Farmaco, 2000 Sep-Oct, 55(9-10), 619 - 23 Synthesis and antimicrobial activity of new adamantane derivatives II; Orzeszko A et al.; A series of new derivatives of adamantane was synthesised . The new compound 4-(adamant-1-ylethylenoxycarbonyl)phthalanhydride obtained from 1-adamantaneethanol and trimellitic anhydride chloride, as well as 4-(adamant-1-ylmethylenoxycarbonyl)phthalanhydride, appeared very useful for preparation of a number of N-substituted phthalimides . Antimicrobial activity of newly obtained derivatives such as, for example, 4-(adamant-1-ylethylenoxycarbonyl)-N-(L-phenylalanyl)phthalimide or 4-(adamant-1-ylmethylenoxycarbonyl)-N-(L-leucyl)-phthalimide was tested against Staphylococcus aureus, Bacillus sp., Micrococcus flavus and Enterococcus faecium . The minimal inhibitory concentrations for these compounds against Bacillus cereus were 15 and 8 microg/ml, respectively. FEMS Microbiol Lett, 2001 Jan 1, 194(1), 77 - 82 Involvement of multiple genetic loci in Staphylococcus aureus teicoplanin resistance; Bischoff M et al.; Teicoplanin resistance was transformed from a teicoplanin-resistant Staphylococcus aureus into the susceptible strain BB255 to give strain BB938 . The cell wall composition, amidation of the iD-glutamate, and peptide crosslinking were identical in BB938 as in BB255 except for a 60% increased length of the glycan chain . Transductional crosses revealed that at least two distinct loci contributed in a cumulative fashion to teicoplanin resistance . One of these loci correlated with a mutation inactivating the anti-sigma factor RsbW . This mutation must have occurred during transformation and selection for teicoplanin resistance in BB938 . Genetic manipulations involving the sigB operon showed that transcription factor SigB contributed to decreased teicoplanin susceptibility. J Cardiovasc Surg (Torino), 2000 Oct, 41(5), 715 - 9 Treatment of recurrent postoperative mediastinitis with granulated sugar; De Feo M et al.; BACKGROUND: The authors report their experience with granulated sugar as dressing technique in the treatment of postoperative mediastinitis refractory to a closed irrigation system . METHODS: Between January 1990 and January 1998, mediastinitis developed in 61 (0,93%) of 6521 patients who had undergone open heart surgery . Diagnosis of sternal infections was based on wound tenderness, drainage, cellulitis, fever associated with sternal instability . All of them were initially treated with surgical debridement and closed chest irrigation . Nine patients with postcardiotomy mediastinitis refractory to closed chest irrigation underwent open dressing with granulated sugar . All of them were febrile with leukocytosis and positive wound cultures . RESULTS: Bacteria isolated were staphylococcus aureus in 6 cases, staphylococcus epidermidis in 2 and pseudomonas in 1 . Redebridement was performed in all cases and the wound was filled with granulated sugar four times a day . Fever ceased within 4.3+/-1.3 days from the beginning of treatment and WBC became normal after 6.6+/-1.6 days . Three patients had hyperbaric therapy as associated treatment . Complete wound healing was achieved in 58.8+/-32.9 days (three patients underwent successful pectoralis muscle flaps) . CONCLUSIONS: Sugar treatment is a reasonable and effective option in patients with mediastinitis refractory to closed irrigation treatment . It may be used either as primary treatment or as a bridge to pectoralis muscle flaps. Radiol Med (Torino), 2000 Sep, 100(3), 112 - 9 {Radiologic diagnosis of spondylodiscitis: role of magnetic resonance}; Cusmano F et al.; PURPOSE: To report the Magnetic Resonance Imaging (MRI) features of acute and chronic spontaneous spondylodiscitis as well as any typical patterns which can be useful for the differential diagnosis between pyogenic and tuberculous forms . MATERIAL AND METHODS: Eleven patients affected with spontaneous spondylodiscitis were selected for the study; they were 7 men and 4 women ranging in age 33-87 years (mean: 64) . We excluded the patients with iatrogenic spondylodiscitis . MR images were acquired with a superconductive magnet at 1.5, with the following sequences: sagittal PD and T2-weighted TSE, sagittal T1-weighted SE, axial PD and T2-weighted TSE for the lumbar spine, axial T2-weighted GRE for the cervical and dorsal spine and axial and sagittal T1-weighted SE after contrast agent (gadolinium DTPA) injection . MR images were reviewed by three experienced radiologists and morphological and signal intensity changes of vertebral body and disk were recorded on a standard form . In 9 patients it was possible to compare MR to CT findings . RESULTS: At the time of our observation all patients reported pain at the spine level, associated with fever and weight loss in 50% of cases and with increased values of the inflammatory markers . Three patients had infectious diseases in other organs and 2 were diabetics . Biopsy was performed in two cases only and demonstrated Staphylococcus aureus in one and Mycobacterium tuberculosis in the other patient . MRI allowed the correct diagnosis to be made in all cases, demonstrating the pathological involvement of the paravertebral structures and into the spinal canal earlier and more accurately than CT . A common finding in pyogenic and tuberculous spondylodiscitis was the low signal of the subcortical bone marrow on T1-weighted sagittal images, which enhanced after Gd-DTPA administration and became intermediate or high on T2-weighted images . Moreover, the steady high signal intensity of the disk on T2-weighted images and its contrast enhancement on T1-weighted images is typical for an acute inflammatory process . CONCLUSIONS: Based on our personal experience and literature data, we believe MRI to be the most sensitive technique for the diagnosis of spondylodiscitis in the acute phase, whereas it is comparable to CT in the chronic stage of the disease . At present MRI does not allow to differentiate pyogenic from tuberculous forms. Curr Opin Rheumatol, 2001 Jan, 13(1), 3 - 11 Management of the ear, nose, and throat manifestations of Wegener granulomatosis: an otorhinolaryngologist's perspective; Rasmussen N; A diagnosis of Wegener granulomatosis requires granulomatous manifestations in the respiratory tract . With the increasing use of antineutrophil cytoplasmic autoantibodies as a diagnostic tool, Wegener granulomatosis is diagnosed earlier than in the past, and not infrequently when only ear, nose and throat manifestations are present, placing the otorhinolaryngologist in a central role in diagnosis and management . Diagnostic biopsies should be obtained from active lesions in the nose and paranasal sinuses and concomitant infection should be identified . Because of the apparent relation between infection and activation of disease, the management of infections-especially those due to Staphylococcus aureus-requires special attention . The increasing numbers of early cases identified warrants further investigations of whether less toxic treatment regimens will be of advantage in such cases . Medical and surgical treatment of the acute and chronic manifestations presents specific problems because of altered immune competence, prevalent superinfection, and tissue destruction, and is therefore best taken care of by specially dedicated otorhinolaryngologists. Med Dosw Mikrobiol, 2000, 52(3), 229 - 36 {Methicillin resistant clinical strains of Staphylococcus aureus isolated from patients in the State Clinical Hospital Nr 2 in Szczecin}; Bilska I et al.; Over a fivefold increase, from 11% to 58%, in the prevalence of methicillin-resistance was observed in 1994-95 amongst clinical isolates of Staphylococcus aureus in the State Clinical Hospital No 2 in Szczecin, one of the largest hospitals in the West Pomeranian region of Poland . The aim of this study was to see if any one particular strain was responsible for this apparent outbreak . Fifty-six randomly selected isolates were typed by SmaI macrorestriction analysis using PFGE and by analysis of antimicrobial susceptibility patterns . Results indicate the presence of two epidemic multi-drug resistant MRSA strains . Over 85% of typed MRSA belonged to the first strain, which was probably present in the hospital long before 1994 . MRSA of this strain were isolated from patients in 8 hospital wards . The second strain was introduced into two wards of the hospital in the last year of the study. Med Dosw Mikrobiol, 2000, 52(3), 217 - 22 {Use of PCR for evaluating detection of coa and nuc genes in methicillin-resistant, coagulase-negative strains of Staphylococcus aureus (MRSA-CN)}; Mlynarczyk A et al.; Strains showing a negative reaction in tube test for coagulase constitute 10 to 20% of all Staphylococcus aureus isolated from patients of Hospital of Infant Jesus in Warsaw . Most of them are MRSA . In 42 MRSA strains showing negative reaction for coagulase, the presence of coagulase (coa) and nuclease (nucA) genes was checked . Determination of whole cell DNA with PCR reaction was performed . The obtained results revealed that all 42 strains possessed gene nucA, but only 39 strains possessed the coa gene. Ann Transplant, 2000, 5(3), 13 - 9 Anastomotic infections in lung transplant recipients; Hadjiliadis D et al.; OBJECTIVES: Anastomotic infections are an uncommon but potentially devastating complication after lung transplantation . The incidence, microbiology, predisposing factors, and clinical outcomes of anastomotic infections have not been well described . METHODS: We performed a retrospective chart review of the first 283 lung or heart-lung transplant recipients performed at Duke University Medical Center and identified all cases of anastomotic infection . RESULTS: Fifteen patients (5.3%) developed anastomotic infections . Aspergillus caused infection in six patients, Candida in eight patients and Staphylococcus aureus in one patient . Bilateral or right lung transplantation and the use of induction immunosuppression with monoclonoal or polyclonal antibodies are associated with a higher incidence of anastomotic infections . All patients with fungal anastomotic infections were treated with a combination of systemic and inhaled antifungal agents . All patients had improvement of their anastomotic sites after treatment and no patients developed anastomotic dehiscence . CONCLUSIONS: Anastomotic infection is an infrequent complication after lung transplantation, and is caused predominately by fungal pathogens . In contrast to previous reports, anastomotic dehiscence did not occur in any patient in our series . Treatment with the combination of inhaled and systemic antimicrobial agents may have favorably affected clinical outcomes. Am J Nephrol, 2000 Nov-Dec, 20(6), 463 - 7 Use of pulsed-field gel electrophoresis in the analysis of recurrent Staphylococcus aureus infections in patients on continuous ambulatory peritoneal dialysis; Chang HR et al.; BACKGROUND/AIM: The purpose of this study was to evaluate pulsed-field gel electrophoresis (PFGE) for distinguishing between relapse and reinfection of Staphylococcus aureus infections in patients on continuous ambulatory peritoneal dialysis (CAPD) . METHODS: Between July 1993 and May 1997, 4 patients with recurrent CAPD-associated infections caused by S . aureus we enrolled in this study . There were nine episodes of peritonitis, one episode of temporary double lumen catheter infection, and one episode of Hickman catheter infection . A total of eleven S . aureus isolates were collected from peritoneal fluid (n = 9) and blood (n = 2) . PFGE typing was applied . RESULTS: In our study, from PFGE typing, the 11 S . aureus isolates were classified into seven patterns . Antibiogram profiling classified only four patterns . Patient A had a reinfection by another strain of S . aureus, and patient B had three episodes of peritonitis caused by the same strain of S . aureus due to exit site infections . Patient C had two episodes of CAPD peritonitis caused by two different strains, respectively . Patient D had four episodes of S . aureus infection (three CAPD peritonitis and one bacteremia); the first two episodes of peritonitis were caused by an identical strain of S . aureus, whereas the subsequent two infections were caused by other organisms . CONCLUSION: PFGE has a high discriminatory power and can be an assistant method to antibiogram profiling for distinguishing relapse from reinfection in CAPD-associated peritonitis . Am J Emerg Med, 2001 Jan, 19(1), 1 - 5 Bacterial counts in experimental, contaminated crush wounds irrigated with various concentrations of cefazolin and penicillin; Lammers R et al.; The objective of this study was to determine if three different concentrations of cefazolin and penicillin irrigation solutions reduce quantitative bacterial counts in experimental crush wounds contaminated with multiple species of bacteria . The design used was a randomized, blinded, experimental animal study . An animal bite wound model was created by innoculating crushed incisions with three species of bacteria . Four paravertebral incisions extending to deep fascia were created in each of twelve anesthetized albino guinea pigs . Wound edges were clamped with a hemostat for five seconds to create crushed, devitalized tissue within each wound . Wounds were inoculated with 0.4 mL of a standard solution of Staphylococcus aureus, Bacterioides fragilis, and Pasturella multocida and covered . Four hours after inoculation, each wound was scrubbed for 30 seconds with 20% poloxamer 188 and then irrigated with 100 mL of one of four solutions: normal saline solution (control); cefazolin (CZ) 2 mg/mL, plus penicillin G (PCN) 200 units/mL (low dose); CZ 10 mg/mL, plus PCN 2,000 units/mL (intermediate dose); and CZ 50 mg/mL, plus PCN 20, 000 units/mL (high dose) . Investigators were blinded to the solutions used . Wounds were covered with a vapor-permeable dressing . Six days after treatment, each wound was examined for signs of infection and then excised for