J Microbiol Methods, 2005 Mar, 60(3), 395 - 401 The detection of Cryptosporidium parvum and Escherichia coli O157 in UK bivalve shellfish; Macrae M et al.; Optimised immunomagnetic separation methods to detect Cryptosporidium parvum and Escherichia coli O157 in UK shellfish are described . Whole tissue homogenates gave the best recoveries for C . parvum oocysts compared with gill or haemolymph extracts . The sensitivity of recovery from spiked samples was comparable to that achieved when processing water and varied from 12-34% in mussels, 48-69.5% in oysters and 30-65% in scallops . Maximum recovery of E . coli O157 was achieved by enriching in buffered peptone water supplemented with vancomycin at 42 degrees C . Increasing enrichment temperatures from 37 to 42 degrees C gave a significant increase in target number recovery . Implementation of these methods into monitoring programmes and end-product testing will enable shellfish producers to better assess product safety. J Comb Chem, 2005 Jan-Feb, 7(1), 123 - 9 Conformational Studies of Resin-Bound Vancomycin and the Complex of Vancomycin and Ac(2)-l-Lys-d-Ala-d-Ala; Yao NH et al.; The molecular target of vancomycin, a commonly used glycopeptide antibiotic, is the d-Ala-d-Ala dipeptide subunit on the bacterial cell wall . The molecular basis of interaction between vancomycin and d-Ala-d-Ala in solution is well-known . However, there is no structural data on vancomycin, and its interaction with d-Ala-d-Ala when the drug is tethered to a solid support . In this Article, vancomycin was directly coupled onto TentaGel or PEGA resin through its C terminus . High-resolution magic angle spinning NMR studies indicated that conformation of PEGA bead-bound vancomycin is identical to that of the free drug . Broadening and shifts of the same proton resonances were observed in solution-phase vancomycin or PEGA-bound vancomycin when complexed with Ac(2)-l-Lys-d-Ala-d-Ala . This study demonstrates that bead-bound molecules can behave the same as solution-phase molecules in terms of molecular interaction with its target molecule, thus validating the on-bead screening approach of the "one-bead-one-compound" combinatorial library method. J Biol Chem . 2005 Jan 4; {Epub ahead of print} The role of the novel Fem protein VanK in vancomycin resistance in streptomyces coelicolor; Hong HJ et al.; The non-pathogenic, non-glycopeptide-producing actinomycete Streptomyces coelicolor carries a cluster of seven genes (vanSRJKHAX) that confers inducible, high-level resistance to vancomycin . The vanK gene has no counterpart in previously characterized vancomycin resistance clusters, yet vanK is required for vancomycin resistance in S . coelicolor . VanK belongs to the Fem family of enzymes, which add the branch amino acid(s) to the stem pentapeptide of peptidoglycan precursors . Upon exposure to vancomycin, the VanRS two-component system switches on expression of all seven van genes and the VanHAX enzymes reprogram the cell wall, such that precursors terminate D-Ala-D-Lac rather than D-Ala-D-Ala, thus conferring resistance to vancomycin, which only binds D-Ala-D-Ala-containing precursors . Here we provide biochemical and genetic evidence that VanK is required for vancomycin resistance because the constitutively expressed FemX enzyme, encoded elsewhere on the chromosome, cannot recognize D-Lac-containing precursors as a substrate, while VanK can . Consistent with this view, D-Lac-containing precursors carrying the Gly branch are present in the wild type transiently exposed to vancomycin, but are undetectable in a vanK mutant treated in the same way . Further, femX null mutants are viable in the presence of vancomycin but die in its absence . Because only VanK can recognise D-Lac-containing precursors, vancomycin-induced expression of VanHAX in a vanK mutant is lethal, and so vanK is required for vancomycin resistance. J Chromatogr A, 2004 Dec 10, 1060(1-2), 205 - 14 Temperature and enantioseparation by macrocyclic glycopeptide chiral stationary phases; Berthod A et al.; Seventy-one chiral compounds were separated on four macrocyclic glycopeptide chiral selectors: teicoplanin, its aglycone, ristocetin A and vancomycin, using three possible separation modes: reversed phase with methanol/buffer mobile phases, normal phase with hexane/ethanol mobile phases and polar ionic mode (PIM) with 100% methanol mobile phase with trace amounts of acid and/or base . These 148 separations were studied in a 5-45 degrees C temperature range . Peak efficiencies always increased with temperature, but in only 17% of the separations studied a small increase of the enantioresolution factor was observed . In the majority (83%) of the cases, the enantioresolution decreased or even vanished when temperature increased . All 148 Van't Hoff plots were linear showing that the selector did not change in the temperature range studied . The calculated enthalpy and entropy variations showed that the interaction of the solute with the stationary phase was always enthalpy driven with normal and reversed mobile phases . It could be enthalpy as well as entropy driven with PIM mobile phases strongly dependent on the solute . The plots of delta(deltaH) versus delta(deltaS) were linear in most cases (enthalpy entropy compensation) . This observation cannot be used to give clear information on chiral recognition mechanisms, but it allowed identifying specific stationary phase-solute interactions because the points corresponding to the respective thermodynamic parameters were clearly delineated from the general compensation lines. Anal Biochem, 2005 Jan 15, 336(2), 172 - 7 Using surface plasmon resonance to directly identify molecules in a tripeptide library that bind tightly to a vancomycin chip; Tseng MC et al.; This paper describes a procedure, based on direct binding, for identifying tight-binding ligands for a receptor immobilized on a sensor chip from an array of equimolar tripeptides using surface plasmon resonance . Vancomycin and a library of 96 tripeptides, with molecular weight ranging from 316 to 560Da, were used as a model system to illustrate the procedure . A consensus structure of the strongest interacting peptides consisted of d-Ala at the C terminus and aromatic amino acid in the penultimate position . Ligands having this structure bound more tightly to vancomycin than the known d-Ala-d-Ala peptide . The throughput of our continuous assay is 96 compounds in 3.3h, and the sample consumption is less than 2mug per peptide and 1ng for vancomycin . This procedure should be applicable to peptide libraries of greater complexity than that used here and to mixtures of small organic compounds. Rev Neurol, 2004 Dec 1-15, 39(11), 1034 - 7 {Cerebrovascular disease as an initial finding in childhood tuberculosis meningoencephalitis.}; Rotta NT et al.; INTRODUCTION . The prevalence of tuberculosis in developing countries, such as Brazil, remains high with important morbidity and mortality rates among children . Neurological complications are frequent and tuberculous meningo-encephalitis (TBM) is the most dreaded of them in infancy . CASE REPORT . Our case involves a 7-year-old white female patient who was previously in good health . Over a period of two weeks she suffered from high temperatures and vomiting and was given amoxicillin . She later suffered an attack of focal seizures . Electroencephalogram studies showed a temporary double focus and lesions in the left hemisphere . A cranial computerised tomography (CT) scan revealed a periventricular haemorrhage on the left side . The control CT (carried out 20 days later) showed a reduction in the haemorrhage and localised hypodensity . Owing to the suspected existence of an abscess, the patient was administered vancomycin . A new cranial CT scan (40 days after the first) showed gliosis alongside the basal nuclei with impregnation in the carotid trifurcation, which led us to suspect that we were dealing with a case of vasculitis or a granuloma . All the microbacterial cultures were negative and there was no history of contact with tuberculosis . The adenosine deaminase (ADA) value in the sample of cerebrospinal fluid (taken seven weeks after the first) was found to be 21.2 UI . Treatment was started with tuberculostatic compounds . Two weeks later the fever disappeared . The control CT scan showed decreased hypodensity and impregnation . The patient was discharged from hospital, with a slight monoparesis in the upper right limb . Discussion . The presentation of a cerebrovascular disease within a context of TBM, like the case reported here, is relatively rare in the literature . We concluded that the uncommon initial symptoms of TBM, associated with the negative cultures, contributed to the delay in reaching a diagnosis . A cerebrovascular accident must be included in the clinical picture of TBM and this disease has to be taken into account when dealing with a case of cerebrovascular accident. Pharmazie, 2004 Nov, 59(11), 828 - 32 HPLC enantioseparation of potential beta-blockers of the aryloxyaminopropanol type . Study of the mechanism of enantioseparation, Part VIII; Hrobonova K et al.; This paper presents the results of HPLC enantioseparation of derivatives of aryloxyaminopropanols obtained using six chiral stationary phases {macrocyclic antibiotic (vancomycin, teicoplanin, teicoplanin aglycone, and methylated teicoplanin aglycone) and cyclodextrin (beta and gamma)} and a mixture of methanol/acetonitrile/acetic acid/triethylamine (45/55/0.3/0.2) as the mobile phase . No significant difference was observed in the separation of the enantiomers on the vancomycin and teicoplanin chiral stationary phases . Comparing the separation of enantiomers on teicoplanin-based columns the retention factors were increased in the order: native teicoplanin < teicoplanin aglycone < methylated teicoplanin aglycone . The highest values of resolution were obtained on the column containing carbohydrate moieties . The presence of saccharide moieties in the chiral stationary phase plays an important role, together with charge interactions and steric interactions, in the separation of enantiomers of derivatives of aryloxyaminopropanol. J Sep Sci, 2004 Nov, 27(15-16), 1303 - 8 Enantiomeric separation of chlorophenoxy acid herbicides by nano liquid chromatography-UV detection on a vancomycin-based chiral stationary phase; Rosales-Conrado N et al.; Enantiomeric separation of mecoprop, dichlorprop, and fenoprop herbicides in their acid form, commonly used to control the growth of broad-leaved weeds, was carried out by nano-liquid chromatography (nano-LC) at a flow rate of 60 nL/min, using a packed capillary column with vancomycin-modified silica particles of 5 microm . The length of chiral stationary phase was 21 cm, while the total and effective lengths were 43 and 33cm, respectively . Inner diameter was 0.075 mm . Separated peaks were detected at 195 nm . Several mixtures of methanol, water, and 500 mM ammonium acetate buffer at different pH's were tested as mobile phase, and experimental parameters such as resolution (Rs), capacity factor (k), efficiency (N/m), and enantioselectivity factor (alpha) were measured under all the test conditions . Baseline enantiomeric separation was obtained for the three studied herbicides with alpha in the range 1.6-1.9, using as the mobile phase aqueous solutions containing 85% methanol, 5% of 500mM ammonium acetate pH4.5 buffer, and 10% water . Experimental results show that the vancomycin stationary phase presents a great enantiorecognition capability towards chlorophenoxy acid herbicides on using nano-LC. Biochemistry, 2004 Dec 14, 43(49), 15446 - 52 Synchronized conformational fluctuations and binding site desolvation during molecular recognition; Jusuf S et al.; Binding site desolvation is a poorly understood prerequisite to ligand binding . Although structural fluctuations may be expected to have an important role, little is known about which fluctuations are important or the mechanism by which they promote desolvation . This investigation examines whether and how specific structural fluctuations contribute to desolvation of the ligand binding site in glycopeptide antibiotics . Backbone peptide group rotations in vancomycin, known to occur by experimental observation, were examined in this work with a two-dimensional adaptive umbrella sampling molecular dynamics simulation technique . Results indicate that energetic barriers to rotation are relatively small for two of the peptide groups intimately involved in ligand recognition . When they occur, these rotations strip water molecules away from key hydrogen bond donors and simultaneously cause significant distortions in the macrocyclic rings of the antibiotic that force water into and out of the binding site . Both events are intricately synchronized on the molecular level and have consequences that are clearly necessary to prepare the binding site for receiving a ligand . These results suggest that previously reported observations concerning structural dynamics and binding kinetics in these compounds are mechanistically linked, and they illustrate a heretofore unrecognized degree of preorganization, complexity, and synchronization that may be involved in specific molecular recognition . They also suggest that strategies for increasing antibiotic affinity through covalent dimerization may be counterproductive. Am J Perinatol, 2004 