Implementation of the infection control estimate: A case study on the use of a newly developed digital tool for outbreak management in the acute setting.

Aim: An Infection Control Estimate (ICE) Tool was developed based on a previously published concept of applying military planning techniques to Infection Prevention and Control (IPC) management strategies in the acute healthcare setting. Methods: Initial testing of the outbreak management tool was undertaken in a large acute hospital in the North-West of England during a localised outbreak of COVID-19. The tool, developed using Microsoft Excel, was completed by trained IPC practitioners in real-time to log outbreak details, assign and manage meeting actions and to generate surveillance data. Results: The ICE tool was utilised across five outbreak control meetings to identify and allocate tasks to members of the outbreak control team and to monitor progress. Within the meetings, the tool was used primarily by the trained IPC Specialist Nurses who were guided by and entered data into the relevant sections. Feedback indicated that the tool was easy to use and useful as the sole repository of outbreak information and data. Suggested improvements following the testing period were made and additional functionality was added. Conclusion: Utilisation of the ICE tool has the potential to improve our understanding of the efficacy of currently employed outbreak management interventions and provides a cognitive support and targeted education for teams responsible for the management of outbreaks. It is hoped that by guiding teams through an outbreak with prompts and guidance, as well as facilitating collection and presentation of surveillance data, outbreaks will be resolved sooner and risks to patients will be reduced.

Metacognitive training for negative symptoms: Support for the cognitive model.

Developing effective treatment options for negative symptoms of psychotic disorders remains a major unmet treatment need and area for further research. In a recent uncontrolled study by the main author, Metacognition Training (MCT) for negative symptoms was found to lead to fewer negative symptoms, less stigma and increased self-rated reflective ability. As the analysis examined negative symptoms as a whole, we here performed an additional analysis on individual negative symptom items as recent research has suggested that negative symptoms are best conceptualized through a five-factor model. It was found that the intervention led to changes specifically on sociality and blunted affect (with large effect sizes), which might reflect changes in both intrapersonal and interpersonal (meta)cognitive processes.

Fluorescence spectroscopy analysis of fly ash removal from aqueous systems: adsorption of alginate to silica and alumina.

Fly ash is a toxic industrial waste, mainly consisting of silica and alumina particles, that has been found discharged into the environment. It is proposed that alginate, a naturally occurring biopolymer, can bind to these minerals and thus play a role in water purification. The binding forces involved in this process consist of weak interactions, such as van der Waals forces and electrostatic interactions. Although the attachment of alginate to mineral surfaces is mainly governed by its carboxylate groups, hydroxyl moieties could play a role in the interaction between the polymer and minerals. This work aims to use the SiO2 and Al2O3 particles as models for fly ash and to show the use of alginate biopolymers (fluorescently labelled with an aminonaphthaline sulfonate fluorophore (AmNS)) to coagulate them. The addition of simple electrolytes like NaCl and CaCl2 encourages the coiling of the polymer chain at high pH values which has an effect on its capability to bind to the inorganic particles. A combination of fluorescence and ICP-MS demonstrated that alginate has a considerable adsorption affinity for Al2O3, whereas it attracts SiO2 weakly. The adsorption process is pH dependent: strong adsorption was observed at low pH values. The dependence of adsorption on the mineral (Al2O3 and SiO2) concentration was also examined under different pH conditions: the adsorption amount was observed to increase by increasing the solid concentration. Adsorption isotherms obtained at low and high mineral concentrations were found to be Henry in type.

Metacognitive training modified for negative symptoms: A feasibility study.

OBJECTIVE: Although patients often prioritize the treatment of negative symptoms, few psychological interventions targeting negative symptoms exist. This study attempts to fill this gap by piloting a modified metacognitive training programme, specifically targeted at negative symptoms (MCT-N), with a group of patients with prominent negative symptoms. METHOD: We adopted a mixed methods case series design, providing detailed quantitative data on changes over time, to focus on potential mechanisms underlying the intervention, in combination with qualitative interviews. RESULTS: The intervention showed good feasibility as demonstrated by the attendance rate, the positive feedback from participants and the multidisciplinary team, and the improvements on negative symptoms observed following the intervention. Multilevel modelling showed that depression, internalized stigma and reflective functioning explained the variance in negative symptoms. DISCUSSION: The pilot study indicated that the intervention has high feasibility and that improvements in negative symptoms can be partially explained by improvements on depression, stigma and reflective functioning.

Unpacking stigma: Meta-analyses of correlates and moderators of personal stigma in psychosis.

