Staphylococcus

Acta Microbiol Pol, 1992, 41(3-4), 145 - 50
Staphylococcal L-asparaginase: catabolic repression of synthesis; Rozalska M et al.; The regulation mechanism of production of staphylococcal L-asparaginase was investigated . The role of cAMP in regulation of its synthesis was confirmed . Production of L-asparaginase from S . aureus NCTC 4135 was inhibited by all carbon sources, mono- and disaccharides added to the growth medium . The strongest inhibition was caused by saccharose and maltose, whereas weaker by galactose, lactose, mannitol and mannose . It was found that exogenous cAMP in the presence of carbon sources stimulated synthesis of the investigated enzyme.

Ann Trop Paediatr, 1992, 12(4), 451 - 3
Pyogenic liver abscess causing acute Budd-Chiari syndrome; Mehrotra G et al.; Acute Budd-Chiari syndrome caused by compression of the inferior vena cava by a space-occupying lesion in the liver is rare in children . We report a case where the compression was due to a large staphylococcal abscess in the right lobe of the liver . A high index of suspicion in such a case ensures early intervention and saves life.

Microbios, 1992, 71(288-289), 203 - 15
Detection and purification of a potential precursor protein or a prohaemolysin produced by Staphylococcus haemolyticus; Sevigny G et al.; The haemolytic substance H7 produced by Staphylococcus haemolyticus is composed of three peptides out of 44 amino acid residues each having a structure resembling a signal peptide . The cytoplasmic fraction for the presence of a protein precursor containing this signal sequence was investigated . Specific rabbit IgGs to substance H7 were isolated by affinity chromatography on Sepharose-4B-H7 . These anti-H7 IgGs recognized mainly a 51 kD protein in the cytoplasmic fraction of S . haemolyticus from 2, 4, 6, 8, 10 and 23 h cultures . These results support the idea that the 51 kD protein could be either a prohaemolysin or else the precursor of a protein of unknown function with a signal sequence showing homology with the haemolytic peptides . After affinity chromatography on Sepharose-4B-anti-H7, the 51 kD protein was shown associated with an RNA-protein complex composed of four or five proteins and an RNA estimated at 300 nt . This complex could be associated with the machinery of protein secretion . The 51 kD protein was finally purified to homogeneity by HPLC on a Protein Pak DEAE-5PW column in the presence of 5 M urea.

J Immunol, 1991 Dec 15, 147(12), 4082 - 5
A recombinant C-terminal fragment of staphylococcal enterotoxin A binds to human MHC class II products but does not activate T cells; Hedlund G et al.; Binding of staphylococcal enterotoxin A (SEA) to MHC class II encoded proteins is a prerequisite for its subsequent activation of a large fraction of T lymphocytes through interaction with variable segments of the TCR-beta chain . We cloned SEA in Escherichia coli and produced four recombinant fragments covering both the N- and C-terminal regions . These fragments were used to analyze the interaction between SEA and the human MHC class II products . A C-terminal fragment of SEA, representing amino acids 107-233 bound to HLA-DR and HLA-DP but did not activate T cells . The three other fragments (amino acids 1-125, 1-179 and 126-233) neither bound to MHC class II Ag nor activated T cells . SEA apparently bind to HLA-DR and HLA-DP through its C-terminal part, whereas T cell activation is dependent on additional parts of the protein.

Biochem J, 1991 Dec 15, 280 ( Pt 3), 809 - 11
Identification by protein microsequencing of a proteinase-V8-cleavage site in a folding intermediate of chick muscle creatine kinase; Morris GE et al.; We have identified by protein microsequencing a glutamic acid residue (Glu-166) in a folding intermediate of chick muscle creatine kinase that is very sensitive to cleavage by staphylococcal proteinase V8 . Most other glutamic acid residues, including Glu-168, are already partly protected from proteolytic attack at this stage . After the final stages of protein refolding, when enzyme activity is recovered, Glu-166 is also resistant to proteolysis.

J Long Term Eff Med Implants, 1992, 1(4), 321 - 8
The inhibition of bacterial adhesion to a tobramycin-impregnated polymethylmethacrylate substratum; Oga M et al.; Tobramycin sulfate powder (1.2 g) was mixed with Palacos polymethylmethacrylate (PMMA) bone cement (40 g) to produce 100 discs containing 5.9 mg tobramycin per disc . These discs were used to evaluate the inhibition of bacterial adhesion to an antibiotic-laden biomaterial . Tobramycin-impregnated PMMA discs and control discs containing no tobramycin were exposed in vitro to Staphylococcus epidermidis . Colonization was quantitated using plate count techniques and electron microscopy . Tobramycin-impregnated surfaces reduced adhesive bacteria colonization by 1 log relative to control discs . These observations suggest that tobramycin-impregnated PMMA may not be significantly effective in preventing colonization of the biomaterial substratum and PMMA may be a poor choice as a drug delivery vehicle in biomaterial and compromised tissue-centered infections.

Eur J Biochem, 1991 Dec 5, 202(2), 421 - 9
Pericytes of the brain microvasculature express gamma-glutamyl transpeptidase; Frey A et al.; The expression of gamma-glutamyl transpeptidase (GGT) is a specific property of the brain capillary endothelium that constitutes the blood-brain barrier . We report here the detection of GGT, not only in endothelial cells, but also in pericytes, demonstrating that a brain capillary-specific pericyte population exists . We raised antibodies to GGT using a porcine brain microvessel GGT-protein-A (staphylococcal protein A) fusion protein as antigen which was expressed in Escherichia coli . The immunohistochemical analysis of the subcapillary distribution of GGT in porcine brain cortex and cerebellum sections by both light and electron microscopy revealed the expression of GGT in the capillary-adjacent pericytes in addition to the GGT-positive endothelial layer . We confirmed these data for cultured porcine brain microvascular endothelial cells and pericytes . GGT immunofluorescence could be detected in both cell types in culture . Endothelial cells exhibited a weak staining, whereas pericytes were strongly positive for GGT . Due to the high phagocytotic activity of pericytes and their location on the abluminal surface of the microvessels, we propose a possible protective or detoxifying function of GGT in cerebrovascular pericytes.

Circulation, 1991 Dec, 84(6), 2539 - 46
Protection against endocarditis due to Staphylococcus epidermidis by immunization with capsular polysaccharide/adhesin; Takeda S et al.; BACKGROUND . Staphylococcus epidermidis is the principal pathogen in prosthetic valve endocarditis . The capsular polysaccharide adhesin (PS/A) has been shown to mediate attachment of bacteria to medical devices . In this study, we investigated the efficacy of active and passive immunization against PS/A in preventing S . epidermidis endocarditis in a rabbit model . METHODS AND RESULTS . Aortic valve vegetations were produced by inserting a Teflon catheter into the left ventricle through the right carotid artery . Bacteremia and endocarditis were then established by implanting in the left jugular vein a catheter that was attached to an osmotic pump and contaminated with S . epidermidis strain RP62A . During a 3-week study period, of 64 blood cultures taken every second or third day from six nonimmune rabbits, 54 (84%) yielded strain RP62A . In rabbits actively immunized with PS/A, eight of 60 blood cultures (13%) were positive (odds ratio 5.0, 95% CI, 2.0-12.3, p = 0.005) . At death, all six nonimmune rabbits had infected vegetations that yielded 10(6)-10(11) colony-forming units (cfu)/g of vegetation, whereas only one PS/A-immunized rabbit had an infected vegetation . Immunization protocols designed to elicit antibody to teichoic acid but not to PS/A afforded no protection against bacteremia or endocarditis . Infusion of monoclonal antibody to PS/A through a catheter in the right jugular vein provided a level of protection against both bacteremia and endocarditis comparable to that produced by active immunization . In vitro, antibody against PS/A was opsonic for S . epidermidis . CONCLUSIONS . Immunoprophylaxis targeted at staphylococcal PS/A is a promising new approach to the prevention of prosthetic valve endocarditis.

J Immunol, 1991 Dec 1, 147(11), 3876 - 81
Staphylococcal toxins bind to different sites on HLA-DR; Chintagumpala MM et al.; Staphylococcal enterotoxins (SE) and toxic shock syndrome toxin 1 (TSST-1) bind to MHC class II molecules and the toxin-class II complexes induce proliferation of T cells bearing specific V beta sequences . We have previously reported that these toxins display varying binding affinities for HLA-DR1 . We now investigated whether these differences simply reflected differences in binding affinity for a single class II binding site or, at least in part, the engagement of different binding sites on the HLA-DR complex . Through competitive binding studies we show that SEB and TSST-1, which are not closely related by their amino acid sequences, bind to two different sites on HLA-DR . Both of these sites are also occupied by staphylococcal enterotoxin A (SEA), enterotoxin D (SED), and enterotoxin E (SEE) which exhibit more than 70% amino acid sequence homology . SEB and TSST-1 failed to inhibit SEA binding to HLA-DR . These studies suggest that there may be three distinct, although perhaps overlapping, binding sites on HLA-DR for these toxins . Further, although SED and SEE are similar to SEA in structure, and appear to bind the same sites on HLA-DR as SEA, they displayed significantly lower binding affinities . T cell proliferative responses to SED required a higher concentration of the toxin than SEA, probably reflecting its lower binding affinity . SEE, however, elicited T cell responses at very low concentrations, similar to SEA, despite its much lower binding affinity . Therefore, although the affinities of these toxins to MHC class II molecules appear to significantly influence the T cell responses, the effective recognition of the toxin-class II complex by the TCR may also contribute to such responses.

Immunology, 1991 Dec, 74(4), 638 - 44
The stage of negative selection in tolerance induction in neonatal mice; Ando T et al.; In flow microfluorometry analysis of thymus and lymph node cells of C57BL/6(I-E-,Mlsb) mice rendered neonatally tolerant to (C57BL/6 x AKR/J)F1 (I-E+,Mlsb/a) lymphoid cells, both CD4+ and CD8+ cells showed a striking reduction in the number of V beta 6+ cells capable of recognizing Mlsa in the context of major histocompatibility complex (MHC) class II molecules, indicating that clonal deletion of V beta 6+ cells by Mlsa antigen occurs just at a stage of immature V beta 6lowCD4+CD8+ thymocytes . On the other hand, the number of V beta 11+ cells capable of recognizing I-E was markedly reduced in CD4+ cells, but CD8+ cells showed only partial (20%) reduction of such a population . The clonal deletion of V beta 11+ cells by I-E may begin at the transitional stage from V beta 11lowCD4+CD8+ to V beta 11highCD4+CD8- single-position cells, and V beta 11lowCD4+CD8+ cells differentiating to V beta 11highCD4-CD8+ cells seem to be resistant to clonal deletion . V beta 11+ T cells are also stimulated by staphylococcal enterotoxin A (SEA) irrespective of expression of CD4 or CD8 . Nearly all of both V beta 11+CD4+ and V beta 11+CD8+ lymph node T cells were deleted by the injection of SEA every other day from birth . In their thymi, both V beta 11+CD4+CD8- and V beta 11+CD4-CD8+ single-positive thymocytes were deleted, and the proportion of V beta 11low thymocytes was lower than that of normal mice . The clonal deletion of V beta 11+ T cells by SEA injection occurs at a stage of immature V beta 11lowCD4+CD8+ double-positive thymocytes, resulting in deletion of both V beta 11+CD4+ and V beta 11+CD8+ T cells.

J Protein Chem, 1991 Dec, 10(6), 669 - 83
Biosynthetic antibody binding sites: development of a single-chain Fv model based on antidinitrophenol IgA myeloma MOPC 315; McCartney JE et al.; The functional antigen binding region of antidinitrophenol mouse IgA myeloma MOPC 315 has been produced as a single-chain Fv (sFv) protein in E . coli . Recombinant 315 proteins included sFv alone, a bifunctional fusion protein with amino-terminal fragment B (FB) of staphylococcal protein A, and a two-chain 315 Fv fragment . Successful refolding of the 315 sFv required formation of disulfide bonds while the polypeptide was in a denatured state, as previously observed for the parent Fv fragment . Affinity-purified recombinant 315 proteins showed full recovery of specific activity, with values for Ka,app of 1.5 to 2.2 x 10(6) M-1, equivalent to the parent 315 Fv fragment . As observed for natural 315 Fv, the sFv region of active FB-sFv315 fusion protein was resistant to pepsin treatment, whereas inactive protein was readily degraded . These experiments will allow the application of protein engineering to the 315 single-chain Fv; such studies can advance structure-function studies of antibody combining sites and lead to an improved understanding of single-chain Fv proteins.

Rev Esp Fisiol, 1991 Dec, 47(4), 201 - 8
Influence of blood proteins in the in vitro adhesion of Staphylococcus epidermidis to teflon, polycarbonate, polyethylene and bovine pericardium; Carballo J et al.; The influence of human plasma proteins (fibrinogen, albumin and fibronectin) on the adherence of Staphylococcus epidermis to teflon, polyethylene, polycarbonate and bovine pericardium was studied in an in vitro quantitative assay by scintillation counting . Bacterial adhesion was generally reduced by the presence of protein during the adherence assay except in the case of bovine pericardium, in which adherence remained almost unaffected . The effect of these plasma proteins on bacterial surface properties resulted in strong increases of surface charge as measured by ion-exchange chromatography and with no effect on hydrophobicity, estimated as contact angles . Adherence was not found to be correlated with these two properties, suggesting that bacteria-surface interactions must not be simplified to the influence of interfacial forces.

Eur J Clin Microbiol Infect Dis, 1991 Dec, 10(12), 1007 - 12
In vitro selection of resistance to vancomycin in bloodstream isolates of Staphylococcus haemolyticus and Staphylococcus epidermidis; Herwaldt L et al.; The purpose of the study was to determine whether vancomycin-resistant strains of Staphylococcus haemolyticus could be selected regardless of the initial MIC of vancomycin . Twenty-one bloodstream isolates of Staphylococcus haemolyticus were studied by broth and agar selection methods . The broth method selected strains for which MICs of vancomycin ranged from 4 to 32 micrograms/ml and MBCs from 16 to greater than 128 micrograms/ml . The agar method selected strains for which MICs ranged from 8 to 32 micrograms/ml and MBCs from 8 to greater than 128 micrograms/ml . For comparison, seven strains of Staphylococcus epidermidis were evaluated by the agar selection method . Final MICs of vancomycin ranged from 8 to 16 micrograms/ml; MBCs ranged from 16 to 64 micrograms/ml . Clearly, in vitro exposure to vancomycin can select strains of Staphylococcus haemolyticus and Staphylococcus epidermidis for which MIC values are beyond the susceptible breakpoint.

J Rheumatol, 1991 Dec, 18(12), 1841 - 4
A prospective analysis of patients with rheumatic diseases attending referral hospitals in Harare, Zimbabwe; Stein CM et al.; In a 10-month prospective study a research assistant identified 411 patients with rheumatic disease at the 2 referral hospitals in Harare . Rheumatic disease accounted for less than 1% of hospital admissions . Rheumatoid arthritis, the commonest condition, accounted for 18% of patients, many of whom had impaired functional capacity . Septic arthritis (16%) was common in younger patients, often affecting the hip or knee and often associated with other complications of disseminated staphylococcal infection . Osteoarthritis (9%), rheumatic fever (7%), gout (6%), human immunodeficiency virus associated musculoskeletal problems (6%) and systemic lupus erythematosus (5%) were relatively common while the spondyloarthropathies occurred less frequently . The spectrum of rheumatic disease seen in teaching hospitals in Harare, although significantly different from that seen in Europe and North America, approximates the pattern seen in developed countries more closely than previous studies from Africa would suggest.

Int J Food Microbiol, 1991 Dec, 14(3-4), 325 - 31
Feasibility of a reference material for staphylococcal enterotoxin A; Notermans S et al.; A reference material for staphylococcal enterotoxin A (SEA), was produced by spray-drying the toxin in milk . With this procedure the SEA was distributed homogeneously in the material . For ease of handling the reference material was encased in gelatin capsules, each containing 405 ng of SEA . Simply dissolving the milk powder in distilled water resulted in a 100% recovery of the SEA present . The reference material would appear suitable for testing laboratory performance, comparison of detection methods and to validation of extraction procedures.

Int J Food Microbiol, 1991 Dec, 14(3-4), 305 - 12
Comparison of four kits for the detection of staphylococcal enterotoxin in foods from outbreaks of food poisoning; Wieneke AA; Four commercial kits, three based on sandwich ELISA techniques and one on latex agglutination were compared for the detection of staphylococcal enterotoxins in foods from outbreaks of food poisoning . Enterotoxin was detected in 14 of 18 foods with the Swiss SET-EIA and in 9 or 10 with the Unipath SET-RPLA and two ELISAs from Transia . The advantages and disadvantages of the four methods are discussed.

Zentralbl Bakteriol, 1991 Dec, 276(1), 86 - 93
Staphylococcal lipoteichoic acid exerts growth factor-like activity towards human and murine cells; Ohshima Y et al.; Lipoteichoic acid (LTA) was extracted from Staphylococcus saprophyticus strain S1 and tested for the capacity to induce hematopoietic and lymphatic cell proliferation . As compared to nontreated cells, the number of human bone marrow cells significantly increased in the presence of low LTA concentrations . Optimal growth was observed on the fifth day of in vitro incubation . After exposure to LTA, the lymphocyte proliferation rate also increased in a dose and time dependent manner . On the other hand, human epithelial cells and fibroblasts did not show enhanced growth activities in the presence of LTA.

Zentralbl Bakteriol, 1991 Dec, 276(1), 36 - 45
Evidence for degradation of synthetic polyurethanes by Staphylococcus epidermidis; Jansen B et al.; The survival of Staphylococcus epidermidis strain KH 11 in the presence of synthetic high molecular polyurethanes was prolonged in comparison to control experiments performed in the absence of any nutrients . Investigations of the bacteria after contact with the polymers revealed changes in their surface properties and metabolism, in particular a marked induction of urease activity . ESCA (Electron Spectroscopy for Chemical Analysis) measurements detected a decrease in elementary nitrogen in the polyurethane surfaces after incubation with the bacteria . The alterations observed indicate an urease-induced degradation of synthetic polymers by Staphylococcus epidermidis KH 11.

Zh Mikrobiol Epidemiol Immunobiol, 1991 Dec, (12), 42 - 5
{The reactogenicity and efficacy of a dried cell-free staphylococcal vaccine in treating chronic bronchitis}; Egorova NB et al.; The data obtained in the controlled trial of the immunotherapy of chronic bronchitis (CB) patients with lyophilized acellular staphylococcal vaccine developed at the Mechnikov Research Institute for Vaccines and Sera (USSR Acad . Med . Sci.), Moscow, are presented . The patients were divided at random into two groups; of these, one group received the vaccine and the other, placebo (0.9% sodium chloride solution) . The preparations were injected subcutaneously simultaneously with traditional treatment . The vaccine was well tolerated by the patients . In the blood of the patients receiving the vaccine, in contrast to those receiving placebo, a significant increase in the level of specific antibodies determined in the passive hemagglutination test and IgG was noted . When following the remote consequences of the disease for a year after the course of immunotherapy, a significant decrease in the occurrence of aggravations, an increase in the duration of remission and a decrease in the duration of sick leaves were registered . These data indicate that the immunotherapy of CB with lyophilized acellular staphylococcal vaccine is a promising method of preventing relapses.

Klin Med (Mosk), 1991 Dec, 69(12), 53 - 5
{Evaluation of diagnostic indices of reactivity during treatment of patients with chronic bronchitis and bronchiectatic disease}; Linevskii IuV et al.; Evaluation of neutrophils conditions by glycogen, enzymatic activity (acid and alkaline phosphatases, peroxidase), lymphocyte blast-transformation reaction stimulated by staphylococcal antigen in 113 patients with bronchial pathology on treatment suggested the conclusion of their value in verification of the disease course correction of on-going treatment and definition of its effect . The methods acquire additional significance in objective assessment of the process activity in obscure clinical manifestations and lack of laboratory evidence for aggravation.

Br J Ophthalmol, 1991 Dec, 75(12), 710 - 2
Posterior lens capsule abscess due to Propionibacterium acnes and Staphylococcus epidermidis following extracapsular cataract extraction; Manners RM et al.; A case of posterior lens capsular abscess occurring many months after an extracapsular cataract extraction is presented . This was caused by a mixed infection involving Propionibacterium acnes and Staphylococcus epidermidis . The significance of Staph epidermidis after such a long postoperative period is uncertain, but the case shows features typical of secondary endophthalmitis due to P acnes, including a long delay in onset and a grumbling course not brought under control by medical treatment . It supports the theory that the nidus of infection is localised in the posterior lens capsule by showing development of a visible capsular abscess with associated vitreous involvement . The subsequent removal of the capsule and vitreous, despite leaving the intraocular lens in place, led to complete resolution of the inflammation . Both organisms have previously been found to be sequestered in the posterior lens capsule by histological and microbiological examination of excised capsular specimens . It is important to consider them as possible causative agents in the formation of a postoperative capsular abscess.

Kidney Int, 1991 Dec, 40(6), 1160 - 5
Simultaneous catheter replacement and removal in refractory peritoneal dialysis infections; Swartz R et al.; The present report describes more than nine years of experience with simultaneous removal and replacement of the chronic peritoneal dialysis (PD) catheter in treating refractory mechanical and infectious complications . Simultaneous catheter replacement and removal not only succeeded in 22 of 23 cases with non-infectious mechanical complications, allowing uninterrupted PD in all but three, but also succeeded in 30 (83%) of 36 additional cases with persisting or recurring infection . Simultaneous procedures failed in six (17%) of 36 infectious cases, due to persistent infection in two cases and to procedure complications in four cases . The clinical characteristics of the 30 successful infectious cases were compared to the characteristics of both the six failures, as well as 29 additional infectious cases in which the peritoneal catheter was removed but was not replaced because of a variety of serious complications (such as pancreatitis, abscess, sepsis, or fluid overload) . At the time of simultaneous catheter replacement in successful cases, temporary control of active peritoneal inflammation was achieved more frequently (80% vs . 31%, P less than 0.001) with a shorter interval of antibiotic coverage (6.4 +/- 0.9 vs . 14.7 +/- 1.3 days, P less than 0.001) . In addition, the successful cases were characterized by significantly more Staphylococcal infections (70% vs . 26%, P less than 0.001) and significantly fewer Pseudomonal or fungal infections (6% vs . 59%, P less than 0.001), although successful cases included some non-Pseudomonal, non-enteric gram negative infections (23%).(ABSTRACT TRUNCATED AT 250 WORDS)

Eur J Vasc Surg, 1991 Dec, 5(6), 627 - 32
The clinical use of an antibiotic-bonded graft; Strachan CJ et al.; Attempts to produce an antibiotic-bonded prosthesis have failed owing to poor binding, drug toxicity or inadequate antimicrobial activity of the antibiotic particularly against Staphylococcus epidermidis . Rifampicin, with an ideal spectrum, but untested against slime-forming S . epidermidis has recently been shown to bind with carboxyl groups on gelatin-sealed Dacron . We therefore investigated rifampicin activity against 30 slime-forming adherent S . epidermidis colonies, isolated from 40 consecutive aortic graft recipients, and compared it with their methicillin, gentamicin, cefuroxime, tetracycline and vancomycin resistance patterns . The S . epidermidis colonies were highly sensitive to very low levels of rifampicin . Rifampicin was then bonded to gelatin-sealed Dacron aortic prostheses which were inserted in four patients at high risk of developing subsequent infection.

Br J Rheumatol, 1991 Dec, 30(6), 464 - 7
A preliminary study of magnetic resonance relaxation times (T1 and T2) in inflammatory and degenerative synovial fluids; Bellamy N et al.; Multiple synovial fluid samples from 21 patients were analysed using standard synovial analysis techniques and by nuclear magnetic resonance spectroscopy . Significant negative correlations were noted between both T1 (P less than 0.01) and T2 (P less than 0.0006) relaxation times and synovial fluid total protein . No differences in T1 or T2 relaxation times were noted in synovial fluid between 16 patients with inflammatory forms of arthritis and five patients with degenerative arthritis . In a single rheumatoid arthritis patient with concurrent staphylococcal arthritis, T1 and T2 relaxation times did not vary between the active phase and the recovery phase . The lack of any significant differences in the measured relaxation times as a function of joint condition suggest that in vivo magnetic resonance measurements of T1 or T2 for joint analysis may not reveal information of either a diagnostic or pathophysiological nature.

J Exp Med, 1991 Dec 1, 174(6), 1431 - 7
Reduction of lupus nephritis in MRL/lpr mice by a bacterial superantigen treatment; Kim C et al.; The effects of biweekly intravenous injections of Staphylococcus Enterotoxin B (SEB) into autoimmune MRL-lpr/lpr (MRL/lpr) mice were investigated . Rather than causing the expansion of V beta 8+ T cells, SEB administration resulted in the reduction V beta 8+, CD4-CD8- "double-negative" (DN) T cells . This was shown by FACS analysis as this putative pathogenic population was diminished in both spleen and lymph node . The symptoms of systemic lupus erythematosus (SLE) in MRL/lpr, which include high titers of anti-DNA antibodies and circulating immune complexes and proteinuria, were reduced in SEB-treated mice in a dose-dependent manner . The clinical parameters of SLE in MRL/lpr, which include lymph node hyperplasia and necrotic vasculitis, were suppressed in 50-micrograms SEB-treated mice . T cells bearing V beta 6 T cell receptor, which does not interact with SEB, were not reduced with SEB administration . Thus, disease suppression was associated with a specific reduction in the number of V beta 8+, DN T cells . These results implicate a possible therapeutic role of superantigen-based immunotherapy in V beta-restricted, T cell-dominated clinical syndromes.

J Lab Clin Med, 1991 Dec, 118(6), 570 - 5
The primary structure of serum amyloid A protein in the rabbit: comparison with serum amyloid A proteins in other species; Liepnieks JJ et al.; Rabbit serum amyloid A (SAA) protein was isolated from acute-phase serum by ultracentrifugation, molecular seive chromatography, and ion-exchange chromatography . The complete amino acid sequence of the protein was established by sequence analysis of peptides derived from trypsin and Staphylococcus proteinase digestion of the protein . The molecule consisted of 104 amino acids and had an amino terminus that was blocked by pyrrolidonecarboxylic acid . Heterogeneity was not observed at any residue, which suggests that the material sequenced consisted of a single serum amyloid A species . The protein is highly homologous to serum amyloid A from humans and other animals, particularly in the middle portion of the molecule (positions 33 to 63), which suggests that this region may be important in its function . This highly conserved region may also contain the determinants for amyloid formation.

J Perinatol, 1991 Dec, 11(4), 319 - 25
Acute phase reactants in neonatal bacterial infection; Pourcyrous M et al.; The C-reactive protein (CRP) level was evaluated in 142 infants requiring investigation for suspected infection . After excluding two neonates because of incomplete data, there remained 140 neonates, of whom 16 had septicemia . Fifteen of 16 had increased CRP levels . The CRP value was not elevated in any baby (n = 5) who had positive blood cultures for Staphylococcus epidermidis, all of whom had an uneventful clinical course . The CRP level was elevated in all six babies with meconium-aspiration syndrome, but was normal in five infants whose viral cultures were positive . Ninety-nine percent of uninfected babies had normal CRP values . Overall, CRP was a valuable test for diagnostic confirmation of bacterial infection . Elevated CRP level was always accompanied by at least one abnormality in the other tests performed . Although the study was not intended to predict clinical onset of bacterial disease, our results suggest that the CRP level, because of a high negative predictive value, may be useful in ruling out bacterial infection.

J Clin Microbiol, 1991 Dec, 29(12), 2768 - 73
Molecular relatedness of Staphylococcus epidermidis isolates obtained during a platelet transfusion-associated episode of sepsis; Shayegani M et al.; Staphylococcus epidermidis was isolated from the blood of a 25-year-old pregnant woman following the administration of eight units of platelets . She had developed chills and a fever of 41.4 degrees C soon after the transfusions were completed . S . epidermidis was also obtained from all eight platelet units, as well as from the packed-erythrocyte unit associated with the first unit of platelets . The isolation of the same organism from these epidemiologically related sources provided us with the opportunity to phenotypically and genetically characterize the isolates . Several typing methods, including four molecular techniques, were used to increase our chances of finding any differences between the isolates under investigation . Phenotypic analyses demonstrated that S . epidermidis isolates from the patient, platelet units, and erythrocyte unit reacted in exactly the same manner in 15 biochemical tests, exhibited slime production, and had the same antibiotic susceptibility pattern . Genetic analyses, which included plasmid profiles, plasmid cross-hybridization, field inversion gel electrophoresis, and ribotyping, substantiated the relationships between the S . epidermidis isolates from the patient, platelet units, and erythrocyte unit . Eight S . epidermidis control strains unrelated to the case were found to differ significantly from the platelet-related strain.

J Biol Chem, 1991 Nov 25, 266(33), 22569 - 81
The beta-glucan synthase from Lolium multiflorum . Detergent solubilization, purification using monoclonal antibodies, and photoaffinity labeling with a novel photoreactive pyrimidine analogue of uridine 5'-diphosphoglucose; Meikle PJ et al.; The membrane-bound beta-glucan synthase from Italian ryegrass (Lolium multiflorum L.) endosperm cells has been solubilized by both non-ionic and zwitterionic detergents . A complex relationship exists between the ratio of (1----3)-, (1----4)-, and (1----3, 1----4)-beta-glucan products of the solubilized enzyme, the cations present, and the concentration of the uridine 5'-diphosphoglucose substrate . Monoclonal antibodies directed against the beta-glucan synthase complex were generated by immunization of mice with an unfractionated microsomal reparation . Hybridoma cell lines were screened using a combination of indirect enzyme-linked immunosorbent assay followed by an enzyme-capture assay . The purified monoclonal antibodies were used with Pan-sorbin (stablized protein A-bearing staphylococcal cells) to immunoprecipitate an active beta-glucan synthase complex which had been solubilized from a microsomal preparation with 0.6% CHAPS . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the immunoprecipitated synthase complex revealed four major polypeptides of apparent molecular mass 30, 31, 54, and 58 kDa together with several minor components . The immunoprecipitated beta-glucan synthase complex was capable of synthesizing both (1----3)- and (1----4)-beta-glucans . A new photoreactive pyrimidine analogue of uridine 5'-diphosphoglucose, 5-{3-(p-azidosalicylamide}allyl-uridine 5'-diphosphoglucose was synthesized in a three-step reaction sequence involving mercuration of UDP-Glc, alkylation of 5-Hg-UDP-Glc, and acylation of 5-(3-amino)allyl-UDP-Glc and characterized by chemical and spectroscopic analysis . The analogue inhibits (Kiapp 16 microM) and, upon UV irradiation, irreversibly inactivates the beta-glucan synthase . The analogue was iodinated with Na125I to give a radiolabeled, photoreactive compound, and was used in photoaffinity labeling of UDP-Glc pyrophosphorylase, UDP-Glc dehydrogenase, and several putative UDP-Glc-binding proteins from L . multiforum . The radiolabeled analogue specifically labeled the 31-kDa polypeptide in the immunoprecipitated synthase complex . The photolabeling of this polypeptide is strictly dependent on UV irradiation, is blocked by uridine 5'-diphosphoglucose and uridine 5'-diphosphate, and reaches saturation at analogue concentrations above 300 microM . These results indicate that the 31-kDa polypeptide in the beta-glucan synthase complex bears a uridine 5'-diphosphoglucose-binding site and is involved in the catalysis of beta-glucan synthesis.

J Immunol Methods, 1991 Nov 22, 144(2), 197 - 202
Detection of staphylococcal enterotoxin and toxic shock syndrome toxin-1 (TSST-1) by immunoblot combined with a semiautomated electrophoresis system; Orden JA et al.; An immunoblot technique combined with a semi-automated electrophoresis system has been developed for the detection of staphylococcal enterotoxins and toxic shock syndrome toxin-1 (TSST-1) . The advantages of this method over other detection techniques include speed, sensitivity (10 ng/ml) and specificity . The use of semiautomated electrophoresis permits the routine detection of staphylococcal enterotoxins and TSST-1.

Proc Natl Acad Sci U S A, 1991 Nov 15, 88(22), 9954 - 8
Identification of the staphylococcal enterotoxin A superantigen binding site in the beta 1 domain of the human histocompatibility antigen HLA-DR; Herman A et al.; The staphylococcal enterotoxin A (SEA) is a superantigen that must bind to class II molecules of the major histocompatibility complex to be recognized by T cells . In humans, most HLA-DR class II allelic and isotypic forms, such as DR1, bind SEA well . DRw53 is an exception, binding SEA very poorly . We have localized this difference to a single residue (amino acid 81) in the beta 1 domain . A highly conserved histidine at residue 81 allows SEA binding, but a tyrosine does not . Residue 81 is predicted to lie in an alpha-helix on the surface of the molecule, with its side chain pointing up out of the pocket associated with binding of conventional peptide antigens . This finding supports the hypothesis that superantigens and conventional antigens bind to different sites on the class II molecule.

FEMS Microbiol Lett, 1991 Nov 15, 68(2), 143 - 9
Suppression of an Escherichia coli secA(ts) mutant by a gene cloned from Staphylococcus carnosus; Overhoff-Freundlieb B et al.; Escherichia coli mutant MM52 (secA(ts)) was transformed with a cosmid library from Staphylococcus carnosus, and a recombinant cosmid (pBO23) allowing growth at the non-permissive temperature (42 degrees C) was isolated . pBO23 also restored the growth defects of E . coli mutants IQ85 (secY(ts)) and IT41 (lep(ts)) . Nucleotide sequencing revealed that the DNA fragment responsible for the suppression effect codes for a S . carnosus protein highly homologous to the ribosomal protein L13 of E . coli . The staphylococcal L13 protein was efficiently incorporated into E . coli ribosomes . Possible explanations for the effect of this polypeptide on the growth of temperature-sensitive E . coli secretion mutants are discussed.

Proc Natl Acad Sci U S A, 1991 Nov 15, 88(22), 10267 - 71
Profound alteration in an alpha beta T-cell antigen receptor repertoire due to polymorphism in the first complementarity-determining region of the beta chain; Gahm SJ et al.; Amino acid residues that are critical in maintaining the framework structure of immunoglobulin heavy- and light-chain variable (V) regions are strongly conserved in the V alpha and V beta proteins of the alpha beta T-cell antigen receptor (TCR alpha beta) . Consequently, it has been proposed that TCR alpha beta has a conformation similar to that of an immunoglobulin Fab fragment and that the regions of the TCR homologous to the three immunoglobulin complementarity-determining regions (CDRs 1, 2, and 3) bind to the peptide antigen-major histocompatibility complex (MHC) molecule ligand . A single amino acid substitution in the predicted CDR1 of the V beta 3 protein of certain mouse strains dramatically altered TCR alpha beta usage in an antigen-specific MHC-restricted immune response but did not abrogate V beta 3 specificity for the superantigens minor lymphocyte stimulatory locus (Mls)c and staphylococcal enterotoxin A (SEA) . The results confirm the importance of the V beta CDR1 in antigen-MHC molecule recognition, supporting the Fab-like structural model of TCR alpha beta, and provide further evidence that conventional antigen-MHC recognition and superantigen recognition are mediated by distinct regions of the TCR beta chain . They also suggest that allelic polymorphism may be a significant source of diversity in the TCR repertoire.

