Staphylococcus

J Bacteriol, 1995 May, 177(10), 2827 - 33
Molecular characterization of the staphylococcal multidrug resistance export protein QacC; Paulsen IT et al.; The QacC polypeptide is a member of a family of small membrane proteins which confer resistance to toxic compounds . The staphylococcal qacC gene confers resistance to toxic organic cations via proton-dependent export . The membrane topology of the QacC polypeptide was investigated by constructing and analyzing a series of qacC-phoA and qacC-lacZ fusions . From these analyses, most of the predicted features of the QacC protein were verified, although data regarding the possible orientation of the COOH region were not conclusive . The role of the sole cysteine residue, Cys-42, in QacC was studied by using the sulfhydryl reagent N-ethylmaleimide and site-directed mutagenesis . N-Ethylmaleimide was shown to inhibit qacC-mediated ethidium export . Multiple amino acid substitutions were made for Cys-42, and mutations at this location had various effects on resistance specificity . This suggests that the Cys-42 residue may be located near a region of QacC that is involved in substrate recognition . Mutagenesis of conserved residues in QacC indicated that Tyr-59 and Trp-62 also play an essential structural or functional role in QacC.

Arch Dermatol, 1995 May, 131(5), 552 - 5
Cultures of skin biopsy tissue from immunocompromised patients with cancer and rashes; Chren MM et al.; BACKGROUND AND DESIGN: Microbiological cultures of skin biopsy tissue are often recommended in immunocompromised patients with cancer and rashes, but in a previous study, they were usually sterile or grew clinically insignificant organisms . To examine the use and bacteriological results of these cultures more comprehensively, we reviewed records from all immunocompromised adults with cancer and acute rash on which skin biopsy was performed during 39 months on a bone marrow transplantation/acute leukemia unit of a university hospital (108 episodes of rash in 80 patients) . RESULTS: Of the 158 cultures that were performed, one (1%; 95% confidence interval {CI}, 0% to 4%) was a true positive; 11 (7%; 95% CI, 3% to 13%) were false positive; 143 (91%; 95% CI, 87% to 95%) were true negative; and three (2%; 95% CI, 1% to 6%) were false negative . Thus, the sensitivity of culture was 0.25, and the specificity was 0.93 . Coagulase-negative Staphylococcus was the single pathogenic organism that grew, yet was judged to be a contaminant in three episodes . Among the 95 rashes in which fewer than four types of culture were performed, viral culture may have been helpful in one case (1%; 95% CI, 0% to 6%) . CONCLUSIONS: Cultures of skin tissue from immunocompromised cancer patients with rashes were often unable to diagnose infection or the absence of infection . Clinical judgement was crucial to the interpretation of culture results.

J Bacteriol, 1995 May, 177(9), 2408 - 15
Characterization of a genetic locus essential for maltose-maltotriose utilization in Staphylococcus xylosus; Egeter O et al.; A genetic locus from Staphylococcus xylosus involved in maltose-maltotriose utilization has been characterized . The chromosomal region was identified by screening a genomic library of S . xylosus in Escherichia coli for sucrose hydrolase activity . Nucleotide sequence analysis yielded two open reading frames (malR and malA) encoding proteins of 37.7 and 62.5 kDa, respectively . MalR was found to be homologous to the LacI-GalR family of transcriptional regulators, and MalA showed high similarity to yeast alpha-1,4-glucosidases and bacterial alpha-1,6-glucosidases . Inactivation of malA in the genome of S . xylosus led to a maltose-maltotriose-negative phenotype . In cell extracts of the mutant, virtually no glucose release from maltose and short maltodextrins was detectable . Inactivation of malA in a sucrose-6-phosphate hydrolase-deficient S . xylosus strain resulted in the complete loss of the residual sucrose hydrolase activity . The MalA enzyme has a clear preference for maltose but is also able to release glucose from short maltosaccharides . It cannot cleave isomaltose . Therefore, malA encodes an alpha-1,4-glucosidase or maltase, which also liberates glucose from sucrose . Subcloning experiments indicated that malA does not possess its own promoter and is cotranscribed with malR . Its expression could not be stimulated when maltose was added to the growth medium . Chromosomal inactivation of malR led to reduced maltose utilization, although alpha-glucosidase activity in the malR mutant was slightly higher than in the wild type . In the mutant strain, maltose uptake was reduced and inducibility of the transport activity was partially lost . It seems that MalR participates in the regulation of the gene(s) for maltose transport and is needed for their full expression . Thus, the malRA genes constitute an essential genetic locus for maltosaccharide utilization in S . xylosus

J Immunol, 1995 May 1, 154(9), 4883 - 94
Increased frequency of gamma delta T cells in cerebrospinal fluid and peripheral blood of patients with multiple sclerosis . Reactivity, cytotoxicity, and T cell receptor V gene rearrangements; Stinissen P et al.; Infiltrating gamma delta T cells are potentially involved in the central nervous system demyelination in multiple sclerosis (MS) . To further study this hypothesis, we analyzed the frequency and functional properties of gamma delta T cells in peripheral blood (PB) and paired cerebrospinal fluid (CSF) of patients with MS and control subjects, including patients with other neurologic diseases (OND) and healthy individuals . The frequency analysis was performed under limiting dilution condition using rIL-2 and PHA . After PHA stimulation, a significantly increased frequency of gamma delta T cells was observed in PB (14.7 x 10(-4)) and in CSF (15.8 x 10(-4)) of MS patients as compared with 4.3 x 10(-4) in PB and 3.9 x 10(-4) detected in CSF of patients with OND . The frequency was represented equally in OND patients and normal individuals . Similarly, the IL-2-responsive gamma delta T cells occurred at a higher frequency in PB of control subjects (1.1 x 10(-4)) in OND patients and 1.5 x 10(-4) in normal individuals) . Forty-three percent (13 of 30) of the gamma delta T cell clones isolated from PB and CSF of MS patients responded to heat shock protein (HSP70) but not HSP65, whereas only 2 of 30 control gamma delta T cell clones reacted to the HSP . The majority of the gamma delta T cell clones were able to induce non-MHC-restricted cytolysis of Daudi cells . All clones displayed a substantial reactivity to bacterial superantigens staphylococcal enterotoxin B and toxic shock syndrome toxin-1, irrespective of their gamma delta V gene usage . Furthermore, the gamma delta T cell clones expressed predominantly TCRDV2 and GV2 genes (26 of 35 clones), whereas the clones derived from CSF of MS patients expressed either DV1 or DV2 genes . The obtained gamma delta clones, in general, represented rather heterogeneous clonal origins, even though a predominant clonal origin was found in a set of 10 gamma delta clones derived from one patient with MS . The present study provides new evidence supporting a possible role of gamma delta T cells in the secondary inflammatory processes in MS.

J Immunol, 1995 May 1, 154(9), 4247 - 60
T cell receptor V alpha 4 is expressed by a subpopulation of V beta 6 T cells that respond to the bacterial superantigen staphylococcal enterotoxin B; Borrero H et al.; A subpopulation of murine minor lymphocyte-stimulating locus Ag (Mls)-1a-responsive, TCR V beta 6-expressing T hybrids was responsive to the superantigenic bacterial toxin, staphylococcal enterotoxin B (SEB), presented by murine MHC class II molecules . Comparative functional and surface marker analyses showed that this heterogeneity was not caused by nonspecific effects . It seemed, therefore, that the ability of the TCR V beta 6 T hybrids to respond to SEB was regulated by non-V beta TCR elements . cDNA sequencing analyses of the TCR beta- and alpha-chains expressed by the V beta 6 T hybrids and a beta-chain cDNA gene transfer experiment indicated that although J beta, CDR3 beta, J alpha, and CDR3 alpha were not the relevant elements, SEB responsiveness did correlate with the expression of two different members of the V alpha 4 family . A functional analysis of a separate panel of V alpha 4 T hybrids expressing different V beta elements, including V beta 6, specifically associated SEB responsiveness with the V alpha 4, V beta 6 combination . Overall, the data obtained with both panels of T hybrids showed SEB responsiveness in five out of five V beta 6, V alpha 4 T hybrids, and not in either 11 V beta 6 T hybrids expressing a variety of other V alpha elements or four V alpha 4 T hybrids expressing V beta 11, 14, and 15 elements . Thus, our experiments show for the first time a specific, positive qualitative effect of a defined V alpha element on responsiveness to a bacterial superantigen . Finally, this study has identified four new V alpha elements.

Clin Exp Dermatol, 1995 May, 20(3), 242 - 3
Troublesome myiasis complicated by peri-nephric abscess; Gibbs S; A normally fit European resident of East Africa developed a staphylococcal peri-nephric abscess following repeated episodes of furuncular myiasis caused by the tumbu fly, Cordylobia anthropophaga . A few lesions had required surgical intervention and one, presumably the source of sepsis, had become infected . There are very few descriptions in the literature of serious complications of myiasis and this is the first report of peri-nephric abscess from this source . This paper discusses the management of the myiasis and what factors determine the variation in host susceptibility and immune response to the infestation.

Biol Chem Hoppe Seyler, 1995 May, 376(5), 303 - 9
The domain structure and functional relationships in the bacterial superantigen, SEB; Hayball JD et al.; Staphylococcal enterotoxin B (SEB) has three essential biological properties that appear to be mediated by particular domains of the protein . As a superantigen, SEB triggers the T cell receptor (TcR) of a selected subset of T cells where the beta chain is encoded by particular V beta gene products . T cell triggering is generally dependent upon the ability of SEB to form a ternary complex with the TcR and MHC class II molecules and this interaction is relatively unrestricted by class II polymorphism . The third property of SEB is its potent toxicity; a few nanograms are sufficient to cause vomiting and violent diarrhoea in primates . In this study, we confirm the importance of the amino terminal domain of SEB for stimulation of human T cells . It is clear that the first 138 residues are the minimal mitogenic component of the toxin, and deletion of just seven more residues abrogates all activity . A mutant molecule with an internal deletion of these seven residues (SEB delta 131-138) was mitogenic suggesting that the region can confer stability to the rest of the protein, but does not include contact sites for either class II or the TcR . We further show that the disulphide loop is not essential for T cell recognition and that each of our mutants binds class II molecules with reduced affinity and is subtly variant in the pattern of TcR that it stimulates.

Toxicol Pathol, 1995 May-Jun, 23(3), 262 - 8
Aerosolized staphylococcal enterotoxin B-induced pulmonary lesions in rhesus monkeys (Macaca mulatta); Mattix ME et al.; The pathology of aerosolized staphylococcal enterotoxin B (SEB) was studied in the nonhuman primate . Six juvenile rhesus monkeys that received multiple lethal inhaled doses of SEB developed diarrhea and vomiting within 24 hr followed by depression, dyspnea, and shock . Three of 6 animals died by 52 hr . The most striking gross lesion in all 6 monkeys was diffuse severe pulmonary edema . Histologically, edema fluid was present within the peribronchiolar, peribronchial, and perivascular interstitium, alveolar septa, and alveoli . The adventitia of pulmonary vessels was infiltrated by lymphocytes, macrophages, and fewer neutrophils . Numerous large lymphocytes with occasional mitotic figures were within pulmonary vessels, often occluding alveolar capillaries . These cells were strongly immunoreactive with monoclonal antibodies against CD3, establishing them as T cells . Ultrastructurally, endothelial cell junctions were intact, and endothelial cells and type I pneumocytes contained numerous pinocytotic vesicles . Alveolar septal interstitial spaces were expanded by edema . The mechanism of these SEB-induced pulmonary lesions was not determined . We hypothesize that cytokine production by activated T cells may have caused vascular permeability changes leading to widespread pulmonary edema and shock.

J Heart Valve Dis, 1995 May, 4(3), 284 - 7
Experimental and clinical evaluation of formalin containing pericardial bioprostheses for replacement of infected valves; Garcia-Bengoechea JB et al.; In 1988 we quantified the adherence of two Staphylococcus epidermidis strains to glutaraldehyde-fixed bovine pericardium and teflon . The pericardium was used with and without prior rinsing to remove the preservative formaldehyde . Bacterial adherence was not detected on non-rinsed pericardium, and it was significantly less on rinsed pericardium than on teflon (p < 0.001) . In view of these results, we have since then implanted 31 pericardial bioprostheses without rinsing them in 29 patients with active bacterial endocarditis . Valve infection reoccurred in two patients (6.7%) and two patients developed early periprosthetic leaks in the absence of re-infection . No adverse clinical or biochemical findings attributable to residual aldehydes have been observed.

Glycobiology, 1995 May, 5(3), 327 - 33
Digalactosylceramide is the receptor for staphylococcal enterotoxin-B in human kidney proximal tubular cells; Chatterjee S et al.; We have characterized a glycosphingolipid (GSL) receptor for Staphylococcus enterotoxin-B (SEB) in cultured human kidney proximal tubular (PT) cells . Solid-phase binding of {125I}SEB to the GSL receptor was concentration dependent and was not displaceable by two structurally related toxins, such as staphylococcal enterotoxin-A and toxic shock syndrome toxin-1 . Rat kidney cells did not bind {125I}SEB . However, when the rat kidney cells were pre-incubated with digalactosylceramide, there was a concentration-dependent binding of {125I}SEB . Trimethylsilyl derivatization of methyl glycosides, followed by gas-liquid chromatography-mass spectrometry (GC-MS), revealed that galactose was the major sugar component of this putative receptor GSL . The sphingosines present in this GSL were d18:2, d22:2 and d23:0; the fatty acids present were palmitate, oleate and stearate . Permethylation of alditol acetates and GC-MS revealed two predominant sugars, namely 2, 3, 4 and 6 tetramethylgalactital and 2, 3 and 6 trimethylgalactital . The GSL receptor for SEB was sensitive to alpha-galactosidase, and resistant to beta-galactosidase and beta-glucosidase . Taken together, our studies reveal that the tentative structure of the receptor for SEB in human kidney PT cells is CerGal alpha 1-->4Gal . In summary, we have identified a GSL as one of the binding sites of SEB, a food-borne toxin . We believe that our finding may open up rational approaches for the therapy of SEB-induced glycopathology in man.

Acta Derm Venereol, 1995 May, 75(3), 218 - 21
In vitro T cell response to staphylococcal enterotoxin B superantigen in chronic plaque type psoriasis; Bour H et al.; Recent studies have demonstrated the important role of CD4+ T cells in the pathophysiology of psoriasis . One of the current hypotheses is that triggering of the psoriatic inflammatory process could be secondary to CD4+ T cell activation by bacterial superantigens in the skin . In this study, IL-2-derived T cell lines were recovered from the blood and the skin of 4 patients with chronic plaque type psoriasis and of 2 patients with allergic contact dermatitis (ACD) . Blood and skin T cell lines were tested for their ability to proliferate in vitro to staphylococcal enterotoxin B (SEB) presented by MHC class II expressing antigen-presenting cells . The results showed a significantly higher SEB-induced T cell proliferation in skin T cell lines as compared to blood T cell lines in 3 out of 4 psoriatic patients and in one of the 2 ACD patients . No difference between the skin and blood T cells for their response to phytohemagglutinin was observed . Furthermore the blood T cell lines from both patients and control individuals responded equally well to SEB . Thus psoriatic skin T cell lines were characterized by an enrichment in SEB-responding T cells . Since similar enhancement of SEB-responsive T cells was occasionally found in ACD patients, we propose that SEB could be an environmental factor associated with rather than responsible for psoriatic inflammation.

Immunology, 1995 May, 85(1), 57 - 62
Superantigens and conventional antigens induce different responses in alpha beta T-cell receptor transgenic mice; Blankson JN et al.; While superantigens such as staphylococcal enterotoxin B (SEB) have been shown to induce both clonal deletion and clonal anergy, it is still not known why tolerance rather than memory is induced . To address this issue, we tested the proliferative capacity of T cells from ovalbumin (OVA)-specific alpha beta T-cell receptor transgenic mice primed with either SEB emulsified in complete Freund's adjuvant (CFA) or with OVA peptide, the specific antigen, in CFA . By contrast cells from mice primed with SEB in CFA appeared to be anergic in that they were hyporesponsive to OVA peptide as well as to SEB . The anergic cells could respond to phorbol myristate acetate (PMA) and ionomycin, suggesting that a proximal signal transduction step was affected . Cells from transgenic mice primed with OVA peptide and CFA were not anergic and in fact displayed an enhanced response when they were challenged with OVA in vitro . Thus, when the two antigens are emulsified in CFA and then injected subcutaneously, they behave very differently: the superantigen SEB induces anergy whereas the conventional antigen OVA induces a memory type of response.

Clin Nucl Med, 1995 May, 20(5), 395 - 7
Tc-99m leukocyte scintigraphy in infective endocarditis; Adams BK; A 56-year-old man with Staphylococcal aureus septicemia and clinically suspected infective endocarditis was imaged using Tc-99m HMPAO leukocytes . Although abnormal uptake was seen at 3 hours after injection, 24-hour images demonstrated marked concentration of radiolabeled leukocytes in a myocardial abscess and large posterior parietal infarct . In addition, multiple splenic infarcts were shown . The Tc-99m labeled leukocyte scan proved to be an excellent whole body screening procedure for diagnosing sites of infection and infarction in this patient.

J Clin Microbiol, 1995 May, 33(5), 1385 - 8
Value of terminal subcultures for blood cultures monitored by BACTEC 9240; Shigei JT et al.; Blood cultures collected in BACTEC Plus Aerobic/F bottles and BACTEC Plus Anaerobic/F bottles were monitored for 5 days by BACTEC 9240 and subsequent terminal subcultures . Of the 13,471 bottles subcultured, 11.0% (1,477 of 13,471) were culture positive . Of these, 94.0% (1,388 of 1,477) were detected by BACTEC 9240; the additional 6.0% (89 of 1,477) were considered to be false negatives by BACTEC 9240 since they were detected by terminal subculture only . The false-negative bottles consisted of 17 BACTEC Plus Aerobic/F and 72 BACTEC Plus Anaerobic/F bottles, accounting for 2.2 (17 of 786) and 10.4% (72 of 691) of the total positive aerobic and anaerobic bottles, respectively . The positive blood culture bottles most frequently not detected by BACTEC 9240 grew Pseudomonas spp . (24), Staphylococcus spp . (21), and yeasts (24) . Of the 86 blood cultures represented by the 89 false-negative bottles, 41 would not have been identified as positive since the other bottle in the blood culture set was either a false negative or a true negative . In general, terminal subcultures of false-negative BACTEC bottles had heavy growth, indicating that BACTEC Plus media were able to support the growth of microorganisms, but the BACTEC 9240 instrument was unable to detect this growth.

Br J Surg, 1995 May, 82(5), 677 - 80
Influence of laparoscopic and conventional cholecystectomy upon cell-mediated immunity; Griffith JP et al.; Surgery, trauma and anaesthesia induce a state of transient immunosuppression . Laparoscopic cholecystectomy has several well documented clinical advantages over traditional cholecystectomy and provokes a lower acute phase response, thought to be a result of the smaller wound size . The influence of laparoscopic cholecystectomy (21 patients) and conventional open cholecystectomy (13 patients) upon components of the cell-mediated immune system was investigated . Cell-mediated immunity was studied by in vitro assays of T lymphocyte proliferation to different mitogens, and by natural killer cell cytotoxicity using a standard 51Cr release assay . Blood samples were taken before and 24 h after the start of the operation . In the sample taken after operation there was significant depression of T lymphocyte proliferation to phytohaemagglutinin (stimulation index 149.4 versus 33.3, P < 0.002), staphylococcal enterotoxin B (85.2 versus 52.6, P = 0.01) and toxic shock syndrome toxin (48.4 versus 14.8, P = 0.08) in the group of patients who underwent open surgery, but not in the group treated by laparoscopic surgery . There was a small but statistically insignificant decrease of natural killer cell cytotoxicity in both groups of patients . These findings suggest that laparoscopic cholecystectomy causes less depression of cell-mediated immunity than open cholecystectomy.

Int Arch Allergy Immunol, 1995 May-Jun, 107(1-3), 179 - 82
Staphylococcal enterotoxin B affects in vitro IgE synthesis, interferon-gamma, interleukin-4 and interleukin-5 production in atopic eczema; Neuber K et al.; Peripheral blood mononuclear cells from patients with atopic eczema (AE) stimulated with the 'superantigen' Staphylococcus enterotoxin B (SEB) secreted significantly more interleukin (IL)-4 and IL-5 as well as IgE, and markedly less interferon-gamma than those from healthy controls . Our results support the assumption that SEB produced by S . aureus colonizing the skin of patients with AE may induce expansion of IL-4- and IL-5-producing Th2 clones, leading to increased IgE synthesis and eosinophil activation.

Br J Ophthalmol, 1995 May, 79(5), 473 - 5
Rapid diagnosis of ocular herpes simplex infections; Asbell PA et al.; BACKGROUND--The Surecell herpes (HSV) test kit is a test for detecting the presence of herpes simplex viral antigen by means of a monoclonal antibody based immunoassay . The test has proved to be highly sensitive and specific in diagnosing genital, oral, and dermatological herpes infections . METHODS--In this study, samples from patients with ocular keratitis were evaluated by tissue cultures and the Surecell test . The eyes of New Zealand rabbits were then inoculated with HSV type 1 acute keratitis, acute Staphylococcus keratitis, and HSV type 1 postkeratitis (healed corneas) . Tear film samples collected from each eye with a cotton swab were evaluated by routine culture (A-549 monolayers) and by the Surecell test with and without prior placement of the swab in Hank's medium . RESULTS--The Surecell system had a 70% sensitivity and a 100% specificity in the detection of HSV antigen in ocular infections, and was shown to be a quick, efficient, and accurate method of testing for HSV antigen in humans . CONCLUSION--These results from humans and rabbits indicate that the Surecell test, which requires no special equipment, can be a useful in office adjunct in the clinical diagnosis of ocular herpes simplex.

Immunol Lett, 1995 May, 46(1-2), 9 - 14
Lethally irradiated normal strains of mice radioprotected with SCID bone marrow develop sensitivity to low doses of staphylococcal enterotoxin B; Aboud-Pirak E et al.; Normal strains of mice are rendered sensitive to small amounts (3-10 micrograms) of staphylococcal enterotoxin B (SEB) by transplanting bone marrow cells of SCID donor mice to lethally irradiated recipients . Four to 12 weeks post-transplantation, SEB induces 56-100% lethality . Transplantation of normal mouse bone marrow cells, either alone or with the SCID mouse selected bone marrow cells, does not confer SEB sensitivity . These data imply that either irradiation ablates certain cell population(s), that confer resistance to SEB in normal mice (populations that are absent in the SCID donor mice) or that the donor cells selectively repopulate recipients with SEB-sensitive cells . This model will help elucidate the cells, cytokines and the SEB peptide fragments responsible for SEB toxicity and will be useful in identifying promising vaccine candidates and in developing preventive medicines to protect against this potent toxin.

Eur Heart J, 1995 May, 16(5), 588 - 602
Identification of high-risk subgroups in infective endocarditis and the role of echocardiography; Erbel R et al.; The outcome of infective endocarditis remains poor . It has an overall mortality of around 30%, rising in high-risk subgroups to 50% and 100% . The prognosis can be improved by identification of high-risk patients and special management . Patients with infective endocarditis are found to be at high risk for death or serious complications when one or more of the following factors exist: old age (especially > 60 years old), delayed diagnosis, staphylococcal infection, aortic valve endocarditis, large valvular vegetation, congestive heart failure, embolization in the central nervous system or coronary artery, prosthetic valve infection, recurrent events, and failed antibiotic therapy . These factors often coexist and interrelate with one another . Early diagnosis and active treatment are critical for a better clinical outcome . However, infective endocarditis is difficult to diagnose because of the atypical clinical manifestations and frequent negative results from blood culture . Echocardiography plays an indispensable role in the diagnosis and management of suspected or known infective endocarditis . By detecting and monitoring certain pathological changes associated with the disease, e.g . vegetation, abscess formation, or valvular destruction, echocardiography helps to diagnose the disease early, to identify patients at high risk, to monitor the patients, and to optimize the timing and mode of surgical intervention . Serious complications can thus be avoided or cured at an early stage and the prognosis significantly improved.

Pediatr Pulmonol, 1995 May, 19(5), 256 - 61
Vitamin A status and airway infection in mechanically ventilated very-low-birth-weight neonates; Shenai JP et al.; Vitamin A (retinol) plays an important role in immunity . Respiratory and enteral infections in children are associated with low serum vitamin A concentrations that improve during recovery . To test the hypothesis that airway infection in very-low-birth-weight (VLBW) neonates likewise may be associated with a change in vitamin A status, we examined 20 VLBW neonates (selection criteria: birth weight 700-1300 g, gestational age 26-30 weeks, need for supplemental oxygen and mechanical ventilation for > 72 hr after birth) who were enrolled in the control group of a randomized clinical trial of vitamin A supplementation reported earlier . We studied changes in weekly measurements of plasma concentrations of vitamin A and retinol-binding protein (RBP) during 4 weeks following enrollment in the trial (postnatal day 4) and compared changes between periods with and without airway infections . Seventeen infants had 22 episodes of documented airway infection . Staphylococcus epidermidis was the predominant organism . Plasma vitamin A concentrations decreased during 19 out of 22 . With airway infection (mean change: -4.1 to -18.6 micrograms/dL), while they increased during 37 out of 58 periods without airway infection (mean change: -0.2 to +5.8 micrograms/dl; P < 0.001) . The mean (+/- SD) plasma vitamin A concentrations before, during, 1 week after, and 2 weeks after an episode of airway infection were 20.9 +/- 8.3, 9.7 +/- 4.1, 12.8 +/- 8.9, and 16.2 +/- 7.2 micrograms/dL, respectively . The mean value during airway infection was significantly lower than those before and two weeks after airway infection (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Mol Microbiol, 1995 May, 16(3), 477 - 84
Plasmid pT181 replication is decreased at high levels of RepC per plasmid copy; Iordanescu S; The replication of staphylococcal plasmid pT181 is indirectly controlled at the level of the synthesis of its replication initiator, RepC . As a result, high levels of RepC synthesis per plasmid copy were expected to lead to autocatalytic plasmid replication, which secondarily would affect host physiology . Surprisingly, RepC overexpression was found to lead to a rapid decrease in pT181 copy number and replication rate . These effects depended on the ratio of RepC to the pT181 replication origin rather than on the absolute amount of RepC in the cell . In a wild-type host, the increase in RepC/plasmid copy also inhibited chromosome replication and cell division . The changes in host physiology did not play any role in the decrease in pT181 replication caused by RepC overexpression since pT181 replication responded in the same way in a host mutant insensitive to the effects of RepC induction . These results suggest that pT181, the prototype of an entire class of plasmids from Gram-positive bacteria, responds to overexpression of its replication initiator by a decrease in plasmid replication.

Cell Immunol, 1995 May, 162(2), 315 - 20
Activation of murine T cells by bacterial superantigens requires B7-mediated costimulation; Muraille E et al.; Staphylococcus enterotoxins bind class II MHC molecules on antigen-presenting cells (APC) and stimulate T cells expressing appropriate V beta gene products . Although the role of non-TcR-associated costimulatory receptors during antigen-specific T cell stimulation has been clearly established, the involvement of costimulatory activity in T cell activation by superantigens (SAgs) has been the matter of controversy . The aim of this study was to evaluate the role of the costimulatory-receptor ligand molecules CD28/B7 on bacterial SAg-mediated activation of naive murine T cells . We demonstrate in this report that a combination of monoclonal antibodies to murine B7.1 and B7.2 molecules inhibits the in vitro response of naive T cells to SAgs SEA, SEB, and TSST-1 . The inhibition of T cell responses required simultaneous blocking of B7.1 and B7.2, suggesting that either B7.1 or B7.2 is sufficient to provide costimulatory signals to naive T cells in response to bacterial exotoxins . Inhibition of T cell activation by antibodies to B7-related molecules can be overcome by antibodies to CD28, a finding in agreement with the hypothesis that CD28-mediated signals participate in T cell activation by bacterial SAgs . These observations suggest that, as demonstrated for conventional antigen, T cell activation by SAgs requires the coordinated participation of TcR- and CD28-derived signals.

J Immunol, 1995 May 1, 154(9), 4395 - 403
Expression and function of mouse Fas antigen on immature and mature T cells; Nishimura Y et al.; We prepared mAbs specific for the mouse Fas Ag (CD95) and used them to analyze the expression and apoptosis-inducing activity of the Fas Ag on murine immunocytes . Cytofluorometry of mouse bone marrow, thymus, and splenocytes using the mAbs indicated that cells of the T lineage, except for bone marrow cells, expressed Fas Ag on the surface . CD4-CD8- undifferentiated thymocytes expressed low levels of Fas Ag . Immature CD4+CD8+ thymocytes and mature CD4+CD8- and CD4-CD8+ thymocytes were highly positive for Fas Ag . CD4+CD8+ thymocytes were specifically sensitive to the apoptosis-inducing activity of anti-Fas, although CD4-CD8-, CD4+CD8-, and CD4-CD8+ thymocytes were resistant . Spleen T cells were resistant to anti-Fas, whereas they expressed Fas Ag . The superantigen, staphylococcal enterotoxin B (SEB) administered to BALB/c mice, induced clonal expansion and successive clonal deletion of spleen T cells bearing the V beta 8 TCR, which specifically reacts to SEB . Such clonal deletion of V beta 8 T cells was highly suppressed in lpr mice, which have defects in the Fas Ag gene . In SEB-administrated BALB/c mice, expression of Fas Ag was significantly enhanced on V beta 8, but not on V beta 6 T cells, which cannot react to SEB . Moreover, V beta 8 T cells in SEB-primed mice were sensitive to the cell-killing activity of anti-Fas, although V beta 6 T cells were resistant . These findings show that the expression level and apoptosis-inducing activity of Fas Ag on peripheral T cells are directly up-regulated by stimulation through the TCR in vivo.

J Immunol, 1995 May 1, 154(9), 4302 - 8
Fas ligand-mediated cytotoxicity is directly responsible for apoptosis of normal CD4+ T cells responding to a bacterial superantigen; Ettinger R et al.; Exposure of naive CD4+ T lymphocytes to superantigens such as staphylococcal enterotoxin B (SEB) induces a strong proliferative response . Prolonged exposure or subsequent restimulation of the responding T cell population with SEB leads to the apoptotic events of activation-induced cell death (AICD) . However, T cells derived from either Fas-deficient lpr or Fas ligand-deficient gld autoimmune mouse strains, fail to undergo AICD under these conditions . Instead, these autoimmune T cells mount a vigorous proliferative response, suggesting a critical role for Fas/FasL interactions in this form of autoapoptosis . In the current study, we found that SEB-induced AICD was tied to the rapid induction of FasL expression in cells constitutively expressing high levels of Fas . Furthermore, the addition of soluble Fas-IgG fusion protein to the SEB-restimulated cultures blocked AICD and resulted in a 2 degrees proliferative response that was comparable in magnitude and kinetics to that of the lpr and gld T cells . The rapid onset of apoptosis in normal T cells subsequent to restimulation with SEB was in direct contrast to the proliferative response of the initial cultures, even though comparable levels of Fas and FasL RNA were found in T cells after 1 degree and 2 degrees challenge . The clonal expansion of the normal T cells responding to the initial SEB stimulation was, however, dramatically compromised when the normal cells were cocultured with an MRL-lpr responder population; addition of soluble Fas-IgG rescued the normal component of the response . Together, these data demonstrate first, that Fas/FasL interactions are intimately tied to superantigen-induced AICD, a form of autocrine cell death, and second, that FasL-mediated cytotoxicity is responsible for the disappearance of normal CD4+ T cells in lpr cocultures.

Philos Trans R Soc Lond B Biol Sci, 1995 Apr 29, 348(1323), 27 - 34
Initial studies of the equilibrium folding pathway of staphylococcal nuclease; Wang Y et al.; Spectroscopic methods were used to examine the sequential build up of structure in the denatured state of staphylococcal nuclease . The 'free energy distance' between the native and denatured states was manipulated by altering conditions in solution (for example altering urea or glycerol concentration) and by changing the amino acid sequences . Initial studies employed a fragment of nuclease, referred to as delta 131 delta, which lacks six structural residues from the amino terminus and one structural residue from the carboxy-terminus . Nuclear magnetic resonance analysis of this fragment in solution revealed a modest quantity of dynamic structure which is native-like in character . With the addition of urea, 12 new HN peaks appeared in the 1H-15N correlation spectrum, presumably as a result of the breakdown of residual structure involving the first three beta strands . With the addition of glycerol, there was a rapid increase in the quantity of beta sheet structure detected by circular dichroism spectroscopy . At very high glycerol concentrations, an increase in helical structure became apparent . These data in addition to previously published results suggest that: (i) a beta-meander (strands beta 1-beta 2-beta 3) and the second alpha helix (alpha 2) are among the most stable local structures; (ii) the five-strand beta-barrel forms in a reaction which does not require the presence of several other native substructures; and (iii) the last step on the equilibrium folding pathway may be the formation and packing of the carboxy terminal alpha helix (alpha 3) to give the native state.

J Immunol Methods, 1995 Apr 26, 181(2), 233 - 43
Whole blood culture for measuring mitogen induced T cell proliferation provides superior correlations with disease state and T cell phenotype in asymptomatic HIV-infected subjects; Bocchieri MH et al.; Proliferative responses to a panel of mitogens were compared in parallel for two sources of cells, whole blood (WB) and conventionally prepared peripheral blood mononuclear cells (PBMC), obtained from asymptomatic HIV seropositive and control subjects . Weak but statistically significant correlations of the proliferative responses were observed . Use of either lymphocyte source produced significant differences in the proliferative responses between the HIV seropositive and control subjects, but the use of WB was more powerful, with a smaller sample size being required to discriminate between the proliferative responses of the two study groups . Furthermore, proliferative responses using WB gave strong and highly significant correlations with a number of important changes in the surface marker phenotype of the lymphocyte populations in the HIV seropositive subjects including CD4, CD8, CD4:CD8 ratio and certain CD8 subsets, whereas strong correlations were not observed with the PBMC . The response of WB lymphocytes to staphylococcal enterotoxin B (SEB) was highly reproducible and provided the best discrimination between HIV-infected and control subjects . We conclude that the use of WB for measuring lymphoproliferation is easy, rapid, accurate, and discriminative for assessing and following the changes in immune function which occur in HIV seropositive subjects, applicable in the clinical as well as in the research setting.

Science, 1995 Apr 21, 268(5209), 403 - 5
Superantigen-dependent, cell-mediated cytotoxicity inhibited by MHC class I receptors on T lymphocytes; Phillips JH et al.; Bacterial superantigens bind with high affinity to major histocompatibility complex (MHC) class II antigens on antigen-presenting cells and with T cell antigen receptor (TCR) beta chains on T lymphocytes, which results in the T cell activation responsible for toxic shock syndrome and food poisoning . Many cytotoxic T lymphocyte (CTL) clones were shown to have receptors for human leukocyte antigen (HLA) class I molecules that inhibited superantigen-induced cytotoxicity against appropriate class I-bearing target cells . One type of inhibitory receptor, NKB1, was present on CD4+ and CD8+TCR alpha beta+ CTL clones and blocked the killing of staphylococcal enterotoxin B (SEB)-coated targets bearing certain polymorphic HLA-B molecules . Expression of HLA-A, -B, and -C molecules on the SEB-coated targets also protected against cytolysis mediated by many NKB1-negative T cell clones, suggesting the presence of additional inhibitory MHC class I receptors . These HLA class I receptors may limit tissue destruction and possibly autoimmunity caused by activated T lymphocytes.

EMBO J, 1995 Apr 18, 14(8), 1607 - 14
Potassium-inhibited processing of IL-1 beta in human monocytes; Walev I et al.; Agents that deplete cells of K+ without grossly disrupting the plasma membrane were found to stimulate the cleavage of pro-interleukin (IL)-1 beta to mature IL-1 beta . Agents examined in this study included staphylococcal alpha-toxin and gramicidin, both of which selectively permeabilize plasma membranes for monovalent ions, the ionophores nigericin and valinomycin, and the Na+/K+ ATPase inhibitor ouabain . K+ depletion by brief hypotonic shock also triggered processing of pro-IL-1 beta . The central role of K+ depletion for inducing IL-1 beta maturation was demonstrated in cells permeabilized with alpha-toxin: processing of pro-IL-1 beta was totally blocked when cells were suspended in medium that contained high K+, but could be induced by replacing extracellular K+ with Na+, choline+ or sucrose . To test whether K+ flux might also be important in physiological situations, monocytes were stimulated with lipopolysaccharide (LPS) for 1-2 h to trigger pro-IL-1 beta synthesis, and transferred to K(+)-rich medium . This maneuver totally suppressed IL-1 beta maturation . Even after 16 h, however, removal of K+ from the medium resulted in rapid processing and export of IL-1 beta . Ongoing export of mature IL-1 beta from cells stimulated with LPS for 2-6 h could also be arrested by transfer to K(+)-rich medium . Moreover, a combination of two K+ channel blockers inhibited processing of IL-1 beta in LPS-stimulated monocytes . We hypothesize that K+ movement and local K+ concentrations directly or indirectly influence the action of interleukin-1 beta-converting enzyme (ICE) and, possibly, of related intracellular proteases.

Biochemistry, 1995 Apr 18, 34(15), 4909 - 12
Evidence for a molten globule-like transition state in protein folding from determination of activation volumes; Vidugiris GJ et al.; One of the most important, yet elusive, aspects of the protein folding question lies in the nature of the transition state . Direct information about the structural properties of the transition state can be obtained from determination of the activation volumes for the folding and unfolding transitions . The present pressure-jump relaxation study on the folding/unfolding of staphylococcal nuclease reveals that the volume of the protein-solvent system is larger in the transition state than in either the folded or unfolded states . Moreover, the activation volume of folding is much larger than that of unfolding . These results support a molten globule-like model for the transition state of nuclease in which the polypeptide chain is in a collapsed, loosely packed, solvent-excluded structure . In this model, hydrophobic collapse with concomitant desolvation is the rate-limiting step in the folding of the polypeptide chain, and solvent-excluded expansion of the folded state is the rate-limiting step in protein unfolding.

FEMS Microbiol Lett, 1995 Apr 15, 128(1), 39 - 44
Typing of Staphylococcus epidermidis strains by random amplification of polymorphic DNA; Marquet-Van Der Mee N et al.; The polymerase chain reaction was used to obtain randomly amplified polymorphic DNA profiles for typing of Staphylococcus epidermidis strains . Epidemiologically unrelated S . epidermidis isolates were screened with randomly amplified polymorphic DNA analysis . The discriminating ability of 45 randomly designed 10-mer primers was assessed . The highest discriminatory power was obtained with the 10-mer oligonucleotide OPAM-12 . In typing a total of 13 unrelated S . epidermidis strains with OPAM-12, 11 different banding profiles were obtained reproducibly by agarose gel electrophoresis . The discriminatory power of the method with OPAM-12 was estimated using the D value of Hunter and Gaston (1988) to be 0.961 . A reproducibility index of 1 was obtained after typing a total of 40 cultures including 12 triplicates and one quadruplicate of the 13 unrelated strains . Following the described procedure, the randomly amplified polymorphic DNA method provided a rapid, simple and reproducible alternative to other S . epidermidis typing systems.

Cell Immunol, 1995 Apr 15, 162(1), 26 - 32
Staphylococcal enterotoxin B induces the expression of activation markers on murine memory T cells in the absence of proliferation or lymphokine secretion; Lee WT et al.; Superantigens have been used to study peripheral tolerance in CD4+ T cells . The superantigen SEB induces T cell anergy by promoting the differentiation of SEB-activated virgin T cells into anergic memory T cells . Memory T cells from SEB or antigen-primed mice do not proliferate when they are cultured with SEB . The present studies were performed to determine whether memory T cells fail to interact with SEB antigen-presenting cells or whether SEB promotes incomplete or negative signals in memory T cells . When murine virgin and memory T cells were separated on the basis of CD45RB expression and cultured with SEB-pulsed B cells, SEB induced the expression of CD25, which then mediated proliferation when IL-2 was added to the cultures . In addition, SEB promoted the expression of the CD40L, which is required for T helper cell function . Finally, PMA induced a costimulatory signal leading to the proliferation of these cells . Surprisingly, the agents, i.e., IL-2 and PMA, which induced TM cell proliferation in conjunction with SEB failed to induce lymphokine secretion . However, in the presence of IL-4 plus IL-5, the T memory cells induced the SEB-pulsed B cells to secrete IgM and IgG . These results suggest that memory T cells are not simply unresponsive to SEB but are actively anergized.

