Staphylococcus aureus

Adv Perit Dial, 1996, 12, 218 - 20
Does the response to hepatitis B vaccination predict CAPD-associated infections?
Holley JL.
Rates of peritoneal dialysis-associated catheter infections and peritonitis were compared in continuous ambulatory peritoneal dialysis patients grouped on the basis of their response to hepatitis B vaccination with Engerix to assess the usefulness of vaccination in predicting patients at risk for peritonitis and catheter infections . Engerix was given intramuscularly in a dose of 40 micrograms at 0, 1, 2, and 6 months . Sixty-three percent (20/32) of patients developed hepatitis B surface antibodies (converters) . Converters and nonconverters were not different in proportions of women, whites, diabetics, or Staphylococcus aureus nasal carriers; mean age and mean months on peritoneal dialysis were also not different . Overall, peritonitis (0.46/year vs 0.33/year) and catheter infection (0.53/year vs 0.54/year) rates were not different among converters and nonconverters, respectively . Nonconverters had higher S . aureus peritonitis rates (0.12/year vs 0.04/year, p < 0.05) but lower S . epidermidis peritonitis rates (0.03/year vs 0.18/year, p < 0.02) . However, when the patient with recurrent S . epidermidis peritonitis was excluded from analysis, S . epidermidis peritonitis rates among converters and nonconverters were not different (0.13/year vs 0.03/year, respectively, p < 0.09) . These data suggest that the development of surface antibodies with hepatitis B vaccination does not predict a reduced risk of S . epidermidis peritonitis . The possibility that nonconverters are more likely to be S . aureus nasal carriers and therefore at greater risk of S . aureus peritonitis deserves further study.

Adv Perit Dial, 1996, 12, 209 - 10
Surgical treatment of persistent exit-site infections; Glickman JD et al.; Persistent exit-site infections and tunnel infections (ESI/TI) are a cause for removal of Swan neck catheters (SNC) . Previous studies report variable success in the treatment of these infections by surgical exposure and removal of the subcutaneous external cuff . We report our experience with this technique . All 5 patients with persistent ESI/TI were successfully treated with antibiotics and surgical intervention . All cultures grew Staphylococcus aureus . Average time to complete healing after surgical exposure was 39.4 days . Mean follow-up after complete healing was 164.8 days . There were no subsequent episodes of ESI/TI in these patients . None of the catheters subsequently malfunctioned or developed leaks . Persistent ESI/TI in Swan neck catheters can be successfully treated with surgical exposure and removal of the subcutaneous external cuff.

J Biomed Mater Res, 1996 Fall, 33(3), 139 - 43
A self-setting TTCP-DCPD apatite cement for release of vancomycin; Hamanishi C et al.; Vancomycin (VCM), a methiciline-cefem resistant Staphylococcus aureus (MRSA)-specific antibiotic, was incorporated in a self-setting tetracalcium phosphate (TTCP)-dicalcium phosphate dihydrate (DCPD) apatite cement that hardened isothermally into a hydroxyapatite (HAP) phase with crystallinity similar to that of host bone . Effective release of VCM into PBS lasted for 2 weeks from cements containing 1% VCM and for longer than 9 weeks from cements containing 5% VCM . The rate of release of VCM differed between cements with different crystallinities as well as between the two dissolution media, PBS and simulated body fluid . Mean concentration of VCM in the bone marrow tissue released from cements containing 5% VCM was 20 times the minimum inhibitory concentration 3 weeks after implantation in bone . Direct contact with new bone was observed with the cements containing 1% VCM . Slow delivery of VCM from a self-setting TTCP-DCPD apatite cement with low crystallinity could be used to treat MRSA osteomyelitis.

Perit Dial Int, 1996, 16 Suppl 3, S51 - S70
Exit-site healing post catheter implantation; Twardowski ZJ et al.; The study goals were (1) to describe the natural healing process post peritoneal dialysis catheter implantation; (2) to discern factors that predispose to exit infection; (3) to recognize signs of early exit-site infection; and (4) to ascertain the influence, if any, of the healing process on subsequent peritonitis rates and final catheter outcomes . There were 226 evaluations of 43 exits {range 3-6 per exit, mean 5.2 + or - 1.1 (SD)} in 41 patients . Eleven exits were in the parasternal area and 32 exits were in the abdomen . Exit sites and sinus tracts were examined weekly for 6 weeks with a magnifying loupe and macro-photographed . Cultures were taken from sterile saline sinus washouts, periexit smears, and nares . Exit sites were categorized into four types: (1) fast-healing exits had no drainage or minimal moisture deep inside by the third week; epidermis started to enter into the sinus within 2-3 weeks, progressed steadily, and covered at least half the visible sinus tract 4-6 weeks after implantation; (2) in slow-healing exits without infection, epidermis started to enter into the sinus after 3 weeks or progressed slowly and did not cover half the visible sinus by 5 weeks; the sinus might have had serous or serosanguineous, but never purulent, drainage persistent up to 4 weeks; (3) healing interrupted by infection initially looked identical to the fast-healing exit, but within 6 weeks the epidermis did not progress or regress, granulation tissue became soft or frankly fleshy; drainage increased and/or became purulent; (4) in slow-healing exits due to early infection, granulation tissue became soft or fleshy and/or drainage became puru lent by 2-3 weeks; sinus epidermization was delayed or progressed slowly, only after infection was appropriately treated . Compared with patients with fast-healing exits, patients with early infected exits were more likely (although not significantly) to be diabetics, to have an abdominal catheter, wound hematoma, higher body mass index, and higher percentage of positive cultures for Staphylococcus aureus in nares . Early colonization of the exit was the most significant factor in determining the healing pattern: the later the colonization, the better the healing . Positive culture from either washout or periexit smear one week after implantation was associated with early exit infection, a higher peritonitis rate, and a high probability of catheter loss due to an exit/tunnel infection, and higher peritonitis rate; however, the time to the first peritonitis episode was not shorter than in the groups with later exit colonization . We postulate that exit infections and peritonitis rates may be decreased by delaying exit colonization using prophylactic antibiotics for at least 2 weeks after implantation and sterile exit dressing procedure for the entire healing time of approximately 6 weeks.

Nippon Jinzo Gakkai Shi, 1996 Jan, 38(1), 27 - 32
{A case of toxic shock syndrome (TSS) induced by methicillin-resistant staphylococcus aureus (MRSA) presenting as acute renal failure with disseminated intravascular coagulation}; Fukuuchi F et al.; A case of a 73-year-old woman with acute renal failure due to toxic shock syndrome (TSS) is reported . The patient was admitted to our hospital with the complaints of high fever, disturbance of consciousness and shock . Laboratory findings on admission were; CRP 25.11 mg/dl, WBC 35000/ microl, Plt 1.6 x 10(4)/ microl, GOT 155 U/l, GPT 65 U/l, CPK 4202 U/l (CPK-MM 96%), BUN 123 mg/dl and SCr 7.0 mg/dl . Because of anuria, hemodialysis was performed . This patient was treated with dopamine, methyl prednisolone (MP), frozen fresh plasma, AT III, antibiotics, and platelet transfusion . The bacterial cultures of blood and cerebrospinal fluid were negative, but MRSA was isolated subsequently from the pharynx and vagina . We investigated the production of toxic shock syndrome toxin 1 (TSST-1) and staphylococcal enterotoxins (SE) . The isolated MRSA produced TSST-1, SEB and SEC . Accordingly, we made the diagnosis of TSS . After improvement of acute renal failure and the patient's general condition, MRSA persisted and TSST-1 was still found in the patient's blood . Finally we eradicated the MRSA and TSST-1 after administration of ciprofloxacin hydrochloride (CPFX) and Rifampicin (RFP).

Biotechnol Prog, 1996 Jan-Feb, 12(1), 77 - 83
Applications of perfluorocarbon affinity emulsions for the rapid isolation of Staphylococcus aureus; McCreath GE et al.; Human immunoglobulin G (IgG) has been immobilized to poly(vinyl alcohol)-stabilized liquid perfluorocarbon droplets . This affinity emulsion has been shown to adsorb Staphylococcus aureus cells from solutions with adsorption capacities of 32 x 10(9) and 48 x 10(9) cells/mL for affinity emulsions with immobilized IgG densities of 1.15 and 2.45 mg/mL, respectively . The kinetics of cell binding from solution were found to be rapid with clearance of S . aureus cells from a suspension (8 x 10(8) cell/mL) achievable in under 5 min . S . aureus cells (1.3 x 10(9) cells) could also be rapidly depleted from a suspension of Saccharomyces cerevisiae (3.4 x 10(10) cell/mL) in under 18 min with no cross-reactivity being observed . The affinity emulsion was stable for at least five cycles of operation with little loss in adsorption capacity when removal of adsorbed cells was carried out at pH 2.5.

Drugs Exp Clin Res, 1996, 22(1), 9 - 15
Potentiation of human polymorphonuclear leukocyte phagocytosis and intracellular bactericidal activity by amoxycillin/clavulanic acid; Cuffini AM et al.; The use of broad spectrum beta-lactamase inhibitors in association with beta-lactam agents provides one strategy to overcome the enzymatic resistance . Clavulanic acid is a potent inhibitor of a wide range of bacterial beta-lactamases and its potentiating effect on amoxycillin has been established both in vitro and in clinical trials . Since the efficacy of an antimicrobial agent in the therapy of infections depends on the interaction of bacteria, antibiotic and phagocytes, we investigated the effect of amoxycillin/clavulanic acid on the in vitro interaction between human polymorphonuclear cells (PMNs) and beta-lactamase producing strains of Klebsiella pneumoniae and Staphylococcus aureus . Clavulanic acid did not have any significant influence upon the PMN phagocytosis and killing against intracellular bacteria . Interestingly, the presence of the suicide inhibitor, with its beta-lactamase inhibitory properties, potentiated the activity of amoxycillin against the beta-lactamase producing strains of K . pneumoniae and S . aureus in such a manner that bacteria became significantly more susceptible to either phagocytosis or microbicidal activities of human phagocytes, compared to both the control and amoxycillin systems.

J Biomater Sci Polym Ed, 1996, 7(10), 905 - 15
Biological activities of new poly(N-1-adamantylmaleimide) and poly(N-1-diamantylmaleimide); Wang JJ et al.; N-1-Diamantylmaleimide was synthesized by reaction of maleic anhydride with 1-aminodiamantane, followed by dehydration with acetic anhydride and sodium acetate . Poly(N-1-adamantylmaleimide) (IIa) and poly(N-1-diamantylmaleimide) (IIb) were synthesized by free radical polymerization . N-1-Adamanlylmaleimide (Ia) and N-1-diamantylmaleimide (Ib), exhibited strong activities in vitro antitumor activities . Interestingly, poly(N-1-adamantylmaleimide) exhibited growth inhibitory values against Colo 205, Hep G2, and SK-BR-3, similar to 5-Fluorouracil . It was noted that poly(N-1-adamantylmaleimide) showed relatively lower cytotoxicity against Molt-4 cells than against Colo 205, Hep G2, and SK-BR-3 cells . The decreasing antitumor activities against individual tumor cell line were in the order Ia > Ib > IIa > IIb . This result shows that N-substituents of maleimides play an important role in their antitumor activity . Additionally, Ia and Ib show good in vitro activity against staphylococcus aureus ATCC 25923 while polymers IIa and IIb exhibited weak activity against S . aureus ATCC 25923.

Eur Spine J, 1996, 5(3), 201 - 3
Spinal epidural abscess: an unusual cause of sciatica; Kotilainen E et al.; A previously healthy patient was admitted to our hospital because of low back pain and sciatica . For 4 weeks preceding the admission, he had been treated with nonsteroidal antiinflammatory analgetics and bed rest with a clinical diagnosis of lumbar disc herniation . On admission, the patient was subfebrile but developed general symptoms of septic infection by the next day . Computed tomography and magnetic resonance imaging of the lumbar spine revealed a spinal epidural abscess and spondylodiscitis at the L5-S1 level . During an emergency laminotomy, gross pus in abundance was evacuated from the epidural space; microbiological cultures from the pus and blood yielded Staphylococcus aureus . The unique clinical presentation of our patient combined with merely indolent symptoms of infection delayed the correct diagnosis . We are not aware of any similar reports of patients with lower spinal epidural abscess whose primary presentation was sciatic pain.

Dermatology, 1996, 192(2), 110 - 5
B-cell chronic lymphocytic leukemia associated with high serum IGE levels and pruriginous skin lesions: successful therapy with IFN-alpha 2b after failure on IFN-gamma; Neuber K et al.; BACKGROUND . Chronic lymphocytic leukemia of the B-cell type (B-CLL) associated with highly elevated serum IgE levels and skin involvement has rarely been observed . Furthermore, not much is known about therapeutic strategies in such diseases . OBJECTIVE . We describe a 56-year-old male patient with a 5-year history of chronic and relapsing pruritic skin lesions as well as recurrent Staphylococcus aureus infections of the skin . RESULTS . Histologically, a lymphocytic and eosinophilic skin infiltrate was seen . Laboratory analysis revealed lymphocytosis (46.5%), eosinophilia (11.9%) as well as markedly elevated serum IgE levels (140,000 IU/ml) . Immunohistology showed dermal B cells with intracytoplasmic IgE . Bone marrow biopsy showed a diffuse infiltrate of small lymphoplasmacytic lymphocytes . Over 80% of blood and bone marrow lymphocytes were CD5 +, CD19 +, CD20 +, CD23 + and CD38 + . Based on these findings, the diagnosis of B-CLL was made . The strong pruritus was resistant to antihistamines and steroids . Therefore, a trial with interferon (IFN) gamma 50 micrograms s.c . daily was started in order to suppress elevated serum IgE but failed . After 1.5 million units s.c . of IFN-alpha 2b every second day for 2 weeks, pruritus and serum IgE levels diminished markedly (48,000 IU/ml) . Skin lesions, pruritus as well as skin infection and serum IgE level improved under continuing IFN-alpha 2b therapy for 2 years . CONCLUSION . In cases with suspected hyper-IgE syndrome, hematological neoplasias have to be excluded . The results argue for a benefical effect of IFN-alpha 2b in suppressing IgE production and symptoms of pruritus in an early-stage B-CLL.

Clin Infect Dis, 1996 Jan, 22(1), 86 - 90
Use of restriction endonuclease analysis of plasmids and pulsed-field gel electrophoresis to investigate outbreaks of methicillin-resistant Staphylococcus aureus infection; Liu PY et al.; We used restriction endonuclease analysis of plasmids (REAP) and pulsed-field gel electrophoresis (PFGE) to investigate clusterings of methicillin-resistant Staphylococcus aureus (MRSA) infections in our orthopedic unit, neurosurgery unit, intensive care unit, and burn unit . Fourteen different strain types were identified by REAP and 10 different strain types were identified by PFGE among 25 MRSA isolates collected during these incidents of infection . Though neither technique was clearly superior to the other for typing MRSA isolates, REAP is recommended as a relatively simple and reproducible technique for the preliminary investigation of MRSA infection outbreaks in clinical settings.

Clin Infect Dis, 1996 Jan, 22(1), 40 - 5
Infective endocarditis in injection drug users: importance of human immunodeficiency virus serostatus and degree of immunosuppression; Pulvirenti JJ et al.; Human immunodeficiency virus (HIV)-infected patients are at increased risk for serious and recurrent bacterial infections . We hypothesized that the degree of immunosuppression may play an important role in outcomes for HIV-seropositive patients with infective endocarditis (IE) . To test our hypothesis, we retrospectively reviewed 144 cases of IE in injection drug users . One hundred two patients with documented HIV status (45 HIV-seropositive patients and 57 HIV-seronegative patients) were included in the analysis . Eleven patients (6 HIV-seropositive patients and 5 HIV-seronegative patients) died in the hospital . Staphylococcus aureus, the most common etiologic pathogen causing IE in our series, was isolated from 32 HIV-seropositive patients (71.1%) and 32 HIV-seronegative patients (56.1%) . A clear inverse correlation between mortality rate and CD4 cell count was demonstrated (r = -.625; P < .001) . Both univariate and multivariate analyses supported the finding of significantly higher mortality rates among patients with CD4 cell counts of < 200/mm3 than among patients with CD4 cell counts of > 500/mm3 (OR, 14.7; 95% CI, 2.64-81.9).

Peptides, 1996, 17(1), 13 - 6
Hydrolysis of N-formylmethionyl chemotactic peptides by endopeptidase 24.11 and endopeptidase 24.15; Lesser M et al.; Endopeptidase 24.11 (EP 24.11), a membrane-bound cell surface enzyme, modulates chemotactic responsiveness of neutrophils to f-Met-Leu-Phe . It is unknown if the enzyme degrades potent formylmethionyl tetrapeptides or if an enzyme with similar activities, endopeptidase 24.15 (EP 24.15), degrades formylated chemotactic peptides . In a study of five formylmethionyl tetrapeptides and f-Met-Leu-Phe, we found that EP 24.11 had high affinity for all peptides evaluated, although it did not effectively degrade f-Met-Ile-Leu-Phe . EP 24.15 had high affinity for three of the tetrapeptides, and for f-Met-Leu-Phe, although, for unclear reasons, it did not degrade f-Met-Ile-Leu-Phe or f-Met-Leu-Phe, the apparent natural products of Staphylococcus aureus and Escherichia coli, respectively.

Zh Mikrobiol Epidemiol Immunobiol, 1996 Jan-Feb, (1), 27 - 30
{The relationship of the indices of solar-geomagnetic activity and the autofluctuations in the biological properties of Staphylococcus aureus 209 subcultures in vitro}; Polikarpov NA; The in vitro study of effect of such factors as solar activity and the magnetic field of the Earth revealed their influence on the autofluctuations of the biological activity of S.aureus 209 subcultures . Differences in the activity of the synthesis of DNAase, RNAase and delatinase could increase two-- to tenfold during the year . Relationships between heliogeophysical characteristics and the biological activity of bacteria could be both direct and reverse . The study also revealed that at the periods of increased solar and geomagnetic activity S.aureus may spontaneously dissociate in the soil, forming subcultures with greater biological activity than the initial strains.

Acta Otolaryngol, 1996 Jan, 116(1), 104 - 11
Interaction and immunological effect of very late antigen-4, 5, and fibronectin in tonsillar lymphocytes and their relation to age; Ohguro S et al.; We examined the expressions of alpha-subunits of very late antigen (VLA)-4 (alpha4) and VLA-5 (alpha5) on tonsillar lymphocytes and the interaction between these integrin receptors and their ligand, fibronectin (FN) . Immunohistological and flow cytometric analyses showed that alpha4 and alpha5 were expressed in the lymphoid follicle and were positive on about 10% each of T cells and on 55% and 35% of B cells . When tonsillar B cells were separated by a discontinuous Percoll gradient, the number of alpha4- and 5-positive cells decreased as the cell density went down, while the number of activated cells went up . After in vitro activation of tonsillar B cells by Staphylococcus aureus Cowan I strain (SAC), the expression of alpha5 and the adhesiveness to FN decreased . The increased proliferation of B cells was observed when tonsillar B cells were cultured with immobilized FN . The expressions of alpha-subunits of VLA-4 and VLA-5 on tonsillar T and B lymphocytes increased with age . These results suggest that: i) B cell activation may cause decreased expressions of VLA-4 and -5, which gives a costimulatory effect on B cell activation itself again in cooperation with FN, ii) Increased expressions of VLA-4 and -5 on tonsillar lymphocytes with age may be related to regional immune response of the palatine tonsils.

Auris Nasus Larynx, 1996, 23, 111 - 20
Expressions of very late antigen-6 and vitronectin receptor, and their interactions to laminin and vitronectin during tonsillar B-cell activation; Ohguro S et al.; This study examined the expressions of a-subunits of very late antigen-6 (VLA-6; alpha 6) and vitronectin receptor (VNR; alpha V) on tonsillar B cells and interactions between those integrins and their respective ligands, laminin (LM) and vitronectin (VN) . alpha 6 and alpha V were expressed on about 30 to 40% of tonsillar B cells . When purified tonsillar B cells were separated by a discontinuous Percoll gradient, the number of alpha 6- and alpha V-positive cells decreased as the cell density went down, while the number of activated cells went up . After in vitro activation of tonsillar B cells by Staphylococcus aureus Cowan I strain (SAC), the expressions of alpha 6 and alpha V and their adhesiveness to LM or VN decreased significantly . Increased proliferation of B cells was observed when tonsillar B cells were cultured with immobilized LM or VN . The results of immunohistological staining showed VLA-6, VNR, LM and VN in the follicular area . These results suggest that the expressions of VLA-6 and VNR on tonsillar B cells may be decreased during B cell activation, and the interaction between VLA-6, VNR, and LM, VN may give a costimulatory effect on B cell activation in the follicular area of the tonsil.

Infect Control Hosp Epidemiol, 1996 Jan, 17(1), 20 - 8
Colonization and infection with methicillin-resistant Staphylococcus aureus: associated factors and eradication; Asensio A et al.; OBJECTIVES: To identify characteristics associated with methicillin-resistant Staphylococcus aureus (MRSA) colonization and infection, and to evaluate the efficacy of systemic and topical antimicrobials in the eradication of MRSA carriage among hospitalized patients . DESIGN: A case-control study was done to identify associations . Odds ratios were estimated by unconditional multiple logistic regression . Cohort study was done to evaluate MRSA decolonization efficacy by an oral regimen . Patients infected or colonized with MRSA received a 5-day course of oral (160 mg/800 mg) trimethoprim-sulfametroxazole twice daily and 600 mg of rifampin once daily as decolonization treatment . The proportion of MRSA-free patients after decolonization treatment was determined . Persistence of clearing was estimated by the Kaplan-Meier method . SETTING: Ramon y Cajal Hospital, a 1,249-bed, tertiary-care teaching hospital in Madrid, Spain . PATIENTS: One hundred ninety-two patients with hospital-acquired MRSA infection/colonization and 195 MRSA-free random controls . RESULTS: Six factors were associated independently with MRSA infection/colonization: age (every 10 years of age, odds ratio {OR} = 1.3); ward (surgical, OR = 1; medical, OR = 3.1; intensive care unit, OR = 60); previous hospitalization (OR = 6.9); coma (OR = 25.3); invasive procedures (each, OR = 1.7); 3 or more weeks of hospitalization (OR = 3.8) . We failed to show antibiotic therapy to be an independent risk factor for MRSA hospital infection/colonization . Overall, MRSA eradication was 64.2% by day 2 to 9 after completion of treatment . Kaplan-Meier product limit survival analysis showed that the probability of remaining MRSA-free was 65.3% (SE = 0.09) at 32 days after completion of treatment . CONCLUSIONS: The results offer a rationale for reduction of MRSA infection/colonization in the hospital by interventions aimed at early identification of patients at higher risk, at prompt discharge of patients, and at preventing dissemination while performing invasive procedures . They also provide estimates of antibiotic treatment efficacy to reduce the reservoir of MRSA in the hospital.

Antimicrob Agents Chemother, 1996 Jan, 40(1), 166 - 72
Overproduction of a 37-kilodalton cytoplasmic protein homologous to NAD+-linked D-lactate dehydrogenase associated with vancomycin resistance in Staphylococcus aureus; Milewski WM et al.; We previously reported the isolation of a laboratory-derived Staphylococcus aureus mutant, 523k, that has constitutive low-level resistance to vancomycin (MIC = 5 micrograms/ml) and teicoplanin (MIC = 5 micrograms/ml) and elaborates a ca . 39-kDa cytoplasmic protein that was not detected in the parent strain 523 (MIC = 1 micrograms/ml) . We have now detected the protein in strain 523 by immunoblotting with antiserum raised against the protein . Consistent with our initial observations, densitometric analysis of the immunoblots revealed an increased production of the protein in 523k compared with that of the susceptible parent 523 . The 5' region of the gene encoding the protein of interest was identified by nucleotide sequencing a PCR product amplified from the genome of 523k with degenerate primers designed to encode the amino acid sequence of proteolytic peptides obtained from the protein . The remainder of the gene was identified by library screening, PCR, and nucleotide sequencing . The gene encodes a 36.7-kDa protein with homology to a family of bacterial NAD+-dependent, D-specific 2-hydroxyacid dehydrogenases which includes both D-lactate dehydrogenase and the enterococcal vancomycin resistance protein VanH and is therefore designated ddh . Increased production of the product of ddh, Ddh, was associated with increased D-lactate dehydrogenase activity in 523k, a finding which suggested that Ddh is likely to be the D-lactate dehydrogenase previously identified in S . aureus . The increased D-lactate dehydrogenase activity in strain 523k and the structural similarities among Ddh, D-lactate dehydrogenase, and VanH suggest that overproduction of Ddh might play a role in vancomycin resistance in this strain.

Neurol Med Chir (Tokyo), 1996 Jan, 36(1), 40 - 4
Cervical discitis associated with spinal epidural abscess caused by methicillin-resistant staphylococcus aureus; Sawada M et al.; A 53-year-old male developed fever, nuchalgia, shoulder pain, and sore throat after a partial sigmoidectomy . He suffered sudden onset of quadriplegia about 5 days later . Magnetic resonance (MR) imaging indicated the characteristic appearance of C5-6 intervertebral disc herniation . However, anterior discectomy showed that he had cervical discitis associated with spinal epidural abscess . Continuous pus drainage from the abscess and vigorous antibiotic therapy were undertaken after surgery . He improved and could walk with assistance . MR imaging with gadolinium can be useful in diagnosing pyogenic spinal infection (epidural abscess, osteomyelitis, and discitis), but care should be taken in MR image interpretation since spinal epidural abscess associated with discitis may mimic disc herniation . Axial MR images are important to distinguish this rare disease from other cervical disorders.

J Ethnopharmacol, 1996 Jan, 50(1), 27 - 34
Comparative study on the antibacterial activity of phytochemical flavanones against methicillin-resistant Staphylococcus aureus; Tsuchiya H et al.; Differently substituted flavanones were isolated from Leguminosae and their antibacterial activity was comparatively studied against methicillin-resistant Staphylococcus aureus (MRSA) . The minimum inhibitory concentrations (MICs) of phytochemical flavanones to clinical isolates of MRSA were determined by a serial agar dilution method . The structure-activity relationship has indicated that 2',4'- or 2',6'-dihydroxylation of the B ring and 5,7-dihydroxylation of the A ring in the flavanone structure are important for significant anti-MRSA activity and that substitution with a certain aliphatic group at the 6- or 8-position also enhances the activity . Among the thirteen flavanones tested, tetrahydroxyflavanones with these structural characteristics isolated from Sophora exigua and Echinosophora koreensis showed intensive activity to inhibit the growth of all MRSA strains at 3.13-6.25 micrograms/ml . The present hydroxyflavanones would be useful in the phytotherapeutic strategy against MRSA infections.

J Biomater Sci Polym Ed, 1996, 7(9), 769 - 80
Bacterial adhesion to polyurethane surfaces in the presence of pre-adsorbed high molecular weight kininogen; Nagel JA et al.; The factors which affect the adherence of a bacteria cell to the surface of a biomaterial include the surface chemistry of the cell and material, as well as the composition of the adsorbed protein layer when the biomaterial is exposed to circulating blood . In an effort to better understand the mechanisms by which bacteria adhere to such surfaces, and specifically to determine the effects of high molecular weight kininogen on bacterial adhesion, experiments were performed in which the attachment of Staphylococcus aureus was directly observed on glass and on a series of functionalized polyurethanes . These surfaces had been pre-adsorbed with various concentrations of high molecular weight kininogen and fibrinogen . Attachment was observed using a radial flow chamber, in which shear stress varied inversely with radial distance . Protein adsorption studies were also performed using 125I labeled fibrinogen to investigate the relationship between surface chemistry, protein adsorption, and bacterial attachment . Bacterial attachment was significantly decreased when the glass surface was pre-adsorbed with high molecular weight kininogen--either alone, or following adsorption of fibrinogen . High molecular weight kininogen thus exhibited anti-adhesive effects . On polyurethane surfaces pre-adsorbed with fibrinogen, kininogen, and albumin, the highest bacterial attachment was found on the base polyurethane, while significant decreases were seen on the hydrophilic polyurethanes . In addition, it was found that the surface with the least bacterial attachment and fibrinogen deposition was the polyurethane with pendant phosphonate groups.

Antibiot Khimioter, 1996 Jan, 41(1), 22 - 6
{Antibioticograms of microorganisms isolated from foci of local infections in infants}; Sentsova TB et al.; Microbiological tests were performed in regard to 474 newborns within 1985-1995 . It was shown that gram-positive microflora (Staphylococcus epidermidis and Staphylococcus aureus) predominated in the etiological structure of omphalitis and conjunctivitis . Among gram-negative isolates in the cases with omphalitis there predominated Klebsiella pneumonia . The antibioticograms were of great practical value for the adequate therapy.

Rev Med Interne, 1996, 17(4), 300 - 4
{Pyomyositis in adults in central Tunisia . Apropos of 10 cases}; Chekir T et al.; Ten adult patients treated for pyomyositis between 1988 and 1994 in Sousse's university hospital (Tunisia) were retrospectively reviewed . Due to the non specific symptoms, the diagnosis was often delayed (mean = 17 days) and other primary diagnoses were considered, mainly including synovitis . The muscles around hip and thigh were most commonly involved (ten patients), and Staphylococcus aureus was the most common pathogen (nine patients) . Ultrasonography was very helpful in the accurate diagnosis of the infection . Incision, drainage, and antibiotic therapy eradicated the infection in all patients . No residual functional limitations and no residual symptoms were noted . Our study showed that pyomyositis is present in central Tunisia and not associated with HIV infection . Clinical features and prognosis are similar to those previously described in the literature.

Chemotherapy, 1996 Jan-Feb, 42(1), 21 - 36
Gentamicin- and methicillin-resistant Staphylococcus aureus: phenotypic and genotypic characterization of three putative nosocomial outbreak strains; Traub WH et al.; Nineteen representative isolates of gentamicin- and methicillin-resistant Staphylococcus aureus (MRSA) were found to comprise three phenotypes; these differed with regard to hydrolysis of nitrocefin and production of staphylococcal enterotoxin A or/and toxic shock syndrome toxin-1 . All MRSA isolates produced a capsule and were susceptible to coumermycin, nitrofurantoin, novobiocin, trimethoprim, trimethoprim-sulfamethoxazole, teicoplanin and vancomycin . All MRSA isolates were resistant to co-amoxiclav, ciprofloxacin, clarithromycin, gentamicin, methicillin, norfloxacin, ofloxacin, oxacillin and polymyxin B.

J Clin Microbiol, 1996 Jan, 34(1), 20 - 4
Clinical comparison of difco ESP, Wampole isolator, and Becton Dickinson Septi-Chek aerobic blood culturing systems; Cockerill FR 3rd et al.; The ESP 80A aerobic blood culture of the ESP automated blood culture system (Difco Laboratories . Detroit, Mich.) was compared with two manual aerobic blood culture systems, the Isolator (Wampole Laboratories, Cranbury, N.J.) and the Septi-Chek (Becton Dickinson, Cockeysville, Md.) systems, for the detection of bloodstream microorganisms from 5,845 blood samples for culture collected from adult patients with suspected septicemia . The bottles were incubated for 7 days, and the sediment from the Isolator tube was inoculated onto solid medium and this medium was incubated for 72 h . A total of 609 microorganisms were recovered from 546 blood cultures . There was no statistically significant difference in the total recovery of microorganisms for the ESP 80A system when compared with that for the Septi-Chek system (P = 0.083); however, the Isolator system recovered significantly more microorganisms overall than either the ESP 80A (P < 0.001) or the Septi-Chek (P < 0.001) system . When assessing individual probable pathogens, the Isolator system detected statistically significantly more Staphylococcus aureus and Candida spp . than either the ESP 80A or the Septi-Chek system (P < 0.05) . Similarly, the Isolator system detected statistically significantly more bloodstream infections (septic episodes) caused by S . aureus and Candida spp . than either the ESP 80A or the Septi-Chek system (P < 0.05) . In blood culture sets which produced growth of the same probable pathogens in the ESP 80A and the Isolator systems, there was no statistically significant difference in the median times to detection for all pathogens combined (P = 0.067) . However, a similar comparison showed the Isolator and the ESP 80A systems to have statistically significantly shorter median detection times for all pathogens combined (P < 0.001) when they were independently compared with the Septi-Chek system . The ESP 80A system had 29 (0.5%) false-positive signals . The ESP system required less processing time than the Isolator system and eliminates the hands-on time for the detection of positive cultures required by the manual systems.

J Laryngol Otol, 1996 Jan, 110(1), 78 - 80
Staphylococcus aureus peritonsillar abscess in an 11-week old infant; Akhtar MJ et al.; A case of Staphylococcus aureus peritonsillar abscess in an 11-week-old infant is described . The importance of peritonsillar abscess culture and its changing management is discussed.

Mem Inst Oswaldo Cruz, 1996 Jan-Feb, 91(1), 101 - 5
Analysis of the clonal diversity of Staphylococcus aureus methicillin-resistant strains isolated at João Pessoa, state of Paraíba . Brazil; Santos Filho L et al.; To investigate the clonal diversity of Staphylococcus aureus strains isolated at Joao Pessoa, State of Paraiba, Brazil, digested genomic DNA were studied by pulsed-field gel electrophoresis (PFGE) in nine methicillin-resistant strains (MRSA) and three methicillin-sensitive strains (MSSA), selected among 67 isolates based on their antimicrobial susceptibility and epidemiology . The isolates were obtained between April and November 1992 from the Hospital of the Federal University of Paraiba, located in Joao Pessoa . Two MRAS isolates from the Oswaldo Cruz Hospital, Sao Paulo, Brazil, including an epidemic strain previously detected from different hospitals at the country were used as control . Five different patterns, were demonstrated by MRSA isolated in Joao Pessoa and these patterns were described in several epidemiologically unrelated hospitals in Sao Paulo . Our results suggest the interstate dissemination of a MRSA clone in Joao Pessoa which is similar to that described in other cities of Brazil.

Perit Dial Int, 1996, 16 Suppl 1, S362 - 7
Impaired initial cell reaction in CAPD-related peritonitis; Koopmans JG et al.; Our objective was to determine the incidence of peritonitis episodes with an impaired initial cell reaction (IICR:neutrophil number < 100 x 10(6)/L) over a period of ten years, and to find possible explanations for this unusual presentation of peritonitis . A retrospective review of the files of continuous ambulatory peritoneal dialysis (CAPD) patients included in the CAPD program 1984 and 1993 was done . Analysis of cytokine and prostanoid patterns during four peritonitis episodes with an IICR was compared to 12 episodes with a normal initial cell reaction (NICR) . Dialysate cell numbers and immunoeffector characteristics of peritoneal cells were compared in 7 IICR patients in a stable situation and a control group of 70 stable CAPD patients . The setting was a CAPD unit in the Academic Medical Center in Amsterdam . Thirty-five CAPD patients who had one or more peritonitis episodes with an IICR and a control group of 249 CAPD patients were included in the study . The incidence of peritonitis with an IICR was 6% . These episodes occurred more than once in 51% of the patients who presented with IICR . In 72% the cell reaction was only delayed: a cell number exceeding 100 x 10(6)/L was reached later . Staphylococcus aureus was significantly more frequently the causative microorganism compared to all peritonitis episodes (PE) that occurred during the study period . Patients with IICR had lower dialysate cell counts in a stable situation, compared to a control group (p < 0.01) . This was caused by a lower number of macrophages and CD4 positive lymphocytes . The phagocytosis capacity of the macrophages appeared to be normal . In a comparison of four PE with an IICR and 12 episodes with an NICR, the tumor necrosis factor-alpha (TNF-alpha) response was similar and occurred on day 1, also pointing to normally functioning macrophages . However, the maximal appearance rates of interleukin-6 (IL-6) and IL-8 occurred later in the episodes with IICR compared to NICR (day 2 vs day 1, p < 0.05) . No differences were found in vasodilating prostaglandins, mesothelial cell markers (cancer antigen 125, phospholipids, hyaluronan), and mesothelial cell numbers in the stable situation nor during peritonitis . Peritonitis can present as abdominal pain in the absence of a cloudy dialysate . In some of the patients this presentation occurred more than once . This impaired, most often delayed, cell reaction was associated with a delayed secondary cytokine response . As IL-6 and IL-8 can be synthesized by mesothelial cells, this suggests an impaired functioning mesothelium . This could not be confirmed, however, by a lower number of mesothelial cells in effluent or lower dialysate levels of mesothelial cell markers.

Enferm Infecc Microbiol Clin, 1996 Jan, 14(1), 27 - 30
{Pyomyositis in Zamora}; Montserrat Chimeno M et al.; BACKGROUND: Pyomyositis is a purulent infection involving the skeletal muscle . Although it was initially described in tropical countries, is it ever more frequently found in warm climates . METHODS: A review of the clinical histories of the admission of patients in the Internal and Infectious Medicine Units over the period from May 1992 to April 1994 (1818 admissions) was carried out analyzing those in whom the diagnosis was of pyomyositis . This diagnosis was performed by clinical data and radiologic and microbiologic confirmation . RESULTS: Five patients diagnosed with pyomyositis were found . Of the five cases, four were men with ages ranging from 27 to 64 years . In most of the cases more than one muscle group was involved and only two cases showed predisposing factors (history of injury and DM) . The existence of abscesses was shown by echography in 3 cases and by CAT in 2 cases . Staphylococcus aureus was the most frequently isolated microorganism . The five patients underwent medical treatment and surgical treatment was also performed in 2 achieving satisfactory evolution . CONCLUSIONS: Imaging techniques (echography and CAT) effectively contribute to the diagnosis of pyomyositis . Medical treatment, associated with surgery in some cases, achieved satisfactory evolution . The incidence of pyomyositis is probably greater than what has been recognized to date.

J Mal Vasc, 1996, 21 Suppl A, 139 - 45
{Antibiotic-impregnated prostheses: eclectic indications}; Melliere D et al.; OBJECTIVE: Infection is a major complication in vascular stents . Stents impregnated with gelatine and dipped in Rifampicin have been shown to resist methicillin-resistant Staphylococcus aureus in both animal experiments and in man . It has been suggested that all aorto-ilio-femoral stents should be treated . To evaluate this method, we reassessed all stent infections observed in our patients who had undergone revascularization of the lower limbs from January 1985 to 1994 . We excluded stents implanted for ruptured aneurysms or implanted in patients with a past history of local infection on vascular stents . RESULTS: The rate of septic complications observed during the first year was 1% for all patients in the series, 0% for aorto-aortic and aorto-biiliac stents and 0.7% for aorto- bifemoral stents . These rates are similar to those reported in the multicentric study directed by Goeau Brissoniere using antibiotic impregnated stents . The extra cost involved in using such stents for aorto-ilio-femoral revascularization was estimated in this series at 2,180,000 Francs . The costs resulting from the three infections was estimated at 960,000 Francs . CONCLUSION: Based on the findings in this series, antibiotic impregnated stents should be indicated only in selected patients due to the extra cost: past history of local infection, ruptured aneurysms, femoro-tibial stents, cross or axillo-femoral revascularization for which the rate of stent infection is 6.3 - 3.2 and 1.4%, immunodeficient patients, multiple reoperations, post-irradiation arteritis and situations known to involve major risk of infection.

Microbiol Immunol, 1996, 40(4), 247 - 54
Isolation and characterization of teichoic acid-lake substance as an adhesin of Staphylococcus aureus to HeLa cells; Matsuura T et al.; A cell wall component that bound to HeLa cells (HeLa cell-binding CWC) was isolated from a clinical isolate of Staphylococcus aureus . The HeLa cell-binding CWC was resistant to heat (100 C, 1 hr) and proteases, did not stain with Coomassie Brilliant Blue R-250 on SDS-PAGE but stained as a broad band with antiserum against the strain on Western blots . These data suggest that the HeLa cell-binding CWC is not a protein, and may be teichoic acid . Purified teichoic acid bound to HeLa cells, whereas fractions without teichoic acid did not . In Western blots, HeLa cell-binding CWC appeared as a broad band of less than 35 kDa, similar to that of purified teichoic acid . These data suggest that the HeLa cell-binding CWC obtained in this study is teichoic acid . Teichoic acid inhibited S . aureus adherence to HeLa cells and bound to the cells time and dose dependently, in a saturable and reversible manner, and therefore appears to be an adhesin of S . aureus to HeLa cells.

Lab Anim, 1996 Jan, 30(1), 28 - 34
Lung, ileum and heart are predilection sites for AApoAII amyloid deposition in CD-1 Swiss mice used for toxicity studies . Pulmonary amyloid indicates AApoAII; Gruys E et al.; Amyloid deposits represent frequent histological findings in SPF strains of mice mainly used for toxicological studies . Usually, these are deposits of reactive amyloid (AA-amyloid) derived from the acute phase protein serum amyloid A (SAA) . The SAA is an apoprotein of high density lipoprotein (apoSAA) . Senescence-accelerated amyloid (ASsam) occurs in a special strain of mice . This type of amyloid is derived from apolipoprotein-AII and, therefore, is called AApoAII . Recently, C57B1/Ka control mice not treated for long duration with immunosuppressive agents, were found to have developed AApoAII-amyloidosis with a predilection for the deposits in the ileum (HogenEsch et al . 1993) . In the present study, SPF CD-1 Swiss outbred mice, used for chronic toxicity experiments, were investigated . Amyloidosis was diagnosed by haematoxylin and eosin staining . The tissue localization of amyloid was recorded and confirmed by Congo red staining . The chemical type of amyloid was investigated by peroxidase antiperoxidase (PAP)-immunostaining using anti-murine AA and anti-murine AApoAII antibodies . Those animals which died during the study and the mice killed at end of the experiment, aged 18 months, from treated as well as non-treated control groups, showed AApoAII-amyloid deposits with similar prevalence . The AApoAII amyloid had organ predilection for gut, heart and lung tissue . A group of animals was euthanazed intercurrently at a young age, since they suffered from spontaneous dermatitis associated with Staphylococcus aureus infection . Sixty-eight percent had reactive amyloid deposits found primarily in spleen, liver, kidney and gut . From these findings and literature data on various other mouse strains, it is concluded that in mice used for toxicity studies, AA and AApoAII types of amyloidosis may be expected . The deposition patterns of these types of amyloid are slightly different . AA-amyloid has a predilection for spleen, liver, gut and kidney, and is often associated with inflammatory lesions of the skin, whereas masses of amyloid in lung, heart and ileum suggest AApoAII . Pulmonary amyloid appears to represent the most reliable deposition criterion for discriminating between both types of amyloidosis.