Nov, 21(8), 433 - 8 Large vancomycin overdose in two premature infants with minimal toxicity; Miner LJ et al.; Two premature infants received 10-fold overdoses of vancomycin with resulting peak plasma concentrations > 300 microg/mL . Discontinuation of vancomycin and watchful waiting were employed, with no specific intervention to accelerate vancomycin clearance . Plasma vancomycin concentration < 40 microg/mL was attained at < 48 hours in one infant and < 72 hours in the other . Although one infant sustained a transient increase in serum creatinine to 1.4 mg/dL, no further evidence of renal, auditory, or other toxicity was detected in either infant acutely or long term . Contemporary preparations of vancomycin appear much less toxic than earlier formulations . Aggressive and invasive interventions to hasten clearance may be unnecessary following overdose in infants with normal or near-normal basal renal function, although careful surveillance for clearance and toxic effects still appear warranted. Eur J Pharm Biopharm, 2005 Jan, 59(1), 139 - 46 Effects of gamma-irradiation on trehalose-hydroxyethylcellulose microspheres loaded with vancomycin; Bartolotta A et al.; Ionizing radiation can be used as a drug sterilization technique, provided that the drug itself is not modified and that no toxic products are produced; moreover, if the irradiated product is a drug delivery system, the drug release characteristics must not be significantly altered by radiation . The aim of this work was to study the effects of sterilization by ionizing radiation on hydroxyethylcellulose/trehalose spherical micromatrices, containing the antibiotic vancomycin . Our experimental results showed that gamma-rays did not alter the chromophore groups of vancomycin (UV measurements), and did not modify the kinetic behavior of drug release from microspheres . Moreover, no significant changes in the shape and in the size distribution of microspheres were found after irradiation . The electron spin resonance (ESR) spectroscopy was proven to be a valid identification method of the executed radiation treatment, even after 5 years . The experimental results showed that the therapeutic application of the pharmacological system investigated was not compromised by irradiation, and that ESR spectroscopy can be used to distinguish irradiated from non-irradiated products. Asian J Surg, 2004 Jul, 27(3), 236 - 7 Colostomy with vancomycin administration as an effective treatment for toxic megacolon associated with fulminant pseudomembranous colitis: a case report; Haraguchi M et al.; We report a case of toxic megacolon associated with fulminant pseudomembranous colitis . A 72-year-old woman was admitted with severe dehydration and shock . Computed tomography showed evidence of diffuse thickening of the colonic wall, colonic dilatation and ascites . She underwent transverse colostomy and received postoperative vancomycin, both orally and by administration from the stoma . Her clinical situation improved dramatically following surgery . When a patient is unable to tolerate subtotal colectomy and ileostomy because of a severe overall condition, temporary colostomy followed by administration of vancomycin through the stoma is recommended. Perit Dial Int, 2004 Nov-Dec, 24(6), 590 - 5 Stability of drug additives in peritoneal dialysis solutions in a new container; Voges M et al.; OBJECTIVE: To evaluate the stability of gentamicin, tobramycin, netilmycin, vancomycin, cefazolin, unfractionated heparin, and low molecular weight heparin when added to four different peritoneal dialysis (PD) solutions {Extraneal (Baxter Healthcare, Castlebar, Ireland); Physioneal, Nutrineal, and Dianeal (Baxter Healthcare, Grosotto, Italy)} in new, non-PVC Clear-Flex containers . MEASUREMENTS: Gentamicin, tobramycin, netilmycin, vancomycin, cefazolin, unfractionated heparin, and low molecular weight heparin were injected into separate bags of PD solution . Samples were withdrawn at predefined sampling times and the concentration of each drug was analyzed using high-performance liquid chromatography (for gentamicin, tobramycin, vancomycin, and cefazolin), or bioassay (for netilmycin, gentamicin, and tobramycin in Nutrineal), or coagulation methods (heparins) . RESULTS: Netilmycin, vancomycin, cefazolin, and heparin in Physioneal, Nutrineal, Extraneal, and Dianeal were stable for at least 24 hours at 25 degrees C and for an additional 4 hours at 37 degrees C . Gentamicin in Nutrineal, Extraneal, and Dianeal was stable for at least 24 hours at 25 degrees C and for an additional 4 hours at 37 degrees C; gentamicin in Physioneal was stable for less than 24 hours at 25 degrees C . Tobramycin in Nutrineal and Extraneal was stable for at least 24 hours at 25 degrees C and for an additional 4 hours at 37 degrees C; tobramycin in Physioneal and Dianeal was stable for less than 24 hours at 25 degrees C. Biomed Chromatogr . 2004 Nov 23; {Epub ahead of print} Successful management of discovered pH dependence in vancomycin recovery studies: novel HPLC method for microdialysis and plasma samples; Plock N et al.; Vancomycin is a glycopeptide antibiotic approved for the treatment of serious infections or patients allergic to beta-lactams . A rapid HPLC assay using UV detection for the determination in microdialysate and human plasma was developed . After sample preparation, using methanol and trichloroacetic acid for plasma and water for microdialysate, 20 microL were injected and separated on a RP(18) column . Overall, the assay exhibited good precision and accuracy . The diffusion properties of vancomycin investigated in in vitro microdialysis experiments revealed an unfavourable concentration dependence avertable by keeping a constant pH using phosphate buffer as perfusate . The mean relative recoveries were 27.8% {coef fi cient of variation (CV) 11.1%} and 33.2% (CV 8.3%) for retrodialysis and recovery experiments, respectively . Following characterization of vancomycin in in vitro microdialysis, the developed setting is suitable for application in (pre-)clinical studies . Copyright (c) 2004 John Wiley & Sons, Ltd. Clin Orthop, 2004 Oct, (427), 22 - 4 Gentamicin and vancomycin do not impair experimental fracture healing; Haleem AA et al.; The purpose of this study was to determine whether gentamicin or vancomycin impair experimental fracture healing in rats . Forty-one male Wistar rats were divided into three groups, receiving either 1.5 mg/kg of gentamicin intramuscularly twice daily for 3 weeks (n = 15), 25 mg/kg of vancomycin intraperitoneally twice daily for 3 weeks (n = 14), or no treatment (n = 12), starting 7 days after production of closed, nondisplaced, bilateral femoral fractures . The mean 30 minute serum concentrations of gentamicin and vancomycin were 4.5 microg/mL and 35.1 microg/mL, respectively . Biomechanical and radiographic studies were used to evaluate fracture healing . Torsional strength testing of fracture callus exposed to gentamicin and vancomycin was similar (437 and 424 N-mm, respectively) to controls (334 N-mm) . Radiographs showed similar healing in control animals (Goldberg score, 2.3) compared with rats treated with gentamicin and vancomycin (Goldberg score, 2.6 in both groups) . Results of this study do not show impaired healing of experimental fractures after exposure to gentamicin or vancomycin (such as has been reported after exposure to quinolones). Clin Exp Dermatol, 2004 Nov, 29(6), 633 - 6 Vancomycin-induced linear IgA bullous disease presenting as toxic epidermal necrolysis; Waldman MA et al.; Linear IgA bullous dermatosis (LABD) is a rare autoimmune vesiculobullous disorder characterized by variable clinical presentations that may mimic bullous pemphigoid, dermatitis herpetiformis, cicatricial pemphigoid and erythema multiforme . A few cases of drug-induced LABD that clinically resembled toxic epidermal necrolysis (TEN) have been reported . A subset of patients with LABD have been found to be drug-induced; the most common drug being vancomycin . The diagnosis of LABD is confirmed by the presence of a linear band of IgA along the basement membrane zone on direct immunofluorescence microscopy . We report a case of a 77-year-old man who presented to us with vancomycin-induced LABD that presented clinically as TEN . He had a complete recovery over a 3-week period following discontinuation of the vancomycin and the addition of oral dapsone therapy . It is important to be aware that drug-induced LABD can mimic TEN. Clin Orthop, 2004 Oct, 1(427), 22 - 24 Gentamicin and Vancomycin Do Not Impair Experimental Fracture Healing; Haleem AA et al.; The purpose of this study was to determine whether gentamicin or vancomycin impair experimental fracture healing in rats . Forty-one male Wistar rats were divided into three groups, receiving either 1.5 mg/kg of gentamicin intramuscularly twice daily for 3 weeks (n = 15), 25 mg/kg of vancomycin intraperitoneally twice daily for 3 weeks (n = 14), or no treatment (n = 12), starting 7 days after production of closed, nondisplaced, bilateral femoral fractures . The mean 30 minute serum concentrations of gentamicin and vancomycin were 4.5 mug/mL and 35.1 mug/mL, respectively . Biomechanical and radiographic studies were used to evaluate fracture healing . Torsional strength testing of fracture callus exposed to gentamicin and vancomycin was similar (437 and 424 N-mm, respectively) to controls (334 N-mm) . Radiographs showed similar healing in control animals (Goldberg score, 2.3) compared with rats treated with gentamicin and vancomycin (Goldberg score, 2.6 in both groups) . Results of this study do not show impaired healing of experimental fractures after exposure to gentamicin or vancomycin (such as has been reported after exposure to quinolones). J Clin Microbiol, 2004 Nov, 42(11), 5429 - 31 Catheter-related bloodstream infection caused by Mycobacterium brumae; Lee SA et al.; Mycobacterium brumae is a rapidly growing environmental mycobacterial species identified in 1993; so far, no infections by this organism have been reported . Here we present a catheter-related M . brumae bloodstream infection in a 54-year-old woman with breast cancer . The patient presented with high fever (39.7 degrees C), and >1,000 colonies of M . brumae grew from a quantitative culture of blood drawn through the catheter . A paired peripheral blood culture was negative, however, suggesting circulational control of the infection . The patient was treated empirically with meropenem and vancomycin, and the fever resolved within 24 h . The catheter was removed a week later, and from the tip M . brumae was isolated a second time, suggesting catheter colonization . The organism was identified by colonial morphology, sequence analysis of the 16S rRNA gene, and biochemical tests. J Nippon Med Sch, 2004 Oct, 71(5), 340 - 4 Post-cataract surgery endophthalmitis treated with core vitrectomy: a case report; Shiwa T et al.; Postoperative endophthalmitis is one of the most serious complications after cataract surgery though its frequency may be low . We report a case with post-cataract extraction bacterial endophthalmitis treated favorably by core vitrectomy through pars plana with anterior vitrectomy cutter (A-vit) . The patient, a 72-year-old woman, presented with blurred vision 7 days after phacoemulsification and aspiration (PEA) and intraocular lens (IOL) implantation . Her initial visual acuity was counting fingers . As hypopyon and corneal edema progressed in a few hours, we decided to perform vitectomy . Firstly, we performed IOL explantation and anterior vitrectomy through the corneal stab incision with A-vit attached to the phaco machine . The inflammation, however, appeared to be severe . Secondly we performed core vitrectomy with the same cutter as we used in the first operation through pars plana as well as intravitreal injection of vancomycin on the following day . The inflammation was gradually subsided and her corrected visual acuity was recovered to 30/20 at 7 months after the vitrectomy . The results is suggest that for cataract surgeons in the facilities that are not equipped with 3-port vitrectomy machine, post-cataract extraction bacterial endophthalmitis of the emergency stage can be successfully treated by core vitrectomy through pars plana as well as intravitreal injection of antibiotics with neither vitreous shaving at the vitreous base nor artificial posterior vitreous detachment. Acta Crystallogr D Biol Crystallogr, 2004 Nov, 60(Pt 11), 1935 - 42 Epub 2004 Oct 20. Recurring main-chain anion-binding motifs in short polypeptides: nests; Milner-White EJ et al.; A novel tripeptide motif called a nest has recently been described in proteins with the function of binding anionic, or partially anionic, atoms such as carbonyl O atoms . In the present work, a search for nests in small polypeptides stored in the Cambridge Structural Database is reported . 