Communication practices for delivering health behaviour change conversations in primary care: a systematic review and thematic synthesis.

BACKGROUND: Clinical guidelines exhort clinicians to encourage patients to improve their health behaviours. However, most offer little support on how to have these conversations in practice. Clinicians fear that health behaviour change talk will create interactional difficulties and discomfort for both clinician and patient. This review aims to identify how healthcare professionals can best communicate with patients about health behaviour change (HBC). METHODS: We included studies which used conversation analysis or discourse analysis to study recorded interactions between healthcare professionals and patients. We followed an aggregative thematic synthesis approach. This involved line-by-line coding of the results and discussion sections of included studies, and the inductive development and hierarchical grouping of descriptive themes. Top-level themes were organised to reflect their conversational positioning. RESULTS: Of the 17,562 studies identified through systematic searching, ten papers were included. Analysis resulted in 10 top-level descriptive themes grouped into three domains: initiating; carrying out; and closing health behaviour change talk. Of three methods of initiation, two facilitated further discussion, and one was associated with outright resistance. Of two methods of conducting behaviour change talk, one was associated with only minimal patient responses. One way of closing was identified, and patients did not seem to respond to this positively. Results demonstrated a series of specific conversational practices which clinicians use when talking about HBC, and how patients respond to these. Our results largely complemented clinical guidelines, providing further detail on how they can best be delivered in practice. However, one recommended practice - linking a patient's health concerns and their health behaviours - was shown to receive variable responses and to often generate resistance displays. CONCLUSIONS: Health behaviour change talk is smoothly initiated, conducted, and terminated by clinicians and this rarely causes interactional difficulty. However, initiating conversations by linking a person's current health concern with their health behaviour can lead to resistance to advice, while other strategies such as capitalising on patient initiated discussions, or collaborating through question-answer sequences, may be well received.

Occupational stress and coping mechanisms in crime scene personnel.

Background: Studies on occupational stress have shown that police officers (POs) are vulnerable to the effects of stress, demonstrated by increased risk of cardiometabolic diseases, which may be exacerbated by the use of maladaptive coping techniques. Although there is an abundance of research pertaining to stress in POs, little research has been done to assess a subset of law enforcement, crime scene personnel (CSP). Aims: To assess the stress levels, anxiety levels and coping mechanisms of CSP across the state of Texas. Methods: The Perceived Stress Scale (PSS), Police Stress Questionnaire (PSQ), and the Distress Thermometer were used to measure stress levels, the State-Trait Anxiety Inventory (STAI) was utilized to measure anxiety, and the Brief COPE questionnaire was used to measure coping mechanisms. Results: CSP (N = 76) surveyed reported both low stress and low anxiety for all measures used, with males reporting slightly higher stress and anxiety than females. Differences in coping mechanisms used by CSP were observed between males and females, but not between sworn officers and civilian workers. Female CSP used emotional support (P < 0.01), instrumental support (P < 0.05) and positive reframing (P < 0.05) as a coping mechanism significantly more often than males. Conclusions: The results suggest that adaptive coping mechanisms should be emphasized by those supervising CSP. With little research available on CSP, further evaluation of the type of stressors experienced by these members of law enforcement is warranted.

Exogenous Carbohydrate Reduces Cortisol Response from Combined Mental and Physical Stress.

Combined mental and physical stress is associated with exacerbated cortisol production which may increase risk for the progression of cardiovascular disease in individuals working in high-stress occupations (e.g., firefighters, military personnel, etc.). Carbohydrate (CHO) ingestion prior to physical stress may attenuate cortisol concentrations. This project was the first to investigate the effect of CHO ingestion on cortisol response from combined mental and physical stress. 16 men 21-30 years old were randomly assigned a 6.6% CHO beverage or non-CHO control 15 min prior to performing a dual-concurrent-stress challenge. This consisted of physical stress (i.e., steady state exercise) combined with computerized mental challenges. Blood was sampled 70, 40, and 15 min before exercise, immediately at onset of exercise, 10, 20, 30, 35 min during exercise, and 15, 30, 45, and 60 min after exercise. There was a significant main effect for treatment regarding mean cortisol concentrations (F=5.30, P=0.0219). The total area under curve for cortisol was less when CHO was ingested (T7=4.07, P=0.0048). These findings suggest that CHO ingestion immediately prior to combined mental and physical stress may attenuate cortisol responses.

