Rinsho Byori, 1994 Dec, 42(12), 1227 - 33 {Molecular typing of the methicillin resistance determinant (mec) of clinical strains of Staphylococcus based on mec hypervariable region length polymorphisms}; Nishi J et al.; We used a method for molecular typing of the methicillin resistance determinant (mec) based on the size of the mec-associated hypervariable region amplified by the polymerase chain reaction (PCR) to examine 106 methicillin-resistant Staphylococcus aureus (MRSA), 9 methicillin-resistant (Mcr) S . epidermidis and 5 Mcr S . haemolyticus clinical isolates . In the 106 MRSA isolates, 5 sizes of PCR products were observed . The MRSA isolates were grouped into five hypervariable region (HVR) genotypes on the basis of the size of the PCR product . The PCR products amplified from 8 of 9 Mcr S . epidermidis isolates were the same as products amplified from MRSA isolates, which was confirmed by the PCR-SSCP (single-strand conformation polymorphism) method . In 32 methicillin-susceptible isolates, the target gene was not amplified . This method is thought to be useful in epidemiological investigations of nosocomial infections caused by MRSA . This is the first typing method capable of comparing the mec determinants of MRSA isolates and Mcr CNS isolates to establish the origin of the mec determinant. Yakugaku Zasshi, 1994 Dec, 114(12), 1015 - 20 {A quantitative determination of inducibility by 14 membered ring macrolide antibiotics in inducible resistant Staphylococcus aureus to macrolide-lincosamide-streptogramin B antibiotics}; Kobayashi H et al.; Using Staphylococcus aureus ISP447 strain, which shows inducible resistance to macrolide-lincosamide-streptogramin B (MLS) antibiotics, the extent of MLS-resistance induced by several macrolide antibiotics {erythromycin (EM), oleandomycin (OL), or roxithromycin (RXM)} was determined in terms of a relative ratio of a growth rate of the induced cells in the presence of a challenging drug, rokitamycin (RKM), to that of uninduced cells in the absence of RKM . The ratio was referred to as a relative inducibility (%) . The inducibility was obtained at an optimum-induced condition by considering the following factors: (1) exponentially growing cells, (2) the optimum concentration of an inducer drug, i.e., 50, 150, and 150 ng/ml for EM, OL, and RXM, respectively, (3) a 3-h previous incubation at 37 degrees C in the presence of the inducer, and (4) 300 ng of RKM/ml, which is found to be optimum for induced cells to challenge, because of having no inducer activity . Using these qualification methods, inducibilities of EM, OL, and RXM as an inducer were 100.4, 27.9 and 81.1%, respectively . This method is allowed to be useful for the analysis of a structure-inducibility relationship. Ophthal Plast Reconstr Surg, 1994 Dec, 10(4), 267 - 70 Hydroxyapatite orbital implant abscess: histopathologic correlation of an infected implant following evisceration; Ainbinder DJ et al.; A 60-year-old diabetic man with a history of ocular trauma and absolute glaucoma underwent evisceration with placement of an 18 mm hydroxyapatite orbital implant . The host scleral shell was left intact with no posterior opening for vascular ingrowth . One year later the patient presented with implant exposure, limited fibrovascular ingrowth into the implant, and a Staphylococcus aureus orbital abscess . The implant was removed, and pathology demonstrated suppurative inflammation with limited vascular ingrowth . This patient's risk factors for an implant-associated wound infection included diabetes, impaired wound healing, history of trauma, early implant exposure, and delayed fibrovascular ingrowth . Awareness of the infectious complications of any orbital implant including hydroxyapatite allows the surgeon to alter management strategies in an attempt to reduce such risk . Surgeons may consider posterior scleral portals with evisceration surgery to facilitate hydroxyapatite vascularization . Patient selection, implant size, and surgical technique are key factors for a management strategy designed to reduce the risk of implant infection. Eur J Immunol, 1994 Dec, 24(12), 3237 - 40 Interleukin-8 differentially modulates interleukin-4- and interleukin-2-induced human B cell growth; Kimata H et al.; The effect of interleukin-8 (IL)-8 on human B cell growth, as determined by thymidine uptake and viable cell numbers was studied . IL-8 inhibited IL-4-induced growth of B cells costimulated with anti-mu antibodies (Ab) or Staphylococcus aureus Cowan strain I (SAC) in a dose-dependent fashion . In contrast, IL-8 did not inhibit IL-2-induced growth of B cells . The IL-8-mediated inhibition was specific, since it was blocked by anti-IL-8 mAb but not by control IgG1 . Moreover, anti-tumor necrosis factor-alpha (anti-TNF-alpha) Ab blocked IL-8-mediated inhibition . On the other hand, TNF-alpha, but not other cytokines including IL-1 beta, IL-3, IL-5, IL-6, interferon-alpha (IFN-alpha) or IFN-gamma, inhibited IL-4-mediated growth, and inhibition by TNF-alpha was blocked by anti-TNF-alpha Ab but not by control IgG . IL-4 had no effect on TNF-alpha binding by B cells while it decreased TNF-alpha production by B cells . IL-8 had no effect in binding of IL-4, IL-2 or TNF-alpha by B cells, however, it enhanced TNF-alpha production by B cells . These results indicate that IL-8 inhibited IL-4-induced human B cell growth by enhancement of endogenous TNF-alpha production. Yan Ke Xue Bao, 1994 Dec, 10(4), 251 - 3 {The clinical observation of China-made ofloxacin eye drops in the treatment of bacterial infection of the external eye}; Lin B et al.; Ninety patients with extraocular infection were observed . After bacteriological examination on all of the patients, we found that staphylococcus epidermidis was the main pathogen and accounted for 43.3% . Next to it were staphylococcus aureus (31.1%), saprophytic staphylococcus was (5.6%), diplococcus catarrhus (3.3%) and moraxella sp . (2.2%) . All the patients were treated with Ofloxacin eye drops produced in Guangzhou . The bacteriological examination showed negative in 98.9% of the patients, which proved this medicine has high antibiotic power. Yan Ke Xue Bao, 1994 Dec, 10(4), 241 - 3, 250 {Bacteriological detection of anterior chamber aspirate after extracapsular cataract extraction with intraocular lens implantation}; Yang W et al.; We cultured the anterior chamber aspirates of 83 patients with extracapsular cataract extraction and IOL implantation . Of the 83 cases, 5 (6.1%) had positive aspirates including 3 cases of staphylococcus epidermidis and 2 cases of staphylococcus aureus . No eye in our study developed bacterial endophthalmitis . Our study suggests that the anterior chamber of human eyes is capable of clearing a low inoculum of bacteria after cataract surgery without the development of bacterial endophthalmitis. Minerva Chir, 1994 Dec, 49(12), 1299 - 303 {The capability of silicone prostheses to promote postoperative infections . An in-vitro study}; Arciola CR et al.; The authors have quantitatively evaluated and compared in vitro the Staphylococcus aureus and Staphylococcus epidermidis strain adhesiveness on various types of silicone used in surgery . The results show that there are some adhesiveness differences between the two assayed strains (S . epidermidis adhesiveness > S . aureus adhesiveness) and among silicone types (adhesiveness on soft silicones > adhesiveness on hard silicones) . However all silicones are suitable substrata for in vitro bacterial adhesion. J Antimicrob Chemother, 1994 Dec, 34(6), 921 - 30 Decreased bactericidal activity during the period of the postantibiotic effect; Gudmundsson S et al.; Standard and clinical strains of Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae were subjected to continuous exposure to beta-lactams and aminoglycosides during the postantibiotic effect phase induced by rifampicin or erythromycin (for S . aureus) . A significant inhibition of bactericidal activity by these agents during the PAE period was observed . The degree of inhibition was dependent both on the class of antimicrobial agent (beta-lactams > aminoglycosides) and the microorganism (Gram-negative bacilli > S . aureus). J Antimicrob Chemother, 1994 Dec, 34(6), 909 - 20 Role of beta-lactamase of methicillin-susceptible Staphylococcus aureus in resistance to first-generation oral cephems both in vitro and in vivo; Takenouchi T et al.; Cefaclor, among the oral cephalosporins tested, showed the largest inoculum effect with respect to MIC values for 61 clinical isolates of methicillin-susceptible Staphylococcus aureus, including 39 beta-lactamase producing strains . These 39 strains were divided into eight type A, 29 type B or C, and two type D producers, by comparisons of specific activities to three substrates . Two producers, one each of types A and C, were further studied to investigate the effect of beta-lactamase on staphylococcal resistance to several beta-lactams . Concentrations of cefaclor and cephalexin in cultures of these strains decreased rapidly, whereas hydrolysis of these drugs by the purified beta-lactamases was moderate to low as detected by spectrophotometric assay . Cefaclor showed high affinities for penicillin-binding proteins 1, 2, and 3 of both beta-lactamase producers and their respective penicillinase-non-producing mutants . In experimental intraperitoneal infections in mice, cefaclor was therapeutically effective against both mutants, showing 50% effective doses of less than 10 mg/kg/dose . In contrast, it was not satisfactory against the parent strains, requiring greater-than-10-fold increases in concentration for the same degree of survival . We concluded that resistance to first-generation oral cephems seen both in vitro and in vivo was due mainly to the beta-lactamase production. J Antimicrob Chemother, 1994 Dec, 34(6), 899 - 907 In-vitro antibacterial activity of fusidic acid alone and in combination with other antibiotics against methicillin-sensitive and -resistant Staphylococcus aureus; Drugeon HB et al.; The activity of fusidic acid alone and in combination with gentamicin, oxacillin, rifampicin, fosfomycin and ciprofloxacin was investigated against 36 strains of Staphylococcus aureus . The fusidic acid MIC50 of 0.06 mg/L and MIC90 of 0.125 mg/L were determined for 36 strains . Bactericidal activity of fusidic acid was confirmed . Several types of interaction were observed with other antibiotics: dominant effect of fusidic acid over gentamicin; synergy with rifampicin for three or four strains; indifference or dominance with oxacillin, indifference with vancomycin, indifference or dominance with fosfomycin, and indifference with ciprofloxacin . Antagonism was seen with only one strain, following exposure to fusidic acid and rifampicin. J Antimicrob Chemother, 1994 Dec, 34(6), 885 - 97 The effect of Triton X-100 on the in-vitro susceptibility of methicillin-resistant Staphylococcus aureus to oxacillin; Komatsuzawa H et al.; The effect of the non-ionic detergent, Triton X-100, on the in-vitro activity of oxacillin against methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) strains of Staphylococcus aureus was investigated . In the presence of Triton X-100, the MICs of oxacillin for both MRSA and MSSA isolates were reduced; this enhancing effect was particularly marked for the MRSA strains . Triton X-100 therefore counteracted the resistance to methicillin encoded by mecA . In the presence of oxacillin at subinhibitory concentrations, Triton X-100 induced the bacteriolysis of MRSA and potentiated the autolysis of these organisms . However, the detergent had no effect on the bacteriolytic enzyme profile or the susceptibility of the bacterial cell wall to bacteriolytic enzymes, nor did it promote the binding of oxacillin to the penicillin-binding protein (PBP) 2A . On the other hand, it stimulated the release from the bacteria of acylated lipoteichoic acid (LTA), a putative endogenous regulator of autolysins . Autolytic enzyme-deficient MRSA mutants were equally as sensitive as the parent strain to the effect of Triton X-100 on susceptibility to oxacillin . These results indicate that the enhanced in-vitro activity of oxacillin against MRSA in the presence of Triton X-100 cannot be accounted for simply by the induction of bacteriolysis following activation of autolytic enzymes by the detergent-stimulated release of LTA. Jpn J Pharmacol, 1994 Dec, 66(4), 451 - 6 Enhancement by ascorbic acid 2-glucoside or repeated additions of ascorbate of mitogen-induced IgM and IgG productions by human peripheral blood lymphocytes; Tanaka M et al.; In this study, the effect of ascorbic acid 2-glucoside (AA-2G), a stable derivative of ascorbic acid (AsA), or repeated additions of ascorbate on antibody productions by human peripheral blood lymphocytes (PBLs) was examined, and the physiological function of AsA was evaluated . When human PBLs were stimulated with Staphylococcus aureus Cowan I or pokeweed mitogen, AA-2G remarkably increased the numbers of IgM- and IgG-secreting cells which were detected by enzyme-linked immunospot assay . Although a single addition of ascorbate was without effect, the effect of AA-2G was remarkably inhibited by the addition of castanospermine, an alpha-glucosidase inhibitor; and moreover, repeated additions of AsA to the culture medium during the culture period enhanced the response to the same level as did a single addition of AA-2G . These results indicate that AsA has the ability to stimulate the immunoglobulin productions by AA-2G . The phytohemagglutinin-induced proliferative response of PBLs was also stimulated by AA-2G . The intracellular AsA content in PBLs cultured with AA-2G was maintained at relatively high levels during the culture period, whereas the content with a single dose of AsA reached nearly zero by the end of the experiment . These in vitro findings suggest that AA-2G and AsA function as potent immunostimulators of antibody production in humans and that the intracellular AsA content is a key parameter for establishing the immune response of