Proc Natl Acad Sci U S A, 1995 Apr 11, 92(8), 3611 - 5
Differential modulation of Th1- and Th2-related cytokine mRNA expression by a synthetic peptide homologous to a conserved domain within retroviral envelope protein; Haraguchi S et al.; The influence of a synthetic retroviral peptide, CKS-17, on T helper type 1 (Th1)- or Th2-related cytokines was investigated in human blood mononuclear cells . Cells were stimulated with staphylococcal enterotoxin A, anti-CD3 plus anti-CD28 monoclonal antibodies, or lipopolysaccharide to induce cytokine mRNA . mRNA was detected by a reverse transcription-polymerase chain reaction or Northern blot analysis . CKS-17 down-regulated stimulant-induced mRNA accumulation for interferon gamma (IFN-gamma), interleukin (IL)-2, and p40 heavy and p35 light chains of IL-12, a cytokine that mediates development of Th1 response . CKS-17 up-regulated stimulant-induced mRNA accumulation of IL-10 and did not suppress Th2-related cytokine (IL-4, IL-5, IL-6, or IL-13) mRNA expression . A reverse sequence of CKS-17 peptide, used as a control, showed no such action . Anti-human IL-10 monoclonal antibody blocked ability of CKS-17 to inhibit mRNA accumulation for IFN-gamma but not the CKS-17 suppressive activity of IL-12 p40 heavy chain mRNA . Thus, CKS-17-mediated suppression of IFN-gamma mRNA expression is dependent upon augmentation of IL-10 production by CKS-17 . This conserved component of several retroviral envelope proteins, CKS-17, may act as an immunomodulatory epitope responsible for cytokine dysregulation that leads to suppression of cellular immunity.

J Biol Chem, 1995 Apr 7, 270(14), 7799 - 808
Further characterization of HeLa DNA polymerase epsilon; Chui G et al.; DNA polymerase epsilon (pol epsilon) from HeLa cells was purified to near homogeneity, utilizing Mono S fast protein liquid chromatography for complete separation from pol alpha . The purified pol epsilon preparation showed two polypeptides of > 200 and 55 kDa and a small amount of active 122-kDa proteolysis product on denaturing polyacrylamide gels . Pol epsilon (as well as pols alpha and delta) is optimally active in 100-150 mM potassium glutamate and 15 mM MgCl2 . Replication factors RF-A and RF-C, proliferating cell nuclear antigen, and Escherichia coli single-stranded DNA binding protein showed no significant effect on this preparation's pol epsilon activity, processivity, or substrate specificity . The size of the pol epsilon transcript for the catalytic subunit (> 200 kDa) was investigated in both normal human fibroblasts and HeLa cells . A 7.7-kilobase transcript was detected which was 5-16-fold more prevalent in proliferating than in quiescent HeLa cells . No significant difference in the level of pol epsilon transcript in HeLa cells or fibroblasts was seen after ultraviolet irradiation . Mouse polyclonal antiserum was produced to a 144-amino acid fragment of pol epsilon fused to staphylococcal protein A . This non-neutralizing polyclonal antiserum specifically recognized the catalytic subunit of pol epsilon by immunoblotting, but not that of pol alpha, beta, or delta . In addition, mouse polyclonal antiserum raised against column-purified pol epsilon was able to recognize and to neutralize pol epsilon, and a mouse monoclonal antibody was raised which was able to recognize specifically the catalytic subunit of pol epsilon.

Biochim Biophys Acta, 1995 Apr 5, 1248(1), 27 - 34
Identification of the active site histidine in Staphylococcus hyicus lipase using chemical modification and mass spectrometry; Boots JW et al.; Staphylococcus hyicus lipase is a serine hydrolase . In order to identify the active site histidine of S . hyicus lipase we have chemically modified S . hyicus lipase with 1-bromo-octan-2-one . The enzyme is rapidly inactivated by this inhibitor with a half-time of 578 s at pH 6.5 and 30 degrees C . Addition of the enzyme's cofactor calcium increases the inactivation rate approx . 2-fold . When n-hexadecylphosphocholine, a non-hydrolysable substrate analogue, is added the inactivation rate decreases about 3-fold, suggesting that a residue in the active site of S . hyicus lipase is involved in the inactivation reaction . Inactivation of S . hyicus lipase with 14C-labelled 1-bromo-octan-2-one shows that 1.4 moles of inhibitor per mole of lipase are incorporated . The results of an electrospray mass spectrometric study of the inactivated enzyme are consistent with this finding . In order to identify the modified residue, both the inactivated and the unmodified lipase were digested with cyanogen bromide followed by trypsin . The resulting peptides were analysed using HPLC and fast atom bombardment mass spectrometry . The results allow the modified residue to be assigned to the peptide Gly597-Lys612 . Collision induced dissociation mass spectrometry allowed the modified residue to be identified as His-600 . From these results we conclude that this residue forms part of the catalytic triad of S . hyicus lipase.

Biochemistry, 1995 Apr 4, 34(13), 4316 - 24
X-ray crystal structures of staphylococcal nuclease complexed with the competitive inhibitor cobalt(II) and nucleotide; Loll PJ et al.; Two crystal structures of ternary complexes of staphylococcal nuclease, cobalt(II), and the mononucleotide pdTp are reported . The first has been refined at 1.7 A to a crystallographic R value of 0.198; the second, determined from a crystal soaked for 9 months in a slightly different mother liquor than the first crystal, has been refined at 1.85 A to an R value of 0.174 . In the first structure, the cobalt ion is displaced 1.94 A from the normal calcium position, and the active site is dominated by a salt bridge between Asp-21 and Lys-70 from a symmetry-related molecule in the crystal lattice . The Co2+ ion appears unable to displace this lysine; consequently, the metal is bound in a vestibular site adjacent to the calcium site . The metal-binding pocket in the second structure adopts a configuration similar to that of the calcium complex, with the cobalt ion binding only 0.36 A from the calcium position . However, an inner sphere water seen in the calcium structure is missing from this structure . The cobalt ion in the second structure appears to be loosely or transiently coordinated within the calcium binding pocket, as evidenced by the high value of its refined thermal factor . Loss of catalytic activity for cobalt(II)-substituted nuclease is perhaps due to its inability to bind this inner sphere water.

Nat Immun, 1995 Apr, 14(4), 188 - 97
Human intestinal V delta 1+ T cells obtained from patients with colon cancer respond exclusively to SEB but not to SEA; Maeurer M et al.; The function and activation requirements for gamma delta T cells residing in the human intestine are still poorly defined . We have established two gamma delta + T cell tumor-infiltrating lymphocyte (TIL) lines derived from a primary colorectal cancer (gamma delta TIL No . 3481), and from a colorectal cancer lesion metastatic to the liver (gamma delta TIL No . 7279) . Both gamma delta TIL lines used exclusively the V delta 1 segment and predominantly the V gamma 2 segments of the T cell receptor (TCR) variable regions and lysed allogeneic colorectal cancer cell lines, e.g . HCT 116, but not natural killer/lymphokine-activated-killer-sensitive target cell lines, e.g . K562 or Daudi . gamma delta T cell effector functions were evaluated on the basis of their recognition and cytolysis of colorectal cancer cell lines, T cell proliferation, and interferon (IFN)-gamma release . Both gamma delta T cell lines exhibited similar responses to the staphylococcal superantigens (SE) A and B . SEA and SEB did not influence target cell cytolysis of colon cancer targets . Neither gamma delta + T cell line responded to SEA as measured by IFN-gamma release of T cell proliferation . In marked contrast, SEB induced T cell proliferation and IFN-gamma release in the absence of stimulator cells . SEB induced secretion of IFN-gamma by gamma delta T cells which could be augmented if stimulator cells (HCT116) were also added to gamma delta T cells . On the basis of these data, we suggest that intestine-derived V delta 1/V gamma 2+ T cells respond preferentially to SEB and not to SEA . This disparity may reflect the inherently higher affinity of individual gamma delta TCR subsets for SEB but not to SEA and/or indicate that a subset of gamma delta + TILs in patients with colon cancer may be preferentially expanded with a TCR rearrangement favoring the interaction with SEB . The induction of IFN-gamma release and proliferative gamma delta + T cell responses by SEB suggests a pivotal role for intestinal gamma delta T cells in mediating immune responses against bacteria and bacterial products, or potentially in anti-tumor-directed immunity . Such immune responses mediated by gamma delta + T cells may take place prior to the maturation of antigen-specific MHC-restricted alpha beta + T cell responses.

Bioorg Med Chem, 1995 Apr, 3(4), 437 - 45
Targeting peptide nucleic acid-protein conjugates to structural features within duplex DNA; Norton JC et al.; A convenient small scale synthesis has been developed for obtaining peptide nucleic acid oligomers (PNAs) . PNAs have been conjugated to a protein, staphylococcal nuclease, through disulfide exchange between a cysteine at the 3'-(carboxy) end of the PNA and an introduced cysteine on the surface of the nuclease . Site specific DNA cleavage by the attached nuclease has been used to examine the Watson-Crick hybridization of the PNAs to duplex DNA . Substantial affinity cleavage occurred when target sites contained inverted repeats which have the potential to form non B-DNA structures such as cruciforms . No affinity cleavage was observed at a site lacking apparent potential for non B-DNA structures . These results indicate that the Watson-Crick hybridization of PNAs to duplex DNA by strand displacement is favored by the presence of potential alternative secondary structures within the target sequence.

Infect Immun, 1995 Apr, 63(4), 1223 - 8
In vitro modulation of keratinocyte-derived interleukin-1 alpha (IL-1 alpha) and peripheral blood mononuclear cell-derived IL-1 beta release in response to cutaneous commensal microorganisms; Walters CE et al.; The ability of a range of skin commensal microorganisms to modulate interleukin-1 (IL-1) release by cultured human keratinocytes and peripheral blood mononuclear cells (PBMCs) was investigated by a combination of enzyme-linked immunosorbent assays and bioassays . Three fractions (formaldehyde-treated whole cells, culture supernatants, and cellular fractions) were prepared from Propionibacterium acnes, Propionibacterium granulosum, Staphylococcus epidermidis, Staphylococcus capitis, Staphylococcus hominis, and Malassezia furfur serovar B . The levels of immunochemical IL-1 alpha released by cultured keratinocytes during coincubations with these microbial fractions ranged from 0 to 136 pg/ml and were maximal after 72 h . No microbial fraction consistently upregulated immunochemical IL-1 alpha release by freshly isolated keratinocytes from two donors and a transformed cell line, all of which produced the cytokine constitutively to various extents . Bioassays revealed that most of the IL-1 released was biologically inactive . In contrast, whole cells of formaldehyde-treated P . granulosum and S . epidermidis significantly stimulated release of IL-1 beta by PBMCs from three donors compared with the negative control (culture medium) . Release was maximal at 24 h . Coincubation with intact cells of the yeast M . furfur significantly decreased levels of IL-1 beta below the values for the negative control by PBMCs from all three donors . There was good correlation between bioassay data and immunoassay data for IL-1 beta, and the depressive effect of M . furfur cells on cytokine production by all three cultures of PBMCs was mirrored in the levels of bioactive cytokine . This reduction in IL-1 beta release by PBMCs by M . furfur may provide an explanation why dermatoses thought to be caused by this yeast are essentially noninflammatory or only mildly inflammatory.

Infect Immun, 1995 Apr, 63(4), 1158 - 64
Anti-gamma interferon and anti-interleukin-6 antibodies affect staphylococcal enterotoxin B-induced weight loss, hypoglycemia, and cytokine release in D-galactosamine-sensitized and unsensitized mice; Matthys P et al.; Administration of staphylococcal enterotoxin B (SEB) to BALB/c mice was found to induce a cytokine release syndrome hallmarked by weight loss and hypoglycemia . A neutralizing monoclonal antibody against gamma interferon (IFN-gamma) given before SEB counteracted weight loss and prevented hypoglycemia . This protective effect of anti-IFN-gamma antibody was associated with decreased IFN-gamma levels in serum; tumor necrosis factor (TNF) and interleukin-6 (IL-6) levels remained unchanged . A monoclonal anti-IL-6 antibody, known for its ability to cause accumulation of biologically active IL-6 in the circulation, did not modify SEB-induced body weight loss or hypoglycemia . Levels of TNF, IFN-gamma, and IL-6 in serum were all more elevated in anti-IL-6-treated mice than in corresponding SEB-challenged control mice . In D-galactosamine-sensitized mice, SEB-induced weight loss but not hypoglycemia was more severe, resulting mostly in death within 24 h . Higher levels of biologically active TNF and IFN-gamma in serum were noted in these mice than in mice receiving SEB only . In D-galactosamine-sensitized mice, anti-IFN-gamma antibody did prevent hypoglycemia but failed to reduce the severity of the syndrome . Again, TNF levels in anti-IFN-gamma-treated mice remained unchanged . Pretreatment with anti-IL-6 antibody temporarily attenuated SEB-induced hypoglycemia in sensitized mice . Thus, at 6 h post-SEB injection, anti-IL-6-treated mice were less hypoglycemic than corresponding controls . However, at 24 h, hypoglycemia was significantly aggravated . Concomitantly, IL-6 levels were dramatically increased . Neither anti-IFN-gamma nor anti-IL-6 antibody treatment modulated mortality levels in D-galactosamine-sensitized mice . The data obtained with anti-IFN-gamma antibody clearly indicate that endogenous IFN-gamma is instrumental in bringing about hypoglycemia and body weight loss in mice exposed to SEB but also that hypoglycemia is not a crucial determinant of mortality in D-galactosamine-sensitized mice . The data obtained with anti-IL-6 antibody indicate that endogenous IL-6 is involved in regulating the levels of TNF and IFN-gamma in serum.

Immunology, 1995 Apr, 84(4), 528 - 35
Superantigen-induced peripheral T-cell deletion: the effects of chemical modification of antigen-presenting cells, interleukin-4 and glucocorticoid hormones; Ayroldi E et al.; Experiments were performed to evaluate the role of antigen-presenting cells (APC) and the effect of interleukin-4 (IL-4) and glucocorticoid hormone (GCH) exposure on the in vitro deletion of CD4+ CD8- and CD8+ CD4- T cells by staphylococcal enterotoxin B (SEB) . APC fixation with the chemical cross-linker 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide (ECDI) inhibited their capacity to induce SEB-specific deletion of mature T lymphocytes . Deletion was not influenced by treatment with anti-CD28 antibodies, which modulate T-cell activation . However, it was augmented by IL-4, known to counteract anti-CD3- and GCH-induced thymocyte apoptosis, and was inhibited by dexamethasone (DEX) . These results indicate that metabolically active APC are required for deletion of antigen-specific mature T cells and suggest that IL-4 and GCH can modulate this phenomenon in vitro.

Antimicrob Agents Chemother, 1995 Apr, 39(4), 834 - 8
Tolerability, kinetics, and efficacy of subconjunctival pefloxacin in pigmented rabbits; Marrakchi-Benjaafar S et al.; Pefloxacin has been shown to have good intraocular penetration when given systemically . In order to extend its clinical use, we have assessed the tolerability, kinetics, and efficacy of subconjunctival pefloxacin in phakic pigmented rabbits . The tolerability of a single subconjunctival injection of pefloxacin (0.8, 1.6, 8, or 16 mg in 0.2 ml) in the right eyes of eight rabbits was evaluated by clinical and histopathological examination . The 0.8-mg dose of pefloxacin was well tolerated . The kinetics was evaluated after a single subconjunctival injection of 0.8 mg in 18 rabbits . Animals were sacrificed at 1, 3, 5, 7, 12, or 18 h postinjection . Drug concentrations were measured by high-performance liquid chromatography . Pefloxacin was found in the cornea (maximum concentration, 18.13 micrograms/ml; half-life, 3.92 h) and in the aqueous humor (maximum concentration, 3.40 micrograms/ml; half-life, 2.14 h) . Pefloxacin did not penetrate into the vitreous humor by this route . The efficacy was evaluated in an experimental model of staphylococcal corneal ulcers in eight rabbits which received two subconjunctival injections of 0.8 mg of pefloxacin at 16 and 24 h after intrastromal inoculation . The results (expressed as mean log10 CFU per milliliter +/- standard deviation) showed that pefloxacin significantly (P < 0.001) reduced the bacterial counts (4.39 +/- 0.97) compared with those in control eyes (6.46 +/- 0.69) . For phakic eyes, subconjunctival pefloxacin might be of value for the treatment of corneal ulcers . Further studies are required to determine its penetration into the vitreous humor of aphakic eyes.

Br J Haematol, 1995 Apr, 89(4), 698 - 703
In vitro inhibition of tumour necrosis factor-alpha and interleukin-6 production by intravenous immunoglobulins; Toungouz M et al.; In vitro data about the action of pooled immunoglobulins (Ig) on cytokine (CK) production are controversial . The recent finding of natural antibodies against staphylococcal toxins neutralizing superantigen-induced activation prompted us to design an assay determining their ability to modulate staphylococcal enterotoxin B (SEB) induced CK production (IL-6 and TNF-alpha) . Presence of anti-SEB antibodies was demonstrated by a dot-blot assay in the three preparations tested . Preincubation of SEB with pooled Ig prior to addition into the test tube containing PBMCs (neutralizing condition) resulted in a strong inhibition of both TNF-alpha and IL-6 release (TNF alpha: 59 +/- 5% inhibition, P < 0.0001; IL-6: 71 +/- 7% inhibition, P < 0.0001, n = 15) . Anti-CD3 MoAb-induced CK production was not modified . During our study it was found that experimental conditions were critical to observe this inhibitory effect . Reversing the previous procedure by adding PBMCs into the test tube containing pooled Ig mixed with SEB resulted in a marked induction of TNF-alpha and IL-6 production . The same observation was made when pooled Ig solely was added (coating condition) . F(ab')2 fragments of pooled Ig displayed similar inhibitory capacity when added in neutralizing condition, indicating that the mechanism involved was not Fc dependent . The fragments lost the activating properties of intact Ig when incubated in coating condition, showing that Fc receptor activation occurs in this setting . The present work demonstrates that inhibition of SEB-induced CKs release by pooled Ig can be achieved by SEB neutralization, provided that the experimental conditions avoid activation through the Fc receptor . It can be assumed that similar mechanisms take place in some clinical conditions during which pooled Ig are infused.

Ann Rheum Dis, 1995 Apr, 54(4), 298 - 304
Staphylococcal enterotoxin B increases the severity of type II collagen induced arthritis in mice; Wooley PH et al.; OBJECTIVE--To observe the influence of T cell subset changes on the development of experimental arthritis, by using the bacterial superantigen staphylococcal enterotoxin B (SEB) to modulate the T cell repertoire during the arthritogenic response to type II collagen (CII) in vivo . METHODS--DBA/1 mice were injected with SEB before immunisation with CII, and assessed for the development of collagen induced arthritis (CIA) and an immune response to CII . Mice with established arthritis were also treated therapeutically with SEB . Flow cytometry was used to evaluate the effect of the therapy on T cell subsets and T cell receptor (TCR) V beta expression . RESULTS--Mice injected with SEB developed arthritis significantly faster than saline treated control animals, and developed more severe clinical features . Mice treated with SEB after the onset of CIA were also observed to progress more rapidly to a severe arthritis than mice treated with saline alone . The level of anti-CII antibody was not affected by SEB injection . Flow cytometric analysis of TCR expression in mice 21 days after injection of CII showed decreased expression of V beta 6 and V beta 8 cells in SEB treated mice, compared with collagen immunised control mice . Injection of SEB alone caused a decrease in V beta 8, but not V beta 6 T cells compared with the values in normal DBA/1 mice . No significant variations in the T cell repertoire were detected 70 days after CII immunisation . CONCLUSIONS--Treatment with the bacterial enterotoxin SEB before the induction of arthritis did not suppress the immunological or arthritogenic response to CII in DBA/1 mice, despite the modulation of the V beta 8 T cell subset . Treatment of mice with established arthritis using SEB provoked a more severe disease course.

Arch Dis Child, 1995 Apr, 72(4), 325 - 9
Morbidity using subcutaneous ports and efficacy of vancomycin flushing in cancer; Rubie H et al.; An evaluation of totally implanted venous access systems inserted in 163 consecutive children with cancer is reported . From 1988 to 1994, 180 subcutaneous ports were inserted in children more than 1 year old . Initial diagnosis was acute leukaemia (n = 79), non-Hodgkin's lymphoma (n = 33), and solid tumour (n = 51) . Median age was 85 months . All venous procedures were performed through the device . Chemotherapy was either moderate (n = 13) or intensive (n = 119) or very intensive (n = 48), including 16 patients undergoing marrow transplantation . Cumulative venous access totalled 55,770 patient days with a mean of 305 days/subcutaneous port . The cause of device removal was, end of treatment (n = 111), death due to malignancy (n = 20), catheter related infection (n = 7), and occlusion of the system (n = 4) . Mechanical complications occurred in 19 ports; 16 were due to clots, of which 14 were cleared with instillation of urokinase . Documented infectious episodes occurred in 47 ports, recurred once in 14, and twice in five cases . Among these infections, 47 were septicaemic; 31 due to Staphylococcus epidermidis . Twenty seven of initial septic episodes were considered to be catheter related; the rate was 15%/subcutaneous port or 0.05/100 catheter days . Risk factors for the development of a first infection were age below 4 years and the time of use . Since February 1993, vancomycin (50 micrograms/ml) has been given and this has reduced the rate of S epidermidis infection from 26/83 subcutaneous port to 4/97 . Life table analysis showed that the infection free interval for staphylococcus was significantly better after this technique ws initiated (log rank rest=0.02) . Time saved was approximately 30minutes/patient/week compared with external catheters, or 45 hours/month for the cohort of children treated . Subcutaneous ports in paediatric cancer patients are reliable, safe, and durable and may offer an attractive alternative to external catheters for prolonged venous access and intensive treatment.

J Neurol Neurosurg Psychiatry, 1995 Apr, 58(4), 444 - 6
Comparison of rates of infection of two methods of emergency ventricular drainage; Kim DK et al.; The rates of infection of two methods of external ventricular drainage in use at Atkinson Morley's Hospital--namely, (a) percutaneous drainage with Rickham reservoirs and (b) tunnelled ventriculostomies--were compared in this retrospective review . Percutaneous drainage of CSF with Rickham reservoirs was associated with a 27% rate of infection as identified by positive microbiological cultures; tunnelled ventriculostomy catheters had a 10% infection rate . The difference in the infection rate between the two methods was statistically significant (P < 0.015) . Other variables examined, including the age and sex of the patients and the reasons for ventricular drainage, were not associated with an increased rate of infection . Most infections from either method were caused by a coagulase negative staphylococcus . The average duration of ventricular drainage before identification of positive cultures was 5.7 days for Rickham reservoirs and 6.0 days for ventriculostomies.

Nutr Clin Pract, 1995 Apr, 10(2), 60 - 6
Femoral catheters increase risk of infection in total parenteral nutrition patients; Harden JL et al.; Central venous access for the administration of total parenteral nutrition is usually achieved via the subclavian or internal jugular veins . Although a high incidence of complications has been reported with the use of femoral catheters for central venous access, this route has been used when traditional central venous access is contraindicated . We retrospectively reviewed 171 patients who received total parenteral nutrition via a central venous triple-lumen catheter and compared the rates of infections in femoral vs nonfemoral access . A literature review was performed to identify associated complications of and appropriate indications for femoral catheter use . In the 171 patients studied, 355 triple-lumen catheters were placed; these included 331 nonfemoral catheters and 24 femoral catheters . Femoral catheters were placed in nine patients . Femoral catheters had a greater incidence of positive tips (42% vs 6.9%, p < .001) and related bacteremia (16.7% vs 1.8%, p = .002) than did nonfemoral catheters . The organisms most commonly isolated from the blood and catheter tips of both catheter access sites were methicillin-resistant Staphylococcus epidermidis and Candida . The use of femoral catheters for central venous access for total parenteral nutrition administration results in an increased risk of infectious complications.

J Bone Joint Surg Am, 1995 Apr, 77(4), 524 - 9
Delayed infections following posterior spinal instrumentation for the treatment of idiopathic scoliosis; Richards BS; Ten patients who had been managed with posterior spinal arthrodesis and Texas Scottish Rite Hospital instrumentation because of idiopathic scoliosis had a delayed deep wound infection at an average of twenty-five months after the operation . The signs of infection included spontaneous drainage in eight patients and fluctuance in two patients . In addition, six patients--including five of the eight who had drainage--had mild pain in the back . The average erythrocyte sedimentation rate was thirty-nine millimeters per hour (range, nineteen to eighty-one millimeters per hour) . The instrumentation was removed from all of the patients . In two patients, a pseudarthrosis that had not been noted on preoperative radiographs was noted intraoperatively; in both patients, the pseudarthrosis occurred at a level at which two hooks had been placed in one intervertebral space . Primary closure was performed in seven patients, and delayed primary closure was performed on the third postoperative day in three patients . All wounds healed uneventfully . Cultures of specimens taken from deep within the wound were positive for Propionibacterium acnes (five patients), Staphylococcus epidermidis (two patients), a rare coagulase-negative Staphylococcus species (one patient), or Micrococcus varians (one patient) . No organisms grew on culture of the specimen obtained from the remaining patient . Propionibacterium acnes required an extended period of incubation before identification . Antibiotics were administered parenterally to all of the patients after the removal of the hardware, and this treatment was followed by oral administration of antibiotics for nine of the patients . We suspect--but can not prove--that several of the delayed infections resulted from intraoperative seeding and remained subclinical for an extended period of time.

Arch Surg, 1995 Apr, 130(4), 446 - 7
Tropical pyomyositis presenting in the upper extremity; Michaels BM et al.; Tropical pyomyositis is a staphylococcal infection, usually of a single large muscle, most commonly seen in young men in tropical regions . The following presents a case of tropical pyomyositis in a 62-year-old man that affected all four extremities, including the muscles of the forearms . Computed tomography was a useful guide for directing surgical explorations . To decrease the degree of disability from the multiple operative sites, we successfully used limited longitudinal incisions and conservative debridements.

Am J Kidney Dis, 1995 Apr, 25(4), 593 - 6
Catheter-related sepsis complicating long-term, tunnelled central venous dialysis catheters: management by guidewire exchange; Shaffer D; Standard therapy of catheter-related sepsis of long-term, tunnelled, silicone dialysis catheters is catheter removal, parenteral antibiotics, and catheter replacement in a new venous site after documented clearing of bacteremia . This leads to loss of future venous access sites . Thirteen consecutive cases of dialysis catheter-related sepsis in 10 patients successfully managed by guidewire exchange with preservation of the same central venous access site are reported . Although the most common cause of catheter sepsis in this series was coagulase-negative staphylococcus, guidewire exchange also was successful in cases due to gram-negative rods and yeast . To preserve future venous access sites in the chronic hemodialysis population, long-term, tunnelled dialysis catheter-related sepsis should be managed by a short course of parenteral antibiotics and by changing the catheter over a guidewire using the same venous insertion site.

Cell Immunol, 1995 Apr 1, 161(2), 188 - 94
Heligmosomoides polygyrus adult worm homogenate superantigen: presentation to T cells requires MHC class I positive accessory cells; Robinson M et al.; A series of experiments were carried out to further characterize the previously discovered Heligmosomoides polygyrus, adult worm homogenate (AWH), superantigen . AWH, in contrast to staphylococcal enterotoxin B (SEB) superantigen, was totally unable to stimulate naive thymocytes, in either the presence or the absence of exogenous accessory cells (AC) . Experiments using AC from B10 congenic mice failed to indicate a requirement for a specific MHC haplotype for successful presentation of the AWH superantigen and also indicated that the presence or absence of the H-2,E molecule on AC did not affect AWH stimulation of T cell hybridomas . Furthermore, AWH was found to require the presence of MHC class I-positive AC in order to stimulate T cell hybridomas, while, in contrast, the absence of MHC Class II on AC did not affect the superantigenic properties of AWH . Initial characterization of the T cell hybridomas stimulated by the AWH superantigen, indicated that all were CD4-positive and that three of them expressed TCR V beta 8.1 . Hence AWH superantigen can stimulate TCR V beta 8.1/CD4-positive T cells only in the presence of MHC Class I-positive AC.

Eur Heart J, 1995 Apr, 16 Suppl B, 94 - 8
Surgical treatment of infective endocarditis; Acar J et al.; Optimal timing of surgery in infective endocarditis (IE) depends mainly on the haemodynamic tolerance of the patient . Emergency surgery is required in cases of refractory heart failure due to valvular lesions, intracardiac fistulas and high grade cardiac conduction abnormalities caused by septal abscesses . Surgery must be considered in all patients who have undergone a transient episode of heart failure such as a pulmonary oedema and it must be early--within 2 or 3 weeks of starting antibiotic therapy in patients with aortic regurgitation . Bacteriological indications are less frequent: persistent sepsis beyond the first week in spite of medical therapy, mycotic IE or prosthetic valve endocarditis caused by Gram-negative or staphylococcal organisms . Some complications may swing the argument in favour of surgery: detection of root abscesses or mycotic aneurysms using transoesophageal echocardiography, and systemic embolisms with persistent, large and mobile vegetative lesions . Mortality rate depends on the haemodynamic status but also on the severity of anatomical lesions, on the type of endocarditis (native or prosthetic valve), on the type of surgery and on bacterial aetiologies . It varies between 5% and 30% . The late postoperative outcome is good . The actuarial survival rate at 8 years was 70% in our series of 31 patients with aortic regurgitation and early surgery . In mitral regurgitation, conservative surgery is possible in most cases . In our department, 48 patients with mitral bacterial lesions have been operated on with conservative surgery without operative mortality . IE was active in 14, recent in 12 and had occurred earlier in 22.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur Heart J, 1995 Apr, 16 Suppl B, 63 - 7
Echocardiographic assessment of prosthetic valve endocarditis; Vered Z et al.; Prosthetic valve endocarditis is still a very serious complication, carrying an incidence of death between 30 and 70% in some series . Therefore early and accurate diagnosis is crucial . Early (less than 60 days post surgery) endocarditis is usually a fulminant disease, where staphylococcal infection is most common . Late prosthetic endocarditis resembles more closely other forms of the disease . Conventional echocardiography is useful in the evaluation of prosthetic valve function, but it is very limited in the demonstration of infective lesions, primarily because of acoustic shadowing . Transoesophageal echocardiography (TE) enables high resolution imaging of the heart without chest wall interference, and viewing of the heart from the posterior (atrial, low pressure) side, where most of the vegetations are expected to be found in both mitral and tricuspid positions . It also enables better visualization of the left ventricular outflow tract, where aortic prosthetic vegetations tend to be present . Furthermore, transoesophageal echocardiography allows accurate diagnosis of some of the common complications of endocarditis: abscess/cavity formation; mycotic aneurysm; prosthetic valve dehiscence and regurgitation . In spite of these advantages, limitations should be recognized . Struts are commonly seen on transoesophageal echocardiography following surgery and should not be confused with vegetations . Similarly, normal prosthetic regurgitation should not be confused with paravalvar leakage . Nevertheless, transoesophageal echocardiography, when expertly used, changes the possibility for early and more accurate diagnosis of prosthetic valve endocarditis dramatically . Transoesophageal echocardiography should be included among the major criteria in the diagnosis and follow-up of prosthetic valve endocarditis.

Eur Heart J, 1995 Apr, 16 Suppl B, 32 - 8
Early infective endocarditis on prosthetic valves; Chastre J et al.; Despite major advances in cardiovascular surgical techniques and routine use of prophylactic antimicrobial agents, prosthetic valve endocarditis (PVE) continues to complicate the course of a small percentage of patients after cardiac valve replacement . Using actuarial methods to describe the risk of PVE after valve implementation, several studies have shown that its incidence peaked at around 5 weeks and levelled off to a stable rate by 12 months, for a cumulative risk of 3% at that time . The microbial aetiology of early PVE is dominated by staphylococcal species, S . epidermis and S . aureus accounting for about 30% and 20% of cases, respectively, even though prophylactic regimens used today are targeted against these microorganisms . In nearly all patients, infection spread behind the site of attachment of the valve prosthesis, resulting in valve ring abscesses and valve dehiscence in 60% of cases . Valve obstruction by vegetations is much more uncommon except in patients with mitral or tricuspid valve endocarditis . The clinical course of early PVE tends to be frequently fulminant, with rapid deterioration of the haemodynamic status due to valvular or annular destruction or persistent bacteraemia . In some cases, however, the classic symptoms of endocarditis may be less noticeable because signs related to an initial source of bacteria, such as sternotomy wound infection, may be more prominent . While various non-invasive and invasive studies have been proposed to aid in the diagnosis of PVE, transoesophageal echocardiography is now the technique of choice for that purpose, as well as for detecting prosthetic valve dysfunction and other intracardiac complications of PVE.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibiot Khimioter, 1995 Apr, 40(4), 37 - 9
{Therapeutic effectiveness of furazolidone and difuracil (PAP-49) in experimental staphylococcal and Escherichia septicemia}; Terekhov VI et al.; The therapeutic efficacy of a new nitrofuran, N-(5-nitrofurilidene)-5-nitrofuran-2-(N'-acetyl)carboxamidra zone (difuracil or PAP-49) in experimental staphylococcal and Coli septicemia was stated and compared to that of furazolidone . It was shown in the models of staphylococcal and Coli septicemia of albino mice that by its therapeutic efficacy difuracil (PAP-49) was 1.3 times more active as furazolidone and by 1-2 days earlier eradicated the pathogens in the host.

Eur J Clin Microbiol Infect Dis, 1995 Apr, 14(4), 359 - 62
Influence of the incubation atmosphere on the production of slime by Staphylococcus epidermidis; Perez-Giraldo C et al.; The influence of various incubation atmospheres on the growth and slime production of 23 Staphylococcus epidermidis strains was studied . The atmospheres evaluated were aerobiosis (control), anaerobiosis, candle jar, 5% CO2 and 10% CO2 . As compared to the aerobic control, growth was 55.7 +/- 19% (p < 0.01) in anaerobic incubation, 113.7 +/- 12% (p < 0.01) in 5% CO2, 112.8 +/- 13% (p < 0.01) in 10% CO2 and 106.4 +/- 7% (p > 0.1) in the candle jar . The slime production in relation to the aerobic control was 20.3 +/- 19% in anaerobiosis (p < 0.01), 22.3 +/- 27% (p < 0.01) in 5% CO2, 29.4 +/- 39% (p < 0.01) in 10% CO2 and 68.3 +/- 26% (p > 0.1) in the candle jar . The results of this study may explain the discrepancies which have been noted on occasion between slime formation data and pathogenicity.

Eur J Clin Microbiol Infect Dis, 1995 Apr, 14(4), 342 - 6
Use of ribotyping to investigate tracheal colonisation by Staphylococcus epidermidis as a source of bacteremia in ventilated newborns; Betremieux P et al.; Ribotyping was used to determine whether a relationship exists between endotracheal tube colonisation with Staphylococcus epidermidis and bacteremia with this organism . Over a three-week period, four mechanically ventilated preterm babies presented with five episodes of infection and bacteremia . For each blood specimen obtained for culture a tracheal aspirate sample was collected at the same time by suctioning . After DNA extraction and cleavage by EcoRI, hybridisation was performed with a digoxigenin-labelled 16S-rDNA probe from Escherichia coli . Five different band patterns were recognised on the membrane . In two children the same band pattern was found in Staphylococcus epidermidis isolated from both blood and tracheal aspirate . Ribotyping thus could be used to differentiate a series of infections from an outbreak and showed that a relationship may exist between tracheal colonisation and bacteremia in mechanically ventilated newborns.

Clin Orthop, 1995 Apr, (313), 214 - 9
Septic sacroiliitis; Osman AA et al.; The authors treated 31 patients with septic arthritis of the sacroiliac joint (14 patients with tuberculosis, 7 acute staphylococcal, 6 gonococcal, and 4 typhoid) . The clinical presentation was vague and nonspecific, but most patients reported buttock pain, low back pain and difficulty walking . In 28 patients, the diagnosis was established after clinical examination, bone scans, hematologic investigations, and blood cultures . The diagnosis was established through arthrocentesis of the sacroiliac joint in 9 patients . In 3 patients, there was a delay in diagnosis . Twenty-nine patients improved on a conservative regimen of bed rest and antibiotics . Two patients required open drainage because of a large buttock abscess that was secondary to tuberculosis sarcoiliitis.

FEMS Immunol Med Microbiol, 1995 Apr, 11(2), 91 - 7
Staphylococcal enterotoxin B toxicity in BALB/c mice: effect on T-cells, plasma cytokine levels and biochemical markers; Wood AC et al.; Groups of BALB/c mice were treated with a sub-lethal dose (60 micrograms) of staphylococcal enterotoxin B (SEB) intraperitoneally and were sacrificed at 2, 5, 8, or 10 h post-injection . Organ, blood plasma and lymph node samples from these mice were analyzed . Plasma levels of urea, creatinine and alanine aminotransferase were significantly raised above normal by 5 h post-injection . However, alkaline phosphatase levels showed an erratic increase after toxin administration and, after administration of 10-40 microgramS SEB per mouse, were consistently at least 30% below normal levels at 24 h post-injection . Weight change was also monitored but found to be inconsistent . Lung, spleen and kidney samples appeared normal on histopathological examination, but liver samples showed minor polymorph infiltration and congestion . TNF-alpha, and IL-1 alpha levels in the plasma were raised by 8 h to picogram levels per ml of plasma, whereas IFN-gamma and IL-2 were raised by 2 h to nanogram levels per ml of plasma . Lymph node cells taken from mice treated with toxin were given a secondary stimulation with toxin in vitro . Although the response of the cells was lower than normal on assay at four days, a time response curve showed a peak in cell responsiveness to secondary stimulation with toxin at three days . These data indicate that biochemical markers and cytokine levels are affected by the administration of SEB to mice and may be used as indicators of toxicity.

Histochem J, 1995 Apr, 27(4), 300 - 8
Binding of bacterial toxins to glycoproteins in the envelopes of rainbow trout eggs; Kudo S et al.; The ability of the vitelline and fertilization envelopes of rainbow trout eggs to trap toxins was investigated using cholera enterotoxin B and staphylococcal enterotoxin B in cytochemical or immunocytochemical experiments . Extracts from both envelopes were investigated by immunoblot analysis to identify toxin-binding proteins after SDS-PAGE . Binding studies of cholera enterotoxin B to vitelline envelopes and fertilization envelopes revealed a greater reactive intensity in the former . Treatment with neuraminidase enhanced the reactive intensity (or deposit) in the vitelline envelope and fertilization envelope outermost layers, with more conspicuous reactivity in the former . Cytochemical experiments showed that exogenous ganglioside GM1 considerably enhanced cholera enterotoxin B binding to vitelline and fertilization envelopes . This enhancement was shown by an intense reactivity following the occurrence of new binding sites on the vitelline envelope inner surface and the inner wall of the zona radiata, a simultaneous extreme reduction in the reactivity of the vitelline envelope outermost layer, and a striking increase in reactive products in the fertilization envelope outermost layer . The surface region of the vitelline or fertilization envelope outermost layer was the binding site for staphylococcal enterotoxin B, and neuraminidase treatment caused a considerable reduction of reactive products in these areas . Immunoblot analysis of cholera enterotoxin B- or staphylococcal enterotoxin B-binding substances in extracts from the vitelline envelopes or fertilization envelopes demonstrated that the great majority of the binding substances are glycoproteins . The present results suggest that glycoproteins constituting the vitelline envelope or fertilization envelope may contribute to the protection of the egg itself or the embryo by trapping noxious toxins.