Impact of an educational intervention upon the hand hygiene compliance of children.

BACKGROUND: Hand hygiene compliance is the single most effective way to reduce healthcare-associated infections. Children are notoriously vulnerable to infection as well as acting as conduits to transmission. Based on these observations, the authors formulated the hypothesis that behavioural change which improved children's hand hygiene compliance would decrease the spread of infectious diseases. AIM: To create an educational intervention to induce long-term behavioural change culminating in increased hand hygiene compliance of children, and thus a decrease in the rate of infections. METHODS: Focus groups conducted during interactive teaching sessions identified what children felt would help them to increase their hand hygiene compliance. This informed the design of an educational device that was subsequently trialled to measure its effectiveness in increasing hand hygiene compliance. Initial developmental stages were conducted in two schools in the East Midlands with study participants aged 5-8 years; the device was subsequently used in a healthcare setting to assess deployment flexibility. FINDINGS: Focus groups indicated that children enjoyed interactive learning, developed knowledge about cross-transmission of infections, and became motivated to encourage others to improve hand hygiene compliance. Microbiological swabbing verified the presence of pathogens on children's hands and environmental surfaces that could serve as reservoirs of infection, and questionnaires indicated an increase in handwashing following the intervention. CONCLUSION: Educational interventions have the potential to increase hand hygiene and reduce the transmission of infections.

Changing trends in HIV diagnosis in an inner city London teaching hospital 2007-2011.

In 2008, guidelines aimed at reducing undiagnosed HIV infection were published, recommending normalization of testing. Local initiatives were carried out in south-west London including an extensive primary care education programme. This study aimed to establish the impact of these initiatives on HIV diagnoses. Data on all new diagnoses referred to our centre were collected and analysed from patient records. A total of 394 patients were newly diagnosed with HIV from 2007 to 2011. Diagnosis in primary care and other non-sexual health (SH) settings increased by 184% during this period. There was an increase of 37% in the median CD4 count at diagnosis, taking the figure to 372 cells/mm³ in 2011. The proportion of patients presenting with a CD4 count of <200cells/mm³ decreased by 24%. The evidence suggests that these local initiatives were successful in increasing HIV diagnoses outside SH settings, particularly in very late presenters.

Bacterial attachment on sub-nanometrically smooth titanium substrata.

Despite the volume of work that has been conducted on the topic, the role of surface topography in mediating bacterial cell adhesion is not well understood. The primary reason for this lack of understanding is the relatively limited extent of topographical characterisation employed in many studies. In the present study, the topographies of three sub-nanometrically smooth titanium (Ti) surfaces were comprehensively characterised, using nine individual parameters that together describe the height, shape and distribution of their surface features. This topographical analysis was then correlated with the adhesion behaviour of the pathogenic bacteria Staphylococcus aureus and Pseudomonas aeruginosa, in an effort to understand the role played by each aspect of surface architecture in influencing bacterial attachment. While P. aeruginosa was largely unable to adhere to any of the three sub-nanometrically smooth Ti surfaces, the extent of S. aureus cell attachment was found to be greater on surfaces with higher average, RMS and maximum roughness and higher surface areas. The cells also attached in greater numbers to surfaces that had shorter autocorrelation lengths and skewness values that approached zero, indicating a preference for less ordered surfaces with peak heights and valley depths evenly distributed around the mean plane. Across the sub-nanometrically smooth range of surfaces tested, it was shown that S. aureus more easily attached to surfaces with larger features that were evenly distributed between peaks and valleys, with higher levels of randomness. This study demonstrated that the traditionally employed amplitudinal roughness parameters are not the only determinants of bacterial adhesion, and that spatial parameters can also be used to predict the extent of attachment.