PBLs. Protein Eng, 1994 Dec, 7(12), 1463 - 70 The stability and unfolding of an IgG binding protein based upon the B domain of protein A from Staphylococcus aureus probed by tryptophan substitution and fluorescence spectroscopy; Bottomley SP et al.; The stability and unfolding of an immunoglobulin (Ig) G binding protein based upon the B domain of protein A (SpAB) from Staphylococcus aureus were studied by substituting tryptophan residues at strategic locations within each of the three alpha-helical regions (alpha 1-alpha 3) of the domain . The role of the C-terminal helix, alpha 3, was investigated by generating two protein constructs, one corresponding to the complete SpAB, the other lacking a part of alpha 3; the Trp substitutions were made in both one- and two-domain versions of each of these constructs . The fluorescence properties of each of the single-tryptophan mutants were studied in the native state and as a function of guanidine-HCl-mediated unfolding, and their IgG binding activities were determined by a competitive enzyme-linked immunosorbent assay . The free energies of folding and of binding to IgG for each mutant were compared with those for the native domains . The effect of each substitution upon the overall structure and upon the IgG binding interface was modelled by molecular graphics and energy minimization . These studies indicate that (i) alpha 3 contributes to the overall stability of the domain and to the formation of the IgG binding site in alpha 1 and alpha 2, and (ii) alpha 1 unfolds first, followed by alpha 2 and alpha 3 together. J Virol Methods, 1994 Dec, 50(1-3), 29 - 41 Rapid coagglutination test for the detection and typing of foot and mouth disease virus; Montassier HJ et al.; Protein A containing Staphylococcus aureus was used to develop a coagglutination (COA) test for the detection and typing of foot and mouth disease virus (FMDV) O, A and C serotypes in infected cells and tissues . Different batches and amounts of guinea pig anti-FMDV sera were assessed to optimize the preparation of COA conjugates . The sensitivity and specificity of the COA Test for the detection of FMDV O, A and C serotypes and heterologous viruses was also characterized . Comparison between the COA Test and complement fixation test for the detection and typing of FMDV obtained from extracts of tongue epithelial tissues from infected cattle revealed high agreement in the results and indicated a potential application of the COA Test for the direct diagnosis of viruses. Int J Immunopharmacol, 1994 Dec, 16(12), 977 - 84 Immunostimulating effects of protein A in immunosuppressed aflatoxin-intoxicated rats; Raisuddin S et al.; Aflatoxin B1, the potent carcinogenic compound produced by the Aspergillus flavus group of fungi on food and feed, induces immunosuppressive effects in rodents . In this communication, we report an immunomodulatory approach to abrogate aflatoxin B1-induced immunotoxicity in rats using protein A of Staphylococcus aureus Cowan 1 . We have earlier demonstrated that protein A can protect the animals from toxicities induced by a number of drugs, chemicals and toxins . In the present study various combinations of aflatoxin B1 exposure and protein A treatment in animals were used . It was observed that protein A could provide protection to animals from aflatoxin B1-induced immunotoxicity, as measured by a battery of tests assessing cell-mediated immunity (CMI) profile of the host . Various parameters showing suppression of CMI following aflatoxin B1 exposure were reverted back towards normalcy in protein A-treated animals . It is concluded that protein A may prove to be a useful agent to protect the host from aflatoxin immunotoxicity, in view of its stimulatory effects on various immune functions even after their initial depression due to aflatoxin B1. Int J Food Microbiol, 1994 Dec, 24(1-2), 171 - 8 Enterotoxigenic Escherichia coli and Staphylococcus aureus in fish and seafood from the southern region of Brazil; Ayulo AM et al.; An investigation to evaluate the microbiological condition and safety of fish and seafood commonly harvested at the coast of Santa Catarina State and sold in Florianopolis was undertaken . One hundred and seventy-five samples of fish and fish fillets (Cynoscion leiarchus), shrimp tails (Peneaus paulensis), shellfish-meat (Anomalocardia brasiliensis and Metilus edulis), and crab-meat (Callinectes sapidus) were collected from markets and examined within 4 h of purchase . For isolation and enumeration of Escherichia coli the methods used were those of Speck et al . (1975) (Method 1) and Fishbein et al . (1976) (Method 2); for S . aureus, methods recommended by the U.S . Food and Drug Administration were used including biochemical identification of the strains . E . coli was more frequently detected with Method 1 than Method 2 . Of 317 E . coli strains tested for STG and LT II toxins, only one (isolated from shellfish-meat) produced ST and none produced LT II toxin . S . aureus was isolated from 20% of 175 samples examined, including 60% of samples of shellfish-meat . Only nine of 109 S . aureus strains produced enterotoxins, including enterotoxin A (4), D (1) and AB (4) . It is concluded that greater care must be taken to reduce contamination of fish and seafood during harvesting and post-harvest handling. Antimicrob Agents Chemother, 1994 Dec, 38(12), 2810 - 6 Effect of magnesium complexation by fluoroquinolones on their antibacterial properties; Lecomte S et al.; By using infrared and 19F nuclear magnetic resonance spectroscopies, we localized the binding site and measured the affinity of magnesium for six fluoroquinolones . It was proven that magnesium is situated between the ketone and the carboxylate groups . We determined the binding constants for the 1:1 Mg(2+)-drug complex in solution . Sparfloxacin and pefloxacin, with affinity constants (Ka) of (10.1 +/- 0.6) x 10(2) M-1 and (21 +/- 1) x 10(2) M-1, respectively, were the least and the most bound, respectively . The trend of the affinities of the assayed fluoroquinolones for magnesium was correlated with their antimicrobial activities against four bacteria and with their accumulation by these bacteria . The reference strain, Escherichia coli KL16, and two resistant mutants, NalA (gyrase mutation) and NalB (uptake defect), plus Staphylococcus aureus 209P were used . It appeared that, in every case, an impairment of accumulation is responsible for the increase in the MICs observed upon the addition of magnesium. Antimicrob Agents Chemother, 1994 Dec, 38(12), 2702 - 9 Pharmacodynamics of levofloxacin, ofloxacin, and ciprofloxacin, alone and in combination with rifampin, against methicillin-susceptible and -resistant Staphylococcus aureus in an in vitro infection model; Kang SL et al.; The pharmacodynamic properties of levofloxacin (an optically active isomer of ofloxacin), ofloxacin, and ciprofloxacin, alone and in combination with rifampin, were evaluated over 24 to 48 h against clinical isolates of methicillin-susceptible and -resistant Staphylococcus aureus (MSSA 1199 and MRSA 494, respectively) in an in vitro infection model . The incidence of the emergence of resistance among the test strains was also determined . The fluoroquinolones were administered to simulate dosage regimens of 200 mg, 400 mg given intravenously (i.v.) every 12 h (q12h), and 400 and 800 mg given i.v . q24h . Rifampin was dosed at 600 mg i.v . q24h . Although the MICs and MBCs of the quinolones were similar (< or = 0.49 microgram/ml), levofloxacin was the most potent agent in time-kill studies on the basis of the time required to achieve a 99.9% reduction in the number of log10 CFU per milliliter (e.g., with the regimen of levofloxacin {400 mg q24h, 6.5 h} versus ofloxacin {12.5 h}, P < 0.024, and levofloxacin versus ciprofloxacin {6.5 versus 9.0 h}, P < 0.0017) against MSSA 1199 . The killing activity of levofloxacin was similar to that of ofloxacin against MRSA 494 (time to achieve a 99.9% reduction in the number of log10 CFU per milliliter, 11.1 versus 13.8 h, respectively) . Levofloxacin and ofloxacin dosed once daily demonstrated greater bactericidal activity than when they were dosed twice daily against MSSA 1199 . Resistance to levofloxacin or ofloxacin was not observed with any dosage regimen . Furthermore, resistance to ofloxacin was not detected when the half-life was reduced from 6 to 3 h . Regrowth and stable resistance (65-fold increase in the MIC for MSSA 1199; 16-fold increase in the MIC for MRSA 494) were noted within 24 h of exposure to ciprofloxacin at 200 mg q12h . Combination therapy with rifampin prevented the emergence of resistance to ciprofloxacin . Neither DNA gyrase alteration nor an energy-dependent efflux process mediated by the norA gene appeared to be responsible for the resistance observed . Our data suggest that with levofloxacin there is a more rapid onset of bactericidal activity than with ofloxacin or ciprofloxacin against MSSA 1199 and that the activity of levofloxacin is similar to that of ofloxacin but better than that of ciprofloxacin against MRSA 494 . Resistance was noted only after exposure to the low dose of ciprofloxacin . Resistance to ofloxacin did not develop even when the pharmacokinetics of the drug were set to equal those of ciprofloxacin, suggesting that ofloxacin differs from ciprofloxacin irrespective of time of exposure . The resistance to ciprofloxacin that developed in our vitro model may be mediated by the cfx-ofx locus, which has been shown to be associated with low-level fluoroquinolone resistance . Overall, levofloxacin demonstrated potent bactericidal activity against S . aureus, without the emergence of resistance in our infection model . Quinolones dosed once daily were more effective than equivalent dosages administered twice daily . The addition of rifampin was not synergistic but prevented the emergence of ciprofloxacin resistance. Am J Infect Control, 1994 Dec, 22(6), 340 - 5 Control of methicillin-resistant Staphylococcus aureus in a pediatric burn unit; Sheridan RL et al.; BACKGROUND: Control of methicillin-resistant Staphylococcus aureus (MRSA) is particularly difficult in burn units, which are often cited as sources of hospital-wide MRSA outbreaks . We developed a successful MRSA control program and document here its apparent effectiveness in controlling MRSA transmission in a pediatric burn unit . METHODS: An MRSA control program that included surveillance culturing, clinician feedback, flexible, site-specific isolation, and a list of known carriers was consistently applied in a pediatric burn unit through a 7-year period . Microbiology reports of MRSA isolates from patients and environmental surfaces and records of all patients from whom MRSA was isolated were reviewed . RESULTS: During calendar years 1985 through 1991, a total of 991 acutely burned children were admitted to the Boston unit of the Shriners Burns Institute . Forty MRSA cases (4%) were identified . One patient both had MRSA at admission and met our criteria for nosocomial MRSA . Of the remaining 39 patients, 11 had MRSA at admission and 28 had nosocomial MRSA . There were 17 wound infections, two cases of pneumonia, and two bloodstream infections . No deaths were attributed to MRSA sepsis . CONCLUSION: An MRSA control program including surveillance culturing, clinician feedback, flexible, site-specific isolation, and a list of known carriers is associated with a low rate of nosocomial MRSA in a pediatric burn unit. Eur J Epidemiol, 1994 Dec, 10(6), 743 - 8 Stability of genomic DNA fragment patterns in methicillin resistant Staphylococcus aureus (MRSA) during the course of intra- and interhospital spread; Witte W et al.; The analysis of genomic DNA fragment patterns has revealed as a powerful tool for strain discrimination in Staphylococcus aureus; for use as an epidemiological marker, stability during the course of an outbreak is an essential prerequisite . Genomic DNA fragment patterns (SmaI restriction, pulsed-field electrophoresis) of four different epidemic MRSA strains were compared along with intra- and interhospital and country-wide spread over more than 12 months in Germany . Strain I was isolated from infections in 8 hospitals . In one hospital a subclone arised which differed from the original strain by 4 fragments . Strain II was spread among 4 hospitals, isolates from three of these hospitals exhibited a variability of one to three fragments in the 150-200 kb range . Two hospitals in the Hannover-area were affected by strain III; in 17 isolates of this strain a variability up to three fragments was found in the 170-200 kb range . Strain IV was isolated from 19 cases of infections in 3 hospitals in Berlin . The fragment patterns were completely stable . When S . aureus strains are typed by genomic DNA fragment patterns, a variability in a definite range of molecular masses during the course of an epidemic should be taken into consideration. Rev Saude Publica, 1994 Dec, 28(6), 406 - 9 Staphylococcal food poisoning from cream-filled cake in a metropolitan area of south-eastern Brazil; Pereira ML et al.; Twelve people became ill with vomiting and diarrhoea approximately four hours after eating cake with a cream filling at a birthday party and on the day following . The cake had been prepared by a food handler who had long experience in preparing foods for such functions . Staphylococcus aureus that produced enterotoxin A was isolated from the nose, the fingernails, and a healed infection on the neck of the food handler, and from the cake . Enterotoxin A was detected in the remaining portion of the cake . The cake, while still warm, had been refrigerated for one hour after it was prepared before it was removed for the party; it was refrigerated after the party . The cake was large (6 kg) and hence it was not adequately cooled in the hour during which it was refrigerated before the party . The conclusion is that the cake was accidentally contaminated by the food handler and inadequately cooled before it was eaten. Mol Pharmacol, 1994 Dec, 46(6), 1048 - 55 