quantitative bacteriologic analysis . Colony counts were reported as counts per gram of tissue . Wounds in the four irrigation solution groups were compared using ANOVA . A log difference of 3 was considered significant . The average log total bacteria/gram tissue for the four groups were: control, 4.35 (95% CI; 1.01); low dose, 4.09 (95% CI; 1.42); intermediate dose, 4.47 (95% CI; 1.27); and high dose, 3.45 (95% CI; 1.33) . No wounds in the high-dose group had any clinical signs of infection, whereas 50% of wounds in the intermediate dose group, 42% in the low dose group, and 33% in the control group had either erythema, induration, or purulence . There were no statistically significant differences in the bacterial counts/gram tissue or clinical infection rates in any of the groups . A formal trend analysis failed to find a significant linear trend for decreasing bacterial counts for either antibiotic . In this experimental bite wound model containing contaminated, crushed tissue, irrigation with various solutions of cefazolin plus penicillin G did not reduce quantitative bacterial counts more than 3.1 log total bacteria/gram tissue. Microbiol Immunol, 2000, 44(11), 945 - 7 Comparative study of Staphylococcus aureus isolated from lesional and non-lesional skin of atopic dermatitis patients; Matsui K et al.; The skin of patients with atopic dermatitis (AD) is often colonized by Staphylococcus aureus, and superantigenic exotoxins produced by the organism are thought to be an important precipitating factor of AD . However, there are few reports comparing the characteristics of S . aureus isolated from the lesional and non-lesional skin of identical AD patients . In this study, therefore, we examined whether the presence of superantigen-producing S . aureus correlates with the formation of eczematous lesion of AD patients . The detection rate of S . aureus on the lesional skin of AD patients was higher than on the non-lesional skin of AD patients . Furthermore, the bacterial cell count of S . aureus on the lesional skin of AD patients was also significantly higher than that of the non-lesional skin of AD patients . However, there was no significant difference between the detection rate of superantigenic exotoxin-producing S . aureus on the lesional and nonlesional skin of AD patients . These results suggest that the number of S . aureus present is more important in the formation of eczematous lesion of AD patients than the presence of superantigenic exotoxin-producing S . aureus strains per se. Microb Drug Resist, 2000 Fall, 6(3), 253 - 8 Similarity of antibiotic resistance patterns and molecular typing properties of methicillin-resistant Staphylococcus aureus isolates widely spread in hospitals in New York City and in a hospital in Tokyo, Japan; Aires de Sousa M et al.; One hundred and forty-three single-patient methicillin-resistant Staphylococcus aureus (MRSA) isolates collected during April-June, 1997, and February, 1998, in a hospital in Tokyo, Japan, were characterized by molecular typing techniques that involved hybridization of ClaI restriction digests with the mecA- and Tn554-specific DNA probes and determination of macrorestriction patterns of SmaI-digested chromosomal DNA by pulsed-field gel electrophoresis (PFGE) . A large proportion (76%) of the isolates carried the mecA polymorph I, Tn554 pattern A, and PFGE pattern A (clonal type I:A:A), which was the same as the clonal type of an MRSA widely spread in hospitals in New York City and hospitals in neighboring New Jersey, Connecticut, and Pennsylvania . Also similarly to the New York clone, most of the MRSA isolates from the Japanese hospital were resistant to penicillin, ciprofloxacin, erythromycin, tetracycline, and high concentrations (500 microg/ml) of spectinomycin, but were susceptible to chloramphenicol, sulfamethoxazole-trimethoprim, and rifampin . All of the 143 MRSA isolates had vancomycin MICs < or = 2 mg/L. Microb Drug Resist, 2000 Fall, 6(3), 245 - 51 Distribution of methicillin-resistant Staphylococcus aureus clones among health care facilities in Connecticut, New Jersey, and Pennsylvania . ; Roberts RB et al.; A previous surveillance study conducted in 12 hospitals in New York City in 1996 identified a unique multidrug-resistant genetic lineage of methicillin-resistant Staphylococcus aureus (MRSA) that was widespread and accounted for as much as 42% of all the MRSA isolates . The purpose of the study described here was to determine possible geographic spread of this New York clone of MRSA to neighboring states . Single-patient MRSA isolates (258) from 29 health care facilities in Connecticut (CT), New Jersey (NJ), and Pennsylvania (PA) were collected during the calendar year 1998 . DNA typing, consisting of fingerprinting of chromosomal macrorestriction patterns generated by SmaI digestion followed by pulsed-field gel electrophoresis (PFGE), identified 22 patterns . PFGE type A, closely related to the PFGE type of the previously identified New York clone, accounted for 154 (60%) of 258 isolates . The clone was detected in all facilities, was predominant in 19 of the 29 health care centers, and accounted for 92% of the MRSA isolates collected in PA . The overwhelming majority of MRSA with PFGE type A was also resistant to erythromycin, ciprofloxacin, and clindamycin . One of the two most common PFGE subtypes detected in the three states sampled (PFGE subtype A1) had an identical PFGE pattern to that of the previously described vancomycin-resistant strain of S . aureus (VISA) recently detected in a hospital in Westchester, NY . The second most frequent MRSA clone with PFGE type E and accounting for 26% (68/258 isolates), also described earlier in the 12 New York City hospitals, was resistant not only to erythromycin, ciprofloxacin, and clindamycin, but also to gentamicin and sulfamethoxazole-trimethoprim as well . The unique multidrug resistance pattern of this second clone and its geographic distribution accounted for the differences observed in the frequency of multidrug resistance among MRSA isolates recovered in the three states . The pandemic Iberian clone recently detected in New York City was not detected among the 258 MRSA isolates recovered in CT, NJ, and PA. Microb Drug Resist, 2000 Fall, 6(3), 239 - 44 Resistance rather than virulence selects for the clonal spread of methicillin-resistant Staphylococcus aureus: implications for MRSA transmission; Shopsin B et al.; The population structure of methicillin-resistant Staphylococcus aureus (MRSA) is predominantly clonal, which may be related to the fitness of the genetic background of the methicillin-susceptible S . aureus (MSSA) into which the mecA chromosomal resistant determinant has inserted . To test this idea, we assessed whether the genotypes of New York MRSA are present in MSSA populations by using a combination of protein A gene sequence typing (spa typing) and pulsed-field gel electrophoresis (PFGE) . Although about 16% of colonizing MSSA isolated from community subjects were related to MRSA, only one of the five predominant New York MRSA clonal types was found among the MSSA isolates . Similarly, among nosocomial MSSA, only four MRSA homologues were observed, two of which may have arisen through deletion of the mec element . Thus, MRSA clonal types represent a limited spectrum of the diversity seen in community and hospital S . aureus populations . The data are best explained by antibiotic selection pressure, as opposed to increased transmissibility or virulence, being responsible for the clonal dissemination of the resistance phenotype in MRSA genetic backgrounds, an in turn, the limited spread of these strains outside of the hospital environment. Microb Drug Resist, 2000 Fall, 6(3), 231 - 8 Methicillin-resistant Staphylococcus aureus: phylogenetic relatedness between European epidemic clones and Swiss sporadic strains; Blanc DS et al.; We have compared the phylogenetic diversity of methicillin-resistant Staphylococcus aureus (MRSA) strains from Switzerland and their phylogenetic relationships with European epidemic clones, using multiprimer random amplification polymorphic DNA (RAPD) . Strains included 24 European epidemic clones (59 strains), 66 sporadic strains isolated in Switzerland in 1996-1997, and 15 reference strains of five other Staphylococcus species . Similarity and clustering analysis with the Jaccard's coefficient showed that the maximum genetic distance between MRSA strains was 0.43, whereas the minimum genetic distance between the six Staphylococcus species was 0.97, indicating that the method permits phylogenetic hierarchization . The 24 MRSA clones reported to be epidemic in European countries during the 1990s were distributed into seven different genetic clusters with a maximum distance of 0.29 among them . This clustering pattern was confirmed by the analysis of a subset of MRSA strains by multilocus enzyme electrophoresis at 12 loci . Most of the sporadic Swiss strains were distributed into these seven different genetic clusters, together with the epidemic MRSA clones . This suggests that there is no phylogenetic cluster specific to epidemic clones of MRSA. Microb Drug Resist, 2000 Fall, 6(3), 223 - 9 Methicillin-resistant Staphylococcal aureus evolution in Australia over 35 years; Turnidge JD et al.; Australia has a long association methicillin-resistant Staphylococcus aureus (MRSA) . Its unique geographic and demographic features have led to the emergence and spread of three types of MRSA over 35 years . Classical multiresistant hospital-acquired MRSA were first noted in Australia in 1965 . By the end of the 1970s, strains of this type of MRSA were well established in the complex tertiary care hospitals in the capital cities on the eastern seaboard of mainland Australia . Characterized by resistance to beta-lactams, erythromycin, tetracycline, gentamicin, and trimethoprim-sulfamethoxazole, these strains have persisted and diversified genetically and have acquired a variety of new resistances . They have proven pathogenicity and are a prominent cause of hospital infection in the endemic institutions . More recently they have become endemic in some central state tertiary care hospitals . Community-acquired strains of MRSA first appeared in the north of Western Australia in the mid-1980s . Strains have subsequently appeared in the south of the state and in the two adjacent central states, and are more frequently isolated from Aboriginal patients . Although harboring few or no additional resistances apart from resistance to beta-lactams initially, these strains are also accumulating additional resistances . A different variety of community-acquired MRSA has recently been noted in eastern Australia . It has a similar antibiogram to the western strains, but an entirely different epidemiology, resembling that currently being experienced in parts of New Zealand, and associated with patients of south Pacific island origin. Microb Drug Resist, 2000 Fall, 6(3), 213 - 21 Genetic relatedness of multidrug-resistant, methicillin (oxacillin)-resistant Staphylococcus aureus bloodstream isolates from SENTRY Antimicrobial Resistance Surveillance Centers worldwide, 1998; Diekema DJ et al.; We reviewed Staphylococcus aureus bloodstream infection isolates from SENTRY centers worldwide during 1998 to evaluate the molecular epidemiology of multiply drug-resistant methicillin (oxacillin)-resistant S . aureus (MDR-MRSA) . MDR-MRSA was defined as a S . aureus isolate with a MIC for oxacillin at >2 microg/ml and with four or more additional resistances . A total of 325 unique patient isolates of MDR-MRSA from five continents were analyzed using ribotyping and pulsed-field gel electrophoresis (PFGE) . The frequency of MDR-MRSA among all S . aureus BSI isolates ranged from only 2.2% in Canada to 35.6% in the Asia-Pacific region . Forty-eight ribotypes (RT) were distinguished, but over 80% of the isolates were contained within the 10 most prevalent RTs . The most common RT, RT 184.5, which included 30% of all MDR-MRSA, was found on four of five continents . PFGE provided superior discrimination and identified numerous clusters of possible clonal dissemination of MDR-MRSA within individual medical centers and between institutions that are in geographic proximity . In four instances, strains with indistinguishable PFGE patterns were found on more than one continent . The predominant PFGE subtype in South America (RT 893.5/Ia) was isolated from patients at centers in Brazil, Argentina, and Portugal, and closely related subtypes were isolated in Chile and Italy . There is great geographic variation in rates of methicillin- and multidrug-resistance among S . aureus bloodstream isolates worldwide . Although many MDR-MRSA strains group geographically, a few closely related epidemic strains have wide regional and even global range. Microb Drug Resist, 2000 Fall, 6(3), 189 - 98 Molecular typing of methicillin-resistant Staphylococcus aureus by pulsed-field gel electrophoresis: comparison of results obtained in a multilaboratory effort using identical protocols and MRSA strains; Chung M et al.; Pulsed-field gel electrophoresis (PFGE) has become the gold standard of molecular methods in epidemiological investigations . In spite of its high resolving power, use of the method has been hampered by inadequate laboratory-to-laboratory reproducibility . In the project described here we have addressed this problem by organizing a multilaboratory effort in which the same bacterial strains (subtype variants of the Iberian and Brazilian methicillin-resistant Staphylococcus aureus--MRSA--clones) were analyzed by twenty investigators in thirteen different laboratories according to an indentical protocol, which is reproduced here in detail . PFGE patterns obtained were analyzed at a central laboratory in order to identify specific technical problems that produced substandard macrorestriction patterns . The results including the specific technical problems and their most likely causes are described in this