Air-directed attachment of coccoid bacteria to the surface of superhydrophobic lotus-like titanium.

Superhydrophobic titanium surfaces fabricated by femtosecond laser ablation to mimic the structure of lotus leaves were assessed for their ability to retain coccoid bacteria. Staphylococcus aureus CIP 65.8T, S. aureus ATCC 25923, S. epidermidis ATCC 14990T and Planococcus maritimus KMM 3738 were retained by the surface, to varying degrees. However, each strain was found to preferentially attach to the crevices located between the microscale surface features. The upper regions of the microscale features remained essentially cell-free. It was hypothesised that air entrapped by the topographical features inhibited contact between the cells and the titanium substratum. Synchrotron SAXS revealed that even after immersion for 50 min, nano-sized air bubbles covered 45% of the titanium surface. After 1 h the number of cells of S. aureus CIP 65.8T attached to the lotus-like titanium increased to 1.27×10(5) mm(-2), coinciding with the replacement of trapped air by the incubation medium.

High rate of postoperative mortality in patients with mucopolysaccharidosis I: findings from the MPS I Registry.

BACKGROUND/PURPOSE: Mucopolysaccharidosis I (MPS I) is a rare lysosomal storage disorder caused by deficiency of α-L-iduronidase, which results in progressive multisystemic disease. Patients with MPS I often require multiple common and uncommon surgeries and are at risk for surgical and anesthetic complications because of respiratory and cardiac disease. Surgery often precedes diagnosis; thus, surgeons and anesthesiologists may be unaware of potential risks. METHODS: We analyzed data from the MPS I Registry, a voluntary observational database, for deaths occurring within 1 month of a surgical procedure among the 932 patients enrolled as of July 2010. RESULTS: Among the 196 deceased patients, 186 reported 1 surgery or more, and 32 had 1 surgery or more within 1 month of death, including 20 who had 1 surgery or more within 10 days of death. Surgeries before death included hernia repair, central line placement, spinal surgery, tracheostomy, and ventriculo-peritoneal shunt. Most patients (28/32) had severe MPS I (Hurler), and 20 of 32 patients (all Hurler) died at 3 years or younger. In 6 of 32 patients, surgery was directly noted in the cause of death, including 4 patients with an attenuated form of MPS I. CONCLUSIONS: Patients with mucopolysaccharidosis have a high postoperative mortality because of underlying respiratory and cardiac diseases.

Experimental jetlag disrupts circadian clock genes but improves performance in racehorses after light-dependent rapid resetting of neuroendocrine systems and the rest-activity cycle.

Abrupt alterations in the 24-h light : dark cycle, such as those resulting from transmeridian air travel, disrupt circadian biological rhythms in humans with detrimental consequences on cognitive and physical performance. In the present study, a jetlag-simulated phase shift in photoperiod temporally impaired circadian peaks of peripheral clock gene expression in racehorses but acutely enhanced athletic performance without causing stress. Indices of aerobic and anaerobic capacities were significantly increased by a phase-advance, enabling prolonged physical activity before fatigue occurred. This was accompanied by rapid re-entrainment of the molecular clockwork and the circadian pattern of melatonin, with no disturbance of the adrenal cortical axis, but a timely rise in prolactin, which is a hormone known to target organs critical for physical performance. Subsequent studies showed that, unlike the circadian pattern of melatonin, and in contrast to other species, the daily rhythm of locomotor activity was completely eliminated under constant darkness, but it was restored immediately upon the reintroduction of a light : dark cycle. Resetting of the rhythm of locomotion was remarkably fast, revealing a rapid mechanism of adaptation and a species dependency on light exposure for the expression of daily diurnal activity. These results show that horses are exquisitely sensitive to sudden changes in photoperiod and that, unlike humans, can benefit from them; this appears to arise from powerful effects of light underlying a fast and advantageous process of adjustment to the phase shift.