Vet Microbiol, 1996 Jan, 48(1-2), 51 - 5
Genotyping by PCR, of Staphylococcus aureus strains, isolated from mammary glands of cows; Lipman LJ et al.; A total of 71 Staphylococcus aureus strains isolated from bovine mammary glands were identified and subtyped . The methods used to differentiate between the S . aureus isolates were the DNA polymorphism pattern after amplification with a Polymerase Chain Reaction using several primer combinations and phage typing . The DNA fingerprinting technique using RAPD, ERIC1R and ERIC primers proved to be useful in differentiating isolates of S . aureus . Differentiation of isolates using phage typing gave no additional information compared to the DNA technique . The outbreak of S . aureus in the herd studied was mainly caused by one S . aureus strain . Other strains were only found on three occasions, twice in subclinical infections and once from a case of clinical mastitis . In the latter case the dominant strain was isolated from a different quarter of the same cow . Four of the ten cows studied suffered from clinical mastitis . From those four cows, three remained infected with the same S . aureus strain despite antibiotic treatment.

Arch Med Res, 1996 Summer, 27(2), 165 - 70
Clinical stage, age and treatment in tropical pyomyositis: a retrospective study including forty cases; Martinez-de Jesus FR et al.; A comparative and retrospective trial of 40 patients with tropical pyomyositis covering studies done between January 1, 1987 and November 31, 1990, at the General Hospital at Cosamaloapan, Veracruz, IMSS, was undertaken . The objectives were to compare predisposing factors, clinical data, morbidity, mortaity and hospital stay among 1) medical (group I) and surgical treatment (II), 2) adult and pediatric populations and 3) the clinical stage of the disease (invasive, suppurative and late) . In group I, the family history of diabetes (56%), fever (66%) and hospital stay (6.5 +/- 1.8 days) were significantly different from group II (19%, 100% and 12.8 +/- 5.5 days), respectively . The mean age in adult and pediatric populations was 38 and 8 years, respectively . Pediatric patients had lowest hemoglobin levels (9.7 +/- 1.3) . Upper respiratory antecedent was highest in suppurative stage (65%) . In the late stage eosinophilia (5.9 +/- 6.9), fluctuance muscles (100%), complication rate of 57%, surgical drainage (100%) and mortality of 29% were found . Cultures were performed in 20 cases with negative results in 55% and the remaining 45% were positive to Staphylococcus aureus . Pyomyositis appears to be multifactorial in origin, the antecedents of trauma and upper respiratory infection were the major predisposing factors . Septicemia caused high morbidity and mortality in the late stage . Surgical treatment was frequently needed, increasing costs.

J Dairy Sci, 1996 Jan, 79(1), 62 - 70
Mathematical modeling to estimate efficacy of postmilking teat disinfection in split-udder trials of dairy cows; Lam TJ et al.; A mathematical model was used to estimate the efficacy of postmilking teat disinfection from observations in split-udder trials with natural exposure . Data were studied from an outbreak of Staphylococcus aureus IMI during a split-udder trial in a commercial herd with low SCC . The efficacy of postmilking teat disinfection was similar when calculated based on incidence density rates or on transmission rates of IMI in dipped and control quarters . If, however, first and subsequent S . aureus IMI in a cow were not assumed to be independent and were therefore treated separately in the models, the efficacy of post-milking teat disinfection was calculated as being higher with the modeling procedure . The analysis using mathematical modeling, which includes the effect of the number of existing IMI on the number of new IMI, is presented and discussed . This analysis also allows estimation of the basic reproduction ratio . The impact of postmilking teat disinfection on transmission of pathogens is quantified, and proposals for additional preventive measures can be generated . We concluded that efficacy estimations from split-udder trials, assuming quarters to be independent observations, might underestimate the effect of postmilking teat disinfectants on udder pathogens.

J Int Med Res, 1996 Jan-Feb, 24(1), 12 - 6
Activity of nadifloxacin against methicillin-resistant Staphylococcus aureus isolated from skin infections: comparative study with seven other fluoroquinolones; Nishijima S et al.; The in vitro susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) to nadifloxacin and seven other fluoroquinolones (norfloxacin, ofloxacin, enoxacin, ciprofloxacin, lomefloxacin, tosufloxacin and sparfloxacin) was evaluated . The MRSA isolates were isolated from 114 skin infections between 1991 and 1994 . Nadifloxacin exhibited the lowest minimum inhibitory concentration and there were no MRSA isolates resistant to nadifloxacin, while there were some resistant to all of the other seven fluoroquinolones . The minimum concentrations of these drugs needed to cause 50% inhibition of the isolates increased dramatically from 1991 to 1992, but has hardly changed since 1992.

Hum Immunol, 1996 Jan, 45(1), 42 - 5
The toxic shock syndrome toxin-1 induces anergy in human T cells in vivo; Mahlknecht U et al.; TSST-1 is a Staphylococcus aureus-derived superantigen which has been implicated in the pathogenesis of toxic shock syndrome . In mice, superantigen-induced proliferation is followed by deletion or anergy of reactive T cells . So far, superantigen-induced T-cell anergy has not been observed in humans . We therefore examined PBMCs derived from a 15-year-old patient suffering from severe toxic shock syndrome . Markedly elevated levels of circulating TSST-1-reactive T cells were found by cytofluorometric analysis . Upon in vitro restimulation with TSST-1, hyporesponsiveness of TSST-1-responsive V beta 2+ T cells was detected, thus confirming results obtained in the murine system.

J Antimicrob Chemother, 1996 Jan, 37(1), 65 - 75
Bleomycin resistance in Staphylococcus aureus clinical isolates; Gennimata D et al.; Among clinical isolates of Staphylococcus aureus tested for resistance to the antibiotic bleomycin, 197 were found to be resistant; most of them were also resistant to tobramycin and contained plasmids . DNA dot-blot hybridization analysis of the bleomycin resistant isolates with an 171 bp probe derived from the plasmid pUB110 indicated that 43 strains (22%) carried pUB110-like bleomycin resistance DNA sequences . Analysis of bacterial cell lysates derived from the bleomycin resistant isolates indicated that many contained bleomycin-binding properties (BBP), that prevent DNA damage by the antibiotic . Of 13 strains that were analysed by DNA gel electrophoresis and Southern blot DNA hybridization, six were found to carry pUB110-like bleomycin resistance DNA sequences . These studies indicate that there may be more than one genetic determinant for bleomycin resistance in S . aureus whose DNA is protected.

J Antimicrob Chemother, 1996 Jan, 37(1), 53 - 63
Detection of the mec-A gene and phenotypic detection of resistance in Staphylococcus aureus isolates with borderline or low-level methicillinresistance; Bignardi GE et al.; Eighty-three isolates of Staphylococcus aureus for which MICs of methicillin of 4-16 mg/L had previously been recorded were tested for the presence of the mecA gene with a DNA probe and a PCR assay . There was complete agreement between the results obtained by these methods; 39 isolates were mecA-positive and 44 were mecA-negative . Using the presence of mecA as the defining standard, several phenotypic methods for determining resistance to methicillin were evaluated and a high-inoculum, agar-incorporation breakpoint test was found to offer the best combination of high sensitivity and high specificity . However twenty-seven of the 44 mecA-negative strains were methicillin-resistant according to agar dilution MICs (MIC > 4 mg/L on at least one of the four media used) but none had MICs exceeding 32 mg/L . One of the mecA-positive strains had a methicillin MIC of only 8 mg/L and did not appear to be heteroresistant . The clinical significance of these two groups of 'atypical' isolates may need further investigation . This study highlights the problems of detecting reliably S . aureus with low level methicillin resistance by phenotype methods and the usefulness of direct detection of the mecA gene.

Eur J Clin Microbiol Infect Dis, 1996 Jan, 15(1), 60 - 4
Molecular typing of methicillin-resistant Staphylococcus aureus on the basis of protein A gene polymorphism; Frenay HM et al.; The polymorphic X-region of the protein A gene (spa) was used for molecular typing of methicillin-resistant Staphylococcus aureus (MRSA) strains . The X-region is characterized by a variable number (between 3 and 15) of small repeats . DNA sequencing of MRSA strains revealed 25 distinct repeats . Analysis of MRSA strains grown in vitro and in vivo revealed that the X-region was sufficiently stable for epidemiologic typing of MRSA strains . Spa typing of MRSA strains was compared to phage typing and, in general, concordance was found between the two methods . However, spa typing was more sensitive, allowing differentiation of strains within a particular phage type . Results obtained with spa typing suggest that hospital outbreaks may be caused by two or more MRSA strains . Spa typing may be an important tool in unravelling the spread of MRSA strains within and between hospitals.

J Interferon Cytokine Res, 1996 Jan, 16(1), 7 - 16
Stimulation of natural interferon-alpha/beta-producing cells by Staphylococcus aureus; Svensson H et al.; Human peripheral blood mononuclear cells (PBMCs) produced high levels of antiviral activity, as determined by bioassay, when stimulated by Staphylococcus aureus Cowan I (SAC) and E . coli . Specific immunoassays demonstrated the presence of both IFN-alpha and gamma and, for SAC, also low levels of IFN-beta . The frequencies of SAC-induced IF N-alpha-producing cells (IPCs) were up to 1-2 per 10(3) PBMCs . These IPCs expressed the HLA-DR and CD4 antigens but not CD3, CD14, or CD19, thus resembling the natural IFN-alpha-producing cells (NIPC) . The SAC was more efficient as IFN inducer when heat killed than when streptomycin inhibited . The SAC was inhibitory to virally induced IFN-alpha responses, in particular when streptomycin inhibited . Both pronase treatment and mechanical disruption of SAC cells abolished their capacity to induce IFN-alpha production . Staphylococcal strains lacking or expressing low levels of protein A (SpA) showed a decreased ability to induce IFN-alpha production . However, purified SpA did not itself induce IFN-alpha . Possibly, SpA together with other bacterial surface proteins is important for the capacity of SAC to induce IFN-alpha production in NIPC.

J Formos Med Assoc, 1996 Jan, 95(1), 45 - 50
Home parenteral nutrition in children; Huang FC et al.; From 1985 to 1994, home parenteral nutrition (HPN) was used as the method of feeding nine pediatric patients . Indications for HPN included congenital or acquired short bowel syndrome . Crohn's disease, chronic intractable diarrhea, chronic idiopathic intestinal pseudo-obstruction and Hirschsprung's disease . During the period, two patients died; one of sepsis and the other from hepatic failure . Three of the remaining patients have since moved on to oral feeding, but four patients continued on HPN . The majority of these patients have attained a normal weight and height for age while receiving HPN . All patients were fed via an implanted silicone catheter . Catheters were removed and replaced due to complications including nine episodes of infection, four episodes of occlusion, three episodes of breakage and two episodes of dislodgement . Catheter-related sepsis was the most common cause of morbidity and hospital readmission in patients receiving HPN, but was acceptably rare . Staphylococcus aureus was the most frequently cultured organism from either the catheter tip or the blood . HPN is a relatively safe feeding method for patients who would otherwise remain hospitalized for prolonged periods on parenteral nutrition for permanent or prolonged intestinal failure.

J Clin Periodontol, 1996 Jan, 23(1), 38 - 44
Increased release of free oxygen radicals from peripheral neutrophils in adult periodontitis after Fc delta-receptor stimulation; Gustafsson A et al.; The release of free oxygen radicals and degranulation was studied in neutrophils from 14 patients with adult periodontitis and 14 age- and sex-matched healthy controls . The neutrophils were activated by Fc gamma-receptor stimulation, using Staphylococcus aureus opsonized with gamma globulin . Release of oxygen radicals was measured as luminol-enhanced chemiluminescence . Degranulation was assessed as release of elastase, measured with a specific substrate and as release of lactoferrin measured with ELISA . The neutrophils from the patients showed a significantly higher chemiluminescence and a slightly higher release of elastase, whereas the release of lactoferrin was the same in both groups . In contrast, the ratio between the 2 degranulation products, elastase and lactoferrin, was significantly higher in the group with periodontitis . A flow cytometric analysis of the membrane expression of the adhesion molecules CD 11a, CD 11b, CD 15, CD 16, CD 35 and Mel 14 showed no differences in the median immunofluorescence between the 2 groups . This study showed a more than 2-fold higher release of free oxygen radicals from Fc-gamma-receptor stimulated neutrophils compared with healthy controls, which indicates a specific neutrophil-associated host response in adult periodontitis.

Am J Med, 1996 Jan, 100(1), 24 - 31
Contrasting methicillin-resistant Staphylococcus aureus colonization in Veterans Affairs and community nursing homes; Mulhausen PL et al.; PURPOSE: To compare the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) nares colonization, the patterns of MRSA acquisition, and the risk for subsequent MRSA infection between a hospital-based, Department of Veterans Affairs (VA) nursing home care unit (NHCU) and community-based nursing homes . PATIENTS and METHODS: In this prospective study, 148 residents of three community nursing homes and 55 residents of a VA NHCU had their anterior nares swabbed; repeat cultures were obtained from hospitalized patients and/or individuals colonized with MRSA . Subjects were followed up prospectively for 1 year to note hospitalizations and the development of MRSA infections . RESULTS: The prevalence of MRSA colonization was significantly higher in the VA NHCU than in the community nursing homes (mean +/- SD 30.3% +/- 11% versus 9.9% +/- 4%) . The rate of MRSA nares colonization was similar in the two settings . Acquisition of MRSA took place in both the long-term care facilities and hospitals, with 23.8% of incident cases occurring during a hospitalization . Only 3 of the 27 individuals colonized at baseline developed an MRSA infection . A trend toward an increased rate of infection was seen in colonized individuals residing in the community nursing homes versus those in the VA NHCU (relative risk 4.67; 95% Cl 0.55 to 39.9) . Forty-seven percent of the 55 subjects hospitalized were colonized at some point during the study . In contrast to residents of the VA NHCU, MRSA colonization in the community facilities was a marker for high mortality . CONCLUSIONS: Outcomes from colonization may be different in the VA NHCU population and the community nursing home population.

Ann Emerg Med, 1996 Jan, 27(1), 43 - 8
Contaminated wounds: infection rates with subcutaneous sutures; Mehta PH et al.; STUDY OBJECTIVE: To determine whether buried absorbable subcutaneous sutures (BASS) increase the infection rate in irrigated contaminated wounds . METHODS: This was a randomized, prospective trial in a rat model, with the histologist blinded to treatment group . A single 2-cm dorsal incision was made on each of 30 anesthetized 250-g Sprague-Dawley rats and inoculated with approximately 10(8) organisms of Staphylococcus aureus . After irrigation, 15 wounds were closed with running 4-0 nylon transdermal sutures, and 15 were closed with three interrupted 4-0 coated polyglactin 910 (Vicryl) subcuticular sutures (BASS) and running 4-0 nylon transdermal sutures . On day 7, wounds were scored on a scale of 0 to 3 in six categories: inflammatory infiltrates, fibroplasia and capillary proliferation, necrosis, exudates, giant cells, and edema . The possible range for the cumulative wound score was 0 (no inflammation) to 18 (severe inflammation and infection) . RESULTS: The median total wound score in wounds closed with BASS was 14 (range, 7 to 16); it was 8 (range, 5 to 15) for wounds closed without BASS (P = .0004) . The subscores for inflammation, necrosis, exudate, and edema were also significantly higher in wounds closed with BASS . CONCLUSION: BASS increase the infection rate and the degree of inflammation in contaminated wounds, despite through irrigation.

Scand J Immunol, 1996 Jan, 43(1), 64 - 72
Effect of 12 neutralizing anti-cytokine antibodies on in vitro activation of B-cells . Interleukin-12 is required by B1a but not B2 cells; Jones BM; Normal human peripheral blood mononuclear cells, depleted of most monocytes and virtually all CD8-positive cells, were stimulated in vitro with pokeweed mitogen plus Staphylococcus aureus Cowan I in the presence or absence of various neutralizing anti-cytokine antibodies . Numbers of CD5+ and CD5- immunoglobulin-secreting cells were determined using the protein A haemolytic plaque assay after labelling B1a cells with anti-CD5-coated beads . Antibodies against IL-2, IL-5 and IL-10 had little or no effect on plaque-forming cell (PFC) induction; anti-IL-6, -TNF alpha and -TGF beta enhanced PFC induction; anti-IL-1 alpha, -IL-1 beta, -IL-4, -IFN gamma and -IL-13 suppressed PFC induction . B1a and B2 cells were equally affected by cytokine deprivation using these 11 neutralizing antibodies . In contrast, neutralizing anti-IL-12 suppressed induction of CD5+ but not CD5- PFC . Furthermore, recombinant IL-12, if added during the first 48 h of culture, enhanced CD5+ PFC induction while marginally suppressing (IgG-) or not affecting (IgA-, IgM-) induction of CD5- PFC . IL-12 did not preferentially increase survival in culture of B1a cells nor induce expression of CD5 on B2-cells . Further studies are required to determine whether manipulation of B1a and B2 subsets in vivo using IL-12 could be achieved in clinical situations where imbalances in the two populations have been observed.

Scand J Immunol, 1996 Jan, 43(1), 23 - 30
The influence of follicular dendritic cells on B-cell proliferation depends on the activation of B cells and the mitogen used; Bosseloir A et al.; Follicular dendritic cells (FDC) are unique non-lymphoid cells found only in lymph follicles . They play a part in the survival, proliferation and differentiation of B cells . To analyse the influence of FDC on B-lymphocyte proliferation, we isolated them from human tonsils on albumin gradients and treated them with mitomycin C to prevent the multiplication of lymphoid cells harboured in their cytoplasmic evaginations . FDC cultured for 12-16 h remained attached to the substrate; non-adherent cells were carefully eliminated by washing . Purified B cells cultured alone or with contaminant-cleared FDC were maintained for 2 days in the presence or absence of various stimulants, after which tritiated thymidine uptake by these cells was measured . In the absence of activators, FDC did not induce B-cell multiplication . B cells cultured in the presence of FDC exhibited increased 3H-TdR uptake when activated with anti-CD40 MoAb, anti-immunoglobulin MoAb or transferrin, but not when stimulated with Staphylococcus aureus strain Cowan I (SAC) at a given concentration . In the latter case, B-cell proliferation clearly decreased . In control cocultures where mitomycin-C-treated non-adherent cells were used instead of FDC in the presence of the different stimulants, no increase in B-cell proliferation was observed . The results suggest that, inside the germinal centres, FDC modulation of B-cell proliferation depends on the activation state of the B cells and on the stimulant encountered.

Infect Immun, 1996 Jan, 64(1), 310 - 8
Staphylococcus aureus binding to human nasal mucin; Shuter J et al.; Colonization of human nasal mucosa with Staphylococcus aureus sets the stage for subsequent systemic infection . This study characterizes S . aureus adhesion to nasal mucosa in vitro and investigates the interaction of S . aureus with human nasal mucin . S . aureus binding to cell-associated and cell-free mucus was greater than to nonmucin-coated epithelial cells . Scanning electron microscopy of S . aureus incubated with human nasal mucosal tissue showed minimal binding to ciliated respiratory epithelium . In a solid-phase assay, S . aureus bound to purified human nasal mucin-coated wells significantly more than to bovine serum albumin-coated microtiter wells . Binding to mucin was saturable in a dose- and time-dependent fashion . Staphylococcal adherence to human nasal mucin was inhibited by bovine submaxillary mucin but not by fibrinogen . Pretreatment of mucin with periodate but not with pronase reduced adherence . Trypsin treatment of the bacteria significantly reduced adherence to mucin . 125I-labelled nasal mucin bound to two surface proteins (138 and 127 kDa) of lysostaphin-solubilized S . aureus . Binding to human nasal mucin occurs in part via specific adhesin-receptor interactions involving bacterial proteins and the carbohydrate moiety in mucin . These experiments suggest that S . aureus binding to mucin may be critical for colonization of the nasopharyngeal mucosa.

J Clin Invest, 1996 Jan 1, 97(1), 250 - 7
The potent anti-Staphylococcus aureus activity of a sterile rabbit inflammatory fluid is due to a 14-kD phospholipase A2; Weinrauch Y et al.; The cell-free fluid (ascitic fluid, AF) of a sterile inflammatory peritoneal exudate elicited in rabbits is potently bactericidal for complement-resistant gram-negative as well as gram-positive bacterial species . This activity is absent in plasma . We now show that essentially all activity in AF against Staphylococcus aureus is attributable to a group II 14-kD phospholipase A2 (PLA2), previously purified from AF in this laboratory . Antistaphylococcal activity of purified PLA2 and of whole AF containing a corresponding amount of PLA2 was comparable and blocked by anti-AF-PLA2 serum . At concentrations present in AF (approximately 10 nM), AF PLA2 kills > 2 logs of 10(6) S . aureus/ml, including methicillin-resistant clinical isolates, and other species of gram-positive bacteria . Human group II PLA2 displays similar bactericidal activity toward S . aureus (LD90 approximately 1-5 nM), whereas 14-kD PLA2 from pig pancreas and snake venom are inactive even at micromolar doses . Bacterial killing by PLA2 requires Ca2+ and catalytic activity and is accompanied by bacterial phospholipolysis and disruption of the bacterial cell membrane and cell wall . These findings reveal that group II extracellular PLA2, the function of which at inflammatory sites has been unclear, is an extraordinarily potent endogenous antibiotic against S . aureus and other gram-positive bacteria.

J Bacteriol, 1996 Jan, 178(2), 441 - 6
Cell wall sorting of lipoproteins in Staphylococcus aureus; Navarre WW et al.; Many surface proteins are thought to be anchored to the cell wall of gram-positive organisms via their C termini, while the N-terminal domains of these molecules are displayed on the bacterial surface . Cell wall anchoring of surface proteins in Staphylococcus aureus requires both an N-terminal leader peptide and a C-terminal cell wall sorting signal . By fusing the cell wall sorting of protein A to the C terminus of staphylococcal beta-lactamase, we demonstrate here that lipoproteins can also be anchored to the cell wall of S . aureus . The topology of cell wall-anchored beta-lactamase is reminiscent of that described for Braun's murein lipoprotein in that the N terminus of the polypeptide chain is membrane anchored whereas the C-terminal end is tethered to the bacterial cell wall.

J Bacteriol, 1996 Jan, 178(2), 418 - 23
Characterization of the sar locus and its interaction with agr in Staphylococcus aureus; Heinrichs JH et al.; The expression of cell wall and extracellular proteins in Staphylococcus aureus is controlled by global regulatory systems, including sar and agr . We have previously shown that a transposon insertion into the 372-bp sarA gene within the sar locus resulted in decreased expression of several extracellular and cell wall proteins (A . L . Cheung and S . J . Projan, J . Bacteriol . 176:4168-4172, 1994) . In this study, Northern (RNA blot) analysis with a 732-bp sarA probe indicated that two major transcripts (0.56 and 1.2 kb) were absent in the sar mutant compared with the parental strain RN6390 . Additional transcriptional studies revealed that the sarA gene is encoded within the 0.56-kg transcript . Notably, a plasmid carrying the sarA gene together with a 1.2-kb upstream fragment (1.7 kb total) was able to reestablish the 1.2-kb transcript in the mutant . Although reconstitution of the parental phenotype by the sarA gene was incomplete, the introduction of a plasmid carrying the 1.7-kb fragment to the mutant restored the parental phenotype . Transcription of RNAII and RNAIII, which encode the structural and regulatory genes of agr, respectively, was diminished in the mutant but restored to wild-type levels by complementation with the 1.7-kb fragment . In gel shift assays, cell extracts of this clone were able to retard the mobility of a labeled RNAII promoter probe but not an RNAIII promoter element . These data suggest that sarA and the adjacent upstream DNA are essential to the expression of a DNA-binding protein(s) with specificity for the RNAII promoter, thereby controlling agr-related transcription.

Arch Dermatol, 1996 Jan, 132(1), 27 - 33
Staphylococcal enterotoxin B applied on intact normal and intact atopic skin induces dermatitis; Strange P et al.; BACKGROUND AND DESIGN: Colonization of inflammatory skin diseases with Staphylococcus aureus is a frequent phenomenon and may cause exacerbation of the skin disease . Staphylococcus aureus strains present on atopic dermatitis are capable of releasing staphylococcal enterotoxins, a group of superantigens that are very potent T-cell activators . To determine whether the superantigen staphylococcal enterotoxin B can induce inflammation when applied on the skin, staphylococcal enterotoxin B was applied with and without occlusion on the volar aspect of the skin on the forearm of 10 subjects without skin disease and six subjects with atopic dermatitis of minimal activity and no eczema on the volar aspect of the skin on their forearm . The main outcome measures were clinical rating; determination of the increase of the thickness of the skin-fold; and determination of skin blood flow . RESULTS: Clinically, staphylococcal enterotoxin B induced skin changes of erythema and induration in 10 of 10 healthy volunteer subjects and six of six subjects suffering from atopic dermatitis, while the vehicle induced clinically evident skin changes in only one of 10 healthy subjects and none of six subjects with atopic dermatitis . On day 3 after the application of an occluded patch containing 10 micrograms/cm2 of staphylococcal enterotoxin B in the healthy subjects, the thickness of the skinfold increased 0.47 +/- 0.49 mm (mean +/- SD) (n = 9; P < .02) relative to the increase in the thickness of the skinfold following application of the vehicle . The Doppler laser-measured skin blood flow index had increased from 1.0 +/- 0.4 to 5.3 +/- 3.7 (mean +/- SD) (n = 10; P < .002) . On day 3 after the application of occluded patchs containing 10 micrograms/cm2 of staphylococcal enterotoxin B in the subjects suffering from atopic dermatitis, the increase in the thickness of the skinfold increased 0.20 +/- 0.24 mm (n = 6; P, not significant) relative to the increased thickness in the skinfold following application of the vehicle . The Doppler laser-measured skin blood flow index had increased from 1.1 +/- 0.4 to 3.7 +/- 2.2 (n = 6, P, not significant) . Three of six subjects suffering from atopic dermatitis experienced a flare of their disease in the elbow flexure ipsilaterally to where the staphylococcal enterotoxin B patch was applied . CONCLUSIONS: The superantigen staphylococcal enterotoxin B applied on intact skin from both normal subjects and patients with atopic dermatitis induces an inflammatory reaction . This finding suggests that superantigens released from S aureus present on the skin in inflammatory skin diseases may exacerbate and sustain the inflammation.

Am J Kidney Dis, 1996 Jan, 27(1), 75 - 83
Spinal epidural abscess in hemodialysis patients: report of three cases and review of the literature; Obrador GT et al.; Spinal epidural abscess (SEA) is a rare infection that can evolve to severe permanent neurologic deficit or death if the diagnosis is delayed . We report three cases of SEA in hemodialysis patients and summarize nine previously reported cases occurring in hemodialysis patients, with detailed comparisons to series of SEA from the general medical literature . Among these 12 patients, hemodialysis catheters and arteriovenous grafts were the major source of infection, in contrast to the usual skin source . Staphylococcus aureus was implicated in all cases, but in one patient Bacteroides fragilis also was isolated from both the resected arteriovenous fistula and the SEA, and Escherichia coli was isolated from the arteriovenous fistula . The classic syndrome of SEA includes fever, backache, and local spinal tenderness, followed by progressive radicular and cord compression signs and symptoms . In this series, back pain and radicular pain were common at presentation, but only a minority had fever, back tenderness, weakness, or leukocytosis . Cerebrospinal fluid was typically abnormal but culture negative, whereas blood and epidural abscess cultures were frequently positive . Plain x-ray films, bone scans, and plain computed tomography scans had low diagnostic yield, and magnetic resonance imaging with gadolinium had a sensitivity of 80% . Only myelography or computed tomography-myelography gave consistently correct diagnoses . The clinical outcome was poor, with one patient deceased and seven with severe weakness or paralysis . Early intervention provided a higher likelihood of good outcome, whereas late intervention and preoperative neurologic deficits portended a poor functional result . Because of the high incidence of bacteremia in hemodialysis patients, we recommend that symptoms of fever, backache, and spinal tenderness be promptly evaluated for SEA before signs or symptoms of cord compression develop . Early recognition and treatment with antibiotics and decompressive laminectomy is generally associated with a better outcome.

J Infect Dis, 1996 Jan, 173(1), 98 - 103
Effect of tampon composition on production of toxic shock syndrome toxin-1 by Staphylococcus aureus in vitro; Parsonnet J et al.; To determine whether tampons composed of cotton or rayon differ in their effects on production of TSST-1 in vitro, two methods of bacterial cultivation, with and without agitation, were used to study the effects of 16 commercially available and experimental tampons on production of TSST-1 . Experiments were done in blinded fashion . TSST-1 was produced in medium alone in the absence of a tampon by both methods . Neither cotton nor rayon tampons consistently increased toxin production, nor were there significant differences between them in their effects on TSST-1 . In contrast, a tampon composed of carboxymethylcellulose and polyester foam increased production of TSST-1 to a large degree in both culture systems . Thus, neither cotton nor rayon amplifies production of TSST-1 in vitro, and cotton tampons cannot be claimed to be inherently safer on the basis of such data.

J Infect Dis, 1996 Jan, 173(1), 203 - 11
Treatment of experimental gram-negative and gram-positive bacterial sepsis with the hematoregulatory peptide SK&F 107647; DeMarsh PL et al.; SK&F 107647, a novel synthetic low-molecular-weight peptide, has demonstrated potent antiinfective activities in murine models of fungal and viral infection . To determine if the hematoregulatory activities of SK&F 107647 could offer protection over conventional antibiotic therapy or as a single agent in animal models of bacterial sepsis, rats were implanted intraperitoneally with a live bacteria-containing fibrin-thrombin clot . Rats pretreated subcutaneously or orally with SK&F 107647 and then infected with either a gram-negative (Escherichia coli) or a gram-positive (Staphylococcus aureus) bacteria-containing clot demonstrated significantly improved survival over control formulation-treated animals . Treated animals showed increased effector cell activation, measured by CD11b expression on neutrophils and monocytes, and up to 1000-fold reduction in the number of E . coli recovered from blood . Thus, the hematoregulatory activities of SK&F 107647 can increase natural host resistance to infections caused by both gram-negative and gram-positive bacteria.

Gene, 1995 Dec 29, 167(1-2), 115 - 9
Association of staphylococcal type-1 capsule-encoding genes with a discrete genetic element; Lee CY; Previous studies have shown that 13 genes located in a 14.6-kb region of the chromosome of Staphylococcus aureus (Sa) M are required for type-1 capsular polysaccharide (CP1) biosynthesis . In this report, a total of 17 Sa strains producing different CP serotypes were analyzed by Southern hybridization using DNA probes from the cap1 coding region and the flanking sequences . The results showed that the sequence encoding cap1 genes was specific to CP1-producing strains . In addition, DNA regions of at least 18 kb flanking the cap1 genes were absent in most of the non-type-1 strains . These data suggest that the cap1 genes are associated with a chromosomally located discrete genetic element . One end of the element, referred to as the cap1 element, is located in a 1.7-kb fragment about 11.1 kb upstream from the first gene of the cap1 locus and the other end is located in a 0.8-kb region about 7.6 kb downstream from the last gene of the cap1 locus . Thus, the size of the cap1 element is between 33.3 and 35.8 kb.

Gene, 1995 Dec 29, 167(1-2), 111 - 3
Analysis of Staphylococcus aureus genes encoding penicillin-binding protein 4 and an ABC-type transporter; Domanski TL et al.; A region of the Staphylococcus aureus chromosome has been isolated that contains the gene encoding penicillin-binding protein 4 (PBP4), as well as abcA, a gene that encodes a protein with strong sequence similarity to the ABC transporter family of proteins . A disruption in abcA by Campbell-type integration results in cells that display an increased resistance to methicillin and cefoxitin, two antibiotics known to interact with low-molecular-weight PBPs . Based on these observations, a potential regulatory link between these two genes is discussed.

Proc Natl Acad Sci U S A, 1995 Dec 19, 92(26), 12055 - 9
Cell density control of staphylococcal virulence mediated by an octapeptide pheromone; Ji G et al.; Some bacterial pathogens elaborate and secrete virulence factors in response to environmental signals, others in response to a specific host product, and still others in response to no discernible cue . In this study, we have demonstrated that the synthesis of Staphylococcus aureus virulence factors is controlled by a density-sensing system that utilizes an octapeptide produced by the organism itself . The octapeptide activates expression of the agr locus, a global regulator of the virulence response . This response involves the reciprocal regulation of genes encoding surface proteins and those encoding secreted virulence factors . As cells enter the postexponential phase, surface protein genes are repressed by agr and secretory protein genes are subsequently activated . The intracellular agr effector is a regulatory RNA, RNAIII, whose transcription is activated by an agr-encoded signal transduction system for which the octapeptide is the ligand.

FEMS Microbiol Lett, 1995 Dec 15, 134(2-3), 209 - 12
Triton X-100 alters the resistance level of methicillin-resistant Staphylococcus aureus to oxacillin; Komatsuzawa H et al.; We used population analysis to examine the effects of Triton X-100 on the level of resistance to oxacillin of 18 methicillin-resistant Staphylococcus aureus . In the presence of 0.02% Triton-X-100, 17 formerly methicillin-resistant strains exhibited enhanced sensitivity to oxacillin . One homogenous isolate, KSAF1 was barely affected by the Triton X-100 . Sensitivities of lysostaphin, 51 kDa N-acetylglucosaminidase and 62 kDa N-acetylmuramoyl-L- alanine amidase to heat-inactivated cells were not affected when the bacteria were grown in 0.02% Triton X-100 . Our data, together with those of a previous study, suggested that Triton X-100 alters the resistance level of methicillin-resistant S . aureus influencing a factor (s) other than PBPs, bacteriolytic enzymes, or femAB products.

J Biol Chem, 1995 Dec 15, 270(50), 29923 - 7
Moricin, a novel type of antibacterial peptide isolated from the silkworm, Bombyx mori; Hara S et al.; A novel antibacterial peptide that shows antibacterial activity against Staphylococcus aureus was isolated from the hemolymph of the silkworm, Bombyx mori . The novel peptide consisted of 42 amino acids and was highly basic . This peptide indicated no significant similarity with other antibacterial peptides . The peptide showed antibacterial activity against several Gram-negative and -positive bacteria and had a higher activity against Gram-positive bacteria than cecropin B1, a major antibacterial peptide of B . mori . The novel peptide was inducible by bacterial injection . These results suggest that the peptide is responsible for the antibacterial activity in B . mori against Gram-positive bacteria . The effects of the peptide on bacterial and liposomal membranes showed that a target of the peptide is the bacterial cytoplasmic membrane . The results also suggest that the N-terminal portion of the peptide, containing a predicted alpha-helix, is responsible for an increase in the membrane permeability . We propose the name "moricin" for this novel antibacterial peptide isolated from B . mori.

J Biol Chem, 1995 Dec 15, 270(50), 29854 - 61
Molecular cloning, expression, and partial characterization of two novel members of the ovalbumin family of serine proteinase inhibitors; Sprecher CA et al.; A human placental lambda gt11 cDNA library was screened for sequences encoding proteins related to human proteinase inhibitor 6 (PI6), and two plaques were identified that displayed weak hybridization at high stringency . Isolation and characterization of the DNA inserts revealed two novel sequences encoding proteins composed of 376 and 374 amino acids with predicted molecular masses of approximately 42 kDa . The novel proteins displayed all of the structural features unique to the ovalbumin family of intracellular serpins including the apparent absence of a cleavable N-terminal signal sequence . The degree of amino acid sequence identity between the novel serpins and PI6 (63-68%) significantly exceeds that of any other combination of known intracellular serpins . The two novel serpins encoded by the two novel cDNA sequences have been designated as proteinase inhibitor 8 (PI8) and proteinase inhibitor 9 (PI9) . The putative reactive center P1-P1' residues for PI8 and PI9 were identified as Arg339-Cys340 and Glu340-Cys341, respectively . PI9 appears to be unique in that it is the first human serpin identified with an acidic residue in the reactive center P1 position . In addition, the reactive center loop of PI9 exhibits 54% identity with residues found in the reactive center loop of the cowpox virus CrmA serpin . Two PI8 transcripts of 1.4 kilobases (kb) and 3.8 kb were detected by Northern analysis in equal and greatest abundance in liver and lung, while the 1.4-kb mRNA was in excess over the 3.8-kb mRNA in skeletal muscle and heart . Two PI9 transcripts of 3.4 and 4.4 kb were detected in equal and greatest abundance in lung and placenta and were weakly detected in all other tissues . PI8 and PI9 were expressed in baby hamster kidney and yeast cells, respectively . Immunoblot analyses using rabbit anti-PI6 IgG indicated the presence of PI8 in the cytosolic fraction of stably transfected cells that formed an SDS-stable 67-kDa complex with human thrombin . PI9 was purified to homogeneity from the yeast cell lysate by a combination of heparin-agarose chromatography and Mono Q fast protein liquid chromatography and migrated as a single band in SDS-polyacrylamide gel electrophoresis with an apparent molecular mass of 42 kDa . Purified recombinant PI9 failed to inhibit the amidolytic activities of trypsin, papain, thrombin, or Staphylococcus aureus endoproteinase Glu-C and did not form an SDS-stable complex when incubated with thrombin . The cognate intracellular proteinases that interact with PI8 and PI9 are unknown.

J Immunol, 1995 Dec 15, 155(12), 5795 - 802
Collagen-like complement component C1q is a membrane protein of human monocyte-derived macrophages that mediates endocytosis; Kaul M et al.; The collagen-like C1q molecule, a subcomponent of the first component of complement, C1, is synthesized by macrophages (M phi) . Previously, we have demonstrated that C1q is a membrane protein of guinea pig peritoneal macrophages (M phi) . To extend this observation as a general biologic characteristic of M phi, we investigated human (hu) monocyte-derived M phi . Interestingly, surface labeling with the biotin derivative sulfosuccinimidyl-6-(biotinamido)-hexanoate of M phi, freshly isolated monocytes, lymphocytes, granulocytes, and myelomonocytic U937 cells revealed that C1q occurs only on the surface of M phi and not on the surface of the other cells types . Therefore, C1q appears to be a marker for differentiation into M phi . FITC-labeled, fixed Staphylococcus aureus coupled to membrane C1q via a monoclonal alpha-hu-C1q Ab were used to demonstrate that membrane C1q is capable of mediating phagocytosis . Various detergents (Nonidet P-40, digitonin, lubrol, and Triton X-114) were used to solubilize membrane C1q . Membrane C1q of hu M phi is tightly bound to or located in the intact membrane, since treatment of cells with acidic buffers ("acid strip") failed to remove C1q from the cell surface . However, repeated freezing and thawing of cells and washing of segregated membranes with buffer containing 1 M KCl and 3 M urea brought about a marked release of membrane C1q.

Br J Nurs, 1995 Dec 14-1996 Jan 11, 4(22), 1345 - 9
Methicillin-resistant Staphylococcus aureus and wound management; Phillips E et al.; The incidence of methicillin-resistant Staphylococcus aureus (MRSA) infection is increasing in the UK . MRSA is mainly transmitted via the hands of health-care workers . MRSA can cause colonization or infections . MRSA infections can be fatal.

FEBS Lett, 1995 Dec 4, 376(3), 135 - 40
Characterisation of a synergohymenotropic toxin produced by Staphylococcus intermedius; Prevost G et al.; Staphylococcal synergohymenotropic (SHT) toxins damage membranes of host defence cells and erythrocytes by the synergy of two secreted and non-associated proteins: class S and class F components . Whereas Panton-Valentine leucocidin (PVL), gamma-hemolysin and Luk-M from Staphylococcus aureus are members of this toxin family, a new bi-component toxin (LukS-I + LukF-I) from Staphylococcus intermedius, a pathogen for small animals, was characterised and sequenced . It is encoded as a luk-I operon by two cotranscribed genes, like PVL, LukS-I + LukF-I shares a strong leukotoxicity of various PMNs, but only slight haemolytic properties on rabbit erythrocytes . When intradermally injected into rabbit skin, a 100 ng dose caused acute inflammatory reaction leading to tissue necrosis . The new SHT seemed to be largely distributed among various Staphylococcus intermedius strains.