communication . Also listed are seven major epidemic clones of MRSA which have been characterized by molecular fingerprinting techniques and the prototypes of which have been deposited at the American Type Culture Collection, from where they will be available for interested investigators for the purpose of typing MRSA isolates . It is hoped that this communication will contribute to the improvement of the reproducibility and technical/aesthetic quality of PFGE analysis. Microb Drug Resist, 2000 Fall, 6(3), 173 - 88 Molecular epidemiology of methicillin-resistant Staphylococcus aureus strains: state of affairs and tomorrow' s possibilities; van Belkum A; Methicillin-resistant strains of Staphylococcus aureus (MRSA) have posed a clinical threat for nearly 40 years . During these years, an array of additional technologies suited for identification of MRSA below the species level has become available . The technologies, whether they assess phenotype or genotype, provide data that can be used for elucidation of the routes of dissemination of individual MRSA types . This review summarizes the current state of affairs with respect to the quality of the various laboratory techniques and includes descriptions of novel strategies such as binary typing and multilocus sequence typing (MLST) . Drawbacks of procedures will be compared, and the value of molecular typing in the elucidation of complex biological phenomena, such as epidemicity, carriage, and reduced vancomycin susceptibility, will be indicated . Means for integrated assessment of bacterial biology, epidemiology, and population structure will be discussed. Pediatr Infect Dis J, 2000 Dec, 19(12), 1163 - 6 Current trends in community-acquired methicillin-resistant Staphylococcus aureus at a tertiary care pediatric facility; Hussain FM et al.; BACKGROUND: The prevalence of community-acquired methicillin-resistant Staphylococcus aureus (MRSA) infections increased at the University of Chicago Children's Hospital (UCCH) from 10 per 100,000 admissions from 1988 to 1990 to 259 per 100,000 admissions from 1993 to 1995 . Because this increase may have represented a one time occurrence or a limited disease outbreak, we updated our previous observations at UCCH in 1998 and 1999 to see whether this trend had continued . DESIGN: Prospective observational study . RESULTS: Twenty-three hospitalized children had an MRSA isolate during the 1-year study period . Ten were community-acquired, equally distributed between children with predisposing risk factors and those without . The overall prevalence of community-acquired MRSA was 208 per 100,000 admissions . Seven of the 10 community-acquired MRSA isolates were susceptible to clindamycin . Skin and soft tissue infections predominated among the children with a community-acquired MRSA isolate . Pulsed field gel electrophoresis of the 10 community-acquired MRSA isolates revealed 8 distinct patterns; these data suggest that multiple clones were circulating at UCCH . CONCLUSION: MRSA are no longer confined to children with established risk factors . The prevalence of community-acquired MRSA among children without identified risk factors is high in our institution. J Indian Med Assoc, 2000 Jul, 98(7), 368 - 70 Potential infection hazards of stethoscopes; Sood P et al.; This study was conducted to assess the bacterial flora carried on stethoscopes used by medical personnel and to study the effect of disinfection of stethoscopes on the flora . In the 106 stethoscopes sampled, Gram-positive organisms were the most (60%) frequently isolated . Among them, Staphylococcus aureus accounted for 15.8% of the flora of which 21% were resistant to methicillin . The rate of isolation of S aureus and methicillin resistant S aureus (MRSA) was higher in critical care units . Disinfection was found to significantly reduce the bacterial count. Can J Microbiol, 2000 Dec, 46(12), 1108 - 14 Assessing and analysing contamination of a dairy products processing plant by Staphylococcus aureus using antibiotic resistance and PFGE; Tondo EC et al.; A dairy product processing plant was studied for 2.5 years to examine contamination with Staphylococcus aureus and try to correlate the source of contamination . Cultures were submitted to an antibiotic susceptibility test (AST) and characterised by Pulsed-field Gel Electrophoresis (PFGE) analysis . Results showed that 35.2% (19/51) of food handlers were asymptomatic carriers of S . aureus, and that 90.4% (19/21) of raw milk sampled was contaminated . Staphylococcus aureus was isolated from only 10 samples among more than 3200 investigated dairy products . No S . aureus contamination was found on machinery . The AST analysis demonstrated sensitivity of tested S . aureus to oxacillin, cephalothin, vancomycin, gentamicin, and sulfamethoxazole/trimethoprim . AST analysis generated eight different phenotypic profiles, but did not allow us to identify the source of contamination in seven of ten final products . PFGE analysis proved to be a sensitive method as it generated 42 different DNA banding profiles among the 48 S . aureus investigated, demonstrating a lack of predominance of endemic strains in the plant, contrary to suggestions raised by antibiotic resistance typing . Based on PFGE genotyping, S . aureus strains isolated from four contaminated final products were similar to four S . aureus isolated from raw milk . Five final products contained S . aureus different from all other strains collected, and one showed similarity to a strain isolated from a food handler . These results suggest contamination by raw milk as the main source of contamination of the final dairy products. Am J Perinatol, 2000, 17(8), 423 - 7 Maternal-fetal staphylococcal infections: a series report; Andre P et al.; The objective of this paper is to study the characteristics of maternal-fetal staphyloccocal infection . A retrospective study among 1,582 infants admitted consecutively to our neonatal intensive care unit was carried out from January 1995 through September 1998 . The antenatal history, and the clinical and bacteriological findings and outcome of the infants fulfilling maternal-fetal staphyloccocal infection were analysed . Among 122 (7.7%) maternal-fetal infection, 11 cases (8.9%) of congenital staphyloccal infections were diagnosed in 9 premature and 2 full-term babies . Antenatal invasive procedures were noted in 6 occasions (56%) . All the 11 infants developed respiratory and hemodynamic failure . Staphylococcus aureus was the most common organism encountered in maternal bacteriologic data (9/11, 82%) as well as on peripheral sites (9/11, 82%) and in blood cultures (7/11, 64%) performed in the infants . There was one case of methicillin-resistant Staphylococcus aureus . The outcome was favorable for 9 infants . Two very preterm neonates died within the first 72 hours of life . Mother-to-infant transmission of Staphylococcus should be suspected whenever invasive procedures are performed during pregnancy and in the presence of severe neonatal distress associated with an inflammatory response . Prompt and adapted antibiotic therapy allows a favourable outcome. Schweiz Med Wochenschr, 2000, Suppl 125, 48S - 51S {Residual bacterial contamination of rhinoscopes used in ENT consultation after cleaning with a pad impregnated with a disinfectant}; Kutter J et al.; INTRODUCTION: The duration of the official disinfection procedure of a Hopkins rhinoscope agreed by the hospital hygiene authorities may be a handicap in a busy ENT consultation . A shortened procedure is sometimes used, but without risk assessment . OBJECTIVE: To determine the risk of contamination of the endoscope used for rhinoscopy after simple manual cleaning with a pad impregnated with a disinfectant solution (Neosabenyl) . PATIENTS AND METHOD: In 60 patients undergoing rhinoscopy at our ENT policlinic, a nasal swab as well as the endoscope were examined bacteriologically . The nasal physiological flora and specifically Staphylococcus aureus were semiquantitatively analysed on each specimen . In 50 patients, the rhinoscopes were simply cleaned with an impregnated pad, whereas in 10 patients the official disinfection procedure with glutaraldehyde was applied . RESULTS: 25 out of the 50 endoscopes (50%) which were manually cleaned with the impregnated pad, were still contaminated with bacteria . Among the 12 (20%) identified healthy carriers of S . aureus, 7 (58%) showed a rhinoscope still contaminated with S . aureus after cleaning with the pad . The overall risk of contaminating the following patient with S . aureus is 14% . None of the 10 endoscopes cleaned and disinfected with glutaraldehyde were contaminated . CONCLUSIONS: Simple manual cleaning and disinfection with an impregnated pad is insufficient and carries the risk of contaminating the following patient . This emphasises the need to follow the recommended cleaning and disinfection procedures even in a small or busy practice. Schweiz Med Wochenschr, 2000, Suppl 125, 41S - 43S {Infectious aspects of Wegener's granulomatosis}; Rusterholz D et al.; INTRODUCTION: Because of manifestation of Wegener's granulomatosis in the upper respiratory tract the ENT-specialist is often initially involved in diagnosis . Recent research results suppose the chronic nasal carriage of Staphylococcus aureus triggering relapse rate in Wegener's granulomatosis . The adequate therapy of this bacteria as chronic nasal carriage may reduce the number of relapses in patients with Wegener's granulomatosis in remission . PATIENT: An illustrative case report shows the positive effect of prolonged treatment with cotrimoxazole in a 49-year-old male with a second relapse of Wegener's granulomatosis . RESULTS: The prolonged treatment of cotrimoxazole reduced the dose of cyclophosphamid and glucocorticoids which show long-term side effects . DISCUSSION: 15 years ago a positive effect of cotrimoxazole to Wegener's granulomatosis was discussed . Later immunological and clinical studies confirmed this fact . Prolonged treatment with cotrimoxazole seems to reduce the number of relapses in patients with this chronic disease . Corresponding with our case reduction of the systemic therapy with cyclophosphamid and glucocorticoids is possible. Infect Control Hosp Epidemiol, 2000 Dec, 21(12), 761 - 4 An outbreak of pyodermas among neonates caused by ultrasound gel contaminated with methicillin-susceptible Staphylococcus aureus; Weist K et al.; OBJECTIVE: To investigate an outbreak of methicillin-susceptible Staphylococcus aureus (MSSA) infections in a neonatal clinic . DESIGN: Prospective chart review, environmental sampling, and genotyping by two independent methods: pulsed-field gel electrophoresis (PFGE) and randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) . A case-control study was performed with 31 controls from the same clinic . SETTING: A German 1,350-bed tertiary-care teaching university hospital . RESULTS: There was a significant increase in the incidence of pyodermas with MSSA; 10 neonates in good physical condition with no infection immediately after birth developed pyodermas . A shared spatula and ultrasound gel were the only identified infection sources . The gel contained MSSA and was used for hip joint sonographies in all neonates . PFGE and RAPD-PCR patterns from 6 neonates and from the gel were indistinguishable and thus genetically related clones . The case-control study revealed no significant risk factor with the exception of cesarean section (P=.006) . The attack rate by days of hip-joint sonography between April 15 and April 27, 1994, was 11.8% to 40% . CONCLUSIONS: Inappropriate hygienic measures in connection with lubricants during routine ultrasound scanning may lead to nosocomial S . aureus infections of the skin . To our knowledge this source of S . aureus infections has not previously been described. Kansenshogaku Zasshi, 2000 Nov, 74(11), 966 - 72 Susceptibility to vancomycin of methicillin-resistant Staphylococcus aureus isolated in a university hospital in Japan; Mori N et al.; Intravenous vancomycin was approved in 1991 in Japan and has been widely used for treatment of infections caused by methicillin-resistant Staphylococcus aureus (MRSA) . Consequently, ever since the initial discovery of vancomycin intermediate-resistant S . aureus in Japan, the vancomycin resistance of this organism has been a great concern in clinical settings . We investigated whether vancomycin resistance had emerged in MRSA isolated in our hospital since the approval of the use of intravenous vancomycin . Vancomycin susceptibility was evaluated on the basis of minimum inhibitory concentrations determined by the agar dilution method and a heterogeneous resistance examination . The median minimum inhibitory concentration of the 69 MRSA strains isolated in 1988 and the 74 isolated in 1998 was 0.75 microgram/ml and 1.0 microgram/ml, respectively (p < 0.001), however, all of the strains were classified in the susceptible group . None of them was an MRSA heterogeneously resistant to vancomycin (hetero-VRSA), which has been defined as a strain having a 1/10(6) or greater heterogeneously resistant subpopulation to vancomycin . In another set of investigations, no hetero-VRSA were found among 12 other MRSA strains isolated after intravenous administration of vancomycin for 14 or more days (range: 14 to 77 days) . We conclude that while the use of intravenous vancomycin may have slightly lowered the vancomycin susceptibility of MRSA in our hospital, the decrease in so small that it may not be significant clinically . In addition, no hetero-VRSA were found in our hospial. Cutis, 2000 Dec, 66(6), 447 - 52 Septic embolization arising from infected pseudoaneurysms following percutaneous transluminal coronary angioplasty: a report of 2 cases and review of the literature; Izumi AK et al.; Septic embolization arising from infected pseudoaneurysms following percutaneous transluminal coronary angioplasty (PTCA) constitutes a distinct clinical and histopathologic