Agonist-induced photoincorporation of a p-benzoylphenylalanine derivative of substance P into membrane-spanning region 2 of the Torpedo nicotinic acetylcholine receptor delta subunit; Blanton MP et al.; The neuropeptide substance P acts, at micromolar concentrations, as a noncompetitive antagonist of nicotinic acetylcholine receptors (AChRs) of both neuronal and muscle subtypes . The mechanism of this inhibition has been shown to be most consistent with stabilization of a nonconducting desensitized state of the AChR, via binding to a site distinct from both the agonist site and the high affinity noncompetitive antagonist site . We have used a radioiodinated photoreactive analogue of substance P, containing the amino acid p-benzoyl-L-phenylalanine in place of the Phe8 residue of substance P, to identify the sites of interaction of substance P within the Torpedo california AChR . AChR-rich membrane suspensions were photolabeled in the absence or presence of the agonist carbamylcholine and/or nonradioactive substance P, and incorporation into AChR subunits was assessed by autoradiography after sodium dodecyl sulfate-polyacrylamide gel electrophoresis . In the absence of agonist 125I incorporation was detected in each subunit and was insensitive to substance P, whereas in the presence of carbamylcholine there was a 2-fold increase in photoincorporation into the AChR delta subunit that was inhibited by the addition of an excess of substance P . The sites of specific photoincorporation in the delta subunit were initially mapped by use of Staphylococcus aureus V8 protease to a 14-kDa fragment extending from delta Ile-192 to Glu-280 . Further fragmentation of this 14-kDa fragment with trypsin and S . aureus V8 protease established that the sites of specific incorporation were restricted to the region delta Ser-253 to Glu-280, which contains the membrane-spanning region 2 that is known to form the lining of the ion channel . These results establish that in the presence of agonist at least a part of the undecapeptide substance P binds within the ion channel in the desensitized state of the AChR, and it is likely that the binding of substance P to this site is responsible for the action of substance P as a noncompetitive AChR antagonist. Leukemia, 1994 Dec, 8(12), 2083 - 94 Expression and regulation of CD30 ligand and CD30 in human leukemia-lymphoma cell lines; Gruss HJ et al.; The CD30 antigen was originally described as a specific surface marker for Hodgkin's lymphoma . Recent work established CD30 as a member of the tumor necrosis factor/nerve growth factor receptor superfamily whose ligand (CD30L) has also been cloned and expressed; CD30L is active as membrane-bound type II glycoprotein . Here, CD30L mRNA expression was studied in a panel of 102 continuous human leukemia-lymphoma cell lines and was found only in four Burkitt lymphoma, one Burkit-type acute lymphoblastic leukemia and one non-Hodgkin's lymphoma (NHL) cell line . The product of CD30L mRNA is expressed as a membrane protein on the surface of these malignant B-cell lines . Treatment of these cell lines with soluble CD27L, phorbol ester or staphylococcus aureus Cowan antigen resulted in the enhancement of cell surface CD30L protein expression . CD30L mRNA was not detected in normal unstimulated peripheral blood (PB) monocytes, monocyte-derived macrophages, or T-cells, but was detected in primary granulocytes; exposure to activating reagents induced and upregulated CD30L transcription in these different PB populations . While CD40 and CD30L surface protein expression on PB monocytes could be enhanced or induced by treatment with gamma-interferon, these cells remained negative for CD30, both at the mRNA and at the protein level . Similarly, PB monocyte-derived macrophages and granulocytes remained negative for CD30 mRNA and protein expression, regardless of stimulation . Only activated T-cells expressed CD30 mRNA and surface protein . CD30L-transfected cells and cells constitutively expressing CD30L delivered a similar stimulus for proliferation of the CD30+ Hodgkin's disease (HD)-derived cell line HDLM-2, but inhibited proliferation of the CD30+ large cell anaplastic lymphoma cell line KARPAS-299 . These data provide strong evidence for the involvement in growth regulation of recombinant and natural CD30L through its interaction with the CD30 receptor . Collectively, these data suggest that the CD30L-CD30 interaction has potent biological activity and might play a critical role in the immune response and pathogenesis of HD and some NHL, in particular Burkitt lymphomas. Biochemistry, 1994 Nov 22, 33(46), 13938 - 45 Identification of the neuronal acceptor in bovine cortex for ammodytoxin C, a presynaptically neurotoxic phospholipase A2; Krizaj I et al.; A specific, high-affinity binding site for ammodytoxin C in synaptic membranes from bovine cerebral cortex was detected and partially characterized . Equilibrium binding analysis revealed a single population of {125I}ammodytoxin C acceptors with the following binding parameters: Kd = 6.0 nM and Bmax = 5.7 pmol/mg membrane protein . Such binding was strongly inhibited by three ammodytoxins (A, B, and C) and by crotoxin B . Vipera berus berus phospholipase A2 was a weaker inhibitor; nontoxic phospholipase A2, ammodytin I2, and the myotoxic phospholipase A2 homologue, ammodytin L, both from Vipera ammodytes ammodytes venom, inhibited binding only at very high concentrations, whereas alpha-dendrotoxin, beta-bungarotoxin, and crotoxin A had no influence on the {125I}ammodytoxin C-specific binding . The ammodytoxin C neuronal binding site therefore overlaps, at least partially, with the neuronal acceptors for some of the related presynaptically neurotoxic phospholipases A2 (beta-neurotoxins) . {125I}-Ammodytoxin C was covalently attached to its acceptor by chemical cross-linking . Subsequent SDS-PAGE analysis followed by autoradiography revealed saturably labeled membrane components with apparent M(r) values of 51,000 (weaker band) and 53,000-56,000 (stronger band) . Pretreatment of synaptic membranes with Staphylococcus aureus V-8 proteinase and proteinase K, heat, or low pH decreased the {125I}ammodytoxin C-specific binding to various extents, but never abolished it completely . Membrane protein and certain phospholipids residing in its vicinity are therefore most likely involved in the binding of ammodytoxin C to bovine synaptic membranes. J Biol Chem, 1994 Nov 18, 269(46), 29121 - 8 Conformational differences in human apolipoprotein B-100 among subspecies of low density lipoproteins (LDL) . Association of altered proteolytic accessibility with decreased receptor binding of LDL subspecies from hypertriglyceridemic subjects; Chen GC et al.; We asked at what point in the metabolic cascade of very low density lipoproteins (VLDL) to low density lipoproteins (LDL) the accessibility of proteolytic cleavage sites in B-100 changes, and we evaluated the effect of hypertriglyceridemia on the proteolytic accessibility, secondary structure, and receptor-binding affinity of B-100 in LDL subspecies of varying density . Limited proteolysis with Staphylococcus aureus V8 protease and cathepsin D identified the density (about 1.033 g/ml) between two LDL subspecies, designated LDL-1 and -2, as the transition point during VLDL metabolism of both normolipidemic (N-) and hypertriglyceridemic (HTG-) subjects at which accessibility to protease attack changed in three peptide regions of B-100 . Hypertriglyceridemia greatly altered proteolytic accessibility of B-100 in the denser LDL subspecies . Specifically, B-100 in HTG-LDL exposed more cleavage sites than in N-LDL, including two novel sites, approximately 120 and approximately 130 kDa from the NH2 terminus in the small and dense subspecies (designated LDL-4, -4.5 or -5, d = 1.048-1.062 g/ml) . Analysis of circular dichroic spectra indicated no difference in helical content between B-100 in N- and HTG-LDL but showed a greater content of beta-structure in HTG-LDL . Binding affinity for the LDL receptor of human fibroblasts decreased markedly with increasing density among HTG-LDL subspecies (by approximately 50% for LDL-4.5 or -5) . We conclude that the changes in proteolytic accessibility observed between LDL-1 and -2 and in LDL-4, -4.5, or -5 indicate significant differences in local conformation of B-100 at specific peptide regions . The association of exposure of more cleavage sites, especially novel sites in the NH2-terminal regions, with greatly decreased receptor-binding affinity in LDL-4.5 or -5 suggests that altered local conformation in B-100 apart from the putative receptor-binding domain might affect interaction with the receptor. J Biol Chem, 1994 Nov 4, 269(44), 27246 - 50 Reduced methicillin resistance in a new Staphylococcus aureus transposon mutant that incorporates muramyl dipeptides into the cell wall peptidoglycan; Ornelas-Soares A et al.; Screening of a new Tn551 library constructed in the background of a highly methicillin-resistant Staphylococcus aureus strain identified a new insertion site located on the SmaI B-fragment of the chromosome that reduced the minimal inhibitory concentration of the parent (1600 micrograms/ml) to 25-50 micrograms/ml in the mutant and caused heterogeneous expression of resistance and abnormality in peptidoglycan composition (absence of the unsubstituted pentapeptide and incorporation of alanylglutamate- and alanylisoglutamate-containing muropeptides) . There was an accumulation of large amounts of the UDP-linked muramyl dipeptide in the cytoplasmic wall precursor pool of the mutant . Reduced (heterogeneous) antibiotic resistance and all the biochemical abnormalities were reproduced in genetic backcrosses by transduction with phage 80 alpha . Mutant RUSA235 appears to be impaired in the biosynthesis of the staphylococcal cell wall precursor muropeptide before the lysine addition step . We propose to provisionally call the gene inactivated in this mutant femF. Bull Tokyo Dent Coll, 1994 Nov, 35(4), 217 - 20 Detection of methicillin-resistant staphylococcus aureus in human saliva and on denture surfaces; Honma K et al.; The prevalence of Staphylococcus species and methicillin-resistant Staphylococcus aureus (MRSA) in human saliva and on denture surfaces was examined with selective media . A total of 166 saliva samples from dental students and individuals using dentures and 39 swab samples from denture surfaces were subjected in this study . The detected percentage of MRSA from saliva samples was 2.3% . Five out of 39 swab samples of denture surface contained MRSA or methicillin-resistant Staphylococcus epidermidis (MRSE) . The isolated MRSA and MRSE were resistant to beta-lactam antibiotics. J Appl Bacteriol, 1994 Nov, 77(5), 549 - 52 Staphylococcal growth and enterotoxins (A-D) and thermonuclease synthesis in the presence of dehydrated garlic; Gonzalez-Fandos E et al.; The inhibition of Staphylococcus aureus growth and enterotoxin and thermonuclease production by various concentrations of garlic (Allium sativum) was studied in BHI broth . The growth of Staph . aureus was inhibited by dehydrated garlic at levels of 1.5% (w/v) and over . Enterotoxins A, B and C1 were only detectable in broth containing < 1% of garlic while enterotoxin D was produced at a level of 2% . Garlic also inhibited thermonuclease (TNAse) production, complete inhibition being observed at levels > or = 1.5% . TNAse was not always detected when enterotoxin was present. J Antibiot (Tokyo), 1994 Nov, 47(11), 1266 - 72 Aldecalmycin, a new antimicrobial antibiotic from Streptomyces . I . Taxonomy, fermentation, isolation, physico-chemical and biological properties; Sawa R et al.; A new antibiotic, aldecalmycin, has been discovered in the culture broth of Streptomyces sp . MJ147-72F6 . Aldecalmycin was purified by solvent extraction, Diaion HP-20 chromatography, silica gel chromatography, Sephadex LH-20 chromatography, HPLC and centrifugal partition chromatography . The 1H and 13C NMR spectra of aldecalmycin showed the presence of keto-enol tautomers . Aldecalmycin is equipotent in inhibiting the growth of sensitive and methicillin-resistant Staphylococcus aureus (MRSA). J Antibiot (Tokyo), 1994 Nov, 47(11), 1250 - 7 Pyrroindomycins, novel antibiotics produced by Streptomyces rugosporus sp . LL-42D005 . I . Isolation and structure determination; Ding W et al.; Pyrroindomycins A and B were isolated from fermentations of culture LL-42D005, a strain of Streptomyces rugosporus . Pyrroindomycins possess potent antimicrobial activities against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci . Their structures have been determined by using 1- and 2-D NMR, mass spectroscopy and chemical degradations . Pyrroindomycins are the first natural products that contain the highly unsaturated pyrroloindole moiety. Br J Dermatol, 1994 Nov, 131(5), 717 - 9 Juvenile pityriasis rubra pilaris associated with hypogammaglobulinaemia and furunculosis; Castanet J et al.; We report a case of follicular keratosis with inflammatory changes, consistent with a diagnosis of atypical juvenile pityriasis rubra pilaris . An unusual feature was the occurrence of severe Staphylococcus aureus folliculitis and furunculosis, a phenomenon rarely encountered in pityriasis rubra pilaris and the other follicular keratoses . Standard antibiotic and antiseptic treatment for chronic S . aureus infection was ineffective . The patient was subsequently found to have hypogammaglobulinaemia, and treatment with human polyvalent immunoglobulin infusions was successful in eradicating the sepsis . It is therefore probable that the hypogammaglobulinaemia played a pathogenic role in the development of cutaneous sepsis. Medicine (Baltimore), 1994 Nov, 73(6), 299 - 305 Pacemaker endocarditis . Report of 44 cases and review of the literature; Arber N et al.; We conducted a retrospective study to characterize the clinical course, microbiologic spectrum, and risk factors for endocarditis and for associated mortality in a large series of patients with documented pacemaker endocarditis . Using a computerized search through the medical records of 10 major hospitals in Israel from 1982 to 1992, and carefully reviewing the charts, we identified 44 patients with pacemaker endocarditis . The cases were categorized as definite (n = 25), probable (n = 12), or possible (n = 7) infective endocarditis based on strict case definition . Fever and chills were the most common symptoms . Increased ESR, leukocytosis, microscopic hematuria, and anemia were the most common laboratory findings . A relatively high proportion of the patients were diabetic . The most common source of endocarditis was infection acquired by the placement procedure or infection of the pacemaker pouch . Demographic, clinical, and laboratory features were similar to those of endocarditis patients of a similar age range without pacemakers, although the frequency of fever and chills was higher in our patients than in those patients and splenomegaly, vascular embolic phenomena, and new or changing murmurs were rare in our patients . The major pathogens were Staphylococcus aureus and Staphylococcus epidermidis, similar to other series of pacemaker-associated bacteremia and similar to the microbiologic findings of early prosthetic-valve endocarditis . However, this microbiologic profile is different from that of native-valve endocarditis . Although the present series did not show a statistically significant advantage to electrode removal over conservative treatment, when analyzed together with pooled data from other studies, it suggests that the surgical approach is preferable.