J Gastroenterol, 1995 Dec, 30(6), 718 - 24
The order of ward rounds influences nosocomial infection . A 2-year study in gastroenterologic surgery patients; Yoshida J et al.; We studied the effect of the order of ward rounds on nosocomial infection in gastroenterologic surgery patients . The subjects were patients with gastrointestinal diseases admitted between September 1992 and August 1994 . During the 1st year, the round proceeded indiscriminately among recovery rooms and rooms with stable patients and isolated patients with methicillin-resistant Staphylococcus aureus (MRSA) . In the subsequent year, the round started in the recovery rooms and moved into the general rooms with stable patients and finally into the isolation rooms . Against the time course, piecewise linear regression analyses were made with the number of culture-positive patients and the quantities of antibiotics and disinfectants used . Of a total of 1894 strains from 264 patients, isolates of MRSA (n = 200) decreased from 150 in the 1st year to 50 in the 2nd year . The number of MRSA-positive patients showed the point of inflexion in the analysis at the change of round order, with a later decrease . The trend was similar for Candida (n = 99) and Enterococcal (n = 225) species . The amount of antibiotics was unchanged while the amount of disinfectants used decreased in the 2nd year . Thus, the round re-ordering appeared to help prevent nosocomial infection . Ward rounds for patients who have had gastroenterologic surgery should proceed from compromised hosts to stable patients, and then isolated patients.

J Hosp Infect, 1995 Dec, 31(4), 253 - 60
Molecular epidemiology of a multiple strain outbreak of methicillin-resistant Staphylococcus aureus amongst patients and staff; Lessing MP et al.; Since methicillin-resistant Staphylococcus aureus (MRSA) isolates are not endemic in our hospital, which is a tertiary referral centre, the finding of 13 MRSA isolates from 12 patients associated with an acute vascular surgery ward between October 1993 and December 1993 prompted further epidemiological and laboratory investigations . Two strains were distinguished by antibiogram and phage-typing . One strain, resembling EMRSA-16, colonized six patients and was probably introduced from another hospital in the Oxford Region . Five other patients were colonized by a second strain, gentamicin-resistant and non-typable by phage-typing, probably introduced into the hospital 12 months previously by a patient from Nairobi, Kenya . A 12th patient was colonized by both strains simultaneously . Of 46 staff members screened three were colonized--one by an EMRSA-16 strain, a second by the gentamicin-resistant 'Nairobi'-strain and a third member carried yet a further distinct MRSA strain . The healthcare worker colonized by the 'Nairobi'-strain had been carrying the isolate 12 months previously and was the likely source of this strain . These isolates were also characterized by the repetitive extragenic palindromic-polymerase chain reaction (REP-PCR), a novel PCR-based methodology which has not been previously used in characterizing Staphylococcus aureus in an outbreak . This method corroborated the strain classifications provided by the traditional methods, confirming that there had been spread of two strains simultaneously . Our study demonstrates that multiple strains of MRSA may circulate amongst patients and staff during an outbreak, patients may be colonized by more than one strain simultaneously and long-term staff carriage (> 12 months) may be an important source of colonization in patients . REP-PCR is a rapid and effective molecular typing method for MRSA.

Rev Esp Fisiol, 1995 Dec, 51(4), 213 - 8
Serum hemolytic and bactericidal activity in breast and formula-fed infants; Barriga C et al.; Whether formula or breast feeding influences the functional activity of the complement system from birth to three months of age has been studied . The classical pathway was evaluated by assessing hemolytic activity, based on the capacity of the intact complement system to lyse sheep erythrocytes when coated with specific antibodies . The bactericidal activity of the serum against Staphylococcus aureus and Escherichia coli was used to evaluate the alternative complement pathway . Sera were obtained from neonates (40 +/- 2 weeks of gestation), and one-month or three-month old infants, fed either breast or formula . Control serum was obtained from healthy adults between 22 and 30 years of age . The hemolytic capacity of serum from breast-fed infants of one month and three months of age was significantly greater than that of the serum from infants which had been fed formula milk.

Protein Eng, 1995 Dec, 8(12), 1275 - 85
An engineered Staphylococcus aureus PC1 beta-lactamase that hydrolyses third-generation cephalosporins; Zawadzke LE et al.; The beta-lactamase from Staphylococcus aureus PC1 has been cloned into an Escherichia coli vector for site-directed mutagenesis and high-level protein expression . A mutant enzyme has been produced in which Ala238 is replaced by a serine, and Ile239 is deleted (A238S:I239del) . The engineered enzyme hydrolyses third-generation cephalosporins substantially more rapidly than the parental enzyme does, while hydrolysis of benzylpenicillin is slower with the mutant than with the wild-type and native enzymes . The mutant beta-lactamase has been crystallized and the structure determined and refined at 2.8 A resolution . The disposition of the beta-strand which forms the side of the active site is altered in comparison with the native S . aureus beta-lactamase structure, widening the active site cleft and providing space to accommodate the bulky side-chains of the third-generation cephalosporins.

Microb Pathog, 1995 Dec, 19(6), 409 - 19
Internalization of Staphylococcus aureus by cultured osteoblasts; Hudson MC et al.; Staphylococcus aureus is the most common cause of both acute and chronic osteomyelitis; however, the pathogenesis of osteomyelitis is poorly understood . We investigated the ability of S . aureus to associate with chick osteoblasts in culture and have demonstrated internalization of bacteria by the osteoblasts . Two strains of S . aureus were examined that were ingested by osteoblasts to different extents, suggesting strain differences in uptake . Initial association of S . aureus strains with osteoblasts was independent of the presence of matrix collagen produced by the osteoblasts . Internalization of bacteria required live osteoblasts, but not live S . aureus, indicating osteoblasts are active in ingesting the organisms . The bacteria were not killed by the osteoblasts, since viable bacteria were cultured several hours after ingestion.

Planta Med, 1995 Dec, 61(6), 497 - 501
Inhibitory effects of bisbenzylisoquinoline alkaloids on induction of proinflammatory cytokines, interleukin-1 and tumor necrosis factor-alpha; Onai N et al.; Bisbenzylisoquinoline alkaloids are known to affect immune responses as well as inflammatory responses, and have been used for the treatment of inflammatory symptoms in China . This study is aimed at elucidating the inhibitory effects of two alkaloids, fangchinoline and isotetrandrine, on the induction of the proinflammatory cytokines, interleukin-1 (IL-1), and tumor necrosis factor-alpha (TNF-alpha), by Staphylococcus aureus Cowan 1 (SAC)-stimulated human peripheral blood mononuclear cells . These two alkaloids inhibited cytokine production in a dose-dependent manner, and they inhibited it by more than 90% at 10 micrograms/ml at every time point examined . Of note was that these two alkaloids appeared to inhibit IL-1 beta production more effectively than IL-1 alpha production . When the levels of cytokine mRNA were measured by semiquantitative RT-PCR, these alkaloids reduced the levels of the mRNAs of IL-1 beta and TNF-alpha, but not that of beta 2-microglobulin, suggesting that these alkaloids may suppress cytokine transcription selectively.

J Antimicrob Chemother, 1995 Dec, 36(6), 941 - 50
Azithromycin induces in vitro a time-dependent increase in the intracellular killing of Staphylococcus aureus by human polymorphonuclear leucocytes without damaging phagocytes; Silvestri M et al.; Despite its clinical efficacy on intracellular pathogens, the in-vitro intracellular antimicrobial activity of azithromycin, has been shown to be absent or lower than expected from the intracellular concentrations reached . To test the possibility that the high intracellular concentrations of the drug could damage phagocytes, the present study evaluated the effects of azithromycin on (a) the intracellular killing of Staphylococcus aureus by human blood neutrophils (PMNs) and (b) the viability and the respiratory burst of PMNs . Using a fluorochrome assay, we assessed the phagocytosis and intracellular killing of S . aureus by PMNs preloaded with azithromycin, or by PMNs unloaded but with the drug in the culture medium . In addition, possible drug-induced damage to PMNs was evaluated measuring: (a) hydrogen peroxide (H2O2) production and (b) the percentages of PMNs dead at the end of the phagocytosis process . Compared to control PMNs without drug, a time-dependent enhancement in the intracellular killing was observed which was statistically significant after 60 min incubation . The increased intracellular killing was higher in suspensions of unloaded PMNs and azithromycin (P < 0.01) that in suspensions of preloaded PMNs (P < 0.05) . This increased intracellular killing was not associated with increased proportions of dead phagocytes, either in preloaded or unloaded PMNs (P < 0.05, each comparison) . Similarly no changes in the production of H2O2 by PMNs were observed in the presence of azithromycin . Thus, azithromycin induces a time-dependent increase in the bactericidal activity of human PMNs, without increasing the phagocyte self-killing or modifying H2O2 production.

Clin Exp Allergy, 1995 Dec, 25(12), 1218 - 27
Upregulation of IgE synthesis by staphylococcal toxic shock syndrome toxin-1 in peripheral blood mononuclear cells from patients with atopic dermatitis; Hofer MF et al.; BACKGROUND: Atopic dermatitis (AD) is a chronic skin disease associated with increased IgE synthesis and colonization with Staphylococcus aureus secreting exotoxins, such as Toxic Shock Syndrome Toxin-1 (TSST-1) . OBJECTIVES: In this study, we were interested in determining the in vitro effects of TSST-1 on IgE synthesis in peripheral blood mononuclear cells from patients with AD . METHODS: We stimulated peripheral blood mononuclear cells (PBMC) from AD patients with a wide range of TSST-1 concentrations and measured IgE synthesis by enzyme-linked immunosorbent assay (ELISA) after 14 days . RESULTS: We show herein that TSST-1 produced antagonistic effects on IgE synthesis by PBMC from AD patients, depending on the concentration used: IgE synthesis was inhibited at 1000 pg/mL (P < 0.05) and enhanced at 0.01 pg/mL (P < 0.01) of toxin . TSST-1 was found to induce the production of much higher amounts of interferon-gamma (IFN gamma) at 1000 pg/mL than at 0.01 pg/mL of toxin (P = 0.0001) . More importantly, immunoglobulin E (IgE) synthesis was enhanced by TSST-1 at 1 pg/mL in the presence of antibodies blocking IFN gamma activity . The other immunoglobulin (Ig) isotypes were also increased after TSST-1 stimulation suggesting that the enhanced IgE synthesis was secondary to a polyclonal B cell activation rather than an isotype switch . TSST-1-stimulated IgE synthesis was T cell-dependent because purified tonsil B cells were only able to synthesize increased amounts of IgE when small numbers of T cells were added to the cultures . Anti-HLA-DR and anti-LFA-1 monoclonal antibodies (MoAb) inhibited TSST-1-enhanced IgE synthesis, suggesting that the bridging of the T cell receptor (TCR) and major histocompatibility complex (MHC) class II on B cells was necessary for activation of B cell differentiation . CONCLUSION: These data indicate that staphylococcal superantigens are able, at concentrations inducing low amounts of IFN gamma, to stimulate IgE synthesis by PBMC from AD patients, and suggest that staphylococcal toxins may contribute to elevated IgE synthesis in AD.

Jpn J Pharmacol, 1995 Dec, 69(4), 443 - 5
Effect of levofloxacin-albumin Dacron graft on graft infection; Fujimoto K et al.; The susceptibility of Dacron grafts to infection is compared with Dacron grafts applied with levofloxacin-bonded albumin (LVFX-ALB) following the inoculation of 10(7) cells Staphylococcus aureus in rats . Staphylococcus epidermidis was inoculated in the same manner . While the control grafts were infected at the time of removal, the LVFX-ALB Dacron grafts resisted infection, thus demonstrating their effectiveness.

Diabetes Care, 1995 Dec, 18(12), 1588 - 91
Infected puncture wounds in diabetic and nondiabetic adults; Lavery LA et al.; OBJECTIVE: To evaluate bone and soft tissue pathogens resulting from puncture wounds among diabetic and nondiabetic adults . RESEARCH DESIGN AND METHODS: We used a case-control design to compare bacterial pathogens in diabetic and nondiabetic subjects with foot infections precipitated by puncture injuries . We used ICD-9-CM code E920.8 to identify 77 diabetic and 69 nondiabetic patients admitted to the hospital for infected puncture wounds . We identified surgical bone and soft tissue cultures and number and type of organisms per culture . RESULTS: Nondiabetic subjects had significantly less osteomyelitis (13 vs . 35%, P < 0.01) than diabetic subjects and were infected by fewer organisms . Pseudomonas was the most common cause of osteomyelitis among nondiabetic subjects (P < 0.001) . Staphylococcus aureus was more common in diabetic bone (P < 0.001) and soft tissue (P < 0.001) infections . Polymicrobial osteomyelitis was more common in diabetic subjects . There was a longer delay until diabetic subjects received medical treatment compared with nondiabetic subjects (8.7 vs . 5.3 days, P < 0.002) . Diabetic subjects were more likely to have neuropathy (P < 0.001) and to have sustained their injuries while barefoot (P < 0.006) . CONCLUSIONS: Puncture wounds in diabetic subjects were commonly associated with polymicrobial infections . Pseudomonas was the most common cause of nondiabetic osteomyelitis . These results have implications for differential emergent and chronic treatment of puncture wounds in diabetic versus nondiabetic subjects.

Br J Pharmacol, 1995 Dec, 116(8), 3191 - 8
Role for intracellular platelet-activating factor in the circulatory failure in a model of gram-positive shock; De Kimpe SJ et al.; 1 . This study investigates the effects of two structurally different antagonists of platelet-activating factor (PAF), BN52021 and WEB2086, on the circulatory and renal failure elicited by lipoteichoic acid (LTA) from Staphylococcus aureus (an organism without endotoxin) in anaesthetized rats . 2 . Administration of LTA (10 mg kg-1, i.v.) caused hypotension and vascular hyporeactivity to noradrenaline (1 microgram kg-1, i.v.) WEB2086 (5 mg kg-1, i.v., 20 min before and 150 min after LTA) inhibited the delayed fall in mean arterial blood pressure (at 300 min: 99 +/- 6 mmHg vs . 75 +/- 6 mmHg, P < 0.01) and prevented the decrease in pressor response to noradrenaline (at 300 min: 36 +/- 5 mmHg min vs . 17 +/- 5 mmHg min, P < 0.01) . Surprisingly, BN52021 (20 mg kg-1, i.v., 20 min before and 150 min after LTA) neither prevented the hypotension (74 +/- 6 mmHg) nor the vascular hyporeactivity (21 +/- 5 mmHg min) . However, BN52021 inhibited the hypotension to injections of PAF as well as the circulatory failure elicited by lipopolysaccharides (10 mg kg-1, i.v.) . 3 . LTA caused an increase in plasma concentration of creatinine from 39 +/- 5 microM (sham-operated) to 70 +/- 8 microM and urea from 4.7 +/- 0.1 to 13.1 +/- 1.6 mM . The renal failure elicited by LTA was significantly inhibited by WEB2086 (creatinine: 45 +/- 4 microM and urea: 5.7 +/- 0.7 mM), but not by BN52021 . 4 . The induction of nitric oxide synthase activity in lungs by LTA was attenuated by WEB2086 from 98 +/- 17 to 40 +/- 15 pmol L-citrulline 30 min-1 mg-1 protein (P < 0.01), but not by BN52021 (148 +/- 21 pmol L-citrulline 30 min-1 mg-1 protein) . Similarly, WEB2086, but not BN52021, inhibited the increase in plasma nitrite concentration associated with the delayed circulatory failure caused by LTA . The release of tumour necrosis factor-alpha (TNF-alpha) after injection of LTA was not attenuated by WEB2086 . 5 . The induction of nitrite release by cultured macrophages activated with LTA (10 micrograms ml-1 for 24 h) was inhibited by 74 +/- 4% by WEB2086 (3 x 10(-4) M), but not by BN52021, indicating that only WEB2086 acts on intracellular PAF receptors . 6 . Thus, the intracellular release of PAF contributes to the circulatory and renal failure and induction of nitric oxide synthase elicited by LTA in anaesthetized rats . The difference between the two structurally different PAF antagonists in our septic shock models using either LTA or lipopolysaccharide (LPS), shows the importance of models for Gram-positive sepsis in the elucidation of the pathophysiology of septic shock and for the evaluation of potential drugs.

Indian J Biochem Biophys, 1995 Dec, 32(6), 372 - 7
Designing of peptides with immuno-modulatory properties using protein A as a probe; Ray PK et al.; A series of reports from our laboratory have described the multifarious properties of protein A of Staphylococcus aureus Cowan I, apart from its IgG binding affinity . Original reports regarding its anti-tumor, anti-toxic, anti-carcinogenic and immunomodulatory properties published earlier by the authors have implicated some uniqueness of this bacterial protein . It was conceived that such diversified properties must lie in its specific peptide sequences, rendering it to act and behave as a multipotent "Biological Response Modifier" (BRM) . The high resolution X-ray structure of protein A-Fc complex has been delineated earlier, and has been the foundation of many protein engineering studies . This structure along with the amino acid sequence data of its four repetitive domains provided us the basis for designing an octapeptide . This octapeptide was synthesized by solid phase peptide synthesis considering it as the probable site through which PA binds IgG . This octapeptide (NH2-Gln-Asn-Ala-Phe-Tyr-Glu-Ile-Leu-COOH) is present in the first helical segment of B-domain of protein A, and also is a part of domain D, A and C . This octapeptide has been shown to bind IgG by the immunoblotting technique . The binding affinity of the octapeptide appears to be significantly higher than that of intact protein A, as was revealed by calculation of Ka (association constant) and Kd (dissociation constant) values . This octapeptide might serve as a good immunoadsorbant for IgG and/or immune complexes.

Immunol Cell Biol, 1995 Dec, 73(6), 505 - 10
Characterization of immunorelated peptides to porcidin P1; Alberdi F Jr et al.; Porcidin P1, an antimicrobial peptide purified from the granules of porcine polymorphonuclear neutrophils (PMN) using ultrafiltration and reverse phase high performance liquid chromatography (RP-HPLC), was covalently conjugated to BSA and used to generate monospecific polyclonal ascites . Antibodies raised against porcidin P1 were covalently coupled to an Affi-gel Hz affinity column and used for immunoaffinity chromatography of peptides from porcine PMN cell extract . Eleven immunorelated peptides were eluted from the column from neutrophil cell extracts and purified to homogeneity by HPLC . The molecular weights of the immunorelated peptides were determined by mass spectral analysis and ranged in size from 1.91 to 10.65 kDa . Of the 11 immunorelated peptides which were bound to the affinity column, only six peptides were recognized by the anti-porcidin antibodies after HPLC purification . Three immunoreactive peptides displayed potent antibacterial activity towards Staphylococcus aureus and Escherichia coli, reducing viability by as much as 99.9% (> 3 log reduction in CFU) when 5 mu g/mL of each purified peptide was used . The polyclonal monospecific antibodies also reacted with proteins from ovine and human PMN, illustrating possible structural relationships between small antibacterial peptides from the different species.

Int Angiol, 1995 Dec, 14(4), 381 - 4
The value of quantitative bacteriological investigations in the monitoring of treatment of ischaemic ulcerations of lower legs; Majewski W et al.; The quantitative and qualitative bacteriological investigations of 63 patients were done on ischaemic ulcerations before reconstructive vascular surgery and at 7 day intervals after the operation . Among the isolated bacteria the most common were Gram positive (62.9%), especially Staphylococcus aureus . Amputations due to non-healing ulcers were performed on 8 patients, who had ankle brachial index (ABI) lower than 0.47 . In 55 patients with ABI higher than 0.47 (with the exception of one case) free skin grafts were applied to reduce the time of the ulcers healing . Primary healing of ulcers covered with free-skin grafts was achieved in 44 out of 55 patients (80%) . In 11 patients, were free-skin grafts had failed, ulcerations were healed following the repetition of the free-skin grafts . The healing results of skin grafts statistically were significantly better in the group where the number of bacteria in 1 cm2 of ulceration was lower than 50.000 . The severity of infections in ulcers makes the healing process of skin grafts impossible . Quantitative bacteriology additionally helps in objective evaluation of granulating tissue and facilitates choice of proper skin grafting time . This study has shown the usefulness of quantitative bacteriology for the determination of the severity of infections in ulcers.

Int J Pancreatol, 1995 Dec, 18(3), 265 - 70
CT-guided aspiration of suspected pancreatic infection: bacteriology and clinical outcome; Banks PA et al.; We have performed CT-guided percutaneous needle aspiration in 104 patients with severe pancreatitis strongly suspected of harboring pancreatic infection on the basis of systemic toxicity and CT findings (Balthazar CT grade D or E) . Of these 104 patients, 51 (49%) were documented with pancreatic infection . Gram stain was positive in 54 of 58 infected aspirates, and culture was positive in all 58 . Klebsiella, Escherichia coli, and Staphylococcus aureus were the most frequent organisms . Eighty-six percent of infected processes contained only one organism . Overall, pancreatic infection was documented by GPA within the first 2 wk in approx one-half of patients . There were no complications . The overall rate of infection decreased from 60 (1980-1987) to 34% (1988-1995) (p = 0.011) . This change was caused by a reduction in the rate of infected necrosis from 67 to 32% (p = 0.015) . The overall mortality rate remained at 20% . The mortality of sterile pancreatitis was not different from infected pancreatitis (p = 0.14) . We conclude that GPA is a safe, accurate method of diagnosis of pancreatic infection . The rate of pancreatic infection appears to be decreasing . The overall mortality of severe pancreatitis among patients suspected of harboring pancreatic infection has remained unchanged because of the high mortality associated with both infected necrosis and severe sterile necrosis.

East Afr Med J, 1995 Dec, 72(12), 787 - 90
Sequestrectomy and local muscle flap implantation for chronic osteomyelitis in children and adolescents; Bassey LO et al.; Forty one children and adolescents (mean age 11.9 years) with chronic osteomyelitis that were not previously managed surgically were prospectively studied during a 5-year period in the University of Calabar Teaching Hospital, Calabar, Nigeria . All cases were managed by sequestrectcomy of curretage and local muscle flap implantation into the space so created . Preoperative care in this series included blood transfusion for anaemic patients and augmentin (amoxycillin and clavulanic acid) while definitive antibiotic therapy depended on the antimicrobial sensitivity pattern of Staphylococcus aureus and Pseudomonas species which were the predominant bacterial isolates . Overall, the incidence of wound dehiscence and recurrence of chronic osteomyelitis were low (19.5%), and complete healing of all cases occurred by two years of follow-up . The major complications were joint stiffness, limb shortening and pathological fractures.

Infect Control Hosp Epidemiol, 1995 Dec, 16(12), 717 - 24
Ribotyping of nosocomial methicillin-resistant Staphylococcus aureus isolates from a Canadian hospital; Nath SK et al.; OBJECTIVE: To evaluate the clonality of methicillin-resistant Staphylococcus aureus (MRSA) strains among hospitalized patients . SETTING: University-affiliated, 465-bed tertiary-care teaching hospital with adjacent cancer clinic in Hamilton, Ontario, Canada . DESIGN: Thirty-five colonized and 30 infected patients from January 2, 1992, through August 31, 1993, were investigated retrospectively . Analysis by restriction fragment-length polymorphisms of ribosomal RNA genes (ie, ribotyping) of 103 nosocomial isolates of MRSA from these 65 patients and of 25 selected unrelated strains was completed . Ribotyping results were compared with the phage typing data obtained prospectively during the course of prospective MRSA surveillance . RESULTS: HindIII ribotyping was more discriminating than phage typing when epidemiologically unrelated strains were differentiated by these methods (19 different ribotypes versus 14 page types; P < .005) . Two early index cases were identified . Isolates from the index cases were two different strains, identified by ribotyping analysis as ribotype A (clonal group 1) and ribotype B (clonal group 2), respectively . These two ribotypes were not found when typing the unrelated control strains . Thirty-six colonized and infected patients (55%) had clonal group 1 isolates, and 20 (31%) had clonal group 2 isolates . These two clones emerged in the hospital in January and February 1992 and dominated the entire investigated period . There also were six patients with an additional clonal group (group 4) that emerged and disappeared in the second quarter of 1993 . CONCLUSIONS: This study highlights the utility of ribotyping in investigating nosocomial MRSA . Three MRSA clones caused nosocomial colonization or infection in patients at this hospital . Two of these MRSA clones, once introduced, were maintained among our patients throughout the study period.

Infect Control Hosp Epidemiol, 1995 Dec, 16(12), 686 - 96
Control of methicillin-resistant Staphylococcus aureus at a university hospital: one decade later; Jernigan JA et al.; OBJECTIVE: To investigate the cause of increasing rates of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infection at a university hospital . DESIGN: Review of data collected by prospective hospital wide surveillance regarding rates of nosocomial MRSA colonization and infection . SETTING: A 700-bed university hospital providing primary and tertiary care . PATIENTS: Patients admitted to the hospital between 1986 and 1993 who were found to be infected or colonized with MRSA . MAIN OUTCOME MEASUREMENT: Rates of MRSA infection and colonization . RESULTS: MRSA infection or colonization was identified in 399 patients (0.18%) admitted during the 8-year study . There was no correlation between the annual rates of MRSA and methicillin-sensitive Staphylococcus aureus (MSSA) infections (P = .66) . The frequency of both nosocomial and non-nosocomial cases increased significantly over the last 4 years of the study (P < .001 for trend) . The ratio of patients who had acquired MRSA nosocomially to those admitted who already were infected or colonized decreased significantly during the study period (P = .002 for trend) . There was a significant increase in the frequency of patients with MRSA being transferred from nursing homes and other chronic care facilities (P = .011) . A cost-benefit analysis suggested that surveillance cultures of patients transferred from other healthcare facilities would save between $20,062 and $462,067 and prevent from 8 to 41 nosocomial infections . CONCLUSIONS: An increase in the incidence of nosocomial MRSA infection was associated with an increased frequency of transfer of colonized patients from nursing homes and other hospitals . The lack of correlation between rates of MRSA and MSSA infections suggested that MRSA infections significantly increased the overall rate of staphylococcal infection . Screening cultures of transfer patients from facilities with a high prevalence of MRSA may offer significant benefit by preventing nosocomial infections and reducing patient days spent in isolation.

Mol Immunol, 1995 Dec, 32(17-18), 1399 - 404
Human recombinant CD4 and CD4-derived synthetic peptides agglutinate immunoglobulin-coated latex particles . Evidence that residues 25-28 and 35-38 of human CD4 form two separate immunoglobulin binding sites; Lenert P et al.; It has been shown previously that amino acid residues 21-49 of the first extracellular domain of human CD4 form the core of an immunoglobulin (Ig) binding site . Synthetic peptides of human CD4 that encompass this region also bind Ig and, with higher affinity, antigen/antibody complexes . Synthetic peptides also enhance binding of both monomeric and aggregated Ig to monocytic U937 cells and Staphylococcus aureus Protein A . To better characterize the nature of the Ig binding site on CD4, we tested the ability of human recombinant CD4 (rCD4) to agglutinate polystyrene particles coated with Ig . Evidence is presented that soluble rCD4 and CD4 peptide p21-49 were capable of specific agglutination of polystyrene particles coated with polyclonal Ig of either human or sheep origin . Agglutination could be blocked by soluble human polyclonal IgG or F(ab')2 fragments . Both heparin and sulfated dextrans also inhibit agglutination, suggesting that charged residues on rCD4 played an important role in agglutination mediated by rCD4 or CD4 peptide . Similarly, aurintricarboxylic acid (ATA) also blocked agglutination of Ig-coated particles by rCD4 . Agglutination mapping studies performed using truncated peptides revealed the existence of two discrete, closely related Ig binding sites (residues 25-28 and 35-38).

Anaesthesist, 1995 Dec, 44(12), 869 - 74
{Lethal, non-menstrual toxic shock syndrome associated with Staphylococcus aureus sepsis}; Raumanns J et al.; We report a rare case of non-menstrual toxic shock syndrome (TSS) in the course of Staphylococcus aureus sepsis in a 31-year-old primigravida who developed high fever and severe pulmonary and cardiovascular failure within a few hours at the end of the 29th week of a twin pregnancy . Mechanical ventilation was necessary due to signs of adult respiratory distress syndrome (ARDS) and catecholamines were needed to maintain a somewhat adequate blood pressure . A forceps delivery was performed immediately . Postoperatively, the patient was brought to the intensive care unit (ICU) due to the suspicion of severe septic shock . In addition to the extreme cardiovascular instability and massive disturbance of pulmonary gas exchange, the clinical picture was characterised by a disseminated intravascular coagulopathy (DIC) with marked petechial bleeding and ecchymoses on all extremities . Moreover, a confluent, spotty exanthem of the trunk and extremities could be seen . Despite all therapeutic efforts, the patient died within a few hours after admission to the ICU with signs of multiorgan failure . Post-mortem, multiple staphylococcal abscesses were found in the kidneys, liver, and uterus . Moreover, acute ulcerous endocarditis of the mitral valve and septic myocardial foci with myocarditis were seen . The Staph . aureus strain isolated from the blood cultures was shown to produce TSS toxin 1 (TSST-1) and enterotoxin B . In summary, the clinical picture can be interpreted as severe staphylococcal sepsis complicated by TSS . TSS is a specific type of infectious disease, occurring mainly in young women during the menstrual period (80%-90%), but it has also been reported in non-menstrual cases (10%-20%) . It is characterised by sudden-onset high fever, hypotension, rash, mucosal hyperaemia, and various additional symptoms such as myalgia, vomiting, and diarrhoea . The clinical course depends on the extent of the organ failure due to decreased tissue perfusion during hypotension . Severe cases are accompanied by multiple organ-system failure including impaired renal function, which is reversible in nearly all cases . Respiratory failure ranges from interstitial and alveolar aedema to ARDS in 10% of cases; severe DIC is seen in 10%-15% . Another severe clinical complication is cardiac insufficiency . The etiology of TSS is based on a localized or, rarely, systemic infection with certain Staph . aureus strains that are capable of producing toxins, the most important one being TSST-1 . Staph . aureus strains can also produce various other enterotoxins that may be involved in the pathogenesis of TSS . The pathogenetic importance of the toxins is supported by the antibody titers in TSS patients: more than 80% of healthy adults show high levels of antibody titers, whereas 90% of TSS patients exhibit low levels in the acute phase followed by a significant increase during convalescence . It is not clear whether the toxins cause TSS by a direct effect or by release of mediators due to their function as superantigens . The clinical characteristics of non-menstrual TSS are identical to those of menstrual TSS, but it can occur in many clinical settings in both sexes at any age . Severe clinical courses are more frequent in non-menstrual TSS: the mortality is about 8%-11% in non-menstrual TSS compared to 2%-5% in menstrual TSS . The diagnosis is based mainly on clinical signs and the isolation of toxin-producing Staph . aureus strains . Besides antibiotic therapy, treatment is primarily directed to the correction of hypotension and additional organ-system failure . Other therapeutic measures such as the elimination of toxins by plasma separation or the administration of antibodies or gamma-globulins are subjects of investigation with no general recommendations at this time.

Antimicrob Agents Chemother, 1995 Dec, 39(12), 2832 - 4
Comparative in vitro activities of LY191145, a new glycopeptide, and vancomycin against Staphylococcus aureus and Staphylococcus-infected fibrin clots; Kang SL et al.; Bactericidal activities of LY191145, an investigational glycopeptide, and vancomycin against Staphylococcus aureus were evaluated . Only LY191145 at a concentration 16-fold greater than the MIC was able to achieve 99.9% killing against methicillin-susceptible S . aureus (ATCC 25923; 8.0 h) . Both agents demonstrated 99.9% killing against methicillin-resistant clinical isolate S . aureus MRSA67 over 24 h at concentrations 4-, 8-, and 16-fold greater than the MIC, but bacteria were killed at a more rapid rate by LY191145 (1.63 versus 5.02 h; P < 0.001) . Against strain ATCC 25923- and MRSA67-infected fibrin clots, total reductions by LY191145 and vancomycin over 72 h were not statistically significantly different at a concentration 16 times the MIC (1.12 +/- 0.31 and 1.23 +/- 0.13 and 1.40 +/- 0.17 and 1.36 +/- 0.37 CFU/g; respectively) . Increasing the drug concentration to 50 times the MIC did not alter the values significantly, and there was no statistically significant difference between the two agents . Overall, LY191145 exhibited more rapid bactericidal activity than vancomycin against S . aureus, and a concentration 16-fold greater than the MIC appears to be optimal.

Antimicrob Agents Chemother, 1995 Dec, 39(12), 2650 - 5
Inducible NorA-mediated multidrug resistance in Staphylococcus aureus; Kaatz GW et al.; The NorA protein of Staphylococcus aureus mediates the active efflux of hydrophilic fluoroquinolones from the cell, conferring low-level resistance upon the organism . The protein also is capable of transporting additional structurally diverse compounds, indicating that it has a broad substrate specificity . Increased transcription of the norA gene, leading to a greater quantity of the NorA protein within the cytoplasmic membrane, is felt to be the mechanism by which strains possessing such changes resist fluoroquinolones . S . aureus SA-1199 and its in vivo-selected derivative SA-1199B are fluoroquinolone-susceptible and -resistant isolates, respectively; SA-1199B resists hydrophilic fluoroquinolones via a NorA-mediated mediated mechanism in a constitutive manner . SA-1199-3 is an in vitro-produced derivative of SA-1199 in which NorA-mediated multidrug resistance is expressed inducibly . Compared with organisms exposed to subinhibitory concentrations of a NorA substrate for the first time, preexposure of SA-1199-3 to such a compound followed by growth in the presence of that substrate results in the elimination of a 2- to 6-h period of organism killing that occurs prior to the onset of logarithmic growth . The uptake of radiolabeled fluoroquinolone is markedly reduced by preexposure of SA-1199-3 to NorA substrates: such prior exposure also results in a dramatic increase in RNa transcripts that hybridize with a norA probe . Preexposure of SA-1199 and SA-1199B to such substrates results in small increases or no increases in these transcripts . No sequence differences between SA-1199 and SA-1199-3 within the norA gene or flanking DNA were found . It appears likely that the regulation of norA in SA-1193, which may be effected by one or more genetic loci outside the norA region of the chromosome, differs from that of SA-1199 and SA-1199B.

Antimicrob Agents Chemother, 1995 Dec, 39(12), 2631 - 4
Autolysis of methicillin-resistant Staphylococcus aureus is involved in synergism between imipenem and cefotiam; Matsuda K et al.; Imipenem-induced autolysis and the activity of imipenem plus cefotiam were studied in 16 strains of methicillin-resistant Staphylococcus aureus (MRSA) . The degree of imipenem-induced autolysis and the rate of synergistic action of imipenem plus cefotiam varied among strains and did not correlate with susceptibility to either imipenem or cefotiam . However, the degree of autolysis correlated well with susceptibility to the synergistic action of imipenem plus cefotiam . In methicillin-susceptible S . aureus strains, both imipenem-induced autolysis and the synergistic activity of the combined drugs were less than those observed in MRSA strains . Differences in the degree of autolysis were not due to differences in autolytic enzyme production . The autolysis of imipenem-pretreated MRSA was enhanced further by cefotiam, while treatment of cells in the reverse order did not enhance autolysis . These findings indicate that cell wall impairment in MRSA is caused by exposure to imipenem but not to cefotiam and that this difference in drug actions results in synergism between imipenem and cefotiam . The possible participation of penicillin-binding proteins PBP 2' and PBP4 in the observed effect is discussed.

Kansenshogaku Zasshi, 1995 Dec, 69(12), 1348 - 55
{Effects of the Prevotella intermedia culture filtrate and short-chain fatty acids on human polymorphonuclear neutrophil functions}; Touyama M et al.; Anaerobes, in particular Prevotella spp., are the predominant bacteria present in mixed respiratory infections . To study the virulence factor of Prevotella intermedia, we examined the effects of the culture filtrate of P . intermedia on the bactericidal activity of human polymorphonuclear neutrophils (PMNs) . The culture filtrate significantly inhibited the phagocytic killing and the intracellular killing of Staphylococcus aureus by PMNs (p < 0.05), but the phagocytosis of PMNs was not affected . The boiled culture filtrate also inhibited the phagocytic killing of PMNs (p < 0.05), which suggests that the inhibitory effect may be mediated by heat-stable substances . Succinic and isovaleric acid were mainly detected from the culture filtrate by gas chromatography . The migration of PMNs was significantly inhibited by these short-chain fatty acids (15 mM succinic acid, p < 0.01 and 1 mM isovaleric acid, p < 0.05) . The phagocytic killing of PMNs was significantly inhibited by succinic acid (30 mM, p < 0.05) and not by isovaleric acid . Therefore, these inhibitory effects on human PMN functions may be contributed to the virulence of the mixed infections with anaerobes.

Appl Microbiol Biotechnol, 1995 Dec, 44(1-2), 118 - 25
Hyperproduction of a recombinant fusion protein of Staphylococcus aureus V8 protease in Escherichia coli and its processing by OmpT protease to release an active V8 protease derivative; Yabuta M et al.; The expression of a recombinant fusion protein including Staphylococcus aureus V8 protease was studied by using Escherichia coli as the host strain . When the mature V8 protease was expressed as a fusion protein with a truncated E . coli beta-galactosidase (beta-gal97S4D), we could not obtain a sufficient amount of the enzyme because of the toxicity resulting from the expressed protease activity . Synthesis of V8 protease was increased by constructing a sandwich-type fusion protein consisting of beta-gal97S4D, a V8 protease derivative with the 56 C-terminal amino acids deleted (V8 delta 56) and a truncated aminoglycoside-3'-phosphotransferase . This fusion protein was successfully produced as inactive inclusion bodies . To release the V8 delta 56 protease from the fusion protein, we developed a novel processing method using an endogeneous E . coli OmpT protease, which can recognize the dibasic amino acid residues located in the linker peptides of the fusion protein . After solubilizing the inclusion bodies with urea, the V8 delta 56 protein was automatically released from the fusion protein by the OmpT protease, which was coprecipitated with the inclusion bodies . The V8 delta 56 protease thus obtained showed the same enzymatic activity as that of the native V8 protease . We demonstrate in this study that the N-terminal prepro sequence and the C-terminal repeated sequence of this enzyme are not necessary for its enzymatic activity and protein folding.

Hinyokika Kiyo, 1995 Dec, 41(12), 1015 - 8
{A case of testicular tumor associated with the contralateral undescended testis}; Ueda H et al.; A 36-year-old man visited the hospital complaining of painless swelling of the right scrotal contents . His left scrotal contents were absent in the scrotum . Right inguinal orchiectomy was performed on May 17, 1993 under the diagnosis of testicular tumor . The pathological diagnosis of the tumor was pure seminoma . Abdominal computed tomography (CT) and magnetic resonance imaging (MRI) revealed the enlargement of para-aortic lymph nodes and there was a soft tissue signal in the left inguinal region which seemed to be the left testis in MRI . His serum testosterone level was low and gonadotropin level was high . The left orchiopexy was performed on June 24, 1993, but the left testis was finally removed because of methicillin-resistant Staphylococcus aureus (MRSA) infections . Radiation therapy was done against the metastatic lesion in the retroperitoneal lymph nodes and complete remission was obtained . The left undescended testis pathologically showed atrophy of germ cells but there was no appearance of atypia of Leydig cells or germ cells.

Neth J Med, 1995 Dec, 47(6), 291 - 5
Metastatic staphylococcal lung abscess due to a cutaneous furuncle; Buwalda M et al.; A seemingly trivial infection of the skin can lead to fulminant staphylococcal pneumonia and death . This case history describes the evolution of a fatal Staphylococcus aureus sepsis complicated by the development of multiple lung abscesses in a 17-year-old patient . A pre-existing cutaneous furuncle was the only identifiable cause . Early bacteraemic symptoms are described . Multiple cavitory lesions could be seen on a CAT-scan . The authors would like to stress the importance of early and adequate antibiotic treatment.

APMIS, 1995 Dec, 103(12), 885 - 91
Multiresistance in Staphylococcus spp . blood isolates in Finland with special reference to the distribution of the mecA gene among the Staphylococcus epidermidis isolates . The Finnish Study Group for Antimicrobial Resistance; Hyvarinen J et al.; A total of 570 Staphylococcus spp . blood isolates collected in Finland in 1991 were tested for susceptiblity to oxacillin and 19 additional antimicrobial agents . The Staphylococcus epidermidis isolates were also analyzed for the presence of the mecA gene by the polymerase chain reaction (PCR) . Of the 238 S . epidermidis, 137 (58%) were in vitro identified as methicillin-resistant and 5 (2%) exhibited oxacillin MICs between 1 and 3 micrograms/ml . All these isolates were positive for the mecA gene in PCR as an indication of genetic resistance to methicillin, while none of the remaining 96 S . epidermidis isolates (oxacillin MICs < or = 0.25 microgram/ml) was positive . Multiresistance was observed in 123 (87%) of the 142 mecA-positive S . epidermidis . Of the 332 Staphylococcus aureus isolates, only one (0.3%) was phenotypically resistant to methicillin; the strain was also resistant to three other unrelated classes of antimicrobials . True methicillin resistance of this strain was manifested by the presence of the mecA gene in PCR . Based on these results, multiresistance was still extremely rate among the S . aureus in our country, whereas among the S . epidermidis as many as half of the blood isolates in central hospitals were multiresistant.

Epidemiol Infect, 1995 Dec, 115(3), 419 - 26
Analysis of methicillin-resistant and methicillin-susceptible Staphylococcus aureus by a molecular typing method based on coagulase gene polymorphisms; Kobayashi N et al.; A molecular typing method for Staphylococcus aureus based on coagulase gene polymorphisms (coagulase gene typing) was evaluated by examining a total of 240 isolates which comprised 210 methicillin-resistant S . aureus (MRSA) and 30 methicillin-susceptible S . aureus (MSSA) collected from a single hospital . By AluI restriction enzyme digestion of the PCR-amplified 3'-end region of the coagulase gene including 81-bp repeated units, the MRSA and MSSA isolates examined were divided into 6 and 12 restriction fragment length polymorphism (RFLP) patterns, respectively, whereas five patterns were commonly detected in MRSA and MSSA . MRSA isolates that showed a particular RFLP pattern were considered to be predominant in the hospital . Coagulase typing with type-specific antisera was also performed for all S . aureus isolates for comparison . Coagulase types II and VII were most frequently detected and included isolates with four and five different AluI RFLP patterns, respectively, whereas each of the other coagulase types corresponded to a single RFLP pattern . These results indicated that RFLP typing was more discriminatory than serological typing, for typing S . aureus and demonstrated its utility in epidemiologic investigation of S . aureus infection in hospitals.