Personal stigma entails perceived, experienced and internalised stigmatisation. Mental Health stigma has been widely researched across a range of countries and a meta-analysis of their associations and moderators in psychosis is timely. Meta-analyses were conducted examining the correlates and moderators of personal stigma in terms of: (1) demographic variables (2) illness related variables (3) symptoms/negative outcomes, and (4) aspects of wellbeing. Associations were obtained from a total of 216 records. Several demographic factors including age, economic status, employment, and rural residence had small associations with aspects of personal stigma (r's = 0.12 to -0.13). Personal stigma aspects were inversely related to medication adherence (r's = -0.20, -0.21), and positively associated with insight and number of hospitalisations (r's = 0.09-0.19). Most symptoms were positively associated with personal stigma (r's = 0.10-0.43), whereas inverse relations with wellbeing variables were identified (r's = -0.13 to -0.54). Moderator effects emerged including that of cultural setting and sex, age and education level, highlighting the role of cultural and demographic factors in shaping personal stigma aspects in psychosis. The present study also highlights the importance of recognizing the negative effect of actual stigma and discrimination experiences; particularly its detrimental impact on self-image and its complex role in shaping the internalisation of societal stigma.

Fluorescence Spectroscopy Analysis of the Bacteria-Mineral Interface: Adsorption of Lipopolysaccharides to Silica and Alumina.

We present here a quantification of the sorption process and molecular conformation involved in the attachment of bacterial cell wall lipopolysaccharides (LPSs), extracted from Escherichia coli, to silica (SiO2) and alumina (Al2O3) particles. We propose that interfacial forces govern the physicochemical interactions of the bacterial cell wall with minerals in the natural environment, and the molecular conformation of LPS cell wall components depends on both the local charge at the point of binding and hydrogen bonding potential. This has an effect on bacterial adaptation to the host environment through adhesion, growth, function, and ability to form biofilms. Photophysical techniques were used to investigate adsorption of fluorescently labeled LPS onto mineral surfaces as model systems for bacterial attachment. Adsorption of macromolecules in dilute solutions was studied as a function of pH and ionic strength in the presence of alumina and silica via fluorescence, potentiometric, and mass spectrometry techniques. The effect of silica and alumina particles on bacterial growth as a function of pH was also investigated using spectrophotometry. The alumina and silica particles were used to mimic active sites on the surface of clay and soil particles, which serve as a point of attachment of bacteria in natural systems. It was found that LPS had a high adsorption affinity for Al2O3 while adsorbing weakly to SiO2 surfaces. Strong adsorption was observed at low pH for both minerals and varied with both pH and mineral concentration, likely in part due to conformational rearrangement of the LPS macromolecules. Bacterial growth was also enhanced in the presence of the particles at low pH values. This demonstrates that at a molecular level, bacterial cell wall components are able to adapt their conformation, depending on the solution pH, in order to maximize attachment to substrates and guarantee community survival.

The pH-responsive behaviour of poly(acrylic acid) in aqueous solution is dependent on molar mass.

Fluorescence spectroscopy on a series of aqueous solutions of poly(acrylic acid) containing a luminescent label showed that polymers with molar mass, Mn < 16.5 kDa did not exhibit a pH responsive conformational change, which is typical of higher molar mass poly(acrylic acid). Below this molar mass, polymers remained in an extended conformation, regardless of pH. Above this molar mass, a pH-dependent conformational change was observed. Diffusion-ordered nuclear magnetic resonance spectroscopy confirmed that low molar mass polymers did not undergo a conformational transition, although large molar mass polymers did exhibit pH-dependent diffusion.

Adsorption of poly acrylic acid onto the surface of calcite: an experimental and simulation study.

Macromolecular binding to minerals is of great importance in the formation of biofilms, and carboxylate functional groups have been found to play a pivotal role in the functioning of these macromolecules. Here we present both fluorescence time-resolved anisotropy measurements and simulation data on the conformational behaviour and binding of a poly acrylic acid polymer. In solution the polymer exhibits a pH dependent behaviour, with a coiled conformation at a low pH and extended conformation at higher pH values. The polymer is readily adsorbed on the surface of calcite, preferring to bind in an extended conformation, with the strength of the adsorption dependent on the pH and presence of counter ions. We discuss the reasons why the calculated adsorption free energy differs from that obtained from a Langmuir isotherm analysis, showing that they refer to different quantities. The enhanced binding of the extended conformations shows the importance of flexibility in the binding of macromolecules.

Förster resonance energy transfer confirms the bacterial-induced conformational transition in highly-branched poly(N-isopropyl acrylamide with vancomycin end groups on binding to Staphylococcus aureus.

