Correlation of serum protein biomarkers with disease activity in psoriatic arthritis.

Objective: To assess the correlation of serum protein biomarkers with disease activity across different domains of psoriatic arthritis (PsA).Material and methods: A cross-sectional cohort of 45 adult patients with PsA fulfilling the classification for psoriatic arthritis (CASPAR) criteria was recruited from University of California San Diego (UCSD) Arthritis Clinics. Clinical data and serum samples were collected and serum was analyzed for protein biomarkers hypothesized to be relevant to disease activity in PsA. Correlations were evaluated for clinical disease activity measures across disease domains.Results: Biomarkers with the highest correlation to the composite indices and disease domains were SAA, IL-6, YKL-40, and ICAM-1. In addition, several biomarkers were moderately correlated with individual composite indices and/or disease domains. Low or no correlation was observed with some biomarkers, e.g. MMP-3, MMP-1, EGF, VEGF, and IL-6R. In contrast, the correlation of all biomarkers with certain disease domains was low; specifically, pain, percent body surface area of psoriasis, and patient global assessment. The multi-biomarker disease activity score (MBDA) developed for rheumatoid arthritis (RA) showed high correlations with most composite indices and some disease domains in PsA.Conclusions: These data suggest biomarker analysis can reflect disease activity across disease domains in PsA. Certain domains would likely benefit from the evaluation of additional biomarkers.

Prevalence of age-related macular degeneration in an elderly UK Caucasian population-The Bridlington Eye Assessment Project: a cross-sectional study.

ImportanceThere is paucity of data on prevalence and disease asymmetry of age-related macular degeneration (AMD), particularly the earlier stages, in the UK population.Objective and PurposeTo determine the prevalence of age-related macular degeneration in an elderly Caucasian UK population.DesignCross-sectional population study, 2002-2006.ParticipantsResidents in the study area of Bridlington aged 65 years and older.MethodsFull-ophthalmic examination was undertaken in 3549 participants, of eligible 6319 Caucasian population (response rate of 56%). Non-stereoscopic Colour fundus photographs (30°) were graded masked using a modified Rotterdam Classification for 3475 (98%) participants with gradable images. Prevalence for different AMD grades were calculated. Demographic details were analysed then integrated with the AMD gradings for full analysis. Prevalence rates for the different AMD Grades were calculated, as well as the age-specific prevalences.ResultsAMD prevalence in the worst eye were 38.5% grade 0, 41.4% grade 1, 12.8% grade 2, 2.8% grade 3, and 4.6% grade 4. Geographic atrophy (grade 4a) occurred in 2.5%, and neovascular AMD (grade 4b) in 1.8%. Prevalence increased with age such that grade 4 (advanced) AMD was 2.2% in the 65-69 years group, 15.8% for the 85-90, and 21.2% for over 90 years. There was significant asymmetry between the two eyes of individuals with advanced AMD (P<0.001), such that vision loss was unilateral. Persons with more advanced AMD grades were more likely to be dissatisfied with their vision.ConclusionsAdvanced AMD occurs more commonly in the UK Caucasian population than previously reported. Significant asymmetry between the two eyes occurs in individuals with unilateral advanced AMD so that visual impairment statistics do not represent true prevalence of advanced AMD. Persons with more advanced AMD were more likely to be dissatisfied with their vision.

Peripherally administered oxytocin modulates latent inhibition in a manner consistent with antipsychotic drugs.