Nature inspired structured surfaces for biomedical applications.

Nature has created an array of superhydrophobic surfaces that possess water-repellent, self-cleaning and anti-icing properties. These surfaces have a number of potential applications in the biomedical industry, as they have the potential to control protein adsorption and cell adhesion. Natural superhydrophobic surfaces are typically composed of materials with a low intrinsic surface free-energy (e.g the cuticular waxes of lotus leaves and insect wings) with a hierarchical structural configuration. This hierarchical surface topography acts to decrease the contact area of water droplets in contact with the surface, thereby increasing the extent of the air/water interface, resulting in water contact angles greater than 150º. In order to employ these surfaces in biotechnological applications, fabrication techniques must be developed so that these multi-scale surface roughness characteristics can be reproduced. Additionally, these fabrication techniques must also be able to be applied to the material required for the intended application. An overview of some of the superhydrophobic surfaces that exist in nature is presented, together with an explanation of the theories of their wettability. Also included is a description of some of the biomedical applications of superhydrophobic surfaces and fabrication techniques that can be used to mimic superhydrophobic surfaces found in nature.

Re-attendance to the emergency department.

The re-attendance of patients to emergency departments is becoming an increasingly common problem and yet there is not much research available on this subject. This report describes an audit carried out in the emergency department of this hospital, specifically looking at the issue of re-attendance and if there is anything that can be done to prevent it. Traditionally, emergency departments in the United Kingdom have been staffed at junior doctor level by senior house officers (SHO); however, with the advent of modernising medical careers in the United Kingdom, departments are being staffed at this level by foundation year 2 doctors (FY2). Whereas SHO range in experience from a year post-qualification to a possible 5-6 years post-qualification, all FY2 doctors have only one year post-qualification clinical experience. In this audit, FY2 doctors, despite seeing fewer patients per head than the middle-grade tier, had a higher level of re-attendance and it is hypothesised whether this is due to the impact of modernising medical careers on their training.

Cardiorespiratory responses of Hi Fit and Low Fit subjects to mental challenge during exercise.

The influence of psychological states on physiological responses during exercise is of considerable importance to individuals for which the efficiency of energy production is critical to occupational performance. Numerous studies have shown that aerobic fitness is associated with enhanced cardiovascular efficiency at rest and that responses to mental stress demonstrate evidence of increased sensitivity (relative increase in HR response) and enhanced efficiency (a decrease in absolute HR). However, the effect of aerobic fitness and its impact on cardiorespiratory (CR) responses to psychological stress during exercise has not been investigated. Therefore, the purpose of this study was three-fold; (1) to examine during exercise, anxiety, effort sense, and CR responses to a mental challenge, (2) to examine anxiety and heart rate (HR) responses from rest to exercise with mental challenge between below average fitness (Low Fit) and well-above average fitness (Hi Fit) individuals (exercising at similar relative intensities), and (3) to examine anxiety, effort sense, and CR responses of Low Fit and Hi Fit individuals to a mental challenge during exercise at a similar relative intensity. Twelve Low Fit and eleven Hi Fit subjects participated in two, 32-minute cycle ergometer rides at 65 % of VO2max. In the mental challenge condition (MCC), subjects rode while participating in mentally challenging tasks (Stroop Color-Word task and mental arithmetic) from min 6 to min 14 of the protocol. In the no mental challenge condition (NMCC), subjects exercised at the same intensity and duration without a stressor. Subjects were counter-balanced between fitness levels and condition. HR, VE, VE/VO2, RR, VO2, RER, effort sense (RPE), and state anxiety (SAI) were assessed at 5, 14, 24, and 30 min. SAI was also assessed at - 5 min before exercise and after 15 min of recovery. In addition, the NASA task load index (NTLX) was used to assess perceived overall workload. SAI increased significantly at 14 min in the MCC. NTLX scores indicated that the MCC was perceived as a greater overall workload. Furthermore, HR, VE, VE/VO2, and RR were significantly elevated during the mental challenge condition at 14 min. The Hi Fit subjects tended to respond to the dual stress of exercise and mental challenge with a relative increase in HR, while absolute HR was similar in both groups. An examination of fitness group differences revealed that SAI and NTLX were similar for Low Fit and Hi Fit subjects when exercising in the MCC, although, Hi Fit subjects demonstrated lower HR responses from 6 min to 14 min. VE, VE/VO2, and RR were similar for Low Fit and Hi Fit subjects. These results suggest that psychological stress during physical activity can exacerbate cardiorespiratory responses and suggests that factors that impact CR adjustment to mental challenge from resting baseline may differ from the factors that impact CR adjustment to mental challenge during exercise. Finally, fitness level attenuates HR and may attenuate additional cardiorespiratory responses while participating in a dual stress condition, of exercise and mental challenge.