J Biomed Mater Res, 1995 Apr, 29(4), 485 - 93
Adhesion of Staphylococcus epidermidis to biomedical polymers: contributions of surface thermodynamics and hemodynamic shear conditions; Wang IW et al.; Adhesion studies of Staphylococcus epidermidis RP62A were conducted using a rotating disk system to determine the roles of surface physicochemistry and topographies under physiologic shear conditions . Six materials were investigated: biomedical reference polyethylene and polydimethylsiloxane; argon plasma-treated reference polyethylene (Ar-PE); Silastic; expanded polytetrafluoroethylene; and woven Dacron . All of the polymers except Dacron demonstrated reduced bacterial adhesion with increasing shear stress . Argon plasma treatment of polyethylene reduced the level of staphylococcal adhesion . Adsorption of human plasma proteins effected significantly lower numbers of adherent bacteria . The lowest adhesion was observed for Ar-PE in 1% human plasma protein solution, whereas Dacron had the highest number of adherent bacteria . The high adhesion on Dacron was attributed to increased bacterial flux caused by topography-induced turbulent flow and physical entrapment of the bacteria in the fiber interstices . The results indicate that the driving force for S . epidermidis adhesion is strongly influenced by substrate physicochemistry, but this may be dominated by physical forces such as shear and turbulence.

J Biomed Mater Res, 1995 Apr, 29(4), 455 - 66
Bacteria/blood/material interactions . I . Injected and preseeded slime-forming Staphylococcus epidermidis in flowing blood with biomaterials; Brunstedt MR et al.; Blood-material interactions were studied using in vitro recirculation with human blood, slime-forming Staphylococcus epidermidis, and cardiovascular materials . Staphylococcus epidermidis, under preseeded or injected conditions, adhered to nonsmooth materials and elevated plasma levels of fibrinopeptide A (FpA) and C3a in the presence of all materials . Increased white blood cell (WBC) and platelet adhesion and thrombospondin and platelet factor 4 (PF4) release were noted for respective materials in the presence of injected bacteria . Materials that adhered significant quantities of injected S . epidermidis exhibited low levels of adsorbed proteins . Materials with high levels of preseeded S . epidermidis showed high levels of adsorbed proteins . Adhesion of preseeded bacteria and blood plasma elevations of C3a and FpA were lowest on semicrystalline polymer substrates, intermediate on halogenated substrates, and highest on amorphous substrates . In the presence of injected bacteria, WBCs and platelets adhered at earlier recirculation times to amorphous substrates than to semicrystalline substrates.

Protein Sci, 1995 Apr, 4(4), 636 - 54
Proline cis-trans isomerization in staphylococcal nuclease: multi-substrate free energy perturbation calculations; Hodel A et al.; Staphylococcal nuclease A exists in two folded forms that differ in the isomerization state of the Lys 116-Pro 117 peptide bond . The dominant form (90% occupancy) adopts a cis peptide bond, which is observed in the crystal structure . NMR studies show that the relatively small difference in free energy between the cis and trans forms (delta Gcis-->trans approximately 1.2 kcal/mol) results from large and nearly compensating differences in enthalpy and entropy (delta Hcis-->trans approximately delta TScis-->trans approximately 10 kcal/mol) . There is evidence from X-ray crystal structures that the structural differences between the cis and the trans forms of nuclease are confined to the conformation of residues 112-117, a solvated protein loop . Here, we obtain a thermodynamic and structural description of the conformational equilibrium of this protein loop through an exhaustive conformational search that identified several substates followed by free energy simulations between the substrates . By partitioning the search into conformational substates, we overcame the multiple minima problem in this particular case and obtained precise and reproducible free energy values . The protein and water environment was implicitly modeled by appropriately chosen nonbonded terms between the explicitly treated loop and the rest of the protein . These simulations correctly predicted a small free energy difference between the cis and trans forms composed of larger, compensating differences in enthalpy and entropy . The structural predictions of these simulations were qualitatively consistent with known X-ray structures of nuclease variants and yield a model of the unknown minor trans conformation.

Eur J Pediatr, 1995 Apr, 154(4), 295 - 8
Recombinant human interferon-gamma in patients with chronic granulomatous disease--European follow up study; Weening RS et al.; This was an uncontrolled, open-label follow up study of a previous 12-month, randomized, double-blind, placebo-controlled trial performed to assess the long-term efficacy and safety of Recombinant Human Interferon Gamma (rIFN-gamma) in patients with chronic granulomatous disease (CGD) . In two centres, 28 patients (24 male, 4 female) with a mean age of 16 years (range 3-37) entered the open-label phase . The patients were treated for a mean of 880 days (range 97-1375 days) . Visits were scheduled every 180 days and patients completed one to six visits . rIFN-gamma was administered subcutaneously three times weekly at a dose of 0.05 mg per m2 . During the open-label phase of the study 12 patients experienced a serious infection requiring hospitalization within 880 days . The median infection-free time was 993 days . No obvious increase of infections over time was seen . Phagocyte superoxide anion production and phagocyte staphylococcal killing were not influenced by therapy . Seven patients were withdrawn from the study, one because of an adverse reaction, three on their own wish and the other three because they changed to another trial . No patient died during the study . Conclusion . Treatment of patients with CGD with intracellular active antibiotics and additional interferon gamma as infection prophylaxis is safe and justified.

Pediatr Res, 1995 Apr, 37(4 Pt 1), 455 - 9
Frequency and cytokine phenotype of blood T cells from premature infants responding to staphylococcal enterotoxin B; Hayward AR et al.; The responder cell frequency (RCF) of premature (< 1900 g birth weight) infants' blood lymphocytes, which proliferate in cultures stimulated by staphylococcal enterotoxin B, falls from 1:3400 to about 1:8000 during the first 2 wk of life . Term infants, in contrast, show no fall in RCF . The reduced RCF in the premature infants affected cells that make interferon-gamma more than cells making IL-4 . The reduced RCF was accompanied by a fall in the fraction of V beta 3+ T cells that entered cell cycle in stimulated cultures . The RCF of premature infants' T cells was increased in cultures supplemented with irradiated monocytes from adults . Addition of IL-4 (but not IL-2, IL-6, or indomethacin) increased the RCF and fraction of cells entering cell cycle of the premature infants . The data suggest that postnatal environmental factors limit the ability of premature infants' monocytes to support a T-cell response to staphylococcal enterotoxin B in vitro and that this limitation is overcome by adding IL-4.

Diagn Microbiol Infect Dis, 1995 Apr, 21(4), 227 - 30
Reevaluations of disk diffusion susceptibility testing interpretive criteria for lomefloxacin and norfloxacin using fluoroquinolone-resistant isolates; Cormican MG et al.; The current National Committee for Clinical Laboratory Standards interpretive criteria for disk diffusion susceptibility testing of lomefloxacin and norfloxacin were reevaluated using a test panel of 298 bacteria (200 with ciprofloxacin minimum inhibitory concentrations (MICs) at > or = 4 micrograms/ml) . MICs were correlated with the diameter of zones of inhibition by regression statistics and error-rate bounding methods . Modifications of the interpretive criteria for lomefloxacin disk susceptibility testing are proposed as follows: susceptible at > or = 20 mm and resistant at < or = 16 mm, a 2-mm decrease of existing break-point zones . These criteria result in an absolute interpretive agreement of 95.3% with a very major (false susceptible) error rate of only 0.7% . The currently used criteria for norfloxacin (susceptible at > or = 16 mm and resistant at < or = 13 mm) were validated, and these break-points had an absolute interpretive correlation between methods of 91.9% . The change proposed for lomefloxacin disk interpretations would minimize minor and major errors most often reported for Staphylococcus saprophyticus isolates.

Cell Immunol, 1995 Apr 1, 161(2), 158 - 65
Superantigens induce primary T cell responses to soluble autoantigens by a non-V beta-specific mechanism of bystander activation; Rott O et al.; Superantigens have been suggested to act as powerful TCR V beta-specific inducers of T cell reactivity in autoimmune diseases . We have investigated the capacity of staphylococcal enterotoxins (SE) to prime autoreactive T cell responses in naive animals in the Lewis rat model of experimental autoimmune encephalomyelitis (EAE), where myelin basic protein (MBP)-specific CD4+ effector T cells express almost exclusively V beta 8.2 TCR elements . By taking advantage of the reactivity of V beta 8.2+ MBP-specific T cells to SEE but not to other SEs in vitro, we estimated the potential of different SEs (SEA, SEB, and SEE) to induce a primary T cell response to soluble MBP in vivo . Upon immunization of naive rats with soluble MBP alone or MBP and SEB (which is only a very weak superantigen for rat T cells), no MBP-responses could be retrieved . Similarly, when coimmunizing naive rats with MBP and V beta 8.2-activating SEE, no autoreactivity was inducible . By contrast, coimmunization of animals with soluble MBP and the superantigen SEA that is strongly activating various T cell subpopulations in Lewis rats but not V beta 8.2+ (i.e., potentially MBP reactive) T cells led to a significant primary MBP-specific T cell autoreactivity . These SEA-induced MBP-reactive T cells expressed V beta 8.2 TCRs at levels similar to those seen in autoreactive T cells conventionally induced by immunization with MBP administered in complete Freund's adjuvant (CFA) and could induce disease in a transfer model of EAE . Thus, our results are consistent with the notion that superantigens are able to induce primary T cell responses to soluble autoantigens by a non-V beta specific mechanism of bystander priming.

J Immunol, 1995 Apr 1, 154(7), 3204 - 12
Unique cytokine production profile of anergic human T cells in SCID-hu mice after staphylococcal enterotoxin B administration; Schols D et al.; Severe combined immunodeficient mice transplanted with human organs (SCID-hu mice), provide a unique in vivo model for studying human intrathymic T cell selection and development of tolerance . In vivo administration of staphylococcal enterotoxin B (SEB) to SCID-hu mice causes intrathymic clonal deletion of SEB-specific V beta+ T cells that occurs already at the immature CD4+8+ double positive stage . The expression of activation markers such as CD25, CD71, and HLA-DR was specifically increased on V beta+ T cells responding to SEB . The remaining SEB-specific human T cells that had not been deleted in vivo failed to proliferate when rechallenged with SEB in vitro . These SEB-specific T cells that were rendered anergic in vivo had a unique cytokine production profile . They failed to produce IL-2, which correlated with the lack of proliferation of these cells . In addition, they failed to produce TNF-alpha . However, the anergized T cells synthesized considerable amounts of IFN-gamma, granulocyte-macrophage CSF and IL-10 after SEB stimulation . This clonal anergy can be completely reversed in vitro by stimulating the SEB-specific cells in the presence of exogenous IL-2 or by triggering of the CD28/CTLA-4 activation pathway . Under these stimulation conditions, anergic T cells produced levels of IL-2 and TNF-alpha that were comparable to their non-anergized counterparts, whereas the levels of granulocyte-macrophage CSF, IL-10 and IFN-gamma production were even higher . Collectively, these data demonstrate that in vivo administration of SEB to SCID-hu mice leads to activation, deletion, and anergy of SEB-specific human thymocytes and that the production of IL-2 and TNF-alpha is selectively switched off in these anergic T cells.

Riv Inferm, 1995 Apr-Jun, 14(2), 62 - 6
{Overview of nosocomial infections in a neonatal intensive care unit (1989-1993)}; Cologna M; Newborns admitted to any Intensive Care Unit (ICU) are more at risk for nosocomial infections . The author analyzed the incidence of nosocomial bacteriaemias occurred in the neonatal ICU of Trento, especially focusing on the relationship between weight at delivery, incidence of infections and mortality . The incidence of bacteriaemiae during the observation period (1.189-31.12.93) accounted for 3.5%, and 2.8% were classified as nosocomial . The most frequently involved organism was the Staphylococcus PCN (62% of bacteriaemiae) . High risk newborns weighted < 1500 g; 10.7% were affected by nosocomial bactaeriaemia, representing the 56.1% of all the observed bacteriaemiae . A strong association (p < 0.05) was also observed between bactaeriaemiae and mortality, in newborns weighting less than 1000 g . Since low and very low birth weigh infants are particularly at risk for infections, special attention should be paid to the invasive techniques and a systematic reporting and documentation of epidemiology of infections, in order to monitor impact of modifications in caring techniques is warranted.

FEBS Lett, 1995 Mar 27, 362(1), 29 - 33
A cell-free protein translocation system prepared entirely from a gram-positive organism; Schimz KL et al.; A cell-free protein translocation system derived exclusively from a Gram-positive bacterium is described here for the first time . Highly efficient in vitro synthesis of plasmid encoded preprolipase of Staphylococcus hyicus is accomplished by coupled transcription/translation using either a cytosolic extract of S . carnosus alone or in combination with T7-RNA-polymerase . Addition of inside-out cytoplasmic membrane vesicles of S . carnosus leads to the partial conversion (processing) of preprolipase to prolipase . In addition, as shown in a protease protection assay, a significant part of preprolipase plus prolipase is translocated in vitro into the lumen of the vesicles . Translocation of preprolipase into the membrane vesicles requires the proton-motive force and the S . carnosus SecA protein.

J Immunol, 1995 Mar 15, 154(6), 2600 - 11
Selective expansion followed by profound deletion of mature V beta 8.3+ T cells in vivo after exposure to the superantigenic lectin Urtica dioica agglutinin; Galelli A et al.; Urtica dioica agglutinin (UDA) is a superantigen that, in vitro, binds to specific carbohydrate structures on class II and induces a sixfold enrichment of V beta 8.3+ BALB/c mice splenic T cells . Superantigens have pleiotropic effects in vivo, causing the activation, proliferation, and deletion of specific T cells, but are heterogenous in regard to their effects on T cell tolerization . We, therefore, compared the responses of peripheral T cells from adult BALB/c mice with the i.v . injection of 50 micrograms UDA or the bacterial superantigen staphylococcal enterotoxin B (SEB) that also recognizes the V beta 8.3 gene product . The data presented indicate that activation, clonal expansion, anergy, and death of V beta 8.3+ T cells occur sequentially after UDA administration . Two days after UDA injection, the proportion of V beta 8.3+ T cells in the periphery is elevated to approximately twice that of normal mice . This expansion occurs in both CD4+ and CD8+ subsets . V beta 8.3+ T cells from UDA-primed mice are anergic to UDA restimulation and fail to proliferate or to produce IL-2 . Futhermore, the proliferation of V beta 8.3+ T cells is followed by their rapid disappearance concomitant with their specific elimination by apoptosis . In 1 wk, all CD4+ V beta 8.3+ peripheral T cells are deleted . The decline of V beta 8.3+ T cells in the CD4+ subset is more than in the CD8+ subset . This occurs in thymectomized and in thymus-intact animals . Two months after UDA priming, the percentage of V beta 8.3+ T cells is still lower than in control mice.

J Med Microbiol, 1995 Mar, 42(3), 156 - 60
Chronic bacteraemia due to Staphylococcus epidermidis after bone marrow transplantation; Lina B et al.; A chronic bacteraemia due to Staphylococcus epidermidis occurred in a patient undergoing allogeneic bone marrow transplantation . Forty-two S . epidermidis isolates were obtained from blood cultures over a period of 5 months . Isolates were separated into three groups by SmaI macrorestriction characterisation with pulsed-field gel electrophoresis (PFE-1, one isolate; PFE-2, 32 isolates; PFE-3, nine isolates) . Differences were detected in antimicrobial susceptibility patterns among isolates belonging to group PFE-2 . The two strains, PFE-2 and PFE-3, were both responsible for the chronic bacteraemia and were isolated during different admissions to the hospital . A central venous catheter was the portal of entry for PFE-2 . DNA macro-restriction analysis with pulsed-field gel electrophoresis was helpful in the epidemiological investigation of this S . epidermidis chronic bacteraemia.

J Leukoc Biol, 1995 Mar, 57(3), 450 - 4
Differential inhibitory effects of interleukin-10, interleukin-4, and dexamethasone on staphylococcal enterotoxin-induced cytokine production and T cell activation; Krakauer T; The cytokine profile of human peripheral blood mononuclear cells (PBMC) stimulated by staphylococcal enterotoxin (SE) A and B was examined . Production of tumor necrosis factor (TNF alpha), interleukin (IL)-1, IL-6, IL-2, and gamma interferon (IFN-gamma) was observed . In contrast, Th2 cytokines IL-4 and IL-10 were absent from SEA- or SEB-stimulated PBMC . Moreover, adding IL-10 to SE-stimulated PBMC inhibited the production of IL-1, IL-6, TNF alpha, and IFN gamma by 50 to 80% but had less effect (8-30%) on T cell proliferation . IL-4 was less effective than IL-10 in inhibiting cytokine production and enhanced T cell proliferation by SEA or SEB . The anti-inflammatory agent, dexamethasone, was the most potent agent in controlling the SE-mediated effects as evidenced by inhibited T cell proliferation (55%) and reduced levels of IL-1, IL-6, and IFN gamma (60% to 100%) and TNF alpha (50%) . Reducing levels of toxic mediators such as TNF alpha, IL-1, IL-6, and IFN gamma by dexamethasone in SE-induced T cell responses may be a useful therapeutic strategy to circumvent SE toxicity and pathogenesis.

Clin Exp Immunol, 1995 Mar, 99(3), 398 - 403
Cytotoxic activity of V beta 8+ T cells in Crohn's disease: the role of bacterial superantigens; Baca-Estrada ME et al.; In Crohn's disease, disease-related stimuli could alter the T cell receptor (TCR) repertoire . To examine the possibility that changes in function may occur in T cell subsets without obvious changes in expression of TCR, we analysed the TCR repertoire of cytotoxic T lymphocytes in Crohn's disease peripheral blood . Furthermore, we examined the effect of bacterial superantigens, staphylococcal enterotoxin B (SEB) and E (SEE) on the cytotoxic function of T cell subsets bearing different TCR V genes using MoAbs specific for CD3 and TCR V gene products in a redirected cytotoxicity assay . There was no difference between patients and controls in the cytotoxicity measured in concanavalin A (Con A)-stimulated peripheral blood mononuclear cells (PBMC) with anti-CD3 or with six of seven anti-TCR V gene MoAbs . However, the cytotoxicity of V beta 8 T cells was decreased in Crohn's disease patients . This was not due to a decrease in total or CD8+ T cells expressing V beta 8 . Furthermore, in normal subjects, PBMC stimulation with SEE and SEB selectively expanded and increased the cytotoxicity of V beta 8 and V beta 12 T cells, respectively . In Crohn's disease, although SEB stimulation increased the number and cytolytic function of the V beta 12 subset, SEE stimulation failed to increase cytolytic activity of V beta 8+ T cells in spite of the expansion of V beta 8+ T cells . These results suggest that the changes in cytotoxic function observed in V beta 8 T cells in Crohn's patients may reflect previous exposure to a V beta 8-selective superantigen.

Clin Exp Immunol, 1995 Mar, 99(3), 345 - 51
Synovial cells are potent antigen-presenting cells for superantigen, staphylococcal enterotoxin B (SEB); Origuchi T et al.; There is ample evidence suggesting that superantigens may act as a triggering factor in the pathogenesis of rheumatoid arthritis (RA) . We investigated whether superantigen could activate T cells in the presence of synovial cells . T cells were cultured with SEB in the presence of interferon-gamma (IFN-gamma)-treated synovial cells . T cell proliferation and activation were assessed by 3H-thymidine incorporation and IL-2 production . The expression of HLA class II antigens and adhesion molecules on synovial cells was detected by flow cytometer . In the presence of IFN-gamma-treated synovial cells, T cells proliferated vigorously and produced IL-2 in response to SEB . A low SEB-induced T cell response was noticed in the presence of untreated synovial cells . Allogeneic as well as autologous IFN-gamma-treated synovial cells markedly enhanced SEB-induced T cell proliferation . IFN-gamma-treated synovial cells had increased expression of HLA class II antigens and intercellular adhesion molecule-1 (ICAM-1) adhesion molecules . MoAbs towards these antigens markedly inhibited the SEB-induced T cell response . These results indicate that activated synovial cells are potent antigen-presenting cells for SEB to T cells, and that superantigens may play a critical role in the pathogenesis of RA through activated synovial cells.

J Thorac Cardiovasc Surg, 1995 Mar, 109(3), 561 - 4
Effect of vancomycin infusion on cardiac function in patients scheduled for cardiac operation; Rosenberg JM et al.; Patients scheduled for cardiac operation often receive vancomycin before the operation to decrease postoperative staphylococcal wound infections . In animal studies, vancomycin depressed cardiac function approximately 15% . Because of the potentially serious consequences of myocardial depression in patients undergoing cardiac operation, we examined the effect of vancomycin infusion on cardiac hemodynamics in patients scheduled for cardiac operation . Patients who were scheduled for cardiac operation and vancomycin prophylaxis were enrolled in our study . After baseline cardiac output, mean arterial pressure, central venous pressure, and pulmonary capillary wedge pressure were measured, 1 gm of vancomycin HCl was infused over 1 hour . Cardiac output, mean arterial pressure, central venous pressure, and pulmonary capillary wedge pressure were measured at 15, 30, 60, 90, and 120 minutes after the start of the infusion . In the 46 patients that completed the study, no significant change was observed in cardiac output or systemic vascular resistance at any time when compared with baseline . Mean arterial pressure increased significantly (p = 0.03) between baseline (90.8 +/- 2.4 standard error of mean) and 90 minutes (94.1 +/- 2.4 standard error of mean) . One patient had a transient 30% fall in mean arterial pressure and systemic vascular resistance with facial flushing during the infusion . In conclusion, we found that vancomycin infusion over 1 hour in patients before cardiac operation is safe and not associated with cardiac depression.

Cell Immunol, 1995 Mar, 161(1), 28 - 33
Patterns of interferon-gamma mRNA expression in toxic shock syndrome toxin-1 expanded V beta 11+ T lymphocytes; Zhao YX et al.; We have recently demonstrated that toxic shock syndrome toxin-1 (TSST-1), a staphylococcal exotoxin with superantigenic properties, is involved in the pathogenesis of septic arthritis . The aim of the current study was to characterize in detail the in vitro properties of TSST-1 with regard to T cell activation patterns . We demonstrate that TSST-1 preferentially expands murine V beta 11+ T lymphocytes and this expansion occurs equally well within CD4 and CD8 compartments . To analyze the functional properties of V beta 11+ T cells expanded in response to TSST-1, we have utilized a combined in situ hybridization and immunocytochemistry method . With this technique we show that TSST-1-stimulated V beta 11+ T cells are an important source of IFN gamma mRNA synthesis, suggesting that this cytokine may participate in superantigen-mediated septic arthritis.

J Urol, 1995 Mar, 153(3 Pt 1), 659 - 61
Inflatable penile implant infection: predisposing factors and treatment suggestions; Wilson SK et al.; A retrospective review of 1,337 consecutive inflatable prosthesis implantations was done to detect predisposing factors for infection . Operations were performed by the same team during a 7-year period . Of the procedures 823 were primary implantations, while the remaining 514 were either revisions (mechanical failure, iatrogenic causes or patient dissatisfaction), salvage operations or removals for infection . The etiology of impotence as a predisposition for infection in primary implantations was significant for spinal cord injury (9% of the cases) and steroid use (50%) . Diabetes had a statistically insignificant 3% risk of infection, with all other causes having a 1% infection rate . A total of 428 revisions was performed with an overall infection rate of 10% . Rates of infection ranged from 8% in nondiabetics to 18% in diabetics requiring revision . Staphylococcus species were the most commonly cultured organisms . Salvage operations (29 cases) of several types were attempted for infected prostheses . Our results with salvage were poor and we currently routinely remove the entire prosthesis for infection except in cases of urethral erosion of 1 cylinder.

J Neurochem, 1995 Mar, 64(3), 1305 - 11
Monoclonal antibodies to the human gamma 2 subunit of the GABAA/benzodiazepine receptors; Fernando LP et al.; The large intracellular loop (IL) of the gamma 2 subunit of the cloned human gamma-aminobutyric acidA (GABAA) receptor (gamma 2 IL) was expressed in bacteria as glutathione-S-transferase and staphylococcal protein A fusion proteins . Mice were immunized with the fusion proteins (one protein per animal), and monoclonal antibodies were obtained . Six monoclonal antibodies reacted with the gamma 2 IL moiety of the fusion proteins . Three of these monoclonal antibodies also immunoprecipitated a high proportion of the GABAA/benzodiazepine receptors from bovine and rat brain and reacted with a wide 44,000-49,000-M(r) peptide band in immunoblots of affinity-purified GABAA receptors . These monoclonal antibodies are valuable reagents for the molecular characterization of the GABAA receptors in various brain regions.

Protein Sci, 1995 Mar, 4(3), 484 - 95
Stabilization of a strained protein loop conformation through protein engineering; Hodel A et al.; Staphylococcal nuclease is found in two folded conformations that differ in the isomerization of the Lys 116-Pro 117 peptide bond, resulting in two different conformations of the residue 112-117 loop . The cis form is favored over the trans with an occupancy of 90% . Previous mutagenesis studies have shown that when Lys 116 is replaced by glycine, a trans conformation is stabilized relative to the cis conformation by the release of steric strain in the trans form . However, when Lys 116 is replaced with alanine, the resulting variant protein is identical to the wild-type protein in its structure and in the dominance of the cis configuration . The results of these studies suggested that any nuclease variant with a non-glycine residue at position 116 should also favor the cis form because of steric requirements of the beta-carbon at this position . In this report, we present a structural analysis of four nuclease variants with substitutions at position 116 . Two variants, K116E and K116M, follow the "beta-carbon" hypothesis by favoring the cis form . Furthermore, the crystal structure of K116E is nearly identical to that of the wild-type protein . Two additional variants, K116D and K116N, provide exceptions to this simple "beta-carbon" rule in that the trans conformation is stabilized relative to the cis configuration by these substitutions . Crystallographic data indicate that this stabilization is effected through the addition of tertiary interactions between the side chain of position 116 with the surrounding protein and water structure . The detailed trans conformation of the K116D variant appears to be similar to the trans conformation observed in the K116G variant, suggesting that these two mutations stabilize the same conformation but through different mechanisms.

Int Immunol, 1995 Mar, 7(3), 425 - 34
Selective activation of VH3A10+ rheumatoid factor producing B cells by staphylococcal enterotoxin D; Xie C et al.; Staphylococcal enterotoxin D (SED) is a T cell superantigen which selectively targets alpha beta TCRs bearing particular V beta elements . A second function of SED relates to the preferential activation of a B cell subset characterized by a high frequency of rheumatoid factor (RF) producing B cells . To define the molecular basis of the SED-induced B cell repertoire shift, we have analyzed Ig heavy chain genes in B cell clones expanded after SED stimulation and compared them with B cell clones established in the presence of anti-CD3 stimulated helper cells . Gene segments of the VH3 family were most frequently utilized under both stimulation conditions (42% anti-CD3; 47% SED) . Sequence analysis of VH3 gene segments demonstrated that the repertoire of VH3 elements in B cell clones from SED driven and anti-CD3 driven cultures were distinct (P = 0.01) . RF activity was closely associated with the expression of selected VH3 elements . B cell clones stimulated with SED preferentially expressed VH3A10, whereas VH26 was the gene segment dominantly used in B cell clones expanded with anti-CD3 stimulated helper cells . The usage of JH and DH elements was indistinguishable in SED and anti-CD3 driven B cell clones, suggesting that SED targets VH3+ B cells through a VH-specific mechanism . Comparison of the closely related sequences of the SED responsive VH3A10 and the SED non-responsive VH26 element suggested a role of a sequence polymorphism in the CDR2 reminiscent of B cell reactivity to conventional antigens . In contrast to conventional antigens, SED can induce differentiation of a high frequency of naive B cells . Thus, this staphylococcal enterotoxin combines selective activation of T cells with selective activation of B cells and might be able to direct T cell help to RF producing B cells.

Lett Appl Microbiol, 1995 Mar, 20(3), 180 - 3
Insertion elements in Staphylococcus intermedius; Hesselbarth J et al.; Staphylococcus intermedius cultures from dogs, pigeons, horses and mink were investigated for the prevalence of the insertion elements IS256 and IS257 in relation to their antibiotic resistance . Copies of IS256 could not be detected in any of the Staph . intermedius isolates tested whereas single copies of IS257 occurred in the isolates from dogs and horses . The mink strains did not harbour IS257 elements, whereas Staph . intermedius isolates from pigeons carried multiple copies of IS257 as predicted from the hybridization patterns obtained with a gene probe derived from the internal part of the IS257-encoded transposase gene . Independently of the origin of the Staph . intermedius isolates, all IS257 copies were found to be located in the chromosomal DNA . The large number of chromosomal IS257 copies in the pigeon strains might help to explain chromosomal multiresistance in many of those strains.

Lett Appl Microbiol, 1995 Mar, 20(3), 168 - 70
In vitro indices of tissue adherence in Staphylococcus intermedius; Cree RG et al.; In vitro indices of adherence showed that strains of Staphylococcus intermedius from lesions of canine pyoderma differed from strains isolated from normal carrier sites in that a significantly greater proportion of pyoderma strains adhered to extracellular matrix proteins whilst fewer adhered to polystyrene . Slime production and a hydrophobicity index did not differ between the groups . This suggests that exposure of extracellular matrix proteins due to underlying disease may result in the selection of a narrower spectrum of strains from amongst those at carrier sites.

Chemotherapy, 1995 Mar-Apr, 41(2), 113 - 20
Antibiotic killing of bacteria: comparison of bacteria on surfaces and in liquid, growing and nongrowing; Eng RH et al.; The minimum inhibitory concentrations of antibiotics for bacterial pathogens are derived from broth suspensions (broth dilution) and from nutrient surfaces (agar dilution) . These concentrations may not apply when bacteria are on a nonnutrient surface such as in a foreign body infection . We compared bacteria (Staphylococcus epidermidis and Escherichia coli) broth suspension MBCs with MBCs of the same bacteria when on a nonnutrient surface in broth the growing and nongrowing phases . Bacteria growing on cotton surfaces were much less susceptible to antibiotic killing than when freely suspended in the liquid nutrient . These results alone, independent of host factors, would explain the failure of antibiotics to eradicate infections involving bacteria on foreign body surfaces . The resistance of bacteria to antibiotic killing is not caused by a lack of antibiotic penetration to the site of the bacteria, but by an altered state of the bacteria when they are associated with a surface.

Epidemiol Mikrobiol Imunol, 1995 Mar, 44(1), 36 - 43
{Peroral vaccine therapy in dermatology}; Koukalova D et al.; In a group of 691 patients with different dermatoses the therapeutic effect of two forms of lyophilized vaccines administered orally according to a desensitization (drops) and immunization (tablets) plan . Their effect was as a rule marked already during the third week of treatment . During the final evaluation no signs of the disease were present or great improvement was recorded in 89-100% of the patients suffering from acne, chronic staphylococcal pyoderma or microbial eczema and 62% (drops) or 65% (tablets) of patients with atopic eczema . During treatment the authors did not observe with rare exceptions of focal reactions any undesirable effects . No significant differences were found between the two forms of the drugs as regards therapeutic effects . Contrary to antibiotics, which can cause remission of dermatosis, while early relapses cannot be prevented, oral vaccine treatment leads to long-term (extending over several years) favourable effects on the course of the disease . The immunization pattern used in tablet form vaccines is simpler, packing and storage is simpler, dosage is more accurate and more convenient for the patient.

Immunology, 1995 Mar, 84(3), 360 - 6
Genetic restriction and specificity of the immune response in mice to fusion proteins containing repeated sequences of the Plasmodium falciparum antigen Pf155/RESA; Sjolander A et al.; The genetic restriction and specificity of the immune response in mice to two fusion proteins, ZZ-M3 and ZZ-M5, were studied . These proteins contain two IgG-binding domains (ZZ) from staphylococcal protein A, and repeated sequences from the C-terminal {(VEHDAEEN)5 (VEEN)10} (M3) or central {(VEEPTVADDEH)3(VEEPTVAEEH)2} (M5) regions of the Plasmodium falciparum malaria blood stage antigen Pf155/RESA . Strong antibody and T-cell responses to M3 and M5 were linked to expression of the I-Ak allele, and T-cell responses to the bacterial fusion partner ZZ were restricted to mice of the H-2k haplotype . The response to M5 was less restricted than that to M3, giving intermediate responses in mice of H-2d haplotypes as well . However, ZZ-M5-primed lymph node (LN) cells from these mice were primarily induced to proliferate in vitro by the complete ZZ-M5 construct and not by synthetic peptides representing the repeated subunits in M5 . The reactivity with intact Pf155/RESA in erythrocyte membrane immunofluorescence was weak of antisera from mice immunized with ZZ-M5, whereas the reactivity of antisera from mice immunized with ZZ-M3 roughly paralleled their reactivity with M3 in an enzyme-linked immunosorbent assay (ELISA) . The antibody responses induced by immunization with ZZ-M3 or ZZ-M5 were specific for M3 or M5, respectively, while activated T cells displayed cross-reactivity between M3 and M5 in an in vitro proliferation assay . The results indicate that the assembly of repeated sequences in fusion proteins affects both the MHC class II restriction and the specificity of the induced antibody and T-cell responses.

Br J Dermatol, 1995 Mar, 132(3), 468 - 71
Staphylococcal scalded skin syndrome in an adult . Influence of immune and renal factors; Hardwick N et al.; We report a case of staphylococcal scalded skin syndrome in a 77-year-old man with an infected surgical wound . The patient was immunocompetent and had only mildly impaired renal function . The pathogenic and aetiological factors of the condition are discussed.

Nippon Sanka Fujinka Gakkai Zasshi, 1995 Mar, 47(3), 231 - 6
{Infection route of MRSA to pregnant women and to newborns}; Mitao M et al.; It has been reported that newborn babies are compromised hosts, and to be MRSA carriers means that the danger of occurrence of MRSA infection is high . Our department has focused on improving the hospital environment to guard against hospital infections . Infection from the hospital environment was prevented by looking for staphylococcus bacteria in the nasal cavity during pregnancy, and isolating pregnant MRSA carriers from others . Such effects as MRSA infection of the body members and organs, the newborn babies, nasal cavities of their family members, and discharging MRSA into hospital environments were investigated . 1 . The rate of MRSA carriers among pregnant women was 0.8% (3.7% of pregnant women engaged in medical services, and 0.5% of pregnant women engaged in non-medical services) . In the former case it was suspected that their children became MRSA carriers for some reason and they became infected through their children . In the latter case, there are two possibilities; one was that MRSA was picked up at the place of work was not removed by sterilization, so that they became carriers and the other is infection through children similarly to non-medical workers . 2 . A healthy carrier was observed to be spreading MRSA in to the environment . 3 . In the environment where hospital infections were fully dealt with, MRSA infection of newborn babies was always from puerperal women who were carriers . 4 . The rates for both MRSA carrying in the nasal cavities of newborn babies at the time of dehospitalization and 1-month-old babies were distinctly low, when the nasal MRSA of puerperal was removed by sterilization, compared to without sterilization.(ABSTRACT TRUNCATED AT 250 WORDS)

Zh Mikrobiol Epidemiol Immunobiol, 1995 Mar-Apr, (2), 59 - 63
{The immunomodulating properties of the erythrocytes in an experimental staphylococcal infection}; Barsukov VS et al.; Experiments on 130 mice of different strains have revealed that washed mouse erythrocytes taken on day 7 of experimental staphylococcal infection induce the pronounced increase of humoral immune response to sheep red blood cells in syngeneic recipients, while mouse erythrocytes taken on day 14 of this infection induce immunosuppression . Erythrocytes acquire immunosuppressive properties at the period of red blood regeneration.

Tsitol Genet, 1995 Mar-Apr, 29(2), 45 - 9
{The reaction of anterior hypothalamic nuclei and of the endocrine glands to the administration of staphylococcal peptidoglycan}; Danilova OV et al.; The results of investigation of the influence of Staphylococcus peptidoglycan on supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei, adrenal and thyroid glands of CBA mice was studied . Alterations in the SON and PVN secretory cells were found . The percentage of neurocytes that actively synthesize and secrete the Gomori {correction of Homory}-positive substance increased . Biosynthetic functions and secretion of the hormone was enhanced . In adrenal glands, alterations were found only in the bundled zone.

J Struct Biol, 1995 Mar-Apr, 114(2), 153 - 5
Crystallization and preliminary X-ray analysis of a lipase from Staphylococcus hyicus; Ransac S et al.; Single cyrstals of a lipase from Staphylococcus hyicus have been obtained using a combination of 18 to 24% dimethylsulfoxide and 10% isopropanol as a precipitant . The crystals grow at 4 degrees C in 2-3 months . They belong to the orthorhombic space group P212121 with a = 73.31 A, b = 77.96 A, and c = 169.81 A, with two protein molecules per asymmetrical unit . The crystals diffract to at least 2.8 A resolution and are suitable for an X-ray structure analysis.

Ann Pharmacother, 1995 Mar, 29(3), 267 - 8
Pruritus associated with intravenous rifampin; Walker-Renard P; OBJECTIVE: To report a probable case of moderate pruritus associated with intravenous rifampin . CASE SUMMARY: A 57-year-old woman was started on vancomycin 1 g i.v . q72h following a positive blood culture for methicillin-resistant Staphylococcus epidermidis . The patient's past medical history was significant for uterine carcinoma, radiation-induced enteritis, congestive heart failure, and renal insufficiency . Other therapy consisted of total parenteral nutrition, digoxin 0.125 mg/d i.v., and furosemide 60 mg i.v . prn for edema . Seven days later, the patient's white blood cell count increased and rifampin 600 mg/d i.v . was added to the drug regimen for potential synergy . On day 3 of rifampin therapy, the patient experienced pruritus beginning 30 minutes after the rifampin infusion ended and lasting for 6-8 hours . After 7 days of rifampin therapy, the medication was discontinued and the pruritus resolved . CONCLUSIONS: This case evaluation demonstrated probable pruritus induced by intravenous rifampin . Clinicians should be alerted that intravenous rifampin, as well as orally administered rifampin, has the potential to cause moderate pruritus.

J Endod, 1995 Mar, 21(3), 109 - 12
Bacterial leakage of mineral trioxide aggregate as a root-end filling material; Torabinejad M et al.; Previous dye leakage studies have shown that mineral trioxide aggregate leaks significantly less than other commonly used root-end filling materials . This study determined the time needed for Staphylococcus epidermidis to penetrate a 3-mm thickness of amalgam, Super-EBA, Intermediate Restorative Material (IRM), or mineral trioxide aggregate (MTA) as root-end filling materials . Fifty-six single-rooted extracted human teeth were cleaned and shaped using a step-back technique . Following root-end resection, 48 root-end cavities were filled with amalgam, Super-EBA, IRM, or MTA . Four root-end cavities were filled with thermoplasticized gutta-percha without a root canal sealer (+ control), and another four were filled with sticky wax covered with two layers of nail polish (- control) . After attaching the teeth to plastic caps of 12-ml plastic vials and placing the root ends into phenol red broth, the set-ups were sterilized overnight with ethylene dioxide gas . A tenth of a microliter of broth containing S . epidermidis was placed into the root canal of 46 teeth (40 experimental, 3 positive, and 3 negative control groups) . In addition, the root canals of two teeth with test root-end filling materials and one tooth from the positive and negative control groups were filled with sterile saline . The number of days required for the test bacteria to penetrate various root-end filling materials was determined . Most samples whose apical 3 mm were filled with amalgam, Super-EBA, or IRM began leaking at 6 to 57 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Transplantation, 1995 Feb 15, 59(3), 382 - 9
Differential effects of the immunosuppressive agents cyclosporine and leflunomide in vivo . Leflunomide blocks clonal T cell expansion yet allows production of lymphokines and manifestation of T cell-mediated shock; Lang R et al.; The effects of leflunomide and CsA on immune responses in vitro and in vivo were investigated . Like CsA, leflunomide inhibited mitogen- or antigen-driven T cell proliferation in vitro . However, leflunomide impaired neither the capability of T cells to produce IL-2 and IL-4, nor the expression of IL-2R, that is, the acquisition of competence . In contrast to CsA, the IL-2-driven growth of secondary T cells was blocked by leflunomide . Cell cycle analyses revealed that activated T cells did not enter S phase of the cell cycle in the presence of leflunomide . Next, the effects of leflunomide and CsA on the T cell response toward the bacterial superantigen (SAg) staphylococcal enterotoxin B (SEB) were analyzed in vivo . SEB-induced early deletion (apoptosis) of a fraction of SEB-reactive V beta 8+ T cells and IL-2R expression were not impaired by either CsA nor leflunomide . On the other hand, both CsA and leflunomide prevented V beta 8-selective clonal T cell expansion and generation of SEB-specific cytolytic activity . In contrast to CsA, leflunomide treatment permitted in vivo SEB-induced production of T cell-derived lymphokines (IL-2 and TNF) . Further, leflunomide failed to protect D-galactosamine-sensitized mice from SEB-induced, T cell-mediated lethal shock, whereas CsA was fully protective . Manifestation of SEB-induced T cell anergy was not impaired by leflunomide . Our results provide evidence that CsA and leflunomide differ significantly in their functional properties to suppress immune responses in that both agents inhibit T cell functions linked to clonal expansion, while leflunomide does not inhibit lymphokine secretion and thus permits lymphokine-mediated immune functions.