BACKGROUND: Peripherally administered oxytocin (OT) has produced antipsychotic drug (APD)-like effects in animal tests that are predictive of APD efficacy. However, these effects have mainly been demonstrated using animal models of schizophrenia-like deficits in prepulse inhibition (PPI) of the startle reflex. Another schizophrenia-relevant abnormality that is the basis of a predictive animal test for APD efficacy is deficient latent inhibition (LI). LI is the normal suppression of a classically conditioned response when the subject is pre-exposed to the conditioned stimulus (CS) before it is paired with the unconditioned stimulus (UCS). Conditioned taste aversion (CTA), the normal avoidance of ingesting a food or liquid by animals when its taste is associated with an aversive experience, was used to test whether OT facilitates LI consistent with APDs. METHODS: Brown Norway rats, known to naturally display attenuated LI, were aversively conditioned on two consecutive exposures to flavored drinking water (0.1% saccharin) by pairing it with malaise-inducing lithium chloride injections. Concurrent with conditioning, rats received subcutaneous OT (0.02, 0.1, 0.5mg/kg) or saline. Some rats were pre-exposed to the flavored water prior to its aversive conditioning (pre-exposed) while others were not (non pre-exposed). Two days after aversive conditioning the amount of flavored water consumed during a 20-min session was recorded. RESULTS: As expected, LI, defined as greater consumption by pre-exposed vs. non pre-exposed rats was only weakly exhibited in Brown Norway rats and OT enhanced LI by reducing CTA in pre-exposed rats in a dose-dependent manner, with the 0.02 mg/kg dose producing the strongest effect. CONCLUSIONS: The facilitation of LI by OT is consistent with the effects produced by APDs and provides further support for the notion that OT has therapeutic potential for schizophrenia.

Validated gene expression biomarker analysis for biopsy-based clinical trials in ulcerative colitis.

BACKGROUND: Accurate and reproducible measurement of expression of pro-inflammatory cytokines in colonic biopsies from patients with ulcerative colitis (UC) is essential for proof-of-concept and mechanism-of-action studies. Few studies have rigorously established the number of biopsies required for accurate and reproducible biomarker measurements. AIM: To validate methods for measuring changes in gene expression in colonic biopsy samples. METHODS: Twelve colonic biopsies were obtained from each of six healthy controls, six patients with inactive UC and seven patients with active UC. Mayo endoscopic scores were used as a clinical reference standard. Quantitative PCR was used to assess mRNA expression of eight known inflammatory genes. The power to detect a reduction in gene expression in active vs. inactive UC was calculated using a linear mixed effect model. RESULTS: mRNA analysis of colonic biopsies is a sensitive and feasible approach for measuring inflammatory gene expression in colonic biopsies. Inflammatory biomarkers correlate with Mayo endoscopic subscores for each colonic region. For most genes, three rectal biopsies from two to four patients are required to detect changes in gene expression corresponding to active vs. inactive UC to achieve a power of 80% with an alpha of 0.05. CONCLUSION: Our data suggest that systematic measurement of inflammatory biomarkers at the mRNA level can be a valuable tool for hypothesis testing, and assessment of clinical activity and response to therapy in ulcerative colitis.

Community optometrist referral of those aged 65 and over for raised IOP post-NICE: AOP guidance versus joint college guidance--an epidemiological model using BEAP.

BACKGROUND/AIMS: To identify the percentage of those aged 65 and over who might be referred by community optometrists as ocular hypertensive suspects in the post-NICE era when differing guidance is followed by community optometrists. METHOD: The authors constructed an epidemiologically based model utilising Bridlington Eye Assessment Project (BEAP) data. Ocular hypertensive suspects' data were subjected to two algorithms (Association of Optometrists (AOP) and Joint College) to determine referral of suspects if community optometrists followed either algorithm. RESULTS: 85 of 1643 people (5.2%) tested by BEAP, with normal acuity and visual fields, recorded Goldmann IOPs of >21 mm Hg in either or both eyes. Without pachymetric information, all 85 would be referred under the AOP algorithm, decreasing to 31 (1.9%) under the joint College algorithm (63% reduction). If central corneal thickness readings influenced referral, 39 (2.4%) would be referred under the AOP algorithm and 13 (0.8%) under the joint College algorithm. CONCLUSION: If community optometrists use Goldmann tonometry and pachymetry, following the joint College guidelines, referrals of OHT suspects could be reduced to a fifth of those under the original AOP guidance. Community optometrists should be encouraged to use GAT and pachymetry in order to refine referrals when another examination is normal. Potential savings to the NHS are considerable.

Acid-sensing ion channel 3 expressed in type B synoviocytes and chondrocytes modulates hyaluronan expression and release.

