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We have investigated the capacity of N- and C-terminally truncated and chimeric human (h) IgE-derived peptides to inhibit the binding of 125I-labeled hIgE, and to engage cell lines expressing high and low affinity receptors (Fc-epsilon-RI/II). The peptide sequence Pro343-Ser353 of the hC-epsilon-3 domain is common to all h-epsilon-chain peptides that recognize hFc-epsilon-RI. This region in IgE is homologous to the A loop in C-gamma-2 that engages the rat neonatal IgG receptor. Optimum Fc-epsilon-RI occupancy by hIgE occurs at pH 6.4, with a second peak at 7.4. N- or C-terminal truncation has little effect on the association rate of the ligands with this receptor. Dissociation markedly increases following C-terminal deletion, and hFc-epsilon-RI occupancy at pH 6.4 is diminished. His residue(s) in the C-terminal region of the epsilon-chain may thus contribute to the high affinity of interaction. Grafting the homologus rat epsilon-chain sequence into hIgE maintains hFc-epsilon-RI interaction without conferring binding to rat Fc-epsilon-RI. hFc-epsilon-RII interaction is lost, suggesting that these residues also contribute to hFc-epsilon RII binding. h-epsilon-chain peptides comprising only this sequence do not block hIgE/hFc-epsilon-RI interaction or engage the receptor. Therefore, sequences N- or C-terminal to this core peptide provide structures necessary for receptor recognition.

Patients with severe burns are susceptible to infection with Gram-positive organisms including methicillin-resistant Staphylococcus aureus, and often require higher antibiotic dosages compared with other patients. This study examined the pharmacokinetics of a single iv dose of teicoplanin (12 mg/kg) in 15 adults and five children with severe burns. Adults were aged 21-82 years with a median total body surface area (TBSA) burn of 30% (range 15-60%). Children were aged 10 months-l0 years with median TBSA burn of 15% (10-30%). At 12 h, the median serum teicoplanin concentration was 12.8 mg/L (9.027.1 mg/L) in adults and 7.6 mg/L (6.6-l0.8 mg/L) in children, (P < 0.01); at 24 h, the corresponding values were 8.3 mg/L (4.6-l2.9 mg/L) and 5.2 mg/L (4.2-6.0 mg/L). Using a three-compartment model, the median terminal half life in adults was 114 h (47-278 h). Children fitted a two-compartment model with a terminal half-life of 38 h (2l-41 h). The median concentration of teicoplanin in fluid from the burn wound was 60% of the serum antibiotic concentration. A single iv dose of 12 mg/kg of teicoplanin was sufficient to produce therapeutic serum concentrations in burn patients for 24 h, but monitoring of antibiotic levels in serum may be advisable in those with high total clearance, especially children.

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

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

With the decline in incidence of diphtheria in Europe and the USA, many laboratories no longer routinely culture throat swabs for Corynebacterium diphtheriae. However, there is an outbreak of infection with toxigenic strains in Russia and most adults do not have protective levels of antibody. Non-toxigenic strains are known to cause local disease and lysogenic conversion probably occurs in vivo as well as in vitro. Non-toxigenic C. diphtheriae var. gravis, formerly quite rare, has been isolated with increasing frequency in the UK over the last five years. During prospective screening at one Sexually Transmitted Disease Clinic, six (1%) of 578 homosexual men were found to harbour the organism in the throat, four of them with clinical pharyngitis. Only one of 1696 heterosexual men and women were found to be carriers. Seven cases of endocarditis due to this organism were reported in a single year in Sydney, Australia and non-toxigenic C. diphtheriae var. mitis has caused four cases of endocarditis in Switzerland. Non-toxigenic strains are responsible for pharyngitis and occasional invasive disease and should be treated. Routine screening of throat swabs should not be abandoned.