37 unique examples were found: over half form part of hydrogen-bond arrangements resembling those in proteins, such as Schellman/paperclip loop motifs, various types of beta-turn and Asx-turns or Ser/Thr-turns, while a third are in novel situations, some involving binding to anionic groups from other molecules within the crystal complex . An example is the antibiotic vancomycin, which incorporates a prominent nest forming part of a peptide-binding site . This nest binds the carboxylate of the C-terminal D-alanine of the bacterial cell-wall precursor peptide, thereby inhibiting the final step of bacterial cell-wall synthesis . As in proteins, a number of nests occur in short peptides with an alternating glycine/L-amino-acid sequence but, uniquely to non-ribosomally synthesized short peptides, several nests within them are constructed from alternating D- and L-amino acids, and such sequences seem to specially favour nests. Rev Saude Publica, 2004 Oct, 38(5), 723 - 8 Epub 2004 Oct 18. Detection of pathogens from periodontal lesions; Malheiros Vde J et al.; OBJECTIVE: To comparatively detect A . actinomycetemcomitans and F . nucleatum from periodontal and healthy sites . METHODS: Subgingival clinical samples from 50 periodontitis adult patients and 50 healthy subjects were analyzed . Both organisms were isolated using a trypticase soy agar-bacitracin-vancomycin (TSBV) medium and detected by PCR . Conventional biochemical tests were used for bacteria identification . RESULTS: A . actinomycetemcomitans and F . nucleatum were isolated in 18% and 20% of the patients, respectively, and in 2% and 24% of healthy subjects . Among A . actinomycetemcomitans isolates, biotype II was the most prevalent . Primer pair AA was 100% sensitive in the detection of A . actinomycetemcomitans from both subject groups . Primers ASH and FU were also 100% sensitive to detect this organism in healthy subject samples . Primer pair FN5047 was more sensitive to detect F . nucleatum in patients or in healthy samples than primer 5059S . Primers ASH and 5059S were more specific in the detection of A . actinomycetemcomitans and F . nucleatum, respectively, in patients and in healthy subject samples . CONCLUSIONS: PCR is an effective tool for detecting periodontal pathogens in subgingival samples, providing a faster and safer diagnostic tool of periodontal diseases . The method's sensitivity and specificity is conditioned by the choice of the set of primers used. Drug Metab Pharmacokinet, 2004 Feb, 19(1), 68 - 75 Protective effect of inactive ingredients against nephrotoxicity of vancomycin hydrochloride in rats; Hodoshima N et al.; A generic form of vancomycin for I.V . infusion (MEEK) is more soluble and stable than the brand-name form of vancomycin hydrochloride (VCM) due to the addition of two inactive ingredients: D-mannitol and Macrogol400 (PEG400) . The aim of the present study was to compare the nephrotoxicity of MEEK with that of brand-name VCM (S-VCM) and to analyze the pharmacokinetics of these preparations . Following administration to rats at the clinical dose of 40 mg/kg, there was no difference between MEEK and S-VCM with regard to pharmacokinetics and effects on the kidneys, indicating that MEEK should be as effective as S-VCM . When administered at the nephrotoxic dose of 400 mg/kg, S-VCM caused impairment of renal function and kidney damage, and an increase of the plasma concentration due to decreased renal clearance was observed . In contrast, MEEK had virtually no effect on renal function or the kidneys and did not cause a marked change of renal clearance . These findings suggest that the inactive ingredients in MEEK play a role in reducing the nephrotoxicity of VCM. Polim Med, 2004, 34(2), 31 - 8 {Effect of corundum-vancomycin composite on soft tissue reaction}; Rusiecki M et al.; The ceramics on the basis of corundum is used for implantation in the form of porous and solid materials . The solid form was used to produce tissue endoprosthesis while porous form is mainly used to fill in the bone defects . The corundum ceramics are also known to be used as coatings for implants in orthopedics and dentistry . On the other hand there is still a need to find out the new way of treatment of the chronic bone infection, during which the traditional way of antibiotics therapy is no more effective . One of the possibly solution is to use the different biomaterials as drug carriers and in the bone surgery one of the best are porous corundum implants, being themselves of high biocompatibility, and additionally containing Vancomycin . The main target of the investigation presented in this paper was the comparative assessment of the corundum ceramics and its composite containing Vancomycin after implantation into back muscle of the total of 15 rats . During the post mortem macroscopic assessment in the tissues which surrounded the implants there were no any inflammatory neither pathological changes observed . In the microscopic findings, in early periods, the observed inflammatory tissue reaction for implants with vancomycin was significantly greater what could be explained by the high concentration of the antibiotic in the given material . On the basis of the results of macroscopic and microscopic findings we can state that the composite material of corundum ceramic containing vancomycin is of high compatibility and could be regarded as the good drug carrier. J Sep Sci, 2004 Sep, 27(13), 1109 - 14 Enantiomer separations on a vancomycin stationary phase and retention mechanism of pressurized capillary electrochromatography; Yao C et al.; Several chiral drugs, promethazine, carteolol, celiprolol, and albuterol, were resolved with vancomycin as the chiral stationary phase by pressurized capillary electrochromatography (pressurized CEC) and capillary HPLC . The effects of pressure and electrical field strength on efficiency, resolution, and capacity factor in pressurized CEC were investigated . A mathematical model describing the relationship of capacity factor in pressurized CEC with voltage, pressurized flow velocity, electroosmotic mobility, and electrophoretic mobility was established, which was in good agreement with the experimental data. Pediatr Dermatol, 2004 Sep-Oct, 21(5), 600 - 2 Extensive fixed drug eruption secondary to vancomycin; Gilmore ES et al.; Fixed drug eruption is an infrequent but well-known adverse event most commonly associated with antibiotics and nonsteroidal anti-inflammatory medications . We herein describe a second reported instance of vancomycin-induced fixed drug eruption involving an extensive area of the body surface. Ann Thorac Cardiovasc Surg, 2004 Aug, 10(4), 252 - 4 MRSA aortic valve endocarditis treated by pericardium-lined dacron patch and vancomycin-containing fibrin glue; Miyamoto S et al.; A 40-year-old man was admitted with a diagnosis of MRSA aortic valve endocarditis . He was treated conservatively with clindamycin and vancomycin for three days, but embolism occurred into the brain and the right lower limb, and urgent aortic valve replacement was performed . Resecting an aortic annular abscess resulted in a huge defect of the root . The defect was reconstructed with a combined patch: a Dacron graft lined with pericardium using vancomycin-containing fibrin glue . Although complete healing of the infected leg wound was slow, no prosthetic valve endocarditis has been detected in the 11 months since operation. Biomed Chromatogr, 2004 Nov, 18(9), 735 - 8 An automated analyzer for vancomycin in plasma samples by column-switching high-performance liquid chromatography with UV detection; Saito M et al.; An automated analyzer for vancomycin in rat plasma by column-switching high-performance liquid chromatography (HPLC) with UV detection was developed . The method includes in-line extraction of vancomycin by ion-exchange cartridge column and a separation on a reversed-phase column with UV detection at 215 nm . Plasma samples were diluted by mobile phase solution and directly injected to HPLC . Vancomycin was quantitatively recovered from rat plasma samples . The separation was completed within 15 min . The calibration curve was linear over the range from 0.5 to 100 microg/mL with the detection and quantification limits of 0.5 microg/mL (2.5 ng on column; signal-to-noise ratio = 3) . The values of precision in intra- and inter-day assays (n = 3) were less than 1.92 and 3.69%, respectively . This method does not require time-consuming pre-treatment and is suitable for the routine assay of plasma samples . 2004 John Wiley & Sons, Ltd. Chem Biol, 2004 Sep, 11(9), 1195 - 203 DpgC is a metal- and cofactor-free 3,5-dihydroxyphenylacetyl-CoA 1,2-dioxygenase in the vancomycin biosynthetic pathway; Tseng CC et al.; 3,5-Dihydroxyphenylglycine is a crucial amino acid monomer in the nonribosomal glycopeptide antibiotic vancomycin . This nonproteinogenic amino acid is constructed from malonyl-CoA by a set of four enzymes, DpgA-D, in the biosynthetic cluster . DpgC is an unusual metal-free, cofactor-free enzyme that consumes O(2) during the conversion of 3,5-dihydroxyphenylacetyl-CoA (DPA-CoA) to the penultimate intermediate 3,5-dihydroxyphenylglyoxylate (DPGx) . We show that in anaerobic incubations, DpgC catalyzes the exchange of the C(2)-methylene hydrogens of DPA-CoA at unequal rates, consistent with enzyme-mediated formation of the substrate-derived C(2)-carbanion as an early intermediate . Incubations with (18)O(2) reveal that DpgC transfers both atoms of an O(2) molecule to DPGx product . This establishes DpgC as a 1,2-dioxygenase that mediates thioester cleavage by the oxygen transfer process . These results are consistent with a DPA-CoA C(2)-peroxy intermediate, followed by enzyme-directed alpha-peroxylactone formation and collapse by O-O bond cleavage. Anal Chem, 2004 Sep 1, 76(17), 5172 - 9 Production and properties of nanoelectrospray emitters used in fourier transform ion cyclotron resonance mass spectrometry: implications for determination of association constants for noncovalent complexes; Zampronio CG et al.; Electrospray ionization (ESI) is extensively used in the analysis of biological compounds; yet some fundamental properties of this technique are not completely understood . It is widely recognized that care should be exercised when noncovalent complexes are being studied by ESI, since weak noncovalent binding can be broken or formed during the desolvation process . In the present work, spectra from the noncovalent complex, vancomycin/diacetyl-L-lysyl-D-alanyl-D-alanine, obtained from ESI and from nanoelectrospray ionization (nanoESI), have been compared . The results indicated that the milder desolvation conditions arising as a result of the smaller sizes of droplets produced in the nanoESI source attenuated effects upon weak bonds in the desolvation process . The association constant values calculated from the relative peak intensities suggest that, when using ESI, the analyzed noncovalent complex dissociated in the condensed phase during the spraying process . The influences of experimental parameters such as tip diameter and coating for nanoESI needles were investigated . Principal component analysis, a multivariate analysis method, was applied to achieve a better evaluation of the spectra obtained using different needle diameters and coatings for the analysis of the noncovalent complex vancomycin/diacetyl-L-lysyl-D-alanyl-D-alanine . It was found that 2-microm tip diameter resulted in more reproducible spectra than the larger tip diameters tested (6-20 microm) . Biomaterials, 2005 Mar, 26(9), 1043 - 52 In vivo tissue response to resorbable silica xerogels as controlled-release materials; Radin S et al.; Biodegradable, controlled-release carrier materials with non-toxic degradation products are valuable for local delivery of biologically active molecules . Previously, it was shown that room-temperature processed silica sol-gels (or xerogels) are porous, resorbable materials that can release molecules of various sizes in a controlled, time dependent manner . Previous in vitro studies also demonstrated benefits of silica xerogels as controlled-release materials for the treatment of bone infections . Herein the tissue and cell response to xerogels is documented using a subacute implantation procedure . The tissue response was correlated to composition, surface properties, resorption rate and incorporation of the antibiotic vancomycin . Ca- and P-free and Ca- and P-containing xerogels, with and without apatite (AP) surface, were used . Xerogels were implanted either as discs in a subcutaneous site, or as granules in the iliac crest of New Zealand white rabbits . The samples with surrounding tissue were retrieved after 2 and 4 weeks of implantation . Silica xerogels implanted either as discs subcutaneously or as granules in the iliac crest showed a favorable tissue response . The granules, either with or without Ca and P content, gradually resorbed over time . The resorption was accompanied by extensive trabecular bone growth and a minimal inflammatory response . Ca- and P-containing granules with an AP-surface layer showed a slower resorption rate and more extensive new bone growth than those without AP layer . Among AP-coated granules, those with incorporated vancomycin showed the most favorable tissue response . The present in vivo data together with prior in vitro data suggest that these xerogels have potential as controlled-release materials for the treatment of bone infections and as carrier materials for a variety of other applications. Electrophoresis, 2004 Aug, 25(16), 2687 - 92 Evaluation of balhimycin as a chiral selector for enantioresolution by capillary electrophoresis; Jiang Z et al.; The glycopeptide antibiotic balhimycin and its haloanalogue bromobalhimycin were evaluated as chiral selectors for enantioresolution by capillary electrophoresis . In order (i) to eliminate the adsorption of the glycopeptide antibiotics