BACKGROUND: Rheumatoid arthritis is an inflammatory disease marked by intra-articular decreases in pH, aberrant hyaluronan regulation and destruction of bone and cartilage. Acid-sensing ion channels (ASICs) are the primary acid sensors in the nervous system, particularly in sensory neurons and are important in nociception. ASIC3 was recently discovered in synoviocytes, non-neuronal joint cells critical to the inflammatory process. OBJECTIVES: To investigate the role of ASIC3 in joint tissue, specifically the relationship between ASIC3 and hyaluronan and the response to decreased pH. METHODS: Histochemical methods were used to compare morphology, hyaluronan expression and ASIC3 expression in ASIC3+/+ and ASIC3-/- mouse knee joints. Isolated fibroblast-like synoviocytes (FLS) were used to examine hyaluronan release and intracellular calcium in response to decreases in pH. RESULTS: In tissue sections from ASIC3+/+ mice, ASIC3 localised to articular cartilage, growth plate, meniscus and type B synoviocytes. In cultured FLS, ASIC3 mRNA and protein was also expressed. In FLS cultures, pH 5.5 increased hyaluronan release in ASIC3+/+ FLS, but not ASIC3-/- FLS. In FLS from ASIC3+/+ mice, approximately 50% of cells (25/53) increased intracellular calcium while only 24% (14/59) showed an increase in ASIC3-/- FLS. Of the cells that responded to pH 5.5, there was significantly less intracellular calcium increases in ASIC3-/- FLS compared to ASIC3+/+ FLS. CONCLUSION: ASIC3 may serve as a pH sensor in synoviocytes and be important for modulation of expression of hyaluronan within joint tissue.

Preliminary assessment of safety and effectiveness in humans of ProBiora3, a probiotic mouthwash.

AIMS: To conduct a pilot human clinical trial to assess the safety and to test the ability of a probiotic mouthwash, ProBiora(3), to affect the levels of Streptococcus mutans and certain known periodontal pathogens in the mouth when administered twice daily over a period of 4 weeks. METHODS AND RESULTS: The mouthwash contained three specific strains of naturally occurring oral bacteria and was tested at two dose levels: 10(6) and 10(8) colony forming units each of Strep. oralis strain KJ3sm, Strep. uberis strain KJ2sm, and the spontaneous lactic acid-deficient variant of Strep. rattus, strain JH145. Substantial decreases in the levels of the marker bacteria were observed. No safety issues were noted with the twice daily application of this mouthwash. CONCLUSIONS: Despite the small number of subjects and the use of young, orally healthy adults, along with the inherent variability in the microbiological measurements, the probiotic mouthwash was able to substantially affect the levels of dental pathogens in saliva and periodontal pathogens in subgingival plaque. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this pilot human study suggest that the probiotic mouthwash product may be safe for daily use as an aid in maintaining both dental and periodontal health.

A spontaneous lactate dehydrogenase deficient mutant of Streptococcus rattus for use as a probiotic in the prevention of dental caries.

AIMS: To study the ability of daily applications of Streptococcus rattus strain JH145 to affect the numbers of an implanted Streptococcus mutans strain in a rat model. METHODS AND RESULTS: A spontaneous L(+)-lactate dehydrogenase (LDH)-deficient mutant of Streptococcus rattus, JH146, was isolated by screening on selective medium and compared with a previously isolated spontaneous LDH deficient strain, JH145. Both strains were shown to have single base pair deletion mutations in the structural gene (ldh) for LDH, and reversion frequencies were approximately the same. Animals treated once daily with >or=10(6) CFU (colony forming units) of JH145 showed a statistically significant decrease in the proportion of implanted S. mutans to total cultivable bacteria in oral swab samples. The rate of decrease in S. mutans levels was dose-dependent. No adverse effects were observed by in-life observation of treated animals, and histopathological, haematological and blood chemistry analyses were unremarkable. CONCLUSIONS: The results presented indicate that daily application of JH145, a naturally occurring LDH-deficient variant of S. rattus, can compete with S. mutans for its habitat on the tooth surface. SIGNIFICANCE AND IMPACT OF THE STUDY: S. rattus JH145 has potential as a probiotic for use in the prevention of dental caries.