J Immunol, 1995 Feb 15, 154(4), 1777 - 85
Superantigen-induced human CD4+ helper/killer T cell phenomenon . Selective induction of Th1 helper/killer T cells and application to tumor immunotherapy; Kuge S et al.; Human CD4+ T cells activated with staphylococcal enterotoxin A (SEA) were fractionated by Percoll discontinuous density gradient centrifugation to enrich SEA-reactive CD4+ T cells . The SEA-reactive CD4+ T cells showed significant cytotoxicity, so-called superantigen-dependent cell-mediated cytotoxicity, against SEA-coated class II-positive tumor cells . During lysis of SEA-coated tumor cells, SEA-reactive CD4+ T cells produced high levels of IL-2 and IFN-gamma but not IL-4 in an Ag-specific manner . The skewing of human CD4+ T cells to Th1-type helper/killer T cells was also demonstrated when SEA-reactive CD4+V beta 5.3+ clonal T cells were cultured with SEA, but not with PHA or OKT3 mAb . Interestingly, the generation of SEA-reactive helper/killer T cells was negatively regulated by IL-4, but up-regulated by IL-12 . The SEA-reactive CD4+ helper/killer T cells were able to generate from PBMC of tumor patients and could be expanded to 10(9) levels in a 7-day culture . The SEA-reactive CD4+ helper/killer T cells were specifically targeted to c-erbB-2 positive human colon cancer cells using SEA-conjugated-anti-c-erbB-2 mAb . These results initially demonstrated that SEA-activated human CD4+ T cells are a Th1 type of Th cell that has both helper and killer functions which may be useful for adoptive tumor immunotherapy in combination with SEA-conjugated antitumor mAb.

Med Klin (Munich), 1995 Feb 15, 90(2), 67 - 71
{Internal medicine complications of ventriculoatrial shunt}; Samtleben W et al.; BACKGROUND: Ventriculo-atrial shunts (VASs) and ventriculo-peritoneal shunts (VPSs) are the symptomatic treatment of choice for hydrocephalus . Bacterial contamination of the atrial part of VASs (usually with Staphylococcus epidermidis) can result in further organ complications, in most instances immune complex mediated glomerulonephritis ("shunt-nephritis") or direct microbial heart valve destruction . PATIENTS AND METHODS: In a retrospective study, we analyzed clinical and laboratory data of 11 patients with VAS associated complications as well as the course of the disease . RESULTS: The following complications were observed: glomerulonephritis (n = 9), glomerulonephritis and aortic valve destruction (n = 1), pulmonary embolism, pulmonary hypertension and tricuspid valve insufficiency (n = 1) . Out of the 11 patients, 8 suffered from unexplained fever . All 11 patients had elevated circulating immune complexes . In 3 of 4 patients initially requiring dialysis, renal function improved which allowed to stop hemodialysis . Renal function also improved in 3 of 5 patients who presented with elevated serum creatinine . Unfortunately, the patient with multiple pulmonary embolisms and tricuspid valve insufficiency died of progressive pulmonary hypertension . CONCLUSION: The prognosis for impaired renal function is good only if the VAS infection is diagnosed early and an immediate surgical and antibiotic treatment leads to an eradication of the underlying chronic infection.

Biochemistry, 1995 Feb 14, 34(6), 2034 - 41
Energetics of denaturation and m values of staphylococcal nuclease mutants; Carra JH et al.; In a continuation of an earlier study {Carra, J., Anderson, E., & Privalov, P . (1994) Biochemistry 33, 10842-10850}, we used differential scanning calorimetry to measure the enthalpy and heat capacity changes of denaturation for 11 mutant forms of staphylococcal nuclease, including the triple mutant {V66L+G88V+G79S} . Several mutant proteins with m- characteristics of guanidinium chloride denaturation were found to denature via a three-state mechanism with increasing temperature . Enthalpy changes for the transitions from the native to intermediate and from the intermediate to denatured states were determined . In the case of the triple mutant, the enthalpy of the second endothermic transition is greater than that of the first . Observation of this second transition provides an explanation for the previously reported large changes in the delta H denaturation for the triple mutant versus wild-type nuclease . The sequence specificity of structure in the intermediate state is discussed with relevance to m values of guanidinium chloride denaturation . The enthalpic level of the intermediate state depends upon the amino acid sequence, suggesting that stabilizing mutations can increase the extent or cohesion of structure present in the intermediate.

Transfus Sci, 1995 Mar, 16(1), 85 - 94
Evidence for proteolytic cleavage of covalently bound protein A from a silica based extracorporeal immunoadsorbent and lack of relationship to treatment effects; Balint JP Jr et al.; Studies were conducted to evaluate the potential cause for release of covalently bound Staphylococcal protein A (SpA) from a silica based extracorporeal immunoadsorbent matrix . In vitro tests revealed that SpA could be detected in human plasma, human serum, and chicken serum upon exposure to the immunoadsorbent matrix which had been treated to remove non-covalently bound SpA . In contrast, only minute quantities of SpA were detected after exposure of a physiologic mixture of purified albumin and immunoglobulin G (IgG) to the immunoadsorbent matrix . Additional tests, employing a cocktail of protease inhibitors and formalin as a general stabilizer and protease inhibitor, revealed significant inhibition of endogenous proteolytic activity present in plasma and serum . Prevention of this proteolytic activity also significantly inhibited the release of covalently bound SpA from the immunoadsorbent matrix upon contact with plasma or serum samples . Further analyses of serum samples from patients with immune thrombocytopenia, chemotherapy associated thrombotic thrombocytopenic purpura-hemolytic uremic syndrome, and breast cancer revealed a lack of association between the quantity of SpA proteolytically released and observed clinical responses or adverse effects experienced during immunoadsorption treatments . These studies indicate that SpA detected in plasma or serum after exposure to the immunoadsorbent is due to inherent endogenous proteolytic activity which cleaves protein fragments from the matrix and that these cleaved SpA fragments do not appear to contribute to the observed clinical responses or adverse effects in treated patients.

Nature, 1995 Feb 2, 373(6513), 438 - 41
Autocrine T-cell suicide mediated by APO-1/(Fas/CD95)
Dhein J, Walczak H, Baumler C, Debatin KM, Krammer PH.
The APO-1/(Fas/CD95) cell surface receptor is a member of the nerve growth factor (NGF)/tumour necrosis factor (TNF) receptor superfamily and mediates apoptosis . Peripheral activated T cells (ATC) from lymphoproliferation (lpr/lpr) mutant mice that express a reduced number of APO-1 receptors have a defect in T-cell receptor (TCR)-induced apoptosis . This suggests that TCR-induced apoptosis involves APO-1 . We tested this hypothesis in various human T cells: (1) malignant Jurkat cells, (2) an alloreactive T-cell clone (S13), and (3) peripheral ATC . TCR triggering through immobilized anti-CD3 antibodies or Staphylococcus enterotoxin B (SEB) superantigen induced expression of the APO-1 ligand and apoptosis in these cells . Anti-CD3-induced apoptosis of Jurkat cells was demonstrated even in single-cell cultures . In all cases apoptosis was substantially inhibited by blocking anti-APO-1 antibody fragments and soluble APO-1 receptor decoys . The APO-1 ligand was found in the supernatant of activated Jurkat cells as a soluble cytokine . We propose that TCR-induced apoptosis in ATC can occur through an APO-1 ligand-mediated autocrine suicide . These results provide a mechanism for suppression of the immune response and for peripheral tolerance by T-cell deletion.

Eur J Biochem, 1995 Feb 1, 227(3), 757 - 71
Sequence analysis by NMR spectroscopy of the peptide lantibiotic epilancin K7 from Staphylococcus epidermidis K7; van de Kamp M et al.; The amino acid sequence of the novel lantibiotic epilancin K7 from Staphylococcus epidermidis K7 was determined by NMR spectroscopy . NMR spectroscopy was used because sequencing by conventional Edman degradation techniques was prohibited by internal sequence blocks owing to the presence of modified residues . Epilancin K7 consists of 31 residues, including two alpha,beta-didehydroalanine (one-letter code U) and two alpha,beta-didehydrobutyrine (O) residues, one lanthionine (A-S-A), two beta-methyllanthionines (A*-S-A), and six lysines . Epilancin K7 has a molecular mass of 3032 +/- 1.5 Da . The amino acid sequence of epilancin K7 was derived from both through-space dipolar proton-proton interactions and through-bond scalar proton-carbon interactions as detected by two-dimensional 1H-NOESY, 1H-ROESY and three-dimensional 1H-TOCSY-NOESY, and by two-dimensional 1H,13C-heteronuclear multiple-bond correlation spectroscopy, respectively . The sequence is as follows: {sequence: see text} The N-terminal residue X partly resembles an alanine but its exact nature is unclear . The organization of the sulfide-bridge-containing (beta-methyl-)lanthionines was revealed by 1H-NMR and 1H,13C-NMR spectroscopy . Epilancin K7 has a linear structure and a high positive net charge, and therefore is classified as a type-A lantibiotic . NMR analysis of a degraded though still active form of epilancin K7 showed that two N-terminal residues of epilancin K7 were missing, owing to decomposition at the alpha,beta-didehydro alanine at position 3; it was called the epilancin K7-(3-31)-peptide (peptide fragment of epilancin K7 consisting of positions 3-31) . The usefulness of three-dimensional 1H-TOCSY-NOESY, and two-dimensional 1H,13C-heteronuclear multiple-bond correlation spectroscopy at natural abundance for the study of (modified) polypeptides is demonstrated.

Ann Thorac Surg, 1995 Feb, 59(2), 348 - 51
The role of thoracoscopy in the diagnosis of interstitial lung disease; Krasna MJ et al.; A study was undertaken to evaluate the safety and efficacy of thoracoscopic lung biopsy for interstitial lung disease . The relation between operative findings, pathologic findings, and preoperative computed tomographic scan findings was examined . Twenty-six patients, 10 male and 16 female, underwent thoracoscopic lung resection to diagnose interstitial lung disease . Sixteen patients were outpatients for an elective procedure; 10 were inpatients including 2 who were ventilator dependent . The mean length of operation was 54 minutes and the mean length of chest tube duration, 1.3 days . There were no deaths . Staphylococcal pneumonia developed in 1 patient postoperatively . One patient with systemic pulmonary hypertension was ventilator dependent for 48 hours . A double-lumen endotracheal tube was used in all but 2 patients . Twelve-millimeter trocar ports were used to allow easy interchange of staplers and endoscopic instruments . Biopsy of at least two lobes was performed in each patient with resection of a piece of grossly abnormal lung . A single chest tube was left routinely . The pathologic diagnosis was usual interstitial pneumonitis in 7 patients . Four patients had interstitial fibrosis and 4, granulomas . Three patients had diffuse alveolar damage and 3, Wegener's granulomatosis . Two patients had bronchiolitis obliterans with organizing pneumonia . One patient each had lymphangioleiomyomatosis, eosinophilic granuloma, and cytomegalovirus . Sixteen patients underwent preoperative computed tomographic scanning . The scans were assessed by 2 radiologists who were blinded to the surgical results . Computed tomography accurately predicted the site of disease in most instances . Four patients had at least one lobe with no evidence of disease on computed tomography but with interstitial lung disease found thoracoscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

J Infect Dis, 1995 Feb, 171(2), 486 - 9
Divergent effects of zinc on different bacterial pathogenic agents; Driessen C et al.; Zinc is essential for immunologic function; therefore, it has been postulated that elevated serum levels of zinc might lead to improved immune responses . However, it is not known whether or how serum zinc levels contribute to a clinically relevant mechanism of immunologic activation . In our studies with human peripheral blood mononuclear cells and whole blood, the zinc level selectively enhanced the biologic activity of endotoxin . The combination of nonstimulatory doses of lipopolysaccharide (LPS) and nonstimulatory concentrations of zinc led to the secretion of large amounts of interleukin (IL)-1 beta . In contrast, zinc levels specifically down-regulated monocyte activation caused by some superantigens, staphylococcal enterotoxin A and E and Mycoplasma arthritidis--derived superantigen, but not toxic shock syndrome toxin-1 . This demonstrates that zinc levels control IL-1 beta secretion after both LPS and superantigen challenge within a clinically relevant range of concentrations . Our data suggest that the indications and contraindications for clinical zinc supplementation should be reconsidered.

Pediatrics, 1995 Feb, 95(2), 225 - 30
Coagulase-negative staphylococcal bacteremia among very low birth weight infants: relation to admission illness severity, resource use, and outcome; Gray JE et al.; OBJECTIVE . To examine the impact of admission-day illness severity on nosocomial bacteremia risk after consideration of traditional risk determinants such as birth weight and length of stay . METHODS . The hospital courses for 302 consecutive very low birth weight (less than 1500 g) infants admitted to two neonatal intensive care units were examined for the occurrence of nosocomial coagulase-negative staphylococcal bacteremia . Using both cumulative incidence and incidence density as measures of bacteremia risk, we explored the relation between illness severity (as measured by the Score for Neonatal Acute Physiology {SNAP}) and bacteremia both before and after birth weight adjustment . In addition, the effect of bacteremia on hospital resource use was estimated . RESULTS . Coagulase-negative staphylococcus was the most common pathogen noted in blood cultures drawn at 48 hours after admission or later . It was isolated on at least one occasion in 53 patients (cumulative incidence of 17.5 first episodes per 100 patients) . These episodes occurred during 7652 days at risk, giving an incidence density of 6.9 initial bacteremias per 1000 patient-days at risk . As expected, when compared with the nonbacteremic group, bacteremic patients were of lower birth weight (888 +/- 231 vs 1127 +/- 258 g; P < .01) and gestational age (26.4 +/- 2.1 vs 28.9 +/- 2.8 weeks; P < .01) . In addition, these patients were more severely ill on admission (SNAP 17.3 +/- 6.5 vs 12.2 +/- 5.8; P < .01) . Even after birth weight stratification, the risk of bacteremia by both measures increased with higher SNAP scores . For example, among infants with birth weights greater than 1 kg, 25% of the most severely ill patients (SNAP 20 and higher) experienced at least one bacteremic episode, whereas the rates seen in infants with intermediate (SNAP 10 to 19) and low illness severity (SNAP 0 to 9) were 8.6% and 3.0%, respectively (chi 2 for trend = 7.25; P < .01) . Multivariate linear regression showed that bacteremia was associated with a prolongation of neonatal intensive care unit stay of 14.0 +/- 4.0 days (P < .01) and an increase in hospital charges of $25,090 +/- 12,051 (P < .05), even after adjustment for birth weight and admission-day SNAP . CONCLUSIONS . Nosocomial coagulase-negative bacteremia is an important complication among very low birth weight infants . Assessment of illness severity with SNAP provides information regarding nosocomial infection risk beyond that available from birth weight alone.

J Immunol, 1995 Feb 1, 154(3), 1024 - 31
Inhibition of antigen-specific T cell activation by staphylococcal enterotoxins; Dowd JE et al.; The staphylococcal enterotoxins SEA, SEB, SEC2, and TSST-1 bind to MHC class II molecules and stimulate polyclonal T cell populations on the basis of the expression of responsive TCR V beta domains . CL-1 is a human T cell clone that is specific for a peptide derived from influenza hemagglutinin (HA 307-319) presented in the context of HLA-DR1 . CL-1 expresses the TCR V beta 13.1 domain, and does not respond to SEA, SEB, or TSST-1 . This T cell was used to test the effect of nonstimulatory staphylococcal enterotoxins on a response to antigenic peptide . These toxins inhibit peptide-specific activation of CL-1 in a concentration-dependent manner . These toxins also inhibit the response of an HLA-DR1-specific alloreactive T cell clone . This inhibition seems to be a result of impaired access of TCR to the MHC/peptide complex rather than negative signaling by toxin via class II interaction or induction of T cell anergy . SEA, but neither SEB nor TSST-1 impedes avidin access to a biotin group attached to the amino terminus of HA 307-319 . SEA partially impairs access of avidin to HA peptide biotinylated at residue 313, but is unable to inhibit avidin access to biotin at residue 318 . This demonstrates that SEA binds to HLA-DR molecules that have also bound the antigenic peptide and suggests a topology for the interaction of SEA with class II, whereby the toxin interferes with peptide/MHC-TCR contact.

Infect Immun, 1995 Feb, 63(2), 423 - 9
Staphylococcal enterotoxin A and toxic shock syndrome toxin compete with CD4 for human major histocompatibility complex class II binding; Bavari S et al.; We have examined the role of the CD4 molecule in primary T-lymphocyte responses to the staphylococcal enterotoxins SEA, SEB, SEC1, and the toxic shock syndrome toxin TSST-1 . Proliferating cells were predominantly CD4+; however, the responses to SEA and TSST-1 were most sensitive to inhibition by the anti-CD4 antibody Leu-3a . T-lymphocyte responses to the bacterial superantigens were inhibited by site-directed mutations of residues in the DR beta membrane-proximal domain (DR beta 2) that are also known to be important for interactions with CD4 . SEA and TSST-1 binding to DR was reduced by the DR beta 2 mutations and by competition with soluble recombinant CD4 . We propose that bacterial superantigens sequentially, or simultaneously with CD4, stabilize complexes of T-cell antigen receptors and major histocompatibility complex class II molecules . The superantigen qualities of these toxins may be due, in part, to a molecular mimicry of CD4 and other adhesion molecules.

Prenat Diagn, 1995 Feb, 15(2), 149 - 54
Fetal erythroblasts from maternal blood identified with 2,3-bisphosphoglycerate (BPG) and in situ hybridization (ISH) using Y-specific probes; Von Koskull H et al.; Different types of fetal nucleated cells can be found in maternal blood, providing the possibility of non-invasive prenatal diagnosis . For this purpose, we have studied fetal erythroblasts . We discovered that haemoglobin-containing cells treated with 2,3-bisphosphoglycerate (BPG) can be visualized by a peroxidase reaction, which at the same time visualizes an in situ hybridization (ISH) signal, specific for the X, Y or 21 chromosome . In order to prove that the BPG-positive cells were erythroid, an anti-glycophorin A (GPA) antiserum combined with a staphylococcal rosette technique was used . To enrich for erythroblasts, leukocytes were depleted from maternal blood by treatment with anti-CD45 monoclonal antibody and passage over an anti-mouse IgG-coated glass bead column . To evaluate the potential of the method for clinical use, we studied maternal blood samples from 18 women referred to us for prenatal diagnosis between 6 and 19 weeks of gestation . Erythroblasts were found in 13 out of 14 normal pregnancies . Erythroblasts with a Y-signal were found as early as 9 weeks of gestation, but at 6 weeks the Y-signal was seen in BPG-negative cells only . These cells showed an epithelioid morphology indicating that they were cytotrophoblasts . The BPG-ISH method provides a simple technique for identifying erythroblasts and simultaneously visualizing a desired probe.

Eur J Nucl Med, 1995 Feb, 22(2), 139 - 47
Immunoscintigraphy with antigranulocyte monoclonal antibodies for the diagnosis of septic loosening of hip prostheses; Boubaker A et al.; To determine the value of immunoscintigraphy (IS) with antigranulocyte monoclonal antibodies (Mab) in the diagnosis of subacute or chronic infection of hip prostheses, we prospectively studied 57 patients (23 women and 34 men; age 29-92 years, mean 72.7 years) sent to our institution in the past 6 years for clinical suspicion of septic loosening of a hip prosthesis . Nineteen patients had bilateral prostheses and one of them was studied twice . A total of 78 prostheses were examined . All patients had three-phase bone scans followed by IS with technetium-99m antigranulocyte Mab BW 250/183 . Intervals between bone scans and IS varied from 2 days to 4 weeks . Final diagnosis was assessed by culture in 48 cases (articular puncture or intraoperative sampling) and by clinical follow-up of at least 8 months in 30 cases . Twelve prostheses were considered septic and 66 non-septic . The overall sensitivity and specificity were 92% and 64% respectively for bone scans, 67% and 75% for IS and 67% and 84% for both modalities together . In three cases, IS was doubtful and the final clinical diagnosis was negative for infection . False-positive results were observed in the presence of massive loosening of the prosthesis or in association with metaplastic peri-articular bone formation . In three of the four false-negative results, infection was proven only after enrichment of the culture, and the bacterium was Staphylococcus epidermidis . In 12/33 (36%) positive bone scans IS allowed the diagnosis of infection to be excluded . Overall accuracy of both modalities together was 81% and the negative predictive value was 93%, which compares favourably with the results reported for other non-invasive methods.

Acta Paediatr, 1995 Feb, 84(2), 226 - 7
Pyomyositis caused by vigorous exercise in a boy; Jayoussi R et al.; We report on a 12-year-old boy who suffered from staphylococcal pyomyositis in both arms following vigorous exercise . No trauma or skin lesions were observed . In non-tropical areas, the etiology of pyomyositis can be related to muscle injury caused merely by exercise, without any other predisposing factors.

Immunology, 1995 Feb, 84(2), 272 - 7
Zinc regulates cytokine induction by superantigens and lipopolysaccharide; Driessen C et al.; Zinc is known to be greatly involved in the regulation of immune functions . Pharmacological zinc supplementation, leading to serum zinc concentrations of more than 0.025 mM, has often been suggested to improve immune responses . However, the exact influence of elevated zinc level on immune functions has not yet been investigated . We found that zinc level selectively enhances cytokine induction by lipopolysaccharide (LPS) in a concentration-dependent fashion: as little as 0.0125 mM supplemental zinc led to nearly 50% elevated interleukin-1 beta (IL-1 beta) levels both in polymorphonuclear cells (PBMC) and whole-blood cultures . The secretion of interferon-gamma (IFN-gamma) could be increased more than 10-fold by 0.1 mM zinc . This could not be observed during stimulation with phytohaemagglutin (PHA) . In contrast, zinc levels concentration-dependently down-regulated monocyte activation caused by the superantigens, staphylococcal enterotoxins A and E (SEA, SEE, more than 90% down-regulation by 0.1 mM zinc), the Mycoplasma arthritidis-derived superantigen (MAS), but not toxic shock syndrome toxin-1 (TSST-1), while T-cell response remained unaffected . This was not the result of chemical degradation of the superantigens . We assume that zinc concentration regulates interactions between SEA, SEE and MAS, but not TSST-1 and their major histocompatibility complex (MHC) class II-binding sites . Our data demonstrate that zinc levels control the secretion of IFN-gamma and monokines after both LPS and superantigen challenge within a clinically relevant range of concentrations . This reveals new perspectives and indications for zinc supplementation and also indicates potential risks of therapeutic application of zinc.

Vet Microbiol, 1995 Feb, 43(2-3), 151 - 9
Chloramphenicol resistance in Staphylococcus intermedius from a single veterinary centre: evidence for plasmid and chromosomal location of the resistance genes; Schwarz S et al.; A total of seven Staphylococcus intermedius cultures isolated from cases of canine pyoderma were investigated for the genetic basis of chloramphenicol resistance (Cmr) . All of these S . intermedius isolates mediated Cmr via the expression of the Cm-inactivating enzyme chloramphenicol acetyltransferase (CAT); the respective cat genes were found to be located on small multicopy plasmids of 3.1 to 4.1 kb in four of the seven cultures . The four Cmr plasmids, designated pSCS20-23, differed upon restriction endonuclease mapping . Hybridization experiments identified all of them to belong to the pC221-family of staphylococcal Cmr plasmids . The expression of all four plasmid-encoded cat genes was inducible with chloramphenicol . The remaining three S . intermedius isolates also harboured an inducible cat gene of the pC221-type which, however, was found to be located in the chromosomal DNA . These differences in the subcellular localisation and consequently in the number of cat gene copies per S . intermedius cell had no influence on the MIC values of Cm exhibited by the respective S . intermedius isolates.

J Biomed Mater Res, 1995 Feb, 29(2), 247 - 56
Blood-biomaterial interactions in a flow system in the presence of bacteria: effect of protein adsorption; Sapatnekar S et al.; An in vitro continuous flow system with whole human blood was used to study blood-biomaterial interactions on a base polyurethane and three modified surfaces in the presence and absence of circulating Staphylococcus epidermidis . We hypothesized that the composition of the protein layer adsorbed on the surface of the biomaterial would influence the response of blood components and bacteria . We examined the test surfaces for adsorption of nine plasma proteins and adsorption profiles differed on the four surfaces . The positively charged surface, UC, adsorbed significantly higher amounts of fibronectin (P < .01), von Willebrand factor (P < .01), and fibrinogen (P < .05) than the other materials . As a consequence of increased adsorption of these adhesive proteins, the adhesion of platelets and bacteria was greater on UC than on any other surface . On the base polyurethane, BC, and the negatively charged surface, UA, protein adsorption was low, and these materials were largely free of adherent blood cells and bacteria . The heparinized surface, UH, adsorbed higher quantities (P < .01) of Hageman factor and high molecular weight kininogen relative to the other surfaces . Platelet adhesion, and surface coagulation were prominent on UC, and may have contributed to increased bacterial adhesion on this surface . In the presence of circulating bacteria, adsorption was generally lower than in the absence of bacteria . The pattern of protein adsorption was largely unaffected by the strain of circulating bacteria, but platelet responses (adhesion and activation) were greater in the presence of slime-producing S . epidermidis as compared to the non-slime-producing strain, suggesting that slime may have a direct activating effect on platelets.

Int Immunol, 1995 Feb, 7(2), 295 - 304
Co-stimulation lowers the threshold for activation of naive T cells by bacterial superantigens; Muraille EM et al.; Staphylococcus enterotoxins bind class II MHC molecules on antigen presenting cells (APC) and stimulate T cells expressing appropriate V beta gene products . Although the role of non-TCR associated co-stimulatory receptors during antigen-specific T cell stimulation has been clearly established, the involvement of co-stimulatory activity in T cell activation by superantigens has been the matter of controversy . In this report, we examine the role of co-stimulation provided by selected APC populations in the response to bacterial exotoxins (staphylococcal enterotoxin A, staphylococcal enterotoxin B and toxic shock syndrome type 1) . We demonstrate that the APC population able to activate naive T cells to IL-2 production is heterogeneous, comprising both adherent (presumably dendritic) and non-adherent (mostly B lymphocytes) cells . By stimulating naive T cells in the presence of graded doses of superantigens, we have observed that half-maximal IL-2 production was achieved at lower doses of superantigens in the presence of dendritic cells . Similarly, addition of antibodies to CD28 or B7.1-transfected cell lines increased the sensitivity of naive T cells to lower doses of superantigens . These observations indicate therefore that superantigens can be presented to naive T cells by APC displaying distinct levels of co-stimulatory activity, although with different efficacy . Thus, naive T cells are sensitive to CD28-mediated co-stimulation during superantigen-mediated responses but IL-2 production can be induced by high doses of superantigens in the presence of APC expressing weak co-stimulatory activity . These observations are compatible with the hypothesis that CD28-mediated signals participate in T cell activation by lowering T cell sensitivity to TCR ligands.

Baillieres Clin Rheumatol, 1995 Feb, 9(1), 193 - 200
Bacterial infections: pyomyositis; McGill PE; Staphylococcal pyomyositis is an important and common condition in many areas in the tropics . The cause is probably multifactorial and includes damage to the skeletal muscle in the presence of staphylococcal bacteraemia, with or without depressed immunity . In Africa, there are indications of an increased prevalence in association with HIV infection in young adults in whom multiple and recurrent abscesses are common . Long bone osteomyelitis is an important differential diagnosis in these patients . Timely surgical drainage and antibiotics leads to resolution . Delayed diagnosis is associated with disseminated disease and septic cardiorespiratory complications.

J Chemother, 1995 Feb, 7(1), 26 - 9
Staphylococcus epidermidis isolation and antibiotic resistance in a neonatal intensive care unit; Fanos V et al.; Bacterial ecology was studied in 1114 newborns (355 at term, 759 preterm) admitted to a neonatal intensive care unit (NICU) during a three year period . Bacterial samples were taken in each newborn from external ear canal, pharynx and eyes in all patients, and from endotracheal tube, umbilical catheter and blood in selected patients . The predominant flora was characterized by gram-positive microorganisms (63.53%), Staphylococcus epidermidis representing 34.68% of all isolated strains . S . epidermidis isolation increased significantly with time (p < 0.002) and was highest in summer . The percentage of S . epidermidis resistant strains to oxacillin (63.8%) and to amikacin (17.8%) was high . This is the antimicrobial combination we commonly employ as empirical treatment of suspected bacterial infection in our NICU . Knowledge of characteristics of local microbial flora seems important in order to optimize preventive and therapeutic policies for neonatal infections.

Antibiot Khimioter, 1995 Feb, 40(2), 46 - 50
{Microbiologic diagnosis of mixed anaerobic and aerobic surgical infections}; Zubkov MN et al.; One hundred and twelve patients with purulent affections of the abdominal and thoracic cavities as well as soft tissues and 49 patients with suspected generalization of the infection were examined . It was shown that the position of mixed anaerobic and aerobic infections in various groups of the patients could reach 70-80 per cent while the species composition of the pathogens was close to that of the normal microflora in the affected organ or its region . Bacteroides spp . played the main role in the development of anaerobic sepsis and conditioned a distinctive process . There was observed synergism of Escherichia coli and Staphylococcus spp . and they were shown to serve as indicators of the anaerobic infection due to these species in definite biotops . The use of GLC markedly lowered the time required for diagnosis of anaerobic infections which provided rational chemotherapy at the early stages of the disease with an account of the data on monitoring of the suppuration causative agent.

Toxicon, 1995 Feb, 33(2), 241 - 5
Isolation of peptides homologous to domains of human alpha 1B-glycoprotein from a mongoose antihemorrhagic factor; Qi ZQ et al.; Thirteen peptides, homologous to one of the five domains of human alpha 1B-glycoprotein (alpha 1BG), were isolated from a mongoose antihemorrhagic factor (AHF1); four of them were generated by BrCN cleavage, and three and six peptides by the digestions with lysyl endopeptidase and staphylococcal protease V8, respectively . The purified peptides covered 75.9% of the whole sequence of human alpha 1BG (359/474 residues) and showed 46.4% identity (167/360 amino acids) with the sequence of human alpha 1BG including cysteine residues forming disulfide linkages . One of the sugar binding sites of human alpha 1BG was also conserved in AHF1 . These results suggest that AHF1 is a protein homologous to human alpha 1BG and a supergene family of immunoglobulins.

Toxicon, 1995 Feb, 33(2), 229 - 39
The primary structure of a hemorrhagic factor, HR2b, from the venom of Okinawa habu (Trimeresurus flavoviridis); Iha M et al.; The complete amino acid sequence of a hemorrhagic factor, HR2b, from the venom of Okinawa habu was determined . The hemorrhagic factor was fragmented by CNBr cleavage, trypsin, staphylococcal protease V8 and lysyl endopeptidase digestions . The resulting peptides were purified on high performance chromatography, and sequenced by Edman degradation . HR2b was composed of 204 amino acids with pyroglutamyl residue at the amino terminus, and the calculated mol . wt based on the amino acid composition was 23,335 . There are three disulfide linkages in the primary structure . The consensus sequence (His-Glu-Xaa-Xaa-His) for zinc-binding site of zinc-requiring metalloproteinases was found in the structure . The primary structure of HR2b shows a significant similarity with that of HR2a of Amami habu venom; 98.5% identity.

Jpn Circ J, 1995 Feb, 59(2), 103 - 11
Role of protein kinase C in relationship between Ca2+ and contractile elements in rat alpha-toxin-permeabilized mesenteric artery; Sasajima H et al.; Phorbol ester, which activates protein kinase C (PKC), modulates vasoconstrictor-induced tension in vascular smooth muscle . Recently, Staphylococcal aureus alpha-toxin, which produces too small pores in the plasma membrane to allow passage of proteins, such as PKC, is used to investigate the signal transduction system in vascular smooth muscle cells . In order to elucidate the role of PKC on vascular smooth muscle contraction, we examined whether PKC activation influences the relationship between intracellular Ca2+ ({Ca2+}i) and tension in Wistar rat superior mesenteric artery (SMA) using vascular smooth muscle permeabilized with Staphylococcal alpha-toxin . {Ca2+}i was clamped at specified values (10(-8.5)-10(-4) mol/L) using EGTA-Ca2+ buffer . In alpha-toxin non-treated rings of SMA, isometric tension was evoked by 10 mmol/L caffeine and 10-30 mmol/L external potassium (high K+) in the absence or presence of phorbol 12, 13-dibutyrate (PDBu), a PKC activator, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), and staurosporine (PKC inhibitors) . PDBu significantly augmented caffeine- and high K(+)-evoked contractions . H-7 and staurosporine significantly attenuated caffeine- and high K(+)-evoked contractions augmented by PDBu . Moreover, H-7 significantly suppressed high K(+)-induced contraction in the absence of PDBu . In alpha-toxin permeabilized artery, PDBu shifted the {Ca2+}i-force relationship curve to the left . These results suggest that PKC activates vascular smooth muscle contraction by increasing the sensitivity of the contractile apparatus to Ca2+.

Cancer Res, 1995 Feb 1, 55(3), 623 - 8
Antibody-targeted superantigens induce lysis of major histocompatibility complex class II-negative T-cell leukemia lines; Ihle J et al.; CTLs bearing certain T-cell receptor V beta-regions are directed by the bacterial superantigen Staphylococcus enterotoxin A (SEA) to lyse MHC class II-positive cells . In order to extend superantigen-dependent cytotoxicity to MHC class II-negative carcinoma cells, covalent conjugates of superantigen and mAbs against surface markers of these cells have been used . We now describe a novel strategy which allows rapid selection of mAb suitable for superantigen targeting against MHC class II-negative tumor cells . A recombinant fusion protein of protein A and SEA binding to the mAbs CD7 or CD38 was able to mediate T cell-dependent lysis of MHC class II-negative Molt-4 and CCRF-CEM acute lymphatic leukemia cell lines . Lysis was dose dependent and correlated with E:T cell ratio . In contrast, SEA alone did not induce any significant lysis . In order to decrease the MHC class II affinity of the protein A-SEA complex, a point mutation was introduced into SEA (protein A-SEA mu9) . The mutated fusion protein had similar potency as protein A-SEA against Molt-4 cells but was 100-fold less active against MHC class II-positive cells . Considering the efficiency and specificity of the mutated SEA protein interacting with mAb in targeting T lymphocytes against MHC class II-negative leukemia cells while only marginally affecting normal MHC class II-positive cells, we suggest the development of SEA-mAb fusion proteins as a potential adjuvant therapy of leukemias.

J Immunol, 1995 Feb 1, 154(3), 1014 - 23
Staphylococcal enterotoxin B activates purified NK cells to secrete IFN-gamma but requires T lymphocytes to augment NK cytotoxicity; D'Orazio JA et al.; Staphylococcal enterotoxin B (SEB) is known for its effects on T lymphocytes; the mechanism of this activation is well studied . Although it is anticipated that there would be effects on other cells in the milieu, the mechanisms of activation of non-T immune cells are not well understood . This report examines the effects of SEB on human NK cells . Incubation of PBMC with SEB augmented NK cytotoxicity against the NK-sensitive targets K562 and Molt-4 and induced activity against the NK-resistant targets, Mel-30 and Raji . The degree of superantigen-augmented killer (SAK) activity induced by the SEB treatment correlated with the concentration of the superantigen in the mixture . Specific neutralizing Abs for lymphokines or monokines added to cultures showed that IL-2, IFN-gamma, or IL-12 each participated in the phenomenon; addition of all Abs together blocked most, but not all, SAK activity . Highly purified T lymphocytes (but not monocytes) supported the development of SAK activity in enriched cultures of CD56+ cells . The SAK activity was blocked by specific Abs to IL-2 and IFN-gamma in cocultures of T cells and CD56+ cells . In addition, SEB induced IFN-gamma secretion from sorted CD56+ cells . These data show for the first time that superantigen directly initiated cytokine synthesis in NK cells, and that SAK activity was induced as a consequence of products secreted from activated T lymphocytes . Such a superantigen effect on NK cells would place the NK lymphocyte in a pivotal role in determining the outcome of infection or immunopathology caused by superantigen exposure.

Infect Immun, 1995 Feb, 63(2), 539 - 46
Regulation of staphylococcal enterotoxin B-elicited nitric oxide production by endothelial cells; LeClaire RD et al.; The effect of staphylococcal enterotoxin B (SEB)-elicited inducible nitric oxide synthase (iNOS) in mouse endothelial cells was investigated . Results showed that SEB stimulated the same level of NO production in gamma interferon (IFN-gamma)-primed cells as did trichloroacetic acid-extracted lipopolysaccharide . The kinetics of induced NO production and expression of mRNA for iNOS differed markedly in endothelial and macrophage cells . Induced endothelial nitrite production was transient and was 15 to 20% of that generated by macrophage cells; mRNA levels peaked by 2 h and then steadily declined, whereas macrophage message levels continually increased . The ability of endothelial cells to produce SEB-induced NO depended on priming with IFN-gamma, although detectable mRNA could be elicited by SEB alone . Induction of endothelial iNOS mRNA was inhibited by cycloheximide, which indicated a requirement for de novo protein synthesis . Niacinamide and interleukin-10 significantly reduced SEB-induced endothelial NO production . Both are reported to affect IFN-gamma-induced class II major histocompatibility complex (MHC) expression on antigen-presenting cells . Niacinamide reduced iNOS mRNA levels and markedly reduced IFN-gamma induction of endothelial class II MHC surface antigen . Interleukin-10 did not consistently reduce iNOS mRNA expression and had no effect on IFN-gamma induction of endothelial class II MHC surface antigen . These results suggest that SEB interacts with IFN-gamma-primed endothelial cells to elicit induced NO and that this induction can be effectively modulated at the receptor or transcriptional level.

Arch Mal Coeur Vaiss, 1995 Feb, 88(2), 241 - 6
{Septicemia and endocarditis related to transvenous pacing leads of pacemakers: surgical indications and results}; Leprince P et al.; Endocarditis of transvenous pacing leads is a rare condition . The authors review a series of 15 patients who developed bacteriologically proven septicaemia and/or endocarditis related to transvenous pacing leads, operated between 1988 and 1993 . The interval between the last manipulation of the pacemaker and the onset of endocarditis was about 6 months . Six patients had had haematoma and/or infection of the pacemaker site . Endocarditis presented with chronic pyrexia (14 cases) associated with septicaemia (6 cases) and chronic local suppuration (1 case) . The interval between the beginning of the pyrexia and the diagnosis was 3.4 months . Echocardiography showed a mass attached to the pacing lead in 8 cases and tricuspid valve vegetations in 4 cases . Blood cultures were positive in 13 patients and local wound swabs identified the organism in 1 patient . The commonest causal agent was the staphylococcus (epidermis in 7 cases, aureus in 4 cases) . Appropriate antibiotic therapy was only effective in 1 case . The surgical indication in 13 cases was persistence of infection associated with pulmonary embolism (3) or tricuspid regurgitation (2) . Complete ablation of the prosthetic material was performed by a peripheral vascular approach (2 cases), by a right atrial approach (1 case) and under cardiopulmonary bypass in 12 cases . The peroperative findings were of tricuspid valve vegetations (4 cases), thrombi on the pacing lead (7 cases) or in the right heart chambers (2 cases) or pulmonary artery (2 cases) . The associated procedures performed under cardiopulmonary bypass were tricuspid valve repair (2 cases) and pulmonary thrombectomy (2 cases) . Temporary and permanent epicardial leads were implanted in 10 patients.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biotechnol, 1995 Jan 31, 38(3), 263 - 8
Production of the chimeric-binding protein, maltose-binding protein-protein A, by gene fusion; Kobatake E et al.; A fusion protein between maltose-binding protein (MBP) and staphylococcal protein A (SpA) was genetically produced . The gene fusion plasmid, pMALPA2, was constructed by inserting the protein A gene into an expression vector of maltose-binding protein in frame, and was expressed efficiently in Escherichia coli . The resulting fusion protein of molecular mass 65 kDa, retained the activity of both MBP and SpA (binding capability to amylose and immunoglobulin G) . This chimeric-binding protein was used as an adhesive molecule for immobilization of antibodies to a solid-phase surface for enzyme immunoassay . An enzyme immunoassay was performed with the fusion protein, and human IgG was determined in the concentration range from 10(-4) to 10(-6) g ml-1.