entity . Pseudoaneurysms are a potential complication of both cardiac catheterization and PTCA . Repeated or prolonged catheterization increases the risk of bacterial seeding of these sites, resulting in septic embolization . Characteristic clinical features include fever within 2 to 5 days, unilateral embolic disease, and Staphylococcus aureus septicemia . Culture and examination of biopsy specimens of the embolic lesions typically demonstrate gram-positive microorganisms . We describe 2 patients presenting with ipsilateral palpable purpura, petechiae, and livedo reticularis caused by septic emboli from infected pseudoaneurysms . The recommended treatment includes administration of appropriate systemic antibiotics and surgical resection of the infected pseudoaneurysm . Both cholesterol and septic emboli should be considered in the differential diagnosis of ipsilateral embolic disease induced by invasive vascular procedures. Int J Antimicrob Agents, 2000 Nov, 16 Suppl 1, S39 - 42 Gram positive infection in trauma patients: new strategies to decrease emerging Gram-positive resistance and vancomycin toxicity; Ginzburg E et al.; Bacterial resistance to antibiotics has become a serious problem in medicine . Particularly worrisome is the increasing incidence of multi-resistant organisms such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) . Not surprisingly, in view of the high incidence of life-threatening infections and heavy antibiotic use, resistance has become very frequent and problematic in intensive care units . The standard approach for the treatment of MRSA is vancomycin or teicoplanin . Long-term therapeutic and unrestricted prophylactic use of vancomycin has given rise to VRE which in turn may lead to the emergence of vancomycin-resistant S . aureus (VRSA) through plasmid mediated transmission . In order to reduce the incidence of VRE and to avoid the emergence of VRSA, vancomycin use should be restricted and alternative antibiotic strategies should be developed . Using those antibiotics to which MRSA are still generally sensitive, perhaps in combination with new ones, such as, quinupristin/dalfopristin, should be entertained . We performed a retrospective review of the Gram-positive infections in our Level 1 Trauma Center Intensive Care Unit, and an analysis of the resistance patterns of the NMSA infections showed that additional resistance rarely develops within less than 5 days . We then designed a new strategy for the treatment of MRSA infections . This strategy consists of the sequential use of a range of antibiotics with activity against MRSA in short 5-7 day pulses until the full clinical course is completed . Studies validating the benefit of this approach are currently in preparation. Int J Antimicrob Agents, 2000 Nov, 16 Suppl 1, S31 - 4 Vancomycin treatment failures in Staphylococcus aureus lower respiratory tract infections; Moise PA et al.; We reviewed all patients with Staphylococcus aureus lower respiratory tract infections treated with vancomycin at our institution in 1998, to see how this antimicrobial is performing . We found that approximately 40% of evaluable patients were considered treatment failures, even though the S . aureus was still reported as being susceptible to vancomycin . We report in detail two example patients that failed to respond clinically to vancomycin and summarize the clinical characteristics of the 23 additional patients that failed . The first case was treated four times in the intensive care unit with vancomycin . Each course, after approximately 14 days therapy, the vancomycin was discontinued and his infection relapsed soon thereafter . The second was treated with vancomycin for 10 days initially . She relapsed, was restarted on vancomycin once more, but her condition deteriorated, and she died of refractory sepsis 20 days after admission . The cost of care for each patient ranged from $50,000 to over $100,000 . With trends such as these, alternative therapies are needed to control resistant Gram-positive infections. N Engl J Med, 2001 Jan 4, 344(1), 11 - 6 Nasal carriage as a source of Staphylococcus aureus bacteremia . Study Group; von Eiff C et al.; BACKGROUND: The consequences of infection with Staphylococcus aureus can be severe, so strategies for prevention are important . We examined S . aureus isolates from blood and from nasal specimens to determine whether the organisms in the bloodstream originated from the patient's own flora . METHODS: In a multicenter study, swabs for culture were obtained from the anterior nares of 219 patients with S . aureus bacteremia . A total of 723 isolates were collected and genotyped . In a second study, 1640 S . aureus isolates from nasal swabs were collected over a period of five years and then compared with isolates from the blood of patients who subsequently had S . aureus bacteremia . RESULTS: In the multicenter study of S . aureus bacteremia, the blood isolates were identical to those from the anterior nares in 180 of 219 patients (82.2 percent) . In the second study, 14 of 1278 patients who had nasal colonization with S . aureus subsequently had S . aureus bacteremia . In 12 of these 14 patients (86 percent), the isolates obtained from the nares were clonally identical to the isolates obtained from blood 1 day to 14 months later . CONCLUSIONS: A substantial proportion of cases of S . aureus bacteremia appear to be of endogenous origin since they originate from colonies in the nasal mucosa . These results provide support for strategies to prevent systemic S . aureus infections by eliminating nasal carriage of S . aureus. J Clin Microbiol, 2001 Jan, 39(1), 328 - 31 Binary typing of Staphylococcus aureus strains through reversed hybridization using digoxigenin-universal linkage system-labeled bacterial genomic DNA; van Leeuwen W et al.; A novel binary typing (BT) procedure, based on reversed hybridization of digoxigenin-universal linkage system-labeled bacterial DNA to strip-immobilized probes, is presented . Chromogenic detection of hybrids was performed . Staphylococcus aureus isolates (n = 20) were analyzed to establish the feasibility of BT . A technically simple and fast procedure has been developed for application in routine microbiology laboratories. J Clin Microbiol, 2001 Jan, 39(1), 86 - 9 International multicenter evaluation of latex agglutination tests for identification of Staphylococcus aureus; van Griethuysen A et al.; A newly marketed rapid agglutination kit for the identification of Staphylococcus aureus, Slidex Staph Plus (bioMerieux), was compared to Staphaurex Plus (Murex Diagnostics) and Pastorex Staph-Plus (Sanofi Diagnostics Pasteur) . The study took place in three clinical microbiology laboratories in three different European countries . A total of 892 staphylococcal isolates, including 278 methicillin-sensitive S . aureus (MSSA) isolates, 171 methicillin-resistant S . aureus (MRSA) isolates, and 443 coagulase-negative staphylococcal isolates, were analyzed . The sensitivities (MSSA/MRSA) and specificities, respectively, were 98 . 2% (98.9%/97.1%) and 98.9% for Slidex Staph Plus, 98.2% (98.2%/98 . 2%) and 96.2% for Staphaurex Plus, and 98.7% (98.6%/98.8%) and 95.7% for Pastorex Staph Plus . The specificity of the Slidex Staph Plus kit was statistically significantly higher than the specificities of Staphaurex Plus and Pastorex Staph-Plus . The Slidex Staph Plus is a very reliable test for the identification of S . aureus. J Clin Microbiol, 2001 Jan, 39(1), 53 - 6 Comparison of the Vitek Gram-Positive Susceptibility 106 card and the MRSA-screen latex agglutination test for determining oxacillin resistance in clinical bloodstream isolates of Staphylococcus aureus; Yamazumi T et al.; The Vitek automated susceptibility testing system with a modified Gram-Positive Susceptibility (GPS) 106 Card (bioMerieux Vitek, Inc., Hazelwood, Mo.) and a rapid slide latex agglutination test (MRSA-Screen; Denka Seiken Co., Ltd., Tokyo, Japan) were evaluated for their ability to detect oxacillin resistance in Staphylococcus aureus . The oxacillin-salt agar screen (OS) test, the reference broth microdilution method, and the detection of the mecA gene by PCR were compared with the commercial products . A total of 200 contemporary (1999) bloodstream infection isolates were collected from the SENTRY Antimicrobial Surveillance Program, representing diverse geographic areas throughout the world . Among the 99 mecA-positive isolates, 3 isolates were found negative by the MRSA-Screen . Another two isolates did not grow on OS plates and had MICs of 0.5 and 2 microg/ml with the Vitek GPS card . All 101 mecA-negative isolates were also found negative by the MRSA-Screen and were categorized as susceptible by the GPS card . Overall, the MRSA-Screen, GPS card, and OS test had sensitivities of 96.9, 98.0, and 98.0% and specificities of 100.0, 100.0, and 98.0%, respectively . MRSA-Screen was a rapid (</=15 min) and simple test to perform, and the GPS card provided results in <8 h . Both methods were sensitive and specific for detecting staphylococcal oxacillin resistance in the clinical microbiology laboratory. Braz J Infect Dis, 2000 Dec, 4(6), 296 - 300 The Effect of post-discharge surveillance and control strategies on the course of a Staphylococcus aureus outbreak in a newborn nursery; Couto RC et al.; This study was carried out to evaluate changes in infection rates following the adoption of three measures for controlling a Staphylococcus aureus outbreak in a nursery . In late April and early May, 1995, an outbreak of pustular dermatitis and conjunctivitis caused by S . aureus was documented . Case patients were identified by both in-hospital and post-discharge surveillance . In-hospital surveillance included daily review of data gathered by the hospital's infection control committee; use of microbiology laboratory results; requesting charts for antibiotic prescriptions; nurses' notes, and ward rounds surveillance as indicators of S . aureus infection (SAi) . Post-discharge surveillance was done by telephone survey around the 30th day after a baby's birth date . During the epidemic period, reinforcement of handwashing, daily bathing, and cord care with antiseptics combined with a cohort system of admissions, and nasal mupirocin ointment were introduced sequentially in an attempt to control the outbreak . The efficacy of these three strategies was measured through the decrease of SAi rates, detected by both in-hospital and post-discharge surveillance . A total of 5,639 babies were included in the study . In-hospital surveillance information was obtained from all patients and post-discharge surveillance in 3,506 (62.7%) patients . A total of 534 SAi were detected during the study, 47 in-hospital and 487 at post-discharge surveillance . A progressive decrease in the SAi rates could be observed after the institution of the control strategies, both in-hospital (7.6% to 0.5%) and after discharge (68.9% . to 11.7%) . During the study period, the rates of infection detected in out-patients were consistently higher than those diagnosed in-hospital . Improving handwashing and nasal mupirocin ointment seem to be more effective than cohort admissions and bath and cord care with antiseptics . The high SAi rates detected only after discharge from the hospital indicate that data from post-discharge surveillance should be included to estimate the true rates of infections in newborns. Nat Biotechnol, 2001 Jan, 19(1), 62 - 5 Interference-based detection of nucleic acid targets on optically coated silicon; Jenison R et al.; Sequence-specific detection of polynucleotides typically requires modified reporter probes that are labeled with radioactive, fluorescent, or luminescent moieties . Although these detection methods are capable of high sensitivity, they require instrumentation for signal detection . In certain settings, such as clinical point of care, instrumentation might be impractical or unavailable . Here we describe a detection approach in which formation of a nucleic acid hybrid is enzymatically transduced into a molecular thin film that can be visually detected in white light . The system exploits a flat, optically coated silicon-based surface to which capture oligonucleotides are covalently attached . The optimized system is capable of detection of nucleic acid targets present at sub-attomole levels . To supplement visual detection, signals can be quantitated by a charge-coupled device . The design and composition of the optical surface, optimization of immobilization chemistry for attachment of capture probes, and characterization of the efficiency of the hybridization process are presented . We describe the application of this system to detection of a clinically relevant target, the mecA gene present in methicillin-resistant Staphylococcus aureus. J Bacteriol, 2001 Jan, 183(2), 468 - 75 In Staphylococcus aureus, fur is an interactive regulator with PerR, contributes to virulence, and Is necessary for oxidative stress resistance through positive regulation of catalase and iron homeostasis; Horsburgh MJ et al.; The Staphylococcus aureus genome encodes three ferric uptake repressor (Fur) homologues: Fur, PerR, and Zur . To determine the exact role of Fur in S . aureus, we inactivated the fur gene by allelic replacement using a tetracycline resistance cassette, creating strain MJH010 (fur) . The mutant had a growth defect in rich medium, and this defect was exacerbated in metal-depleted CL medium . This growth defect was partially suppressed by manganous ion, a metal ion with known antioxidant properties . This suggests that the fur mutation leads to an oxidative stress condition . Indeed, MJH010 (fur) has reduced levels of catalase activity resulting from decreased katA transcription . Using a katA-lacZ fusion we have determined that Fur functions, either directly or indirectly, as an iron-dependent positive regulator of katA expression . Transcription of katA is coregulated by Fur and PerR, since in MJH010 (fur) transcription