Treatment with an aminoglycoside plus flucloxacillin is commonly used to prevent wound infection and endocarditis after cardiac surgery. Cardiopulmonary bypass, blood transfusion and the lack of a steady state greatly affect handling of gentamicin. Urinary gentamicin excretion is not predictable in terms of preoperative risk factors possibly because there is no clear relationship between serum levels and gentamicin clearance. A study was performed to determine whether the existing prophylactic regimen gave adequate serum levels during surgery and to compare renal excretion of gentamicin and the trough serum levels. Ten patients received gentamicin (1.5 mg/kg at the start of surgery followed by 80 mg tds for 2 days) and flucloxacillin 500 mg qds for 2 days. Serum and urinary concentrations of gentamicin were assayed during surgery and in the early postoperative period. The median apparent serum half-life during the first 8 h was 2.5 h (95%CI 1.7-3.2 h). The median gentamicin clearance was 37 mL/min(95%CI 23-64 mL/min) and the creatinine clearance 85 mL/min (95%CI 72-210 mL/min). Serum levels remained above 1 mg/L during surgery but urinary concentrations varied between 0.4 and 364 mg/L (median 70 mg/L). At 24 h (but not 8 or 16 h), trough serum levels appeared to be related to the amount of gentamicin excreted but the relationship was not quite statistically significant (P = 0.057). Despite the effects of cardiopulmonary bypass, therapeutic serum gentamicin levels were maintained during surgery and reduced renal excretion in the postoperative period was associated with raised levels.

Aminoglycosides are commonly used with flucloxacillin in the prevention of wound infection and endocarditis after cardiac surgery. Earlier studies suggested that the use of aminoglycosides is associated with a small but significant rise in serum creatinine. A regression analysis was performed to identify the preoperative and postoperative factors of importance in determining serum gentamicin concentrations. Serum assays might then be confined to those at particular risk of elevated serum levels. Patients received gentamicin 1.5 mg/kg at the start of surgery followed by 80 mg tds for two days in addition to flucloxacillin. Trough and peak serum gentamicin concentrations were assayed on the first and second days after surgery and urine concentrations were measured with each full catheter bag. Patient characteristics, drug therapy, operation, fluid balance and routine investigations were recorded. A total of 95 of 104 patients were evaluable. Linear discriminant analysis of factors selected by forward stepwise regression identified ten of the 15 patients with subsequent elevated gentamicin concentrations, by using age, and preoperative serum creatinine, systolic blood pressure and serum aspartate transaminase. After operation, factors such as serum creatinine, the quantity of frusemide administered in 24 h, and the integral of the systolic blood pressure over 24 h identified 12 of the 15 patients. However, no satisfactory model could predict urinary concentrations. The discriminant function using preoperative factors prospectively identified 11 of 12 patients with elevated trough levels from a total of 101 further patients. Postoperative gentamicin concentrations cannot reliably be predicted from patient characteristics. Trough assays should be performed or prophylactic courses limited to 24 h.

Activated B cells have recently been shown to produce soluble CD23 from their membranes. The serum-soluble CD23 concentration in 21 patients with pelvic pain diagnosed as having endometriosis and confirmed by histology, and in 18 patients without pelvic pain, who had a normal pelvis during laparoscopic sterilization, was studied by chemiluminescent enzyme-linked immunosorbent assay. The endometriosis patients were randomized to 3 months of either danazol or leuprolide acetate injection. Serum was taken before and after 3 months of therapy. The serum-soluble CD23 concentration was significantly elevated in patients with endometriosis when compared with the controls (P < 0.0001). There was no correlation between soluble CD23 concentrations and the severity of endometriosis (r = 0.48, P > 0.05). The serum concentration of soluble CD23 decreased significantly on treatment with danazol but not leuprolide acetate (P < 0.05). We conclude that the elevation of soluble CD23 in patients with endometriosis suggests that there is activation of B cells, which respond to danazol but not leuprolide acetate injection.

