before 1996 were cured in 3 days or less . Concurring with this trend, the numbers of surgical interventions and catheter removals were also higher in the study group . Strikingly, E . coli did not show changes in in vitro susceptibility testing to antibiotics, although the in vivo response was much worse . CONCLUSIONS: We describe a change in the virulence of E . coli peritonitis episodes over the past 5 years leading to a high percentage of treatment failure, which does not depend on antibiotic sensitivity and seems to be dependent on changes in host response mechanisms.

J Clin Pediatr Dent, 2003 Fall, 28(1), 47 - 52
Biological factors in dental caries: role of saliva and dental plaque in the dynamic process of demineralization and remineralization (part 1); Hicks J et al.; Dental caries is a complex disease process that afflicts a large proportion of the world's population, regardless of gender, age and ethnicity, although it does tend to affect more indivduals with a low socioeconomic status to a greater extent . The process of dental caries is dependent upon biological factors that are present within the saliva and dental plaque . There are many different agents within saliva and plaque that serve to protect the tooth surface against caries development . Salivary flow rate, buffering capacity, antimicrobial activity, microorganism aggregation and clearance from the oral cavity, immune surveillance, and calcium phosphate binding proteins all interact to inhibit or reverse demineralization of exposed tooth surfaces . Cariogenic bacteria levels within the saliva and plaque determine whether caries will occur or not, and the concentration in saliva and plaque are intimately related to the type of carbohydrate ingestion and the frequency of ingestion, as well as the oral hygiene practiced by the individual.

Nature, 2003 Nov 6, 426(6962), 33 - 8
The immune response of Drosophila; Hoffmann JA; Drosophila mounts a potent host defence when challenged by various microorganisms . Analysis of this defence by molecular genetics has now provided a global picture of the mechanisms by which this insect senses infection, discriminates between various classes of microorganisms and induces the production of effector molecules, among which antimicrobial peptides are prominent . An unexpected result of these studies was the discovery that most of the genes involved in the Drosophila host defence are homologous or very similar to genes implicated in mammalian innate immune defences . Recent progress in research on Drosophila immune defence provides evidence for similarities and differences between Drosophila immune responses and mammalian innate immunity.

Appl Environ Microbiol, 2003 Nov, 69(11), 6777 - 84
Binding of pediocin PA-1 with anionic lipid induces model membrane destabilization; Gaussier H et al.; To obtain molecular insights into the action mode of antimicrobial activity of pediocin PA-1, the interactions between this bacteriocin and dimyristoylphosphatidylcholine (DMPC) or dimyristoylphosphatidylglycerol (DMPG) model membranes have been investigated in D(2)O at pD 6 by Fourier transform infrared spectroscopy . The interactions were monitored with respect to alteration of the secondary structure of pediocin, as registered by the amide I' band, and phospholipid conformation, as revealed by the methylene nu(s)(CH(2)) and carbonyl nu(C;O) stretching vibrations . The results show that no interaction between pediocin and DMPC occurs . By contrast, pediocin undergoes a structural reorganization in the presence of DMPG . Upon heating, pediocin self-aggregates, which is not observed for this pD in aqueous solution . The gel-to-crystalline phase transition of DMPG shifts to higher temperatures with a concomitant dehydration of the interfacial region . Our results indicate that pediocin is an extrinsic peptide and that its action mechanism may lie in a destabilization of the cell membrane.

Int J Antimicrob Agents, 2003 Nov, 22(5), 479 - 86
Quinoline and cyanine dyes--putative anti-MRSA drugs; Wainwright M et al.; One way in which drug-resistant bacteria may be attacked is to screen new series of candidate compounds . Quaternary quinoline compounds and dyes were studied by Carl Browning (1887-1972) and Julius Cohen (1859-1935) . A remarkable part of Browning and Cohen's work was the early development of structure-activity relationships for their series of compounds . Thus cationic species were found generally to be more effective antibacterials than neutrals or anionics, and the testing of partial or deconstructed active molecules was also carried out . Much of this work underpinned the fuller understanding of e.g . aminoacridine action developed by Adrien Albert (1907-1989), himself also a collaborator of Browning . Analysis of the activity of a range of compounds developed by Browning and Cohen suggests that these might again be examined as topical antimicrobials in the fight against methicillin-resistant S . aureus (MRSA) and other resistant bacteria.

Int J Antimicrob Agents, 2003 Nov, 22(5), 465 - 78
Antimicrobial peptides in animals and their role in host defences; Brogden KA et al.; Domesticated animals have a large variety of antimicrobial peptides that serve as natural innate barriers limiting microbial infection or, in some instances, act as an integral component in response to inflammation or microbial infection . These peptides differ in size, composition, mechanisms of activity and range of antimicrobial specificities . They are expressed in many tissues, polymorphonuclear leukocytes, macrophages and mucosal epithelial cells . There is a small group of anionic antimicrobial peptides found in ruminants and a much larger group of cationic antimicrobial peptides found in all domesticated animals . The cationic peptides include linear, helical peptides, linear peptides rich in proline and cysteine-stabilized peptides with a beta-sheet and are commonly referred to as cathelicidins and defensins . These peptides are generally broad-spectrum for Gram-positive bacteria, Gram-negative bacteria and fungi (e.g . myeloid antimicrobial peptides, alpha-, beta-defensins, and protegrins) or are specific to one of these groups (e.g . porcine cecropin P1, Bac5, Bac7, PR-39 and prophenin).

Comp Biochem Physiol B Biochem Mol Biol, 2003 Nov, 136(3), 505 - 13
Antibacterial properties of serum from the American alligator (Alligator mississippiensis); Merchant ME et al.; Treatment of alligator (Alligator mississippiensis) and human serum samples with Escherichia coli resulted in a time- and concentration-dependent inhibition of bacterial proliferation . When inoculated with E . coli, alligator serum exhibited 10-fold lower bacterial survival rates after 1 h than human serum . In addition, the antibacterial spectrum of alligator serum was shown to be much broader than that of human serum, with growth inhibition occurring in 100% of bacterial strains tested (compared to only 35% for human serum) . Additional results showed that the antimicrobial activities of alligator serum could be completely inhibited by preincubation with proteases, indicating the proteinaceous nature of the antimicrobial activities . Furthermore, incubation of alligator serum at 56 degrees C for 30 min (classical human serum complement inactivation conditions) obliterated all antimicrobial properties of the alligator serum . The antibacterial activities occurred relatively quickly in vitro, with significant activity occurring within 5 min of inoculation with E . coli and maximal activity at 20 min . Also, the antimicrobial activity exhibited temperature dependence, with a substantial decrease in activity below 15 degrees C . These data suggest that the antimicrobial properties of alligator serum may be due to an active serum complement system.

Mayo Clin Proc, 2003 Nov, 78(11), 1409 - 11
Ocular ethambutol toxicity; Melamud A et al.; Ethambutol is an antimicrobial agent used frequently to treat tuberculosis . The most commonly recognized toxic effect of ethambutol is optic neuropathy, which generally is considered uncommon and reversible in medical literature . We describe a 43-year-old man who developed signs and symptoms of bilateral optic neuropathy during treatment with ethambutol . This case and a review of the literature show the severe and unpredictable nature of ethambutol toxicity and its potential for irreversible vision loss despite careful ophthalmologic monitoring.

Transgenic Res, 2003 Oct, 12(5), 597 - 605
Expression of lysostaphin in milk of transgenic mice affects the growth of neonates; Mitra A et al.; As an initial step towards enhancing mastitis resistance in dairy animals, we generated BLG-Lys transgenic mice that secrete lysostaphin, a potent antistaphylococcal protein, in their milk . In the current study, we continue our assessment of lysostaphin as a suitable antimicrobial protein for mastitis resistance and have investigated mammary gland development and function in three lines of transgenic mice . As the lines were propagated, there was a tendency for fewer BLG-Lys litters to survive to weaning (51% as compared to 90% for nontransgenic lines, p = 0.080) . Nontransgenic pups fostered on dams from these three lines exhibited diminished growth rates during the first week of lactation . Rates of gain became comparable to pups on nontransgenic dams at later time points . Initial slow growth also resulted in decreased weaning weights for pups nursed by transgenic dams (15.35 +/- 0.27 g) when compared to pups delivered and nursed by nontransgenic dams (18.61 +/- 0.61 g; p < 0.001), but the effect was temporary, as similar weights were attained by adulthood . Milk yield at peak lactation was not different between BLG-Lys (0.79 +/- 0.33 g) and nontransgenic (0.91 +/- 0.38 g; p = 0.166) dams . Histological examination of the transgenic mammary glands during gestation revealed no differences when compared to control glands; however, at early lactational stages, the BLG-Lys glands exhibited less alveolar area than control glands and a delay in lobulo-alveolar maturation . The results clearly demonstrate reduced growth of neonates on BLG-Lys dams; whether the poor pup performance can be attributed to delayed mammary development or the gland development merely reflects reduced suckling stimuli from the pups remains to be determined.

Microsc Res Tech, 2003 Dec 1, 62(5), 423 - 30
Transmission electron microscopic observations of membrane effects of antibiotic cecropin B on Escherichia coli; Chen HM et al.; The pathway of cell membrane lysis by the peptide antibiotic cecropin B (CB), which contains both a hydrophobic and an amphipathic alpha-helix, was analysed by assessing the morphological changes of Escherichia coli following treatment with the peptide . Exposure of green fluorescent protein (GFP)-expressing E . coli to CB does not lead to an efflux of GFP . Moreover, transmission electron microscopic (TEM) examination of cecropin B-treated cells showed that severe swelling precedes cell death and that the outer membrane becomes distended away from the plasma membrane . Using immuno-gold staining and TEM of E . coli expressing the maltose-binding protein in the cytoplasm, it was apparent that the protein remains restricted to the cytoplasmic compartment . These observations suggest that CB causes gross disruption of the outer membrane of Gram-negative bacteria . Circular dichroism measurements of CB in the presence of cell membrane-mimicking liposomes showed that CB forms secondary structure dependent on the ratio of {lipid}/{peptide} . These observations from this study are important for the future design of custom antimicrobial peptides .

Intensive Care Med, 2003 Dec, 29(12), 2327 - 9 Epub 2003 Nov 05.
Disseminated tuberculosis, pulmonary aspergillosis and cutaneous herpes simplex infection in a patient with infliximab and methotrexate; van der Klooster JM et al.; CASE PRESENTATION: Despite chemoprophylaxis with isoniazid a 58-year-old Creole patient with mild rheumatoid arthritis developed disseminated tuberculosis, pulmonary aspergillosis and cutaneous herpes simplex infection during treatment with infliximab and methotrexate . TREATMENT: The patient received antituberculous drugs (ethambutol, isoniazid, pyrazinamide, rifampicin), amphotericin B, flucytosine, and valaciclovir, along with prolonged intensive care treatment and mechanical ventilation . CONCLUSIONS: The present case confirms that isoniazid prophylaxis (300 mg once daily, during 6 months) does not protect against the reactivation and dissemination of latent tuberculosis . It also shows that combined treatment with infliximab and methotrexate may induce severe immunosuppression with prolonged leukocytopenia and depressed cellular immunity, leading to multiple opportunistic infections . Extensive diagnostic testing, early start of antimicrobial therapy and enteral immunonutrition, and further infection prevention with selective decontamination of the digestive tract may have been the key to a good clinical outcome.

Microbiology, 2003 Nov, 149(Pt 11), 3139 - 53
The 64 508 bp IncP-1beta antibiotic multiresistance plasmid pB10 isolated from a waste-water treatment plant provides evidence for recombination between members of different branches of the IncP-1beta group; Schluter A et al.; The complete 64508 bp nucleotide sequence of the IncP-1beta antibiotic-resistance plasmid pB10, which was isolated from a waste-water treatment plant in Germany and mediates resistance against the antimicrobial agents amoxicillin, streptomycin, sulfonamides and tetracycline and against mercury ions, was determined and analysed . A typical class 1 integron with completely conserved 5' and 3' segments is inserted between the tra and trb regions . The two mobile gene cassettes of this integron encode a beta-lactamase of the oxacillin-hydrolysing type (Oxa-2) and a gene product of unknown function (OrfE-like), respectively . The pB10-specific gene load present between the replication module (trfA1) and the origin of vegetative replication (oriV) is composed of four class II (Tn3 family) transposable elements: (i) . a Tn501-like mercury-resistance (mer) transposon downstream of the trfA1 gene, (ii) . a truncated derivative of the widespread streptomycin-resistance transposon Tn5393c, (iii) . the insertion sequence element IS1071 and (iv) . a Tn1721-like transposon that contains the tetracycline-resistance genes tetA and tetR . A very similar Tn501-like mer transposon is present in the same target site of the IncP-1beta degradative plasmid pJP4 and the IncP-1beta resistance plasmid R906, suggesting that pB10, R906 and pJP4 are derivatives of a common ancestor . Interestingly, large parts of the predicted pB10 restriction map, except for the tetracycline-resistance determinant, are identical to that of R906 . It thus appears that plasmid pB10 acquired as many as five resistance genes via three transposons and one integron, which it may rapidly spread among bacterial populations given its high promiscuity . Comparison of the pB10 backbone DNA sequences with those of other sequenced IncP-1beta plasmids reveals a mosaic structure . While the conjugative transfer modules (trb and tra regions) and the replication module are very closely related to the corresponding segments of the IncP-1beta resistance plasmid R751 and even more similar to the IncP-1beta degradative plasmids pTSA and pADP-1, the stable inheritance operons klcAB-korC and kleAEF are most similar to those of the IncP-1beta resistance plasmid pB4, and clearly less similar to the other IncP-1beta plasmids . This suggests that IncP-1beta plasmids can undergo recombination in the environment, which may enhance plasmid diversity and bacterial adaptability.

Am J Public Health, 2003 Nov, 93(11), 1910 - 4
Trends in antimicrobial prescribing for bronchitis and upper respiratory infections among adults and children; Mainous AG 3rd et al.; OBJECTIVES: This study examined antimicrobial prescribing patterns for adults and children with bronchitis or upper respiratory infections (URIs) before and after release of nationally disseminated pediatric practice recommendations . METHODS: Data from the 1993, 1995, 1997, and 1999 National Ambulatory Medical Care Survey were used to evaluate prescriptions for antimicrobials for URIs and bronchitis . RESULTS: From 1993 to 1999, the proportion of children receiving antimicrobials after visits for URIs and bronchitis decreased . However, the use of broad-spectrum antimicrobials rose from 10.6% of bronchitis visits to 40.5% . Prescriptions of antimicrobials for adults with URIs or bronchitis showed a decrease between 1993 and 1999 . CONCLUSIONS: Although antimicrobial prescribing for URIs and bronchitis has decreased for both children and adults, the prescribing of broad-spectrum antibiotics among children has shown a proportional rise.

Regul Pept, 2003 Nov 15, 116(1-3), 139 - 46
Identification of three novel Phyllomedusa sauvagei dermaseptins (sVI-sVIII) by cloning from a skin secretion-derived cDNA library; Chen T et al.; The defensive skin secretions of many amphibians contain a wide spectrum of biologically active compounds, particularly antimicrobial peptides that act as a first line of defence against bacterial infection . Here we describe for the first time the identification of three novel dermaseptin-related peptides (dermaseptins sVI-sVIII) whose primary structures were deduced from cDNAs cloned from a library constructed from lyophilised skin secretion of the South American hylid frog, Phyllomedusa sauvagei . The molecular masses of each were subsequently confirmed by interrogation of archived LC/MS files of fractionated skin secretion followed by automated Edman degradation sequencing . The heterogeneity of primary structures encountered in amphibian skin antimicrobial peptides may in part be explained by individual variation-a factor essential for selective functional molecular evolution and perhaps, ultimately in speciation.

Am J Med, 2003 Nov, 115(7), 529 - 35
Inappropriate initial antimicrobial therapy and its effect on survival in a clinical trial of immunomodulating therapy for severe sepsis; Harbarth S et al.; PURPOSE: To examine the effect of inappropriate initial antimicrobial therapy on the prognosis of patients with sepsis who were enrolled in a clinical trial of an immunomodulating agent conducted in 108 hospitals in North America and Europe . METHODS: We assessed initial antimicrobial choice and results of microbiologic cultures in 904 patients who had microbiologically confirmed severe sepsis or early septic shock . If a patient did not receive at least one antimicrobial agent to which the causative microorganisms were susceptible within 24 hours from the diagnosis of severe sepsis, then the initial antimicrobial treatment was considered to be inappropriate . A propensity score that adjusted for factors associated with inappropriate antimicrobial treatment was calculated and included in multivariable models to adjust for confounding . RESULTS: A total of 468 patients (52%) had documented bloodstream infection, and 211 patients (23%) received inappropriate initial antimicrobial therapy . Characteristics associated with inappropriate treatment were study enrollment in Europe, admission to surgery, nosocomial infection, infection with multiresistant microorganisms, and fungal or polymicrobial infection (all P <0.05) . The 28-day mortality was 24% (168/693) for patients in the adequately treated group versus 39% (82/211) for patients receiving inappropriate initial antimicrobial therapy (P <0.001) . After adjusting for comorbid conditions, severity of illness, site of infection, and the propensity score, inappropriate antimicrobial therapy was independently associated with increased mortality (odds ratio = 1.8; 95% confidence interval: 1.2 to 2.6) . CONCLUSION: In a large cohort of patients with microbiologically confirmed severe sepsis, appropriate initial antimicrobial therapy was an important determinant of survival . New approaches aimed at improving detection and treatment of early sepsis are needed.

Int J Tuberc Lung Dis, 2003 Nov, 7(11), 1027 - 32
The potential of human neutrophil peptides in tuberculosis therapy; Fu LM; The problems of drug resistance and bacterial persistence in tuberculosis have prompted scientists to search for clues from the latest advances in microbiology and immunology . Recent research on human neutrophil peptides (HNPs) has highlighted their bactericidal action against Mycobacterium tuberculosis and suggested that neutrophils may play a more important defensive role in tuberculosis than previously thought . Human neutrophil peptides belong to a family of antimicrobial and cytotoxic peptides known as 'defensins' . Neutrophils use both oxidative and non-oxidative microbicidal mechanisms to provide the host with innate immunity against microbial infections . Defensins are most abundant among an array of oxygen-independent antimicrobial proteins and peptides in neutrophil granules . Defensins are effective against a wide spectrum of microbes including bacteria, viruses, fungi, spirochetes and mycobacteria . In addition to direct antimicrobial activity, HNPs can potentially influence the inflammatory or immune responses by modulating cytokine production or acting like opsonins or chemotactic factors . HNPs are active against M . tuberculosis grown in vitro or within macrophages . HNPs released by neutrophils recruited in the early lesion could attract monocytes to the site and macrophages may in vivo uptake the extracellular HNPs and kill the intracellular pathogens . As such, HNPs are potential therapeutic agents against tuberculosis . HNPs are also cytotoxic against a wide range of normal mammalian cells; however, there is evidence that defensins may not cause significant cytotoxicity at the therapeutic level . Finally, the clinical application of HNPs must be evaluated in the context of possible drug resistance, as some resistance-associated genes have been identified.

J Chemother, 2003 Oct, 15(5), 466 - 71
Comparison of five antimicrobial regimens for the treatment of brucellar spondylitis: a prospective, randomized study; Bayindir Y et al.; Brucellosis, a zoonosis with worldwide distribution, is a systemic infection and still an important public health problem in Turkey . The best antimicrobial combination and schedule for the treatment of brucellosis with spondylitis has not yet been clearly determined . In a prospective and randomized study, we compared the efficacy of five antimicrobial regimens for treatment of 102 patients with lumbar brucellar spondylitis . Patients were randomly assigned to receive antimicrobial combination therapy . Twenty patients received streptomycin 1 g/day intramuscularly for 15 days and tetracycline-HCl, 500 mg every 6 h orally for 45 days (ST), 21 patients received streptomycin 1 g/day i.m . for 15 days and doxycycline 100 mg every 12 h orally for 45 days (SD), 20 patients received doxycycline 100 mg every 12 h orally for 45 days and rifampicin 15 mg/kg per day in a single morning dose orally for 45 days (DR), 19 patients received ofloxacin, 200 mg every 12 h orally for 45 days and rifampicin 15 mg/kg per day in a single morning dose orally for 45 days (OR), and 22 patients received streptomycin 1 g/day i.m . for 15 days and doxycycline 100 mg every 12 h orally for 45 days plus rifampicin 15 mg/kg per day in a single morning dose orally for 45 days (SDR) . Initial therapeutic failure occurred in 2 patients (10%) in the ST regimen group, 4 patients (19%) in the SD group, 3 patients (15%) in the DR group and 5 patients (26%) in the OR regimen . In addition, 2 patients (10%) in the DR group and 5 patients (26%) in the OR regimen relapsed during the follow-up period . There was no relapse in any patients in the ST, SD, and SDR groups . The response rates were 90% in the ST and 81% in the SD groups . In contrast, there was a maximum good response (100%) and no relapse in the SDR group . In conclusion, a combination of doxycycline, streptomycin, and rifampicin can be recommended as therapy for brucellar spondylitis and to reduce relapse rates.

J Clin Invest, 2003 Nov, 112(9), 1300 - 7
Pharmacological inhibition of quorum sensing for the treatment of chronic bacterial infections; Hentzer M et al.; Traditional treatment of infectious diseases is based on compounds that aim to kill or inhibit bacterial growth . A major concern with this approach is the frequently observed development of resistance to antimicrobial compounds . The discovery of bacterial-communication systems (quorum-sensing systems), which orchestrate important temporal events during the infection process, has afforded a novel opportunity to ameliorate bacterial infection by means other than growth inhibition . Compounds able to override bacterial signaling are present in nature . Herein we discuss the known signaling mechanisms and potential antipathogenic drugs that specifically target quorum-sensing systems in a manner unlikely to pose a selective pressure for the development of resistant mutants.

Gene, 2003 Nov 13, 319, 43 - 53
Characterization and transcriptional profiles of three Spodoptera frugiperda genes encoding cysteine-rich peptides . A new class of defensin-like genes from lepidopteran insects?
Volkoff AN, Rocher J, d'Alencon E, Bouton M, Landais I, Quesada-Moraga E, Vey A, Fournier P, Mita K, Devauchelle G.
The present work describes sequence and transcription of three Spodoptera frugiperda genes encoding 6-cysteine-rich peptides . Sequence alignments indicate that the predicted peptides belong to the insect defensin family, although phylogenetic analyses suggest they form a cluster distinct from that of other neopteran insect defensins . The three genes were identified in a non-immune-challenged Sf9 cells cDNA (DNA complementary to RNA) library (Landais et al., Bioinformatics, in press) and were named spodoptericin, Sf-gallerimycin and Sf-cobatoxin . Spodoptericin is a novel defensin-like gene that appears to be weakly up-regulated following injection of bacteria and fungi . Interestingly, no sequence motif clearly homologous to cis regulatory element involved in the regulation of antimicrobial genes was found . An homologue of the spodoptericin gene was identified in the SilkBase Bombyx mori cDNA library . Sf-gallerimycin is related to the Galleria mellonella gallerimycin gene and is induced after immune challenge by injection of bacteria in the larval fat body as well as in hemocytes . In silico analysis of the sequence upstream from the cDNA reveals the presence of at least one motif homologous to a nuclear factor kappaB (NF-kappaB) binding site . Finally, Sf-cobatoxin is related to the G . mellonella cobatoxin-like gene . Despite high levels of constitutive expression compared to the two previous genes, transcription of Sf-cobatoxin is increased after immune, in particular, bacterial challenge . We therefore confirm that these three genes encode potential candidate molecules involved in S . frugiperda innate humoral response.

FEBS Lett, 2003 Nov 6, 554(1-2), 100 - 4
Individual substitution analogs of Mel(12-26), melittin's C-terminal 15-residue peptide: their antimicrobial and hemolytic actions; Yan H et al.; Residues 1-9 of M(12-26) (GLPALISWIKRKRQQ-NH2), the C-terminal 15-residue segment of melittin, were substituted individually to change the hydropathicities in these positions . Antimicrobial and hemolytic activities of these peptides were determined . The results showed increased antimicrobial activities with increased hydrophobicities at almost all the positions studied . The effects at positions 2, 5, 8 and 9 were significant while the effects at the other positions were small . These two groups of residues were located on the opposite faces of the alpha-helix . In other words, the hydrophobicities of the two faces were favorable, but one face (the more favorable face) contributed more to the antimicrobial activities than the other (the less favorable face) . The hydrophobicity, not the amphipathicity, seems to be crucial for antimicrobial activity . In contrast, the hydrophobicity of one face was favorable but the other was unfavorable for the hemolytic activity, indicating that the amphipathicity may be important for hemolysis . Interestingly, the more favorable face for antimicrobial activity was located opposite to the favorable face for hemolytic activity, indicating the direction of the hydrophobic face for the antimicrobial activity and direction of the amphipathicity for the hemolytic activity were also important.

Biochemistry, 2003 Nov 11, 42(44), 12866 - 74
Analysis of membrane-binding properties of dermaseptin analogues: relationships between binding and cytotoxicity; Gaidukov L et al.; To understand relationships between membrane-binding properties of cytolytic peptides and resulting cytotoxicity, we investigated interactions of dermaseptin analogues with model bilayers by means of surface plasmon resonance . First, we tested the system by comparing two native dermaseptins, S1 and S4, whose binding properties were previously characterized in different experimental systems . Validation experiments revealed deviations from the one-to-one interaction model and indicated the binding to proceed by a two-stage mechanism . By calculation of apparent affinity constants and individual affinities for both steps of the interaction, the biosensor technology was able to distinguish between surface-bound peptides that subsequently penetrated into the bilayer and peptides that remained essentially superficially bound . This data interpretation was sustained after analysis of a series of dermaseptin S4 derivatives whose binding data were compared with cytotoxicity, revealing cytolytic activity to correlate mainly with insertion affinity . The data indicate that the potency of highly cytolytic peptides such as K(4)K(20)-S4 is not due to the highest membrane adhesion affinity but to the highest propensity for the inserted state . Similarly, truncated derivatives of 16, 13, and 10 residues showed a progressive reduction in cytotoxicity that best correlated with progressive reduction in insertion affinity . Support for the adhesion versus inserted states was provided by proteolytic experiments with RBC-bound peptides that demonstrated K(4)K(20)-S4 to be protected from enzymatic cleavage, unlike its 13-mer derivative . Overall, using the two-stage model proved instrumental in investigating membrane-binding properties of antimicrobial peptides and capable of explaining the cytolytic properties of closely related analogues.

Phytother Res, 2003 Nov, 17(9), 1082 - 7
Biological activities of Prunella vulgaris extract; Psotova J et al.; The organic fraction (OF; 25.7% w/w of rosmarinic acid) of Prunella vulgaris (total extract) was found to exhibit the following: scavenging activity on diphenylpicrylhydrazyl radical (DPPH), inhibition of in vitro human LDL Cu(II)-mediated oxidation, protection of rat mitochondria and rat hepatocytes exposed to either tert-butyl hydroperoxide, or to Cu(II) and Fe(III) ions . OF also showed a potential to inhibit rat erythrocyte haemolysis and it reduced the production of LTB(4) in bovine PMNL generated by the 5-lipoxygenase pathway . Other observations included antiproliferative effects against HaCaT cells and mouse epidermal fibroblasts and a moderate OF antimicrobial activity on gram-positive bacteria . Rosmarinic, caffeic and 3-(3,4-dihydroxyphenyl)lactic acids exhibited less potent activity than the plant extract in all bioassays . The antioxidative, antimicrobial, together with antiviral effects offer good prospects for the medicinal applications of P . vulgaris .

J Infect Dis, 2003 Nov 1, 188(9), 1382 - 93 Epub 2003 Oct 20.
Protective efficacy of CAP18106-138-immunoglobulin G in sepsis; Warren HS et al.; Naturally present antibacterial proteins play an important role in innate host defense . A synthetic peptide mimicking the C-terminal lipopolysaccharide (LPS)-binding domain of rabbit cathelicidin CAP18 was coupled to immunoglobulin (Ig) G to create CAP18(106-138)-IgG, a construct that, in concentrations equimolar to those of peptide alone, binds and neutralizes LPS and kills multiple gram-negative bacterial strains . The protective efficacy of CAP18(106-138)-IgG was evaluated in a model of cecal ligation and puncture in mice . A single intravenous administration of 20 mg/kg CAP18(106-138)-IgG protected against mortality, compared with sham-coupled IgG (P<.03) . There was no protection offered by administration of equimolar peptide alone (P=.96) . There was a trend toward protection in C3H/HeJ mice that are minimally sensitive to LPS (P=.06), suggesting that direct detoxification of LPS was not the only mechanism of protection . Chemical or genetic coupling of antimicrobial peptides to IgG may be a means of using these peptides to treat infections.

Bioorg Med Chem Lett, 2003 Nov 17, 13(22), 3993 - 6
Rational design of antimicrobial agents: antifungal activity of alk(en)yl dihydroxybenzoates and dihydroxyphenyl alkanoates; Nihei K et al.; A homologous series (C3-C14) of each alkyl 3,4- and 3,5-dihydroxybenzoates, and 3,4- and 3,5-dihydroxyphenyl alkanoates exhibit similar antifungal activity against Saccharomyces cerevisiae . Their nonyl derivatives exhibit the most potent antifungal activity against this yeast with the minimum fungicidal concentration (MFC) in the range between 12.5 and 50 microg/mL . In addition, various 3,4-dihydroxybenzoates, possessing different side chains, namely unsaturated, branched and alicyclic were synthesized and their activity was compared.

Surg Infect (Larchmt), 2003 Fall, 4(3), 273 - 80
Prophylaxis of infection for elective colorectal surgery; Jimenez JC et al.; BACKGROUND: Influenced by the key results of the clinical trials conducted in the early 1970s by Condon, Nichols, and Gorbach, surgeons have adopted the routine use of mechanical bowel prep and antimicrobial prophylaxis prior to elective colorectal procedures as a widely established practice . Recent clinical trial data, however, led us to reexamine the benefits of mechanical bowel preparation, methods of antimicrobial prophylaxis and to assess the role of new, specific risk factors for surgical site infection after colorectal operations . METHODS: Pertinent studies on antimicrobial prophylaxis for elective colorectal surgery were identified from a Medline search of English language publications since 1966 . RESULTS: We found credible clinical trial data that mechanical bowel preparation prior to elective colorectal surgery may not be essential . Timing of the administration of prophylactic antimicrobials is often inaccurate in current practice and suggests the need for a long-acting, broad-spectrum agent that would deemphasize precision in time of preoperative infusion . New risk factors have been identified that increase infection after colorectal surgery, including patient core temperature and tissue oxygenation . Independent observers identify postoperative surgical site infection at a higher rate than physician self-reporting and should be incorporated into future clinical trials . CONCLUSION: The once settled area of antimicrobial prophylaxis for colorectal surgery is again controversial . Cooperative clinical trials will be needed to resolve key questions such as the efficacy for bowel preparation and how to obtain effective timing of antimicrobial prophylaxis.