We describe a series of experiments designed to investigate the conformational transition that highly-branched polymers with ligands undergo when interacting with bacteria, a process that may provide a new sensing mechanism for bacterial detection. Fluorescent highly-branched poly(N-isopropyl acrylamide)s (HB-PNIPAM) were prepared by sequential self-condensing radical copolymerizations, using anthrylmethyl methacrylate (AMMA) and fluorescein-O-acrylate (FA) as fluorescent comonomers and 4-vinylbenzyl pyrrole carbodithioate as a branch forming monomer. Differences in reactivity necessitated to first copolymerize AMMA then react with FA in a separate sequential monomer feed step. Modifications of the chain ends produced vancomycin-functional derivatives (HB-PNIPAM-Van). The AMMA and FA labels allow probing of the conformational behaviour of the polymers in solution via Förster resonance energy transfer experiments. It was shown that interaction of this polymer's end groups with Staphylococcus aureus induced a macromolecular collapse. The data thus provide conclusive evidence for a conformational transition that is driven by binding to a bacterium.

Neutralizing mutations of carboxylates that bind metal 2 in T5 flap endonuclease result in an enzyme that still requires two metal ions.

Flap endonucleases (FENs) are divalent metal ion-dependent phosphodiesterases. Metallonucleases are often assigned a "two-metal ion mechanism" where both metals contact the scissile phosphate diester. The spacing of the two metal ions observed in T5FEN structures appears to preclude this mechanism. However, the overall reaction catalyzed by wild type (WT) T5FEN requires three Mg(2+) ions, implying that a third ion is needed during catalysis, and so a two-metal ion mechanism remains possible. To investigate the positions of the ions required for chemistry, a mutant T5FEN was studied where metal 2 (M2) ligands are altered to eliminate this binding site. In contrast to WT T5FEN, the overall reaction catalyzed by D201I/D204S required two ions, but over the concentration range of Mg(2+) tested, maximal rate data were fitted to a single binding isotherm. Calcium ions do not support FEN catalysis and inhibit the reactions supported by viable metal cofactors. To establish participation of ions in stabilization of enzyme-substrate complexes, dissociation constants of WT and D201I/D204S-substrate complexes were studied as a function of [Ca(2+)]. At pH 9.3 (maximal rate conditions), Ca(2+) substantially stabilized both complexes. Inhibition of viable cofactor supported reactions of WT, and D201I/D204S T5FENs was biphasic with respect to Ca(2+) and ultimately dependent on 1/[Ca(2+)](2). By varying the concentration of viable metal cofactor, Ca(2+) ions were shown to inhibit competitively displacing two catalytic ions. Combined analyses imply that M2 is not involved in chemical catalysis but plays a role in substrate binding, and thus a two-metal ion mechanism is plausible.

Highly branched polymers with polymyxin end groups responsive to Pseudomonas aeruginosa.

Polymyxin peptide conjugated to the end groups of highly branched poly(N-isopropyl acrylamide) was shown to bind to a Gram negative bacterium, Pseudomonas aeruginosa . The nonbound polymer had a lower critical solution temperature (LCST) above 60 °C. However, binding caused aggregation, which was disrupted on cooling of the bacteria and polymer mixture. The data indicate that polymer binding of bacteria occurred by interaction of the end groups with lipopolysaccharide and that the binding decreased the LCST to below 37 °C. Cooling then progressed the polymer/bacteria aggregate through a bound LCST into an open polymer coil conformation that was not adhesive to P. aeruginosa .

Hyperbranched poly(NIPAM) polymers modified with antibiotics for the reduction of bacterial burden in infected human tissue engineered skin.

The escalating global incidence of bacterial infection, particularly in chronic wounds, is a problem that requires significant improvements to existing therapies. We have developed hyperbranched poly(NIPAM) polymers functionalized with the antibiotics Vancomycin and Polymyxin-B that are sensitive to the presence of bacteria in solution. Binding of bacteria to the polymers causes a conformational change, resulting in collapse of the polymers and the formation of insoluble polymer/bacteria complexes. We have applied these novel polymers to our tissue engineered human skin model of a burn wound infected with Pseudomonas aeruginosa and Staphylococcus aureus. When the polymers were removed from the infected skin, either in a polymer gel solution or in the form of hydrogel membranes, they removed bound bacteria, thus reducing the bacterial load in the infected skin model. These bacteria-binding polymers have many potential uses, including coatings for wound dressings.

Binding bacteria to highly branched poly(N-isopropyl acrylamide) modified with vancomycin induces the coil-to-globule transition.