J Allergy Clin Immunol, 1995 Dec, 96(6 Pt 2), 1136 - 44
Role of LFA-1/ICAM-1-dependent cell adhesion in CD40-mediated inhibition of anti-IgM antibody-induced B-cell death; Mayumi M et al.; Cross-linking of surface IgM by anti-IgM antibody caused activation-induced cell death of a surface IgM+, IgD+ human B lymphoma cell line, B104 . The dying B104 cells did not show the morphology of apoptosis but did show that of necrosis . However, anti-IgM antibody caused apoptosis of another surface IgM+, IgD+ human B lymphoma cell line, DND-39 . The influx of extracellular Ca2+ was necessary for the cell deaths of B104 and DND-39 caused by anti-IgM antibody . Their cell deaths were inhibited by cyclosporine . The anti-IgM antibody-induced cell death of DND-39, but not that of B104, was prevented by costimulation with anti-CD40 antibody . In human peripheral blood B-cells, anti-IgM antibody inhibited cell cycle transition induced by Staphylococcus aureus Cowan I at the G2/M interphase without inhibition of DNA synthesis . In this system, too, anti-CD40 antibody canceled the inhibitory signal transduced through surface IgM and increased the number of M phase cells . Blocking antibodies against the leukocyte function-associated antigen-I/intercellular adhesion molecule-1 system decreased the rescue effect of anti-CD40 antibody in both DND-39 cells and peripheral B-cells, which shows that leukocyte function-associated antigen-1/intercellular adhesion molecule-1-dependent cell adhesion plays an important role in the CD40-mediated inhibition of surface IgM-mediated negative signals.

Appl Environ Microbiol, 1995 Dec, 61(12), 4389 - 95
Statistical approach for comparison of the growth rates of five strains of Staphylococcus aureus; Dengremont E et al.; The interaction of temperature (10, 14, 25, 31, and 37 degrees C), pH (pH 5, 5.6, 6.5, 7.4, and 8), and NaCl (0, 2, 5, 8, and 10%) in a laboratory medium affects the specific growth of Staphylococcus aureus . From growth curves obtained by the turbidimetric technique, a nonlinear model in which the specific growth rate (mu) is fitted directly, without data transformation and with the residual error variations taken into account, is proposed . This model correctly fits experimental data and gives more biological information than the quadratic polynomial model . Moreover, the comparison of five strains of S . aureus was performed by a principal-component analysis in which the specific growth rate was the identifying characteristic for S . aureus strains . The results obtained from model coefficient comparison among the five strains and from multivariate data analysis allow the same classification of strains to be performed . Two of them have similar behaviors during food spoilage, two others could be distinguished by their capacity to grow at a low temperature, whereas the last one was markedly different from the others.

Gene, 1995 Dec 1, 166(1), 95 - 9
Isolation and characterization of pcp, a gene encoding a pyrrolidone carboxyl peptidase in Staphylococcus aureus; Patti JM et al.; The pcp gene, encoding a pyrrolidone carboxyl peptidase (PYRase), was cloned from a lambda GT11 genomic library prepared from Staphylococcus aureus FDA 574 and sequenced . The pcp gene is located 740 bp downstream from cna, a gene that encodes a collagen-binding adhesin in S . aureus . S . aureus pcp encodes a 212-amino-acid (aa) polypeptide . The pcp gene was overexpressed in Escherichia coli and the PYRase purified to homogeneity . The recombinant enzyme exhibited biological activity, as determined using the chromogenic substrate L-pyroglutamyl-beta-napthylamide . Biochemical analysis of the PYRase using thiol-blocking chemicals suggested that the enzyme belongs to the cysteine peptidase family . Moreover, multiple sequence alignment revealed a high degree of similarity to previously described bacterial PYRases . This family of peptidases has been used to selectively remove the N-terminal pyrrolidone carboxylic acid residue found on certain blocked proteins and peptides prior to aa sequencing . However, the exact biological role of PYRases has yet to be elucidated.

Comput Chem, 1995 Dec, 19(4), 343 - 9
Protein folds and functional similarity; the Greek key/immunoglobulin fold; Crabbe MJ et al.; Investigations on a number of proteins in the Greek key/immunoglobulin superfold group using computer methods suggested that proteins containing modified Greek keys might exhibit some protein-protein interactions similar to those seen with immunoglobulins . The two domain beta-sheet modified Greek key structure of the beta/gamma-crystallins is shared by other proteins, for example the chaperone protein PapD . Based on computer analysis, we tested for protein-protein interactions using protein A from Staphylococcus aureus, and showed that gamma-crystallin, unlike alpha- or gamma s-crystallin, exhibits weak protein A binding . beta-Crystallin exhibits much weaker binding than gamma-crystallin . Protein A interaction may involve hydrophobic interactions around the interconnecting peptide region . We discuss the implications for the molecular evolution of the crystallins, the superfold, and for the molecular interactions in the mammalian lens that could be important in maintaining transparency.

J Thorac Cardiovasc Surg, 1995 Dec, 110(6), 1708 - 20; discussion 1720-4
The prevalence of infective endocarditis after aortic valve replacement; Agnihotri AK et al.; Replacement valve endocarditis occurred in 3.7% of 2443 patients who underwent primary or redo aortic valve replacements at The Prince Charles Hospital between December 31, 1969 and January 1, 1992, based on a cross-sectional follow-up in 1992 which was 98.8% complete . Because some patients had re-replacements during the study period, a total of 2686 operations were considered for analysis . A variety of replacement devices were used, including 571 allografts (21%), 1152 xenografts (43%), and 880 mechanical valves (36%) . Insertion of an allograft valve resulted in a constant risk of endocarditis which, by multivariable hazard function analysis, negated the effect of any early-phase factors (p < 0.0001) . With other replacement devices, the risk of infection peaked early after operation (9 weeks) and then gave way to a constant risk . Compared with the risk associated with allograft valves, constant risk was higher when the replacement device was a Carpentier-Edwards xenograft (n = 1021, p = 0.02) and lower when a St . Jude Medical mechanical valve was used (n = 505, p = 0.05) . In nonallograft recipients, the presence of active preoperative endocarditis (p < 0.0001) or a concomitant synthetic synthetic aortic root replacement (p = 0.0006) increased the magnitude of the early peaking risk . Regardless of replacement device, constant risk was increased in patients with renal dysfunction (p = 0.01), in younger patients 0.04) . When preoperative endocarditis was caused by Staphylococcus aureus, culture-positive postoperative wound infection was associated with increased risk of replacement valve infection (p < 0.001) and when it occurred, the same organism was usually responsible (86%) . Identification of patients at increased risk for replacement valve infection may lead to reduced morbidity through strategies such as selective use of replacement devices and antimicrobial prophylaxis.

J Immunol, 1995 Dec 1, 155(11), 5175 - 83
Inhibition of expression of cyclin A in human B cells by an immunosuppressant mizoribine; Hirohata S et al.; Mizoribine has been used to prevent rejection of organ allografts in humans and in animal models . Recent clinical trials have demonstrated its efficacy in rheumatoid arthritis and lupus nephritis, in which abnormalities of B cell functions are also involved . We therefore examined the effects of mizoribine on the in vitro function of human B cells . IgM production was induced from highly purified B cells obtained from healthy donors by stimulation with Staphylococcus aureus Cowan I (SA) plus IL-2 . Mizoribine suppressed the production of IgM at its pharmacologically attainable concentrations (1 to 3 micrograms/ml) in a dose-dependent manner . Mizoribine had to be present within the first 96 h after the initiation of cultures to exert its suppressive effects on B cell responses . Cell cycle analysis disclosed that mizoribine significantly decreased the numbers of B cells in S + G2 + M phases . Mizoribine did not decrease GTP levels in SA-stimulated B cells, whereas mizoribine led to a decrease in GTP levels in activated T cells, which was reversed by addition of GMP . Consistently, the suppressive effects of mizoribine on the IgM production of SA-stimulated B cells was not reversed by the addition of GMP as much as 40 microM, which overcame the inhibitory effects of mizoribine on the proliferation of anti-CD3-stimulated T cells . Although mizoribine did not suppress the expression of CD25 and cdc2 kinase, mizoribine markedly suppressed the expression of cyclin A in SA-activated B cells . These results indicate that mizoribine directly suppresses the function of human B cells without interfering with the initial phase of activation . Moreover, the data demonstrate that mizoribine interferes with the cell cycle progression of activated B cells by suppressing the expression of cyclin A by a mechanism distinct from guanine ribonucleotide depletion.

Infect Immun, 1995 Dec, 63(12), 4738 - 43
Role of Staphylococcus aureus coagulase and clumping factor in pathogenesis of experimental endocarditis; Moreillon P et al.; The pathogenic role of staphylococcal coagulase and clumping factor was investigated in the rat model of endocarditis . The coagulase-producing and clumping factor-producing parent strain Staphylococcus aureus Newman and a series of mutants defective in either coagulase, clumping factor, or both were tested for their ability (i) to attach in vitro to either rat fibrinogen or platelet-fibrin clots and (ii) to produce endocarditis in rats with catheter-induced aortic vegetations . In vitro, the clumping factor-defective mutants were up to 100 times less able than the wild type strain to attach to fibrinogen and also significantly less adherent than the parents to platelet-fibrin clots . Coagulase-defective mutants, in contrast, were not altered in their in vitro adherence phenotype . The rate of in vivo infection was inoculum dependent . Clumping factor-defective mutants produced ca . 50% less endocarditis than the parent organisms when injected at inoculum sizes infecting, respectively, 40 and 80% (ID40 and ID80, respectively) of rats with the wild-type strain . This was a trend at the ID40 but was statistically significant at the ID80 (P < 0.05) . Coagulase-defective bacteria were not affected in their infectivity . Complementation of a clumping factor-defective mutant with a copy of the wild-type clumping factor gene restored both its in vitro adherence and its in vivo infectivity . These results show that clumping factor plays a specific role in the pathogenesis of S . aureus endocarditis . Nevertheless, the rate of endocarditis with clumping factor-defective mutants increased with larger inocula, indicating the contribution of additional pathogenic determinants in the infective process.

J Acquir Immune Defic Syndr Hum Retrovirol, 1995 Dec 1, 10(4), 393 - 9
Superantigen toxic shock syndrome toxin-1 (TSST-1) enhances the replication of HIV-1 in peripheral blood mononuclear cells through selective activation of CD4+ T lymphocytes; Hashimoto K et al.; Staphylococcus aureus has been recognized as a common cause of bacteremia of such infections in human immunodeficiency virus type 1 (HIV-1)-seropositive patients . Some staphylococcal exotoxins are recognized as superantigens . We have found that superantigen toxic shock syndrome toxin-1 (TSST-1) brings about a high level of viral production in HIV-1-infected peripheral blood mononuclear cells (PBMCs) through their activation in vitro . The p24 antigen level in the culture supernatant markedly increased in the presence of TSST-1 at a concentration of 1 pg/ml or higher . Fluorescent-activated cell sorter analysis revealed that TSST-1 specifically activated CD4+ T lymphocytes . Although significant production of tumor necrosis factor alpha (TNF-alpha) was observed in uninfected PBMCs treated with TSST-1 after 96 h of incubation, much earlier (after 12 h of incubation) production of TNF-alpha was identified in HIV-1 infected PBMCs with or without TSST-1 treatment . The addition of anti-TNF-alpha antibody to the culture medium resulted in a dramatic decrease in HIV-1 replication . These results suggest that the enhanced replication of HIV-1 by TSST-1 in PBMCs is attributable mainly to the activation of CD4+ T lymphocytes and that the induction of TNF-alpha further enhances replication . Since the enhancement of HIV-1 replication by TSST-1 occurs in a concentration range of picograms per milliliter, the superantigen TSST-1 may play an important role in the pathogenesis and clinical course of HIV-1 infections.

J Exp Med, 1995 Dec 1, 182(6), 1673 - 82
Molecules from Staphylococcus aureus that bind CD14 and stimulate innate immune responses; Kusunoki T et al.; Mammals mount a rapid inflammatory response to gram-negative bacteria by recognizing lipopolysaccharide (LPS, endotoxin) . LPS binds to CD14, and the resulting LPS-CD14 complex induces synthesis of cytokines and up-regulation of adhesion molecules in a variety of cell types . Gram-positive bacteria provoke a very similar inflammatory response, but the molecules that provoke innate responses to these bacteria have not been defined . Here we show that protein-free, phenol extracts of Staphylococcus aureus contain a minor component that stimulates adhesion of neutrophils and cytokine production in monocytes and in the astrocytoma cell line, U373 . Responses to this component do not absolutely require CD14, but addition of soluble CD14 enhances sensitivity of U373 cells by up to 100-fold, and blocking CD14 on monocytes decreases sensitivity nearly 1,000-fold . Deletion of residues 57-64 of CD14, which are required for responses to LPS, also eliminates CD14-dependent responses to S . aureus molecules . The stimulatory component of S . aureus binds CD14 and blocks binding of radioactive LPS . Unlike LPS, the activity of S . aureus molecules was neither enhanced by LPS binding protein nor inhibited by bactericidal/permeability increasing protein . The active factor in extracts of S . aureus is also structurally and functionally distinct from the abundant species known as lipoteichoic acid (LTA) . Cell-stimulating activity fractionates differently from LTA on a reverse-phase column, pure LTA fails to stimulate cells, and LTA antagonizes the action of LPS in assays of IL-6 production . These studies suggest that mammals may use CD14 in innate responses to both gram-negative and gram-positive bacteria, and that gram-positive bacteria may contain an apparently unique, CD14-binding species that initiates cellular responses.

Arch Surg, 1995 Dec, 130(12), 1309 - 13
Primary vs secondary iliopsoas abscess . Presentation, microbiology, and treatment; Santaella RO et al.; OBJECTIVE: To review the characteristics of patient presentation, microbiology, and treatment of primary iliopsoas abscess . DESIGN: A case series of patients with iliopsoas abscess diagnosed on computed tomographic scans from 1987 to 1994 . SETTING: Tertiary care inner-city university hospital . PATIENTS: Eleven patients with secondary iliopsoas abscess, defined as being secondary to gastrointestinal or genitourinary causes or trauma, and seven patients with primary abscess, defined as the absence of the above causes . MAIN OUTCOME MEASURES: Patient characteristics, presenting symptoms and signs, microbiologic characteristics, treatment, and clinical course of patients with primary iliopsoas abscesses compared with those in patients with secondary abscesses . RESULTS: In the primary group, six patients (86%) were intravenous drug users and four (57%) were positive for human immunodeficiency virus . Staphylococcus aureus grew from cultures from five of seven patients with primary abscesses, whereas secondary abscesses had enteric flora . The typical patient presentation included fever, with complaints of pain in the flank, hip, or abdomen . Comparison of abscess drainage options showed shorter hospitalizations for surgical drainage than for percutaneous drainage (15.9 vs 28.5 days; P < or = .01) . CONCLUSIONS: A patient who presents with pain in the flank, hip, or abdomen may have a primary iliopsoas abscess . Computed tomography is the standard method of diagnosis . Antibiotic regimens for patients with primary iliopsoas abscess should include coverage for S aureus, and patients with secondary abscesses should have antibiotic regimens tailored for enteric bacteria . Drainage of abscess is essential for appropriate treatment, and surgical drainage is superior to percutaneous drainage in achieving prompt recovery.

J Lab Clin Med, 1995 Dec, 126(6), 548 - 58
Protective effects of reconstituted high-density lipoprotein in rabbit gram-negative bacteremia models; Hubsch AP et al.; Reconstituted high-density lipoproteins (rHDLs) have the ability to bind bacterial lipopolysaccharide and to reduce its endotoxin activity in vitro and in vivo . The aim of the present studies was to investigate the therapeutic potential of rHDL in bacteremia models . Gram-negative sepsis was induced in anesthetized rabbits by intravenous infusion of Escherichia coli organisms (4 x 10(9) CFU/kg infused over 2 hours) and treated with appropriate antibiotics . rHDL or placebo was infused either before (prophylaxis) or 1 hour after (therapy) the beginning of the bacterial challenge . In the control groups, the bacterial challenge resulted in transient bacteremia, high plasma levels of lipopolysaccharide, secretion of TNF, and symptoms of sepsis, including hypotension and acidosis . rHDL had no influence on blood bacterial counts; however, plasma lipopolysaccharide levels were significantly reduced . Peak plasma TNF concentrations were reduced after prophylactic but not after therapeutic rHDL administration . Both prophylactic and therapeutic rHDL improved clinical outcome: acidosis was significantly attenuated and blood pressure tended to be higher in the rHDL groups . No effects of rHDL were seen in a similar model of gram-positive sepsis induced by the infusion of Staphylococcus aureus.

Plast Reconstr Surg, 1995 Dec, 96(7), 1676 - 88
An experimental model to determine the effect of irradiated tissue on neutrophil function; Gabka CJ et al.; Complications of irradiated tissue include infections and impaired healing . Although fibrosis and hypovascularity contribute, a cellular mechanism has not been identified . This study examines the effect of radiation (10 to 30 Gy) on neutrophil function in a rabbit wound cylinder model . At 3 to 12 weeks after radiation, subcutaneous wound cylinders were implanted in both irradiated and control fields in 19 rabbits . Wound neutrophils were subsequently assayed for phagocytosis (3H-labeled Staphylococcus aureus assay), superoxide production (cytochrome c reduction assay), and surface Mac-1 expression (flow cytometric assay using MHM 23 monoclonal antibody) . Phagocytosis of 3H-labeled S . aureus was significantly lower in neutrophils from irradiated fields compared with controls at 6 and 12 weeks after radiation (6.5 versus 18.9 bacteria per neutrophil at 12 weeks; p = 0.027) . Stimulated neutrophils from irradiated tissue could not increase superoxide production or Mac-1 expression as much as controls, with differences increasing as postirradiation time increased . The diminished phagocytosis, superoxide production, and Mac-1 expression provide a cellular mechanism that may account for susceptibility to infection and poor healing in irradiated tissues.

Proc Natl Acad Sci U S A, 1995 Nov 21, 92(24), 11249 - 53
CD27-CD70 interactions regulate B-cell activation by T cells; Kobata T et al.; CD27, a member of the tumor necrosis factor (TNF) receptor family, binds to its ligand CD70, a member of the TNF family, and subsequently induces T-cell costimulation and B-cell activation . CD27 is expressed on resting T and B cells, whereas CD70 is expressed on activated T and B cells . Utilizing transfected murine pre-B-cell lines expressing human CD27 or CD70, we have examined the effect of such transfectant cells on human B-cell IgG production and B-cell proliferation . We show that the addition of CD27-transfected cells to a T-cell-dependent, pokeweed mitogen-driven B-cell IgG synthesis system resulted in marked inhibition of IgG production, whereas the addition of CD70-transfected cells enhanced IgG production . The inhibition and enhancement of pokeweed mitogen-driven IgG production by CD27 and CD70 transfectants were abrogated by pretreatment with anti-CD27 and anti-CD70 monoclonal antibodies, respectively . In contrast, little or no inhibition of IgG production and B-cell proliferation was noted with CD27-transfected cells or either anti-CD27 or CD70 monoclonal antibody in a T-cell-independent Staphylococcus aureus/interleukin 2-driven B-cell activation system . In this same system CD70-transfected cells enhanced B-cell IgG production and B-cell proliferation, and this enhancement could be gradually abrogated by addition of increasing numbers of CD27-transfected cells . These results clearly demonstrate that interactions among subsets of T cells expressing CD27 and CD70 play a key role in regulating B-cell activation and immunoglobulin synthesis.

J Immunol, 1995 Nov 15, 155(10), 5057 - 63
Localization of an Fc-binding reactivity to the constant region of human IgG4 . Implications for the pathogenesis of rheumatoid arthritis; Zack DJ et al.; The majority of plasma cells in rheumatoid arthritis (RA) synovium produce rheumatoid factors (RF) . IgG RF predominate in the immune complexes found in RA synovial fluid and have been implicated in the pathogenesis of RA . IgG4 RF are a major component of IgG RF produced in serum and synovium of RA patients, even though this subclass comprises only 4% of the serum IgG . We produced an IgG4 mAb, hRF-1, with RF reactivity from the synovial tissue of a patient with RA . mAb hRF-1 had binding specificity for mammalian IgG similar to Staphylococcus aureus protein A, which is characteristic of RF from patients with RA . To determine the molecular basis of this particular RF reactivity, the heavy and light chain genes of mAb hRF-1 were amplified by PCR, cloned, and ligated into the pSG5 plasmid for expression in COS-7 cells . Chain recombination experiments localized the Fc-binding reactivity to the hRF-1 heavy chain . Using a series of chimeric Ab sequences, the Fc-binding reactivity was mapped to the constant region of IgG4 rather than the variable region involved in classic RF reactivity . Multiple domains, including Hinge, CH2, and CH3 of the IgG4 constant region were required for Fc binding . Our studies demonstrate an example of RF-like Fc-binding reactivity that is conferred by the gamma-4 constant region rather than the classic Ag binding site and suggest that increased production of IgG4 may contribute to the pathogenesis of RA.

J Immunol, 1995 Nov 15, 155(10), 4757 - 65
Interference of distinct invariant chain regions with superantigen contact area and antigenic peptide binding groove of HLA-DR; Vogt AB et al.; In the endoplasmic reticulum, MHC class II alpha beta dimers associate with the trimeric invariant chain (li), generating a nine-subunit (alpha beta li)3 complex . In the presence of li, the peptide binding groove is blocked, so that loading with self or antigenic peptides can only occur after proteolytic removal of li in specialized post-Golgi compartments . The class II-associated invariant chain peptide region of li (about residues 81-104) is known to mediate binding to class II molecules and blockade of the groove, but this does not exclude additional contact sites for li . Using a set of overlapping li peptides and recombinant soluble li, we demonstrate here that a large segment of li encompassing approximately residues 71 to 128 interacts with HLA-DR molecules . The N- and C-terminal regions of this li segment appear to bind outside the peptide groove to the contact area for the staphylococcal superantigen Staphylococcus aureus enterotoxin B on the alpha 1 domain . The core region of this segment (residues 95-108) prevents binding of antigenic peptides, probably by interaction with the peptide groove . Occupation of the groove with antigenic peptides abolishes binding not only of the core region, but also that of those li peptides that bind outside the groove . These findings suggest the existence of distinct conformational states of class II molecules, with li binding preferentially to one form.

Ned Tijdschr Geneeskd, 1995 Nov 11, 139(45), 2314 - 6
{Gentamicin skin ointment: 200 kilos too much}; Renders NH et al.; In a 75 year old woman, with diabetic mellitus type II, bacteriological examination of a foot infection revealed a Staphylococcus aureus resistant to aminoglycosides . From a different ulcer on the same foot a genotypically identical but non-resistant bacterial strain was cultured . The patient had been treated with gentamicin ointment . According to the literature and to the Central Pharmaceutical Committee in the Netherlands there is no indication for topical use of gentamicin or other aminoglycosides . Moreover, such use may lead to antibiotic resistant bacteria . In spite of this, in the period from March 1992 to March 1993, 200 kg of the ointment was distributed in the Netherlands.

Plasmid, 1995 Nov, 34(3), 198 - 205
The staphylococcal insertion sequence IS257 is active; Needham C et al.; The plasmid pJ3356 confers high-level mupirocin resistance on a strain of Staphylococcus aureus isolated from a hospital . The plasmid also carries two copies of IS257 . Recombination of an IS257-containing plasmid conferring erythromycin resistance, pOX7-IS, into either of the IS257s of pJ3356 has been observed . The co-integration of pJ3356 and a small plasmid, pOX7, is also reported and involves duplication of one of the IS257s from pJ3356 together with 8 bp of pOX7 at the site of integration . Thus IS257 has been shown to be an active mobile genetic element.

Can J Physiol Pharmacol, 1995 Nov, 73(11), 1669 - 73
alpha-Toxin perfusion: a new method for selective impairment of endothelial function in isolated vessels or intact vascular beds; Laher I et al.; We describe a method for selectively permeabilizing endothelial cells, using the membrane pore forming exoprotein Staphylococcus aureus alpha-toxin . Experiments were performed in rabbit central ear artery or its main side branch under isometric conditions, on the isolated perfused kidney, or in cannulated pressurized renal arteries . In presence of alpha-toxin, endothelial-dependent vasodilator responses elicited by acetylcholine or A23187 were abolished, whereas the sensitivity of smooth muscle cells to constrictors (norepinephrine, phenylephrine, or KCl) or dilators (sodium nitroprusside) was not affected . The results indicate that restricting the alpha-toxin to the luminal surface induces selective impairment of vascular endothelial function . This method of eliminating endothelium-dependent vasodilator responses may prove to be useful in the study of endothelial-smooth muscle interactions of isolated small arteries and intact vascular beds.

Acta Med Port, 1995 Nov, 8(11), 643 - 7
{Spinal epidural empyemas}; Evangelista T et al.; We report ten patients harbouring spinal epidural abscess, aiming to evaluate the factors that may lead to an early diagnosis and that can eventually influence the prognosis . There were seven males and 3 females, with ages comprised between 17 and 66 years . Abscesses were localised mainly in the dorsal region . The most important predisposing factors were infections or other disorders know to be related with compromise of the immunological system . Back pain with or without signs of spinal cord involvement was the most frequent clinical presentation . The pretreatment average time was 16,3 days . Staphylococcus aureus was the most commun organism isolated . Erythrocyte sedimentation rate (ERS) was uniformly elevated, being the most important laboratory data for the diagnosis of this situation . Diagnosis was frequently made on clinical grounds but it was always confirmed by myelography with computed tomography or magnetic resonance . All patients were submitted to surgical drainage . Two patients recovered totally, 5 partially, 2 did not recover at all and 1 died . We conclude that the prognosis is related to the surgical delay and that it depends on the identification of the predisposing factors and the recognition of a clinical picture of spinal cord involvement associated to an elevated ESR . Magnetic resonance is the most reliable imaging technique.

Kansenshogaku Zasshi, 1995 Nov, 69(11), 1272 - 7
{Clinical utility of DNA fingerprinting by arbitrarily-primed polymerase chain reaction (AP-PCR) in nosocomial infection caused by methicillin-resistant Staphylococcus aureus}; Hojo S et al.; Recently, nosocomial outbreaks of MRSA have become an serious social problem in Japan . To examine the routes of transmission of MRSA, the establishment of an accurate MRSA typing system is essential . Previously, we reported that the DNA fingerprinting by AP-PCR might be a useful method to differentiate MRSA strains . In this study, we tried to investigate the clinical usefulness of DNA fingerprinting by AP-PCR using clinically-isolated MRSA strains . Twenty-four MRSA strains (12 with coagulase type IV, and 12 with coagulase type II) isolated from patients in our department were used . Other typing methods (the sensitivity of antibiotics, pulsed-field gel electrophoresis, and plasmid analysis) were also performed . As a result, the typing pattern by AP-PCR correlated well with other typing methods . MRSA strains with coagulase type IV showed almost the same pattern, suggesting that these strains were nosocomially transmitted . On the other hand, MRSA strains with coagulase type II showed various patterns, suggesting these strains were not nosocomially transmitted . In conclusion, the typing by AP-PCR seemed to be a useful tool evaluate a nosocomial MRSA transmission.

Kansenshogaku Zasshi, 1995 Nov, 69(11), 1260 - 8
{A clinical study of gastrointestinal flora in patients with, or without MRSA colonization in the upper-respiratory tract after introduction of preventive measures of hospital infection}; Masaki H et al.; In early 1980's methicillin-resistant Staphylococcus aureus (MRSA) was reported as a major pathogenic organism of geriatric hospital infection in Japan . At the same time in the A geriatric hospital MRSA infection was prevalent . To decrease nosocomial infections some active preventive measures against hospital infection were taken since Oct . 1991 . After introduction of preventive measures of hospital infection in geriatric ward (190 beds) nosocomial bacteremia and pneumonia were markedly decreased in comparison to the episode number before introduction of prevention . However several patients with MRSA colonization were observed every month . The aim of this clinical study was to clear how frequent MRSA was isolated from the gastric juice and stool . Any MRSA was not observed in 63 cultured stool, but just one MRSA was isolated in patients with MRSA colonization . On the other hand gram-negative organisms, which were E . coli, P . aeruginosa, P . mirabilis etc., were frequently observed in cultured stool . In conclusion, we considered frequency of MRSA colonization in gastrointestinal space was not so high but rather very low.

Kansenshogaku Zasshi, 1995 Nov, 69(11), 1235 - 43
{Effects of antiseptics against methicillin-resistant Staphylococcus aureus}; Watanabe Y et al.; Various antiseptics, commonly used in hospital, were tested against methicillin-resistant Staphylococcus aureus (MRSA) at high concentrations (approximately 10(9) CFU/ml) and in short time exposures . The antiseptic solutions of 0.05% alkyldiaminoethylglycine hydrochloride, 0.05% benzalkonium chloride, 0.2% povidone-iodine gargle and 0.03% dominophen bromide produced 1,000 to 1/100,000 reduction after a 30 sec exposure . 7.5 % povidone-iodine scrub and 0.2% benzalkonium chloride in 83% ethanol were most effective, reducing MRSA under detection limits . (less than 10 CFU/ml) 0.05% chlorhexidine gluconate was less effective, producing no evident reduction even after 1 min of exposure . Though 0.2% povidone solution reduced MRSA under detection limits at 37, 24 hrs incubation, colonies of several strains were detected after 48 hrs extended incubation . This suggests the existence of strains resistant to the antiseptic solution of povidone-iodine . We recommend that the disinfection for these strains should be repeated short time disinfection, because repeated short time exposure (15 sec 2 times) of antiseptics was more effective than one long time exposure (30 sec once).

Bioorg Khim, 1995 Nov, 21(11), 845 - 54
{Design of recombinant Escherichia coli strains, determining the secretory expression of artificial human granulocyte-macrophage colony-stimulating factor genes}; Petrovskaia LE et al.; A number of recombinant plasmids for expression of artificial genes encoding human granulocyte-macrophage colony stimulating factor (GM-CSF) were constructed . A hybrid gene was obtained that contains a sequence encoding the leader peptide and a tandem of two IgG-binding domains of protein A from Staphylococcus aureus coupled, through an enteropepdidase linker, to a synthetic gmcsf gene . The construction enables Escherichia coli to carry out biosynthesis of the hybrid protein and its subsequent transport into the periplasmic space of bacteria . Another hybrid gene, combining sequences for the signal peptide of the E.coli outer membrane protein OmpA and GM-CSF, was obtained using polymerase chain reaction . The localization of the mature protein produced by the hybrid gene was found to depend on the strength of the promoter used.

J Antimicrob Chemother, 1995 Nov, 36(5), 851 - 5
The effect of burn wound surgery and teicoplanin on the bactericidal activity of polymorphonuclear leucocytes against Staphylococcus aureus; Steer JA et al.; Polymorphonuclear leucocyte (PMN) function is suppressed for several weeks after burn injury, rendering patients susceptible to infection, commonly with Staphylococcus aureus . A study was performed to determine the effects of surgery to the burn wound and antimicrobial prophylaxis with teicoplanin on killing of S . aureus by PMNs taken from burn patients . The bactericidal rate was significantly reduced before surgery compared to controls (P < 0.01 Mann Whitney), but neither surgery nor teicoplanin had any significant additional effect on the bactericidal rate . The bactericidal rate of PMNs did not have any significant effect on clinical outcome following surgery.

Eur Respir J, 1995 Nov, 8(11), 1934 - 9
Parapneumonic effusions secondary to community-acquired bacterial pneumonia in human immunodeficiency virus-infected patients; Gil Suay V et al.; The purpose of this study was to determine whether the clinical and microbiological characteristics of parapneumonic effusions in patients with community-acquired pneumonia (CAP) infected with the human immunodeficiency virus (HIV) were different from those observed in patients without HIV infection . One hundred and thirty seven patients with parapneumonic effusions were included and divided into two groups depending on whether they had HIV infection or not . The parapneumonic effusion rate was significantly higher in HIV-positive than in noninfected patients (21 vs 13%) . Their clinical course was more severe, presenting a higher rate of bacteraemias (58 vs 18%) . Pleural fluid in patients infected with HIV had significantly lower glucose levels than that of patients without HIV infection . Chest tube drainage was more frequent in parapneumonic effusions of patients infected with HIV than in those without HIV infection (71 vs 44%) . Staphylococcus aureus was the most common microorganism found in the bacteriological samples of patients with CAP infected with HIV (53 vs 12%) . We conclude that patients with community-acquired pneumonia and HIV infection have a higher rate of parapneumonic effusions and a more severe clinical course than non-HIV patients, and that Staphylococcus aureus predominates in their bacteriological samples.

Biol Pharm Bull, 1995 Nov, 18(11), 1482 - 6
A mechanism of resistance to partial macrolide and streptogramin B antibiotics in Staphylococcus aureus clinically isolated in Hungary; Matsuoka M et al.; A plasmid pEP2104 originated from Staphylococcus aureus was clinically isolated in Hungary during 1977 . The plasmid mediates inducible resistance to PMS-antibiotics; partial macrolide {the 14-membered macrolides, erythromycin (EM) and oleandomycin and the 16-membered macrolides mycinamicin I (MCM I) and mycinamicin II (MCM II)and type B streptogramin (MKM-B) antibiotics . The sequence of 31 amino acid residues obtained by N-terminal analysis of the 63kDa protein (MsrSA) present in the membrane from 8325(pEP2104) cells whose PMS-resistance was induced by a concentration of 1.35 micrograms EM/ml {EM-induced 8325(pEP2104)}, was identical to the corresponding sequence in a membrane protein MsrA related to promoting efflux of {14C}EM {Ross J.I., et al., Mol . Microbiol., 4, 1207 (1990)} . A constitutive PMS-resistant strain 8325(pMC38) was obtained from the 8325( pEP2104) strain in the presence of 1 microgram MCMI/ml . No inactivation of EM in EM-induced 8325(pEP2104) was observed . Moreover, poly (A)-directed polylysine synthesis by a cell-free system containing ribosomes from EM-induced 8325 (pEP2104) cells and S100 from Escherichia coli was inhibited by not only EM but spiramycin and MKM-B {Matsuoka M., et al., Biol . Pharm . Bull., 16, 1288 (1993)} . In addition, ribosomes from both EM-induced 8325 (pEP2104) and 8325(pMC38) strains showed about the same affinity as those from the host stain . NCTC8325 . These results suggest, that like MsrA protein, active drug-efflux due to MsrSA protein may be responsible for PMS-resistance . How can the 8325 (pMC38) strain discriminate PMS-antibiotics from most of 16-membered macrolides and lincosamides? A possible explanation is discussed in terms of the pKa-value related to the physicochemical nature of the antibiotics.

Biomaterials, 1995 Nov, 16(16), 1273 - 7
Infection adjacent to titanium and bone cement implants: an experimental study in rabbits; Sanzen L et al.; A pure titanium cylinder or a piece of bone cement was implanted in each upper tibial metaphysis of 20 rabbits . After 4 months radiograms were taken and 10(4), 10(6), or 10(8) Staphylococcus aureus were injected into each leg through central holes in the implants in three groups of animals . Four weeks later new radiograms, bacteriological and histological biopsies were obtained . Three animals died before the end of the experiment . In animals which received 10(6) or 10(8) S . aureus radiographic signs of infection were found in 11/22 legs with both titanium and bone cement implants . No radiolucent zones developed around the implants . Bacteriological cultures from bone close to the implants were negative in all legs with titanium implants and positive in four legs with cement implants . Seven cultures were negative in spite of radiographic changes . It is concluded that after a proper time for wound healing the bone around unloaded implants of both titanium and bone cement is fairly resistant to infection . In some cases, healing of an infection in the surrounding bone seems possible.

Clin Infect Dis, 1995 Nov, 21(5), 1308 - 12
Methicillin-resistant Staphylococcus aureus as a community organism; Moreno F et al.; An increase in methicillin-resistant Staphylococcus aureus (MRSA) infections prompted a study of MRSA during a 21-month period in a 600-bed university hospital in southern Texas . MRSA cases were classified as community, nosocomial, or transfer cases . A case-control study of risk factors for community MRSA compared with community methicillin-susceptible S . aureus (MSSA) was performed . Pulsed field gel electrophoresis (PFGE) of whole cell DNA typing was used as a marker of strain identity for 31 consecutive isolates collected during the last 8 months of the study . During the 21 months there were 170 patients with MRSA infection or colonization, an incidence of 0.2 per 1,000 patient-days . Ninety-nine (58%) of 170 isolates were from community cases; the community to nosocomial case ratio was 2:1 . No significant risk factors differentiated patients with community MRSA compared with community MSSA . Most community MRSA isolates studied (15 {68%} of 22) had distinct PFGE patterns, as did many nosocomial MRSA isolates (4 {44%} of 9) . MRSA isolates were commonly present on admission to the hospital, and multiple MRSA strains were demonstrated among both community and hospital isolates.

J Hosp Infect, 1995 Nov, 31(3), 225 - 8
Efficacy of gentian violet in the eradication of methicillin-resistant Staphylococcus aureus from skin lesions; Saji M et al.; The efficacy of gentian violet (Gv) in eradicating methicillin-resistant Staphylococcus aureus (MRSA) in decubitus ulcers was investigated . Decubitus ulcers (a total of 18 cases) were scrubbed with Gv aqueous solution 0.1% and ointment containing Gv 0.1% was applied daily . MRSA was not detected in these lesions for 3-34 days (average, 10.5 +/- 2.5 days) after the application of Gv ointment . Before this trial, all patients were treated with povidone-iodine and antibiotics; however, those treatments were not effective in eradicating MRSA from skin lesions . Skin irritation and other systemic side effects caused by Gv were not observed . Our data suggest that Gv is a useful agent for treatment of the decubitus ulcers infected with MRSA.

Antimicrob Agents Chemother, 1995 Nov, 39(11), 2415 - 22
Staphylococcus aureus penicillin-binding protein 4 and intrinsic beta-lactam resistance; Henze UU et al.; Increased levels of production of penicillin-binding protein PBP 4 correlated with in vitro acquired intrinsic beta-lactam resistance in a mutant derived from a susceptible strain of Staphylococcus aureus, strain SG511 Berlin . Truncation of the PBP 4 C-terminal membrane anchor abolished the PBP 4 content of cell membrane preparations as well as the resistance phenotype . A single nucleotide change and a 90-nucleotide deletion, comprising a 14-nucleotide inverted repeat in the noncoding pbp4 gene promoter proximal region, were the only sequence differences between the resistant mutant and the susceptible parent . These mutations were thought to be responsible for the observed overproduction of PBP 4 in the intrinsically beta-lactam-resistant mutant . The pbp4 gene was flanked upstream by the open reading frame abcA, coding for an ATP-binding cassette transporter-like protein showing similarities to eukaryotic multidrug transporters and downstream by a glycerol 3-phosphate cytidyltransferase (tagD)-like open reading frame presumably involved in teichoic acid synthesis . The abcA-pbp4-tagD gene cluster was located in the SmaI-D fragment in the S . aureus 8325 chromosome in close proximity to the RNA polymerase gene rpoB.

Nihon Kyobu Shikkan Gakkai Zasshi, 1995 Nov, 33(11), 1233 - 9
{Effect of inhaled vancomycin hydrochloride on elimination of methicillin-resistant Staphylococcus aureus}; Shirai M et al.; Vancomycin hydrochloride (VCM) is one of the most useful drugs in treating methicillin-resistant Staphylococcus aureus (MRSA) infections . Intravenous administration of the drug is, however, not appropriate in patients with impaired renal or liver function . Thus, we studied the effect of inhaled VCM on MRSA . Fifty-one patients with MRSA in their sputum (35 men and 16 women 21, in the "infected" group and 30 in the "colonized" group, mean age 76.4 years) were studied . MRSA was eliminated in 84.3% of the patients (43 of 51 patients), and the average time required for elimination was 14.7 days . MRSA colonization or infection recurred in 46.5% of the patients (20 of 43 patients), and the duration from elimination until recurrence of MRSA averaged 28 days . Eight patients in whom MRSA was not eliminated by inhaled VCM were not clinically distinguishable from other patients, but their performance status was worse . Two hours after VCM was inhaled, serum VCM concentrations were unmeasurable in 7 patients . The VCM level in sputum peaked at 262.5 micrograms/g just after inhalation, and then gradually decreased . No side effects of this treatment were observed . These results suggest that inhaled VCM can be used to eliminate MRSA.