J Immunol, 1991 Nov 15, 147(10), 3274 - 81
Mutation of the disulfide loop in staphylococcal enterotoxin A . Consequences for T cell recognition; Grossman D et al.; The hallmark of T cell responses to staphylococcal enterotoxins (SE) and other super-Ag is a selective stimulation of cells expressing particular TCR-V beta segments . Our previous studies suggested that the disulfide loop in SE is critical for their interaction with the TCR . To investigate this concept in further detail we constructed disulfide loop mutants of staphylococcal enterotoxin A (SEA), and examined these altered toxins for mitogenicity, class II MHC binding, and V beta specificity . We found that substitutions of either Cys-96 or Cys-106 decreased mitogenicity by 100-fold without significantly affecting class II binding or resistance of the molecule to proteolysis . Several mutants lost the capacity to stimulate V beta 11+ cells, except a Cys-106----Gln mutant for which V beta 11-stimulatory activity was increased . By contrast, mutants containing Cys----Ala substitutions acquired the capacity to stimulate V beta 6+ cells . Despite these effects of V beta specificity, all mutants retained the predominant preference of SEA for V beta 3+ cells . Neither exchange of regions flanking the loop in SEA with corresponding residues in SEB, nor conversion of the entire loop region of SEA to that of SEE, were associated with transfers of V beta specificity . Our results suggest that the disulfide loop in SEA contributes to toxin avidity for the TCR, rather than specificity for particular V beta.

J Immunol, 1991 Nov 15, 147(10), 3296 - 302
Pertussis toxin prevents the induction of peripheral T cell anergy and enhances the T cell response to an encephalitogenic peptide of myelin basic protein; Kamradt T et al.; In a murine model of T cell-mediated autoimmune disease, experimental autoimmune encephalitis (EAE), 80% of all encephalitogenic T cell clones in H-2u mice use the V beta 8.2 TCR element . To induce EAE in susceptible strains of mice either heat-killed Bordetella pertussis organisms or Bordetella pertussis toxin (PT) must be injected in addition to Ag in CFA . We investigated the mechanisms by which PT facilitates the induction of EAE . Our data show, that PT interferes with the induction of Ag-induced peripheral T cell anergy . Furthermore it has a specific adjuvanticity for the autoantigen pAc1-11 in vivo and acts as a selective mitogen in vitro . We also tested the hypothesis that PT is a bacterial superantigen that specifically expands the V beta 8.2+ subset of T cells, thereby expanding the encephalitogenic T cell clones that are contained in this subset, so that the number of autoreactive T cells is brought over a critical threshold, necessary to induce autoimmune disease . Our data show that PT is not a superantigen . Staphylococcal enterotoxin B, a V beta 8.2-specific superantigen, does not enhance the immune response to the encephalitogenic peptide.

Am J Respir Cell Mol Biol, 1991 Nov, 5(5), 411 - 5
Human alveolar macrophages inhibit receptor-mediated increases in intracellular calcium concentration in lymphocytes; Yarbrough WC Jr et al.; Prior studies have demonstrated that human alveolar macrophages (AM) are suppressive of lymphocyte function, through the mechanism of inhibition is unclear . In the current study, human AM inhibited receptor-mediated increases in intracellular calcium concentration ({Ca2+}i) in T cells, natural killer cells, and B cells . This effect was produced by either live or fixed AM, while peripheral blood monocytes caused a minimal reduction in {Ca2+}i . The inhibitory effect of AM was seen following 1 to 2 h of incubation with lymphocytes, was complete at 16 h, and did not affect ionomycin-mediated {Ca2+}i . Inhibition of {Ca2+}i by AM correlated with suppression of T-lymphocyte proliferation and cytotoxic T-lymphocyte activity in response to alloantigen and Staphylococcus A-induced immunoglobulin production . Our findings suggest that a membrane signal on AM is capable of inhibiting receptor-mediated signal transduction in lymphocytes and that this is likely a major mechanism by which immune responses are downregulated in the alveolus.

Anesthesiology, 1991 Nov, 75(5), 893 - 5
Drug contamination from opening glass ampules; Zacher AN et al.; Recently there have been several reports of postoperative sepsis due to the intravenous injection of contaminated solutions of propofol (Diprivan) . The mechanism by which this contamination occurred has not been identified . This study sought to determine whether bacterial contamination of the contents of glass ampules can be decreased by swabbing the neck of the vial with alcohol prior to opening . Glass ampules of 1% propofol and 1% lidocaine were swabbed with a solution of Staphylococcus epidermidis . Half of these ampules were subsequently wiped with alcohol pads prior to being opened . An aliquot from each ampule was pipetted into a nutrient broth and allowed to incubate overnight at 37 degrees C . These solutions were plated on agar, incubated for 24 h, and then examined for bacterial growth . Three of the eight lidocaine ampules and six of the eight propofol ampules not cleaned with alcohol demonstrated evidence of bacterial contamination . The contents of all ampules that had been wiped with alcohol prior to being opened remained sterile (P less than 0.001 vs . non-alcohol-treated group for propofol ampules and P = 0.20 vs . non-alcohol-treated group for lidocaine ampules) . These data suggest that bacterial contamination of propofol and lidocaine may occur as a result of opening glass ampules . Wiping the outside of the ampule with alcohol immediately prior to opening may decrease this risk.

South Med J, 1991 Nov, 84(11), 1375 - 7
Echocardiographic diagnosis of effusive-constrictive pericarditis due to staphylococcal pericarditis after cardiac surgery; D'Cruz IA et al.; Two weeks after coronary artery bypass surgery, a 43-year-old man was readmitted with fever, pneumonia, left pleural effusion, and pericarditis . Echocardiography showed a localized posterior pericardial effusion, pericardial thickening, and bulging of the ventricular septum toward the left ventricle . Right-sided catheterization indicated pericardial constriction . Effusive-constrictive pericarditis was confirmed at surgery . Cardiac imaging played an important role in diagnosis of this unusual complication of cardiac surgery.

Infect Immun, 1991 Nov, 59(11), 4049 - 55
Murine macrophage activation by staphylococcal exotoxins; Fleming SD et al.; We investigated the ability of staphylococcal enterotoxins A and B, exfoliative toxins A and B, and toxic shock syndrome toxin 1 to activate macrophages . All of the toxins tested had the potential to stimulate tumoricidal activity in peritoneal macrophages from lipopolysaccharide-responsive C3HeB/FeJ mice . In contrast, none of the toxins activated cytotoxicity in lipopolysaccharide-unresponsive macrophages from C3H/HeJ mice . We also studied toxin stimulation of monokine secretion . Staphylococcal enterotoxin A, toxic shock syndrome toxin 1, and both exfoliative toxins triggered C3HeB/FeJ macrophages to secrete tumor necrosis factor alpha, but enterotoxin B induced only marginal amounts of tumor necrosis factor . All of the toxins used stimulated interleukin-6 production by macrophages from both strains of mice . Nitric oxide is produced in response to the exfoliative toxins only by the lipopolysaccharide-responsive macrophages . These results suggest that macrophages respond differently to several staphylococcal exotoxins.

Eur J Immunol, 1991 Nov, 21(11), 2703 - 9
Interferon-gamma and growth factor production by murine T cells derived from three different lymphoid tissues; Hoiden I et al.; Various antigen-presenting cells and the environment in different lymphoid tissues have been suggested to influence the type of lymphokine produced by T cells . We have investigated the mitogen-induced proliferation, interferon-gamma (IFN-gamma) and growth factor production by cells isolated from spleen, mesenteric and peripheral (axillary, brachial and inguinal) lymph nodes (LN) . We found that stimulation with concanavalin A or staphylococcus enterotoxin B induced IFN-gamma synthesis in spleen cells but not in LN cells . Proliferation and growth factor production were comparable in the three cell populations . The addition of the phorbol ester phorbol 12-myristate 13-acetate (PMA), which is commonly used as a substitute for accessory cells, did not influence the IFN-gamma synthesis by LN cells . The growth factor production was, on the other hand, elevated by the addition of PMA . A high number of IFN-gamma-producing peripheral LN cells were obtained if they were stimulated in the presence of splenic adherent cells . The growth factor synthesis was marginally affected by the presence of these cells . Thus, splenic adherent cells provide a co-stimulatory signal to the T cell necessary for IFN-gamma synthesis.

Plant Mol Biol, 1991 Nov, 17(5), 1013 - 21
Expression of antifreeze proteins in transgenic plants; Hightower R et al.; The quality of frozen fruits and vegetables can be compromised by the damaging effects of ice crystal growth within the frozen tissue . Antifreeze proteins in the blood of some polar fishes have been shown to inhibit ice recrystallization at low concentrations . In order to determine whether expression of genes of this type confers improved freezing properties to plant tissue, we have produced transgenic tobacco and tomato plants which express genes encoding antifreeze proteins . The afa3 antifreeze gene was expressed at high steady-state mRNA levels in leaves from transformed plants, but we did not detect inhibition of ice recrystallization in tissue extracts . However, both mRNA and fusion proteins were detectable in transgenic tomato tissue containing a chimeric gene encoding a fusion protein truncated staphylococcal protein A and antifreeze protein . Furthermore, ice recrystallization inhibition was detected in this transgenic tissue.

J Biomol NMR, 1991 Nov, 1(4), 439 - 46
Measurement of 15N-13C J couplings in staphylococcal nuclease; Delaglio F et al.; 15N-C alpha and 15N-C' J couplings were measured for the backbone of staphylococcal nuclease, uniformly enriched with 15N and 13C . It is found that the 1JC'N coupling is similar for beta-sheet, J = 14.8 +/- 0.5 and for alpha-helix, J = 14.8 +/- 0.4 but tends to be larger for the unstructured N- and C-terminal ends of the protein (J = 15.6 +/- 0.5) . On average, 1JNC alpha are smaller for alpha-helical residues (J = 9.6 +/- 0.3 Hz) compared to beta-sheet (J = 10.9 +/- 0.8 Hz) and a substantial difference is observed for 2JNC alpha in alpha-helices (J = 6.4 +/- 0.4 Hz) and beta-sheets (J = 8.3 +/- 0.8 Hz).

J Immunol, 1991 Nov 1, 147(9), 3080 - 6
Differential sensitivity of virgin and memory T lymphocytes to calcium ionophores suggests a buoyant density separation method and a model for memory cell hyporesponsiveness to Con A; Miller RA et al.; Previous work from this laboratory has indicated that murine memory T cells differ from virgin T cells in that the former are more resistant to agents that alter intracellular {Ca}i . We have used this difference to devise a method for separating virgin from memory T cells by centrifugation over an ionomycin-containing Percoll step gradient after brief exposure to 2 microM ionomycin . Under these conditions, those T cells that are most sensitive to ionomycin-induced changes in {Ca}i become more dense and therefore travel further into the Percoll/ionomycin gradient than cells that are more resistant to ionomycin . We show that the ionomycin-resistant cell population is enriched for cells that express high levels of Pgp-1 (CD44), and low levels of CD45RB, and thus appears to consist largely of memory T cells . Both CD4 and CD8 cells can be divided into Pgp-1hi and Pgp-1lo subsets in this way . Cells recovered from such a gradient and washed to remove the ionomycin appear normally functional, i.e., neither more nor less responsive to mitogens and costimuli than untreated cells . Limiting dilution methods show that the ionomycin-sensitive (virgin) subset contains most of the Con A-responsive precursors for cytotoxicity, and most of the cells able to produce IL-2 in responses to Con A or staphylococcal enterotoxin B . Ag-specific helper memory cells are, however, found predominantly in the ionomycin-resistant fraction of the spleen and draining lymph nodes of mice infected with Schistosoma mansoni . Changes in resistance to calcium signal development may represent a fundamental distinction between virgin and memory T cells, and could contribute to differences in activation requirements between these two cell subsets.

Enferm Infecc Microbiol Clin, 1991 Nov, 9(9), 543 - 6
{Effect of subinhibitory concentrations of beta-lactams on the production of slime, surface hydrophobicity and adhesion of Staphylococcus epidermidis}; Martinez-Martinez L et al.; The activity of subinhibitory concentrations of oxacillin, imipenem, cefalotin, cefuroxime, cefotaxime and amoxycillin-clavulanic acid upon slime production, surface hydrophobicity and polystyrene adherence of S . epidermidis was evaluated . Slime production and polystyrene adherence of three Staphylococcus epidermidis strains was assessed using test tube (safranin stained film) or in microtiter plate . The surface hydrophobicity was determined using a biphasic system (water/p-xylene) . Using one-half and one-quarter of MIC for all antibiotics tested, a reduction in slime production and polystyrene adherence was observed in all three strains used . Regarding surface hydrophobicity, in one of the strains used we have observed a rise in surface hydrophobicity values as compared to control strains whenever one-half of MCI was used for all six antimicrobial agents and also with one-quarter of amoxicillin-clavulanate MCI . Subinhibitory MCI of all six antibiotics tested inhibits slime production and adherence of Staphylococcus epidermidis . This inhibitory effect is not related to changes in surface hydrophobicity.

Zh Mikrobiol Epidemiol Immunobiol, 1991 Nov, (11), 45 - 7
{The choice of a method of isolating human IgG F(ab)2 fragments for determining antibodies to staphylococcal protein A}; Minster AB et al.; The comparative study of two methods of the proteolysis of IgG with the aim of obtaining F(Ab)2-fragments from the blood sera of patients and healthy donors has been made . Different methods for the isolation of F (ab)2-fragments, such as gel filtration on Sephadex G-200, affinity chromatography on protein A-BrCN-sepharose, reprecipitation with zinc sulfate, have been analyzed . For further work the method of the peptic decomposition of whole serum with subsequent salting out with ammonium sulfate and gel filtration on Sephadex G-200 has been chosen.

Immunobiology, 1991 Nov, 183(5), 433 - 50
Crosslinked staphylococcal enterotoxin B stimulates CD8+ T cells only in the presence of unlinked costimulator signals; Miethke T et al.; The superantigen staphylococcal enterotoxin B (SEB) binds to class II MHC expressing cells and subsequently causes selective activation of T cells carrying appropriate T cell receptor (TCR) V beta chains . Apparently SEB acts as a bifunctional molecule by bridging class II MHC structures with the appropriate TCR-V beta chains . This assumption predicts that immobilized SEB ought to stimulate purified, class II MHC negative murine T cells . We show here that immobilized SEB lacks the ability to trigger murine CD8 T cells . Responsiveness obtained at a high T cell concentration is due to contaminating class II MHC-positive lymphocytes . Complementation of the culture system with syngeneic irradiated B cells blasts effectively restores responsiveness . The proliferating cells exhibit SEB specific cytotoxicity and a bias for V beta 8 expression . Since no evidence for leakiness of SEB covalently bound to sephadex beads was obtained, the data imply that immobilized SEB in fact binds to the TCR of T cells expressing the appropriate V beta chains . However, for primary activation additional costimulatory signals are required which can be provided in an unlinked fashion by activated B cells . Resting B cells are activated by immobilized SEB to cells expressing high costimulator activity . As such, the data point out a third function of SEB.

J Antimicrob Chemother, 1991 Nov, 28(5), 681 - 94
Screening tests for the detection of methicillin resistance in Staphylococcus epidermidis; Hedin G et al.; Methods to detect resistance to methicillin in Staphylococcus epidermidis were studied in order to find a rapid screening test suitable for routine use . One hundred and forty-nine clinical isolates, 16 isolates from skin of healthy people and two reference strains were studied . Hypersecretion of beta-lactamase as a cause of methicillin resistance was eliminated in the strains studied . Tube and microtitre breakpoint, agar breakpoint and disc diffusion methods were compared . The breakpoint for methicillin resistance used was 16 mg/L in broth and 10 mg/L in agar . The discs used contained 1 and 5 micrograms oxacillin and 5 and 10 micrograms methicillin . Turbidimetric measurements in broth during incubation were carried out using the Bioscreen analysing system . The skin strains were founf to be susceptible in all tests . Using an inoculum of 10(7) cfu/mL 111/149 clinical isolates were classified as resistant after incubation for 24 h at 35 degrees C using the tube and microtitre breakpoint tests, incubation for 72 h did not increase this rate . When an inoculum of 10(5) cfu/mL was used 73% of these strains were identified within 24 h and all within 72 h with the tube breakpoint test . Using the microtitre breakpoint test, with an inoculum of 10(5) cfu/mL or lower, all resistant strains were not detected within 48 h . All agar breakpoint tests required 48 h incubation for reliable results . Only the 1 microgram oxacillin disc always separated strains found to be resistant or susceptible in the tube breakpoint test . The zone of inhibition was clearly readable after 16 h of incubation at 35 degrees C.

Can J Microbiol, 1991 Nov, 37(11), 877 - 80
Assessment of two commercial agglutination kits for detecting Escherichia coli heat-labile enterotoxin; Speirs J et al.; Two commercial agglutination kits, a reserved passive agglutination test (VET-RPLA) and a staphylococcal coagglutination test (Phadebact ETEC-LT Test), were compared with two cell culture assays (Y-1 and Vero) and GM1 ganglioside enzyme-linked immunosorbent assay (GM1-ELISA) for sensitivity in detecting Escherichia coli heat-labile enterotoxin (LT) . Of 48 toxigenic strains, 23 were positive by all assays . One strain was negative only by the Phadebact test . Four strains, all LT-II producers, were positive by cell culture only . For LT-I detection, the Phadebact test was the least sensitive but was simple and rapid; VET-RPLA was simple, sensitive, and a good substitute for cell culture or GM1-ELISA.

FEMS Microbiol Lett, 1991 Nov 1, 68(1), 23 - 6
Characteristics of extracellular protein production by a plasmidless derivative of Staphylococcus simulans biovar staphylolyticus; Nitterauer JD et al.; A derivative of Staphylococcus simulans biovar staphylolyticus cured of all five plasmids present in the wild-type organism was developed, and the characteristics of extracellular protein production by this plasmidless strain were compared to those of the wild type . Although staphylolytic endopeptidase (lysostaphin) and beta-lactamase are known to be plasmid encoded, analysis of this cured strain revealed that most other extracellular proteins are chromosomally encoded.

FEMS Microbiol Lett, 1991 Nov 1, 68(1), 119 - 22
Characterization of staphylococcal plasmids hybridizing with the fosfomycin resistance gene fosB; Etienne J et al.; The distribution of the fosB gene, coding for fosfomycin resistance, in 105 fosfomycin-resistant isolates of Staphylococcus from various geographical areas, was studied by Southern blot hybridization . Nucleotide sequences related to fosB were detected in 36 strains belonging to five species . Plasmids bearing fosB were often of a size similar to that of pIP1842 (2.54 kb) in S . epidermidis, most often small (2.4 to 4.1 kb) in other species including S . aureus where a 2.7-kb plasmid was found in 16 out of the 18 strains studied . The fosB gene was geographically dispersed since it was present in six different locations in France and also in Japan . The weak hybridization observed with plasmid DNA of certain strains of S . aureus, S . epidermidis, S . haemolyticus, S . saprophyticus, and S . warneri may indicate gene heterogeneity for fosfomycin resistance in Staphylococcus spp.

Nippon Seikeigeka Gakkai Zasshi, 1991 Nov, 65(11), 1112 - 9
{Haematogenous osteomyelitis induced by strain SMU-0611 of Staphylococcus warneri in mice}; Matsuoka M; Strain SMU-0611 of Staphylococcus warneri isolated from clinical specimens (1.25 x 10(9) colony forming units) was intravenously administered into mice . Strain SMU-0611 caused remarkable inflammation characterized by roentgenographically recognizable changes in bone at 14 days after the injection . A number of organisms were detected in the bone marrow for as long as 35 days after the injection, suggesting specific lodgement of the strain in mouse bone marrow . Concerning the X-ray features of the mice, multiple foci of osteolytic changes were shown in the metaphysis of the femur and the tibia obtained 14 days after the injection . However, these changes did not aggravate at all for 56 days after the injection.

DNA Cell Biol, 1991 Nov, 10(9), 663 - 9
Restriction fragment fingerprint and genome sizes of Staphylococcus species using pulsed-field gel electrophoresis and infrequent cleaving enzymes; Poddar SK et al.; Large restriction fragments of genomic DNA from Staphylococcus species were separated by pulsed-field gel electrophoresis (PFGE) . Five different strains of S . aureus (ISP8, SAU3A, PS96, ATCC 6538, ATCC 15564) and three representative strains of S . haemolyticus SM102, S . warneri MCS4, S . cohnii LK478 from human hosts, and one strain of S . aureus (ATCC 8432) from an avian host were used in this study . Since Staphylococcus is A + T rich (approximately 67%), restriction fragments were obtained by digesting chromosomal DNA with endonucleases that recognize GC-rich sequences . Five enzymes Csp I, Sma I, Ecl XI, Ksp I, or Sac II were used for generation of few (7 to 16) distinctly separated fragments, with average sizes in the range of 200-300 kb . The size distribution of restriction fragments for each enzyme for each strain produced a strain-identifying fingerprint, and the genome size of each strain was determined from such restriction fragments separated by PFGE.

J Clin Ultrasound, 1991 Nov-Dec, 19(9), 555 - 60
Superficial masses with color flow Doppler imaging; Mitchell DG et al.; Color Doppler imaging findings and spectral waveforms were reviewed for 21 superficial masses in which color Doppler imaging revealed flow . This series included 5 benign hyperplastic lymph nodes, 5 malignant masses, 2 staphylococcus abscesses, and 9 pseudoaneurysms . Benign hyperplastic nodes had a vascular pattern consisting of prominent central flow with a branching radial pattern, reflecting the morphology of lymph nodes . One metastasis from melanoma demonstrated abundant peripheral flow and central avascularity, while the other 4 malignant masses and the 2 abscesses had sparse internal flow . All pseudoaneurysms had a to-and-fro imaging and spectral pattern (into the mass during systole, out during diastole), and/or a swirling pattern on real-time images (8 of 9 each) . Doppler spectral analysis revealed continuous forward flow during diastole for all other masses and abscesses . Waveforms appeared similar for benign and malignant masses, and for abscesses . Conventional image-directed Doppler examinations performed prior to color Doppler studies failed to reveal flow in 2 partially thrombosed pseudoaneurysms, 1 malignant mass, and 1 abscess . Benign hyperplastic nodes, malignant masses, and pseudoaneurysms had distinctive color Doppler image appearances in this series, although demonstration of flow within a mass does not eliminate the possibility of abscess . Proper use of color and spectral Doppler analysis of superficial masses involves more than mere detection or exclusion of flow.

J Clin Invest, 1991 Nov, 88(5), 1531 - 9
Escherichia coli hemolysin is a potent inductor of phosphoinositide hydrolysis and related metabolic responses in human neutrophils; Grimminger F et al.; Escherichia coli hemolysin (Hly) is a proteinaceous pore-forming exotoxin that probably represents a significant virulence factor in E . coli infections . We investigated its influence on human polymorphonuclear neutrophils (PMN), previously identified as highly susceptible targets . Hly provoked rapid secretion of elastase and myeloperoxidase, generation of superoxide, and synthesis of platelet-activating factor (PAF) and lyso-PAF . Concomitantly, marked phosphatidylinositol (PtdIns) hydrolysis with sequential appearance of the inositol-phosphates, inositol-phosphates, inositol triphosphate, diphosphate, and monophosphate, respectively, and formation of diacylglycerol, occurred . The metabolic responses displayed distinct bell-shaped dose dependencies, with maximum events noted at low toxin concentrations of 0.1-0.5 hemolytic units per milliliter . PtdIns hydrolysis and metabolic responses elicited by Hly exceeded those evoked by optimal concentrations of formylmethionyl-leucyl phenylalanine, PAF, leukotriene B4, A23187, or staphylococcal alpha-toxin . The toxin-induced effects were sensitive toward modulators of PMN stimulus transmission pathways (pertussis toxin, the kinase C inhibitor H7, and phorbol myristate acetate "priming") . We conclude that the marked capacity of low doses of Hly to elicit degranulation, respiratory burst, and lipid mediator generation in human PMN probably envolves signal transduction via PtdIns hydrolysis.

Biochem Biophys Res Commun, 1991 Oct 31, 180(2), 742 - 7
Identification of a site for carboxyl methylation in human alpha-globin; Ladino CA et al.; The human erythrocyte protein carboxyl methyltransferase modifies unusual protein D-aspartyl and L-isoaspartyl residues which arise spontaneously from internal rearrangements accompanying asparaginyl deamidation and aspartyl isomerization . A site of methylation associated with alpha-globin in intact cells has been identified by peptide mapping of radiolabeled globin isolated from human erythrocytes previously incubated with L-{methyl-3H}methionine . The site is located in a Staphylococcus V8 peptide containing residues 1-30 of alpha-globin . Two potential sources of methylation sites are present in this sequence at Asp-t and Asn-9.

Proc Natl Acad Sci U S A, 1991 Oct 15, 88(20), 9287 - 91
Monoclonal antibody-targeted superantigens: a different class of anti-tumor agents; Dohlsten M et al.; The bacterial superantigen staphylococcal enterotoxin (SE) A (SEA) directs cytotoxic T lymphocytes (CTLs) expressing particular sequences of the T-cell receptor (TCR) beta chain to lyse tumor cells expressing major histocompatibility complex (MHC) class II molecules, which serve as receptors for SEs . We now report that chemical conjugates of SEA and the colon carcinoma-reactive monoclonal antibodies (mAbs) C215 or C242 mediate T cell-dependent destruction of colon carcinoma cells lacking MHC class II molecules . SEA was covalently linked to the mAbs C215 and C242 via a PEG-based hydrophilic spacer . The C215-SEA conjugate targeted CD4+ as well as CD8+ CTLs to lyse a panel of colon carcinoma cells lacking MHC class II molecules . T-cell recognition of mAb-SEA conjugates was SEA specific, since SEB-selective T-cell lines with potent cytotoxic activity towards Raji cells coated with SEB did not respond to the C215-SEA conjugate . Unconjugated SEA did not induce T-cell lysis of MHC class II- colon carcinoma cells but efficiently directed CTLs against MHC class II+ Raji cells and certain interferon-treated MHC class II+ colon carcinoma cells . These results suggest that SEA-mAb conjugates retain the SEA-related selectivity for certain TCR beta-chain variable region (V beta) sequences but, in contrast to unconjugated SEA, mediate the TCR interaction in a MHC class II-independent manner . The cytotoxic activity mediated by C215-SEA and C242-SEA conjugates was blocked by excess of C215 mAb and C242 mAb, respectively, showing that the specificity in the targeting of mAb-SEA conjugates is defined by the antigen reactivity of the mAb . These results demonstrate that bacterial superantigens may be successfully conjugated to mAb with preserved T cell-activating capacity . The circumvention of MHC class II binding of SEs by conjugation to mAb suggests that such conjugates may find general application as antitumor agents, taking advantage of the extreme T cell-activating potency of superantigens.

Cornell Vet, 1991 Oct, 81(4), 389 - 95
Antibiotic-responsive generalized nonlesional pruritus in a dog; Miller WH Jr; A 15-year-old dog was evaluated for a nonresponsive generalized pruritic condition of 5 months duration . Routine diagnostic testing, including intradermal testing with 63 inhaled allergens and the feeding of a home-cooked hypoallergenic diet, failed to define the cause of the pruritus . An intradermal skin test with a staphylococcal cell wall/toxoid mixture and a skin biopsy of the skin test site suggested that the dog had a bacterial hypersensitivity . Antibiotic therapy eliminated the pruritus and the dog's pruritus was successfully managed for 3 years with the combined use of subtherapeutic dosages of antibiotics and a commercial staphylococcal vaccine.

Nippon Kyobu Geka Gakkai Zasshi, 1991 Oct, 39(10), 1932 - 6
{Mitral prosthetic valve endocarditis diagnosed by transesophageal echocardiography--a surgical case report}; Shimono T et al.; The patient was a 39-year-old female who had undergone mitral valve replacement 2 years previously . Staphylococcus epidermidis was detected on blood culture . Transesophageal echocardiography revealed vegetation attached to the left atrial site of the mitral prosthesis, whereas precordial echocardiography did not reveal any abnormal echo structure . The patient was performed an emergency operation, during which the echocardiographic finding was confirmed . We conclude that performance of transesophageal echocardiography is useful when clinical features suggest prosthetic valve endocarditis.

J Clin Pathol, 1991 Oct, 44(10), 873 - 4
Staphylococcus lugdunensis; Barker KF et al.; A fatal case of penicillin resistant Staphylococcus lugdunensis endocarditis on a native valve damaged by rheumatic fever is described . A retrospective examination of 978 tube coagulase negative staphylococcal blood culture isolates was also performed to determine how often this species is isolated . None of the 978 isolates was identified as S lugdunensis . It is concluded that ornithine decarboxylase production is the only reliable method of identifying this species.

J Appl Bacteriol, 1991 Oct, 71(4), 347 - 53
Conversion of biological and immunological properties during a process of decapsulation in a strain of Staphylococcus hyicus; Yoshida E et al.; Repeated subculture at 42 degrees C of Staphylococcus hyicus strain ST67P, which exhibits streaming-type growth in a soft-agar medium, yielded three variants, ST67L, ST67S and ST67C, which had different colonial morphologies; small compact colonies possessing long and short tails and perfect compact colonies . The parent strain and ST67L respectively gave strong and weak positive intensity when stained by rabbit antisera prepared by capsular type I and II strains of Staph . epidermidis conjugated with fluorescein isothiocyanate . Variant ST67L gave a positive result with antiserum prepared by capsular type I strain and no staining was observed with variants ST67S and ST67C against these antisera preparations . Strain ST67C had the lowest virulence although no remarkable difference was shown between the parent strain and variants ST67L and ST67S . The cell volume index of the parent strain was 1.35, 2.43 and 3.71 times larger than those of ST67L, ST67S and ST67C, respectively . The converting activity of rabbit anti-ST67P strain serum absorbed by strain ST67C required four times more of the organisms than strain ST67P, changing the colonial morphology of the strain from diffuse to compact type by the addition of antiserum to soft agar medium . Positive coagulase and false positive clumping factor reaction were shown in variants ST67C, but no remarkable alteration was observed with 19 biochemical properties determined by a conventional identification kit . In ulta-thin sections of the parent strain labelled with rabbit anti-strain ST67P serum conjugated with ferritin, large capsule surrounded by ferritin granules were demonstrated by electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Br J Clin Pharmacol, 1991 Oct, 32(4), 433 - 9
Nicardipine sustained release in hypertension; Webster J et al.; 1 . A novel formulation of nicardipine (25% standard, 75% sustained release--SR) was evaluated in mild hypertension in a double-blind, randomized, placebo-controlled comparison with standard nicardipine (STD), using clinic measurements (Hawksley) augmented by home recorded blood pressures (Copal UA 251) . 2 . At 2 h after dosing (peak effect) both STD nicardipine (30 mg three times daily) and SR nicardipine (60 mg twice daily) for 28 days produced a highly significant reduction in sitting and standing blood pressure . The mean sitting blood pressure was reduced by 20/16 mm Hg (STD) and by 25/18 mm Hg (SR) compared with placebo . 3 . Predose (8-11 h after last dose of STD, 12-15 h after last dose of SR) the reductions in sitting blood pressure relative to placebo were 11/6 mm Hg (STD) and 14/7 mm Hg (SR) . 4 . Home recordings confirmed the hypotensive effect of both formulations . Both exhibited a distinct 'peak dose' effect between 1-3 h after dosing . The effect of the SR formulation was sustained throughout the 12 h dosing interval . 5 . Of the 60 patients entering the study, one died of unexplained staphylococcal septicaema, two were withdrawn for non drug-related reasons and 14 (32%) were withdrawn because of adverse effects on active therapy (headaches, facial flushing, leg oedema, chest pain, dizziness) . 6 . In the 43 patients who completed the study adverse symptoms were reported more frequently while they were on the two active formulations of nicardipine compared with placebo . Most of these reactions were again of vasodilator origin.(ABSTRACT TRUNCATED AT 250 WORDS)

Ethiop Med J, 1991 Oct, 29(4), 149 - 61
The incidence of methicillin resistant S . aureus strains in clinical specimens in relation to their beta-lactamase producing and multiple-drug resistance properties in Addis Abeba; Geyid A et al.; From December 1987 to July 1988, a total of 17,142 clinical specimens were examined at the National Research Institute of Health (NRIH), Addis Abeba, to determine the incidence of methicillin resistant S . aureus (MRSA) strains . Coagulase positive Staphylococcus spp . were isolated from 355 specimens . Two hundred and forty-nine 249 (70%) isolates were tested for methicillin resistance by minimum inhibition concentration method (MIC) of which 76 (30.5%) were found to be MRSA and 173 (69.5%) were methicillin sensitive S . aureus (MSSA) strains . The presence of beta-lactamase production was determined in the 355 S . aureus isolates using the rapid Iodometric and Nitrocefin methods and 252 (71%) isolates were found to be beta-lactamase procedures . Furthermore, 47 (62%) of the 76 MRSA isolates and 140 (81%) of the 173 MSSA isolates were beta-lactamase positive strains . The sensitivity pattern of all the S . aureus isolates against 11 common drugs indicated that the majority (80%) of the MRSA strains were multipledrug resistant while 4 (8%) were not resistant to any of the drugs tested . Among the antibiotics, vancomysin and clindamycin were effective against all S . aureus isolates . It was also found that 41 (54%) of the MRSA strains were both betalactamase producers and multiple-drug resistant isolates; of the latter, 36 (87.8%), strains were isolated from pus specimens of patients with post-operative wound infections . The results of this study show that MRSA strains are quite prevalent among specimens referred to the NRIH.

Int J Cardiol, 1991 Oct, 33(1), 83 - 8
Profile of right-sided endocarditis: an Indian experience; Grover A et al.; The clinical profile of right-sided infective endocarditis in India was studied from a review of records of patients with infective endocarditis admitted to this hospital . From November 1982 to November 1989, 109 patients with infective endocarditis showed vegetations on cross-sectional echocardiography confirming the diagnosis of infective endocarditis . In 19 (17.4%) patients, only the right side of the heart was involved: specifically the tricuspid valve alone in 10; tricuspid and pulmonary valves in 4; tricuspid valve and right ventricular outflow tract in 1; tricuspid valve and right ventricular free wall in 1; pulmonary valve alone in 2; and bifurcation of pulmonary trunk in 1 . Eleven patients (57.9%) had underlying congenital heart disease whereas the remaining 8 patients (42.1%) did not have any underlying heart disease . The latter group, therefore, had isolated right-sided infective endocarditis . Previous illnesses leading to isolated right-sided infective endocarditis were: puerperal sepsis in 4; septic abortion in 1; staphylococcal pneumonia in 2; and epididymoorchitis in one . Eight out of 11 patients with congenital heart disease did not report any previous illness . In the remaining 3, right-sided endocarditis followed cardiac surgery in one; dental extraction without prophylaxis in one; and pulmonary balloon valvoplasty in one . All patients with isolated right-sided infective endocarditis had features of septicaemia, but a murmur of tricuspid regurgitation was audible in only 4 (50%) of them . We conclude that, unlike western reports, the pattern of right-sided infective endocarditis in India is different . No drug addict with right-sided infective endocarditis was seen; puerperal sepsis and septic abortion were the commonest causes of isolated right-sided infective endocarditis.(ABSTRACT TRUNCATED AT 250 WORDS)

Ann Hematol, 1991 Oct, 63(4), 223 - 6
Immunological characterization of canine hematopoietic progenitor cells; Hahn J et al.; Canine hematopoietic progenitor cells were characterized by separation with monoclonal antibodies . Depleted and enriched fractions were studied for growth of CFU-GM in semisolid agar and for repopulating capacity of lethally irradiated dogs . CFU growth was not reduced by depletion of marrow using monoclonal antibodies F 3-20-7 (anti-dog Thy-1), MT606 (anti-human CD 6), and IOT2a (anti-human DR) . CFU growth was variable following treatment with the anti-canine T-cell antibody MdT-P 1 and immunomagnetic bead separation . It was regularly enriched when MdT-P 1 treatment was followed by immunorosetting with staphylococcal protein A-loaded sheep red blood cells and density gradient separation . Lethally irradiated dogs were reconstituted by autologous marrow depleted of MdT-P 1-positive cells using immunorosetting and density gradient centrifugation, whereas immunomagnetic bead-depleted marrow was ineffective . Fluorescence-activated cell sorting showed enrichment of hematopoietic progenitor cells in the weakly MdT-P 1-positive fraction.