Despite advances in the use of topical and parenteral antimicrobial therapy, and the practice of early tangential excision, infection of the burn wound remains a leading cause of morbidity and mortality. The aim of this study was to survey and compare wound management and antibiotic usage in burn surgery in all UK burn centres. A postal questionnaire was used, followed up by telephone. Answers were obtained from all 39 units treating burns in the UK. A written policy on antibiotic usage was used by 13 centres. Excisions were covered routinely by prophylactic antibiotic therapy in 18 units, and three of these used antibiotic cover during all procedures in which the wound was manipulated. No routine antibiotic cover was given in 21 units. All units obtained surface swab cultures, but only two performed punch biopsy of the wounds. Quantitative bacteriology (counting colony forming units) was employed by three centres. Most units cleaned the burn with saline (17 of 39) or chlorhexidine (eight of 39) but combinations of these and other agents were also used. Nearly half (17 of 37) of those who replied had not seen any cases of proven or suspected toxic shock syndrome in the past 2 years. Four units gave prophylaxis against Staphylococcus aureus, and four gave antibiotics against Streptococcus pyogenes, despite recommendations in the literature. Twenty-nine units gave no prophylaxis. This study has revealed that there is no consensus on antibiotic usage amongst centres treating burns in the UK and that most units rely on surface swab cultures to monitor infection.

Demands are increasing for accurate rates of wound infection in surgical units. However, definitions vary widely and resources are limited. Wound scoring methods improve objectivity and have the power to show changes with time, but they can be laborious. Without community surveillance, inpatient rates can also be misleading.

The validity of the methylene blue colorimetric microassay for determining the response of monolayers of human ovarian tumour cell lines to different growth factors was investigated. Linearity of the relationship between cell density and optical density was confirmed for each cell line (r=0.989-0.999,p<0.001), and when initial cell density was optimised to give exponential growth over the assay period, differences in response to medium supplements were obvious. The response of target cells to growth factors, obtained using the methylene blue assay, were compared with, and found to parallel, previously documented responses obtained non-colorimetrically. Thus Mink lung epithelial cells (MLEC) were inhibited by TGβ (Holleyet al., 1983), EGF had an inhibitory effect on A431 cells (Gill & Lazar, 1981; Barnes, 1982), and the mesothelial cell line showed a proliferative response to EGF and hydrocortisone (Connell and Rheinwald, 1983).The methylene blue colorimetric microssay was found to be a simple, reliable, sensitive method with low variability, for determining the response of cultured cells to growth factors.

This study evaluated a single intradermal injection model in the guinea pig with subsequent inhalation challenge and serological analysis as a method to predict the potential of chemicals to induce respiratory allergy. Four known respiratory allergens (trimellitic anhydride, diphenyl methane diisocyanate, phthalic anhydride and toluene diisocyanate (TDI)) were screened by two industrial research laboratories using this protocol. Dinitrochlorobenzene, a potent contact allergen, was included as a negative control material. In both laboratories, the respiratory allergens, but not the contact allergen, induced high titre antigen-specific antibodies in treated animals. The inhalation challenge results were similar in both laboratories but were less conclusive in that exposure to free TDI failed to induce pulmonary responses, probably because it fails to penetrate to the deep lung in sufficient concentration. Although the assay shows promise as a means of identifying chemical respiratory sensitisers, its use as a routine screen for the prediction of the ability of materials to induce respiratory allergy in man is probably questionable.

The purpose of this study was to investigate the use of the tetrazolium salt. XTT, in a microassay for quantitation of anchorage-independent growth of cells in soft agar, using two human ovarian tumour cell lines (OAW 42 and OAW 28). The response of OAW 42 to Cis-platinum, EGF, TGF β and insulin, and of OAW 28 to EGF, TGF β and insulin were determined both by visual colony counting and use of the XTT assay. Drug inhibition and growth factor inhibition and stimulation were clearly demonstrated, with increases or decreases in visually counted colony numbers being paralleled by increases or decreases in optical density values obtained by using XTT. The XTT colorimetric assay was simple to perform and reproducible with low variability, and was found to be an acceptable alternative to visual colony counting for the quantitation of cell line response to drugs and growth factors under anchorage-independent growth conditions.