Infect Control Hosp Epidemiol, 2003 Oct, 24(10), 782 - 4
Light-activated antimicrobial coating for the continuous disinfection of surfaces; Wilson M; The ability of a photosensitizer-containing cellulose acetate film to kill bacteria was evaluated . Substantial kills were achieved following irradiation of the film with white light for up to 24 hours . Applying a photosensitizer-containing coating to surfaces could reduce the environmental load of pathogens, thus helping to prevent infectious disease transmission.

Infect Control Hosp Epidemiol, 2003 Oct, 24(10), 762 - 4
Antimicrobial activity of glucoprotamin: a clinical study of a new disinfectant for instruments; Widmer AE et al.; OBJECTIVE: To determine the in vitro efficacy of glucoprotamin for the disinfection of instruments . DESIGN: Prospective observational study . SETTING: University women's hospital . METHODS: Instruments were immersed in saline solution after use, and glucoprotamin was added to a concentration of 1.5% before soaking for 60 minutes . Biocidal activity was determined by the difference in colony-forming units (CFU) on instruments before and after disinfection . RESULTS: One hundred thirty-seven instruments were collected during 10 days and exposed to a 1.5% dilution of glucoprotamin without prior washing . Bioburden before disinfection ranged from 2 x 10(5) to 7.1 x 10(7) CFU per instrument . Average bacterial killing was 5.98 log10 CFU +/- 0.48 under aerobic conditions and 6.75 log10 CFU +/- 0.54 under anaerobic conditions, despite the presence of large amounts of proteins on instruments that were frequently bloody . No vegetative bacteria were isolated in any sample after disinfection . CONCLUSION: This clinical study confirmed excellent in vitro efficacy of glucoprotamin without prior removal of proteins and debris.

Otolaryngol Pol, 2003, 57(4), 581 - 6
{Bezold's abscess: a rare complication of otitis media}; Steczko A et al.; The authors present the case of a 21-year-old female in who, during the course of cholesteatoma, extracranial complications occurred twice in the form of Bezold's abscess . This type of the complication of otitis media occurs extremely rare . Medical literature has recorded about 20 cases of such complications . Due to commonly applied antibiotic therapies, extracranial and intracranial complications are rarely recorded in the course of otitis media, their mortality rate, however, amounts to a dozen or so per cent . The paper presents the way in witch the infection of otitis media spreads towards the neck . The authors stress the significance of computer tomography in diagnostic . Radical mastoidectomy and exploration of the neck were performed on the patient . Antibiotic regimens were based on antimicrobial sensitivity patterns . Presently, the patients is under constant laryngologist observation.

Cell Tissue Res, 2004 Jan, 315(1), 59 - 70 Epub 2003 Oct 28.
Cutaneous eccrine glands of the foot pads of the small Madagascan tenrec ( Echinops telfairi, Insectivora, Tenrecidae): skin glands in a primitive mammal; Stumpf P et al.; In order to find correlations between skin gland morphology and specific ethological features, the cutaneous glands of the foot pads of the primitive mammal the Madagascan tenrec, Echinops telfairi, were studied by histological and various histochemical methods as well as by electron microscopy . In the foot pads specific eccrine skin glands occurred consisting of coiled ducts and tubular secretory portions, the lumina of which were considerably wider than in primate sweat glands . The secretory tubules were composed of branched myoepithelial cells and glandular cells . The latter contained abundant mitochondria, large amounts of glycogen particles and few secretory granules as well as individual heterolysosomes and myelin bodies . The lateral cell membrane was marked by extensive interdigitations . The apical membranes of all glandular cells contained proteoglycans with sulfated and carboxylated groups containing N-acetyl-glucosamine, N-acetyl-galactosamine, galactose and mannose . The expression pattern of cytokeratins of the glandular epithelium was variable and showed similarities to that of the human eccrine glands . Tubulin, vinculin and actin were expressed in the glandular epithelium . The secretory cells showed positive reactions with antibodies against antimicrobial peptides and IgA . A positive reaction was observed with antibodies against the androgen receptor . The PCNA and TUNEL reactions indicated that the tubular skin glands of Echinops are made up of a slowly renewing tissue . We conclude that the glands fulfill several functions: production of a fluid-rich secretory product, which may prevent slipping of the foot pads on the substrate during running or climbing, secretion of antimicrobial peptides and proteins, and playing a role in thermoregulation.

Drugs Today (Barc), 2003 Sep, 39(9), 733 - 8
Cycling chemotherapy: a promising approach to reducing the morbidity and mortality of nosocomial infections; Evans HL et al.; In the past several decades, nosocomial infections have emerged as one of the most serious contributors to hospital morbidity and mortality, particularly amongst patients who require intensive care . Resistant organisms, both Gram-negative and Gram-positive, are now to blame for a significant portion of hospital-acquired infections . Efforts to prevent nosocomial infection had historically focused on infection control measures, such as patient isolation . However, there have been numerous reports of the increasing prevalence of antibiotic resistance, as well as the dramatic, negative impact of the infections they cause, both in terms of patient outcomes and attributable costs, demanding new methods to halt this growing epidemic . The increasing threat of resistance may be attributed in part to the widespread and increasingly inappropriate use of antimicrobials, which inadvertently exert sufficient effect on the hospital (and now community) environment to allow the preferential selection of resistant microbes . This idea of selective antibiotic pressure is supported by data showing volume of antibiotic use and inadequate antimicrobial coverage as risk factors for increased morbidity and mortality . Accordingly, the focus of nosocomial infection control has now largely shifted towards the judicious use of antibiotic therapy . There have been numerous attempts to curtail antibiotic usage through various forms of antibiotic stewardship: formulary restriction, computerized decision-support and abbreviated course empiric therapy . Aside from the inherent difficulty of effecting change in physician practice, we are burdened, particularly in the setting of empiric therapy, with the need to balance between adequate therapy for the individual and prudent drug selection so as not to endanger other patients in the environment through resistant organism selection . Cycling chemotherapy for empiric treatment of suspected infection is a method uniquely designed to address these challenges . (c) 2003 Prous Science . All rights reserved.

Mol Cell Biol, 2003 Nov, 23(22), 8272 - 81
Functional characterization of a novel promoter element required for an innate immune response in Drosophila; Uvell H et al.; Innate immune reactions are crucial processes of metazoans to protect the organism against overgrowth of faster replicating microorganisms . Drosophila melanogaster is a precious model for genetic and molecular studies of the innate immune system . In response to infection, the concerted action of a battery of antimicrobial peptides ensures efficient killing of the microbes . The induced gene expression relies on translocation of the Drosophila Rel transcription factors Relish, Dif, and Dorsal to the nucleus where they bind to kappaB-like motifs in the promoters of the inducible genes . We have identified another putative promoter element, called region 1 (R1), in a number of antimicrobial peptide genes . Site-directed mutagenesis of the R1 site diminished Cecropin A1 (CecA1) expression in transgenic Drosophila larvae and flies . Infection of flies induced a nuclear R1-binding activity that was unrelated to the kappaB-binding activity in the same extracts . Although the R1 motif was required for Rel protein-mediated CecA1 expression in cotransfection experiments, our data argue against it being a direct target for the Drosophila Rel proteins . We propose that the R1 and kappaB motifs are targets for distinct regulatory complexes that act in concert to promote high levels of antimicrobial peptide gene expression in response to infection.

Mol Cell Biol, 2003 Nov, 23(22), 7982 - 91
Discrete functions of TRAF1 and TRAF2 in Drosophila melanogaster mediated by c-Jun N-terminal kinase and NF-kappaB-dependent signaling pathways; Cha GH et al.; Two Drosophila tumor necrosis factor receptor-associated factors (TRAF), DTRAF1 and DTRAF2, are proposed to have similar functions with their mammalian counterparts as a signal mediator of cell surface receptors . However, their in vivo functions and related signaling pathways are not fully understood yet . Here, we show that DTRAF1 is an in vivo regulator of c-Jun N-terminal kinase (JNK) pathway in Drosophila melanogaster . Ectopic expression of DTRAF1 in the developing eye induced apoptosis, thereby causing a rough-eye phenotype . Further genetic interaction analyses revealed that the apoptosis in the eye imaginal disc and the abnormal eye morphogenesis induced by DTRAF1 are dependent on JNK and its upstream kinases, Hep and DTAK1 . In support of these results, DTRAF1-null mutant showed a remarkable reduction in JNK activity with an impaired development of imaginal discs and a defective formation of photosensory neuron arrays . In contrast, DTRAF2 was demonstrated as an upstream activator of nuclear factor-kappaB (NF-kappaB) . Ectopic expression of DTRAF2 induced nuclear translocation of two Drosophila NF-kappaBs, DIF and Relish, consequently activating the transcription of the antimicrobial peptide genes diptericin, diptericin-like protein, and drosomycin . Consistently, the null mutant of DTRAF2 showed immune deficiencies in which NF-kappaB nuclear translocation and antimicrobial gene transcription against microbial infection were severely impaired . Collectively, our findings demonstrate that DTRAF1 and DTRAF2 play pivotal roles in Drosophila development and innate immunity by differentially regulating the JNK- and the NF-kappaB-dependent signaling pathway, respectively.

Biomaterials, 2004 Jan, 25(2), 327 - 33
Bioactive glass as a drug delivery system of tetracycline and tetracycline associated with beta-cyclodextrin; Domingues ZR et al.; The aim of this study was to evaluate the physical-chemical properties, in vivo biocompatibility and antimicrobial activity of bioactive glasses (BG) used as a controlled release device for tetracycline hydrochloride and an inclusion complex formed by tetracycline and beta-cyclodextrin at 1:1 molar ratio . The BG as well as their compounds loaded with tetracycline (BT) and tetracycline:beta-cyclodextrin (BTC) were characterized by FTIR spectroscopy, X-ray powder diffraction, differential scanning calorimetry and by scanning electron microscopy and energy dispersive spectroscopy . The in vivo test was carried out with female mice split into three groups treated with bioactive glass either without drugs, or associated with tetracycline, or with tetracycline:beta-cyclodextrin by subcutaneous implantation . The histological examination of tissue at the site of implantation showed moderate inflammatory reactions in all groups after 72 h . The bacterial effect was tested on A . actinomycetemcomitans suspended in BHI broth, with or without bioactive particles . A considerable bacteriostatic activity was found with BT and BTC glasses, as compared to plain glass . The presence of cyclodextrin was important to slow down the release of tetracycline for a long period of time and it was verified that the presence of tetracycline or its inclusion complex, tetracycline:beta-cyclodextrin, did not affect the bioactivity of the glass.

Biomaterials, 2004 Feb, 25(3), 545 - 56
Backgrounds of antibiotic-loaded bone cement and prosthesis-related infection; Hendriks JG et al.; Antibiotic-loaded bone cement has been in use for over 30 years for the fixation of total joint arthroplasties, although its mechanism of action is still poorly understood . This review presents the backgrounds of bone cements, prosthesis-related infection and antibiotic-loaded bone cements . It is shown that antibiotic-loaded bone cement has a significant effect on bacteria, particularly in animal and clinical studies . However, recently, antimicrobial resistance among bacteria has been ascribed to the antibiotic-loaded bone cement . The unresolved issues both regarding the action of antibiotic-loaded bone cement and the nature of the antimicrobial resistance necessitate further research into the interaction of antibiotic-loaded bone cement and bacteria.

AIDS Res Hum Retroviruses, 2003 Oct, 19(10), 875 - 81
The theta-defensin, retrocyclin, inhibits HIV-1 entry; Munk C et al.; Retrocyclin is a circular antimicrobial 18-residue peptide encoded in the human genome by a theta-defensin pseudogene . In the human genome, the gene for retrocyclin is inactivated by an in-frame stop codon in its signal sequence but its mature coding sequence is intact . The peptide corresponding to the processed human retrocyclin, generated by solid phase peptide synthesis, inhibited replication of R5 and X4 strains of HIV-1 in human cells . Luciferase reporter virus and Vpr-BLaM entry assays were used to demonstrate that retrocyclin specifically blocked R5 and X4 HIV-1 replication at entry . Surface plasmon resonance demonstrated that retrocyclin bound to soluble CD4 and gp120, but gp120 cell-binding assays revealed that retrocyclin did not fully inhibit the binding of soluble CD4 to gp120 . A fluorescent retrocyclin congener localized in cell-surface patches either alone or colocalized with CD4, CXCR4, and CCR5 . In the aggregate, these results suggest that retrocyclin blocks an entry step in HIV-1 replication . Retrocyclin represents a new class of small molecule HIV-1 entry inhibitors.

J Periodontol, 2003 Sep, 74(9), 1316 - 9
Modulation of Porphyromonas gingivalis proteinase activity by suboptimal doses of antimicrobial agents; Grenier D et al.; BACKGROUND: Antimicrobial agents are sometimes used as adjuncts for the treatment of aggressive and refractory forms of periodontitis . In this study, we used a culture plate assay to investigate the effect of suboptimal doses of antimicrobial agents on proteinase activity of Porphyromonas gingivalis . METHODS: A culture plate assay using gelatin as the substrate, which allows a semiquantitative determination of proteinase activity, was developed . Suboptimal inhibitory concentrations of tetracycline, minocycline, doxycycline, metronidazole, penicillin G, or chlorhexidine were added to the medium, and proteolysis zones were determined following the growth of three strains of P . gingivalis . The effect of antimicrobials on outer membrane vesicle-associated gingipains also was determined . RESULTS: The gelatin plate assay was a convenient, simple procedure for investigating the effect of suboptimal inhibitory concentrations of antimicrobial agents on proteinases produced by P . gingivalis . The largest reduction (> 75%) in the proteolysis zones produced by three strains of P . gingivalis was obtained with minocycline . Tetracycline and doxycycline also reduced the proteolysis zones . A suboptimal inhibitory concentration of chlorhexidine increased the proteolysis zones by up to 70% . Metronidazole and penicillin G produced no noticeable effect . The suboptimal inhibitory concentrations of minocycline, tetracycline, and doxycyline did not reduce the activity of outer membrane vesicle-associated Arg- and Lys-gingipains . CONCLUSION: Results from this study suggest that sublethal concentrations of some antimicrobial agents in subgingival sites have the potential to affect the physiology of P . gingivalis, notably by increasing or decreasing the proteolytic activity of the bacteria.

J Chem Ecol, 2003 Sep, 29(9), 2049 - 71
Identification and quantitation of allelochemicals from the lichen Lethariella canariensis: phytotoxicity and antioxidative activity; Toledo Marante FJ et al.; Phytotoxicity-based extraction and fractionation were employed to separate allelochemicals contained in an extract of Lethariella canariensis . Twelve phenolic substances were isolated from the phytotoxic fraction "Letharal" of the thalli . These were identified by spectroscopic methods, physicochemical constants, and HPLC chemical correlation, and determined to be atranol (2), chloroatranol (3), hematommic acid (4), chlorohematommic acid (5), methyl hematommate (6), methyl chlorohematommate (7) (new compound), ethyl hematommate (8), ethyl chlorohematommate (9), methyl beta-orsellinate (10), atranorin (11), chloroatranorin (12), and (+)-usnic acid (13) . Further identification and quantification of these allelochemicals in the environment were conducted by HPLC . Several phenolic compounds showed moderate antimicrobial activity . The cytostatic activity of the polyphenols was investigated on U937 and HL-60 cells . All compounds were assayed, with the exception of 10 . The "Letharal" mixture decreased cell viability in both cell lines . Protection against lipid peroxidation was investigated using brain homogenates . Compounds 2, 3, 6, 8, 11, and Letharal decreased H2O2/Fe+2 induced lipid peroxidation in a concentration-dependent manner, while 10 and 13 were unable to protect tissue against oxidative stress.

Cochrane Database Syst Rev . 2003;(4):CD002037.
Three- or four- versus two-drug antiretroviral maintenance regimens for HIV infection; Rutherford GW et al.; BACKGROUND: Combination antiretroviral therapy administered to HIV-infected individuals has been shown to decrease viral replication, improve immunologic function and delay the progression of HIV infection . However, because patient adherence to complicated combination-therapy antiretroviral regimens is difficult and because of concerns regarding the cumulative toxicity of antiretroviral drugs, regimens that utilize fewer antiretroviral agents are desirable . OBJECTIVES: To compare the use three- or four- versus two-drug antiretroviral maintenance regimens following successful initial therapy for HIV infection . SEARCH STRATEGY: The following electronic databases were searched for relevant randomized trials or reviews: 1 . MEDLINE for the years 1982-May 2003 using the search terms human immunodeficiency virus, antiretroviral therapy, maintenance therapy, zidovudine, lamivudine, indinavir, stavudine, saquinivir, nelfinavir, didanosine, zalcitabine, ritonovir, AIDS, anti-HIV agents, HIV infection and HIV seropositivity . 2 . AIDSLINE for the years 1982- May 2003 using the search terms antiretroviral therapy, maintenance therapy, zidovudine, lamivudine, indinavir, stavudine, saquinivir, nelfinavir, didanosine, zalcitabine, ritonovir, anti-HIV agents . 3 . The Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effectiveness and the Cochrane Clinical Trials Register in the Cochrane Library, through May 2003 . 4 . AIDSTRIALS, a specialist registry of current and completed trials maintained by the U.S . National Library of Medicine through May 2003 . The abstracts of relevant conferences, including the International Conferences on AIDS, the Conference on Retroviruses and Opportunistic Infections, the Infectious Disease Society of America annual meeting and the Interscience Conference on Antimicrobial Agents and Chemotherapy, as indexed by AIDSLINE, were also reviewed . Reference lists of all review articles and primary articles identified were also searched . SELECTION CRITERIA: Randomized controlled trials in which HIV-infected adults who had successfully completed initial three- or four-drug antiretroviral therapy were randomized to maintenance therapy with three or four drugs or maintenance therapy with two drugs . Successful initial therapy was defined by a plasma viral load of less than 500 copies/ml . DATA COLLECTION AND ANALYSIS: Two reviewers assessed eligibility and trial quality . Attempts were made to contact the authors of the included abstract . Data on the number of patients experiencing loss of viral suppression were abstracted by two reviewers . The data were pooled, where appropriate, to yield odds ratios, using random effects models . MAIN RESULTS: Four trials were identified including three published studies and one abstract . Compared to three- or four-drug maintenance therapy, maintenance therapies including fewer drugs were associated with a higher risk of virologic failure (loss of HIV suppression to non-detectable levels) . Combining the results of all four studies yielded an odds ratio of 5.55 (95% confidence interval, 3.14 - 9.80) . Similar results were obtained when the one abstract was excluded (odds ratio, 5.48; 95% confidence interval, 2.82 - 10.65) . Performing subgroup analyses of studies using similar induction and maintenance regimens gave similar results . Maintenance regimens of zidovudine and lamivudine compared to maintenance regimens with zidovudine, lamivudine and indinavir, were associated with significantly higher rates of virologic failure (odds ratio, 4.57; 95% confidence interval, 1.80 - 11.58) . Similarly, maintenance regimens that discontinued one or more protease inhibitor after including them in induction therapy were also associated with a significantly higher risk of virologic failure (odds ratio, 6.15; 95% confidence interval, 3.40 -11.10) . REVIEWER'S CONCLUSIONS: Although it is desirable to reduce the number of antiretroviral drugs given in combination therapy for reasons of compliance and toxicity, maintenance regimens with fewer drugs are associated with significantly increased resistance and risk of loss of viral suppression . Successful initial therapy, as evidenced by suppression of viral load, should not be modified in the maintenance phase unless clinically necessary.

Clin Infect Dis, 2003 Nov 15, 37(10), 1304 - 12 Epub 2003 Oct 17.
Successful treatment of Balamuthia amoebic encephalitis: presentation of 2 cases; Deetz TR et al.; Case histories are presented of 2 individuals (a 5-year-old girl and 64-year-old man) who developed encephalitis caused by the free-living amoeba Balamuthia mandrillaris . Both individuals survived after diagnosis and initiation of effective antimicrobial therapy . Immunostaining for Balamuthia-specific antibody levels identified the causative agent of the infections . Antimicrobial therapy with flucytosine, pentamidine, fluconazole, sulfadiazine, and a macrolide antibiotic (azithromycin or clarithromycin) was initiated . Phenothiazines (thioridazine and trifluoperazine) were also used . Both patients recovered, and there was no evidence of recrudescence of the disease at 2 and 6 years after onset of symptoms . Awareness of Balamuthia as the causative agent of encephalitis and early initiation of antimicrobial therapy were critical to the recovery of both patients . Although optimal antimicrobial therapy for Balamuthia amoebic encephalitis has yet to be determined, the antimicrobials used in these 2 cases effectively controlled the disease . These 2 individuals are the only known survivors of this otherwise fatal type of amoebic encephalitis.

Free Radic Biol Med, 2003 Oct 1, 35(7), 814 - 25
Peroxidation and externalization of phosphatidylserine associated with release of cytochrome c from mitochondria; Jiang J et al.; Production of reactive oxygen species (ROS) during apoptosis is associated with peroxidation of phospholipids particularly of phosphatidylserine (PS) . The mechanism(s) underlying preferential PS oxidation are not well understood . We hypothesized that cytochrome c (cyt c) released from mitochondria into cytosol acts as a catalyst that utilizes ROS generated by disrupted mitochondrial electron transport for PS oxidation . Selectivity of PS oxidation is achieved via specific interactions of positively charged cyt c with negatively charged PS . To test the hypothesis we employed temporary transfection of Jurkat cells with a pro-apoptotic peptide, DP1, a conjugate consisting of a protein transduction domain, PTD-5, and an antimicrobial domain, KLA {(KLAKLAK)2}, known to selectively disrupt mitochondria . We report that treatment of Jurkat cells with DP1 yielded rapid and effective release of cyt c from mitochondria and its accumulation in cytosol accompanied by production of H2O2 . Remarkably, this resulted in selective peroxidation of PS while more abundant phospholipids such as phosphatidylcholine (PC) and phosphatidylethanolamine (PE) remained nonoxidized . Neither PTD-5 alone nor KLA alone exerted any effect on PS peroxidation . Redox catalytic involvement of cyt c in PS oxidation was further supported by our data demonstrating that: (i) specific interactions of cyt c with PS resulted in the formation of EPR-detectable protein-centered tyrosyl radicals of cyt c upon its interaction with H2O2 in the presence of PS-containing liposomes, and (ii) integration of cyt c into cytochrome c null (Cyt c -/-) cells or HL-60 cells specifically stimulates PS oxidation in the presence of H2O2 or t-BuOOH, respectively . We further demonstrated that DP1 elicited externalization of PS on the surface of Jurkat cells and enhanced their recognition and phagocytosis by J774A.1 macrophages . Our results are compatible with the hypothesis that catalysis of selective PS oxidation during apoptosis by cytosolic cyt c is important for PS-dependent signaling pathways such as PS externalization and recognition by macrophage receptors.

Spectrochim Acta A Mol Biomol Spectrosc, 2003 Nov, 59(13), 3111 - 22
E.s.r., magnetic, optical and biological (SOD and antimicrobial) studies of imidazolate bridged Cu(II)-Zn(II) and Cu(II)-Ni(II) complexes with tris(2-amino ethyl)amine as capping ligand: a plausible model for superoxide dismutase; Singh N et al.; X-band e.s.r . and optical absorption spectra of the imidazolate bridged heterobimetallic complexes {(tren)Cu-E-Im-Zn-(tren)}(ClO(4))(3) and {(tren)Cu-E-Im-Ni-(tren)}(ClO(4))(3), where trentris(2-aminoethyl)amine, E-Im=2-ethylimidazolate ion and the related mononuclear complexes {Cu(tren)}(ClO(4))(2) and {(tren)Cu-E-ImH)}(ClO(4))(2) have been described . Biological activities (superoxide dismutase and antimicrobial) have also been measured and compared with reported complexes.

Vet Rec, 2003 Oct 4, 153(14), 428 - 31
Antibiotic susceptibilities of recent isolates of Mycoplasma bovis in Belgium; Thomas A et al.; The susceptibilities of 40 recent Belgian field isolates of Mycoplasma bovis to 10 antimicrobial agents were assessed . Tiamulin was the most active antimicrobial agent against M bovis, with an initial inhibitory concentration (IIC50) of 0.06 microg/ml, but it is not licensed for the treatment of cattle . All three fluoroquinolones tested (danofloxacin, enrofloxacin and marbofloxacin) were effective against strains of M bovis, and had a minimum mycoplasmacidal concentration (MMC50) less than or equal to 1 microg/ml . Gentamicin was poorly effective, having an IIC50 of 8 microg/ml . Many strains of M bovis were resistant to tylosin, spectinomycin, lincomycin, tetracycline and oxytetracycline.

Crit Rev Microbiol, 2003, 29(3), 191 - 214
Antimicrobial properties of lysozyme in relation to foodborne vegetative bacteria; Masschalck B et al.; The purpose of this review is to describe the antibacterial properties and mode of action of lysozyme against gram-positive and gram-negative bacteria, and to provide insight in the underlying causes of bacterial resistance or sensitivity to lysozyme . Such insight improves our understanding of the role of this ubiquitous enzyme in antibacterial defense strategies in nature and provides a basis for the development and improvement of applications of this enzyme as an antibacterial agent . The bactericidal properties of lysozyme are primarily ascribed to its N-acetylmuramoylhydrolase enzymic activity, resulting in peptidoglycan hydrolysis and cell lysis . However, an increasing body of evidence supports the existence of a nonenzymic and/or nonlytic mode of action . Because gram-negative bacteria, including some major foodborne pathogens, are normally insensitive to lysozyme by virtue of their outer membrane that acts as a physical barrier preventing access of the enzyme, several strategies have been developed to extend the working spectrum of lysozyme to gram-negative bacteria . These include denaturation of lysozyme, modification of lysozyme by covalent attachment of polysaccharides, fatty acids and other compounds, attachment of C-terminal hydrophobic peptides to lysozyme by genetic modification, and the use of outer membrane permeabilizing agents such as EDTA or polycations or permeabilizing treatments such as high hydrostatic pressure treatment.

Curr Opin Chem Biol, 2003 Oct, 7(5), 563 - 9
Mechanisms of resistance to antibiotics; Wright GD; Microbial resistance to antibiotics is manifested by changes in antibiotic permeability, alteration of target molecules, enzymatic degradation of the antibiotics, and efflux of antimicrobials from the cytosol . Bacteria and other microorganisms use all of these mechanisms to evade the toxic effects of antibiotics . Recent research on the molecular aspects of these mechanisms, often informed by atomic resolution structures of proteins, enzymes and nucleic acids involved in these processes, has deepened our understanding of antibiotic action and resistance and, in several cases, spurred the development of strategies to overcome resistance in vitro and in vivo.

Paediatr Drugs, 2003, 5(11), 723 - 40
Neonatal sepsis: epidemiology and management; Baltimore RS; Neonatal sepsis is uncommon (2-4 per 1000 live births in developed countries), but the rate increases dramatically in premature newborns and those born to mothers with infections or prolonged rupture of the fetal membranes . While infections caused by organisms contracted from the mother at birth have decreased in the past two decades, there has been an increase in nosocomial infections . Today, most infants with sepsis have been hospitalized in neonatal intensive care units for weeks or months because of extreme prematurity, or because of a congenital malformation or surgical condition . Antimicrobial therapy is usually begun prior to the isolation of a pathogen and is based upon knowledge of the likely microbes in the particular clinical situation.The number of antimicrobial agents that can be safely used in neonates is relatively small, and dose administration usually needs to be adjusted based upon birthweight and post-gestational age . The decision whether to treat with antimicrobials should be made with consideration of the history, physical examination, and laboratory data . One should also consider the effects of the use of antimicrobials on the flora of the care unit . Bacterial resistance in the resident flora of the unit has become a major problem where there has been indiscriminate use of broad-spectrum agents.

Biochemistry, 2003 Nov 4, 42(43), 12503 - 10
NMR solution structure of viscotoxin C1 from Viscum album species Coloratum ohwi: toward a structure-function analysis of viscotoxins; Romagnoli S et al.; The high resolution three-dimensional structure of the newly discovered plant viscotoxin C1, from the Asiatic Viscum album ssp . Coloratum ohwi, has been determined in solution by (1)H NMR spectroscopy at pH 3.6 and 285 K . The viscotoxin C1-fold, consisting of a helix-turn-helix motif and a short stretch of an antiparralel beta-sheet is very similar to that found for the highly similar viscotoxins A2 and A3 and for other related thionins . Different functional properties of members of the thionin family are discussed here in light of the structural and electrostatic properties . Among the very homologous family of alpha- and beta-thionins, known for their antimicrobial activity, the viscotoxin subfamily differs from the other members because of its high toxicity against tumoral cells . Key residues for the modulation of viscotoxin cytotoxicity have been identified on the basis of sequence and structural alignment.

Int J Oral Maxillofac Implants, 2003 Sep-Oct, 18(5), 706 - 11
Antimicrobial efficacy of semiconductor laser irradiation on implant surfaces; Kreisler M et al.; PURPOSE: This study was conducted to investigate the antimicrobial effect of an 809-nm semiconductor laser on common dental implant surfaces . MATERIALS AND METHODS: Sandblasted and acid-etched (SA), plasma-sprayed (TPS), and hydroxyapatite-coated (HA) titanium disks were incubated with a suspension of S . sanguinis (ATCC 10556) and subsequently irradiated with a gallium-aluminum-arsenide (GaAlAs) laser using a 600-microm optical fiber with a power output of 0.5 to 2.5 W, corresponding to power densities of 176.9 to 884.6 W/cm2 . Bacterial reduction was calculated by counting colony-forming units on blood agar plates . Cell numbers were compared to untreated control samples and to samples treated with chlorhexidine digluconate (CHX) . Heat development during irradiation of the implants placed in bone blocks was visualized by means of shortwave thermography . RESULTS: In TPS and SA specimens, laser irradiation led to a significant bacterial reduction at all power settings . In an energy-dependent manner, the number of viable bacteria was reduced by 45.0% to 99.4% in TPS specimens and 57.6% to 99.9% in SA specimens . On HA-coated disks, a significant bacterial kill was achieved at 2.0 W (98.2%) and 2.5 W (99.3%) only (t test, P < .05) . For specimens treated with CHX, the bacterial counts were reduced by 99.99% in TPS and HA-coated samples and by 99.89% in SA samples . DISCUSSION: The results of the study indicate that the 809-nm semiconductor laser is capable of decontaminating implant surfaces . Surface characteristics determine the necessary power density to achieve a sufficient bactericidal effect . The bactericidal effect, however, was lower than that achieved by a 1-minute treatment with 0.2% CHX . The rapid heat generation during laser irradiation requires special consideration of thermal damage to adjacent tissues . CONCLUSION: No obvious advantage of semiconductor laser treatment over conventional methods of disinfection could be detected in vitro.