EBV EBNA 2 stimulates CDK9-dependent transcription and RNA polymerase II phosphorylation on serine 5.

EBNA 2 is one of only five viral genes essential for the infection and immortalization of human B cells by the cancer-associated virus Epstein-Barr virus (EBV). EBNA 2 activates cellular and viral transcription and associates with components of the basal transcription apparatus and a number of coactivators. We provide the first evidence to show that the mechanism of transcriptional activation by EBNA 2 also involves phosphorylation of the C-terminal domain (CTD) of RNA polymerase II (pol II). We found that transcriptional activation by EBNA 2 was inhibited by a dominant-negative mutant of the pol II CTD kinase, CDK9, and by low concentrations of the CDK9 inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole. Moreover, using chromatin immunoprecipitation assays we demonstrated that EBNA 2 stimulates both pol II recruitment and pol II phosphorylation on serine 5 of the CTD in vivo. These results identify a new step in the transcription cycle that is subject to regulation by a key EBV-encoded transcription factor and highlight CDK9 inhibitors as potential anti-EBV agents.

Employment of broad-range 16S rRNA PCR to detect aetiological agents of infection from clinical specimens in patients with acute meningitis--rapid separation of 16S rRNA PCR amplicons without the need for cloning.

AIMS: The aim of this study was to develop a polyacrylamide gel electrophoresis (PAGE) method for the rapid separation of 16S rRNA PCR amplicons from aetiological agents of acute meningitis. METHODS AND RESULTS: Blood samples from 40 patients with suspected acute meningococcal meningitis were examined for the presence of causal agents, including Neisseria meningitidis employing two methods: (i) broad-range 16S rRNA PCR in conjunction with PAGE and automated sequencing and (ii) species-specific PCR employing ABI TaqMan technology for N. meningitidis. Analysis of clinical specimens employing 16S rRNA PCR yielded 33/40 (82.5%) positive for the presence of bacterial DNA. Species-specific PCR yielded 30/40 (75%) clinical specimens positive for N. meningitidis. Prior to separation by PAGE, only 6/33 (18.2%) amplicons were able to be identified by sequence analysis, the remaining amplicons (n=27) did not yield an identification due to the presence of mixed 16S rRNA PCR amplicons. Following separation, amplicons were re-amplified and sequenced, yielding 24/27 (88.9%) positive for N. meningitidis and three specimens positive for Acinetobacter sp., Staphylococcus aureus and Streptococcus pneumoniae. One specimen was positive for both N. meningitidis and Streptococcus spp. and another specimen was positive for N. meningitidis and Pseudomonas sp., by broad-range PCR. Seven clinical specimens were negative for N. meningitidis and other eubacteria using both detection techniques. CONCLUSIONS: Clinical specimens including blood and cerebrospinal fluid from patients with suspected acute bacterial meningitis, may become contaminated with commensal skin flora, resulting in difficulties in downstream sequencing of pathogen plus contaminant DNA. This study allows for the rapid separation of amplified pathogen from contaminant DNA. SIGNIFICANCE AND IMPACT OF STUDY: This study demonstrated the usefulness of the rapid separation of multiple 16S rRNA PCR amplicons using a combination of PAGE and automated sequencing, without the need of cloning. Adoption of this technique is therefore proposed when trying to rapidly identify pathogens in clinical specimens employing broad-range 16S rRNA PCR.