(ABSTRACT TRUNCATED AT 250 WORDS) J Bone Joint Surg Br, 1994 Nov, 76(6), 955 - 9 Influence of type of medullary nail on the development of local infection . An experimental study of solid and slotted nails in rabbits; Melcher GA et al.; Any operation involving the implantation of a foreign body increases the risk of infection . The implant material and its surface, the dead space, and any necrosis or vascular changes play a significant role in susceptibility to infection . We investigated the effect of the dead space in an intramedullary nail on the rate of local infection . We inoculated the intramedullary cavities of rabbit tibiae with various concentrations of a human pathogen, of Staphylococcus aureus strain, and then inserted either a solid or a hollow slotted stainless-steel nail . We found a significantly higher rate of infection after use of the slotted nail (59%) than after the solid nail (27%) (p < 0.05). Am J Physiol, 1994 Nov, 267(5 Pt 1), L578 - 84 Binding of surfactant protein A to C1q receptors mediates phagocytosis of Staphylococcus aureus by monocytes; Geertsma MF et al.; During both steady-state conditions and inflammatory reactions in the lower airways, monocytes migrate to the alveoli where they come into contact with surfactant . Surfactant is composed of phospholipids, neutral lipids, and specific proteins, and its main function is to reduce surface tension in the alveoli . The most abundant glycoprotein surfactant protein A (SP-A) affects the structure, function, and metabolism of pulmonary surfactant . The aim of the present study was to determine whether SP-A plays a role in the antibacterial activities of human monocytes and whether this is mediated by a receptor for SP-A on these cells . The results showed that SP-A binds to both Staphylococcus aureus and monocytes and mediates the phagocytosis of the bacteria by these cells . SP-A does not stimulate the intracellular killing of bacteria by monocytes, and SP-A-opsonized S . aureus do not induce the production of reactive oxygen intermediates . SP-A binds to the C1q receptor (C1qR) on monocytes, since its binding was inhibited by C1q and the SP-A-enhanced association of S . aureus with these cells was completely abolished when monocytes were adherent to surfaces coated with C1q or anti-C1qR monoclonal antibody . Furthermore, the binding of SP-A to monocytes results in an increased intracellular concentration of adenosine 3',5'-cyclic monophosphate . Together, these results demonstrate that C1qR mediates the phagocytosis of SP-A-opsonized S . aureus by monocytes. Surg Neurol, 1994 Nov, 42(5), 424 - 7 Spinal subdural abscess: successful treatment with limited drainage and antibiotics in a patient with AIDS; Sathi S et al.; The authors present a case of Staphylococcus aureus spinal subdural abscess in a patient with AIDS . Although complete surgical drainage has been strongly advocated in the literature, this patient made a complete neurologic and symptomatic recovery and radiographically demonstrated resolution of the abscess with only limited surgical drainage and parenteral antibiotics . Magnetic resonance imaging findings of this unusual lesion are discussed . Relevant literature in the management of spinal subdural abscesses is reviewed. Surg Neurol, 1994 Nov, 42(5), 417 - 23 HIV status does not affect microbiologic spectrum or neurologic outcome in spinal infections; Heary RF et al.; The impact of human immunodeficiency virus (HIV) on the clinical presentations, causative organisms, and neurologic outcomes of patients with spinal infections is reviewed . Thirty-two patients with spinal epidural abscesses, vertebral osteomyelitis, or both were treated at an urban hospital over a 42-month period . Thirteen of these patients were confirmed by serologic analysis to be HIV seropositive . The diagnoses were confirmed by 30 open surgical procedures (14 anterior, 16 posterior) and seven percutaneous biopsies . Twenty-seven intraoperative cultures were positive and the remaining three patients had positive blood cultures prior to the surgical procedure . In both the HIV (+) and HIV (-) groups, Staphylococcus aureus predominated as the causative organism (overall rate: 72%) . Mycobacterium tuberculosis was the second most common organism . The clinical presentations in both groups were similar with pain as the most frequent symptom and objective neurologic abnormalities on physical examination in 29 of the 32 patients (91%) . The results of this analysis show that the clinical presentations and organisms cultured do not differ depending upon a concurrent HIV infection . Furthermore, the ultimate neurologic outcome of patients with spinal infections depends on their neurologic status at the time of treatment and not on their HIV status. J Gen Virol, 1994 Nov, 75 ( Pt 11), 2897 - 909 Structural protein relationships among eastern equine encephalitis viruses; Strizki JM et al.; We have re-evaluated the relationships among the polypeptides of eastern equine encephalitis (EEE) viruses using SDS-PAGE and peptide mapping of individual virion proteins . Four to five distinct polypeptide bands were detected upon SDS-PAGE analysis of viruses: the E1, E2 and C proteins normally associated with alphavirus virions, as well as an additional more rapidly-migrating E2-associated protein and a high M(r) (HMW) protein . In contrast with previous findings by others, the electrophoretic profiles of the virion proteins of EEE viruses displayed a marked correlation with serotype . The protein profiles of the 33 North American (NA)-serotype viruses examined were remarkably homogeneous, with variation detected only in the E1 protein of two isolates . In contrast, considerable heterogeneity was observed in the migration profiles of both the E1 and E2 glycoproteins of the 13 South American (SA)-type viruses examined . Peptide mapping of individual virion proteins using limited proteolysis with Staphylococcus aureus V8 protease confirmed that, in addition to the homogeneity evident among NA-type viruses and relative heterogeneity among SA-type viruses, the E1 and E2 proteins of NA- and SA-serotype viruses exhibited serotype-specific structural variation . The C protein was highly conserved among isolates of both virus serotypes . Endoglycosidase analyses of intact virions did not reveal substantial glycosylation differences between the glycoproteins of NA- and SA-serotype viruses . Both the HMW protein and the E2 protein (doublet) of EEE virus appeared to contain, at least in part, high-mannose type N-linked oligosaccharides . No evidence of O-linked glycans was found on either the E1 or the E2 glycoprotein . Despite the observed structural differences between proteins of NA- and SA-type viruses, Western blot analyses utilizing polyclonal antibodies indicated that immunoreactive epitopes appeared to be conserved. J Exp Med, 1994 Nov 1, 180(5), 1675 - 83 Characterization and biological properties of a new staphylococcal exotoxin; Ren K et al.; Staphylococcus aureus strain D4508 is a toxic shock syndrome toxin 1-negative clinical isolate from a nonmenstrual case of toxic shock syndrome (TSS) . In the present study, we have purified and characterized a new exotoxin from the extracellular products of this strain . This toxin was found to have a molecular mass of 25.14 kD by mass spectrometry and an isoelectric point of 5.65 by isoelectric focusing . We have also cloned and sequenced its corresponding genomic determinant . The DNA sequence encoding the mature protein was found to be 654 base pairs and is predicted to encode a polypeptide of 218 amino acids . The deduced protein contains an NH2-terminal sequence identical to that of the native protein . The calculated molecular weight (25.21 kD) of the recombinant mature protein is also consistent with that of the native molecules . When injected intravenously into rabbits, both the native and recombinant toxins induce an acute TSS-like illness characterized by high fever, hypotension, diarrhea, shock, and in some cases death, with classical histological findings of TSS . Furthermore, the activity of the toxin is specifically enhanced by low quantities of endotoxins . The toxicity can be blocked by rabbit immunoglobulin G antibody specific for the toxin . Western blotting and DNA sequencing data confirm that the protein is a unique staphylococcal exotoxin, yet shares significant sequence homology with known staphylococcal enterotoxins, especially the SEA, SED, and SEE toxins . We conclude therefore that this 25-kD protein belongs to the staphylococcal enterotoxin gene family that is capable of inducing a TSS-like illness in rabbits. J Leukoc Biol, 1994 Nov, 56(5), 661 - 5 Isolation of antimicrobial peptides from avian heterophils; Evans EW et al.; Five bactericidal peptides (chicken heterophil peptides CHP1 and CHP2; turkey heterophil peptides THP1, THP2, and THP3) were purified from avian heterophil granules . All peptides were cationic and rich in cysteine, arginine, and lysine . The complete amino acid sequence, consisting of 39 amino acids, was determined for CHP1 . This peptide had a molecular weight of 4481 as determined by mass spectrometry . Partial NH2-terminal amino acid sequences were obtained for the remaining peptides . Both chicken peptides and THP1 shared sequence homology at 22 residues and a cysteine motif which was similar to that of bovine beta-defensins . THP2 and THP3 were homologous to each other but were not homologous to the other three and had a unique cysteine motif . Peptides CHP1, CHP2, and THP1 killed Staphylococcus aureus and Escherichia coli in vitro, whereas THP2 and THP3 killed only S . aureus in vitro. J Clin Invest, 1994 Nov, 94(5), 1815 - 22 Diminished virulence of a sar-/agr- mutant of Staphylococcus aureus in the rabbit model of endocarditis; Cheung AL et al.; Microbial pathogenicity in Staphylococcus aureus is a complex process involving a number of virulence genes that are regulated by global regulatory systems including sar and agr . To evaluate the roles of these two loci in virulence, we constructed sar-/agr- mutants of strains RN6390 and RN450 and compared their phenotypic profiles to the corresponding single sar- and agr- mutants and parents . The secretion of all hemolysins was absent in the sar-/agr- mutants while residual beta-hemolysin activity remained in single agr- mutants . The fibronectin binding capacity was significantly diminished in both single sar- mutants and double mutants when compared with parents while the reduction in fibrinogen binding capacity in the double mutants was modest . In the rabbit endocarditis model, there was a significant decrease in both infectivity rates and intravegetation bacterial densities with the double mutant as compared to the parent (RN6390) at 10(3)-10(6) CFU inocula despite comparable levels of early bacteremia among various challenge groups . Notably, fewer bacteria in the double mutant group adhered to valvular vegetations at 30 min after challenge (10(6) CFU) than the parent group . These studies suggest that both the sar and agr loci are involved in initial valvular adherence, intravegetation persistence and multiplication of S . aureus in endocarditis. Exp Parasitol, 1994 Nov, 79(3), 468 - 79 Leishmania major: association of the differentially expressed gene B protein and the surface lipophosphoglycan as revealed by membrane capping; Pimenta PF et al.; The lipophosphoglycan (LPG) of Leishmania promastigotes forms a dense glycocalyx which effectively covers the entire surface of the cell, and which undergoes structural modifications during the differentiation of promastigotes to the infective or metacyclic stage . Recently, the first protein marker for metacyclic promastigotes of Leishmania major has been characterized . This protein, termed gene B protein, is located on the cell surface, yet it lacks any hydrophobic sequence for membrane attachment . It does contain an unusual amino acid repeat that is related to the peptidoglycan binding domain of protein A from Staphylococcus aureus, suggesting that the protein might interact with metacyclic LPG via this domain for attachment to the cell . We have studied the distribution of LPG, gene B protein, and the major surface protease, gp63, by labeling them with immunogold or immunofluorescence prior to and during capping events . Thin sections of double-labeled parasites revealed that the gene B protein-gold particles were colocalized with the LPG-gold particles in the LPG capping structures at the extremities of