J Burn Care Rehabil, 1995 Nov-Dec, 16(6), 616 - 21
Comparison of hemodynamic changes resulting from toxic shock syndrome toxin-1-producing Staphylococcus aureus sepsis and endotoxin-producing gram-negative rod sepsis in patients with severe burns; Tanaka H et al.; Five patients with burns and toxic shock syndrome toxin-1 (TSST-1)-producing Staphylococcus aureus sepsis (TSS group) were treated in a 5-year period at Kyorin University Hospital's Traumatology and Critical Care Center Burn Unit . Hemodynamic and metabolic differences in these patients were compared retrospectively with those in another five patients who were matched by burn index and age and in whom endotoxin-producing gram-negative rod sepsis developed (End group) . Both groups showed hypermetabolic and hyperdynamic changes at the point sepsis developed . There were no significant differences between the two groups in any parameter . At the point septic shock developed, the TSS group showed significantly lower mean (+/- SD) arterial pressure (TSS vs End group, 64 +/- 5 vs 74 +/- 5 mm Hg; p < 0.05), significantly lower systemic vascular resistance index (TSS vs End group, 579 +/- 62 vs 729 +/- 75 dynes.sec.cm-5/m2; p < 0.05), and higher oxygen consumption (TSS vs End group, 190 +/- 7 vs 163 +/- 11 L/min/m2; p < 0.05) compared with the End group . This is the first clinical report that asserts that TSST-1-producing gram-positive sepsis may result in more hypermetabolic and hyperdynamic differences than does endotoxin-producing gram-negative rod septic shock . These responses may indicate a stronger stimulation of cytokine and nitrous oxide synthetic activity by TSST-1 than by endotoxin.

Cytometry, 1995 Nov 1, 21(3), 241 - 7
Application of flow cytometry in toxinology: pathophysiology of human polymorphonuclear leukocytes damaged by a pore-forming toxin from Staphylococcus aureus; Meunier O et al.; The pore-forming activity of leukocidin (PVL) secreted by Staphylococcus aureus has been investigated on human white cells by flow cytometry techniques . This two-component toxin induced morphological modifications of neutrophils and monocytes as detected by forward light scattering measurements, but was inactive on lymphocytes . These modifications were the consequence of pore formation through the cell membrane leading to its permeabilization . In the absence of calcium, PVL formed pores large enough to allow ethidium ions to penetrate the cells and become fluorescent by intercalating nucleic acids . In the presence of calcium, the pores were too small for ethidium entry but allowed an influx of calcium as shown by the increase in fluorescence of Fluo-3 loaded in the cells . This increase in intracellular calcium concentration induced the activation of neutrophils by PVL as shown by the liberation of their granule content measured by a decrease in side light scattering . Furthermore, ethidium fluorescence was used to discriminate the cells sensitive to PVL, and the analysis of differentiated HL-60 cells and cells obtained from a case of chronic myeloid leukemia led to the conclusion that myeloid cells become sensitive to PVL during differentiation to the metamyelocyte stage.

Pediatr Radiol, 1995 Nov, 25 Suppl 1, S205 - 6
CT of staphylococcal anterior mediastinal abscess in an infant; Bungay HK et al.; A 6-week-old girl presented with bronchiolitis secondary to respiratory syncytial virus . Eight days after admission she developed a Staphylococcus aureus infection at a previous intravenous cannula site . Despite antibiotic therapy this led to an anterior mediastinal staphylococcal abscess, which was drained surgically and the patient recovered . Mediastinal abscesses are rare in children: haematogenous spread of infection is an unusual aetiological factor and we believe this to be the first case reported due to an infected cannula site.

Eur J Nucl Med, 1995 Nov, 22(11), 1249 - 55
Specific targeting of infectious foci with radioiodinated human recombinant interleukin-1 in an experimental model; van der Laken CJ et al.; In the present study, radioiodinated human recombinant interleukin-1 (IL-1) was investigated for its potential to image infectious foci in vivo in an animal model of infection . Twenty-four hours after induction of a Staphylococcus aureus abscess in the left calf muscle, mice were i.v . injected with both iodine-125 labelled IL-1 and iodine-131 labelled myoglobin, a size-matched control agent . The animals were killed for tissue biodistribution studies at 2, 6, 12, 24 and 48 h p.i . Gamma camera images were obtained at 6, 24 and 48 h after injecting mice with 123I-IL-1 . Radioiodinated IL-1 rapidly cleared from the body; after 12 h the abscess was the organ with the highest activity . The absolute abscess uptake of 125I-IL-1 remained high compared to 131I-myoglobin, resulting in significantly higher abscess-to-muscle ratios of 125I-IL-1 compared to 131I-myoglobin . The ratios of 125I-IL-1 reached the ultimate value of 44.4+/-10.8 at 48 h p.i., whereas the ratios of 131I-myoglobin did not exceed 5.9+/-0.7 . Gamma camera imaging revealed clearly visible abscesses . In conclusion, our results demonstrate specific retention of radioiodinated IL-1 in the abscess, presumably by interaction of IL-1 with its receptor on the inflammatory cells . The high target-to-background ratios that were obtained over the course of time indicate that the IL-1 receptor may be a valuable target for the imaging of infectious foci.

J Dairy Res, 1995 Nov, 62(4), 577 - 86
Effects of Staphylococcus aureus products on growth and function of bovine mammary myoepithelial cells in vitro; Zavizion B et al.; The effects of culture supernatants conditioned by the growth of Staphylococcus aureus M60 on in vitro growth and functional properties of bovine mammary myoepithelial cells were examined . Myoepithelial cell proliferation was reduced by Staph . aureus M60 culture supernatants . Exposure of myoepithelial cells to culture supernatants of isogeneic mutants of Staph . aureus M60 that produced either alpha or beta toxins reduced proliferation, but to a lesser extent than supernatants from the wild type strain . Thus, alpha and beta toxins may play some role in affecting myoepithelial cell proliferation . Of the cells tested, 42% contracted following addition of oxytocin (10(-7) M) in the culture medium . Treatment of myoepithelial cells for 15 min with Staph . aureus M60 supernatants, prior to addition of oxytocin in the culture medium, increased the number of cells that contracted to 92% . Exposure of cells for 3 h to the same supernatant, prior to addition of oxytocin in the culture medium, abolished oxytocin responsiveness, had no effect on immunolocalization of actin and vimentin, but affected the localization of alpha-actinin within myoepithelial cells . Treatment of myoepithelial cells for 3 h with a combination of purified staphylococcal proteinases XVI and XVII-B abolished oxytocin responsiveness and mimicked the effect of the Staph . aureus culture supernatant . We conclude that Staph . aureus M60 culture supernatant affected proliferation and functional properties of myoepithelial cells.

FEMS Microbiol Lett, 1995 Nov 1, 133(1-2), 9 - 15
Sequence analysis of the region upstream of a peptidoglycan hydrolase-encoding gene from bacteriophage phi 11 of Staphylococcus aureus; Weerakoon LK et al.; The nucleotide sequence of a 1.1-kb DNA fragment upstream of a peptidoglycan hydrolase-encoding gene (lytA) from bacteriophage phi 11 of Staphylococcus aureus was determined to see if the upstream sequences are involved in the transfer of the lytA product through the cytoplasmic membrane . Sequencing revealed three open reading frames of 171, 147 and 435 bp with consensus Shine-Dalgarno sequences located upstream from the ATG start codons . The third open reading frame overlaps with the 5' end of lytA by 18 nucleotides . Comparison of the deduced amino acid sequences of the open reading frames with the amino acid sequences in the NCBI Entrez database did not show any significant homology to any sequenced polypeptides . However, the analysis of the peptides showed some structural similarities to the product of the holin gene family . Lysogens containing an insertional mutation in ORF3, upon induction, produced either no phage titer or very low phage titers, compared to the wild-type lysogen . Transformation of ORF3 mutated lysogens by a plasmid containing the intact ORF3 produced the same phage titer as wild-type lysogen, suggesting that the ORF3 product is involved in the process of cell lysis/phage release.

FEMS Microbiol Lett, 1995 Nov 1, 133(1-2), 155 - 61
Translation of RNAIII, the Staphylococcus aureus agr regulatory RNA molecule, can be activated by a 3'-end deletion; Balaban N et al.; RNAIII, an RNA molecule shown to encode delta-hemolysin and independently to regulate toxin synthesis in Staphylococcus aureus, is transcribed at the mid-exponential phase of growth, while its target genes are activated 2 h later, at the post-exponential phase of growth . We show here that the translation of RNAIII to the 26-amino acid peptide delta-hemolysin is delayed by 1 h, and that this delay is abolished when the 3'-end of this molecule is deleted . We suggest that structural changes of RNAIII to a translatable form of the molecule precede its regulation of target gene expression.

J Antibiot (Tokyo), 1995 Nov, 48(11), 1292 - 8
Two conformers of the glycopeptide antibiotic teicoplanin with distinct ligand binding sites; Westwell MS et al.; The clinically important vancomycin group glycopeptide antibiotics, which act by blocking cell wall synthesis, are crucial in the treatment of methicillin resistant Staphylococcus aureus . All of the group members studied so far, with the apparent exception of teicoplanin, enhance their antibiotic action by the formation of an asymmetric homodimer . Teicoplanin exists in two main conformers which differ by a rotation of approximately 180 degrees of a sugar residue . From NMR studies and molecular modelling, we present structures for the two conformers and conclude that they have different binding affinities for cell wall analogues . The two conformers of teicoplanin are closely analogous to those adopted by each half of the asymmetric dimers of the other vancomycin group antibiotics.

Acta Otorrinolaringol Esp, 1995 Nov-Dec, 46(6), 421 - 6
{Antibiotic prophylaxis in surgery of uncomplicated cholesteatoma: review of 150 patients}; Lopez Amado M et al.; A review was made of 150 cases of chronic otitis media caused by cholesteatoma that were operated consecutively; no complications or active otorrhea was present . In 61.3% of cases, prophylactic antibiotic (PA) treatment was given on the basis of the surgeon's criteria, which probably reflected the presence or absence of risk factors . Most patients were given a penicillin or penicillin derivative (amoxi-ampicillin followed by cephalosporins) . Antibiotic administration was prolonged longer than necessary (mean 3.4 days) . PA was not associated with important complications . Postoperative infection occurred in 10.7% of cases, usually infection of the surgical wound, most commonly by Staphylococcus aureus . Otorrhea occurred in 7.3% of cases in the 15 days after surgery and was more frequent in patients treated via an endomeatal approach and in those who had ossicular prosthesis . Prophylactic antibiotic treatment produced no benefit to our patients in the form of any reduction in number of surgical infections, postoperative otorrhea or mean hospital stay . Therefore, PA is not recommended in surgery for uncomplicated cholesteatoma . When risk factors are present, a suitable and complete antibiotic treatment may be indicated.

Plant Physiol, 1995 Nov, 109(3), 983 - 90
Evidence that spinach leaves express calreticulin but not calsequestrin; Navazio L et al.; The presence of either calreticulin (CR) or calsequestrin (CS-like proteins in spinach (Spinacia oleracea L.) leaves has been previously described . Here we report the purification from spinach leaves of two highly acidic (isoelectric point 5.2) Ca(2+)-binding proteins of 56 and 54 kD by means of DEAE-cellulose chromatography followed by phenyl-Sepharose chromatography in the presence of Zn(2+) (i.e., under experimental conditions that allowed the purification of CR from human liver) . On the other hand, we failed to identify any protein sharing with animal CS the ability to bind to phenyl-Sepharose in the absence of Ca(2+) . Based on the N-terminal amino acid sequence, the 56- and 54-kD spinach Ca(2+)-binding proteins were identified as two distinct isoforms of CR . Therefore, we conclude that CR, and not CS, is expressed in spinach leaves . The 56-kD spinach CR isoform was found to be glycosylated, as judged by ligand blot techniques with concanavalin A and affinity chromatography with concanavalin A-Sepharose . Furthermore, the 56-kD CR was found to differ from rabbit liver CR in amino acid sequence, peptide mapping after partial digestion with Staphylococcus aureus V8 protease, pH-dependent shift of electrophoretic mobility, and immunological cross-reactivity with an antiserum raised to spinach CR, indicating a low degree of structural homology with animal CRs.

Immunology, 1995 Nov, 86(3), 364 - 71
High-level IL-10 production by monoclonal antibody-stimulated human T cells; Schwarz M et al.; We investigated interleukin-10 (IL-10) production in freshly isolated mononuclear cells and purified T cells in response to stimulation with monoclonal antibodies (mAb) recognizing CD3, CD2 and CD28, or with the bacterial products Staphylococcus aureus cells (SAC), staphylococcal enterotoxin (SEA) and lipopolysaccharide (LPS) . IL-10 production was compared with that of IL-2, IL-4 and interferon-gamma (IFN-gamma) . Similar to the other cytokines, in peripheral blood mononuclear cells (PBMC) from adult donors the highest IL-10 levels were produced in response to CD2 plus CD28 stimulation, within 72-96 hr of stimulation . Levels of IL-10 in response to CD2 plus CD28 stimulation (1.9 +/- 1 ng/ml) exceeded those in response to SEA (0.25 +/- 0.16 ng/ml), SAC (0.43 +/- 0.42 ng/ml), or LPS (0.19 +/- 0.14 ng/ml) stimulation . With adult purified T cells, high levels of IL-10 and IL-4 were measured following CD3 plus CD28 stimulation, and the amounts of both T-helper type-2 (Th2) cytokines decreased following the addition of phorbol myristate acetate (PMA), whereas the synthesis of the Th1 cytokines IL-2 and IFN-gamma was enhanced . When PBMC were stimulated with a CD3 mAb and different other cytokines were added, strong enhancement of IL-10 production was seen upon the addition of IL-2, IL-4, IL-7, IL-12 and IFN-gamma, whereas inhibition was found with transforming growth factor-beta 1 (TGF-beta 1) . These data illustrate that in freshly isolated PBMC large amounts of IL-10 can be induced rapidly by appropriate mAb stimulation, and that even in freshly isolated cells IL-4 and IL-10 show signs of parallel regulation.

Immunopharmacol Immunotoxicol, 1995 Nov, 17(4), 759 - 73
Modulation of primary antibody response by protein A in tumor bearing mice; Zaidi SI et al.; Protein A (PA) is a cell wall glycoprotein of Staphylococcus aureus Cowan I, which possess a number of immunomodulatory and antitumor properties . We have previously shown that PA suppresses the anti-sheep erythrocyte primary antibody response in normal mice . The present investigation evaluates the effect of protein A on the anti-sheep erythrocyte primary antibody response in tumor-bearing mice . The primary antibody response in tumor-bearing mice immunized with sheep red blood cells (SRBC) was suppressed by the intraperitoneal administration of PA in a dose-dependent fashion . The plaque forming cell (PFC) assay was used to assess this response . Maximum suppression of the PFC response was observed at 12 micrograms PA/animal (p < 0.001) and could be observed at doses as low as 1 microgram PA/animal (p < 0.01) . The amount of suppression was proportional to the number of PA doses administered . In addition this effect was critically dependent on the timing of PA administration . PA showed no significant effect on PFC when injected after immunization, but it produced pronounced suppression when injected prior to the immunization with SRBC . Maximum suppression of the PFC response was observed when PA was administered one day before the antigen challenge . PA also reduced splenic localization of 51Cr labeled SRBC to 42% (p < 0.01) . The altered localization of antigen in spleen may be responsible for reduced PFC response in tumor-bearing mice . Depletion of B-lymphocyte is reported to exhibit tumor inhibition . Therefore, we propose that the suppression of the primary antibody response by PA helps in tumor regression by reducing the soluble immunosuppressive immune complexes.

HNO, 1995 Nov, 43(11), 664 - 8
{Significance of Staphylococcus aureus in nose operations . Risk of toxic shock syndrome?}; Pennekamp A et al.; In a prospective clinical trial the pre- and postoperative presence of S . aureus was examined in 130 patients undergoing nasal septal surgery . The patients were randomized into three groups . The first group received no perioperative antibiotics, the second group was given oral amoxicillin plus clavulanic acid, while the third group was treated with oral sulfamethoxazol and trimethoprim . A significant decrease in the incidence of S . aureus was observed in post-operative cultures, but the difference was not attributable to the antibiotic use . Overall, 18.9% of the S . aureus carriers harbored toxic shock syndrome toxin-1 positive strains . However, the decrease in the presence of S . aureus and the risk for toxic shock syndrome was not influenced by the antibiotics administered . These findings show that the routine use of oral prophylactic antibiotics for patients undergoing nasal surgery seems not indicated.

Nippon Kyobu Geka Gakkai Zasshi, 1995 Nov, 43(11), 1841 - 4
{Tricuspid valve replacement for infective endocarditis in drug addict--a case report}; Yanagiya A et al.; For right-sided endocarditis associated with drug abuse a successful treatment by tricuspid valve replacement was reported . A 34-year-old female who had a history of intravenous drug use for 14 years was admitted with complaints of chest pain, fever and dyspnea . A large vegetation about 47 mm in size attached to the tricuspid valve with tricuspid regurgitation was detected by echocardiography . Methicillin-resistant staphylococcus aureus was isolated in a blood culture . Because infection was persistent and uncontrollable in spite of sensitive multiple antibiotic regimens, tricuspid valve replacement using a St . Jude Medical valve was successfully performed with excision of markedly destroyed leaflets and debridement of the infectious lesions . After surgery the patient has been free from infection for 3 years.

J Exp Med, 1995 Nov 1, 182(5), 1447 - 59
The interleukin-1 beta-converting enzyme (ICE) is localized on the external cell surface membranes and in the cytoplasmic ground substance of human monocytes by immuno-electron microscopy; Singer II et al.; Interleukin-1 beta (IL-1 beta)-converting enzyme (ICE) is a novel cysteine protease that cleaves the 31-kD inactive cytoplasmic IL-1 beta precursor into active extracellular 17-kD IL-1 beta . The ICE gene product is a 45-kD proenzyme that requires proteolytic processing to activate ICE . Active ICE is a heterodimer consisting of equal amounts of p20 and p10 subunits . Generation of active ICE is affected by the removal of an 11-kD NH2-terminal precursor domain (p11) and an internal 19-amino acid sequence that separates the 20- and 10-kD subunits . Immuno-electron microscopy was performed on human monocytes with immunoglobulins recognizing the active (p20) or precursor (p11) domains of ICE . Elutriated monocytes were stimulated with 50 pM lipopolysaccharide followed by heat-killed Staphylococcus aureus under conditions that induce maximal rates of IL-1 beta secretion . Ultrathin cryosections were cut from fixed frozen pellets of these monocytes and were immunogold labeled with either antibody . Active and precursor domain ICE epitopes were localized in the cytoplasmic ground substance, but they were not detected within the endoplasmic reticulum, the Golgi apparatus, and secretory granules of activated or inactive monocytes . Importantly, numerous ICE p20 epitopes were also observed on the extracellular surfaces of the cell membrane, and were concentrated on the microvilli . Very similar patterns of ICE localization were obtained with unstimulated blood monocytes . In contrast, ICE p11 epitopes were not detected on the surfaces of these monocytes . Likewise, labeling of fixed ultrathin cryosections of monocytes with a biotinylated irreversible ICE inhibitor {Ac-Tyr-Val-Lys(biotin)-Asp-(acyloxy)-methyl-ketone} showed that the compound localized on the outer cell surface as well, and to a lesser extent, within the cytoplasmic ground substance . Furthermore, antipeptide antibodies specific for either the mature or precursor domains of IL-1 beta were both localized upon the cell membrane after stimulation of IL-1 beta secretion . Lipopolysaccaride-primed monocytes that synthesized, but did not secrete IL-1 beta, exhibited only cytoplasmic staining . The data suggests that mature IL-1 beta is generated via cleavage of the 31-kD inactive cytoplasmic IL-1 beta precursor by ICE after association with the plasma membrane during secretion.

J Infect Dis, 1995 Nov, 172(5), 1312 - 6
Systemic prophylaxis of experimental staphylococcal endophthalmitis: comparative efficacy of sparfloxacin, pefloxacin, imipenem, vancomycin, and amikacin; Marrakchi-Benjaafar S et al.; The preventive efficacy of several antibiotics in experimental staphylococcal endophthalmitis was evaluated . Two hours before bilateral intravitreal infection with 500 cfu of Staphylococcus aureus, 18 pigmented phakic rabbits were assigned to receive a single intramuscular injection of sparfloxacin (50 mg/kg), pefloxacin (50 mg/kg), imipenem (50 mg/kg), vancomycin (30 mg/kg), amikacin (15 mg/kg), or saline and were killed 24 h after infection (6 eyes/group) . Sparfloxacin, pefloxacin, and imipenem were significantly (P < .001) more effective than saline . All but 1 of the sparfloxacin-treated eyes were culture negative . To determine whether the effect persisted, an additional 24 rabbits were treated with sparfloxacin, pefloxacin, imipenem, or saline and were killed 48 h after infection (12 eyes/group) . Sparfloxacin, pefloxacin, and imipenem were effective (P < .001) . All sparfloxacin-treated eyes remained culture negative . These results show that systemic antibiotic administration prevents the development of experimental endophthalmitis and that further studies of sparfloxacin as a prophylactic agent are warranted.

J Clin Invest, 1995 Nov, 96(5), 2304 - 10
IL-10 inhibits metalloproteinase and stimulates TIMP-1 production in human mononuclear phagocytes; Lacraz S et al.; Human mononuclear phagocytes can modulate the turnover of extracellular matrix by producing metalloproteinases such as 92-kD gelatinase and interstitial collagenase as well as the tissue inhibitor of metalloproteinases (TIMP) . We have previously reported that IL-4 and IFN gamma released by lymphocytes suppress metalloproteinase biosynthesis in macrophages without affecting TIMP production (Lacraz, S., L . Nicod, B . C . de Rochementeix, C . Baumberger, J . Dayer, and H . Welgus . 1992 . J . Clin . Invest . 90:382-388.; Shapiro, S . D., E . J . Campbell, D . K . Kobayashi, and H . G . Welgus 1990 . J . Clin . Invest . 86:1204-1210) . Like IL-4, IL-10 is secreted by Th2 lymphocytes and is inhibitory to several macrophage functions . In the present study, IL-10 was tested and compared to IL-2, IL-4, IL-6, and IFN gamma for its capacity to modulate synthesis of 92-kD gelatinase, interstitial collagenase and TIMP in human macrophages and monocytes . We found that IL-10, just like IL-4, inhibited the production of 92-kD gelatinase and blocked LPS-, as well as killed Staphylococcus aureus-induced, interstitial collagenase production . The principal finding of this study, however, was that IL-10, in distinction to IL-4, produced a dose-dependent stimulation in the biosynthesis of TIMP-1 . TIMP-2 production was not affected . IL-10 regulated the expression of 92-kD gelatinase and TIMP-1 at the pretranslational level . Furthermore, IL-10 regulation was cell type-specific, as it had no effect on the production of metalloproteinases or TIMP by human fibroblasts . In summary, IL-10 has a potent and unique effect upon tissue macrophages and blood monocytes by enhancing TIMP-1 production while decreasing metalloproteinase biosynthesis.

Invest Ophthalmol Vis Sci, 1995 Nov, 36(12), 2482 - 91
Immunopathologic features of Staphylococcus aureus endophthalmitis in the rat; Ravindranath RM et al.; PURPOSE . To study the clinical, histopathologic, and immunologic responses to Staphylococcus aureus endophthalmitis in rats . METHODS . Experimental Lewis rats received an intravitreal injection of viable S . aureus (65 organisms), and control rats received sterile saline . The clinical scores, cellular infiltrate, delayed hypersensitivity reaction in skin tests, and serum and vitreous enzyme-linked immunosorbent assay titers of immunoglobulin (Ig) M, IgG, and IgA to ribitol teichoic acid (RTA), the major antigenic determinant of S . aureus cell wall, were measured and compared on days 3, 7, 10, 14, 21, and 30 . The differences were statistically assessed using Mann-Whitney nonparametric t-tests and analysis of variance . RESULTS . The red reflex was abolished in the majority of rats between days 3 and 21 . Ocular inflammation resolved by day 30 . The vitreous of eyes injected with S . aureus showed bacterial growth on days 3 and 7, followed by a decrease in numbers on days 10 and 14 and disappearance on days 21 and 30 . In the vitreous, a peak neutrophil count was observed at day 3 that rapidly declined by day 7 . The number of lymphocytes and plasma cells peaked on day 3 but declined more slowly . Plasma cells and Mott cells were seen on days 10 and 14, suggesting intraocular antibody production . IgM titers to RTA increased progressively in serum and vitreous, reached a peak on day 21, and declined on day 30 . A weak IgG but absent IgA response to RTA was observed in serum and vitreous . S . aureus endophthalmitis was not associated with delayed hypersensitivity to the bacteria in skin tests . CONCLUSIONS . S . aureus endophthalmitis is associated with the infiltration of neutrophils, lymphocytes, monocytes, and plasma cells in vitreous . Neutrophils, the predominant infiltrating cells, may be involved in bactericidal activity and opsonophagocytosis . In rat staphylococcal endophthalmitis, IgM rather than IgG may be the protective antibody.

Blood, 1995 Nov 1, 86(9), 3575 - 82
Molecular analysis in three cases of X91- variant chronic granulomatous disease; Bu-Ghanim HN et al.; Defects in gp91-phox, the large subunit of cytochrome b558 (b-245) give rise to X-linked chronic granulomatous disease (CGD), a rare inherited condition characterized by an extreme susceptibility to bacterial and fungal infection . In the majority of cases, the phagocytes are unable to generate any superoxide owing to complete absence of the flavocytochrome . However, a small minority of these patients do have some phagocytic oxidase activity . We describe here an analysis of the molecular basis of the disease in three such variant patients with lesions in the gene coding for gp91-phox on the X chromosome . Three different genetic lesions were found, resulting in the substitution of tyrosine for cysteine 244, a deletion of one of three lysines 313 through 315, and the deletion of the six C-terminal amino acids, respectively . The functional consequences of these defects on oxidase activity was a reduction to 12%, 3.6%, and 2.1% of the normal levels, respectively . Corresponding levels of gp91-phox were 20%, 8%, and 16% of normal classifying these patients as X91- . Microbicidal assays showed that killing of Staphylococcus aureus was grossly impaired in cells in which there was 12% normal activity . This implies that if gene therapy is to be applied, it must restore oxidase activity to a much higher level than that present in the cells of this patient . The sites of two of the mutations were analyzed on a model of the C-terminal half of the gp91-phox, based on the crystal structure of the homologous protein ferrodoxin NADP reductase . Possible structural consequences of the mutations were examined.

J Med Microbiol, 1995 Nov, 43(5), 328 - 34
Antibody response to Staphylococcus aureus collagen binding protein in patients with S . aureus septicaemia and collagen binding properties of corresponding strains; Ryding U et al.; An ELISA was developed for the detection of IgG antibodies to Staphylococcus aureus collagen binding protein (CnBP) in 95 patients with S . aureus endocarditis, complicated septicaemia with bone and joint involvement or uncomplicated septicaemia and in 95 control patients . Sixty percent of S . aureus-infected patients showed a positive peak anti-CnBP antibody level or a significant rise in titre, or both, during infection, but patients with S . aureus endocarditis or complicated septicaemia could not be differentiated from those with uncomplicated S . aureus septicaemia . The collagen binding capacity of S . aureus strains from 82 of the 95 patients was investigated in a particle agglutination assay and a 125I-labelled assay . All strains bound collagen in the particle agglutination assay as did 68% in the 125I-labelled assay, but there was no correlation between collagen binding of the patient strain and endocarditis, joint or skeletal involvement . An anti-CnBP antibody response was seen in 16 patients infected with a S . aureus strain negative for collagen binding in vitro, indicating in-vivo expression of CnBP.

J Med Microbiol, 1995 Nov, 43(5), 318 - 27
Staphylococcal scalded skin syndrome; Gemmell CG; Staphylococcal scalded skin syndrome (SSSS) is a recognised clinical entity that affects primarily the very young and, in rare cases, the very old or the immunocompromised . Koch's postulates have been fulfilled in that: (i) Staphylococcus aureus is isolated from every case; (ii) S . aureus can reproduce the syndrome in an experimental animal model; (iii) a specific extracellular toxin can reproduce the syndrome; and (iv) antibody to the toxin can protect experimental animals . Although exfoliative toxin (ET) is responsible for the skin loosening seen in SSSS, it does not account for all the symptoms of the disease . Purified ET does not cause erythema in either neonatal mice or man, and the lesions are not painful unless the loosened epidermis is removed . This suggests that other factors, e.g., delta-haemolysin, are involved in the pathogenesis of this condition . Although much has been learned about the pathogenesis of the syndrome, we are still largely ignorant of the factors which govern host resistance to SSSS (i.e., intoxication by ET-producing strains of S . aureus) . It is fortunate from the patient's point of view that the aetiological agent can be destroyed readily by the use of appropriate antibiotic therapy.

J Pediatr, 1995 Nov, 127(5), 746 - 8
Cardiac complications in children with Staphylococcus aureus bacteremia; Friedland IR et al.; Clinically silent endocarditis was detected in 4 (11%) of 36 hospitalized children with staphylococcal bacteremia who underwent echocardiographic examination . Pericarditis was detected in two further children . Only one child had underlying cardiac disease (patent ductus arteriosus) . Echocardiography should be considered in children with staphylococcemia even if an obvious extracardiac focus is apparent.

J Biol Chem, 1995 Oct 27, 270(43), 26000 - 5
Effect of the activation of phosphatidylinositol 3-kinase by a thiophosphotyrosine peptide on glucose transport in 3T3-L1 adipocytes; Herbst JJ et al.; Insulin causes the activation of phosphatidylinositol 3-kinase (PI 3-kinase) through complexation of tyrosine-phosphorylated YMXM motifs on insulin receptor substrate 1 with the Src homology 2 domains of PI 3-kinase . Previous studies with inhibitors have indicated that activation of PI 3-kinase is necessary for the stimulation of glucose transport in adipocytes . Here, we investigate whether this activation is sufficient for this effect . Short peptides containing two tyrosine-phosphorylated or thiophosphorylated YMXM motifs potently activated PI 3-kinase in the cytosol from 3T3-L1 adipocytes . Introduction of the phosphatase-resistant thiophosphorylated peptide into 3T3-L1 adipocytes through permeabilization with Staphylococcus aureus alpha-toxin stimulated PI 3-kinase as strongly as insulin . However, under the same conditions the peptide increased glucose transport into the permeabilized cells only 20% as well as insulin . Determination of the distribution of the glucose transporter isotype GLUT4 by confocal immunofluorescence showed that GLUT4 translocation to the plasma membrane can account for the effect of the peptide . These results suggest that one or more other insulin-triggered signaling pathways, besides the PI 3-kinase one, participate in the stimulation of glucose transport.

Proc Natl Acad Sci U S A, 1995 Oct 24, 92(22), 10359 - 63
The cell wall components peptidoglycan and lipoteichoic acid from Staphylococcus aureus act in synergy to cause shock and multiple organ failure; De Kimpe SJ et al.; Although the incidence of Gram-positive sepsis has risen strongly, it is unclear how Gram-positive organisms (without endotoxin) initiate septic shock . We investigated whether two cell wall components from Staphylococcus aureus, peptidoglycan (PepG) and lipoteichoic acid (LTA), can induce the inflammatory response and multiple organ dysfunction syndrome (MODS) associated with septic shock caused by Gram-positive organisms . In cultured macrophages, LTA (10 micrograms/ml), but not PepG (100 micrograms/ml), induces the release of nitric oxide measured as nitrite . PepG, however, caused a 4-fold increase in the production of nitrite elicited by LTA . Furthermore, PepG antibodies inhibited the release of nitrite elicited by killed S . aureus . Administration of both PepG (10 mg/kg; i.v.) and LTA (3 mg/kg; i.v.) in anesthetized rats resulted in the release of tumor necrosis factor alpha and interferon gamma and MODS, as indicated by a decrease in arterial oxygen pressure (lung) and an increase in plasma concentrations of bilirubin and alanine aminotransferase (liver), creatinine and urea (kidney), lipase (pancreas), and creatine kinase (heart or skeletal muscle) . There was also the expression of inducible nitric oxide synthase in these organs, circulatory failure, and 50% mortality . These effects were not observed after administration of PepG or LTA alone . Even a high dose of LTA (10 mg/kg) causes only circulatory failure but no MODS . Thus, our results demonstrate that the two bacterial wall components, PepG and LTA, work together to cause systemic inflammation and multiple systems failure associated with Gram-positive organisms.

Med J Aust, 1995 Oct 16, 163(8), 412 - 4
Changing epidemiology of methicillin-resistant Staphylococcus aureus in Western Australia; Riley TV et al.; OBJECTIVE: To assess the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) in Western Australia . DESIGN: Retrospective review of statutory notification data . SETTING: Western Australia (WA), 1993 . OUTCOME MEASURES: Notification rates, antibiotic resistance patterns and classification of isolates as imported or WA MRSA strains on the basis of antibiotic susceptibility . RESULTS: There were 204 notifications of MRSA, 78% of which were classified as WA MRSA . Three outbreaks of MRSA infection and colonisation occurred in separate WA hospitals . Notification rates per 100,000 were highest in the rural regions: the Kimberley (86.32), Goldfields (62.47), Mid West (37.21) and Pilbara (27.38) regions; and lowest in the metropolitan regions (5.52) . All MRSA isolates were susceptible to vancomycin . Most imported strains were susceptible to amikacin, bacitracin, chloramphenicol, framycetin, fusidic acid and novobiocin, but only 23% to gentamicin . WA MRSA strains remained predominantly susceptible to all antibiotics tested, except beta-lactams, erythromycin and tetracycline, but a few strains resistant to rifampicin (1%) and fusidic acid (3%) appeared in the second half of 1993 . CONCLUSIONS: The epidemiology of MRSA in WA is changing rapidly, with increases in both the numbers of notifications and the proportion from country regions . A new strain of MRSA (WA MRSA) that is less resistant to antibiotics than imported MRSA has emerged and is threatening the State's success in preventing establishment of MRSA in its hospitals.

J Neurosci Res, 1995 Oct 15, 42(3), 335 - 42
Increased production of gelatinase B (matrix metalloproteinase-9) and interleukin-6 by activated rat microglia in culture; Gottschall PE et al.; Activated macrophages produce several matrix metalloproteinases (MMPs), a family of extracellular matrix (ECM)-degrading enzymes, during wound healing and in other inflammatory states . In response to brain injury, brain microglia become "activated," in a way similar to peripheral tissue macrophages, a process which includes differentiation and probably invasion and proliferation . Little is known about the ECM-degrading MMPs that are secreted by microglia upon activation . Thus, it was of interest to determine whether activated microglia secrete MMPs . Conditioned media samples obtained from cultured microglia that were stimulated with various activating agents were subjected to gelatin-substrate zymography . Microglia constitutively express low levels of a 94-kDa gelatinase (GLase) activity . Treatment with LPS, zymosan, and fixed Staphylococcus aureus for 24 hr stimulated the activity of the 94-kDa GLase, 4-20-fold, in a dose-dependent manner . Addition of INF gamma inhibited the LPS-stimulated activity of MMP-9 . LPS, zymosan, and fixed Staphylococcus aureus also stimulated the secretion of IL-6 from microglia in a dose-dependent manner . The 94-kDa GLase activity was Ca++ dependent, it was inhibited by 1,10-phenanthroline, and it was activated by organomercurial compounds . When immunoblots were performed using specific antisera against the 94-kDa gelatinase B (MMP-9) with untreated and LPS-stimulated conditioned medium samples, a 94-kDa immunopositive band was observed . Thus, it appears that the 94-kDa GLase is gelatinase B (MMP-9) . These results indicate that activators of peripheral macrophages are potent secretagogues for the MMPs in cultured microglia . The ability of activated microglia to secrete MMPs suggests that these enzymes may play an important function in the brain parenchyma during inflammatory states.

FEMS Microbiol Lett, 1995 Oct 15, 132(3), 221 - 8
FemA of Staphylococcus aureus: isolation and immunodetection; Johnson S et al.; FemA, a cytoplasmic protein necessary for the expression of methicillin resistance in Staphylococcus aureus and also involved in the biosynthesis of staphylococcal cell walls, was detected and quantified in several S . aureus strains under different growth conditions by Western immunoblot . Two types of antigens were used for the production of polyclonal antibodies against FemA: (i) a synthetic peptide comprising 14 amino acids of its C-terminal sequence; and (ii) FemA isolated by preparative gel electrophoresis and electroelution from an overproducing staphylococcal strain . Immunodetection revealed that all investigated strains, either methicillin-resistant or susceptible, expressed FemA during the exponential growth phase in varying amounts . In the stationary phase, the FemA content was diminished . Strains in which femA was inactivated by insertion of Tn551 into the control region of the femAB operon still expressed about 10% of the protein compared to their parent strains . Tn551 insertion in the middle of the femB gene did not affect the FemA expression . In 40 methicillin-susceptible and 6 resistant clinical isolates of S . aureus, the FemA content or its affinity to the antibodies was reduced compared to laboratory parent strains . In susceptible strains, an additional protein of higher molecular weight, present in large quantities, was also able to bind the FemA antibodies . Such a protein was also present in methicillin-resistant isolates, although it was not as pronounced as in the susceptible strains.

Dtsch Med Wochenschr, 1995 Oct 13, 120(41), 1395 - 8
{Herpetic eczema in pregnancy}; Wollenberg A et al.; HISTORY AND FINDINGS: In a 32-year-old 26-week pregnant woman with atopic eczema densely packed blisters, ca . 1 mm in diameter, developed within the eczema areas, typical of herpetic eczema . This diagnosis was confirmed by identifying the virus under the electronmicroscope and demonstrating herpes simplex virus DNA in the polymerase chain reaction . TREATMENT AND COURSE: During intravenous aciclovir administration (5 mg/kg three times daily for 9 days) the blisters quickly healed . As a bacterial superinfection was suspected (later confirmed by culturing Staphylococcus aureus from a skin swab) the patient was also given erythromycin (500 mg four times daily by mouth for 7 days), as well as fusidic acid cream locally . She delivered a healthy child in the 37th week of pregnancy . CONCLUSION: Herpetic eczema is a serious complication of chronic eczema . If it occurs during pregnancy, the risk or fetal damage by aciclovir must be balanced against the risk of intrauterine herpes simplex infection.

Pharmacol Res, 1995 Oct, 32(4), 201 - 7
Effects of rebamipide on gastric cell damage by Helicobacter pylori-stimulated human neutrophils; Han BG et al.; Helicobacter pylori stimulated human neutrophils to produce oxygen radicals as evidenced by the production of chemiluminescence in the presence of luminol . The capacity of H . pylori to produce oxygen radicals from neutrophils was much higher than that of Escherichia coli and Staphylococcus aureus and is almost as strong as that of PMA . Rebamipide (2-(4-chlorobenzoylamino)-3-{2-(1H)-quinolinon-4-yl} propionic acid) suppressed the chemiluminescence produced by H . pylori-stimulated neutrophils and also suppressed the chemiluminescence produced by a cell-free xanthine/xanthine oxidase reaction with luminol . Thus, it is indicated that this drug has the action of scavenging oxygen radicals . Gastric mucosal cells labelled with a fluorescent dye were damaged by the incubation of the cells with neutrophils and H . pylori, and this damage was protected by rebamipide . The protection of cell damage was ascertained as a decrease in the release of fluorescent dye into the incubation medium and a reduction in the distortion of cell geometry . The data suggest that H . pylori induce human neutrophils to produce oxygen radicals which are responsible for gastric mucosal cell damage and that rebamipide removes the oxygen radicals produced from H . pylori-activated neutrophils and thus reduces the gastric mucosal cell damage . These effects may account for the ulcerogenesis action of H . pylori and for part of the mechanism of the anti-ulcer action of rebamipide.

Perit Dial Int, 1995 Oct-Dec, 15(8), 320 - 7
Phagocytosis and killing of suspended and adhered bacteria by peritoneal cells after dialysis; Calame W et al.; OBJECTIVE: To determine the effect of dialysis fluid containing various glucose concentrations on the phagocytosis and killing of Staphylococcus aureus by rat peritoneal cells under conditions mimicking the in vivo situation . DESIGN: Phagocytosis and killing were evaluated by quantitation of the killing capacity of macrophages after in vivo phagocytosis of the bacteria as well as by an in vitro flow cytometric assay of the phagocytosis and killing of adhered bacteria by peritoneal cells . ANIMALS: Male Wistar rats . MAIN OUTCOME MEASURE: It was expected that the intraperitoneal administration of dialysis fluid would impair the capacity of peritoneal cells to eliminate bacteria . RESULTS: The first test revealed no effects of glucose concentration or dwell time on the killing of phagocytosed bacteria by macrophages, median percentages ranging between 29% and 64% . In the second series of experiments no effect of glucose concentration on the phagocytosis and killing of adhered bacteria was found either; however, longer dwell times significantly enhanced both the phagocytosis (at a dwell time of 1 hour, under 20%; at dwell times of 4 or 18 hours, above 20%, p < 0.02) and the killing (at a dwell time of 1 hour, under 53%; at dwell times of 4 and 18 hours, above 70%, p < 0.01) . CONCLUSIONS: Glucose concentration has no effect on the phagocytosis and killing of Staphylococcus aureus, whereas the dwell time significantly enhances both of these functional capacities of peritoneal cells if the bacteria are adhered to surfaces.

Arq Gastroenterol, 1995 Oct-Dec, 32(4), 186 - 90
{Liver abscess in childhood: report of 8 cases}; Pereira RM et al.; The authors present eight cases of children with liver abscess diagnosed in the Pediatric Nursery of the "Hospital das Clinicas", State University of Campinas, SP, Brazil, during eight years . Five children were younger than five years . They were four boys and four girls . The symptomatology was inespecific . Staphylococcus aureus was the more frequent etiologic agent and in two cases it was seen Ascaris lumbricoides worm inside the abscess . The findings of ultrasound and computadorized tomography were similar . In most cases, were employed the association of broad spectrum antibiotics and percutaneous draining, and a good clinical outcome was observed in all patients.