Proc Natl Acad Sci U S A, 1995 Jan 17, 92(2), 364 - 8
Targeting of a nuclease to murine leukemia virus capsids inhibits viral multiplication; Natsoulis G et al.; Capsid-targeted viral inactivation is an antiviral strategy in which toxic fusion proteins are targeted to virions, where they inhibit viral multiplication by destroying viral components . These fusion proteins consist of a virion structural protein moiety and an enzymatic moiety such as a nuclease . Such fusion proteins can severely inhibit transposition of yeast retrotransposon Ty1, an element whose transposition mechanistically resembles retroviral multiplication . We demonstrate that expression of a murine retrovirus capsid-staphylococcal nuclease fusion protein inhibits multiplication of the corresponding murine leukemia virus by 30- to 100-fold . Staphylococcal nuclease is apparently inactive intracellularly and hence nontoxic to the host cell, but it is active extracellularly because of its requirement for high concentrations of Ca2+ ions . Virions assembled in and shed from cells expressing the fusion protein contain very small amounts of intact viral RNA, as would be predicted for nuclease-mediated inhibition of viral multiplication.

J Immunol, 1995 Jan 15, 154(2), 861 - 70
Chlorpromazine amplifies macrophage-dependent IL-10 production in vivo; Tarazona R et al.; Chlorpromazine (CPZ) has potent immunomodulatory effects in vivo; it induces humoral autoimmunity in up to 50% of patients, inhibits delayed-type hypersensitivity reactions, and suppresses lethal immune hyperactivation in animal models of septic shock . Here, we show that in an in vivo model of acute superantigen-driven immune activation, CPZ independently down-regulates the production of various T cell-derived lymphokines (IL-2, IFN-gamma, IL-4, TNF, and GM-CSF) and up-regulates the secretion of IL-10 . Whereas only low, if any, serum IL-10 levels are detectable by ELISA after injection of CPZ, bacterial LPS, or staphylococcal enterotoxin B (SEB) alone, simultaneous administration of CPZ + LPS or CPZ + SEB causes a significant increase in IL-10 production in vivo . CPZ-mediated amplification of the SEB-driven CPZ secretion is accompanied by an enhanced IL-10 mRNA accumulation, as shown by PCR analysis and in situ hybridization . Determination of IL-10 production in mice lacking T cells, B cells, or phagocytes revealed that SEB + CPZ-induced IL-10 was produced by phagocytic cells, but not by lymphocytes, a finding that is in accord with the distribution of splenic cells transcribing the IL-10 gene in response to SEB + CPZ . Moreover, these data indicate that bacterial superantigen can directly stimulate tissue phagocytes, even in the virtual absence of T lymphocytes . The blockade of dopamine type 1 (D1) but not type 2 (D2) receptors abolishes the CPZ effect on IL-10 production . Inhibition of Th1 and Th2 lymphokine production by CPZ is not mediated by dopamine receptors and is independent of IL-10 up-regulation . These findings may explain the mechanism by which CPZ and related drugs enhance humoral autoimmune reactions, block cellular immune responses, and prevent lethal septic shock in vivo.

Blood, 1995 Jan 15, 85(2), 351 - 8
Pulse cyclophosphamide therapy for refractory autoimmune thrombocytopenic purpura; Reiner A et al.; Autoimmune thrombocytopenic purpura (AITP) is generally a chronic disorder in affected adults . Twenty-five percent of these patients will become refractory to routine therapy (corticosteroids and splenectomy), as well as most other available agents . Intravenous pulse cyclophosphamide therapy was used to treat 20 patients with severe refractory AITP who had previously failed to achieve a sustained remission with a mean of 4.8 agents (range 2 to 8) . Patients received 1 to 4 doses (mean 2.0) of 1.0 to 1.5 g/m2 intravenous cyclophosphamide per course . Of the 20 patients treated with pulse cyclophosphamide therapy, 13 patients (65%) achieved a complete response (CR), four (20%) a partial response (PR), and three patients (15%) failed to respond . Of the 13 complete responders, eight have remained in remission with stable platelet counts during followup intervals of 7 months to 7 years (median 2.5 years) . Five patients developed recurrent AITP 4 months to 3 years following a CR . Of these, two patients responded to subsequent courses of pulse cyclophosphamide therapy with current remissions of 1 and 4 years . Of the four patients who obtained a PR, two remain in partial remission after 10 months and 4 years; one relapsed after 18 months and, after retreatment, is still in remission at 6 months . Of the patient characteristics examined, duration of disease was most strongly associated with response to pulse cyclophosphamide . Side-effects of treatment included neutropenia (three patients, one of whom developed staphylococcal sepsis), acute deep venous thrombosis (two patients), and psoas abscess (one patient) . Intravenous pulse cyclophosphamide should be strongly considered in the treatment of patients with refractory AITP . There is a relatively low incidence of side-effects, and it can be administered easily on an out-patient basis.

Arch Vet Pol, 1995, 35(1-2), 63 - 71
Phagocytosis-enhancing effect of lactoferrin on bovine peripheral blood monocytes in vitro and in vivo; Szuster-Ciesielska A et al.; The effect of lactoferrin (LF) on in vitro and in vivo phagocytic ability of bovine blood monocytes was studied . It was demonstrated that bovine LF enhanced in vitro phagocytosis of bacteria and ovine erythrocyte-antibody complexes and increased intracellular killing of Staphylococcus albus . Monocytes of colostrum deprived calves, which were intravenously injected with LF, also exhibited elevated phagocytic properties.

Folia Microbiol (Praha), 1995, 40(3), 271 - 3
The effect of salinomycin and lasalocid on laboratory cultures of Enterococcus faecium and Staphylococcus gallinarum strains; Laukova A et al.; The growth of Enterococcus faecium strains CCM 4231 and EF 26, and Staphylococcus gallinarum SG 31 was inhibited by salinomycin and lasalocid at concentrations of 25 and 50 mg/L . Staphylococcus gallinarum was more sensitive to the additives used than were enterococci . Maximum inhibition (90%) was measured after the growth with the SG 31 strain in the presence of both ionophores . Growth of organisms was more inhibited by salinomycin at 25 mg/L (67.5%) than at 50 mg/L (63%) . The inhibitory effect in enterococcal strains reached after the addition of salinomycin and lasalocid (on average) 63 and 58%, respectively . The CCM 4231 strain was more inhibited by salinomycin as well as by lasalocid than was the EF 26 strain.

Pathobiology, 1995, 63(4), 175 - 87
Increased cAMP and cAMP-dependent protein kinase activity mediate anti-CD2 induced suppression of anti-CD3-driven interleukin-2 production and CD25 expression; Lin J et al.; Anti-CD2 monoclonal antibody (mAb) can act synergistically with anti-CD3 to produce tolerance and diminish the anti-CD3-induced cytokine syndrome . Since interleukin(IL)-2 production and IL-2 receptor (IL-2R; CD25) expression are important determinants of CD3-driven T cell activation, the effects of anti-CD2 on anti-CD3-induced CD25 expression and IL-2 production were analyzed and related mechanistically to CD2-stimulated cAMP signaling with an in vitro model of T cell activation . The anti-CD2 mAb, 12-15, alone had no effect on splenic T cell CD25 expression and IL-2 production, while the anti-CD3 mAb, 145-2C11, caused significant increases in both CD25 expression and IL-2 production . The addition of anti-CD2 inhibited anti-CD3-induced increases in CD25 and IL-2 . The inhibitory signal delivered by anti-CD2 was effective in many forms of T cell activation, since other stimuli which increased CD25, such as concanavalin A, phytohemagglutinin, and Staphylococcal enterotoxin B (SEB), could also be inhibited by anti-CD2 . The inhibitory effect of anti-CD2 on CD25 could not be reversed by high doses of supplemental IL-2 added to the culture . Anti-CD2 increased cytoplasmic cAMP in a dose- and time-dependent manner . Reagents that increased cytoplasmic cAMP such as forskolin, cholera toxin, and 3'-isobutyl-1-methylxanthine could mimic the inhibitory effect of anti-CD2 on anti-CD3-driven CD25 expression . Anti-CD2 also increased the activity of cAMP-dependent protein kinase (PKA) . H8, a PKA antagonist, blocked the inhibitory effect of anti-CD2 on CD25 expression, further confirming the role of PKA in CD2-induced negative signaling . The use of paired agonists to PKA demonstrated that a type I PKA was the preferential enzyme isoform stimulated by CD2 ligation . These findings show that increased cAMP and PKA activity mediate anti-CD2-induced suppression of anti-CD3-driven IL-2 production and CD25 expression, and provide mechanisms for anti-CD2-induced immunosuppression and inhibition of the cytokine syndrome associated with anti-CD3 treatment.

J Fr Ophtalmol, 1995, 18(11), 650 - 5
{Search for bacterial contamination of the aqueous humor during cataract surgery with and without local antibiotic prophylaxis}; Saint-Blancat P et al.; PURPOSE: Bacterial contamination of anterior chamber at the end of cataract surgery, was compared between two techniques: extracapsular extraction and phacoemulsification . The effectiveness of preoperative antibiotic eyedrops using Norfloxacine 0.3% (Chibroxine) was also evaluated . METHOD: The study focused on 101 patients grouped according to surgical technique and presence of preoperative antibiotic eyedrops . Conjunctival sampling was made the day prior the surgery, as well as in the operating room, after skin and conjunctival desinfection with povidone iodine in all the patients included in the study . Aqueous humour was collected at the end of surgery . RESULTS: Eight samples out of 101 were positive which represents 7.9% of the cases . In 75% of the cases, the anterior chamber aspirate showed a different germ or non-recurrent germ in the second conjunctival sample . None of the included patients developed endophthalmitis . The two most frequent pathogens were Propionibacterium acnes in 62.5% of the cases, and Staphylococcus epidermidis in 50% . Another pathogen was found in a culture environment: Micrococcus roseus . In two samples, two different bacteria grew: Propionibacterium acnes and Staphylococcus epidermidis . Whatever the surgical technique, no statistically significant bacterial contamination was found . There was no significant statistical difference between patients who had local antibiotic eyedrops and those who did not . CONCLUSION: This study confirms the poor reliability of local antibiotic eyedrops to prevent surgical contamination . Furthermore performing an anterior chamber aspirate at the end of the surgery for risk patients would help the physician identify the pathogens involved in endophthalmitis in order to begin antibiotic treatment as soon as possible.

Ciba Found Symp, 1995, 195, 142 - 7; discussion 147-53
Cytokines in immune regulation/pathogenesis in HIV infection; Shearer GM et al.; Two hallmarks of immunopathogenesis in the progression of HIV-infected individuals to AIDS are the loss of T helper (Th) cell function in response to antigens and the critical reduction in CD4+ T cell numbers . It is probable that these two phenomena are related . We observed that: (1) the failure to detect antigen-stimulated Th cell responses in vitro correlates with increased pokeweed mitogen/staphylococcal enterotoxin B (P/S)-stimulated and antigen-stimulated T cell death; and (2) both of these events are similarly modulated by immunoregulatory cytokines . Interleukin 2 (IL-2) and IL-12 (Th1-type cytokines), as well as antibodies to IL-4 and IL-10 (which are Th2-type cytokines) restore in vitro Th cell responses to recall antigens such as influenza virus and HIV envelope synthetic peptides (env) . P/S-induced T cell death affects both CD4+ and CD8+ T cell subsets, whereas death induced by stimulation with env affects only CD4+ T cells . In both examples, Th1-type cytokines and antibodies to Th2-type cytokines protect against T cell death . In contrast, IL-4 and IL-10 do not protect against death, and anti-IL-12 antibody can enhance T cell death . Our findings indicate that the loss of Th cell function and increased T cell death seen in vitro are correlated, and that in vivo HIV infection gives rise to inappropriate cytokines resulting in immune dysfunction and immunopathogenesis.

Pneumoftiziologia, 1995 Jan-Jun, 44(1-2), 63 - 6
{Pulmonary manifestations in a case of toxic shock syndrome}; Stoica R; The evolution of a toxic shock syndrome in a young adult has yielded particular pleural and lung lesions concomittantly with phenomena of insufficiency of other organs and systems (kidney, digestive, CNS a.o.) . X-ray aspect of bilateral bronchopneumonia with small diffuse infiltrates are noted, together with an aspect of bilateral spontaneous pneumothorax, which appeared successively with a tendency to relapse . Hemodynamic alterings during the acute phase of the syndrome initiated by staphylococcal exotoxin are similar to those during the initial phase of the septic shock, requiring a careful monitoring during the treatment.

Scand J Infect Dis, 1995, 27(6), 575 - 80
Molecular epidemiology of Staphylococcus epidermidis isolates in a hematological unit during a 4-month survey; Lyytikainen O et al.; In order to study the strain variety, clonal stability and epidemiology of Staphylococcus epidermidis, isolates from all bacterial cultures taken when clinically indicated in 2 wards of the hematological unit of Helsinki University Hospital, during a 4-month period, were characterized by 3 typing methods: antibiogram, plasmid profile and ribotype . A total of 141 distinct S . epidermidis colonies, from 28 blood cultures and 37 cultures from other sources in 32 patients were studied . Plasmid profiles and ribopatterns revealed 47 different strains of which 16 were bacteremic . One of these strains caused bacteremia in 4 different patients over a 3-month period and it was isolated from blood on 7 different sampling occasions . The occurrence of this clone was constant; it was usually found in both of 2 blood culture bottles inoculated (6/7 pairs) and dominated among the 17 distinct S . epidermidis colonies studied from the positive bottles (94% of the total) . The clones causing bacteremias in the 2 wards were distinct . These findings indicate that certain clones of S . epidermidis can predominate in hematological wards and that nosocomial transmission of S . epidermidis strains may occur among patients, particularly within the same ward.

Acta Haematol, 1995, 94(4), 196 - 8
Agranulocytosis in rheumatoid arthritis associated with long-term flucloxacillin for staphylococcal osteomyelitis; Burton IE et al.; Agranulocytosis and septicaemia developing in a patient with rheumatoid arthritis after 3 years' intermittent treatment with diclofenac, cimetidine and flucloxacillin for staphylococcal osteomyelitis is described . Treatment with recombinant granulocyte-colony-stimulating factor and high-dose methylprednisolone had no effect on the neutropenia which resolved on stopping all drug therapy . Relapse of agranulocytosis followed reintroduction of flucloxacillin and cimetidine 3 months later.

J Inflamm, 1995, 45(3), 183 - 92
Superantigen-induced transcriptional activation of the human TNF gene promoter in T cells; Kramer B et al.; Superantigens, such as staphylococcal enterotoxins (SE), cause food poisoning and shock, which is mediated at least in part by TNF . We have examined the mechanism of superantigen-induced activation of the TNF gene promoter in primary human peripheral blood T lymphocytes and in the leukemic T cell line Jurkat . Like stimulation of the T cell receptor complex through CD3 ligation, SEB induces binding of nuclear proteins to Egr- and Jun/ATF-related consensus sequences present between nucleotides -170 and -100 of the TNF promoter 5' flanking region . By cotransfection of Jurkat T cells with constructs containing TNF promoter deletion mutants linked to a CAT reporter gene, it is shown that superantigens can activate transcription from these two adjacent ETs and Jun/ATF binding elements . Superantigen-induced binding of Egr-1 and Jun/ATF is markedly reduced by okadaic acid, suggesting that phosphatases are involved in the signaling of SE . When compared to CD3 ligation, superantigens activate the TNF promoter more potently, which is likely due to costimulatory signals provided by superantigen presenting cells.

Retina, 1995, 15(5), 428 - 32
Vancomycin levels after intravitreal injection . Effects of inflammation and surgery; Aguilar HE et al.; OBJECTIVE: Vancomycin hydrochloride for intraocular injection is the drug of choice for the treatment of suspected gram-positive endophthalmitis . To study its intraocular pharmacokinetics, we injected vancomycin into the vitreous cavity of phakic, aphakic, and aphakic-vitrectomized rabbit eyes and determined its rate of clearance . Inflamed eyes were compared to control eyes in each group . METHODS: Three groups of eyes were prepared . The eyes in Group 1 were phakic, the eyes in Group 2 had the lens removed, and the eyes in Group 3 had both the lens and central vitreous removed . Each group was subdivided into a control group and a group made inflamed by injection of heat-killed Staphylococcus epidermidis . Vancomycin hydrochloride 1 mg in 0.1 cc of diluent was injected into the midvitreous cavity and samples obtained at 2 or 3, 8, 24, and 48 hours after injection . Vancomycin concentrations were measured and clearance rates were calculated for each of the groups . RESULTS: Vancomycin was cleared substantially faster from aphakic-vitrectomized eyes (half-life 9.0 hours) and aphakic eyes (half-life 8.9 hours) than phakic eyes (half-life 25.1 hours) . Inflammation increased the rate of elimination of vancomycin only in the aphakic group . CONCLUSIONS: Clearance of vancomycin from the phakic eye is prolonged compared to that of beta-lactam antibiotics, an important pharmacokinetic advantage in treating endophthalmitis . Its clearance from aphakic-vitrectomized eyes is dramatically faster than from phakic eyes and is similar to that of other antimicrobial agents.

J Ocul Pharmacol Ther, 1995 Fall, 11(3), 349 - 59
The treatment of endophthalmitis after cataract surgery: review of 26 cases; Parkkari M et al.; A retrospective study of 26 cases of postoperative endophthalmitis occurring after extracapsular cataract extraction and lens implantation was conducted between January 1989 and December 1992 . Twenty-one of the 26 cases (81%) were diagnosed within two weeks after surgery . The most common organism isolated was Staphylococcus saprophyticus (23%) . After the bacterial samples had been taken, cefotaxime or gentamicin was injected into anterior chamber and vitreous space . The therapy regimen consisted of topical tobramycin, Polysporin (polymyxinsulfate, neomycinsulfate and gramicidin), dexamethasone and atropine and parabulbar cefotaxime, netilmicin and betamethasone . Cefotaxime or, alternatively, clindamycin, ciprofloxacin and prednisolone were given systemically . The final visual acuity of 20/40 or more was obtained in 15 cases (58%) . Twenty-one patients (81%) achieved a visual acuity of 20/400 or better . Three patients had no light perception after the treatment of endophthalmitis . Silk as suture material in wound closure and rupture of the posterior capsule were found to be risk factors in the development of endophthalmitis . The results of the present study show that effective drug treatment also results in good visual outcome.

Acta Clin Belg, 1995, 50(6), 326 - 34
{Epidemiological study on neonatal septicemia and meningitis in Belgium}; Walckiers D et al.; Epidemiological data were collected for 219 cases of neonatal septicaemia and/or meningitis . In two thirds of the cases the diagnosis was made in newborns aged 4 days or less . Risk factors related to the delivery, to the health status of the infant at birth and to pregnancy were mentioned respectively in 74, 63 and 36% of the cases . Half of the newborns given antibiotics in prophylaxis received an association of aminopenicillin and aminoside . In more than four fifths of the cases the bacterium was only isolated from blood . In newborns aged 4 days or less at the time of diagnosis, S . agalactiae was the most frequently isolated agent (50%) . In newborns aged 5 days or more at the time of diagnosis, E . coli was the most frequent isolate (23%), followed by S . agalactiae and coagulase-negative Staphylococcus (20%) . The lethality was 18% . About three fourths of the children dying from the infection, died during the first week of life . For the children who did not die during their stay in hospital, the duration of hospitalization was longer than 4 weeks in 41%.

Nephron, 1995, 71(2), 133 - 7
Impaired polymorphonuclear neutrophil function in end-stage renal failure and its correction by continuous ambulatory peritoneal dialysis; Porter CJ et al.; The polymorphonuclear neutrophil (PMN) function of 22 patients with end-stage renal failure (ESRF) was studied immediately before and 3 months after starting continuous ambulatory peritoneal dialysis (CAPD) and compared with a control group of healthy normal volunteers . The PMN functions studied were phagocytosis and killing of Staphylococcus epidermidis and respiratory burst activity . The results show that in the presence of normal pooled human serum PMN from patients with ESRF before CAPD treatment phagocytose bacteria normally, but have impaired killing . Before treatment, the PMN from patients with ESRF also showed an increase in both unstimulated and stimulated superoxide anion production . Abnormal PMN function was corrected by CAPD treatment, suggesting the involvement of a dialyzable toxin.

Microbiol Immunol, 1995, 39(6), 369 - 77
Optimum culture conditions for production of exfoliative toxin by Staphylococcus hyicus; Watanabe T et al.; Optimum culture conditions for the production of exfoliative toxin by Staphylococcus hyicus (shET) were examined . High shET activity was obtained from the culture filtrate of HI and TY broth inoculated with S . hyicus . The pH in these two media ranged from 7 to 8.5 during bacterial culture, while the lowest pH in TS and BHI broth was less than 6 . shET activity in the culture filtrate from TY broth inoculated with 10(7) CFU of S . hyicus per ml was higher than that in TY broth inoculated with 10(6) and 10(8) CFU of bacteria per ml . When shET activity in the culture filtrate was measured under various shaking conditions, the culture filtrate shaken at 75 oscillations per min had the highest shET activity of the five shaking conditions . shET activity of the culture filtrate of TY broth to which protease inhibitor had been added was the same as that of TY broth without inhibitor . shET activity in a shaking culture in an Erlenmeyer flask was also the same as that in sac culture and that in shaking culture using a shaking (Sakaguchi) flask . shET activity in TY broth supplemented with 100 mM glucose was significantly lower than that in TY broth without glucose . Based on the above results, the optimum culture conditions for the production of shET were as follows: inoculation of 3 x 10(9) CFU of S . hyicus strain P-1 into 300 ml of TY broth in a 2,000-ml Erlenmeyer flask, and incubation at 37 C with shaking at 75 oscillations per min.(ABSTRACT TRUNCATED AT 250 WORDS)

Adv Perit Dial, 1995, 11, 182 - 6
The efficacy of intraperitoneally administered gentamicin and rifampin as initial treatment of peritoneal dialysis-related peritonitis; Brulez HF et al.; For the initial treatment of peritonitis complicating peritoneal dialysis (PD), we use intraperitoneally administered gentamicin (broad spectrum and low costs) and rifampin (intracellular bactericidal activity) . In order to assess the efficacy of this treatment, the outcome of 248 suspected episodes of peritonitis (abdominal pain, cloudy effluent, and a leukocyte count over 100/mm3) was evaluated . Of 227 cases with a positive culture of the PD effluent, one bacterial species was cultured in 188 cases (75.8%), more than one in 32 cases (12.9%), and in 7 cases (2.8%) yeasts . In 87.2% of the culture-positive cases, a good clinical response to the initialized antibiotic therapy was found . In 20 cases (8.1%) antibiotic treatment was discontinued within one week because no micro-organisms were cultured . In one case no effluent was cultured . Although in vitro resistance or indifference to both antibiotics was found in 45 cases (19.8%), in only 29 culture-positive cases (12.8%) the clinical condition did not improve on initial therapy . Of the peritonitis episodes in which micro-organisms resistant to both antibiotics were cultured, 23 were Staphylococcus epidermidis, 5 were E . coli, 7 were yeasts, and there were miscellaneous (mostly enteral) bacteria in 10 cases . In the studied period no significant changes were found in the susceptibility of the cultured microorganisms to gentamicin and rifampin . Susceptibility profile per episode, however, showed an increasing resistance against both antibiotics . It is concluded that the combination of gentamicin and rifampin as initial treatment of peritonitis is effective in most (87%) cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Adv Perit Dial, 1995, 11, 176 - 8
Treatment of recurrent and resistant CAPD peritonitis by temporary withdrawal of peritoneal dialysis without removal of the catheter; Locatelli A et al.; Recurrent and resistant continuous ambulatory peritoneal dialysis (CAPD) peritonitis is usually treated by removal of the catheter and temporary hemodialysis . We treated 3 patients: 1 with resistant Klebsiella peritonitis and 2 with recurrent peritonitis (one due to Staphylococcus and the other to Enterococcus), by stopping CAPD for a 2-4 week period, leaving the catheter in situ and continuing antibiotic therapy . All 3 patients had resolution of their infections and restarted CAPD . This therapeutic modality reduced catheter replacements, limited admissions to the hospital, reduced psychological impact, and diminished risks and costs of CAPD.

Adv Perit Dial, 1995, 11, 134 - 8
Role of automated peritoneal dialysis within a peritoneal dialysis program; Viglino G et al.; We wished to assess the impact of automated peritoneal dialysis (APD) on the peritoneal dialysis (PD) program . From November 1981 to December 1993, 112 patients were started on hemodialysis (HD) as first treatment and 88 on PD {continuous ambulatory peritoneal dialysis (CAPD): 78, APD: 10}; respective average ages were 61 +/- 14 and 62 +/- 13 years . To December 1985, APD was used as first treatment of PD in 1/29 patients (3.4%), while subsequently, on the basis of a clinical and social-aptitude assessment protocol, it was used in 9/59 patients (15.2%) with PD indication and CAPD contraindications (work: 2 patients, partner required: 7 patients) . Of the patients who interrupted CAPD, APD was used in 9/21 patients (reason: social aptitude, 28.6%; clinical, 71.4%) . Technique survival after 5 years proved no different in HD versus PD (87% vs 82%, p = NS), whereas in HD versus CAPD it was different (87% vs 62%, p < 0.025) . The incidence of peritonitis in APD and CAPD with the Y-set was comparable (1/37 vs 1/40 episode/patient-months), while germ distribution was different (p < 0.001) with Staphylococcus epidermidis prevailing in APD (59%) . Based on our experience, APD may extend method acceptance criteria and reduce the technique dropout rate in PD; however, connection technique may need to be improved in order to reduce the risk of peritonitis from exogenous contamination.

Przegl Lek, 1995, 52(8), 375 - 81
{Circulating immune complexes in airborne allergy--a protective role?}; Jarzab J et al.; Circulating immune complexes (KI) were analyzed in 78 patients with airborne allergy and compared to 34 persons of control group . KI were isolated by precipitation with 3% polyethylene glycol . KI-IgG, KI-IgA, KI-IgM were measured by radial immunodiffusion, KI-IgE by an immunofluorometric assay . Staphylococcal Protein A (SpA) binding to KI was measured by ria . Relations between the level of various KI and the diagnosis of allergic disease, the kind of allergen (related also to the exposition) and the symptoms of atopy were evaluated . Serum levels of KI-IgE, KI-SpA, and KI-IgM were distinctly elevated in airborne allergy, with the positive correlation between KI-SpA and KI-IgM . KI-IgE were related to the kind of sensitizing allergen and to the exposition to the pollen allergens . Most distinct differences in KI level were observed not in relation to the kind of disease or of the sensitizing allergen, but in relation to the symptoms and intensiveness of atopy . Patients showing a greater number and intensiveness of symptoms exhibited higher levels of KI-SpA . However, one of the symptoms scored (dyspnea) was connected with the decreased level of KI-SpA in these patients . We interpret the results in favour of the hypothesis of the dual role of KI in airborne allergy . We propose a protective role for KI-Spa, which contain IgG and may bind IgE, thus protecting the target organs of allergy.

Med Sci Sports Exerc, 1995 Jan, 27(1), 73 - 8
Immunomodulatory effects of acute exercise bout in sedentary and trained rats; Lin YS et al.; The immune response to acute exercise after long-term training was evaluated in rats . After 10 wk of exercise training on a drum exerciser, both sedentary control and trained groups ran with the intensity of 70% of peak oxygen consumption (VO2peak) for 10 min immediately before sacrifice . The mitogenic activity of spleen lymphocytes in trained group to staphylococcal enterotoxin B, a polyclonal T cell activator, decreased as compared to the sedentary control following acute exercise . However, proliferative response to lipopolysaccharide, a B cell mitogen, was augmented . Furthermore, the interleukin-2 production was reduced in the trained group . The lymphocyte subpopulations in the spleen and the peripheral blood lymphocytes were analyzed by flow cytometry . Although the tendency of changes in some populations was observed in trained vs untrained groups, no statistically significant difference was manifested . The serum levels of both norepinephrine and epinephrine increased immediately after acute exercise . The increase in serum lactate concentration was observed following acute exercise in sedentary control, but less prominent in the trained group.

Comp Immunol Microbiol Infect Dis, 1995 Jan, 18(1), 9 - 16
Intestinal immune response induced in mice by staphylococcal enterotoxin B; Bhatti AR et al.; To investigate the induction of intestinal immunity to staphylococcal enterotoxins (SE) we have chosen the mouse as an experimental model . Since this species is devoid of emetic mechanism, SE can be administered orally without any loss . Mice were treated orally and/or parenterally with staphylococcal enterotoxin B (SEB) . The anti-SEB response, either in serum or in the supernatant of in vitro cultured intestinal fragments was determined by enzyme immunoassay (EIA) . The results showed that orally given SEB induced specific antibodies both in serum and intestinal secretions . Compared to oral route alone, parenteral followed by oral administration of SEB induced a higher intestinal response with IgA as predominant isotype . Although these results cannot directly be extrapolated to humans or animals with emetic reaction to SE, they do show the implication of intestinal immune system in response to this group of toxins.

Am J Nephrol, 1995, 15(1), 78 - 81
Metastatic endophthalmitis in dialysis patients; Smith KG et al.; Metastatic bacterial endophthalmitis is a rare but devastating complication of septicaemia . We present 3 recent cases occurring in dialysis patients . Septicaemic dialysis patients seem particularly prone to staphylococcal endophthalmitis which developed in the face of therapeutic serum concentrations of appropriate antibiotics in all 3 cases . Prompt treatment, including consideration of intravitreal antibiotics, is required if vision is to be preserved . Early signs of endophthalmitis should be actively sought in septicaemic dialysis patients, as they are easily overlooked.

Eur J Immunol, 1995 Jan, 25(1), 92 - 101
Cross-reactive trinitrophenylated peptides as antigens for class II major histocompatibility complex-restricted T cells and inducers of contact sensitivity in mice . Limited T cell receptor repertoire; Kohler J et al.; The induction of contact sensitivity in mice by hapten reagents such as trinitrochlorobenzene (TNCB) involves the activation of class II major histocompatibility complex (MHC)-restricted, hapten-specific, CD4+ T cells . Reports from different laboratories have indicated that the relevant antigenic epitopes in such reactions might include hapten-conjugated, MHC class II-associated peptides . This study for the first time directly demonstrates that hapten-peptides account for the majority of determinants recognized by trinitrophenyl (TNP)-specific CD4+ T lymphocytes . The sequences of those TNP carrier peptides do not have to be related to mouse proteins . Thus, we show that TNP-modified peptides derived from mouse IgG, pigeon cytochrome c or staphylococcal nuclease known to bind to I-Ab or from lambda repressor with specificity to I-Ad as well as TNP-proteins such as bovine serum albumin, ovalbumin or keyhole limpet hemocyanin all create class II-restricted hapten determinants for a number of TNP-specific T cell clones and hybridomas . All of these cells were induced with cells modified by trinitrobenzene sulfonic acid (TNBS) . In addition, we present arguments indicating that individual TNP-specific helper T cells may cross-react with different TNP-peptides bound to identical class II molecules . Chemical treatment of antigen-presenting cells with TNCB or TNBS may thus result in a limited number of particularly repetitive immunodominant hapten epitopes . Immunodominant epitopes were also indicated by an overrepresentation of the TCR elements V beta 2 and V alpha 10 in I-Ab/TNP-specific T cells . Most importantly, however, we demonstrate that TNP attached to lysine 97 in the staphylococcal nuclease peptide 93-105 (i.e . a clearly "non-self" sequence) is able to prime mice for subsequent elicitation of contact sensitivity by TNCB in the absence of foreign protein . We take this to indicate that those TNP-peptide determinants defined by us as immuno-dominant are responsible for the induction of contact sensitivity to haptens.

Eur J Immunol, 1995 Jan, 25(1), 75 - 9
Distinct splicing of CD45 mRNA in activated rat gamma delta cytotoxic T lymphocytes; Hansson J et al.; Previous studies have demonstrated tumor- and allo-specific cytotoxic gamma delta T lymphocytes in rats . In this report we define the surface phenotype of these T cell receptor (TCR) gamma delta+ T cells and demonstrate distinct CD45 mRNA splicing in activated gamma delta cytotoxic T lymphocytes (CTL) . gamma delta T lymphocytes in the blood and the peritoneal cavity were TCR alpha beta-CD3+CD8 alpha+CD45RC+ but expressed variable levels of LFA-1 molecules . Normal peritoneal gamma delta T lymphocytes, peritoneal gamma delta T cells from rats injected with the bacterial superantigen staphylococcal enterotoxin A (SEA) as well as gamma delta T lymphocytes in peripheral blood were all LFA-1low . Peritoneal gamma delta T cells from tumor-, and allo-sensitized rats were either LFA-1low or LFA-1high and specific cytotoxicity was highly enriched in the LFA-1high subset . No cytolytic activity against SEA-presenting cells was recorded in gamma delta T cells from SEA-injected rats . Different isoforms of CD45 in T cells are generated by alternative mRNA splicing of exons 4, 5, 6 (or A, B and C, respectively) and the recently described alternate exon 7 . CD45 splicing in sorted gamma delta T cells was evaluated utilizing reverse transcription polymerase chain reaction . Normal peritoneal gamma delta T cells expressed exon(578), exon(678), exon(78) and the extensively spliced exon(8) variant . Peritoneal gamma delta T cells from rats sensitized with irradiated syngeneic tumor cells, allogeneic cells or bacterial superantigen SEA as well as gamma delta T lymphocytes in peripheral blood contained the full-length exon(45678), as well as the exon(5678), exon(578), exon(678) and exon(78) splicing products . Notably, the exon(8) variant was also seen in peritoneal gamma delta T cells of SEA-sensitized rats . Sorted tumor-specific LFA-1high gamma delta CTL expressed exon(45678), exon(5678), exon(578), exon(678) and exon(78) CD45 splicing products whereas the non-cytolytic LFA-1low gamma delta T cell subset also contained exon(8) variant . In summary, it is concluded that antigen-specific TCR gamma delta+ CTL express high levels of LFA-1 and that the splicing machinery in these cytolytic cells favors expression of the exon(45678) and exon(5678) CD45 splicing products whereas the exon(8) variant is lost . TCR alpha beta+ CTL express high levels of LFA-1 but are devoid of the full-length exon(45678) splicing product . The different CD45 splicing patterns found in alpha beta CTL and gamma delta CTL indicate different molecular requirements in respect to CD45 during activation and differentiation of these T lymphocyte subsets.

Cell Immunol, 1995 Jan, 160(1), 58 - 64
Staphylococcal enterotoxin B modulates V beta 8+ TcR-associated T-cell memory against conventional antigen; Bell SJ et al.; Primary in vivo challenge with the superantigen staphylococcal enterotoxin B (SEB) induces polyclonal proliferation of an unusually large proportion of circulating T-cells that bear the V beta 8-T-cell receptor (TcR) domain . Early and vigorous proliferation of V beta 8+ T-cells precedes their selective deletion, leaving the host unresponsive upon rechallenge with the native immunogen SEB . Nonetheless, this induction of anergy is incompletely understood . Recently we demonstrated that more cells than just V beta 8+ T-cells undergo clonal proliferation after challenge with SEB (Cell . Immunol . 154, 440, 1994) . These findings suggested that non-V beta 8+ T-cells may have a role in the induction of superantigen-induced anergy . To further investigate this, we enumerated CD4+ and CD8+ T-cells in lymph nodes and spleens from Balb/c mice at various times after primary and secondary challenge with either a high or a low dose of SEB . Using these kinetic data we investigated whether challenge with SEB would modulate antigen-specific V beta 8-associated T-memory responses . To this end, the V beta 8+ T-cell-associated responses induced by SEB were compared with the V beta 8+ TcR-associated memory responses induced by the nominal antigen sperm whale myoglobin (SWM) . Results indicated that challenge of SWM-primed mice with SEB abrogated the V beta 8-associated SWM-specific T-cell memory for an extended but transient period of time . Moreover, prechallenge with SEB blocked the establishment of de novo V beta + T-cell-mediated immunity . These findings suggest that administration of low and controlled doses of microbial superantigen could provide long-term suppression of antigen-specific cell-mediated immunity.

South Med J, 1995 Jan, 88(1), 47 - 51
Clinical experience with 180 inflatable penile prostheses; Randrup ER; I present my experience with 180 inflatable penile prostheses inserted in 167 patients with organic sexual impotence of 4 months to 30 years in duration . Mean age was 59.3 years . Prostheses used were AMS 700 (9), Flexiflate (14), Mentor Mark II (99), and Mentor Alpha I (58) . Mean follow-up was 43.5 months . Overall complication rate was 17.8% (32 cases) . In 24 cases, the complications were major (infection, malfunction, and extrusion) and necessitated reoperation . The remaining complications (8 cases) were minor (partial deviation, autoinflation), and surgical intervention was not needed . Staphylococcus epidermidis was the offending microorganism in 7 of 9 infections . In general, my results with these penile prosthesis implants suggest that they are a valid form of treatment for organic impotence.

Arch Pathol Lab Med, 1995 Jan, 119(1), 59 - 63
The neuropathology of cardiac allograft transplantation . An autopsy series of 18 patients; Prayson RA et al.; OBJECTIVE--To examine the neuropathologic findings associated with cardiac transplantation and to assess the role, if any, that the neuropathology had in the patient's death . DESIGN--Retrospective autopsy series of 18 patients . SETTING--Tertiary referral center with a high volume of cardiac transplantation . PATIENTS--Cardiac transplant patients who died and in whom a complete autopsy was performed . RESULTS--Eighteen patients, including 11 men and seven women, comprised the study group . Patients ranged in age from 27 to 59 years (mean 48 years) . Postoperative survival ranged from 1 to 1517 days (mean 334 days) . Autopsies in 15 (83%) patients showed neuropathology; in only one patient, however, was cause of death attributed to these findings . The most common neuropathology was related to anoxia or infarction: ischemia or focal neuronal necrosis (n = 9), diffuse anoxic encephalopathy (n = 3), infarction (n = 6) . Other findings included infection with Nocardia, Aspergillus, and Staphylococcus (n = 3) and hemorrhage (n = 3) . Of the seven patients who received transplants for ischemic heart disease, five had evidence of cerebral ischemia or infarction . Systemic pathology encountered included infection in 13 patients, ischemia and/or infarction in 11 patients, and hemorrhage in six patients . Cause of death was attributed to infection in nine patients, cardiac rejection in five patients, disseminated intravascular coagulation/hemorrhage in three patients, and systemic amyloidosis in one patient . CONCLUSIONS--The most common neuropathologic findings are related to ischemia and infarction . Neuropathologic findings were only rarely the main cause of death, but they were a significant cause of morbidity in patients who have undergone cardiac allograft transplantation.

J Infect Dis, 1995 Jan, 171(1), 220 - 4
Potentiation of Staphylococcus epidermidis catheter-related bacteremia by lipid infusions; Shiro H et al.; Lipid infusions given to low-birth-weight neonates are associated with coagulase-negative staphylococcal bacteremia . A rabbit model of catheter-related Staphylococcus epidermidis bacteremia was used to compare infection rates between animals receiving 20% lipid infusions with those receiving 10% glucose infusions . Bacterial growth was detected in 43 (66%) of 65 blood cultures obtained during lipid infusions but only 6 (7%) of 91 blood cultures obtained during glucose infusions (P < .001) . S . epidermidis was cultured from 7 of 9 catheters recovered from lipid-infused rabbits but from only 3 of 13 catheters from glucose-infused rabbits (P = .016) . Lipid but not glucose solutions containing low levels of protein (0.1%-1.0%) supported the survival and growth of S . epidermidis . These results suggest that lipids contribute to the survival and growth of S . epidermidis on contaminated catheters, enhancing the potential of these organisms to disseminate and cause bacteremia.