the cell . Cocapping of LPG and gene B protein was also observed with two-color fluorescence . No similar redistribution was seen in gp63 or with integral membrane proteins . In contrast to the gene B protein, gp63 could only be immunogold labeled on the metacyclic surface after capping and shedding of the LPG, providing further that it and other membrane-associated proteins are normally buried under the LPG coat . The unusual surface exposure of the gene B protein is consistent with its hydrophilic and LPG binding properties, which allow it to become incorporated into the cell coat and to localize to the most external aspects of the cell. EMBO J, 1994 Nov 1, 13(21), 5245 - 51 Replication-specific conversion of the Staphylococcus aureus pT181 initiator protein from an active homodimer to an inactive heterodimer; Rasooly A et al.; The Staphylococcus aureus rolling circle plasmid pT181 regulates its replication by controlling the synthesis of its initiator protein RepC . RepC is inactivated during pT181 replication by the addition of an oligodeoxynucleotide, giving rise to a new form, RepC* . We analyzed RepC and RepC* in four classes of mutants: plasmid copy number mutants, two classes of RepC mutants affecting different portions of the protein and oriC (origin) mutants . We have found that in the cell with wild-type RepC there are similar relative amounts of RepC and RepC*, regardless of copy number, and that the conversion of RepC to RepC* is replication dependent . Genetic and biochemical evidence is presented that RepC functions as a dimer and that during replication the RepC homodimer is converted to the RepC/RepC* heterodimer. Biochemistry, 1994 Nov 1, 33(43), 12844 - 51 Mutations affecting the activity of toxic shock syndrome toxin-1; Deresiewicz RL et al.; Toxic shock syndrome toxin-1 (TSST-1), the potent staphylococcal exoprotein linked to most cases of the toxic shock syndrome, is a V beta-restricted T-cell mitogen (a so-called "superantigen") . TSST-ovine (TSST-O) is a natural variant of TSST-1, and is produced by certain ovine mastitis-associated strains of Staphylococcus aureus . Compared to TSST-1, TSST-O is only weakly mitogenic for leporine or murine splenocytes . It differs from TSST-1 at 7 amino acid residues over its 194 amino acid length . Terminus shuffling between the two proteins has suggested that their C-terminal differences (T69, Y80, E132, and I140 in TSST-1; 169, W80, K132, and T140 in TSST-O) are in part responsible for their discrepant mitogenic properties . In order to explore further the functional consequences of altering TSST-1 at residues 132 and 140, we engineered point mutants of TSST-1 at those positions . The mutant proteins were purified to homogeneity from culture supernants of a nontoxigenic strain of S . aureus using a combination of ultrafiltration, liquid-phase isoelectric focusing, and ion-exchange chromatography . The mutants retained global structural integrity as evidenced by circular dichroism spectroscopy, their preserved resistance to trypsin digestion, and their preserved binding to a neutralizing murine monoclonal antibody . The mutants were then tested for mitogenicity for human T-cells: The mutant I140T was approximately as active as wild-type TSST-1, while the mutant E132D was about 10-fold attenuated . On the other hand, the mutants E132A or E132K were each at least 1000-fold attenuated.(ABSTRACT TRUNCATED AT 250 WORDS) J Am Acad Dermatol, 1994 Nov, 31(5 Pt 1), 746 - 54 Cutaneous findings in HIV-1-positive patients: a 42-month prospective study . Military Medical Consortium for the Advancement of Retroviral Research (MMCARR); Smith KJ et al.; BACKGROUND: Cutaneous disease is common in patients infected with HIV-1 . OBJECTIVE: The aim of our study was to identify cutaneous markers associated with HIV-1 infection and disease progression as measured by Walter Reed (WR) stage . METHODS: For 42 months we have observed 912 HIV-1-positive patients in all WR stages . All patients had an extensive past and present medical history taken as well as a complete physical examination, periodic visits, and appropriate diagnostic procedures . RESULTS: Increasing dryness of the skin and seborrheic dermatitis are early findings in a large percentage of patients in WR stage 1; the occurrence and severity of both conditions increase with disease progression . Tinea infections, condylomata acuminata, and verrucae are seen early, but with disease progression, although there is no clear increase in occurrence, these infections become more diffuse and resistant to treatment . Flares in acne vulgaris and folliculitis show a peak occurrence in early and mid-stage disease with a decreased occurrence in late-stage disease . Herpes simplex infections, oral candidiasis, molluscum contagiosum, Staphylococcus aureus infections, and oral hairy leukoplakia show a marked increase in occurrence with advanced disease . Conditions that have a statistically significant association with disease progression as measured by a change in a stage include drug eruptions, seborrheic dermatitis, oral candidiasis, oral hairy leukoplakia, molluscum contagiosum, herpes zoster, and hyperpigmentation (nail, oral, skin) . CONCLUSION: The most frequent and persistent cutaneous disorders were asteatosis (with or without asteatotic eczema) and seborrheic dermatitis . Conditions that were associated with a change in WR stage include drug eruptions, seborrheic dermatitis, oral candidiasis, oral hairy leukoplakia, molluscum contagiosum, herpes zoster, and hyperpigmentation . In addition to Kaposi's sarcoma, patients with HIV-1 disease have an increased potential for the development of both cutaneous epithelial and probably melanocytic malignancies . Epithelial tumors were seen in patients in all stages of disease. Plasmid, 1994 Nov, 32(3), 295 - 305 A conjugative transfer system for the rumen bacterium, Butyrivibrio fibrisolvens, based on Tn916-mediated transfer of the Staphylococcus aureus plasmid pUB110; Clark RG et al.; A limitation of genetic studies of the rumen bacterium, Butyrivibrio fibrisolvens, has been the availability of suitable vectors and transfer systems . Using the conjugative tetracycline resistant transposon, Tn916, the Staphylococcus aureus plasmid, pUB110, and the pUB110-based shuttle vector, pUBLRS, a conjugative transfer system was developed for B . fibrisolvens . B fibrisolvens donor strains H17c2 and H17c12, containing Tn916 and pUB110 or pUBLRS, respectively, were used in mating experiments with selected B . fibrisolvens strains . Kanamycin resistant transconjugants, containing pUB110, of strains 193, 194, and 195 were detected at a combined average frequency of 7.78 x 10(-7) per donor and 1.11 x 10(-5) per recipient . Transconjugants of strains 193 and 194, containing pUBLRS, were detected at an average frequency of 1.22 x 10(-6) per donor and 4.70 x 10(-8) per recipient . Southern hybridization analysis confirmed the presence of pUB110 and pUBLRS in transconjugants . Results indicated that Tn916 was necessary for mobilization of pUB110 as transconjugants were not detected when the transposon was absent from the donor strains . The ability to mobilize pUB110 and pUBLRS between B . fibrisolvens strains provides a conjugative transfer system that circumvents problems encountered with electroporation. J Pharm Pharmacol, 1994 Nov, 46(11), 892 - 5 Flavanones with potent antibacterial activity against methicillin-resistant Staphylococcus aureus; Iinuma M et al.; With the therapeutic concept of using the defensive ability of plants against microbial infections, phytoalexin, an antimicrobial phytochemical was studied for its ability to inhibit the growth of methicillin-resistant Staphylococcus aureus (MRSA) . Extracts from Sophora exigua (Leguminosae) were fractionated by serial chromatography and the anti-MRSA activity of each fraction was determined by the agar-plate method . Among the active isolates, 5,7,2',6'-tetrahydroxy-6-isoprenyl-8-lavandulyl-4'-methox yflavanone (exiguaflavanone D) completely inhibited the growth of all the MRSA strains examined at the concentration of 1.56-6.25 micrograms mL-1, and 5, 2',6'-trihydroxy-8-lavandulyl-7-methoxy-flavanone (exiguaflavanone B) inhibited at a concentration of 50 micrograms mL-1 . This former compound is expected to be a phytotherapeutic agent for MRSA infections as an alternative to conventional antibiotics with unwanted side-effects or the appearance of antibiotic-resistant bacteria. Clin Infect Dis, 1994 Nov, 19(5), 970 - 2 Infectious complications associated with temporary epidural catheters; Pegues DA et al.; To identify cases of deep-tissue or local infection associated with temporary epidural catheters, we reviewed medical records from 1980 through 1992 and prospectively followed up patients with temporary epidural catheters from January 1993 through June 1993 who were hospitalized at a large, tertiary referral hospital . We identified seven cases of temporary epidural catheter-associated infection, including three cases of deep-tissue infection (paraspinal muscle abscess, epidural abscess, and meningitis) and four cases of local infection . The infections were diagnosed 2 days to 22 days following insertion of the epidural catheter . Staphylococcus aureus was isolated in four (57%) of the cases . All seven patients were treated with removal of the catheter and antimicrobial therapy; three patients also required surgical drainage for management of deep-tissue infection . Our findings emphasize the importance of daily inspection and prompt removal of temporary epidural catheters if infection is suspected. Diagn Microbiol Infect Dis, 1994 Nov, 20(3), 171 - 4 Comparison of mupirocin susceptibility of nasal and nonnasal Staphylococcus aureus isolates; Utrup LJ et al.; Susceptibilities of 414 nasal and 586 nonnasal Staphylococcus aureus isolates, both methicillin resistant and methicillin susceptible, to the topical antimicrobial agent mupirocin were compared . A susceptibility of 99.1% was observed for the 1000 isolates . Nasal and nonnasal isolates showed a similar 90% minimum inhibitory concentration (MIC90) and statistically equivalent percent susceptibilities. FEMS Immunol Med Microbiol, 1994 Nov, 10(1), 47 - 53 Vaccination with Staphylococcus aureus fibrinogen binding proteins (FgBPs) reduces colonisation of S . aureus in a mouse mastitis model; Mamo W et al.; A mouse mastitis model was used to study the effect of vaccination with fibrinogen binding proteins and collagen binding protein from Staphylococcus aureus against challenge infection with S . aureus . The mice vaccinated with fibrinogen binding proteins showed reduced rates of mastitis compared with controls . Gross examination of challenged mammary glands of mice showed that the glands of mice immunized with fibrinogen binding proteins developed mild intramammary infection or had no pathological changes compared with glands from control mice . Histopathological examination of tissue sections from challenged glands showed that most glands from mice vaccinated with fibrinogen binding protein developed disseminated necrosis or had no pathological changes . A significantly reduced number of bacteria could be recovered in the glands from mice immunized with fibrinogen binding proteins as compared with controls . In a similar study, immunization of mice with collagen binding protein did not induce protection against challenge infection with S . aureus. Antimicrob Agents Chemother, 1994 Nov, 38(11), 2590 - 8 Reassessment of the number of auxiliary genes essential for expression of high-level methicillin resistance in Staphylococcus aureus; de Lencastre H et al.; A new transposon library constructed in the background of the highly and homogeneously methicillin-resistant Staphylococcus aureus strain COL yielded 70 independent insertional mutants with reduced levels of antibiotic resistance . Restriction analysis with HindIII, EcoRV, EcoRI, and PstI and then Southern hybridization with probes for the transposon and for the femA-femB gene demonstrated that 41 of the 70 Tn551 mutants carried distinct and novel, as yet undescribed insertion sites, all of which were outside of the mecA gene and were also outside the already-characterized auxiliary genes femA, femB, femC, and femD . All previously described Tn551 mutations of this type were in genes located either on SmaI fragment A or SmaI fragment I . In contrast, inserts of the new library were located in 7 of the 16 SmaI chromosomal fragments, fragments A, B, C, D, E, F, and I . In all of the mutants, expression of methicillin resistance became heterogeneous, and the MIC for the majority of cells was reduced (1.5 to 200 micrograms ml-1) from the homogeneous methicillin MIC (1,600 micrograms ml-1) of the parental cells . Although identification of the exact number of genes inactivated through the new set of transposon inserts will require cloning and sequencing, a rough estimate of this number from mapping data suggests a minimum of at least 10 to 12 new genetic determinants, all of which are needed together with femA, femB, femC, and femD for the optimal expression of methicillin resistance. Antimicrob Agents Chemother, 1994 Nov, 38(11), 2568 - 71 Altered production of penicillin-binding protein 2a can affect phenotypic expression of methicillin resistance in Staphylococcus aureus; Hackbarth CJ et al.; Altered production of penicillin-binding protein 2a (PBP 2a) may affect the phenotypic expression of resistance in methicillin-resistant Staphylococcus aureus (MRSA) . COL, an MRSA strain that constitutively produces PBP 2a, was transformed with a recombinant plasmid containing the two beta-lactamase regulatory genes, blaI and blaR1, with either the beta-lactamase gene, blaZ, or a truncated blaZ . Both of the transformed MRSA strains now produced an inducible PBP 2a, and the MICs of nafcillin, methicillin, and imipenem for these strains were similar to those for the parental strain . A mutation in blaR1 that resulted in the complete repression of PBP 2a production altered the phenotypic expression of methicillin resistance in that strain, as evidenced by efficiency-of-plating experiments . Rather than being homogeneously resistant like COL, the blaR1 mutant strain now appeared to have a small resistant subpopulation . Gene products that regulate PBP 2a production may contribute to the organism's expression of methicillin resistance, but additional chromosomally located factors are required. Anticancer Res, 1994 Nov-Dec, 14(6B), 2633 - 6 Effects of amino and imino acridines on tumor necrosis factor production by human leukocytes; Mandi Y et al.; Tumor necrosis factor (TNF) is a multifunctional cytokine with diverse effects on different cells and tissues . The biological activity of TNF is described on the basis of its cytotoxic action in vivo and in vitro . Different acridines were systematically synthesized and their effects were tested on endotoxin and Staphylococcus aureus-induced TNF production by human leukocytes . 