Biopolymers, 2003 Oct, 70(3), 424 - 34
The solution structure of frenatin 3, a neuronal nitric oxide synthase inhibitor from the giant tree frog, Litoria infrafrenata; Brinkworth CS et al.; The peptide frenatin 3 is a major component of the skin secretion of the Australian giant tree frog, Litoria infrafrenata . Frenatin 3 is 22 amino acids in length, and shows neither antimicrobial nor anticancer activity . It inhibits the production of nitric oxide by the enzyme neuronal nitric oxide synthase at a micromolar concentration by binding to its regulatory protein, Ca2+ calmodulin, a protein known to recognize and bind amphipathic alpha-helices . The solution structure of frenatin 3 has been investigated using NMR spectroscopy and restrained molecular dynamics calculations . In trifluoroethanol/water mixtures, the peptide forms an amphipathic alpha-helix over residues 1-14 while the C-terminal eight residues are more flexible and less structured . The flexible region may be responsible for the lack of antimicrobial activity . In water, frenatin 3 exhibits some alpha-helical character in its N-terminal region .

J Dent Res, 2003 Nov, 82(11), 877 - 82
Intracellular calcium in signaling human beta-defensin-2 expression in oral epithelial cells; Krisanaprakornkit S et al.; Expression of human beta-defensins is correlated with differentiation in the oral epithelium, consistent with their function as part of the epithelial antimicrobial barrier . Because calcium is a known regulator of epithelial differentiation, we tested the hypothesis that calcium concentration mediates beta-defensin expression . Gingival epithelial cells were cultured in medium containing low calcium concentration (0.03 mM), then either changed to high extracellular calcium concentrations or stimulated with thapsigargin to release intracellular calcium stores in the presence or absence of BAPTA-AM, a calcium chelator . Human beta-defensin-2 (hBD-2) mRNA expression was rapidly induced by thapsigargin, and more slowly induced by high extracellular calcium . Induction of hBD-2 peptide was confirmed by immunofluorescence . BAPTA-AM inhibited hBD-2 induction by both thapsigargin and calcium in a dose-dependent fashion . In addition, BAPTA-AM inhibited hBD-2 induction by a bacterial stimulant . Collectively, these findings demonstrate that intracellular calcium is a critical mediator of hBD-2 expression . Abbreviations used in this study are: BAPTA-AM, 1,2-bis(2-aminophenoxy)-ethane-N,N,N',N'-tetra-acetic acid tetrakis (acetoxymethyl ester); DMSO, dimethylsulfoxide; F . nucleatum, Fusobacterium nucleatum; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HBDs, human beta-defensins; HGECs, human gingival epithelial cells; MAP, mitogen-activated protein; and RT-PCR, reverse-transcriptase/polymerase chain-reaction.

Z Naturforsch {C}, 2003 Sep-Oct, 58(9-10), 746 - 51
Antimicrobial compounds of fungi vectored by Clusia spp . (Clusiaceae) pollinating bees; Bicalho B et al.; The production of antimicrobial compounds by fungi associated with Clusia spp . pollinating bees (Trigona sp., Trigonini) was investigated in order to approach natural mechanisms of microbial density control within nest environment . By using a bioassay-guided approach based on bioautography and minimal inhibitory concentration (MIC), known alpha,beta-dehydrocurvularin and curvularin were isolated from Curvularia eragrostidis (CCT 5634) and Curvularia pallescens (CCT 5654), and known cochlioquinone A and isocochlioquinone A were isolated from Drechslera dematioidea (CCT 5631).

Z Naturforsch {C}, 2003 Sep-Oct, 58(9-10), 649 - 54
Two new 24-isopropenyl-lanostanoids from Tillandsia brachycaulos; Cantillo-Ciau Z et al.; The leaves of Tillandsia brachycaulos afforded two novel tetracyclic triterpenoids identified as (24S)-24-isopropenyl-29-nor-5alpha-lanosta-7-en-3beta-ol (1) and (24S)-24-isopropenyl-29-nor-5alpha-lanosta-7-en-3-one (2), in addition to the known isopimaric acid (3) and chlorogenic acid (4) . 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 antimicrobial and antifungal activities were studied . The compounds did not show significant activity.

Kekkaku, 2003 Sep, 78(9), 601 - 4
{Role of new quinolones in the treatment of mycobacteriosis}; Controlled expression of an rpoS antisense RNA can inhibit RpoS function in Escherichia coli; Department of Biology, McMaster University, Hamilton, Ontario L8S 4K1, CanadaWe show that an inducible rpoS antisense RNA complementary to the rpoS message can inhibit expression of RpoS in both exponential and stationary phases and can attenuate expression of the rpoS regulon in Escherichia coli . Plasmids containing rpoS antisense DNA expressed under the control of the T7lac promoter and T7 RNA polymerase were constructed, and expression of the rpoS antisense RNA was optimized in the pET expression system . rpoS antisense RNA levels could be manipulated to effectively control the expression of RpoS and RpoS-dependent genes . RpoS expression was inhibited by the expression of rpoS antisense RNA in both exponential and stationary phases in E . coli . RpoS-dependent catalase HPII was also downregulated, as determined by catalase activity assays and with native polyacrylamide gels stained for catalase . Induced RpoS antisense expression also reduced the level of RpoS-dependent glycogen synthesis . These results demonstrate that controlled expression of antisense RNA can be used to attenuate expression of a regulator required for the expression of host adaptation functions and may offer a basis for designing effective antimicrobial agents.

Antimicrob Agents Chemother, 2003 Nov, 47(11), 3435 - 41
Synthesis, degradation, and antimicrobial properties of targeted macromolecular prodrugs of norfloxacin; Roseeuw E et al.; Long-term antibiotic treatment is required to cure tuberculosis . Targeted antibiotics should improve the efficacy of treatment by concentrating the drugs close to the bacteria . The aim of the present study was to synthesize targeted conjugates . For this purpose, we used mannose as a homing device to direct norfloxacin into macrophages . Dextran was used as the polymer bearing both mannose and norfloxacin . Using different peptide spacer arms to link norfloxacin to dextran, we demonstrated that norfloxacin acts as an antibiotic only when it is released in its native form . Also, targeting by using mannose as a homing device is required to achieve antimycobacterial activity in vivo . Thus, norfloxacin, which is inactive against mycobacteria in its native form in vivo, can be transformed into an active drug by targeting.

Biochem Biophys Res Commun, 2003 Nov 7, 311(1), 90 - 7
Antimicrobial peptides from scorpion venom induce Ca(2+) signaling in HL-60 cells; Moerman L et al.; Parabutoporin (PP) and opistoporin 1 (OP1) are amphipathic alpha-helical antimicrobial peptides that were recently isolated from scorpion venom . In assays in which single granulocyte-like HL-60 cells as well as cells in suspension were used, both peptides were able to induce a reversible Ca(2+) release from intracellular stores and to increase Ca(2+) influx . Both effects could be clearly differentiated for OP1, inducing Ca(2+) release at lower concentrations . The Ca(2+) release was pertussis toxin-sensitive indicating the involvement of G-proteins . Ca(2+) release depended on the stage of differentiation of the cells with undifferentiated cells being the most sensitive . Desensitization occurred with OP1 . No cross-desensitization occurred between OP1 and the bacterial chemoattractant fMLP indicating the involvement of different types of receptors . Ca(2+) release by OP1 was found not to be mediated via interaction with the formyl peptide receptor-like 1 . Although some of the results might favor a receptor-like interaction, the receptor involved could not be identified.

Kansenshogaku Zasshi, 2003 Sep, 77(9), 654 - 60
{Detection of MRSA with antimicrobial susceptibility by using chemiluminescent assay}; Takakura R et al.; Chemiluminescent assay can give the result of the detection of MRSA about 13 hours more rapidly than conventional broth microdilution method . In order to apply chemiluminescent assay to detection of MRSA, we compared MIC and antimicrobial susceptibility to MPIPC in using chemiluminescent assay with these in using broth microdilution method . In MSSA, rate of concordance of MIC and antimicrobial susceptibility to MPIPC obtained by both methods was 87%, but all MICs come to be agreed by modifying the concentration of bacterial liquid . In MRSA, all MICs and susceptibility to MPIPC are agreed in both methods . Although we have used chemiluminescent assay to detect MRSA for one year, no trouble has been reported by clinical side . The chemiluminescent assay is evaluated to be good in detecting MRSA.

Zhonghua Nan Ke Xue, 2003 Sep, 9(6), 403 - 6
{Some controversial conditions in the management of chronic prostatitis/chronic pelvic pain syndrome}; Tang XD; Chronic prostatitis (CP)/chronic pelvic pain syndrome (CPPS) is a common problem of medically controversial condition that causes considerable morbidity and impact on life . Although there are many competing causes proposed, the etiology and pathogenesis of CP/CPPS remain unclear . The causative factors underlying the CPPS are not fully understood . The optimal management of CP/CPPS is still unknown . The guideline of diagnosis and management of CP/CPPS based on evidence base medicine is not yet established . Many problems are still not resolved, such as the significance of leukocytes and the role of inflammation in CP/CPPS, the significance of bacteria presence and the role of infection in CP/CPPS, the correlation between leukocytes/bacteria and severity of symptoms, how to divide the subgroups of CP/CPPS, the role of antimicrobial therapy in the treatment of men with CP/CPPS, why patients with category IIIb complain of symptoms, while those with category IV complain of none . Although CP/CPPS is now achieving greater recognition, well-designed studies with large sample size should be performed.

Infect Immun, 2003 Nov, 71(11), 6148 - 54
Characterization of chlamydial genital infection resulting from sexual transmission from male to female guinea pigs and determination of infectious dose; Rank RG et al.; A major problem in the study of chlamydial genital infections in animal models has been the use of varied doses of chlamydiae for infection in different laboratories . It is clearly desirable to use a dose which approximates that of natural sexual infection, but that dose to date has not been determined because of the inability of researchers to quantify chlamydiae in semen . Fortunately, sexual transmission of chlamydiae has been described for the guinea pig model of infection with the chlamydial agent of guinea pig inclusion conjunctivitis (GPIC) . In this study, we undertook to determine the approximate infection dose in actual sexual transmission by comparing the kinetics of infection in female guinea pigs acquired via sexual contact to those of genital infections induced artificially with known quantities of chlamydiae . Groups of guinea pigs were infected intravaginally with 10(4), 10(3), 10(2), and 10(1) inclusion-forming units (IFU) of GPIC, and the kinetics of the infection were determined . Infection with 10(2) IFU produced infections with lower peak levels than those in animals receiving 10(4) or 10(3) IFU . Seventy percent of animals receiving 10(2) IFU became infected, while 100 and 79% of animals receiving 10(4) and 10(3) IFU, respectively, became infected . Animals receiving 10(2) IFU also had a longer incubation period . Of 19 animals that mated with infected males, 63.2% became infected, with an infection course which was not significantly different than that of the 10(2)-IFU-infected group . The data suggest that female guinea pigs received approximately 10(2) IFU by sexual transmission . Of interest was the observation that the guinea pigs infected by sexual transmission shed organisms for a significantly shorter time period than that of any group that was artificially infected . This result suggests that there may be factors associated with semen which passively transfer antimicrobial activity to the female or enhance the innate host response in the female . Immunization of females with an inactivated vaccine was also found to elicit a protective immune response against sexual challenge, demonstrating that the model can be used in the evaluation of possible vaccine candidates and/or methodologies . There is currently no other animal model available for any sexually transmitted disease in which the disease or the ability to prevent the disease may be studied in animals infected by the natural means.

Farmaco, 2003 Nov, 58(11), 1105 - 11
Substituted 5-aroylpyrazine-2-carboxylic acid derivatives: synthesis and biological activity; Dolezal M et al.; Homolytic aroylation of pyrazine nucleus with various substituted aromatic carbaldehydes afforded a series of 5-aroylpyrazine-2-carboxylic acid derivatives . The synthetic approach, analytical and spectroscopic data of all compounds synthesized, their preliminary in vitro evaluation of antituberculotic and antifungal activities, cytotoxicity data and subsequent SAR studies are presented . Among all derivatives prepared, only 5-(4-chlorobenzoyl)-pyrazine-2-carbothioamide (3d) showed promising activity (90% inhibition) against Mycobacterium tuberculosis . The highest antifungal effect (MIC<1.95 microM ml(-1)) against Trichophyton mentagrophytes, the most susceptible fungal strain tested, was found for 5-benzoylpyrazine-2-carbothioamide (3a) . Thioamides exhibited higher in vitro antimicrobial activity than the corresponding amides.

Enferm Infecc Microbiol Clin, 2003 Nov, 21(9), 520 - 8; quiz 529, 533
{Tetracyclines, sulfonamides and metronidazole}; Perez-Trallero E et al.; Tetracyclines form a group of natural and semisynthetic products that acts inhibiting the bacterial protein synthesis . They are bacteriostatic agents, exhibiting activity against a wide range of organisms, but they are at the present of limited use because of their acquired resistance . Doxycycline is currently the most frequently used tetracycline in human medicine and it is included in the List of Essential Medicines of the World Health Organization . Sulfonamides are synthetic, broad-spectrum bacteriostatic antibiotics . They were the first effective systemic antimicrobial agents . Their mode of action is based on the inhibition of DNA synthesis . Due to their toxicity and high adquired resistance their use is currently very low . Metronidazole is the main compound of 5-nitroimidazole family . It is a very active bactericidal antibiotic against anaerobic and some microaerophilic bacteria and it is still very useful in the treatment of bacterian and parasitic infections.

J Food Prot, 2003 Oct, 66(10), 1783 - 9
Enhanced inhibition of Escherichia coli O157:H7 by lysozyme and chelators; Boland JS et al.; This study examined the effects of three chelating agents (EDTA, disodium pyrophosphate {DSPP}, and pentasodium tripolyphosphate {PSTPP}) on the inhibition of the growth of Escherichia coli O157:H7 by lysozyme . The objective of this study was to identify replacement chelators that exhibit synergistic properties similar to those of EDTA . The inhibitory effects of EDTA at 300 to 1,500 microg/ml and of DSPP and PSTPP at 3,000 to 15,000 microg/ml in combination with lysozyme at 200 to 600 microg/ml for up to 48 h at pHs of 6.0, 7.0, and 8.0 on four strains of E . coli O157:H7 was studied with the use of a microbroth dilution assay . The addition of EDTA enhanced lysozyme's inhibitory effect on strains of E . coli O157:H7 . EDTA at > or = 300 microg/ml combined with lysozyme at 200 to 600 microg/ml was sufficient to inhibit the growth of the strains at pHs of 6.0 and 8.0 . At pH 7.0, lysozyme at 200 to 600 microg/ml and EDTA concentrations of > or = 1,000 microg/ml were effective in inhibiting three of the four strains . DSPP at pH 6.0 was inhibitory at > or = 10,000 microg/ml when combined with lysozyme at 200 to 300 microg/ml . In contrast, PSTPP increased the inhibitory activity of lysozyme more effectively at pH 8.0 . Lysozyme at 200 to 600 microg/ml was effective against two strains of E . coli O157:H7 when used in conjunction with PSTPP at > or = 5,000 microg/ml . The remaining strains were inhibited by PSTPP at > or = 10,000 microg/ml . Our results indicate that inhibition occurred with each lysozyme-chelator combination, but the concentrations of phosphates required to increase the antimicrobial spectrum of lysozyme against E . coli O157:H7 were higher than the EDTA concentrations required to achieve the same effect.

Gut, 2003 Nov, 52(11), 1591 - 7
Expression of NOD2 in Paneth cells: a possible link to Crohn's ileitis; Ogura Y et al.; BACKGROUND AND AIMS: Genetic variation in NOD2 has been associated with susceptibility to Crohn's disease (CD) and specifically with ileal involvement . The reason for the unique association of NOD2 mutations with ileal disease is unclear . To identify a possible link, we tested expression of NOD2 in intestinal tissue of CD patients and controls . PATIENTS AND METHODS: Fifty five specimens of ileum or colon from 21 CD patients, seven ulcerative colitis (UC) patients, and five controls with pathology other than CD or UC were stained for NOD2 using an immunoperoxidase method . RESULTS: Using a monoclonal antibody against NOD2 developed in our laboratory, we detected uniform expression of NOD2 in terminal ileum Paneth cells from controls and patients as well as in metaplastic Paneth cells in the colon . Mechanical purification showed enriched expression of NOD2 mRNA in ileal crypts . In Paneth cells, NOD2 was located in the cytosol in close proximity to the granules that contain antimicrobial peptides . We detected minimal NOD2 in the villous epithelium of the ileum or in the colonic epithelium from both CD patients and controls . CONCLUSIONS: These results suggest a role for NOD2 in the regulation of Paneth cell mediated responses against intestinal bacteria and a plausible mechanism to explain the selective association of NOD2 mutations with ileal disease . The impaired capacity of CD associated mutations to sense luminal bacteria may result in increased susceptibility to certain gut microbes.

Curr Microbiol, 2003 Sep, 47(3), 244 - 9
A combination effect of epigallocatechin gallate, a major compound of green tea catechins, with antibiotics on Helicobacter pylori growth in vitro; Yanagawa Y et al.; Since green tea catechins are known to have antimicrobial activity against a variety of microorganisms, their possible effects on Helicobacter pylori in combination with antibiotics were examined . Fifty-six clinical isolates of H . pylori, including 19 isolates highly resistant to metronidazole (MTZ) and/or clarithromycin (CLR), were used to determine in vitro sensitivity to tea catechins . The MIC90 of both epigallocatechin gallate (EGCg) and epicatechin gallate (ECg) was 100 microg/ml . However, other tea catechins tested did not show any anti-H . pylori activity . Highly antibiotic-resistant clinical isolates showed a similar sensitivity to both EGCg and ECg . The kinetic study of antibacterial activity in liquid cultures revealed a relatively slow but strong activity on the growth of H . pylori . In combination with sub-MIC of amoxicillin (AMX), the antibacterial activity of AMX was significantly enhanced by the presence of EGCg . To estimate the general combination effect between EGCg and other antibiotics, such as MTZ and CLR, on the antibacterial activity against clinical isolates, the fraction inhibitory concentration (FIC) was determined by checkerboard study . The FIC indexes showed additive effects between EGCg and antibiotics tested . These results indicatethat EGCg may be a valuable therapeutic agent against H . pylori infection.

Exp Appl Acarol, 2002, 28(1-4), 135 - 40
Involvement of antibacterial peptide defensin in tick midgut defense; Nakajima Y et al.; Animals are constantly threatened by pathogenic microorganisms and have developed cellular and humoral immune responses to combat these infections . Invertebrates possess only an innate non-specific immune response . Antimicrobial substances are major components of innate immunity not only in invertebrates but also in vertebrates . Despite the importance of ticks as vectors of disease very little is known about their immune system . Our recent studies have revealed that four defensin isoforms are present in Ornithodoros moubata . These four isoforms are constitutively expressed in the midgut and up-regulated in response to blood feeding . Moreover, a mature peptide of defensin isoform A has been isolated from the tick midgut lumen . These findings indicate Ornithodoros defensins are involved in tick midgut defense against potentially harmful invasive microbes.

Biochem Cell Biol, 2003 Oct, 81(5), 349 - 54
Expression of bovine lactoferrin and lactoferrin N-lobe by recombinant baculovirus and its antimicrobial activity against Prototheca zopfii; Tanaka T et al.; Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in secretory fluids of mammals . In this study, DNA encoding bovine lactoferrin (bLF) or the N-terminal half of bLF (bLF N-lobe) was inserted into a baculovirus transfer vector, and a recombinant virus expressing bLF or bLF N-lobe was isolated . An 80-kDa bLF-related protein expressed by the recombinant baculovirus was detected by monoclonal antibodies against bLF N-lobe and the C-terminal half of bLF (bLF C-lobe) . A 43-kDa bLF N-lobe-related protein expressed by the recombinant baculovirus was detected by anti-bLF N-lobe monoclonal antibody, but not by anti-bLF C-lobe monoclonal antibody . These proteins were also secreted into the supernatant of insect cell cultures . Recombinant bLF (rbLF) and bLF N-lobe (rbLF N-lobe) were affected by tunicamycin treatment, indicating that rbLF and rbLF N-lobe contain an N-linked glycosylation site . Antimicrobial activity of these recombinant proteins against Prototheca zopfii (a yeast-like fungus that causes bovine mastitis) was evaluated by measuring the optical density of the culture microplate . Prototheca zopfii was sensitive to rbLF and rbLF N-lobe, as well as native bLF . There was no difference in antimicrobial activity between rbLF N-lobe and bLF C-lobe.

Mol Immunol, 2003 Nov, 40(7), 469 - 75
Intestinal defensin gene expression in human populations; Dhaliwal W et al.; Defensins are thought to play a major role in the defence of small intestinal crypts against colonisation by potential pathogens . In humans two alpha-defensins, HD5 and HD6 and two beta-defensins, hBD1 and hBD2, probably contribute to the antimicrobial barrier, but there are no data to indicate how the expression of these defensin genes might vary in individuals and in populations . To begin to address this question we developed a competitive reverse transcriptase polymerase chain reaction (RT-PCR) assay to quantify HD5 and HD6 mRNA and used it to measure transcripts in small intestinal biopsy tissue from adults living in London, UK, or in Lusaka, Zambia . We also measured alpha- and beta-defensin mRNA in biopsies collected in London from different regions of the small intestine . Jejunal biopsies (n=169) from 83 adults in Lusaka contained approximately one order of magnitude less HD5 and HD6 mRNA than biopsies (n=33) obtained from 27 adults in London . HD5 and HD6 transcript levels were high throughout duodenum, jejunum and ileum . hBD1 and hBD2 mRNA were detected in some, but not all, biopsies from normal small intestine . These data suggest that alpha-defensin expression is down-regulated in tropical populations, and that there are distinct pathways regulating transcription of alpha- and beta-defensins.

Mol Immunol, 2003 Nov, 40(7), 457 - 62
Defensins and acne; Philpott MP; Acne a disease of the pilosebaceous unit is characterised by hypercornification and hyperkeratosis of outer root sheath (ORS) and sebaceous duct and perilesional infiltrate . Lesions may be characterised as "non"-inflammatory versus inflammatory . Hypercornification of the distal ORS and the pilosebaceous duct in concert with increased sebum production and abnormalities of the microbial flora are considered to be major factors in the pathogenesis of acne vulgaris . However, the basic mechanisms involved in the development of inflammation during acne vulgaris remain unclear . We have investigated the expression patterns of two antimicrobial peptides, human beta-defensin 1 (hBD1) and human beta-defensin 2 (hBD2) in healthy human hair follicles as well as in peri- and intralesional skin of acne vulgaris lesions such as comedones, papules and pustules . Strong hBD1 and hBD2 immunoreactivity was found in all suprabasal layers of the epidermis, and all permanent compartments of the hair follicle including the distal ORS of the hair follicle and the pilosebaceous duct . Moreover, marked hBD1 and hBD2 expression was also detected in the hair follicle stem cell compartment . In contrast, the proximal follicle bulb which undergoes apoptotic regression and is also able to regenerate following injury did not express hBD1 or hBD2 . The majority of acne biopsies displayed a marked upregulation of hBD2 IR in the lesional and perilesional epithelium; in particular in pustules, and a less marked upregulation of hBD1 IR . The upregulation of beta-defensins expression in acne vulgaris lesions when compared to controls suggests that beta-defensins may be involved in the pathogenesis of acne vulgaris.

Mol Immunol, 2003 Nov, 40(7), 445 - 50
Regulation of expression of beta-defensins: endogenous enteric peptide antibiotics; O'Neil DA; Evidence for the central role that intestinal beta-defensins play in maintaining gut health continues to accumulate within the literature . Two epithelially-derived enteric beta-defensins, hBD1 and hBD2, have been identified thus far and the following chapter reviews our current understanding of how the expression and secretion of these endogenous antimicrobial, chemotactic and adjuvant peptides is regulated within the context of the most microbe-rich of mucosal environments, the gastrointestinal tract . The agonists and microbial moieties identified as being responsible for the direct receptor-mediated induction of enteric epithelial beta-defensins, the signaling and nuclear events that are triggered as a consequence and which drive defensin gene transcription, the potential antimicrobial and immunomodulatory consequences of beta-defensin release within the luminal and mucosal aspects of the alimentary tract thereafter and the validity of animal models for the study of these key immune effector molecules in vivo are discussed . These significant and recent discoveries have provided much in the way of momentum for the pace with which this exciting and dynamic area of mucosal immunology research continues to move forward.

Mol Immunol, 2003 Nov, 40(7), 413 - 21
Rapid sequence divergence in mammalian beta-defensins by adaptive evolution; Maxwell AI et al.; beta-Defensin genes encode broad spectrum antimicrobial cationic peptides . We have analysed the largest murine and human clusters of these genes, which localise to mouse and human chromosome 8 . Using hidden Markov models, we identified novel mouse and human beta-defensin genes . We subsequently found full-length expressed transcripts for these novel genes . Expression in the mouse was high in brain and reproductive tissues . Fourteen murine beta-defensins could be grouped into two clear sub-groups by virtue of their position and high signal sequence (exon 1 encoded) identity . In contrast, there was a very low level of sequence conservation in the exon 2 region encoding the mature antimicrobial peptide . Evolutionary analysis revealed strong evidence that following gene duplication, exon 1 and surrounding non-coding DNA show little divergence within subfamilies . The focus for rapid sequence divergence is localised in the DNA encoding the mature peptide and this is driven by accelerated positive selection . In the human we also conclude that the locus has evolved by successive rounds of duplication followed by substantial divergence involving positive selection, to produce a diverse cluster of paralogous genes prior to human-baboon divergence . This mechanism of adaptive evolution is consistent with the role of this gene family as defence against bacterial pathogens . In order to look at function of these rapidly evolving genes, we characterised one of the novel mouse beta-defensin genes . This gene deviates from the canonical six cysteine motif present in the mature functional peptide of all other beta defensins . This defensin related gene (Defr1) is most highly expressed in testis and heart and the genomic organisation is highly similar to Defb3-6 . A synthetic Defr1 peptide was shown to exist as a dimer and yet displayed both antimicrobial and chemotactic activity . The antimicrobial activity of Defr1 against S . aureus, E . coli and B . cepacia was found to be reduced in raised concentration of NaCl, but its action against P . aeruginosa was independent of NaCl concentration . These data have major implications on the structure and functions of these important host defence molecules.

Mol Immunol, 2003 Nov, 40(7), 395 - 405
Lactoferrin--a multifunctional protein with antimicrobial properties; Farnaud S et al.; Lactoferrin is a member of the transferrin family of iron-binding proteins . Numerous functions have been reported and continue to be reported for the protein, some of which are related to its iron-binding properties . Its extensive antimicrobial activities were originally attributed to its ability to sequester essential iron, however, it is now established that it possesses bactericidal activities as a result of a direct interaction between the protein or lactoferrin-derived peptides . This article reviews the antimicrobial activities of lactoferrin and discusses the potential mode of action of lactoferrin-derived cationic peptides against Gram-negative bacteria in the light of recent studies.

Clin Chim Acta, 2003 Nov, 337(1-2), 11 - 21
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in clinical chemistry; Marvin LF et al.; Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-Tof-MS) has recently become a popular and versatile method to analyze macromolecules from biological origin . In this paper, we will review the application of MALDI-Tof-MS in clinical chemistry and biology . MALDI-Tof-MS is used in clinical chemistry, e.g . disease markers can be identified with MALDI-MS analysis in combination with 1-D and 2-D gel electrophoresis separations thanks to either peptide mass fingerprinting (PMF) or peptide sequence tag (PST) followed by data base searching . In microbiology, MALDI-Tof-MS is employed to analyze specific peptides or proteins directly desorbed from intact viruses, bacteria and spores . The capability to register biomarker ions in a broad m/z range, which are unique and representative for individual microorganisms, forms the basis of taxonomic identification of bacteria by MALDI-Tof-MS . Moreover, this technique can be applied to study either the resistance of bacteria to antibiotics or the antimicrobial compounds secreted by other bacterial species . More recently, the method was also successfully applied to DNA sequencing (genotyping) as well as screening for mutations . High-throughput genotyping of single-nucleotide polymorphisms has the potential to become a routine method for both laboratory and clinical applications . Moreover, posttranscriptional modifications of RNA can be analyzed by MALDI using nucleotide-specific RNAses combined with further fragmentation by post source decay (PSD).

FEMS Microbiol Lett, 2003 Oct 10, 227(1), 107 - 11
Testing the nematophagous biological control strain Paecilomyces lilacinus 251 for paecilotoxin production; Khan A et al.; Paecilomyces lilacinus is a nematophagous fungus currently developed as a biological control agent . In order to evaluate potential toxin production, culture extract and concentrated culture supernatant of P . lilacinus strain 251 were tested against Gram-negative and Gram-positive bacteria . High-performance liquid chromatography analysis was carried out to compare the chromatograms of P . lilacinus strain 251 with the chromatogram of known paecilotoxin . It was found that the 251 strain of P . lilacinus did not produce detectable levels of paecilotoxin or other toxins with antimicrobial activity.

Phytochemistry, 2003 Nov, 64(6), 1061 - 7
Hofmannolin, a cyanopeptolin from Scytonema hofmanni PCC 7110; Matern U et al.; Two depsipeptide metabolites, scyptolin A and B, were reported recently from the axenically grown terrestrial cyanobacterium Scytonema hofmanni PCC 7110 . A related, novel depsipeptide was isolated from this Scytonema and designated hofmannolin . The amino acid analysis in context with infrared, mass and 1H/13C-NMR spectroscopies revealed a cyclic depsipeptide structure of M(r) 1073 belonging to the class of cyanopeptolins . Two peculiar features distinguish hofmannolin from other cyanopeptolins: O-methylated tyrosine forms the sixth moiety from the amino terminus, and the N-terminus is blocked by 2-hydroxy-3-methyl-valeric acid, a residue that has not yet been reported as a component in other cyanopeptolins . Preliminary assays of peptidase inhibitory and antimicrobial activities suggested negligible bioactivities for hofmannolin.