The relationship between central corneal thickness and the optic disc in an elderly population: the Bridlington Eye Assessment Project.

PURPOSE: To determine population-based normative CCT data for elderly white subjects, with and without diabetes, and to explore the relationship between CCT and IOP and HRT2 measurements in such subjects with normal visual fields. METHODS: All eligible subjects were consecutive in a population screened for eye disease. CCT was measured by ultrasound pachymetry, and the optic disc was morphometrically defined using Heidelberg retinal tomography (HRT2). Inclusion criteria were: (1) normal visual field on suprathreshold testing and (2) corrected logMAR acuity of at least 0.3 in both eyes. Subjects with significant corneal pathology, previous corneal surgery, or known history of glaucoma or treatment for raised intraocular pressure were excluded. One eye was randomly selected from each subject for analysis. RESULTS: In all, 983 eyes of 983 subjects were included with 690 HRT images deemed acceptable for the analysis. The mean age (414 men and 569 women) was 73.3 years (minimum 65 years). Mean (SD) CCT was 544.1 (36.5) microm, with a normally distributed range of 429-633 microm. There was no significant difference in CCT between men and women (mean CCT 546.1 and 542.7 microm, respectively, P=0.15, Student's t-test), though CCT was correlated weakly and negatively with age (Pearson's r=-0.063, P=0.047). Diabetic patients (n=103) had a greater mean CCT than non-diabetic patients (551.9 and 543.0 microm respectively, P=0.02). No significant correlation was found between any global optic disc parameter and CCT in the 690 eyes analysed. CONCLUSION: Elderly white eyes with normal fields have CCTs that are normally distributed, with those from diabetic persons having greater CCTs. No clear evidence of a relationship between CCT and HRT2 optic disc parameters used in glaucoma diagnosis was found.

Elucidation of the antimicrobial mechanism of mutacin 1140.

Mutacin 1140 and nisin A are peptide antibiotics that belong to the lantibiotic family. N-Terminal rings A and B of nisin A and mutacin 1140 (lipid II-binding domain) share many structural and sequence similarities. Nisin A binds lipid II and thus disrupts cell wall synthesis and also forms transmembrane pores. Very little is known about mutacin 1140 in this regard. We performed fluorescence-based studies using a bacteria-mimetic membrane system. The results indicated that lipid II monomers are arranged differently in the mutacin 1140 complex than in the nisin A complex. These differences in complex formation may be attributed to the fact that nisin A uses lipid II to form a distinct pore complex, while mutacin 1140 does not form pores in this membrane system. Further experiments demonstrated that the mutacin 1140-lipid II and nisin A-lipid II complexes are very stable and capable of withstanding competition from each other. Transmembrane electrical potential experiments using a Streptococcus rattus strain, which is sensitive to mutacin 1140, demonstrated that mutacin 1140 does not form pores in this strain even at a concentration 8 times higher than the minimum inhibitory concentration (MIC). Circular complexes of mutacin 1140 and nisin A were observed by electron microscopy, providing direct evidence for a lateral assembly mechanism for these antibiotics. Mutacin 1140 did exhibit a membrane disruptive function in another commonly used artificial bacterial membrane system, and its disruptive activity was enhanced by increasing amounts of anionic phospholipids.

Potential use of quantitative bedside CBF monitoring (Xe-CT) for decision making in neurosurgical intensive care.

During a 3-year period, mobile xenon-computerized tomography (Xe-CT) for bedside quantitative assessment of cerebral blood flow was used as an integrated tool for decision making during the care of complicated patients in our neurosurgical intensive care units (NSICU), in an attempt to make a preliminary evaluation regarding the usefulness of this method in routine work in the neurosurgical intensive care. With approximately 200 studies involving 75 patients, we identified six different categories where the use of bedside Xe-CT significantly influenced (or, with more experience, could have influenced) the decision making, or facilitated the handling of patients. These categories included identification of problems not apparent from other types of monitoring, avoidance of adverse effects from treatment, titration of standard treatments, evaluation of the vascular resistance reserve, assessment of adequate perfusion pressure and better utilization of resources from access to the bedside cerebral blood flow (CBF) technology. We conclude that quantitative bedside measurements of CBF could be an important addition to the diagnostic and monitoring arsenal of NSICU-tools.