Med Hypotheses, 1995 Jan, 44(1), 20 - 7
Preliminary evidence for idiotype-antiidiotype immune complexes cross-reactive with lymphocyte antigens in AIDS and lupus; Root-Bernstein RS; Several investigators have proposed that autoimmunity may be induced by idiotype-antiidiotype antibody networks . It is generally assumed that the antiidiotype is produced in response to the idiotype, and therefore that autoimmune diseases have single antigenic initiators . The theory of multiple-antigen-mediated autoimmunity (MAMA) proposes, on the other hand, that idiotype and antiidiotype result from two primary immune responses to two chemically complementary antigens . Because of the complementarity of the antigens, and the complementarity of the antibodies for the antigens, the antibodies will themselves be complementary . They will thus form circulating immune complexes, the self-nonself distinction diffusion (DAD) experiments (a modification of Ouchterlony immunodiffusion), in which 1800 pairs of antibodies were screened for their ability to form precipitating complexes . Four sets of antibodies associated with AIDS (HIV + Staphylococcus; HIV + Mycoplasma; CMV + Mycoplasma; and HBV + Mycoplasma) specifically precipitated each other, and one of the antibodies in each set also precipitated monoclonal antibodies against one or more lymphocyte protein markers . These results therefore demonstrate that idiotype-antiidiotype antibodies can be elicited by independent antigens and may induce AIDS-related forms of autoimmunity directed at lymphocytes.

Curr Microbiol, 1995 Jan, 30(1), 11 - 6
Purification and characterization of extracellular Staphylococcus warneri lipase; Talon R et al.; The extracellular lipase of Staphylococcus warneri was secreted as a protein with an apparent molecular mass of 90 kDa . It was then sequentially processed in the supernatant to a protein of 45 kDa . Tryptic digestion of the crude extract resulted in a homogeneous sample containing only the 45-kDa form . Purification was achieved by hydrophobic chromatography . Purified lipase had an optimum pH of 9.0 and an optimum temperature of 25 degrees C . The enzyme was stable within the range pH 5.0-9.0; it had a broad substrate specificity . The results of inhibition studies were consistent with the view that lipases possess a serine residue at the catalytic site.

Lett Appl Microbiol, 1995 Jan, 20(1), 19 - 24
Comparative studies of the growth of Staphylococcus carnosus with or without glucose; Arkoudelos JS et al.; Lactic acid and acetic acid are metabolic products of Staphylococcus carnosus irrespective of the media used . Lactate was produced at the expense of glucose (when available) while acetic acid was produced at the expense of lactate under aerobic or semi-aerobic conditions . The production of acetic acid could affect seriously the flavour and aroma of fermented sausages.

Arch Pediatr, 1995 Jan, 2(1), 47 - 51
{Favorable outcome of orbital nasal sinus mucormycosis complicating the induction treatment of acute lymphoblastic leukemia}; Gebhard F et al.; BACKGROUND--Most cases of mucormycosis occur in immunosuppressed children . Intracranial extension is lethal and must be prevented with early specific treatment . CASE REPORT--A 42 month-old boy was admitted suffering from acute lymphoblastic leukemia . Edema of the left eyelid developed on the sixth day of induction chemotherapy . Mucormycosis was suspected because of gradual extension of infection to nasal ala and periorbital area with fever, edema of nasal turbinates and nasal black secretions . Chemotherapy was discontinued and the patient was given intravenous amphotericin B (1.0 mg/kg/day) and heparin associated with G.CSF . Improvement was only temporary and scan examination performed on day 17 showed involvement of the orbit, eye and wall of the maxillary sinus; cultures of secretions were positive for staphylococcus and Absidia corymbifera . Remission of leukemia was obtained a few days later permitting surgical resection of involved tissues on day 30 . A relapse of mucormycosis was observed six weeks later despite prolonged administration of amphotericin B requiring extended resection of necrotic areas and replacement of amphotericin B by its liposomal form (Ambisome) . Bone marrow relapse of leukemia required further chemotherapy . The patient is in good condition 30 months after the initial symptoms . CONCLUSION--Our patient seems to be the first with prolonged remission of facial mucormycosis and acute leukemia despite relapse of both diseases . This favorable outcome could be due to the use of Ambisome.

Clin Infect Dis, 1995 Jan, 20(1), 24 - 9
Persistence of a multiresistant clone of Staphylococcus epidermidis in a neonatal intensive-care unit for a four-year period; Lyytikainen O et al.; A cluster of cases of Staphylococcus epidermidis bacteremia in a neonatal intensive-care unit (NICU) during 1991 raised the question of whether these infections were caused by a single strain . Sixty-seven isolates of S . epidermidis from blood cultures of 56 neonates treated in the NICU between 1986 and 1992 and 54 control strains from other patients with bacteremia were characterized by three typing methods: antibiogram, plasmid profile, and ribotype . Plasmid profiles and ribotype patterns indicated that 11 (16%) of the 67 episodes of S . epidermidis bacteremia in the NICU were caused by a single strain . Although this epidemic strain did not account entirely for the increase in the incidence of bacteremia in the NICU, it did persist for 4 years during the study period . Other clones responsible for smaller outbreaks were also found . These results suggest that S . epidermidis cross-infections are very common in the NICU setting.

J Burn Care Rehabil, 1995 Jan-Feb, 16(1), 62 - 4
The burn unit as a resource for the management of acute nonburn conditions in children; Sheridan RL et al.; Burn units bring together resources to manage large complex wounds, organ failures, and the hypermetabolic response to injury . These resources can also facilitate management of other problems such as purpura fulminans, toxic epidermal necrolysis, staphylococcal scalded skin syndrome, and major mechanical soft-tissue injuries . During a recent 10-year interval 2.4% of all acute admissions to a regional pediatric burn facility were in this category and form the basis for this review.

Int Immunol, 1995 Jan, 7(1), 37 - 43
Impaired CD28-mediated co-stimulation in anergic T cells; Suzuki G et al.; We have investigated a CD28 co-stimulation in anergic T cells in staphylococcal enterotoxin B-inoculated mice by stimulating the cells with a plate-coated anti-TCR antibody in the presence or absence of an anti-CD28 antibody . CD28 co-stimulation increased the levels of IL-2 and IL-4 mRNAs in naive CD4+V beta 8+ T cells . However, it did not increase the levels of IL-4 mRNA at all and only partially increased those of IL-2 mRNA in anergic T cells . It was demonstrated that CD28 co-stimulation was impaired so that it no longer stabilized cytokine mRNAs in anergic cells . The levels of IL-4 mRNA in response to TCR stimulation were higher in anergic T cells than those in naive T cells in spite of the defective CD28 co-stimulation in the former cells . Anergy induction and generation of a Th2-type immune response in vivo are discussed.

Int Immunol, 1995 Jan, 7(1), 105 - 14
Superantigen-reactive T cells that display an anergic phenotype in vitro appear functional in vivo; Heeg K et al.; Clonal deletion and/or inactivation establishes tolerance to self antigens . Endogenous and exogenous (bacterial) superantigens, like the staphylococcal enterotoxins, induce ligand-specific clonal anergy in vivo and thus are believed to mirror aspects of post-thymic tolerance mechanisms in mature peripheral T cells . Here we analyzed the level of anergy of ligand-responsive V beta 8+ T cells from staphylococcal enterotoxin B (SEB)-primed mice in vivo and in vitro . Upon in vitro restimulation with SEB, CD4+V beta 8+ and CD8+V beta 8+ T cells failed to produce IL-2 . However, functional IL-2 receptors were triggered, since supplementation with IL-2 induced clonal growth in virtually all CD4+V beta 8+ and CD8+V beta 8+ T cells as determined by limiting dilution analyses . Thus in vitro unresponsiveness of lymphocytes from SEB-primed mice reflects the inability of SEB-reactive V beta 8+ T cells to produce IL-2 . Surprisingly, anergy as defined in vitro was at variance with that in vivo . Following further challenge with SEB, systemic and acute lymphokine production (including IL-2 and tumor necrosis factor) occurred with almost identical peak values and kinetics to primary in vivo responses, and D-galactosamine-sensitized mice succumbed to lethal shock . Polymerase chain reaction analyses revealed that CD4+V beta 8+ expressed IL-2-specific mRNA in vivo upon restimulation with SEB . While lymphokine production and expression of the IL-2 receptor was similar to the response to in vivo primary stimulation, only CD8+V beta 8+ T cells expanded clonally upon reintroduction of SEB in vivo . Hence primed V beta 8+ T cells challenged with SEB display in vitro anergy yet in vivo responsiveness, at least in part . We conclude that the state of anergy is reversible, dependent upon the quality of activation signals provided in in vivo rather than in in vitro culture conditions.

Arch Toxicol, 1995, 69(3), 149 - 59
Characterization of the tobacco glycoprotein surface binding property of heart and skeletal muscle cells . I . Modulation of the heart cell membrane TGP interaction by anti-TGP IgG; Santos-Buch CA et al.; Monolayers of L6 rat skeletal myoblast cells formed surface binding isotherms with the purified tobacco leaf glycoprotein TGP1 and the enriched cigarette tar glycoprotein TGP2 . Scatchard analysis showed that the binding in the range of the limited concentrations tested was to a single class molecule and the calculated affinity constant (Kd) for TGP1 and TGP2 showed similar values (9.78 x 10(-13) M and 3.09 x 10(-13) M, respectively) . The bound TGPs were almost totally displaced by excess nonradiolabeled molecules . The calculated Bmax of the L6 myoblast monolayer was 2.93 fmol for TGP1 and 0.217 fmol for TGP2 per 32.2 mm2 . Guinea pig heart sarcolemma binding isotherms were also formed with radiolabeled TGP1 and TGP2 . The interaction of tobacco leaf TGP1 with the heart cell membranes was irreversible because only 15-20% of the bound TGP1 was displaced by 100-fold, non-labeled molecules but the interaction of tar TGP2 with heart sarcolemma was reversible and probably saturable . The heart sarcolemma TGP2 affinity constant (Kd) was 5.88 x 10(-7) M and the Bmax, 2.45 x 10(-8) M per 12.5 micrograms sarcolemma . Pretreatment of heart sarcolemma with increasing concentrations of leaf TGP1 did not displace tar TGP2 binding but its absorption on the membrane resulted in increased TGP2 sarcolemma attachment by a complex and unexplained mechanism . Increasing concentrations of the sera of 10 of 15 guinea pigs (67%) that received mainstream emissions of tobacco smoke from a University of Kentucky cigarette smoking machine for 152 days, displaced cigarette tar TGP2 heart cell sarcolemma attachment and this inhibition was significantly different from that produced by the sera of sham smoked and of non-exposed animals (Mann-Whitney test, p = 0.0082) . Staphylococcus protein A inhibited the displacement of TGP2 produced by the sera of cigarette smoke exposed guinea pigs and this observation indicated that this action was mediated by IgG molecules . The specific immunoprecipitation of a radiolabeled surface epitope of the L6 myoblast monolayers pretreated with TGP1 or TGP2 by immune IgG against TGP2 and by the IgG of an antiserum against standard TGP showed that the tobacco glycoproteins attached to a unit polypeptide of the plasma membrane of the muscle cells of approximately 76 kDa . These data support the notion that TGP molecules in cigarette smoke are absorbed systemically on smoking and may have a direct toxic effect when they attach to the surface TGP binding proteins of heart and skeletal muscle cells.

J Clin Lab Anal, 1995, 9(2), 81 - 8
Contribution of the host to test results in assays of Staphylococcus epidermidis; Herndon B et al.; From two human populations (one pediatric and one adult), clinically diagnosed with Staphylococcus epidermidis (S . epidermidis) sepsis of similar severity, bacteria were isolated from pre-antibiotic blood samples and evaluated for virulence . The LD50 of the bacteria in a mouse model was performed, with evaluation of animals dying acutely following intravenous S . epidermidis administration . More simple assays of virulence were also performed, including bacterial adherence to a fibrin clot and carbohydrate specific lectin binding . The eight pediatric-host S . epidermidis isolates required a significantly larger dose to produce lethality in dosed animals (LD50) when compared to the 20 adult-host S . epidermidis isolates . The fibrin clot assay, a test that has corroborated bacterial virulence in endocarditis models, did not differentiate the groups: all but one of the 28 isolates were well above the adherence seen with the ATCC control, suggesting endocarditis-producing potential . Glycocalyx (slime) from eight of the more virulent isolates showed reactivity with a glucose-specific biotinylated lectin which was lacking in other isolates . Necropsy of mice dying at 12 hr showed S . epidermidis strain differences in specific organ effects . Overall, this study demonstrates the utility of the LD50 to provide a highly sensitive quantification of bacterial virulence . Necropsy of test animals dying acutely has showed an apparent organ tropism of some of these isolates which are usually considered harmless commensals.

Eye, 1995, 9 ( Pt 1), 116 - 8
Bacterial contamination of nylon corneal sutures; Heaven CJ et al.; We report the findings of a prospective study into the bacterial contamination of monofilament nylon sutures removed from corneal wounds following cataract surgery . Sutures were classified as tight, loose or broken at the time of removal . Loose and broken sutures showed significantly more bacterial contamination than tight sutures (p < 0.001, chi squared) . Positive cultures were obtained from 2 (6.2%) of 32 tight sutures, 14 (38.9%) of 36 loose sutures and 11 (37.9%) of 29 broken sutures . Staphylococcus epidermidis was the most commonly isolated organism (isolated in pure growth from 22 (81.5%) of 27 positive cultures) . These findings may explain the occasional association of biodegraded corneal monofilament nylon sutures and suppurative keratitis and highlight the potential risk of seeding a suture track infection at the time of suture removal . They also emphasise the need for prophylactic topical antibiotic when removing biodegraded sutures.

Bioconjug Chem, 1995 Jan-Feb, 6(1), 93 - 100
Strand invasion by oligonucleotide--nuclease conjugates; Corey DR et al.; Conjugates consisting of staphylococcal nuclease crosslinked to oligonucleotides hybridize to supercoiled duplex DNA by Watson--Crick base-pairing . Here we describe this strand invasion . Affinity cleavage by these conjugates provides a probe for the local topology of the DNA duplex and is most efficient at a target DNA sequence known to form a cruciform . Additional supercoiling of the substrate DNA increases selective cleavage at other sequences . Hybridization of the conjugate to duplex DNA is temperature dependent and is stable over time . Affinity cleavage is not substantially inhibited by a 200-fold excess of the analogous unmodified oligonucleotide, demonstrating that hybridization of the unmodified oligonucleotide must be less favored and that the nuclease is involved in substrate binding . Surprisingly, affinity cleavage is also not effectively inhibited by complementary oligonucleotides unless they contain an extended 5'-sequence capable of separate interactions with the nuclease domain of the conjugate . These results suggest that the oligonucleotide-nuclease conjugate prefers to hybridize to target sequences which will allow interactions with both the oligonucleotide and the nuclease domains . Affinity cleavage by oligonucleotide-nuclease conjugates provides general insights for the design of oligonucleotides and their conjugates for strand invasion and affords a convenient competition assay for their hybridization.

Annu Rev Biophys Biomol Struct, 1995, 24, 435 - 62
Site-directed mutagenesis with an expanded genetic code; Mendel D et al.; A biosynthetic method has been developed that makes possible the site-specific incorporation of a large number of amino acids and analogues within proteins . In this approach, an amber suppressor tRNA chemically aminoacylated with the desired amino acid incorporates this amino acid site specifically into a protein in response to an amber codon introduced at the corresponding position in the protein's DNA sequence . Using this method, precise changes within a protein can be made to address detailed structure-function questions . A series of fluorinated tyrosine analogues and linear, branched, and cyclic hydrophobic amino acids have been used to determine the impact of hydrogen bonding and hydrophobic packing, respectively, on protein stability . Glutamate analogues and conformationally restricted amino acids have been used to probe the mechanisms of staphylococcal nuclease and ras . In addition, this technique has been used to construct photocaged proteins and proteins containing photoaffinity labels, spin labels, and isotopic labels at specific positions in the protein sequence suitable for biophysical studies.

Annu Rev Biophys Biomol Struct, 1995, 24, 141 - 65
Thermodynamics of partly folded intermediates in proteins; Freire E; Until recently, the energetics of protein-folding intermediates eluded direct measurement by high-sensitivity microcalorimetric techniques . But during the past year, the direct measurement of thermodynamic parameters for folding intermediates of alpha-lactalbumin, apomyoglobin, cytochrome c, and staphylococcal nuclease has provided new insights on the nature of the forces involved in the stabilization of nascent protein structures . In this review, I summarize those results and discuss the structural implications of the observed thermodynamic behavior.

J Interferon Cytokine Res, 1995 Jan, 15(1), 39 - 45
Type I interferon inhibition of superantigen stimulation: implications for treatment of superantigen-associated disease; Soos JM et al.; The interferons (IFNs) are a family of secretory glycoproteins possessing potent antiviral, antiproliferative, antimicrobial, and immunomodulatory activities . It has been shown that the IFNs and superantigens have an important effect on the course of certain autoimmune disorders, and thus we have examined the effect of the type I and type II IFNs on superantigen-induced stimulation . The type I IFNs, alpha, beta, and tau, inhibited induction of T cell proliferation by several staphylococcal enterotoxin superantigens; the type II IFN, gamma, was without effect . The type I IFNs inhibited T cell proliferation to the same extent, approximately 50% at 10(3) units of IFN/ml, and in a dose-dependent manner . Consistent with inhibition of proliferation, the type I IFNs also inhibited IL-2 production as well as levels of IL-2 receptor expression . Inhibition was not increased by using the IFNs in combination, suggesting that they inhibited proliferation by the same mechanism . IFNs alpha and beta, but not IFN-tau, were toxic to cells at high concentrations (> or = 10(4) units/ml) . Thus, the mechanism by which type I IFNs inhibit cell proliferation differs from that associated with their toxic effects . A partial reduction of V beta-specific superantigen-induced T cell expansion by type I IFNs was also demonstrated using flow cytometry . We recently showed that superantigens play an important role in the reactivation of experimental allergic encephalomyelitis . The potent antiproliferative activities of the type I IFNs strongly suggest the further study of their use as therapies for superantigen-associated diseases, such as multiple sclerosis and other autoimmune disorders, as well as toxic shock syndrome.

Biochimie, 1995, 77(3), 217 - 24
Identity of macromolecules present in the extracellular slime layer of Staphylococcus epidermidis; Karamanos NK et al.; The extracellular slime layer of Staphylococcus epidermidis ATCC 35983 contains: a) two non-anionic carbohydrate containing proteins degradable with papain of molecular masses 250 and 125 kDa; b) a polydisperse but homogeneously-charged acidic population with M(r) ranging from 120,000 to 35,000 (average M(r) (80,000) containing a polysaccharide covalently bound to a small peptide; c) a papain degradable macromolecule (molecular mass 60 kDa) bearing acidic carbohydrates covalently bound to protein; and c) two acidic polysaccharides strongly retained by the anion-exchange column; one polysaccharide is sulphated and has a molecular mass of 20 kDa; the other has a higher charge density and a molecular mass of 12.5 kDa . The results obtained clearly demonstrate the presence of discrete macromolecules in the extracellular material of slime-producing S epidermidis, the majority of which contain acidic carbohydrates, whose biological role remains to be elucidated.

Ophthalmologica, 1995, 209(3), 165 - 71
Treatment of postoperative endophthalmitis; Verbraeken H; Despite many advances in microsurgery, asepsis, antibiotics and intraocular lenses, postoperative endophthalmitis continues to be responsible for the loss of many eyes . In a series of 153 cases of endophthalmitis with a positive culture, 115 occurred after ophthalmic surgery . Eyes appear to be more vulnerable to this complication after extracapsular lens extraction in particular . The analysis of the patients operated for cataract in our own department shows that the incidence of endophthalmitis is 3 times higher in the extracapsular group with lens implantation than in the group of intracapsular lens extraction without lens implantation . Quick diagnosis and prompt action are essential to successful treatment . The treatment consists of vitrectomy and has the following three purposes: (1) provision of a good specimen for direct bacteriological examination and culture; (2) removal of toxins and cells, and (3) creation of space for the injection of an antibiotic . The authors found highly divergent prognoses and bacteriological patterns for endophthalmitis following vitrectomy, bleb surgery and cataract . Similar bacteriological agents do not give rise to the same prognosis after different operations . The post-lens implantation group (88 cases) permitted a distinction to be made between intracapsular and extracapsular surgery . A statistically significantly higher percentage of infection by low-virulence organisms (e.g . Staphylococcus epidermidis) was seen after extracapsular surgery: 60% compared to 30% . The analysis shows that this difference in bacteriological spectrum is the sole explanation for the better functional results following extracapsular surgery.

Arch Dermatol Res, 1995, 287(5), 443 - 7
Susceptible responsiveness to bacterial superantigens in peripheral blood mononuclear cells from patients with psoriasis; Yokote R et al.; We investigated the in vitro responses to bacterial superantigens of peripheral blood mononuclear cells taken from patients with psoriasis (one arthropathic, two guttate and four chronic plaque type) . We also analysed the relationship between the magnitude of the responses of peripheral blood mononuclear cells to bacterial exotoxins and the number of circulating T cells bearing V beta 2 and V beta 3 regions . The proliferative response of peripheral blood mononuclear cells to Staphylococcal enterotoxin B and toxic shock syndrome toxin-1 was significantly higher in patients with active psoriasis than in normal subjects . An improvement in skin eruption was associated with a decrease in the lymphocyte response to one-half or one-third that of the active phase . There was no significant difference between patients with psoriasis and normal subjects in the percentage of V beta 2- and V beta 3-positive circulating T cells . The percentages of V beta 2-positive and V beta 3-positive cells were not correlated with the levels of responsiveness to toxic shock syndrome toxin-1 and to Staphylococcal enterotoxin B, respectively . These findings suggest that the magnitude of responses of peripheral blood mononuclear cells to bacterial toxins does not depend on the number of T cells reactive with the relevant superantigen, but depends on the extent of skin lesions in psoriasis.

Nephrol Dial Transplant, 1995, 10(4), 533 - 6
Deep wound infections in simultaneous pancreas-kidney transplant recipients on peritoneal dialysis; Douzdjian V et al.; Peripancreatic abscess formation remains an important cause of surgical morbidity following simultaneous pancreas-kidney (SPK) transplants . The purpose of this study is to define the association, if any, between the pretransplant method of dialysis and the incidence of peripancreatic abscess formation . We reviewed 57 bladder-drained SPK transplants . Of these, 42 were on haemodialysis (HD) and 15 on peritoneal dialysis (PD) prior to transplantation . Two grafts were lost and three patients died as a result of peripancreatic infections for a total graft loss of 8.8% . The incidence of this complication was higher (P = 0.036 Pearson chi square) in patients on PD (40%) prior to transplant when compared to patients on HD (14%) . The most common organism cultured from the abscesses was Staphylococcus epidermidis in both groups . Impairment of peritoneal macrophage function and accumulation of peritoneal effluent as a result of altered peritoneal membrane function are some of the changes that occur following chronic peritoneal dialysis and could explain our findings.

Arch Orthop Trauma Surg, 1995, 114(3), 179 - 82
Gaucher disease--the orthopaedic aspect . Report of seven cases; Tauber C et al.; Seven patients with Gaucher type 1 disease are presented: five female and two male . The orthopaedic problems encountered were: avascular necrosis of both femoral heads in two girls, bilateral bone infarctions of the femurs in all three girls, complicated by staphylococcal osteomyelitis of the right femur in one girl and by pathological fractures of both femurs in another girl, osteonecrosis of both humeral heads in one male adult patient, osteonecrosis of the femoral heads in two further adults (male and female), and a pathological fracture of the fourth lumbar vertebra in another adult woman with concomitant humeral head involvement . The patient with osteonecrosis of the humeral head was treated with a total shoulder arthroplasty, and the two patients with osteonecrosis of the femoral heads were treated with total hip arthroplasties, with satisfactory intermediate and long-term results.

Arch Dermatol Res, 1995, 287(3-4), 254 - 8
Localization of epidermal sphingolipid synthesis and serine palmitoyl transferase activity: alterations imposed by permeability barrier requirements; Holleran WM et al.; Sphingolipids, the predominant lipid species in mammalian stratum corneum play, a central role in permeability barrier homeostatis . Prior studies have shown that the epidermis synthesizes abundant sphingolipids, a process regulated by barrier requirements, and that inhibition of sphingolipid synthesis interferes with barrier homeostasis . To investigate further the relationship between epidermal sphingolipid metabolism and barrier function, we localized sphingolipid synthetic activity in murine epidermis under basal conditions, and following acute (acetone treatment) or chronic (essential fatty acid deficiency, EFAD) barrier perturbation, using dithiothreitol and/or the staphylococcal epidermolytic toxin to isolate the lower from the outer epidermis . Under basal conditions, both the activity of serine palmitoyl transferase (SPT), the rate-limiting enzyme of sphingolipid synthesis, and the rates of 3H-H2O incorporation into sphingolipids were nearly equivalent in the lower and the outer epidermis . Following acute barrier perturbation, SPT activity increased significantly in both the lower (35%; P < 0.05) and the outer epidermal layers (60%; P < 0.01) . The rates of 3H-H2O incorporation into each major sphingolipid family, including ceramides, glucosylceramides and sphingomyelin, increased significantly in both the lower and the outer epidermis of treated flanks after acute barrier disruption . Finally, SPT activity was modestly elevated (20%; P < 0.01) in the lower but not in the outer epidermis of EFAD animals . These studies demonstrate the ability of both lower and outer epidermal cells to generate sphingolipids, and that permeability barrier homeostatic mechanisms appear to differentially regulate SPT activity and sphingolipid synthesis in the lower and the outer epidermis in response to acute and chronic barrier perturbation.(ABSTRACT TRUNCATED AT 250 WORDS)

Biosens Bioelectron, 1995 Fall, 10(8), 675 - 82
Impedance based sensing of the specific binding reaction between Staphylococcus enterotoxin B and its antibody on an ultra-thin platinum film; DeSilva MS et al.; Immunobiosensing techniques to measure specific antigen-antibody binding reactions are important in the development of biosensor applications in biotechnology, in vitro diagnosis, medicine and food technology . An immunobiosensor was constructed to measure the specific binding reaction between Staphylococcus enterotoxin B (SEB) and anti-SEB antibodies . The biosensor comprised an anti-SEB bioactive layer covalently immobilized on an ultra-thin platinum (Pt) film sputtered onto a 100 nm thick silicon dioxide layer on a silicon chip . The Pt film was discontinuous with a normal thickness of 25 A . The impedance of the Pt film decreased during the binding of the anti-SEB to SEB in phosphate buffered saline (PBS) at room temperature . The impedance decreases were irreversible in PBS before saturation of the specific binding sites . When saturated, the impedance at 100 Hz was 14% of the value obtained for the fresh anti-SEB layer in PBS . The magnitude of the impedance (Z) decrease followed a simple relationship with SEB concentration in the range between 0.389 and 10.70 ng/ml SEB . The specificity of the biosensor was demonstrated by showing that no irreversible impedance decreases occurred when the sensor was exposed to 100 ng/ml kappa-casein, or alpha-lactalbumin, in PBS.

Microbiol Immunol, 1995, 39(5), 315 - 9
Typing of Staphylococcus epidermidis colonizing in human nares by pulsed-field gel electrophoresis; Hu L et al.; From the nares of 11 healthy adults, 253 strains of coagulase negative staphylococcus were isolated and 88% of them were identified as Staphylococcus epidermidis using the API STAPH system . Chromosomal DNA fingerprinting of the isolated strains revealed that each person carried multiple types of S . epidermidis in his or her nares . The colonization of the strains was not stable; the types of the isolates changed in the first and the second examinations 5 months apart . The results contrasted with previous findings in which only one strain of S . aureus colonized persistently in the nares of healthy adults.

Drugs Exp Clin Res, 1995, 21(2), 65 - 70
Evaluation of in vitro and in vivo antimicrobial activity of a new topical antiinfective agent; D'Urso CM et al.; ST 1103 (Undecyl {4-N,N,N-trimethylammonium-(R)-3- isovaleroyloxy}-butanoate methanesulfonate) is a novel compound endowed with a broad antimicrobial spectrum . ST 1103 is able to inhibit the in vitro growth of Gram-positive bacteria (mean MIC value of 2.60 micrograms/ml), Gram-negative bacteria (mean MIC value of 27.00 micrograms/ml), yeasts and yeast-like fungi (mean MIC value of 3.76 micrograms/ml), filamentous and dermatophytic fungi (mean MIC value of 18.33 micrograms/ml) . Since indirect evidence indicates a poor oral absorbtion, ST 1103 was topically administered to mice with skin infections caused by mixed inocula . In these conditions, ST 1103 was able to cure mice infected with T . quinckeanum, S . aureus as well as immunodepressed mice infected with T . quinckeanum, S . aureus and C . albicans . Conversely, miconazole (reference compound) appeared inadequate, in our experimental conditions, for a definitive therapy of the skin mycosis superinfected by staphylococcus . By using an in vitro 3D-human skin model, ST 1103 was fairly well tolerated in terms of both cell viability and release of inflammatory mediators . In a dermal tolerance study in mice, ST 1103 at a concentration of 1% did not show any sign of local irritation on both intact and abraded skin after an 8-day topical treatment . In conclusion, ST 1103 appears to be a promising candidate for treatment of cutaneous infections caused by mixed microbial pathogens.

J Exp Med, 1995 Jan 1, 181(1), 71 - 7
Fas ligand mediates activation-induced cell death in human T lymphocytes; Alderson MR et al.; A significant proportion of previously activated human T cells undergo apoptosis when triggered through the CD3/T cell receptor complex, a process termed activation-induced cell death (AICD) . Ligation of Fas on activated T cells by either Fas antibodies or recombinant human Fas-ligand (Fas-L) also results in cytolysis . We demonstrate that these two pathways of apoptosis are causally related . Stimulation of previously activated T cells resulted in the expression of Fas-L mRNA and lysis of Fas-positive target cells . Fas-L antagonists inhibited AICD of T cell clones and staphylococcus enterotoxin B (SEB)-specific T cell lines . The data indicate AICD in previously stimulated T cells is mediated by Fas/Fas-L interactions.

Acta Vet Scand, 1995, 36(3), 335 - 42
Antimicrobial susceptibility and rRNA gene restriction patterns among Staphylococcus intermedius from healthy dogs and from dogs suffering from pyoderma or otitis externa; Pedersen K et al.; A total of 60 Staphylococcus intermedius strains from dogs were investigated by their sensitivity to various antibiotics (50 strains) and by their rRNA gene restriction patterns (ribotyping) (60 strains) . Fifteen isolates were from healthy dogs, 9 with otitis externa, and 36 with pyoderma, including 10 strains from a previous study . Sixty per cent of the 50 strains tested for antibiotic susceptibility demonstrated resistance to penicillin, 24% to spiramycin, 20% to tetracycline, 16% to chloramphenicol, and 2% to fucidic acid . All isolates were susceptible to amoxycillin with clavulanic acid, enrofloxacin, and sulphonamides with trimethoprim . There were no significant differences in antimicrobial susceptibility patterns observed among isolates from pyoderma, otitis externa or healthy dogs . Among the 60 strains studied by ribotyping, 10 different ribotypes were identified: 6 different ribotypes among isolates from otitis externa, 8 among isolates from pyoderma, and 5 among isolates from healthy dogs . One ribotype (profile C) was dominant among the isolates from healthy dogs while another ribotype (profile A) was dominant among strains from dogs suffering from pyoderma . This profile was not demonstrated in any of the strains from healthy dogs . From 5 different dogs suffering from pyoderma, 2 different clones were demonstrated based on their plasmid profile and antibiogram . In these dogs 1 of the clones always belonged to ribotype A . The results concerning strains of S . intermedius isolated from furunculosis suggest the existence of distinct subpopulations with different pathogenicity to dogs.

Pediatr Nephrol, 1995, 9 Suppl, S12 - 7
Infectious and catheter-related complications in pediatric patients treated with peritoneal dialysis at a single institution; Kuizon B et al.; Continuous ambulatory peritoneal dialysis (CAPD) and continuous cycling peritoneal dialysis (CCPD) are the predominant dialytic modalities for the majority of children while awaiting transplantation . Wide acceptability of peritoneal dialysis is hindered by infectious complications . A retrospective review of 367 pediatric patients treated with CAPD/CCPD for at least 3 months from September 1980 through December 1994 revealed that the peritonitis incidence ranged from 1.7 to 0.78 episodes per patient-year . No differences in peritonitis rates were observed between patients treated with CAPD or CCPD . Gram-positive organisms were responsible for the majority of peritonitis episodes . Age, sex, race, primary renal disease, presence of nephrotic syndrome, and serum albumin level were not associated risk factors . Longer time on treatment and diminished serum IgG level were associated with increased peritonitis incidence . Treatment was successfully completed at home in most cases . Almost half of the catheter losses were caused by Staphylococcus, Pseudomonas, and fungal peritonitis and tunnel/exit-site infections . Infectious complications are still the major causes of morbidity and treatment failure in patients treated with CAPD/CCPD . Thus, controlled studies are needed to assess methods for prevention or improvement of peritonitis rates in this patient population.

Parasitol Res, 1995, 81(7), 560 - 4
Circulating trans-sialidase activity and trans-sialidase-inhibiting antibodies in Trypanosoma cruzi-infected mice; Alcantara-Neves NM et al.; Parasite-derived trans-sialidase (TS) activity was demonstrated in the serum and blood of Trypanosoma cruzi-infected mice . Serum TS activity levels correlated well with parasitemia in BALB/c and Swiss mice during the initial stages of the infection . However, in later stages the TS activity levels decreased despite increasing parasitemia . This coincided with the appearance of circulating TS antibodies . On the other hand, there was always a good correlation between TS activity and parasitemia in athymic nude mice . Sera from mice with high parasitemia and low TS activity inhibited TS activity in vitro . The inhibition was also observed with purified serum IgG, and it was absorbed by staphylococcal protein A, indicating that it was caused by anti-TS IgG antibodies . These antibodies inhibited the enzymatic activity of insolubilized TS, indicating that they act by interfering with the catalytic site rather than by aggregating the enzyme . The presence of inhibitory antibodies, however, did not prevent the progression of parasitemia in BALB/c mice.

Gene, 1994 Dec 30, 151(1-2), 45 - 51
Construction and characterization of M13 bacteriophages displaying functional IgG-binding domains of staphylococcal protein A; Kushwaha A et al.; Staphylococcal protein A (SPA) is ranked as a versatile probe in immunoassays because of its immunoglobulin G (IgG)-binding capability . However, poor binding of SPA to the IgG of some laboratory animals and its inability to bind human IgG3 restricts its universal utility . In the present study, DNA encoding the four IgG-binding domains of SPA (E, D, A and B) or the B domain alone has been fused, in separate phagemid vectors, to the 5' end of gene 111 of the phage M13 . Upon infection by helper phage M13KO7, phagemid particles encapsulating single-stranded DNA were produced . Dot immunoblot and Western blot analyses showed the presence of fusion proteins on the M13 surface . Binding of rabbit IgG-horseradish peroxidase (IgG-HRP) complex to the phage particles confirmed that the fusion proteins possessed functional IgG-binding domains . The interaction of these phages with immobilised human IgG and its various subclasses was studied by the phage capture immunoassay where the captured phages were detected by a monoclonal antibody to the major coat protein encoded by gene VIII (gVIII) . The phages showed maximal binding to IgG1 kappa, followed by IgG2 kappa, and showed negligible binding to the IgG3 kapa and IgG3 lambda subclasses . The specificity of IgG-binding phages was confirmed in a phage capture and elution assay where the binding of these phages to immobilised human IgG1 kapa weas abolished in the presence of excess of soluble protein A . Moreover, IgG-binding phages could be enriched approx . 1000-fold over non-specific phages in a single round of panning.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1994 Dec 23, 269(51), 32063 - 9
Induction of chemokine gene expression by major histocompatibility complex class II ligands in human fibroblast-like synoviocytes . Differential regulation by interleukin-4 and dexamethasone; Mehindate K et al.; Chemokines play a key role in recruiting leukocytes into inflamed synovial environment, and the cells of the synovial membrane, which express high levels of major histocompatibility complex (MHC) class II molecules, are a major source of these chemokines . Our data indicated that engagement of MHC class II molecules by staphylococcal enterotoxin A superantigen resulted in the induction of chemokine gene expression as well as protein synthesis . Pretreatment of the cells with cycloheximide potentiated the effect of superantigen on chemokine mRNA induction, suggesting that the expression of these genes may occur independently of prior protein synthesis . Ligation of MHC class II molecules in fibroblast-like synoviocytes by other ligands such as Mycoplasma arthritidis-derived superantigen and anti-class II antibody could also trigger an increase in the mRNA level of RANTES, MCP-1, and interleukin (IL)-8 . The addition of dexamethasone to superantigen-treated fibroblast-like synoviocytes inhibited the mRNA expression of all three chemokines . IL-4 treatment decreased only the stimulating effect of superantigen on RANTES messanger suggesting that different mechanisms are involved in regulating these genes . The inhibitory effect of dexamethasone did not require a de novo protein synthesis, whereas that of IL-4 was protein-dependent . This report demonstrates that MHC class II ligands (superantigens and anti-MHC class II antibodies) may represent an important agent by which inflammatory chemokines can be induced and shows that this response can be modulated by the anti-inflammatory agents dexamethasone and IL-4.

Proc Natl Acad Sci U S A, 1994 Dec 20, 91(26), 12828 - 31
Subunit stoichiometry of staphylococcal alpha-hemolysin in crystals and on membranes: a heptameric transmembrane pore; Gouaux JE et al.; Elucidation of the accurate subunit stoichiometry of oligomeric membrane proteins is fraught with complexities . The interpretations of chemical cross-linking, analytical ultracentrifugation, gel filtration, and low-resolution electron microscopy studies are often ambiguous . Staphylococcal alpha-hemolysin (alpha HL), a homooligomeric toxin that forms channels in cell membranes, was believed to possess six subunits arranged around a sixfold axis of symmetry . Here, we report that analysis of x-ray diffraction data and chemical modification experiments indicate that the alpha HL oligomer is a heptamer . Self-rotation functions calculated using x-ray diffraction data from single crystals of alpha HL oligomers show a sevenfold axis of rotational symmetry . The alpha HL pore formed on rabbit erythrocyte membranes was determined to be a heptamer by electrophoretic separation of alpha HL heteromers formed from subunits with the charge of wild-type alpha HL and subunits with additional negative charge generated by targeted chemical modification of a single-cysteine mutant . These data establish the heptameric oligomerization state of the alpha HL transmembrane pore both in three-dimensional crystals and on a biological membrane.

Proc Natl Acad Sci U S A, 1994 Dec 20, 91(26), 12654 - 8
Functional three-domain single-chain T-cell receptors; Chung S et al.; T-cell receptors (TCRs) are membrane anchored heterodimers structurally related to antibody molecules . Single-chain antibodies can be engineered by linking the two variable domains, which fold properly by themselves . However, proper assembly of the variable domains of a human TCR (V alpha and V beta) that recognize the HLA-DR2b/myelin basic protein-(85-99) peptide complex was critically dependent on the addition of a third domain, the constant region of the TCR beta chain (C beta), to the single-chain construct . Single-chain molecules with the three-domain design, but not those with the two-domain design, expressed in a eukaryotic cell as chimeric molecules linked either to glycosyl phosphatidylinositol or to the transmembrane/cytoplasmic domains of the CD3 zeta chain were recognized by a conformation-sensitive monoclonal antibody . The chimeric three-domain single-chain TCR linked to CD3 zeta chain signaled in response to both the specific HLA-DR/peptide and the HLA-DR/superantigen staphylococcal enterotoxin B complexes . Thus, by using this three-domain design, functional single-chain TCR molecules were expressed with high efficiency . The lipid-linked single-chain TCR was solubilized by enzymatic cleavage and purified by affinity chromatography . The apparent requirement of the constant domain for cooperative folding of the two TCR variable domains may reflect significant structural differences between TCR and antibody molecules.