Biochemistry, 2003 Oct 28, 42(42), 12251 - 9
Interaction of antimicrobial peptides with lipopolysaccharides; Ding L et al.; We study the interaction of antimicrobial peptides with lipopolysaccharide (LPS) bilayers to understand how antimicrobial peptides interact with the LPS monolayer on the outer membrane of Gram-negative bacteria . LPS in water spontaneously forms a multilamellar structure composed of symmetric bilayers . We performed X-ray lamellar diffraction and wide-angle in-plane scattering to study the physical characteristics of LPS multilayers . The multilayer alignment of LPS is comparable to phospholipids . Thus, it is suitable for the application of oriented circular dichroism (OCD) to study the state of peptides in LPS bilayers . At high hydration levels, the chain melting temperature in multilamella detected by X-ray diffraction is the same as that of LPS aqueous dispersions, as measured by calorimetry . LPS has a strong CD, but with a careful subtraction of the lipid background, the OCD of peptides in LPS is measurable . The method was tested successfully with melittin . It was then applied to two representative antimicrobial peptides, magainin and protegrin . At peptide concentrations comparable to the physiological conditions, both peptides penetrate transmembrane in LPS bilayers . The results imply that antimicrobial peptides readily penetrate the LPS monolayer of the outer membrane.

Neth J Med, 2003 Jul, 61(7), 233 - 4
Immunomodulation by antimicrobial drugs; van der Meer JW; Immunomodulation aims at either enforcement of host defence mechanisms or dampening of the inflammatory response of the host . Thus, immunomodulatory drugs may enhance host defence by either stimulating the inflammatory response or inhibiting the counter-regulatory, anti-inflammatory response . On the other hand, the response of the host may be down-modulated through inhibition of the inflammatory response or induction of counter-regulatory, anti-inflammatory mechanisms . Antimicrobial drugs may have such immunomodulatory effects, but so far these effects have not been exploited clinically.

J Am Vet Med Assoc, 1999 Dec 1, 215(11), 1671 - 4
Usefulness of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis in animals; Massa KL et al.; OBJECTIVE: To determine the diagnostic value of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis (IUK) . DESIGN: Prospective study . ANIMALS: 48 animals (26 dogs, 13 horses, 7 cats, 1 bird, and 1 llama) with corneal ulcers . PROCEDURE: Scrapings from corneal ulcers were examined cytologically . Corneal swab specimens were submitted for microbial culture . Animals were grouped according to whether they had been receiving antimicrobials at the time of admission . RESULTS: Of the 38 animals receiving antimicrobials, 19 had positive results for IUK on cytologic evaluation, 20 on microbial culture, and 26 on cytologic evaluation, microbial culture, or both . Of the 10 animals not receiving antimicrobials at the time of admission, 7 had positive results for IUK on cytologic evaluation, and 9 had positive results on microbial culture . In this group of 10 animals, additional animals with IUK were not identified on the basis of cytologic evaluation alone . When all 48 animals were considered irrespective of antimicrobial treatment, 26 and 29 had positive results for IUK on cytologic evaluation and microbial culture, respectively, whereas IUK was confirmed in 35 animals on the basis of cytologic evaluation, microbial culture results, or both . CONCLUSIONS AND CLINICAL RELEVANCE: Microbial culture and cytologic evaluation of corneal specimens maximizes identification of IUK, especially in animals receiving antimicrobial treatment . Because of serious consequences of untreated IUK, we recommend that both diagnostic tests be used to tailor treatment and reduce risk of vision impairment in animals.

J Foot Ankle Surg, 2003 Sep-Oct, 42(5), 302 - 4
What is the shelf life of physician-mixed antibiotic-impregnated calcium sulfate pellets?
Armstrong DG, Stephan KT, Espensen EH, Lipsky BA, Boulton AJ.
This pilot study was undertaken to evaluate the short-term in vitro antimicrobial stability of both vancomycin- and tobramycin-impregnated calcium sulfate pellets mixed and stored in a clinical setting . Powdered tobramycin sulfate (500 mg) and vancomycin hydrochloride (500 mg) were blended into separate basins containing 25 g of surgical-grade calcium sulfate powder, then mixed with 8 mL of sterile saline . From this admixture, 6.0-mm pellets were produced . These were removed from the sterile container (stored at room temperature) at 1, 7, 30, 60, 90, and 120 days and tested against a variety of pathogenic bacterial isolates by using a modification of the standardized Kirby-Bauer test . Control pellets containing no antibiotic were also evaluated . There was no inhibition of bacterial growth by the non-antibiotic-impregnated (control) pellets . There was no appreciable difference in the zones of inhibition for any of the organisms with pellets stored for 1, 7, 30, 60, 90, or 120 days . Zones of inhibition for the various antibiotics to the strain of organism tested ranged from 17 mm to 30 mm, depending on the pathogenic isolate and the antibiotic evaluated . The zones of inhibition observed were similar to those designating antibiotic susceptibility in the Kirby-Bauer test . The results of this preliminary study suggest that clinician-mixed calcium sulfate pellets containing either vancomycin or tobramycin, when stored under normal room temperature and ambient humidity, appear to maintain their antimicrobial characteristics for at least 120 days.

Med Pregl, 2003 May-Jun, 56(5-6), 243 - 6
{Aging and infection}; Vukadinov J et al.; EPIDEMIOLOGY: Aging is a natural process and a part of our lives, but nowadays there is an increase in the number of persons aged 65 and over . Today infectious diseases are still responsible for one-third of all deaths in the world . The elderly population is most vulnerable to serious infections and at greatest risk for death and complications . Among geriatric population pneumonia and influenza are the fourth most common cause of death . VACCINATION: One of the goals of preventive medicine is to reduce the rate of complications and mortality from infectious diseases by increasing immunization rates . Influenza and pneumococcal vaccines are indicated for persons aged 65 and over . Despite well-recognized benefit of such vaccination, less than 50% of eligible patients receive the vaccine each year . INFECTIONS: Older persons generally have increased susceptibility to infections because of multiple risk factors and they are the most vulnerable population to nosocomial and health-care associated infections . Older persons may manifest infectious diseases atypically, with acute confusion or delirium which can lead into delay in diagnosis and therapy . It is important to know that the older present with delayed or poor response to antimicrobial therapy and high rates of adverse reactions to drugs, including antibiotics . CONCLUSION: As elderly population is rapidly growing, majority of patients with serious or life-threatening infections are old . Geriatric issues have not typically been a focus of training in infectious diseases, but we must become aware of and knowledgeable about special and unique aspects of infections in this population.

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 2003 Sep, 20(3), 451 - 4
{Airway beta-defensin-2 gene transfer enhances the bacterial clearing of rat lung}; Zhou H et al.; beta-defensins possess a broad spectrum of antimicrobial activity . In this study its in vivo antibacterial effect was evaluated by using gene transfer . Rat beta-defensin-2 (rBD2) recombinant pBK-CMV-rBD2 and pCD-NA-3, 1-Myc-His(+)-rBD2 were constructed . Then, by use of liposome agent, the recombinants were delivered into rat airway via tracheal injection . The rBD-2 mRNA expression was detected in the trachea by RT-PCR and its protein expression was determined in the lungs by the tag His immunostaining, 24 hours after inoculation via trachea, the count of P . areuginosa in the lung of rat transfected with pBK-CMV-rBD2 markedly decreased, compared with the control (n = 8, P = 0.003) . The data presented in this study provide evidence that airway beta-defensin-2-gene transfer can protect the rat against bacterial infection in vivo, suggesting the beta-defensins as part of the innate host defense system can be of potential applicability.

J Antimicrob Chemother, 2003 Nov, 52(5), 764 - 71 Epub 2003 Oct 16.
Room for improvement: a systematic review of the quality of evaluations of interventions to improve hospital antibiotic prescribing; Ramsay C et al.; INTRODUCTION: In 1999, the British Society for Antimicrobial Chemotherapy (BSAC) and Hospital Infection Society (HIS) convened a working party on optimization of antibiotic prescribing in hospitals . This study was undertaken in order to evaluate the current evidence base on the effectiveness of interventions to change antibiotic prescribing to hospital inpatients . METHODS: We have systematically reviewed the literature from 1980 to identify interventions that alone, or in combination, are effective in improving antibiotic prescribing to hospital inpatients . The protocol was peer reviewed and has been published by the Effective Practice and Organization of Care (EPOC) Group of the Cochrane Collaboration . RESULTS: We identified 306 papers, of which 91 (30%) met the minimum inclusion criteria for a Cochrane EPOC review . The reasons for exclusion were uncontrolled before and after design (141/306; 46%) and inadequate interrupted time series (74/306; 24%) with fewer than three observations before and after the intervention . Most of the rejected interrupted time series (ITS) studies had only one or two data points before the intervention with many (up to 15) after it . Only 15 (40%) of the 38 included ITS studies had a statistical analysis and 11 of these used inappropriate statistical tests (e.g . t-test of pre- and post-intervention mean data) rather than analysis of time trends . Regression analysis of the proportion of included studies by year of publication did show a significant positive correlation (R2 = 0.7886) . Nonetheless, of 47 papers published since 2000, only 19 (40%) met the minimum eligibility criteria . CONCLUSIONS: The majority of evaluations used fundamentally flawed methodology . There is limited evidence of improvement over time . These problems could be resolved if researchers and referees of protocols or manuscripts implemented the EPOC methodology.

Am J Respir Crit Care Med, 2004 Jan 15, 169(2), 187 - 94 Epub 2003 Oct 16.
Cathelicidin peptide sheep myeloid antimicrobial peptide-29 prevents endotoxin-induced mortality in rat models of septic shock; Giacometti A et al.; The present study was designed to investigate the antiendotoxin activity and therapeutic efficacy of sheep myeloid antimicrobial peptide (SMAP)-29, a cathelicidin-derived peptide . The in vitro ability of SMAP-29 to bind LPS from Escherichia coli 0111:B4 was determined using a sensitive limulus chromogenic assay . Two rat models of septic shock were performed: (1) rats were injected intraperitoneally with 1 mg E . coli 0111:B4 LPS and (2) intraabdominal sepsis was induced via cecal ligation and single puncture . All animals were randomized to receive parenterally isotonic sodium chloride solution, 1 mg/kg SMAP-29, 1 mg/kg polymyxin B or 20 mg/kg imipenem . The main outcome measures were: abdominal exudate and plasma bacterial growth, plasma endotoxin and tumor necrosis factor-alpha concentrations, and lethality . The in vitro study showed that SMAP-29 completely inhibited the LPS procoagulant activity at approximately 10 microM peptide concentration . The in vivo experiments showed that all compounds reduced the lethality when compared with control animals . SMAP-29 achieved a substantial decrease in endotoxin and tumor necrosis factor-alpha plasma concentrations when compared with imipenem and saline treatment and exhibited a slightly lower antimicrobial activity than imipenem . No statistically significant differences were noted between SMAP-29 and polymyxin B . SMAP-29, because of its double antiendotoxin and antimicrobial activities, could be an interesting compound for septic shock treatment.

Biochem Pharmacol, 2003 Nov 1, 66(9), 1809 - 19
Chemically modified tetracyclines selectively inhibit IL-6 expression in osteoblasts by decreasing mRNA stability; Kirkwood K et al.; In bone biology, interleukin (IL)-6 is an autocrine/paracrine cytokine which can induce osteoclasts formation and activation to help mediate inflammatory bone destruction . Previous studies have shown that tetracycline and its derivatives have potentially beneficial therapeutic effects in the prevention and treatment of metabolic bone diseases by modulating osteoblast and osteoclast activities . Our previous studies indicated that non-antimicrobial chemically modified tetracyclines (CMTs) can dose-dependently inhibit IL-1 beta-induced IL-6 secretion in osteoblastic cells . In the present study, we explored the molecular mechanisms underlying the ability of doxycycline analogs CMT-8 and its non-chelating pyrazole derivative, CMT-5 to affect IL-6 gene expression in murine osteoblasts . Steady-state IL-6 mRNA was decreased with CMT-8 (ca . 50%) but not by CMT-5 when stimulated by IL-1 beta . CMT-8 regulation of IL-1 beta-induced IL-6 gene expression was further explored . CMT-8 did not affect IL-6 promoter activity in reporter gene assays . However, the IL-6 mRNA stability was decreased in the presence of CMT-8 . These effects require de novo protein synthesis as they were inhibited by cycloheximide . Western blot analysis indicated that CMT-8 did not affect p38 mitogen-activated protein kinase, c-jun NH(2)-terminal kinases, or extracellular signal-regulated kinases (1 and 2) phosphorylation in response to IL-1 beta . These data suggest that CMT-8 can modulate inhibit IL-1 beta-induced IL-6 expression in MC3T3-E1 cells at the post-transcriptional level affecting IL-6 mRNA stability . These observations may offer a novel molecular basis for this treatment of metabolic bone diseases that are mediated by IL-6.

Insect Biochem Mol Biol, 2003 Nov, 33(11), 1155 - 64
Proventriculus (cardia) plays a crucial role in immunity in tsetse fly (Diptera: Glossinidiae); Hao Z et al.; Fat body and hemocytes play a central role in cellular and humoral responses for systemic infections in invertebrates, similar to the mammalian liver and blood cells . Epithelial surfaces, in particular the midgut, participate in the initial local immune responses in order to aid in the generation of the terminal cytotoxic molecules that mediate non-self recognition . Here, we describe for the first time the immune responses of a cluster of cells at the foregut/midgut junction--known as proventriculus (cardia) in the medically and agriculturally important insect, tsetse fly (Diptera: Glossinidae) . We provide evidence for the transcriptional induction of the antimicrobial peptides attacin and defensin as well as for the reactive nitrogen intermediate (RNI) nitric oxide synthase (NOS) upon microbial challenge by either microinjection or feeding . Proventriculus from immune challenged flies also has higher NOS and nitric oxide (NO) activities as well as increased levels of the reactive oxygen intermediate (ROI), hydrogen peroxide (H2O2) . In several vector pathogen systems, including tsetse flies and African trypanosomes, stimulation of systemic responses prior to pathogen acquisition has been shown to reduce disease transmission . Furthermore, the induction of systemic immune responses has been documented while pathogens are still differentiating within the midgut environment . While evidence for a close molecular communication between the local and systemic responses is accumulating, the molecular signals that mediate these interactions are at present unknown . Reactive intermediates such as NO or H2O2 may function as immunological signals for mediating the molecular communication between the different insect compartments . We discuss the putative role of the proventriculus in invertebrate immunity and specifically speculate on its significance for trypanosome transmission in tsetse.

Int J Infect Dis, 2003 Sep, 7(3), 198 - 205
Infections with rapidly growing mycobacteria: report of 20 cases; Sungkanuparph S et al.; OBJECTIVES: A series of cases infected with rapidly growing mycobacteria was studied to determine the spectrum of disease, antimicrobial susceptibility, treatment, and outcome . METHODS: The cases identified as infections with rapidly growing mycobacteria in Ramathibodi Hospital from January 1993 to December 1999 were retrospectively studied . RESULTS: Most of the cases had no underlying disease . Only two cases were HIV-infected patients . The presenting clinical features were lymphadenitis (seven cases), skin and/or subcutaneous abscess (seven cases), localized eye infection (four cases), pulmonary infection (one case), and chronic otitis media (one case) . Four of seven cases with lymphadenitis had Sweet's syndrome, and one had psoriasis as an associated skin manifestation . Anemia was present in five cases, and improved with treatment of the primary disease . The organisms were Mycobacterium chelonae/abscessus group (17 cases) and Mycobacterium fortuitum group (three cases) . Susceptibility patterns of the organisms showed susceptibility to amikacin, netilmicin, and imipenem . M . fortuitum group was susceptible to more antibiotics than M . chelonae/abscessus group . The clinical responses corresponded to the antimicrobial susceptibility . Combinations of two or more drugs were used for the medical treatment . Surgical resection was performed where possible, to reduce the load of the organism, especially in cases with very resistant organisms . CONCLUSIONS: Infections with rapidly growing mycobacteria can occur in apparently normal hosts . The clinical syndrome is variable . The pathology is nonspecific . Clinical responses varied, but seemed to correlate with the in vitro susceptibility result . More studies are needed to enable us to deal with this infection effectively.

Avian Dis, 2003 Jul-Sep, 47(3), 588 - 93
In vitro antibiotic resistance profiles of Ornithobacterium rhinotracheale strains from Minnesota turkeys during 1996-2002; Malik YS et al.; Antimicrobial resistance in nearly all human and animal pathogens is on the increase . In poultry, Ornithobacterium rhinotracheale has been identified as a newly emerging respiratory bacterial pathogen that has caused significant economic losses to the poultry industry . In this study, we examined in vitro antibiotic resistance profiles of 125 isolates of O . rhinotracheale isolated from turkeys in Minnesota during 1996-2002 . A majority of isolates was sensitive to clindamycin, erythromycin, spectinomycin, and ampicillin . Resistance against sulfachloropyridiazine decreased from 1996 to 2002, but an increase in resistance was seen against gentamicin, ampicillin, trimethoprim sulfa, and tetracycline . The annual trend slopes for these antibiotics were 7.36%, 3.02%, 2.43%, and 1.95%, respectively . The resistance against penicillin remained constant from year to year with a trend slope of only 0.54% per year . These results emphasize the need for continued monitoring of O . rhinotracheale isolates for antibiotic resistance and establishment of baseline resistance pattern data for this organism . These data can then be used to design and evaluate local epidemiological interventions.

J Biochem (Tokyo), 2003 Sep, 134(3), 473 - 8
Cloning and analysis of the beta-lactamase gene from epsilon-poly-L-lysine-producing actinomycete Streptomyces albulus IFO14147; Hoshino Y et al.; Streptomyces albulus IFO14147 produces epsilon-poly-L-lysine, which exhibits antimicrobial activity . It is necessary for its molecular breeding to develop host-vector systems . We recently found a novel cryptic plasmid, pNO33, in this strain . As part of a search for a selectable marker gene for pNO33, we report here the isolation and analysis of the beta-lactamase gene of this strain, which can grow on ampicillin-containing plates . It was shown that the beta-lactamase production in S . albulus was induced by ampicillin . By introducing a genomic library of S . albulus into Escherichia coli, a 3.6-kbp fragment was identified as the region involved in ampicillin resistance . It contained three open reading frames, all of which are highly homologous to the beta-lactamase (the blaL product) and its regulatory proteins (the blaA and blaB products) of S . cacaoi . The growth phenotypes and enzyme assaying of E . coli and S . lividans showed that the blaL homologue (blaSa) encodes a beta-lactamase required for ampicillin resistance . The beta-lactamae gene can be utilized as a selectable marker in a cloning vector of S . albulus . However, the beta-lactamase activity was decreased in E . coli and repressed in S . lividans by the blaA and blaB homologues (blaASa and blaBSa) . It appears as if the blaASa product is a repressor of blaSa instead of an activator as in S . cacaoi.

Protein Pept Lett, 2003 Oct, 10(5), 497 - 502
Factors determining the efficacy of alpha-helical antimicrobial peptides; Dennison SR et al.; A database of alpha-helical antimicrobial peptides (AMP) was established and their minimum inhibitory concentrations (MIC) were compared with their physiochemical characteristics in an attempt to establish those features that determine efficacy . There is no significant difference in AMP sensitivity between Gram-positive and Gram-negative bacteria but fungi did require higher concentrations to achieve the same degree of growth inhibition . For antibacterial peptides there appears to be a positive correlation between MIC and hydrophobic arc size and a negative correlation between MIC and net charge.

Ann Chir, 2003 Sep, 128(7), 438 - 46
{Long-term usefulness of an information programme on practices in surgical antimicrobial prophylaxis}; Lallemand De Conto S et al.; INTRODUCTION: In France, numerous concordant studies show that there are some discrepancies between guidelines on surgical antibiotic prophylaxis and the current practice . In a previous study, conducted in April-June 2001, we found that the rate of appropriateness of surgical antimicrobial prophylaxis was approximately 40% . An information programme was implemented and the purpose of this paper was to present the short- and long-term usefulness of this campaign . METHODS: A total of 13 pairs of surgeons/anaesthetists participated in data collection during the three periods of the study . Prescriptions were observed in order to answer to five questions . Five variables describing practices concerning antibiotic prophylaxis in surgery were compared to national recommendations (updated in 1999): did the surgical procedure require antibiotic prophylaxis and was this carried out? Was the antibiotic used appropriate? Was the timing of the first injection optimal? Was the total duration of the treatment correct? Was the dose correct? RESULTS: The overall compliance with recommendations was significantly improved during the third period (P = 0.0002) . This improvement was particularly marked for antimicrobial prophylaxis duration . CONCLUSION: It seems that sequential surveillance of antimicrobial prophylaxis, including numerous surgical teams, could considerably improve the practices, if it was associated to informations that allowed physicians to appropriate the procedures.

Curr Opin Pharmacol, 2003 Oct, 3(5), 513 - 9
Targeting virulence for antimicrobial chemotherapy; Lee YM et al.; Untreatable bacterial infections constitute a dark but valid threat, with numbers of antibiotic resistant pathogens, as well as newly emerging ones, rising quickly . To combat this dangerous prospect, growing research into antimicrobials could be aimed at targeting the virulence of pathogens . Virulence refers to an organism's ability to establish an infection and cause disease . Many steps involved in the infection process can be targeted, including adherence, invasion and host defense evasion . Identification and characterization of virulence factors that aid in bacterial pathogenicity will lead to new drugs that can be applied to a variety of pathogens.

Curr Opin Pharmacol, 2003 Oct, 3(5), 508 - 12
Transcriptional profiling and drug discovery; Shaw KJ et al.; The availability of whole-genome nucleotide sequence data from an ever-growing list of microbial genomes, including complete genomes for multiple strains within a species, presents an opportunity to overcome the challenges presented by antimicrobial drug resistance . The development of DNA microarrays provides a unique tool to understand pathogenic microbial genomes from a global perspective . Genome-wide expression profiles can facilitate the characterization both of the mechanisms of action and of the mechanisms of resistance to antimicrobial agents . Expression data have also been used to initiate the characterization of genes of unknown function, potentially leading to the identification of novel drug targets . Initial studies using DNA microarrays to analyse the host and pathogen responses to infection have also been performed, impacting our understanding of pathogenesis and the strategies taken to combat infectious diseases.

Curr Opin Pharmacol, 2003 Oct, 3(5), 470 - 3
The abandonment of antibacterials: why and wherefore?
Shlaes DM.
The pharmaceutical industry is currently abandoning its antibacterial discovery research efforts . This seems to be part of a cyclical pattern in this therapeutic area . The reasons behind these ongoing cycles of feast and famine are multiple, but most revolve around the perception of market opportunities from the continuing emergence of resistance, balanced against the difficulties in the discovery of novel antibacterial compounds, the costs of development and the general regulatory and financial environment in which companies find themselves . Relief for the industry will require both regulatory and legislative action at a time when this will be politically difficult to achieve . In the meantime, the problems of antimicrobial resistance are not going away.

Curr Opin Pharmacol, 2003 Oct, 3(5), 459 - 63
Do we really need new anti-infective drugs?
Rice LB.
Nosocomial infections continue to be important causes of morbidity and mortality in modern hospitals . The large volume and often excessive use of antimicrobial agents, particularly in critical care settings, has led to the emergence of pathogens that are resistant to virtually all available antimicrobial agents . Moreover, patients infected with organisms that are susceptible to one or more agents might not tolerate those agents, leaving physicians with no therapeutic options . Finally, the need to move patients out of the hospital, and thereby avoid nosocomial infections, has created an ever-growing need for antimicrobial agents whose pharmacokinetic profiles favour a single daily dosing that can be administered at home.

J Pak Med Assoc, 2003 Aug, 53(8), 328 - 32
Frequency and antibiotic susceptibility pattern of mycobacterial isolates from extra-pulmonary tuberculosis cases; Butt T et al.; OBJECTIVE: To determine the frequency and antimicrobial susceptibility pattern of extra-pulmonary tuberculosis in Rawalpindi . SETTING: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi . METHODS: Between September 2000 and August 2002, 899 pulmonary and 460 extra-pulmonary specimens from suspected cases of tuberculosis were examined . The radiometric BACTEC 460 TB system was used for culture and antimicrobial susceptibility testing . RESULTS: Mycobacteria were isolated from 291 pulmonary specimens and 98 extra-pulmonary specimens . The frequency of extra-pulmonary tuberculosis was 25.2% . The commonest source of isolation was pus (44.9%, frequency 11.3%), followed by lymph nodes (13.3%, frequency 3.3%) and pleural fluid (13.3%, frequency 3.3%) . We tested the anti-microbial susceptibility of the isolates to the four first line anti-tuberculous drugs, rifampicin, isoniazid, streptomycin and ethambutol . Of the extra-pulmonary isolates 13.3% were resistant to a single drug, 21.4% were multi-drug resistant and 9.2% were resistant to all the four drugs . CONCLUSION: Increased awareness of the magnitude of the problem posed by extra-pulmonary tuberculosis is required so that appropriate control measures can be adopted.

Rev Med Chil, 2003 Aug, 131(8), 847 - 56
{Medical outcomes and antimicrobial compliance according to the Chilean Society of Respiratory Diseases guidelines for hospitalized patients with community acquired pneumonia}; Diaz A et al.; BACKGROUND: The Chilean Society of Respiratory Diseases (SER) developed guidelines for the empirical treatment of community acquired pneumonia (CAP) . AIM: To evaluate the degree of adherence to antibiotic treatment recommended by SER guidelines and its influence on medical outcomes . PATIENTS AND METHODS: We prospectively evaluated 453 consecutive immunocompetent adults (mean age +/- SD: 69 +/- 19 years) hospitalized for CAP . Patients were stratified according to the Pneumonia Severity Index (PSI), and initial antibiotic regimen was classified as being consistent or inconsistent with the SER guidelines . Rate of medical complications, switch therapy rate, length of stay (LOS), and 30 days mortality were compared between those treated consistently or inconsistently with the SER guidelines . RESULTS: Adherence to SER guidelines was 46% . Patients treated consistently with the SER guidelines were older (mean age +/- SD: 72 +/- 16 v/s 65 +/- 20 years), had more comorbidities (84 v/s 69%) and a higher proportion belonged to the high-risk PSI categories (69 v/s 49%) . There were no significant differences in medical complication rate, switch therapy rate or LOS between both groups . Adherence to SER guidelines did not affect mortality after adjusting for PSI and for prognostic factors associated with 30 days mortality by multivariate analysis . CONCLUSIONS: The degree of adherence to antibiotic treatment recommended by SER guidelines was moderate and they were applied mainly in patients with high risk CAP . This fact can explain the lack of evidence of improved medical outcome in patients treated according to SER guidelines.

Clin Infect Dis, 2003 Nov 1, 37(9), 1201 - 9 Epub 2003 Sep 30.
Hypermutation as a factor contributing to the acquisition of antimicrobial resistance; Blazquez J; Contrary to what was thought previously, bacteria seem to be, not merely spectators to their own evolution, but, through a variety of mechanisms, able to increase the rate at which mutations occur and, consequently, to increase their chances of becoming resistant to antibiotics . Laboratory studies and mathematical models suggest that, under stressful conditions, such as antibiotic challenge, selective pressure favors mutator strains of bacteria over nonmutator strains . These hypermutable strains have been found in natural bacterial populations at higher frequencies than expected . The presence of mutator strains in the clinical setting may indicate an enhanced risk of acquiring antibiotic resistance through mutational and recombinational events . In addition, some antibiotics are inducers of mechanisms that transiently increase the mutation rate, and thus probably act, not only as mere selectors of antibiotic resistant clones, but also as resistance-promoters.

Parasitol Res, 2003 Dec, 91(6), 476 - 81 Epub 2003 Oct 14.
Up-regulated humoral immune response in the soft tick, Ornithodoros moubata (Acari: Argasidae); Nakajima Y et al.; Ticks have an efficient defense system for preventing microbial infection . The antimicrobial peptide defensin is one effective molecule in this system . Here we investigated immune competence and the involvement of defensin in the humoral defense of the soft tick, Ornithodoros moubata . Semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that gene expression of all four defensin isoforms was up-regulated by bacteria or bacterial components . Defensin gene up-regulation by hemocoelic inoculation of bacteria involves the midgut and granulocytes . In immunodetection analysis, immunization by bacterial injection increases the relative concentration of defensin-like material in the hemolymph plasma . Furthermore, elevated antibacterial activity against Gram-positive bacteria but not against Gram-negative bacteria was observed after immunization by a liquid growth inhibition assay . Therefore, enhanced anti-Gram-positive bacterial activity appears to be partially dependent on the release of defensin into the hemolymph . These findings demonstrate that defensin plays an important role in the up-regulated humoral response of O . moubata.

Clin Microbiol Rev, 2003 Oct, 16(4), 597 - 621
Infections in patients with inherited defects in phagocytic function; Andrews T et al.; Patients with defects in phagocytic function are predisposed to intracellular microorganisms and typically have early dissemination of the infection . Recognition of the underlying disorder and aggressive antimicrobial therapy has been beneficial for the patients . Improved understanding of the pathophysiology has also affected patient management by allowing specific, targeted immunomodulatory intervention . The disorders described in this review are not common but have had a significant impact on our understanding of the role of phagocytic cells in host defense . Conversely, understanding the role of the neutrophil and macrophage in infection has benefited not just the patients described in this review but also other patients with similar disease processes.

Burns, 2003 Nov, 29(7), 702 - 10
A randomised clinical trial comparing a hydrocolloid-derived dressing and glycerol preserved allograft skin in the management of partial thickness burns; Vloemans AF et al.; Membranous dressings for the treatment of partial and mixed thickness burns are among the most innovative and promising new developments of the last years . In this study, we present data of a randomised prospective comparative study on a carboxymethylcellulose based dressing, Hydrofibre((R)) and glycerolized human allograft skin.In a 2 year period, 80 patients (40 for each material) were enrolled in the trial . Study wounds (<10% TBSA) that had not re-epithelialised after 14+/-3 days were debrided and grafted or, if small enough, managed with a topical antimicrobial agent . Mean total TBSA was 8.3+/-5.2%, study burn 3.7+/-2.0% for the Hydrofibre((R)) group and 7.3+/-4.3% total, 3.4+/-2.1% study burn for the allograft skin group (n.s . Wilcoxon rank sum test) . No significant differences between groups were established in number of patients with superficial/deep burns.In both groups about 2/3 of the patients healed completely with the dressings applied (24/40 versus 27/40 for Hydrofibre((R)) versus allograft skin, respectively) . However, a higher incidence of post-study excision and grafting was found in the Hydrofibre((R)) group (45% versus 15% in the allograft skin group, P=0.004, Mann-Whitney) . At 10 weeks follow-up no significant differences were seen in scar colour, pigmentation, pliability, height or itching (Vancouver Scar Scale) . Skin elasticity, measured by the Cutometer((R)), was significantly better for the allograft group (P=0.010, Wilcoxon) . These differences were no longer found at 6 months and 1 year follow-up . Incidence of hypertrophy after 6 months was higher, but not significantly, in the Hydrofibre((R)) compared to the allograft skin group (52.5% versus 30%, P=0.09, chi-square).In view of the results from our comparative study on Hydrofibre((R)) versus allograft skin, we prefer the use of allograft skin for the category of larger burns of mixed depth, usually presented to burn centres . However, for partial thickness and small burns Hydrofibre((R)) can be the first choice in treatment.