The major cause of failure of continuous ambulatory peritoneal dialysis is peritoneal or exit site infection with Staphylococcus aureus. From 1989-1992, eradication of nasal and perineal carriage, continued use of an occlusive dressing to the exit site, improved aseptic technique for dressing changes by the patient and avoidance of wetting of the dressing were used in an attempt to reduce staphylococcal infections. By comparison with the 3-year period prior to intervention, a significant increase in the life of the catheters was achieved (removed at 1 year, 13 vs. 28%, P < 0.001) with a reduction in episodes of peritonitis due to S. aureus. The study was not prospectively controlled but there did not appear to be any other factor to account for the sudden and consistent improvement observed.

Soluble silicates represent a broad range of chemical substances, which, to meet EEC laws, must be classified and labelled to identify their toxicological hazards. The wide range of this type of substance which can be manufactured, and the potentially severe effects which can be produced in standard eye irritation tests, means that classification by standard animal procedures is undesirable. One of the most reliable in vitro assays for assessing eye irritation potential is the rabbit enucleated eye test. This procedure has been used to test a range of soluble sodium silicates to enable these materials to be classified for eye hazard. The experimental approach demonstrates an important example of how an in vitro procedure can be used to propose classification for materials in a reasonable, scientifically valid way, taking account of the nature of the potential hazards and the intrinsic limitations of the in vitro methods used.

Antibiotics are used in 80% of patients in the ICU, encouraging nosocomial infections with resistant organisms. If the antibiotic susceptibilities of the pathogen are known, a narrow-spectrum antibiotic is preferable to preserve the patient's resistance to colonization. However, treatment is often empirical and broad-spectrum combinations are commonly used. Gram-positive bacteraemia is associated with invasive monitoring or intravascular catheters. If the device cannot be removed easily, the glycopeptides are the only agents likely to be active against most strains of the commonest pathogen, the coagulase-negative staphylococcus. Long-stay patients are susceptible to infection with enterococci and methicillin-resistant Staphylococcus aureus, which are often resistant to all the usual agents other than glycopeptides. Vancomycin is long established, but is nephrotoxic, requires serum monitoring, must be administered as an infusion and can cause red man syndrome. Teicoplanin can be given as a single daily bolus without similar side-effects or monitoring. In deep-seated staphylococcal infection, the usual dose of teicoplanin is adequate if given in combination with other agents, but it may need to be doubled if used as monotherapy. Monitoring of the levels in the serum is helpful to ensure an adequate dose in patients with renal failure or in drug abusers, but is not needed to prevent toxicity.

Administration of parenteral antibiotics to outpatients is increasingly used to reduce hospital costs, to reduce loss of earnings for the patient and to improve the quality of life in patients requiring prolonged antibiotic treatment. The glycopeptides are required for treatment of infections caused by methicillin resistant staphylococci and some enterococci, or for treatment of patients allergic to beta-lactam agents. For home therapy, teicoplanin has some advantages over vancomycin in that it requires only once-daily bolus administration, does not necessitate monitoring of serum concentrations and offers the choice of intravenous or intramuscular administration. Teicoplanin has been used to complete treatment of endocarditis at home in selected patients, streptococcal disease being the most suitable form of endocarditis for this treatment. In open trials, teicoplanin has been found effective in home therapy of osteomyelitis but, as with other agents, prolonged dosage can be associated with adverse effects. It has also been used for home treatment of infections of the respiratory tract, intravascular catheters and soft tissue. Despite its higher acquisition costs, teicoplanin is to be preferred over vancomycin because of the reduced administration and assay costs and fewer adverse effects.