9-aminobutylacridine and 9-ethylaminoacridine totally abrogated the TNF production of leucocytes at a concentration of 3.5 microM, whereas 9-imino -10-butylacridine and 9-imino-10-ethylacridine exerted only a 50% inhibition in the same concentration . Derivatives designated as 9-amino-(2-dimethylamino-ethyl)-acridine and 9-imino-10-(2-dimethylamino-ethyl)-acridine in a concentration of 7 microM exerted only a 30% and a 10% inhibition respectively . A significant modulation of TNF production was not observed when other alkylated derivatives in this series were applied . The TNF-mediated cytotoxic effect of monocytes against WEHI cells was also reduced by the most effective compounds . The acridines did not interfere with the expression of CD 14 molecules on monocytes . The exact mechanism of the suppression of TNF synthesis by acridines remains to be elucidated, but might be useful in the screening and evaluation of their anticancer properties and antimalarial effects. Mol Cell Endocrinol, 1994 Nov, 105(2), 217 - 26 Arginine vasopressin (AVP) causes the reversible phosphorylation of the myristoylated alanine-rich C kinase substrate (MARCKS) protein in the ovine anterior pituitary: evidence that MARCKS phosphorylation is associated with adrenocorticotropin (ACTH) secretion; Liu JP et al.; We have recently shown that AVP causes a protein kinase C (PKC)-dependent increase in ACTH release and biosynthesis in ovine anterior pituitary cells . In these cells, AVP also causes the translocation of PKC from the cytosol to the cell membrane which is maximal at 5 min, but the intracellular events distal to protein kinase C activation that underlie ACTH secretion have not been well characterized to date . Since the MARCKS protein has been implicated in neurosecretion and is phosphorylated by PKC in synaptosomes, studies were carried out to determine whether AVP might cause MARCKS phosphorylation in the ovine anterior pituitary, and to determine whether this phenomenon might be temporally correlated with PKC translocation and the release of ACTH . When cytosolic fractions of rat brain, ovine anterior pituitary, and cultured ovine anterior pituitary cells were incubated with purified PKC, several proteins were phosphorylated including those in the region of 83-85 kDa . After precipitation of the proteins with 40% acetic acid, the 83-85 kDa phosphoproteins were selectively recovered in the acid soluble phase . Phosphopeptide maps of either the 83 or 85 kDa proteins were generated with Staphylococcus aureus V8 protease and revealed 13 and 9 kDa phosphopeptides, which are characteristic of the authentic MARCKS protein . An identical phosphopeptide map was also obtained when the MARCKS protein was selectively extracted from intact 32P-labeled anterior pituitary cells . MARCKS phosphorylation was markedly increased when ovine anterior pituitary cells were exposed to 1 microM phorbol 12-myristate 13-acetate (PMA) . When the cells were exposed to 1 microM AVP, MARCKS phosphorylation increased at 15 s and reached the maximal plateau value at 30 s . MARCKS phosphorylation then started to diminish at 2 min, and baseline levels were attained by 10 min . In the same cells, AVP stimulated ACTH release in a biphasic manner-during the first 30 s, there resulted a rapid burst of ACTH secretion that was followed by a slower, but sustained rate of secretion . We conclude that: (1) AVP causes a rapid, and reversible, phosphorylation of the MARCKS protein in the ovine anterior pituitary; (2) since the AVP-induced increase in MARCKS phosphorylation occurs much earlier in these cells than does PKC trans-location, MARCKS phosphorylation may provide a more sensitive index of the onset of PKC activation than the translocation assay; (3) the close temporal association between MARCKS phosphorylation and the rapid early release of ACTH suggests that MARCKS phosphorylation may be involved in the initial intracellular events that underly exocytosis of the hormone. East Afr Med J, 1994 Nov, 71(11), 736 - 8 Absence of neutropenia in African patients with AIDS and associated pyomyositis; Ansaloni L et al.; The association between AIDS and pyomyositis was recently pointed out in temperate and tropical countries . In Western countries, the patients affected by pyomyositis associated with AIDS in most cases are neutropenic . We compare a group of 17 patients with pyomyositis and AIDS living in temperate climates from the literature, and 11 patients affected by the same association seen by us in northern Uganda . The patients from Western countries were significantly more neutropenic and their mean of the neutrophil count was significantly lower when compared with our group . We suggest that the defective neutrophil function associated with HIV infection play a major role in the pathogenesis of pyomyositis in our patientsPIP: Pyomyositis is an infection of the skeletal muscle mostly caused by Staphylococcus aureus . Patients in Western countries affected by pyomyositis associated with AIDS tend to be neutropenic . The objective of this study was to find out whether neutropenia is a common feature of African AIDS patients with pyomyositis; therefore, an African sample was compared with a sample of Western patients obtained from the international literature . During January-December 1993, 30 patients with pyomyositis were admitted to the Surgical Ward of Dr . Ambrosoli Memorial Hospital in Kalongo, Uganda . 11 (36.6%) were found positive for HIV and were in stage IV of the disease . The mean age was 28 years, 3 were females and 8 were males . The neutrophil count was performed before the surgical evacuation of muscle abscess . In the international literature, eight reports of 17 patients with AIDS and pyomyositis were located during the period of 1988-92 . All were males in stage IV with a mean age of 37.3 years; 8 were treated with zidovudine . When the neutrophil count was below 3000/cu . mm, the patient was considered neutropenic . 12 out of 17 Western patients were neutropenic, as opposed to only 1 out of 11 Ugandan patients (p 0.01) . The mean neutrophil count of Western patients was 3547; that of Ugandan patients was 9077 (p 0.01) . Neutrophils are primary effector cells in host defense against staphylococcal infections such as pyomyositis; hence, it has been suggested that neutropenia contributes to the development of pyomyositis in AIDS patients . Neutrophils from AIDS patients were demonstrated to be defective in their ability to kill Staphylococcus aureus in vitro, compared with neutrophils from seronegative controls . Therefore, the defective neutrophil function associated with HIV infection contributed to the development of pyomyositis in Ugandan patients, among whom neutropenia was not common . Infect Control Hosp Epidemiol, 1994 Nov, 15(11), 703 - 9 Colonization and transmission of high-level gentamicin-resistant enterococci in a long-term care facility; Chenoweth CE et al.; OBJECTIVES: To assess the prevalence of high-level gentamicin-resistant enterococcus (HGRE) colonization, transmission patterns, and spectrum of illness among residents of a long-term care facility . DESIGN: Monthly surveillance for HGRE colonization of wounds, rectum, and perineum over a 1-year period . SETTING: A Veterans Affairs long-term care facility attached to an acute-care facility . PATIENTS: All 341 patients in the facility during the observation period . RESULTS: Over the 1-year period, 120 patients (35.2%) were colonized with HGRE at least once, with an overall monthly colonization rate of 20 +/- 1.5% . HGRE were isolated from rectum (12.8%), wounds (11.7%), and perineum (9.3%) . Patients with the poorest functional status had the highest rate of colonization (P < 0.0005) . HGRE-colonized patients were more likely to be colonized with methicillin-resistant Staphylococcus aureus (51% versus 25%; P < 0.0005) . Seventy-four patients (21.7%) were colonized at admission or at the start of the study . Another 46 patients (13.5%) acquired HGRE during the study, including 36 who acquired HGRE while in the long-term care facility and 10 who were positive when transferred back from the acute-care hospital . Based on plasmid profiles, only two patients appeared to have isolates similar to those of current or previous roommates . Carriage of HGRE was transient in most cases . Only 20 patients were colonized for 4 or more months, and those patients usually carried different strains intermittently . Infections were infrequent, occurring in only 4.1% of total patients . CONCLUSIONS: In our long-term care facility, HGRE were endemic, and new acquisition of HGRE occurred frequently . However, only two patients had evidence of acquisition from a roommate, suggesting that cross-infection from a roommate was not a major route of spread of HGRE. Rinsho Byori, 1994 Nov, 42(11), 1182 - 7 {Detection of methicillin-resistant Staphylococcus aureus using PCR and non-radioactive DNA probes: IV . Mutational sequences in the region upstream of the mec A gene in clinical staphylococcal strains}; Yamashita K et al.; Expression of mec A is regulated by two regulatory genes (mecR1 and mecI); the intact regulatory genes exist in the N315 strain of methicillin-sensitive S . aureus, whereas the repressor gene (mecI) is deleted in the MR108 strain of methicillin-resistant S . aureus (MRSA) . However, our previous findings that clinical strains of N315 type were methicillin-resistant suggest possible mutations within the operator region, by which the inhibitory effect of MecI is released to induce the constitutive expression of mec A . Therefore, we analyzed nucleotide sequences in the region upstream of the mec A gene and in the regulatory genes, by using PCR-direct sequencing and ASO probes . Analysis of selected clinical strains by ASO probes revealed that 74% of MRSA and 36% of MRSE (methicillin-resistant S . epidermidis) lacked the mecI gene whereas 26% of MRSA and 64% of MRSE possessed the intact regulatory genes . Furthermore, PCR-direct sequencing identified four different mutations in the operator nucleotide sequence and one mutation in Shine-Dalgarno sequence . Additionally, these mutations were shown to occur in the strains of N315 type with higher probability rather than in those of MR108 type . The data suggest that one or more of these mutational sequences at least may have some influence on the expression of methicillin-resistance in clinical staphylococcal strains. Kansenshogaku Zasshi, 1994 Nov, 68(11), 1421 - 7 {A case of TSS complicated with SSSS in an adult with liver cirrhosis}; Toyota E et al.; This paper reports a case of TSS complicated with SSSS in an adult with liver cirrhosis . A 52-year-old male, heavy drinker, was referred to our clinic complaining lumbago and painful swelling of the right arm . The patient had peeling of the skin over the hips, knees and elbows with positive Nikolsky's sign . The patient was in a state of shock on admission . Pyrexia persisted for 4 days and finally the body temperature rose up to 39 degrees C . The laboratory studies revealed hypoxia, DIC and multiple organ failure, and these became progressively worse . He died 4 days after admission . According to the criteria, he was diagnosed as TSS, and TSST-1 was detected from his serum . Staphylococcus aureus, coagulase type V was cultured both from the blood and from the wound of his right middle finger . This isolated strain did not produce TSST-1 . The skin specimen at autopsy showed that the cleavage plane lied at the subcorneal region and close to the granular layer, with specific changes caused by exfoliative toxin . It was compatible to the exfoliation which was caused by exfoliative toxin produced from the S . aureus coagulase type V . The autopsy also revealed alcohol liver injury, liver cirrhosis and multiple organ failure due to shock state . SSSS is rare in adults, to our knowledge this is the first reported case of TTS complicated with SSSS. Nippon Ika Daigaku Zasshi, 1994 Nov, 61(6), 563 - 71 {Studies on the relationship between gastric acidity and the development of MRSA . Especially for the prevention of MRSA enterocolitis}; Suzuki S; Enterocolitis caused by methicillin-resistant Staphylococcus aureus (MRSA) has recently been recognized as one of the severe postoperative complications in surgery on the digestive organs . This disease often occurs in the early days after gastrointestinal operation, especially after gastrectomy . MRSA enterocolitis seems to occur when MRSA has first infected the naso-pharyngeal mucosa preoperatively, and then moved into the stomach, and subsequently proliferated in the higher pH gastric juices . The aim of this experiment was to reveal the relationship between the acidity of gastric juices and bacterial growth in the stomach during the pre and post operative period in an effort to prevent of MRSA enterocolitis . In vitro, MRSA was cultured for various periods at various pH values, and its proliferation was observed . MRSA did not grow in the culture at pH 1 at all, neither did it grow at pH 2 when cultured for more than 8 hours . This data shows the germicidal effect of high acidity in the stomach . Clinically, twenty patients with cancer in the digestive tract had the bacteria in their gastric juices examined in terms of acidity before and after operation . In cases with an increased pH level in the gastric juices after the operation . S . aureus including MRSA, was isolated frequently from the stomach . In vitro, incubation of MRSA with gastric juices collected from those cases showed no development of MRSA when the pH was below 3.98 . In order to prevent the onset of MRSA enterocolitis, the remnant stomachs of ten patients with stomach cancer were filled with hydrochloric acid lemonade just after operative reconstruction.