Blood, 1994 Dec 15, 84(12), 4234 - 41
A defect in the early phase of T-cell receptor-mediated T-cell activation in patients with common variable immunodeficiency; Fischer MB et al.; Common variable immunodeficiency (CVID) is characterized by an impairment of specific antibody production and a decrease in all or selected Ig isotypes . Abnormalities at the level of the B cells, T cells, and antigen-presenting cells have been described . In the present study, we have focused our attention on T-cell activation in CVID . T cells from 15 of 24 patients failed to respond to recall antigens (eg, tetanus toxoid, Escherichia coli) . Of these 15 patients, 11 were studied in detail and showed significantly decreased T-cell proliferative responses and/or decreased interleukin-2 and interferon-gamma production on T-cell receptor-mediated stimulation with recall antigens and superantigens (staphylococcal enterotoxins {SE}); however, T-cell response to mitogens (anti-CD3 monoclonal antibody, phytohemagglutinin) was normal . The defect in interleukin-2 and interferon-gamma release on tetanus toxoid stimulation could also be documented in purified CD4 T cells of the patients and was present in patients with high and normal CD8 counts alike . Furthermore, patients' T cells failed to mount a significant elevation in free intracellular calcium (Ca++ flux) in response to superantigen, whereas the response to phorbol myristate acetate and ionomycin, bypassing receptor-mediated signaling, was unimpaired . These results indicate a defect in the early phase of T-cell activation after triggering of the T-cell receptor in a significant subgroup of CVID patients.

Eur J Biochem, 1994 Dec 15, 226(3), 847 - 52
Effects of amino acid insertions on the proteolysis of a staphylococcal protein A derivative in Escherichia coli; Yang S et al.; In vivo proteolysis of protein ZZT0, derived from the B domain of staphylococcal protein A, was investigated in Escherichia coli before and after insertion of 1-3 multiples of the tetrapeptide Ala-Trp-Trp-Pro close to the C-terminus of ZZT0 . Before insertion, ZZT0 was proteolytically stable as judged from the purity of IgG binding proteins up to 1 h after inhibition of protein synthesis with chloramphenicol . Insertion of 1-3 units of Ala-Trp-Trp-Pro into ZZT0 increased progressively the sensitivity to proteolysis and induced DnaK and GroEL binding to the protein . The time for 50% in vivo hydrolysis of the full length protein derivative that was most susceptible to proteolysis, i.e . with three tetrapeptide units, was about 40 min when cultivated in a bioreactor and about 4 min in a shaken flask culture . Molecular masses and N-terminal sequences of the main degradation products indicated that protein ZZT0 is cleaved at identical sites irrespective of the number of inserted tetrapeptide units and that the cleavage sites are located far from the insertion point . Insertion of another hydrophobic amino acid, isoleucine, as the tetrapeptide Ala-Ile-Ile-Pro, only induced a slight proteolysis of the ZZT0 molecule under similar conditions . This indicates that the insertion of tryptophan residues, rather than of a general hydrophobic segment, plays an essential role in the induced proteolysis of the ZZT0 protein.

FEBS Lett, 1994 Dec 12, 356(1), 66 - 71
Surface labeling of key residues during assembly of the transmembrane pore formed by staphylococcal alpha-hemolysin; Krishnasastry M et al.; Structural changes in staphylococcal alpha-hemolysin (alpha HL) that occur during oligomerization and pore formation on membranes have been examined by using a simple gel-shift assay to determine the rate of modification of key single-cysteine mutants with the hydrophilic sulfhydryl reagent, 4-acetamido-4'-((iodoacetyl)amino)stilbene-2,2'-disulfonate (IASD) . The central glycine-rich loop of alpha HL lines the lumen of the transmembrane channel . A residue in the loop remains accessible to IASD after assembly, in keeping with the ability of the pore to pass molecules of approximately 1000 Da . By contrast, residues near the N-terminus, which are critical for pore function, become deeply buried during oligomerization, while a residue at the extreme C-terminus increases in reactivity after assembly, consistent with a location in the part of the pore that projects from the surface of the lipid bilayer.

Eur J Pharmacol, 1994 Dec 12, 271(1), 131 - 9
Evidence for the formation of endothelin by lysed red blood cells from endogenous precursor; Tippler B et al.; The release of endothelin from various blood cell fractions was investigated . Human as well as rat blood cell fractions homogenized by sonification were incubated in buffer for up to 60 min . Neither in platelet nor leukocyte homogenates from either species could immunoreactive endothelin be detected . In contrast, homogenates of red blood cells from both species showed a rapid and time-dependent rise of immunoreactive endothelin levels, reaching a peak at 15 min and decreasing thereafter . However, at time point 0 no immunoreactive endothelin could be detected . Reverse phase high performance liquid chromatography showed immunoreactive endothelin to consist of endothelin-1 as well as big endothelin-1 . The release of immunoreactive endothelin in human and rat homogenates was concentration-dependently inhibited by the protease inhibitors, leupeptin, phosphoramidon, chymostatin and pepstatin A in order of increasing potency . Intact red blood cells did not incorporate {125I}endothelin-1 nor did they transform exogenous big endothelin-1 to endothelin-1 . However, haemolysis of red blood cells with hypotonic saline (0.2%) or incubation with pore-forming staphylococcal alpha-toxin induced the release of immunoreactive endothelin into the buffer samples . Thus, apart from the indirect vasoconstrictor, haemoglobin, red blood cells can also liberate the direct vasoconstrictor, endothelin, a finding expected to be of considerable pathophysiological significance.

J Biol Chem, 1994 Dec 9, 269(49), 30994 - 8
The nucleic acid binding activity of nucleolar protein B23.1 resides in its carboxyl-terminal end; Wang D et al.; Protein B23 is a major nucleolar phosphoprotein proposed to be a ribosome assembly factor . Protein B23 exists as two isoforms, B23.1 and B23.2, differing only in their carboxyl-terminal sequences . The interaction of recombinantly produced B23 isoforms with double-stranded DNA was studied using gel retardation and nitrocellulose filter disk assays . Protein B23.1 bound saturably to radiolabeled plasmid DNA . By competition assays protein B23.1 was also capable of binding RNA and single-stranded DNA . On the other hand, protein B23.2, the shorter of the two isoforms, was not capable of binding double-stranded DNA . The latter result suggested that the carboxyl-terminal end of B23.1 is essential for DNA binding activity . This was confirmed by partial digestion experiments using staphylococcal V8 protease which showed that a 5-kDa fragment, containing the carboxyl-terminal end of protein B23.1 retained DNA binding activity similar to that of the parent molecule . In contrast, a 19-kDa fragment from the amino-terminal half of B23.1 did not bind DNA . The sequence of the carboxyl-terminal 68 amino acids comprising the 5-kDa fragment showed little, if any, similarity to other proteins, suggesting that this segment contains a previously undiscovered nucleic acid binding motif.

J Comp Neurol, 1994 Dec 8, 350(2), 260 - 71
Immunocytochemical localization of the beta 2 subunit of the gamma-aminobutyric acidA receptor in the rat brain; Moreno JI et al.; An antiserum to the beta 2 subunit of the rat gamma-aminobutyric acid (GABAA) receptor was prepared by immunizing a rabbit with a fusion protein expressed in bacteria . The fusion protein had the large, intracellular loop expanding between the putative M3 and M4 transmembrane domains of the beta 2 subunit fused to staphylococcal protein A (SPA) . The antiserum immunoprecipitated both the solubilized and the affinity-purified GABAA receptors . The anti-beta 2 antibodies were affinity purified on immobilized beta 2 intracellular loop peptide . The antibodies recognized a 55-57 kDa peptide in immunoblots of either crude membranes from rat cerebral cortex or affinity-purified GABAA receptors from bovine cerebral cortex . Immunocytochemistry with the affinity-purified antibody has revealed for the first time the localization of the beta 2 subunit in the rat brain . A comparative study of the regional and cellular immunoreactivities of the affinity-purified anti-beta 2 antibody and the monoclonal antibody 62-3G1 (which recognizes both beta 2 and beta 3 subunits) is presented . The procedure described for generating and preparing specific anti-beta 2 subunit antibodies that are valuable for immunocytochemistry could be extended to other GABAA receptor subunits.

J Immunol Methods, 1994 Dec 2, 176(2), 213 - 20
The generation of monoclonal antibodies recognising novel epitopes by immunisation with solid matrix antigen-antibody complexes reveals a polymorphic determinant on feline CD4; Willett BJ et al.; Monoclonal antibodies were generated against the feline homologue of CD4 (fCD4) by immunisation of mice with solid matrix antigen-antibody complexes of monoclonal antibody Fel7 (anti-fCD4) and formalin-fixed Staphylococcus A (SMAA-fCD4) . The resulting fusion produced nine monoclonal antibodies each of which recognised a major population of feline lymphocytes and which immunoprecipitated a 55 kDa ligand from the feline T lymphosarcoma cell line 3201 . Epitope mapping of the antibodies against soluble fCD4 by surface plasmon resonance indicated that the antibodies recognised five separate epitopes distinct from that defined by the Fel7 antibody used to prepare the SMAA-fCD4 . These data demonstrate that SMAA complexes are an efficient means of generating monoclonal antibodies recognising novel epitopes on an antigen . One monoclonal antibody (vpg39) recognised an epitope that was expressed variably between cats, being either present or completely absent . Analysis of peripheral blood lymphocytes from specific pathogen free cats suggested that failure to react with the vpg39 antibody was an inherited trait.

Cell Immunol, 1994 Dec, 159(2), 315 - 22
Noninvolvement of V beta 8+ T cells in murine thyroglobulin-induced experimental autoimmune thyroiditis; Fuller BE et al.; In experimental autoimmune thyroiditis (EAT) induced with mouse thyroglobulin (MTg), T cell receptor (TCR) V beta gene usage in the pathogenesis of disease is unknown . We report here studies evaluating V beta 8 gene usage in EAT, as V beta 8+ T cells are reportedly involved in some experimental autoimmune diseases . Spleen cells (SC) from MTg-immunized CBA/J (H-2k) mice were activated in vitro for adoptive transfer into syngeneic recipients . Elimination of V beta 8+ T cells by treating recipients with V beta 8 monoclonal antibody (mAb) following transfer of MTg-activated SC did not reduce disease severity . Conversely . MTg-primed SC were stimulated in vitro with V beta 8 mAb or staphylococcal enterotoxin B, which activates V beta 8+ T cells in CBA/J mice . Neither activated population transferred disease, in contrast to cells activated with MTg . Thus, in MTg-induced EAT, V beta 8+ T cells do not play a major role in pathogenesis.

Arch Surg, 1994 Dec, 129(12), 1310 - 6; discussion 1316-7
Characterization and impact of wound infection after pancreas transplantation; Everett JE et al.; OBJECTIVE: To characterize the incidence, microbial pathogenesis, risk factors, and impact of wound infection after pancreas transplantation . DESIGN: Retrospective analysis . SETTING: A large university hospital . PATIENTS: From January 1, 1990, to September 30, 1993, 197 patients underwent 207 consecutive pancreas transplantation procedures . MAIN OUTCOME MEASURES: Wound infection and patient and allograft survival rates at 1 year . RESULTS: Sixty-nine patients (33%) suffered wound infections: 21 (10%) were superficial; 31 (15%), deep; and 17 (8%), combined . Most (74%) wound infections were monomicrobial . Staphylococcus epidermidis and Candida species were the most common pathogens . Prolonged operating time, older donors, and enteric drainage were associated with higher wound infection rates . Deep and combined wound infections led to allograft loss despite subsequent salvage procedures . Combined wound infection was associated with significantly higher mortality . CONCLUSIONS: A deep wound infection should be an indication for allograft removal . Antifungal prophylaxis, stringent donor criteria, and delayed primary wound closure should lower the incidence of wound infection.

J Immunol, 1994 Dec 1, 153(11), 4853 - 61
Activation of T cells by superantigen in class II-negative mice; Avery AC et al.; The ability of the staphylococcal enterotoxins to stimulate T cells has been thought to depend on their association with class II MHC products . Here, we demonstrate that a subgroup of staphylococcal enterotoxins, which includes staphylococcal enterotoxin C and staphylococcal enterotoxin E, stimulates strong MHC-independent responses, thereby resulting in T cell expansion and generation of CTL . The immunologic consequences of MHC-independent activation of T cells by superantigens differ from those of class II-dependent activation, inasmuch as this pathway does not result in detectable T cell deletion . These findings delineate a novel MHC-independent T cell activation pathway that leads to both clonal expansion and expression of CTL effector function in response to a subgroup of bacterial superantigens.

Pediatr Dermatol, 1994 Dec, 11(4), 331 - 4
Toxic epidermal necrolysis associated with Klebsiella pneumoniae sepsis; Picard E et al.; Toxic epidermal necrolysis (TEN) is a rare condition in childhood usually attributed to drugs . We describe a 4-month-old infant who developed typical clinical and histologic findings of TEN concomitantly with Klebsiella pneumoniae sepsis . We emphasize that in cases of acute, severe exfoliative disease in infants, apart from staphylococcal infection, gram-negative bacterial sepsis must also be considered.

Clin Infect Dis, 1994 Dec, 19(6), 1138 - 40
Staphylococcus epidermidis meningitis and an intraspinal abscess associated with a midthoracic dermal sinus tract; Gurbani SG et al.; Congenital neuroectodermal defects are associated with meningitis that is unresponsive to conventional antibiotic therapy, recurrent bacterial meningitis, or meningitis due to an organism that is not usually the cause of this disease . Midthoracic dermoid and epidermoid cysts occur rarely and are easily overlooked . We report the case of a 13-month-old boy with meningitis that was unresponsive to antimicrobial therapy . We subsequently identified an intraspinal abscess, an infection due to Staphylococcus epidermidis, and a midthoracic dermoid cyst . We emphasize the need for clinicians to have a high index of suspicion and to make a careful physical examination when antibiotic treatment fails in patients with meningitis . We also provide a detailed anatomy of a midthoracic dermoid cyst and illustrate the usefulness of magnetic resonance imaging in preoperative diagnosis of congenital neuroectodermal defects.

Am J Vet Res, 1994 Dec, 55(12), 1690 - 6
Immunomodulatory effects of staphylococcal antigen and antigen-antibody complexes on canine mononuclear and polymorphonuclear leukocytes; DeBoer DJ; Staphylococcal antigens and immune complexes (IC) prepared from antigen and hyperimmune canine serum were tested for their effects on certain functions of mononuclear (MN) and polymorphonuclear (PMN) leukocytes (cells) obtained from healthy dogs . The effect on MN cells was studied by determining the ability of antigen or IC to augment or inhibit mitogenesis induced by phytohemagglutinin (PHA) . The effect of antigen or IC on PMN cells was studied by measurement of H2O2 production as an indicator of respiratory burst . Neither the antigen nor the IC, when cultured with MN cells, was mitogenic . Coincubation of antigen or IC with MN cells and PHA resulted in a concentration-dependent decrease in mitogenesis . The decreased mitogenesis could not be overcome by addition of excess PHA, and may in part have been related to toxic effects of the antigen or IC on MN cells . When MN cells were instead preincubated with antigen or IC, then washed and stimulated with PHA, there was still a concentration-dependent inhibition of mitogenesis, although toxicity to the cells was not observed . Low concentrations of staphylococcal antigen or IC stimulated slight H2O2 production by PMN cells . When PMN cells were coincubated with IC and another stimulus (opsonized zymosan or phorbol myristate acetate), IC appeared to augment phorbol myristate acetate-, but not zymosan-induced stimulation . These results suggest that staphylococcal antigens, either alone or complexed with antibody, have the ability to stimulate PMN cells and inhibit MN cell function . Such actions may have a role in the pathogenesis of recurrent staphylococcal infection in canine patients.

Burns, 1994 Dec, 20(6), 499 - 502
Staphylococcal toxic shock syndrome in a paediatric burn unit; Bacha EA et al.; A case of staphylococcal toxic shock syndrome (TSS) in a 15-year-old patient undergoing reconstructive burn surgery is reported . This syndrome, first described in menstruating women using tampons, can also occur as a postsurgical complication . The epidemiology, microbiology, clinical presentation and management of non-menstrual TSS are discussed and correlated with the patient present . We advise caution in the use of occlusive dressings and recommend constant isolation of patients with open wounds colonized with a TSST-1 producing strain of Staph . aureus.

Immunology, 1994 Dec, 83(4), 568 - 72
Langerhans' cell depletion by staphylococcal superantigens; Pickard S et al.; Superantigens were examined for effects on the distribution of Langerhans' cells (LC) in mouse skin . This was accomplished by analysing the expression of LC-specific markers, ATPase and IA among the epidermal portion of cultured sections of mouse skin following treatment with staphylococcal enterotoxins . In this study, treatment of skin sections with staphylococcal enterotoxin A or exfoliative toxin but not toxic shock syndrome toxin led to significant depletion of LC . This depletion was inhibited by agents which specifically block the action of GTP binding proteins or their associated kinases (cholera and pertussis toxins and H-8) as well as those which block protein or RNA synthesis . Therefore, signals which lead to LC depletion in response to staphylococcal enterotoxins appear to involve a cholera and pertussis toxin-sensitive GTP-binding protein and protein synthesis . These requirements are identical to those observed previously for LC depletion following exposure of skin to ultraviolet radiation.

Enferm Infecc Microbiol Clin, 1994 Dec, 12(10), 490 - 6
{Hip prosthesis infection: diagnostic approach and treatment of 27 cases}; Salavert M et al.; BACKGROUND: Preoperative diagnosis of hip prosthesis infection (HPI) is difficult . There is no therapeutic option which is completely effective and without risk . The aim of this study was to evaluate a diagnostic approach and therapeutic strategy in a group of patients with HPI . PATIENTS AND METHODS: A retrospective study of 27 episodes of HPI diagnosed by anatomopathologic and/or microbiologic examination of surgical samples was performed . RESULTS: Twenty-three patients with 27 episodes of HPI out of a total of 24 hip prosthesis (HP) were treated . The infection was early in 15 episodes . The etiologic agents were plasmocoagulase negative staphylococcus (NSP) in 11 cases, P . aeruginosa in 8, S . aureus in 5, Enterococcus sp . in 2 and miscellaneous in the remaining cases . In 2 cases the infection was polymycrobial . Following a mean follow period of 22.6 +/- 15.2 months, 13 out of the 14 patients in whom the prosthesis was withdrawn were cured (in 4 a second prosthesis was implanted), one out of 6 in those in whom the prosthesis remained in situ following debridement, and 2 out of 3 episodes in whom reimplantation was performed over time . The withdrawal of the prosthesis was significantly greater than debridement in the treatment of early infection (p < 0.001) . The total mean length of postoperative antibiotherapy was 48.2 +/- 17 days . No differences were observed in the oral versus parenteral treatment (p = 0.22), and nor was prognosis worse in those treated for less than 42 days . CONCLUSIONS: The authors' experience suggests that attempts to save a hip prosthesis in early infection usually fail . In addition to prosthesis withdrawal or implantation of another prosthesis, six weeks of postoperative antibiotic therapy, which may be oral route, appear to be sufficient.

QJM, 1994 Dec, 87(12), 755 - 7
Staphylococcal tricuspid valve endocarditis in patients with structurally normal hearts and no evidence of narcotic abuse; Clifford CP et al.; We report four cases of staphylococcal tricuspid valve endocarditis in patients with structurally normal hearts and no evidence of intravenous drug abuse . The only risk factor was superficial skin sepsis in three of these patients . Medical therapy was successful in all four cases.

Arch Pediatr, 1994 Dec, 1(12), 1106 - 10
{Necrotizing tracheobronchitis in ventilated newborn infants}; Francoise M et al.; BACKGROUND--Necrotizing tracheobronchitis is a severe complication observed in some mechanically ventilated neonates . CASE REPORT--A twin premature (GA = 31 weeks), weighing 1,500 g required oral endotracheal intubation for mechanical ventilation because he suffered from respiratory distress syndrome . He was given indomethacin on day 4 for patent ductus arteriosus . Progressive weaning of ventilation on day 9 was dramatically complicated by hypoxia, respiratory acidosis and right pneumothorax . Immediate endoscopy showed total obstruction of trachea by necrotic secretions the suction of which was followed by rapid improvement of the respiratory condition . Bacterial examination of secretions showed coagulase-negative staphylococcus . The patient was given steroids + antibiotics . Prolonged ventilation resulted in bronchopulmonary dysplasia and the patient was only extubated at week 12 after a normal endoscopic control . CASE REPORT--A premature girl (GA = 32 weeks), weighing 1,800 g required oral endotracheal intubation for mechanical ventilation because she suffered from respiratory distress syndrome . The respiratory condition worsened on day 3, requiring tracheography which showed distal tracheal obstruction . Immediate endoscopy showed thin, adherent and necrotic membranes which were removed by suction . The patient was given steroids +antibiotics and was extubated on day 14 after a normal endoscopic control . CONCLUSIONS--This iatrogeneous complication must be recognized in a ventilated infant when the respiratory condition dramatically worsens . Emergency bronchoscopy permits endotracheal suction of necrotic secretions.

J Antibiot (Tokyo), 1994 Dec, 47(12), 1417 - 24
Bioxalomycins, new antibiotics produced by the marine Streptomyces sp . LL-31F508: taxonomy and fermentation; Bernan VS et al.; An actinomycete strain designated LL-31F508 was isolated from an intertidal sediment sample collected in Key West, Florida . Culture LL-31F508 was assigned to the Streptomyces genus based on the presence of LL-diaminopimelic acid (DAP) in the cell wall and observations of spiny spores using scanning electron microscopy (SEM) . Excellent antimicrobial activity against Staphylococcus and Enterococcus spp . were detected in both the supernatant and cell extract samples from fermentations of culture LL-31F508 . Production of antibiotic activity peaked at 48-50 hours and closely paralleled cell growth, during which time glucose was more rapidly assimilated than dextrin . A series of new antibiotics called the bioxalomycins was identified as the antibacterial products from fermentations of this culture . Fermentation conditions for production of bioxalomycin alpha differed substantially from those required for production of a related compound, naphthyridinomycin, by the reference culture Streptomyces lusitanus NRRL 8034.

Biophys Chem, 1994 Dec, 53(1-2), 15 - 25
The production and X-ray structure determination of perdeuterated Staphylococcal nuclease; Gamble TR et al.; Staphylococcal Nuclease (SNase) has been chosen as a model protein system to evaluate the improvement in neutron diffraction data quality using fully perdeuterated protein . Large quantities of the protein were expressed in Escherichia coli grown in medium containing deuterated amino acids and deuterated water (D2O) and then purified . The mean perdeuteration level of the non-exchangable sites in the protein was found to be 96% by electrospray ionization mass spectrometry . The perdeuterated enzyme was crystallized and its X-ray structure determined . Crystals of perdeuterated SNase have been grown to 1.5 mm3 . Crystallization conditions, space group and cell parameters were found to be the same for both native and perdeuterated forms of the protein . Comparison of these two forms of SNase revealed no significant structural differences between them at the atomic resolution of 1.9 A . Data collection using crystals of the perdeuterated protein is scheduled at the Brookhaven High Flux Beam Reactor.

Appl Environ Microbiol, 1994 Dec, 60(12), 4332 - 8
Construction of an expression system for engineering of the lantibiotic Pep5; Bierbaum G et al.; Pep5 is a lanthionine-containing antimicrobial peptide which is produced by Staphylococcus epidermidis 5 . Its structural gene, pepA, is located on the 20-kb plasmid pED503 . A 6.2-kb fragment of pED503 containing pepA, the immunity gene pepI, and 5.4 kb of downstream sequence was able to direct biosynthesis of biologically active Pep5 in a nonproducing variant of the producer strain which is devoid of pED503 . In addition to producing wild-type Pep5 with a molecular mass of 3,488 Da, the clone produced a peptide with an eightfold-lower bactericidal activity and a mass of 3,506 Da, indicative of incomplete dehydration of one hydroxyamino acid . For construction of the expression system, this 6.2-kb fragment was cut into a 1.39-kb fragment containing pepA and pepI and a 4.8-kb fragment covering the remaining downstream region . This 4.8-kb fragment was directly cloned into an Escherichia coli-Staphylococcus shuttle vector, yielding a new plasmid (pGB9) into which mutated pepA genes generated on the 1.39-kb fragment can be reinserted to yield a functional Pep5 biosynthesis gene cluster . To test the expression system, two mutants were constructed . Lys-18-Pro Pep5 was produced in its dehydrated form and a partially hydrated form in amounts comparable to those of the wild-type peptide . In contrast, only small amounts of Phe-23-Asp Pep5 were excreted, indicating that some residues in the propeptide part of the prelantibiotic may be crucial for certain steps in the biosynthetic pathway of lantibiotics.

Eur J Immunol, 1994 Dec, 24(12), 3140 - 7
Inhibition of the development of immediate hypersensitivity by staphylococcal enterotoxin B; Saloga J et al.; We investigated the ability of staphylococcal enterotoxin B (SEB) to modify the immediate hypersensitivity response induced in BALB/c mice following sensitization to ovalbumin (OVA), a response mediated by OVA-reactive V beta 8 T cells . Mice were sensitized by skin painting with OVA every second day over a period of 2 weeks . SEB, a potent activator of V beta 8+ T cells, was administered at the same site where OVA was applied (skin of the lower abdomen) following two different protocols . In protocol (A) SEB was injected intradermally 1 day before painting with OVA and on day 7; in protocol B, SEB was injected each time OVA was applied to the skin (eight times) . SEB (but not SEA) altered the development of immediate hypersensitivity to OVA, as demonstrated by the reduction in allergen-specific IgE, decreased OVA-specific immediate skin test responsiveness, and prevented the development of increased airways responsiveness after bronchial challenge with OVA . Injections of SEB did not alter the proliferative responses of local draining lymph node cells or spleen mononuclear cells to OVA, indicating that administration of SEB did not inhibit the sensitization of OVA, but shifted the immune response away from an immediate type response (IgE/IgG1) to IgG2a, IgG2b and IgG3 . Although both protocols of SEB treatment did not lead to a major deletion of the V beta 8 T cell population, they did reduce the proliferative response of V beta 8+ T cells to OVA . These data indicate that the bacterial toxin SEB is capable of modifying the immediate hypersensitivity response induced by OVA by altering the functional capacity of antigen-reactive V beta 8 T cells.

J Lab Clin Med, 1994 Dec, 124(6), 802 - 7
Bacterial characterization in Staphylococcus epidermidis septicemia; Dall L et al.; An analysis of the in vitro characteristics of Staphylococcus epidermidis strains isolated from patients with true S . epidermidis septicemia was undertaken . From a potential population of 921 cultures from adult patients with coagulase-negative bacteremia, highly defined selective criteria limited the population to 20 patients with S . epidermidis sepsis, from whose blood cultures the study organisms were isolated . Another population of 11 S . epidermidis blood isolates, clinically determined to be contaminants, were tested as a control group . In vitro assays performed on all isolates included slime quantification, hydrophobicity, surface hexoses, and capsule presence . Murine spleen phagocytosis of intravenously administered isolates was measured in vivo . The assayed quantity of cell-associated bacterial hexose sugars positively correlated with organism virulence to the host (p = 0.02) . This bacterial population was also low in slime but varied as to the presence of capsule and ease of phagocytosis . Permanent catheter-bearing patients' bacteria were somewhat more hydrophobic (p = 0.07) . We conclude that in vitro assays can differentiate bacteremic cultures from contaminants and that the characteristic that best relates to host toxicity in these S . epidermidis isolates was bacterial cell surface-associated carbohydrate.

Bioorg Med Chem, 1994 Dec, 2(12), 1413 - 5
Phage display of catalytically active staphylococcal nuclease; Ku J et al.; Staphylococcal nuclease (SNase), a 14 kD enzyme that catalyzes the hydrolysis of single- and double-stranded nucleic acid, was fused to the N-terminus of the gene III (pIII) protein of filamentous phage fdtet . The SNase-pIII protein is infective and the catalyzes DNA hydrolysis, demonstrating that functional SNase can be displayed on the phage surface.

Protein Sci, 1994 Dec, 3(12), 2175 - 84
Thermodynamic characterization of an equilibrium folding intermediate of staphylococcal nuclease; Xie D et al.; High-sensitivity differential scanning calorimetry and CD spectroscopy have been used to probe the structural stability and measure the folding/unfolding thermodynamics of a Pro117-->Gly variant of staphylococcal nuclease . It is shown that at neutral pH the thermal denaturation of this protein is well accounted for by a 2-state mechanism and that the thermally denatured state is a fully hydrated unfolded polypeptide . At pH 3.5, thermal denaturation results in a compact denatured state in which most, if not all, of the helical structure is missing and the beta subdomain apparently remains largely intact . At pH 3.0, no thermal transition is observed and the molecule exists in the compact denatured state within the 0-100 degrees C temperature interval . At high salt concentration and pH 3.5, the thermal unfolding transition exhibits 2 cooperative peaks in the heat capacity function, the first one corresponding to the transition from the native to the intermediate state and the second one to the transition from the intermediate to the unfolded state . As is the case with other proteins, the enthalpy of the intermediate is higher than that of the unfolded state at low temperatures, indicating that, under those conditions, its stabilization must be of an entropic origin . The folding intermediate has been modeled by structural thermodynamic calculations . Structure-based thermodynamic calculations also predict that the most probable intermediate is one in which the beta subdomain is essentially intact and the rest of the molecule unfolded, in agreement with the experimental data . The structural features of the equilibrium intermediate are similar to those of a kinetic intermediate previously characterized by hydrogen exchange and NMR spectroscopy.

Behring Inst Mitt, 1994 Dec, (95), 80 - 4
Functionally inactive S . aureus alpha-toxin containing a single amino acid substitution: potential usefulness as a vaccine; Bhakdi S et al.; Substitution of His-35 with arginine in staphylococcal alpha-toxin leads to loss of its membrane perturbating properties in vitro . In this study, we report that the inactive mutant toxin is also devoid of toxic properties when injected intravenously into rabbits . Whereas a single application of native toxin at a dose of 20 micrograms/kg proved uniformly lethal within 3 hours (n = 3), all animals receiving 20, 100 or 200 micrograms/kg mutant toxin (n = 3 for each group) remained in unaltered, healthy states over a 15 day period of observation . Furthermore, all animals receiving inactive toxin developed antibody titers on day 10-15 . No dose dependency was noted in the tested range . Thus, the site-directed alpha-toxin mutant (His-35:Arg) is an excellent vaccine candidate.

Med Hypotheses, 1994 Dec, 43(6), 361 - 71
CD4 similarity to proteins of infectious agents in AIDS and their role in autoimmunity; Root-Bernstein RS et al.; Lymphocytotoxic autoimmunity (LA) is ubiquitous in AIDS . Its causes are unknown . We report that significant amino acid sequence similarities exist between the proteins of infectious organisms associated with AIDS and the CD4 protein of T-helper lymphocytes . These included: HIV, cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex viruses (HSV), Varicella Zoster virus (VZV), Escherichia coli, Mycobacteria, Mycoplasmas, Plasmodium, and Staphylococcus . It has been reported previously that HIV proteins have significant similarities with human class II MHC (HLA class II) proteins . Since CD4 and HLA class II proteins are chemically complementary, pairs of homologous antigens will also be complementary . It follows that concurrent infections with CD4 and HLA class II-homologous antigens will result in idiotype-antiidiotype antibody pairs that cannot distinguish 'self' from 'nonself', that acts as lymphocytotoxins, and form circulating immune complexes . Thus, combined HIV-CMV, HIV-EBV, HIV-HBV, HIV-mycoplasma, or other appropriate infectious pairs may suffice to trigger LA in AIDS.

Biophys J, 1994 Dec, 67(6), 2511 - 21
Resolution of multiphasic reactions by the combination of fluorescence total-intensity and anisotropy stopped-flow kinetic experiments; Otto MR et al.; Multiphasic kinetics are often observed in stopped-flow investigations . To characterize further these kinetic phases, we have developed a methodology whereby fluorescence total intensity and anisotropy stopped-flow data can be combined in a single analysis . Fluorescence total intensity and anisotropy are highly interrelated and contain two very complementary forms of information . Total-intensity changes are useful in determining changes in populations with differing quantum yields, whereas anisotropy changes contain additional contributions caused by the rotational dynamics of the species . For cases in which the fluorescence quantum yield increases, the observed rate of anisotropy change will be more rapid than the total-intensity change, whereas in cases in which the total intensity decreases, the observed change in anisotropy will lag behind . In all cases, with quantum yield changes the stopped-flow anisotropy signals cannot be fit with models consisting of exponentials . Case studies examining these effects are described for the protein folding/refolding transitions of Staphylococcal nuclease and phosphoglycerate kinase . A multiphasic DNA exonuclease reaction using bacteriophage T4 DNA polymerase is also examined . In all of these cases, combined analysis of both data types revealed insights into reaction mechanism, which could not be obtained by either data type in isolation . Quantum yields and steady-state anisotropies associated with transiently populated intermediate species can be resolved . The data analysis methodologies described allow characterization of multiphasic reactions in terms of internally consistent kinetic rates, quantum yields, and steady-state anisotropies.

Int Immunol, 1994 Dec, 6(12), 1967 - 76
Activation-induced expression of thymic shared antigen-1 on T lymphocytes and its inhibitory role for TCR-mediated IL-2 production; Kosugi A et al.; We have produced a hamster mAb, PRST1, which reacts with thymic shared Ag-1 (TSA-1), a product of the Ly6 gene family . By cross-blocking experiments, we found that TSA-1 is identical to stem cell Ag-2 (Sca-2) . Using PRST1, the changes of TSA-1/Sca-2 expression on mature T cells during the activation process were analyzed . Although freshly isolated T cells did not express detectable TSA-1 on their cell surface, in vitro stimulation of T cells with concanavalin A induced a marked increase of surface TSA-1 expression . The increased expression of TSA-1 on T cells was detected from 12 h after stimulation and was associated with the increase of TSA-1 mRNA . In vivo injection of mice with staphylococcal enterotoxin B (SEB) resulted in the enhanced TSA-1 expression in splenic V beta 8+ T cells . This antigen-specific induction of TSA-1 expression in vivo preceded a detectable increase in numbers of V beta 8- T cells after SEB injection . Functionally, whereas anti-TSA-1 mAb was not mitogenic to T cells, it inhibited anti-CD3-induced IL-2 production by T cell hybridomas . These results indicate that TSA-1/Sca-2 is a unique marker for T cell activation and a signal through this molecule may have a negative feedback role to limit IL-2 production from activated T cells stimulated through the TCR.

Int Immunol, 1994 Dec, 6(12), 1865 - 74
Monoclonal antibodies to a TCR idiotope modulate the superantigen-induced responses of encephalitogenic rat T cells; Zhou SR et al.; Superantigens, such as staphylococcal enterotoxins, activate T lymphocytes by linking MHC class II molecules on antigen presenting cells to the V beta element of the TCR . Through this effect on T cells, superantigens may influence the immune response and autoimmune disease . In fact, superantigens may activate or anergize cells involved in the production of experimental allergic encephalomyelitis . Lewis rats recognize an encephalitogenic epitope in myelin basic protein (MBP) residues 68-88 through an MHC class II I-A restricted process using TCR V beta 8.2 . The F30 murine mAb reacts with an encephalitogenic idiotope (Id) on the TCR of V beta 8.2+ encephalitogenic Lewis rat T cells . In the present study it was demonstrated that the same mAb anti-Id inhibited the proliferation and IL-2 secretion induced by staphylococcal enterotoxins A, B and E in a V beta 8.2+ encephalitogenic Lewis rat T cell line specific for guinea pig MBP peptide 68-88 . The mAb anti-Id did not inhibit the response of control T cells similarly derived but inhibited V beta 8.2- and recognizing MBP peptide 87-99 . Control anti-Id failed to inhibit the response of either cell line . These findings imply that the specific antigen and superantigen react with TCR in a manner similar enough to be inhibited by the same anti-Id . The mechanism may involve the induction of anergy by the anti-Id, interference/steric hindrance by the reaction of anti-Id with TCR and possibly a little direct reaction of anti-Id with superantigens . Anti-Id directed immunotherapy may have a role in modulating the damage of inflammatory demyelination induced by both specific antigens and superantigens.

Zentralbl Veterinarmed B, 1994 Dec, 41(10), 639 - 44
Double-sandwich enzyme-linked immunosorbent assay (ELISA) with murine monoclonal antibodies for detection of staphylococcal enterotoxin B; Goyache J et al.; Three monoclonal antibodies (MAbs) which react specifically with staphylococcal enterotoxin B (SEB) were studied for their suitability for use in ELISA . One of them (MAb B14) worked well as a coating antibody; MAb B12 was shown to be a good probing antibody (conjugated with peroxidase) when ELISA plates were coated with MAb B14 . This effective pair of MAbs (B14-B12PO) is able to detect 0.625 ng SEB ml, and to distinguish between SEB and other proteins present in food extracts.

Int Immunol, 1994 Dec, 6(12), 1849 - 56
Involvement of apoptosis antigen Fas in clonal deletion of human thymocytes; Yonehara S et al.; Apoptosis appears to play a major role in the differentiation and selection of T and B lymphocytes, but the mechanisms of clonal deletion of T cells in thymus are not well understood . We have prepared an anti-human Fas IgM mAb with associated apoptosis-inducing activity in Fas antigen-positive target cells including human T cells . We analyzed the expression of apoptosis antigen Fas on human thymocytes by cytofluorometry showing low, but significant amounts of Fas antigen on double-negative and double-positive undifferentiated thymocytes . On the contrary, most of the differentiated thymocytes (single-positive or CD3-brightest) expressed undetectable levels of Fas antigen . About 1-2% of thymocytes expressed high amounts of Fas antigen, and these cells, which were CD3-bright, were CD4-bright and CD8-low at the stage of late double-positive lineage . Immunohistological analysis shows these Fas-bright cells on the edge of the medulla . Stimulation through the TCR complex was shown to induce the expression of Fas antigen on thymocytes at the late double-positive stage and prolonged stimulation through the TCR complex rendered the Fas-bright thymocytes sensitive to apoptosis-inducing activity of anti-Fas . To show the involvement of the Fas system in the negative selection/clonal deletion of thymocytes, we organ-cultured human thymus in the presence of the superantigen, staphylococcus enterotoxin B (SEB), and new antagonistic anti-Fas mAb, which can inhibit the apoptosis-inducing activity of the original anti-Fas mAb . The SEB-reactive TCR complex on thymocytes was at first down-regulated by SEB, then the SEB-reactive clone was deleted by apoptosis, which was inhibited by an antagonistic anti-Fas mAb . Thus, Fas antigen is shown to be involved in the negative selection/clonal deletion of superantigen-reactive thymocytes.

J Autoimmun, 1994 Dec, 7(6), 697 - 710
Delayed kinetics of T lymphocyte anergy and deletion in lpr mice; Mixter PF et al.; The Fas/APO-1 (Fas) antigen is a cell surface protein that mediates apoptosis and belongs to the tumor necrosis factor receptor family . The lymphoproliferative (lpr) anomaly in mice results from a retroviral disruption within the fas gene . Mice that are homozygous for the lpr mutation accumulate large numbers of T lymphocytes and exhibit an autoimmune syndrome resembling systemic lupus erythematosus . A possible explanation for this process is that in the absence of Fas antigen, lpr T cells may be resistant to normal peripheral deletional signals . The bacterial superantigen staphylococcal enterotoxin B (SEB) rapidly induces anergy and deletion by apoptosis of reactive T lymphocytes in normal mice . Administration of SEB to adult lpr mice results in the delayed induction of both unresponsiveness and deletion of V beta 8+ lymph node cells . This is not due merely to an increased thymic output in lpr mice; the delayed induction of tolerance and elimination of reactive lpr T cells by superantigens are intrinsic properties of the cells . The progressive lymphadenopathy in lpr mice may reflect a process of lymphoaccumulation rather than lymphoproliferation . A delay in tolerance induction and elimination of self-reactive T cells could have profound influence on the autoimmune diathesis of lpr mice.