The glycopeptide antibiotic, teicoplanin, is increasingly used in Europe in the treatment of Gram-positive infection. It is administered as a bolus once daily, it has little potential for nephrotoxicity, and serum monitoring is usually unnecessary. However, poor results were reported in early trials at a daily dose of 200 mg and, more recently, at 400 mg/day in monotherapy of staphylococcal endocarditis. While 400 mg (6 mg/kg day(-1)) is now standard, US trials have tried very high doses in an attempt to improve its efficacy in monotherapy of deep-seated staphylococcal sepsis. European centres continue to use 6 mg/kg day(-1) as the usual maintenance dose and 6-12 mg/kg as the loading dose. For the more difficult cases, teicoplanin is used in combination with other agents. All available published and unpublished literature was reviewed to try to solve these problems. With the exception of endocarditis, failure rates in the 84 European open studies varied more between trials than between the dosages used. In 32 European and eight US randomized trials, a dose of 6 mg/kg day(-1) of teicoplanin was effective, except in staphylococcal endocarditis if teicoplanin was used as monotherapy. In that case, 12 mg/kg day(-1) or more was needed to achieve a cure rate similar to that of vancomycin. Treatment was most successful with trough levels over 20 mg/l. However, lower doses were effective in combination with aminoglycosides, as is common in clinical practice. An open trial suggested that 12 mg/kg day(-1) was needed for treatment of septic arthritis. It is suggested that 6 mg/kg day(-1) of teicoplanin be used for all indications except staphylococcal endocarditis and septic arthritis when it should be given in a dose of 12 mg/kg day(-1) or in combination with other agents.

A review of the published literature detailing the clinical use of amoxycillin/clavulanic acid spanning the period 1979 to 1992 was undertaken to assess the clinical efficacy of the product and to determine whether any changes had occurred during this time. In the 415 publications meeting the selection criteria a total of over 38,500 patients were treated with amoxycillin/clavulanic acid. Analysis of the data confirms the efficacy of amoxycillin/clavulanic acid over a wide range of clinical indications and annual and triennial groupings of publications suggests that there has been little change in the clinical effectiveness of amoxycillin/clavulanic acid. Clinical efficacy rates (cure or improved) with amoxycillin/clavulanic acid were 88% and 92% in comparative and uncontrolled trials, respectively. Gastro-intestinal side effects are the most common adverse event but have been relatively infrequent. Amoxycillin/clavulanic acid should continue to be a useful antibiotic for upper and lower respiratory tract infections, skin structure infections, dental, head and neck infections, and selected urinary tract infections.

Despite potential nephrotoxicity and ototoxicity, aminoglycosides are widely used in the treatment of severe sepsis but monitoring of serum levels is necessary. The use and assay of aminoglycosides, together with microbiologists' and pharmacists' advice on dosage and potential toxicity were examined in a teaching hospital group during an eight week period. A total of 480 courses of aminoglycoside was recorded in 440 hospital in-patients and of these, 306 (64%) were for prophylaxis. For 79 (45%) of 174 therapeutic courses, bacteriological results were available at the start and the choice of antibiotic was appropriate in 69 cases. Serum aminoglycoside assays were performed in 86% (149/174) of therapeutic courses. Correct assay request forms were used in 79% of 473 assay requests, and the clinical diagnosis was given on 73% of forms. In so-called peak samples with a documented time of collection, this ranged from 5 min to over 9 h post dose. Potentially toxic concentrations were present in 56 (12%) serum samples from 50 courses, in ten of which there was a serum creatinine rise of > 40 microM. However, there were confounding factors in five cases. Probable nephrotoxicity was rare, although assays were not performed in 14% of therapeutic courses. There were no symptomatic cases of ototoxicity but audiometry and vestibular studies were not performed. Closer liaison between the microbiologists, pharmacists and clinical staff is essential to improve clinical practice. Computer notification of inadequately monitored courses would be helpful. The routine therapeutic use of aminoglycosides needs to be reviewed in the light of the rising costs of assay litigation and the increasing number of alternative antibiotics available.