was still repressed by manganese while transcription in MJH201 (fur perR) was unresponsive to the presence of iron or manganese . Siderophore biosynthesis was repressed by iron in 8325-4 (wild-type) but in MJH010 (fur) was constitutive . A number of putative Fur-regulated genes were identified in the incomplete genome databases using known S . aureus Fur box sequences . Of those tested, the sstABCD and sirABC operons and the fhuD2 and orf4 genes were found to have Fur-regulated expression . MJH010 (fur) was attenuated (P<0.04) in a murine skin abscess model of infection, as was double-mutant MJH201 (fur perR) (P<0.03) . This demonstrates the importance in vivo of iron homeostasis and oxidative stress resistance regulation in S . aureus. Nat Biotechnol, 2001 Jan, 19(1), 66 - 70 Lysostaphin expression in mammary glands confers protection against staphylococcal infection in transgenic mice; Kerr DE et al.; Infection of the mammary gland, in addition to causing animal distress, is a major economic burden of the dairy industry . Staphylococcus aureus is the major contagious mastitis pathogen, accounting for approximately 15-30% of infections, and has proved difficult to control using standard management practices . As a first step toward enhancing mastitis resistance of dairy animals, we report the generation of transgenic mice that secrete a potent anti-staphylococcal protein into milk . The protein, lysostaphin, is a peptidoglycan hydrolase normally produced by Staphylococcus simulans . When the native form is secreted by transfected eukaryotic cells it becomes glycosylated and inactive . However, removal of two glycosylation motifs through engineering asparagine to glutamine codon substitutions enables secretion of Gln(125,232)-lysostaphin, a bioactive variant . Three lines of transgenic mice, in which the 5'-flanking region of the ovine beta-lactoglobulin gene directed the secretion of Gln(125,232)-lysostaphin into milk, exhibit substantial resistance to an intramammary challenge of 104 colony-forming units (c.f.u.) of S . aureus, with the highest expressing line being completely resistant . Milk protein content and profiles of transgenic and nontransgenic mice are similar . These results clearly demonstrate the potential of genetic engineering to combat the most prevalent disease of dairy cattle. J Biol Chem, 2001 Mar 23, 276(12), 8875 - 83 Epub 2000 Dec 21. Significant interference with hepatitis B virus replication by a core-nuclease fusion protein; Beterams G et al.; Hepatitis B virus (HBV), a small DNA containing virus that replicates via reverse transcription, causes acute and chronic B-type hepatitis in humans . The limited success of current therapies for chronic infection has prompted exploration of alternative strategies . Capsid-targeted viral inactivation is a conceptually powerful approach that exploits virion structural proteins to target a degradative enzyme specifically into viral particles . Its principal feasibility has been demonstrated in retroviral model systems but not yet for a medically relevant virus outside the retrovirus family . Recently, we found that C proximal fusion to the HBV capsid protein of the Ca(2+)-dependent nuclease (SN) from Staphylococcus aureus yields a chimeric protein, coreSN, that in Escherichia coli coassembles with the wild-type capsid protein into particles with internal SN domains . Here we show that, in HBV co-transfected human hepatoma cells, less than 1 coreSN protein per 10 wild-type core protein subunits reduced titers of enveloped DNA containing virions by more than 95% . The antiviral effect depends on both an enzymatically active SN and on the core domain . CoreSN does not block assembly of RNA containing nucleocapsids but interferes with proper synthesis of viral DNA inside the capsid, or leads to rapid DNA degradation . Our data suggest an intracellular nuclease activation that, owing to the characteristics of HBV morphogenesis, is nonetheless highly virus specific . HBV may therefore be particularly vulnerable to the capsid-targeted viral inactivation approach. J Antibiot (Tokyo), 2000 Oct, 53(10), 1045 - 52 SAR studies of anti-MRSA non-zwitterionic 3-heteroarylthiocephems; Glinka TW et al.; SAR studies in a series of 3-heteroarylthio substituted cephalosporins established that high activity against methicillin-resistant Staphylococcus aureus (MRSA) can be achieved with various heteroaryl substituents . Early results showed that highly lipophilic 3-heteroarylthio substituents, which were necessary for anti-MRSA activity, caused high affinity of such cephems toward serum proteins . Our earlier published efforts described discovery of zwitterionic cephems MC-02,331 and RWJ-54428 (MC-02,479), where serum binding was reduced by employing basic, positively charged functionalities attached to the 3-heteroarylthio substituent . In order to avoid low solubility problems associated with most such zwitterionic cephalosporins a wide variety of non-basic heteroaryl substituents was tested (non-zwitterionic cephems are more easily formulated as water soluble sodium salts for intravenous administration) . Considerable reduction in serum binding was obtained in some analogs while maintaining high anti-MRSA potency. J Antibiot (Tokyo), 2000 Oct, 53(10), 1028 - 37 Cephems: fifty years of continuous research; Bryskier A; Since 1964, seven waves of parenteral cephems have been reported . All of them were designed to meet medical needs . The first (group I) and the third (group III) waves were very successful and drugs belonging to group III are widely used in the treatment of severe infections . A new series of compounds (group VII), with a new compound underdevelopment was designed for the treatment of Staphylococcus aureus strain resistant to methicillin but also to glyco- and lipoglycopeptides . By modifying the substituent at position 3 and 7 of the cephems rings optimal moieties have been fixed leading to potent anti-Gram-positive drugs . Alterations of substituents are still in progress to obtain optimal anti-Gram-positive (anti-MRSA) compounds . The first oral cephem cephalexin was introduced in clinical practice in 1967 . Since this time, many esterified and non-esterified cephems have been synthesized and introduced in clinics . There are two groups of compounds, alpha-amino and non-alpha-amino cephems which are classified in six groups according to their chemical structure . The absorption route was explored and three transporting systems have been described according to the physico-chemical properties of these compounds, in addition prodrugs are passively absorbed. Trans R Soc Trop Med Hyg, 2000 Sep-Oct, 94(5), 504 - 7 Nasal carriage and antibiotic resistance of Staphylococcus aureus isolates from hospital and non-hospital personnel in Abha, Saudi Arabia; Alghaithy AA et al.; The prevalence of nasal carriage of Staphylococcus aureus, antibiograms and prevalence of methicillin-resistant S . aureus (MRSA) were studied in 1999 among healthy hospital and non-hospital personnel in Abha, Saudi Arabia . S . aureus was isolated from 26.1% of 299 adults in the community and 25.4% of 279 hospital personnel . No isolate was resistant to vancomycin . Antibiotic resistance rates, for all other antibiotics tested except cephalothin, were significantly higher for strains from hospital personnel (P values < 0.001-0.04) compared to non-hospital adults . The antibiograms were also compared with those of 140 clinical isolates . The rates of resistance of the inpatient strains to all the antibiotics tested were significantly higher than those of hospital nasal carrier strains (P < 0.001-0.05) . MRSA was isolated, respectively, from 5.1% and 18.3% of non-hospital and hospital carriers; MRSA carriage rates were 1.3% and 4.7%, respectively, for non-hospital and hospital carriers, and 61% of S . aureus isolates from infected patients were MRSA . Only 8% of non-hospital but 44% of hospital carrier strains were multiply resistant (P < 0.001) . Multiple resistance among inpatient strains (89%) was significantly higher than that among hospital nasal strains (44%) (P < 0.001) . Such rates of multiple resistance and endemic MRSA prevalence among healthy carriers (11%) at a much higher rate than those reported in the literature should raise concern in a region with unrestricted availability of antibiotics. J Chemother, 2000 Nov, 12 Suppl 5, 40 - 55 Does surgical prophylaxis with teicoplanin constitute a therapeutic advance? Mini E, Nobili S, Periti P. Antibiotic prophylaxis has become standard care not only in operations characterized by high infection rates but also in the vast majority of clean surgical procedures, including those that use foreign materials, grafts or prosthetic devices as well as non-implant surgery . While use of antibiotics in clean implant surgery is undisputed, it is still controversial in clean non-implant surgery . As antibiotic prophylaxis should be directed against expected pathogens, the glycopeptides are considered suitable alternative antibiotics to first and second generation cephalosporins in clean surgical procedures associated with a high risk of wound infections due to Gram-positive bacteria, including methicillin-resistant, and for patients allergic to beta-lactam antibiotics . In deciding whether to use a glycopeptide for prophylaxis, the current wound infection rates with methicillin-resistant Staphylococcus aureus and methicillin-resistant Staphylococcus epidermidis at single institutions need to be considered, to limit the use of glycopeptides to wards where the incidence of methicillin resistance is high . Of the two available glycopeptides, teicoplanin may be preferable to vancomycin for peri-operative prophylaxis because of its excellent tissue penetration, as indicated by the large volume of distribution, lower toxicity, and particularly long half-life, allowing single-dose administration in several surgical procedures . Clinical trials with teicoplanin prophylaxis in several types of clean surgical procedures including orthopedic, cardiac, vascular and dental operations, have shown it to be efficacious . This review focuses on results from clinical studies with this glycopeptide as prophylaxis in clean surgery. J Chemother, 2000 Nov, 12 Suppl 5, 26 - 33 Teicoplanin in the treatment of serious infection; Schaison G et al.; The merits and dosing regimens for teicoplanin in the treatment of endocarditis, bacteremia, bone and joint infections, pediatric use, non in-patient use and in the ICU are discussed . Teicoplanin has several advantages over vancomycin in the treatment of serious infections: long half-life, lower nephrotoxicity, and lack of requirement for serum assays . The recommended regimen for teicoplanin is three loading doses of 6 mg/kg (400 mg) q12h, then 6 mg/kg (400 mg) q24h . There is no significant difference in efficacy between teicoplanin and vancomycin when at least 6 mg/kg teicoplanin is used or, in the case of staphylococcal endocarditis, it is given in combination with another antimicrobial . Teicoplanin is effective and safe in staphylococcal infections including endocarditis, osteomyelitis and septic arthritis . The once daily or alternate daily dosage allows home administration of treatment of infections caused by Staphylococcus aureus, including methicillin-resistant strains and enterococci with appreciable savings in hospital costs and improvement in the quality of life. Vet Res, 2000 Nov-Dec, 31(6), 603 - 9 Decreased neutrophil bactericidal activity during phagocytosis of a slime-producing Staphylococcus aureus strain; Barrio B et al.; Phagocytosis and intracellular killing by bovine polymorphonuclear leukocytes (PMN) are important host defence mechanisms against mastitis caused by Staphlylococcus aureus . We compared the phagocytosis and overall killing of a non slime-producing (NSP) S . aureus and its slime-producing (SP) variant by blood PMN, using an in vitro bacteriological assay . Seven clinically healthy Holstein-Friesian dairy cows in mid-lactation stage were used for this purpose . The percentages of overall killing for the NSP and SP variant were 34+/-3% and 21+/-4% (P < 0.05) and the corresponding percentages of phagocytosis were 40+/-4% and 31+/-4%, respectively . A significant positive correlation (r = 0.79; P < 0.001) was found between phagocytosis and overall killing . These results suggest that the presence of slime was responsible for a decreased phagocytic ingestion and overall killing. J Med Microbiol, 2000 Dec, 49(12), 1109 - 17 Contamination of expressed human breast milk with an epidemic multiresistant Staphylococcus aureus clone; Novak FR et al.; Nosocomial infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are a major cause of outbreaks in intensive care units . Infants make up a sector of the population that presents a high risk for MRSA infections . Mother-to-infant transmission has been indicated as a possible cause of MRSA infections in neonates . The occurrence and characteristics of MRSA in samples of banked human milk were investigated by selective culture, antibiogram and pulsed-field gel electrophoresis . MRSA contamination was found in 11% of 500 samples of expressed, fresh-frozen milk from 500 different donors at five Brazilian milk banks . The great majority of the contaminated samples passed breast milk quality control criteria for dispensing as raw milk under Brazilian and American guidelines . Most of the MRSA isolates belonged to the Brazilian epidemic clone, which is reported to be widespread in several Brazilian states, in Argentina and in Portugal . It is concluded that expressed breast milk can be a reservoir of multiresistant S . aureus epidemic clones . Studies are necessary to assess the source of contamination and potential role of MRSA-contaminated milk in the transmission of MRSA to neonates. J Med Microbiol, 2000 Dec, 49(12), 1103 - 7 Co-transfer of plasmids in association with conjugative transfer of mupirocin or mupirocin and penicillin resistance in