Postgrad Med, 1991 Oct, 90(5), 58 - 66, 69-73
Pneumonia . Pearls for interpreting patients' radiographs; Montgomery JL; The chest radiograph can detect pneumonia, but laboratory evaluation is needed to determine the specific causative organism . However, before these results become available, a presumptive diagnosis can be made with the help of chest radiography . A lobar pattern on radiography is usually produced by pneumococcal and Klebsiella infections . A lobular (bronchopneumonia) pattern may be produced by Staphylococcus, gram-negative organisms, and anaerobes . An interstitial pattern results from viral, Mycoplasma, and Pneumocystis carinii infections . Mixed patterns may also occur . There is great variation in the presentation of each infection . However, pertinent clinical information, epidemiologic factors, and associated radiographic findings (eg, adenopathy, pleural effusion, cavitation) are helpful in further limiting diagnostic possibilities.

Infect Immun, 1991 Oct, 59(10), 3667 - 72
Influence of major histocompatibility complex haplotype on the mitogenic response of T cells to staphylococcal enterotoxin B; Robinson JH et al.; The abilities of antigen-presenting cells (APC) from nine independent major histocompatibility complex haplotypes and a number of intra-H-2 recombinant congenic strains of mice to present staphylococcal enterotoxin B (SEB) and induce proliferation in murine T-cell receptor V beta 8+ T-cell clones were compared . SEB presented by APC of all haplotypes tested induced significant responses in each of the T-cell clones . The magnitude of response was similar for most haplotypes, but there were limited quantitative differences between certain haplotypes . SEB presented by APC from H-2b mice as well as the intra-H-2 recombinant strains B10.GD and B10.A(4R), which do not express cell surface I-E (designated I-E-), induced the poorest T-cell responses . However, APC from AfE-, AsE-, and AqE- mice were as potent in SEB presentation as APC expressing both I-A and I-E . Antibodies against I-E were more effective than anti-I-A antibodies at inhibiting responses to SEB presented by APC expressing both I-A and I-E, whereas responses induced by APC expressing I-A but not I-E were blocked by antibodies against I-A . Thus, our results show that I-A can present SEB efficiently but that expression of both I-A and I-E on the same APC results in presentation of SEB predominantly by I-E . In addition, experiments using four distinct I-E- strains of mice indicate that I-A alleles differ in their ability to present SEB.

Cell Immunol, 1991 Oct 1, 137(1), 61 - 71
Induction of interleukin-1 in human monocytes by the superantigen staphylococcal enterotoxin A requires the participation of T cells; Gjorloff A et al.; Nanogram quantities of the bacterial superantigen Staphylococcal Enterotoxin A (SEA) induced significant amounts of extracellular IL-1 alpha and IL-1 beta in human peripheral blood mononuclear cells . Induction of maximal IL-1 alpha and IL-1 beta levels by lipopolysaccharide (LPS) required microgram quantities . LPS induced detectable extracellular IL-1 content within 3-6 hr and maximal levels were detected already after 12 hr . Induction of IL-1 production by SEA showed a delayed release with peak values after 24-48 hr . IL-1 beta was the major species of IL-1 seen in both SEA- and LPS-stimulated culture supernatants . SEA was in general a relatively stronger inducer of extracellular IL-1 alpha than LPS . SEA-induced extracellular IL-1 production in human monocytes was entirely dependent on the presence of T cells, whereas addition of T cells to LPS-stimulated purified human monocytes only marginally enhanced the extracellular IL-1 production . The capacity to induce extracellular IL-1 production in monocytes in response to SEA was high in the CD4+ 45RO+ memory T cell subset, whereas CD4+ 45RA+ naive T cells and CD8+ T cells had lower IL-1-inducing capacity . The T cell help for IL-1 production could not be replaced by a panel of T cell-derived recombinant lymphokines added to SEA-stimulated monocytes, including IFN-gamma and TNF, indicating the participation of cell membrane-bound ligands or hitherto unidentified soluble mediators.

J Chemother, 1991 Oct, 3(5), 315 - 20
An open study of teicoplanin in the treatment of gram-positive infections; Nathwani D et al.; Teicoplanin, a new glycopeptide antibiotic similar to vancomycin, was evaluated in treating 36 hospitalized patients suffering from various Gram-positive infections . The 36 patients received teicoplanin once daily as a mean intravenous injection of 550 mg/day (range 200-800 mg/day) . Previous antimicrobial therapy was used in 28% of patients . The mean duration of therapy was 7.5 days (range 3-38 days) . The overall clinical success rate was 94% . 24/36 patients (66%) had positive microbiology . Elimination of the pathogens was seen in 75% of all evaluable cases . Four patients with early prosthetic valve endocarditis due to coagulase negative Staphylococcus (3 patients) and Propionibacterium acnes (1 patient) had a favorable clinical and microbiological outcome . No adverse drug reactions were observed . Teicoplanin is safe and effective in the therapy of many different infections caused by Gram-positive bacteria.

J Am Acad Dermatol, 1991 Oct, 25(4), 674 - 81
Clinical and bacteriologic evaluation of OPC-7251 in patients with acne: a double-blind group comparison study versus cream base; Kurokawa I et al.; Twenty-eight patients with acne were assigned to 4 weeks of treatment with OPC-7251 (a new fluoroquinolone derivative) 1% cream or the cream base in a double-blind manner to evaluate the antibacterial effect of the drug on resident bacteria in the hair follicles and to evaluate clinical response . Propionibacterium acnes was isolated from 21 of the 28 acne patients . When the number of P . acnes was compared before and after treatment, the posttreatment P . acnes count in the OPC-7251 1% cream group was significantly (p = 0.000) reduced compared with that in the cream base group . OPC-7251 1% cream was also significantly (p = 0.019) superior to the cream base in terms of clinical response . P . acnes and Staphylococcus epidermidis isolated from the acne lesions were selected for their susceptibility to various antibacterial agents . The minimal inhibitory concentration of OPC-7251 against P . acnes and S . epidermidis was 0.10 to 0.20 and 0.024 to 0.10 micrograms/ml, respectively, which indicates that the drug has a potent antibacterial effect.

Immunology, 1991 Oct, 74(2), 183 - 90
Role of CD4 molecule in intrathymic T-cell development; Takeuchi Y et al.; We have investigated the role of CD4 molecules in intrathymic T-cell repertoire selection . The administration of monoclonal antibody (mAb) to CD4 in organ culture of murine foetal thymus (FTOC) completely inhibited the development of CD4+8- cells, and additional treatment with anti-class II MHC (Ia) mAb caused no further effects on this inhibition . On the other hand, when the potentially autoreactive cells in Mls-1a mice were monitored by expression of the Mls-1a-reactive V beta 6 gene product of T-cell receptor alpha beta (TcR alpha beta), the treatment with anti-CD4 resulted in the appearance of V beta 6-bearing cells to some extent, but this effect was considerably reinforced by the combinatory use of anti-Ia mAb with anti-CD4 . In a model system where the bacterial superantigen staphylococcal enterotoxin B serves as self-antigen to deplete V beta 8-bearing cells in FTOC, the depletion of V beta 8+ cells was restored partially by anti-CD4 alone but completely by the combination with anti-Ia . These results suggest that CD4 is indispensable for positive selection of all CD4+8- thymocytes, whereas participation of CD4 in negative selection is only partial . It was also observed that the development of TcR alpha beta-bearing cells in the CD4-8- population was inhibited by the treatment with anti-CD4 mAb . In Mls-1a mice, V beta 6-bearing cells were developed in CD4-8+, CD4+8+, and also in CD4-8- populations after anti-CD4 mAb treatment . It is suggested that TcR alpha beta-bearing CD4-8- cells are possibly originated from CD4+ cells and undergo CD4-mediated thymic selection.

J Immunol, 1991 Oct 1, 147(7), 2088 - 93
Differential T cell hyporesponsiveness induced by in vivo administration of intact or F(ab')2 fragments of anti-CD3 monoclonal antibody . F(ab')2 fragments induce a selective T helper dysfunction; Hirsch R et al.; Induction of peripheral T cell anergy associated with stimulation through the TCR complex in vivo has been described in mice using chemically modified APC, staphylococcal enterotoxin B, and intact anti-CD3 mAb . In the latter two models, T cell proliferation, IL-2R expression, and lymphokine production have been demonstrated before subsequent induction of hyporesponsiveness, whereas in the former model, these events have not been observed . To further investigate the relationship between mitogenicity and induction of peripheral hyporesponsiveness, mice were treated with either mitogenic intact anti-CD3 mAb or nonmitogenic F(ab')2 fragments of anti-CD3 mAb . T cells from F(ab')2-treated mice demonstrated a selective decrease in helper functions, with minimal effect on CTL function . Specifically, a marked reduction in ability of Th cells to secrete IL-2 when challenged in vitro with mitogen or alloantigen was observed, which persisted for at least 2 mo after mAb administration and which was independent of T cell depletion . Proliferative function was decreased in CD4+ T cells and could not be fully restored with addition of exogenous IL-2 . A helper defect was also evident in vivo, in that F(ab')2-treated mice were deficient in their ability to reject MHC-disparate skin grafts, and in vivo administration of IL-2 reconstituted their ability to reject skin grafts normally . In contrast, T cells from mice treated with intact mAb demonstrated a significant decrease in both CTL and helper functions . A long term reduction in TCR expression on CD4+ cells from F(ab')2-treated mice, and on both CD4+ and CD8+ cells from intact mAb-treated mice was observed . These findings demonstrate that peripheral T cell hyporesponsiveness can be induced in vivo by binding an identical epitope on the TCR complex in the presence or absence of initial proliferation, lymphokine secretion, or IL-2R expression, and that binding to the same epitope can result in varying long term effects on T cell function.

Indian Pediatr, 1991 Oct, 28(10), 1125 - 8
Gentamicin therapy: medical audit; Bagga A et al.; In order to evaluate the prescription practices on gentamicin, we screened retrospectively the case records of 600 patients admitted to this hospital . Of these, 100 received gentamicin, in combination with other antibiotics, as the initial treatment . Therapy with gentamicin was considered to be indicated in 71 cases; 62% were below 1 year . Patients with neonatal septicemia, meningitis, serious staphylococcal infections and those who were immunocompromised received appropriate treatment with the drug . All the bacteria cultured were sensitive to the antibiotic . Gentamicin was unnecessarily prescribed to 29 cases including bronchial asthma (n = 7), bronchiolitis (n = 6) and acute gastroenteritis with dehydration (n = 8) . In most of the latter cases, the use of gentamicin indicated the psychologic dependence, of the physician, on the drug . Nine patients showed gentamicin-related nephrotoxicity . Formulation of standard treatment guidelines and regular medical audits are necessary to improve quality of prescribing and avoid misuse of drugs.

Biologicals, 1991 Oct, 19(4), 271 - 9
The measurement of staphylococcal protein A by ELISA in immunoglobulin preparations; Lucas C et al.; Chicken antibodies were used to develop an ELISA for the quantitation of parts-per-million levels of protein A in the purification of immunoglobulins or immunoglobulin-like molecules . Quantitation of protein A in the presence of excess human or murine immunoglobulins in this assay was compared with that obtained in ELISAs developed with rabbit antibodies specific either to protein A or to other molecules . Experiments demonstrate that protein A is bound to the immunoglobulins being purified and that this binding affects subsequent recognition by the antibodies used for the assay . Because of these effects and because fragments of protein A might not be detected in assays which rely on Fc binding of protein A, chicken antibodies that bind protein A specifically are an advantage for the quantitation of this protein by ELISA . In addition, comparison of the effect of different types of immunoglobulins on the protein A standard curve suggests that alternatives to including the immunoglobulin under purification with the standards can be utilized.

Rev Clin Esp, 1991 Oct, 189(6), 264 - 7
{Complications of liver transplant in intensive care . Experience in 130 cases}; Cisneros Alonso C et al.; Liver transplant is the first therapeutic choice in most of the advanced liver diseases . Nevertheless, its performance originates a number of complications derived from: a) conservation techniques of the organ (in our study a prolonged time of hot ischemia was significantly associated with); b) surgery (all patients who required massive blood transfusions developed metabolic alkalosis); c) the graft itself (all the F 1 . degrees were significantly infected), and d) extrahepatic causes (cyclosporin was responsible for high blood pressure and nephrotoxicity which appeared as oliguria with good response to furosemide, as well as hyperglycemia) . Some other relevant results in our series were: right pleural effusion and thrombopenia which appeared with a high incidence . Infections were usually originated the staphylococcus which grows in half of the cultures . We also want to highlight the short mean stay and the low mortality incidence in the ICU.

Cryobiology, 1991 Oct, 28(5), 474 - 82
Enhanced survival of yeast expressing an antifreeze gene analogue after freezing; McKown RL et al.; Yeast, like most organisms, survives poorly under freezing conditions . It has been proposed that after rapid cooling yeast suffers a loss in viability from the recrystallization of intracellular ice . Antifreeze proteins found in the blood of certain polar fishes have been shown to be potent inhibitors of ice recrystallization at very low concentrations . We have examined the feasibility of protecting rapidly cooled yeast cells from freezing damage by inhibiting the recrystallization of intracellular ice through in vivo expression of an antifreeze analogue gene . A chemically synthesized gene encoding a protein similar to but differing from the antifreeze proteins of the fish Pseudopleuronectes americanus (winter flounder) was genetically fused to the 3' end of a truncated staphylococcal Protein A gene . When the fused gene was expressed in the budding yeast Saccharomyces cerevisiae, its cells were shown to produce a new chimeric protein that inhibited the recrystallization of ice in vitro . Yeast cells expressing the chimeric antifreeze protein showed a twofold increase in survival after rapid freezing (95 degrees C/min to -196 degrees C) and moderate rates of warming (26 to 64 degrees C/min) compared to cells lacking the chimeric protein.

J Exp Med, 1991 Oct 1, 174(4), 901 - 13
Analysis of T cell stimulation by superantigen plus major histocompatibility complex class II molecules or by CD3 monoclonal antibody: costimulation by purified adhesion ligands VCAM-1, ICAM-1, but not ELAM-1; van Seventer GA et al.; Many ligands of adhesion molecules mediate costimulation of T cell activation . The generality of this emerging concept is best determined by using model systems which exploit physiologically relevant ligands . We developed such an "antigen-specific" model system for stimulation of resting CD4+ human T cells using the following purified ligands: (a) major histocompatibility complex class II plus the superantigen Staphylococcus enterotoxin A, to engage the T cell receptor (TCR); (b) adhesion proteins vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and endothelial leukocyte adhesion molecule 1 (ELAM-1), to provide potential cell surface costimulatory signals; and (c) recombinant interleukin 1 beta (rIL-1 beta)/rIL-6 as costimulatory cytokines . In this biochemically defined system, we find that resting CD4+ T cells require costimulation in order to respond to TCR engagement . This costimulation can be provided by VCAM-1 or ICAM-1; however adhesion alone is not sufficient since ELAM-1 mediates adhesion but not costimulation . The cytokines IL-1 beta and IL-6 by themselves cannot mediate costimulation, but augment the adhesion ligand-mediated costimulation . Direct comparison with the model of TCR/CD3 engagement by CD3 monoclonal antibody demonstrated comparable costimulatory requirements in both systems, thereby authenticating the commonly used CD3 model . The costimulation mediated by the activation-dependent interaction of the VLA-4 and LFA-1 integrins with their respective ligands VCAM-1 and ICAM-1 leads to increased IL-2R alpha (CD25) expression and proliferation in both CD45RA+ CD4+ and CD45RO+ CD4+ T cells . The integrins also regulate the secretion of IL-2, IL-4, and granulocyte/macrophage colony-stimulating factor . In contrast the activation-independent adhesion of CD4+ T cell to ELAM-1 molecules does not lead to T cell stimulation as measured by proliferation, IL-2R alpha expression, or cytokine release . These findings imply that adhesion per se is not sufficient for costimulation, but rather that the costimulation conferred by the VLA-4/VCAM-1 and LFA-1/ICAM-1 interactions reflects specialized accessory functions of these integrin pathways . The new finding that VLA-4/VCAM-1 mediates costimulation adds significance to observations that VCAM-1 is expressed on a unique set of potential antigen-presenting cells in vivo.

J Hosp Infect, 1991 Oct, 19(2), 107 - 14
Investigation of nosocomial prosthetic valve endocarditis due to antibiotic-resistant Staphylococcus epidermidis; Menzies R et al.; A reservoir of antibiotic-resistant Staphylococcus epidermidis strains in our cardiac surgery unit appeared to be the source of organisms responsible for three cases of early prosthetic valve endocarditis . Staphylococcus epidermidis isolates recovered from the skin of 13 patients before and after surgery were compared . All were typed by plasmid profile, antimicrobial susceptibility and slime production . The three strains from early prosthetic valve endocarditis resembled the antibiotic-resistant nosocomial strains recovered from the skin of eight patients following surgery and the environment of the operating theatres . These strains expressed resistance to oxacillin, gentamicin, kanamycin and tobramycin and most produced slime, whereas those isolated from the skin of patients at the time of admission were predominantly susceptible to antibiotics and few produced slime.

Int Immunol, 1991 Oct, 3(10), 1005 - 13
Failure of T cell receptor V beta negative selection in murine intestinal intra-epithelial lymphocytes; Murosaki S et al.; The intestinal intra-epithelial lymphocytes (IEL) are divided into several subsets on the basis of expression of T cell receptor (TCR) alpha beta and gamma delta, intensity of Thy-1 expression and expression of Lyt-3 chain . To investigate the differentiation pathway of the IEL, we examined the repertoire of V beta segments of T cells in the IEL in BALB/c (H-2d, MIs-1b2a) or AKR/J (H-2k, MIs-1a2b) mice . Among freshly isolated IEL, an appreciable number of T cells bearing V beta 3 or V beta 11, which recognize MIs-2a- or MHC IE-encoded molecules respectively, were detected in BALB/c mice . Similarly, in AKR/J mice, IEL contained appreciable levels of V beta 6-bearing T cells . V beta 3- or V beta 11-bearing T cells in the IEL in BALB/c mice increased to a significant level when incubated with staphylococcal enterotoxin A which specifically stimulates V beta 3- and V beta 11-bearing T cells . Most of IEL without clonal deletion expressed Lyt-2 but not Lyt-3 antigens . Such T cells were hardly detected in other organs, including liver . Our results indicate that TCR alpha beta-bearing intestinal IEL that have not undergone negative selection may have differentiated outside the thymus, presumably at a local site of the intestine and can respond normally to the signal via their TCR.

Eur J Immunol, 1991 Oct, 21(10), 2455 - 9
Use of variable human V delta genes to create functional T cell receptor alpha chain transcripts; Sottini A et al.; Previous studies of the human T cell receptor delta genes identified five commonly used V delta segments distinct from any of the known V alpha genes . To define better the relationship between the T cell receptor delta and alpha repertoires we amplified cDNA obtained by polyclonally activated lymphocytes with a common 3' antisense C alpha-specific primer and with five different 5' sense V delta family-specific primers . Amplified products were detected in staphylococcal enterotoxin C2, staphylococcal enterotoxin E, phytohemagglutinin, concanavalin A, anti-CD3 and anti-V beta 8-activated cells, although each cell population expressed a selective pattern of V delta genes . Sequence analysis revealed that each of the known V delta genes can productively rearrange to J alpha segments to produce functional V delta-J alpha-C alpha transcripts . These results argue strongly against the notion that the human V delta and V alpha repertoires are distinct . They further suggest that the restricted delta repertoire observed in many gamma/delta clones results from selection rather than from controlled rearrangements at the T cell receptor alpha/delta locus.

J Virol, 1991 Oct, 65(10), 5149 - 56
The herpes simplex virus 1 gene encoding a protease also contains within its coding domain the gene encoding the more abundant substrate; Liu FY et al.; The herpes simplex virus 1 open reading frames UL26 and UL26.5 are 3' coterminal . The larger, UL26 open reading frame encodes a protein approximately 80,000 in apparent molecular weight and contains the promoter and coding sequence of the UL26.5 gene, which specifies a capsid protein designated infected cell protein 35 . The larger product contains in its entirety the amino acid sequence of the smaller protein . We report that the UL26 gene encodes a protease which catalyzes its own cleavage and that of the more abundant product of UL26.5 . By inserting the coding sequence of an epitope to a cytomegalovirus monoclonal antibody and homologs of the immunoglobulin G binding domain of staphylococcal protein A into the 3' termini of the coding domains of the two open reading frames, we identified both products of the cleavage and determined that the cleavage site is approximately 20 amino acids from the carboxyl termini of both proteins.

J Infect Dis, 1991 Oct, 164(4), 711 - 9
Effects of staphylococcal toxic shock syndrome toxin 1 on aortic endothelial cells; Lee PK et al.; In staphylococcal toxic shock syndrome, hypotension and shock due to capillary leak may rapidly lead to death of the host . To investigate its pathogenesis, the cytotoxic effects of toxic shock syndrome toxin 1 (TSST-1) on porcine aortic endothelial cells (PAEC) were examined in vitro . TSST-1 killed PAEC (as measured by 51Cr release) in a dose- and time-dependent fashion and was blocked by anti-TSST-1 antibodies . Receptor-mediated endocytosis may be critical for the cytotoxic effects of TSST-1, as killing was inhibited by cold (4 degrees C) and by addition of chloroquine and methylamine . Furthermore, calcium and oxygen appeared necessary for TSST-1 effects on PAEC . Membrane receptor binding studies indicated PAEC bind TSST-1 with high affinity (Kd = 5.7 x 10(-7) M) and had 2.2 x 10(4) receptors/cell . Last, as measured by 125I-labeled albumin flux in a transendothelial permeability model, TSST-1 enhanced the permeability of PAEC monolayers in a dose- and time-dependent manner.

FEBS Lett, 1991 Sep 23, 290(1-2), 135 - 8
The Pro117 to glycine mutation of staphylococcal nuclease simplifies the unfolding-folding kinetics; Kuwajima K et al.; Kinetics of unfolding and refolding of a staphylococcal nuclease mutant, in which Pro117 is replaced by glycine, have been investigated by stopped-flow circular dichroism, and the results are compared with those for the wild-type protein . In contrast to the biphasic unfolding of the wild-type nuclease, the unfolding of the mutant is represented by a single-phase reaction, indicating that the biphasic unfolding for the wild-type protein is caused by cis-trans isomerization about the prolyl peptide bond in the native state . The proline mutation also simplifies the kinetic refolding . Importance of the results in elucidating the folding mechanism is discussed.

Biochemistry, 1991 Sep 17, 30(37), 8945 - 53
Effects of temperature on the fluorescence intensity and anisotropy decays of staphylococcal nuclease and the less stable nuclease-conA-SG28 mutant; Eftink MR et al.; Frequency-domain fluorescence spectroscopy was used to investigate the effects of temperature on the intensity and anisotropy decays of the single tryptophan residues of Staphylococcal nuclease A and its nuclease-conA-SG28 mutant . This mutant has the beta-turn forming hexapeptide, Ser-Gly-Asn-Gly-Ser-Pro, substituted for the pentapeptide Tyr-Lys-Gly-Gln-Pro at positions 27-31 . The intensity decays were analyzed in terms of a sum of exponentials and with Lorentzian distributions of decay times . The anisotropy decays were analyzed in terms of a sum of exponentials . Both the intensity and anisotropy decay parameters strongly depend on temperature near the thermal transitions of the proteins . Significant differences in the temperature stability of Staphylococcal nuclease and the mutant exist; these proteins show characteristic thermal transition temperatures (Tm) of 51 and 30 degrees C, respectively, at pH 7 . The temperature dependence of the intensity decay data are shown to be consistent with a two-state unfolding model . For both proteins, the longer rotational correlation time, due to overall rotational diffusion, decreases dramatically at the transition temperature, and the amplitude of the shorter correlation time increases, indicating increased segmental motions of the single tryptophan residue . The mutant protein appears to have a slightly larger overall rotational correlation time and to show slightly more segmental motion of its Trp than is the case for the wild-type protein.

J Immunol, 1991 Sep 15, 147(6), 1877 - 83
Human IgA and IgG F(ab')2 that bind to staphylococcal protein A belong to the VHIII subgroup; Sasso EH et al.; Staphylococcal protein A (SPA) is a bacterial membrane protein that possesses, in addition to its Fc gamma-binding activity, a distinct specificity for the Fab region of some IgM, IgA, IgG, and IgE . The Fab site that binds to SPA has been localized to the V region of the Ig H chain . In a previous study of human monoclonal and polyclonal IgM, we demonstrated that binding to SPA was highly restricted to molecules of the VHIII subgroup, and that nearly all VHIII IgM were able to bind SPA . The present study examines the VH composition of SPA-binding and SPA-nonbinding fractions of purified human polyclonal IgA, and IgG F(ab')2 fragments . We found that 22% of the IgA and 15% of the IgG F(ab')2 bound to SPA-agarose . Analysis with VH subgroup-specific antisera indicated that the SPA-binding fraction of IgA was dominated by the VHIII subgroup, and the SPA-binding fraction of IgG F(ab')2 contained only VHIII molecules . Furthermore, substantial portions of the total VHIII protein in IgA and in IgG F(ab')2 bound to SPA . We conclude that Fab binding to SPA is both restricted to and highly prevalent among human VHIII molecules, regardless of Ig class . These results suggest that protein A is an Ig superantigen.

Am J Ophthalmol, 1991 Sep 15, 112(3), 278 - 82
Anterior chamber aspirate cultures after uncomplicated cataract surgery; Dickey JB et al.; We cultured anterior chamber aspirates of 30 patients who had uncomplicated extracapsular cataract extraction or phacoemulsification . The aspirate was obtained at the time of wound closure . Of 30 patients, 13 (43%) had culture-positive anterior chamber aspirates . The total number of organisms recovered was 18, with three patients having multiple organisms identified . The most commonly isolated organisms were coagulase-negative Staphylococcus sp., occurring in eight of 18 isolates (44%) . No eyes in our study developed endophthalmitis, even though almost one half had viable organisms growing from the anterior chamber aspirates . Inoculum sizes were extremely small (10 to 20 colony-forming units/ml) . This study suggests that in humans, the anterior chamber is capable of clearing a low inoculum of bacteria after cataract surgery without the development of endophthalmitis.

Can J Microbiol, 1991 Sep, 37(9), 722 - 4
Comparison of the pathogenicity of three species of coagulase-negative Staphylococcus in a mouse model with and without a foreign body; Ferguson KP et al.; Staphylococcus schleiferi, Staphylococcus lugdunensis, and Staphylococcus epidermidis produce a high incidence of abscesses in a mouse model with an implanted foreign body . We investigated the significance of the foreign body in this process . Fourteen strains of S . schleiferi, S . epidermidis, and S . lugdunensis were tested in our model . A preadhered foreign body was implanted into one mouse group, followed by injection of a test strain . Another group received injection without implant . Abscesses were assessed at 7 days; foreign bodies and infected tissues were cultured . The percent of samples that developed abscesses or were culture positive was compared for each strain . Nearly all mice infected with S . schleiferi developed abscesses and were culture positive . The foreign body made no difference in abscess formation for three of four S . schleiferi but increased the incidence of both organism recovery and abscess for three of five S . epidermidis . The foreign body enhanced abscess formation for four of five S . lugdunensis, with all five strains yielding significantly more culture recovery . Although the pathogenicity of nine strains was increased by the foreign body, five strains yielded high abscess and culture recovery rates that were not enhanced by its presence.

Ophthalmology, 1991 Sep, 98(9), 1370 - 5
Effect of intravitreal dexamethasone on ocular histopathology in a rabbit model of endophthalmitis; Maxwell DP Jr et al.; The histologic and clinical effect of intravitreal dexamethasone was studied in a rabbit model of experimentally induced gentamicin sensitive Staphylococcal epidermidis endophthalmitis . The study compared four treatment groups: vitrectomy alone (group I), vitrectomy plus intravitreal gentamicin (group II), vitrectomy plus intravitreal gentamicin and dexamethasone (group III), and intravitreal gentamicin and dexamethasone without vitrectomy (group IV) . All eyes treated with dexamethasone, with or without vitrectomy, exhibited less clinical and histologic manifestations of inflammation . This study suggests that suppression of inflammation through the use of intravitreal dexamethasone leads to preservation of ocular architecture with potentially improved retinal function and visual benefit.

J Mol Biol, 1991 Sep 5, 221(1), 7 - 14
In a staphylococcal nuclease mutant the side-chain of a lysine replacing valine 66 is fully buried in the hydrophobic core; Stites WE et al.; The crystal structure of the staphylococcal nuclease mutant V66K, in which valine 66 is replaced by lysine, has been solved at 1.97 A resolution . Unlike lysine residues in previously reported protein structures, this residue appears to bury its side-chain in the hydrophobic core without salt bridging, hydrogen bonding or other forms of electrostatic stabilization . Solution studies of the free energy of denaturation, delta GH2O, show marked pH dependence and clearly indicate that the lysine residue must be deprotonated in the folded state . V66K is highly unstable at neutral pH but only modestly less stable than the wild-type protein at high pH . The pH dependence of stability for V66K, in combination with similar measurements for the wild-type protein, allowed determination of the pKa values of the lysine in both the denatured and native forms . The epsilon-amine of this residue has a pKa value in the denatured state of 10.2, but in the native state it must be 6.4 or lower . The epsilon-amine is thus deprotonated in the folded molecule . These values enabled an estimation of the epsilon-amine's relative change in free energy of solvation between solvent and the protein interior at 5.1 kcal/mol or greater . This implies that the value of the dielectric constant of the protein interior must be less than 12.8 . Lysine is usually found with the methylene groups of its side-chain partly buried but is nevertheless considered a hydrophilic surface residue . It would appear that the high pKa value of lysine, which gives it a positive charge at physiological pH, is the primary reason for its almost exclusive confinement to the surface proteins . When deprotonated, this amino acid type can be fully incorporated into the hydrophobic core.

Otolaryngol Head Neck Surg, 1991 Sep, 105(3), 419 - 21
Fine-needle aspiration of diffuse cervical lymphadenopathy in patients with acquired immunodeficiency syndrome; Shapiro AL et al.; Although diffuse cervical lymphadenopathy is one of the earliest and most common findings in patients with AIDS, the appropriate diagnostic approach in these patients has yet to be determined . Fine-needle aspiration (FNA) was performed on 26 patients with AIDS in order to evaluate the role of FNA in patients with diffuse cervical adenopathy . Specimens were sent for cytology, bacterial culture, fungal culture, and acid-fast smear and culture . Ten patients had positive findings, including toxoplasmosis, histoplasmosis, tuberculosis, atypical mycobacterium, and methicillin-resistant staphylococcal infection . All patients with either unilateral adenopathy or lymph nodes 3 cm or larger had positive aspirates . A statistically significant difference between patients with lymph nodes smaller than 2 cm and those with nodes larger than 2 cm was found . Fine-needle aspiration of a representative node in patients with AIDS may allow prompt diagnosis of diffuse lymphadenopathy . Rapid initiation of appropriate treatment can lead to symptomatic improvement . The need for excisional biopsy of involved lymph nodes may be obviated . Fine-needle aspiration is recommended as a diagnostic tool in selected patients with diffuse cervical lymphadenopathy and AIDS.

Ophthalmology, 1991 Sep, 98(9), 1376 - 8
Intralenticular infections; D'Mellow G et al.; Antecedent trauma resulting in endophthalmitis is not uncommon . However, primary intralenticular infection is a rare occurrence . Primary intralenticular fungal infection has not been previously reported . The authors present two cases: one of Paecilomyces infection and the other of Staphylococcus epidermidis infection limited to the crystalline lens . Both cases illustrate a delay in diagnosis.

J Thorac Imaging, 1991 Sep, 6(4), 60 - 4
Pulmonary disease in children with AIDS; Goodman PC; Children with acquired immunodeficiency syndrome (AIDS) may present with recurrent pneumonias or chronic debilitating illness . The chest radiographs of these patients demonstrate homogeneous densities representing staphylococcal or other pyogenic infections . Pneumocystis carinii pneumonia produces a diffuse, symmetric, fine-to-medium, reticulonodular pattern . Lymphocytic interstitial pneumonitis, a disease that is now an index diagnosis of AIDS in children under 13, may appear as a chronic, diffuse, small nodular infiltrate . An increasing number of pediatric AIDS patients will be observed in the future because of the large number of human immunodeficiency virus-infected women who are of childbearing age.

J Pediatr Surg, 1991 Sep, 26(9), 1058 - 63
Evaluation of mortality in surgical neonates over a 10-year period: nonpreventable, permissible, and preventable death; Hazebroek FW et al.; The mortality pattern and causes of death in surgical neonates were evaluated over the period of 1980 through 1990 in order to assess whether any deaths might have been prevented . In the study period 1,010 neonates were admitted, 104 of whom died (10%) . These 104 neonates formed our study population . The Clinical Classification System (CCS) and the Therapeutic Intervention Scoring System (TISS) were used to assess the severity of illness . All 104 nonsurvivors were seriously ill (CCS 3 and 4; TISS greater than 20) . The mortality pattern was classified in three groups . Group A (nonpreventable death) included 56 deaths, 48 of which were due to the underlying disease . The other 8 patients died of a nonpreventable complication of treatment . Eleven of the patients in group B (permissible death) had (multiple) congenital anomalies associated with chromosomal anomalies . Three of them were not treated and in the other 8 treatment was initially started but later withdrawn . In most of the other 14 patients further treatment was withdrawn because of serious postoperative complications or inoperable cardiac anomalies . Group C (preventable death) consisted of 23 patients . Six of 10 had irreversible brain damage due to prolonged hypoxemia . In the other 13 patients of group C, death was due to sepsis acquired in the postoperative period and treated inadequately . Staphylococcus epidermidis was the most frequent isolated pathogen (7/13).

Infect Control Hosp Epidemiol, 1991 Sep, 12(9), 544 - 8
Complications associated with central venous catheters inserted in critically ill neonates; Hruszkewycz V et al.; OBJECTIVE: To assess the incidence and spectrum of complications associated with central venous catheter (CVC) placement in the critically ill infant . DESIGN: A prospective study of all babies hospitalized in a neonatal intensive care unit (NICU) from January 1989 to December 1989 . Potential risk factors associated with infection were evaluated by a case-control comparison . SETTING: Conducted at a university-affiliated, tertiary care community hospital . PATIENTS: Neonates requiring intensive care and a central venous catheter . Controls consisted of noninfected babies . RESULTS: Of 263 critically ill neonates, only 13 (4.9%) required a CVC insertion . Seventeen CVCs were placed in these 13 neonates for a total duration of 600 days (median, 32 days/cannula) . Fifteen (88%) of these cannulas had one or more complications during its catheter life including dislodgement or leakage (53%), occlusion or thrombosis (47%), infections (29%), or minor bleeding (12%) . Five babies (29%) developed 6 episodes of bloodstream infection including 3 sporadic cases due to Staphylococcus epidermidis and a cluster of fungemia due to Malassezia furfur associated with lipid emulsion therapy . Infants with a CVC-associated infection were a younger gestational age (24 weeks versus 32 weeks, p = .04) and weighed less at birth (580 g versus 1285 g, p = .02) . The overall rate of bloodstream infection was one episode per 100 days of catheter use . CONCLUSIONS: CVCs may be lifesaving to a critically ill neonate, but complications occur frequently . Use must be restricted to infants in whom alternate delivery routes of intravenous therapy or support are otherwise unavailable.