Eye Contact Lens, 2003 Oct, 29(4), 258 - 61
Doxycycline in the management of pseudomonas corneal melting: two case reports and a review of the literature; McElvanney AM; BACKGROUND: Pseudomonas keratitis can result in the breakdown of collagen with subsequent corneal melting and perforation . The use of antimetalloproteinases may help to stabilize melting and prevent imminent perforation of the cornea . The use of topical protease inhibitors and neutrophil inhibitors is of limited value . Tetracyclines, however, have been shown to have anticollagenolytic activity and inhibit metalloproteinases, and they may suppress connective tissue breakdown . PURPOSE: To establish the stabilization of corneal melting in cases of Pseudomonas keratitis . METHODS: Two young patients with severe contact lens-associated Pseudomonas keratitis and corneal melting were treated with oral doxycycline and standard topical treatment . RESULTS: Corneal melting was stabilized in each patient, and perforation was avoided . CONCLUSIONS: Tetracyclines have an anticollagenolytic action in addition to their antimicrobial activity . The use of doxycycline as an adjunctive therapy in the management of Pseudomonas corneal melting may help to stabilize corneal breakdown and prevent subsequent perforation.

Eye Contact Lens, 2003 Oct, 29(4), 245 - 9
Comparative antimicrobial activity of no-rub multipurpose lens care solutions in the presence of organic soil; McGrath D et al.; PURPOSE: This study reports the effect of organic soil on the antimicrobial activity of four commercially available multipurpose contact lens care solutions used in no-rub regimens as determined by a modified International Organization for Standardization 14729 (ophthalmic optics-contact lens care products-microbiological requirements for products and regimens for hygienic management of contact lenses) Stand Alone Test procedure . METHODS: Testing was performed with organic soil consisting of a mixture of heat-killed yeast cells and heat-inactivated bovine serum or these components added separately . The organic soil was mixed with the challenge microorganisms before addition to the solution, added to the solution after addition of the challenge microorganisms, or added to the solution before addition of the challenge microorganisms . A final concentration of 0.4% v/v organic soil and 1 x 105 to 1 x 106 cfu/mL challenge microorganisms was realized in all test cases . RESULTS: The antimicrobial activity of no-rub multipurpose lens care solutions was reduced in the presence of added organic soil . The extent of reduction varied with the type of organic soil, the method of addition of the organic soil to the solution, the challenge organism, and the solution tested . CONCLUSION: The type of organic soil used and the method of addition of organic soil to the lens care solution may affect antimicrobial activity as determined by the primary criteria of the International Organization for Standardization Stand Alone Test procedure . The overall reduction in antimicrobial activity depends on the solution and organism.

Bioorg Med Chem Lett, 2003 Nov 3, 13(21), 3771 - 3
New benzylidenethiazolidinediones as antibacterial agents; Heerding DA et al.; A novel benzylidenethiazolidinedione has been discovered with antimicrobial activity . Here, we present the results of a structure-activity study on this compound with respect to its antimicrobial activity.

Braz J Infect Dis, 2003 Oct, 7(5), 290 - 6
Effectiveness of the actions of antimicrobial control in the intensive care unit; Dos Santos EF et al.; There are various strategies to improve the effectiveness of antibiotics in hospitals . In general, for the implementation of guidelines for appropriate antibiotic therapy, the participation of infectious disease (ID) physicians deserves considerable attention . This study was a prospective ecological time-series study that evaluates the effectiveness of the ID physician's opinion to rationalize and control the use of antibiotics in medical-surgical intensive care units (ICU), and the impact of their intervention on treatment expenditures . There was significant change in the pattern of use of antimicrobials, this pattern approximating that of a medical-surgical ICU that participates in the ICARE (Intensive Care Antimicrobial Resistance Epidemiology) Project . For example, there was a significant increase in the consumption of antimicrobials of the ampicillin group (Relative Risk {RR}=3.39; 95% CI: 2.34-4.91) and antipseudomonal penicillins (RR=2.89; 95% CI: 1.70-4.92) . On the other hand, there was a significant reduction in the consumption of 3(rd)/4(th )generation cephalosporins (RR=0.66; 95% CI: 0.57-0.77) and carbapenems (RR=0.43; 95% CI: 0.33-0.56) . On average, for every patient-day antibiotic expense was reduced 37.1% during calendar year 2001, when compared with 2000 . The ID specialists' opinion and the adoption of guidelines for empirical antibiotic therapy of hospital-acquired pneumonia contributed to a reduction in the use of antimicrobials in medical-surgical ICU . However, further studies that have more control over confounding variables are needed to help determine the relevance of these discoveries.

Blood, 2004 Feb 15, 103(4), 1534 - 41 Epub 2003 Oct 09.
Transfer of allogeneic CD62L- memory T cells without graft-versus-host disease; Chen BJ et al.; The major challenge in allogeneic hematopoietic cell transplantation is how to transfer allogeneic T-cell immunity without causing graft-versus-host disease (GVHD) . Here we report a novel strategy to selectively prevent GVHD by depleting CD62L(+) T cells (naive and a subset of memory T cells) . In unprimed mice, CD62L(-) T cells (a subset of memory T cells) failed to proliferate in response to alloantigens (which the mice have never previously encountered) and were unable to induce GVHD in allogeneic hosts . CD62L(-) T cells contributed to T-cell reconstitution by peripheral expansion as well as by promoting T-cell regeneration from bone marrow stem/progenitor cells . CD62L(-) T cells from the animals previously primed with a tumor cell line (BCL1) were able to inhibit the tumor growth in vivo but were unable to induce GVHD in the third-party recipients . This novel technology may allow transfer of allogeneic recall antitumor and antimicrobial immunity without causing GVHD.

Biotechnol Adv, 2001 Jul, 19(4), 299 - 316
The prospects of modifying the antimicrobial properties of milk; Kolb AF; Milk contains a variety of substances, which inhibit the infection of pathogens . This is of benefit to the mother, safeguarding the integrity of the lactating mammary gland, but also of huge importance for protection of the suckling offspring . The antimicrobial substances in milk can be classified into two categories . First, nonspecific, broad-spectrum defense substances, which have evolved over long periods of time, and secondly, substances like antibodies, which are specifically directed against particular pathogens and have developed during the mother's lifetime . Substances in both categories may be targets for biological intervention and manipulation with the goal of improving the antimicrobial properties of milk . These alterations of milk composition have applications in human as well as in animal health.

Helicobacter, 2003, 8(5), 542 - 53
Inhibition of Helicobacter pylori-induced nuclear factor-kappa B activation and interleukin-8 gene expression by ecabet sodium in gastric epithelial cells; Kim JM et al.; BACKGROUND: Helicobacter pylori stimulates nuclear factor-kappa B (NF-kappa B) activation and chemokine interleukin-8 (IL-8) expression in gastric epithelial cells . Ecabet sodium (ecabet), a locally acting antiulcer drug, is known to have anti-H . pylori activity . However, there is little understanding of how ecabet induces anti-inflammatory activity in gastric epithelial cells infected with H . pylori . The aim of this study was to investigate the effects of ecabet on IL-8 gene expression and NF-kappa B activation in human gastric epithelial cells infected with H . pylori . MATERIALS AND METHODS: After Hs746T, MKN-45, or SNU-5 gastric epithelial cell lines had been infected with cagA+cytotoxin+H . pylori in the presence of ecabet, IL-8 mRNA expression was assessed by quantitative reverse transcription-polymerase chain reaction, and IL-8 secretion was measured by enzyme-linked immunosorbent assay . NF-kappa B and inhibitory kappa B-alpha (I kappa B alpha) signals were assayed by electrophoretic mobility shift assay and Western blot, respectively . The activation of NF-kappa B and IL-8 reporter genes was determined by luciferase assay . RESULTS: Ecabet showed no antimicrobial activiy against Gram-positive or -negative bacteria . However, ecabet inhibited transcription of the IL-8 gene and secretion of IL-8 by gastric epithelial cells infected with H . pylori at a concentration of 5 micro g/ml . Moreover, ecabet inhibited the activation of NF- kappa B and the degradation of I kappa B alpha in gastric epithelial cells in response to H . pylori infection . In addition, the NF-kappa B signal inhibited by ecabet was comprised predominantly of heterodimers of p65/p50 . CONCLUSIONS: Ecabet inhibited H . pylori-induced IL-8 gene transcription and secretion by suppressing the NF-kappa B signal . This inhibition might be one pathway by which ecabet exerts its anti-inflammatory effect on H . pylori-induced gastric inflammation.

Aliment Pharmacol Ther, 2003 Oct 15, 18(8), 821 - 7
Role of antibiotic sensitivity testing before first-line Helicobacter pylori eradication treatments; Neri M et al.; BACKGROUND: The resistance of Helicobacter pylori to antibiotics has been advocated as a major cause of treatment failure, and antimicrobial sensitivity testing has been proposed to improve efficacy; however, its role before first-line therapy has not been investigated in detail . AIM: To assess whether antimicrobial sensitivity testing improves the eradication rate of first-line anti-Helicobacter treatments and to compare the effectiveness of ranitidine bismuth citrate and omeprazole in the presence of H . pylori resistance to antibiotics . METHODS: Two hundred and forty-two patients were assigned to either empirical or antimicrobial sensitivity testing-based treatment; within each group, subjects were further randomized to receive ranitidine bismuth citrate, 400 mg b.d., tinidazole, 500 mg b.d., and clarithromycin, 500 mg b.d., or omeprazole, 20 mg b.d., clarithromycin, 500 mg b.d., and amoxicillin, 1 g b.d., for 1 week, with substitution of the resistant antibiotic in the antimicrobial sensitivity testing-based treatment group . RESULTS: Eradication rates were 67% {confidence interval (CI), 55-79%} in the empirical treatment group and 76% (CI, 65-87%) in the antimicrobial sensitivity testing-based group (P=N.S.) . The overall success rate was 60% (CI, 51-69%) with omeprazole and 82% (CI, 73-91%) with ranitidine bismuth citrate (P<0.03); the latter overcame antibiotic resistance in 12 of 15 strains vs . zero of eight strains by omeprazole . CONCLUSIONS: Antimicrobial sensitivity testing before first-line treatment does not improve the eradication rate, which is greater when ranitidine bismuth citrate is included in the treatment.

Aliment Pharmacol Ther, 2003 Oct 15, 18(8), 799 - 804
Esomeprazole-based one-week triple therapy with clarithromycin and metronidazole is effective in eradicating Helicobacter pylori in the absence of antimicrobial resistance; Miehlke S et al.; AIM: This study aimed to investigate the effectiveness of a one-week triple therapy with esomeprazole, clarithromycin and metronidazole for eradication of Helicobacter pylori infection in the absence of antimicrobial resistance . METHODS: Patients testing positive for H . pylori susceptible to metronidazole and clarithromycin (E-test) were randomized to receive a one-week regimen with either esomeprazole 2 x 20 mg or omeprazole 2 x 20 mg in combination with clarithromycin 2 x 250 mg and metronidazole 2 x 400 mg . Follow-up endoscopy with histology and culture and/or rapid urease test was performed 4-8 weeks after the end of treatment . RESULTS: Eighty patients were randomized . Helicobacter pylori infection was cured in 38/39 patients of the esomeprazole group and 31/33 patients of the omeprazole group (per protocol 97.4% (95% confidence interval {CI}, 86.2-99.9), 93.7% (95% CI, 79.2-99.2), P=0.59); intention-to-treat 90.4% (95% CI: 77.4-97.3), 81.6% (95% CI: 65.7-92.3), respectively . No major side effects occurred . Minor side effects occurred in eight (20%) and six (23%) patients during esomeprazole and omeprazole therapy, respectively . Post-treatment susceptibility testing revealed resistance to both metronidazole and clarithromycin in two of the three patients who failed . CONCLUSION: We conclude that esomeprazole, clarithromycin and metronidazole as one-week triple therapy is effective for eradication of H . pylori in the absence of antimicrobial resistance.

Andrologia, 2003 Oct, 35(5), 252 - 7
Epidemiology and demographics of prostatitis; Schaeffer AJ; Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a multifactorial problem affecting men of all ages and demographics . Currently, there is a relative dearth of epidemiological information on CPPS . It is clear that patients with CPPS have a dismal quality of life and many have benefited only minimally from empiric, goal-directed therapy . Long-term follow-up of the CPPS cohort will answer important questions about the natural and treated history of this syndrome . Similarly, ongoing and future studies will provide community-based and prevalence estimates for CPPS, morbidity rates for men with CPPS, and the rates of symptom improvement and symptom deterioration for these men, as well as the probability of benefits and harm from different treatments . Although men with CP routinely receive anti-inflammatory and antimicrobial therapy, recent studies suggest that leucocyte and bacterial counts do not correlate with severity of symptoms . These findings suggest that factors other than leucocytes and bacteria contribute to the symptoms associated with CPPS . The probability of benefits and harm from different treatments for CPPS, and reliable and valid measures to define these outcomes are eagerly awaited.

J Org Chem, 2003 Oct 17, 68(21), 8185 - 92
Synthesis of orthogonally protected lanthionines; Mohd Mustapa MF et al.; Synthetic approaches to the lantibiotics, a family of thioether-bridged antimicrobial peptides, require flexible synthetic routes to a variety of orthogonally protected derivatives of lanthionine 1 . The most direct approaches to lanthionine involve the reaction of cysteine with an alanyl beta-cation equivalent . Several possibilities exist for the alanyl beta-cation equivalent, including direct activation of serine under Mitsunobu conditions: however, the low reactivity of sulfur nucleophiles in the Mitsunobu reaction has previously precluded its use in the synthesis of the lantibiotics . We report here a new approach to the synthesis of protected lanthionine, using a novel variant of the Mitsunobu reaction in which catalytic zinc tartrate is used to enhance the nucleophilicity of the thiol . In the course of these studies, we have also demonstrated that the synthesis of lanthionine from trityl-protected beta-iodoalanines is prone to rearrangement, via an aziridine, to give predominantly trityl-protected alpha-iodo-beta-alanines, and hence norlanthionines, as the major products.

Arq Gastroenterol, 2003 Jan-Mar, 40(1), 55 - 60 Epub 2003 Oct 06.
The impact of Helicobacter pylori resistance on the efficacy of a short course pantoprazole based triple therapy; Eisig JN et al.; BACKGROUND: Many of the currently used Helicobacter pylori eradication regimens fail to cure the infection due to either antimicrobial resistance or poor patient compliance . Those patients will remain at risk of developing potentially severe complications of peptic ulcer disease . AIM: We studied the impact of the antimicrobial resistance on the efficacy of a short course pantoprazole based triple therapy in a single-center pilot study . METHODS: Forty previously untreated adult patients (age range 20 to 75 years, 14 males) infected with Helicobacter pylori and with inactive or healing duodenal ulcer disease were assigned in this open cohort study to 1 week twice daily treatment with pantoprazole 40 mg, plus clarithromycin 250 mg and metronidazole 400 mg . Helicobacter pylori was assessed at entry and 50 3 days after the end of treatment by rapid urease test, culture and histology of gastric biopsies . The criteria for eradication was a negative result in the tests . Susceptibility of Helicobacter pylori to clarithromycin and metronidazole was determined before treatment with the disk diffusion test . RESULTS: One week treatment and follow up were complete in all patients . Eradication of Helicobacter pylori was achieved in 35/40 patients (87.5%) and was higher in patients with nitroimidazole-susceptible strains {susceptible: 20/20 (100%), resistant: 10/15 (67%)} . There were six (15%) mild adverse events reports . CONCLUSIONS: A short course of pantoprazole-based triple therapy is well tolerated and effective in eradicating Helicobacter pylori . The baseline metronidazole resistance may be a significant limiting factor in treatment success.

Folia Microbiol (Praha), 2003, 48(4), 543 - 7
Activation of human leukocytes by lipid A from E . coli strains adapted to quaternary ammonium salt and amine oxide; Dubnickova M et al.; The immunomodulatory activities of monophosphoryl lipid A (MLA) and diphosphoryl lipid A analogues obtained from the sensitive strain of E . coli and from the resistant strains adapted to a quaternary ammonium salt and an amine oxide were compared . All analogues considerably stimulated the activity of human leukocytes although the analogue from the sensitive strain at a higher concentration significantly suppressed phagocytosis . The MLA analogue exhibited a suppressive effect on the microbicidal activity of human leukocytes against E . coli and the peroxidase activity . Adaptation of bacteria to amphiphilic antimicrobial compounds, which is accompanied by chemical changes in their lipid A, only slightly reduced their immunomodulatory activity when compared with the analogue from the sensitive strain . On the other hand, the diphosphoryl analogues were less active than MLA.

Aust Crit Care, 2003 Aug, 16(3), 101 - 10
Oral care of the critically ill: a review of the literature and guidelines for practice; O'Reilly M; Maintaining oral health in the critically ill patient is imperative in reducing the risk of nosocomial infections and improving patient comfort and discharge outcomes . Critically ill patients are at great risk for poor oral health as many are elderly, undernourished, dehydrated, immunosuppressed, have a smoking or alcohol history, are intubated or on high-flow oxygen, and are unable to mechanically remove dental plaque . Many modalities for delivering oral care have been reported in the literature . The use of the toothbrush in the mechanical removal of plaque, even in the intubated patient, has been proven to be superior to the swab . Brushing of the gums in edentulous patients is of benefit . Although electric toothbrushes are preferable, their cost, size and the potential for cross-infection limits their use . Chlorhexidine has long been the gold standard for mouthwashes and provides up to 24 hours of antimicrobial activity; therefore infrequent applications are adequate . Sodium bicarbonate and hydrogen peroxide are of limited use due to lack of convincing evidence regarding their safety and antimicrobial effects in the critically ill population . Saliva stimulants or substitutes including lemon and glycerine are also inappropriate for moistening the oral cavity in the critically ill patient . Regular oral assessment and individualized oral care, along with the use of a standardised protocol for oral care (incorporating proven modalities) is vital for optimal oral care in the critically ill patient.

J Clin Microbiol, 2003 Oct, 41(10), 4751 - 4
Direct susceptibility testing of positive blood cultures by using Sensititre broth microdilution plates; Chapin KC et al.; Traditional susceptibility testing of blood cultures requires overnight incubation in order to obtain isolated colonies . Susceptibility results can be reported up to 24 h sooner by using a bacterial pellet from the blood culture broth . This study evaluated the accuracy of direct susceptibility testing from positive ESP blood culture broths by using Sensititre broth microdilution plates compared to testing with isolated colonies . Practical inclusion criteria were applied to gram-positive organisms to avoid reporting susceptibilities for probable contaminants . All gram-negative organisms were tested directly . An aliquot of the blood culture was centrifuged, and the resulting pellet was used to make a 0.5 McFarland suspension . Microdilution plates were inoculated and interpreted according to the manufacturer's instructions . Colony counts were performed to ensure proper colony density was achieved . A total of 199 patient and seeded blood cultures were evaluated for both essential (within +/-1 twofold dilution) and categorical (sensitive, intermediate, or resistant) agreement . Testing of 93 gram-positive isolates (1,214 antimicrobial agent-organism combinations) yielded 98% essential agreement and categorical error rates of 0.3% minor, no major (false resistance), and 1.7% very major (false susceptibility) errors . For 106 gram-negative isolates (1,828 antimicrobial agent-organism combinations), the essential agreement was 99% . Categorical error rates were 0.5, 0, and 2.0% for minor, major, and very major errors, respectively . Performance was comparable for both gram-positive and gram-negative isolates, as well as for both aerobic and anaerobic media . Using this direct testing methodology, reliable susceptibility results can be reported to physicians 24 h sooner, allowing earlier appropriate modification of antimicrobial therapy.

Ann Rheum Dis, 2003 Nov, 62 Suppl 2, ii17 - 21
Roles of antimicrobial peptides such as defensins in innate and adaptive immunity; Oppenheim JJ et al.; A number of antimicrobial peptides such as defensins have multiple functions in host defence . Defensins are produced not only by phagocytic cells and lymphocytes, but also by the epithelial cell lining of the gastrointestinal and genitourinary tracts, the tracheobronchial tree, and keratinocytes . Some are produced constitutively, whereas others are induced by proinflammatory cytokines and exogenous microbial products . Defensins produced by cells in the course of innate host defence serve as signals which initiate, mobilise, and amplify adaptive immune host defences . Administration of defensins with antigens to mice enhances both cellular (Th1-dependent) and humoral (Th2-dependent) cytokine production and immune responses . Linkage of defensins to weak tumour antigens potentiates their immunoadjuvant effects . Defensins use multiple cellular receptors, which endows them with the capacity to marshall adaptive host defences against microbial invaders.

J Travel Med, 2003 Sep-Oct, 10(5), 290 - 2
Irrational prescribing in South Asia: a case of fluoroquinolone-associated phototoxicity; Cave W et al.; Prescribing habits in South Asian countries have been subjected to some scrutiny.1-6 Most studies conclude that the quality of prescribing is poor, with overuse of antimicrobials and irrational use of fixed-dose combination therapy, particularly in the private sector.1 Prescriptions for multiple drugs are the rule rather than the exception, with up to seven items being prescribed for a single disease entity . Analgesics, anti-inflammatories and drugs of uncertain pharmacologic efficacy, such as vitamins, minerals and glucose water, are also frequently prescribed.

J Am Chem Soc, 2003 Oct 15, 125(41), 12464 - 74
Microcin J25 has a threaded sidechain-to-backbone ring structure and not a head-to-tail cyclized backbone; Rosengren KJ et al.; Microcin J25 is a 21 amino acid bacterial peptide that has potent antibacterial activity against Gram-negative bacteria, resulting from its interaction with RNA polymerase . The peptide was previously proposed to have a head-to-tail cyclized peptide backbone and a tight globular structure (Blond, A., Peduzzi, J., Goulard, C., Chiuchiolo, M . J., Barthelemy, M., Prigent, Y., Salomon, R . A., Farias, R . N., Moreno, F . & Rebuffat, S . Eur . J . Biochem . 1999, 259, 747-755) . It exhibits remarkable thermal stability for a peptide of its size lacking disulfide bonds and in part this was previously proposed to derive from its macrocyclic structure . We show here that in fact the peptide does not have a head-to-tail cyclic structure but rather a side chain to backbone cyclization between Glu8 and the N-terminus . This creates an embedded ring that is threaded by the C-terminal tail of the molecule, forming a noose-like feature . The three-dimensional structure deduced from NMR data suggests that slippage of the noose is prevented by two aromatic residues flanking the embedded ring . Unthreading does not occur even when the molecule is enzymatically digested with thermolysin . The new structural interpretation fully accounts for previously reported NMR and biophysical data and is consistent with the remarkable stability of this potent antimicrobial peptide.

Pflugers Arch, 2004 Feb, 447(5), 571 - 9 Epub 2003 Oct 07.
SLC11 family of H+-coupled metal-ion transporters NRAMP1 and DMT1; Mackenzie B et al.; NRAMP1 (natural resistance-associated macrophage protein-1) and DMT1 (divalent metal-ion transporter-1) make up the SLC11 gene family of metal-ion transporters that are energized by the H(+) electrochemical gradient . Long known to confer resistance to bacterial infection, NRAMP1 functions at the phagolysosomal membrane of macrophages and neutrophils . NRAMP1 most likely contributes to macrophage antimicrobial function by extruding essential metal ions (including Mn(2+)) from the phagolysosome via H(+)/metal-ion cotransport . An alternative hypothesis in the literature proposes that NRAMP1 concentrate Fe(2+) within the phagolysosome by means of H(+)/Fe(2+) antiport, resulting in the generation of toxic free radicals . DMT1 is expressed widely and accepts as substrates a broad range of transition metal ions, among which Fe(2+) is transported with high affinity ( K(0.5) approximately 2 microM) . DMT1 accounts both for the intestinal absorption of free Fe(2+) and for transferrin-associated endosomal Fe(2+) transport in erythroid precursors and many other cell types . DMT1 is up-regulated dramatically in the intestine by dietary iron restriction and, despite high serum iron levels, is not appropriately down-regulated in hereditary hemochromatosis.

Curr Med Chem, 2003 Dec, 10(24), 2643 - 9
Mechanisms of action of DNA intercalating acridine-based drugs: how important are contributions from electron transfer and oxidative stress?
Baguley BC, Wakelin LP, Jacintho JD, Kovacic P.
Reactive oxygen species (ROS) are produced continuously in living cells as a by-product of respiration and other metabolic activity . Some ROS may react with DNA, and in some cases may abstract an electron from the double helix, leading to long range electron transfer (ET) reactions . Thus, the DNA of living cells may be in a continuous state of ET . We consider here whether acridine-based anticancer or antimicrobial drugs, which bind to DNA by intercalation, might either donate electrons to, or accept electrons from, the double helix, thus actively participating in ET reactions . We focus in particular on two acridine-based drugs that have been tested against human cancer in the clinic . Amsacrine is a 9-anilinoacridine derivative that appears to act as an electron donor in ET reactions on DNA, while N-{2-(dimethylamino)ethyl}acridine-4-carboxamide (DACA) may act as an electron acceptor . Such reactions may make important contributions to the antitumor activity of these drugs.

Nat Prod Res, 2003 Oct, 17(5), 375 - 80
Evaluation of the antiviral and antimicrobial activities of triterpenes isolated from Euphorbia segetalis; Madureira AM et al.; A phytochemical reinvestigation of the whole plant of Euphorbia segetalis yielded five tetracyclic triterpenes: 3beta-hydroxy-cycloart-25-en-24-one (1), cycloart-25-ene-3beta,24-diol (2), cycloart-23-ene-3beta,25-diol (3), lanosta-7,9(11),24-trien-3beta-ol (4) and lanosta-7,9(11),24(31)-trien-3beta-ol (5) . beta-acetoxy-cycloart-25-en-24-one (1a) and glutinol (6), lupenone (7), dammaranodienol (9), cycloartenol acetate (10), 24-methylenecycloartanol acetate (11) and beta-sitosterol (12), isolated previously, were evaluated for their antiviral activities against Herpes simplex virus (HSV) and African swine fever virus (ASFV) . Lupenone exhibited strong viral plaque inhibitory effect against HSV-1 and HSV-2 . The in vitro antifungal and antibacterial activities of la, cycloart-23-ene-3beta,25-diol, 3-acetate (3a) and 6-12 were also investigated.

Boll Chim Farm, 2003 Jun, 142(5), 232 - 8
Synthesis and biological evaluation of some new substituted naphthoquinones; Habib NS et al.; Two novel series of 1,4-naphthoquinone derivatives have been synthesized namely; N-ethoxycarbonyl-2-ethoxycarbonyloxy-3- alkyl-1,4-naphthoquinon-1-substituted phenylhydrazones 3a-f and 2-chlorocetyloxy-3-alkyl-1,4-naphthoquinone-1-substituted phenylhydrazones 4a-d . The antimicrobial activity as well as anticancer activity of these compounds have been evaluated . The acute toxicity of the active compounds was determined.

Berl Munch Tierarztl Wochenschr, 2003 Sep-Oct, 116(9-10), 353 - 61
{Antimicrobial susceptibility testing of bacteria isolated from animals: methods for in-vitro susceptibility testing and their suitability with regard to the generation of the most useful data for therapeutic applications}; Schwarz S et al.; In-vitro susceptibility testing provides valuable informations for choosing the most suitable antimicrobial agent for the control of bacterial infections in animals . Different diffusion and dilution methods, as conducted according to various approved performance standards, can be used to determine the in-vitro susceptibility of bacterial pathogens . In the present article, problems are discussed which arise from the use of different methods and the difficulty to interpret such results . While most approved performance standards were designed for testing of bacteria from human sources, the NCCLS document M31-A2 exclusively focusses on susceptibility testing of bacteria isolated from animals and--in contrast to all other standards--includes veterinary specific breakpoints for a number of antimicrobial agents used in veterinary medicine . Therefore, performance of in-vitro susceptibility testing of veterinary pathogens should follow the recommendations given in the NCCLS document M31-A2 . The microdilution method is recommended as the method of choice for susceptibility testing . The result of a microdilution test is given as the minimum inhibitory concentration (MIC) . This value provides a quantitative result which precisely indicates the degree of susceptibility of the tested bacterial strain and in return gives the veterinarian a clear guidance whether therapeutic intervention with the antibiotic in question will be successful.

J Biol Chem, 2003 Dec 19, 278(51), 51053 - 8 Epub 2003 Oct 02.
Spheniscins, avian beta-defensins in preserved stomach contents of the king penguin, Aptenodytes patagonicus; Thouzeau C et al.; During the last part of egg incubation in king penguins, the male can preserve undigested food in the stomach for several weeks . This ensures survival of the newly hatched chick, in cases where the return of the foraging female from the sea is delayed . In accordance with the characterization of stress-induced bacteria, we demonstrate the occurrence of strong antimicrobial activities in preserved stomach contents . We isolated and fully characterized two isoforms of a novel 38-residue antimicrobial peptide (AMP), spheniscin, belonging to the beta-defensin subfamily . Spheniscin concentration was found to strongly increase during the period of food storage . Using a synthetic version of one of two spheniscin isoforms, we established that this peptide has a broad activity spectrum, affecting the growth of both pathogenic bacteria and fungi . Altogether, our data suggest that spheniscins and other, not yet identified, antimicrobial substances may play a role in the long term preservation of stored food in the stomach of king penguins.

J Leukoc Biol, 2004 Jan, 75(1), 49 - 58 Epub 2003 Oct 02.
Expression and regulation of antimicrobial peptides in the gastrointestinal tract; Cunliffe RN et al.; The gastrointestinal (GI) tract is exposed to a wide range of microorganisms . The expression of antimicrobial peptides has been demonstrated in different regions of the GI tract, predominantly in epithelial cells, which represent the first host cells with which the microorganisms have to interact for invasion . The intestinal epithelial monolayer is complex, consisting of different cell types, and most have a limited lifespan . Of the GI antimicrobial peptides, alpha- and beta-defensins have been studied the most and are expressed by distinct types of epithelial cells . Enteric alpha-defensin expression is normally restricted to Paneth and intermediate cells in the small intestine . However, there are important differences between mice and humans in the processing of the precursor forms of enteric alpha-defensins . Parasite infection induces an increase in the number of enteric alpha-defensin-expressing Paneth and intermediate cells in the murine small intestine . In the chronically inflamed colonic mucosa, metaplastic Paneth cells (which are absent in the normal colon) also express enteric alpha-defensins . Epithelial expression of beta-defensins may be constitutive or inducible by infectious and inflammatory stimuli . The production of some members of the beta-defensin family appears to be restricted to distinct parts of the GI tract . Recent studies using genetically manipulated rodents have demonstrated the likely in vivo importance of enteric antimicrobial peptides in innate host defense against microorganisms . The ability of these peptides to act as chemoattractants for cells of the innate- and adaptive-immune system may also play an important role in perpetuating chronic inflammation in the GI tract.