methicillin-resistant Staphylococcus aureus; Pawa A et al.; Two distinct strains of methicillin-resistant Staphylococcus aureus (MRSA) isolated from patients in a dermatology ward were also resistant to mupirocin . The mupirocin resistance plasmids from both strains were indistinguishable by EcoRI and HindIII restriction digest analysis, except for the presence of genes apparently mediating penicillinase production in some transconjugants . Conjugative transfer of the plasmid mediating mupirocin resistance from one of these strains to a recipient S . aureus was accompanied in some cases by co-transfer of plasmids mediating resistance to tetracycline or erythromycin; in some instances a plasmid which possessed no apparent resistance markers was also transferred . The second strain demonstrated conjugative transfer of penicillin and mupirocin resistance as well as transfer of a plasmid mediating gentamicin resistance, but transfer of erythromycin resistance was not apparently plasmid-mediated. Laryngoscope, 2000 Dec, 110(12), 2085 - 8 Effects of beta-toxin of Staphylococcus aureus on ciliary activity of nasal epithelial cells; Kim CS et al.; OBJECTIVES: To investigate the in vitro effects of staphylococcal beta-toxin on ciliary activity and the in vivo effects on sinusitis induction . STUDY DESIGN: The in vitro effects of staphylococcal beta-toxin on ciliary activity were investigated at different concentrations and exposure times . Experimental sinusitis was induced in rabbits with application of beta-toxin and confirmed 7 days later . METHODS: Ciliated epithelial cells were taken from the maxillary sinus mucosa of 10 rabbits . Five culture dishes from each rabbit were used for the experimental group, and one culture dish from each rabbit was used for the control group . In the experimental group, ciliary beat frequency (CBF) was measured at concentrations of 0.1, 1, 2, 5 and 10 U/mL of beta-toxin using a video-computerized analysis technique, while in the control group, culture medium containing no toxin was used . CBF was measured 1, 2, 4, 6, 8, 12, 24, and 48 hours after administration of beta-toxin . To induce experimental sinusitis, 2 U/mL of beta-toxin was percutaneously applied to the maxillary sinus of 10 rabbits without occlusion of the natural ostium, while normal saline was percutaneously applied to the right-side maxillary sinus of 4 rabbits in the control group . At 7 days, mucosal membranes were taken from the inferomedial wall of the maxillary sinus for light microscopic study . RESULTS: CBF dropped significantly after an 8-hour incubation at 2, 5, and 10 U/mL of beta-toxin . No ciliary activity was observed after a 24-hour incubation at 2 and 5 U/mL and a 12-hour incubation at 10 U/mL of beta-toxin . Mucoid, purulent discharge was observed in the maxillary sinuses of the beta-toxin-applied group . Prominent epithelial disruption and infiltration of inflammatory cells into the epithelium and lamina propria were observed in the beta-toxin-applied group . CONCLUSIONS: Staphylococcal beta-toxin may reduce ciliary activity and induce sinusitis without occlusion of the natural ostium of the maxillary sinus in rabbits This study provides another animal model of sinusitis for understanding the pathogenesis of sinusitis induced by bacterial exotoxins. Anal Chem, 2000 Dec 1, 72(23), 5761 - 5 Immunoassay of the MRSA-related toxic protein, leukocidin, with scanning electrochemical microscopy; Kasai S et al.; Scanning electrochemical microscopy (SECM) was applied to the immunoassay of leukocidin, which is a toxic protein produced by methicillin-resistant Staphylococcus aureus (MRSA), with the intention of developing and early diagnostic for MRSA infection . An antibody-chip for leukocidin was prepared by self-assembling of anti-leukocidin on a protein A-coated glass substrate . A sample solution containing leukocidin was spotted onto the antibody-chip, followed by labeling with horseradish peroxidase (HRP) via a sandwich method . The reduction current of the oxidized form of ferrocenylmethanol generated by the HRP reaction was monitored to view SECM images of the spot of captured leukocidin . The amplitude of reduction current depended on the concentrations of sample solutions used for making spots . This SECM-based immunoassay detects as low as 5.25 pg mL(-1) leukocidin. Bioorg Med Chem Lett, 2000 Dec 4, 10(23), 2675 - 8 Potent in vitro methicillin-resistant Staphylococcus aureus activity of 2-(1H-indol-3-yl)quinoline derivatives; Hoemann MZ et al.; A novel structural class of antibacterials, 2-(1H-indol-3-yl)quinolines, effective against methicillin-resistant Staphylococcus aureus (MRSA), was discovered from a combinatorial library . A structure-activity relationship (SAR) study was conducted to determine the pharmacophore and increase the potency of these compounds . Compounds were prepared that had minimum inhibitory concentrations (MICs) < 1.0 microg/mL against MRSA and retained activity against two strains of glycopeptide intermediate-resistant S . aureus (GISA). Indian J Pathol Microbiol, 1999 Oct, 42(4), 441 - 6 Nasal carriage of methicillin resistant Staphylococcus aureus in a cardiovascular tertiary care centre and its detection by Lipovitellin Salt Mannitol Agar; Verghese S et al.; Ecological niches of Staphylococcus aureus are the anterior nares . Carriage of Staphylococcus aureus in the nose appears to play a key role in the epidemiology and pathogenesis of infection . Numerous studier have shown that elimination of nasal carriage using Mupirocin also eliminated hand carriage and the spread of infections in hospitals . Lipovitellin-Salt-Mannitol Agar was used for screening, isolation and presumptive identification of Staphylococcus aureus from nasal carriers . From November; 97 to August'98, 724 nasal swabs were cultured and 18.23% of health care workers were found to be nasal carriers of Staphylococcus aureus . Of these 12.15% were carriers of MRSA . The carrier rate was highest in December' 97 (32.07%) . All MRSA carriers were treated with local application of Mupirocin for three days . A study of the antibiogram of the clinical isolates during the corresponding period showed 100% susceptibility of MRSA to Vancomycin . Susceptibility of MRSA to Clindamycin, Netilmycin, Rifampicin & Ofloxacin was 86.6%, 69.5%, 66% & 64.7% respectively. Indian J Pathol Microbiol, 1999 Oct, 42(4), 421 - 6 Community acquired methicillin resistant Staphylococcsus aureus: a new threat for hospital outbreaks? Gupta N, Prakash SK, Malik VK, Mehndiratta PL, Mathur MD. Methicillin resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen . Recently, there have been reports of increasing prevalence of MRSA in the community . We here report an outbreak of post operative wound sepsis by MRSA in the surgical ward of LN hospital . A surveillance study for MRSA was undertaken in the corresponding surgical ward, operation theater and OPD and the source of this outbreak was traced to an outdoor patient with community acquired MRSA infection . A total of 320 clinical and environmental samples were screened for MRSA . Seventy (21.8%) S . aureus were obtained, of which 12.8% were resistant to methicillin . 14% of the MRSA infections were from the community . Nasal carriage rates of MRSA in the screened hospital staff and admitted patients were 5.8% and 4.3% respectively . None of the environmental sites sampled yielded MRSA . A study of antibiogram revealed that all the MRSA were uniformly resistant to penicillin, erythromycin, gentamicin, tobramycin and tetracycline and sensitive to vancomycin . All isolates belonged to the same biotype and were nontypable by the standard set of phages. J Laryngol Otol, 2000 Oct, 114(10), 796 - 7 Mycotic pseudoaneurysm of common carotid artery mimicking parapharyngeal abscess; Constantinides H et al.; In this case a secondarily infected pseudoaneurysm of the common carotid artery presented with clinical features suggestive of a parapharyngeal abscess . The causative organism was identified as community-acquired methicillin-resistant Staphylococcus aureus . To the authors' knowledge this condition not previously been reported. Eur J Dermatol, 2000 Dec, 10(8), 630 - 2 Staphylococcal scalded skin syndrome with prosthetic valve endocarditis; Ansai SI et al.; We report staphylococcal scalded skin syndrome (SSSS) in a 67-year-old man . He showed diffuse erythema with erosion on his face and erythema with giant desquamation on his neck, axilla, genitalia, chest and abdomen 39 days after a coronary artery bypass graft and aortic valve replacement . He died of cardiac rupture caused by myocardial necrosis, and autopsy findings demonstrated prosthetic valve endocarditis due to a strain of exfoliative toxin-B producing methicillin-resistant Staphylococcus aureus . To the best of our knowledge, this is the first case of SSSS caused by prosthetic valve endocarditis. Arch Dis Child Fetal Neonatal Ed, 2001 Jan, 84(1), F38 - 9 Acquired subglottic stenosis caused by methicillin resistant Staphylococcus aureus that produce epidermal cell differentiation inhibitor; Yamada Y et al.; Local infection of the trachea in intubated neonates is one of the main risk factors for development of acquired subglottic stenosis, although its role in the pathogenesis is unclear . Methicillin resistant Staphylococcus aureus (MRSA) is often the cause of critical illness in neonatal patients . Two cases are reported of acquired subglottic stenosis following bacterial infection of the trachea, suggesting an association with the staphylococcal exotoxin, epidermal cell differentiation inhibitor (EDIN) . EDIN-producing MRSA were isolated from purulent tracheal secretions from both infants . Acquired subglottic stenosis in both cases was probably caused by delayed wound healing as the result of EDIN inhibition of epithelial cell migration. J Immunol, 2001 Jan 1, 166(1), 669 - 77 egc, a highly prevalent operon of enterotoxin gene, forms a putative nursery of superantigens in Staphylococcus aureus; Jarraud S et al.; The recently described staphylococcal enterotoxins (SE) G and I were originally identified in two separate strains of Staphylococcus aureus . We have previously shown that the corresponding genes seg and sei are present in S . aureus in tandem orientation, on a 3.2-kb DNA fragment (Jarraud, J . et al . 1999 . J . Clin . Microbiol . 37:2446-2449) . Sequence analysis of seg-sei intergenic DNA and flanking regions revealed three enterotoxin-like open reading frames related to seg and sei, designated sek, sel, and sem, and two pseudogenes, psi ent1 and psi ent2 . RT-PCR analysis showed that all these genes, including seg and sei, belong to an operon, designated the enterotoxin gene cluster (egc) . Recombinant SEG, SEI, SEK, SEL, and SEM showed superantigen activity, each with a specific V beta pattern . Distribution studies of genes encoding superantigens in clinical S . aureus isolates showed that most strains harbored such genes and in particular the enterotoxin gene cluster, whatever the disease they caused . Phylogenetic analysis of enterotoxin genes indicated that they all potentially derived from this cluster, identifying egc as a putative nursery of enterotoxin genes. Antimicrob Agents Chemother, 2001 Jan, 45(1), 349 - 52 Clinical isolate of vancomycin-heterointermediate Staphylococcus aureus susceptible to methicillin and in vitro selection of a vancomycin-resistant derivative; Bobin-Dubreux S et al.; A Staphylococcus aureus strain with low-level heteroresistance to vancomycin (designated MER) but susceptible to methicillin was isolated from an outpatient with conjunctivitis who did not receive any glycopeptide antibiotics . Incubation of the parent strain, MER, with increasing concentrations of vancomycin led to rapid selection of a stable progeny homogeneously resistant to vancomycin . Electron micrographs of strain MER showed enhanced cell wall thickness and abnormal septations typically seen with methicillin-resistant S . aureus having intermediate susceptibility to vancomycin. Antimicrob Agents Chemother, 2001 Jan, 45(1), 280 - 7 A spectrum of changes occurs in peptidoglycan composition of glycopeptide-intermediate clinical Staphylococcus aureus isolates; Boyle-Vavra S et al.; The mechanism of glycopeptide resistance in Staphylococcus aureus is not known with certainty . Because the target of vancomycin is the D-Ala-D-Ala terminus of the stem peptide of the peptidoglycan precursor, by subjecting muropeptides to reversed-phase high-performance liquid chromatography, we investigated peptidoglycan obtained from glycopeptide-intermediate S . aureus (GISA) isolates for changes in composition and evaluated whether any peptidoglycan structural change was a consistent feature of clinical GISA isolates . GISA isolates Mu50 and Mu3 from Japan had the large glutamate-containing monomeric peak demonstrated previously, although strain H1, a vancomycin-susceptible MRSA isolate from Japan that was clonally related to Mu3 and Mu50, and a femC mutant that we studied, did also . For the U.S . GISA isolates, strain NJ had a large monomeric peak with a retention time identical to that described for the glutamate-containing monomer in strains H1, Mu3, and Mu50 . However, a much smaller corresponding peak was seen in GISA MI, and this peak was absent from both GISA PC and a recent GISA isolate obtained from an adult patient in Illinois (strain IL) . These data suggest that a uniform alteration in peptidoglycan composition cannot be discerned among the GISA isolates and indicate that a single genetic or biochemical change is unlikely to account for the glycopeptide resistance phenotype in the clinical GISA isolates observed to date . Furthermore, a large monomeric glutamate-containing peak is not sufficient to confer the resistance phenotype. Antimicrob Agents Chemother, 2001 Jan, 45(1), 208 - 11 In vivo activity of evernimicin (SCH 27899) against methicillin-resistant Staphylococcus aureus in experimental infective endocarditis; Boucher HW et