Eradication of endotracheal tube biofilm by nebulised gentamicin.

OBJECTIVE: To compare the efficacy of gentamicin, nebulised via the endotracheal tube (ET), with that of parenteral cefotaxime or parenteral cefuroxime in preventing the formation of ET biofilm. SETTING: General intensive care units in two university teaching hospitals. DESIGN: The microbiology of ET biofilm from 36 ICU patients eligible to receive antibiotic prophylaxis was examined. Peak and trough tracheal concentrations of gentamicin, cefotaxime or cefuroxime were measured in each patient group, on the 2nd day of intubation. PATIENTS: Twelve patients received gentamicin (80 mg) nebulised in 4 ml normal saline every 8 h, 12 cefotaxime (1 g, 12 hourly) and 12 cefuroxime (750 mg, 8 hourly). Prophylaxis was continued for the duration of intubation. MEASUREMENTS AND RESULTS: Samples of tracheal secretions were taken on the 2nd day of ventilation for determination of antibiotic concentrations. Following extubation, ETs were examined for the presence of biofilm. Pathogens considered to be common aetiological agents for VAP included Staphylococcus aureus, enterococci, Enterobacteriaceae and pseudomonads. While microbial biofilm was found on all ETs from the cephalosporin group, microbial biofilm of these micro-organisms was found on 7 of the 12 ET tubes from patients receiving cefotaxime [ S. aureus (4), pseudomonads (1), Enterobacteriaceae (1), enterococcus (1)] and 8 of the 12 ET tubes from patients receiving cefuroxime [Enterobacteriaceae (6), P. aeruginosa (1) and enterococcus (1)]. While microbial biofilm was observed on five ETs from patients receiving nebulised gentamicin, none of these were from pathogens for ventilator-associated pneumonia (VAP). Tracheal concentrations of both cephalosporins were lower than those needed to inhibit the growth of pathogens recovered from ET tube biofilm. The median (and range) concentrations for cefotaxime were 0.90 (<0.23-1.31) mg/l and 0.28 (<0.23-0.58) mg/l for 2 h post-dose and trough samples, respectively. Two hours post-dose concentrations of cefuroxime (median and range) were 0.40 (0.34-0.83) mg/l, with trough concentrations of 0.35 (<0.22-0.47) mg/l. Tracheal concentrations (median and range) of gentamicin measured 1 h post-nebulisation were 790 (352-->1250) mg/l and then, before the next dose, were 436 (250-1000) mg/l. CONCLUSION: Nebulised gentamicin attained high concentrations in the ET lumen and was more effective in preventing the formation of biofilm than either parenterally administered cephalosporin and therefore may be effective in preventing this complication of mechanical ventilation.

Amino acid/spermine conjugates: polyamine amides as potent spermidine uptake inhibitors.

In this paper we describe the synthesis and characterization of a series of simple spermine/amino acid conjugates, some of which potently inhibit the uptake of spermidine into MDA-MB-231 breast cancer cells. The presence of an amide in the functionalized polyamine appeared to add to the affinity for the polyamine transporter. The extensive biological characterization of an especially potent analogue from this series, the Lys-Spm conjugate (31), showed this molecule will be an extremely useful tool for use in polyamine research. It was shown that the use of 31 in combination with DFMO led to a cytostatic growth inhibition of a variety of cancer cells, even when used in the presence of an extracellular source of transportable spermidine. It was furthermore shown that this combination effectively reduced the cellular levels of putrescine and spermidine while not affecting the levels of spermine. These facts together with the nontoxic nature of 31 make it a novel lead for further anticancer development.