(ABSTRACT TRUNCATED AT 250 WORDS) Chirurg, 1994 Nov, 65(11), 1023 - 7 {Arthroscopic distension irrigation in acute postoperative infection of the knee joint--long-term follow-up}; Riel KA et al.; Between 1983 and 1993, 26 patients, 12 patients from our clinic and 14 referred patients, had to be treated because of a septic knee joint following open ligament repair or subsequent arthroscopic surgery . Immediately after arthrocentesis for synovial-fluid analysis a distension-irrigation system was established under arthroscopic control as described by R.W . Jackson . Three tubes were placed into the knee joint cavity . One was used for inflow and the other two for outflow . Distension was performed by open inflow and closed outflow tubes each second hour for 10 to 15 min . Systemic antibiotics were administered . In 22 aspirated materials the cultures revealed bacteria: mostly staphylococcus aureus (65%) . In two cases in which irrigation of the knee-joint infection was performed later than the 10th postoperative day a relapse of the infection occurred and required a complete debridement and synovectomy . During follow-up period three patients underwent second look arthroscopy due to meniscus tear, rupture of an ACL reconstruction and synovial proliferation . 7 +/- 3 (2-13) years after treatment, 24 patients were re-examined, all had x-rays and 11 patients had a MRI of the knee joint additionally . 19 patients showed excellent or good results: no complaints, full ROM, no signs of osteoarthritis on x-rays and nearly normal participation in sports 4 out of 11 patients had signs of cartilage lesion and synovial proliferation . This retrospective study indicates that the distension-irrigation system is successful only in early acute cases of knee-joint infections of postoperative origins. Br Heart J, 1994 Nov, 72(5), 495 - 7 Persistent root abscess after emergency repair with an aortic homograft; Ritter M et al.; A fifty eight year old man with Marfan's syndrome and an aortic composite graft with a Bjork-Shiley mechanical prosthesis presented with a large aortic root abscess caused by Staphylococcus aureus endocarditis . Despite extensive surgical debridement and implantation of an aortic homograft as a composite graft, early postoperative transoesophageal echocardiography continued to demonstrate a large aortic root abscess and the patient died in a septic shock. J Dairy Sci, 1994 Nov, 77(11), 3354 - 64 Ecology of Staphylococcus aureus isolated from various sites on dairy farms; Roberson JR et al.; The purposes of this study were to identify sources of Staphylococcus aureus on dairies and to determine whether S . aureus colonization of heifer body sites increases the risk of S . aureus IMI at parturition . In herds with high (> 10%) or low (< 3%) prevalence of S . aureus IMI, S . aureus was isolated from heifer teat skin, heifer external orifices, housing, feedstuffs, humans, nonbovine animals, air, and equipment . Additionally, in herds with high prevalence, S . aureus was isolated from bedding, insects, and water . The predominant sources of S . aureus for both groups were other IMI and heifer body sites . Heifers with prepartum lacteal secretions with S . aureus were at greater risk of S . aureus IMI at parturition than were prepartum heifers with lacteal secretions that were negative for S . aureus . Heifers with teat skin colonized by S . aureus were 3.34 times more likely to have S . aureus IMI at parturition than were noncolonized heifers . Overall, 35% of 700 heifers were colonized with S . aureus on a body site at least once . Although colonizations of most body sites appeared to be transient, a few heifers were colonized on the same site for 1 yr . Persistently colonized heifers may represent the primary reservoirs of S . aureus for other heifers. J Dairy Sci, 1994 Nov, 77(11), 3347 - 53 Efficacy of intramuscular oxytetracycline as a dry cow treatment for Staphylococcus aureus mastitis; Erskine RJ et al.; To determine the efficacy of intramuscular oxytetracycline as a supplemental dry cow treatment for Staphylococcus aureus mastitis, 37 Holstein cows were randomly assigned to two treatment groups: intracisternal infusion with a commercial preparation of cephapirin benzathine at drying off (20 cows) and infusion with cephapirin benzathine at drying off and intramuscular oxytetracycline at 11 mg/kg once daily on d 7, 8, 9, and 10 after drying off (17 cows) . Milk samples collected 7, 14, 30, and 60 d after calving were plated for bacterial isolation within 24 h after collection and after 24 to 72 h of storage at -20 degrees C . Quarters were defined as infected if S . aureus was isolated from the fresh and frozen cultures from any one sample collected before drying off . An infected quarter was defined as cured if S . aureus was not isolated from the fresh or frozen culture from milk samples obtained following calving . The rate of cure by 30 d after calving for systemic oxytetracycline (in combination with cephapirin treatment) was 29.4% for infected quarters and 29.4% for infected cows, compared with 27.5 and 25.0%, respectively, for the cephapirin treatment only . Results including the culture at 60 d after calving were 21.2 and 22.5%, respectively, for combination therapy and cephapirin therapy only . Systemic oxytetracycline, in combination with intramammary dry cow treatment, did not improve the rate of cure for S . aureus mastitis. Microbiology, 1994 Nov, 140 ( Pt 11), 3139 - 44 Evidence for feedback (trans) regulation of, and two systems for, glycine betaine transport by Staphylococcus aureus; Stimeling KW et al.; Previous reports are in conflict as to the number of transport systems for glycine betaine in Staphylococcus aureus . Cells grown in complex medium exhibited a single transport system of moderate affinity . Cells grown in defined medium in the absence of glycine betaine showed a high affinity and a low affinity transport system . Cells grown in the presence of glycine betaine in the presence of osmotic stress in either complex or defined media accumulated large pools of internal glycine betaine . Smaller, but still significant, amounts of glycine betaine were accumulated by cells grown in its presence in either complex or defined media in the absence of osmotic stress . Cells grown in defined medium in the presence of glycine betaine in the presence or absence of osmotic stress showed lower rates of glycine betaine transport than cells grown in its absence . This suggests that glycine betaine transport is subject to feedback or trans inhibition by internal glycine betaine . This can explain the difference in observed kinetics in cells grown in complex or defined media, the high affinity system being predominantly inhibited in cells grown in complex medium. Microbiology, 1994 Nov, 140 ( Pt 11), 3131 - 8 Glycine betaine transport by Staphylococcus aureus: evidence for feedback regulation of the activity of the two transport systems; Pourkomailian B et al.; The regulation of glycine betaine accumulation by Staphylococcus aureus was investigated . The accumulation of glycine betaine was regulated by the osmotic pressure of the medium and the low affinity transport system played the major role in this regulation . Mutants were isolated that lack the low affinity, osmotically activated glycine betaine/proline transport system . Such mutants accumulated glycine betaine via the high affinity system but the glycine betaine pool was smaller and responded poorly to osmotic pressure changes . The regulation of glycine betaine transport has revealed that at the steady state net influx is reduced and that this is achieved by inhibition of both the low affinity and the high affinity transport systems . Cells pre-loaded with glycine betaine exhibited a reduced Vmax for both systems: the low affinity system was reduced in activity fivefold and the high affinity system was reduced 10-fold and became virtually undetectable . Although glycine betaine transport at the steady state is reduced, retention of the compatible solute is an active process since addition of an uncoupler provokes rapid release of the accumulated material . These data suggest that feedback regulation of the activity of the uptake systems is a major mechanism for controlling the level of compatible solute accumulation. Rev Med Suisse Romande, 1994 Nov, 114(11), 1035 - 43 {Computer alert and quality of care: application to the surveillance of hospital infections}; Safran E et al.; The Centre Informatique of Geneva University Hospital is developing, in the environment of its hospital information system, DIOGENE, a computerized alert system for surveillance of hospital infections . This hospital information system is based on an open distributed architecture and a relational database system, and covers many medical applications . This environment allows the development of alerts useful for detecting patients at risk . The alerts offer to clinicians a mean to control their efficacy in patient care . They are a new application of telematics for surveillance in clinical epidemiology, and are a tool for quality assurance . Two examples of alerts established for hospital infection control activities are presented . The first alert systematically detects all cases of patients colonized by or infected with methicillin-resistant Staphylococcus aureus (MRSA) . The second alert helps to organize prospective surveillance of bloodstream infections in order to identify some risk factors for infection and propose preventive measures. Photochem Photobiol, 1994 Nov, 60(5), 421 - 6 Spectroscopy and photosensitization of sapphyrins in solutions and biological membranes; Roitman L et al.; A spectroscopic and photophysical study of three new sapphyrin molecules is presented . The sapphyrin backbone that was derivatized to make them water soluble possesses an absorption band around 700 nm, a desired property for biological photosensitization . We studied the absorption and fluorescence spectra, from which evidence for aggregation in solvents of different polarities was obtained . The extent of aggregation is correlated with the nature of the attached moiety . The absolute quantum yields of singlet oxygen production were measured, with 1,3-diphenyl isobenzofuran as a model target, and were 0.13-0.18 in ethanol . The binding constants to liposomes and to cells were determined spectroscopically and were found to correspond to the hydrophobicities of the compounds, with an additional effect, ascribed to the sugar moiety, which was found in the case of one of the sapphyrins . The efficiency of photodamage to Staphylococcus aureus by sapphyrins and hematoporphyrin was equivalent, on the basis of cells killed per microgram of sensitizer in the incubation mixture. Mol Gen Mikrobiol Virusol, 1994 Nov-Dec, (6), 3 - 8 {Cloning and expression of the phosphatidylinositol-specific phospholipase C gene from Listeria monocytogenes}; Ermolaeva SA et al.; The gene for phosphatidylinositol-specific phospholipase C (PI-PLC) of Listeria monocytogenes has been cloned and shown to be expressed in Escherichia coli cells from own as well as from the lactose gene promoter . The recombinant plasmid has been constructed on the basis of pRIT2T vector and carries the hybrid gone . 3-end of which is a fragment of protein A gene of Staphylococcus aureus . 