Modification of an effector strain for replacement therapy of dental caries to enable clinical safety trials.

AIMS: To construct a genetically modified strain of Streptococcus mutans for dental caries prevention. The strain has significantly reduced cariogenicity owing to a deletion of the entire open reading frame for lactate dehydrogenase, and has excellent colonization potential through the production of a natural antibiotic called mutacin 1140. For use in human clinical trials, additional mutations were introduced to enable rapid elimination of the strain in case of adverse side effects and to increase genetic stability. METHODS: Deletion mutations were introduced into the dal gene for d-alanine biosynthesis and the comE gene for genetic transformation. The resulting strain, A2JM, was tested for dependence on exogenous d-alanine and its ability to be eradicated from colonized rats. The strain was also tested for its ability to exchange DNA with another strain of S. mutans in in vitro and in vivo models. CONCLUSIONS: A2JM was completely dependent on exogenous d-alanine, but could colonize the oral cavity of rats in low numbers in the absence of dietary d-alanine. Results indicated that A2JM can scavenge d-alanine from other plaque bacteria. Lowering of the total oral bacterial load through daily application of chlorhexidine enabled virtually complete eradication of A2JM. The introduction of the comE gene did not significantly decrease the transformability of A2JM in in vitro or in vivo models. The addition of a deletion in the comE gene does, nonetheless, provide additional safety as it has a very low reversion frequency. SIGNIFICANCE AND IMPACT OF THE STUDY: Based on the safety and efficacy profiles established in vitro and in animal models, A2JM appears suitable for safe use in human clinical trials.

An alternative bactericidal mechanism of action for lantibiotic peptides that target lipid II.

Lantibiotics are polycyclic peptides containing unusual amino acids, which have binding specificity for bacterial cells, targeting the bacterial cell wall component lipid II to form pores and thereby lyse the cells. Yet several members of these lipid II-targeted lantibiotics are too short to be able to span the lipid bilayer and cannot form pores, but somehow they maintain their antibacterial efficacy. We describe an alternative mechanism by which members of the lantibiotic family kill Gram-positive bacteria by removing lipid II from the cell division site (or septum) and thus block cell wall synthesis.

Intracerebral microdialysis in neurosurgical intensive care patients utilising catheters with different molecular cut-off (20 and 100 kD).

OBJECTIVE: To compare the properties of a new intracerebral micro-dialysis catheter with a high cut-off membrane (molecular cut-off 100 kDalton) with a standard catheter (CMA70, molecular cut-off 20 kDalton). METHODS: Paired intracerebral microdialysis catheters were inserted in fifteen comatose patients treated in a neurosurgical intensive care unit following subarachnoid haemorrhage or traumatic brain injury. The high-cut-off catheter (D(100)) differed from the CMA 70 catheter by the length (20 mm) and cut-off properties of the catheter membranes (100 kDalton) and the perfusion fluids used (Ringer-Dextran 60). Samples were collected every 4-6 hours, analyzed bedside (for glucose, glutamate, glycerol, lactate, pyruvate and urea) and later in the laboratory (for total protein). RESULTS: Fluid recovery was similar for the two types of catheters, but significantly more protein was recovered by the D(100) catheter. The recovery of glycerol and pyruvate did not differ, while minor differences in recovery of glutamate and glucose were observed. The recovery of lactate was considerably lower in the D(100) catheter (p < 0.01), influencing the lactate/pyruvate-ratio. The patterns of concentration changes over time were consistent for all metabolites, and independent of type of catheter. CONCLUSION: Microdialysis catheters with high cut-off membranes can be used in routine clinical practice in the NSICU, adding the possibility of macro-molecule sampling from the extracellular space during monitoring.