on the capillary wall, (ii) to shorten the separation time and (iii) to improve the detection sensitivity, a combined approach of the dynamic surface coating technique, the co-electroosmotic flow electrophoresis technique and the partial filling technique was employed for the enantioresolution of 16 acidic racemates . The effect of experimental parameters (plug length of the partial filling solution containing the chiral selector, selector concentration and buffer pH) on enantiorecognition was investigated . Furthermore, the enantiorecognition ability imparted by balhimycin, bromobalhimycin and vancomycin were compared . For most tested compounds, the highest enantiorecognition was obtained with balhimycin as chiral selector . Only in the case of the enantioresolution of tiaprofenic acid, vancomycin showed a superior enantiorecognition. Org Biomol Chem, 2004 Sep 21, 2(18), 2658 - 63 Epub 2004 Aug 24. Ring-closing metathesis for the synthesis of side chain knotted pentapeptides inspired by vancomycin; ten Brink HT et al.; A versatile method for the synthesis of bicyclic side chain knotted peptides inspired by vancomycin is described . The synthetic approach is based on the incorporation of a central amino acid derivative having two allylic groups-introduced by a Stille coupling-into pentapeptide 8 containing two allylated serine residues . Treatment of this bis-ring-closing metathesis precursor with 2nd generation Grubbs catalyst results in the formation of a bicyclic pentapeptide with the correct side chain to side chain connectivity pattern as observed in vancomycin: i- 2-->i, i-->i+ 2 . Modelling studies using MacroModel hint at a cavity-like structure of the bicyclic pentapeptide which may bind suitable ligands. Klin Monatsbl Augenheilkd, 2004 Aug, 221(8), 674 - 6 {Anti-infectives in eye injuries}; Behrens-Baumann W; BACKGROUND: We present the current concepts for the use of anti-infectives parallel to surgery of traumatic eye injuries . MATERIAL AND METHODS: Data from the national and international literature were collected using the PubMed, Medline, Drugs and Pharmacology, ISI, and Cochrane databanks . In addition, the experience from our tertiary center was taken into consideration . RESULTS: For palpepral and globe injuries prophylactic and therapeutic anti-infectives are recommended as supporting measures to surgical repair . These include antiseptics (PVP-iodine), antibiotics (cefuroxime, vancomycin, ceftazidime), antimycotics (voriconazole) and anti-inflammatory substances (prednisolone) . CONCLUSIONS: Several drugs are useful for the prevention and/or treatment of infection following eye injury. Br J Clin Pharmacol, 2004 Sep, 58(3), 259 - 68 Vancomycin pharmacokinetics in critically ill patients receiving continuous venovenous haemodiafiltration; DelDot ME et al.; AIMS: To investigate the pharmacokinetics of vancomycin in critically ill patients on continuous venovenous haemodiafiltration (CVVHDF), a continuous renal replacement therapy (CRRT) and to see if routine measures approximate vancomycin clearance . METHODS: Pharmacokinetic profiles (15) of initial and steady-state doses of 750 mg twice daily intravenous vancomycin were obtained from blood and ultrafiltrate samples from 10 critically ill patients in the intensive care unit, with acute renal failure on CVVHDF (1 l h(-1) dialysate plus 2 l h(-1) filtration solution; 3 l h(-1) effluent; extracorporeal blood flow 200 ml min(-1)) . RESULTS: CVVHDF clearance of vancomycin was 1.8 +/- 0.4 l h(-1) (30 +/- 6.7 ml min(-1)) . This was 1.3-7.2 times that reported previously for vancomycin using other forms of CRRT . Total vancomycin body clearance was 2.5 +/- 0.7 l h(-1) (41.7 +/- 11.7 ml min(-1)) . The clearance of vancomycin by CVVHDF was 76 +/- 16.5% of the total body clearance . CVVHDF removed approximately half the vancomycin dose during the 12-h period (A(CVVHDF) = 413 mg) . The fraction eliminated by all routes was 60% . The sieving coefficient for vancomycin was 0.7 +/- 0.1 and for urea was 0.8 +/- 0.06 . CONCLUSIONS: Vancomycin is cleared effectively by CVVHDF . Clearance was faster than other forms of CRRT, therefore doses need to be relatively high . Urea clearance slightly overestimates vancomycin clearance . The administered doses of 750 mg every 12 h were too high and accumulation occurred, as only approximately 60% of a dose was cleared over this period . The maintenance dose required to achieve a target average steady-state plasma concentration of 15 mg l(-1) can be calculated as 450 mg every 12 h. Appl Environ Microbiol, 2004 Aug, 70(8), 4899 - 905 Persistence of Mycobacterium paratuberculosis during manufacture and ripening of cheddar cheese; Donaghy JA et al.; Model Cheddar cheeses were prepared from pasteurized milk artificially contaminated with high 10(4) to 10(5) CFU/ml) and low (10(1) to 10(2) CFU/ml) inocula of three different Mycobacterium paratuberculosis strains . A reference strain, NCTC 8578, and two strains (806PSS and 796PSS) previously isolated from pasteurized milk for retail sale were investigated in this study . The manufactured Cheddar cheeses were similar in pH, salt, moisture, and fat composition to commercial Cheddar . The survival of M . paratuberculosis cells was monitored over a 27-week ripening period by plating homogenized cheese samples onto HEYM agar medium supplemented with the antibiotics vancomycin, amphotericin B, and nalidixic acid without a decontamination step . A concentration effect was observed in M . paratuberculosis numbers between the inoculated milk and the 1-day old cheeses for each strain . For all manufactured cheeses, a slow gradual decrease in M . paratuberculosis CFU in cheese was observed over the ripening period . In all cases where high levels (>3.6 log(10)) of M . paratuberculosis were present in 1-day cheeses, the organism was culturable after the 27-week ripening period . The D values calculated for strains 806PSS, 796PSS, and NCTC 8578 were 107, 96, and 90 days, respectively . At low levels of contamination, M . paratuberculosis was only culturable from 27-week-old cheese spiked with strain 806PSS . M . paratuberculosis was recovered from the whey fraction in 10 of the 12 manufactured cheeses . Up to 4% of the initial M . paratuberculosis load was recovered in the culture-positive whey fractions at either the high or low initial inoculum. Ann Allergy Asthma Immunol, 2004 Jul, 93(1), 101 - 3 A successful challenge in a patient with vancomycin-induced linear IgA dermatosis; Joshi S et al.; BACKGROUND: Linear IgA bullous dermatosis (LABD), a subepidermal, blistering skin disease, is generally believed to be idiopathic . It has been reported in association with multiple medications, including vancomycin . In each case, complete clearance of the skin lesions occurred with discontinued use of the drug . A subsequent rechallenge reproduced the eruption within hours to days . OBJECTIVE: To present a patient with vancomycin-associated LABD who underwent a successful challenge with the antibiotic 4 years after the initial reaction . METHODS: The patient developed blistering lesions over her trunk and extremities 10 days after the initiation of vancomycin for sepsis . A biopsy specimen of a skin eruption was consistent with linear IgA dermatosis . Following discontinued use of the drug, her skin lesions resolved . Four years later, she required vancomycin for osteomyelitis . RESULTS: The patient underwent a vancomycin-graded challenge of 5 doses over 5 days . On day 1, she received 10 mg, and this was increased in a semilog fashion to 1,000 mg on day 5 . She had no recurrence of her skin lesions . CONCLUSIONS: This is the first case, to our knowledge, to show a successful rechallenge in a patient with drug-associated LABD . Since the patient did not have a reaction to the challenge, it is possible that the IgA antibodies responsible for drug-induced LABD are only present transiently and diminish over time. J Clin Pharm Ther, 2004 Aug, 29(4), 351 - 7 High dose vancomycin for osteomyelitis: continuous vs . intermittent infusion; Vuagnat A et al.; OBJECTIVES: To compare the efficacy, ease of use and safety of intermittent vancomycin infusion (IVI) and continuous vancomycin infusion (CVI) in high-dose therapy of osteomyelitis . Methods: Forty-four patients with an osteomyelitis requiring vancomycin for more than 4 weeks were prospectively included, 21 receiving IVI and 23, CVI . The target serum concentration of vancomycin was 20-25 mg/L . Pharmacokinetics, adverse effects, and clinical efficacy were recorded . RESULTS: The mean daily vancomycin dosing was the same in the two groups, but the serum vancomycin concentrations (trough or plateau) were lower in the IVI group than the CVI group (21.7 +/- 9.3 and 26.0 +/- 6.1 mg/L, respectively; P < 0.0001) . The target concentrations were achieved quicker with CVI, and daily dosing was changed more frequently in the IVI group . After reaching the target, variability of vancomycin serum concentration (trough or plateau concentrations) was higher in the IVI group than in CVI group (standard deviation 7.9 mg/L vs . 5.6 mg/L, respectively; P = 0.001) . CVI did not show clinical superiority, but adverse drug effects were more frequent in the IVI group as compared with the CVI group, 9 (42.9%) and 2 (8.7%), respectively (P = 0.03) . Survival multiple regression using Cox's proportional hazard model showed that IVI (RR = 5.9, P = 0.03) and osteomyelitis of the foot (RR = 5.2, P = 0.01) were the only factors associated with adverse drug reactions leading to treatment termination . CONCLUSIONS: CVI is practical and effective, and may be a good alternative for patients requiring prolonged treatment with high vancomycin serum levels. J Am Podiatr Med Assoc, 2004 Jul-Aug, 94(4), 389 - 94 Vancomycin: an overview for the podiatric physician; Smith RG; An increased reliance on vancomycin to treat bacterial infections has led to the emergence of vancomycin-resistant organisms . The podiatric physician must select and use vancomycin with due caution . This article presents a general review of vancomycin's pharmacology, pharmacokinetics, and dosing recommendations . Literature citations of clinically based evidence regarding the development and use of vancomycin nomograms are also presented . A vancomycin dosing nomogram is introduced as an effective tool for the prescribing podiatric physician . Appropriate use of the information presented may improve patient outcomes and enable the podiatric physician to treat patients with less effort and at a lower cost. Surg Neurol, 2004 Aug, 62(2), 142 - 50; discussion 150 Therapeutic vancomycin monitoring in children with hydrocephalus during treatment of shunt infections; Bafeltowska JJ et al.; BACKGROUND: The successful treatment of shunt infections in children with hydrocephalus is still an important problem . Diagnosis of shunt colonization is often very difficult . To treat serious central nervous system (CNS) infections, intraventricular therapy with antibiotics is necessary to reach adequate cerebrospinal fluid (CSF) concentrations and eradicate the infection . For optimal management of shunt infections the concentration of administered antibiotics in CSF should be measured . The antibiotic dosing could be modified in the individual patient after pharmacokinetic studies . METHODS: In our studies, vancomycin was applied to 10 children with hydrocephalus (including 6 with a myelomeningocele) for therapeutic purposes in shunt infections . The drug was administered IV and/or intraventricularly . During treatment the concentration of vancomycin in CSF was determined by fluorescence polarization immunoassay (FPIA) method . RESULTS: Considerable differences in vancomycin concentrations after the same intraventricular antibiotic administration were observed depending on the patient . The vancomycin levels determined at study state were often much higher than the therapeutic recommended range, and the biologic half-life period (T(1/2)) of vancomycin in cerebrospinal fluid after intraventricular administration was prolonged . CONCLUSIONS: The results of our studies give information about the pharmacokinetics of vancomycin in CSF in a group of children with hydrocephalus after intraventricular administration of the drug . In our investigation, the administration of doses smaller than 5 mg/24 hours is appropriate when the removed volume of CSF will be 20 to 30 mL/24 hours. Clin Podiatr Med Surg, 2004 Jul, 21(3), 335 - 51, vi Management of osteomyelitis; Mandracchia VJ et al.; Osteomyelitis is an infection of the medullary or cortical bone that is becoming more difficult to cure with the increasing prevalence of methicillin- and vancomycin-resistant organisms . This article discusses the etiology of osteomyelitis and the effectiveness of various treatment options. Biomed Chromatogr, 2004 Jun, 18(5), 330 - 4 HPLC microdetermination of flurbiprofen enantiomers in plasma with a glycopeptide-type chiral stationary phase column; Pehourcq F et al.; A rapid and stereospecific HPLC micromethod to quantify flurbiprofen enantiomers was developed . Both flurbiprofen enantiomers and indomethacin, used as internal standard, were extracted with methylene chloride from 100 microL of acidified plasma . The resolution of the R- and S-forms was performed on a bonded vancomycin chiral stationary phase (Chirobiotic V) with 20% of tetrahydrofuran in ammonium nitrate (100 mM, pH 5) as mobile phase . Calibration curves were linear in the range 0.5-10 microg/mL for both enantiomers . A good accuracy (< or = 5%) was obtained for all quality controls, with intra-day and inter-day variation coefficients equal or less than 7.7% . Recovery of both enantiomers was found in the range 77.4-86.3% . The lower limit of quantitation was 0.25 microg/mL for both enantiomers, without interference of endogenous components . This validated micromethod has been successfully applied for quantifying R- flurbiprofen and S- flurbiprofen in rat plasma . Methods Mol Biol, 2004, 276, 153 - 68 Affinity capillary electrophoresis to examine receptor-ligand interactions; Azad M et al.; Afffinity capillary electrophoresis (ACE) is a new analytical technique that has been shown to be an efficient and accurate tool in studying biomolecular noncovalent interactions and determining binding and dissociation constants of formed complexes . ACE uses as its basis the change in migration time of a receptor upon binding to a ligand found in the electrophoresis buffer . Subsequent Scatchard analysis using noninteracting markers realizes a binding constant . Herein, ACE and three modifications in the technique, partial-filling ACE (PFACE), flow through PFACE (FTPFACE), and multiple-step ligand injection ACE (MSLIACE) are used to probe the binding of ristocetin A (Rist A) and vancomycin (Van) from Streptomyces orientalis to D-Ala-D-Ala terminus peptides and carbonic anhydrase B (CAB, E.C.4.2.1.1) to arylsulfonamides. Methods Mol Biol, 2004, 268, 199 - 205 Detection of Erysipelothrix rhusiopathiae in clinical and environmental samples; Fidalgo SG et al.; Erysipelothrix rhusiopathiae is pathogenic for both animals and humans, causing erysipelas in swine and erysipeloid in humans . In swine, disease may be either acute or chronic, resulting in the development of arthritis and endocarditis . In Japan, erysipelas remains an animal hygiene problem causing great economic loss as infected swine are disused . Human infection closely resembles that seen in swine, with both acute and chronic forms also . The most common presentation is erysipeloid, a localized cutaneous infection . In Western Australia, an erysipeloid-like infection referred to as "crayfish poisoning" occurs in lobster fishermen and handlers . A second type of presentation is a generalized cutaneous form involving lesions that progress from the initial site of infection or appear in remote areas . The third and most serious form of disease is a septicemia that is almost always linked to endocarditis . The mortality rate in Erysipelothrix endocarditis is still high (38%) and can be explained by the use of vancomycin (to which Erysipelothrix spp . are inherently resistant) as empirical therapy . Therefore, it is critical to have an early diagnosis of E . rhusiopathiae infection.Unfortunately, several problems exist with the diagnosis of E . rhusiopathiae infections by conventional cultural procedures, and these infections are often incorrectly diagnosed . First, because of their very small colony size and slow growth rates, it is difficult to isolate E . rhusiopathiae from heavily contaminated specimens . Various selective media have been described to improve the isolation of E . rhusiopathiae from contaminated specimens; however, not all contaminants are inhibited . The development of two polymerase chain reaction (PCR) methods has created an opportunity to greatly improve the efficiency with which these organisms are detected and identified . Makino et al . designed a PCR method that amplifies a 407-bp DNA fragment derived from the 16S rRNA coding sequence . The primers in this method are specific for the genus Erysipelothrix and do not differentiate between the species . A second set of primers designed by Shimoji et al . amplifies a 937-bp DNA fragment which is derived from a sequence associated with virulence of E . rhusiopathiae . These primers are specific for E . rhusiopathiae only . Shimoji et al . also utilized a selective enrichment medium based on tryptic soy broth containing ethidium bromide and sodium azide. Neurol Res, 2004 Apr, 26(3), 312 - 5 Fluctuations in vancomycin CNS tissue concentrations following intermittent and continuous infusions in the rat; Luer MS et al.; The purpose of this investigation was to compare the variability of vancomycin concentrations in the serum and CNS when administered continuously or as intermittent intravenous infusions in the rat . The hypothesis for this investigation was that the magnitude of change in serum vancomycin concentrations directly relates to the extent of vancomycin concentration fluctuations in the CNS . Microdialysis and serum sampling techniques were employed and biologic samples were analysed for vancomycin using HPLC . Over the dosing interval, the mean changes in concentrations were 71.8 +/- 9.8% and 13.6 +/- 9.3% for serum and 61.7 +/- 7.8% and 6.8 +/- 3.5% for brain extracellular fluid in the intermittent and continuously infused groups, respectively . Accordingly, the relative changes in vancomycin concentrations in brain extracellular fluid were closely associated with corresponding changes in serum concentrations (R2=0.94) . Thus, continuous intravenous administration of vancomycin results in minimal serum and CNS tissue concentration changes as compared to traditional intermittent dosing methods and allows for more consistent vancomycin concentrations in the CNS. J Pharm Biomed Anal, 2004 May 28, 35(3), 535 - 43 A validated LC method for the determination of vesamicol enantiomers in human plasma using vancomycin chiral stationary phase and solid phase extraction; Hefnawy MM et al.; An enantioseparation high performance liquid chromatographic (HPLC) method was developed and validated to determine D-(+)- and L-(-)-vesamicol in human plasma . The assay involved the use of a solid phase extraction for plasma sample clean up prior to HPLC analysis utilizing a C18 Bond-Elute column . Chromatographic resolution of the vesamicol enantiomers was performed on a vancomycin macrocyclic antibiotic chiral stationary phase (CSP) known as Chirobiotic V with a polar ionic mobile phase (PIM) consisting of methanol:glacial acetic acid:triethylamine (100:0.1:0.05 (v/v/v)) at a flow rate of 1.0 ml/min and UV detection set at 262 nm . All analyses were conducted at ambient temperature . The method was validated over the range of 1-20 microg/ml for each enantiomer concentration (R2>0.999) . Recoveries for D-(+)- and L-(-)-vesamicol enantiomers were in the ranges of 96-105% at 3-16 microg/ml level . The method proved to be precise (within-run precision ranged from 1.3 to 2.7% and between-run precision ranged from 1.5 to 3.4%) and accurate (within-run accuracies ranged from 0.8 to 3.4% and between-run accuracies ranged from 1.7 to 5.0%) . The limit of quantitation (LOQ) and limit of detection (LOD) for each enantiomer in human plasma were 1.0 and 0.5 microg/ml (S/N=3), respectively. Biomed Sci Instrum, 2004, 40, 111 - 6 Effects of sustained delivery of thymoqiunone on bone healing of male rats; Kirui PK et al.; The use of natural products as an alternative to conventional treatment in healing and treatment of various diseases has been on the rise in the last few decades . Nigella sativa, a natural herb has long been used as a natural medicine for treatment of many acute, as well as, chronic conditions . These include diabetes, hypertension and dermatological conditions . The specific objectives of this study were: (1) to successfully deliver the active component of black seed called Thymoqiunone (TMQ) at sustained level using TCPL drug delivery system; (2) to evaluate the physiological responses associated with sustained delivery of TMQ in femoral defect animal model (bone healing) . A total of 15 adult male rats were randomly divided into three equal groups . Animals in group I served as controls, animals in group II served as sham while animals in group III served as experimental group (femur defect model) . Group III animals were surgically implanted with TCPL capsule loaded with 0.02 grams of TMQ and 200 mg vancomycin . Blood samples, x-rays and body weights were collected and recorded weekly . At the end of 30 days post treatment, all animals were sacrificed and vital as well as reproductive organs were collected and analyzed histopathologically . Metabolic biochemical markers were also evaluated . The results of this study revealed the following: (i) gross anatomical observation indicated difference in healing pattern of animals in group III compared to those in group II (sham); (ii) no significant differences in all levels of cholesterol, proteins, malondialdehyde and alkaline phosphatase in all groups; (iii) no significant differences in the wet weights of vital as well as reproductive organs in all the groups . In conclusion, it appears that sustained levels of TMQ can enhance bone healing with little or no side effects on major vital and reproductive organs. Biochemistry, 2004 May 11, 43(18), 5170 - 80 Crystal structure of vancosaminyltransferase GtfD from the vancomycin biosynthetic pathway: interactions with acceptor and nucleotide ligands; Mulichak AM et al.; The TDP-vancosaminyltransferase GtfD catalyzes the attachment of L-vancosamine to a monoglucosylated heptapeptide intermediate during the final stage of vancomycin biosynthesis . Glycosyltransferases from this and similar antibiotic pathways are potential tools for the design of new compounds that are effective against vancomycin resistant bacterial strains . We have determined the X-ray crystal structure of GtfD as a complex with TDP and the natural glycopeptide substrate at 2.0 A resolution . GtfD, a member of the bidomain GT-B glycosyltransferase superfamily, binds TDP in the interdomain cleft, while the aglycone acceptor binds in a deep crevice in the N-terminal domain . However, the two domains are more interdependent in terms of substrate binding and overall structure than was evident in the structures of closely related glycosyltransferases GtfA and GtfB . Structural and kinetic analyses support the identification of Asp13 as a catalytic general base, with a possible secondary role for Thr10 . Several residues have also been identified as being involved in donor sugar binding and recognition. Anal Chem, 2004 Apr 15, 76(8), 2387 - 92 A mechanistic study of enantiomeric separation with vancomycin and balhimycin as chiral selectors by capillary electrophoresis . Dimerization and enantioselectivity; Kang J et al.; The role of the sugar moiety of glycopeptide antibiotics in chiral recognition was investigated with capillary electrophoresis . Two glycopeptide antibiotics, vancomycin and balhimycin, were employed as models since they possess the same aglycon and almost identical sugar moieties, however, with different attachment sites to the aglycon . The observed enantioselectivity of balhimycin for dansylated alpha-amino acids is 2.6 times higher than that of vancomycin . Blocking of the sugar amino group of balhimycin by N-carbamoylation reaction with KOCN led to a significantly decreased enantioselectivity compared to vancomycin, which remained almost the same upon carbamoylation . These results suggest a major role of the amino sugar together with its site of attachment to the aglycon . A dimerization-based mechanism is proposed to explain this phenomenon due to the fact that the dimerization properties of glycopeptides are similarly related to their glycosylation patterns; e.g., the dimerization constant of balhimycin is 78 times higher than that of vancomycin . Furthermore, the dimerization of glycopeptides promotes their affinity to carboxyl-containing ligands via cooperativity effects between the dimerization and the formation of glycopeptide-ligand complexes . The higher dimer stability probably leads to a more favorable conformation for chiral recognition . Thus, it is concluded that a weakened dimerization of N-carbamoylated balhimycin results in a decreased enantioselectivity. J Appl Microbiol, 2004, 96(5), 1013 - 23 RAPID detection and quantification of E . coli O157/O26/O111 in minced beef by real-time PCR; O'Hanlon KA et al.; AIM: To develop a real-time PCR detection procedure for Escherichia coli O111, O26 and O157 from minced meat . METHODS AND RESULTS: Strains (n = 8) of each of E . coli O26, E . coli O111 and E . coli O157 were inoculated at ca 10-20 CFU g(-1) into minced retail meat and enriched for 6 h at 41.5 degrees C as follows: E . coli O26 in tryptone soya broth (TSB) supplemented with cefixime (50 microg l(-1)), vancomycin (40 mg l(-1)) and potassium tellurite (2.5 mg l(-1)); E . coli O111 in TSB supplemented with cefixime (50 microg l(-1)) and vancomycin (40 mg l(-1)); E . coli O157 in E . coli broth supplemented with novobiocin (20 mg l(-1)) . DNA was extracted from the enriched cultures, and detected and quantified by real-time PCR using verotoxin (vt1 and vt2) and serogroup (O157 per gene; O26 fliC-fliA genes and O111 wzy gene) specific primers . CONCLUSIONS: The methods outlined were found to be sensitive and specific for the routine detection of E . coli O111, O26 and O157 in minced beef . SIGNIFICANCE AND IMPACT OF THE STUDY: The enrichment, isolation and detection procedures used in this study provide a rapid routine-based molecular method for the detection and differentiation of E . coli O26, O111 and O157 from minced