The aim of the study was to determine whether aspirin influences sympathoadrenal output in normal human subjects. Plasma and platelet adrenaline and noradrenaline levels were measured before and after chronic administration of oral aspirin (300 mg per day for 7 days). Catecholamine concentrations measured immediately following aspirin did not differ from control (pre-treatment) values. Platelet noradrenaline and plasma adrenaline levels were, however, significantly increased 2 weeks after cessation of treatment. Platelet TxB2 generation was significantly reduced following aspirin treatment indicating that platelet cyclooxygenase had been inhibited. Catecholamine concentrations did not correlate with TxB2 generation. In vitro platelet aggregation induced by ADP, adrenaline and collagen was reduced after aspirin providing additional confirmation of cyclooxygenase inhibition. However, the in vivo markers of platelet function, beta-TG and PF4 were unaffected. These data do not provide convincing evidence for an action of aspirin on sympathoadrenal outflow, either directly or via a prostaglandin (thromboxane) mediated effect, although this does not exclude a later, delayed effect. There was no evidence for interactions between thromboxane, catecholamine levels in plasma and platelets, and platelet function.

Cell surface expression of the alpha chain of the human (h) Fc epsilon receptor type 1 (Fc epsilon R1) can be observed following stable transfection of RBL-2H3 cells with the h alpha chain of the Fc epsilon R1 complex. Association and dissociation constants for hIgE are similar to those previously reported for the ligand and its receptor. The demonstration of mediator release following the cross-linking of hFc epsilon R1 alpha by hIgE and antigen or anti-hFc epsilon R1 alpha antibody suggests that this system will assist the identification of structural motifs and specific amino acids that are involved in the generation of the signal(s) that will initiate and/or propagate the secretion of mediators from mast cells and basophils.

A throat swab from a 9 year old girl with pharyngitis yielded a non-toxigenic strain of Corynebacterium diphtheriae var mitis and Streptococcus group G. C pseudodiphtheriticum was isolated from the throats of two of her four brothers. In each case the isolate was sent to the reference laboratory before full identification. The growth was found to be mixed for one brother; the other isolate being a toxin producing C diphtheriae var gravis. The child was asymptomatic and the case proves that all colonial types on the Hoyles plate should be identified.

The efficacy of amoxycillin-clavulanic acid as antibiotic prophylaxis in surgery has been assessed in numerous clinical studies, chiefly in abdominal and gynaecological surgery. A meta-analysis of 21 trials covering 2685 patients given amoxycillin-clavulanic acid and 2220 patients given comparator regimens is presented. Monotherapy with amoxycillin-clavulanic acid was as effective as the comparators, including combination regimens utilizing gentamicin or metronidazole, in preventing wound infections (median wound infection rates were 6% and 10% respectively). The antibacterial activity of amoxycillin-clavulanic acid covers the broad range of aerobic Gram-negative and anaerobic organisms that have a major role in postoperative infections. In addition, amoxycillin-clavulanic acid may have benefits in terms of convenience, tolerance and cost.

Significant urinary isolates have been prospectively recorded since 1971. Enterococcus species, a common cause of nosocomial urinary tract infection, have been identified, and susceptibilities to a range of antibiotics have been determined. In addition, isolates in 1988 were tested for breakpoint susceptibility to vancomycin and teicoplanin. Despite changes in the hospitals covered, isolation of Enterococcus species rose steadily from 4% in 1971 to 12.6% in 1990 in hospital patients and from 2% to 5.6% in general-practice patients (P less than .01). All isolates of Enterococcus species were sensitive to ampicillin. Teicoplanin inhibited all 526 strains tested at a concentration of 2 micrograms/mL, but the same concentration of vancomycin inhibited only 370 (70%). The increased prevalence of enterococcal urinary tract infection is probably the result of increasing use of catheterization and broad-spectrum antibiotics. Glycopeptides reach high levels in the urine, and teicoplanin might be an alternative for the treatment of urinary tract infections due to enterococci.

A 41-year-old man who habitually slept in a car park presented with a culture-negative endocarditis. Serological tests indicated infection with both Coxiella burneti and Chlamydia psittaci. He was treated with doxycycline and clindamycin and required aortic valve replacement. Culture of the excised value for both agents was negative but electron microscopy was suggestive of coxiella endocarditis.