Cardiovasc Intervent Radiol, 1991 Sep-Oct, 14(5), 307 - 10
Myocardial abscess resulting in a pseudoaneurysm: case report; Roberts JH et al.; This report describes the diagnostic workup of a case of a pseudoaneurysm of the heart resulting from Staphylococcal pericarditis and/or abscesses of the myocardium . The pericardial effusion and myocardial abscesses were detected on 2D echocardiography and computed tomography . The resulting pseudoaneurysm could be demonstrated on magnetic resonance imaging and color Doppler ultrasound, and confirmed by left ventriculography . Color-encoded Doppler ultrasound alone would have provided the definitive diagnosis of the aneurysm.

Br Heart J, 1991 Sep, 66(3), 244 - 5
Conservative surgery in multiple cusp involvement in tricuspid valve endocarditis; Anderson JR et al.; Tricuspid and mitral valve endocarditis caused by Staphylococcus epidermidis in a 57 year old previously healthy man with no history of drug abuse presented as bi-ventricular failure and multiple episodes of pulmonary emboli . He was treated for four weeks with intravenous antibiotics and had serial echocardiographic assessment of the vegetation on the tricuspid valve . This was followed by mitral valve replacement, local excision of vegetation from all the three cusps of the tricuspid valve, and autologous pericardial reconstruction of these cusps with functional assessment by perioperative transoesophageal echocardiography . Postoperative cardiac function was excellent and serial echocardiographic assessment confirmed satisfactory tricuspid valve function . This is believed to be the first recorded case in which autologous pericardial repair was used to reconstruct all the three cusps in a tricuspid valve after excision of vegetations.

Infect Immun, 1991 Sep, 59(9), 3337 - 9
Functional evidence that the Ser-195 residue of staphylococcal exfoliative toxin A is essential for biological activity; Prevost G et al.; The substitution of the serine 195 residue of staphylococcal exfoliative toxin A by a cysteine residue led to a biologically inactive protein . This result is consistent with the hypothesis that exfoliative toxin A could be a protease or a lipase . However, no protease or lipase activity was detected with the native toxin.

Infect Immun, 1991 Sep, 59(9), 2987 - 93
Staphylococcal exotoxins stimulate nitric oxide-dependent murine macrophage tumoricidal activity; Fast DJ et al.; The staphylococcal exotoxins toxic shock syndrome toxin 1 (TSST-1) and enterotoxin B were tested for their ability to stimulate murine peritoneal macrophages (PM) for tumoricidal activity . Both toxins were found to stimulate oil-elicited, gamma interferon-primed PM monolayers to kill nonadherent P815 tumor targets . The mechanism of killing of toxin-stimulated tumoricidal activity involved the production of nitric oxide, as nitrite could be demonstrated in culture fluids, and NG-monomethyl-L-arginine, an inhibitor of nitric oxide production, abrogated toxin-stimulated tumoricidal activity . TSST-1 stimulated the secretion of tumor necrosis factor by PM monolayers in the presence and absence of gamma interferon . The mechanism of toxin-stimulated tumoricidal activity was also determined to be independent of the production of reactive oxygen intermediates in that TSST-1 failed to stimulate H2O2 production by PM . These results demonstrate that the staphylococcal exotoxins are capable of stimulating macrophage production of nitric oxide for tumor cytotoxicity and suggest that the nitric oxide thus produced may subsequently play a role in the pathogenesis of the diseases caused by these toxins.

Proc Natl Acad Sci U S A, 1991 Sep 1, 88(17), 7533 - 7
Staphylococcal exotoxins deliver activation signals to human T-cell clones via major histocompatibility complex class II molecules; Spertini F et al.; We investigated whether staphylococcal exotoxins (SEs), in addition to their capacity to induce T-cell activation restricted by the T-cell receptor (TCR) beta-chain variable region, can deliver an activation signal to human T-cell clones through major histocompatibility complex (MHC) class II molecules . Eleven human T-cell clones (9 alpha beta TCR and 2 gamma delta TCR clones) of different antigenic specificities were tested for their capacity to proliferate in response to toxic shock syndrome toxin 1 (TSST-1) and two SEs, SEA and SEB . In the absence of accessory cells, only 4 alpha beta TCR clones were stimulated to proliferate, each by a single SE, and to mobilize intracellular free Ca2+ in response to that SE, events indicative of TCR engagement and, presumably, recognition restricted by the beta-chain variable region . In the presence of accessory cells, each of the 11 T-cell clones was stimulated to proliferate by any one of the three SEs tested . This apparently TCR-unrestricted SE-mediated polyclonal proliferation of T-cell clones occurred in the absence of an increase in intracellular free Ca2+ and was not dependent on the presence of MHC class II expression on accessory cells . In contrast, SE-mediated polyclonal proliferation did not occur in 3 alpha beta TCR clones derived from an MHC class II-deficient patient . Furthermore, all of the three SEs induced the proliferation of 4 natural-killer-cell clones, suggesting that expression of TCR/CD3 complex is not essential for SE-mediated polyclonal proliferation of activated lymphocytes . These results indicate that MHC class II molecules transduce activation signals to human T- and natural-killer-cell clones.

Infect Immun, 1991 Sep, 59(9), 2978 - 86
Biodegradable and biocompatible poly(DL-lactide-co-glycolide) microspheres as an adjuvant for staphylococcal enterotoxin B toxoid which enhances the level of toxin-neutralizing antibodies; Eldridge JH et al.; Microspheres composed of biocompatible, biodegradable poly(DL-lactide-co-glycolide) (DL-PLG) and staphylococcal enterotoxin B (SEB) toxoid were evaluated as a vaccine delivery system when subcutaneously injected into mice . As measured by circulating immunoglobulin G (IgG) antitoxin titers, the delivery of SEB toxoid via DL-PLG microspheres, 1 to 10 microns in diameter, induced an immune response which was approximately 500 times that seen with nonencapsulated toxoid . The kinetics, magnitude, and duration of the antitoxin response induced with microencapsulated toxoid were similar to those obtained when an equal toxoid dose was administered as an emulsion with complete Freund adjuvant . However, the microspheres did not induce the inflammation and granulomata formation seen with complete Freund adjuvant . The adjuvant activity of the microspheres was not dependent on the superantigenicity of SEB toxin and was equally effective at potentiating circulating IgG antitrinitrophenyl levels in response to microencapsulated trinitrophenyl-keyhole limpet hemocyanin . Empty DL-PLG microspheres were not mitogenic, and SEB toxoid injected as a mixture with empty DL-PLG microspheres was no more effective as an immunogen than toxoid alone . Antigen-containing microspheres 1 to 10 microns in diameter exhibited stronger adjuvant activity than those greater than 10 microns, which correlated with the delivery of the 1- to 10-microns, but not the greater than 10-microns, microspheres into the draining lymph nodes within macrophages . The antibody response induced through immunization with microencapsulated SEB toxoid was protective against the weight loss and splenic V beta 8+ T-cell expansion induced by intravenous toxin administration . These results show that DL-PLG microsphere vaccine delivery systems, which are composed of pharmaceutically acceptable components, possess a strong adjuvant activity for their encapsulated antigens.

Antimicrob Agents Chemother, 1991 Sep, 35(9), 1919 - 22
Susceptibility of Staphylococcus species and subspecies to teicoplanin; Bannerman TL et al.; Twenty-four Staphylococcus species and their subspecies were examined for their susceptibilities to teicoplanin by disk diffusion (30-micrograms disk) and agar dilution for the determination of MICs . Moderately susceptible and resistant clinical strains were further tested for their susceptibilities to oxacillin and vancomycin . Teicoplanin resistance was not observed in the reference strains of the various Staphylococcus species isolated from healthy volunteers or animals . However, the novobiocin-resistant species Staphylococcus saprophyticus, Staphylococcus cohnii, Staphylococcus xylosus, Staphylococcus arlettae, Staphylococcus kloosii, and Staphylococcus gallinarum were less susceptible to teicoplanin (MIC, 2 to 8 micrograms/ml) than most of the novobiocin-susceptible species were (MIC, 0.5 to 4 micrograms/ml) . Clinical isolates of coagulase-negative species were generally less susceptible to teicoplanin than were reference strains . Seven percent of the Staphylococcus epidermidis clinical strains were moderately susceptible (MIC, 16 micrograms/ml) to teicoplanin . Of these strains, 70% were oxacillin resistant . For Staphylococcus haemolyticus strains, 11% were resistant (MIC, greater than 16 micrograms/ml) and 21% were moderately susceptible to teicoplanin . Of these strains, 95% were oxacillin resistant, No strains of S . epidermidis or S . haemolyticus were intermediate or resistant to vancomycin . Teicoplanin appears to be less active in vitro against oxacillin-resistant S . haemolyticus . However, teicoplanin is an effective antimicrobial agent against many Staphylococcus species.

Immunology, 1991 Sep, 74(1), 99 - 106
IgA-mediated inhibition of human leucocyte function by interference with Fc gamma and C3b receptors; Saito K et al.; The inhibitory effects of IgA from human colostrum, and IgA1 and IgA2 from human serum on the chemiluminescence (CL) response and phagocytosis of polymorphonuclear leucocytes (PML) to Staphylococcus epidermidis and the CL response to formylmethionyl-leucyl-phenylalanine (FMLP) were studied . The dose-dependent inhibition of the luminol-mediated CL response of human PML to the bacteria was observed in the presence of more than 0.1 mg/ml IgA from both colostrum and serum . The preincubation of PML with a solution of IgA enhanced the suppressive effect of IgA on the cells . Removal of IgA from the reaction mixture after preincubation resulted in recovery, with time, of the response of PML to the bacteria . The bacteria treated with IgA did not give rise to any inhibition of the response . The CL response of PML to FMLP was not affected by the presence of IgA in the reaction mixture . The decrease of phagocytic activity of PML in the presence of IgA resulted in a decrease of NADPH oxidase activity of PML after stimulation with the bacteria as compared with the absence of IgA . The effect of IgA on the receptors of Fc and C3b (CR1) on the surface of PML was measured by monitoring erythrocyte-antibody (EA) or erythrocyte-antibody-complement (EAC) rosette formation and by direct and indirect immunofluorescence techniques using anti-CR1 antibody and Fc-specific antibodies . The presence of IgA in the reaction mixture led to a quantitative decrease in CR1 and the ability to bind IgG to the surface of PML.

Vopr Virusol, 1991 Sep-Oct, 36(5), 402 - 7
{Monoclonal anti-idiotypic antibodies simulating the biological effects of human alpha-interferons}; Nagieva FG et al.; The method of somatic hybridization was used to generate a panel of hybridomas producing monoclonal anti-idiotypic antibodies (mono-Ai-Ab) imitating biological effects of human alpha-interferons (hIF-alpha) . Induction of syngeneic anti-idiotypic antibodies in BALB/c mice was achieved with monoclonal antibodies (MCA) IF-39 capable of neutralizing three kinds of hIF-alpha (lymphoblastoid, leukocyte, genetic-engineering) . The screening of mono-Ai-Ab was done by determinations of antiviral activity (AV-activity) of supernatants from growing hybrid cell cultures caused by the cytopathic effect of 10-100 doses of mouse encephalomyocarditis virus (MEMC) by a micromethod in Vero cells grown in 96-well plates . Mono-Ai-Ab were found to neutralize MCA IF-39 and not to bind with immunosorbent of staphylococcal reagent containing protein A and BALB/c mouse immunoglobulins . It was shown that mono-Ai-Ab possessed AV-activity against MEMC and vesicular stomatitis viruses and were not inferior in this activity to commercial preparations of leukocyte IF-alpha . Mono-Ai-Ab had tissue species-specificity triggering the mechanism of AV-activity in human and simian cells as well as bovine kidney cells (MDVK line) imitating hIF-alpha in this effect.

Antibiot Khimioter, 1991 Sep, 36(9), 21 - 4
{Effectiveness of dioxidine in a model of Staphylococcal necrotic-suppurative encephalomeningitis (according to electron microscopy data}; Stebaeva LF et al.; Dioxidine efficacy was studied on a model of staphylococcal necrotic suppurative encephalomeningitis . There was a relationship of the development of disease signs to the value of the contaminating dose . Dioxidine was shown to have a therapeutic effect . Purulent inflammation in the brain either undevelop or was more limited . Examination of the changes in the ultrastructure of the nervous tissue also confirmed the efficacy of dioxidine . The treated mice had single inflammation infiltrations in the presence of the intact structure of the nervous tissue, the cell composition of the infiltrations changing in the direction of predominance of the mononuclear phagocytes . The phagocytes were in the state of hyperactivity and phagocytosis was of the completed nature . Secondary phagocytosis of the phagocytes and cell detritus was recorded.

J Biochem (Tokyo), 1991 Sep, 110(3), 381 - 7
Artificial cell adhesive proteins engineered by grafting the Arg-Gly-Asp cell recognition signal: factors modulating the cell adhesive activity of the grafted signal; Maeda T et al.; An artificial cell adhesive protein could be engineered by grafting the RGDS tetrapeptide, the core sequence of the major cell adhesive site of fibronectin, to a truncated form of Staphylococcal protein A (tSPA) via cassette mutagenesis of the tSPA expression vector pRIT2T {T . Maeda et al . (1989) J . Biol . Chem . 264, 15165-15168} . We synthesized a panel of tSPA derivatives grafted with various RGDS-containing oligopeptides to address the problem of how the cell adhesive activity of the resulting tSPA derivatives was affected by the length and amino acid sequence of the grafted oligopeptides and by the sites on tSPA where the extra oligopeptides were inserted . The results showed that (i) the amino acid residues flanking the RGDS core sequence played a key role in modulating the cell adhesive activity of the grafted RGDS signal; (ii) at least two sites on tSPA, each corresponding to on e of the two HindIII sites of pRIT2T, were competent in sustaining the cell adhesive activity of the grafted signal; and (iii) the divalent tSPA containing the RGDS signal at both sites was more active than monovalent derivatives containing only one signal at either site . These results provide a strategic basis for engineering of artificial cell adhesive proteins by grafting the RGDS signal.

Nutr Hosp, 1991 Sep-Oct, 6(5), 300 - 10
{The effect of nutrition on bone marrow transplantation}; Oloriz MR et al.; Malnutrition alone may provoke considerable immune alterations, mainly in the immunocellular function, and in particular in the T lymphocyte and macrophage-monocytic function . This gives rise to an increase in proneness to infections, which in patients subjected to bone marrow transplants, will be increased by the intense neutropenia suffered by them . This study shows the greater incidence of infection among badly nourished patients compared to nourished patients . It was also observed that short bouts of high temperature mainly occurred in a subgroup of patients with an acceptable nutritional level, which in our opinion was the cause for none of them developing the infectious illness . Finally, one of the germs most often found in these patients is Staphylococcus epidermis, and the most common method of entry is through the catheters used in the administration of medicines and artificial nutrition . Emphasis is placed on the role played by nursing in preventing this type of infection.

Zh Mikrobiol Epidemiol Immunobiol, 1991 Sep, (9), 68 - 70
{The course of experimental staphylococcal sepsis in opposite mouse strains and first-generation hybrids}; Averbakh MM et al.; The survival time and histological lesions of the kidneys, liver, heart, and lungs were studied in CBA/Sto, C3HA/Mv and F1 (C3HA/Mv x CBA/Sto) mice for 15 days after i.v . injections with S . aureus pathogenic strains CFU B-243 in doses of 10(9), 10(8) and 2.5 x 10(8) microbial cells . CBA/Sto mice were found relatively resistant and C3HA/Mv mice, susceptible to infection caused by different doses of S . aureus, this being associated with different morphological picture in the viscera . F1 hybrids were at least as susceptible to the infections as any of the parent strains, suggesting recessive inheritance of resistance to staphylococcal infection.

Pathol Biol (Paris), 1991 Sep, 39(7), 700 - 8
{Vancomycin in 1991: current status and perspectives}; Casetta A et al.; Vancomycin is a narrow-spectrum glycopeptide antibiotic which is primarily active against Gram-positive organisms . Bacterial resistance develops rarely due to its numerous modes of action . The mode of action of vancomycin involves the inhibition of peptidoglycan synthesis . Vancomycin forms a stoichiometric complex with the peptidoglycan precursor UDP-N-acetylmuramyl pentapeptide by forming hydrogen bonds . In patients with renal insufficiency vancomycin clearance is reduced and elimination half-life prolonged . Vancomycin is the drug of choice in the treatment of methicillin-resistant staphylococcal infections and in the treatment of Gram-positive endocarditis and has been used as alternative therapy in the treatment or prophylaxis of Gram-positive infections in penicillin-allergic patients.

Rev Infect Dis, 1991 Sep-Oct, 13 Suppl 10, S782 - 9
Perioperative antibiotic prophylaxis of wound and foreign body infections: microbial factors affecting efficacy; Waldvogel FA et al.; Numerous microbial factors are responsible for perioperative infections and influence the efficacy of antibiotic prophylaxis . These factors include the staphylococcal carrier state, bacterial adherence to a number of host proteins, the production of glycocalyx by sessile bacteria, and shifts in antibiotic resistance . A full understanding of the mechanisms involved will lead to further reductions in the number of postoperative infections . Unfortunately, the microbial factors affecting prophylaxis cannot be evaluated separately under clinical conditions; they are easier to study under circumstances whose bacteriologic features are well defined and in which the presence of foreign materials (e.g., sutures) greatly potentiates pathogenic mechanisms . Such circumstances exist, for example, in infections developing after "clean" surgery and in experimental models . Since even clean wounds are found to be contaminated when sampled carefully, the control of infection is more a quantitative than a qualitative problem . The critical period for the development of infection is short: an antibiotic course not exceeding 24 hours seems effective in preventing infection.

Neurochirurgia (Stuttg), 1991 Sep, 34(5), 160 - 2
{Abscess of the pyramidal apex}; Kehler U et al.; The authors report on one case of an abscess at the pyramidal apex . The 52-year old male patient presented with pareses of the fifth and seventh cranial nerves and hypacusis on the right side . After diagnostic procedures (CT-scan, carotid angiography), a tumor at the apex of the right pyramid was expected . During surgery a large encapsulated mass was found containing pus . A bacterial agent could not be isolated . The abscess bordered on the mucosal lining of the sphenoid sinus and on the cells of mastoid bone . The starting point of an abscess at the pyramidal apex is most commonly an otitis media, most frequently caused by staphylococcus . Sterile abscesses are seen in almost 20% . Of differential diagnosis on has to keep in mind other space-occupying lesions especially epidermoid or dermoid cysts.

Mol Gen Mikrobiol Virusol, 1991 Sep, (9), 28 - 30
{A simple method of purifying staphylococcal alpha-toxin and a study of its properties}; Beilbaeva ML et al.; The simple method is proposed for isolation and purification of staphylococcal alpha-toxin that permits one to obtain the homogeneous toxic protein with high activity . The time necessary for maximal toxin production at cultivation has been defined . The thermostability and interferonogenic characteristics of the obtained alpha-toxin were studied.

Ital J Gastroenterol, 1991 Sep-Oct, 23(7), 416 - 20
The spontaneous bacterial peritonitis in cirrhotic patients . To a new gold standard; Leandro G et al.; Of 282 consecutive ascites prospectively collected in 54 months, Spontaneous Bacterial Peritonitis (SBP) was diagnosed in 8.5% of the cases, "probable" SBP in 31.1%, Bacteriascites (BA) in 3.5% and Sterile Ascites (SA) (negative ascitic fluid culture with PMN less than 250/mm3) in 74.8% . Escherichia Coli (41.6%) and Staphylococcus Epidermidis (60%) were the most frequent pathogens isolated in patients with SBP and BA, respectively . With regards to in-hospital mortality, 18% of patients with BA and 50% with SBP died; the mortality seemed to be related to the degree of hepatic and renal damage, to a higher peripheral and ascitic WBC concentration and to a lower pH of ascitic fluid (FA) . When the comparative analysis was applied to the four groups of ascites, a different distribution of clinical signs and biohumoral parameters appeared . As a matter of fact, abdominal pain, fever and rebound tenderness resulted significantly more frequent in SBP and "probable" SBP . Furthermore, the mean values of peripheral and ascitic WBC concentration, of serum creatinine and of ALT were statistically higher in SBP and "probable" SBP than in SA and BA groups . The strict relationship, both symptomatologic and biochemical, between SA and BA on the one hand and between "probable" SBP and SBP on the other, prompted us to conclude that "probable" SBP and SBP represent different patterns of the same disease . Therefore, the subclassification in the four groups outlined above would not be in accordance with the clinical practice and could give rise to the physician's confusion and uncertainty.

J Hosp Infect, 1991 Sep, 19 Suppl B, 47 - 57
Reduction in the colonization of central venous cannulae by mupirocin; Hill RL et al.; In an in-vitro simulation of an intravascular cannula enclosed in a fibrin sheath, 0.03 mg1(-1) of mupirocin prevented significant colonization {greater than 15 colony forming units (cfu)} by two clinical isolates of Staphylococcus epidermidis and one each of S . saprophyticus, S . hominis and S . haemolyticus . This suggests that in vivo, where protein binding of mupirocin is 95-97%, 1 mg 1(-1) of mupirocin at the cannula surface would be required to prevent colonization . These results support the findings of our previously published prospective controlled trial, in which mupirocin applied to the insertion sites of 172 internal jugular cannulae reduced the rate of colonization of cannula tips to 5%, compared with 25% for the 186 controls (P less than 0.001) . Of the 46 colonized cannula tips from 110 control patients, the same species was isolated from the skin of the insertion site in 67% and from the lumen flush in only 15% . Analysed by patient, mupirocin reduced the proportion of patients with colonized tips from 17% to 3% after 24 h of infection, and from 35 to 10% after 48 h (P = 0.002) . The use of agar containing charcoal, as a mupirocin neutralizer, and the incubation of tip-culture plates flooded with the Oxford staphylococcus, gave no evidence of carry over of mupirocin onto cannulae removed from mupirocin-treated patients.

J Formos Med Assoc, 1991 Sep, 90(9), 886 - 92
{Application of staphylococcal protein A for immunodiagnosis of early toxoplasma infection}; Lin DB et al.; RH strain Toxoplasma gondii was used to inoculate mice and rats . Sera collected at intervals were assayed by the indirect fluorescent antibody test (IFAT) and the latex agglutination test (LAT) to monitor the time-course change in titers of anti-toxoplasma IgM and IgG antibodies . In addition, the above sera and some anti-toxoplasma seropositive human sera (with IFAT antibody titers greater than or equal to 1:16) were adsorbed with staphylococcal protein A (SpA) and assayed for changes in IgM and IgG antibody titers so as to evaluate the usefulness of SpA adsorption in detecting anti-toxoplasma IgM which shows up early in toxoplasma infections . Samples assayed included 262 sera from mice, 65 sera from 5 rats and 85 human sera . The results revealed that parasite specific IgM-IFAT antibodies were detectable in 40% of the sera from mice 3 days after infection . After SpA adsorption, however, the IgM-IFAT antibody could be detected in a few specimens as early as 2 days post infection . IgG-IFAT and LAT antibodies first appeared in the sera on the 5th day of infection . In mice inoculated with freeze-killed tachyzoites (immunized mice), IgG-IFAT and LAT antibodies remained high from 16-35 days after inoculation, whereas IgM-IFAT antibodies were undetectable . Even after SpA treatment, only about half of the 16-day samples showed IgM antibody titers and the other specimens still remained negative . In general, IgM antibody titers increased 2-6 fold after SpA adsorption while IgG antibodies were almost completely removed after the treatment with a residual IgG-IFAT titer of less than or equal to 1:4.(ABSTRACT TRUNCATED AT 250 WORDS)

Proc Natl Acad Sci U S A, 1991 Sep 1, 88(17), 7715 - 8
Cold denaturation and 2H2O stabilization of a staphylococcal nuclease mutant; Antonino LC et al.; Cold denaturation is now recognized as a general property of proteins but has been observed only under destabilizing conditions, such as moderate denaturant concentration or low pH . By destabilizing the protein using site-directed mutagenesis, we have observed cold denaturation at pH 7.0 in the absence of denaturants in a mutant of staphylococcal nuclease, which we call NCA S28G for a hybrid protein between staphylococcal nuclease and concanavalin A in which there is the point mutation Ser-28----Gly . The temperature of maximum stability (tmax) as determined by circular dichroism (CD) was 18.1 degrees C, and the midpoints of the thermal unfolding transitions (tm) were 0.6 degrees C and 30.0 degrees C . These values may be compared with the tm of 52.5 degrees C for wild-type staphylococcal nuclease, for which no cold denaturation was observed under these conditions . When the stability of the mutant was examined in 2H2O by NMR, CD, or fluorescence, a substantial increase in the amount of folded protein at the tmax was noted as well as a decrease in tmax, reflecting increased stability.

Lett Appl Microbiol, 1991 Sep, 13(3), 175 - 8
Protein A insensitive ELISA detection of staphylococcal enterotoxin B; Rogemond V et al.; A sandwich enzyme-linked immunosorbent assay to detect staphylococcal enterotoxin B was developed using rat monoclonal antibodies as capture antibodies and as a biotinylated conjugate . This test was sensitive, less than 1 ng/ml of enterotoxin B was detected and interference by protein A was prevented by the use of rat monoclonal antibodies of the IgG2a isotype which were insensitive to protein A even at concentrations greater than 1000 ng/ml.

J Immunol Methods, 1991 Aug 28, 142(1), 53 - 9
Immunoassay to measure staphylococcal protein A in the presence of murine immunoglobulins; Knicker SM et al.; Immunoassays designed to measure low concentrations of staphylococcal protein A are subject to varying degrees of interference by excess IgG . We have developed an enzyme-linked immunosorbent assay (ELISA) that overcomes this problem by analyzing IgG-containing protein A samples in solutions buffered at pH 3.5 . Under these carefully selected conditions, protein A is absorbed efficiently by solid-phase chicken anti-protein A antibodies, but protein A-IgG complexes are largely dissociated . The assay has a protein A detection limit of 0.1 ng/ml, and the response is unaffected by 0.25 mg/ml murine IgG . The method should be useful for determining protein A contamination levels in antibodies purified by affinity chromatography on immobilized protein A resins.

J Immunol, 1991 Aug 15, 147(4), 1398 - 405
Control of T cell responses to staphylococcal enterotoxins by stimulator cell MHC class II polymorphism; Yagi JJ et al.; The bacterial toxic mitogens or superantigens are a family of related proteins that elicit potent T cell proliferative responses . These responses require APC that express MHC class II proteins, but they are not MHC restricted and they do not depend on a processing step, presumably because these mitogens bind directly to MHC class II molecules . These mitogens stimulate T cells by interacting in an unknown way with the portion of the TCR encoded by certain V beta gene segments . In this paper, we explore the importance of MHC class II polymorphism in T cell responses to staphylococcal enterotoxins . We find that certain MHC molecules present SEB to V beta 8-bearing T cells far better than others . These data suggest that one route of host defence against bacterial toxic mitogens may be to alter MHC class II molecules so that stimulation is inhibited.

Proc Natl Acad Sci U S A, 1991 Aug 15, 88(16), 7228 - 32
Both alpha-helices along the major histocompatibility complex binding cleft are required for staphylococcal enterotoxin A function; Russell JK et al.; The superantigen staphylococcal enterotoxin A (SEA) requires interaction with class II major histocompatibility complex (MHC) molecules to activate T cells . We have previously used the synthetic peptide approach to establish one side of the hypothetical class II foreign-antigen binding cleft, alpha-helical region 65-85 of the beta chain, as a binding site involved in accessory cell presentation of SEA to T cells . To further characterize the structural basis for MHC-SEA interaction we have examined the role of the alpha-helical regions of the class II alpha and beta chains in SEA function . Using the synthetic peptide approach, we have found that both alpha-helical regions are required for SEA-induced proliferation . Their corresponding peptides directly bound SEA . Although the beta-chain peptides were able to inhibit SEA binding to human and mouse cells, the alpha-chain peptides were not . The data suggest that the alpha-helices along both sides of the hypothetical class II MHC molecule binding cleft are required for SEA-induced function, whereas the beta-chain alpha-helix is sufficient for SEA binding . A model of superantigen presentation is proposed wherein the MHC beta chain, possibly region 70-80, interacts with SEA region 1-45, whereas another region of SEA binds region 51-80 of the alpha chain.

J Biol Chem, 1991 Aug 15, 266(23), 15427 - 31
The interfaces of actin and Acanthamoeba actobindin . Identification of a new actin-binding motif; Vancompernolle K et al.; Actobindin is an 88-amino acid polypeptide, containing two almost identical repeated domains of 33 and 34 residues . Depending on the molar ratios in which they are mixed, actobindin binds either one or two actin molecules . We cross-linked actobindin and actin in the 1:1 complex, using the zero-length cross-linker 1-ethyl-3(3-dimethylaminopropyl)carbodiimide . The cross-linked peptides were purified after consecutive CNBr cleavage and trypsin and Staphylococcus protease V8 digestions, and the cross-linked side chains were identified by amino acid sequencing . Isopeptide linkages were formed between residues Glu-100 of actin and Lys-16 of actobindin . In addition, we found a connection between one or more of the acidic residues 1,2, or 3 of actin and Lys-16 and Lys-52 of actobindin . The cross-linked regions in actobindin contain Leu-Lys-His-Ala-Glu-Thr motifs, similar to sequences observed in several other actin-binding proteins.

FEMS Microbiol Lett, 1991 Aug 15, 66(3), 319 - 22
A chloramphenicol-streptomycin-resistance plasmid from a clinical strain of Staphylococcus sciuri and its structural relationships to other staphylococcal resistance plasmids; Schwarz S et al.; A 5.1-kb plasmid, designated pSCS12, isolated from a naturally occurring Staphylococcus sciuri conferred resistance to chloramphenicol (CmR) and streptomycin (SmR) . Restriction endonuclease analyses of pSCS12 revealed partial structural homologies to the CmR-plasmids pC221 from S . aureus and pSCS1 from S . intermedius, to the SmR-plasmids pSAI-1 from S . hyicus and pS194 from S . aureus, as well as to the CmR/SmR plasmid pSK68 from S . aureus . Southern-blot hybridization with specific CmR- and SmR-gene probes confirmed these similarities and allowed the mapping of the CmR- and SmR-determinants in the S . sciuri plasmid pSCS12 . These observations lead to the suggestion that CmR/SmR-plasmids, such as pSCS12, may have evolved from CmR- and SmR-plasmids by interplasmidic recombination.

J Mol Biol, 1991 Aug 5, 220(3), 771 - 8
Kinetic analysis of the acid and the alkaline unfolded states of staphylococcal nuclease; Chen HM et al.; Thermodynamic analysis by differential scanning calorimetry shows that the folding/unfolding transition of staphylococcal nuclease is consistent with the two-state process . Stopped-flow kinetic measurements, monitoring the Trp140 fluorescence and covering five decades in time (2 ms to 300 s), indicate that the unfolding from pH 7.0 to 3.1 is monophasic (time constant 1.15 s) and from pH 7.0 to 12.2 is biphasic (time constants: one less than 2 ms and the other 0.6 s) . However, the folding, either from pH 3.1 to 7.0 or from pH 12.2 to 7.0, is triphasic (time constants 150 ms, 850 ms and 30 s from acid, 90 ms, 565 ms and 33 s from alkaline) . A simple sequential model, which agrees with the above observations for acidic folding/unfolding is, D3 in equilibrium D2 in equilibrium D1 in equilibrium N . The three Ds denote three sub-states of the unfolded state and N denotes the native state . These sub-states of D have similar enthalpy and tryptophan fluorescence, and their equilibrium cannot be shifted by temperature changes . However, they are kinetically distinctive . Data do not favor alternative mechanisms assuming parallel transitions of the three Ds to N, or complexity of the N state, or parallel transitions of sub-states of N1, N2 and N3 to D . Other more complex, branched or cyclic, kinetics are not considered because of the lack of evidence, pH dependence of the unfolding kinetics suggests that the unfolding is triggered by protonation of 0.8(+/- 0.3) ionizable groups, with a pKa of 3.9 or by deprotonation of 1.6(+/- 0.4) ionizable groups with pKa values near 10.5 . Circular dichroisms indicate that these three D states retain nonrandom chain conformation . Possible role of these "chain conformation" in the protein folding is discussed.

Can J Microbiol, 1991 Aug, 37(8), 586 - 9
Immunological studies on staphylococcal enterotoxin D: production of murine monoclonal antibodies and immunopurification; Shinagawa K et al.; Eight murine monoclonal antibodies (MAbs) against staphylococcal enterotoxin D (SED) were obtained by fusion of myeloma cells with mouse spleen cells immunized with SED only or a combination of SED and either enterotoxin A (SEA) or enterotoxin E (SEE) . When only SED was used as an immunogen, six MAbs were specific for SED only, whereas one MAb was reactive with both SED and SEE when both SEs were used as immunogens . One MAb reacted with SEA, SED, and SEE when both SEA and SED were used as immunogens . A MAb with the highest reactivity to SED was used to prepare an immunosorbent for purification of SED by immunoaffinity chromatography . Approximately 70% of the partially purified SED was recovered in the eluate . The purified SED was electrophoretically and antigenically pure . Immunoaffinity chromatography proved useful in the purification of SED in terms of ease of purification, percent enterotoxin, and enterotoxin purity.

Can J Microbiol, 1991 Aug, 37(8), 581 - 5
Production and characterization of murine monoclonal antibodies against staphylococcal enterotoxins A and E; Shinagawa K et al.; Six murine monoclonal antibodies (MAbs) against staphylococcal enterotoxin A (SEA) and enterotoxin E (SEE) were prepared by fusion of myeloma cells with mouse spleen cells immunized with SEA and SEE . Of five MAbs to SEA tested, two MAbs were reactive with only SEA, whereas three were specific for both SEA and SEE . On the other hand, one MAb to SEE was found to be specific for only SEE . To study specificities of the combining sites of these MAbs, competitive binding assays with either SEA or SEE and horseradish peroxidase conjugated MAbs were performed using unconjugated MAbs as inhibitors . The results obtained in the assays suggest that different epitopes may be located on SEA and that some of them may be cross-reacting epitopes between SEA and SEE.

Zentralbl Veterinarmed B, 1991 Aug, 38(6), 468 - 72
Comparative studies on bacteriolytic properties of Staphylococcus chromogenes and Staphylococcus hyicus; Lammler C; Staphylococcus chromogenes and Staphylococcus hyicus showed bacteriolytic activities towards a Micrococcus luteus reference strain . This was demonstrated on tryptone soya agar containing M . luteus cells . Both bacteriolytic enzymes could be isolated by ionic exchange chromatography and subsequent gel filtration . The isolated bacteriolysin of S . chromogenes lysed the M . luteus reference culture, was heat inactivated by 95 degrees C and precipitated specifically with antiserum produced against the bacteriolysin of S . hyicus.

Liver, 1991 Aug, 11(4), 248 - 52
Incidence of spontaneous bacterial peritonitis in patients with ascites . Diagnostic value of white blood cell count and pH measurement in ascitic fluid; Storgaard JS et al.; During a 21-month period, 65 consecutive patients admitted with ascites were included in a prospective study of the incidence of spontaneous bacterial peritonitis, and paracentesis was performed on admission . The ascitic fluid was cultured, ascitic leucocytes were counted and pH was measured . Bacterial growth was found in five patients with chronic liver disease, who were diagnosed as having spontaneous bacterial peritonitis (SBP), since no intra-abdominal focus could be demonstrated . Thus, the incidence of SBP in this material was 7.7% (95% confidence limits: 2.5-17%) . SBP was caused by Escherichia coli (n = 3), coagulase negative staphylococcus (n = 1), and Bacteroides species (n = 1) . Abdominal tenderness, abnormal intestinal sounds, fever and hepatic encephalopathy were equally frequent in the group with SBP and in patients with sterile ascites . Infection was not anticipated in any of the patients with SBP . In contrast to several previous studies, neither ascites pH nor ascites leucocyte counts were any help in obtaining a rapid diagnosis . Survival time of patients with SBP was significantly shorter than of patients without SBP.