meat. Pharmacoepidemiol Drug Saf, 1999 Oct, 8(6), 405 - 11 Evidence of inappropriate use of vancomycin in a university affiliated hospital in Brazil; de Castro MS et al.; OBJECTIVE: To evaluate the appropriateness of use of vancomycin in a University affiliated Hospital in Brazil . METHODS: One hundred sequential therapeutic courses of vancomycin were retrospectively examined through a chart review . The prescriptions were evaluated in terms of indication, use of critical process indicators, and use of outcome measurements according to an adapted version of the criteria recommended by the American Society of Health-System Pharmacists . RESULTS: The indication for use was appropriate in 39% of the cases . Critical process indicators indicated frequent inappropriate use, mainly in relation to a low frequency of bacterial cultures (54%), infrequent determination of creatinine levels prior to therapy (57%), incorrect dosage (42%), incorrect duration of therapy (63%), and infrequent determination of serum levels of vancomycin (73%) . It was impossible to evaluate outcome measurements, since data were not collected in most patients . Larger discrepancies between recommendation and practice were detected in Paediatrics and Paediatric Pneumology Services . CONCLUSION: The misuse of vancomycin in our hospital is very common according to standard guidelines, demanding new policies of control in order to diminish the possibility of the emergence of multi-resistant strains . J Chromatogr A, 2004 Apr 9, 1033(1), 91 - 9 Effect of the eluent on enantiomer separation of controlled drugs by liquid chromatography-ultraviolet absorbance detection-electrospray ionisation tandem mass spectrometry using vancomycin and native beta-cyclodextrin chiral stationary phases; Pihlainen K et al.; Enantioseparation of nine amphetamine derivatives, methorphan and propoxyphene was studied by comparing two different chiral stationary phases, macrocyclic antibiotic vancomycin and native beta-cyclodextrin (beta-CD) . Effects of 46 eluent compositions on enantioseparation in reversed-phase (RP) and polar organic phase modes were investigated . beta-CD was found to be more suitable to phenethylamines in general and vancomycin for methorphan and propoxyphene . An eluent system capable of separating the enantiomers of all phenethylamines in one run was developed . Also, systems providing competitive analysis times for enantioseparation of methorphan and propoxyphene were reported . The suitability of the eluent systems to electrospray ionisation (ESI) was discussed and methods using a tandem mass spectrometric (MS/MS) detection were developed . The suitability of chiral LC-ESI-MS/MS was tested with 14 seized drug samples . The results were in agreement with conventional non-chiral methods . Repeatability of the methods was good and limits of detection were 25-100 ng/ml for most compounds using mass spectrometric detection. J Cutan Pathol, 2004 May, 31(5), 393 - 7 Vancomycin-induced linear IgA disease manifesting as bullous erythema multiforme; Armstrong AW et al.; BACKGROUND: Vancomycin-induced linear immunoglobulin A (IgA) disease, an autoimmune, blistering disease in response to vancomycin administration, is characterized by a subepidermal, vesiculobullous eruption and linear IgA deposition along the basement membrane zone on direct immunofluorescence . CASE REPORT: We report the case of an 81-year-old man treated with vancomycin who developed diffuse erythema multiforme and tense bullae involving the palmoplantar surfaces . Discontinuation of vancomycin therapy resulted in complete resolution of this patient's cutaneous eruption . RESULTS: Biopsy of a representative skin lesion demonstrated lichenoid interface dermatitis with focal subepidermal clefting, dyskeratosis, and prominent eosinophils . Direct immunofluorescence showed linear basement membrane staining with immunoreactants to IgA; indirect immunofluorescence demonstrated the presence of circulating IgG antibodies binding in an intercellular pattern . Immunoprecipitation studies using the patient's serum revealed 210, 130, and 83 kDa target antigens . CONCLUSIONS: Presenting with an initial clinical picture suggestive of bullous erythema multiforme, this patient's subsequent clinical course and direct immunofluorescence confirm the diagnosis of linear IgA bullous disease (LABD) . His indirect immunofluorescence findings and immunoprecipitation results suggest that circulating non-IgA antibodies may represent a newly recognized immunopathologic feature of vancomycin-induced linear IgA disease, underscoring the variable and unpredictable manifestations of this drug-induced cutaneous disease. Antimicrob Agents Chemother, 2004 Apr, 48(4), 1159 - 67 Population pharmacokinetics of arbekacin, vancomycin, and panipenem in neonates; Kimura T et al.; Immature renal function in neonates requires antibiotic dosage adjustment . Population pharmacokinetic studies were performed to determine the optimal dosage regimens for three types of antibiotics: an aminoglycoside, arbekacin; a glycopeptide, vancomycin; and a carbapenem, panipenem . Eighty-three neonates received arbekacin (n = 41), vancomycin (n = 19), or panipenem (n = 23) . The postconceptional ages (PCAs) were 24.1 to 48.4 weeks, and the body weights (BWs) ranged from 458 to 5,200 g . A one-compartment open model with first-order elimination was applied and evaluated with a nonlinear mixed-effect model for population pharmacokinetic analysis . In the fitting process, the fixed effects significantly related to clearance (CL) were PCA, postnatal age, gestational age, BW, and serum creatinine level; and the fixed effect significantly related to the volume of distribution (V) was BW . The final formulas for the population pharmacokinetic parameters are as follows: CL(arbekacin) = 0.0238 x BW/serum creatinine level for PCAs of <33 weeks and CL(arbekacin) = 0.0367 x BW/serum creatinine level for PCAs of > or = 33 weeks, V(arbekacin) = 0.54 liters/kg, CL(vancomycin) = 0.0250 x BW/serum creatinine level for PCAs of <34 weeks and CL(vancomycin) = 0.0323 x BW/serum creatinine level for PCAs of > or = 34 weeks, V(vancomycin) = 0.66 liters/kg, CL(panipenem) = 0.0832 for PCAs of <33 weeks and CL(panipenem) = 0.179 x BW for PCAs of > or = 33 weeks, and V(panipenem) = 0.53 liters/kg . When the CL of each drug was evaluated by the nonlinear mixed-effect model, we found that the mean CL for subjects with PCAs of <33 to 34 weeks was significantly smaller than those with PCAs of > or = 33 to 34 weeks, and CL showed an exponential increase with PCA . Many antibiotics are excreted by glomerular filtration, and maturation of glomerular filtration is the most important factor for estimation of antibiotic clearance . Clinicians should consider PCA, serum creatinine level, BW, and chemical features when determining the initial antibiotic dosing regimen for neonates. J Antibiot (Tokyo), 2004 Jan, 57(1), 45 - 51 Ototoxicity of a new glycopeptide, norvancomycin with multiple intravenous administrations in guinea pigs; Gao W et al.; Time courses of plasma concentration of a new glycopeptide, norvancomycin (NVCM) after single intravenous (iv) and intraperitoneal (ip) injection, and the peak plasma concentrations (Cmax's) of this drug at various doses after single iv injection were determined in guinea pigs . There were significant differences in pharmacokinetic parameters between the two routes of administration and the Cmax linearly increased with the dose used increasing . Guinea pigs with normal hearing were then used to investigate ototoxic liability of NVCM after multiple intravenous administrations (54, 108, 216mg/kg, qd for 14 days) . Postrotatory vestibular nystagmus (PRN), auditory brainstem response (ABR) and electron microscopy (SEM TEM) results showed that, similarly to vancomycin, there was no functional or morphological evidence of ototoxicity of NVCM at the dose of 54, 108 mg/kg . In the high dose group (216 mg/kg), there was a 0 approximately 4 dB elevation of hearing threshold but no morphological changes . The results suggested that the ototoxicity of NVCM is absent or minimal after multiple iv administrations within this dose range. Eur J Pharm Biopharm, 2004 Mar, 57(2), 207 - 12 PLGA microspheres for the ocular delivery of a peptide drug, vancomycin using emulsification/spray-drying as the preparation method: in vitro/in vivo studies; Gavini E et al.; The aim of this study was an in vitro/in vivo investigation on poly(lactide-co-glycolide) (PLGA) microspheres as carriers for the topical ocular delivery of a peptide drug vancomycin (VA) . The microspheres were prepared by an emulsification/spray-drying technique that can be proposed as an alternative to the double emulsion method for preparation of peptide-loaded microparticles . The drug encapsulation efficiencies were close to the theoretical values (84.2-99.5%); the average particle size, expressed as dvs, was about 11 microm . The microspheres were able to modulate the in vitro drug release of VA with a behavior dependent on their composition: the highest drug content corresponded to the highest release rate . In vivo studies were carried out by assessing the pharmacokinetic profile of VA in the aqueous humor of rabbits after topical administration of aqueous suspensions of microspheres . High and prolonged VA concentrations and increased AUC values (2-fold) with respect to an aqueous solution of the drug were observed . Increasing the viscosity of the microsphere suspension by addition of a suspending-viscosizing agent (hydroxypropylcellulose) did not produce an increase of the ocular bioavailability . PLGA microspheres can be proposed as a system for ocular delivery of peptide drugs. Ophthalmic Res, 2004 Jan-Feb, 36(1), 55 - 61 Modified high-performance liquid chromatography technique for detection of vancomycin in human vitreous; Raju B et al.; PURPOSE: To standardize the technique and methodology for estimating the level of vancomycin in human vitreous using a modified high-performance liquid chromatographic method . METHODS: Sample preparation involved spiking the vitreous with known quantities of the drug followed by a brief treatment with 30% trichloroacetic acid . Samples were analyzed on a C18 reverse-phase column using a mobile phase of 50 mM phosphate buffer (pH 4.0) and 10% acetonitrile . RESULTS: Vancomycin was detected at 198 nm . It showed a retention time of 2.2 min in the vitreous . A linear increase in the area under the peak corresponding to the drug was observed with increase in concentration of vancomycin in the vitreous . The method was applied to detect the vancomycin levels in vitreous of 5 patients with exogenous endophthalmitis, 24-48 h after intravitreal injection of vancomycin 1 mg/0.1 ml . The mean concentration of vancomycin in these samples was 120.022 +/- 59.87 microg/ml (43.859-179.09 microg/ml) . The present technique allowed a quantification limit of 1 microg/ml . CONCLUSION: This technique is suitable to estimate vancomycin levels in human vitreous in a variety of clinical and experimental studies . It has the added advantages of being less expensive, simple and rapid . J Chromatogr A, 2004 Mar 5, 1028(2), 333 - 8 Improved quantification limits in chiral capillary electrochromatography by peak compression effects; Enlund AM et al.; The peak compression effect has been applied to improve quantification limits in chiral capillary electrochromatography (CEC) . A stationary phase based on the chiral selector vancomycin (Chirobiotic V) was used for separations of the enantiomers of mianserin . By adding solvents with a low dielectric constant, e.g . 