J Biol Chem, 1994 Nov 25, 269(47), 29998 - 30004
Bacterial multidrug resistance is due to a single membrane protein which functions as a drug pump; Grinius LL et al.; Multidrug transport system in proteoliposomes was reconstituted using the highly purified membrane transport protein responsible for bacterial multidrug resistance . This protein (named Smr, for staphylococcal multidrug resistance) consists of 107 amino acid residues and displays four putative transmembrane domains . The Smr protein was tagged with a FLAG epitope, and the modified protein was expressed, purified, characterized, and reconstituted into proteoliposomes . With this in vitro experimental system, it has been demonstrated that a highly purified multidrug resistance protein functions as a drug pump, which transports methyltriphenylphosphonium actively against a 10(3)-fold concentration gradient . Delta mu H+ was shown to be a driving force, and an electrogenic drug/proton antiport was suggested as the molecular mechanism of the drug transport . Of the 2 Glu residues in putative extramembrane loops of the Smr polypeptide chain, Glu-24 was shown to be involved in determining the specificity of drug resistance . Replacement of both of these Glu residues with Asp produced active Smr . In contrast, Smr was unable to protect cells from multiple drugs when a Glu-13-->Asp-13 replacement was made . We suggest that Glu-13, a unique acidic residue located in the hydrophobic domain of Smr, is directly involved in the drug/proton antiport.

Biochemistry, 1994 Nov 22, 33(46), 13625 - 41
Mapping staphylococcal nuclease conformation using an EDTA-Fe derivative attached to genetically engineered cysteine residues; Ermacora MR et al.; Six single cysteine variants of staphylococcal nuclease were reacted with the iron complex of (EDTA-2-aminoethyl) 2-pyridyl disulfide (EPD-Fe) {Ermacora, M . R., Delfino, J . M., Cuenoud, B., Schepartz, A., & Fox, R . O . (1992) Proc . Natl . Acad . Sci . U.S.A . 89, 6383-6387} and used to assess the ability of this cleavage reagent to faithfully report on the structure of nonnative protein states . The act of mutation and modification did not significantly alter the protein's global structure, as measured by CD and enzymatic activity, and only modestly affected its stability . The reaction was conformation dependent and generated specific cleavage products that mapped tertiary interactions present in the folded state . Several parameters relevant to the cleavage reaction and its use as a conformational probe were analyzed . Proximity and solvent accessibility are the most important parameters in determining the cleavage pattern and can be used to predict cleavage sites in the native protein . The cleavage reaction can be performed in the presence of high denaturant concentration, in the presence of SDS, and under a wide range of pH values; thus it can readily be applied to the study of equilibrium folding intermediates . Mass spectrometric analysis combined with N-terminal sequencing identified cleavage products consistent with a single cleavage event per protein molecule and revealed one cleavage mechanism which was not previously considered for protein oxidative degradation, although it was reported for hydroxyl radical induced cleavage of small peptides . Identification of the cleavage sites obtained from each variant allowed a nearest-neighbor mapping of the secondary structural elements of nuclease . Quantitation of specific cleavage products was used to monitor the disruption of the interaction between helices H2 and H3 in equilibrium unfolding experiments . The resulting unfolding curve revealed a local conformational heterogeneity at low denaturant concentration which was not observed when the same transition was monitored by the change in fluorescence of a single nuclease tryptophan . Interestingly, the midpoint of the transition and the second half of the unfolding curve were the same, as monitored by the two probes . This indicates that the lifetime of the reactive oxygen species generated by the cleavage reagent is short compared to the unfolding equilibrium rate constants and that the cleavage technique identifies a native-like folding intermediate not detected by fluorescence . The experiments presented herein demonstrate that EPD-Fe-mediated protein cleavage is an appropriate technique for the study of nonnative protein structure.(ABSTRACT TRUNCATED AT 400 WORDS)

Orv Hetil, 1994 Nov 20, 135(47), 2597 - 602
{CT-guided percutaneous drainage in the management of psoas abscesses}; Horvath G et al.; The authors review their first experiences in Hungary with the CT guided percutaneous catheter treatment of psoas abscesses . Diagnostic aspiration from abscess was performed in one patient . Percutaneous catheter drainage of psoas abscess were performed in 4 patients . There were 4 patients with unilateral and 1 patient with bilateral psoas abscess . 4 patients had tuberculous spondylodiscitis, 1 patient had staphylococcal vertebral osteomyelitis . One patient following diagnostic aspiration was successfully treated with antituberculoutic drugs . All patient treated with percutaneous catheter drainage received medical antituberculoutic or antibiotic therapy and showed excellent clinical and radiological results . There was no complication and there was no need to open surgical drainage in this series . CT guided aspiration is a valuable tool in the diagnosis of psoas abscesses as well as in the identification of the exact abscess etiology . Percutaneous drainage represents an efficient and attractive alternative to open surgical drainage in the treatment of psoas abscesses.

Biochem Biophys Res Commun, 1994 Nov 15, 204(3), 1212 - 8
The two subunits of a phospholipase A2 inhibitor from the plasma of Thailand cobra having structural similarity to urokinase-type plasminogen activator receptor and LY-6 related proteins; Ohkura N et al.; Amino acid sequences of the two subunits (31-kDa and 25-kDa subunits) of a phospholipase A2 (PLA2) inhibitory protein, purified from the blood plasma of Thailand cobra Naja naja kaouthia, were determined by alignment of peptides obtained by lysyl endopeptidase, staphylococcal V8 protease and endoproteinase Asp-N digestions . The respective subunits were composed of 188 and 185 amino acid residues, and the former contained one asparagine-linked sugar chain at the position 157 . There was 29% identity between 31-kDa and 25-kDa subunits . The analysis of internal homology in each sequence of the two subunits revealed the existence of two repeats of approximately 90 amino acid residues . These sequence units were found to be significantly homologous to those of urokinase-type plasminogen activator receptor and of Ly-6 related proteins, such as Ly-6A/E, Ly-6C, ThB, and CD59 antigens.

Schweiz Med Wochenschr, 1994 Nov 12, 124(45), 2060 - 3
{Type and frequency of severe infectious complications in the treatment of acute leukemia . Prospectively acquired findings in a consecutive patient cohort}; Jud B et al.; 139 patients with newly diagnosed acute leukemia were admitted to our institution from 1984 to 1990 . More than 70% developed severe infections during cytotoxic chemotherapy . Septicemia accounted for 40.1% . The two most frequent organisms isolated from the blood were Staphylococcus epidermidis and E . coli . A prominent finding were the soft tissue infections (23.4%) . They often occurred in the anorectal area and at the exit of an intravenous catheter . Pneumonia (17.8%) was of fungal origin in 22.9% . 66% of the infections occurred when the neutrophils were below 100/microliters.

J Chromatogr A, 1994 Nov 11, 685(1), 31 - 7
Resolution of proteins on a phenyl-Superose HR5/5 column and its application to examining the conformation homogeneity of refolded recombinant staphylococcal nuclease; Jing G et al.; In order to examine the effect of amino acid substitutions on protein retention in hydrophobic interaction chromatography and the resolution of a phenyl-Superose HR5/5 column, two groups of staphylococcal nucleases, named Y113/W140 (wild-type), Y113W/W140 and Y113/W140F, Y113W/W140F, were produced by substituting tryptophan (W) for tyrosine (Y) at residue 113 and phenylalanine (F) for tryptophan (W) at residue 140 . For each group, the proteins have the same amino acid at residue 140, but a different amino acid at residue 113 . The solvent perturbation of nuclease fluorescence and 1,8-anilinoaphthalene-8-sulfonate binding studies showed that the substitutions do not change the side-chain positions of amino acids at residues 113 and 140 . Chromatography of the proteins on the Phenyl-Superose HR5/5 column showed that the proteins with tryptophan at residue 113 have longer retention times than the proteins having tyrosine at residue 113; the proteins with the same amino acid at residue 113 have almost the same retention time regardless of substituting phenylalanine for tryptophan at residue 140 . The studies clearly indicate that not all amino acid substitutions have an effect on protein retention; the contribution to retention of a given amino acid substitution depends on its position in a protein . Single amino acid substitutions at the exterior surface of a protein, which change the strength of hydrophobic interaction, can affect the protein retention in hydrophobic interaction chromatography . Staphylococcal nuclease and its mutants with only one amino acid difference on their surfaces can be discriminated by the phenyl-Superose column.(ABSTRACT TRUNCATED AT 250 WORDS)

J Exp Med, 1994 Nov 1, 180(5), 1931 - 5
Evidence for a functional interaction between the beta chain of major histocompatibility complex class II and the T cell receptor alpha chain during recognition of a bacterial superantigen; Deckhut AM et al.; Several studies have suggested that there is a direct interaction between the T cell receptor (TCR) and the major histocompatibility complex (MHC) molecule during T cell recognition of superantigen . To further investigate this possibility, we have analyzed T cell recognition of a bacterial superantigen, Staphylococcal enterotoxin B (SEB), presented by a series of mutant murine I-Ek molecules in which residues of either the alpha or beta chain predicted to interact with the TCR have been substituted . Individual T cell hybridomas gave distinct patterns of responsiveness to SEB presented by the I-E beta k mutants that could not be attributed to differences in the binding of SEB to the mutants . This effect appeared to be dependent on the TCR-alpha chain because some of these hybridomas expressed identical TCR transgenic beta chains . In contrast, none of the hybridomas gave distinct patterns of responsiveness to SEB presented by the I-E alpha k mutants . Taken together, these observations support the idea that there is a functional interaction between the alpha chain of the TCR and the beta chain of the MHC class II molecule . The data also support the idea that this interaction might enhance superantigen recognition in some cases.

J Exp Med, 1994 Nov 1, 180(5), 1921 - 9
T cell receptor-major histocompatibility complex class II interaction is required for the T cell response to bacterial superantigens; Labrecque N et al.; Bacterial and retroviral superantigens (SAGs) stimulate a high proportion of T cells expressing specific variable regions of the T cell receptor (TCR) beta chain . Although most alleles and isotypes bind SAGs, polymorphisms of major histocompatibility complex (MHC) class II molecules affect their presentation to T cells . This observation has raised the possibility that a TCR-MHC class II interaction can occur during this recognition process . To address the importance of such interactions during SAG presentation, we have used a panel of murine T cell hybridomas that respond to the bacterial SAG Staphylococcal enterotoxin B (SEB) and to the retroviral SAG Mtv-7 when presented by antigen-presenting cells (APCs) expressing HLA-DR1 . Amino acid substitutions of the putative TCR contact residues 59, 64, 66, 77, and 81 on the DR1 beta chain showed that these amino acids are critical for recognition of the SAG SEB by T cells . TCR-MHC class II interactions are thus required for T cell recognition of SAG . Moreover, Mtv-7 SAG recognition by the same T cell hybridomas was not affected by these mutations, suggesting that the topology of the TCR-MHC class II-SAG trimolecular complex could be different from one TCR to another and from one SAG to another.

J Infect Dis, 1994 Nov, 170(5), 1189 - 94
Superantigen shock in mice with an inapparent viral infection; Sarawar SR et al.; Subclinical lymphocytic choriomeningitis virus infection primes mice expressing a V beta 8.1D beta 2J beta 2.3C beta 2 T cell receptor as a transgene for induction of fatal hematogenous shock after administration of a dose of staphylococcal enterotoxin B (SEB) that is tolerated by uninfected controls . The lethal effect is greatly diminished by prior depletion of the virus-primed CD4+ T cells . Evidence of transient tumor necrosis factor (TNF) secretion is detected in serum within 1 h of SEB administration, and massive amounts of interferon-gamma (IFN-gamma) and interleukin-6 (IL-6) are present within 4-6 h . Mice are partly protected by treatment with dimeric soluble TNF receptor-Fc fusion protein or the nitric oxide synthase inhibitor, aminoguanidine, neither of which blocks SEB-induced IFN-gamma or IL-6 production . Administration of a monoclonal antibody to IFN-gamma concomitant with SEB effectively neutralizes this cytokine but has no effect on survival.

J Immunol, 1994 Nov 1, 153(9), 4247 - 55
Apoptotic cell death induced by intracellular proteolysis; Williams MS et al.; To mimic the injection of granzymes into target cells by cytotoxic lymphocytes or the activation of endogenous proteases in programmed cell death, the proteases chymotrypsin, proteinase K, or trypsin were loaded into the cytoplasm of several different cell types using the osmotic lysis of pinosomes technique . Internalization of these proteases caused cell lysis within several hours, accompanied by extensive nuclear damage in most but not all combinations of target cells and proteases . This nuclear damage, quantitated by DNA release from nuclei, was associated with apoptotic features including DNA fragmentation into nucleosomal ladders, chromatin condensation, nuclear fragmentation, and membrane blebbing . Agents reported to block programmed cell death, including aurintricarboxylic acid, inhibitors of energy metabolism, and protein or RNA synthesis, failed to block this protease-induced death, although some inhibited nuclear damage . In separate experiments, introduction of staphylococcal nuclease into cells led to near complete (at least 75% of total) nucleosomal DNA fragmentation within 6 to 8 h . Condensation of chromatin did not accompany this fragmentation to the same extent, and there was approximately a 10-h lag between half-maximal DNA fragmentation and 50% loss of membrane integrity . The results suggest that activation of intracellular proteases during cell death by any molecular pathway could give rise to apoptotic morphology and DNA fragmentation.

J Immunol, 1994 Nov 1, 153(9), 3908 - 16
Immunosuppressant deoxyspergualin inhibits antigen processing in monocytes; Hoeger PH et al.; Deoxyspergualin (DSG) is a novel immunosuppressive agent recently shown to bind to the constitutive heat shock protein 70, which is involved in binding and intracellular transport of antigenic peptides . In this study, we show that DSG inhibits the proliferation of PBMCs to the Ags tetanus toxoid and diphtheria toxoid, but not to the mitogens PHA and PMA/ionomycin, nor to the superantigens toxic shock syndrome toxin-1 and staphylococcal enterotoxin A . DSG's effect was specific for monocytes as preincubation of T cells with DSG did not inhibit their proliferation to monocytes pulsed with tetanus toxoid Ag for 16 h, whereas the presence of DSG during Ag pulsing of the monocytes inhibited their ability to stimulate T cell proliferation . DSG did not down-regulate the expression of MHC class II molecules by monocytes, and the inhibitory effect of DSG on T cell proliferation was not reversed by the addition of IL-2, nor by the addition of the costimulatory signals IL-1, IL-6, and anti-CD28 . Studies with two human T cell clones, HA1.7 and PF5, specific, respectively, to peptides spanning amino acids 307-319 and 256-270 of influenza hemagglutinin, showed that DSG inhibited the proliferation of the clones to the native hemagglutinin molecule but minimally affected their proliferation to the peptides . These data suggest that DSG interferes with Ag processing and/or presentation.

Infect Immun, 1994 Nov, 62(11), 4775 - 80
Localization of binding sites of staphylococcal enterotoxin B (SEB), a superantigen, for HLA-DR by inhibition with synthetic peptides of SEB; Komisar JL et al.; Staphylococcal enterotoxins are major causes of food poisoning and toxic shock syndrome . Their ability to bind to major histocompatibility complex (MHC) class II molecules has been suggested to be the first step in the mechanism whereby they cause illness . By flow cytometric analysis, the sites of interaction of staphylococcal enterotoxin B (SEB) with HLA-DR molecules were probed in the present study by inhibiting the binding of biotinylated SEB to a human T-cell line (HUT-78) with synthetic peptides of SEB . Five peptides of SEB gave significant inhibition of binding: a peptide containing amino acids 9 to 20 {SEB(9-20)}, SEB(30-38), SEB(61-70), SEB(90-114), and SEB(169-181) . One peptide, SEB(39-51), enhanced binding . Among the inhibitory peptides, SEB(90-114), a peptide spanning the entire disulfide loop, showed the most efficient inhibition of binding . Peptides SEB(9-20) and SEB(39-51) include amino acid residues that have been identified by previous mutation studies (J.W . Kappler, A . Herman, J . Clements, and P . Marrack, J . Exp . Med . 175:387-396, 1992) as being important in binding to MHC class II . Amino acids lining the alpha 5 groove of SEB have also been postulated to be involved in binding to MHC class II molecules . However, only two of the residues that line the alpha 5 groove of SEB, His-12 and Tyr-17, are on peptide SEB(9-20) that inhibits binding . These results confirm previous studies that implicated the amino-terminal portion of the molecule in binding to MHC class II molecules and further indicate an important role for residues in other regions, particularly the disulfide loop.

J Clin Immunol, 1994 Nov, 14(6), 340 - 52
Acute ethanol consumption synergizes with trauma to increase monocyte tumor necrosis factor alpha production late postinjury; Szabo G et al.; The hypothesis that acute ethanol uptake plus trauma can synergize to increase immunosuppression was tested . We found that, unlike non-alcohol-exposed patients, patients with acute alcohol use prior to trauma have a transient decrease in monocyte tumor necrosis factor alpha (TNF alpha) production during the very early postinjury (0-3 days) period . However, TNF alpha production by these alcohol-exposed patients' monocytes (M0) became hyperelevated late postinjury (> 9 days) . Consequently, these massively elevated M0 TNF alpha levels can contribute to posttrauma immunosuppression after acute alcohol use . We also demonstrate that normal monocyte activation with the superantigen, Staphylococcus enterotoxin B (SEB), results in a preferential induction of cell-associated M0 TNF alpha production, described as characteristic of immunosuppressed trauma patients . Acute in vitro ethanol treatment down-regulated the elevated TNF alpha production by trauma patients' M0 after either SEB, muramyl-dipeptide (MDP), interferon-gamma plus MDP, or lipopolysaccharide (LPS) stimulation . Both SEB- and LPS-induced TNF alpha mRNA induction was inhibited by acute alcohol treatment in normal M0, indicating that ethanol can regulate cytokine gene expression . An additional immunosuppressive effect of acute ethanol's stimulation was suggested by its induction of elevated transforming growth factor beta production in trauma patients' activated M0.

Can Vet J, 1994 Nov, 35(11), 699 - 701
Bacterial and fungal flora in healthy eyes of birds of prey; Dupont C et al.; Birds of prey are often affected with external ocular injuries that are routinely treated with antimicrobial agents used for small animals . The resident ocular bacterial and fungal flora is still unknown in birds of prey and this knowledge would be very useful in assessing the accuracy of treatments . In a study involving 65 raptors with healthy eyes, swabs were taken from both eyes to identify the resident bacterial and fungal flora . Fifty-five birds had a positive culture in one or both eyes . Both gram-positive and gram-negative organisms were isolated, with a predominance of Staphylococcus spp., which were found in 52.3% of cultures . Only two fungal species, Aspergillus spp . and Cladosporium spp . were found . The overall results of this study are similar to previous studies carried out in humans and other animals.

Jpn J Antibiot, 1994 Nov, 47(11), 1582 - 8
{Clinical studies on cefozopran in pediatrics}; Sekiguchi T et al.; Cefozopran (CZOP, SCE-2787) was given intravenously to 12 children with acute bacterial infections including 9 with acute pneumonia, 1 each with acute pyothorax, impetigo and staphylococcal scalded skin syndrome . Good or excellent clinical responses were obtained in all of the 12 patients and bacterial eradications were achieved for all 10 strains identified in these cases . No side effects were noted . Eosinophilia was observed in one case, however . From the above clinical results, it appears that CZOP is a useful antibiotic for treatment of pediatric patients with various bacterial infections.

Nippon Kyobu Geka Gakkai Zasshi, 1994 Nov, 42(11), 2081 - 6
{A case of left ventricular-right atrial communication complicated with aortic regurgitation caused by active infective endocarditis}; Okada Y et al.; A 63-year-old man with cough and shortness of breath was diagnosed to have active infective endocarditis resulting in acute pulmonary edema with aortic regurgitation caused by a huge vegetation on the noncoronary cusp and left ventricular-right atrial (LV-RA) communication which were demonstrated by the echocardiogram and color doppler method . The LV-RA communication located at the atrioventricular portion of the membranous septum was closed with GoreTex patch through the right atrium combined with the aortic valve replacement with a bileaflet mechanical valve in emergency . The bacteriological studies demonstrated staphylococcus epidermidis . The postoperative course was uneventful and the patient is now on regular duty two years after surgery.

Immunology, 1994 Nov, 83(3), 333 - 40
Superantigen-induced anergy of V beta 8+ CD4+ T cells induces functional but non-proliferative T cells in vivo; Gaus H et al.; The response profile of staphylococcal enterotoxin B (SEB)-primed murine V beta 8+ CD4+ and V beta 8+ CD8+ T cells was analysed upon rechallenge in vitro . While in vitro responses to secondary stimulation with SEB were reduced to background levels, the in vivo reactivity after rechallenge with SEB was retained, in that SEB-primed mice succumbed to lethal T-cell shock, lymphokines {interleukin-1 (IL-1), IL-2, Il-4, IL-6, IL-10, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha)}, and lymphokine-specific mRNA accumulation could be detected in V beta 8+ CD4+ and V beta 8+ CD8+ T cells . However, V beta 8+ CD4+ T cells failed to enter the cell cycle . While the phenotype of V beta 8+ CD8+ T cells was indistinguishable from that of their counterparts from naive mice, V beta 8+ CD4+ T cells exhibited in vivo an unusual phenotype as non-proliferative but functional T cells . We conclude that in vitro-defined anergy does not disclose the functional abilities of ligand-reactive V beta 8+ T cells in vivo, and that priming with superantigen (SAg) induces in vivo a differentiation of SEB-reactive V beta 8+ CD4+ T cells into a non-proliferative but functional phenotype.

Prikl Biokhim Mikrobiol, 1994 Nov-Dec, 30(6), 931 - 9
{Solid phase methods of immunoenzyme analysis of the herbicides 2,4-dichlorophenoxyacetic and 2,4,5-trichlorophenoxyacetic acids}; Dzantiev BB et al.; Competitive solid-phase enzyme immunoassays for the detection of 2,4-dichlorophenoxyacetic (2,4-D) and 2,4,5-trichlorophenoxyacetic (2,4,5-T) acids have been developed and optimized . The sensitivity of the assay is 3 ng/ml for 2,4-D and 5 ng/ml for 2,4,5-T . The time of the assay is 1.5 h . The sensitivity of the assays increases after immobilization of antibodies on staphylococcus protein A, in the case of using monovalent derivatives of antibodies, and as a result of chemical modification of hapten . The detection limit for the pesticides is 0.1 ng/ml in this case, which is close to the sensitivity of the assay for the haptens conjugated with proteins.

Arch Mal Coeur Vaiss, 1994 Nov, 87(11), 1453 - 8
{Results, complications and long-term follow-up of percutaneous ablation of atrioventricular conduction . Apropos of 85 cases}; Sadoul N et al.; This retrospective study reports the immediate and long-term results of percutaneous ablation of atrioventricular conduction . Between July 1983 and January 1992, 85 consecutive patients (51 men, age 64 +/- 10 years, range 43-84 years) presenting with supraventricular arrhythmias (atrial fibrillation n = 53; atrial flutter n = 50; atrial tachycardia n = 17; junctional tachycardia n = 6) resistant to antiarrhythmic therapy (number of drugs used: 4 +/- 1.3, range 1-6) underwent interruption of atrioventricular conduction by fulguration (n = 65) or radiofrequency energy (n = 13) or by an association of the two methods (n = 7) . The 75 pacemakers implanted (10 patients had pacemakers before the procedure) comprised 55 VVIR, 11 VVI, 5 DDD and 4 DDDR units . The immediate results included two sudden deaths at the 4th and 7th day in patients undergoing fulguration and three complications with a favourable outcome (staphylococcal septicaemia, pulmonary embolism and haematoma at the site of implantation of the pacemaker) . None of the patients was lost to follow-up and the average follow-up was 31 +/- 18 months (range 2-108 months) . During follow-up, 15 patients died and there was a recurrence of symptoms in 11 patients after 1 to 9 months requiring a repeat procedure . In the 68 survivors, the follow-up is now 38 +/- 18 months (range 12-108 months) . Sixty one patients have 2nd (2) or 3rd (59) degree atrioventricular block, giving 90% good electrocardiographic results.(ABSTRACT TRUNCATED AT 250 WORDS)

Can J Physiol Pharmacol, 1994 Nov, 72(11), 1434 - 9
Protein tyrosine phosphorylation, cellular Ca2+, and Ca2+ sensitivity for contraction of smooth muscle; Di Salvo J et al.; Our studies are guided by the novel hypothesis that protein tyrosine phosphorylation is an important mechanism for regulating contraction of smooth muscle . Several lines of evidence are reviewed which suggest that enhanced tyrosine phosphorylation participates in mechanisms that regulate cytosolic Ca2+ and Ca2+ sensitivity for contraction . First, vanadate-induced contraction of guinea-pig taenia coli is functionally linked to enhanced protein tyrosine phosphorylation of at least three substrates, apparently resulting from vanadate-mediated inhibition of protein tyrosine phosphatase activity . Second, vanadate-induced contraction is dependent on extracellular Ca2+ . Third, increases in cytosolic Ca2+ resulting from stimulation of alpha 1-adrenergic receptors in cultured canine vascular smooth muscle cells are associated with enhanced tyrosine phosphorylation and are inhibited by genistein, a potent inhibitor of tyrosine kinase activity . Fourth, genistein markedly and reversibly suppresses Ca2+ sensitivity for contraction in ileal longitudinal smooth muscle permeabilized with staphylococcal alpha-toxin . Moreover, the same or similar substrates (e.g., 42-45, 70, 80-85, 95, 100, 110, 116, and 205 kDa) are tyrosine phosphorylated in response to Ca2+ or stimulation of muscarinic or alpha 1-adrenergic receptors . Collectively, these data strongly suggest that tyrosine phosphorylation is an important mechanism for regulation of smooth muscle contraction.

Int J Artif Organs, 1994 Nov, 17(11), 603 - 8
Removal of antiacetylcholine receptor antibodies by protein-A immunoadsorption in myasthenia gravis; Berta E et al.; Myasthenia Gravis is an autoimmune disease in which autoantibodies to the acetylcholine receptor interfere with neuromuscular transmission . Plasma exchange is effective in temporarily relieving the symptoms of the disease, but for repeated use the lack of selectivity and need for replacement fluids (which increases the risk of contracting viral diseases) are important drawbacks . Staphylococcal protein A, a potent ligand for immunoglobulins, that interacts negligibly with other plasma proteins, appears to be an optimal candidate for removing antiacetylcholine receptor antibodies, which are mostly IgG . We treated three patients with severe immunosuppression-resistant myasthenia gravis with protein A immunoadsorption . Neurological impairment significantly improved in all patients . After immunoadsorption of 1.5-2 plasma volumes per session, the mean percentage reductions for serum IgG and specific autoantibodies were 71% and 82% respectively . No major side effects occurred . Protein A immunoadsorption appears to be a safe, efficient and effective alternative to plasmaexchange for selected myasthenic patients requiring prolonged apheresis.

Ann Acad Med Singapore, 1994 Nov, 23(6), 914 - 6
Congenital limb reduction defects: report of two cases; Malik AS et al.; We describe two Malay male term neonates with congenital limb reduction defects . The first neonate had hypodactyly of limbs associated with micrognathia, microstomia, glossopalatine ankylosis and congenital mitral stenosis . He developed gram-negative septicaemia and died on day 14 of life . The second neonate had tetraperomelia without any other associated congenital abnormality . He developed staphylococcal skin infection which was treated conservatively . Very few cases of congenital limb reduction defects have been reported in the Asian population and we are not aware of any other reports describing Malay infants with the congenital abnormalities described in this report.

Minerva Chir, 1994 Nov, 49(11), 1107 - 9
{Ceftriaxone prophylaxis in cerebrospinal fluid shunts . Motivation and results of the choice of prophylaxis with this cephalosporin in 100 patients}; Arnaboldi L; Staphylococcus epidermidis is a bacterial organism found in almost 75% of such shunts becoming infected . This is due to its production of an extracellular slime responsible of adherence to implantable devices and resistance to antibiotic therapy . Use of antimicrobial prophylaxis prevents production of such slime . In 100 shunt operations the A . used antimicrobial prophylaxis with ceftriaxone (2 g before operation) . Follow up of 3 years showed no shunt infection . Choice of ceftriaxone was due to its pharmacokinesis.

J Biol Chem, 1994 Oct 21, 269(42), 26531 - 8
Mutually exclusive binding of peptide and invariant chain to major histocompatibility complex class II antigens; Ericson ML et al.; The invariant chain is a membrane protein associated with the major histocompatibility complex class II antigens both intra- and extracellularly . The extracellular portion of the human invariant chain (Ii) was expressed in Escherichia coli as a fusion protein with a polyhistidine tail and purified by metal affinity chromatography . The recombinant Ii was used as a ligand to probe binding to the cell surface of Chinese hamster ovary cells stably transfected with human class II alpha and beta genes of the DR4 isotype . We show that recombinant Ii inhibits peptide loading on class II polypeptides and also the converse; the presence of peptide in the antigen groove prevents binding of fluorescein-conjugated Ii . Moreover, blocking of Ii binding by peptide did not require a transition of the class II dimers to an SDS-stable state . A monoclonal antibody, L243, known to bind to (or close to) the peptide pocket of the class II molecule likewise blocked Ii-fluorescein binding . Further, we investigated whether or not the Ii, a variety of bacterial superantigens or the CD4 molecule, have overlapping binding sites on the class II heterodimer . Of the class II ligands tested, reduced binding was detected for the Staphylococcus superantigen type SEB on cells precincubated with soluble Ii while the binding of the other ligands was either unchanged or marginally changed . These data clarify by a direct biochemical approach the binding characteristics of Ii in comparison with other class II ligands.

EMBO J, 1994 Oct 17, 13(20), 4757 - 64
Structural organization of the pentameric transmembrane alpha-helices of phospholamban, a cardiac ion channel; Arkin IT et al.; Phospholamban is a 52 amino acid calcium regulatory protein found as pentamers in cardiac SR membranes . The pentamers form through interactions between its transmembrane domains, and are stable in SDS . We have employed a saturation mutagenesis approach to study the detailed interactions between the transmembrane segments, using a chimeric protein construct in which staphylococcal nuclease (a monomeric soluble protein) is fused to the N-terminus of phospholamban . The chimera forms pentamers observable in SDS-PAGE, allowing the effects of mutations upon the oligomeric association to be determined by electrophoresis . The disruptive effects of amino acid substitutions in the transmembrane domain were classified as sensitive, moderately sensitive or insensitive . Residues of the same class lined up on faces of a 3.5 amino acids/turn helical projection, allowing the construction of a model of the interacting surfaces in which the helices are associated in a left-handed pentameric coiled-coil configuration . Molecular modeling simulations (to be described elsewhere in detail) confirm that the helices readily form a left-handed coiled-coil helical bundle and have yielded molecular models for the interacting surfaces, the best of which is identical to that predicted by the mutagenesis . Residues lining the pore show considerable structural sensitivity to mutation, indicating that care must be taken in interpreting the results of mutagenesis studies of channels . The cylindrical ion pore (minimal diameter of 2 A) appears to be defined largely by hydrophobic residues (I40, L43 and I47) with only two mildly polar elements contributed by sulfurs in residues C36 and M50.

J Immunol, 1994 Oct 15, 153(8), 3611 - 21
Clonal expansion but lack of subsequent clonal deletion of bacterial superantigen-reactive T cells in murine retroviral infection; Aoki Y et al.; Several studies have suggested that activation-induced apoptosis of Ag-specific CD4+ T cells leads to depletion of this subset during HIV infection . The bacterial superantigen, staphylococcal enterotoxin A (SEA), is known to induce activation-induced apoptosis in the TCR V beta-bearing CD4+ T cells in the periphery after clonal expansion of these cells . The murine retroviral model of AIDS (MAIDS), which is induced by LP-BM5 murine leukemia virus, shares many common features with HIV infection in humans, except that CD4+ T cells increase progressively in susceptible strains . In this study, we challenged SEA to MAIDS mice and examined whether this retrovirus affects the fate of the SEA-reactive CD4+ T cells in vivo . At 4 wk post-infection with LP-BM5 murine leukemia virus, clonal expression and subsequent deletion of SEA-reactive CD4+V beta 3+ T cells occurred normally after SEA administration, whereas in vitro proliferative responses were severely impaired . At 8 wk postinfection, the in vivo expansion of CD4+V beta 3+ T cells was evident, but not followed by clonal deletion, as late as 14 days after SEA administration . This expanding subset in the infected mice expressed the Fas Ag in the same amount as the same subset in uninfected controls . These findings suggest that activation-induced apoptosis of superantigen-reactive CD4+ T cells is interfered with in vivo during the course of MAIDS, which is not attributable to underexpression of the Fas Ag by the CD4+ T cells.

J Mol Biol, 1994 Oct 14, 243(1), 93 - 9
Residual structure in a staphylococcal nuclease fragment . Is it a molten globule and is its unfolding a first-order phase transition?
Griko YV, Gittis A, Lattman EE, Privalov PL.
Temperature-induced unfolding of staphylococcal nuclease and its large fragment, which lacks 13 C-terminal amino acid residues, was studied calorimetrically, and by CD and fluorescence spectroscopy . It was shown that, in contrast to the full length protein which includes two domains and unfolds in two distinct stages under some conditions, the fragment unfolds in one stage . Unfolding of the fragment proceeds in the same temperature range in which the N-terminal beta-barrel domain unfolds in the full length staphylococcal nuclease . Therefore, the fragment is initially partly unfolded . It retains a stable N-terminal domain which unfolds co-operatively with significant heat absorption . Unfolding of the fragment can be regarded as a first-order phase transition, but its initial state certainly does not represent a molten globule, as it was believed.

J Mol Biol, 1994 Oct 7, 242(5), 670 - 82
Electrophoretic characterization of the denatured states of staphylococcal nuclease; Creighton TE et al.; The denatured state of staphylococcal (staph) nuclease has been shown to be altered by mutations, which complicates studies of stability of the folded state and may be important for the folding process . The relative hydrodynamic volumes as a function of urea concentration of staph nuclease, and of mutant forms in which the native state was substantially destabilized, have been examined by urea gradient gel electrophoresis . The native conformation is unique in its stabilization by specific ligands, which made it possible to distinguish between intermediate electrophoretic mobilities resulting from a partly populated native conformation and from other partly folded conformations . Four distinct conformational states were observed; two native-like: (1) the native, with and without bound ligands, and (2) an altered native-ligand complex at high urea concentrations; plus two denatured states: (3) a partly folded compact conformation that unfolded co-operatively and was sensitive to mutation of residues in the beta-barrel portion of the protein, and (4) the unfolded state, which appeared from its hydrodynamic volume to be as unfolded as reduced, alkylated ribonuclease A, even at very low urea concentrations, and was not altered by single mutations . The effects of mutations on the denatured state of staph nuclease may be due to the occurrence of the partly folded conformation (3).

Biochemistry, 1994 Oct 4, 33(39), 11745 - 59
Solution structure of the active domain of tissue inhibitor of metalloproteinases-2 . A new member of the OB fold protein family; Williamson RA et al.; Homonuclear two-dimensional and three-dimensional 1H nuclear magnetic resonance spectroscopy has been used to obtain essentially complete sequence-specific assignments for 123 of the 127 amino acid residues present in the truncated form of tissue inhibitor of metalloproteinases-2 (delta TIMP-2), the active N-terminal domain of the protein . Analysis of the through-space nuclear Overhauser effect data obtained for delta TIMP-2 allowed determination of both the secondary structure of the domain and also a low-resolution tertiary structure defining the protein backbone topology . The protein contains a five-stranded antiparallel beta-sheet that is rolled over on itself to form a closed beta-barrel, and two short helices which pack close to one another on the same barrel face . A comparison of the delta TIMP-2 structure with other known protein folds reveals that the beta-barrel topology is homologous to that seen in proteins of the oligosaccharide/oligonucleotide binding (OB) fold family . The common structural features include the number of beta-strands and their arrangement, the beta-barrel shear number, an interstrand hydrogen bond network, the packing of the hydrophobic core, and a conserved beta-bulge . Superpositions of the beta-barrels from delta TIMP-2 and two previously known members of the OB protein fold family (staphylococcal nuclease and Escherichia coli heat-labile enterotoxin) confirmed the similarity in beta-barrel topology . The three-dimensional structure of delta TIMP-2 has allowed a more detailed interpretation than was previously possible of the functional significance of available protein sequence and site-directed mutagenesis data for the TIMP family . Furthermore, the structure has revealed conserved surface regions of potential functional importance.

J Orthop Trauma, 1994 Oct, 8(5), 390 - 6
The efficacy of various irrigation solutions in removing slime-producing Staphylococcus; Anglen JO et al.; To determine which type of irrigation solution and which method of irrigation most effectively removes slime-producing Staphylococcus from implant surfaces, we performed experimental washings of bacteria-coated stainless steel screws with various solutions delivered by bulb syringe or by jet lavage . The quantity of bacteria remaining on the screw surface was determined after irrigation with 1 L of saline, 1 L of antibiotic solutions, or 1 L of saline containing a liquid soap . Antibiotic solutions tested included bacitracin, neomycin, and polymyxin/neomycin . We found that the use of power irrigation increased the removal of bacteria by a factor of at least 100 over bulb syringe irrigation of the same volume, no matter which solution was used . This effect ranged from a 100-fold improvement for neomycin, to a 285-fold effect for the polymyxin solution . The addition of antibiotic drugs to the irrigation solution had no significant effect on bacterial removal, and none of the antibiotic solutions were statistically different from saline alone in the amount of bacteria removed from the screws . The addition of a liquid soap solution dramatically increased the amount of bacteria removed by irrigation, reducing the residual bacteria per screw from a colony count of 3.5 x 10(4) for polymyxin (the best of the antibiotic solutions), to 4.38 x 10(3) . This difference was statistically significant as judged by Student's t test, with p = 0.01 . We have concluded that the use of power irrigation improves the ability to clean this pathogenic bacteria from metallic surfaces, and that the addition of antibiotic drugs to the irrigation solution does not.(ABSTRACT TRUNCATED AT 250 WORDS)

J Cardiovasc Surg (Torino), 1994 Oct, 35(5), 395 - 8
Homograft aortic root replacement for destructive valve endocarditis . The benefit of an in-house homograft bank; Fischlein T et al.; This report concerns the surgical treatment of a 45-year old patient with active staphylococcus-endocarditis of the aortic valve which resulted in an aortic root abscess and consequently in a ventricular aortic discontinuity . A technique of homograft aortic root replacement after removal of all infected and necrotic areas is described . After a six month follow-up, the patient is asymptomatic (New York Heart Association functional class I) and shows no signs of recurrence of endocarditis . This case report makes the benefit of an in-house homograft-bank system obvious.

Can Vet J, 1994 Oct, 35(10), 617 - 21
Efficacy of tylosin tablets for the treatment of pyoderma due to Staphylococcus intermedius infection in dogs; Scott DW et al.; Tylosin tablets (20 mg/kg, q12h) were administered orally to 21 dogs with superficial or deep staphylococcal pyodermas . Response to therapy was excellent in 90.5% of the dogs, and in vitro susceptibility testing correlated perfectly with therapeutic response . Duration of therapy varied from 17 to 91 days, with an average of 33 days . Relapses occurred in 28.6% of the dogs within a three-month period . No side effects were reported . Under the conditions of the study, tylosin was an effective and safe antibiotic for the treatment of staphylococcal pyoderma in dogs.

Rofo, 1994 Oct, 161(4), 292 - 9 E
{Radiologic differential diagnosis of inflammatory round pulmonary infiltrates in immunocompromised patients . A prospective study using CT and MRT}; Blum U et al.; In a prospective study we examined the diagnostic ranking of CT and MR in 52 immunocompromised patients with nodular pulmonary lesions and clinical suspicion of invasive pulmonary aspergillosis (IPA) . For early diagnosis of IPA (clinical symptoms having existed for less than 10 days) the CT halo sign proved highly sensitive and specific . MRT showed at this time a comparatively high sensitivity but only low specificity that could not be improved upon after Gd-DTPA . At a later stage of the aspergillosis infection (clinical symptoms manifested for more than 10 days) MR identified aspergillus-specific lesions with on-target characteristics (marked enhancement of margins after Gd-DTPA) or the so-called "reverse" target phenomenon (T2-weighted sequences) . Such lesions were never seen in the early stage of the disease in patients with nodular pulmonary lesions of different aetiology (pseudomonal or staphylococcal pneumonia).

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