Binding of highly branched poly(N-isopropylacrylamide) with vancomycin end groups to Staphylococcus aureus induced a coil-to-globule phase transition. The polymers aggregated this gram-positive bacteria (but not gram-negative bacteria) over a wide range of temperatures, but cooling to 24-26 degrees C progressed the polymer-bound bacteria through a globule-to-coil phase transition, after which the bacteria were released.

Development of three-dimensional tissue-engineered models of bacterial infected human skin wounds.

While infected skin wounds are on the increase because of ageing populations, rising incidence of diabetes, and antibiotic resistance, we lack relevant in vivo or in vitro models to study many aspects of bacterial interaction with skin. The aim of this study was to develop three-dimensional models of normal human skin to study bacterial infection. The common dermatological pathogens Staphylococcus aureus and Pseudomonas aeruginosa were used to infect tissue-engineered skin, and the course of infection in the skin was examined over several days. Two forms of model were developed-one in which bacteria were introduced directly to 10 mm wounds in the epidermis, and another in which wounds were created by burning a 4 mm hole in the center of the tissue before inoculation. The bacteria flourished within the engineered skin, and colonized the upper epidermal layers before invasion into the dermis. Infection with P. aeruginosa caused a loss of epidermis and de-keratinization of the skin constructs, as well as partial loss of basement membrane. These novel complex human skin infection models could be used to investigate microbial invasion of normal skin epithelium, basement membrane, and connective tissue, and as a model to study approaches to reduce bacterial burden in skin wounds.

The polymer physics and chemistry of microbial cell attachment and adhesion.

The attachment of microbial cells to solid substrata is a primary ecological strategy for the survival of species and the development of specific activity and function within communities. An hypothesis arising from a biological sciences perspective may be stated as follows: The attachment of microbes to interfaces is controlled by the macromolecular structure of the cell wall and the functional genes that are induced for its biological synthesis. Following logically from this is the view that diverse attached cell behaviour is mediated by the physical and chemical interactions of these macromolecules in the interfacial region and with other cells. This aspect can be reduced to its simplest form by treating physico-chemical interactions as colloidal forces acting between an isolated cell and a solid or pseudo solid substratum. These forces can be analysed by established methods rooted in DLVO (Derjaguin, Landau, Verwey and Overbeek) theory. Such a methodology provides little insight into what governs changes in the behaviour of the cell wall attached to surfaces, or indeed other cells. Nor does it shed any light on the expulsion of macromolecules that modify the interface such as formation of slime layers. These physical and chemical problems must be treated at the more fundamental level of the structure and behaviour of the individual components of the cell wall, for example biosurfactants and extracellular polysaccharides. This allows us to restate the above hypothesis in physical sciences terms: Cell attachment and related cell growth behaviour is mediated by macromolecular physics and chemistry in the interfacial environment. Ecological success depends on the genetic potential to favourably influence the interface through adaptation of the macromolecular structure, We present research that merges these two perspectives. This is achieved by quantifying attached cell growth for genetically diverse model organisms, building chemical models that capture the variations in interfacial structure and quantifying the resulting physical interactions. Experimental observations combine aqueous chemistry techniques with surface spectroscopy in order to elucidate the cell wall structure. Atomic force microscopy methods quantify the physical interactions between the solid substrata and key components of the cell wall such as macromolecular biosurfactants. Our current approach focuses on considering individually mycolic acids or longer chain polymers harvested from cells, as well as characterised whole cells. This approach allows us to use a multifactorial approach to address the relative impact of the individual components of the cell wall in contact with model surfaces. We then combine these components to increase complexity step-wise, while comparing with the behaviour of entire cells. Eventually, such an approach should allow us to estimate and understand the primary factors governing microbial cell adhesion. Although the work addresses the cell-mineral interface at a fundamental level, the research is driven by a range of technology needs. The initial rationale was improved prediction of contaminant degradation in natural environments (soils, sediments, aquifers) for environmental cleanup. However, this area of research addresses a wide range of biotechnology areas including improved understanding of pathogen survival (e.g., in surgical environments), better process intensification in biomanufacturing (biofilm technologies) and new product development.

Highly branched poly-(N-isopropylacrylamide)s with arginine-glycine-aspartic acid (RGD)- or COOH-chain ends that form sub-micron stimulus-responsive particles above the critical solution temperature.

Highly branched poly(-isopropyl acrylamide)s with peptide-end groups form colloidally stable dispersions of sub-micron particles above the lower critical solution temperature.

Highly branched poly(N-isopropylacrylamide) for use in protein purification.