Minerva Stomatol, 1995 Oct, 44(10), 445 - 7
Butyric acid, a metabolic end product of anaerobic bacteria, inhibits B-lymphocyte function; Eftimiadi C et al.; The effect on B-lymphocytes of butyric acid, a metabolic by-product formed by selected anaerobic bacteria of the subgingival plaque and present in mM amounts in the crevicular fluid of patients affected by adult periodontitis was investigated . A dose-dependent inhibition of both mitogenesis and Ig production in B-cells challenged with formalinized Staphylococcus aureus, strain Cowan I (SAC), was observed . A role of this molecule in the downregulation of the local immune response in periodontal patients is proposed.

Can Vet J, 1995 Oct, 36(10), 619 - 23
Detection of Staphylococcus aureus in bulk tank milk using modified Baird-Parker culture media; Ollis GW et al.; The purpose of this project was to evaluate the use of 2 selective/differential culture media for detecting Staphylococcus aureus in bulk tank milk . One medium was Baird-Parker agar base supplemented with egg york tellurite emulsion and acriflavine . The other medium was Baird-Parker agar base supplemented with rabbit plasma/bovine fibrinogen and acriflavine . An increased inoculum of bulk tank milk (0.3 mL) was used to enhance the detection of S . aureus in samples containing low numbers of organisms . The sensitivity and specificity for detecting S . aureus in bulk tank milk were 94.8% and 100%, respectively, using Baird-Parker agar base supplemented with egg yolk tellurite emulsion and acriflavine, and 89.7% and 100%, respectively, using Baird-Parker agar base supplemented with rabbit plasma/bovine fibrinogen and acriflavine . Both media are practical for detecting S . aureus in bulk tank milk and monitoring its spread in lactating dairy herds in Alberta.

Antimicrob Agents Chemother, 1995 Oct, 39(10), 2289 - 94
Vancomycin or vancomycin plus netilmicin for methicillin- and gentamicin-resistant Staphylococcus aureus aortic valve experimental endocarditis; Perdikaris G et al.; Using a rabbit model of aortic valve endocarditis, we studied the efficacy of vancomycin alone or in combination with netilmicin and/or rifampin against a methicillin- and gentamicin-resistant strain of Staphylococcus aureus (MGRSA) . Antibiotics were given for 6 to 12 days, as follows: vancomycin (15 mg/kg of body weight every 12 h {BID} intravenously), vancomycin plus netilmicin (2.5 mg/kg BID intramuscularly), vancomycin plus rifampin (10 mg/kg BID intramuscularly), and vancomycin plus netilmicin plus rifampin at the same routes, dosages, and schedules mentioned above . Netilmicin was given to two additional groups at a higher dosage (6 mg/kg every 24 h intramuscularly) alone or in combination with vancomycin (15 mg/kg BID intravenously) for 12 days . All regimens resulted in undetectable bacterial counts in a significant proportion of vegetations (except netilmicin alone) or reduced the bacterial counts in the vegetations compared with the counts in the untreated controls (P<0.01 to P<0.001) . No resistance to rifampin or netilmicin developed during therapy . It is concluded that in the treatment of experimental aortic valve endocarditis caused by MGRSA (i) vancomycin as monotherapy is as efficacious as the triple combination, (ii) the addition of netilmicin (once daily or BID) to vancomycin does not improve the efficacy of the latter antibiotic, even in the presence of rifampin, and (iii) a 12-day course in more effective than a 6-day one, but not at a statistically significant level.

Biochem Mol Biol Int, 1995 Oct, 37(3), 527 - 35
Structural and functional features of noxiustoxin: a K+ channel blocker; Gurrola GB et al.; Binding of {125I}-Noxiustoxin to rat brain synaptosome membranes and displacement with synthetic peptides corresponding to the amino acid sequence of Noxiustoxin, show that the N-terminal segment of this toxin is implicated in the recognition of brain K(+)-channels . Cleavage of NTX with cyanogen bromide, endopeptidase V8 from Staphylococcus aureus and endopeptidase Lys-C support these findings . On the contrary, a synthetic C-terminal tetradecapeptide of charybodotoxin (ChTX), shows that in this K(+)-channel blocker, the C-terminal region, rather than the N-terminal is capable of displacing 125{I}-NTX binding to brain membranes.

J Vet Pharmacol Ther, 1995 Oct, 18(5), 340 - 5
Tilmicosin antibacterial activity and pharmacokinetics in cows; Ziv G et al.; The minimal inhibitory concentration (MIC) of tilmicosin for 90% of 112 Staphylococcus aureus isolates from the bovine udder was 0.78 microgram/mL and 149 of 164 (90.8%) other gram-positive udder pathogens were inhibited by tilmicosin concentrations < 3.12 micrograms/mL . The MIC of the drug for 19 of 22 S . aureus isolates was < 0.78 microgram/mL when the test was conducted using Mueller-Hinton (MH) agar or MH agar containing 7.5% skimmed milk . Acute cardiac toxicity followed intravenous (i.v.) injection of the drug at 10 mg/kg to 3 cows, but animals appeared clinically normal within 30 min after treatment . The pharmacokinetics of i.v.-administered tilmicosin is typical for the macrolide class of antibiotics, i.e . low serum drug concentrations and a large volume of distribution (> 2.0 L/kg) . The elimination half-life (t1/2 beta) values for 3 cows were 46.4, 56.0 and 72.8 min . The drug was administered subcutaneously (s.c.) to 5 cows at 10 mg/kg; the elimination half-life (t1/2el) was 4.18 +/- 0.55 h and the mean s.c . bioavailability was 22% . Rapid and extensive penetration of tilmicosin from blood into milk, and slow elimination from the milk were among the characteristic kinetic features of the drug after i.v . and s.c . administration . Tilmicosin was injected s.c . at 10 mg/kg once to 9 cows after the last milking of lactation; dry udder secretion samples were collected daily for 11 consecutive days and assayed microbiologically . Concentrations of drug > 0.78 microgram/mL were found in the secretion for 8-9 days after dosing . Systemic side-effects were not observed after s.c . drug administration.

Am J Infect Control, 1995 Oct, 23(5), 306 - 9
Mupirocin ointment with and without chlorhexidine baths in the eradication of Staphylococcus aureus nasal carriage in nursing home residents; Watanakunakorn C et al.; BACKGROUND: Mupirocin ointment has been shown to be effective in eradicating Staphylococcus aureus nasal carriage in residents of a long-term care facility . Antiseptic soaps have been used as adjunct to this therapy . We compared the efficacy of short-term intranasal mupirocin ointment with and without chlorhexidine baths in the eradication of S . aureus nasal carriage with follow-up for 12 weeks . METHODS: Residents in four nursing homes known to have endemic methicillin-resistant S . aureus were screened for nasal carriage of S . aureus . Residents who had anterior nares cultures positive for S . aureus on two separate occasions were divided into two groups . Both groups received intranasal mupirocin ointment twice daily for 5 days and one group also received chlorhexidine baths for the first 3 days . Cultures of anterior nares, axilla, and groins were performed before treatment and 1 day and 1, 4, 8, and 12 weeks after treatment . RESULTS: After treatment, S . aureus nasal carriage was eradicated in all residents . Recolonization with S . aureus had occurred at 12 weeks in 24% of residents receiving mupirocin ointment alone (6/25) and in 15% of residents receiving mupirocin ointment plus chlorhexidine baths (4/27) . CONCLUSIONS: A short course of mupirocin ointment was effective in eradicating nasal carriage of S . aureus in nursing home residents . There were no statistical differences in efficacy between the two regimens with respect to the eradication of nasal carriage and prevention of recolonization with S . aureus.

Pediatr Nephrol, 1995 Oct, 9(5), 647 - 62
Host defences in continuous ambulatory peritoneal dialysis and the genesis of peritonitis; Cameron JS; Continuous ambulatory peritoneal dialysis (CAPD) has come to be extensively used for the treatment of end-stage renal failure in children, and especially infants, such that now more than half of children on dialysis worldwide receive treatment by this means . Peritonitis, however, is commoner in children than in adults receiving treatment, and is a major source of morbidity and treatment failure in children started on CAPD . Only recently has the immunology of the normal peritoneum been studied extensively, with the need to assess the impact of the installation of large volumes of fluid into the peritoneal sac during dialysis . The main phagocytic defences of the peritoneum depend upon a unique set of macrophages which are present free in the peritoneal fluid but also in the submesothelium and in perivascular collections together with B lymphocytes in the submesothelial area . Both the number of macrophages per unit volume and the concentration of opsonic proteins, such as IgG, complement and fibronectin, are reduced to between only 1% and 5% when dialysis fluid is continuously present in the peritoneal sac . In addition, the fluids used for CAPD are toxic to both macrophages and to mesothelial cells . Thus minor degrees of contamination frequently lead to peritonitis and in addition the majority of patients have catheters inserted in their peritoneum which become colonised with organisms capable of producing exopolysaccharide (slime), which promotes adhesion of the organism to the plastic and protects them against phagocytic attack and the penetration of antibiotics . Thus the peritoneum is in a state of continual inflammation, as well as being a markedly more vulnerable site than the normal peritoneum to the entry of organisms . Whether clinical peritonitis appears in this state of chronic contamination probably depends on perturbation in the balance between host defences and the organism . Whilst Staphylococcus epidermidis is the commonest cause of peritonitis, Staphylococcus aureus and Gram-negative organisms are much more serious and more frequently lead either to temporary catheter removal or discontinuation of dialysis altogether . This review describes the peritoneal defences in relation to the genesis of peritonitis.

Pediatr Nephrol, 1995 Oct, 9(5), 609 - 11
Effect of rifampin on Staphylococcus aureus colonization in children on chronic peritoneal dialysis; Hanevold CD et al.; The efficacy of rifampin in eliminating Staphylococcus aureus colonization was evaluated in a pediatric peritoneal dialysis population . Six children with documented nasal colonization were treated for 7 days with rifampin and cloxacillin . Although antimicrobial therapy eliminated nasal carriage in all patients, recolonization occurred in 66% . Exit site colonization proved difficult to eradicate with negative cultures documented in only 3 of 5 children after rifampin/cloxacillin therapy . Although S . aureus carriage is a risk factor for S . aureus infections, efforts to eradicate carriage with rifampin are hindered by rapid recolonization.

J Clin Microbiol, 1995 Oct, 33(10), 2723 - 7
Comparison of MIDI Sherlock system and pulsed-field gel electrophoresis in characterizing strains of methicillin-resistant Staphylococcus aureus from a recent hospital outbreak; Leonard RB et al.; An outbreak of methicillin-resistant Staphylococcus aureus infections at the University of Utah Health Sciences Center occurred over a 7-month period . While the isolates phenotypically appeared to be similar in gross morphology and have similar Vitek antibiotic susceptibility patterns, two additional methods of strain characterization were evaluated to enhance the epidemiological investigation: pulsed-field gel electrophoresis and gas chromatography with the MIDI Sherlock system . Sherlock uses gas chromatography to qualitatively and quantitatively analyze the cellular fatty acid composition of organisms and creates two-dimensional plots based on principal-component analysis to define groups of closely related organisms . All isolates were also evaluated by digesting their chromosomal DNAs with the low-frequency-cutting enzyme SmaI and separating the restriction fragments by contour-clamped homogeneous electric field gel electrophoresis . Sample preparation for this pulsed-field gel electrophoresis included a novel cell lysis procedure involving achromopeptidase, greatly reducing the turnaround time . Isolates tested were recovered from the following: 45 suspected outbreak patients, 6 hospitalized patients believed to be unrelated to the outbreak, 6 patients from outside the hospital, and one health care practitioner implicated in the outbreak . Of 45 phenotypically similar suspect strains, 43 clustered tightly on the Sherlock two-dimensional plot . All outbreak patient isolates were also identical by pulsed-field gel electrophoresis with the exception of the same two outliers identified by Sherlock . In this epidemiologic investigation, we found an excellent correlation between the Sherlock and pulsed-field gel electrophoresis results for strain characterization of methicillin-resistant S . aureus.

Int Immunol, 1995 Oct, 7(10), 1691 - 8
ICAM-1 is required for T cell proliferation but not for anergy or apoptosis induced by Staphylococcus aureus enterotoxin B in vivo; Gonzalo JA et al.; The response of T lymphocytes to superantigens requires expression of the appropriate TCR V beta gene products as well as the establishment of cellular interactions mediated by adhesion molecules . To study the role of intercellular adhesion molecule (ICAM)-1 in the response in vivo to superantigens, we have analyzed the effects induced by the bacterial superantigen Staphylococcus aureus enterotoxin B (SEB) in mice which have been made genetically deficient in ICAM-1 . SEB treatment of wild-type mice causes proliferation, deletion and anergy of the SEB-reactive V beta 8+ T cell population . Here we show that cellular interactions mediated by ICAM-1 are not essential for the induction of anergy or for the deletion of CD4+ V beta 8+ or CD8+ V beta 8+ T cells, but are required for the proliferation of these peripheral T lymphocytes . This is the first demonstration in vivo that the absence of the co-stimulatory signals provided by the interaction of ICAM-1 with its specific ligands impairs the proliferation of SEB-reactive T cells . Interestingly, our study showed that SEB-induced proliferation of CD8+ V beta 8+ T cells from lymph nodes (not from spleen) is independent of the interactions mediated by ICAM-1.

J Membr Biol, 1995 Oct, 147(3), 233 - 9
Low conductance states of a single ion channel are not 'closed'; Korchev YE et al.; We have used a polymer-exclusion method to estimate the sizes of the high- and low-conductance states of Staphylococcus aureus alpha-toxin channels across planar lipid bilayers . Despite a > 10-fold difference in conductance between high- and low-conductance states, the size differs by < 2-fold . We conclude that factors other than the dimensions have a strong influence on the conductance of alpha-toxin channels . We also show that the high conductance state is destabilized by the presence of high molecular weight polymers outside the channel, compatible with the removal of channel water as the high conductance state "shrinks" to the low conductance state.

Eur J Surg, 1995 Oct, 161(10), 721 - 3
Infective complications after minor operations in patients infected with HIV: role of CD4 lymphocytes in prognosis; Emparan C et al.; OBJECTIVE: To find out the incidence of wound infection in patients with HIV and reduced counts of CD4 lymphocytes . DESIGN: Open study . SETTING: University hospital, Spain . SUBJECTS: 70 patients with HIV infection and enlarged lymph nodes . INTERVENTIONS: Biopsy of lymph nodes and withdrawal of a sample of blood for counts of CD4 lymphocytes and neutrophils . MAIN OUTCOME MEASURE: Development of infection at the biopsy site, and correlation of infecting organism with culture taken at the time of biopsy . RESULTS: Patients were divided into three groups depending on their CD4 count: more than 500 cells/ml (n = 26), 200-500 cells/ml (n = 24), and less than 200 cells/ml (n = 20) . Their neutrophil counts were 5.1, 3.8, and 2.5 x 10(9)/1, respectively . There were found four wound infections (6%); 2 were in the group with more than 500 CD4 cells/ml, and these were caused by Staphylococcus aureus (which had been grown from nodes in 6 patients at the time of biopsy) . The other 2 were in the group with less than 500 cells/ml and these were caused by Mycobacterium tuberculosis; cultures of the nodes had shown Staphylococcus epidermidis (n = 3) and M tuberculosis (n = 17) . There were no infections in the group with 200-500 CD4 cells/ml, in which S epidermidis (n = 5) and M tuberculosis (n = 8) had been cultured from the lymph nodes . CONCLUSIONS: The CD4 count was of no prognostic importance in the development of wound infection, but severe depression of the CD4 count may increase the risk of atypical wound infections.

Nippon Seikeigeka Gakkai Zasshi, 1995 Oct, 69(10), 1004 - 13
{Development and progression of pyogenic spondylitis in a canine experimental model}; Koh S; An experimental model was prepared to investigate the process of inflammation in pyogenic spondylitis . Forty-seven mongrel dogs were used, involving 24 mature and 23 immature dogs . Under intravenous pentobarbital anaesthesia, the lumbar vertebral bodies were approached posterolaterally and inoculated using a small piece of gauze soaked in a staphylococcus aureus suspension . Roentgenographic and histological examinations were regularly performed for 24 weeks after the inoculation . Histologically, acute inflammation started within 1 or 2 weeks, and subsided by 5 or 6 weeks in both the mature and immature dogs . In 55% of the dogs, the inflammation was confined within the vertebral body, in 10% it invaded into the intervertebral disc, and in 35% inflammation invaded into the anterior longitudinal ligament . In the immature dogs, thickening of the trabeculae and the anterior cortex was observed around the inflammatory focus more often than in the mature dogs . The epiphyseal line acted as a barrier against invasion by the inflammation in the immature dogs . However, direct invasion of the inflammatory process into the disc could have occurred through the vascular buds which were the terminal branches of the metaphyseal artery close to the disc in both the mature and immature dogs . In contrast to the results reported by Ohno who inoculated the lumbar discs of mongrel dogs with staphylococcus aureus, in the present study, the disc space remained intact and was replaced by fibrous tissue . Consequently, it was concluded that pyogenic spondylitis should be defined as a different clinical entity from discitis.

J Hosp Infect, 1995 Oct, 31(2), 135 - 41
Labile antibiotic resistance in Staphylococcus aureus; Noble WC et al.; Twenty-two isolates of Staphylococcus aureus, recovered from patients over a period of about one year, exhibited low level resistance to mupirocin, developed a salmon pink colour on culture and 21 had the same phage-type . However the plasmid profiles and associated antibiotic resistances differed . Digestion of cellular DNA with SmaI showed that two isolates from a single patient had a markedly different pattern to the remainder, and that six others differed by one band, though these formed groups of one and five isolates . This episode apparently represents a small outbreak of colonization or infection, which would have been missed but for the unusual pigmentation of the isolates recovered, and illustrates the difficulties of relying on a single typing system.

J Hosp Infect, 1995 Oct, 31(2), 123 - 34
Epidemiological analysis of strains of methicillin-resistant Staphylococcus aureus (MRSA) infection in the nursery; prognosis of MRSA carrier infants; Mitsuda T et al.; Forty-five neonates who carried methicillin-resistant Staphylococcus aureus (MRSA) were studied . Retrospective molecular analysis using pulsed-field gel electrophoresis showed three separate MRSA epidemics in the nursery . Strains of MRSA isolated from the neonates were also isolated from the hospital environment and health care providers . Clinical manifestations included skin pustules (eight patients), conjunctivitis (four patients), or other minor infections (two patients) . No neonate developed systemic infection . The prevalence of MRSA decreased with age . At one year, three (14.3%) of 21 infants that had carried MRSA at six days remained carriers and only two (1.1%) of 180 infants in a control 'S . aureus-negative at six days' group carried MRSA.

Nihon Kyobu Shikkan Gakkai Zasshi, 1995 Oct, 33(10), 1058 - 63
{Microorganisms cultured from sputum and blood in association with episodes of fever during anti-cancer therapy in patients with lung cancer}; Rikimaru T et al.; We examined the clinical features and significance of pathogenic microbes isolated from sputum and blood of patients with lung cancer during anti-cancer therapy . Pathogenic bacteria were more likely to be isolated from patients with episodes of fever than from afebrile patients . The major species of bacteria isolated from sputum were Staphylococcus aureus, including methicillin-resistant strains, and Gram-negative bacilli, which are known to be frequently involved in hospital-acquired infections . The presence of an indwelling central venous catheter for intravenous hyperalimentation was an important risk factor for the development of a febrile episode, which indicates that bacteremia was a major cause of fever . In one quarter of the blood cultures from the patients with persistent fever, various species of pathogenic microbes were recovered, one-third of which were fungi . Bacteriological examinations done before and after the introduction of granulocyte-colony stimulating factor (G-CSF) revealed that strains of Klebsiella spp . decreased and those of methicillin-resistant S . aureus increased . There was no firm evidence that G-CSF decreased the incidence of episodes of fever . However, remains G-CSF may a allow the dose intensity of anti-cancer agents to be increased, which would lead to severe leukocytopenia . However, more detailed investigation is needed to clarify the role of G-CSF against bacterial infection during anti-cancer therapy.

Indian J Med Res, 1995 Oct, 102, 156 - 8
Rapid identification of colonization factor antigens I & II of enterotoxigenic Escherichia coli by coagglutination test; Ram S et al.; Specific antisera for colonization factor antigens (CFA/I and CFA/II) were adsorbed to Staphylococcus aureus Cowan I strain, ATCC 12598 to make coagglutination (CoA) reagents for detection of CFAs in enterotoxigenic Esch . coli (ETEC) isolates . Among 1782 strains of Esch . coli isolated from patients with acute diarrhoea, 238 (13.4%) strains exhibited CFA expression . Most prevalent CFA/I positive serogroups were 015, 0148, 0153, 020, 0128, 0114 and 078 . CFA/II was detected among isolates of serogroup 080, 085, 06 and 08 . Ten ETEC isolates each of serogroups 04, 07, 061, 068, 0117 and 0158 did not show presence of CFA/I or CFA/II . CoA technique proved an appropriate, rapid diagnostic tool which can be used for screening large number of Esch . coli isolates in epidemiological studies.

Protein Expr Purif, 1995 Oct, 6(5), 671 - 8
High-yield expression, refolding, and purification of penicillin-binding protein 2a from methicillin-resistant Staphylococcus aureus strain 27R; Frank LJ et al.; The mecA-27R gene, which encodes PBP2a from methicillin-resistant Staphylococcus aureus strain 27R, was modified to remove the putative N-terminal membrane-spanning region, cloned into the T7 RNA polymerase expression vector pET11d, and used to transform Escherichia coli strain BL21(DE3) . The majority of PBP2a was expressed in the form of inclusion bodies, which were extracted, denatured, and refolded . The protein was then purified by anion-exchange and size-exclusion chromatography . A 6-liter culture of induced E . coli provided 37 mg of purified PBP2a which was greater than 99% pure . Binding affinities for {3H}benzylpenicillin, imipenem, and L-695,256 (a beta-lactam with high affinity for PBP2a) were shown to be comparable to PBP2a found in membrane preparations of S . aureus strain 27R . A direct binding assay, using 14C-labeled L-695,256 was developed and used to show stoichiometric binding to the refolded, soluble PBP2a . In addition, electrospray mass spectrometry showed that 100% of the refolded PBP2a was covalently bound to the beta-lactam in a stoichiometric fashion . Finally, two mutations of the putative active-site serine showed the predicted loss of covalent binding of the beta-lactam to the PBP2a, demonstrating the high specificity of the soluble binding assay.

Biosci Biotechnol Biochem, 1995 Oct, 59(10), 1981 - 2
Identification of the positions of disulfide bonds of chitinase from a marine bacterium, Alteromonas sp . strain O-7; Hayashi K et al.; Extracellular chitinase from marine Alteromonas sp . strain O-7 is unique because of the activation by four major cations contained in sea water, such as Na+, K+, Mg2+, and Ca2+ . The positions of S-S bonds of Alteromonas chitinase were identified . Alteromonas chitinase was fragmented by TPCK-trypsin and Staphylococcus aureus V8 protease . The amino acid and sequence analyses of three peptides showed that the positions of disulfide bonds are Cys(94)-Cys(99), Cys(174)-Cys(196), and Cys(386)-Cys(395).

Rinsho Byori, 1995 Oct, 43(10), 1061 - 5
{Methicillin-resistant Staphylococcus aureus forming the fried egg appearance colonies isolated from a patient with septicemia}; Tanimoto A et al.; Slower growing and peculiar colonies, "fried egg" appearance, of different sizes were temporally grown on a sheep blood agar plate from a clinical blood sample obtained from a patient with mediastinitis due to infection by methicillin-resistant Staphylococcus aureus (MRSA) following the graft replacement of thoracic aorta . The organisms revealed morphologically characteristic "fried egg" appearance, mainly Gram-positive cocci including some Gram-negative cells and cell-debris, catalase test positive and coagulase test positive as well as having the identical profiles for S . aureus on biochemical examination with VITEK Systems . The rough surfaced cell walls with unequal thickness of the organisms were observed in an electronmicroscope . According to these bacteriological studies, the organisms were identified as a mutant of S . aureus which had many similarities to staphylococcal L-form . Being resistant to oxacillin, instead of methicillin susceptibility test, and possessing a mecA gene, the organisms were confirmed to belong to MRSA . It is likely that this mutant of S . aureus was induced by a long term medication of sulfamethoxazole-trimetoprim, since the patient had been receiving the drug during 5 weeks when the colonies were first detected.

Blood, 1995 Oct 1, 86(7), 2509 - 15
Results of a phase I/II trial of recombinant human granulocyte-macrophage colony-stimulating factor in very low birthweight neonates: significant induction of circulatory neutrophils, monocytes, platelets, and bone marrow neutrophils; Cairo MS et al.; Neonates, especially those of very low birthweight (VLBW), have an increased risk of nosocomial infections secondary to deficiencies in development . We previously demonstrated that granulocyte-macrophage colony-stimulating factor (GM-CSF) production and mRNA expression from stimulated neonatal mononuclear cells are significantly less than that from adult cells . Recombinant murine GM-CSF administration to neonatal rats has resulted in neutrophilia, increased neutrophil production, and increased survival of pups during experimental Staphylococcus aureus sepsis . In the present study, we sought to determine the safety and biologic response of recombinant human (rhu) GM-CSF in VLBW neonates . Twenty VLBW neonates (500 to 1,500 g), aged < 72 hours, were randomized to receive either placebo (n = 5) or rhuGM-CSF at 5.0 micrograms/kg once per day (n = 5), 5.0 micrograms/kg twice per day (n = 5), or 10 micrograms/kg once per day (n = 5) given via 2-hour intravenous infusion for 7 days . Complete blood counts, differential, and platelet counts were obtained, and tibial bone marrow aspirate was performed on day 8 . Neutrophil C3bi receptor expression was measured at 0 and 24 hours . GM-CSF levels were measured by a sandwich enzyme-linked immunosorbent assay at 2, 4, 6, 12, and 24 hours after the first dose of rhuGM-CSF . At all doses, rhuGM-CSF was well tolerated, and there was no evidence of grade III or IV toxicity . Within 48 hours of administration, there was a significant increase in the circulating absolute neutrophil count (ANC) at 5.0 micrograms/kg twice per day and 10.0 micrograms/kg once per day, which continued for at least 24 hours after discontinuation of rhuGM-CSF . When the ANC was normalized for each patient's first ANC, there was a significant increase in the ANC on days 6 and 7 at each dose level . By day 7, all tested doses of rhuGM-CSF resulted in an increase in the absolute monocyte count (AMC) compared with placebo-treated neonates . In those receiving rhuGM-CSF 5.0 micrograms/kg twice per day, there was additionally a significant increase in the day 7 and 8 platelet count . Tibial bone marrow aspirates demonstrated a significant increase in the bone marrow neutrophil storage pool (BM NSP) at 5.0 micrograms/kg twice per day and 10.0 micrograms/kg once per day . Neutrophil C3bi receptor expression was significantly increased 24 hours after the first dose of rhuGM-CSF at 5.0 micrograms/kg once per day . The elimination half-life (T1/2) of rhuGM-CSF was 1.4 +/- 0.8 to 3.9 +/- 2.8 hours.(ABSTRACT TRUNCATED AT 400 WORDS)

Eur J Haematol, 1995 Oct, 55(4), 267 - 71
Methicillin-resistant Staphylococcus aureus: colonization and development of infection in patients with haematological disorders; Hagiwara S et al.; A retrospective study of 53 patients with haematological disorders whose bacterial cultures were positive for methicillin-resistant Staphylococcus aureus (MRSA), was performed to analyse the risk factors for MRSA infection, and the prognostic factors . Sixteen patients showed colonization by MRSA but never developed infection(C), 16 showed colonization and subsequent infection(C-I), while 21 had MRSA infection at the time of first culture (I) . Poor performance status, thrombocytopenia, increased serum urea nitrogen and decreased serum cholinesterase were more prominent in (I) than (C) + (C-I) . The risk factors associated with the development of infection from colonization were age and serum cholinesterase . In addition, lower respiratory tract infection as a type of infection, non-remission status of the haematological malignancy and an inappropriate antibiotic therapy were associated with a poor prognosis for MRSA infection.

Eur J Immunol, 1995 Oct, 25(10), 2825 - 9
CD27/CD70 interaction directly drives B cell IgG and IgM synthesis; Agematsu K et al.; CD27 is a T cell activation antigen expressed on a majority of peripheral blood T cells . CD27 is also expressed on a subpopulation of human B cells, and it is reported that CD27+ B cells secrete both IgG and IgM . CD70, a ligand for CD27, is expressed on activated T and B cells, suggesting an interaction between T and B cells via CD27/CD70 ligation . Here, we analyze B cell immunoglobulin synthesis using a CD70 transfectant and present functional data showing that B cells secrete large amounts of IgG and IgM as a result of the CD27/CD70 interaction . A flow cytometric analysis showed that CD27 expression was increased and CD70 was expressed on tonsillar and peripheral blood B cells after activation with Staphylococcus aureus Cowan strain (SAC) plus interleukin (IL-2) . In addition, the proliferation of B cells was enhanced mildly by the addition of CD70 transfectant, and its proliferation was blocked by anti-CD70 mAb . More importantly, the CD70 transfectant enhanced IgG and IgM production by purified B cells greatly in the presence of SAC plus IL-2 . The enhancement was completely blocked by the addition of either anti-CD70 mAb or anti-CD27 mAb . Strongly suggesting that the interaction of CD27 with its ligand, CD70, on B cells plays an important role in B cell growth and differentiation to produce IgG and IgM.

Eur J Biochem, 1995 Oct 1, 233(1), 179 - 85
The amino acid sequence and interaction with the nucleosome core DNA of transition protein 4 from boar late spermatid nuclei; Akama K et al.; The primary structure of transition protein 4 (TP4) from boar late spermatid nuclei was determined by automated Edman degradation of S-pyridylethylated protein and of peptides generated by cleavage with Staphylococcus aureus V8 protease, lysyl endopeptidase and CNBr . Boar TP4 is a basic protein consisting of a highly basic amino-terminal half (residues 1-73) and a less basic carboxy-terminal half (residues 74-138) . The latter half includes a highly hydrophobic segment, a four-times tandemly repeated sequence, N(G)QNKR(K)X, and a carboxy-terminal segment containing Trp126 . Ultraviolet absorption and CD spectra of TP4-rat-liver-nucleosome-core-DNA (double-stranded DNA) complexes suggest a TP4-induced local melting of DNA . Although at 1 mM NaCl TP4 brought about a slight stabilization of the DNA against thermal melting, a destabilization of the DNA was observed at 50 mM NaCl . From the results of quenching of tryptophan (Trp126) fluorescence of TP4 upon its binding to double-stranded and single-stranded boar liver nucleosome-core DNA at 50 mM NaCl, the apparent association constants for the binding of TP4 to double-stranded and single-stranded DNA were calculated to be 7.3 x 10(3) M-1 and 4.1 x 10(3) M-1, respectively . These results suggest that TP4, having different domain structures from TP1-3 and a higher affinity for double-stranded DNA, induces a local destabilization of DNA probably through the stacking of Trp126 with nucleic acid bases.

Lett Appl Microbiol, 1995 Oct, 21(4), 219 - 22
Antibacterial activity of flavanostilbenes against methicillin-resistant Staphylococcus aureus; Sato M et al.; Three phytochemical compounds (alopecurone A-C), flavanostilbenes which are produced by condensation between a hydroxyflavanone and a hydroxystilbene, were isolated as major components from the root of Sophora alopecuroides . They uniformly inhibited the growth of 21 strains of methicillin-resistant Staphylococcus aureus with minimum inhibitory concentrations of 3.13-6.25 micrograms ml-1.

Ann Surg, 1995 Oct, 222(4), 482 - 90; discussion 490-2
Major injury leads to predominance of the T helper-2 lymphocyte phenotype and diminished interleukin-12 production associated with decreased resistance to infection; O'Sullivan ST et al.; OBJECTIVE: Patients with serious traumatic injury and major burns and an animal model of burn injury were studied to determine the effect of injury on the production of cytokines typical of the T helper-2 lymphocyte phenotype as opposed to the T helper-1 phenotype and on the production of interleukin-12 . SUMMARY BACKGROUND DATA: Perturbations of natural and adoptive immunity are related to the increased susceptibility to infection manifested by seriously injured and burn patients . Earlier work has shown that impaired adoptive immunity after injury is characterized by diminished production of interleukin-2 (IL-2), a product of Th lymphocytes . Exposure of naive Th cells to certain antigens and cytokines causes conversion to either the Th-1 or the Th-2 phenotype . Th-1 cells produce IL-2 and interferon-gamma (IFN-tau) and initiate cellular immunity . Th-2 cells secrete interleukin-4 (IL-4) and interleukin-10 (IL-10) and stimulate production of certain antibodies . Conversion to the Th-1 phenotype is facilitated by IL-12, and conversion to the Th-2 phenotype is promoted by IL-4 . The authors believed that serious injury might cause conversion of Th cells to the Th-2 as opposed to the Th-1 phenotype rather than generalized Th suppression . METHODS: The authors studied circulating peripheral blood mononuclear cells (PBMC) from 16 major burn and 8 trauma patients on 32 occasions early after injury and from 13 age- and sex-matched healthy individuals for cytokine production after phytohemagglutinin stimulation . Also studied was a mouse model of 20% burn injury known to mimic the immune abnormalities seen in humans with burns . Splenocytes from burn mice, 10 to 12 per group, were studied after activation by concanavalin A or by the bacterial antigen Staphylococcus aureus Cowan strain I for cytokine production and cytokine messenger RNA expression as determined by reverse transcriptase polymerase chain reaction . Burn mice were compared with sham-burn controls and attention was focused on day 10 after burn injury, a time when IL-2 production and resistance to infection are highly suppressed . Finally, burn and sham-burn animals, 20 per group, were treated in vivo with IL-12 (25 ng daily for 5 days) and observed for mortality after septic challenge (cecal ligation and puncture {CLP}) performed on day 10 after injury . RESULTS: Peripheral blood mononuclear cells from burn and trauma patients produced less IFN-tau, the index cytokine of Th-1 cells, than PBMCs from healthy individuals 1 to 14 days after burn injury (SE = 77.6 +/- 16 pg/mL patients vs . 141.3 +/- 35 pg/mL controls, p < 0.05) . However, production of IL-4, the index cytokine of Th-2 cells, by patient PBMCs was increased (51.0 +/- 13.0 pg/mL patients vs . 26.9 +/- 2.5 controls, p < 0.05) . Splenocytes from mice 10 days after burn injury, when compared with sham-burn controls, showed diminished production of IL-2 (1.04 +/- 0.91 units/mL burns vs . 5.8 +/- 0.55 units/mL controls, p < 0.05) and IFN-tau (1.05 +/- 0.7 units/mL burns vs . 12.0 +/- 8.9 units/mL controls, p < 0.05) . However, burn splenocytes produced more IL-4 (2492 +/- 157.0 pg/mL burns vs . 672.0 +/- 22.7 pg/mL controls, p < 0.01) and IL-10 (695.2 +/- 20.8 pg/mL burns vs . 567.0 +/- 16.7 pg/mL controls, p < 0.05) . Splenocyte production of IL-12 was also reduced after burn (0.20 +/- 0.035 units/mL) as compared with sham burn (0.46 +/- 0.08 units/mL, p < 0.05) . The reduction in IL-2, IFN-tau, and IL-12 production by burn splenocytes was reflected by a tenfold decrease in expression of their respective cytokine mRNAs . In vivo IL-12 treatment of burn animals decreased mortality from CLP on day 10 after injury from 85% to 15% (sham-burn mortality after CLP, 15%, p < 0.05) and increased splenocyte IFN-tau production to supranormal levels . (ABSTRACT TRUNCATED)

J Infect Dis, 1995 Oct, 172(4), 1112 - 4
Comparison of cotton and cotton/rayon tampons for effect on production of toxic shock syndrome toxin; Schlievert PM; Studies were done to compare tampons made solely of cotton and made of both cotton and rayon for effect on growth of Staphylococcus aureus and production of toxic shock syndrome toxin-1 (TSST-1) . Under stationary in vitro conditions in which tampons were either oversaturated or 50% saturated with culture media, the same amount of or more TSST-1 was made with cotton tampons than with cotton/rayon tampons . Similarly, when tested with the tampon sac method, cotton tampons yielded the same amount of or more toxin than did the cotton/rayon tampons . Bacterial cell numbers generally paralleled toxin production . These data indicate that cotton tampons neither prevent TSST-1 production nor significantly adsorb toxin onto the fibers to make toxin unavailable to cause toxic shock syndrome, in contrast to results of a previous study.

J Invest Dermatol, 1995 Oct, 105(4), 536 - 42
Staphylococcal enterotoxin B upregulates expression of ICAM-1 molecules on IFN-gamma-treated keratinocytes and keratinocyte cell lines; Wakita H et al.; The effects of staphylococcal enterotoxin B (SEB), a Staphylococcus aureus-derived bacterial superantigen, on expression of intercellular adhesion molecule-1 (ICAM-1) were examined in cultured normal and transformed (DJM-1 cells) human keratinocytes by flow cytometry, confocal microscopy, digital image processing, and reverse transcriptase-polymerase chain reaction . SEB significantly upregulated ICAM-1 expression in the interferon-gamma (IFN-gamma)-pretreated, HLA-DR-positive normal keratinocytes and DJM-1 cells in a dose-dependent manner, but not in the untreated, HLA-DR-negative cells . Other toxins such as diphtheria and pertussis toxins did not have the effect . The distribution of SEB and HLA-DR molecules was identical on the IFN-gamma-treated, HLA-DR-positive DJM-1 cells by confocal microscopy . Digital image processing analysis demonstrated that SEB induced a transient increase of intracellular calcium concentration only in the IFN-gamma-treated DJM-1 cells . Pretreatment of the IFN-gamma-treated DJM-1 cells with anti-major histocompatibility complex class II monoclonal antibody completely blocked the effect of SEB . Furthermore, ICAM-1 mRNA was detected in the IFN-gamma-pretreated, SEB-exposed normal keratinocytes by reverse transcriptase-polymerase chain reaction . Our results demonstrate that SEB binds to keratinocytes, presumably via major histocompatibility complex class II molecules such as HLA-DR, triggers calcium mobilization, and induces the synthesis of ICAM-1 molecules . We speculate that, in various cutaneous disorders, SEB penetrates the epidermis and interacts with HLA-DR-positive keratinocytes to upregulate ICAM-1 expression, thus modulating the course of the inflammatory process.

J Bacteriol, 1995 Oct, 177(19), 5723 - 5
Molecular characterization and functional analysis of the major autolysin of Staphylococcus aureus 8325/4; Foster SJ; The gene encoding the major autolysin of Staphylococcus aureus 8325/4 has been cloned, sequenced, and insertionally inactivated . The three-domain, 137,384-Da protein has a C-terminal glucosaminidase active site and is involved in cell separation, generalized cell lysis, and release of wall material at the cell surface . Expression occurs throughout growth and is stimulated by low temperatures and in the presence of 1 M NaCl.

Infect Immun, 1995 Oct, 63(10), 4121 - 9
Panton-Valentine leucocidin and gamma-hemolysin from Staphylococcus aureus ATCC 49775 are encoded by distinct genetic loci and have different biological activities; Prevost G et al.; Staphylococcus aureus ATCC 49775 produces three proteins recognized by affinity-purified antibodies against the S component of Panton-Valentine leucocidin (LukS-PV) and two proteins recognized by affinity-purified antibodies against the F component of this toxin (LukF-PV) . Purification of these proteins and cloning of the corresponding genes provided evidence for the presence of two loci . The first one, encoding Panton-Valentine leucocidin, consisted of two cotranscribed open reading frames, lukS-PV and lukF-PV, coding the class S and the class F components, respectively . The second one coded for a gamma-hemolysin and consisted of two transcription units, the first one encoding an HlgA-like protein, a class S component, and the second one encoding two cotranscribed open reading frames identical to HlgC and HlgB, class S and class F components, respectively, from gamma-hemolysin from the reference strain Smith 5R . It appears that the Panton-Valentine leucocidin from S . aureus ATCC 49775 (V8 strain) should not be confused with leucocidin from ATCC 27733 (another isolate of V8 strain), which had 95% identity with HlgC and HlgB from gamma-hemolysin . The cosecretion of these five proteins led to six possible synergistic combinations between F and S components . Two of these combinations (LukS-PV-LukF-PV and HlgA-LukF-PV) had dermonecrotic activity on rabbit skin, and all six were leukocytolytic on glass-adsorbed leukocytes . Only three were hemolytic on rabbit erythrocytes, the two gamma-hemolysin combinations and the combination LukF-PV-HlgA.

Infect Immun, 1995 Oct, 63(10), 3816 - 9
Differential effects of monoclonal antibody blockade of adhesion molecules on in vivo susceptibility to soft tissue infection; Garcia N et al.; Leukocyte adherence to endothelial cells has been implicated in the pathogenesis of microvascular injury as well as in host defense against various infectious microorganisms . Administration of monoclonal antibodies directed against the beta chain of the leukocyte integrins inhibits leukocyte-endothelial-cell adherence and has been reported to modulate ischemia-reperfusion and inflammatory injury . However, such inhibition of adhesion molecule function adversely affects resistance to infection . The following studies were carried out to determine whether monoclonal antibodies to other adhesion molecules, including L-selectin (CD62L), and CD11a (the alpha chain of LFA-1), also increase susceptibility to infection . New Zealand White rabbits were shaved and given subcutaneous injections on their dorsa with 10(9) CFU of Staphylococcus aureus ATCC 25923 at two sites and with 10(8) CFU at two sites . A second set of rabbits were given subcutaneous injections with 10(8) CFU of P . aeruginosa ATCC 27853 at two sites and with 10(7) CFUs at two sites . The animals were monitored for 1 week . There were three blinded experimental groups: controls given saline and two groups given blocking monoclonal antibodies to either L-selectin (Dreg-200) or CD11a (R7.1) . In contrast to monoclonal antibodies to CD18, none of the monoclonal antibodies significantly increased the risk of abscess formation by S . aureus, although inhibition of CD11a increased the rate of abscess formation by P . aeruginosa.