Metabolic labeling of the murine T lymphoma cell line RDM-4 with [35S] sulfate results in intense incorporation into a cell-retained component of apparent Mr approximately 100,000. This macromolecule is identified as a glycoprotein by lectin chromatography. The sulfate is not incorporated as tyrosine sulfate. Release of the radiolabel by alkaline beta-elimination but not by endoglycosidase F is consistent with the sulfation of O- rather than N-linked oligosaccharides. The sulfated glycoprotein displays anomalous migration on SDS-PAGE in two respects: 1) the apparent Mr shifts from 115,000 to 87,000 on increasing the acrylamide concentration from 7 to 12%, and 2) on neuraminidase digestion migration is substantially reduced (apparent Mr 140,000). These properties indicate that the sulfated protein is both heavily glycosylated and extensively sialylated, and are characteristic of the lymphoid mucin, leukosialin (sialophorin, CD43). Specific labeling of the sialoglycoproteins of RDM-4 cells indicates that leukosialin, the most intensely labeled protein, comigrates with the sulfated protein on SDS-PAGE at varying acrylamide concentrations. Our data are therefore consistent with sulfation of at least some of the numerous O-linked oligosaccharides of this abundant glycoprotein in RDM-4 cells. No sulfation of CD43 in resting splenic T cells is observed.

Normal murine splenic T lymphocytes and T-lymphoma cells were incubated with [35S]sulphate in low-sulphate medium for 4 hr. Gel filtration and SDS-PAGE revealed that the radiolabelled macromolecules secreted by these cells were almost exclusively chondroitin sulphate and heparan sulphate proteoglycans of relatively low molecular weight (MW), 100,000-200,000. Triton X-100 extracts of the cells contained similar proteoglycans. Under the conditions employed the incorporation of radiolabel by cells grown in vivo was equally distributed between cell-retained and secreted fractions, whereas cells grown in vitro retained some 75% of incorporated label. In general heparan sulphate predominated over chondroitin sulphate in both secreted and cell-retained fractions. Cell extracts also contained a minor proportion of free glycosaminoglycan, which is almost exclusively heparan sulphate. These chains, like those incorporated into the proteoglycan, were around 12,000 MW. The T-lymphoma cells RDM-4, whether grown in vitro or in vivo, also incorporated a substantial proportion of [35S]sulphate into a single, cell-retained protein, 100,000 MW. No such radiolabelled protein was detectable in T cells.

We have reviewed experience with aminoglycosides in cardiac surgical prophylaxis from 1984 to 1989. In the first two years, prospectively randomized trials (517 patients) allowed comparison of tobramycin (three-day course) with a non-toxic antibiotic, teicoplanin. A significant excess rise in serum creatinine was present at the end of the first postoperative week in patients given tobramycin (165 vs 149 patients, 95% CI 3-17 microM. P less than 0.01, which was most marked in ten oliguric patients with trough serum levels over 2 mg/l (95% CI 7-52 microM). However, no patient needed haemodialysis and only one case of possible ototoxicity was identified. During the next three years, of 912 patients given two to three days gentamicin, 29 patients were shown to have potentially toxic serum levels and oliguria. Three individual cases are described, including one of ototoxicity and one of nephrotoxicity neeeding haemodialysis. The hazards of aminoglycoside toxicity are probably small compared with the failure of other antibiotics to cover staphylococci that may later cause wound infection or endocarditis. However, surgeons should probably be advised to check the aminoglycoside level in all patients on the first post-operative day.

For the purposes of wound surveillance programmes and clinical trials, a wound scoring method, ASEPSIS, makes assessment of wound sepsis more objective and reproducible by allotting points both for the appearance of the wound in the first week and for the clinical consequences of infection. ASEPSIS was compared with other definitions of wound infection in 1029 surgical patients and its suitability for surveillance and detection of risk factors were examined. Satisfactory healing was recorded in 867 patients, disturbance of healing in 74 and minor, moderate and severe wound infection in 41, 24 and 23 patients respectively. An ASEPSIS score over 20 points was more sensitive and as specific as the presence of pus as an indicator of changes in management resulting from infection. Multiple regression analysis of ASEPSIS scores indicated that operation type, ward, degree of contamination, age, body mass index, and preoperative stay in hospital were significant risk factors. In matching 52 infected patients with uninfected controls, any wound score over 10 points was associated with a significant delay in discharge from hospital (median 3 days, P less than 0.0005).