Bedside monitoring of CBF with xenon-CT and a mobile scanner: a novel method in neurointensive care.

Combining previously independently established techniques our objective was to develop and evaluate a method for bedside qualitative assessment of cerebral blood flow in neurointensive care (NICU) patients. The CT-protocol was optimized using phantoms and comparing a mobile CT-scanner (Tomoscan-M, Philips) with two stationary CT scanners. Thirty-two per cent xenon was delivered with standard equipment (Enhancer 3000). Mean cortical flow in volunteers was 48 ml/min/100 g, with the mean vascular territorial flow varying between 45 and 66 ml/min/100 g. The potential clinical usefulness was illustrated in three patients with vasospasm following subarachnoid haemorrhage. Our conclusion is that quantitative bedside measurements of CBF can be repeatedly performed in an easy and safe way in a standard NICU-setting, using xenon-inhalation and a mobile CT-scanner. The method is useful for the decision-making, and is a good example of how the quality of multi-modality monitoring in the NICU can be developed and further diversified.

Effect of season of application on the adhesion, retention and recontamination of herbage by potentially toxic metals and sewage sludge.

Grassland in the United Kingdom can accept sewage sludge when, as a result of cropping restrictions, arable land cannot. However, one of the uncertainties associated with the application of sewage sludge to grassland is the potential exposure of the grazing ruminant to sewage sludge particulates adhered to the grass or deposited on the soil surface. The implications of elevated levels of sludge particulates and associated potentially toxic metals (PTMs) are the potential accumulation of PTMs into edible tissues of the grazing animal. Two experiments were performed to investigate the effect of season of application on the adhesion, retention and recontamination of herbage by sewage sludge and PTMs. In each experiment, three treatments were installed, an untreated control, application of sludge in the autumn and application of sludge in the spring. Sewage sludge particulates and PTMs were eluted sequentially 21 days after the application of sludge using water and detergent and the residual (tissue) concentrations were monitored. Total concentrations in each fraction and adhesion indices were calculated. Soil samples taken from the sward treatments after each application showed no elevated concentrations of PTMs or concentrations that approached the maximum concentrations allowed for soils amended with sewage sludge. The amounts of sludge adhered to herbage ranged from 114 to 360 g sludge DM/kg herbage DM. There was evidence of preferential retention of PTMs to herbage. However, during the 21-day no grazing period there was a general decline in concentration of all PTMs. Copper was retained on the herbage for greater periods of time compared to Zn, CD, Pb and Fe (P < 0.01). The proportion of Pb recovered from the water phase increased during the 21-day no grazing period in comparison to Cu, Zn and Fe (P < 0.001). It is concluded that the concentrations of PTMs adhered and integral to herbage as a result of surface application of sewage sludge are likely to fall to acceptable levels in well-managed permanent grassland.

Postoperative pseudoaneurysm of the superficial temporal artery (S.T.A.) treated with Thrombostat (thrombin glue) injection.

BACKGROUND: Pseudo-aneurysm is a rare complication of craniotomy. Blunt injury to the temporal artery region is the usual cause, but still a rare complication. CLINICAL PRESENTATION: A patient with subarachnoid hemorrhage was successfully treated by aneurysm clipping. The patient developed hydrocephalus, and was admitted for a shunt operation seventeen days later. The craniotomy had healed normally, but a palpable temporal lump was present in the skin incision. INTERVENTION: The pulsating mass proved to be a postoperative aneurysm of the superficial temporal artery (S.T.A.) and was successfully occluded with 500 units Thrombostat (thrombin glue) which was injected into the aneurysm sac using a 22-gauge needle guided by ultrasound. The permanency of the obliteration was verified by ultrasound examination.

Moderate hypothermia for 359 operations to clip cerebral aneurysms.