3-end is a gene for phospholipase plcA, both in the same reading frame . The resultant construction is shown to code in Escherichia coli cells for the hybrid recombinant protein A:Pl-PLC . Purified preparation of the hybrid protein and polyclonal rabbit antiserum to it were obtained . The obtained antiserum to the hybrid protein containing phospholipase as en C-end domain has been shown to react specifically to phospholipase in Escherichia coli recombinant strain harbouring the constructed recombinant plasmid as well as the one in the culture fluid of listeria. J Struct Biol, 1994 Nov-Dec, 113(3), 191 - 8 Imaging the spatial distribution of membrane receptors during neutrophil phagocytosis; Kindzelskii AL et al.; Optical microscopy and image processing have been employed to study the distribution of several cell surface receptors on living human neutrophils during opsonin-dependent and opsonin-independent phagocytosis . Receptors were labeled using fluorescein-, rhodamine-, or AMCA-conjugated F(ab')2 fragments of anti-Fc gamma RIIIB (CD16), anti-CR3 (CD11b/CD18), and anti-uPAR (urokinase-type plasminogen activator receptor) antibodies, intact phycoerythrin-labeled interleukin 8, and fluorescein- or rhodamine-labeled Con A (concanavalin A), Boc-PLPLP (tert-butyl-oxycarbonyl-Phe(D)-Leu-Phe(D)-Leu-Phe-OH), and N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys . Labeled neutrophils were observed during the phagocytosis of IgG-opsonized erythrocytes and nonopsonized latex beads, Escherichia coli, and Staphylococcus aureus . To quantitate receptor distribution, cells were divided into four quadrants with the first being the point of attachment and the fourth being opposite the point of attachment . Ligated formyl peptide receptors, and to a lesser extent CR3, accumulated at the sites of target internalization for all forms of phagocytosis examined . However, Fc gamma RIIIB, uPAR, IL-8, Con A, and the FPR antagonist FBoc-PLPLP were not polarized on cells during phagocytosis . These data suggest that agonist-labeled formyl peptide receptors may play a broader role in leukocyte function than previously suggested, including possible participation in phagocytosis. Antibiot Khimioter, 1994 Nov, 39(11), 29 - 36 {Additional differentiation of methicillin resistant strains of Staphylococcus aureus typed by the international phage bank}; Zyeva VS et al.; In the methicillin resistant strains of Staphylococcus aureus (MRSA) typed by the International Set phages the host specificity of the restriction-modification of the phage 85 DNA was determined, the finger printing of the cell DNA was carried out with using two probes and the lytic spectrum of the phages induced in them was studied . Four clones with different specificity of the restriction-modification system (rm89, rm108, rm121 and rm947) differing from that of strain PS 85 which is the host of phage 85 were detected . The strains belonging to the modification types m89, m108 and m121 contained prophages (within the respective groups) with similar lytic spectra when tested with the use of the PS strains of the International Collection and had cross antiphage immunity . Six phage variants were detected among the phages induced in the strains with the modification type m947 which could be indicative of the clone heterogeneity. Zentralbl Bakteriol, 1994 Nov, 281(4), 495 - 501 Role of antibodies against fibronectin-, collagen-binding proteins and alphatoxin in experimental Staphylococcus aureus peritonitis and septicaemia in neutropenic mice; Rozalska B et al.; We have investigated the protective role of hyperimmune rabbit IgG against two surface structures of Staphylococcus aureus, i.e . fibronectin-, and collagen-binding proteins as well as alpha-toxin in experimental peritonitis and septicaemia in neutropenic mice pretreated with cyclophosphamide . This treatment markedly decreased clearance of bacteria from mouse organs . With combined immunotherapy given passively bacteria were eradicated more efficiently for all animals sampled, comparative to controls. Intensive Care Med, 1994 Nov, 20 Suppl 4, S17 - 22 Anti-infective treatment in intensive care: the role of glycopeptides; Gruneberg RN et al.; Antibiotics are used in 80% of patients in the ICU, encouraging nosocomial infections with resistant organisms . If the antibiotic susceptibilities of the pathogen are known, a narrow-spectrum antibiotic is preferable to preserve the patient's resistance to colonization . However, treatment is often empirical and broad-spectrum combinations are commonly used . Gram-positive bacteraemia is associated with invasive monitoring or intravascular catheters . If the device cannot be removed easily, the glycopeptides are the only agents likely to be active against most strains of the commonest pathogen, the coagulase-negative staphylococcus . Long-stay patients are susceptible to infection with enterococci and methicillin-resistant Staphylococcus aureus, which are often resistant to all the usual agents other than glycopeptides . Vancomycin is long established, but is nephrotoxic, requires serum monitoring, must be administered as an infusion and can cause red man syndrome . Teicoplanin can be given as a single daily bolus without similar side-effects or monitoring . In deep-seated staphylococcal infection, the usual dose of teicoplanin is adequate if given in combination with other agents, but it may need to be doubled if used as monotherapy . Monitoring of the levels in the serum is helpful to ensure an adequate dose in patients with renal failure or in drug abusers, but is not needed to prevent toxicity. Infection, 1994 Nov-Dec, 22(6), 395 - 400 Infections related to the menstrual cycle . A study of five otherwise healthy women with recurrent abscesses and a review of the literature; Weischer M et al.; The purpose of the study was to investigate the in vitro bactericidal function of blood polymorphonuclear leucocytes (PMN) in various phases of the menstrual cycle from otherwise healthy women with recurrent cutaneous abscesses related to the premenstrual phase of the menstrual cycle compared with the bactericidal activity of PMN from healthy women with no inconveniences related to the menstrual cycle . The bactericidal activity against Staphylococcus aureus 502A was investigated and when possible against the patients' own strain . No variation in bactericidal activity was observed during the different phases of the menstrual cycle . PMN from five women with recurrent abscesses related to the premenstrual phase tended to kill fewer S . aureus 502A than PMN from three women in the control group . The literature of immunological defence mechanisms and the occurrence of infections related to the menstrual cycle is reviewed. Infection, 1994 Nov-Dec, 22(6), 390 - 4 Modern topical glucocorticoids and anti-infectives for superinfected atopic eczema: do prednicarbate and didecyldimethylammoniumchloride form a rational combination? Korting HC, Zienicke H, Braun-Falco O, Bork K, Milbradt R, Nolting S, Schopf E, Tronnier H. The addition of an anti-infective to a topical glucocorticoid preparation for superinfected atopic eczema is still controversial . To address this question in the context of the topical glucocorticoids of the non-halogenated double-ester type 0.25% prednicarbate cream was compared to the identical preparation incorporating the same amount of the disinfectant didecyldimethylammoniumchloride in patients suffering from atopic eczema carrying Staphylococcus aureus at a density of more than 10(6) colony-forming units per cm2 . One of the preparations was used twice daily over 5 days according to a random plan in a blind fashion . Thereafter treatment was based on either prednicarbate cream or the corresponding vehicle according to clinical needs . Clinical and microbiological evaluation were scheduled for days 0, 6 and 34 . Various clinical parameters were addressed individually as well as over all improvement using scores . A total of 143 patients was recruited . The patients of both groups improved rapidly with respect to clinical and microbiological findings . Essentially, there was no difference between the groups . Hence, the addition of an anti-infective to a topical prednicarbate preparation is not to be generally recommended. Stem Cells, 1994 Nov, 12(6), 626 - 37 Production of functional myeloid cells from CD34-selected hematopoietic progenitor cells using a clinically relevant ex vivo expansion system; Lill MC et al.; There is increasing clinical interest focused on ex vivo manipulation and expansion of hematopoietic cells . In this study, we demonstrate that a simple combination of growth factors can expand progenitors to yield functional myeloid cells . Furthermore, this system can produce mature, functionally competent cells in the absence of fetal bovine serum (FBS), which will enhance the clinical utility of this approach . Hematopoietic progenitor cells obtained from normal bone marrow and from leukapheresis products were studied . The mononuclear fraction was enriched for CD34 cells using the Ceprate CD34 biotin kit (CellPro #LC34-1 or LC34-2) . The selected cells were expanded for two weeks in Iscove's medium supplemented with 20% FBS and various combinations of interleukin-3 (IL-3), granulocyte colony-stimulating factor (G-CSF), stem cell factor (SCF) and interleukin -6 (IL-6) added either simultaneously or sequentially . The optimal combination of these factors identified for myeloid expansion was simultaneous addition of IL-3, SCF and G-CSF (at 50 ng/ml each), resulting in an average 773 +/- 133-fold expansion of nucleated cells (n = 5) . When corrected for the purity of CD34 cells in the starting population, the mean fold expansion with IL-3, SCF and G-CSF was 2,265 +/- 729 . A mean of 74.7 +/- 10.5% (n = 3) of the expanded cells was positive for CD11b; 86-91% (n = 2) of the cells were promyelocytes or more mature granulocytes . Functional assays demonstrated normal phagocytosis and intracellular killing of Staphylococcus aureus (S . aureus) by the expanded cell population . Studies performed using cells expanded in defined serum-free media demonstrated that fold expansion was decreased and that the cells produced were less mature and functionally less competent than cells expanded with FBS . The decreased expansion could be partially reversed, and the functionality almost completely restored by the addition of autologous plasma. Aten Primaria, 1994 Oct 31, 14(7), 892 - 4 {Prevalence of asymptomatic carriers of pathogenic germs in the nasopharynx . Comparison made between primary health care professionals and the population seeking care}; Chicote Feo R et al.; OBJECTIVE . To determine if the prevalence of non-symptomatic nasopharyngeal carriers of pathogenic germs is higher in health workers of primary care than in the general population . DESIGN . Cross-sectional, observational study . SETTING . Primary Care . Salamanca Urban Area . PARTICIPANTS . Seventy-six Health Centre workers from Salamanca and 152 individuals representing the general population attending the Health Centers were studied . People with pathology related to the study subject or with inmunitary problems were excluded . MEASUREMENTS AND MAIN RESULTS . A nasopharyngeal sample was carried out using a swab . The relative prevalence of carriers (sanitary workers vs general population) was 0.38 (c.i . 0.20-0.72) for Staphylococcus aureus and 0.56 (c.i . 0.34-0.93) for all pathogens . No significant differences were found according to sex . CONCLUSIONS . The prevalence of non-symptomatic nasopharyngeal carriers is higher in general population than in the group of primary care workers. Arch Intern Med, 1994 Oct 24, 154(20), 2330 - 5 Staphylococcus aureus endocarditis at a community teaching hospital, 1980 to 1991 . An analysis of 106 cases; Watanakunakorn C; BACKGROUND: The clinical diagnosis of infective endocarditis due to Staphylococcus aureus can be difficult, and many patients with this disease are only diagnosed post mortem . There are few published reports of large series of patients with S aureus endocarditis and none from a community hospital . I reviewed the clinical and laboratory findings of a large number of patients with S aureus endocarditis in a community hospital . METHODS: I reviewed medical records identified through consultation records, International Classification of Diseases, Ninth Edition codes, and autopsy records of patients who fulfilled the criteria for the diagnosis of S aureus endocarditis during 1980 to 1991 . RESULTS: During the 12-year period, there were 106 cases, for a prevalence of 0.34 per 1000 admissions . Ninety-three (87.7%) of these patients were seen by me . The patients' ages ranged from 12 to 83 years (median, 61 years) . Eighteen cases were nosocomial (15 were associated with intravascular catheters) . Twenty-one patients were injecting drug users . Severe back pain was the chief complaint in nine patients . Twenty-seven patients had no heart murmur at the time of diagnosis . The overall mortality was 25.5% . CONCLUSIONS: Age 60 years or older, female gender, community-acquired infection, absence of heart murmur, presence of congestive heart failure, or central nervous system involvement was associated with higher mortality . Tricuspid valve endocarditis alone was associated with lower mortality. Biochem J, 1994 Oct 15, 303 ( Pt 2), 633 - 8 Activation of the glycosyl-phosphatidylinositol-anchored membrane dipeptidase upon release from pig kidney membranes by phospholipase C; Brewis IA et al.; Incubation of pig kidney microvillar membranes with Bacillus thuringiensis or Staphylococcus aureus phosphatidylinositol-specific phospholipase C (PI-PLC) resulted in the release of a number of glycosyl-phosphatidylinositol (GPI)-anchored hydrolases, including alkaline phosphatase (EC 3.1.3.1), amino-peptidase P (EC 3.4.11.9), membrane dipeptidase (EC 3.4.13.19), 5'-nucleotidase (EC 3.1.3.5) and trehalase (EC 3.2.1.28) . Of these five ectoenzymes only for membrane dipeptidase was there a significant (approx . 100%) increase in enzymic activity upon release from the membrane . Maximal activation occurred at a PI-PLC concentration 10-fold less than that required for maximal release . In contrast solubilization of the membranes with n-octyl beta-D-glucopyranoside had no effect on the enzymic activity of membrane dipeptidase . A competitive e.l.i.s.a . with a polyclonal antiserum to membrane dipeptidase indicated that the increase in enzymic activity was not due to an increase in the amount of membrane dipeptidase protein . Although PI-PLC cleaved the GPI anchor of the affinity-purified amphipathic form of pig membrane dipeptidase there was no concurrent increase in enzymic activity . In the absence of PI-PLC, membrane dipeptidase in the microvillar membranes hydrolysed Gly-D-Phe with a Km of 0.77 mM and a Vmax . of 602 nmol/min per mg of protein . However, in the presence of a concentration of PI-PLC which caused maximal release from the membrane and maximal activation of membrane dipeptidase the Km was decreased to 0.07 mM while the Vmax . remained essentially unchanged at 624 nmol/min per mg of protein . Overall these results suggest that cleavage by PI-PLC of the GPI anchor on membrane dipeptidase may relax conformational constraints on the active site of the enzyme which exist when it is anchored in the lipid bilayer, thus resulting in an increase in the affinity of the active site for substrate. Biochem J, 1994 Oct 15, 303 ( Pt 2), 517 - 25 The intracellular distribution of inositol polyphosphates in HL60 promyeloid cells; Stuart JA et al.; 1 . HL60 promyeloid cells contain high intracellular concentrations of inositol polyphosphates, notably inositol 1,3,4,5,6-pentakisphosphate (InsP5) and inositol hexakisphosphate (InsP6) . To determine their intracellular location(s), we studied the release of inositol (poly)phosphates, of ATP, and of cytosolic and granule-enclosed enzymes from cells permeabilized by four different methods . 2 . When cells were treated with digitonin, all of the inositol phosphates were released in parallel with the cytosolic constituents . Most of the InsP5 and InsP6 was release