The kinetics of binding of oestradiol and the steroidal pure antioestrogen ICI 164,384 to the molybdate-stabilized oestrogen receptor, partially purified from pig and human uterine tissue, were determined. ICI 164,384 bound directly to the oestrogen receptor protein and the kinetic parameters of this interaction were, in general, similar to those for the binding of oestradiol, regardless of the source of the receptor protein. However, the rate of association of oestradiol, regardless of the source of the receptor protein. However, the rate of association of the antagonist with the receptor protein was slower when compared to that of oestradiol. Furthermore, the concentration of binding sites for the two ligands was of the same order. The binding of oestradiol resulted in a steroid-receptor complex which could be transformed in vitro, to a form with increased affinity for DNA-cellulose. However, the complex formed between ICI 164,384 and the receptor protein did not show increased affinity for DNA-cellulose when exposed to conditions that transformed agonist-receptor complexes. Therefore, the binding of ICI 164,384 to the oestrogen receptor protein results in a suppression of the transformation process. A similar suppression in vivo may account for the pure antagonist properties of ICI 164,384.

Cultures of nasal or presternal swabs form part of the routine preoperative screening of patients on the cardiac surgical ward. During a trial of antibiotic prophylaxis in 314 patients, preoperative isolates of Staphylococcus aureus and coagulase-negative staphylococci were compared with strains associated with postoperative sternal wound breakdown (24 patients) and prosthetic valve endocarditis (3 patients). Morphology, antibiotic sensitivity pattern, plasmid analysis and phage typing were used to differentiate strains. In only three cases of wound infection and one of prosthetic valve endocarditis were pathogenic staphylococci not distinguishable from preoperative isolates. The collection of superficial swabs for this purpose before cardiac surgery is therefore unlikely to be cost effective.

A 63-year-old woman with seropositive rheumatoid arthritis was successfully treated with auranofin for 12 months. Heavy proteinuria then developed and the drug was stopped. However, the proteinuria apparently became worse. This was later recognised to be a spurious effect due to the introduction of tolmetin. Renal biopsy showed a type I membranous glomerulonephropathy.

Two prospective trials suggested that teicoplanin was less effective in preventing Gram-positive wound infections after cardiac surgery than a combination of flucloxacillin and an aminoglycoside. The initial dose of 400 mg was associated with a subinhibitory concentration of teicoplanin in fat at the end of operation. In this study, the behaviour of a higher initial dose (12 mg/kg) was investigated in ten patients during coronary artery surgery. As in the earlier trials, a second dose (400 mg) was given after 24 h. The mean concentration at the end of cardiopulmonary bypass was 15 mg/l in serum, 6 mg/kg in fat and 9 mg/kg in skin, all in excess of the expected break-point for staphylococci (4 mg/l). The higher dose regimen might be more successful in surgical prophylaxis.

A controlled trial of antibiotic prophylaxis in cardiac surgery compared a two-dose regimen of teicoplanin with a longer conventional course of flucloxacillin and tobramycin. In 12 patients the susceptibility of the bacterial skin flora of four different sites to each of the three antibiotics was determined and the results are reported here. Less than 1% of the Gram-positive colonies showed reduced sensitivity to teicoplanin (MIC greater than or equal to 4 mg l-1). Before operation, 99% inhibition of Gram-positive growth was achieved at 26 (54%) of 48 sites by 1 mg l-1 of flucloxacillin and 13 (27%) sites by 2 mg l-1 tobramycin. By the 7th day after operation there was a significant reduction in the number of sites showing similar sensitivity to flucloxacillin [16 (33%) sites, P less than 0.05]. The use of teicoplanin was not associated with the emergence of Gram-negative skin flora but tobramycin promoted acquisition of aminoglycoside-resistant strains.