al.; Currently, there exist few satisfactory alternatives to vancomycin for therapy of serious methicillin-resistant Staphylococcus aureus (MRSA) infections . We employed a rat model of aortic valve endocarditis to assess the potential efficacy of evernimicin (SCH 27899) compared with vancomycin against infection with a strain susceptible to both agents (MICs of 0.25 and 0.50 microg/ml, respectively) . Infected animals were assigned to one of three groups: controls (no treatment), evernimicin at 60 mg/kg of body weight by intravenous (i.v.) infusion once daily, or vancomycin at 150 mg/kg of body weight per day by continuous i.v . infusion . Therapy was administered for 5.5 days . At the start of therapy, colony counts in vegetations were 6.63 +/- 0.44 log(10) CFU/g . In both treatment groups, bacterial density within vegetations was significantly reduced in comparison with control animals that had not been treated . Final colony counts were as follows (mean +/- standard deviation): controls, 10.12 +/- 1.51 log(10) CFU/g of vegetation; evernimicin, 7.22 +/- 2.91 log(10) CFU/g of vegetation; vancomycin, 5.65 +/- 1.76 log(10) CFU/g of vegetation . The difference between the evernimicin and vancomycin groups was not significant . These results confirmed the bacteriostatic activity of evernimicin in vivo in an experimental model of severe MRSA infection. Antimicrob Agents Chemother, 2001 Jan, 45(1), 196 - 202 Effects of amoxicillin, gentamicin, and moxifloxacin on the hemolytic activity of Staphylococcus aureus in vitro and in vivo; Worlitzsch D et al.; In Staphylococcus aureus infection hemolysis caused by the extracellular protein alpha-toxin encoded by hla is thought to contribute significantly to its multifactorial virulence . In vitro, subinhibitory concentrations of beta-lactam antibiotics and fluoroquinolones increase the levels of hla and alpha-toxin expression, whereas aminoglycosides decrease the levels of hla and alpha-toxin expression . In the present study we investigated the effects of subinhibitory concentrations of amoxicillin, gentamicin, and moxifloxacin on hla and alpha-toxin expression and total hemolysis of S . aureus strain 8325-4, a high-level alpha-toxin producer, and its alpha-toxin-negative mutant, DU 1090, in vitro and in a rat model of chronic S . aureus infection . The levels of expression of hla and alpha-toxin and total hemolysis did not differ significantly when amoxicillin, gentamicin, or moxifloxacin was added to cultures of S . aureus strain 8325-4 . In vivo, strain 8325-4 induced a significantly increased level of hemolysis in infected pouches compared to that in uninfected control pouches, but the hemolysis was reduced to control levels by treatment with doses of amoxicillin, gentamicin, or moxifloxacin that reduced bacterial numbers by 2 orders of magnitude . Additionally, the effects of subinhibitory concentrations of the three antibiotics on total hemolysis of four methicillin-resistant S . aureus and three methicillin-sensitive S . aureus (MSSA) clinical isolates were assessed in vitro . A significant increase in total hemolysis was observed for only one MSSA strain when it was treated with amoxicillin but not when it was treated with moxifloxacin or gentamicin . When purified alpha-toxin was incubated with purified human neutrophil elastase, alpha-toxin was cleaved nearly completely . The results suggest that the penicillin-induced increases in S . aureus alpha-toxin expression are strain dependent, that reduction of bacterial numbers in vivo counteracts this phenomenon effectively, and finally, that in localized S . aureus infections alpha-toxin activity is controlled by neutrophil elastase. Antimicrob Agents Chemother, 2001 Jan, 45(1), 79 - 83 Effects of mutations in ribosomal protein L16 on susceptibility and accumulation of evernimicin; McNicholas PM et al.; Chemical mutagenesis of Staphylococcus aureus RN450 generated two strains that displayed a stable reduction (30- to 60-fold) in susceptibility to evernimicin . Cell-free translation reactions demonstrated that the resistance determinant was located in the ribosomal fraction . Compared to ribosomes isolated from a wild-type strain, ribosomes from the mutant strains displayed an 8- to 10-fold reduction in affinity for {(14)C}evernimicin . In contrast, the mutants displayed no alteration in either binding affinity or in vitro susceptibility to erythromycin . Exponential cultures of the mutant strains accumulated significantly less {(14)C}evernimicin than the wild-type strain, suggesting that accumulation is dependent on the high affinity that evernimicin displays for its binding site . Sequencing rplP (encodes ribosomal protein L16) in the mutant strains revealed a single base change in each strain, which resulted in a substitution of either cysteine or histidine for arginine at residue 51 . Introduction of a multicopy plasmid carrying wild-type rplP into the mutant strains restored sensitivity to evernimicin, confirming that the alterations in rplP were responsible for the change in susceptibility . Overexpression of the mutant alleles in S . aureus RN450 had no effect on susceptibility to evernimicin, demonstrating that susceptibility is dominant over resistance. J Immunol, 2000 Dec 15, 165(12), 6880 - 8 Expression and function of IL-12 and IL-18 receptors on human tonsillar B cells; Airoldi I et al.; IL-12 activates murine and human B cells, but little information is available as to the expression and function of IL-12R on human B lymphocytes . Here we show that the latter cells, freshly isolated from human tonsils, expressed the transcripts of both beta1 and beta2 chains of IL-12R and that beta2 chain mRNA was selectively increased (4- to 5-fold) by incubation with Staphylococcus aureus Cowan I bacteria or IL-12 . B cell stimulation with IL-12 induced de novo expression of the transcripts of the two chains of IL-18R, i.e., IL-1 receptor-related protein and accessory protein-like . Functional studies showed that both IL-12 and IL-18 signaled to B cells through the NF-kappaB pathway . In the case of IL-12, no involvement of STAT transcription factors, and in particular of STAT-4, was detected . c-rel and p50 were identified as the members of NF-kappaB family involved in IL-12-mediated signal transduction to B cells . IL-12 and IL-18 synergized in the induction of IFN-gamma production by tonsillar B cells, but not in the stimulation of B cell differentiation, although either cytokine promoted IgM secretion in culture supernatants . Finally, naive but not germinal center or memory, tonsillar B cells were identified as the exclusive IL-12 targets in terms of induction of NF-kappaB activation and of IFN-gamma production. Infect Immun, 2001 Jan, 69(1), 360 - 6 Biochemical and biological properties of Staphylococcal enterotoxin K; Orwin PM et al.; Staphylococcus aureus is an important human pathogen which is implicated in a wide variety of diseases . Major determinants of the virulence of this organism include extracellular virulence factors . Staphylococcal enterotoxins (SEs) are important causative agents in staphylococcal toxic shock syndrome and food poisoning . Our study identified a novel enterotoxin, SEK, and examined its biochemical and biological properties . SEK had a molecular weight of 26,000 and an experimentally determined pI of between 7.0 and 7.5 . SEK was secreted by clinical isolates of S . aureus . We demonstrated that SEK had many of the biological activities associated with the SEs, including superantigenicity, pyrogenicity, the ability to enhance the lethal effect of endotoxin, and lethality in a rabbit model when administered by subcutaneous miniosmotic pump . Recombinant SEK was shown to stimulate human CD4(+) and CD8(+) T cells in a Vbeta-specific manner; T-cells bearing Vbeta 5.1, 5.2, and 6.7 were significantly stimulated to proliferate. Infect Immun, 2001 Jan, 69(1), 345 - 52 Clonal associations among Staphylococcus aureus isolates from various sites of infection; Booth MC et al.; A molecular epidemiological analysis was undertaken to identify lineages of Staphylococcus aureus that may be disproportionately associated with infection . Pulsed-field gel electrophoresis analysis of 405 S . aureus clinical isolates collected from various infection types and geographic locations was performed . Five distinct S . aureus lineages (SALs 1, 2, 4, 5, and 6) were identified, which accounted for 19.01, 9.14, 22.72, 10.12, and 4.69% of isolates, respectively . In addition, 85 lineages which occurred with frequencies of <2.5% were identified and were termed "sporadic." The most prevalent lineage was methicillin-resistant S . aureus (SAL 4) . The second most prevalent lineage, SAL 1, was also isolated at a high frequency from the anterior nares of healthy volunteers, suggesting that its prevalence among clinical isolates may be a consequence of high carriage rates in humans . Gene-specific PCR was carried out to detect genes for a number of staphylococcal virulence traits . tst and cna were found to be significantly associated with prevalent lineages compared to sporadic lineages . When specific infection sites were examined, SAL 4 was significantly associated with respiratory tract infection, while SAL 2 was enriched among blood isolates . SAL 1 and SAL 5 were clonally related to SALs shown by others to be widespread in the clinical isolate population . We conclude from this study that at least five phylogenetic lineages of S . aureus are highly prevalent and widely distributed among clinical isolates . The traits that confer on these lineages a propensity to infect may suggest novel approaches to antistaphylococcal therapy. Infect Immun, 2001 Jan, 69(1), 159 - 69 Description of staphylococcus serine protease (ssp) operon in Staphylococcus aureus and nonpolar inactivation of sspA-encoded serine protease; Rice K et al.; Signature tagged mutagenesis has recently revealed that the Ssp serine protease (V8 protease) contributes to in vivo growth and survival of Staphylococcus aureus in different infection models, and our previous work indicated that Ssp could play a role in controlling microbial adhesion . In this study, we describe an operon structure within the ssp locus of S . aureus RN6390 . The ssp gene encoding V8 protease is designated as sspA, and is followed by sspB, which encodes a 40.6-kDa cysteine protease, and sspC, which encodes a 12.9-kDa protein of unknown function . S . aureus SP6391 is an isogenic derivative of RN6390, in which specific loss of SspA function was achieved through a nonpolar allelic replacement mutation . In addition to losing SspA, the culture supernatant of SP6391 showed a loss of 22- to 23-kDa proteins and the appearance of a 40-kDa protein corresponding to SspB . Although the 40-kDa SspB protein could degrade denatured collagen, our data establish that this is a precursor form which is normally processed by SspA to form a mature cysteine protease . Culture supernatant of SP6391 also showed a new 42-kDa glucosaminidase and enhanced glucosaminidase activity in the 29 to 32 kDa range . Although nonpolar inactivation of sspA exerted a pleiotropic effect, S . aureus SP6391 exhibited enhanced virulence in a tissue abscess infection model relative to RN6390 . Therefore, we conclude that SspA is required for maturation of SspB and plays a role in controlling autolytic activity but does not by itself exert a significant contribution to the development of tissue abscess infections. Infect Immun, 2001 Jan, 69(1), 45 - 51 Staphylococcus aureus agr genotypes with enterotoxin production capabilities can resist neutrophil bactericidal activity; Mullarky IK et al.; Staphylococcus aureus pathogenicity is mainly due to the production of a number of secreted and cell surface-associated proteins under the regulation of the agr gene . A region of the agr gene was used to subgroup S . aureus strains according to restriction fragment length polymorphisms . Additionally, strains were subtyped according to the coagulase gene in order to strengthen discriminatory power . Virulence capabilities of agr genotype subgroups were evaluated using an in vitro neutrophil bactericidal assay, which showed that prevalent genotypes were significantly better at evading this primary host defense . Multiplex PCR was then used to detect enterotoxin genes among the genotype subgroups in order to determine possible virulence candidates that enable strains to combat neutrophil killing . The prevalent genotype strains were found to possess higher production capabilities for enterotoxin A than did low-prevalence strains . The significance of enterotoxin A production capabilities in affecting pathogenicity of S . aureus strains was evaluated and found to have a profound effect on neutrophil killing abilities . The use of a large epidemiological database as a tool for subgrouping strains with varying degrees of pathogenicity has allowed the identification of relevant and previously undefined virulence factors that affect a pathogen's capability to overcome host immune defenses. FEMS Immunol Med Microbiol, 2000 Dec, 29(4), 315 - 21 Identification of the secreted macromolecular immunogens of Staphylococcus aureus by analysis of serum; Royan S et al.; The ability of sera to recognise secreted macromolecules of Staphylococcus aureus was examined by ELISA and Western immunoblotting . Individual secreted proteins were also studied using both human sera and sera from rabbits immunised with secreted macromolecules . Patients sera showed a wide range of IgG antibody titres to secreted macromolecules and whole bacteria . Controls showed a significantly lower IgG response . Western immunoblotting revealed that a significant number of secreted proteins were recognised by circulating IgG antibodies . Surprisingly, both the sera from controls and from patients recognised similar macromolecules including a number of potential virulence factors . The major difference was in the IgG binding to a 16-kDa component, which was recognised by the majority