Nippon Kyobu Geka Gakkai Zasshi, 1991 Aug, 39(8), 1204 - 7
{Chronic hemorrhagic empyema developed in thirty three years after the right pneumonectomy--a case report}; Shinada J et al.; A 55-year-old man was admitted because of exertional dyspnea . He had the right pneumonectomy thirty three years ago . Chest X-ray showed the mediastinal shift to the left . And chest CT scan showed right intrathoracic mass . The bloody pleural effusion was aspirated (Hb 9.4 g/dl) and its examination revealed Staphylococcus epidermidis . We resected the empyema cavity . During the operation, massive bleeding was experienced (total 23200 ml) . Pathologically, micro blood vessels were marked in the organized hematomas and the pleura . Chronic hemorrhagic empyema is a specific type of chronic empyemas and it is dangerous to remove of the hematomas because of massive bleeding.

FEMS Microbiol Lett, 1991 Aug 1, 66(2), 229 - 31
Nucleotide sequence of the ethidium efflux gene from Escherichia coli; Purewal AS; The nucleotide sequence of the gene specifying the ethidium efflux system of Escherichia coli has been determined . The translated open reading frame has identified a membrane-bound polypeptide of 110 amino acids (11,960 Da) which shares 42% identity with a staphylococcal protein specifying resistance to ethidium.

J Pathol, 1991 Aug, 164(4), 307 - 14
Necrotizing myopathy in critically-ill patients; Helliwell TR et al.; Skeletal muscle wasting is commonly observed in critically-ill patients and has been attributed to catabolic fibre atrophy and to neuropathy . This study describes the occurrence of a necrotizing myopathy in 15 out of 31 critically-ill patients who had percutaneous biopsies taken from the tibialis anterior muscles . While most cases showed necrosis of isolated fibres, 5 of the 12 patients who had serial biopsies showed progressive necrosis of up to 95 per cent of the fibres . One other case showed infarction and one case had staphylococcal vasculitis . Atrophy of type 1 and/or type 2 fibres was documented by morphometry in 12 cases . Myoglobin-containing casts were demonstrated immunohistochemically in renal tubules on either biopsy or necropsy material in 5 out of 7 cases . The presence of muscle necrosis was a clinically unexpected finding which may contribute to weakness, complicate the interpretation of tissue biochemistry and energy balance studies, and potentiate renal failure . The necrosis is probably multifactorial in origin, with ischaemia and sepsis contributing factors.

J Appl Bacteriol, 1991 Aug, 71(2), 176 - 81
Relation of human serum antibody against Staphylococcus epidermidis cell surface polysaccharide detected by enzyme-linked immunosorbent assay to passive protection in the mouse; Ichiman Y et al.; A specific and rapid enzyme-linked immunosorbent assay (ELISA) has been applied for the detection of immunoglobulins to Staphylococcus epidermidis cell surface polysaccharides in human serum . Positive IgG, IgM and IgA titres of more than 1: 6400, 1: 1600 and 1: 400 were observed with this assay against passive protective human serum . However, IgG, IgM and IgA titres of less than 1: 400, 1: 100 and 1: 50 were shown in non-protective serum . When the cross-reactivity of passive protective human serum to homologous and heterologous cell surface polysaccharides was examined by inhibition test with ELISA, remarkable inhibition was shown with homologous cell surface polysaccharide, whereas no inhibition was observed with heterologous substances . According to these results, the quantitation of human serum antibody by the ELISA method against Staph . epidermidis cell surface polysaccharide was found to be significant for the demonstration of passive protective activities against Staph . epidermidis.

J Am Vet Med Assoc, 1991 Aug 1, 199(3), 362 - 3
Staphylococcus hyicus abortion in a sow; Onet GE et al.; Five aborted fetuses from a mature sixth-parity sow with severe greasy pig disease as a gilt, were submitted for diagnostic evaluation . Necropsy of the fetuses revealed serogelatinous edema in the SC connective tissue of the ventral abdominal region (especially around the umbilicus), exaggerated amounts of serohemorrhagic fluid in the abdominal, pleural, and pericardial cavities, and hemorrhagic kidneys, with diminished consistency . Staphylococcus hyicus was isolated in pure culture from liver, lung, kidney, and brain tissues and from abdominal, pleural, pericardial, and gastric fluids.

Immunology, 1991 Aug, 73(4), 433 - 7
Does staphylococcal enterotoxin B bind directly to murine T cells?
Qasim W, Kehoe MA, Robinson JH.
We have investigated the binding potential of staphylococcal enterotoxin B (SEB) to murine T cells using the induction of early activation events in Th1 and Th2 T-cell clones in the absence of antigen-presenting cells (APC) as indicators of direct interactions between SEB and the T-cell receptor (TcR) . We consistently found that concanavalin A (Con A) induced rises in intracellular free calcium as well as inositol phosphate accumulation in APC-free T-cell clones . However, SEB uniformly failed to induce either calcium fluxes or inositol phosphate turnover in Th1 and Th2 T-cell clones in the absence of APC . In addition, we have used proliferation assays to show that (i) T-cell clones prepulsed with SEB did not respond when APC were added, (ii) APC-independent T-cell clones responded to soluble anti-TcR antibodies but not to SEB in the absence of APC, and (iii) SEB coupled to Sepharose beads did not stimulate T-cell clones in the absence of APC . Taken together our results argue against SEB binding to the TcR without the participation of MHC class II molecules.

Int J Food Microbiol, 1991 Aug, 13(4), 265 - 72
ELISA screening of staphylococcal enterotoxins by means of a specially developed test kit; Kraatz-Wadsack G et al.; The development of a new test kit for the detection of staphylococcal enterotoxins is described . This polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) is designed for qualitative and quantitative rapid detection of toxin serotypes A, B, C (subgroup 1), D and E in one test system . Results are obtained in 60 min . The lower limit of detection is 0.5 ng/ml; the precision is 90%.

Eur J Immunol, 1991 Aug, 21(8), 1963 - 6
Clonal expansion precedes anergy and death of V beta 8+ peripheral T cells responding to staphylococcal enterotoxin B in vivo; MacDonald HR et al.; Staphylococcal enterotoxin B (SEB) selectively stimulates T cells bearing T cell receptor V beta 8 domains and hence provides a useful model to study immunity and tolerance in vivo . We show here that V beta 8+ T cells in both CD4+ and CD8+ subsets expand dramatically (fivefold) in lymphoid tissues of mice 2-4 days following injection with SEB . This initial clonal expansion, which is accompanied by a transient hyper-reactivity to SEB, is followed by a rapid decrease in V beta 8+ cells and a concomitant induction of specific non-responsiveness which persists for at least 30 days . Selective death of V beta 8+ cells occurs during this latter phase . Taken together, our data indicate that clonal expansion, anergy and death can occur as sequential stages of an immune response in vivo.

J Virol, 1991 Aug, 65(8), 4057 - 62
The antigen of hepatitis delta virus: examination of in vitro RNA-binding specificity; Chao M et al.; The only known protein of hepatitis delta virus (HDV), the delta antigen, is found both within virus particles and within the nucleus of the infected cell, where it has one or more roles essential for RNA genome replication . Others have demonstrated that the antigen has the ability, in vitro, to specifically bind HDV RNA species . We report a further examination of this phenomenon, using partially purified recombinant protein, expressed as a fusion with the staphylococcal protein A . From Northwestern (RNA-immunoblot) analyses with both complete and various subdomains of HDV genomic and antigenomic RNAs, we found that a necessary feature for specific binding was that the RNA be able to fold to some extent into the so-called rodlike structure; this structure is a predicted intramolecular partial base-pairing of the circular RNA, with about 70% of all bases involved, so as to produce an unbranched rodlike structure . Six different subregions of the HDV rodlike structure, three on the genomic RNA and three on its complement, the antigenomic RNA, were tested and found to be sufficient for antigen binding . However, features in addition to the rodlike structure may also be necessary for specific binding, because we found that a similar structure present in the RNA of the potato spindle tuber viroid did not allow binding.

Blood, 1991 Aug 1, 78(3), 830 - 7
Recombinant granulocyte-macrophage colony-stimulating factor after autologous bone marrow transplantation for relapsed non-Hodgkin's lymphoma: blood and bone marrow progenitor growth studies . A phase II Eastern Cooperative Oncology Group Trial; Lazarus HM et al.; Sixteen patients with relapsed non-Hodgkin's lymphoma underwent autologous bone marrow transplantation and infusion of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) . Treatment consisted of involved-field radiotherapy, cyclophosphamide 60 mg/kg/d intravenously (IV) for 2 days, and fractionated total body irradiation (1,200 cGy) . Autologous bone marrow was thawed and infused IV, followed 3 hours later by the first infusion of IV rhGM-CSF 11 micrograms/kg/d over 4 hours . Infusions of rhGM-CSF were continued daily until either both neutrophil count exceeded 1,500/microL and platelet count exceeded 50,000/microL, or until 30 days after marrow re-infusion . Toxicities encountered were mild and included fever, chills, hypertension, alopecia, rash, diarrhea, stomatitis, myalgias, and synovial (knee) effusions . Neutrophil recovery greater than 500/microL occurred a median of 14 days (range, 9 to 30 days) after marrow infusion, significantly earlier than in a comparable group of historic controls who recovered counts at a median time of 20 days (range, 12 to 51 days) (P = .00002) . Median time to self-sustaining platelet counts greater than 20,000/microL was 23.5 days (range, 12 to 100 days), comparable with the historic group (P = .38) . One bacteremia (central venous catheter exit site infection with Staphylococcus epidermidis) and one local infection (Giardia lamblia in stool) occurred . Patients received a median of 11.4 (range, 4.4 to 20.2) x 10(4) colony-forming unit granulocyte-macrophage (CFU-GM) progenitors per kg . Stem cell progenitors CFU-GM, CFU-granulocyte, erythroid, monocyte, megakaryocyte (CFU-GEMM), and burst-forming unit-erythroid (BFU-E) were detected in the bone marrow as early as 7 days after marrow re-infusion, and increased in proportion to peripheral blood counts, but by 30 to 60 days still remained much lower than before transplant . Neutrophils transiently decreased in 13 of 16 patients (median decrease, 42%) within 24 to 72 hours of discontinuing rhGM-CSF infusions . These data suggest that rhGM-CSF therapy enhances neutrophil recovery by forcing stem cells to produce mature elements at an enhanced rate but may not affect marrow stem cell and early progenitor population sizes.

J Am Osteopath Assoc, 1991 Aug, 91(8), 807 - 12
Red eyes and red rash with fever: an uncommon initial presentation for staphylococcal obturator abscess with adjacent ischial osteomyelitis; White BD; "Red eyes and red rash with fever" is a common pediatric complaint; however, it is an uncommon initial presentation for staphylococcal obturator abscess with adjacent ischial osteomyelitis . The case of a 13-year-old boy who was admitted to the hospital with conjunctivitis, erythematous maculopapular lesions, and fever and had this final diagnosis is reported . The differential diagnosis included various bacterial diseases, several viral illnesses, and rickettsial disease.

Zentralbl Bakteriol, 1991 Aug, 275(3), 374 - 81
Activation of mononuclear immune cells in response to staphylococcal lipoteichoic acid; Ohshima Y et al.; The goal of the present study was to evaluate the influence of staphylococcal lipoteichoic acid (LTA) on the activation of mononuclear immune cells . A murine tumor necrosis-like factor (TNF-like) was induced in the sera of CD-1 mice which had been primed with heat/formalin-inactivated Propionibacterium avidum KP-40 and subsequently exposed to LTA extracted from Staphylococcus saprophyticus strain S 1 . Monoclonal antibody against murine TNF (anti-TNF) significantly inhibited the cytostatic activity of mice sera against transformed L-929 cells . Freshly isolated lymphocytes did not display interleukin 2 (Il-2) receptors, but receptors were expressed on Con A incubated cells and in significantly higher numbers after coexposure to staphylococcal LTA in vitro . Since the induction of TNF (macrophages) and Il-2 receptors (lymphocytes) represent stimulation of the mononuclear immune system, staphylococcal LTA may be considered to be an immunomodifier.

Zentralbl Bakteriol, 1991 Aug, 275(3), 358 - 63
Adhesion of Staphylococcus saprophyticus to renal tubular epithelial cells is mediated by an N-acetyl-galactosamine-specific structure; Gatermann S et al.; S . saprophyticus CCM883 and 9325 were found to adhere to the tubular cell line LLC-PK1 . An ELISA technique was used to determine adherence of bacteria and inhibition of adherence by various carbohydrates . Only N-acetyl-galactosamine was found to significantly inhibit adhesion (p less than 0.001), which suggests that the surface component mediating adhesion recognizes structures on the target cell that contain this carbohydrate.

Immunol Lett, 1991 Aug, 29(3), 219 - 22
Stimulation of CD2-negative T cells by staphylococcal enterotoxin superantigens; Duan XC et al.; A minor fraction of CD3+ T cells lacks expression of the CD2 antigen, which is the target for an "alternative" T cell activation pathway . CD2-CD3+ T cells can be stimulated by anti-CD3 or anti-T cell receptor (TCR) antibodies, indicating that the CD3/TCR signal transduction pathway functions in the absence of cell surface CD2 . In the present study we have analyzed whether CD2-CD3+ T cells also respond to antigen stimulation . We show here that cloned CD2-negative T cells expressing the alpha/beta TCR are activated by one or several staphylococcal enterotoxin "superantigens" . Activation of CD2-CD3+ T cell clones by staphylococcal enterotoxins resulted in IL-2 production and/or proliferative activity, and was dependent on the presence of HLA class II-bearing feeder cells . These data demonstrate that T cells can recognize (and respond to) antigen in the absence of a functional CD2 molecule.

J Hosp Infect, 1991 Aug, 18(4), 293 - 9
Phage typing and phage induction in carrier and invasive Staphylococcus epidermidis isolates; Ferreiros CM et al.; Fifty-nine Staphylococcus epidermidis isolates (30 carrier and 29 invasive) were phage typed using the phage set developed by Pulverer . Poor results were obtained as only 24% of the invasive and 27% of the carrier isolates were typable, with no differences between these percentages (P greater than 0.05) . Isolation of lysogenic phages present in the isolates was attempted by three different methods, induction being satisfactory only by overnight incubation with 0.1 mg l-1 mitomycin C and detection with 2,3,5-triphenyl-tetrazolium chloride . All isolates produced phages, and the lytic activity of these phages was expressed on more than 50% of the isolates in all cases (except one phage which lysed 10% of the carrier isolates and 13.8% of the invasive isolates) . The percentages of isolates lysed in the two groups (carrier and invasive) were statistically different (P less than 0.05) only for phages derived from four isolates.

Biochem Biophys Res Commun, 1991 Jul 31, 178(2), 467 - 73
HLA-DR is a procoagulant; Chelladurai M et al.; Numerous incidents of thromboembolic complications have been documented in cancer patients and in recipients of mismatched organ transplants . Tumor procoagulants have also been implicated in the process of metastasis . Two protein bands of 35,000 and 28,000 daltons isolated from human ovarian carcinoma possessed procoagulant activity . The 35,000 dalton protein had an amino terminal sequence identical to that of the major histocompatibility antigen HLA-DR . Further, isolation of the protein using immunoaffinity column chromatography with monoclonal antibody to HLA-DR resulted in the isolation of procoagulant activity . The immunoaffinity purified protein enhanced thrombin generation in recalcified normal plasma approximately 20- fold . HLA-DR procoagulant activity was completely inhibited by Staphylococcal enterotoxin A . We propose that the procoagulant nature of HLA-DR may contribute to thrombotic disorders in several cancers and in association with graft rejection . The ability of enterotoxin A to inhibit this procoagulant may lead to development of future therapeutic strategies.

Biochemistry, 1991 Jul 30, 30(30), 7425 - 37
Conformation of an enzyme-bound substrate of staphylococcal nuclease as determined by NMR; Weber DJ et al.; The dinucleoside phosphodiester dTdA is a slow substrate of staphylococcal nuclease (kcat = 3.8 X 10(-3) s-1) that forms binary E-S and ternary E-M-S complexes with Ca2+, Mn2+, Co2+, and La3+ . The enzyme enhances the paramagnetic effects of Co2+ on 1/T1 and 1/T2 of the phosphorus and on 1/T1 of six proton resonances of dTdA, and these effects are abolished by binding of the competitive inhibitor 3',5'-pdTp . From paramagnetic effects of Co2+ on 1/T2 of phosphorus, koff of dTdA from the ternary E-Co(2+)-dTdA complex is greater than or equal to 4.8 X 10(4) s-1 and kon greater than or equal to 1.4 X 10(6) M-1 s-1, indicating the 1/T1 values to be in fast exchange . From paramagnetic effects of enzyme-bound Co2+ on 1/T1 of phosphorus and protons, with use of a correlation time of 1.6 ps on the basis of 1/T1 values at 250 and 600 MHz, 7 metal-nucleus distances and 9 lower-limit metal-nucleus distances are calculated . The long Co2+ to 31P distance of 4.1 +/- 0.9 A, which is intermediate between that expected for direct phosphoryl coordination (3.31 +/- 0.02 A) and a second sphere complex with an intervening water ligand (4.75 +/- 0.02 A), suggests either a distorted inner sphere complex or the rapid averaging of 18% inner sphere and 82% second sphere complexes and may explain the reduced catalytic activity with small dinucleotide substrates . Seventeen interproton distances and 108 lower limit interproton distances in dTdA in the ternary E-La(3+)-dTdA complex were determined by NOESY spectra at 50-, 100-, and 200-ms mixing times . While metal-substrate and interproton distances alone did not yield a unique structure, the combination of both sets of distances yielded a very narrow range of conformations for enzyme-bound dTdA, which was highly extended, with no base stacking, with high-anti glycosidic torsional angles for dT (64 degrees less than or equal to chi less than or equal to 73 degrees) and dA (66 degrees less than or equal to chi less than or equal to 68 degrees) and predominantly C-2'-endo sugar puckers for both nucleosides . Although the individual nucleosides are like those of B-DNA, their unstacked conformation, which is inappropriate for base pairing, as well as the conformational angles alpha and gamma of dA and zeta of dT, rule out B-DNA.(ABSTRACT TRUNCATED AT 400 WORDS)

J Clin Immunol, 1991 Jul, 11(4), 193 - 204
Characterization of T-cell subsets and T-cell receptor subgroups in pigtailed macaques using two- and three-color flow cytometry; Axberg I et al.; In order to characterize macaque T-lymphocyte subsets, we used a chromophore from a dinoflagellate, peridinin chlorophyll A protein (PerCP), which, like fluorescein isothiocyanate (FITC) and R-phycoerythrin (PE), can be excited by a 488-nm laser and emits light at 670 nm without spectral overlap with FITC and PE . Mouse monoclonal antibodies were conjugated with FITC, PE, and PerCP to detect CD4+ and CD8+ cells in macaque peripheral blood lymphocytes (PBL) subsets before and after activation and in nonactivated thymocytes . Resting and activated macaque blood CD4+ T-cells could be clearly delineated into discrete subsets with either CD28, CD45RA, or CD45RO as a second marker and CD26, CD29, CD44, or CD69 as a third marker . CD8+ cells were further subdivided by expression of similar combinations of markers . A subset of CD8+ CD28- T-cells in blood expressed the activation marker CD69, suggesting that they were already activated . Virtually all CD4+CD8+, CD4+CD8-, and CD4-CD8+ macaque thymocytes expressed CD2, CD3, and CD18 and not CD25, CD44, or CD45O, but macaque thymocyte subpopulations did differ in their expression of CD28 and CD29 . The expression of T-cell receptor (TCR) subgroups on macaque PBL and thymocytes was analyzed before and after activation with staphylococcal enterotoxins (superantigens) . The pattern of T-cell variable-region expression in macaques was similar to that seen in humans, with a high frequency of T cells expressing V beta 8 . After superantigen stimulation, only minor changes in TCR V beta expression were detectable in PBL . A dramatic increase in V beta 8 expression was seen after stimulation of macaque thymus with staphylococcal enterotoxin D (SE-D), a minor increase after toxic shock syndrome toxin 1 (TSST-1) stimulation, and a simultaneous decrease in V beta 6 levels.

Ann Otol Rhinol Laryngol, 1991 Jul, 100(7), 540 - 3
Toxic shock syndrome associated with pharyngitis and submandibular space abscess; Sales JH et al.; Toxic shock syndrome continues to be encountered more frequently with the head and neck areas as sources of the toxin . In head and neck surgery practice it is most commonly noted following nasal packing . An unusual case associated with staphylococcal pharyngitis and spontaneous submandibular space abscess is reported and the literature concerning the subject is reviewed . Treatment is eradication of the infective focus, aggressive support of vital functions, and parenteral antistaphylococcal antibiotics.

Med Trop (Mars), 1991 Jul-Sep, 51(3), 289 - 92
{Misleading forms of visceral leishmaniasis in children . Apropos of 5 cases}; Ben Becher S et al.; The authors report the cases of five children in whom kala-azar was undiagnosed at first instance . In these cases, the diagnosis was misled because of incomplete features (lack of fever, splenomegaly or hypergammaglobulinemia) an associated disease (hydatic cyst of the liver) or a complication dominating the clinical pattern (septicemia, staphylococcus respiratory infection) . In one case, the patient was explored in order to diagnose portal hypertension.

Antimicrob Agents Chemother, 1991 Jul, 35(7), 1298 - 302
Effects of intravitreal dexamethasone on concentration of intravitreal vancomycin in experimental methicillin-resistant Staphylococcus epidermidis endophthalmitis; Smith MA et al.; Intravitreal corticosteroids in the treatment of bacterial endophthalmitis remain controversial . We utilized an experimental rabbit model of methicillin-resistant Staphylococcus epidermidis endophthalmitis (i) to calculate the intravitreal vancomycin concentration in rabbit eyes receiving intravitreal vancomycin alone or in combination with intravitreal dexamethasone and (ii) to determine whether an intravitreal steroid has any effect on intravitreal vancomycin levels . All right eyes were infected and all left eyes were uninfected . The rabbits were divided into two treatment groups: (i) 32 eyes (group I) were injected with intravitreal vancomycin, 1.0 mg (0.1 ml); (ii) 32 additional eyes (group II) were injected with intravitreal dexamethasone, 400 micrograms (0.1 ml), in addition to vancomycin . Measurement of intravitreal vancomycin concentration was performed following sacrifice, utilizing a microbiologic agar diffusion assay . Analyses of intravitreal vancomycin concentrations were performed by using model-independent parameters, with area under the concentration-time curves derived by trapezoidal approximation . The intravitreal vancomycin concentration was significantly lower in both uninfected and infected group II eyes (P less than 0.002) . Analysis of intravitreal vancomycin concentration-time relationships was performed by using a nonlinear least-squares regression program; data best fit a one-compartment model . In addition, no vancomycin-dexamethasone interaction could be demonstrated . The reduced level of intravitreal vancomycin in the presence of intravitreal dexamethasone may have important clinical implications.

Protein Seq Data Anal, 1991 Jul, 4(1), 21 - 3
The N-terminal amino acid sequence of phenol hydroxylase contains a dinucleotide-binding sequence motif; Sejlitz T et al.; The N-terminal sequence of phenol hydroxylase from Trichosporon cutaneum was determined by Edman degradation of the integral protein and of fragments obtained by hydroxylamine cleavage and by digestion with Staphylococcus V8 protease . A continuous sequence of 80 residues from the N terminus was determined: TKYSESYCDV10, LIVGAGPAGL20 MAARVLSEYV30 RQKPDLKVRI40 IDKRSTKVYN50 GQADGLQCRT60 LESLKNLRLA70 DKIXSEXNDM80 . A single N-terminal sequence was detected, suggesting two identical subunits in the dimeric enzyme . We suggest the occurrence of an FAD-binding site near the N terminus . The C-terminal sequence is -LSTA, as determined by carboxypeptidase digestion.

Cell Immunol, 1991 Jul, 135(2), 372 - 82
The superantigen Pseudomonas exotoxin A requires additional functions from accessory cells for T lymphocyte proliferation; Legaard PK et al.; We have examined the functions required of accessory cells (AC) for murine thymocyte proliferation induced by Pseudomonas exotoxin A (PE) and have compared these functions to those required of a known superantigen, staphylococcal enterotoxin B (SEB) . We demonstrate that PE, like SEB, preferentially stimulates PNA+ thymocytes expressing a specific V beta element within the T cell receptor . However, PE requires functions from AC that are distinct from those required by SEB . AC treated with paraformaldehyde (PCHO) prior to stimulation supported thymocyte proliferation induced by SEB but not PE . However, when AC were treated with PCHO subsequent to stimulation with PE, thymocyte proliferation was observed, which suggests that PE requires antigen processing in addition to presentation . Furthermore, treatment of AC with lysosomotropic agents abrogated thymocyte proliferation induced by PE but not SEB . Antibodies to MHC class II molecules inhibited thymocyte proliferation induced by both PE and SEB . In addition, we observed that interleukin 1 alpha (IL-1 alpha) participated in the proliferation of thymocytes induced by PE but not SEB . Thus, our data indicate that PE is a unique microbial superantigen that requires additional AC functions for T lymphocyte proliferation.

JPEN J Parenter Enteral Nutr, 1991 Jul-Aug, 15(4), 412 - 6
A new approach to the diagnosis of central venous catheter sepsis; Bozzetti F et al.; One hundred forty-four cancer patients harboring a central venous catheter (CVC) were prospectively investigated to assess the relationship between hub culture, clinical assessment of sepsis before removal, and CVC sepsis . In 22 patients, the CVC was removed because of clinical assessment of catheter sepsis expressed by the staff prior to the removal . For each CVC removal, peripheral blood (qualitative method), hub, and CVC tip (quantitative method) cultures were performed . Clinical sepsis (disappearance of fever after CVC removal) was observed in 13 patients, microbiologic "sepsis" (identification of the same microorganisms on the CVC tip and in the peripheral blood) in seven patients, and clinical and/or microbiologic sepsis in 16 patients . Staphylococcus epidermidis was the microorganism most frequently identified . Hub culture was negative in 48% and positive for a low number and a high number of colonies in 35% and 17%, respectively . The predictive value of hub culture was 96% when testing negative and 8% and 37% (p = 3 x 10(-3)) when testing positive for a low and a high number of colonies, respectively . Predictive values of clinical assessment were 55% if positive and 97% if negative . Combining hub cultures and clinical assessment, the risk of sepsis varied from 2% with both evaluations negative to 89% in the case of positive clinical assessment associated with positive high-count hub . Inasmuch as the CVCs used have a disposable hub, it is possible to have an accurate diagnosis of CVC sepsis without removing the CVC.

Am J Vet Res, 1991 Jul, 52(7), 1061 - 4
Bacterial and mycoplasmal flora of the healthy camelid conjunctival sac; Gionfriddo JR et al.; Healthy conjunctival sacs of 88 animals of 3 species of captive camelids (Lama glama, Lama guanicoe, Lama pacos) and llama-guanaco hybrids were sampled for bacterial and mycoplasmal flora . Mycoplasmas were not isolated from any animal . Eleven genera of bacteria were isolated . The most frequent isolates were Staphylococcus epidermidis and Pseudomonas spp . Nine varieties of Pseudomonas were found, which represented at least 3 Pseudomonas species . Many of the bacterial isolates (especially the pseudomonads) are potential pathogens in the eyes of these camelids.

Diagn Microbiol Infect Dis, 1991 Jul-Aug, 14(4), 275 - 80
Reproducibility of Staphylococcus epidermidis plasmid profiles; Hartstein AI et al.; The plasmid profiles of six isolates of Staphylococcus epidermidis were repetitively evaluated over an 8-month period . Each isolate was subcultured and stored at three different temperatures (-70 degrees C, -20 degrees C, and room temperature) and plasmid DNA was prepared from each subculture at 0, 1, 4, and 8 months by two different methods of plasmid extraction {using mixed alkyltrimethylammonium bromide (ATAB) or Brij 58 and deoxycholate (modified Parisi)} . Plasmid DNA bands were lost from two isolates when subcultures were kept at room temperature . This plasmid loss was confirmed by repetitive extractions and electrophoresis, as well as by restriction endonuclease analysis of the ATAB preparations . Profiles were otherwise highly related to one another, with occasional exceptions being extra or missing plasmid DNA bands of high molecular size . The latter findings were not reproducible . Plasmid DNA extracted by the modified Parisi method was not reliably digested with restriction endonuclease enzymes . We conclude that the plasmid profiles of Staphylococcus epidermidis isolates are highly reproducible as long as isolates are stored at less than or equal to -20 degrees C . Minor discrepancies in the number of plasmid DNA bands of large molecular size may occur . These are resolvable by repetitive testing or restriction endonuclease analysis of ATAB-extracted plasmid DNA preparations.

Cornell Vet, 1991 Jul, 81(3), 239 - 43
Bone abscess in the mandible of a quarter horse gelding; Smyth GB; A 4-year-old Quarter horse gelding presented with a swelling in the soft tissues over the junction of the body and ramus of the left mandible . Radiography showed a well circumscribed lytic area within the mandible surrounded by sclerosis unassociated with any tooth . Aspiration of the lesion yielded pus . The abscess cavity was opened, curetted and lavaged . A Penrose drain was placed in the abscess cavity for 10 days . Bacteriological culture of the exudate and soft tissues from the abscess produced a few colonies of Staphylococcus intermedius . Histopathology showed chronic pyogenic infection . The horse recovered well and was normal 1 year after treatment.

Vet Microbiol, 1991 Jul, 28(2), 157 - 69
Susceptibility of various animals and cultured cells to exfoliative toxin produced by Staphylococcus hyicus subsp . hyicus; Sato H et al.; In piglets inoculated with partially purified exfoliative toxin (pp-shET) produced by Staphylococcus hyicus subsp . hyicus, exfoliation was observed at 12 h after injection . Chickens inoculated with the same dose of pp-shET also showed exfoliation within 30 min of injection . However, exfoliation was not demonstrated in mouse, rat, guinea pig, hamster, dog or cat inoculated with pp-shET until 24 h after injection . In cultured cell lines, especially L-929 and Hep-2, the rounding effect occurred after incubation with pp-shET for 1 h . The rounding effect was also seen in five other cultured cells (NCTC 2544, HeLa/S3, HmLu-1, CHO and BHK-21) 6-24 h after exposure to pp-shET . These round cells survived for 72 h after inoculation and formed a monolayer 24 h after changeover to a toxin-free medium . The rounding effect was observed in cells after the formation of the monolayer, but not before . It was suggested that the rounding effect was not caused by the increase in cyclic AMP in cells inoculated with pp-shET but by the cleavage of intracellular contacts.

Mol Pharmacol, 1991 Jul, 40(1), 28 - 35
Distribution of m2 muscarinic receptors in rat brain using antisera selective for m2 receptors; Li M et al.; The DNA fragment encoding the third intracellular loop of rat m2 muscarinic receptor was fused to the gene for staphylococcal Protein A . The resultant fusion protein, expressed in bacteria, was purified via IgG affinity chromatography and used as an antigen to raise a polyclonal antiserum . Chinese hamster ovary cells transfected with cDNA coding for a single muscarinic receptor subtype were used as tissue sources to screen antisera . The antiserum was shown to immunoprecipitate quantitatively (greater than 90%) m2 receptors but not to precipitate m1, m3, m4, or m5 receptors . Additionally, immunoprecipitation by m2 antiserum could be inhibited by protein containing the third intracellular loop of the m2 receptor . This selective m2 antiserum was then used to study the distribution and density of m2 receptors in rat brain and heart . In agreement with previous studies, m2 receptors were found to be abundant in heart and comprise at least 92% of the total muscarinic receptor density . Hindbrain, brain stem, and midbrain regions such as cerebellum (75%), pons/medulla (70%), and thalamus/hypothalamus (43%) are also enriched in m2 receptors . In contrast, forebrain regions contain markedly lower percentages of m2 receptors, with cortex expressing 20%, hippocampus 19%, striatum 12%, and olfactory tubercle 20% of the total receptor density . Although the density of m2 receptors expressed as a percentage of total varied considerably from brain region to brain region, the absolute density of these receptors appeared relatively uniform throughout the brain . This study demonstrates that a gene fusion system can be used for efficient antibody production . The use of similar fusion protein antisera directed against other subtypes of muscarinic receptors should prove useful in future studies on regulation, function, and structure of muscarinic receptors.

Rev Cubana Enferm, 1991 Jul-Dec, 7(2), 130 - 4
{Bacteriological examination of the skin of patients who are to undergo cardiac surgery}; Mancebo Hernandez S et al.; A study was carried out with patients who would undergo surgical intervention due to congenital heart malformations; skin samples were taken for bacteriological cultures in the preoperative period, both in the ward and the operating room, in order to determine if there are differences and if pathogenic organisms are present . The paper points out a high positiveness to negative coagulase staphylococcus and discusses the possible pathogenicity.

Rev Cubana Enferm, 1991 Jul-Dec, 7(2), 121 - 9
{The risk of sepsis induced via the deep venous approach}; Gabella Rodriguez C et al.; In order to determine the sepsis induced by the practice of canalizing the deep venous sector in revascularized patients, 43 catheters removed from 43 patients were cultured; no sepsis induced by this practice was found and 13 catheters had positive cultures, which amounted to 30.3% of the sample under study; five catheters (38.4%) with positive cultures belonged to patients who showed no sepsis; eight catheters (61.5%) with positive cultures corresponded to patients who showed septic focci . The average stay was 4.07 days . The germ most frequently found was negative coagulase staphylococcus.

Ann Hematol, 1991 Jul, 63(1), 49 - 53
Functional characterization of canine lymphocyte subsets; Hotzl C et al.; Functional characterization of subsets of T lymphocytes is essential for transplantation studies in dogs, as it is in other species . We studied the function of T cells separated by two mouse monoclonal antibodies recognizing complementary subsets--an antibody directed to canine T cells (MdT-P1) with an up-regulating function, and an antibody directed to human CD 8 (MT811) that cross-reacts with down-regulating canine T cells . Immunorosetting with sheep red blood cells and Percoll gradient allowed us to study depleted and enriched fractions . Their function was tested in mixed lymphocyte culture (MLC), cell-mediated cytotoxicity (CML), and coculture with B cells in a hemolytic plaque assay (PFC) . In MLC, MdT-P1-positive cells showed a high proliferative response, and MT811-positive cells responded poorly to allogeneic cells . Vice versa, MT811- negative cells responded strongly, and MdT-P1-negative cells were poor responders but strong stimulators . Effector cells of CML were separated following 8 days of culture and prior to mixing with target cells . Enriched and depleted fractions with either antibody showed low cytotoxic activity as compared with unseparated cells . When added to unseparated effector cells MT 811-positive cells suppressed cytotoxicity . B cells were obtained by rosetting with staphylococcal protein A (SPA) . Their immunoglobulin production was studied following 6 days of culture stimulated by pokeweed mitogen in a reverse hemolytic plaque assay . Again, MT 811-positive cells added to the culture suppressed, and MT 811-negative cells enhanced immunoglobulin production . In conclusion, immunorosetting with two monoclonal antibodies allowed us to distinguish subpopulations of canine T cells with up-regulating (helper/inducer) from those with down-regulating (suppressor) activity.