Enferm Infecc Microbiol Clin, 2003 Oct, 21(8), 410 - 6
{Respiratory infections outside the hospital . DIRA study}; Picazo JJ et al.; INTRODUCTION: Most visits to the primary care center are for infection and particularly respiratory tract infection . Antimicrobial administration for these clinical processes is common and these agents are often used to treat viral infections . La Fundacion para el Estudio de la Infeccion (Foundation for the Study of Infection) designed the DIRA (Dia de la Infeccion Respiratoria del Adulto, Adult Respiratory Infection Day) Project to investigate and assess the impact of this problem . METHODS: The study design consisted of one-day cross sections with the participation of 720 physicians belonging to Primary Health Care Centers from the 17 Autonomic Regions of Spain, establishing a one-day cross section every three months for one year . Epidemiologic, clinical and therapeutic factors were evaluated . RESULTS: The total number of visits attended was 72 929, and 14 426 patients had infectious processes (43.9%) . Among these, 9145 (63.4%) had a respiratory infection . The mean age of the patients was 44.6 years and 34.1% had an underlying condition . Common cold was the most frequent diagnosis . Antibiotics were prescribed in 53.2% of patients . Results were compared to those of a previous study . DISCUSSION: Infection in general and respiratory infection in particular is frequently attended in primary care . Antibiotics are widely used in our setting and self-medication is common.

Pharmacotherapy, 2003 Sep, 23(9), 1190 - 4
Acute tubular necrosis associated with propylene glycol from concomitant administration of intravenous lorazepam and trimethoprim-sulfamethoxazole; Hayman M et al.; A 46-year-old morbidly obese man was admitted to the medical intensive care unit with respiratory failure . He required pressure-control ventilation and high levels of sedation with continuous-infusion lorazepam . He developed Stenotrophomonas maltophilia pneumonia; treatment included scheduled intravenous trimethoprim-sulfamethoxazole . Each of these drugs contain several hundred milligrams/milliliter of propylene glycol . On day 17 of his hospital course, 3 days after starting the trimethoprim-sulfamethoxazole, the patient developed acute renal failure consistent with acute tubular necrosis . Propylene glycol toxicity was suspected; therefore, all drugs containing propylene glycol were discontinued, and laboratory data were collected . A marked osmol gap, metabolic acidosis, and renal toxicity were attributed to both continuous and large intermittent doses of intravenous propylene glycol . Particular attention should be paid to the total amount of propylene glycol provided to patients from administered drugs . Patients in the intensive care setting who require high doses of intravenous lorazepam for sedation, as well as antimicrobial therapy with trimethoprim-sulfamethoxazole for treatment of either Stenotrophomonas maltophilia or Pneumocystis carinii pneumonia, may be at increased risk for propylene glycol toxicity and should be monitored closely.

Pharmacotherapy, 2003 Sep, 23(9), 1167 - 74
Impact of atypical coverage for patients with community-acquired pneumonia managed on the medical ward: results from the United States Community-Acquired Pneumonia Project; Frei CR et al.; STUDY OBJECTIVE: As current guidelines for treatment of community-acquired pneumonia (CAP) recommend empiric antimicrobial coverage for atypical pathogens, we evaluated the need for atypical coverage by examining length of hospital stay (LOS) and mortality in patients with CAP who were managed on the medical ward . METHODS: Medical records of patients with CAP admitted from January 1, 1997-December 31, 2001, from 176 United States nonteaching community hospitals were reviewed . Patients were divided into one of three mutually exclusive groups on the basis of intravenous antimicrobials received on days 1 or 2 of hospital stay: ceftriaxone monotherapy, ceftriaxone plus a macrolide, or levofloxacin . Variables evaluated for their ability to predict outcome were patient age, year of hospital admission, geographic region, preadmission setting, preadmission antimicrobial treatment, timing of antimicrobial administration, comorbid disease, and duration of intravenous antimicrobial treatment . The impact of initial antimicrobial regimen on LOS and mortality was evaluated in regression models while controlling for significant predictors of outcome . RESULTS: Of 8975 patients evaluated, 2453 met the inclusion criteria . Significant differences were noted among patients who received ceftriaxone (932 patients), ceftriaxone plus a macrolide (872), and levofloxacin (649) with respect to mean +/- SD age (72 +/- 16, 67 +/- 18, and 70 +/- 17 yrs, respectively; p<0.0001), admission from a nursing home (21%, 11%, and 15%, respectively; p<0.0001), and duration of intravenous antimicrobial treatment (4.4 +/- 2.7, 4.0 +/- 2.6, and 3.6 +/- 2.5 days, respectively; p<0.0001) . The LOS predictors were age, geographic region, coexisting heart failure, and duration of intravenous antimicrobial therapy . Mortality predictors were age, admission from a nursing home, coexisting heart failure, and coexisting cancer . After controlling for these predictors of outcome, no significant differences were noted among the three groups for LOS (5.5 +/- 3.5, 4.8 +/- 2.9, and 4.8 +/- 2.9 days, respectively; p=0.2791) or mortality (3.1%, 2.0%, and 2.6%, respectively; p=0.8461) . CONCLUSION: Initial coverage for atypical pathogens does not affect LOS or mortality among patients with CAP managed on the medical ward.

Pol Merkuriusz Lek, 2003 Jun, 14(84), 653 - 7
{Usefulness of antibiotics in the treatment of bronchial asthma}; Dziedziczko A; The efficacy and safety of antibiotics in the treatment of asthma are not unequivocal . Among various antibiotics, macrolides play the main role in the therapy of bronchial asthma . Macrolides are useful in the treatment of asthma not only because of their antimicrobial activity . The mechanism of action of macrolides in improving asthma and reducing airway responsiveness is evaluated especially due to their activity in non-infectious asthma . Macrolides may not only enhance the host defense system through increased cytokine synthesis, but also exhibit anti-inflammatory activity mediated by anti-inflammatory cytokines . The steroid-sparing effect of macrolide antibiotics has been postulated to contribute to their beneficial actions in the treatment of asthma . Macrolides may be useful in the treatment of patients with steroid-dependent asthma, probably because they inhibit eosinophilic inflammation . It has also been suggested that the effect of macrolides on bronchial hyperresponsiveness is mediated by their inhibitory action on superoxide production and chemotaxis of polymorphonuclear neutrophils and the mixed lymphocyte reactions . In spite of these suggestions, the mechanism of action of macrolides in asthmatic syndrome is not clear . Only well-designed and conducted clinical studies are capable of assessing the efficacy and safety of immunosuppressive macrolides in the treatment of asthma.

Cell Mol Life Sci, 2003 Sep, 60(9), 1793 - 804
Recognition of bacterial peptidoglycan by the innate immune system; Dziarski R; The innate immune system recognizes microorganisms through a series of pattern recognition receptors that are highly conserved in evolution . Peptidoglycan (PGN) is a unique and essential component of the cell wall of virtually all bacteria and is not present in eukaryotes, and thus is an excellent target for the innate immune system . Indeed, higher eukaryotes, including mammals, have several PGN recognition molecules, including CD14, Toll-like receptor 2, a family of peptidoglycan recognition proteins, Nod1 and Nod2, and PGN-lytic enzymes (lysozyme and amidases) . These molecules induce host responses to microorganisms or have direct antimicrobial effects.

J Biol Chem, 2003 Dec 19, 278(51), 51599 - 605 Epub 2003 Oct 01.
Molecular identification of Aggrus/T1alpha as a platelet aggregation-inducing factor expressed in colorectal tumors; Kato Y et al.; Platelets play an important role in hemostasis, thrombosis, and antimicrobial host defense and are also involved in the induction of inflammation, tissue repair, and tumor metastasis . We have previously characterized the platelet aggregation-inducing sialoglycoprotein (Aggrus/gp44) overexpressed on the surface of tumor cells . Because a platelet aggregation-neutralizing 8F11 monoclonal antibody that could specifically recognize Aggrus suppressed tumor-induced platelet aggregation, we have previously purified Aggrus by 8F11-affinity chromatography and found that purified Aggrus possessed the ability to induce aggregation of platelets . Here we show that Aggrus is identical to the T1alpha/gp38P/OTS-8 antigen, the function of which in tumors is unknown . Expression of mouse Aggrus and its human homologue (also known as T1alpha-2/gp36) induced platelet aggregation without requiring plasma components . Using the 8F11 antibody, we identified the highly conserved platelet aggregation-stimulating domain with putative O-glycosylated threonine residues as the critical determinant for exhibiting platelet aggregation-inducing capabilities . We compared the expression level of human aggrus mRNA using an array containing 160 cDNA pair samples derived from multiple human tumorigenic and corresponding normal tissues from individual patients . We found that expression level of aggrus was enhanced in most colorectal tumor patients . To confirm the protein expression, we generated anti-human Aggrus polyclonal antibodies . Immunohistochemical analysis revealed that Aggrus expression was frequently up-regulated in colorectal tumors . These results suggest that Aggrus/T1alpha is a newly identified, platelet aggregation-inducing factor expressed in colorectal tumors.

Diagn Microbiol Infect Dis, 2003 Oct, 47(2), 399 - 405
Prevalence of enterotoxigenic Escherichia coli (ETEC) in hospitalized acute diarrhea patients in Denpasar, Bali, Indonesia; Subekti DS et al.; The relationship between enterotoxigenic Escherichia coli (ETEC) and hospitalized patients with acute diarrhea was examined in a study conducted in two hospitals from June 2000 to May 2001 in Denpasar, Bali, Indonesia . A total of 489 hospitalized patients with acute diarrhea were enrolled, and their rectal swabs were screened for enteric bacterial pathogens . Toxins, colonization factor antigens (CFAs), in vitro antimicrobial susceptibility and seasonal distribution patterns associated with ETEC were ascertained . The diagnosis of ETEC infection and CFAs association were performed with GM-1 ELISA and Dot blot immunoassays . Enterotoxigenic Escherichia coli was isolated from the rectal swabs of 14.9% of the patients . The distribution of toxins among the ETEC strains found was ST in 51 (69.9%), while LT and ST/LT were found in 28.8% and 1.3% respectively . The highest isolation rate for ETEC was found among children between the ages of 1 and 15 years . Colonization factor antigens were identified in 28.8% of the ETEC strains . A high prevalence of CFA was found among the rectal swabs of patients with ST isolates . High frequency of resistance to ampicillin, trimethoprim/sulfamethoxazole, chloramphenicol, tetracycline and cephalothin was displayed among the ETEC strains . All ETEC strains were susceptible to norfloxacin, ciprofloxacin and nalidixic acid . The results of this study document the prevalence of ETEC in hospitalized patients with acute diarrhea in Denpasar, Bali, Indonesia . Data generated in this study depicts the prevalence of ETEC diarrhea and CFA types among diarrhea patients in the tourist city of Denpasar, Bali, Indonesia.

Ann Fr Anesth Reanim, 2003 Oct, 22(8), 711 - 5
{Risk factors for amoxicillin-clavulanate-resistant Escherichia coli in ICU patients}; Mohammedi I et al.; OBJECTIVE: To determine risk factors of infections with amoxicillin-clavulanate-resistant Escherichia coli in ICU patients . STUDY DESIGN: Prospective, consecutive sample survey study . PATIENTS: A consecutive series of 133 patients from whom culture results were positive for E . coli during their ICU stay . METHODS: Risk factors analysed included demographics, comorbid conditions, and antimicrobial drug exposure . Univariate and multivariate analysis were performed . RESULTS: Multivariate logistic regression analysis identified only one significant independent factor associated with the emergence of amoxicillin-clavulanate-resistant E . coli: prior use of amoxicillin (odds ratio: 5.45) . CONCLUSION: Clinicians should avoid administering amoxicillin-clavulanate as empiric therapy for possible E . coli infection in patients that have recently been treated with amoxicillin.

Int J Antimicrob Agents, 2003 Oct, 22(4), 429 - 38
A tissue cage model in calves for studies on pharmacokinetic/pharmacodynamic interactions of antimicrobials; Greko C et al.; An in vivo model for studies of pharmacokinetic/pharmacodynamic (PK/PD) interactions of antimicrobials was developed . Tissue cages with a constant surface area but with different volumes were implanted in calves and infected with Mannheimia haemolytica . Penicillin was injected directly into the cages . With this procedure, different concentration-time profiles could be simulated so that the effect of a range of PK/PD indices on the infection could be monitored . The area under the curve to minimum inhibitory concentration (MIC) and time above MIC were equally predictive for effect, but Cmax to MIC was not . If drug dosages in relation to the MIC of strains used for infection are optimised, the model offers an interesting alternative to explore relevant factors for drug dosage optimisation.

Int J Antimicrob Agents, 2003 Oct, 22(4), 420 - 8
National antibiotic resistance monitoring in veterinary pathogens from sick food-producing animals: the German programme and results from the 2001 pilot study; Wallmann J et al.; In 2001, the first Germany comprehensive cross-sectional study into the sensitivity to antimicrobial substances of selected pathogenic bacteria from food-producing animal species (dairy cows, fattening pigs) was conducted by the Federal Office of Consumer Protection and Food Safety, BVL (formerly, Federal Institute for Health Protection of Consumers and Veterinary Medicine, BgVV) . Initial experience from national resistance monitoring revealed that the necessary organisational structures may be suitably established in a federal system . The quantitative sensitivity results (minimum inhibitory concentration, MIC) for the bacterial species examined showed lower resistance values in contrast to German data published previously and also in comparison with results from other European countries . Based on the experience from this pilot study, an urgent need has been identified to continue this interdisciplinary approach to tackle the resistance problem together with human medicine . After analysing the data from the pilot monitoring study, in 2002 the BVL started a year's study with an extended selection of bacterial species and indications . In the future, bacterial samples from private diagnostic laboratories and universities will also be included in the resistance monitoring system.

Biotechnol Appl Biochem, 2004 Feb, 39(Pt 1), 71 - 81
The production of hypericins and hyperforin by in vitro cultures of St . John's wort (Hypericum perforatum); Kirakosyan A et al.; St . John's wort ( Hypericum perforatum L.) is a herbaceous perennial distributed throughout the World that has been widely used in traditional medicine . H . perforatum produces several types of biologically active compound, including the hypericins--a family of light-activated anthraquinones, localized within specialized glands found predominantly on flowers and leaves--and the hyperforins--a family of prenylated acylphloroglucinols localized in the reproductive structures of the plant . Hypericins are known to be toxic to mammals and display antiviral and anticancer activity, but the role of these compounds within the plant is unknown . Hyperforins display potent antimicrobial activity and are thought to be the primary bioactive ingredient for anti-depressive effects of the herb . The introduction of H . perforatum from Europe into the U.S.A . occurred in the 17th Century . Since the plant is considered a noxious weed, few efforts have been carried out to analyse populations in the context of secondary-metabolite concentrations . But in terms of secondary-metabolite studies, H . perforatum is an ideal model system to study the biosyntheses of aromatic polyketides and regulation of those pathways by environmental and genetic influences . This is due, in part, to the ease of conducting these studies in plant tissue culture . This review describes the progress of secondary-metabolite studies currently underway using H . perforatum . Specifically, this Review focuses on the production and regulation of the hypericins and the hyperforin in wild populations, field cultivation, greenhouse studies and plant tissue culture . Additionally, factors optimizing compound production--particularly in in vitro cultures--are presented and reviewed.

Expert Opin Pharmacother, 2003 Oct, 4(10), 1789 - 99
The epidemiology of severe sepsis syndrome and its treatment with recombinant human activated protein C; Doig CJ et al.; Severe sepsis syndrome has important consequences to healthcare systems as the incidence is increasing, there is significant attributed morbidity and mortality and there is a substantial cost for in-hospital and post-discharge care . Current treatment includes the use of antimicrobials, local source control and aggressive physiological support, usually in an intensive care unit setting . Drotrecogin-alpha (activated) or recombinant human activated protein C (rhAPC) is the only biological agent approved for use in severe sepsis syndrome that has demonstrated efficacy in reducing 28-day all-cause mortality and new data suggests a trend towards longer term survival . However, given the evidence of a variable effect on survival rates in patient subgroups and its acquisition cost, controversy has arisen concerning its appropriate use . This review discusses the epidemiology of sepsis, preclinical and clinical evidence supporting the use of rhAPC use, controversies about the evidence of efficacy in severe sepsis syndrome and cost-effectiveness data.

Sex Transm Dis, 2003 Oct, 30(10), 742 - 9
Spatial bridges for the importation of gonorrhea and chlamydial infection; Kerani RP et al.; A study of heterosexuals with gonorrhea and/or chlamydial infection in King County, Washington, found that 5.2% of study participants had both local and geographically distant sex partners in the 60 days before diagnosis . Individuals who served as spatial bridges were of higher socioeconomic status and older than other patients . BACKGROUND: Sexual mixing between distant geographic areas (spatial bridging) is important in the spread of antimicrobial resistance and new sexually transmitted disease pathogens . GOAL: The goal was to define the extent of sexual mixing between persons with gonorrhea or chlamydial infection in King County, Washington, and persons outside the Seattle area, and to identify characteristics of persons and partnerships associated with spatial bridging . METHODS: Patients contacted for purposes of partner notification were interviewed regarding demographics, sexual behavior, and the characteristics of their sex partners . RESULTS: Of 2912 participants, 150 (5.2%) were spatial bridgers . Bridgers were of higher socioeconomic status than nonbridgers and more often reported concurrent partnerships . Over a 39-month period, bridgers and potential bridgers linked King County with 35 states and 13 foreign countries . CONCLUSION: Spatial bridging could represent an important channel of transmission between geographic areas . These results highlight the need for linkage of prevention efforts across geographic boundaries.

J Biol Chem, 2003 Dec 5, 278(49), 48928 - 34 Epub 2003 Sep 30.
Immune activation of NF-kappaB and JNK requires Drosophila TAK1; Silverman N et al.; Stimulation of the Drosophila immune response activates NF-kappaB and JNK signaling pathways . For example, infection by Gram-negative bacteria induces the Imd signaling pathway, leading to the activation of the NF-kappaB-like transcription factor Relish and the expression of a battery of genes encoding antimicrobial peptides . Bacterial infection also activates the JNK pathway, but the role of this pathway in the immune response has not yet been established . Genetic experiments suggest that the Drosophila homolog of the mammalian MAPK kinase kinase, TAK1 (transforming growth factor beta-activated kinase 1), activates both the JNK and NF-kappaB pathways following immune stimulation . In this report, we demonstrate that Drosophila TAK1 functions as both the Drosophila IkappaB kinase-activating kinase and the JNK kinase-activating kinase . However, we found that JNK signaling is not required for antimicrobial peptide gene expression but is required for the activation of other immune inducible genes, including Punch, sulfated, and malvolio . Thus, JNK signaling appears to play an important role in the cellular immune response and the stress response.

Cleve Clin J Med, 2003 Sep, 70(9), 793 - 800
Controlling antibiotic resistance in the ICU: different bacteria, different strategies; Rice LB; To reduce antimicrobial resistance in the intensive care unit, hospitals are developing strategies such as improving infection control, adhering to prescribed formularies, requiring prior approval for using certain antibiotics, setting limits on the duration of antimicrobial therapy, and rotating the use of antimicrobial drugs on a regular schedule . Each strategy has theoretical benefits and limitations, but good data on their efficacy in controlling antimicrobial resistance are limited.

Medicina (B Aires), 2003, 63(4), 319 - 43
{Neumonía acquired in the community . Practical guide elaborated by a committee intersocieties}; Luna CM et al.; Clinical practice guidelines for community-acquired pneumonia (CAP) contribute to improve patient's management . CAP undergoes continuous changes in etiology, epidemiology and antimicrobial sensitivity, requiring periodic guidelines revisions . An inter-society committee designed this guidelines dividing it into several topics based on prior guidelines and recent clinical studies . CAP compromises annually more than 1% of the population; most of the cases only require outpatient care but others are severe cases, reaching the 6th cause of death in Argentina . The cases are distributed unevenly into ambulatory, admitted in the general ward or in the intensive care unit . There is no way to predict the etiology . Unfavorable outcome predictors include age, antecedents and physical, laboratory and radiography findings . Ten to 25% of inpatients need to be admitted to the intensive care unit at the onset or during the follow-up, for mechanical ventilation or hemodynamic support (severe CAP) . Severe CAP is associated with high mortality and requires adequate and urgent therapy . Pregnant, COPD and nursing home patients require special recommendations . Diagnosis is clinical, while complementary methods are useful to define etiology and severity; chest X-ray is the only one universally recommended . Other studies, including microbiologic evaluation are particularly appropriate in the hospitalized patients . The initial therapy is empiric, it must begin early, using antimicrobials active against the target microorganisms, avoiding their inappropriate use which can lead to the development of resistance . Length of therapy must not be unnecessarily prolonged . Hydratation, nutrition, oxygen and therapy of complications must complement antibiotic treatment . Prevention is based on influenza prophylaxis, anti-pneumococcal vaccine, aspiration prevention and other general measures.

Fungal Genet Biol, 2003 Nov, 40(2), 176 - 85
Isolation and characterization of Neurospora crassa mutants resistant to antifungal plant defensins; Ferket KK et al.; Twenty-five Neurospora crassa mutants obtained by chemical mutagenesis were screened for increased resistance to various antifungal plant defensins . Plant defensin-resistant N . crassa mutants were further tested for their cross-resistance towards other families of structurally different antimicrobial peptides . Two N . crassa mutants, termed MUT16 and MUT24, displaying resistance towards all plant defensins tested but not to structurally different antimicrobial peptides were selected for further characterization . MUT16 and MUT24 were more resistant towards plant defensin-induced membrane permeabilization as compared to the N . crassa wild-type . Based on the previously demonstrated key role of fungal sphingolipids in the mechanism of growth inhibition by plant defensins, membrane sphingolipids of MUT16 and MUT24 were analysed . Membranes of these mutants contained structurally different glucosylceramides, novel glycosylinositolphosphorylceramides, and an altered level of steryl glucosides . Evidence is provided to link these clear differences in sphingolipid profiles of N . crassa mutants with their resistance towards different plant defensins.

Biochemistry, 2003 Oct 7, 42(39), 11417 - 26
Three-dimensional structure in lipid micelles of the pediocin-like antimicrobial peptide sakacin P and a sakacin P variant that is structurally stabilized by an inserted C-terminal disulfide bridge; Uteng M et al.; The three-dimensional structures in dodecylphosphocholine (DPC) micelles and in trifluoroethanol (TFE) of the pediocin-like antimicrobial peptide sakacin P and an engineered variant of sakacin P (termed sakP{N24C+44C}) have been determined by use of nuclear magnetic resonance spectroscopy . SakP{N24C+44C} has an inserted non-native activity- and structure-stabilizing C-terminal disulfide bridge that ties the C-terminus to the middle part of the peptide . In the presence of DPC, the cationic N-terminal region (residues 1-17) of both peptides has an S-shaped conformation that is reminiscent of a three-stranded antiparallel beta-sheet and that is more pronounced when the peptide was dissolved in TFE instead of DPC . The four positively charged residues located in the N-terminal part are found pointing to the same direction . For both peptides, the N-terminal region is followed by a well-defined central amphiphilic alpha-helix (residues 18-33), and this in turn is followed by the C-terminal tail (residues 34-43 for sakacin P and 34-44 for sakP{N24C+44C}) that lacks any apparent common secondary structural motif . In the presence of DPC, the C-terminal tails in both peptides fold back onto the central alpha-helix, thereby creating a hairpin-like structure in the C-terminal halves . The lack of long-range NOEs between the beta-sheet Nu-terminal region and the hairpin-like C-terminal half indicates that there is a flexible hinge between these regions . We discuss which implications such a structural arrangement has on the interaction with the target cell membrane.

Equine Vet J, 2003 Sep, 35(6), 613 - 9
Endoscopic surgery in the treatment of contaminated and infected synovial cavities; Wright IM et al.; REASONS FOR PERFORMING STUDY: Contamination and infection of synovial cavities are a common occurrence in clinical practice and, if inadequately treated, may have career or life threatening consequences for affected horses . HYPOTHESIS: The objectives in treating contamination and infection of joints, tendon sheaths and bursae are most effectively met by endoscopic surgery . METHODS: Over a 6 year period, cases of synovial contamination and infection admitted to a referral clinic were evaluated and treated endoscopically . The horses received local and systemic antimicrobial drugs with minimal nonsteroidal anti-inflammatory medication but no other medical or surgical treatment . All arthroscope and instrument portals and, whenever possible, all traumatic wounds were closed . Diagnostic information, endoscopic observations and results of treatment were evaluated retrospectively . RESULTS: A total of 140 affected animals were referred and 121 cases were treated endoscopically . These involved 70 joints, 29 tendon sheaths, 10 bursae and in 12 cases a combination of synovial cavities . The most common aetiologies were open wounds (n = 54) and self-sealing punctures (n = 41) . Foreign material was identified endoscopically in 41 but predicted prior to surgery in only 6 cases . Osteochondral lesions were evident at surgery in 51 and recognised before surgery in 25 cases; 32 horses had intrathecal tendon or ligament defects . Follow-up information was obtained for 118 animals; 106 (90%) survived and 96 (81%) returned to their preoperative level of performance . The presence of osteitis/osteomyelitis, other osteochondral lesions and marked deposits of pannus were associated with nonsurvival . For those animals which survived, non-Thoroughbred horses, a combination of synovial structure involvement and regional i.v . antimicrobial administration were associated with reduced post operative performance . Marked pannus, regional i.v . antimicrobial administration and duration of systemic antimicrobial administration were associated with a group combining nonsurviving animals and those with reduced post operative performance . CONCLUSIONS: Endoscopic surgery makes a valuable contribution to the management of synovial contamination and infection . POTENTIAL RELEVANCE: The information obtained from and therapeutic options offered by endoscopy justify its early use in cases of synovial contamination and infection.

Acta Biochim Pol, 2003, 50(3), 743 - 52
Azurocidin -- inactive serine proteinase homolog acting as a multifunctional inflammatory mediator; Watorek W; Azurocidin, also known as cationic antimicrobial protein 37 kDa (CAP37) or heparin-binding protein (HBP) is an inactive homolog of serine proteinases residing in granulocytes . The ability to cleave peptide bond was lost due to replacement of two of the three residues from the conserved catalytic triad characteristic for serine proteinases . Azurocidin has a broad spectrum of antimicrobial activity, mainly against Gram-negative bacteria . It is also recognized as a multifunctional inflammatory mediator for its contracting effects on endothelial cells causing an increase of vascular permeability, capacity to bind endotoxin and ability to attract monocytes to inflammation sites.

Curr Opin Otolaryngol Head Neck Surg, 2003 Feb, 11(1), 1 - 5
Allergic fungal sinusitis: diagnosis and treatment; Kuhn FA et al.; Since allergic fungal sinusitis was initially described by Millar in 1981, many have tried to define and explain the disorder . It has been labeled as the sinonasal equivalent of allergic bronchopulmonary aspergillosis; however, allergic fungal sinusitis cannot be categorized so easily . According to the literature at this time, there are five major criteria and six associated characteristics or minor criteria of patients with allergic fungal sinusitis . In reality, patients may not develop all five major criteria or have any of the associated criteria for years . Allergic fungal sinusitis is not only difficult to diagnose, but it is one of the most complicated conditions rhinologists must manage . Endoscopic sinus surgery must be used in conjunction with long-term medical therapy, oral and nasal corticosteroids, immunotherapy, antifungal therapy, and antimicrobial agents to effectively control the problem . Allergic fungal sinusitis is most likely the endpoint in a spectrum of sinonasal disease, driven by the presence of fungus and eosinophils with their inflammatory mediators . The affected nasal mucosa no longer functions properly, and a cycle of chronic edema, stasis, and bacterial superinfection results . Therapy entails disrupting the inflammatory process to allow normal mucosal function to resume.

Eur J Histochem, 2003, 47(3), 215 - 22
Germ cell differentiation-dependent and stage-specific expression of LANCL1 in rodent testis; Nielsen JE et al.; LANCL1 (LanC-like protein 1) is related to the bacterial LanC (lanthionine synthetase C) family, which is involved in the biosynthesis of antimicrobial peptides . Highest expression levels of LANCL1 are found in testes and brain, two organs that exist behind blood-tissue barriers . In the mouse, the establishment of an impermeable blood-testis barrier occurs between day 10-16 post natal (pn) . Differential display analysis showed that the expression level of LANCL1 mRNA in mouse testes was very low until day 16 pn, but increased gradually from day 16 pn to reach a maximum on days 22-24 pn followed by a slight reduction from day 26 pn to adult animals . Thus, the expression of LANCL1 mRNA is initiated following the establishment of the blood-testis barrier . In situ hybridisation revealed that LANCL1 mRNA was induced in diplotene spermatocytes, which appear for the first time in mouse testes between days 18 and 20 pn, verifying the expression profile determined by differential display . LANCL1 mRNA level remained high in the meiotic division phase and in early round spermatids, but was down regulated in elongating spermatids and it was undetectable in step 9 elongating spermatids in stage IX (as defined by Russel et al., 1990) . The steady decrease in expression level from round spermatids in stage I to elongating spermatids in stage IX suggested that LANCL1 mRNA was not transcribed in spermatids . LANCL1 expression in rat testes was initiated already in pachytene spermatocytes in stage IX, but otherwise similar to mouse.

J Periodontol, 2003 Aug, 74(8), 1231 - 6
Antimicrobial periodontal treatment decreases serum C-reactive protein, tumor necrosis factor-alpha, but not adiponectin levels in patients with chronic periodontitis; Iwamoto Y et al.; BACKGROUND: Elevated levels of C-reactive protein (CRP) and decreased plasma adiponectin are associated with increased risk of atherosclerosis . Furthermore, recent observations suggested that adiponectin and tumor necrosis factor-alpha (TNF-alpha) suppressed each other's production . Since periodontal disease has been suggested to act as a risk factor for atherosclerosis, we examined the effects of antimicrobial periodontal treatment on CRP, adiponectin, and TNF-alpha levels . METHODS: Fifteen chronic periodontitis patients with various systemic conditions at high risk for atherosclerosis were enrolled in the study . Patients were non-surgically treated with topical application of antibiotics and mechanical debridement of calculus once a week for 1 month . Before and after therapy, CRP, adiponectin, and TNF-alpha levels were measured . RESULTS: Both CRP and TNF-alpha levels were significantly decreased after treatment (P<0.01 and P<0.03, respectively), while adiponectin levels did not change significantly . CONCLUSIONS: Periodontal treatment is effective in reducing CRP and TNF-alpha, while adiponectin does not appear to be influenced by periodontal treatment . Elevated levels of CRP and TNF-alpha may be associated with increased risk for future development of atherosclerosis in periodontitis patients.