2-propanol or tetrahydrofuran, to the sample solution, peak compression could be induced . The plate numbers for the minor enantiomer increased from approximately 100,000 to 1.4-1.6 million plates/m, when the composition of the mobile phase was adjusted so that the analyte eluted within either one of two system zones originating from the sample solution . A 10-fold improvement in the quantification limit for the minor enantiomer was obtained compared to elution under non-focused conditions. Int J Radiat Oncol Biol Phys, 2004 Mar 1, 58(3), 809 - 16 Renal insufficiency in patients with hematologic malignancies undergoing total body irradiation and bone marrow transplantation: a prospective assessment; Miralbell R et al.; PURPOSE: Patients with malignant hematologic disorders undergoing bone marrow transplantation (BMT) may develop renal insufficiency . A study was undertaken to assess prospectively the subclinical renal function changes with radioisotopic methods in patients undergoing BMT for hematologic malignancies . METHODS AND MATERIALS: We studied 71 patients with normal renal function undergoing BMT for various hematologic malignancies, mostly leukemias . Conditioning included chemotherapy and 12 Gy (45 patients) or 13.5 Gy (26 patients) fractionated total-body irradiation (TBI) . In 21 patients receiving 12 Gy TBI, the kidney dose was limited to 10 Gy using partial transmission blocks fabricated after renal opacification with nonionic, hypo-osmolar contrast medium . The glomerular filtration rate (GFR) and effective renal plasmatic flow (ERPF) were determined radioisotopically before conditioning and at 4, 12, and 18 months, using (51)Cr ethylene-diamine-tetra-acetic acid and (131)I ortho-iodo-hippurate, respectively . Renal insufficiency was defined as a decrease of >/=30% in GFR or ERPF compared with the baseline values . The potential influence of patient- and treatment-related variables on renal dysfunction was assessed . RESULTS: At 4 (early) and 12-18 (late) months, a >/=30% GFR drop was observed in 54% and 49% of patients and a >/=30% ERPF drop in 44% and 34% of patients, respectively . After stepwise logistic analysis, a GFR reduction at 4 months correlated significantly with age (<40 years old, worse), TBI using kidney blocks (partial kidney shielding to 10 Gy was associated with a higher rate of renal dysfunction at 4 months compared with the full TBI dose), and days of aminoglycoside/vancomycin use . An ERPF drop at 4 months was independently related with the days of amphotericin use and days of prostaglandin E(1) use (prophylaxis against hepatic venoocclusive disease) . A GFR and ERPF reduction at 12-18 months correlated with days of amphotericin use and days of prostaglandin E(1) use, respectively . CONCLUSION: Early post-BMT renal dysfunction is associated with the administration of potentially nephrotoxic drugs . An inverse correlation with the prescribed TBI dose was observed; patients whose kidneys received 10 Gy through the use of partial shielding blocks had significantly greater renal dysfunction at 4 months . The administration of potentially nephrotoxic contrast agents used in radiotherapy treatment planning may be responsible for the latter observation . Prostaglandin E(1) use correlated with a significant reduction in ERPF at both 4 and 12-18 months. Cutis, 2004 Jan, 73(1), 65 - 7 Vancomycin-induced linear IgA bullous dermatosis: morphology is a key to diagnosis; Solky BA et al.; Vancomycin-induced linear IgA bullous dermatosis (LABD) previously has been described; however, past reports have suggested that the clinical presentation is nonspecific . We present a case of vancomycin-induced LABD with a suggestive clinical presentation; specifically, groups of annularly arranged vesicles . We propose that this clinical presentation strongly suggests drug-induced LABD and should raise a clinician's suspicion of vancomycin as the offending agent . This awareness may guide the antibiotic management of the patient while the clinician awaits histopathologic correlation. Biochemistry, 2004 Feb 3, 43(4), 970 - 80 Role of the active site cysteine of DpgA, a bacterial type III polyketide synthase; Tseng CC et al.; DpgA is a bacterial type III polyketide synthase (PKS) that decarboxylates and condenses four malonyl-CoA molecules to produce 3,5-dihydroxyphenylacetyl-CoA (DPA-CoA) in the biosynthetic pathway to 3,5-dihydroxyphenylglycine, a key nonproteinogenic residue in the vancomycin family of antibiotics . DpgA has the conserved catalytic triad of Cys/His/Asn typical of type III PKS enzymes, and has been assumed to use Cys160 as the catalytic nucleophile to create a series of elongating acyl-S-enzyme intermediates prior to the C(8) to C(3) cyclization step . Incubation of purified DpgA with {(14)C}-malonyl-CoA followed by acid quench during turnover leads to accumulation of 10-15% of the DpgA molecules covalently acylated . Mutation of the active site Cys160 to Ala abrogated detectable covalent acylation, but the C160A mutant retained 50% of the V(max) for DPA-CoA formation, with a k(cat) still at 0.5 catalytic turnovers/min . For comparison, a C190A mutant retained wild-type activity, while the H296A mutant, in which the side chain of the presumed catalytic His is removed, had a 6-fold drop in k(cat) . During turnover, purified DpgA produced 1.2 equivalents of acetyl-CoA for each DPA-CoA, indicating 23% uncoupled decarboxylation competing with condensative C-C coupling . The C160A mutant showed an increased partition ratio for malonyl-CoA decarboxylation to acetyl-CoA vs condensation to DPA-CoA, reflecting more uncoupling in the mutant enzyme . The Cys-to-Ala mutant thus shows the unexpected result that, when the normal acyl-S-enzyme mechanism for this type III PKS elongation/cyclization catalyst is removed, it can still carry out the regioselective construction of the eight-carbon DPA-CoA skeleton with surprising efficiency. Curr Opin Ophthalmol, 2004 Feb, 15(1), 51 - 5 Pharmacologic considerations for cataract surgery; Tipperman R; PURPOSE OF REVIEW: A variety of options exist for perioperative, intraoperative, and postoperative medications in cataract surgery . This article reviews some of the more timely literature on "controversial" subjects in this area . RECENT FINDINGS: Recent literature may support the rationale for intracameral vancomycin at the time of cataract surgery . The recently released fluoroquinolones moxifloxacin and gatifloxacin are reviewed in terms of their clinical properties . Pharmacologic adjuncts to surgery are discussed, as is the use of intravitreal triamcinolone both at the time of cataract surgery and postoperatively . SUMMARY: Recent advances in ophthalmic pharmacology should aid the practicing anterior segment surgeon . This may affect preoperative, postoperative, and even intraoperative regimens. Ann Pharmacother, 2004 Jan, 38(1), 62 - 5 Acute delirium associated with combined diphenhydramine and linezolid use; Serio RN; OBJECTIVE: To report a case of delirium with hallucinations presumably caused by the combination of diphenhydramine and linezolid . CASE SUMMARY: A 56-year-old white man was receiving diphenhydramine 300 mg/d for 2 days to treat pruritus caused by a bullous rash possibly induced by vancomycin . He subsequently developed visual and auditory hallucinations, with erratic, aggressive behavior persisting for 3 days . Central anticholinergic syndrome was first suspected, but the long duration and exaggerated response by a patient not prone to anticholinergic toxicity suggest that a second agent may have enhanced the reaction . DISCUSSION: The pharmacodynamic properties of linezolid make this drug a likely contributor to the marked, prolonged effects experienced by this patient . The Naranjo probability scale suggests a possible relationship between the reaction and the combination of diphenhydramine and linezolid . CONCLUSIONS: Drug-induced delirium can occur with several drugs, including diphenhydramine . Linezolid has dopaminergic properties that may enhance the central nervous system effects of anticholinergics . Precautionary monitoring of mental status should be advised when concomitantly administering linezolid with drugs in this class. J Pharm Pharmacol, 2003 Dec, 55(12), 1623 - 7 Chitosan salts as nasal sustained delivery systems for peptidic drugs; Cerchiara T et al.; The aim of this study was to describe a sustained drug release system based on chitosan salts for vancomycin hydrochloride delivery . Chitosan lactate, chitosan aspartate, chitosan glutamate and chitosan hydrochloride were prepared by spray-drying technique . Vancomycin hydrochloride was used as a model peptidic drug, the nasal sustained release of which should avoid first-pass metabolism in the liver . This in-vitro study evaluated the influence of chitosan salts on the release behaviour of vancomycin hydrochloride from the physical mixtures at pH 5.5 and 7.4 . In-vitro release of vancomycin was retarded by chitosan salts and, in particular, chitosan hydrochloride provided the lowest release of vancomycin. Nephrol Dial Transplant, 2004 Feb, 19(2), 400 - 5 Evaluation of an in vitro dialysis system to predict drug removal; Hudson JQ et al.; BACKGROUND: Variation in the extent of drug removal under different dialysis conditions presents a challenge for prediction of drug elimination and dosage regimen adjustment during haemodialysis (HD) . Dependence on clinical pharmacokinetic studies in HD patients for dosing guidelines is problematic given the increasing number of dialysers with variable rates of drug removal . Thus, the purpose of this study was to characterize drug removal using an in vitro system and to evaluate its reliability to predict in vivo elimination by HD using vancomycin (VANC) as a model drug . METHODS: In vitro dialysis was performed for 2 h (volume 4.0 l normal saline, initial VANC concentration 30 mg/l, flow rate 300 ml/min, dialysate flow 800 ml/min) using four different dialysers: polymethylmethacrylate (BK-2.1 U), polysulfone (F-80), AN69 (Filtral-20) and hemophan (COBE 700HE) . The in vitro dialysis clearance for VANC (CL(D)) for the polysulfone dialyser was compared with values determined in eight HD patients . In vitro VANC CL(D) for all dialysers was compared with the clearance and KoA for B12 reported for each dialyser . RESULTS: In vitro VANC CL(D) values were 93+/-11 ml/min for the polymethylmethacrylate BK-2.1, 136+/-7 ml/min for the AN69, 65+/-9 ml/min for the hemophan COBE 700HE and 143+/-10 ml/min for the polysulfone F80 . The CL(D) for the polysulfone F80 was not statistically different from the in vivo CL(D) of 135+/-18 ml/min (P = 0.48) . In vitro VANC CL(D) correlated with the B12 CL(D) (r(2) = 0.77) and the B12 KoA (r(2) = 0.63) reported for each dialyser . CONCLUSION: VANC CL(D) in HD patients for the polysulfone dialyser was correctly predicted using the in vitro dialysis system . Use of this system may be superior to estimations of drug CL(D) based on dialyser information provided by the manufacturer for compounds of similar molecular weight. Polim Med, 2003, 33(3), 19 - 26 {Investigation of tissue reaction of apatite cement implants impregnated with vancomycin}; Juszkiewicz W et al.; Investigation of biocompatibility degree of apatite cement as a carrier of antibiotic--vancomycin was the purpose of the experimental study . Investigation of local reactions of muscular tissue was carried out on 18 rats of Wistar type, by implanting samples of apatite cement with vancomycin in the dorsal muscles . Sections of the animals and microscopic investigations were made 7, 14, 30, 90, 180 and 270 days after the implantation . Investigation of local reactions of bone tissue were carried out on 12 rabbits of New Zealand breed, by implanting the investigated samples into the femoral bone in the region of trochanter . Macroscopic, radiological and microscopic investigations were carried out 1, 3, 6 and 9 months after the surgery . The obtained results of the investigations from muscular and bone tissue, were compared to the investigations in analogical tissues of apatite cement without the medication . In macro- and microscopic investigations in the early period inflammatory reaction of muscles was noticed to be stronger around the samples with vancomycin, in comparison to the control implant . In the later period there was no inflammatory reaction and a thin fibrous connective tissue surrounded the implant and a histological picture was similar, as in cases of an implant without the antibiotic . The local reaction of bone tissue both in the control and in the investigated group (apatite cement with vancomycin) was similar . In the early period the proliferation of rich cellular connective tissue, was noticed . In the later period starting from the 3rd month, formation of young bone tissue was noticed and only locally there were focuses of fibrous tissue . The process observed after splitting of the tested materials into bone tissue, was very similar to healing processes after long bone fraction . The carried out investigations showed, that tissue reaction after implantation of apatite cement and its composite with vancomycin, was very similar and showed a high degree of tissue biocompatibility. Clin Infect Dis, 2003 Dec 15, 37(12), 1609 - 16 Epub 2003 Nov 20. Linezolid does not increase the risk of thrombocytopenia in patients with nosocomial pneumonia: comparative analysis of linezolid and vancomycin use; Nasraway SA et al.; Reports from uncontrolled studies suggest that linezolid is associated with rates of thrombocytopenia higher than those reported in clinical studies . We assessed the risk of thrombocytopenia in 686 patients with nosocomial pneumonia who received linezolid or vancomycin for > or =5 days in 2 randomized, double-blind studies and for whom follow-up platelet counts had been measured . New-onset thrombocytopenia (platelet count of <150x10(9) platelets/L) occurred in 19 (6.4%) of 295 linezolid recipients and 22 (7.7%) of 285 vancomycin recipients with baseline platelet counts of > or =150x10(9) platelets/L; severe thrombocytopenia (platelet count of <50x10(9) platelets/L) occurred in only 1 patient in each group . Platelet counts decreased to less than the baseline level in 4 (6.6%) of 61 linezolid recipients and 5 (11.1%) of 45 vancomycin recipients who had baseline counts of <150x10(9) platelets/L . No patient had a decrease to <20x10(9) platelets/L . There were no statistically significant differences between groups in these or any other platelet assessments . Clinically significant thrombocytopenia was uncommon in our analysis, and linezolid was not associated with a greater risk of thrombocytopenia in seriously ill patients than was vancomycin. J Pediatr (Rio J), 1996 Jul-Aug, 72(4), 225 - 9 {Monitorization of blood levels of vancomycin in children with multi-resistant bacterial infections}; Reis AG et al.; Pharmacokinetics of vancomycin depends on many factors and one of them is the age of the patient . It is better known in adults and there are few studies in children . Therefore, we studied the serum concentrations in 22 children with multi-resistant bacteria infections that