J Infect Dis, 1991 Jul, 164(1), 108 - 13
Effect of subinhibitory concentrations of clindamycin and trospectomycin on the adherence of Staphylococcus epidermidis in an in vitro model of vascular catheter colonization; Khardori N et al.; Septicemia, often due to Staphylococcus epidermidis, is a life-threatening complication associated with indwelling vascular catheters . An important factor in the development of such infections is glycocalix, or slime . An in vitro model that mimics intravenous delivery systems in humans was developed . It consisted of a modified Robbins device containing slices of silicone catheters in the removable ports, through which S . epidermidis diluted in 5% dextrose-normal saline with 10% heat-inactivated normal human serum was run, with and without clindamycin and trospectomycin . S . epidermidis was recovered from all catheters in the absence of antibiotics; no growth was detected with antibiotics . Scanning electron microscopy demonstrated significant reduction in glycocalix and no visible organisms with all concentrations except 0.5 micrograms/ml trospectomycin and 1 microgram/ml clindamycin; for those, a moderate amount of glycocalix and a few bacteria were seen . Thus, subinhibitory levels of trospectomycin and clindamycin may have a role in the prevention of microbial adherence to vascular catheters.

Arch Ital Anat Embriol, 1991 Jul-Sep, 96(3), 185 - 99
{Experimental models for studying the effects induced by staphylococcal toxins A and B on human keratinocytes in culture}; Arco A et al.; The toxic effects of the two serotypes of staphylococcal exotoxin: exfoliatin A (ETA) and B (ETB) on two experimental models: organotypic cultures of the skin and cellular cultures of the epidermis reconstructed "in vitro" have been studied . The results show that, in both cases, purified ETB (Fig . 4a, 6-6a, b, c) reproduces the characteristics of Lyell's Staphylococcal Syndrome that is the intraepidermal cleavage either between the granulosa and the spinous layers or at the granulosa layer of the epidermis . As the images of the LM and EM demonstrate, purified ETA behaves differently in the two experimental models; in fact, it produces the same effect as purified toxin B on the organotypic cultures (Fig . 3a, b), whereas it causes no alterations in the epithelial cultures reproduced "in vitro" (Fig . 5a, b).

Vopr Virusol, 1991 Jul-Aug, 36(4), 306 - 9
{Monoclonal antibodies to natural human gamma-type interferon}; Nagieva FG et al.; Nine hybridomas producing monoclonal antibodies (MCA) to natural human gamma interferon (IF-gamma) were generated . BALB/c mice were immunized with nonpurified IF-gamma preparation synthesized by lymphoid cells of the peripheral blood of donors in response to induction with staphylococcal enterotoxin A . For the first time somatic hybridization was done with the use of a medium with a high content of HEPES which maintained hybridomas viable for a long period of time . Out of 9 hybridomas, two (-gamma 6.1 and gamma-8.1) were shown to produce MCA with a high binding activity in the enzyme immunoassay . The same MCA effectively neutralized the biological activity of natural IF-gamma.

Biull Eksp Biol Med, 1991 Jul, 112(7), 73 - 6
{Production of monoclonal antibodies to staphylococcal alpha-toxin using the technique of in vitro immunization}; Beliaev NN et al.; The aim of this study is production of monoclonal antibodies (MAB) to staphylococcal alpha-toxin (SAT) . SAT was obtained from culture medium of S . aureus s . 0-15 with ion-exchange chromatography and chromatofocusing . SAT was conjugated with CH-sepharose 4B and used for in vitro immunization of spleen cells, extracted from intact BALB/c mouse.

Kardiologiia, 1991 Jul, 31(7), 66 - 8
{Limitation of the cardiac depressive effect caused by various forms of staphylococcal infection by using dalargin, a synthetic analog of endogenous opioids}; Pashutin SB et al.; Male Wistar rats were used to examine the cardiodepressive action of experimentally induced generalized infection and bacterial shock and the ability of the synthetic opioid dalargin to enhance resistance the damaging effect of bacterial intoxication . The cardiac performance of the infected rats was studied in a model of an isolated perfused heart with the working left ventricle . In the rats, generalized Staphylococcus infection was ascertained to result in a dose-dependent inhibition of heart performance at all study stages, showing the peak of cardiodepressive action on days 3 of postinfection . The experimental animals given dalagin displayed a high infection resistance, as manifested predominantly by prevented profound cardiac disorders due to systemic administration of dalargin or by a positive inotropic effect of the drug supplemented to perfusate, the effect being particularly pronounced in bacterial shock or in early generalized infection.

Mikrobiyol Bul, 1991 Jul, 25(3), 238 - 46
{Neonatal sepsis}; Senses DA et al.; We noticed coagulase positive staphylococcus is the most common pathogen in the neonatal sepsis in our neonatology unit . We followed 22 cases with neonatal sepsis . Blood cultures revealed coagulase staphylococcus in 9 cases, coagulase negative staphylococcus in 6 cases, Pseudomonas spp . in 5 cases, E . coli in 1 case, Klebsiella spp . in 1 case . The most common symptom was apnea . CRP positivity was noted in 15 cases (68.2%) while increase in immature/total neutrophil ratio was observed in 12 cases (54.5%) . However, we detected leukopenia in 2 cases (9.1%) . Also, the birthweight of the cases died due to neonatal sepsis were below 2000 gr.

Mol Gen Genet, 1991 Jul, 227(3), 377 - 84
Organization, promoter analysis and transcriptional regulation of the Staphylococcus xylosus xylose utilization operon; Sizemore C et al.; The Staphylococcus xylosus xyl genes were cloned in Staphylococcus carnosus by complementation to xylose utilization . Xylose isomerase assays under inducing (xylose present) and non-inducing (xylose absent) conditions indicated the presence of a regulated xylA gene on the recombinant plasmid . The nucleotide sequence (4520 bases) revealed three open reading frames with the same polarity . They were identified by sequence homologies as xylR, encoding the Xyl repressor, xylA, encoding xylose isomerase and xylB, encoding xylulokinase . Primer extension analyses indicated constitutive transcription of xylR and xylose-inducible transcription of xylA . Promoter consensus sequences were found upstream of both transcriptional start sites . A transcriptional terminator between xylR and xylA separates the different transcriptional units . Potential regulatory elements were identified by sequence analysis and suggest a repressor-operator mechanism for the regulation of xylAB expression.

J Appl Bacteriol, 1991 Jul, 71(1), 72 - 7
Surface characteristics and adhesion of Escherichia coli and Staphylococcus epidermidis; Gilbert P et al.; Surface hydrophobicity, surface electrokinetic potential and the ability to adhere to nitric-acid cleansed glass surfaces has been assessed throughout the growth, in batch culture, of Escherichia coli and Staphylococcus epidermidis . In both instances adhesiveness and surface hydrophobicity decreased in early- to mid-exponential phase . Cell surface charge, on the other hand became more electro-negative for E . coli but electro-neutral for Staph . epidermidis as the cells proceeded to divide . Adhesiveness correlated directly with surface electronegativity and hydrophobicity for Staph . epidermidis but inversely with surface electro-negativity for E . coli.

J Clin Microbiol, 1991 Jul, 29(7), 1493 - 7
Molecular epidemiology of Staphylococcus haemolyticus strains isolated in an Albanian hospital; Renaud F et al.; A recent outbreak of erythroderma in young children in an Albanian hospital was investigated . The etiology was not established, but Staphylococcus haemolyticus was frequently isolated from the affected children and from staff working in the same unit . Possible relationships among the isolates were investigated by using classical techniques (biotype, antimicrobial susceptibility, and extrachromosomal DNA pattern) and by restriction endonuclease analysis (REA) of total DNA . Control isolates of proven pathogenicity from hospitalized patients in Lyon, France were subjected to the same procedures . Distinct REA patterns were obtained after digestion with two enzymes in 7 of 10 isolates from five affected children . Six distinct patterns were observed in nine isolates from six staff members; two REA patterns from patient isolates and two from staff members were identical, and these were distinguishable by the other markers examined . Only two different REA patterns were found in the pathogenic control isolates despite the use of a third additional enzyme . Again, the isolates with the same REA patterns could be distinguished by their plasmid profile or antimicrobial resistance profile . REA of total DNA used in combination with other markers indicated that the Albanian isolates differed considerably, whereas the French pathogenic isolates showed little variability.

Biotechnology (N Y), 1991 Jul, 9(7), 642 - 6
Engineering proteins to enhance their partition coefficients in aqueous two-phase systems; Kohler K et al.; We describe a novel method to partition recombinant proteins into the polymer-rich top phase in poly(ethylene glycol) (PEG)4000/potassium phosphate aqueous two-phase systems . The concept is based on fusion of a gene fragment encoding a short peptide sequence to the product gene of interest thereby changing the partitioning properties of the expressed product protein as a fusion to the peptide . The model protein in this study, ZZ, is a two domain molecule based on staphylococcal protein A (SPA) which distributes evenly in PEG/salt systems . A tetrapeptide sequence, AlaTrpTrpPro (designated the partitioning peptide), was designed by molecular modeling techniques to include exposed tryptophan residues and to have a coding DNA sequence which is possible to polymerize in an obligate head-to-tail fashion at the DNA level . Gene fragments encoding one and three partitioning peptides, respectively, were fused to the 3' end of the ZZ gene and the fusion proteins were produced intracellularly in Escherichia coli . The partition coefficients of ZZ proteins containing zero, one and three fused partitioning peptides were determined in three PEG 4000/potassium phosphate aqueous two-phase systems of different compositions . In all three phase systems, there were dramatic effects on the partition coefficient by the fused partitioning peptides . In the phase system with the largest effects, the partition coefficient was enhanced from 1.6 to 11.6 by fusing one tetrapeptide sequence to the 147 amino acid model ZZ protein . By the fusion of three partitioning peptides, the coefficient was increased to 96.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemistry, 1991 Jun 25, 30(25), 6103 - 14
Interactions of the acid and base catalysts on staphylococcal nuclease as studied in a double mutant; Weber DJ et al.; The mechanism of the phosphodiesterase reaction catalyzed by staphylococcal nuclease is believed to involve concerted general acid-base catalysis by Arg-87 and Glu-43 . The mutual interactions of Arg-87 and Glu-43 were investigated by comparing kinetic and thermodynamic properties of the single mutant enzymes E43S (Glu-43 to Ser) and R87G (Arg-87 to Gly) with those of the double mutant, E43S + R87G, in which both the basic and acidic functions have been inactivated . Denaturation studies with guanidinium chloride, CD, and 600-MHz 1D and 2D proton NMR spectra, indicate all enzyme forms to be predominantly folded in absence of the denaturant and reveal small antagonistic effects of the E43S and R87G mutations on the stability and structure of the wild-type enzyme . The free energies of binding of the divalent cation activator Ca2+, the inhibitor Mn2+, and the substrate analogue 3',5'-pdTp show simple additive effects of the two mutations in the double mutant, indicating that Arg-87 and Glu-43 act independently to facilitate the binding of divalent cations and of 3',5'-pdTP by the wild-type enzyme . The free energies of binding of the substrate, 5'-pdTdA, both in binary E-S and in active ternary E-Ca(2+)-S complexes, show synergistic effects of the two mutations, suggesting that Arg-87 and Glu-43 interact anticooperatively in binding the substrate, possibly straining the substrate by 1.6 kcal/mol in the wild-type enzyme . The large free energy barriers to Vmax introduced by the R87G mutation (delta G1 = 6.5 kcal/mol) and by the E43S mutation (delta G2 = 5.0 kcal/mol) are partially additive in the double mutant (delta G1+2 = 8.1 kcal/mol) . These partially additive effects on Vmax are most simply explained by a cooperative component to transition state binding by Arg-87 and Glu-43 of -3.4 kcal/mol . The combination of anticooperative, cooperative, and noncooperative effects of Arg-87 and Glu-43 together lower the kinetic barrier to catalysis by 8.1 kcal/mol.

J Steroid Biochem Mol Biol, 1991 Jun, 38(6), 695 - 701
17 beta estradiol affects the expression of guinea pig blood leukocyte MHC antigens; Debout C et al.; Sex hormones have an effect on various immune responses but the mechanisms of action are unknown . One of these mechanisms might be a modification of expression of major histocompatibility complex (MHC) antigens in blood leucocytes . Estradiol-induced variations of the expression of guinea pig blood leukocytes MHC antigens (GPL-A) was studied . Class I and class II MHC antigens were detected by a sensitive rosetting method using specific alloimmune sera (AIS) and staphylococcal protein A-coated sheep red blood cells (SPA-SRBC) and evaluated by counting the number of bound SPA-SRBC per 100 cells . MHC antigens decreased after estrogen treatment . Estradiol modifies the expression of GPL-A antigens on the mononuclear cells including the Kurloff cells, which are involved in immunity or in a natural killer effect, but did not affect the expression of polymorphonuclear cells, ones which are not involved in immunity.

Am J Respir Cell Mol Biol, 1991 Jun, 4(6), 489 - 96
Identification of pneumocin, a developmentally regulated apical membrane glycoprotein in rat lung type II and Clara cells; Lwebuga-Mukasa JS; Pneumocin (Mr, 165 kD) is a recently identified apical membrane surface sialoglycoprotein marker of type II pneumocytes . A murine monoclonal IgG1 subclass-producing clone 4A (4A mAb), which was developed against the purified pneumocin, and recognized pneumocin on Western blots of adult rat lung homogenates, was used to study expression of the glycoprotein in developing rat lungs . Pneumocin localized to apical membranes of late fetal, neonatal, and adult rat type II pneumocytes as well as Clara cells in situ, by immunofluorescence and immunoelectron microscopy . Faint immunofluorescence was observed in 17-d fetal lungs . However, 19-d fetal lungs showed intense immunofluorescence with the antibody . On immunoelectron microscopy, apical membranes of 19-d fetal and adult rat lung type II cells were labeled by 4A mAb, but type I cells were not stained . On Western blots, amounts of pneumocin increased up to the fourth day after birth, when near-adult levels were attained . Lower molecular weight forms (Mr, 80 to 90 kD) were recognized in 17-d fetal lung . These bands decreased in amount with a corresponding increase in the 165-kD band that was typically observed in adult lungs . Immunoglobulins that were eluted from polyvinylidene difluoride strips containing the 165-kD band recognized the Mr 80 to 90 kD bands and 50-kD component, suggesting that fetal forms of the protein shared an epitope in common with the adult pneumocin . Reactivity of the glycoprotein with 4A mAb was destroyed by enzymatic digestion with trypsin and staphylococcal V8 protease . These data demonstrate that pneumocin is a developmentally regulated apical membrane marker of differentiated type II and Clara cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Epidemiol Infect, 1991 Jun, 106(3), 497 - 505
A large outbreak of food poisoning of unknown aetiology associated with Stilton cheese; Maguire HC et al.; Between November 1988 and January 1989, a total of 155 people in 36 reported outbreaks suffered gastrointestinal symptoms associated with eating Stilton cheese, produced from unpasteurized cow's milk in the English midlands . Symptoms were suggestive of a staphylococcal illness but extensive laboratory testing of cheeses implicated in several of the outbreaks failed to detect any pathogen, toxin or chemical . Control measures were implemented, and included a voluntary withdrawal of the implicated Stilton cheese from sale on 23 January 1989 and a subsequent decision to use pasteurized milk in production of the cheese.

Neurology, 1991 Jun, 41(6), 828 - 30
Chronic inflammatory demyelinating polyradiculoneuropathy of childhood: treatment with high-dose intravenous immunoglobulin; Vedanarayanan VV et al.; We treated four children with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) with high-dose intravenous immunoglobulin (IVIG) . All patients received 400 mg/kg of IVIG a day for 5 days during relapses, and one patient received additional periodic infusions of 400 mg/kg . All patients showed excellent recovery of motor strength following each relapse that was treated with IVIG . Compared with plasmapheresis (which was used to treat relapses earlier), recovery of function with IVIG treatments was similar, and in two patients it was superior, to plasmapheresis . There were no side effects with IVIG treatments as compared with plasmapheresis with which two children had infection of central lines with Staphylococcus epidermidis, one had profuse bleeding from accidental extrusion of a central line, and one had multiple episodes of major venous thromboses . High-dose IVIG was a safe and effective adjunctive therapy for childhood CIDP in these four patients.

Am J Med, 1991 Jun, 90(6), 758 - 62
Native valve Staphylococcus epidermidis endocarditis: report of seven cases and review of the literature; Arber N et al.; This report describes seven patients from three university hospitals whose native valve infective endocarditis was caused by Staphylococcus epidermidis . The literature on endocarditis caused by S . epidermidis is also reviewed and the clinical features of patients with native valve endocarditis due to this organism are compared with those of patients from a general series of infective endocarditis cases . Compared with infective endocarditis caused by other organisms, S . epidermidis endocarditis tends to occur more frequently in male patients . Patients with S . epidermidis endocarditis exhibit fewer embolic complications and skin manifestations . The frequency of congestive heart failure is lower in this group . The relative indolent course and apparent rarity of native valve S . epidermidis endocarditis necessitate a high index of suspicion for early diagnosis.

J Clin Invest, 1991 Jun, 87(6), 1925 - 35
Staphylococcus epidermidis induces complement activation, tumor necrosis factor and interleukin-1, a shock-like state and tissue injury in rabbits without endotoxemia . Comparison to Escherichia coli; Wakabayashi G et al.; Tumor necrosis factor (TNF) and IL-1 are thought to mediate many of the pathophysiologic changes of endotoxemia and Gram-negative bacteremia . In these studies, heat-killed Staphylococcus epidermidis were infused into rabbits to determine whether an endotoxin (LPS)-free microorganism also elicits cytokinemia and the physiologic abnormalities seen in Gram-negative bacteremia . S . epidermidis induced complement activation, circulating TNF and IL-1, and hypotension to the same degree as did one-twentieth the number of heat-killed Escherichia coli . Circulating IL-1 beta levels had a greater correlation coefficient (r = 0.81, P less than 0.001) with the degree of hypotension than TNF levels (r = 0.48, P less than 0.02) . Leukopenia, thrombocytopenia, diffuse pulmonary capillary aggregation of neutrophils, and hepatic necrosis with neutrophil infiltration were observed to the same extent after either S . epidermidis or E . coli infusion . However, S . epidermidis infusion did not induce significant (less than 60 pg/ml) endotoxemia, whereas E . coli infusion resulted in high (11,000 pg/ml) serum endotoxin levels . S . epidermidis, E . coli, LPS, or S . epidermidis-derived lipoteichoic acid (LTA) induced TNF and IL-1 from blood mononuclear cells in vitro . E . coli organisms and LPS were at least 100-fold more potent than S . epidermidis or LTA . Thus, a shock-like state with similar levels of complement activation as well as circulating levels of IL-1 and TNF were observed following either S . epidermidis or E . coli . These data provide further evidence that host factors such as IL-1 and TNF are common mediators of the septic shock syndrome regardless of the organism.

Dis Colon Rectum, 1991 Jun, 34(6), 502 - 5
Toxic shock syndrome after pilonidal cystectomy . Report of a case; Shlasko E et al.; Toxic shock syndrome is an uncommon disease associated with staphylococcal infections . Although most frequently reported in menstruating women and associated with tampon use, toxic shock syndrome has been described following many types of surgical procedures . In this report we describe a case of toxic shock syndrome occurring in a previously healthy young man after elective surgery for a pilonidal cyst.

J Neurosurg, 1991 Jun, 74(6), 878 - 86
Pyogenic osteomyelitis of the spine in the elderly; Cahill DW et al.; The authors report 10 cases of spontaneous pyogenic spinal osteomyelitis encountered within a 3-year period . There were six women and four men, ranging in age from 60 to 84 years . Six cases occurred at the thoracic level, three at the lumbar level, and one in the cervical spine . No patient was diabetic, immunocompromised, or receiving steroid therapy, and none had a history of endocarditis or intravenous drug abuse . No patient had undergone previous spinal surgery . There were no instances of coexisting tuberculosis or malignancy . Contemporaneous cases with known predisposing factors have been excluded from this report; however, three patients did have a recent history of somatic infection, one with known sepsis . All 10 patients had been previously misdiagnosed, frequently by neurosurgeons and orthopedists as well as by internists and family practitioners . Three had undergone inappropriate or unnecessary surgical procedures, and two had received inappropriate radiation therapy . Seven cases were caused by Staphylococcus species . Gram-negative bacteria, or anaerobic infections . In the other three, no bacteriological diagnosis was made, secondary to prolonged antibiotic therapy before surgery . Each patient had developed symptomatic neural element compression, spinal instability, or both by the time of their referral . The patients with subcervical pyogenic spinal osteomyelitis underwent transthoracic or retroperitoneal decompression and corpectomy with simultaneous autologous bone grafting, followed by 6 weeks of bed rest and 6 weeks of intravenous broad-spectrum or organism-specific antibiotic therapy . They were then mobilized in orthoses for an additional 6 weeks . In no case were foreign implants employed or further stabilization procedures necessitated . One patient required an additional 6 weeks of antibiotics for recalcitrant Pseudomonas colonization . Despite the patients' advanced age and the extensive surgical procedures, there was no mortality and no neurological morbidity . All patients were asymptomatic or demonstrated objective improvement upon discharge from the hospital . In this subset of patients with spontaneous pyogenic vertebral osteomyelitis, the only predisposing factor was advanced age.

J Pediatr Surg, 1991 Jun, 26(6), 714 - 7
Spontaneous, isolated intestinal perforations in neonates with birth weight less than 1,000 g not associated with necrotizing enterocolitis; Meyer CL et al.; From January 1986 through December 1988, we have seen 7 cases of isolated intestinal perforation in 250 infants with birth weights less than 1,000 g (3% incidence) without histological or clinical evidence of necrotizing enterocolitis (NEC) . Patients had a mean birth weight of 670 g, gestational age of 25.1 weeks, and sustained a perforation at a chronological age of 10.4 days . No infants had been fed . A definite, blue-discolored abdomen was the only consistent clinical sign (n = 7) . Free intraperitoneal air on radiograms was rarely observed (n = 1) . Abdominal ultrasounds (n = 3) and metrizamide contrast studies (n = 3) were not diagnostic . The presence of an umbilical artery catheter (7/7), falling hematocrit (6/7), thrombocytopenia (5/7), and a positive diagnostic paracentesis were most commonly found . In 6 of 7 patients, this perforation was associated with coagulase-negative staphylococcal sepsis . Surgical or histological diagnosis showed focal perforation in either the terminal ileum (n = 4) or the transverse and descending colon (n = 3) . Survival was 3 of 7; 2 patients died of intracranial hemorrhage and 2 died of Candida sepsis . We conclude that (1) intestinal perforation can occur in the absence of NEC; (2) bluish discoloration of the abdomen is the most reliable clinical finding; and (3) perforation may be associated with coagulase-negative staphylococcal infection.

Biokhimiia, 1991 Jun, 56(6), 1057 - 68
{Study of rabbit IgG, modified with carboxycarbonic anhydride of diethylenetriaminopentacetic acid}; Kirillova NM et al.; The affinity-purified by chromatography on immobilized antigen rabbit IgG was modified with mixed carboxycarbonic anhydride of DTPA which markedly alters the interaction of charged residues in the protein molecule . To study the correlation between the antigen binding activity and the conformational mobility of IgG, the reactivity of modified IgG towards conformational probes targeted at variable and constant IgG domains, was investigated . The antibody against CH2 domains of IgG, staphylococcal protein A and protein antigen ferritin were used as conformational probes . It was found that modification of IgG amino groups entails the global increase in conformational mobility involving the Fab fragments, CH2 and, probably, the CH3 domains of the Fc portion of IgG . Taking advantage of Fab fragments modification it was shown that two processes contribute to the global increase in the conformational mobility of IgG . These processes are: i) stimulation of segmental flexibility and, ii) increase in the mobility within the Fv domains of the Fab fragments.

Pediatr Dermatol, 1991 Jun, 8(2), 147 - 51
Diffuse cutaneous mastocytosis mimicking staphylococcal scalded-skin syndrome: report of three cases; Oranje AP et al.; Three cases of diffuse cutaneous mastocytosis (DCM) were at first incorrectly diagnosed as staphylococcal scalded-skin syndrome . In the first patient, at age 1 day the disease was recognized promptly by simple techniques such as Darier's sign and Tzanck smear . Much delay in making the diagnosis occurred in the other two patients, however: almost 1 year and 15 years, respectively . Bullous manifestations in mastocytosis occur only in the first two or three years . In the first months the disease can be dangerous and life threatening . To distinguish mastocytosis from vesicular and bullous neonatal disorders, one should perform Darier's sign and a Tzanck smear . The diagnosis is confirmed by histopathologic studies . Treatment of the bullous manifestations is symptomatic, with zinc oxide paste and oral antihistamines, which may provide some relief . In addition, cimetidine and sodium cromoglycate may be beneficial . At a later age psoralen plus ultraviolet A therapy may also relieve the symptoms . Particular foods and medicines can liberate histamine and should be restricted as much as possible in extremely affected patients . Special care should be taken when these patients are to undergo anesthesia . The risk of complications during and after anesthesia is also present in other forms of mastocytosis.

Am J Dis Child, 1991 Jun, 145(6), 675 - 80
Factors associated with umbilical catheter-related sepsis in neonates; Landers S et al.; To determine factors associated with risk for umbilical catheter-related sepsis, we studied neonates with one or more catheters in place for more than 3 days . Among 225 infants with 357 umbilical catheters, catheter-related sepsis occurred in 14 infants (6%) . Catheter-related sepsis occurred in 5% of infants with umbilical arterial catheters and in 3% of infants with umbilical venous catheters . Staphylococcal species accounted for 71% of cases of catheter-related sepsis . Multiple logistic regression analysis revealed that very low birth weight and longer duration of antibiotic therapy were significantly associated with risk for umbilical arterial catheter-related sepsis . Increased risk for umbilical venous catheter-related sepsis was best predicted by the simultaneous occurrence of higher birth weight and infusion of hyperalimentation solution . Catheter duration correlated with duration of antibiotic therapy and with infusion of hyperalimentation solution for both types of catheters; however, in the multivariable analysis, duration of catheterization was not found to be a significant independent predictor of risk for catheter-related sepsis for either type of catheter.

Antibiot Khimioter, 1991 Jun, 36(6), 43 - 4
{The antibiotic prophylaxis of postoperative complications in breast cancer patients}; Shamilov AK et al.; The studies showed that infectious complications in patients operated for breast cancer (BC) most frequently developed after removal of the drainage tube resulting in poorer discharge favourable for development of infections . The causative agents in such cases are usually exogenous bacteria differing from endogenous ones (Staphylococcus epidermidis) inhabiting the human skin and sometimes contaminating the operative field . The endogenous bacteria are detectable bacteriologically in 60 per cent of the cases . Still, since the operation wound contains humoral and tissue immunity factors (specifically active against the host microflora) such bacteria rarely grow on artificial media (18 per cent) and even more rarely cause infections . Therefore, to prevent postoperative infections in patients with BC it should be recognized rational to use broad-spectrum antibacterial drugs such as ampiox, ampicillin, doxycycline, cephalosporins of the 2nd and 3rd generations, etc . for 5 to 6 days after the drainage removal . If an infection develops the preventive therapy should be replaced by an adequate therapy in accordance with the pathogen sensitivity.

Zh Mikrobiol Epidemiol Immunobiol, 1991 Jun, (6), 73 - 5
{Changes in the lymphocyte membrane lipids during immunization and staphylococcal infection}; Afonina GB et al.; The relationship between the state of the lipid complex of lymphocyte membranes and their functional activity has been studied in normal mice after immunization and in mice with staphylococcal infection . The study has revealed essential differences in the dynamics of free-radical processes under normal conditions and in the presence of pathology, as well as their relationship to changes in the functional state of lymphocytes and, in particular, to the development of immunodeficiency.

Vaccine, 1991 Jun, 9(6), 443 - 50
High antibody responses in rabbits immunized with influenza virus ISCOMs containing a repeated sequence of the Plasmodium falciparum antigen Pf155/RESA; Sjolander A et al.; Immunostimulating complexes (ISCOMs) are spherical structures where immunogens are presented as multimers in a matrix of the adjuvant Quil A . ISCOMs have been shown to enhance the immunogenicity of several antigens important to both human and veterinary vaccine development . We have coupled a fusion protein, designated ZZ-M2, comprising eight copies of the C-terminal repeat subunit EENV of the Plasmodium falciparum blood-stage antigen Pf155/RESA and two IgG-binding domains of staphylococcal protein A (SpA), to preformed influenza virus envelope protein ISCOMs . Rabbits immunized with the conjugated ISCOMs produced high titres of antibodies even after the first injection . These antibodies reacted with the EENV repeat sequence in ELISA and with Pf155/RESA in immunofluorescence on infected erythrocytes . The antibody response, which was sustained for more than 20 weeks, was efficiently boosted and superior or equal to that obtained after immunization with ZZ-M2 in Freund's complete adjuvant . In contrast, the antibody response induced in rabbits immunized with ZZ-M2 in Syntex Adjuvant Formulation-MF (SAF-MF) was weak and of short duration . The antibodies produced after immunization with ZZ-M2 coupled to influenza virus ISCOMs mainly recognized epitopes formed by two or more EENV subunits and were highly specific for Pf155/RESA . Furthermore, the antibodies efficiently inhibited merozoite reinvasion of erythrocytes in vitro, indicating that they recognized epitopes exposed on the native antigen . In addition, the ZZ-M2-conjugated ISCOMs also induced high titres of antibodies reacting with SpA or the influenza virus envelope protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Biotechnol Appl Biochem, 1991 Jun, 13(3), 412 - 21
Purification and characterization of recombinant human insulin-like growth factor II (IGF-II) expressed as a secreted fusion protein in Escherichia coli; Wadensten H et al.; Human insulin-like growth factor II (IGF-II) was produced in an Escherichia coli ompT strain as a 22.5-kDa fusion protein . IGF-II was fused to the carboxy-terminal of a synthetic 15-kDa IgG-binding protein, originating from staphylococcal protein A, via a unique methionine linker . During fermentation, the fusion protein was exported to the growth medium at levels exceeding 900 mg/liter and subsequently affinity purified on IgG Sepharose followed by ion exchange on S Sepharose . After chemical cleavage with CNBr, yielding an authentic IGF-II molecule, the recombinant IGF-II was purified to homogeneity by a two step procedure involving ion-exchange and reverse-phase HPLC . A substantial fraction of the secreted protein was found to be biologically active, eliminating the need for complex refolding procedures . The yield of highly purified and biologically active IGF-II was 5-7 mg/liter of fermenter broth . The IGF-II produced by this method displayed biochemical, immunological, receptor binding, and biological activity properties equal to those of native IGF-II isolated from human serum.

Clin Nephrol, 1991 Jun, 35(6), 243 - 51
Crescentic glomerulonephritis in Wegener's granulomatosis: morphology, therapy, outcome; Grotz W et al.; Fourteen patients with Wegener's granulomatosis (WG) and severe renal and extrarenal involvement were studied (serum creatinine on admission 5.8 +/- 3.4 mg/dl) . Renal histology showed a necrotizing, crescentic glomerulonephritis in all patients . Despite advanced renal disease on admission cyclophosphamide, steroids (in 13 patients) and plasma exchange (in 9 patients) caused a rapid and sustained improvement of renal function . Four patients required intermittent hemodialysis over a period of one week . After 2 weeks of treatment serum creatinine values below 2 mg/dl (n = 4) indicated a nearly complete recovery of renal function in the long-term follow up (mean serum creatinine achieved after 12 months therapy: 1.1 +/- 0.1 mg/dl (n = 4) . Therefore serum creatinine values observed after 2 weeks of therapy, appear to be of prognostic value with regard to renal outcome . No relapse of active WG or progressive renal deterioration was observed during follow-up (22 +/- 13 months) except in one patient with persisting renal impairment . Three patients died (staphylococcus sepsis, intracerebral hemorrhage during hypertensive crisis, pulmonary embolism) during the first two months of therapy . The decline of serum creatinine seemed to be a better indicator of successful therapy than the decrease of anticytoplasmatic antibody (ANCA), erythrocyte sedimentation rate (ESR) and hematuria . On admission ANCA titer neither correlated with serum creatinine, the degree of renal involvement, nor was it of prognostic value . ANCA, serum creatinine and hematuria normalized within 2 to 8 months, whereas ESR and proteinuria remained elevated . Our data indicate a good prognosis of WG even with advanced renal involvement and generalized vasculitis provided aggressive treatment is performed early.

J Am Acad Dermatol, 1991 Jun, 24(6 Pt 1), 970 - 2
Staphylococcal sepsis in HIV antibody seropositive psoriasis patients; Jaffe D et al.; The cases of three HIV-positive men with generalized psoriasis and staphylococcal sepsis are reported . In each case the skin appeared to be the source of infection . While the patients received antibiotic therapy, the psoriatic plaques resolved despite minimal or no topical treatment.

J Clin Microbiol, 1991 Jun, 29(6), 1221 - 4
Characterization of Staphylococcus hyicus with the ATB 32 Staph system and with conventional tests; Lammler C; The ATB 32 Staph system correctly identified 45 of 54 Staphylococcus hyicus cultures isolated from pigs and cattle . The biochemical profiles of the remaining nine cultures were not listed in the product data base . The 40 porcine and 14 bovine cultures resulted in three and seven different biochemical profiles, respectively . In parallel experiments, almost all S . hyicus cultures showed hemolytic reactions on chocolate agar, had CAMP-like reactivities in the zone of lysis of the staphylococcal beta-lysin, and had bacteriolytic properties on Micrococcus luteus cells . In addition, the S . hyicus cultures were unpigmented, were DNase positive, expressed an S . hyicus-specific teichoic acid, and were usually coagulase positive in porcine plasma . Most of the porcine strains were protein A positive, and two porcine cultures were clumping factor positive.

Rev Esp Enferm Dig, 1991 Jun, 79(6), 433 - 5
{Spontaneous rupture of the abdominal wall in cirrhotic patients with ascites}; Castellote J et al.; The abdominal wall hernia is a common finding in cirrhotic patients . Spontaneous disruption of the abdominal wall through these herniae is an uncommon complication, is associated with a high mortality and should be considered a gastroenterologic emergency . It occurs through a skin ulceration over the hernia in the 70% of all cases . Intravascular expansion plus long antibiotic prophylaxis with anti-staphylococcal agents are the mainstays of medical therapy . The definitive treatment must be surgical . Each case must be evaluated individually to determine the optimal surgical management, not necessarily on an emergent basis . We report three new cases surviving this complication.

Zentralbl Bakteriol, 1991 Jun, 275(2), 156 - 61
Visualization of endo-beta-N-acetylglucosaminidase, lysozyme, and lysostaphin after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate; Sugai M et al.; Bacteriolytic enzymes of different bond specificities, denatured by sodium dodecyl sulphate (SDS), were electrophoresed in polyacrylamide gels containing bacterial cells, then renatured after removal of SDS by diffusion . Enzyme activity was seen in sharp transparent bands resulting from bacteriolysis in the gels, while these sections containing bacterial cells appeared cloudy . Bacteriolytic enzymes including staphylococcal endo-beta-N-acetylglucosaminidase, lysozyme (N-acetylmuramidase), and lysostaphin (endopeptidase) were detected . The major bacteriolytic enzymes of Staphylococcus spp . were identified in gels after electrophoresis of crude enzyme preparations . This demonstrates the wide applicability of this method to the study of staphylococcal bacteriolytic enzymes . However, it should be noted that the method will fail to detect activities of bacteriolytic enzymes which are irreversibly inhibited by SDS.

Zentralbl Bakteriol, 1991 Jun, 275(2), 264 - 8
Murine CD4+ and CD8+ T cells are activated by the superantigen (SA) staphylococcal enterotoxin B (SEB) and exhibit MHC-unrestricted cytotoxicity; Miethke T et al.; Resting CD8+ and CD4+ murine T cells become efficiently activated by the superantigen (SA) staphylococcal enterotoxin B (SEB) . Limiting dilution experiments reveal that roughly every third CD8+ or CD4+ T cell could be induced to proliferate . Surprisingly, besides CD8+ T cells, also CD4+ cells acquire a high cytolytic activity upon activation with SEB . Both subpopulations only lyse MHC class II positive target cells in the presence of SEB . However the recognition of SEB by CTL is independent of the haplotype of the MHC, i.e . is MHC-unrestricted . In addition the recognition of SEB is independent of the coreceptor molecules CD4 and CD8, respectively.