BACKGROUND: Experimental data have suggested that hypothermia (32-34 degrees C) may improve outcome after cerebral ischaemia, but its efficacy has not yet been established conclusively in humans. In this study we examined the feasibility and safety of deliberate moderate perioperative hypothermia during operations for subarachnoid aneurysms. METHODS: A total of 359 operations for intracranial cerebral aneurysms were included in this prospective study. By using cold intravenous infusions (4 degrees C) and convective cooling our aim was to reduce the patient's core temperature to more than 34 degrees C within 1 h before operation. The protocol assessed postoperative complications such as infections, prolonged mechanical ventilation, pulmonary complications and coagulopathies. RESULTS: During surgery, the body temperature was reduced to a mean of 32.5 (SD 0.4) degrees C. Cooling was accomplished at a rate of 4.0 (SD 0.4) degrees C h(-1). All patients were normothermic at 5 (sd 2) h postoperatively. Peri/postoperative complications included circulatory instability (n=36, 10%), arrhythmias (n=17, 5%) coagulation abnormalities and blood transfusion (n=169, 47%), infections (n=29, 8%) and pulmonary complications (infiltrate or oedema while on ventilatory support) (n=97, 27%). Eighteen patients died within 30 days (5%). There was no significant correlation between the extent of hypothermia and any of the complications. However, there was a strong correlation between the occurrence of complications and the severity of the underlying neurological disease as assessed by the Hunt and Hess score. CONCLUSION: Moderate hypothermia accomplished within 1 h of induction of anaesthesia and maintained during surgery for subarachnoid aneurysms appears to be a safe method as far as the risks of peri/postoperative complications such as circulatory instability, coagulation abnormalities and infections are concerned.

Structure and dynamics of the lantibiotic mutacin 1140.

Mutacin 1140 is a member of a family of ribosomally synthesized peptide bacteriocins called lantibiotics (lanthionine-containing antibiotics) and is produced by the Gram-positive bacterium Streptococcus mutans. Mutacin 1140 has been shown to be effective against a broad array of Gram-positive bacteria. Chromatography and mass spectroscopy data suggested that mutacin 1140 forms a small compact structure. Nuclear magnetic resonance (NMR) data and restrained molecular dynamics simulations showed that mutacin 1140 interconverts between multiple structures. Calculations of scalar (J) coupling constants showed the best agreement with experimental values when the entire population-weighted ensemble of structures was used, providing independent support for the ensemble. Representative structures from each major group in the ensemble had a common feature in which they are all kinked around the hinge region forming a horseshoe-like shape, and the regions of flexibility of the molecule were limited and well-defined. The structures determined in this study provide a starting point for modeling the mutacin 1140-membrane interactions and pore formation.

The accumulation of potentially toxic elements in edible body tissues of lambs grazing after a single application of sewage sludge.

The application of sewage sludge to grassland can lead to the accumulation of potentially toxic elements (PTEs) at the soil surface that may be ingested, together with soil and herbage, by grazing ruminants. To investigate the extent to which the method of application of sludge to grassland might influence the accumulation of PTEs in body tissues of grazing sheep, two experiments were undertaken using grassland overlying two contrasting types of soil. At both sites, plots were either untreated or given liquid digested sludge in a single application in either autumn or spring by surface spreading or by injection. Weaned lambs were grazed on the experimental sites for 149 days (Experiment 1) or 160 days (Experiment 2). Muscle, kidney and liver tissues were assayed for concentrations of PTEs at each slaughter date. Applications of sludge were reflected in elevated concentrations of Pb and Cu in soil in Experiment 1 only (P < 0.05). Total loadings of PTEs for the treated grasslands did not exceed UK statutory limits to annual additions to soil. No significant differences were observed in the concentrations of PTEs in herbage between untreated and treated sites, with the exception of the concentration of Cu in herbage after surface application in the spring (Experiment 2). At the end of Experiment 1 the concentration of Cd tended to be higher in kidney tissues of lambs grazed on the pastures amended with sludge in the autumn compared to lambs grazed on untreated pastures. No other treatment effects were evident. Concentrations of PTEs in edible tissues of lambs grazed on grassland given a single dose of sludge were generally low and unlikely to pose a significant risk to the food chain. However, reduced food intake was observed in lambs grazed on grassland which had received sludge recently, indicating that the 21-day UK statutory no-grazing period should be reviewed in relation to grassland given sewage sludge in the spring.