of the sera from infected individuals, but only by a small number of sera from healthy controls . The higher incidence of antibodies recognising the 16 kDa component may be related to our earlier finding that the major bone resorbing component of S . aureus is a heterodimeric protein containing a 16-kDa subunit, the activity of which could be blocked by sera. Vet Microbiol, 2001 Jan 5, 78(1), 39 - 48 Genetic analysis of exfoliative toxin A-producing Staphylococcus aureus isolated from mastitic cow's milk; Hayakawa Y et al.; Exfoliative toxin A (ETA), produced by Staphylococcus aureus, is the causative agent of staphylococcal scalded-skin syndrome (SSSS) in children . Recently, we reported that ETA was detected by reverse passive latex agglutination in three isolates of S . aureus from cow's milk, but that these ETA-positive isolates did not cause the so-called Nikolsky sign in neonatal mice . In this study, therefore, the eta gene encoding ETA and regulatory genes of these bovine isolates were analyzed by the polymerase chain reaction (PCR) and sequencing . The eta gene was amplified from three bovine isolates by PCR and their resulting nucleotide sequences found to correspond to the eta gene from the human isolate, except for three nucleotides in the upstream region of the eta open reading frame (ORF) . An accessory gene regulator (agr), which is a global regulatory locus, was detected in these bovine isolates by PCR amplification . In addition, the ORF (J-4), located 120 bp upstream from the eta ORF of the human isolate, was also amplified from these bovine isolates, with their nucleotide sequences differing at 32 positions from the human isolate . Bovine and human ORF J-4 equally enhanced production of ETA in the recombinants of the eta gene, suggesting that the variation in bovine ORF J-4 may be not be the cause of the difference in amount of ETA produced by bovine and human isolates. Clin Infect Dis, 2001 Jan, 32(1), 108 - 15 Epub 2000 Dec 13. Vancomycin-intermediate and -resistant Staphylococcus aureus: what the infectious disease specialist needs to know; Fridkin SK; Ever since the first strain of Staphylococcus aureus with reduced susceptibility to vancomycin and teicoplanin was reported from Japan, there has been a lot of confusion regarding the laboratory and clinical approach to patients with infections due to S . aureus with reduced susceptibility to vancomycin . To date, 6 clinical infections with vancomycin-intermediate S . aureus (VISA) have been reported in the United States . Intermediate resistance appears to develop from preexisting strains of methicillin-resistant S . aureus in the presence of vancomycin, and all but 1 infection occurred in patients with exposure to dialysis for renal insufficiency . Detection of VISA is difficult in the laboratory, and special inquiries about susceptibility testing methods may be needed . These VISA-infected patients had underlying illnesses, and their infections did not appear to respond well to conventional treatment . Prevention strategies have been outlined . Without continued vigilance in enforcing infection-control measures, improved use of antimicrobials, and coordination of efforts among public health authorities, increasing levels of vancomycin resistance in S . aureus are likely to be encountered. J Nippon Med Sch, 2000 Dec, 67(6), 464 - 7 Septic arthritis of the hip associated with atopic dermatitis . A case report; Kitamura S et al.; We report a case of septic arthritis of the hip associated with atopic dermatitis . A 15-year female felt a pain in the right hip with unknown cause on May 11, 1998 . The pain subsequently became aggravated, and she was admitted to our hospital on May 18 . She has had atopic dermatitis since 4 years of age . She showed generalized dermatitis with desquamation and numerous scratch marks . A culture of both skin and joint fluid revealed Staphylococcus aureus . Physical examination revealed tenderness in Scarpa triangle and restricted range of motion . Immunological serology showed an increase in eosinophils and immunoglobulin E, and a decreased reaction of lymphocyte blastoid transformation . Computed tomography (CT) and MRI showed a joint effusion in the right hip . She was diagnosed as having septic arthritis of the hip . Intravenous drip of Cefazolin of 2g was started on the first day of hospitalization and joint irrigation was done on the second day . CRP became negative at 4 weeks, but joint effusion was shown on CT . Additional joint irrigation with Amicamycin (200 mg) was done . As the joint fluid culture became negative, range of motion exercises were started at 6 weeks . She was discharged with a long-leg brace applied at 8 weeks . At 13 months after onset, she had complete relief of the pain and normal activities of daily living . No destructive changes in the hip were found on X-ray examination or MRI . In the present case, an abnormal immune system associated with atopic dermatitis as well as the habit of scratching eruptions may have led to hematogenous spread of skin infection, and caused septic arthritis of the hip. Chest, 2000 Dec, 118(6), 1832 - 3 Staphylococcus aureus pericarditis masquerading as anterior mediastinal mass: mediastinal mass from pericarditis; Nwiloh JO et al.; Pseudomediastinal mass as a result of bacterial pericarditis is a rare clinical presentation . We report one such case in a patient with end-stage renal disease receiving hemodialysis, who presented primarily with manifestations of right heart compression due to a large encapsulated pericardial abscess and, surprisingly, with no overt signs of sepsis . Surgical drainage, pericardiectomy, and antibiotic therapy led to a successful outcome. Arch Otolaryngol Head Neck Surg, 2000 Dec, 126(12), 1440 - 3 Methicillin-resistant Staphylococcus aureus otorrhea after tympanostomy tube placement: an emerging concern; Hartnick CJ et al.; OBJECTIVES: To review the treatment of pediatric patients with methicillin-resistant Staphylococcus aureus (MRSA)-positive cultures as a result of otorrhea after tympanostomy tube placement in terms of both medication and isolation strategies and to highlight an emerging problem faced by the clinician with reference to treatment options as well as to the treatment of these patients in an outpatient setting . PATIENTS: Between December 1998 and January 2000, a total of 8 children between the ages of 1 and 11 years had MRSA-positive cultures as a result of otorrhea after tympanostomy tube placement . MAIN OUTCOME MEASURES: The Department of Infectious Diseases was notified, and a variety of topical antibiotic treatments were administered . Arch Otolaryngol Head Neck Surg . 2000;126:1440-1443 Mol Microbiol, 2000 Nov, 38(4), 694 - 705 Phage conversion of exfoliative toxin A production in Staphylococcus aureus; Yamaguchi T et al.; The staphylococcal exfoliative toxins (ETs) are extracellular proteins that cause splitting of human skin at the epidermal layer during infection in infants . Two antigenically distinct toxins possessing identical activity have been isolated from Staphylococcus aureus, ETA and ETB . The gene for ETA (eta) is located on the chromosome, whereas that for ETB is located on a large plasmid . The observation that relatively few clinical isolates produce ETA suggests that the eta gene is acquired by horizontal gene transfer . In this study, we isolated a temperate phage (phiETA) that encodes ETA and determined the complete nucleotide sequence of the phiETA genome . phiETA has a head with a hexagonal outline and a non-contractile and flexible tail . The genome of phiETA is a circularly permuted linear double-stranded DNA, and the genome size is 43 081 bp . Sixty-six open reading frames (ORFs) were identified on the phiETA genome, including eta, which was found to be located very close to a putative attachment site (attP) . phiETA converted ETA non-producing strains into ETA producers . Southern blot analysis of chromosomal DNA from clinical isolates suggested that phiETA or related phages are responsible for the acquisition of eta genes in S . aureus. J Bacteriol, 2001 Jan, 183(1), 63 - 70 Characterization of a putative pathogenicity island from bovine Staphylococcus aureus encoding multiple superantigens; Fitzgerald JR et al.; Previous studies have demonstrated that a proportion of Staphylococcus aureus isolates from bovine mastitis coproduce toxic shock syndrome toxin (TSST) and staphylococcal enterotoxin C (SEC) . In this study, molecular genetic analysis of one such strain, RF122, revealed the presence of a 15,891-bp putative pathogenicity island (SaPIbov) encoding the genes for TSST (tst), the SEC bovine variant (sec-bovine), and a gene (sel) which encodes an enterotoxin-like protein . The island contains 21 open reading frames specifying hypothetical proteins longer than 60 amino acids including an integrase-like gene . The element is bordered by 74-bp direct repeats at the left and right junctions, and the integration site lies adjacent to the 3' end of the GMP synthase gene (gmps) in the S . aureus chromosome . SaPIbov contains a central region of sequence identity with the previously characterized tst pathogenicity island SaPI1 (J . A . Lindsay et al., Mol . Microbiol . 29:527-543, 1998) . A closely related strain, RF120, of the same multilocus enzyme electrophoretic type, random amplified polymorphic DNA type, and ribotype, does not contain the island, implying that the element is mobile and that a recent insertion/deletion event has taken place . TSST and TSST/SEC-deficient mutants of S . aureus strain RF122 were constructed by allele replacement . In vitro bovine Vbeta-specific lymphocyte expansion analysis by culture supernatants of wild-type strains and of tst and sec-bovine allele replacement mutants revealed that TSST stimulates BTB13-specific T cells whereas SEC-bovine stimulates BTB93-specific T cells . This suggests that the presence of SaPIbov may contribute to modulation of the bovine immune response. Biochemistry, 2000 Dec 19, 39(50), 15344 - 52 Rhodamine 123 binds to multiple sites in the multidrug resistance protein (MRP1); Daoud R et al.; The mechanisms of MRP1-drug binding and transport are not clear . In this study, we have characterized the interaction between MRP1 and rhodamine 123 (Rh123) using the photoreactive-iodinated analogue, {(125)I}iodoaryl azido-rhodamine 123 (or IAARh123) . Photoaffinity labeling of plasma membranes from HeLa cells transfected with MRP1 cDNA (HeLa-MRP1) with IAARh123 shows the photolabeling of a 190 kDa polypeptide not labeled in HeLa cells transfected with the vector alone . Immunoprecipitation of a 190 kDa photolabeled protein with MRP1-sepcific monoclonal antibodies (QCRL-1, MRPr1, and MRPm6) confirmed the identity of this protein as MRP1 . Analysis of MRP1-IAARh123 interactions showed that photolabeling of membranes from HeLa-MRP1 with increasing concentrations of IAARh123 was saturable, and was inhibited with excess of IAARh123 . Furthermore, the photoaffinity labeling of MRP1 with IAARh123 was greatly reduced in the presence of excess Leukotreine C(4) or MK571, but to a lesser extent with excess doxorubicin, colchicine or chloroquine . Cell growth assays showed 5-fold and 14-fold increase in the IC(50) of HeLa-MRP1 to Rh123 and the Etoposide VP16 relative to HeLa cells, respectively . Analysis of Rh123 fluorescence in HeLa and HeLa-MRP1 cells with or without ATP suggests that cross-resistance to Rh123 is in part due to reduced drug accumulation in the cytosol of HeLa-MRP1 cells . Mild digestion of purified IAARh123-photolabeled MRP1 with trypsin showed two large polypeptides (approximately 111 and approximately 85 kDa) resulting from cleavage in the linker domain (L1) connecting the multiple-spanning domains MSD0 and MSD1 to MSD2 . Exhaustive proteolysis of purified IAARh123-labeled 85 and 111 kDa polypeptides revealed one (6 kDa) and two (approximately 6 plus 4 kDa) photolabeled peptides, respectively . Resolution of total tryptic digest of IAARh123-labeled MRP1 by HPLC showed three radiolabeled peaks consistent with the three Staphylococcus aureus V8 cleaved peptides from the Cleveland maps . Together, the results of this study show direct binding of IAARh123 to three sites that localize to the N- and C-domains of MRP1 . Moreover, IAARh123 provides a sensitive and specific probe to study MRP1-drug interactions. Eur J Drug Metab Pharmacokinet, 2000 Apr-Jun, 25(2), 103 - 8 In vivo metabolism of 4-fluorobenzoic acid {(5-nitro-2-furanyl)methylene} hydrazide in rats; Gulerman NN et al.; It is known that substituted hydrazide hydrazone derivatives have several biological and pharmacological activities; there is limited literature on the metabolism of hydrazide hydrazones in rats . In our previous study, 4-fluorobenzoic acid {(5-nitro-2-furanyl)methylene}hydrazide (S) was found active against Staphylococcus aureus ATCC 29213 . Therefore, we planned to study the in vivo metabolism of S in rats . The substrate was administered in doses of 50 mg/kg or 100 mg/kg intraperitoneally . Blood samples were collected at 0, 5, 15, 30, 45 min and 1, 1.5, 2, 4, 8, 12, 24, 48 h after administration . The substrate and its potential metabolites were separated using HPLC on a reverse phase system . 4-Fluorobenzoic acid and one unidentified metabolite were detected together with substrate. Vet Hum Toxicol, 2000 Dec, 42(6), 341 - 4 Evaluation of the antibacterial and hemolytic activities of Latvian herbal preparation; Atroshi F et al.; Three extracts originating from a combination of various Latvian plant species were tested for their antibacterial activities by evaluating growth delays using a fully automated microturbidimetric method . Ten different human and bovine strains of the genera Staphylococcus and Micrococcus were used as test microorganisms . The inhibitory effect in vitro was defined as the difference between the growth rate without herbs and the growth rate in the presence of an extract . Among the tested strains, Staphylococcus aureus