Vestn Khir Im I I Grek, 1991 Jun, 146(6), 72 - 6
{Leukocyte transfusions in the treatment of sepsis}; Tulupov AN et al.; Leukotransfusion from ordinary donors and those immunized by staphylococcal anatoxin were used in complex treatment of 55 patients in a catabolic phase of different forms of sepsis . It was found that transfusion of leukosuspensions, immune ones in particular, had pronounced immunostimulating, detoxicating, antibacterial and anti-inflammatory effects . A short course of transfusions of allogenic leukocytes can be fulfilled without a selection by the HLA system.

Rev Prat, 1991 May 21, 41(15), 1363 - 7
{Primary and secondary neonatal negative coagulase staphylococcus infections}; Aujard Y et al.; As a rule, coagulase-negative staphylococcal infections in the neonate is an acquired iatrogenic infection . It usually occurs in premature infants with an in-dwelling catheter . Clinical and laboratory criteria are used to differentiate contaminated samplings from true infection . the frequency of methicillin-resistant strains justifies the use of vancomycin combined with an aminoglycoside and, in the first days, with rifampicin, all drugs administered in doses calculated for the post-conception age . This treatment is also administered in mother-to-foetus infections caused by CNS.

J Immunol Methods, 1991 May 17, 139(1), 49 - 53
Purification of staphylococcal enterotoxins A and E by immunoaffinity chromatography using a murine monoclonal antibody with dual specificity for both of these toxins; Shinagawa K et al.; An immunosorbent column was prepared by coupling a murine monoclonal antibody (MAb) with dual specificity for staphylococcal enterotoxins A (SEA) and E (SEE) to Affi-Gel 10 . Purification of both SEA and SEE from culture supernatants was carried out with the immunosorbent column using 0.2 M acetic acid containing 0.15 M NaCl as eluant . The yields obtained were approximately 76% for SEA and 70% for SEE . Purified SEA and SEE were found to be immunologically and electrophoretically homogeneous . Immunoaffinity chromatography using a MAb with dual specificity proved to be valuable in the purification of SEA and SEE, not only from the standpoint of percentage recovery, but also because of the degree of purity and the ease of purification.

Tijdschr Diergeneeskd, 1991 May 15, 116(10), 504 - 7
{Poisoning due to an ionophore anticoccidial agent in a commercial rabbit breeding farm}; Arts HT; As the percentage of rabbit feed is very small compared to the bulk of animal feeds, there is a fair chance that rabbit feed will be contaminated with constituents (additives) of batches previously prepared for other animals . Contamination of rabbit feed with monensin derived from previous runs could occur . This contamination may result in anorexia, abortion, high mortality in breeding does and outbreaks of bacterial infections (pasteurellosis and staphylococcal infection) in affected populations.

J Immunol, 1991 May 15, 146(10), 3444 - 51
IL-10 acts on the antigen-presenting cell to inhibit cytokine production by Th1 cells; Fiorentino DF et al.; Murine IL-10 (cytokine synthesis inhibitory factor) inhibits cytokine production by Th1 cell clones when they are activated under conditions requiring the presence of APC . By preincubating APC with IL-10, we demonstrate that IL-10 acts principally on APC to inhibit IFN-gamma production by Th1 clones . Moreover, IL-10 is not active when Th1 cells are stimulated with glutaraldehyde-fixed APC, which also indicates that its action involves regulation of APC function . Furthermore, IL-10 inhibits cytokine synthesis by Th1 cells stimulated with the super-antigen Staphylococcus enterotoxin B, which does not appear to require processing . Flow microfluorimetry purified splenic or peritoneal B cells and macrophages, and B cell and macrophage cell lines can present Ag to Th1 clones . However, IL-10 acts only on sorted macrophages and the macrophage cell line to suppress IFN-gamma production by Th1 clones . IL-10 does not show this effect when B cells are used as APC . In contrast, IL-10 does not impair the ability of APC to stimulate cytokine production by Th2 cells . IL-10 does not decrease IFN-gamma-induced I-Ad levels on a macrophage cell line . Inasmuch as IL-10 also inhibits IL-2-induced IFN-gamma production by Th1 cells in an Ag-free system requiring only the presence of accessory cells, these data suggest that IL-10 may inhibit macrophage accessory cell function which is independent of TCR-class II MHC interactions.

FEMS Microbiol Lett, 1991 May 15, 64(2-3), 247 - 51
Glucose control of staphylococcal enterotoxin A synthesis and location is mediated by cyclic AMP; Hallis BA et al.; The regulation of staphylococcal enterotoxin A (SEA) synthesis in a defined medium was studied using continuous culture techniques . SEA production was repressed by glucose and repression could be overcome by addition of exogenous cyclic AMP . As well as this classical catabolite repression control, addition of glucose to de-repressed steady-state cultures resulted in rapid disappearance of toxin from the medium (also mediated by loss of cyclic AMP) . When the toxin dissappeared from the medium, it was taken up again by the bacteria without apparent modification.

JAMA, 1991 May 8, 265(18), 2364 - 8
Reduced intravascular catheter infection by antibiotic bonding . A prospective, randomized, controlled trial; Kamal GD et al.; We report a prospective, randomized, controlled clinical trial to evaluate the efficacy of antibiotic-bonded catheters in reducing the incidence of intravascular catheter-related infections . Ninety-three central venous catheters and 85 arterial catheters were studied in the surgical intensive care unit . Study catheters were pretreated with the cationic surfactant tridodecylmethylammonium chloride . The anionic antibiotic, cefazolin, was bonded before insertion of the catheters by immersing them in a 50-mg/mL solution . Fourteen percent of the 81 catheters in the control group were infected, compared with 2% of the 97 antibiotic-bonded catheters . Staphylococcus epidermidis was the most common organism obtained . There was no significant difference in the number of colonized or clinically inflamed catheter insertion sites . None of the 100 antibiotic immersion solutions yielded anything on microbiologic culture . We conclude that antibiotic bonding is an efficient, safe, and cost-effective method of reducing intravascular catheter infection in patients who are in intensive care units.

Bone Marrow Transplant, 1991 May, 7(5), 405 - 9
Hemolytic uremic syndrome following bone marrow transplantation; Juckett M et al.; Bone marrow transplant (BMT) is increasingly being offered as therapy for certain hematologic and other advanced malignancies . We present one representative patient in detail and summarize data from a series of 10 patients who received a BMT and, as a late complication, developed a hemolytic uremic syndrome (HUS) . All patients presented with the triad of microangiopathic hemolytic anemia, renal insufficiency and thrombocytopenia from 30 to 875 days after BMT, and despite aggressive supportive management, plasma exchange, IgG administration and/or ex vivo staphylococcal protein A column plasma treatment, eight of 10 patients died from complications related to HUS between 11 to 139 days after diagnosis . In contrast to other reports of HUS after BMT, the present series of patients is heterogeneous with respect to underlying diagnosis, type of BMT (allogeneic or autologous), pretransplant conditioning regimen, presence of graft-versus-host disease, and use of cyclosporin . Additionally, nine of 10 patients were in clinical remission at the time of diagnosis of HUS, and six of 10 patients had achieved a complete recovery following BMT . The cause of HUS in this series of patients is probably multifactorial and related to the intensive pretransplant conditioning regimen.

DICP, 1991 May, 25(5), 478 - 84
Antimicrobial prophylaxis in coronary bypass surgery: a critical appraisal; Ariano RE et al.; The literature has been examined to assess the optimal prophylactic antimicrobial regimen for patients undergoing coronary bypass surgery . Antimicrobial surgical prophylaxis should be based on the two main potential pathogens, Staphylococcus epidermidis and S . aureus . It is unclear whether the prophylactic use of antimicrobials can or should be guided by in vitro antimicrobial susceptibility testing; data from well-performed clinical trials should be evaluated . The data fail to demonstrate consistently a significant difference within the cephalosporin class of antimicrobials with regard to prevention of infectious complications . Although it does not reach statistical difference, the trend with respect to efficacy appears to be cefuroxime, then cefamandole, and then cefazolin . The lack of significant difference among antimicrobials suggests an institution-individualized approach to the selection of the optimal antimicrobial for prophylaxis . For our facilities we recommend the following regimen: cefazolin sodium 1-2 g iv q8h for two days . There are not enough data at this time to recommend less than two days of antimicrobial prophylaxis for this type of surgery . In addition, aminoglycosides provide no added benefit when added to cephalosporins.

Ophthalmology, 1991 May, 98(5), 639 - 49; discussion 650
Role of external bacterial flora in the pathogenesis of acute postoperative endophthalmitis; Speaker MG et al.; The source of infecting bacteria is often unknown in postoperative endophthalmitis . Using techniques of molecular epidemiology, the authors demonstrate that an organism isolated from the vitreous was genetically indistinguishable from an isolate recovered from the patient's eyelid, conjunctiva, or nose in 14 (82%) of 17 cases of endophthalmitis . In two cases of Staphylococcus epidermidis endophthalmitis, after cataract surgery was performed by the same surgeon on the same day, genetic analysis demonstrated that the infecting organisms from the two patients' vitreous were distinct, and, furthermore, that in each case the external isolates were indistinguishable from the vitreous isolate . The results implicate the patients' external tissues as the source of the infecting organism in those cases of acute postoperative endophthalmitis studied . Therefore, attention should be directed toward the external tissues and their microbial flora in the prophylaxis and prevention of postoperative bacterial endophthalmitis.

Cornea, 1991 May, 10(3), 217 - 20
Contamination of donor cornea: postpenetrating keratoplasty endophthalmitis; Antonios SR et al.; Nine cases (0.41%) of bacterial and fungal endophthalmitis developed out of a total of 2,210 consecutive penetrating keratoplasties performed between November 1983 and April 1989 . Five of the nine cases (0.23%) had endophthalmitis related to donor tissue contamination . The donor tissue of these cases had a storage time of greater than 5 days . A retrospective analysis of 1,399 consecutive corneoscleral rim cultures showed a contamination rate of 29% . The most common organisms isolated were Propionibacterium 26%, diphtheroids 24%, Staphylococcus epidermidis 22%, and fungi 9% . There is a statistically significant increase (p less than 0.005) in the percentage of contaminated donor rims with a preservation time of more than five days . The risk of developing endophthalmitis is 12 times greater with a positive donor rim culture . Prolonged preservation of donor tissue can be a risk factor in developing endophthalmitis.

Eur J Immunol, 1991 May, 21(5), 1331 - 3
Evidence for T cell receptor-HLA class II molecule interaction in the response to superantigenic bacterial toxins; Fleischer B et al.; The staphylococcal enterotoxins and related microbial T cell mitogens stimulate T cells by cross-linking variable parts of the T cell receptor (TcR) with MHC class II molecules on accessory or target cells . In this report we describe that a given combination of T cell, accessory cell (AC) and toxin can be non-stimulatory . However, the same T cell can respond to the same toxin on another AC and the same AC can present the same toxin to another T cell . This indicates that in the complex formed between TcR, toxin and class II molecule an interaction between TcR and class II molecule takes place.

J Intraven Nurs, 1991 May-Jun, 14(3 Suppl P), S25 - 9
Treatment modalities for patients with HIV disease; Molaghan JB; Pharmacologic and psychosocial interventions begin at the time HIV infection is diagnosed and continue to the end of the patient's life . It is important that the nurse and patient communicate effectively with one another about the significance of being HIV positive, including disease progression and treatment modalities . Symptoms of and treatments for early infection (e.g., oral candidiasis, aphthous stomatitis, staphylococcal folliculitis, and herpes zoster) are reviewed . Pneumocystis carinii infection is a major concern; pentamidine, dapsone, and trimethoprim--sulfamethoxazole may be used for prophylaxis . Regimens and guidelines for zidovudine treatment are discussed . Promising antiretroviral therapies include ddI, ddC, CD4, protease inhibitors, and compound Q . Ultimately, vaccines may be available.

Clin Orthop, 1991 May, (266), 285 - 94
Foreign-body-associated experimental osteomyelitis induced with Bacteroides fragilis and Staphylococcus epidermidis in rabbits; Lambe DW Jr et al.; Bacteroides fragilis and Staphylococcus epidermidis, alone and in combination, were used to induce foreign-body-associated osteomyelitis in a rabbit model . In this model, a catheter, used as a foreign body, was implanted into the medullary cavity of the tibia . Only two of five animals infected with S . epidermidis alone developed culture-positive osteomyelitis, whereas all three animals infected with B . fragilis alone developed osteomyelitis . All six animals infected with both microorganisms developed culture-positive osteomyelitis . Roentgenographic and histologic evaluations confirmed the diagnosis of osteomyelitis . Transmission and scanning electron microscopy showed that when the two microorganisms are involved in a mixed infection, S . epidermidis predominates on the foreign body and B . fragilis predominates in the infected bone and marrow.

J Endod, 1991 May, 17(5), 213 - 6
Specificity of antibodies present in human periapical lesions; Kettering JD et al.; Various classes of immunoglobulins have been found in human periapical lesions . The specificity of secreted antibodies against antigens egressing from the root canal system has yet to be thoroughly investigated . The purpose of this study was to test the specificity of antibodies present in human periapical lesions . Human periapical biopsies were removed and cultured as organ culture explants . Antibodies present in the lesions were extracted in the cell culture fluids . A modified enzyme-linked immunosorbent assay was used to determine the presence, type, and concentration of different classes of antibodies against a number of commonly found bacterial species present in the root canal system . The data show the presence of specific antibodies (IgG, IgM, and IgA) against all 16 microorganisms tested . Peptostreptococcus micros, Actinomyces israelii, Staphylococcus intermedius, and Fusobacterium nucleatum produced significantly high levels of IgG antibodies in these lesions.

Vopr Pitan, 1991 May-Jun, (3), 56 - 9
{Determining the levels of staphylococcal enterotoxins types A and B in food products by the immunoenzyme method}; Fluer FS et al.; "Sandwich" variant of ELISA was used to identify staphylococcal enterotoxins (SE), types A and B, in S . aureus filtrates inducing food poisoning, in extracts of the lactic acid product for infants "Biphilin" that caused staphylococcal infection, and in foods contaminated with SE in varying concentrations . It has been shown that ELISA used for SE identification in foods permits revealing SE, types A and B, in liquid products in concentrations of 1-2 ng/ml (that is 1000-fold more sensitive, than the immunodiffusion test, 400-800-fold more sensitive than the passive hemagglutination test, and 10-fold more sensitive than the indirect passive hemagglutination test), and in solid products--in concentrations of 5-10 ng/g (after artificial contamination).

Ann Thorac Surg, 1991 May, 51(5), 827 - 8
Infected intravenous port device causing tricuspid valve regurgitation; Heinemann M et al.; Sepsis and tricuspid valve regurgitation developed in a 29-year-old man with Crohn's disease after implantation of an intravenous port device for parenteral nutrition . A thrombus, caused by Staphylococcus epidermidis, had grown along the catheter and prolapsed through the valve without affecting it . Complete removal in an open heart procedure led to quick recovery.

Nippon Saikingaku Zasshi, 1991 May, 46(3), 597 - 610
{STAPHYOGRAM, a new rapid identification kit for the aerobic, gram-positive, catalase-positive cocci--application of fluorometric microplate hybridization for the pre-identification of 386 isolates used}; Kawamura Y et al.; A new simplified test kit, STAPHYOGRAM plate, was developed for 4-hr identification of aerobic, Gram-positive and catalase-positive cocci . The plate has 18 wells, in which different dehydrated substrates and nutrients are fixed . An 18-hr agar-culture suspension of a test strain with a turbidity of McFarland No . 4 was distributed into all wells in 50-microliters quantities . After 4-hr incubation at 37C, the profile number was obtained by summarizing positive reactions . The ability of the plate to differentiate the type strains of the 30 species of the three genera in the family Micrococcaceae was confirmed . These three genera are Staphylococcus, Micrococcus and Stomatococcus . The applicability of the fluorometric microplate hybridization technique to identification of aerobic, Gram-positive and catalase-positive cocci was confirmed by homologous hybridization among the type strains of the 30 species . Thus, 386 isolates of human and animal origin were pre-identified by microplate hybridization and used for evaluating the STAPHYOGRAM plate . Of the 236 profile numbers thus obtained with the 386 isolates, 218 (92.4%) were species-proper each and all for the 15 species of Staphylococcus and Stomatococcus mucilaginosus . A total of 342 (88.6%) of the 386 isolates were given such profile numbers, and were identified without any additional test . Among the 15 species identified primarily by the results of STAPHYOGRAM plate culture, S . caprae, S . lugdunensis, S . gallinarum and S . delphini were validly published after Approved Lists of Bacterial Names . The identified strains of S . caprae (48), S . haemolyticus (46), S . capitis (35) numbered between those of S . epidermidis (67) and S . saprophyticus (31) . Profile numbers common to two species were seven (27 strains) and that to four species was one (17 strains) . These 44 strains were identified with one to three additional tests . From these results, we were convinced that the STAPHYOGRAM test plate is useful for the rapid identification of members of family Micrococcaceae . By compiling STAPHYOGRAM plate data on genetically identified strains, an exclusive list of profile numbers will soon be prepared for perfection of the kit.

Infection, 1991 May-Jun, 19(3), 127 - 30
Antibiotic usage for initial empirical treatment of infections in hospitalized patients in West Germany; Knothe H; Antibiotic usage for initial empirical treatment of infections in hospitalized patients was assessed by means of a questionnaire sent to physicians in charge of surgical and medical intensive care units, departments of neurosurgery, neurology, general surgery, thoracic surgery, internal medicine and pediatrics . Analysis of a total of 82 questionnaires filled in by the various departments revealed that the most frequently used regimens for initial empirical therapy were combinations of a broad spectrum penicillin with an amino-glycoside or of a second generation cephalosporin with an aminoglycoside in intensive care . Third generation cephalosporins ranked third among combination partners with aminoglycosides . Imipenem and fluoroquinolones were used only rarely for first line treatment . Second line treatment was most frequently with third generation cephalosporins or imipenem/cilastatin for internal wards and intensive care with an extension for staphylococcal infections with vancomycin or teicoplanin as the most frequent additional antibiotics . Patterns of antibiotic usage changed with regard to infection sites with a predominance of third generation cephalosporins or broad spectrum penicillins in combination with an aminoglycoside and metronidazole in abdominal sepsis and peritonitis . In case of pneumonia a differentiation between community acquired and hospital acquired pneumonias was made . Treatment was predominantly carried out with penicillin G, ampicillin or a second generation cephalosporin with or without the addition of an aminoglycoside in case of community acquired pneumonia . The addition of clindamycin or metronidazole was considered for suspected staphylococcal infection or aspiration pneumonia . Third generation cephalosporins were preferred for pneumonia treatment in surgical patients.

Vet Microbiol, 1991 May, 27(3-4), 263 - 75
Isolation of exfoliative toxin from Staphylococcus hyicus subsp . hyicus and its exfoliative activity in the piglet; Sato H et al.; Exfoliative toxin was isolated from the sterile cell-free filtrate of 24 h culture of Staphylococcus hyicus subsp . hyicus strain P-1 . The partial purification of exfoliative toxin produced by S . hyicus (shET) was performed by precipitation with 50-80% saturated ammonium sulfate, gel filtration on a Sephadex G-75 column and column chromatography on DEAE-cellulose . Partially purified shET (pp-shET) caused exfoliation in piglets at 8 to 12 h after intradermal or subcutaneous injection . However, heat-treated pp-shET did not cause exfoliation in piglets for up to 24 h after injection . On histopathological examination of the skin at 12 h after injection of pp-shET, an intraepidermal cleavage plane was shown between the stratum corneum and stratum granulosum and at the stratum granulosum.

Immunol Today, 1991 May, 12(5), 147 - 50
Staphylococcal-enterotoxin-dependent cell-mediated cytotoxicity; Dohlsten M et al.; T cells equipped with sophisticated TCR and MHC recognition structures, an efficient cytokine communication network and lethal cytotoxic effector functions constitute one of the bulwarks of the mammalian immune system . However, infective agents have developed strategies to undermine T-cell immunity; for example, certain bacterial toxins serve as 'superantigens' by binding to preserved determinants on MHC class-II-encoded proteins and activating T cells expressing particular sequences of TCR V beta gene products . In this paper, Mikael Dohlsten and colleagues present evidence suggesting that these bacterial superantigens direct T cells to eradicate MHC class-II-expressing antigen-presenting cells, thus counteracting specific T-cell functions.

J Neurol Neurosurg Psychiatry, 1991 May, 54(5), 454 - 6
Deficiencies in anti-acetylcholine receptor antibody measurement in myasthenia gravis; Clarke CE et al.; In a retrospective case note study of 86 patients with myasthenia gravis, 60 had an anti-acetylcholine receptor antibody assay performed by the regional immunology laboratory . Antibody was detected in 38% which compares with 66-93% in other series . Whilst the use of staphylococcal protein A to precipitate the antibody-receptor complex, rather than anti-human immunoglobulin, may be partly responsible for this low sensitivity, other methodological problems are likely to exist . It is suggested that this potentially critical assay becomes a subject for regular audit.

FEMS Microbiol Lett, 1991 May 1, 64(1), 35 - 9
Murine monoclonal antibodies reactive with staphylococcal enterotoxins A, B, C2, D, and E; Shinagawa K et al.; By fusion of mouse spleen cells immunized with five different staphylococcal enterotoxins (SEA, SEB, SEC2, SED, and SEE) with myeloma cells, we obtained 15 hybridomas producing monoclonal antibodies (mAbs) . Four mAbs were reactive with both SEA and SEE, whereas 8 mAbs were reactive with SEB and SEC2 . One mAb reacted with SEA, SED, and SEE . The other two mAbs were found to be reactive with all five serotypes of SEs . The mAbs specific for five serotypes of SEs were found to be most reactive with SED, reactive with SEA, and slightly less reactive with SEB, SEC2, and SEE . Those mAbs with specificities for all serotypes of SEs may be valuable to prepare immunoadsorbent(s) for isolation of SEs and to detect SEs in foods and clinical specimens involved in outbreaks of staphylococcal food poisoning.

Rev Esp Cardiol, 1991 May, 44(5), 306 - 12
{Active infective endocarditis complicated by paravalvular abscess . Review of 40 cases}; Gonzalez Vilchez FJ et al.; We have performed a retrospective analysis of the clinical, echocardiographic, surgical and necropsy data in 40 patients affected by infective endocarditis complicated by paravalvular abscess . The abscess developed on prosthetic valve in 13 cases, and on native valve in 27 . Aortic valve was the most affected (85%) . The clinical course was considered to be acute in 52% of the patients . Fever persisted despite of adequate antibiotic therapy in 57% . In 90%, overt heart failure were present . Staphylococcus were the most frequent causative microorganism (S . aureus, 22.5%; S . epidermidis, 20%) . Bundle branch or atrioventricular block were detected in 18 patients (sensibility, 45%; specificity, 88%) . Bidimensional echocardiography, along with Doppler techniques when necessary, detected an abscess in 81% of patients (sensibility, 80%; specificity, 84%) . Thirty patients were operated . Eight of them (26%) died, two intraoperatively . Ten patients were not operated and nine died (90%) . The difference was significant (p less than 0.001) . Only age and surgical treatment were related to early survival . Over the follow-up of the operated patients, five presented prosthetic dehiscence, but only two required a new operation 3 months after the first intervention, and none died . In conclusion, the detection of a paravalvular abscess complicating an infective endocarditis is an indication for surgical treatment . Both surgical mortality and postoperative prosthetic dehiscence are acceptable . Bidimensional echocardiography is the most reliable tool for the diagnosis of this complication.

Can J Microbiol, 1991 May, 37(5), 404 - 7
Monoclonal IgM antibody protection in mice against infection with an encapsulated strain of Staphylococcus epidermidis; Ichiman Y et al.; Passive protective activities of three different classes of monoclonal antibodies in mice against challenge with strain ATCC 31432 (capsular type I) of Staphylococcus epidermidis were examined . Monoclonal IgM antibody passively protected mice against challenge with the homologous strain, whereas monoclonal IgG1 and IgG2b antibodies did not . The protective activity of IgM was absorbed by the cell surface antigen extracted from the homologous strain but not by the antigen from heterologous strains . Rapid reduction of viable cells took place in the peritoneal cavity of mice immunized with monoclonal IgM as early as 6 h after the challenge with the homologous strain . An enzyme-linked immunosorbent inhibition assay showed there was remarkable inhibition with the homologous cell surface antigen but not with heterologous preparations from other strains . Results suggest that in the mouse the major passive protection against the S . epidermidis strain is provided by the IgM antibody to the cell surface antigen.

J Biochem (Tokyo), 1991 May, 109(5), 776 - 85
Expression of RNase Rh from Rhizopus niveus in yeast and characterization of the secreted proteins; Ohgi K et al.; The full-length cDNA encoding RNase Rh, which is secreted extracellularly by Rhizopus niveus, was isolated and its nucleotide sequence was determined . It was placed under control of the promoter of the glyceraldehyde 3-phosphate dehydrogenase gene of Saccharomyces cerevisiae in a high expression vector in yeast . Since yeast cells transformed by this plasmid poorly secreted RNase into the medium, the plasmid pYE RNAP-Rh was constructed, in which the signal sequence of RNase Rh was replaced by the prepro-sequence of aspartic proteinase-I, one of the extracellular enzymes secreted by R . niveus . Yeast cells harboring pYE RNAP-Rh produced RNase efficiently (ca . 40 micrograms/ml) into the medium . The product was a mixture of six enzymes (RNase RNAP-Rhs) having 3, 5, 9, 13, 14, and 16 additional amino acid residues attached to the amino terminus of the mature RNase Rh . The major product was the RNase with three additional amino acids at the amino terminus . Limited digestion of RNase RNAP-Rhs with staphylococcal V8 protease succeeded in shortening the various lengths of extra amino acid residues attached to the amino terminus of RNase Rh, yielding an RNase that has 3 additional amino acids at the amino terminus . It has been named RNase RNAP-Rh . The RNase RNAP-Rh showed the same specific activity and CD spectra as those of RNase Rh, suggesting that the two have similar conformations to each other around aromatic amino acid residues and the peptide backbone.

Eur J Immunol, 1991 May, 21(5), 1229 - 33
Human major histocompatibility complex class II-negative colon carcinoma cells present staphylococcal superantigens to cytotoxic T lymphocytes: evidence for a novel enterotoxin receptor; Dohlsten M et al.; The staphylococcal enterotoxins (SE) bind to major histocompatibility complex (MHC) class II molecules on target cells and activate T cells expressing particular T cell receptor V beta sequences . In this report we demonstrate that SE bind to the MHC class II- SW620, Colo320DM and WiDr human colon carcinoma cell lines and direct cytotoxic T lymphocytes (CTL) to mediate strong target cell killing . Flow cytometry analysis, immunoprecipitation and Northern blotting experiments failed to demonstrate any surface expression of HLA-DR, HLA-DP and HLA-DQ isotypes on the SW620 colon carcinoma cell line, whereas abundant expression of these isotypes was seen on Raji cells, SEB and SEC1 were efficiently presented at picomolar concentration by the MHC class II- colon carcinoma cells and MHC class II+ Raji cells, whereas SEA and SED were preferentially presented on the MHC class II+ Raji cells . An anti-HLA-DR monoclonal antibody inhibited SEB-induced CTL targeting to Raji, but did not influence the killing of SW620 cells . Our data suggests the existence of functionally active SE-binding structures on human colon carcinoma cells which are distinct from the conventional MHC class II molecules . The possibility that these putative new SE receptors play a role in the enterotoxin action of SE must be considered.

Med Clin (Barc), 1991 Apr 27, 96(16), 611 - 3
{Delayed nonspecific cutaneous hypersensitivity in extrinsic allergic alveolitis and idiopathic pulmonary fibrosis}; Orriols R et al.; BACKGROUND: A study of delayed nonspecific cutaneous hypersensitivity in extrinsic allergic alveolitis and in idiopathic pulmonary fibrosis . METHODS: 13 patients with extrinsic allergic alveolitis, 10 with idiopathic pulmonary fibrosis, 34 with sarcoidosis and a control group of 110 subjects without respiratory disease or apparent defects in cell immunity were evaluated . The skin tests of delayed hypersensitivity were carried out with five antigenic extracts (candidin, staphylococcal toxoid, tuberculin-PPD, trichophyllin, streptokinase-streptodornase) . The reactions were read after 48 hours . Delayed nonspecific cutaneous hypersensitivity was globally calculated, both quantitatively (sum of the mean diameters) and quantitatively (overall average of positive reactions) . RESULTS: No significant differences were found between the group of patients with idiopathic pulmonary fibrosis and the control group, nor between those with extrinsic allergic alveolitis and those with sarcoidosis . However, differences were found between the control group and the patients with extrinsic allergic alveolitis, both in the quantitative and the qualitative analysis . CONCLUSIONS: Delayed nonspecific cutaneous hypersensitivity in extrinsic allergic alveolitis is depressed in a similar way as in sarcoidosis, whereas it appears to be preserved in idiopathic pulmonary fibrosis . Therefore, extrinsic allergic alveolitis should be considered as another cause of reduction of delayed nonspecific cutaneous hypersensitivity as it is the case with sarcoidosis.

J Biol Chem, 1991 Apr 25, 266(12), 7339 - 44
Inhibition of recrystallization in ice by chimeric proteins containing antifreeze domains; Mueller GM et al.; Using synthetic DNA, we assembled a gene encoding a protein identical in sequence to one of the antifreeze proteins produced by the fish Pseudopleuronectes americanus (winter flounder) . To address the relationship between structure and function, we also assembled genes encoding proteins varying in sequence and length . The synthetic genes were cloned into a bacterial expression vector to generate translational fusions to the 3' end of a truncated staphylococcal protein A gene; the chimeric proteins encoded by these fusions, varying only in their antifreeze domains, were isolated from Escherichia coli . The antifreeze domains conferred the ability to inhibit ice recrystallization, which is characteristic of naturally occurring antifreeze proteins, on the chimeric proteins . The chimeric proteins varied in their effectiveness of inhibiting ice recrystallization according to the number of 11-amino acid repeats present in the antifreeze moiety . A protein with only two repeats lacked activity, while the inhibitory activity increased progressively for proteins containing three, four, and five repeats . Some activity was lost upon removal of either the salt bridge or the carboxyl-terminal arginine, but surprisingly, not when both features were absent together.

Biochemistry, 1991 Apr 16, 30(15), 3628 - 33
Deletion of the omega-loop in the active site of staphylococcal nuclease . 2 . Effects on protein structure and dynamics; Baldisseri DM et al.; It has been shown (Poole et al., 1991) that deletion of residues 44-49 from the sequence of staphylococcal nuclease (E43 SNase) results in an enzyme (E43 delta SNase) that is significantly more active than D43 SNase, an enzyme that differs from the wild-type enzyme by deletion of a single methylene group . In addition, both E43 delta SNase and D43 delta SNase are significantly more stable than their respective parent enzymes . Herein we use high-resolution 2D and 3D NMR spectroscopy to characterize the solution conformations of the four enzymes in order to better understand their differences in stability and activity . The backbone assignments of E43 SNase were extended to the three mutant proteins (uniformly 15N-enriched) by using 2D HSQC, 3D HOHAHA-HMQC, and 3D NOESY-HMQC spectra . The NOE patterns observed for E43 and D43 SNase in solution are consistent with the crystal structures of these proteins . The NOESY data further show that the intact and deleted proteins have essentially the same structures except that (a) the disordered omega-loops in the intact proteins are replaced by tight type II' turns, formed by residues 43-50-51-52, in the deleted proteins and (b) the orientation of the D43 side chain in crystalline D43 SNase differs from that found for D43 delta SNase in solution . Except for regions neighboring the omega-loops, the intact and deleted proteins show nearly identical amide 15N and 1H chemical shifts . In contrast, there are widespread, small and similar, chemical shift differences (a) between E43 SNase and D43 SNase and (b) between E43 delta SNase and D43 delta SNase.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemistry, 1991 Apr 16, 30(15), 3621 - 7
Deletion of the omega-loop in the active site of staphylococcal nuclease . 1 . Effect on catalysis and stability; Poole LB et al.; The high-resolution X-ray structure of wild-type staphylococcal nuclease (E43 SNase) suggests that Glu 43 acts a general basic catalyst to assist the attack of water on a phosphodiester substrate {Loll, P., & Lattman, E . E . (1989) Proteins: Struct., Funct., Genet . 5, 183} . Glu 43 is located at the base of the solvent-exposed and conformationally mobile omega-loop in the active site of E43 SNase having the sequence Glu43-Thr44-Lys45-His46-Pro47-Lys48- Lys49-Gly50-Val51-Glu52, where the gamma-carboxylate of Glu 52 is hydrogen bonded to the amide hydrogen of Glu 43 . With a metabolic selection for SNase activity produced in an Escherichia coli host, we detected an unexpected deletion of residues 44-49 of the omega-loop of E43 SNase in cassette mutagenesis experiments designed to randomize codons 44 and 45 in the omega-loop and increase the activity of the previously described E43D mutation (D43 SNase) . A high-resolution X-ray structure of D43 SNase has revealed that the E43D substitution significantly changes the structure of the omega-loop, reduces the interaction of the essential Ca2+ ion with its active-site ligands, and diminishes the network of hydrogen-bonded water molecules in the active site {Loll, P., & Lattman, E . E . (1990) Biochemistry 29, 6866} . This deletion of six amino acids from the omega-loop generates a protein (E43 delta SNase) having a partially solvent-exposed, surface beta-turn with the sequence Glu43-Gly50-Val51-Glu52; the structure of this beta-turn is addressed in the following article {Baldisseri et al . (1991) Biochemistry (following paper in this issue)}.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Immunol, 1991 Apr 15, 134(1), 216 - 24
Characterization of antigen-specific suppressor factors induced by staphylococcal enterotoxin B; Taub DD et al.; Previous studies from this laboratory have shown that staphylococcal enterotoxin B (SEB) has the capacity to nonspecifically induce multiple T suppressor cell populations which are capable of regulating both primary and secondary in vitro antibody responses . Additional studies have revealed that the suppressive activity of these cells is mediated, at least in part, by an I-J-restricted suppressor-inducer factor . Efforts to characterize the specificity of this inducer molecule have demonstrated the presence of multiple antigen-specific suppressor factors within SEB-stimulated supernatants . Antigen-binding molecules present within these SEB-induced factor preparations were isolated and concentrated over antigen-coupled columns . The results have demonstrated that eluates from trinitrophenyl-ovalbumin and trinitrophenyl-keyhole limpet hemocyanin-coupled columns significantly suppress both primary and secondary anti-TNP plaque-forming colony responses, whereas the filtrates from these columns demonstrated little to no inhibitory activity . The filtrates, but not eluates, from these columns exhibited significant suppressive activity for anti-poly(L-Phe,L-Glu)-poly-DL-Ala-poly-L-Lys, anti-azobenzenearsonate, and anti-sheep red blood cell antibody responses . Additional studies using eluates and filtrates from a variety of antigen-coated columns have demonstrated that suppressor factors of multiple antigenic specificities are present within SEB-induced supernatants . These studies suggest that SEB polyclonally activates antigen-specific suppressor cells which produce suppressor factors which specifically regulate the antibody response.