received vancomycin . The doses administered followed the recommended rules for its utilization . This study was made at the equilibrium phase and immunofluorescence was the method utilized . The peak serum concentrations varied from 23 to 99 mcg/ml and only in 8 children (36%) they varied from 30 to 40 mcg/ml . In 10 cases (45%) we found valley serum concentration between 5 to 10 mcg/ml . These observations confirm the imprevisibility of serum concentrations obtained from one single recommended dose . Therefore, due to the complexity of vancomycin pharmacokinetics, the monitoring of blood levels is recommended in order to obtain therapeutic success. J Antimicrob Chemother, 2004 Feb, 53(2), 329 - 34 Epub 2003 Dec 19. Release of gentamicin and vancomycin from temporary human hip spacers in two-stage revision of infected arthroplasty; Bertazzoni Minelli E et al.; AIM: Evaluation of the delivery of gentamicin and vancomycin from polymethylmethacrylate (PMMA) spacers before and after implantation for the treatment of total hip replacement infections . METHODS: Twenty industrially produced spacers containing gentamicin (1.9%) were utilized . Vancomycin (2.5%) mixed with PMMA cement was used to fill holes drilled in the cement of 14 of the 20 spacers immediately before implantation . The spacers were removed from 20 patients 3-6 months after implantation and then immersed in phosphate buffer at 37 degrees C for 10 days . Antibiotic concentrations were determined by fluorescence polarization immunoassay . RESULTS: Gentamicin and vancomycin were still present in all the spacers removed from the patients . The release of gentamicin alone and in combination with vancomycin was in the range 0.05%-0.4% of the initial amount present, whereas the release of vancomycin was in the range 0.8%-3.3% . The release kinetics showed a similar pattern for both drugs . After a high initial release of drug, a reduced, but constant, elution was observed over the next few days . CONCLUSIONS: The delivery of gentamicin and vancomycin from PMMA cement was high initially, with sustained release over several months . Incorporation of vancomycin into the surface of the spacers permitted spacers to be prepared with multiple antibiotics present and without adversely affecting the release kinetics of the agents . The gentamicin-vancomycin combination shows potential for the treatment of infection following total hip replacement in specific patients. Anal Chem, 2003 Sep 15, 75(18), 4793 - 800 Determination of binding constants on microarrays with confocal fluorescence detection; Elbs M et al.; Confocal laser scanning microscopy was employed for the determination of binding constants of receptor-ligand interactions in a microarray format . Protocols for a localized immobilization of amine containing substances on glass via GOPTS (3-glycidyloxypropyl)trimethoxysilane) were optimized with respect to the detection of ligand binding by fluorescence . Compatibility with miniaturization by nanopipetting devices was ensured during all steps . The interaction of the tripeptide L-Lys-D-Ala-D-Ala with vancomycin immobilized on glass served as a model . To minimize consumption of ligand, binding constants were determined by stepwise titration of binding sites . The binding constant of the unlabeled ligand was determined by competitive titration with a fluorescently labeled analogue . The determined binding constants agreed well with those determined by other techniques, previously . Labeled ligand bound stronger than the unlabeled one . This difference was dye-dependent . Still, binding was specific for the tripeptide moiety confirming that ligand and fluorescent analogue competed for the same binding sites these results validate the determination of binding constants by competitive titration . The protocols established for confocal fluorescence detection are applicable to axially resolved detection modalities and screening for unlabeled ligands by competitive titration in general. J Appl Microbiol, 2003, 95(5), 949 - 57 Optimization of enrichment and plating procedures for the recovery of Escherichia coli O111 and O26 from minced beef; Catarame TM et al.; AIM: Optimization of enrichment media and selective agars for the detection of Escherichia coli O26 and O111 from minced beef . METHODS AND RESULTS: This study compared a number of different enrichment conditions and plating media for the recovery of E . coli O26 and E . coli O111 from minced beef . The optimum enrichment conditions for E . coli O26 was observed in beef samples enriched at 41.5 degrees C in tryptone soya broth supplemented with cefixime (50 microg l(-1)), vancomycin (40 mg l(-1)) and potassium tellurite (2.5 mg l(-1)) . Similar enrichment conditions were optimal for E . coli O111 with the omission of potassium tellurite . The optimum agar for recovery of E . coli O26 and giving the most effective suppression of contaminants was MacConkey agar {lactose replaced by rhamnose (20 g l(-1))} and supplemented with cefixime (50 microg ml(-1)) and potassium tellurite (2.5 mg l(-1)) . Optimum recovery of E . coli O111 was on chromocult agar, supplemented with cefixime (50 microg ml(-1)), cefsulodin (5 mg l(-1)) and vancomycin (8 mg l(-1)) . Minced beef samples were inoculated with a number of strains of E . coli O26 (n=9) and O111 (n=8), and the developed enrichment and plating methods, used in combination with immunomagnetic separation, were shown to be an effective method for the recovery of all strains . CONCLUSIONS: Routine cultural methods for the recovery of E . coli O26 and O111 from minced beef are described . SIGNIFICANCE AND IMPACT OF THE STUDY: The optimized enrichment and plating procedure described for the recovery of E . coli O111 and O26 from meat can be used to extend research on these emerging pathogens in beef. Nephrologie, 2003, 24(6), 287 - 92 {Drug induced linear IgA bullous dermatosis}; Montagnac R et al.; Linear IgA disease is an autoimmune subepidermal bullous disease in which linear IgA deposits are found at the basement membrane zone . It is classically idiopathic but a drug-induced variant seems to be individualized in which cutaneous lesions resolve spontaneously after cessation of responsible treatment . Among the commonly implicated drugs, vancomycin is the most frequently reported . One should not however ignore other precipitating events sometimes associated, particularly infectious diseases and non-lymphoid or lymphoproliferative malignancies . Authors present here clinical and histological features of this disease as well as drugs that have been implicated. J Food Prot, 2003 Oct, 66(10), 1911 - 5 Efficacy of enrichment broths in the recovery of freeze-injured Escherichia coli O157:H7 in inoculated ground beef by PCR; Lionberg WC et al.; Escherichia coli O157:H7 strains ATCC 35150 and ATCC 43894 and five pooled isolates from beef and pork freeze injured at -25 degrees C in beef infusion were used to inoculate ground beef . Samples (25 g each) were added to 225 ml of buffered peptone water with vancomycin, cefsulodin, and cefixime (BPW-VCC), 225 ml of modified EC broth plus novobiocin (mEC+n), and 225 ml of R&F enrichment broth (R&F-EB) and aerobically incubated at 41 to 42 degrees C . After 6, 7, 8, and 24 h of incubation, levels of E . coli O157:H7 recovered from each broth by a PCR assay with the BAX automated system as well as by conventional enrichment with the use of nonaerated mEC+n incubated at 35 degrees C for 24 h were compared with levels recovered by cultural isolation with immunomagnetic separation and plating on BCM E . coli O157:H7 chromogenic agar . For ground beef inoculated with a mean of 4.23 +/- 1.00 total cells (74% freeze injured) per 25 g, after 6 h the PCR assay identified 72.7, 57.6, and 66% of the samples for R&F-EB, BPW-VCC, and mEC+n, respectively, as presumptive positive, whereas the recovery rates after 7 and 8 h exceeded 90%, with the rate for R&F-EB being 100% . For ground beef inoculated with a mean of 1.50 +/- 0.56 total cells (80% freeze injured) per 25 g, after 6 h the PCR assay identified 47.6, 19.1, and 9.5% of the samples for R&F-EB, BPW-VCC, and mEC+n, respectively, as presumptive positive . These values increased to 81.0, 61.9, and 52.4% after 7 h and to 95.2, 61.9, and 71.4% after 8 h . After 24 h, only 55 to 60% of the samples at both inoculum levels tested positive by PCR with conventional enrichment and incubation, whereas >95% of the samples tested positive with R&F-EB aerated at 41 to 42 degrees C . Culture results for R&F-EB and mEC+n after 7 and 8 h of incubation were closely correlated with presumptive positive PCR results. Pharmacotherapy, 2003 Sep, 23(9), 1195 - 8 Vancomycin-induced thrombocytopenia: a case proven with rechallenge; Marraffa J et al.; In a rare case of vancomycin-induced thrombocytopenia, a 50-year-old man with culture-negative subacute bacterial endocarditis underwent mitral valve replacement surgery and was treated with vancomycin . His platelet count dropped from 346 x 10(3)/mm3 to 13 x 10(3)/mm3 on postoperative day 4, and a differential diagnosis of heparin- versus drug-induced thrombocytopenia was considered . Antiheparin antibodies were detected in the patient's serum on day 5 . He showed no signs of bleeding . His platelet count remained below 5 x 10(3)/mm3 despite two platelet transfusions on day 5 . A hemorrhagic pericardial effusion with tamponade developed, requiring drainage . A trial with intravenous immunoglobulin led to fever and chills, and the infusion was not completed . Vancomycin was changed to clindamycin on day 9, and methylprednisolone therapy was started on day 11 . On day 12, the patient's clinical condition improved, and his platelet count increased from 3 x 10(3)/mm3 to 32 x 10(3)/mm3 with no bleeding . On day 18, his platelet count was 424 x 10(3)/mm3, and he was scheduled for discharge with vancomycin therapy for a total of 6 weeks . He received a single dose of intravenous vancomycin 1 g at the hospital; his platelet count dropped to 160 x 10(3)/mm3 1 hour after the infusion and to 58 x 10(3)/mm3 12 hours later . Vancomycin was discontinued and clindamycin and prednisone were restarted . On day 20, the patient's platelet count increased to 105 x 10(3)/mm3 and he was discharged with warfarin, prednisone, and clindamycin therapy . We suspect that our patient's thrombocytopenia was due to vancomycin. Rev Invest Clin, 2003 May-Jun, 55(3), 276 - 80 {Pharmacologic clinical monitoring of serum vancomycin levels in pediatric patients}; Nandi-Lozano E et al.; OBJECTIVE: To analyze the results from the clinical monitoring of serum vancomycin levels among pediatric patients receiving this drug . METHODS: Retrospective study of data from routine monitoring of serum vancomycin concentrations . The study population includes children who received vancomycin for more than 3 days and had peak and trough vancomycin serum concentration documented . The vancomycin concentrations were measured by an immunoassay procedure . RESULTS: We obtained 70 vancomycin concentrations from pediatric patients whose ages ranged from newborn to 13 years old . Mean peak and trough concentrations were 45.7 +/- 8.0 and 18.7 +/- 9.6 micrograms/mL, respectively . We used as a reference normal ranges for Cpmax of 25-40 micrograms/mL and Cpmin of 5-40 micrograms/mL . Only 26.1% of the patients were on Cpmax normal ranges, 47.8% were above the range and 26% under the lower limit . In regards to the Cpmin 17% of the cases were between the accepted limits, 41% above the upper limit and 41% under the normal range . CONCLUSION: Our data suggest that less than a fifth of the patients have serum levels on therapeutic range and almost half of these population had serum levels above the normal range . This might be explained by the type of the population sampled, reflecting a selection bias by detecting levels only among patients with an increased risk for toxicity . Finally, we stress the importance of accurately documenting dose, timing, and renal function in the records of all patients subjected to serum vancomycin determinations. J Infus Nurs, 2003 Sep-Oct, 26(5), 278 - 84 Vancomycin: new perspectives on an old drug; Hadaway L et al.; The use of vancomycin continues to be prevalent in all clinical settings . However, many questions persist about infusion techniques . According to the Infusion Nursing Standards of Practice, peripheral catheters are not the best choice for infusing this drug because of its pH . The key to reducing risk of peripheral phlebitis and extravasation injury is choosing a more appropriate vascular access device . Many healthcare providers correlate systemic side effects with the infusion rate and concentration, although many reports cannot support this correlation . New technologies of vascular access and infusion controlling devices are changing old, established practices . This update provides an examination of the current literature on all aspects of infusing vancomycin and monitoring patients. J Chromatogr B Analyt Technol Biomed Life Sci, 2003 Sep 25, 795(1), 115 - 21 Role of the vancomycin-ristocetin heterodimerization on the enantioselectivity of D,L-tryptophan and D,L-dansyl tryptophan; Slama I et al.; In this paper, a chromatographic system using immobilized ristocetin as chiral stationary phase and vancomycin as chiral mobile phase additive (CMPA) was described in order to investigate the role of t