J Periodontol, 2003 Aug, 74(8), 1191 - 5
Effects of enamel matrix derivative on Porphyromonas gingivalis; Newman SA et al.; BACKGROUND: Enamel matrix derivative (EMD) is used during periodontal surgery for the regeneration of periodontal tissue . It consists of the amelogenin fraction of porcine enamel matrix (AMEL) suspended in a vehicle of propylene glycol alginate (PGA) . EMD-treated sites appear to heal with less inflammation . It has been suggested that antimicrobial properties of EMD might account for improved healing in vivo . The objectives of this study were: 1) to determine the antibacterial effects of EMD on the periodontal pathogen Porphyromonas gingivalis and 2) to establish the component(s) of EMD that are responsible for this effect . METHODS: The antimicrobial effects were determined in vitro using the broth dilution assay . P . gingivalis at a starting inoculum of 10(9) colony forming units/ml was treated with EMD, AMEL, and PGA in Hank's balanced salt solution (HBSS) for 1, 3, and 24 hours . The CFU/ml of P . gingivalis recovered on enriched Brucella blood agar was determined at 6 and 10 days . RESULTS: EMD (containing AMEL and PGA) or PGA alone eliminated recoverable CFUs of P . gingivalis . Interestingly, AMEL in HBSS increased recoverable CFUs from 8.62 log CFU/ml to 8.93 log CFU/ml . Further analysis of the dose response at concentrations of 0.3, 3, and 30 mg/ml of AMEL in HBSS revealed that only 30 mg/ml (clinical concentration) increased CFUs of P . gingivalis relative to baseline (from 8.8 log CFU/ml to 9.2 log CFU/ml in 3 hours) . Additionally, AMEL was compared to a protein control bovine serum albumin (BSA) to determine whether this effect was unique to AMEL . A marked increase in recoverable CFUs occurred with the AMEL (increasing from 8.8 log CFU/ml to 9.47 log CFU/ml), but not with BSA . CONCLUSIONS: EMD possesses antimicrobial properties that can be attributed to the propylene glycol alginate vehicle . The amelogenin fraction of porcine enamel matrix in enamel matrix derivative (i.e., AMEL) is not antibacterial for P . gingivalis and was shown to increase recoverable CFUs.

Biotechnol Lett, 2003 Aug, 25(16), 1317 - 23
An antimicrobial peptide of the earthworm Pheretima tschiliensis: cDNA cloning, expression and immunolocalization; Wang X et al.; A cDNA encoding a putative antimicrobial peptide (named PP-1) was obtained using a rapid amplification of cDNA ends from the Asian earthworm, Pheretima tschiliensis . PP-1 showed 77.6% homology with the antimicrobial peptide lumbricin I isolated from the earthworm Lumbricus rubellus . PP-1 lacked an obvious signal peptide sequence . RT-PCR analysis demonstrated that this gene was expressed mainly in the body wall . PP-1 was expressed in Escherichia coli as a fusion protein with a maltoze-binding protein . A polyclonal antiserum was raised in mice using this recombinant fusion protein as antigen . Immunohistochemical studies showed that PP-1 was only in the mucus of the epidermis.

Biotechnol Lett, 2003 Aug, 25(16), 1305 - 10
Antibiotic activity of Leu-Lys rich model peptides; Park Y et al.; To develop novel antibiotic peptides useful as therapeutic drugs, short model peptides rich in Leu and Lys were designed by changing not only the net positive charge by Lys-deletion but also in the hydrophobic helix region by Leu-deletion from a peptide analogue of cecropin A (1-8)-magainin 2 (1-12) (CA-MA) known as P5 . In particular, one peptide (P6), which was obtained by deleting Lys residues (positions 1, 3, 5, 9, 10, 13, 14) and Leu residues (positions 4, 7, 8, 11, 12, 15) and keeping Pro (position 6) and Trp (position 2), showed a strong antimicrobial and antitumor activity at 0.2-3.1 microM without hemolytic activity against human erythrocyte cells . Furthermore, P6 causes significant morphological alterations of the bacterial surfaces at 3.1 microM as shown by scanning electron microscopy.

J Arthroplasty, 2003 Sep, 18(6), 714 - 8
Efficacy of a single dose of cefazolin as a prophylactic antibiotic in primary arthroplasty; Tang WM et al.; We analyzed the wound infection rate of 1,367 primary total hip and knee arthroplasties performed between 1991 and 1999 . Two hundred and fifteen arthroplasties were performed with 3 doses (3 x 750 mg) of cefuroxime, and 1,152 arthroplasties were performed with a single preoperative dose (1 x 1 g) of cefazolin as antimicrobial prophylaxis . All wound infections that occurred within 2 years of the index surgery were analyzed . The deep wound infection rate of total hip arthroplasty was 1.1% (95% confidence interval {CI}, 0%-3.3%) in the cefuroxime group and 1.1% (95% CI, 0%-2.2%) in the cefazolin group (Fisher's exact test, P = 1.0) . The deep wound infection rate of total knee arthroplasty in the cefuroxime group (1.6%; 95% CI, 0%-3.8%) was not significantly different from the cefazolin group (1.0%; 95% CI, 0.3%-1.7%) (Fisher's exact test, P =.63) . We concluded that a single dose (1 g) of cefazolin given at anesthetic induction offered similar protection to 3 doses (3 x 750 mg) of cefuroxime in preventing infection in primary total joint arthroplasty.

J Infect Chemother, 2003 Sep, 9(3), 276 - 7
Pharmacokinetics of single-dose intravenous ciprofloxacin in blood and ascites of patients with pelvic peritonitis; Mikamo H et al.; To estimate the efficacy of ciprofloxacin in the treatment of pelvic peritonitis, a pharmacokinetic study was conducted in four Japanese subjects . Ciprofloxacin was administered intravenously at a dose of 300 mg for 1 h to patients with pelvic peritonitis . Ascites was collected by culdocentesis . The concentrations of ciprofloxacin in blood and ascites were measured by a bioassay, using Escherichia coli Kp as the test organism and heart infusion agar as the medium . The ciprofloxacin concentration in ascites ranged from 3.01 to 9.41 microg/ml . The values for the arithmetic mean of ascites/serum ranged from 4.01 to 19.37, which showed that the penetration of ciprofloxacin was higher in ascites than in serum . The concentrations of ciprofloxacin are generally higher in ascites than in blood . Taking the antimicrobial spectrum of ciprofloxacin and the causative organisms of pelvic peritonitis into account, intravenous ciprofloxacin could be a good candidate for use in the treatment of pelvic peritonitis from the point of view of pharmacokinetics.

J Infect Chemother, 2003 Sep, 9(3), 268 - 71
Successful treatment of pyoderma gangrenosum that developed in a patient with myelodysplastic syndrome; Yamauchi T et al.; We describe the successful treatment of pyoderma gangrenosum (PG) that developed in a patient with myelodysplastic syndrome (MDS) . A 63-year-old Japanese man with MDS was admitted to our hospital because of a large skin ulcer on his neck in November 2001 . The initial diagnosis was infectious dermatitis, and antimicrobial therapy was performed, using imipenem/cilastatin, isepamicin, and amphotericin B . However, this therapy was not effective, and the lesion worsened . Cultures of blood, throat swab, and ulcer pus yielded no microorganisms . A biopsy of the skin lesion revealed a severe infiltration of neutrophils in the dermis, without any evidence of infection . The lesion was finally diagnosed as PG, and systemic administration of corticosteroid hormone was started in December 2001 . The patient was initially pulsed with 1 g methylprednisolone daily for 3 days . The dose was immediately reduced, and the treatment was maintained with 30 mg prednisolone daily . The skin lesion responded markedly to the therapy, and C-reactive protein became negative . The patient was discharged in February 2002 because the lesion was almost cured . Prednisolone administration was tapered after 6-month maintenance therapy . No recurrence of PG was seen, although his MDS transformed into leukemia in April 2003 . Only 31 cases of MDS developing PG have been reported in the past 20 years in Japan . This report describes one such rare patient who was successfully treated with the use of high-dose pulse methylprednisolone and long-term maintenance therapy.

Hautarzt, 2003 Oct, 54(10), 937 - 45
{Systemic therapy of atopic dermatitis}; Heratizadeh A et al.; The optimal treatment of atopic dermatitis requires regular medical supervision . The course of this chronic skin disease is influenced by multiple triggers which are relevant for the treatment . The mainstays of topical therapy include regular use of emollients coupled with antimicrobial substances, corticosteroids and immune modulators as required . Ultraviolet radiation and immunosuppressive regimens represent further options for the treatment of severe exacerbations and may lead to long term improvement . Data from experimental studies provide insight into possible future treatment methods.

Br J Dermatol, 2003 Sep, 149(3), 590 - 7
Tolerability and efficacy of N-chlorotaurine in comparison with chloramine T for the treatment of chronic leg ulcers with a purulent coating: a randomized phase II study; Nagl M et al.; BACKGROUND: The well-known active chlorine compound chloramine T (CAT) with broad-spectrum antimicrobial activity is in common therapeutic use for leg ulcers with purulent coatings; however, this treatment is painful . The tolerability of the less aggressive N-chlorotaurine (NCT), an endogenous compound also produced in vivo by stimulated human granulocytes, could be superior . OBJECTIVES: To assess the tolerability and efficacy of NCT in the cleaning of purulent coatings in chronic leg ulcers in comparison with CAT . METHODS: In a double-blind, randomized phase IIb clinical study 40 patients were treated for a median of 7 days (range 3-14) with a 1% aqueous solution of either NCT (20 subjects) or CAT (20 subjects) by twice-daily application of dressings soaked in the test solutions . Criteria for evaluation of tolerability were intensity and duration of pain caused by the ulcer therapy and scores of tissue toxicity (necrosis, granulation tissue and re-epithelialization) . Therapeutic efficacy was graded as scores of intensity of purulent coating of the ulcers . RESULTS: The concentration tolerated in vitro by human epidermoid carcinoma cells was at least 10-fold higher for NCT (0.01%) compared with CAT (0.0001-0.001%) . There was significantly less pain caused by NCT compared with CAT (P < 0.05) on days 1 and 4 and a trend for a shorter duration of pain (P = 0.093) . The scores of intensity of coating improved without difference in both treatment groups, whereas granulation and re-epithelialization appeared earlier in the NCT group (P < 0.05) . Non-quantitative microbiological cultures from ulcer smears revealed persistence of colonization by bacterial species in approximately half of both treatment groups . CONCLUSIONS: Both active chlorine compounds were helpful in reducing purulent coatings . Because of its lower toxicity and better tolerability, NCT is of advantage in the treatment of leg ulcers.

Br J Dermatol, 2003 Sep, 149(3), 484 - 91
Proinflammatory cytokines upregulate expression of calprotectin (L1 protein, MRP-8/MRP-14) in cultured human keratinocytes; Mork G et al.; BACKGROUND: Normal skin contains no epidermal calprotectin . In biopsies from various inflammatory skin disorders, however, this antimicrobial protein occurs in the cytoplasm of keratinocytes . OBJECTIVES: To exclude the possibility of epidermal uptake of calprotectin from granulocytes and macrophages in diseased skin, we investigated whether cytokine-stimulated human keratinocytes can express calprotectin in vitro . METHODS: Keratinocytes from healthy individuals were cultured in serum-free keratinocyte medium . The cells were stimulated with different cytokines {interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-10 and IL-13}, both separately and in various combinations . Cytoplasmic protein levels of calprotectin were measured by an enzyme-linked immunosorbent assay performed on fixed adherent keratinocytes, and mRNA expression was determined by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) . RESULTS: Calprotectin was produced by cytokine-stimulated keratinocytes, especially in response to combinations of the proinflammatory cytokines, which showed an additive upregulatory effect . When expression of mRNA for the light (MRP-8) and heavy (MRP-14) calprotectin chain was determined by RT-PCR, their respective levels were shown to be increased four- to ninefold and three- to fivefold after 24 h of combined stimulation with IFN-gamma and TNF-alpha . The time course of calprotectin production showed no significant elevation for the first 16 h but then increased and peaked after 36 h . CONCLUSIONS: Cultured human keratinocytes stimulated with proinflammatory cytokines produce calprotectin, suggesting that epidermal expression of this antimicrobial protein in diseased skin reflects compartmentalized synthesis rather than uptake from dermal inflammatory cells.

Acta Neurol Scand Suppl, 2003, 180, 16 - 22
Epilepsy and comorbidity: infections and antimicrobials usage in relation to epilepsy management; Sander JW et al.; Infections are probably the most common preventable cause of epilepsy worldwide . There are concerns that endemic infections and infestations, such as malaria and neurocysticercosis, could be responsible for the increased incidence of epilepsy in the developing world . Cases of epilepsy associated with neurocysticercosis are also being seen increasingly in developed countries due to migration from, and travel to, endemic areas . When prescribing antimicrobial agents in patients with epilepsy a number of issues need to be considered, such as potential adverse effects on seizure control and interactions with concomitant antiepileptic drugs (AEDs) . Some antimicrobial agents, including penicillins, cephalosporins, carbapenems, quinolones and antimalarials, can have proconvulsant activity and may precipitate seizures, even in patients who do not have epilepsy . Moreover, many antimicrobials increase or decrease the plasma levels of AEDs, whereas some AEDs may adversely affect the efficacy of antimicrobials.

J Nat Prod, 2003 Sep, 66(9), 1291 - 3
Isolation and structure determination of an antimicrobial ester from a marine sediment-derived bacterium; Schumacher RW et al.; A new compound, assigned the trivial name bonactin (1), has been isolated from the liquid culture of a Streptomyces sp . BD21-2 obtained from a shallow-water sediment sample collected at Kailua Beach, Oahu, Hawaii . Structure elucidation employed one- and two-dimensional NMR, HRFABMS, IR, and chemical analysis . Bonactin displayed antimicrobial activity against both Gram-positive and Gram-negative bacteria as well as antifungal activity.

J Nat Prod, 2003 Sep, 66(9), 1242 - 4
Divinatorins A-C, new neoclerodane diterpenoids from the controlled sage Salvia divinorum; Bigham AK et al.; Three new neoclerodane diterpenoids, divinatorins A-C (7-9), have been isolated from the leaves of Salvia divinorum . The compounds were identified by spectroscopic methods as derivatives of the antibiotic (-)-hardwickiic acid (10), which was also isolated, along with four other known terpenoids . Neither the crude extract nor 7-9 displayed antimicrobial activity.

J Nat Prod, 2003 Sep, 66(9), 1149 - 53
Purification, structure determination, and antimicrobial activity of neutramycins B-G; Graziani EI et al.; Neutramycins B-G were purified from a historical sample of neutramycin in our antibiotic collection . The structures of the compounds were solved by 2D NMR spectroscopic analysis . Four of the compounds (2-5) are probable biosynthetic intermediates or shunt metabolites of neutramycin biosynthesis, while two (6, 7) are likely to be degradation products . Only one intermediate (5) showed weak Gram-positive activity.

Infect Control Hosp Epidemiol, 2003 Sep, 24(9), 662 - 6
Duration of empiric antibiotics for suspected early-onset sepsis in extremely low birth weight infants; Cordero L et al.; OBJECTIVES: To study multicenter antibiotic practices for suspected early-onset sepsis (EOS) with negative blood cultures (NegBCs) and to identify opportunities for reduction of antimicrobial exposure . DESIGN: Retrospective study . SETTING: Thirty academic hospitals (University HealthSystem Consortium) located in 24 states . METHODS: Data were from a survey of 790 extremely low birth weight (ELBW) infants . Total antibiotic exposures (antibiotic-days per patient) were calculated . RESULTS: On admission to the NICU, 94% of 790 ELBW infants had BCs performed and empiric antibiotics initiated . When PosBC and NegBC infants were compared, 47 patients with PosBCs were similar to 695 with NegBCs in birth weight, gestational age (GA), and mortality . Patients with suspected EOS but NegBCs given ampicillin/aminoglycosides were grouped by length of administration and GA . For GA of 26 weeks or younger, 170 infants given a short (< or = 3 days) and 157 given a long (> or = 7 days) course were similar regarding birth weight, mortality, antepartum history, and CRIB scores, but were different (P < .01) in number receiving a third antimicrobial (3% and 17%) and antibiotic-days (23 and 38) . For GA of 27 weeks or older, 113 infants given a short and 77 given a long course differed (P < .01) in number receiving a third antimicrobial (2% and 23%) and antibiotic-days (19 and 30) . CONCLUSIONS: Most suspected EOS infants with NegBCs are given antibiotics, but no antepartum historical risk factors or neonatal clinical signs explained prolonged administration . Discontinuing empiric antibiotics when BCs are negative in asymptomatic ELBW infants can reduce antimicrobial exposure without compromising clinical outcome.

Vet Res Commun, 2003 Jul, 27(5), 341 - 57
Characterization of Escherichia coli isolates incriminated in colisepticaemia in mink; Tibbetts RJ et al.; Colisepticaemia is a major health and economic concern for the mink industry, yet little information is available about the Escherichia coli that cause this disease . In this study, 40 E . coli, isolated from mink clinically diagnosed with colisepticaemia that had been submitted to the North Dakota State University Veterinary Diagnostic Laboratory, were randomly selected for characterization . These isolates were serotyped and screened for resistance to 18 antimicrobials, possession of transmissible R plasmids, and the presence of several virulence traits or genes using bioassays or the polymerase chain reaction . Several serotypes were identified that have previously been associated with septicaemia in other animal species . The majority of the isolates exhibited multiple antimicrobial resistance phenotypes . Common resistance phenotypes observed included those to tetracycline, sulfamethoxazole, streptomycin, ampicillin and kanamycin . Several of the isolates that could be studied by conjugation contained transmissible R plasmids coding for multiple antimicrobial resistance phenotypes . About half of the isolates produced colicin; all produced enterobactin: and all but one-quarter produced aerobactin . None of the isolates tested produced enterohaemolysin, and one-fifth were considered to be beta haemolytic . About half appeared to contain the gene encoding cytotoxic necrotizing factor-1; three contained the gene encoding EAE, but none appeared to contain the genes coding for LT, Sta/b, SLT-I/II or CNF-II toxins or K99 antigen . Approximately one-third of the isolates elaborated capsule . The results show that the E . coli isolates implicated in mink colisepticaemia possess similar virulence traits and antimicrobial resistance phenotypes to those associated with diarrhoeal diseases in food animals.

Curr Opin Investig Drugs, 2003 Aug, 4(8), 991 - 8
Pharmacokinetics and pharmacodynamics in the development of antifungal compounds; Andes D; Antimicrobial pharmacodynamics describe the relationship between drug exposure and treatment outcome . Pharmacodynamic studies provide information useful for dose level and dosing interval selection and for the development of in vitro susceptibility guidelines . Pharmacodynamic observations from animal model studies have proven useful for outcome predictions in the treatment of human infections . The strength of these predictions has been demonstrated most clearly for antibacterial drugs . Recent studies suggest that similar analyses may also be useful for antifungal drug development.

Curr Opin Investig Drugs, 2003 Aug, 4(8), 944 - 52
Gram-positive bacterial resistance: future treatment options; Bassetti M et al.; Gram-positive infections are a major burden on patients and healthcare systems globally, and the need to treat these infections correctly in an empirical manner has become paramount . Further complicating this changing etiology is the emergence of resistant strains which are no longer predictably susceptible to standard first-line antimicrobials such as oxacillin or vancomycin . Thus, new agents such as linezolid have been developed to alleviate the 'guesswork' of initial empirical prescribing in infections where Gram-positive pathogens may be present . Future agents also being developed for multiresistant Gram-positive infections include evernimicin antibiotics, daptomycin, oritavancin, glycylcyclines and novel broad-spectrum cephalosporins; however, these are still in the development phase.

Curr Opin Investig Drugs, 2003 Aug, 4(8), 937 - 43
Bacterial RNase P as a potential target for novel anti-infectives; Eder PS et al.; The diversity of higher-order structure in ribonucleoprotein (RNP) complexes makes them amenable to small-molecule modulation of their biological function . This review will discuss why bacterial RNase P, a simple yet essential ubiquitous and highly conserved RNP enzyme, represents an excellent target for the discovery and development of novel antimicrobials.

J Cardiovasc Pharmacol, 2003 Oct, 42(4), 469 - 76
Minocycline inhibits smooth muscle cell proliferation, migration and neointima formation after arterial injury; Pinney SP et al.; The tetracyclines are antimicrobials that also inhibit expression of certain matrix metalloproteinases (MMPs) . We conducted a series of experiments to determine if minocycline could inhibit MMP expression and limit human aortic smooth muscle cell (SMC) proliferation and migration . Analysis of SMC proliferation was performed after cells were grown in minocycline-incubated media . SMC migration activity was assayed in a micro-Boyden chamber . Western blotting revealed that minocycline reduced SMC production of MMP-2 in a dose dependent manner . Increasing doses of minocycline progressively reduced SMC proliferation to 49% of control values and limited SMC migration to 15% of control . When administered to rats with balloon injured carotid arteries, intraperitoneal doses of minocycline (70-100 mg/kg) reduced neointima formation by 76%, but were associated with liver toxicity . Higher doses were lethal and lower doses were ineffective . Minocycline, applied to injured arteries in a pluronic gel with a low pH, was also ineffective . In summary, minocycline lowers MMP-2 expression, reduces SMC proliferation and migration, and inhibits neointimal hyperplasia, but its efficacy is limited by systemic toxicity.

Plant Cell, 2003 Nov, 15(11), 2626 - 35 Epub 2003 Sep 24.
A tale of three cell types: alkaloid biosynthesis is localized to sieve elements in opium poppy; Bird DA et al.; Opium poppy produces a diverse array of pharmaceutical alkaloids, including the narcotic analgesics morphine and codeine . The benzylisoquinoline alkaloids of opium poppy accumulate in the cytoplasm, or latex, of specialized laticifers that accompany vascular tissues throughout the plant . However, immunofluorescence labeling using affinity-purified antibodies showed that three key enzymes, (S)-N-methylcoclaurine 3'-hydroxylase (CYP80B1), berberine bridge enzyme (BBE), and codeinone reductase (COR), involved in the biosynthesis of morphine and the related antimicrobial alkaloid sanguinarine, are restricted to the parietal region of sieve elements adjacent or proximal to laticifers . The localization of laticifers was demonstrated using antibodies specific to the major latex protein (MLP), which is characteristic of the cell type . In situ hybridization showed that CYP80B1, BBE, and COR gene transcripts were found in the companion cell paired with each sieve element, whereas MLP transcripts were restricted to laticifers . The biosynthesis and accumulation of alkaloids in opium poppy involves cell types not implicated previously in plant secondary metabolism and dramatically extends the function of sieve elements beyond the transport of solutes and information macromolecules in plants.

Invest Ophthalmol Vis Sci, 2003 Oct, 44(10), 4412 - 8
Effect of human cationic antimicrobial protein 18 Peptide on endotoxin-induced uveitis in rats; Ohgami K et al.; PURPOSE . Human cationic antimicrobial protein 18 (hCAP18, 18 kDa) was originally identified in leukocytes on the basis of its antimicrobial activity . The peptide composed of the 27 C-terminal amino acids of hCAP18 (hCAP18(109-135)) binds lipopolysaccharide (LPS) . The purpose of the present study was to investigate the effects of hCAP18 peptide on endotoxin-induced uveitis (EIU) in rats . METHODS . EIU was induced by footpad injection of LPS . Each rat was injected intravenously with 1, 10, or 100 micro g hCAP18 peptide in 0.1 mL of PBS immediately after LPS injection in male Lewis rats . At 24 hours after LPS injection, enzyme-linked immunosorbent assay was performed to evaluate concentrations of protein, nitric oxide (NO), tumor necrosis factor (TNF)-alpha, prostaglandin (PG)-E2, interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1 and macrophage inflammatory protein (MIP)-2 in aqueous humor . Also, EIU was evaluated by counting inflammatory cells in aqueous humor . RESULTS . hCAP18 peptide at 10 and 100 micro g significantly suppressed an LPS-induced increase in the number of inflammatory cells and the levels of protein, NO, TNF-alpha, PGE2, MCP-1, and MIP-2 . The anti-inflammatory effect of 10 micro g hCAP18 peptide was as strong as that of 100 micro g hCAP18 peptide . Treatment with 1 micro g hCAP18 peptide did not suppress EIU, compared with the LPS group . CONCLUSIONS . The present results indicate that hCAP18 peptide suppresses development of EIU . A possible mechanism for the ocular anti-inflammatory effect of hCAP18 peptide is that it suppresses onset of LPS-triggered inflammatory reactions by binding directly to LPS.

Biophys J, 2003 Oct, 85(4), 2650 - 60
Aggregation of puroindoline in phospholipid monolayers spread at the air-liquid interface; Dubreil L et al.; Puroindolines, cationic and cystine-rich low molecular weight lipid binding proteins from wheat seeds, display unique foaming properties and antimicrobial activity . To unravel the mechanism involved in these properties, the interaction of puroindoline-a (PIN-a) with dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) monolayers was studied by coupling Langmuir-Blodgett and imaging techniques . Compression isotherms of PIN-a/phospholipid monolayers and adsorption of PIN-a to lipid monolayers showed that the protein interacted strongly with phospholipids, especially with the anionic DPPG . The electrostatic contribution led to the formation of a highly stable lipoprotein monolayer . Confocal laser scanning microscopy and atomic force microscopy showed that PIN-a was mainly inserted in the liquid-expanded phase of the DPPC, where it formed an aggregated protein network and induced the fusion of liquid-condensed domains . For DPPG, the protein partitioned in both the liquid-expanded and liquid-condensed phases, where it was aggregated . The extent of protein aggregation was related both to the physical state of phospholipids, i.e., condensed or expanded, and to the electrostatic interactions between lipids and PIN-a . Aggregation of PIN-a at air-liquid and lipid interfaces could account for the biological and technological properties of this wheat lipid binding protein.

Biophys J, 2003 Oct, 85(4), 2363 - 73
Solid-state NMR investigation of the depth of insertion of protegrin-1 in lipid bilayers using paramagnetic Mn2+; Buffy JJ et al.; The depth of insertion of an antimicrobial peptide, protegrin-1 (PG-1), in lipid bilayers is investigated using solid-state NMR . Paramagnetic Mn(2+) ions bind to the surface of lipid bilayers and induce distance-dependent dipolar relaxation of nuclear spins . By comparing the signal dephasing of the peptide with that of the lipids, whose segmental depths of insertion are known, we determined the depths of several residues of PG-1 in 1,2 dilauryl-sn-glycero-3-phosphotidylcholine (DLPC) bilayers . We found that residues G2 at the N-terminus and F12 at the beta-turn of the peptide reside near the membrane surface, whereas L5 and V16 are embedded in the acyl chain region . The depths increase in the order of G2 < F12 < L5 < V16 . These intensity-dephasing results are confirmed by direct measurement of the paramagnetically enhanced (13)C transverse relaxation rates . The relative depths indicate that PG-1 is tilted from the bilayer normal, which is consistent with independent solid-state NMR measurements of PG-1 orientation in the same lipids (Yamaguchi et al., 2001) . They also indicate that PG-1 is fully immersed in the lipid bilayer . However, a quantitative mismatch between the bilayer thickness and PG-1 length suggests a local thinning of the DLPC bilayer by 8-10 A . The depth sensitivity of this Mn(2+) dephasing technique is tunable with the Mn(2+) concentration to focus on different regions of the lipid bilayer.

J Am Vet Med Assoc, 2003 Sep 15, 223(6), 846 - 51
Evaluation of effect of estradiol cypionate administered prophylactically to postparturient dairy cows at high risk for metritis; Overton MW et al.; OBJECTIVE: To determine whether 4 mg of estradiol cypionate (ECP) administered prophylactically to high-risk postparturient dairy cows decreases incidence of postpartum metritis . DESIGN: Randomized, placebo-controlled, triple-masked clinical trial . ANIMALS: 250 postparturient dairy cows in a herd with postparturient hypocalcemia, retained fetal membranes, dystocia, stillbirth, or twins . PROCEDURE: Cows were given 4 mg of ECP (treatment) or 2 mL of vegetable oil (control) by i.m . injection within 24 to 36 hours of calving . Monitoring rectal temperatures and evaluation for metritis was performed once daily for 10 days . Cows with fever > or = 39.7 degrees C (103.5 degrees F) were treated with 1.5 g of ceftiofur hydrochloride . RESULTS: When assessed by ordinal logistic regression, there were no differences between groups in incidence of mild or severe metritis . Cows that calved during the second or third quarter of the year were at increased risk of metritis, compared with those that calved during the fourth quarter . Following stratification by lactation (first and > or = 2), it was observed that multiparous cows that did not receive antimicrobials during the first 3 days of the postparturient period were 5 times as likely to have metritis, compared with cows treated with antimicrobials on the basis of fever or other concurrent disease . CONCLUSIONS AND CLINICAL RELEVANCE: Prophylactic administration of ECP to dairy cows at high risk for metritis did not reduce risk for metritis . Treating multiparous cows with antimicrobials on the basis of fever during the early postpartum period was associated with decreased incidence of metritis.

Vet Ther, 2003 Summer, 4(2), 110 - 9
Characterization of antimicrobial aerosols for administration to horses; McKenzie HC; A study was conducted to characterize the aerosols produced by a medical ultrasonic nebulizer using solutions containing antimicrobials appropriate for therapy of equine lower respiratory bacterial infections (gentamicin sulfate and ceftiofur sodium) . Test aerosols were generated using an ultrasonic nebulizer and were analyzed using a laser diffraction aerosol particle analyzer . The aerosol was described in terms of the particle size distribution (volume median diameter), span (sample dispersion), and aerosol density (% volume) . The particle size distribution and aerosol density of gentamicin and ceftiofur aerosols were affected by the antimicrobial concentration of the solution . All solutions produced aerosols appropriate for delivery of antimicrobials to the intrathoracic airways of the horse, but the gentamicin (50 mg/ml) and ceftiofur (25 mg/ml) solutions offered the optimal combinations of particle size and aerosol density.