Cell Immunol, 1991 Apr 15, 134(1), 180 - 90
Antigen presenting ability of thymic macrophages and epithelial cells: evidence for defects in the antigen processing function of thymic epithelial cells; Ransom J et al.; We compared the antigen presenting ability of cloned thymic macrophage and epithelial cell lines using T cell hybridomas with well-characterized activation requirements . A cloned thymic epithelial cell line (3D.1), preinduced with interferon-gamma (IFN-gamma) activated the T cell hybridoma 3DO-18.3 but not the T cell hybridoma DO-11.10 . Analyses using preprocessed antigen suggest that the failure of 3D.1 to activate DO-11.10 is due to its inability to process chicken ovalbumin to produce a peptide recognized by the Ag:MHC T cell receptor of DO-11.10 . The epithelial cell line 3D.1 was able to activate DO-11.10 if the superantigen staphylococcal enterotoxin B was used for activation instead of ovalbumin . These observations indicate that IFN-gamma-induced 3D.1 expresses sufficient I-Ad molecules to activate DO-11.10 but is unable to produce the peptide of ovalbumin recognized by DO-11.10 . Furthermore, 3D.1 appears to be representative of nonmacrophage thymic stromal cells cultured in vitro, since heterogeneous cultures containing epithelial cells exhibited the same selective T cell activation characteristics . In contrast, thymic macrophage cell lines activated all T cells studied . These results suggest that there is a functional difference between the capacity of thymic epithelial cells and macrophages to process and present antigen to T cells.

Biochem J, 1991 Apr 15, 275 ( Pt 2), 399 - 405
Inhibition of luteinizing-hormone exocytosis by guanosine 5'-{gamma-thio}triphosphate reveals involvement of a GTP-binding protein distal to second-messenger generation; van der Merwe PA et al.; Dual inhibitory and stimulatory actions of guanine nucleotides on luteinizing-hormone (LH) exocytosis were observed in primary sheep gonadotropes permeabilized with staphylococcal alpha-toxin . At resting cytosolic {Ca2+}free (pCa 7), 5'-{gamma-thio}triphosphate (GTP{S}) and guanosine 5'-{beta gamma-imido}triphosphate (p{NH}ppG) stimulated rapid LH exocytosis, which was maximal between 5 and 10 min . GTP{S} and p{NH}ppG had similar potencies (50% of maximum effect at 20-50 microM), but the effect of p{NH}ppG was more prolonged . Experiments carried out in the presence of saturating concentrations of phorbol 12-myristate 13-acetate (PMA), or in PMA-desensitized cells, suggested that stimulation by p{NH}ppG is mediated by a mechanism additional to protein kinase C (PKC) activation . Furthermore, p{NH}ppG stimulated LH exocytosis in the presence of saturating cyclic AMP (cAMP) concentrations, although its effect was less than additive . However, when both PMA and cAMP were present, p{NH}ppG did not stimulate a further increase in the rate of LH exocytosis . In contrast, pretreatment of cells with GTP{S} at low {Ca2+}free markedly inhibited subsequent responses to Ca2+, cAMP, PMA, and cAMP plus PMA . This inhibitory effect required lower GTP{S} concentrations than the stimulatory effect (50% inhibition at 1-10 microM), and was not observed with p{NH}ppG . A similar inhibition was observed with adenosine 5'-{gamma-thio}triphosphate, probably by its conversion into GTP{S} . These results suggest that the stimulatory actions of guanine nucleotides can be accounted for by the combined activation of PKC and generation of cAMP, resulting from activation of conventional signal-transducing GTP-binding proteins . The inhibitory effect of GTP{S} can be clearly distinguished and indicates the involvement of a distinct GTP-binding protein in exocytosis at a site distal to second-messenger generation.

J Endod, 1991 Apr, 17(4), 143 - 6
Biosynthesis of IgG in periapical lesion explant cultures; Baumgartner JC et al.; The presence of immunoglobulins has been demonstrated in periapical inflammatory lesions associated with endodontic disease . The purpose of this study was to determine if IgG is synthesized in vitro in explant cultures of untreated periapical inflammatory lesions and to determine the level of IgG in isolated samples . Periapical lesions associated with infected root canals were removed from the roots and cultured in tissue culture medium containing tritiated amino acids . Supernatant fluids from the explant tissue cultures were passed through staphylococcal protein A affinity columns to isolate IgG . When the staphylococcal protein A eluents (24-h samples) from six periapical lesions were used in double diffusion in agarose assays, the presence of IgG was demonstrated in all the samples . Radial immunodiffusion assays to quantitate the IgG in staphylococcal protein A eluents showed that the levels of IgG detected in each successive daily supernatant fluid always decreased or else fell below the lower limits of detection . The in vitro biosynthesis of IgG in explant cultures of periapical lesions was demonstrated by the incorporation of tritiated amino acids into isolated IgG.

FEBS Lett, 1991 Apr 9, 281(1-2), 33 - 8
Complete 1H and 13C assignment of Lys and Leu sidechains of staphylococcal nuclease using HCCH-COSY and HCCH-TOCSY 3D NMR spectroscopy; Baldisseri DM et al.; Complete proton and carbon sidechain assignments are reported for 22 lysine and 11 leucine residues in staphylococcal nuclease, an enzyme with 149 residues . These assignments are readily obtained in a direct manner from the correlations observed in the 3D HCCH-COSY and HCCH-TOCSY spectra and the known protein backbone assignments . These assignments open the way to detailed studies of the sidechain structure and dynamics at the active site, in the hydrophobic core and on the surface of the protein.

Appl Environ Microbiol, 1991 Apr, 57(4), 1018 - 25
Selection of colony, plasmid, and virulence variants of Staphylococcus epidermidis NRC853 during growth in continuous cultures exposed to erythromycin; DeGuglielmo MA et al.; A continuous-culture system was developed to study changes in the structure of Staphylococcus epidermidis populations exposed to subminimum inhibitory concentrations of erythromycin . Continuous-culture experiments were carried out in a dextrose-free, tryptic soy broth medium supplemented with lactic acid and sodium lactate (MTSB-D) . The multiresistant (penicillin-, tetracycline-, and erythromycin-resistant) S . epidermidis strain NRC853 was subjected to a series of experiments: (i) growth individually in continuous culture in the absence and presence of erythromycin and (ii) growth in mixed culture with the erythromycin-susceptible S . epidermidis strain NRC852 in the absence and presence of erythromycin . Strain NRC853 produced colony morphology variants during continuous culture in the presence of 0.05 and 0.1 microgram of erythromycin per ml . Variants (A, B, and C) were different from their wild-type parent on the basis of colony size, sector pattern, and/or the ability to transmit light . A variants rapidly lost a 2.7-MDa tetracycline resistance plasmid . B and C variants formed an ermC plasmid multimer series from unit size to a 16-mer and exhibited an approximately twofold increase in erythromycin MIC over that of the wild-type parent . They slowly lost the tetracycline resistance plasmid . The small-colony B variant demonstrated an increased virulence in the neonatal mouse weight gain test and an increase in fibronectin binding compared with the wild-type parent . The presence of a competing strain drastically increased the frequency of all variants.

Surg Gynecol Obstet, 1991 Apr, 172(4), 275 - 9
Complications of indwelling central venous catheters in bone marrow transplant recipients; Moosa HH et al.; Complications associated with indwelling central venous catheters in patients undergoing bone marrow transplantation are presented . The study group was comprised of 123 patients who underwent transplantation from 1982 to 1988 in whom 139 catheters were placed . One hundred and fifteen of 139 catheters placed had a triple lumen, while 24 had a dual lumen . Percutaneous placement through the subclavian vein was used in 127 of 139 catheters with a low complication rate . Catheters remained in place for 65.0 +/- 55.5 days (mean +/- standard deviation) . One hundred and eight catheters remained functional during the course of treatment for these patients and lasted until elective removal or until the patient died . Thirty-one catheters were removed because of a complication . Infection was the most common complication, occurring in 22 catheters . Seven infected catheters were salvaged with antibiotic therapy . Coagulase negative staphylococcus was the most frequent organism identified . Mechanical problems causing catheter malfunction, migration, thrombosis and cuff erosion occurred in 19 catheters; 15 were removed . We concluded that double and triple lumen Silastic (silicone rubber) catheters provide relatively safe and effective access for bone marrow transplant recipients . The triple lumen catheter provides an additional venous access port that facilitates intensive care . The duration of catheter function is sufficient in most patients for therapy during and after bone marrow transplantation . Infection is the most common complication; our results indicate that catheters can be maintained and salvaged in some patients with septicemia . Noninfectious complications are less frequent, although catheters with persistent mechanical problems are less likely to be salvaged.

J Immunol, 1991 Apr 1, 146(7), 2310 - 5
Transcriptional activation of IL-1 beta and tumor necrosis factor-alpha genes by MHC class II ligands; Trede NS et al.; Ligands that bind MHC class II (Ia) molecules, including staphylococcal exotoxins (SE) and mAb induce IL-1 and TNF secretion in human monocytes and monocytic cell lines . In this study, we have analyzed the mechanisms by which SE induce IL-1 beta and TNF-alpha production . Treatment of human peripheral blood monocytes with staphylococcal exotoxin B and toxic shock syndrome toxin-1 resulted in a biphasic increase of IL-1 beta mRNA that lasted more than 12 h and in a more transient rise in TNF-alpha mRNA . A F(ab) preparation of the anti-HLA DR mAb L243 also caused a significant increase in monokine mRNA levels . Stimulation of a monocytic cell line, THP-1, with staphylococcal exotoxin B and toxic shock syndrome toxin-1 induced a rapid rise in IL-1 beta and TNF-alpha mRNA levels . This response peaked at 1 to 3 h poststimulation and remained detectable at 12 h . Nuclear run-on transcription assays demonstrated that SE cause transcriptional activation of the IL-1 beta and TNF-alpha genes . This transcriptional activation did not require de novo protein synthesis as it was not inhibited by the protein synthesis inhibitor cycloheximide . These results define an important function for Ia molecules as regulators of cytokine gene expression and add to the understanding of the changes in cellular function induced by SE.

J Neurosurg, 1991 Apr, 74(4), 656 - 9
Shunt nephritis . Case report; Rifkinson-Mann S et al.; The incidence of shunt nephritis has decreased over the past several years due to the increased use of ventriculoperitoneal (VP) shunts rather than the ventriculovascular systems for which this complication was originally reported in 1965 . Despite this trend, the syndrome has been reported in cases of VP shunting and, for this reason, merit a renewed look . Shunt nephritis is thought to be secondary to immune complex formation and deposition in the kidney in response to Staphylococcus epidermidis . The diagnostic workup and management of this disease is discussed.

Cell Immunol, 1991 Apr 1, 133(2), 446 - 55
Activation of murine T cells by staphylococcal enterotoxin E: requirement of MHC class II molecules expressed on accessory cells and identification of V beta sequence of T cell receptors in T cells reactive to the toxin; Uchiyama T et al.; We investigated a mechanism leading to activation of murine T cells by staphylococcal enterotoxin E (SEE) . L cells transfected with I-Ab genes but not control L cells supported IL-2 production by SEE-induced C57BL/6 T lymphoblasts upon restimulation with SEE . mAb to I-Ab markedly inhibited the above response . Flow cytometric analyses showed that SEE-induced C57BL/6 T lymphoblasts are composed of both CD4+ T cells and CD8+ T cells, and that larger parts of them bore V beta 11 (40-75%) . mAb to V beta 11 markedly inhibited the SEE-induced proliferative response and IL-2 production by T cells . Analysis of SEE-induced IL-2 production in spleen cells from various mouse strains showed that C57BL/6 and B10.A(4R) mice (I-E, not expressed; V beta 11+ T cells, normally generated) are highly responsive to SEE . In contrast, BALB/c, C3H/HeN, (C57BL/6 x BALB/c or C3H/HeN) F1 mice (I-E, normally expressed and V beta 11+ T cells, deleted), and SJL and C57L mice (V beta 11 genes, deleted) are weakly responsive to SEE . The results indicate that SEE activates mainly T cells bearing V beta 11 in physical association with MHC class II molecules expressed on AC . In addition, the results indicate that SEE activates both CD4+ T cells and CD8+ T cells.

J Clin Microbiol, 1991 Apr, 29(4), 738 - 44
Identification of Staphylococcus species and subspecies with the MicroScan Pos ID and Rapid Pos ID panel systems; Kloos WE et al.; The accuracies of the MicroScan Pos ID and Rapid Pos ID panel systems (Baxter Diagnostic Inc., MicroScan Division, West Sacramento, Calif.) were compared with each other and with the accuracies of conventional methods for the identification of 25 Staphylococcus species and 4 subspecies . Conventional methods included those used in the original descriptions of species and subspecies and DNA-DNA hybridization . The Pos ID panel uses a battery of 18 tests, and the Rapid Pos ID panel uses a battery of 42 tests for the identification of Staphylococcus species . The Pos ID panel has modified conventional and chromogenic tests that can be read after 15 to 48 h of incubation; the Rapid Pos ID panel has tests that use fluorogenic substrates or fluorometric indicators, and test results can be read after 2 h of incubation in the autoSCAN-W/A . Results indicated that both MicroScan systems had a high degree of congruence (greater than or equal to 90%) with conventional methods for the species S . capitis, S . aureus, S . auricularis, S . saprophyticus, S . cohnii, S . arlettae, S . carnosus, S . lentus, and S . sciuri and, in particular, the subspecies S . capitis subsp . capitis and S . cohnii subsp . cohnii . The Rapid Pos ID panel system also had greater than or equal to 90% congruence with conventional methods for S . epidermidis, S . caprae, S . warneri subsp . 2, S . xylosus, S . kloosii, and S . caseolyticus . For both MicroScan systems, congruence with conventional methods was 80 to 90% for S . haemolyticus subsp . 1, S . equorum, S . intermedius, and S . hyicus; and in addition, with the Rapid Pos ID panel system congruence was 80 to 89% for S . capitis subsp . ureolyticus, S . warneri subsp . 1, S . hominis, S . cohnii subsp . urealyticum, and S . simulans . The MicroScan systems identified a lower percentage (50 to 75%) of strains of S . lugdunensis, S . gallinarum, S . schleiferi, and S . chromogenes, although the addition of specific tests to the systems might increase the accuracy of identification significantly.

J Chemother, 1991 Apr, 3(2), 79 - 82
Transient state kinetic studies on catalysis and inhibition of staphylococcal beta-lactamase; Virden R; The interaction of beta-lactamase with a beta-lactam substrate may, in addition to hydrolysis of the beta-lactam by way of an acylenzyme intermediate, lead to transient or permanent inhibition through one or more branches in the catalytic pathway . The balance between these processes depends on the interplay between the chemical reactivities of interacting groups and on effects on protein conformation and stability but the molecular basis is not fully understood . Examples of physical and kinetic probes of some of the mechanisms are discussed.

Anal Biochem, 1991 Apr, 194(1), 110 - 20
Functionalized membrane supports for covalent protein microsequence analysis; Coull JM et al.; Methods were developed for high yield covalent attachment of peptides and proteins to isothiocyanate and arylamine-derivatized poly(vinylidene difluoride) membranes for solid-phase sequence analysis . Solutions of protein or peptide were dried onto 8-mm membrane disks such that the functional groups on the surface and the polypeptide were brought into close proximity . In the case of the isothiocyanate membrane, reaction between polypeptide amino groups and the surface isothiocyanate moieties was promoted by application of aqueous N-methylmorpholine . Attachment of proteins and peptides to the arylamine surface was achieved by application of water-soluble carbodiimide in a pH 5.0 buffer . Edman degradation of covalently bound polypeptides was accomplished with initial and repetitive sequence yields ranging from 33 to 75% and 88.5 to 98.5%, respectively . The yields were independent of the sample load (20 pmol to greater than 1 nmol) for either surface . Significant loss of material was not observed when attachment residues were encountered during sequence runs . Application of bovine beta-lactoglobulin A chain, staphylococcus protein A, or the peptide melittin to the isothiocyanate membrane allowed for extended N-terminal sequence identification (35 residues from 20 pmol of beta-lactoglobulin) . A number of synthetic and naturally occurring peptides were sequenced to the C-terminal residue following attachment to the arylamine surface . In one example, 10 micrograms of bovine alpha-casein was digested with staphylococcal protease V8 and the peptides were separated by reverse-phase chromatography . Peptide fractions were then directly applied to arylamine membrane disks for covalent sequence analysis . From as little as 2 pmol of initial signal it was possible to determine substantial sequence information (greater than 10 residues).

Int J Syst Bacteriol, 1991 Apr, 41(2), 284 - 9
Staphylococcus cohnii subspecies: Staphylococcus cohnii subsp . cohnii subsp . nov . and Staphylococcus cohnii subsp . urealyticum subsp . nov; Kloos WE et al.; Two major subspecies of Staphylococcus cohnii, namely S . cohnii subsp . cohnii, from humans, and S . cohnii subsp . urealyticum, from humans and other primates, are described on the basis of a study of 14 to 25 strains and 18 to 33 strains, respectively . DNA-DNA hybridization studies conducted in our laboratory in 1983 (W . E . Kloos and J . F . Wolfshohl, Curr . Microbiol . 8:115-121, 1983) demonstrated that strains representing the different subspecies were significantly divergent . S . cohnii subsp . urealyticum can be distinguished from S . cohnii subsp . cohnii on the basis of its greater colony size; pigmentation; positive urease, beta-glucuronidase, and beta-galactosidase activities; delayed alkaline phosphatase activity; ability to produce acid aerobically from alpha-lactose; and fatty acid profile . The type strain of S . cohnii subsp . cohnii is ATCC 29974, the designated type strain of S . cohnii Schleifer and Kloos 1975b, 55 . The type strain of S . cohnii subsp . urealyticum is ATCC 49330.

J Antimicrob Chemother, 1991 Apr, 27 Suppl B, 61 - 7
Prophylaxis, cost and effectiveness of therapy of infections caused by gram-positive organisms in neutropenic children; Schaison GS et al.; Gram-positive infections are being reported with increasing frequency in children with haematological malignancies . Staphylococcus epidermidis, once considered a non-pathogenic skin contaminant, is emerging as a major cause of severe infection . However, in infants Gram-negative septicaemias are more frequent than in older children . A teicoplanin and ceftriaxone combination was assessed for use as empirical therapy of febrile episodes in neutropenic children with acute leukaemias . Of 47 patients, fever was of unknown origin in 21, and documented in 26 with 28 strains isolated; 19 Gram-positive (all sensitive to teicoplanin) and nine Gram-negative . Within 48 h, 41 patients became afebrile and the pathogen was cleared if initially present . Mean duration of treatment was 16 days . Febrile relapse occurred in 24 patients with eight documented superinfections . The need for prophylactic cover against Gram-positive organisms at the time of intravenous catheter insertion is questionable . We studied 71 patients who were randomly allocated to receive teicoplanin when the central line was inserted and if febrile, with added ceftriaxone and amikacin (arm A) or the tri-antibiotic regimen when fever occurred (arm B) . In arm A a febrile episode occurred after ten days in 34/35 patients with only one Gram-positive organism isolated . In arm B a febrile episode occurred in all 36 patients after five days and ten Gram-positive strains were isolated . Those patients in arm A also received fluconazole . Amphotericin B was administered in cases of failure or relapses in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Mikrobiyol Bul, 1991 Apr, 25(2), 138 - 43
{Staphylococcal protein A, bacteriocin, DNase and lactamases of methicillin-resistant Staphylococcus}; Sultan N et al.; 231 Methicillin-Resistant Staphylococcus (MRS) and 76 Methicillin-Susceptible Staphylococcus (MSS) strains were investigated for Staphylococcal Protein A, bacteriocin, DNase and beta lactamase production properties . It was found that 73.6% of the strains of MRS were positive for protein A, 3.8% for bacteriocin, 76.2% for DNase and 84.4% for beta lactamase production . And it was found that 61.8% of MSS strains were positive for protein A, 11.8% for bacteriocin, 55.2% for DNase and 57.9% for beta lactamase production . Staphylococcal protein A, DNase and lactamase production . Staphylococcal protein A, DNase and lactamase production were found to be significantly higher in MRS strains than MSS strains and bacteriocin production was found to be higher in MSS strains than MRS strains according to Chi Square Test.

J Vet Med Sci, 1991 Apr, 53(2), 223 - 7
Immunological cross-reactivity of the fragments of staphylococcal enterotoxins A and E generated by digestion of proteolytic enzymes; Shinagawa K et al.; Three major fragments were generated by limited digestion of staphylococcal enterotoxins A (SEA) and E (SEE) with papain, whereas five major fragments were generated by limited digestion with staphylococcal protease V8 (V8) . All of these fragments were detected by immunoblotting with polyclonal anti-SEA and -SEE sera . Some of generated fragments were detected by monoclonal antibodies (MAbs) with specificities for SEA (A-111 and A-211), SEE (E-142), or both (AE-32, AE-37, and AE-53) . This indicates that fragments of SEA and SEE containing the type-specific and cross-reacting epitopes may be generated by digestion of the toxins with either papain or V8.

J Gen Microbiol, 1991 Apr, 137 ( Pt 4), 977 - 81
Cloning and sequence analysis of a plasmid-encoded chloramphenicol acetyltransferase gene from Staphylococcus intermedius; Schwarz S et al.; The chloramphenicol acetyltransferase gene (cat) of a 3.9 kb chloramphenicol resistance (CmR) plasmid from Staphylococcus intermedius, designated pSCS1, was cloned into an Escherichia coli plasmid vector . Sequence analysis revealed a high degree of base similarity with the cat gene of the S . aureus CmR plasmid pC221 but there were several differences in the regulatory region . A lesser degree of similarity was observed between the cat gene of the S . intermedius plasmid and the cat gene of the S . aureus plasmid pC194.

Appl Microbiol Biotechnol, 1991 Apr, 35(1), 10 - 3
Studies on the production of lipase from recombinant Staphylococcus carnosus; Falk MP et al.; Production of lipase from recombinant Staphylococcus carnosus pLipPS1 was studied in standard stirred tank bioreactors . Only low lipase activity was obtained under conventional operating conditions, i.e., moderate to high stirring speeds and aeration rates for keeping the dissolved oxygen concentration at high levels . Additional targetted experiments indicated that the reason for the observed low lipase activity is lipase inactivation due to surface forces and shear stress at the gas/liquid interface . Therefore, a cultivation strategy is proposed that minimizes gas/liquid interfacial area and maximizes the driving concentration for O2 mass transfer by controlling the dissolved oxygen to low values by gentle stirring and low aeration rates . Thus, high lipase activities can be obtained even in larger scale standard stirred tank bioreactors.

Biotechnology (N Y), 1991 Apr, 9(4), 363 - 6
Facilitated in vitro refolding of human recombinant insulin-like growth factor I using a solubilizing fusion partner; Samuelsson E et al.; We describe a new approach to refolding recombinant proteins in which an affinity fusion partner, consisting of two IgG-binding domains (ZZ) derived from staphylococcal protein A, is used to solubilize misfolded molecules before, during and after reduction and reoxidation . We show that human insulin-like growth factor I (IGF-I) can be refolded as a fusion protein at a concentration as high as 1-2 mg/ml without the use of denaturing agents . A process scheme suitable for large scale application is described in which the yield of correctly folded human IGF-I with full biological activity is substantially increased.

Minerva Chir, 1991 Mar 31, 46(6), 269 - 72
{Importance of Staphylococcus epidermidis in the bacterial colonization of abdominal drains in surgical patients}; Chisena S et al.; Bacterial adherence to biomaterials as an element of clinical relevance is a well-known factor in the pathogenesis of sepsis . Drainages, intravascular catheters, surgical prostheses and other devices are susceptible to bacterial colonization with clinical consequences . In the last few years attention has been paid to coagulase-negative Staphylococcus (S . epidermidis), mainly to some strains able to produce a highly adhesive polysaccharide substance, called glycocalix or slime . This promotes adherence either interbacterial either between bacteria and biologic tissues or synthetic materials acting as a pathogenetic factor in sepsis being able to increase bacterial resistance to phagocytes and antibiotics . Bacterial contamination of 40 abdominal drainage tubes in patients operated in elective and in emergency surgery for various pathology has been evaluated . Sonication of the tip of the drainage was utilized in order to promote the detachment of adherent colonies and its effectiveness was compared to that of microcentrifugation . Culture of 25 drainages (62.5%) showed no bacterial contamination; 7 drainages (17.5%) have proved to be colonized by S . epidermidis, in 4 cases the isolated strains were also methicillin-resistant, 2 of which slime-producing . Out of the 7 drainages colonized by S . epidermidis, 4 were removed from patients operated in emergency: none of the isolated strains was slime-producing . Six drainages (15%) were colonized by Gram+ bacteria (S . fecalis, P-sensible cocci, rods), 1 (2.5%) by E . coli and 1 (2.5%) by P . aeruginosa, S . epidermidis appears to be the chief contaminant of abdominal drainages, especially in emergency surgery; slime production has always been observed in methicillin-resistant strains: this confirms the hypothesis that slime production is typical of specialized, virulent strains.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1991 Mar 25, 266(9), 5827 - 33
The conformation of SecA, as revealed by its protease sensitivity, is altered upon interaction with ATP, presecretory proteins, everted membrane vesicles, and phospholipids; Shinkai A et al.; Interactions between SecA and cellular components involved in the translocation of secretory proteins across the cytoplasmic membrane of Escherichia coli were studied by examining changes in the sensitivity of SecA to staphylococcal protease V8 . In the presence of ATP, the amino-terminal 95-kDa portion of the SecA molecule became highly resistant to V8 digestion . Adenosine 5'-(gamma-thio)triphosphate (ATP gamma S) and ADP were as effective as ATP . For the effect, ATP could be partly replaced by CTP and UTP, but not GTP, as in the case of the protein translocation reaction . In the presence of proOmpA, a presecretory protein, on the other hand, SecA became more sensitive to V8 digestion . The signal peptide region was involved in this effect . The V8-digestion profile in the presence of both proOmpA and ATP or ADP was the same as that in the presence of proOmpA alone . Consistently, proOmpA-induced discharge of ADP or ATP gamma S from SecA was observed by means of flow dialysis . SecA-deprived everted membrane vesicles and an E . coli phospholipid mixture were also effective in making SecA more sensitive to V8 digestion . Among the phospholipids, phosphatidylglycerol and cardiolipin were effective, whereas phosphatidylethanolamine was not . It is suggested that SecA directly interacts with these cellular components and the interactions result in changes in the conformation of SecA . The physiological significance of such interactions in protein secretion is discussed.

N Engl J Med, 1991 Mar 21, 324(12), 795 - 800
Improvement in the diagnosis of abscesses associated with endocarditis by transesophageal echocardiography; Daniel WG et al.; BACKGROUND . Echocardiography is recognized as the method of choice for the noninvasive detection of valvular vegetations in patients with infective endocarditis, with transesophageal echocardiography being more accurate than transthoracic echocardiography . The diagnosis of associated abscesses by transthoracic echocardiography is difficult or even impossible in many cases, however, and it is not known whether transesophageal echocardiography is any better . METHODS . To determine the value of transesophageal echocardiography in the detection of abscesses associated with endocarditis, we studied prospectively by two-dimensional transthoracic and transesophageal echocardiography 118 consecutive patients with infective endocarditis of 137 native or prosthetic valves that was documented during surgery or at autopsy . RESULTS . During surgery or at autopsy, 44 patients (37.3 percent) had a total of 46 definite regions of abscess . Abscesses were more frequent in aortic-valve endocarditis than in infections of other valves, and the infecting organism was more often staphylococcus (52.3 percent of cases) in patients with abscesses than in those without abscesses (16.2 percent) . The hospital mortality rate was 22.7 percent in patients with abscesses, as compared with 13.5 percent in patients without abscesses . Whereas transthoracic echocardiography identified only 13 of the 46 areas of abscess, the transesophageal approach allowed the detection of 40 regions (P less than 0.001) . Sensitivity and specificity for the detection of abscesses associated with endocarditis were 28.3 and 98.6 percent, respectively, for transthoracic echocardiography and 87.0 and 94.6 percent for transesophageal echocardiography; positive and negative predictive values were 92.9 and 68.9 percent, respectively, for the transthoracic approach and 90.9 and 92.1 percent for the transesophageal approach . Variation between observers was 3.4 percent for transthoracic and 4.2 percent for transesophageal echocardiography . CONCLUSIONS . The data indicate that transesophageal echocardiography leads to a significant improvement in the diagnosis of abscesses associated with endocarditis . The technique facilitates the identification of patients with endocarditis who have an increased risk of death and permits earlier treatment.

Biochemistry, 1991 Mar 19, 30(11), 2876 - 82
Exchange of phosphoryl groups between HPr molecules of the phosphoenolpyruvate-dependent phosphotransferase system is an autocatalytic process; van Dijk AA et al.; HPr, a central component of the phosphoenolpyruvate-dependent phosphotransferase system, can exist in Escherichia coli in a phosphorylated (PHPr) and a nonphosphorylated form . We show that, beside the normal transfer of the phosphoryl group from PHPr to enzymes II and III, PHPr can phosphorylate other HPr molecules in an autocatalytic exchange reaction . The reaction is very fast but is inhibited by labeling the protein with Bolton-Hunter reagent . We demonstrate that the exchange reaction can be used to determine the delta G degree of the phosphoryl group of mutant forms of PHPr relative to wild-type PHPr . Two HPr mutants were constructed by site-directed mutagenesis, HPr P11E and HPr E68A . Both show altered phosphoryl group potentials but show no significantly altered KM or Vmax values compared to wild-type HPr, illustrating the sensitivity of the exchange process . The exchange reaction does not occur between HPr from E . coli and HPr from Staphylococcus carnosus.

J Biol Chem, 1991 Mar 15, 266(8), 4719 - 25
Peptidoglycan and lipopolysaccharide bind to the same binding site on lymphocytes; Dziarski R; Bacterial cell wall peptidoglycan (PGN) and lipopolysaccharide (LPS), which are both macrophage activators and polyclonal B cell mitogens, were shown to bind to the same dominant 70-kDa 6.5 pI protein on the surface of mouse B lymphocytes . This conclusion was supported by the following results: (a) the PGN- and LPS-binding proteins co-migrated following photoaffinity cross-linking and two-dimensional polyacrylamide gel electrophoresis; (b) cross-linking of PGN to this 70-kDa protein was competitively inhibited by LPS (IC50 = 7.3 microM), LPS from a deep rough mutant (IC50 = 6.9 microM), and lipid A (IC50 = 18-72 microM); (c) cross-linking of LPS to this 70-kDa protein was competitively inhibited by polymeric soluble PGN (IC50 = 0.09 microM) and sonicated high Mr PGN (IC50 = 0.6 microM); (d) cross-linking of both PGN and LPS to this 70-kDa protein was also competitively inhibited by dextran sulfate (IC50 = 115-124 microM); (e) cross-linking of both PGN and LPS to this 70-kDa protein was inhibited by a (GlcNAc)2-specific lectin; and (f) peptide maps of the 70-kDa proteins digested with chymotrypsin, subtilisin, staphylococcal protease V, or papain were identical for PGN- and LPS-binding proteins and unique for each enzyme . Based on competitive inhibition experiments, binding of PGN to the 70-kDa protein was 20-1200 times stronger than the binding of LPS or lipid A on a per mol basis . However, when aggregated micellar structures of LPS or lipid A were considered, the avidities of LPS and PGN binding were similar . These results demonstrate binding of PGN and LPS to the same 70-kDa protein on lymphocytes and suggest that the binding is specific for the (GlcNAc-MurNAc)n backbone of PGN and the (GlcNAc)2 part of lipid A.

Cancer, 1991 Mar 15, 67(6), 1570 - 5
Association of interleukin-2 therapy with staphylococcal bacteremia; Richards JM et al.; The authors prospectively monitored patients undergoing leukapheresis for peripheral stem cell harvesting (PSCH) or lymphokine activated killer (LAK) cell generation for 3 weeks after catheter placement for evidence of local or systemic infections . Over a 1-year period, 16 patients underwent leukapheresis for PSCH in preparation for autologous bone marrow transplantation (ABMT) . The original catheters remained in place an average of 20 days without any documented infections . Seventeen patients underwent leukapheresis as part of a low-dose interleukin-2 (IL-2) treatment for LAK cell generation, and their catheters remained in place an average of 20.2 days with three documented episodes of bacteremia (18%) . Eight patients treated with high-dose IL-2 also underwent leukapheresis for LAK cell generation and their catheters remained in place an average of 12 days with three documented episodes of bacteremia (38%) . In all cases of bacteremia, Staphylococcus species were isolated from the blood . The IL-2 exposure level was associated with the risk of bacteremia (P = 0.01) . Other potential risk factors (e.g., number of pheresis procedures, complement level, serum immunoglobulin levels, absolute neutrophil count) were not related to this risk.

Gene, 1991 Mar 15, 99(2), 243 - 8
Synthesis and secretion of a fibrinolytically active tissue-type plasminogen activator variant in Escherichia coli; Waldenstrom M et al.; A gene encoding a variant (lacking amino acids 6-173) of human tissue-type plasminogen activator (t-PA), consisting only of the second kringle domain (K2) and the serine protease domain (P), was fused to a DNA segment coding for the signal peptide of staphylococcal protein A and a synthetic gene coding for a protein with ability to bind immunoglobulin G (IgG) . The fusion protein which was synthesized in Escherichia coli and secreted into the growth medium, was found to be fibrinolytically active . Purification of the fusion protein was performed in a single step by affinity chromatography with immobilized IgG . Enzymatically active K2P was liberated from the fusion protein by cleavage at a unique Asn-Gly dipeptide sequence using hydroxylamine . These results demonstrate that a variant of human t-PA can be synthesized and secreted by E . coli as a fibrinolytically active fusion protein, which upon specific cleavage yields an active variant t-PA of the expected size.

Biochemistry, 1991 Mar 12, 30(10), 2698 - 706
Folding of staphylococcal nuclease A studied by equilibrium and kinetic circular dichroism spectra; Sugawara T et al.; The urea-induced unfolding of staphylococcal nuclease A has been studied by circular dichroism both at equilibrium and by the kinetics of unfolding and refolding (pH 7.0 and 4.5 degrees C), as a function of Ca2+ and thymidine 3',5'-diphosphate (pdTp) concentration . The results are as follows . (1) The unfolding transition is shifted to higher concentrations of urea by Ca2+ and pdTp, and the presence of both ligands further stabilizes the protein . (2) In the first stage of kinetic refolding, the peptide ellipticity changes rapidly within the dead time of stopped-flow measurement (15 ms), indicating accumulation of a transient intermediate . This intermediate is remarkably less stable than those of other globular proteins previously studied . (3) Dependence of the folding and unfolding rate constants on urea concentration indicates that the critical activated state of folding ("transition state") has considerable structural organization . The transition state does not, however, have the capacity to bind Ca2+ and pdTp, as indicated by the effects of these ligands on the unfolding rate constant . (4) There are at least four different phases in the refolding kinetics in native conditions below 1 M urea . In the absence of pdTp, there are two phases in unfolding, while in the presence of pdTp the unfolding kinetics show a single phase . Some characteristics of the transient intermediate and of the transition state for folding are discussed.

Acta Chir Orthop Traumatol Cech, 1991 Mar, 58(1-2), 37 - 46
{Use of fluoroquinolone chemotherapy in orthopedics and traumatology}; Krcmery V Jr; In the submitted review the authors analyze hitherto published results with the use of new antimicrobial chemotherapeutic drugs--fluoroquinolones in the treatment of infections of the bones, joints, soft tissues and also in the peroperative prophylaxis in orthopaedics and traumatology . Quinolones are one of the drugs of choice which may be successful in the treatment of staphylococcal and gram-negative chronic osteomyelitis which are resistant to classical treatment and also the drug of choice in the treatment of septic arthritis and ostitisPublication Types:

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