Poly(N-isopropylacrylamide)s with imidazole endgroups were used to separate a histidine-tagged protein fragment directly from a crude cell lysate. The polymers display a lower critical solution temperature that can be tuned to occur at a range of subambient temperatures. UV-visible spectra indicated differences in the binding in aqueous media of Cu(II) and Ni(II) to the imidazole endgroups. These changes in the UV-visible spectra were reflected in the solution/aggregation behavior of the polymers as studied by dynamic light scattering. The addition of Cu(II) disaggregated the polymers, and the polymer coil swelled. On the other hand, when Ni(II) was added the polymers remained aggregated in aqueous media. The polymers were used to purify residues 230-534 of the histidine-tagged breast cancer susceptibility protein his6-BRCA1. Cu(II) was found to be better suited to the formation of useful polymer-metal ion-protein complexes that display cloud points, since Ni(II)/polymer mixtures generated very little purified protein. The polymers were synthesized using a previously reported variation of the reversible addition-fragmentation chain termination (RAFT) methodology, using the chain transfer agent 3H-imidazole-4-carbodithioic acid 4-vinyl benzyl ester with N-isopropylacrylamide (NIPAM).

Dinuclear monointercalating RuII complexes that display high affinity binding to duplex and quadruplex DNA.

The DNA duplex binding properties of previously reported dinuclear Ru(II) complexes based on the ditopic ligands tetrapyrido[3,2-a:2',3'-c:3'',2''-h:2'',3''-j]phenazine (tppz) and tetraazatetrapyrido[3,2-a:2'3'-c:3'',2''-l:2''',3'''-n]pentacene (tatpp) are reported. Photophysical and biophysical studies indicate that, even at high ionic strengths, these complexes bind to duplex DNA, through intercalation, with affinities that are higher than any other monointercalating complex and are only equalled by DNA-threaded bisintercalating complexes. Additional studies at high ionic strengths using the 22-mer d(AG(3)[T(2)AG(3)](3)) [G3] human telomeric sequence reveal that the dinuclear tppz-based systems also bind with high affinity to quadruplex DNA. Furthermore, for these complexes, quadruplex binding is accompanied by a distinctive blue-shifted "light-switch" effect, characterized by higher emission enhancements than those observed in the analogous duplex effect. Calorimetry studies reveal that the thermodynamics of duplex and quadruplex binding is distinctly different, with the former being entirely entropically driven and the latter being both enthalpically and entropically favored.

Complexes of substituted derivatives of 2-(2-pyridyl)benzimidazole with Re(I), Ru(II) and Pt(II): structures, redox and luminescence properties.

N,N'-Chelating ligands based on the 2-(2-pyridyl)benzimidazole (PB) core have been prepared with a range of substituents (phenyl, pentafluorophenyl, naphthyl, anthracenyl, pyrenyl) connected to the periphery via alkylation of the benzimidazolyl unit at one of the N atoms. These PB ligands have been used to prepare a series of complexes of the type [Re(PB)(CO)(3)Cl], [Pt(PB)(CCR)(2)](where -CCR is an acetylide ligand) and [Ru(bpy)(2)(PB)][PF(6)](2)(bpy = 2,2'-bipyridine). Six of the complexes have been structurally characterised. Electrochemical and luminescence studies show that all three series of complexes behave in a similar manner to the analogous complexes with 2,2'-bipyridine in place of PB. In particular, all three series of complexes show luminescence in the range 553-605 nm (Pt series), 620-640 nm (Re series) and 626-645 nm (Ru series) arising from the (3)MLCT state, with members of the Pt(II) series being the most strongly emissive with lifetimes of up to 500 ns and quantum yields of up to 6% in air-saturated CH(2)Cl(2) at room temperature. In the Re and Ru series there was clear evidence for inter-component energy-transfer processes in both directions between the (3)MLCT state of the metal centre and the singlet and triplet states of the pendant organic luminophores (naphthalene, pyrene, anthracene). For example the pyrene singlet is almost completely quenched by energy transfer to a Re-based MLCT excited state, which in turn is completely quenched by energy transfer to the lower-lying pyrene triplet state. For the analogous Ru(II) complexes the inter-component energy transfer is less effective, with (1)anthracene --> Ru((3)MLCT) energy transfer being absent, and Ru((3)MLCT)-->(3)anthracene energy transfer being incomplete. This is rationalised on the basis of a greater effective distance for energy transfer in the Ru(II) series, because the MLCT excited states are localised on the bpy ligands which are remote from the pendant aromatic group; in the Re series in contrast, the MLCT excited states involve the PB ligand to which the pendant aromatic group is directly attached, giving more efficient energy transfer.