Emergency medicine in Ecuador.

Emergency medical care in Ecuador is limited by geographic, economic, political, and infrastructural barriers. Afflictions of the developing world (eg, tropical infections and natural disasters) combine with ailments of the developed world (eg, trauma and cardiovascular disease) to mandate improved emergency medical systems. The nation has recently initiated FASBASE, a program dedicated to the enhancement of both prehospital and emergency department (ED) services. Furthermore, a dedicated residency program in Emergency and Disaster Medicine recently graduated its first class. Although more programs and funding are necessary to sustain the effort, Ecuador has begun to develop a modern emergency medical system.

Scandium(III) coordination polymers containing capsules based on two p-sulfonatocalix[4]arenes.

Reactions of sodium p-sulfonatocalix[4]arene and scandium(III) tristriflate in the presence, and absence, of [18]crown-6 give the crystalline complexes [Sc2(mu-OH)2(H2O)10][Na4(H2O)8-[calix[4]arene(SO3)4]2).13 H2O and [[Sc2(mu-OH)2(H2O)8][Sc(H2O)4]2[calix[4]-arene(SO3)4-H+]2([18]crown-6).16H2O. Both complexes involve novel coordination polymers with calixarene units linked through sodium or scandium centers and also feature capsule assemblies through to the head-to-head association of calixarenes. A linear array of capsules associated with an infinite chain of aquo-bridged sodium ions, and an aquated hydroxy-bridged scandium(III) dimer, [Sc2(mu-OH)2(H2O)10]4+, are found in the absence of the crown ether. In the presence of [18]crown-6 both hydrated scandium monomers and dimers bridge between calixarenes in a two-dimensional coordination network. The crown ethers reside in cavities created by two calixarenes from adjacent polymeric sheets via a variety of supramolecular interactions(hydrogen-bonding, shape complementarity), and effectively add a third dimension to the network. The extended structure of both of these polymers is highly porous, and resembles a bilayer.

Interaction of the periplasmic peptidylprolyl cis-trans isomerase SurA with model peptides. The N-terminal region of SurA id essential and sufficient for peptide binding.

One of the rate-limiting steps in protein folding has been shown to be the cis-trans isomerization of proline residues, which is catalyzed by a range of peptidylprolyl cis-trans isomerases. To characterize the interaction between model peptides and the periplasmic peptidylprolyl cis-trans isomerase SurA from E. coli, we employed a chemical cross-linking strategy that has been used previously to elucidate the interaction of substrates with other folding catalysts. The interaction between purified SurA and model peptides was significant in that it showed saturation and was abolished by denaturation of SurA; however the interaction was independent of the presence of proline residues in the model peptides. From results obtained by limited proteolysis we conclude that an N-terminal fragment of SurA, comprising 150 amino acids that do not contain the active sites involved in the peptidylprolyl cis-trans isomerization, is essential for the binding of peptides by SurA. This was confirmed by probing the interaction of the model peptide with the recombinant N-terminal fragment, expressed in Escherichia coli. Hence we propose that, similar to protein disulfide isomerase and other folding catalysts, SurA exhibits a modular architecture composed of a substrate binding domain and distinct catalytically active domains.

The coatomer of Trypanosoma brucei.

Coatomer is a multisubunit complex involved in trafficking of vesicles between the endoplasmatic reticulum and the Golgi apparatus. From sequence homologies, all seven subunits, alpha-, beta-, beta'-, gamma-, delta-, epsilon-, and zeta-COP, are encoded in the genome of Trypanosoma brucei. The complete predicted amino-acid sequences of beta-, beta'-, and zeta-COP show only 20-30% identity with higher eucaryotic homologues. The trypanosome coatomer complex was partially purified using a procedure similar to that used for bovine coatomer.