Kansenshogaku Zasshi, 1995 Oct, 69(10), 1126 - 34
{Appearance of antibacterial activity of oxacillin against methicillin resistant Staphylococcus aureus (MRSA) in the presence of catechin}; Takahashi O et al.; We previously reported that tea catechin shows bactericidal activity against various bacteria including methicillin resistant Staphylococcus aureus (MRSA) and that bactericidal catechin damages the lipid bilayer of bacterial cell membranes . Here we describe that oxacillin (MPIPC) shows antibacterial activity against MRSA in the presence of catechin below MIC . Twenty clinical isolates of MRSA were examined by a cup method . In the absence of catechin, MPIPC even at a concentration of 40 micrograms/ml did not show antibacterial activity against all isolates of MRSA . However, when catechin below MIC (25-100 micrograms/ml) was mixed with the agar medium, MPIPC (5-12.5 micrograms/ml) showed antibacterial activity against all MRSA isolates . By counting the numbers of viable bacteria in a broth culture, only MPIPC (5 micrograms/ml) or catechin (100 micrograms/ml) showed similar growth curves to the control . But addition of both MPIPC and catechin reduced the number of viable bacteria to 1/100-1/10000 after 24 hours of cultures . Besides MPIPC, in the presence of catechin below MIC methicillin (12.5 micrograms/ml), aminobenzylpenicillin (32 micrograms/ml), tetracycline (2.5 micrograms/ml), and chloramphenicol (12.5 micrograms/ml) showed antibacterial activities against multiple drug resistant MRSA to antibiotics mentioned above . These findings suggest a possible use of catechin in the treatment of MRSA infection.

J Biol Chem, 1995 Sep 29, 270(39), 23072 - 6
Interactions between residues in staphylococcal alpha-hemolysin revealed by reversion mutagenesis; Panchal RG et al.; alpha-Hemolysin (alpha HL), a pore-forming polypeptide of 293 amino acids, is secreted by Staphylococcus aureus as a water-soluble monomer . Residues that play key roles in the formation of functional heptameric pores on rabbit red blood cells (rRBC) have been identified previously by site-directed mutagenesis . alpha HL-H35N, in which the histidine at position 35 of the wild-type sequence is replaced with asparagine, is nonlytic and is arrested in assembly as a heptameric prepore . In this study, second-site revertants of H35N that have the ability to lyse rRBC were generated by error-prone PCR under conditions designed to produce single base changes . The analysis of 22 revertants revealed new codons clustered predominantly in three distinct regions of the H35N gene . One cluster includes amino acids 107-111 (four revertants) and another residues 144-155 (five revertants) . These two clusters flank the central glycine-rich loop of alpha HL, which previously has been implicated in formation of the transmembrane channel, and encompass residues Lys-110 and Asp-152 that, like His-35, are crucial for lytic activity . The third cluster lies in the region spanning amino acids 217-228 (eight revertants), a region previously unexplored by mutagenesis . Single revertants were found at amino acid positions 84 and 169 . When compared with H35N, the heptameric prepores formed by the revertants underwent more rapid conversion to fully assembled pores, as determined by conformational analysis by limited proteolysis . The rate of conversion to the fully assembled pore was strongly correlated with hemolytic activity . Previous work has suggested that the N terminus of alpha HL and the central loop cooperate in the final step of assembly . The present study suggests that the key N-terminal residue His-35 operates in conjunction with residues flanking the loop and C-terminal residues in the region 217-228 . Hence, reversion mutagenesis extends the linear analysis that has been provided by direct point mutagenesis.

J Biol Chem, 1995 Sep 29, 270(39), 23065 - 71
Key residues for membrane binding, oligomerization, and pore forming activity of staphylococcal alpha-hemolysin identified by cysteine scanning mutagenesis and targeted chemical modification; Walker B et al.; The alpha-hemolysin (alpha HL) polypeptide is secreted by Staphylococcus aureus as a water-soluble monomer that assembles into lipid bilayers to form cylindrical heptameric pores 1-2 nm in effective internal diameter . We have individually replaced each charged residue (79 of 293 amino acids) and four neutral residues in alpha HL with cysteine, which is not found in the wild-type protein . The properties of these mutants have been examined before and after modification with the 450-Da dianionic sulfhydryl reagent 4-acetamido-4'-((iodoacetyl)amino)stilbene-2,2'-disulfonate (IASD) . This modification was highly informative as 28 of 83 modified polypeptides showed substantially reduced pore forming activity on rabbit erythrocytes (rRBC), while only five of the unmodified cysteine mutants were markedly affected . Through detailed examination of the phenotypes of the mutant and modified hemolysins, we have pinpointed residues and regions in the alpha HL polypeptide chain that are important for binding to rRBC, oligomer formation and pore activity . Residues in both the N-terminal (Arg-66 and Glu-70) and C-terminal (Arg-200, Asp-254, Asp-255, and Asp-276) thirds of the protein are implicated in binding to cells . The His-35 replacement mutant modified with IASD was the only polypeptide in this study that failed to form SDS-resistant oligomers on rRBC . Altered hemolysins that formed oligomers but failed to lyse rRBC represented the most common defect . These alterations were clustered in the central glycine-rich loop, which has previously been implicated as a component of the lumen of the membrane-spanning channel, and in the regions flanking the loop . Alterations in mutant and modified hemolysins with the same defect were also scattered between the N terminus and His-48, in keeping with previous suggestions that an N-terminal segment and the central loop cooperate in the final step of pore assembly.

MMWR Recomm Rep, 1995 Sep 22, 44(RR-12), 1 - 13
Recommendations for preventing the spread of vancomycin resistance . Recommendations of the Hospital Infection Control Practices Advisory Committee (HICPAC); Purification to homogeneity of a novel human B cell differentiation factor; Mount Sinai Medical Center, Division of Clinical Immunology, New York, NY 10029, USABy immunization with a partially purified form of a novel human B cell differentiation factor, 446-BCDF, we generated a series of mAbs that inhibit the functional activity of this cytokine (induction of Ig secretion by Staphylococcus aureus Cowan I strain organisms-activated B cells) . Two mAbs, 929 and 204, were shown to be specific for 446-BCDF in that they failed to inhibit B cell differentiation in response to other known cytokines (IL-2, IL-6) or mitogens (PWM) . More importantly, passage of crude 446-BCDF over a mAb 929-Sepharose 4B column resulted in the depletion of 446-BCDF activity, which could be recovered after elution with 0.1 M glycine, pH 2.8 . Finally, TCA precipitation of column-eluted fractions was run on SDS-PAGE . Two bands corresponding to the previously described m.w . range of 446-BCDF were detected in the eluate, but not in the effluent column fractions . Furthermore, protein eluted from these bands resolved with nondenaturing gels also demonstrated 446-BCDF activity . Thus, together these data further support the existence of a novel human B cell differentiation factor that has been purified to homogeneity.

EMBO J, 1995 Sep 15, 14(18), 4569 - 77
Activation of alpha-toxin translation in Staphylococcus aureus by the trans-encoded antisense RNA, RNAIII; Morfeldt E et al.; The synthesis of virulence factors in Staphylococcus aureus is controlled by a regulatory RNA molecule, RNAIII, encoded by the agr locus . Transcription of genes coding for secreted toxins and enzymes is stimulated, while transcription of cell-surface protein genes is repressed by RNAIII . In the case of staphylococcal alpha-toxin, RNAIII also seems to stimulate translation by an independent mechanism . In this report we show that in a mutant lacking RNAIII the rate of alpha-toxin (hla) production relative to the cellular concentration of hla mRNA was reduced 10-fold as compared with the wild-type strain . A 75% complementarity between the 5' end of RNAIII and the 5' untranslated region of the hla transcript suggests a direct interaction between the RNAs . A complex of RNAIII and hla mRNA was demonstrated in extracts of total RNA from the wild-type strain, and also with in vitro synthesized RNAs . Ribonuclease T1 digestion experiments revealed that the ribosome binding site of the hla transcript is blocked by intramolecular base-pairing . Hybridization with RNAIII prevents this intramolecular base-pairing and makes the hla mRNA accessible for translation initiation . This is, to our knowledge, the first example of an 'antisense RNA' that stimulates translation of the target mRNA.

J Biol Chem, 1995 Sep 15, 270(37), 21457 - 60
Staphylococcus aureus expresses a major histocompatibility complex class II analog; Jonsson K et al.; Staphylococcus aureus expresses various surface proteins which specifically recognize and bind to different host molecules . We have previously identified a bacterial protein that exhibits a broad specificity and binds to several mammalian extracellular proteins . The gene encoding this bacterial component has now been cloned and sequenced . The deduced protein consists predominantly of six repeated domains of 110 residues . Each of the repeated domains contain a subdomain of 31 residues that share striking sequence homology with a segment in the peptide binding groove of the beta chain of the major histocompatibility complex (MHC) class II proteins from different mammalian species . The purified recombinant bacterial protein bound several mammalian proteins, including recombinant osteopontin, suggesting a protein-protein interaction and also specifically recognized a 15-amino acid residue synthetic peptide . Taken together, these results suggest that the bacterial protein resembles mammalian MHC class II molecules with respect to both sequence similarities and peptide binding capabilities.

Dtsch Med Wochenschr, 1995 Sep 8, 120(36), 1191 - 6
{Infectious endocarditis of native and prosthetic valves . A comparative analysis based on 155 cases}; Morguet AJ et al.; OBJECTIVE: In a retrospective study, we compared the characteristics of native valve and prosthetic valve endocarditis . PATIENTS AND METHODS: All 155 cases of left-sided infective endocarditis in 142 patients admitted at our institution between 1986 and 1992 were analyzed based on their medical records . Native valve endocarditis was found in 119 cases (74 men, 45 women; median age 55, range 29 to 80 years), prosthetic valve endocarditis in 36 cases (11 men, 25 women; median age 63, range 38 to 80 years; 29 cases of late infection) . RESULTS: There were more older (P < 0.0005) and more female (P = 0.001) patients with prosthetic valve endocarditis . Most frequently, native aortic valves (53.8%) or mitral prostheses (55.6%) were solely involved . In both groups, Staphylococcus epidermidis was the most frequent isolate followed by Staphylococcus aureus and Enterococci . Symptoms and clinical findings were similar in both groups . In all cases since July 1989, transthoracic echocardiography was suggestive of endocarditis in native valves in 63.9% and in prostheses in 29.2% (P = 0.004), combined with the transesophageal approach in 91.7 and 91.8%, respectively (not significant) . Embolism and surgical intervention were about equally frequent in both groups . The in-hospital mortality was 19.3% in native valves and 22.2% in prostheses (not significant) . CONCLUSION: Native valve and late prosthetic valve endocarditis are now similar in the spectrum of causative microorganisms, echocardiographic diagnostics, clinical course and prognosis.

J Antimicrob Chemother, 1995 Sep, 36(3), 551 - 5
Efficacy of quinolones in preventing Staphylococcus aureus-induced abscess in mice; Cohen MA et al.; The new fluoroquinolones clinafloxacin, CI-990 (PD 131112/PD 131628), sparfloxacin, and PD 138312 were bactericidal against Staphylococcus aureus with MICs > or = 4-fold lower than ciprofloxacin values . In a murine subcutaneous abscess model (subcutaneous/oral dosing) the new drugs displayed protective activities against five strains which were generally higher (up to 19-fold) than ciprofloxacin values.

Afr J Med Med Sci, 1995 Sep, 24(3), 243 - 8
Neonatal seizures in Nigerian infants; Asindi AA et al.; Nigerian newborns presenting with convulsion in University of Calabar Teaching Hospital, Calabar during the period January 1989 to December 1990 were prospectively studied to determine the aetiology and pattern of their seizures . There were 60 patients representing 4% of admissions into the Newborn Unit during the period . Birth asphyxia, infections and hypoglycaemia were the important identifiable aetiological factors which operated either singly (48% of cases) or in concert (in another 48%) of the infants . Detectable infections included meningitis and septicaemia caused predominantly by coliforms and Staphylococcus aureus . Hypocalcaemia and electrolyte imbalance did not feature . There was an unusually high prevalence (63% of cases) of the generalised type of seizures probably due to the high frequency of mixed aetiology . The mortality rate of 50% encountered appears to be related to the underlying aetiology and prematurity . Detectable caused of neonatal seizures in our environment appear to be potentially preventable by improved obstetric and neonatal care . There is dire need also to provide modern facilities for investigating newborn seizures in order to improve upon the diagnostic yieldPIP: During 1989-1990 in Nigeria, 60 newborns were admitted to the Special Care Baby Unit of the University of Calabar Teaching Hospital for seizures, representing 4.1% of admissions . 11.7% of the newborns with seizures were preterm . 48.3% had more than 1 detectable cause of the seizures . The leading cause of seizures was asphyxia (36 newborns) followed by infections (16), especially meningitis (8) . Coliforms and Staphylococcus aureus were the primary etiologic agents for the infections . Among the 36 newborns with asphyxia, 13 also suffered from hypoglycemia, 13 also suffered from infection, and 5 also suffered from infection and hypoglycemia . 63% of the newborns had generalized seizures, which may be associated with the mixed etiology in 48% of the cases . 50% of the 60 newborns presenting with seizures died . Newborns whose seizures were associated with infection had the highest fatality rate (50%) . The mortality rate was lowest in infants whose seizures occurred during the first day of life (36.7% vs . 86% 4-7 days after birth) . The causes of neonatal seizures in these newborns can be prevented by improving obstetric and neonatal care . Pediatricians need access to modern diagnostic and therapeutic facilities to improve the accuracy of diagnoses in cases of newborn seizures . Health workers need to increase health education activities geared to prenatal patients .

Zentralbl Veterinarmed B, 1995 Sep, 42(7), 435 - 42
Inflammation in the bovine teat cistern induced by Staphylococcus aureus; Persson K et al.; The inflammatory response, characterized by the accumulation of leukocytes, bovine serum albumin and the lysosomal enzyme N-acetyl-beta-D-glucosaminidase, was studied after inoculation of either 5 x 10(5) colony-forming units (CFU) or 2 x 10(2) CFU of the Staphylococcus aureus strain SA 14391 into teat cisterns of dry cows after surgical closure of the passage between the teat and udder cisterns . Teat cistern samples were taken before, and twice daily for 7 days after, inoculation of the bacteria . Infusion of sterile saline constituted a control . Persistent infections occurred in all teats inoculated with the higher dose (5 x 10(5) CFU) of bacteria, and a prominent inflammatory response was elicited . Marked differences were observed in leukocyte migration patterns between different cows, and a cyclic influx of leukocytes was evident . Inoculation of the lower dose (2 x 10(2) CFU) of bacteria did not result in a persistent infection, and only a slight inflammatory response was observed . The results indicate that the bovine teat tissues are capable of mounting a strong local inflammatory response to S . aureus infection . A large number of leukocytes invaded the teat, but, despite their numbers, they were unable to subdue the infection, except when the bacterial count was low.

Mol Microbiol, 1995 Sep, 17(6), 1143 - 52
Adhesion properties of mutants of Staphylococcus aureus defective in fibronectin-binding proteins and studies on the expression of fnb genes; Greene C et al.; Staphylococcus aureus 8325-4 has the potential to express two distinct cell wall-associated fibronectin-binding proteins called FnBPA and FnBPB . In order to test if both proteins are expressed in S . aureus and if both are required for promoting bacterial adhesion to fibronectin-coated surfaces, insertion mutations were isolated in each gene . A DNA fragment encoding tetracycline resistance was inserted into fnbA and a fragment encoding erythromycin resistance was inserted into fnbB . A double fnbAfnbB mutant was also constructed . The fnbA and fnbB single mutants showed no significant reduction in their adhesion to polymethylmethacrylate coverslips that had been coated in vitro with fibronectin . However, the double mutant was completely defective in adhesion . Monospecific antibodies directed against the non-conserved N-terminal regions of both proteins confirmed the lack of expression of FnBPs in the mutant strains . Wild-type fnbA and fnbB genes cloned seperately on a multicopy plasmid were each able to restore fully the adhesion-defective phenotype of the 8325-4 fnbAfnbB mutant . This demonstrates that both fnb genes are expressed in S . aureus and that both contribute to the ability of strain 8325-4 to adhere to fibronectin-coated surfaces . The double mutant was also defective in adhesion to coverslips that had been removed from tissue cages implanted subcutaneously in guinea-pigs, which suggests that fibronectin is important in promoting attachment of S . aureus to biomaterial in vivo.

Cutis, 1995 Sep, 56(3), 158 - 60
Botryomycosis in a patient with acquired immunodeficiency syndrome; Salvemini JN et al.; Botryomycosis is a bacterial infection of either the skin alone or in combination with visceral organs . It resembles a deep fungal infection . A histologic evaluation of cutaneous lesions reveals the characteristic Splendore-Hoepple phenomena and assists with management . Patients with acquired immunodeficiency syndrome (AIDS) tend to have uncharacteristic lesions resembling common conditions such as prurigo nodularis and lichen simplex chronicus . Diagnosis in these cases can be challenging . We report the case of a patient with AIDS who was successfully treated with Augmentin (amoxicillin, clavulanate potassium) . Complete resolution of the lesion occurred after the causative agent, Staphylococcus aureus, was identified.

Immunopharmacology, 1995 Sep, 30(3), 231 - 6
Modulatory effect of plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone) on macrophage functions in BALB/c mice . I . Potentiation of macrophage bactericidal activity; Abdul KM et al.; The modulatory ability of plumbagin, a natural product from Plumbago zeylanica, was studied on peritoneal macrophages of BALB/c mice . The macrophage functions evaluated were bactericidal activity, hydrogen peroxide and superoxide anion release . The bactericidal capacity of in vivo plumbagin-treated mouse macrophages was estimated against Staphylococcus aureus . In low doses plumbagin exerted a constant increase in bactericidal activity throughout the study period whereas with a high dose a higher response was observed up to six weeks . But in the next two weeks a considerable decline in the bactericidal activity was noticed compared to low dose . Plumbagin was also seen to exert a similar response on oxygen radical release by macrophages in vivo showing a clear correlation between oxygen radical release and the bactericidal activity . The data indicate that plumbagin augments the macrophage bactericidal activity by potentiating the oxyradical release at low concentration whereas at the higher concentration it has inhibitory activity.

J Dairy Sci, 1995 Sep, 78(9), 2083 - 5
Segregation or use of separate milking units for cows infected with Staphylococcus aureus: effects on prevalence of infection and bulk tank somatic cell count; Wilson DJ et al.; Dairy herds (n = 76) with an initial prevalence of Staphylococcus aureus IMI > or = 10% were included in this study . Criteria were that herds did not change teat dipping or dry cow treatment practices, did not segregate cows that were positive for S . aureus at the initial visit, and did not cull > 50% of cows found to be positive on the initial visit . During a follow-up period (6 to 24 mo), segregation or separate milking of cows that were positive for S . aureus reduced prevalence from 29.5 to 16.3% and bulk tank SCC from 600,000 to 345,000/ml . Prevalence of S . aureus mastitis was unchanged for herds that did not segregate cows with S . aureus, 22.5 to 20.2%, and the reduction in SCC from 698,000 to 484,000 for nonsegregated herds was also smaller . Segregation of cows that were known to be positive for S . aureus is an effective mastitis control practice.

J Dairy Sci, 1995 Sep, 78(9), 1937 - 44
Differences in bovine lymphocyte antigen associations between immune responsiveness and risk of disease following intramammary infection with Staphylococcus aureus; Mallard BA et al.; This study evaluated the relationships between immune response, disease resistance, and bovine leukocyte antigens, BoLA, in Holstein cows following intramammary challenge with Staphylococcus aureus . This investigation was to determine whether immune responsiveness differed between these cows and whether differences were related to expression of class I BoLA antigens, which might explain the increased resistance or susceptibility to S . aureus mastitis . Antibody responses to S . aureus in milk and serum, total IgG1 in milk, and blastogenic responses with and without concanavalin A were evaluated . The CA42 allele, previously associated with increased risk of infection, was relatively uninformative for the immune response parameters examined . Other alleles, such as W3, were associated with higher milk antibody responses to S . aureus and higher milk IgG1 postchallenge . Alleles W7, W4, and W26 were associated with lower milk IgG1 and lower antibody titers in serum postchallenge . The association reported between allele CA42 and increased risk of S . aureus mastitis did not relate to lower antibody or blastogenic responses by these cows; however, indicators of innate resistance were not examined . In addition, the different associations detected between milk and serum emphasize the importance of considering mucosal immunity and unique immunological compartments when searching for relevant genetic markers of disease resistance.

J Dairy Sci, 1995 Sep, 78(9), 1932 - 6
Efficacy of a Propionibacterium acnes immunostimulant for treatment of chronic Staphylococcus aureus mastitis; Dinsmore RP et al.; This study determined the efficacy of treatment of chronic Staphylococcus aureus IMI of lactating dairy cows with a biological response modifier consisting of a killed preparation of Propionibacterium acnes . Fifty-one lactating dairy cows with chronic S . aureus IMI on two commercial dairies were enrolled in a controlled, randomized field trial . Twenty-four cows received 1 ml of the immunostimulant twice weekly for 4 wk, and the remainder of the cows served as untreated controls . Quarter milk samples were collected for SCC from all cows at 3 and 6 wk after initiation of treatment and for culture at 6 wk after initiation of treatment . If samples were negative for S . aureus at 6 wk, cows were sampled again for culture 1 wk later for 2 successive d before being considered cured . For the cows treated with immunostimulant, 16.7% (36.7% of infected quarters) were cured of S . aureus IMI; for the untreated group, 11.1% of cows (32.2% of quarters) were cured . The difference was not statistically significant . Immunostimulant treatment had no effect on quarter SCC in infected quarters overall, but, in those quarters that were cured of S . aureus infection, quarter SCC of treated cows tended to be lower than SCC of control cows.

J Dermatol, 1995 Sep, 22(9), 673 - 6
Necrotizing fasciitis in association with hyperimmunoglobulin E syndrome; Misago N et al.; A case of necrotizing fasciitis in association with hyperimmunoglobulin E (HIE) syndrome is reported . The patient was a 17-year-old Japanese boy with a clinical history of recurrent skin and pulmonary infections and eczematoid dermatitis, markedly elevated serum levels of IgE, and coarse facies . He had a gangrenous swelling on the lower abdominal wall, and his general condition was poor with high fever . The involved site was accompanied by subcutaneous gas; the culture of the pus of the lesion grew anaerobes without mixed growth of Staphylococcus aureus . Exhaustive debridement of necrotic fascia, which extended much farther than the gangrenous area, and administration of antibiotics had a curative effect on the gangrenous soft tissue infection . To the best of the authors' knowledge, this is the first published case of necrotizing fasciitis in association with HIE syndrome.

J Biolumin Chemilumin, 1995 Sep-Oct, 10(5), 291 - 9
Enumeration of phagocytosed Staphylococcus aureus inside cells of the mouse macrophage cell line J774 by bioluminescence, fluorescence and viable counting techniques; Sunderland J et al.; In order to quantify intracellular Staphylococcus aureus within a macrophage-like cell line by a bioluminescence technique, the mouse cell line J774 and opsonized Staphylococcus aureus were incubated together to allow phagocytosis to occur . Experiments using UV microscopy and fluorescent stained S . aureus were performed to determine an estimate of the mean intracellular bacterial numbers . For enumeration of intracellular bacteria by a bioluminescence technique, extracellular bacteria were removed by washing, the macrophages lysed mechanically and osmotically and treated with apyrase to remove somatic ATP . Bacterial cells were washed and the intracellular ATP measured by firefly luciferase bioluminescence in a luminometer . This new method of enumerating intracellular bacteria was compared to the conventional method of viable counts and found to correlate (r = 0.78) . The bioluminescence assay developed was found to be a relatively rapid alternative method to the techniques currently used to enumerate intracellular bacteria and could prove advantageous in studies of intracellular killing and effects of antimicrobial agents on intracellular pathogens.

Vet Immunol Immunopathol, 1995 Sep, 48(1-2), 65 - 76
Up-regulation of IL-2 receptor alpha and MHC class II expression on lymphocyte subpopulations from bovine leukemia virus infected lymphocytotic cows; Stone DM et al.; Infection with bovine leukemia virus (BLV) leads to a persistent lymphocytosis (PL) characterized by a marked increase in circulating B lymphocytes that express the orthologue of CD5 . To gain insight into the factors accounting for lymphocytosis, experiments were conducted to determine the functional activation status of lymphocytes from BLV seronegative and BLV infected aleukemic cows with PL . Stimulation with the B lymphocyte mitogen Staphylococcus aureus Cowan strain I (SAC), recombinant human interleukin-2 (rIL-2), or pokeweed mitogen (PWM), a T lymphocyte-dependent B lymphocyte mitogen, revealed differences in the pattern of expression of IL-2 receptor alpha (IL-2R alpha) and major histocompatibility (MHC) class II molecules on B and T lymphocytes from uninfected and BLV infected PL cows . rIL-2 induced expression of IL-R alpha on B lymphocytes from PL cows but not B lymphocytes from BLV seronegative cows . SAC alone, or in combination with rIL-2, had no effect on B lymphocytes from BLV seronegative cows . However, rIL-2 alone or in combination with SAC induced expression of IL-2R alpha on B lymphocytes from PL cows . PWM stimulated expression of IL-2R alpha on bovine B lymphocytes regardless of BLV status, and induced a significantly higher level of expression on B lymphocytes from PL cows . Mitogens and rIL-2 had a similar stimulatory effect on induction of IL-2R alpha expression on CD4 T lymphocytes regardless of BLV status . Only PWM induced expression of IL-2R alpha on bovine CD8 T lymphocytes and induced a significantly higher level of expression on this T lymphocyte subset from PL cows . Examination of freshly isolated B lymphocytes from PL cows revealed increased spontaneous expression of the MHC class II molecule compared to B lymphocytes from control cows . None of the culture conditions examined induced MHC-II expression on CD4 and CD8 T lymphocytes from BLV seronegative cows . In contrast, SAC+IL-2 and PWM induced MHC-II expression on CD4 and CD8 T lymphocytes from BLV infected PL cows, resulting in a significantly greater proportions of these lymphocyte subsets expressing this molecule compared to CD4 and CD8 T lymphocytes from control cows . The data indicate that infection with BLV affects the response of B and T lymphocytes to signals of activation, up-regulating the expression of surface molecules involved in both direct contact and cytokine-mediated T lymphocyte-dependent B lymphocyte activation.

J Int Med Res, 1995 Sep-Oct, 23(5), 328 - 34
Sensitivity of Staphylococcus aureus, isolated from skin infections in 1994, to 19 antimicrobial agents; Nishijima S et al.; The most common pathogen causing skin infections is Staphylococcus aureus and the incidence of multiply resistant strains of S . aureus has been increasing . The in vitro susceptibility of 130 isolates of S . aureus to 19 antimicrobial agents: ampicillin (ABPC), methicillin, cefaclor, cefpodoxime proxetil, gentamicin, erythromycin, clindamycin, minocycline, vancomycin, fusidic acid, norfloxacin, ofloxacin, enoxacin, ciprofloxacin, lomefloxacin, tosufloxacin, sparfloxacin, nadifloxacin and grepafloxacin, was evaluated by agar dilution tests . The S . aureus isolates were isolated from 130 patients with skin infections in 1994 . The proportion of methicillin-resistant S . aureus isolates among the strains isolated was 19.2% . The concentration needed to inhibit 50% of the isolates was 3.13 mg/ml or less for all of the drugs, but the concentration needed to inhibit 90% of isolates was over 12.5 micrograms/ml, except in the cases of minocycline, vancomycin, fusidic acid, tosufloxacin and nadifloxacin . Tosufloxacin and nadifloxacin had the lowest minimum inhibitory concentrations . None of the S . aureus strains was resistant to nadifloxacin.

Zh Mikrobiol Epidemiol Immunobiol, 1995 Sep-Oct, (5), 64 - 7
{The epidemiological surveillance of hospital infections linked to methicillin-resistant Staphylococcus aureus}; Musina LT et al.; As revealed in the realization of the epidemiological surveillance of hospital infections caused by methicillin-resistant S . aureus (MRSA) in different types of hospital, MRSA strains causing purulent inflammatory diseases belong to different clones . The complex marking of MRSA made it possible to determine the presence of the same clone in different hospitals and to detect the outbreaks of hospital infections caused by different clones of MRSA in one hospital . It was found necessary to supplement the commercial international phage-typing set with pages permitting the detection of the specific system of restriction-modification in MRSA.

Zh Mikrobiol Epidemiol Immunobiol, 1995 Sep-Oct, (5), 19 - 23
{The effect of individual serum proteins and native blood serum on the adhesion of Staphylococcus aureus to cells in vitro}; Eropkina EM et al.; Blood serum and its components were found to produce an antiadhesive effect which inhibited the attachment of S.aureus to cells HEp-2 and immortalized astrocytes . Normal and antistaphylococcal immunoglobulins exhibited the highest activity, inhibiting the process of bacterial adhesion in a serum-free medium . The antiadhesive activity level of native serum was considerably lower and constituted 4% of that of normal immunoglobulin and 85% of that of albumin . In spite of pronounced inhibiting action of normal immunoglobulin and albumin, their dissolution in serum did not increase its antiadhesive activity.

Chemotherapy, 1995 Sep-Oct, 41(5), 412 - 9
A comparative study between teicoplanin alone and flucloxacillin, plus or minus fusidic acid, in the treatment of serious infections caused by methicillin-susceptible gram-positive bacteria; Mehtar S et al.; A randomized trial compared teicoplanin alone against flucloxacillin, with or without fusidic acid, in the treatment of serious gram-positive infections . The majority of infections involved Staphylococcus aureus or Staphylococcus epidermidis, methicillin-resistant organisms were excluded . A total of 56 patients were evaluable for efficacy, with no significant differences between treatment groups . Clinical success (cure + improvement) was achieved in 24/27 patients on teicoplanin (89%), 16 on flucloxacillin (100%) and 8/9 receiving flucloxacillin/fusidic acid (89%) . Adverse events occurred in 21% of patients (7 on teicoplanin and 6 receiving flucloxacillin +/- fusidic acid) . All such events resolved spontaneously or following appropriate management . It is concluded that teicoplanin monotherapy, 400 mg once daily, shows similar efficacy and tolerability to multiple daily doses of flucloxacillin, with or without fusidic acid, in the treatment of methicillin-susceptible serious gram-positive infection.

Support Care Cancer, 1995 Sep, 3(5), 324 - 6
Immune-complex glomerulonephritis associated with Staphylococcus aureus infection of a totally implantable venous device; Pulik M et al.; We report a previously unrecognized complication of totally implanted subcutaneous ports . A patient with a totally implantable central venous device developed a septic syndrome . Blood and injection-reservoir cultures grew Staphylococcus aureus . There was no evidence of endocarditis . The port was removed but acute oliguric renal failure developed . A percutaneous renal biopsy showed acute diffuse proliferative glomerulonephritis . There was no extracapillary crescent . On immunofluorescence study, capillary wall granular deposits stained brightly for IgG and C3 . These findings were thought to be consistent with infection-associated immune-complex glomerulonephritis.

Support Care Cancer, 1995 Sep, 3(5), 297 - 300
Totally implanted catheters to reduce catheter-related infections in patients receiving interleukin-2: a 2-year experience; Escudier B et al.; A high incidence of bacterial infections has been previously reported during interleukin-2 (IL-2) treatment, mainly due to catheter-related infections . Antibiotic prophylaxis has been successfully used to decrease such infections . The goal of this study was to evaluate an alternative way to reduce catheter-related infections in IL-2-treated patients by the use of totally implanted catheters . A total of 74 patients with metastatic renal cell carcinoma, referred to our institution to receive IL-2 from March 1989 to July 1991, were included in this prospective study . IL-2 was given on a 2-days-a-week schedule (24 x 10(6) IU m-2 day-1) either alone (41 patients) or in association with interferon gamma (33 patients) . All these patients were prospectively evaluated for fever, bacteremia and line-site infection . Seven patients (9.5%) had one (2 patients) or more (5 patients) positive blood cultures with Staphylococcus aureus . Antibiotics were used only in 5 patients, and the catheter had to be removed in only 2 of these patients . In the other patients, no further infection developed despite the lack of antibiotics . Moreover, 9 patients had positive blood cultures with Staphylococcus epidermidis (1.9% of total number of blood cultures) . In conclusion, a totally implanted catheter appears to reduce the incidence of infections in IL-2-treated patients, at least on a 2-days-a-week schedule.

Biosci Biotechnol Biochem, 1995 Sep, 59(9), 1786 - 9
Substitution of lysine for arginine in the N-terminal 217th amino acid residue of the H gamma II of Staphylococcal gamma-hemolysin lowers the activity of the toxin; Sudo K et al.; The staphylococcal toxin gamma-hemolysin consists of two protein components, LukF and H gamma II . Staphylococcus aureus P83 was found to have five components, LukF, LukF-PV, LukM, LukS, and H gamma II for leukocidin or gamma-hemolysin . H gamma II of S . aureus P83 was demonstrated to be a naturally-occurring analogous molecule of H gamma II {H gamma II(P83)}, in which the 217th arginine residue was replaced by lysine . The H gamma II(P83) showed about 50% of the hemolytic activity of normal H gamma II in the presence of LukF.

J Bacteriol, 1995 Sep, 177(17), 5116 - 21
Increased cell size and shortened peptidoglycan interpeptide bridge of NaCl-stressed Staphylococcus aureus and their reversal by glycine betaine; Vijaranakul U et al.; Staphylococcus aureus cells grown in a defined medium under conditions of high ionic stress (2.5 M NaCl) were significantly larger than cells grown under unstressed conditions, even though the cells grew much more slowly under stressed conditions . Analysis of the structure of peptidoglycan from stressed cells showed a shorter interpeptide bridge than in peptidoglycan from unstressed cells . Glycine betaine inclusion in the high-NaCl medium resulted in cells with sizes and interpeptide bridges similar to those of cells grown under unstressed conditions.

Am J Surg, 1995 Sep, 170(3), 271 - 6
Clinical effects of continuous microwave for postoperative septic wound treatment: a double-blind controlled trial; Korpan NN et al.; BACKGROUND: Continuous microwave (CM) has already been shown to be effective in treating various pathologic states . The aim of this trial was to study the curative effect of this new physical method on the course of postoperative suppurative and inflammatory processes in patients who underwent abdominal surgery . PATIENTS AND METHODS: In this study, 141 patients with postoperative purulent wounds (predominantly caused by pyogenic Staphylococcus aureus) were randomized into two groups: 71 patients received local CM therapy (group A), and the other 70 patients received a placebo treatment using a similar but ineffective device (group B, controls) . In this double-blind study, criteria for wound healing in both patient groups were evaluated . RESULTS: Results demonstrated that wound clearance was significantly accelerated in group A (treated with CM) compared with group B (controls): 5.6 +/- 0.6 versus 10.2 +/- 0.5 days (mean +/- standard deviation), respectively . Similarly, initial epithelization was significantly forced in group A compared with group B: 7.0 +/- 0.4 versus 12.8 +/- 0.6 days, respectively; and granulation appeared after 4.9 +/- 0.2 versus 8.7 +/- 0.4 days of postoperative treatment, respectively . Daily decrease of wound surface area was significantly higher in group A than in group B (7.1% versus 3.2%) . On the fifth postoperative day of treatment, the number of microorganisms was considerably lower (10(5) per gram of tissue) in patients treated with CM than in controls . CONCLUSIONS: The results of this controlled clinical trial suggest that low-intensity CM is an effective postoperative treatment of purulent wounds after abdominal surgery . Further investigations may elucidate the underlying mechanisms in detail and optimize the curative effects in surgical practice.

Chest, 1995 Sep, 108(3), 786 - 8
Bacteremic nosocomial pneumonia . A 7-year experience in one institution; Taylor GD et al.; STUDY OBJECT: To describe the epidemiology, microbiology, and outcome of nosocomial pneumonia with secondary bloodstream infection . DESIGN: Prospective cohort study . SETTING: Tertiary care Canadian teaching hospital . PATIENTS: Inpatients . MEASUREMENT: All inpatient blood cultures were concurrently monitored over an 89 month period . Following chart review, patients experiencing nosocomial bloodstream infection due to pneumonia were identified . A standardized definition of pneumonia was used . RESULTS: One hundred forty-nine episodes occurred in 145 patients, 0.66/1,000 hospital admissions, 8.4% of all nosocomial bloodstream infections . No change in rate occurred in the study period . Fifty-four percent of episodes developed in one of seven ICUs . Staphylococcus aureus was the most frequently identified etiologic organism (27%) . The ICU and non-ICU cases did not differ in etiology . No organism became more prevalent during the study period . Twenty percent of patients died within 1 week of first positive culture; episodes associated with Pseudomonas species had a much higher mortality rate (45%) than other infections (14%) (p = 0.002) . The ICU and non-ICU infections had a similar mortality rate . CONCLUSION: Pneumonia is an important cause of nosocomial bloodstream infection, but it is not increasing in frequency or changing in etiology in our institution . The ICUs are a major contributor to this problem but have the same case short-term mortality rate and microbial etiology as non-ICU cases . Cases associated with Pseudomonas have a much higher mortality rate.

Anesth Analg, 1995 Sep, 81(3), 555 - 8
Bactericidal activity of skin disinfectants on methicillin-resistant Staphylococcus aureus; Sakuragi T et al.; We studied bactericidal activity of 10% povidone-iodine, 0.5% chlorhexidine gluconate, and 0.5% chlorhexidine in 80% ethanol on four strains of methicillin-resistant and two strains of methicillin-susceptible Staphylococcus aureus . The pathogen was exposed to each of the disinfectants for 15, 30, 60, 120, and 240 s at room temperature . The inocula from these suspensions were cultured 72 h at 37 degrees C after the antimicrobial activity of the disinfectants in the suspensions was inactivated by 1:1000 dilution with neutralizer . No organism grew in any of the strains after exposure to 0.5% chlorhexidine in 80% ethanol . The 15-, 30-, and 60-s exposure to 10% povidone-iodine reduced the mean colony count by 55.2%, 91.2%, and 96.7%, respectively, and the exposures to 0.5% chlorhexidine gluconate reduced the mean colony count by 37.2%, 77.1%, and 93.3%, respectively . The difference in colony count between disinfectants was significant at 15- and 30-s exposures (P < 0.01 and 0.05, respectively) . The results suggest that bactericidal activity of 0.5% chlorhexidine in 80% ethanol is more potent and more rapid against methicillin-susceptible and methicillin-resistant strains of S . aureus.

J Med Microbiol, 1995 Sep, 43(3), 221 - 3
Microbiology of gastrostomy site wound infections in children; Brook I; Specimens of pus were obtained from gastrostomy site wound infections in 22 children . Polymicrobial flora was found in 21 of the 22 wounds . Aerobic or facultative bacteria only were isolated in eight (36%) instances and mixed aerobic-anaerobic flora were isolated from the other 14 (64%) wounds . A total of 102 bacterial isolates (57 aerobic and 45 anaerobic) and seven cultures of candida were obtained . The most frequent isolates were Escherichia coli (16 isolates) . Peptostreptococcus spp . (14), Enterococcus spp . (14), Bacteroides spp . (12) and Staphylococcus aureus (6) . Twenty-eight strains producing beta-lactamase were isolated from 16 (73%) patients . The presence of polymicrobial aerobic-anaerobic infection, and the isolation of E . coli and Bacteroides spp . were more frequent in wounds with gastric leakage than in wounds without gastric leakage (p < 0.05) . Bacteria similar to those isolated from the wound were also isolated from blood cultures from three patients-two isolates of E . coli and one each of B . fragilis and S . aureus . All patients received local therapy and 11 were treated with systemic antimicrobial agents . The polymicrobial aerobic-anaerobic flora of gastrostomy site wound infections, especially in association with gastric leakage, and the presence of beta-lactamase producers in most of these infections may have important implications for their management.

Int Arch Allergy Immunol, 1995 Sep, 108(1), 11 - 8
Isolation and partial characterization of eosinophil granule proteins in rats--eosinophil cationic protein and major basic protein; Watanabe M et al.; Peritoneal eosinophilia was induced in rats using Ascaris suum extract as an antigen, and characteristics of granule proteins of eosinophils collected from the peritoneal cavity were investigated . Peritoneal eosinophilia was induced by injection of the antigen solution into the peritoneal cavity of the immunized rats that had been orally administered with cyclophosphamide . Peritoneal cells were collected 48 h after injection of the antigen solution, incubated in plastic dishes, and nonadherent cells were collected as an eosinophil-rich fraction, from which granule proteins were extracted . Granule proteins were then purified by cation exchange chromatography, gel filtration, copper chelate affinity chromatography, and reverse-phase HPLC . Two distinct basic proteins of which molecular weights are 18- and 17-kD were obtained . Partial N-terminal amino acid sequence analysis revealed that the 18-kD protein and the 17-kD protein were homologous to the human eosinophil cationic protein (ECP) and the human and guinea pig major basic protein (MBP), respectively . Both the two proteins showed strong bactericidal activity against Escherichia coli and Staphylococcus aureus . These results indicate that rat eosinophils also possess ECP and MBP in their granules as well as human and guinea pig eosinophils.