J Pediatr (Rio J), 2003 May-Jun, 79(3), 209 - 14
{Invasive pneumococcal strains isolated from children and adolescents in Salvador}; Nascimento-Carvalho CN et al.; OBJECTIVES: To describe the antimicrobial resistance and serotype distribution of pneumococcal strains . METHODS: In a 57-month period, a laboratory-based surveillance of invasive pneumococcal strains from patients aged < 20 years was conducted . Pneumococcus was identified by means of tests for solubility in bile and optochin . Pneumococcal resistance to penicillin was screened by 1 micro g oxacillin disc and minimal inhibitory concentration was determined for the strains not susceptible to penicillin . Disc diffusion and broth microdilution methods were used for surveillance of resistance to other antimicrobials . Pneumococci were serotyped by means of the Neufeld-Quellung reactions . RESULTS: Of 70 patients, 57.1% were males . The mean age was 1.92 yrs (mean 3.19 +/- 3.66 yrs, range 1 month to 19.5 yrs); 52.9% and 81.4% were < 2 yrs and < 5 yrs, respectively . The strains were isolated from blood (91.4%), CSF (2.9%), pleural (2.9%), peritoneal (1.4%) and abscess (1.4%) fluids from patients with pneumonia (77.1%), fever without localizing signs (10.0%), meningitis (4.3%), others (8.6%) . Resistance was detected to penicillin (20.0%), trimethoprim-sulfamethoxazole (65.7%), tetracycline (21.4%), ofloxacin (6.3%), erythromycin (5.7%), clindamycin (2.9%) . All tested strains were susceptible to chloramphenicol and vancomycin . Among penicillin-resistant strains, high resistance was detected in one, the same that showed intermediate resistance to cefotaxime . The most frequent serotypes were: 14 (22.9%), 5 and 6A (10.0% each), 6B and 19F (8.6% each), 9V, 18C and 23F (5.7% each) . Resistance to penicillin was detected in serotypes 14 (71.4%), 6B and 19F (14.3% each) . CONCLUSIONS: Of 70 strains, 67.2% were classified as serotypes included in the heptavalent conjugate pneumococcal vaccine, as were all penicillin-resistant strains.

Sci STKE . 2003 Sep 23;2003(201):PE39.
OutFOXing the grim reaper: novel mechanisms regulating longevity by forkhead transcription factors; Coffer P; Studies in the nematode Caenhorhabditis elegans have contributed to understanding the mechanisms that control life-span and aging . Recent experiments have employed RNAi to knock down gene expression, microarray analysis to identify genes involved in the insulin/insulin-like growth factor pathway, and bioinformatics screening to identify new Forkhead-regulated genes . The results from these experiments suggest that members of the Forkhead transcription factor family along with heat shock factors may coordinately regulate production of small heat shock proteins to control how cell stress influences life-span, and provide new links between metabolism and longevity, as well as indicate that antimicrobial genes may be involved in the aging process.

J Exp Biol, 2003 Nov, 206(Pt 21), 3835 - 43
Transgenesis and reverse genetics of mosquito innate immunity; Shin SW et al.; In recent years, mosquito molecular biology has been a scene of astounding achievements, namely the development of genetic transformation, characterization of inducible tissue-specific promoters, and acquirement of mosquito genome sequences . However, the lack of a complete genetic tool box for mosquitoes remains a serious obstacle in our ability to study essential mosquito-specific mechanisms . Unlike Drosophila, very few null mutations for mosquito genes exist . The development of reverse-genetic analyses based on RNAi and transgenic techniques will help to compensate for these deficiencies and aid in identification of critical genes in important regulatory pathways . The study of mosquito innate immunity is one example and described here . In this study, we combine mosquito transgenesis with reverse genetics . The advantage of transgenesis is the ability to establish genetically stable, dominant-negative and overexpression phenotypes . Using the blood-meal-activated vitellogenin gene (Vg) promoter, we have generated transgenic mosquitoes with blood-meal-activated, overexpressed antimicrobial peptides, Defensin A and Cecropin A . Moreover, we have recently generated a transgenic dominant-negative Relish mosquito strain, which after taking a blood meal, becomes immune-deficient to infection by Gram-negative bacteria . The latter accomplishment has opened the door to a reverse-genetic approach in mosquitoes based on transgenesis.

Antimicrob Agents Chemother, 2003 Oct, 47(10), 3290 - 5
Identifying antimicrobial resistance genes with DNA microarrays; Call DR et al.; We developed and tested a glass-based microarray suitable for detecting multiple tetracycline (tet) resistance genes . Microarray probes for 17 tet genes, the beta-lactamase bla(TEM-1) gene, and a 16S ribosomal DNA gene (Escherichia coli) were generated from known controls by PCR . The resulting products (ca . 550 bp) were applied as spots onto epoxy-silane-derivatized, Teflon-masked slides by using a robotic spotter . DNA was extracted from test strains, biotinylated, hybridized overnight to individual microarrays at 65 degrees C, and detected with Tyramide Signal Amplification, Alexa Fluor 546, and a microarray scanner . Using a detection threshold of 3x the standard deviation, we correctly identified tet genes carried by 39 test strains . Nine additional strains were not known to harbor any genes represented on the microarray, and these strains were negative for all 17 tet probes as expected . We verified that R741a, which was originally thought to carry a novel tet gene, tet(I), actually harbored a tet(G) gene . Microarray technology has the potential for screening a large number of different antibiotic resistance genes by the relatively low-cost methods outlined in this paper.

Antimicrob Agents Chemother, 2003 Oct, 47(10), 3214 - 21
Antimicrobial resistance genes in enterotoxigenic Escherichia coli O149:K91 isolates obtained over a 23-year period from pigs; Maynard C et al.; A total of 112 Escherichia coli O149:K91 strains isolated from pigs with diarrhea in Quebec, Canada, between 1978 and 2000 were characterized for their genotypic antimicrobial resistance profiles . Tests for resistance to 10 antimicrobial agents were conducted . Resistance to tetracycline and sulfonamides was found to be the most frequent, but resistance to cefotaxime and ceftiofur was absent . An increase in the number of isolates resistant to at least three antimicrobials was observed over time . The distribution of 28 resistance genes covering six antimicrobial families (beta-lactams, aminoglycosides, phenicols, tetracycline, trimethoprim, and sulfonamides) was assessed by colony hybridization . Significant differences in the distributions of tetracycline {tet(A), tet(B), tet(C)}, trimethoprim (dhfrI, dhfrV, dhfrXIII), and sulfonamide (sulI, sulII) resistance genes were observed during the study period (1978 to 2000) . Sixty percent of the isolates possessed a class 1 integron, illustrating the importance of integrons in the epidemiology of antibiotic resistance in E . coli strains from pigs . Amplification of the integron's variable region resulted in four distinct fragments of 1, 1.3, 1.6, and 1.8 kb, with the 1.6- and 1.8-kb fragments appearing only during the last half of the study period . Examination of linkages among the different resistance genes showed a variety of positive and negative associations . Association analysis of isolates divided into two groups, those isolated between 1978 and 1989 and those isolated between 1990 and 2000, revealed the appearance of new positive resistance gene associations . Our genotypic resistance analyses of ETEC isolates from pigs indicate that many of the antibiotic resistance genes behind phenotypic resistance are not static but, rather, are in a state of flux driven by various selection forces such as the use of specific antimicrobials.

Antimicrob Agents Chemother, 2003 Oct, 47(10), 3138 - 44
Multilaboratory comparison of proficiencies in susceptibility testing of Helicobacter pylori and correlation between agar dilution and E test methods; Best LM et al.; Susceptibility testing was performed at seven Canadian microbiology laboratories and the Helicobacter Reference Laboratory, Halifax, Nova Scotia, Canada, to assess susceptibility testing proficiency and the reproducibility of the results for clarithromycin and metronidazole and to compare the Epsilometer test (E test) method to the agar dilution reference method . Control strain Helicobacter pylori ATCC 43504 (American Type Culture Collection) and 13 clinical isolates (plus duplicates of four of these strains including ATCC 43504) were tested blindly . The National Committee for Clinical Laboratory Standards (NCCLS) guidelines for agar dilution testing were followed, and the same suspension of organisms was used for agar dilution and E test . Antimicrobials and E test strips were provided to the investigators . Methods were provided on a website . Each center reported MICs within the stated range for strain ATCC 43504 . Compared to the average MICs, interlaboratory agreements within 2 log(2) dilutions were 90% (range, 69 to 100%) for clarithromycin by agar dilution, with seven very major errors {VMEs}, and 85% (range, 65 to 100%) by E test, with three VMEs . Interlaboratory agreements within 2 log(2) dilutions were 83% (range, 50 to 100%) for metronidazole by agar dilution, with six VMEs and eight major errors (MEs), and 75% (range, 50 to 94%) by E test, with four VMEs and four MEs . At lower and higher concentrations of antibiotic, E test MICs were slightly different from agar dilution MICs, but these differences did not result in errors . When a standardized protocol based on NCCLS guidelines was used, most participants in this study correctly identified clarithromycin- and metronidazole-susceptible and -resistant strains of H . pylori 93% of the time by either the agar dilution or E test method, and the numbers of errors were relatively equivalent by both methods.

Surg Neurol, 2003 Oct, 60(4), 354 - 9; discussion 359
Cerebral phaeohyphomycosis masquerading as a parafalcian mass: case report; Saberi H et al.; BACKGROUND: Cerebral phaeohyphomycosis caused by Fonsecaea pedrosoi is a rarity . However, about four cases have been reported in the literature . The disease remains mostly fatal despite employment of new treatment modalities . CASE: An 18-year-old boy presented seizures of recent onset . Two years back, he developed cutaneous phaeohyphomycosis after a splinter scratch on his chest wall . Imaging revealed a contrast enhancing parafalcian solid mass . Right frontal parasagittal craniotomy was performed and the lesion resected as much as possible, followed by IV amphotericin B and oral itraconazole treatment . The patient has been doing well during a 15-month follow-up period . DISCUSSION: Cerebral phaeohyphomycosis is an extremely rare lesion, which could masquerade as a parafalcian mass . Radical surgical removal together with antimicrobials remains the cornerstone treatment of cerebral phaeohyphomycosis.

J Gastroenterol, 2003, 38(8), 717 - 26
Antimicrobial peptides in innate immunity of the human intestine; Otte JM et al.; The intestinal mucosa has to withstand exposure to a variety of substances, challenges in pH, temperature, and osmolarity; and, finally, bacterial products which might induce local and systemic inflammatory responses . The mucosal integrity is conserved by a defense system which consisting of constitutive and inducible mechanisms . These include the physical barrier function; the secretion of factors into the lumen, such as mucins and antibacterial substances; the mucosal immune system; and, finally, the ability of the mucosa to reconstitute once damage has occurred . The homeostasis and integrity of the gastrointestinal mucosa ultimately depends upon the balance between defensive and aggressive factors . While the physical barrier function was formerly believed to play the major role in mucosal protection against luminal bacteria, the recent discovery of Toll-like receptors and antimicrobial peptides in the intestinal epithelium has modified the concept of intestinal defense towards a more active character, which will be discussed in this review.

Arch Gynecol Obstet, 2003 Oct, 268(4), 284 - 8 Epub 2002 Oct 26.
Fecundity and morbidity following acute pelvic inflammatory disease treated with doxycycline and metronidazole; Heinonen PK et al.; We studied fecundity and late sequelae of 39 women who had laparoscopic and microbiological sampling-proven acute pelvic inflammatory disease (PID) treated with the same antimicrobial regimen . The grade and etiology of index PID were classified using laparoscopy, endometrial biopsy and microbiological cultures from the cervix, endometrium and tubes: 20 had mild and 19 severe PID . The mean (SD) follow-up period after the index PID was 125 (44) {range 8-204} months . The primary end-point was pregnancy . All other or recurrent infections or other diseases related to the infection, including infertility, were evaluated . Twenty (51%) women had laparotomy or second laparoscopy during follow-up and findings were evaluated . Chlamydia trachomatis was isolated in 38% of all cases . Eleven (28%) of 39 women avoided conception or it was no longer possible . Twenty-eight women had tried to conceive after the index PID and 25 (89%) of them had at least one pregnancy . Twenty-five women had 56 pregnancies, 33 (59%) of which ended in delivery, 9 (16%) miscarried, 13 (23%) were induced abortion and only one (1.8%) tubal pregnancy occurred . Etiologic factors or severity of PID made no difference to the conception rate . Patients with mild or moderate salpingitis had a high conception rate . Endometriosis was found in 6 (30%) out of 20 women with second laparoscopy or laparotomy . Hysterectomy had been performed in 4 cases . Precise diagnosis of acute PID is the cornerstone for the treatment of the condition . Combination regimens, including drugs against the most common factors underlying acute PID against both aerobic and anaerobic microbes, prevent late sequelae in cases with mild or moderate salpingitis but not in cases with tubal or pelvic abscess.

Eur Arch Otorhinolaryngol, 2004 May, 261(5), 238 - 41 Epub 2003 Sep 18.
Expression of human beta-defensin 2 in human nasal mucosa; Chen PH et al.; Human beta-defensin (HBD)-2, an antimicrobial peptide, has been discovered to be produced by a number of epithelial cells . It is identified as a key element in the innate host defense mechanism . Because little is known about the expression of HBD-2 in the human sinonasal tract, we conducted this study to investigate the expression of the HBD-2 mRNA gene by the reverse transcription polymerase chain reaction (RT-PCR) and localization of HBD-2 peptide by immunohistochemistry in human nasal inferior turbinates and nasal polyps . RT-PCR showed significantly higher expression of HBD-2 mRNA in nasal polyps than in inferior turbinates . Using immunohistochemistry, HBD-2 peptide was predominantly localized in surface epithelial cells . Thus, it is feasible that HBD-2 is expressed in nasal mucosa and is upregulated in a condition of chronic inflammation.

Spinal Cord, 2003 Oct, 41(10), 574 - 6
Neurobrucellosis and a demonstration of its involvement in spinal roots via magnetic resonance imaging; Goktepe AS et al.; OBJECTIVE: To present a neurobrucellosis case with spinal root involvement by means of magnetic resonance imaging (MRI) . METHODS: A case of neurobrucellosis resembling Guillain-Barre syndrome is being reported . This case is unique in a way that spinal root involvement because of brucellosis was for the first time confirmed by MRI . SETTING: Spinal cord unit of a rehabilitation and care center in Ankara, Turkey . RESULTS: The correct diagnosis was made with cerebrospinal fluid culture . The patient showed a significant improvement with antimicrobial therapy and rehabilitation . CONCLUSION: Polyradiculopathy because of neurobrucellosis may mimic neurological syndromes . Rehabilitation should also be a part of its treatment.

Biochemistry, 2003 Sep 30, 42(38), 11366 - 72
The peptide antibiotic clavanin A interacts strongly and specifically with lipid bilayers; van Kan EJ et al.; In this study the interaction of the antimicrobial peptide clavanin A with phosphatidylcholine bilayers is investigated by DSC, NMR, and AFM techniques . It is shown that the peptide interacts strongly and specifically with the lipids, resulting in increased order-disorder phase transition temperatures, phase separation, altered acyl chain and headgroup packing, and a drastically changed surface morphology of the bilayer . These results are interpreted in terms of clavanin-specific interactions with lipids and are discussed in the light of the different mechanisms by which clavanin A can destroy the barrier function of biological membranes.

Curr Issues Intest Microbiol, 2003 Sep, 4(2), 43 - 51
Ionophores: their use as ruminant growth promotants and impact on food safety; Callaway TR et al.; Ionophores (such as monensin, lasalocid, laidlomycin, salinomycin and narasin) are antimicrobial compounds that are commonly fed to ruminant animals to improve feed efficiency . These antimicrobials specifically target the ruminal bacterial population and alter the microbial ecology of the intestinal microbial consortium, resulting in increased carbon and nitrogen retention by the animal, increasing production efficiency . Ionophores transport ions across cell membranes of susceptible bacteria, dissipating ion gradients and uncoupling energy expenditures from growth, killing these bacteria . Not all bacteria are susceptible to ionophores, and several species have been shown to develop several mechanisms of ionophore resistance . The prophylactic use of antimicrobials as growth promotants in food animals has fallen under greater scrutiny due to fears of the spread of antibiotic resistance . Because of the complexity and high degree of specificity of ionophore resistance, it appears that ionophores do not contribute to the development of antibiotic resistance to important human drugs . Therefore it appears that ionophores will continue to play a significant role in improving the efficiency of animal production in the future.

Vnitr Lek, 2003 Jun, 49(6), 457 - 64
{Calculation of the costs of drugs, medical materials, certain medical procedures and social services for patients with diabetic foot syndrome}; Brunerova L et al.; The aim of this study was to analyse the direct costs of some components of primary, secondary and tertiary prevention of the diabetic foot syndrome (cost of illness study) . Information considering the costs of prevention and therapy of the diabetic foot and costs following after amputation in diabetic patients is available from international economical trials but is almost missing in the Czech Republic . The economical assessment of the most important mean of primary prevention/which is regular dispensarisation and preventive care for diabetic patients in risk of diabetic foot syndrome/is very problematic . The costs of primary prevention were calculated as the intervention costs of risk factors of diabetic foot syndrome according to the recommended daily doses and prices of the medicaments in AISLP database . Costs of hospitalisation in secondary and tertiary prevention were calculated by using bills for health insurance company in the programme Steiner-UNIS, costs for out-patient care according to ordinations (medication, aids, codes for heath insurance companies and means of diagnostic process) . Costs of tertiary prevention were based on information gained from the Regional Social Authority . There were 20 patients (12 men, 8 women, age 65 +/- 12 years, 3 patients with 1 . type diabetes, 17 with 2 . type diabetes), involved in secondary in-patient care, 30 (20 men, 10 women, age 69 +/- 12.4 years, 2 patients with type 1 . diabetes, 28 with type 2 diabetes) in out-patient care and 20 (12 men, 8 women, age 70 +/- 9.4 years, 1 patient with type 1 diabetes, 19 with type 2 diabetes) in tertiary prevention . This study proved high costs of the intervention of hyperglycaemia in type 1 . diabetics, hypertension with ACE inhibitors, critical ischaemia, neuropathy . Costs of out-patient care for ulcers for eight months were 7,500.- +/- 4,400.-CK, antimicrobial treatment 1,900.-CK, in-patient care for ulcers 22,500.- +/- 6,400.-CK and amputation 22,000.- +/- 9,900.-CK . The highest costs were calculated for tertiary prevention--the first year after amputation 400,000.- CK . The study shows very high costs of tertiary prevention and suggests discussion on this topic.

Ear Nose Throat J, 2003 Aug, 82(8), 576 - 80, 82-4, 586 passim
Efficacy and tolerability of telithromycin for 5 or 10 days vs amoxicillin/clavulanic acid for 10 days in acute maxillary sinusitis; Luterman M et al.; Telithromycin (HMR 3647) is a new ketolide antimicrobial that was developed for the treatment of community-acquired respiratory tract injections . We conducted a randomized, double-blind, multicenter study to compare the clinical efficacy and safety of oral telithromycin, at 800 mg once daily for 5 or 10 days, with that of amoxicillin/clavulanic acid, at 500/125 mg three times daily for 10 days, in adults with acute maxillary sinusitis (AMS) . A total of 754 patients with AMS of less than 28 days' duration were randomized to receive either telithromycin for 5 days followed by placebo for 5 days, telithromycin for 10 days, or amoxicillin/clavulanic acid for 10 days . Clinical outcome was assessed at a test-of-cure (TOC) visit between days 17 and 24 and at a late post-therapy visit between days 31 and 45 . Analysis of clinical outcome on a per-protocol basis (n = 434) demonstrated therapeutic equivalence among the three regimens at the TOC visit; in each treatment group, the clinical cure rate was approximately 75% . Only a few patients (3 to 5 in each group) had relapsed by the late post-therapy follow-up visit . Telithromycin was generally safe and well tolerated . The most common adverse events were mild or moderate gastrointestinal effects, which occurred with similar frequency in all three groups . We conclude that 5 or 10 days of telithromycin at 800 mg once daily is as effective clinically and as well tolerated as 10 days of treatment with amoxicillin/clavulanic acid . Telithromycin, therefore, appears to be a valuable option for the treatment of AMS.

Schweiz Rundsch Med Prax, 2003 Aug 13, 92(33), 1343 - 9
{Calculated antibiotic therapy in gastroenterologic infections}; Braun RW; In gastroenterologic infections a calculated antibiotic therapy is not only determined by the suspected variety of bacterials usually associated with given symptoms but also by considerations of their potential side effects, pharmaokinetics and penetration as well as an assessment of the immune status of the patient . As some antibiotic combinations show synergistic activity whereas others are antagonistic or result in accumulated toxicity, this manuscript presents the antimicrobial profile of clinically important antibiotics including their side effects, kinetics, penetration and possible combinations . From these data recommendations for a rational and calculated antibiotic therapy of gastrointestinal infections are developed taking the guidelines of the respective scientific societies into consideration.

Arch Pharm (Weinheim), 2003 Aug, 336(8), 381 - 4
Syntheses of 2, 5-dialkylfuran and tetrahydrofuran carbinols and their cytotoxic activity; Krauss J et al.; 2, 5-Dialkylfuran and tetrahydrofuran compounds as structural elements of Annonaceae acetogenins like Asitrocinwere synthesized starting from furfural by Grignard reactions, lithiation of the furan ring and addition of aliphatic aldehydes . Hydrogenation of the furan rings over Pd-catalyst gave the corresponding tetrahydrofurans . All resulting compounds showed no or rather less antimicrobial activity against grampositive, gram-negative bacteria and fungi compared to tetracycline or clotrimazol but high cytotoxic activity against HL 60 cell line determined in the MTT assay.

Nat Rev Immunol, 2003 Oct, 3(10), 791 - 800
Macrophage migration inhibitory factor: a regulator of innate immunity; Calandra T et al.; For more than a quarter of a century, macrophage migration inhibitory factor (MIF) has been a mysterious cytokine . In recent years, MIF has assumed an important role as a pivotal regulator of innate immunity . MIF is an integral component of the host antimicrobial alarm system and stress response that promotes the pro-inflammatory functions of immune cells . A rapidly increasing amount of literature indicates that MIF is implicated in the pathogenesis of sepsis, and inflammatory and autoimmune diseases, suggesting that MIF-directed therapies might offer new treatment opportunities for human diseases in the future.

AIDS, 2003 Sep 26, 17(14), F23 - 32
alpha-Defensins can have anti-HIV activity but are not CD8 cell anti-HIV factors; Mackewicz CE et al.; BACKGROUND: CD8 T cells from healthy HIV-infected individuals inhibit HIV replication in infected CD4 T cells by a non-cytotoxic mechanism mediated by a soluble CD8 cell antiviral factor, CAF . Recently, the antimicrobial peptides, alpha-defensins, were reported to constitute CAF . OBJECTIVE: To examine the antiviral activity of alpha-defensins and address their potential role in CD8 cell non-cytotoxic antiviral responses . DESIGN AND METHODS: A purified mixture of human neutrophil proteins (HNP) 1-3 (alpha-defensins) was used to examine the effect of alpha-defensins on HIV virions and on HIV replication in CD4 cells treated prior to or post infection . alpha-Defensin expression was analyzed at the RNA and protein level in CD8 cells as well as in various other cell types . Antibodies to the defensins were tested for their ability to inhibit CAF activity in CD8 cell culture fluids . RESULTS: The alpha-defensins exhibited anti-HIV activity on at least two levels: directly inactivating virus particles; and affecting the ability of target CD4 cells to replicate the virus . However, while we could demonstrate alpha-defensins in neutrophils and monocytes, we found no evidence for the production of these peptides by CD8 T cells . No messenger RNA encoding these proteins was detected in purified CD8 T cells, nor did these cells produce intracellular or extracellular alpha-defensin peptides . Moreover, antibodies specific for human alpha-defensins 1, 2, and 3 did not block the antiviral activity of CAF-active CD8 cell culture fluids . CONCLUSIONS: The alpha-defensins are not produced by CD8 cells but unexpectedly were found to be expressed in monocytes . alpha-Defensins can have anti-HIV activity but are not CD8 cell antiviral factors.

Infect Immun, 2003 Oct, 71(10), 5488 - 97
Interleukin-18 facilitates the early antimicrobial host response to Escherichia coli peritonitis; Weijer S et al.; To determine the role of endogenous interleukin-18 (IL-18) during peritonitis, IL-18 gene-deficient (IL-18 KO) mice and wild-type mice were intraperitoneally (i.p.) infected with Escherichia coli, the most common causative agent found in septic peritonitis . Peritonitis was associated with a bacterial dose-dependent increase in IL-18 concentrations in peritoneal fluid and plasma . After infection, IL-18 KO mice had significantly more bacteria in the peritoneal lavage fluid and were more susceptible for progression to systemic infection at 6 and 20 h postinoculation than wild-type mice . The relative inability of IL-18 KO mice to clear E . coli from the abdominal cavity was not due to an intrinsic defect in the phagocytosing capacity of their peritoneal macrophages or neutrophils . IL-18 KO mice displayed an increased neutrophil influx into the peritoneal cavity, but these migratory neutrophils were less activate, as reflected by a reduced CD11b surface expression . These data suggest that endogenous IL-18 plays an important role in the early antibacterial host response during E . coli-induced peritonitis.

Peptides, 2003 Jul, 24(7), 955 - 61
Characterization of novel antimicrobial peptides from the skins of frogs of the Rana esculenta complex; Ali MF et al.; Rana esculenta is a hybridogenetic hybrid between Rana ridibunda and Rana lessonae and so is best considered as a complex of interbreeding species rather than a discrete single species . In this study, antimicrobial peptides were isolated from a pooled extract of the skins of specimens of the R . esculenta complex collected in the wild . In addition to several peptides belonging to the brevinin and esculentin families that have been previously isolated from skin secretions of a single specimen of R . esculenta, three newly described members of the brevinin-2 family (brevinin-2Ei, brevinin-2Ej, and brevinin-2Ek) and one member of the temporin family (temporin-1Ec) were purified and characterized . In addition, three structurally related peptides with no sequence similarity with antimicrobial peptides isolated from other species of ranid frogs, that potently and selectively inhibit the growth of the Gram-positive bacterium Escherichia coli (minimal inhibitory concentration (MIC<5 microM)), were identified . These peptides show limited amino acid sequence similarity to the homologous exon gene products that encode the N-terminal flanking peptides of preprocaerulein, preproxenopsin, and preprolevitide and so have been termed caerulein precursor-related fragments (CPRF-Ea, CPRF-Eb, and CPRF-Ec) . The data suggest that there may be considerable polymorphism among specimens from different populations of the R . esculenta complex . It is proposed that the distribution and amino acid sequences of skin antimicrobial peptides may be useful markers for taxonomic classification of particular sub-populations and for an understanding of phylogenetic interrelationships.

Drugs, 2003, 63(20), 2157 - 68
Appropriate empirical antibacterial therapy for nosocomial infections: getting it right the first time; Kollef M; The increasing presence of drug-resistant bacterial infections among hospitalised patients has resulted in greater numbers of patients receiving inappropriate antimicrobial treatment . This has led to the development of a novel paradigm guiding the administration of empirical antimicrobial therapy for patients with serious infections in the hospital setting . Antibacterial de-escalation is an approach to antibacterial utilisation that attempts to balance the need to provide appropriate, initial antibacterial treatment while limiting the emergence of antibacterial resistance . The goal of de-escalation is to prescribe an initial antibacterial regimen that will cover the most likely bacterial pathogens associated with infection while minimising the emergence of antibacterial resistance . Antibacterial resistance is minimised by narrowing the antibacterial regimen once the pathogens and their susceptibility profiles are determined, and by employing the shortest course of therapy clinically acceptable.

Eur Biophys J, 2004 Apr, 33(2), 109 - 16 Epub 2003 Sep 10.
Solid-state NMR study of antimicrobial peptides from Australian frogs in phospholipid membranes; Balla MS et al.; Antimicrobial peptides, isolated from the dorsal glands of Australian tree frogs, possess a wide spectrum of biological activity and some are specific to certain pathogens . These peptides have the capability of disrupting bacterial membranes and lysing lipid bilayers . This study focused on the following amphibian peptides: (1) aurein 1.2, a 13-residue peptide; (2) citropin 1.1, with 16 residues; and (3) maculatin 1.1, with 21 residues . The antibiotic activity and structure of these peptides have been studied and compared and possible mechanisms by which the peptides lyse bacterial membrane cells have been proposed . The peptides adopt amphipathic alpha-helical structures in the presence of lipid micelles and vesicles . Specifically 15N-labelled peptides were studied using solid-state NMR to determine their structure and orientation in model lipid bilayers . The effect of these peptides on phospholipid membranes was determined by 2H and 31P solid-state NMR techniques in order to understand the mechanisms by which they exert their biological effects that lead to the disruption of the bacterial cell membrane . Aurein 1.2 and citropin 1.1 are too short to span the membrane bilayer while the longer maculatin 1.1, which may be flexible due to the central proline, would be able to span the bilayer as a transmembrane alpha-helix . All three peptides had a peripheral interaction with phosphatidylcholine bilayers and appear to be located in the aqueous region of the membrane bilayer . It is proposed that these antimicrobial peptides have a "detergent"-like mechanism of membrane lysis.

Ann Hematol, 2003 Oct, 82 Suppl 2, S118 - 26 Epub 2003 Sep 09.
Diagnosis and antimicrobial therapy of pulmonary infiltrates in febrile neutropenic patients--guidelines of the Infectious Diseases Working Party (AGIHO) of the German Society of Hematology and Oncology (DGHO); Maschmeyer G et al.; Patients with severe neutropenia lasting for more than 10 days, who develop fever and pulmonary infiltrates, are at high risk of treatment failure and infection-related death, under conventional broad-spectrum antibiotics . Early supplementation by a systemic antifungal therapy active against Aspergillus spp . has been shown to markedly improve their clinical outcome . Prognosis is significantly influenced by early identification of lung infiltrates by means of high-resolution thoracic computed tomography . Non-culture based diagnostic procedures using a highly sensitive Sandwich ELISA assay to detect circulating galactomannan, or PCR techniques to amplify circulating fungal DNA, may facilitate the diagnosis of invasive pulmonary aspergillosis . CT-directed bronchoscopy and bronchoalveolar lavage using standardized procedures are useful in order to identify causative microorganisms such as filamentous fungi or Pneumocystis carinii . The standard antifungal agent in the treatment of these patients, amphotericin B deoxycholate, has been challenged recently by newly developed antifungals such as voriconazole . It seems important to continue antifungal treatment for at least 14 days before first response assessment . Microbial isolates from blood cultures, bronchoalveolar lavage or respiratory secretions must be critically interpreted with respect to their etiological relevance for pulmonary infiltrates, to avoid inadequate antimicrobial treatment modificationPublication Types:

	Guideline
	Practice Guideline

What Is Salmonella?, What Is Botulism?, What Is Prokaryote?, What Is Amino Acid?, What Is Activated Sludge?, r, Microbes, o, Bacteria, a, Microorganisms, o, Bacteriology, i, Microorganism, c, Antibiotics, n, Bactericidal, s, Wastewater, e, Culture medium, s, Cephalosporin, s, Escherichia coli, c, Staphylococcus, o, Escherichia coli, i, Achromobacter, e, Campylobacter, r, Escherichia coli, n, Antibiotics, i, Botulin, n, Meningococcus, o, Escherichia coli, a, Bacillus, c, Streptococci, o, Escherichia coli, n, Salmonella typhimurium, c, Microorganisms, a, Bacillus

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005