Infect Immun, 1995 Sep, 63(9), 3634 - 41
Staphylococcus aureus induces platelet aggregation via a fibrinogen-dependent mechanism which is independent of principal platelet glycoprotein IIb/IIIa fibrinogen-binding domains; Bayer AS et al.; Platelet aggregation by bacteria is felt to play an important role in the pathogenesis of infective endocarditis . However, the mechanisms involved in bacterium-induced platelet aggregation are not well-defined . In the present study, we examined the mechanisms by which Staphylococcus aureus causes rabbit platelet aggregation in vitro . In normal plasma, the kinetics of S . aureus-induced platelet aggregation were rapid and biphasic . The onset and magnitude of aggregation phase 1 varied with the bacterium-platelet ratio, with maximal aggregation observed at a ratio of 5:1 . The onset of aggregation phase 2 was delayed in the presence of apyrase (an ADP hydrolase), suggesting that this later aggregation phase may be triggered by secreted ADP . The onset of aggregation phase 2 was delayed in the presence of prostaglandin I2-treated platelets, and this phase was absent when paraformaldehyde-fixed platelets were used, implicating platelet activation in this process . Platelet aggregation phase 2 was dependent on S . aureus viability and an intact bacterial cell wall, and it was mitigated by antibody directed against staphylococcal clumping factor (a fibrinogen-binding protein) and by the cyclooxygenase inhibitor indomethacin . Similarly, aggregation phase 2 was either delayed or absent in three distinct transposon-induced S . aureus mutants with reduced capacities to bind fibrinogen in vitro . In addition, a synthetic pentadecapeptide, corresponding to the staphylococcal binding domain in the C terminus of the fibrinogen delta-chain, blocked aggregation phase 2 . However, phase 2 of aggregation was not inhibited by two synthetic peptides (alone or in combination) analogous to the two principal fibrinogen-binding domains on the platelet glycoprotein (GP) IIb/IIIa integrin receptor: (i) a recognition site on the IIIa molecule for the Arg-Gly-Asp (RGD) sequence of the fibrinogen alpha-chain and (ii) a recognition site on the IIb molecule for a dodecapeptide sequence of the fibrinogen delta-chain . This differs from ADP-induced platelet aggregation, which relies on an intact platelet GP IIb/IIIa receptor with an accessible RGD sequence and dodecapeptide recognition site for fibrinogen . Furthermore, a monoclonal antibody directed against the RGD recognition site on rabbit platelet GP IIb/IIIa receptors failed to inhibit rabbit platelet aggregation by S . aureus . Collectively, these data suggest that S . aureus-induced platelet aggregation requires bacterial binding to fibrinogen but is not principally dependent upon the two major fibrinogen-binding domains on the platelet GP IIb/IIIa integrin receptor, the RGD and dodecapeptide recognition sites.

Infect Immun, 1995 Sep, 63(9), 3373 - 80
Role of the accessory gene regulator (agr) in pathogenesis of staphylococcal osteomyelitis; Gillaspy AF et al.; To examine the role of the accessory gene regulator (agr) in staphylococcal osteomyelitis, we compared a Staphylococcus aureus osteomyelitis isolate (UAMS-1) with a derivative of the same strain (UAMS-4) carrying an inactivated agr locus . Virulence was assessed with a rabbit model of acute, exogenous osteomyelitis . Bacteria were delivered by microinjection into the midradial region of the forelimb . After 4 weeks, UAMS-1 was identified in the bone of 12 of 13 rabbits infected with > or = 2 x 10(6) CFU and 5 of 6 infected with < or = 2 x 10(5) CFU . In contrast, UAMS-4 was found in 6 of 13 infected with the higher dose and 1 of 6 infected with the lower dose . Additionally, on the basis of a five-point scale assessing radiographic evidence of disease, rabbits infected with UAMS-1 had average scores of 2.64 +/- 0.30 (high dose) and 1.43 +/- 0.39 (low dose) while rabbits infected with UAMS-4 had average scores of 0.95 +/- 0.23 (high dose) and 0.63 +/- 0.20 (low dose) . Uninfected controls had an average score of 0.53 +/- 0.08 . The results obtained with UAMS-1 were significantly different from those obtained with UAMS-4 at both doses (P < or = 0.047) . The results obtained with UAMS-4 were not significantly different from those obtained with the controls at either dose of UAMS-4 (P > or = 0.150) . On the basis of a similar five-point scale assessing histopathological evidence of disease, rabbits infected with UAMS-1 had average scores of 2.31 +/- 0.22 (high dose) and 1.96 +/- 0.36 (low dose) while rabbits infected with UAMS-4 had average scores of 1.58 +/- 0.29 (high dose) and 0.83 +/- 0.32 (low dose) . Controls had an average score of 0.33 +/- 0.05 . The results obtained with UAMS-1 were significantly different from those obtained with UAMS-4 at both doses (P < or = 0.040) . However, the results obtained with UAMS-4 were significantly different from the controls only at the high dose of UAMS-4 (P = 0.025) . We conclude that mutation of agr reduces the incidence and severity of disease but does not eliminate the ability to colonize bone and cause histopathological evidence of osteomyelitis.

J Appl Bacteriol, 1995 Sep, 79(3), 347 - 51
Methicillin-resistant Staphylococcus aureus subtyping: interest of combined antibiotyping and esterase electrophoretic typing; Mangeney N et al.; Ninety-four methicillin-resistant Staphylococcus aureus isolates (MRSA) were characterized by means of two typing methods, antibiotyping and esterase electrophoretic typing . Antibiotyping, recorded on the basis of susceptibility testing of 13 antimicrobial agents, allowed the description of 18 antibiotypes, four of which comprised 30, 14, 14 and 12 strains respectively . Esterase electrophoretic typing, based on esterase activity against seven synthetic substrates after polyacrylamide-agarose gel electrophoresis, led to the description of 12 electrophoretic types, two of which were predominant with 60 and 20 strains respectively . The combined use of both typing methods yielded 32 combinations, three of which were predominant with 21, 12 and 11 strains respectively . A good differentiation of strains was achieved, particularly when the antibiotype was correlated to the electrophoretic type . Thus, the combination of antibiotyping with esterase electrophoretic typing may be proposed as a well-suited method for the characterization of MRSA strains.

Klin Lab Diagn, 1995 Sep-Oct, (5), 24 - 6
{Assessment of the effectiveness of noninvasive immunoenzyme method in the detection of Staphylococcus aureus alpha-hemolysin in the washings of nasal mucosa and saliva of healthy individuals}; Musina LT et al.; The efficacies of detection of S . aureus by the bacteriologic method and enzyme immunoassay test system based on F(ab)2 fragments of purified antistaphylococcal antibodies were compared . Washings off the nasal mucosa and salivary samples from 20 normal subjects (medical staff) were examined . The number of findings of S . aureus by the detection of its a-hemotoxin in primary samples was much higher and the time of analysis two times shorter with enzyme immunoassay.

Arch Pediatr, 1995 Sep, 2(9), 883 - 5
{Treatment of atopic dermatitis}; Teillac-Hamel D et al.; There are three components in the treatment of atopic dermatitis: 1) a general treatment (mainly antibiotherapy directed towards Staphylococcus aureus during acute phases), 2) local treatment (local antiseptics, local steroids, emollients), 3) practical advice . In addition, it is of great importance that clear and complete information on the illness should be given to the parents.

Isr J Med Sci, 1995 Sep, 31(9), 558 - 60
Neonatal mastitis--diagnosis and treatment; Efrat M et al.; Neonatal mastitis is an uncommon infection . Twenty-one neonates with mastitis were treated at the Bnai Zion Medical Center and Hillel Yaffe Hospital during the years 1985-92 . Half of them presented with mastitis, and the other half with breast abscess . The most common pathogen was Staphylococcus aureus, which was isolated in 85% of cases . Antibiotic therapy was the initial treatment in all cases except one, and included i.v . orbenin or augmentin . Puncture of six breast abscesses followed the initial antibiotic course, and another five abscesses were treated surgically by incision and drainage . About half the neonates (10 of 21) recovered after antibiotic treatment alone, indicating that aggressive antibiotic therapy is effective in about 50% of cases and, if started immediately upon diagnosis, no additional surgical treatment is necessary . When an abscess was formed, needle aspiration was as effective as incision and drainage.

Eur J Biochem, 1995 Sep 1, 232(2), 658 - 63
Identification of the active-site nucleophile in 6-phospho-beta-galactosidase from Staphylococcus aureus by labelling with synthetic inhibitors; Staedtler P et al.; Kinetic parameters for the inactivation of the 6-phospho-beta-galactosidase of Staphylococcus aureus by a series (fluoro, chloro, bromo) of 2,4-dinitrophenyl-2-deoxy-2-halogeno- galactoside-6-phosphates have been determined . These inhibitors function by the formation of a stabilised glycosyl-enzyme intermediate . Inactivation and reactivation studies indicate that the fluoro derivative is formed most rapidly, but is also hydrolysed fastest . The chloro derivative forms the most stable covalent intermediate . HPLC profiles of V8-protease digestion of native and inhibited protein show significant differences, whereas the inhibited 6-phospho-beta-galactosidase and a point mutant of 6-phospho-beta- galactosidase (E375Q) yield the same proteolytic fragments . The suggestion that E375 is derivatised is strengthened by matrix-assisted laser-desorption ionisation mass spectrometry experiments which show that the two peptides, residues 336-375 and 376-383, are not produced, due to the absence of the expected cleavage at residues 375 and 376 . The reason for the altered proteolysis pattern of the inhibited protein is blocking of the respective V8 cleavage site due to the chemical reaction of the inhibitor at position 375 . Specific modification of the glycosyl bond between the inhibitor and E375 by aminolysis with benzylamine generated a glutamatic-acid-5-benzylamide complex at that position in the peptide . The Edman derivative of the modified E375 appears to be stable and was isolated by Edman degradation of trypsin-digested V8-peptide . It was shown to be identical to an authentic, synthetic sample . From this, it is evident that E375 is the active-site nucleophile of 6-phospho-galactosidase, consistent with previous findings for enzymes in this family.

Br J Urol, 1995 Sep, 76(3), 366 - 70
Immunological infertility among Nigerian men: incidence of circulating antisperm auto-antibodies and some clinical observations: a preliminary report; Ekwere PD; OBJECTIVES: To (i) establish the incidence of circulating antisperm auto-antibodies among infertile men; (ii) relate this incidence to the high prevalence of sexually-transmitted diseases (STDs) in sub-Saharan Africa and; (iii) elucidate the effect of steroid and other therapy on semen quality and subsequent fertility of the patients . PATIENTS AND METHODS: Serum samples from 50 infertile men and 50 age-matched controls were assayed by two agglutination techniques for anti-sperm antibodies . Mean sperm concentrations were determined before and after steroid treatment of patients having antibody titres of 1:64 or above . Serum levels of follicle-stimulating hormone, luteinizing hormone, testosterone and prolactin were also determined by radioimmunoassay in 38 patients . Seminal fluid analysis and culture were performed in 35 patients and testicular histology determined in 21 . RESULTS: Agglutination was demonstrated in 22 of 50 sera (44%), whilst non-agglutinating cytotoxic antibodies were detected in two . Only two of the 50 control sera (4%) were positive . After steroid therapy, antibody titres were significantly decreased and there was a sixfold improvement in mean sperm concentration and a threefold improvement in motility and morphological characteristics . Bacterial (46%) and non-bacterial (17%) infection were recorded in 22 of 35 patients, 13 of whom showed the presence of antisperm antibodies in their sera . Staphylococcus aureus was the commonest single bacterial isolate . Overall, 13 of 29 patients (45%) improved, nine accounting for 12 pregnancies . Pregnancies and/or improvements in semen quality were observed only among patients with mild histological changes . Low testosterone and prolactinaemia occurred in 29% and 21% of the patients, respectively . Among these, anti-sperm antibodies were also recorded in 18% and 13%, respectively . CONCLUSION: The incidence of antisperm antibodies among infertile men is high in Nigeria and may be related to high prevalence of STDs . Immunologically infertile men can be treated successfully with steroids . Concomitant antibiotic and hormone therapy may also be essential in appropriate cases . Clinicians are advised to adopt a multimodal approach to the treatment of male infertility in sub-Saharan Africa . The presence of non-agglutinating cytotoxic antibodies calls for further investigation of the role of complement in the pathogenesis of immunological infertility.

Hepatology, 1995 Sep, 22(3), 687 - 99
In vitro activation of woodchuck lymphocytes measured by radiopurine incorporation and interleukin-2 production: implications for modeling immunity and therapy in hepatitis B virus infection; Cote PJ et al.; Cellular immune responses to hepatitis B virus (HBV) play an important role in the resolution of acute infection . They also influence the course of chronic infection and disease but are inadequate to completely clear the infection . Woodchuck hepatitis virus (WHV) infection of the woodchuck can provide a model to study these processes . Lymphocyte responses of woodchucks were assessed by in vitro proliferation and/or interleukin (IL)-2 assays using mitogen (Concanavalin A {ConA}), cytokine (IL-2), superantigen (Staphylococcus aureus enterotoxin B {SEB}), major histocompatibility complex (MHC) allo-antigen (mixed lymphocyte reaction {MLR}), and viral antigens (woodchuck hepatitis virus core antigen {WHcAg} and woodchuck hepatitis virus surface antigen {WHsAg}) . ConA-stimulated woodchuck lymphocytes underwent cell division based on cell counting experiments and produced IL-2 as detected using an IL-2-dependent murine cell line but failed to incorporate sufficient tritiated thymidine; however, they did incorporate sufficient tritiated adenosine and deoxyadenosine to permit development of a meaningful proliferation assay . The IL-2 assay was sensitive and specific for detection of woodchuck IL-2 induced by mitogen, superantigen, and MLR, as shown by quantitative titration analysis and anti-body neutralization of ConA-supernatant activity . Cyclosporin A and FK506 specifically inhibited ConA- and SEB-induced IL-2 production by woodchuck lymphocytes . Positive two-way MLRs were detected by IL-2 production and proliferation assay between woodchucks from different geographic regions, thus indicating divergence among MHC molecules; however, occasional negative MLR reactions among indigenous pairs of woodchucks indicated that some woodchucks were mutually immunocompatible to some degree . The radioadenosine proliferation assay was sensitive for detecting peripheral blood lymphocyte responses to WHcAg and WHsAg in adult woodchucks with recently resolved acute infections . The above systems should facilitate the design of adoptive therapy and liver transplantation experiments in the woodchuck, and also enable modeling of immune responses that promote and maintain chronic hepadnavirus infection.

East Afr Med J, 1995 Sep, 72(9), 609 - 10
Staphylococcal vertebral osteomyelitis: case report; Longe AC et al.; A 65-year old male Saudi patient presented with rapidly progressive quadriparesis . Lower cervical myelopathy was associated with radiological features of bone destruction, inflammatory disease of the spine and a paravertebral mass . Although tuberculosis and brucellosis are more commonly responsible for this clinical picture in our practice, Staphylococcus aureus was isolated from tissue recovered at surgery . He has been followed up for 12 months and has made an almost complete recovery after surgical decompression of spinal cord and a 2-month course of intravenous flucloxacillin . This case underscores the need for tissue diagnosis in patients presenting with inflammatory paravertebral swellings even in areas endemic for tuberculosis and brucellosis.

J Clin Microbiol, 1995 Sep, 33(9), 2508 - 10
Catheter-related bacteremia associated with coagulase-positive Staphylococcus intermedius; Vandenesch F et al.; We report a case of catheter-related bacteremia in a 63-year-old patient caused by Staphylococcus intermedius . Clinical resolution of the infection was obtained after removal of the intravenous device and antibiotic treatment . This observation emphasizes the risk of confusion between S . intermedius and Staphylococcus aureus if only a coagulase test is done.

J Clin Microbiol, 1995 Sep, 33(9), 2400 - 4
Geographic spread of epidemic multiresistant Staphylococcus aureus clone in Brazil; Teixeira LA et al.; Staphylococcus aureus isolates from five large teaching hospitals and one medium-size community hospital located in geographically distant parts of Brazil, in the south and southeast (Rio de Janeiro, Niteroi, Sao Paulo, Porto Alegre) and in the north (Manaus), were tested for their antibiotic resistance patterns and genetic backgrounds . Eighty-five of the 152 isolates were identified as methicillin-resistant S . aureus (MRSA) by using a combination of an agar dilution screen and a mecA gene-specific DNA probe . All MRSA isolates were resistant to penicillin, erythromycin, gentamicin, oxacillin, and cephalothin, and the majority of isolates (74%) were also resistant to chloramphenicol, sulfamethoxazole-trimethoprim, ciprofloxacin, and clindamycin as well and were susceptible only to vancomycin . Isolates obtained from hospitals in Sao Paulo, Rio de Janeiro, Niteroi, and Porto Alegre (1,600 km from one another) and Manaus (3,700 km from Rio de Janeiro) were examined by a variety of molecular fingerprinting techniques: the nature of the mecA polymorph and Tn554 attachment sites and restriction fragment length polymorphism of genomic DNAs after SmaI restriction and separation of the digested DNA by pulsed-field gel electrophoresis . The overwhelming majority of the isolates shared a common pulsed-field gel electrophoresis pattern and carried mecA polymorph III in combination with Tn554 pattern B, indicating the presence of a single, epidemic MRSA clone spread over large geographic distances of Brazil.

J Clin Microbiol, 1995 Sep, 33(9), 2395 - 9
Evaluation of Rapid ATB Staph for 5-hour antimicrobial susceptibility testing of Staphylococcus aureus . Groupement pour le Dépistage, L'Etude et la Prévention des Infections Hospitalières-Groep ter Opsporing, Studie en Preventie van Infecties in de Ziekenhuizen; Struelens MJ et al.; The accuracy of Rapid ATB Staph (bioMerieux, La Balme-Les Grottes, France) for detection of oxacillin resistance and for detection susceptibility to 11 other antimicrobial agents in 553 and 519 Staphylococcus aureus isolates, respectively, was evaluated by comparing results with those produced by oxacillin agar screen and agar dilution methods, respectively . Further characterization of isolates with discrepant results for oxacillin testing was done by PCR detection of the nuc and mecA genes . By oxacillin agar screening, there were 307 oxacillin-resistant and 246 oxacillin-susceptible isolates . Rapid ATB results were obtained in 5 h for 515 (93.2%) of the isolates tested . Rapid ATB showed 97.0% sensitivity for detection of oxacillin resistance, confirmed by the presence of the mecA gene . After repeat testing of isolates flagged by the ATB software as possible errors, sensitivity increased to 99% for oxacillin-resistant isolates . Essential agreement with agar dilution testing for susceptibility to amoxicillin-clavulanic acid, gentamicin, erythromycin, clindamycin, and ciprofloxacin, as estimated by Youden's J statistic, was > 0.90 . Subpopulations of isolates with significantly increased MICs of amikacin, rifampin, and minocycline, indicating borderline susceptibility, were detected by Rapid ATB and categorized as resistant . Rapid ATB Staph showed adequate accuracy for detection within 5 h of the oxacillin- and multiple-drug-resistant S . aureus isolates currently prevalent in Belgium.

J Clin Microbiol, 1995 Sep, 33(9), 2347 - 52
Incidence of the highly conserved fib gene and expression of the fibrinogen-binding (Fib) protein among clinical isolates of Staphylococcus aureus; Boden Wastfelt MK et al.; We have recently described a 19-kDa fibrinogen-binding protein, termed Fib, which is secreted into the extracellular medium by Staphylococcus aureus . In this study, the presence of the Fib protein and the fib gene among clinical isolates of S . aureus and among other staphylococcal species known to interact with fibrinogen was investigated . This task was pursued at the DNA, mRNA, and protein levels . It was found that the fib gene was unique to S . aureus and highly conserved at the nucleotide level . The Fib protein was produced by all S . aureus strains investigated but was not detected in all bovine mastitis strains, because of proteolytic degradation by simultaneously produced staphylococcal proteases . It was concluded that the fib gene was unique to S . aureus and that it could be used in the identification of S . aureus.

Cathet Cardiovasc Diagn, 1995 Sep, 36(1), 5 - 9; discussion 10
Bacteremia associated with percutaneous transluminal coronary angioplasty; Shea KW et al.; Bacteremia after diagnostic cardiac catheterization is uncommon, but bacteremia after percutaneous transluminal coronary angioplasty (PTCA) has not been studied prospectively . Unlike diagnostic cardiac catheterization, PTCA involves the use of an indwelling arterial sheath after completion of the procedure, which is connected to a pressurized heparin solution, both of which increase the risk of local infection and/or bacteremia . During a 16-week period, we prospectively evaluated patients undergoing 164 PTCA procedures in order to determine the frequency of bacteremia and the significance of fever in this patient population . Blood cultures were obtained from the femoral catheter at the conclusion of the procedure and again 30 min later from the indwelling arterial sheath . Temperature was recorded every 30 min for 2 h following PTCA, then every 4 h over the subsequent 36-hr period . Bacterial isolates were recovered from 23/286 blood cultures (8.0%), with Staphylococcus epidermidis the most common organism present (74%) . Only one isolate of Staphylococcus aureus was considered to represent true bacteremia and corresponded with the only documented infectious complication . Fever, defined as > or = 101 degrees F developed in four (2.4%) patients but was procedure related in only one case . The use of the ipsilateral femoral artery for repeat procedures was not associated with either positive blood cultures or difference in maximum temperature elevation . We conclude the overall risk of bacteremia after PTCA is low; therefore, antimicrobial prophylaxis is not warranted.

Epidemiol Mikrobiol Imunol, 1995 Sep, 44(3), 130 - 8
{Staphylococcus aureus and the human immune system}; Slobodnikova L; The author characterizes factors of virulence of Staphylococcus aureus and its different interactions with immune systems of the host . She describes the immune mechanisms which play a key role in the elimination of staphylococcal infection and where inadequate function leads to the development of staphylococcal disease . The author draws attention to the complicated interrelationship of the complex of different virulence factors of staphylococcal strains involved in inhibition and dysregulation or in stimulation of effector mechanisms of immunity.

APMIS, 1995 Sep, 103(9), 635 - 44
The effect of zinc on bacterial phagocytosis, killing and cytoprotection in human polymorphonuclear leucocytes; Sunzel B et al.; An in vitro study examining the effects of zinc treatment on human PMN cell phagocytosis and killing of Staphylococcus aureus and Staphylococcus epidermidis and the cytoprotection of zinc against staphylococcal toxins . Phagocytosis was studied by transmission electron microscopy using different microbiological techniques, one of which was designed to follow the kinetics of bacterial killing . No effect was found on phagocytosis and bacterial killing . The cytotoxic effects of a crude toxin and an alpha-toxin extracted from Staphylococcus aureus preparations were studied on human PMN cells using the standard 51Cr release assay . Both toxins induced a dose-dependent leakage of 51Cr, indicating cell membrane damage . These results were confirmed by electron microscopy during the phagocytosis of S . aureus, where severe PMN cellular degeneration was observed . The addition of zinc to PMN cells strongly inhibited the release of 51Cr . In conclusion, our results show that zinc in higher than physiological concentrations does not inhibit PMN cell functions such as phagocytosis and intracellular killing of S . aureus and S . epidermidis . The addition of zinc may be beneficial in certain clinical situations, such as wound healing, zinc deficiency and infections involving toxin-producing bacteria, e.g . S . aureus.

Mol Gen Genet, 1995 Aug 30, 248(4), 446 - 58
The agr P2 operon: an autocatalytic sensory transduction system in Staphylococcus aureus; Novick RP et al.; The synthesis of virulence factors and other exoproteins in Staphylococcus aureus is controlled by the global regulator, agr . Expression of secreted proteins is up-regulated in the postexponential growth phase, whereas expression of surface proteins is down-regulated by agr . The agr locus consists of two divergent operons, transcribed from neighboring but non-overlapping promoters, P2 and P3 . The P2 operon sequence, reported here, contains 4 open reading frames, agrA, C, D, and B, of which A and C appear to encode proteins of a classical 2-component signal transduction pathway . The P3 operon specifies a 0.5 kb transcript, RNA III, which is the actual effector of the agr response, and, incidentally, encodes the agr-regulated peptide delta-hemolysin . Transcriptional fusions have shown that both P2 and P3 are agr sensitive (function in an agr+ but not in an agr- background) and deletion analysis has shown that all four of the P2 ORFs are involved; agrA and agrC seem to be absolutely required for the transcriptional activation of the agr locus, whereas agrB and agrD seem to be partially required . Since transcription of P2 requires P2 operon products, the P2 operon is autocatalytic, and is thus admirably suited to the need for rapid production of exoproteins at a time when overall growth is coming to a halt.

Gene, 1995 Aug 30, 162(1), 63 - 8
Sites for co-integration of large staphylococcal plasmids; Sohail M et al.; Site-specific recombination is thought to play an important role in the evolution of multi-resistant plasmids in bacteria, including the human pathogen Staphylococcus aureus (Sa) . A mechanism for site- and orientation-specific recombination between large Sa plasmids was identified in Sa strain 1054 . A replication-thermosensitive derivative of plasmid pI9789::Tn552 (called pS1) was found to form stable co-integrates with the large plasmid pOX1054 in the Sa strain 1054 . Two closely related recombination sites were identified on these plasmids at which recombination occurred to form co-integrates . The sites (rs9789 from plasmid pI9789::Tn552 and rs1054 from pOX1054) were cloned and studies with them showed that the recombination at these sites occurs by a new method . The site rs1054 (27 bp) is deleted and rs9789 (26 bp) is duplicated during recombination . The data show that plasmid pS1 contributes the site for recombination and that the gene(s) encoding the protein(s) involved in recombination are encoded on either pOX1054 or the 1054 chromosome.

Proc Natl Acad Sci U S A, 1995 Aug 29, 92(18), 8313 - 7
Structural features of the invariant chain fragment CLIP controlling rapid release from HLA-DR molecules and inhibition of peptide binding; Kropshofer H et al.; The invariant chain (Ii) prevents binding of ligands to major histocompatibility complex (MHC) class II molecules in the endoplasmic reticulum and during intracellular transport . Stepwise removal of the Ii in a trans-Golgi compartment renders MHC class II molecules accessible for peptide loading, with CLIP (class II-associated Ii peptides) as the final fragment to be released . Here we show that CLIP can be subdivided into distinct functional regions . The C-terminal segment (residues 92-105) of the CLIP-(81-105) fragment mediates inhibition of self- and antigenic peptide binding to HLA-DR2 molecules . In contrast, the N-terminal segment CLIP-(81-98) binds to the Staphylococcus aureus enterotoxin B contact site outside the peptide-binding groove on the alpha 1 domain and does not interfere with peptide binding . Its functional significance appears to lie in the contribution to CLIP removal: the dissociation of CLIP-(81-105) is characterized by a fast off-rate, which is accelerated at endosomal pH, whereas in the absence of the N-terminal CLIP-(81-91), the off-rate of C-terminal CLIP-(92-105) is slow and remains unaltered at low pH . Mechanistically, the N-terminal segment of CLIP seems to prevent tight interactions of CLIP side chains with specificity pockets in the peptide-binding groove that normally occurs during maturation of long-lived class II-peptide complexes.

Schweiz Med Wochenschr, 1995 Aug 26, 125(34), 1592 - 6
{Reconstructive surgery of the mitral valve in the acute stage of bacterial endocarditis . Apropos of 2 cases}; Kalangos A et al.; Two patients in our institution underwent mitral valve reconstruction during the acute phase of Staphylococcus aureus mitral valve endocarditis . In neither case was a pre-existing valve lesion found . Echocardiographic examination revealed severe mitral insufficiency and the extent of valvular lesions . In the first patient, prolapse of the posterior commissure and paracommissural areas was due to ruptured chordae tendinae . In the second patient a perforated abscess was surrounded by vegetations in the median portion of the anterior leaflet and paramedian anterior chordae tendinae were ruptured . The surgical indication was hemodynamic, combined with suspicion of repeated emboli in one case . After a 10-day course of antibiotic therapy, both patients underwent surgical repair by Carpentier's mitral valvuloplasty . During more than 6 months' follow-up no recurrence of endocarditis was observed . Both patients were in class I of the NYHA without echocardiographic evidence of residual mitral regurgitation or stenosis . Early intervention during the acute phase of endocarditis, when mitral valve destruction is not too extensive, allows mitral valvuloplasty which preserves the native valve, eradicates infected tissues and may reduce postoperative mortality and morbidity.

Experientia, 1995 Aug 16, 51(8), 775 - 9
Presence of hsp65 in bacterial extracts (OM-89): a possible mediator of orally-induced tolerance?
Polla BS, Baladi S, Fuller K, Rook G.
Heat shock proteins (HSP) have been implicated in rodent models of autoimmunity, particularly arthritis, and there is suggestive though inconclusive evidence that they may also play a role in human autoimmune disease . The simplest hypothesis is based on molecular mimicry due to the amino-acid sequence homology between mammalian and microbial HSP . Recently OM-89, an extract of several strains of Escherichia coli, has shown some efficacy in the treatment of rheumatoid arthritis (RA) when taken orally . Using species-specific antibodies, we show here that OM-89 contains the 65 kDa HSP (hsp65), while hsp65 was not detected in another bacterial extract containing other microorganisms, including Staphylococcus aureus (OM-85) . We suggest that if the human homologue of hsp65 is a relevant target antigen in the human disease, the efficacy of the preparation could be due to induction of oral tolerance or to switching the Th1 response towards Th2 . Alternatively, even if the human hsp65 is not a target molecule in RA joints, OM-89 may evoke bystander suppression of joint inflammation via induction of TGF beta-secreting effector cells . These hypotheses should be tested in further studies.

J Immunol, 1995 Aug 15, 155(4), 2067 - 76
Mice with the xid B cell defect are less susceptible to developing Staphylococcus aureus-induced arthritis; Zhao YX et al.; To investigate the role of B cells in the development of experimental Staphylococcus aureus-induced arthritis, we used X-linked immunodeficiency (xid) mice that carry a Bruton's tyrosine kinase mutation affecting the function of B cells . NFR/N.xid and congenic NFR/N mice were inoculated i.v . with a toxic syndrome toxin-1 producing S . aureus LS-1 strain . B cell-deficient NFR/N.xid mice developed less frequent (p < 0.01) and less severe (p < 0.01) arthritis than NFR/N mice did . These clinical findings were corroborated by histopathologic evaluation, indicating that NFR/N.xid mice had significantly lower (p < 0.01) erosivity of the disease . Interestingly, infected NFR/N.xid mice showed decreased bacterial burden in blood, joints, and other organs compared with the control mice . Serologic studies displayed poor B cell responses to staphylococcal cell walls, toxic shock syndrome toxin-1, and ssDNA, accompanied by a low level of Igs in infected NFR/N.xid mice . More importantly, xid defect affected cytokine profile . The in vitro experiments showed that the lymphocytes from NFR/N.xid mice had low IL-6, but high IFN-gamma production upon stimulation with staphylococcal cell walls compared with NFR/N mice . Furthermore, the in situ hybridization technique revealed the relative increase of IFN-gamma, but marked decrease of IL-1 beta mRNA expression in spleens of infected NFR/N.xid mice . No significant difference in IL-4, IL-10, and TNF-alpha mRNA expression was found between both strains . Our findings demonstrate that B cells may, directly or indirectly, contribute to the pathogenesis of septic arthritis . The results indicate that increased IFN-gamma production along with low IL-6 and IL-1 beta synthesis found in xid mice may provide a more favorable outcome of S . aureus arthritis.

Structure, 1995 Aug 15, 3(8), 769 - 79
Crystal structure of the superantigen enterotoxin C2 from Staphylococcus aureus reveals a zinc-binding site; Papageorgiou AC et al.; BACKGROUND: Staphylococcus aureus enterotoxin C2 (SEC2) belongs to a family of proteins, termed 'superantigens', that form complexes with class II MHC molecules enabling them to activate a substantial number of T cells . Although superantigens seem to act by a common mechanism, they vary in many of their specific interactions and biological properties . Comparison of the structure of SEC2 with those of two other superantigens--staphylococcal enterotoxin B (SEB) and toxic shock syndrome toxin-1 (TSST-1)--may provide insight into their mode of action . RESULTS: The crystal structure of SEC2 has been determined at 2.0 A resolution . The overall topology of the molecule resembles that of SEB and TSST-1, and the regions corresponding to the MHC class II and T-cell receptor binding sites on SEB are quite similar in SEC2 . A unique feature of SEC2 is the presence of a zinc ion located in a solvent-exposed region at the interface between the two domains of the molecule . The zinc ion is coordinated to Asp83, His118, His122 and Asp9* (from the neighbouring molecule in the crystal lattice) . Atomic absorption spectrometry demonstrates that zinc is also bound to SEC2 in solution . CONCLUSIONS: SEC2 appears to be capable of binding to MHC class II molecules in much the same manner as SEB . However, structure-function studies have suggested an alternative binding mode that involves a different site on the toxin . The zinc ion of SEC2 lies within this region and thus may be important for complex formation, for example by acting as a bridge between the two molecules . Other possible roles for the metal cation, including a catalytic one, are also considered.

Orv Hetil, 1995 Aug 13, 136(33), 1769 - 75
{Spinal epidural abscess}; Vidovszky T; The aspecific spinal epidural abscess is an uncommon cause of the spinal cord compression . Forty eight patients presented with epidural abscess were operated on during the last 37-year-period in the National Institute of Neurosurgery Budapest . Experiences with the diagnostic methods and the results of the treatment of these patients are analysed . Based on the case history data, preoperative symptoms and operative findings 31 male and 17 female patients (their age ranged from 15-64 years) have been selected in acute, subacute and chronic groups . Localized backpain, acceleration of the blood sedimentation, leukocytosis and fever were significant findings which were followed by sings of radicular or medullary compression . This clinical picture developed rapidly in the acute group . Aspecific abscess most commonly appeared in thoracal or lumbal localisation and was rarely found in the ventral area . Staphylococcus aureus was the causative organism in nearly 100 percent of the cases . Surgery carried out before the onset of the neurological deficits according to proper CSF examination and myelography, proved to be favourable . Early diagnosis and emergency operation led to a significant improvement of the outcome.

Schweiz Med Wochenschr, 1995 Aug 8, 125(31-32), 1477 - 82
{Intramuscular injections--an outdated form of administration? 6 cases of Staphylococcus aureus sepsis following intramuscular injections}; Rossi L et al.; Intramuscular injections can lead to local and systemic complications, such as abscess and sepsis . These are often caused by Staphylococcus aureus, occur in immunocompromised as well as in immunocompetent persons, and often need extensive medical and surgical treatment . We describe 6 cases with sepsis and multiple abscesses caused by Staphylococcus aureus after intramuscular injections . In view of possible serious complications, the indication for intramuscular injection as a method of drug administration is critically analyzed.

Contemp Orthop, 1995 Sep, 31(3), 159 - 64
Pyogenic vertebral osteomyelitis: report of a series of 23 patients; Stefanovski N et al.; Vertebral osteomyelitis, an infectious disease with vague manifestations, can be difficult to diagnose . Although vertebral osteomyelitis represents only 2-4% of bone infections, the consequences are often grave and disabling, even fatal, when untreated . A series of 23 cases is reported in which patient records were reviewed to determine the etiology and appropriateness of diagnosis and management . Information on treatment and follow-up after discharge was obtained from outpatient progress notes, records from subsequent hospital admissions, and telephone interviews of patients . Pyogenic vertebral osteomyelitis was diagnosed by positive needle or open biopsy tissue cultures, positive blood cultures in the appropriate clinical setting, or diagnostic histopathology . Staphylococcus aureus was grown from 75% (15/20) of patient cultures, Escherichia coli from 15%, and Staphylococcus epidermidis from 10% . Overall, 87% (20/23) of these patients were disease-free at follow-up . The experience with this series of patients demonstrates that early diagnosis aided by MRI ensures a high cure rate and low complication rate.

Biochem Biophys Res Commun, 1995 Aug 4, 213(1), 239 - 48
Purification and characterization of human pancreatic polypeptide expressed in E . coli; Griko YV et al.; The region of cDNA encoding human pancreatic polypeptide (hPP) was obtained by polymerase chain reaction (PCR) and subcloned into an expression vector . The pancreatic polypeptide gene was expressed in Escherichia coli in two versions: as a cleavable fusion protein with IgG-binding synthetic ZZ domains of protein A from Staphylococcus aureus or with the 1-48 fragment of lambda Cro repressor . Site-specific hydrolysis by hydroxylamine was used to cleave the fusion protein, releasing the human polypeptide . The structure of the obtained hPP has been studied by scanning microcalorimetry and circular dichroism spectrometry . It has been shown that hPP in solutions close to neutral has a compact and unique spatial structure with an extended hydrophobic core . This structure is stable at 20 degrees C and co-operatively breaks down upon heating from this temperature.

Mol Microbiol, 1995 Aug, 17(4), 769 - 79
Rearrangements in the staphylococcal beta-lactamase-encoding plasmid, pIP1066, including a DNA inversion that generates two alternative transposons; Derbise A et al.; The plasmid plP1066, harboured by by a methicillin-resistant Staphylococcus aureus strain isolated in France, carries genes specifying beta-lactamase . This plasmid undergoes numerous rearrangements . One of these was insertion, between the genes binR and sin encoding resolvases, of a 16 kb element which displayed the characteristic features of a transposon . This putative transposon, named Tn5404, carried genes encoding proteins involved in its transposition, as well as a resolution system, which were indistinguishable from those of the S . aureus transposon Tn552 . These were: p480 encoding a probable transposase, p271 encoding a putative ATP-binding protein, binL encoding a resolvase, and a resolution site, resL . In addition, Tn5404 carried aminoglycoside-resistance genes (aphA, str) and the insertion sequence IS1181 . Tn5404 contained at its termini 116 bp imperfect inverted repeats, similar to those of Tn552, and was flanked by 6 bp direct repeats . Insertion of Tn5404 close to resR and to the structural and regulatory beta-lactamase genes (blaZ, blal, blaR1) of pIP1066, generated a 3.5 kb invertible segment flanked by inversely repeated resolution sites (resR, resL) . This invertible segment, which carried p480, p271 and binL, generated in Tn552 or Tn5404, depending on its orientation . Thus, these two transposons share their transposition and resolution systems.

Antibiot Khimioter, 1995 Aug, 40(8), 23 - 7
{Antibiotic sensitivity of hospital strains of Staphylococcus}; Shubitidze AE et al.; Seven hundred and six clinical strains of Staphylococcus spp . isolated in different cities of the former USSR in 1986-1992 were investigated . It was shown that the isolates had multiple drug resistance . The hospital strains of Staphylococcus aureus more frequently contained the plasmids with the molecular weight of 21.0 and 18.0 MD as well as the plasmids with the molecular weight of 1.4 MD determining erythromycin resistance . The plasmids with the molecular weight of 3.0 and 2.6 MD were detected which determined streptomycin and tetracycline resistance respectively . Lincomycin and carbenicillin resistance was determined by the chromosomal genes . The elimination studies demonstrated that gentamicin resistance could be determined by the extrachromosomal genes.

Acta Ophthalmol Scand, 1995 Aug, 73(4), 355 - 7
Ocular side effects associated with 13-cis-retinoic acid therapy for acne vulgaris: clinical features, alterations of tearfilm and conjunctival flora; Egger SF et al.; Isotretinoin (13-cis-retinoic acid) is commonly used for the treatment of acne vulgaris . We included 55 patients in this prospective study, and inspected them before, while and after therapy with isotretinoin regarding ocular side effects . Careful slit-lamp inspection, measurement of break-up-time and Schirmer-test and microbiological investigations of the conjunctival flora were performed . While staphylococcus aureus was cultured from the conjunctival sac before application of isotretinoin in 7.3%, this percentage increased to 61.8% during therapy . A pathological decrease of break-up-time was realized in 69.1% of the cases, the development of blepharitis in 40% . But in spite of the alteration of conjunctival flora, bacterial conjunctivitis developed in just 7.3% of the cases . However, only 34.5% of the patients showed symptoms of a conjunctivitis sicca, in spite of the impressive diminution of break-up-time in so many cases . All ocular side effects of isotretinoin were treatable and disappeared completely within 1 month after stopping therapy.

Artif Organs, 1995 Aug, 19(8), 801 - 7
Hemodialysis with cellulose membranes primes the neutrophil oxidative burst; Ward RA et al.; Hemodialysis with cellulose membranes causes a complement-mediated neutropenia . Changes in neutrophil function have also been reported; however, it is unclear if these changes indicate a direct effect of the membrane on neutrophils or if they are a consequence of the neutropenia . We tested the hypothesis that neutrophil oxidative burst activity is enhanced during dialysis with cellulose membranes . Resting and Staphylococcus aureus-stimulated H2O2 production were determined predialysis and in blood entering and leaving the dialyzer during the first 30 min of dialysis and in blood leaving the membrane module in a single-pass on-line model of hemodialysis . Resting H2O2 production increased slightly but significantly during the first 30 min of dialysis . Transit of neutrophils through the dialyzer caused a marked increase in stimulated H2O2 production, indicating priming of the oxidative burst . However, priming was limited to the first 5 min of dialysis before the onset of neutropenia . In contrast, stimulation and priming of H2O2 production persisted throughout 30 min of single-pass on-line perfusion . These results indicate that cellulose membranes both stimulate and prime neutrophil oxidative burst activity but that these effects are partially obscured by neutropenia.

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