Health outcomes of patients with type 2 diabetes following bariatric surgery: Results from a publicly funded initiative.

OBJECTIVE: Bariatric surgery is an effective treatment for type 2 diabetes and morbid obesity. This paper analyses the clinical and patient-reported outcomes of patients treated through the Bariatric Surgery Initiative, a health system collaboration providing bariatric surgery as a state-wide public service in Queensland, Australia. RESEARCH DESIGN AND METHODS: A longitudinal prospective cohort study was undertaken. Eligible patients had type 2 diabetes and morbid obesity (BMI ≥ 35 kg/m2). Following referral by specialist outpatient clinics, 212 patients underwent Roux-en-Y gastric bypass or sleeve gastrectomy. Outcomes were tracked for a follow-up of 12-months and included body weight, BMI, HbA1c, comorbidities, health-related quality of life, eating behaviour, and patient satisfaction. RESULTS: Following surgery, patients' average body weight decreased by 23.6%. Average HbA1c improved by 24.4% and 48.8% of patients were able to discontinue diabetes-related treatment. The incidence of hypertension, non-alcoholic steatohepatitis, and renal impairment decreased by 37.1%, 66.4%, and 62.3%, respectively. Patients' emotional eating scores, uncontrolled eating and cognitive restraint improved by 32.5%, 20.7%, and 6.9%, respectively. Quality of life increased by 18.8% and patients' overall satisfaction with the treatment remained above 97.5% throughout the recovery period. CONCLUSIONS: This study confirmed previous work demonstrating the efficacy of publicly funded bariatric surgery in treating obesity, type 2 diabetes and related comorbidities, and improving patients' quality of life and eating behaviour. Despite the short follow-up period, the results bode well for future weight maintenance in this cohort.

Drosophila p38 MAPK interacts with BAG-3/starvin to regulate age-dependent protein homeostasis.

As organisms age, they often accumulate protein aggregates that are thought to be toxic, potentially leading to age-related diseases. This accumulation of protein aggregates is partially attributed to a failure to maintain protein homeostasis. A variety of genetic factors have been linked to longevity, but how these factors also contribute to protein homeostasis is not completely understood. In order to understand the relationship between aging and protein aggregation, we tested how a gene that regulates lifespan and age-dependent locomotor behaviors, p38 MAPK (p38Kb), influences protein homeostasis as an organism ages. We find that p38Kb regulates age-dependent protein aggregation through an interaction with starvin, a regulator of muscle protein homeostasis. Furthermore, we have identified Lamin as an age-dependent target of p38Kb and starvin.

Characterisation of trehalose-6-phosphate phosphatases from bacterial pathogens.

The trehalose biosynthesis pathway has recently received attention for therapeutic intervention combating infectious diseases caused by bacteria, helminths or fungi. Trehalose-6-phosphate phosphatase (TPP) is a key enzyme of the most common trehalose biosynthesis pathway and a particularly attractive target owing to the toxicity of accumulated trehalose-6-phosphate in pathogens. Here, we characterised TPP-like proteins from bacterial pathogens implicated in nosocomial infections in terms of their steady-state kinetics as well as pH- and metal-dependency of their enzymatic activity. Analysis of the steady-state kinetics of recombinantly expressed enzymes from Acinetobacter baumannii, Corynebacterium diphtheriae and Pseudomonas stutzeri yielded similar kinetic parameters as those of other reported bacterial TPPs. In contrast to nematode TPPs, the divalent metal ion appears to be bound only weakly in the active site of bacterial TPPs, allowing the exchange of the resident magnesium ion with other metal ions. Enzymatic activity comparable to the wild-type enzyme was observed for the TPP from P. stutzeri with manganese, cobalt and nickel. Analysis of the enzymatic activity of S. maltophilia TPP active site mutants provides evidence for the involvement of four canonical aspartate residues as well as a strictly conserved histidine residue of TPP-like proteins from bacteria in the enzyme mechanism. That histidine residue is a member of an interconnected network of five conserved residues in the active site of bacterial TPPs which likely constitute one or more functional units, directly or indirectly cooperating to enhance different aspects of the catalytic activity.

Trading Health for Wealth: The Effect of COVID-19 Response Stringency.

International governments' COVID-19 responses must balance human and economic health. Beyond slowing viral transmission, strict lockdowns have severe economic consequences. This work investigated response stringency, quantified by the Oxford COVID-19 Government Response Tracker's Stringency Index, and examined how restrictive interventions affected infection rates and gross domestic product (GDP) in China and OECD countries. Accounting for response timing, China imposed the most stringent restrictions, while Sweden and Japan were the least stringent. Expected GDP declines range from -8% (Japan) to -15.4% (UK). While greater restrictions generally slowed viral transmission, they failed to reach statistical significance and reduced GDP (p = 0.006). Timing was fundamental: governments who responded to the pandemic faster saw greater reductions in viral transmission (p = 0.013), but worse decreases in GDP (p = 0.044). Thus, response stringency has a greater effect on GDP than infection rates, which are instead affected by the timing of COVID-19 interventions. Attempts to mitigate economic impacts by delaying restrictions or decreasing stringency may buoy GDP in the short term but increase infection rates, the longer-term economic consequences of which are not yet fully understood. As highly restrictive interventions were successful in some but not all countries, decision-makers must consider whether their strategies are appropriate for the country on health and economic grounds.

Risk Factors for Postoperative Urinary Tract Infections Following Anterior Lumbar Interbody Fusion.

BACKGROUND: Although risk factors contributing to UTI have been studied in posterior approaches to lumbar fusion, there is a lack of literature on factors contributing to UTI in anterior lumbar interbody fusion (ALIF). Our purpose was to identify preoperative independent risk factors for postoperative urinary tract infection (UTI) following anterior lumbar interbody fusion (ALIF) so that surgeons may be able to initiate preventative measures and minimize the risk of UTI-related morbidity following ALIF. METHODS: The American College of Surgeons-National Surgical Quality Improvement Program database was queried to identify 10 232 patients who had undergone ALIF from 2005 to 2016; 144 patients (1.41%) developed a postoperative UTI while 10 088 patients (98.59%) did not. Univariate analyses were conducted to compare the 2 cohorts' demographics and preoperative comorbidities. Multivariate logistic regression models were then utilized to identify significant predictors of postoperative UTI following ALIF while controlling for differences seen in univariate analyses. RESULTS: Age ≥ 60 years (P = .022), female sex (P < .001), alcohol use (P = .014), open wound or wound infections (P = .019), and steroid use (P = .046) were independent risk factors for postoperative UTI. Longer operative times were also independent predictors for developing UTI: 120 minutes ≤ x < 180 minutes (P = .050), 180 minutes ≤ x < 240 minutes (P = .025), and ≥ 240 minutes (P = .001). Postoperative UTI independently increased the risk for pneumonia, blood transfusions, sepsis, thromboembolic events, and extended length of stay as well. CONCLUSIONS: Age ≥ 60 years, female sex, alcohol use, steroid use, and open wound or wound infections independently increased the risk for UTI following ALIF. Future work analyzing the efficacy of tapering alcohol and steroid use preoperatively and reducing procedural time with the aim of lowering UTI risk is warranted. Preoperative wound care is strongly encouraged to decrease UTI risk. LEVEL OF EVIDENCE: III.

A suicide inhibitor of nematode trehalose-6-phosphate phosphatases.

Protein-based drug discovery strategies have the distinct advantage of providing insights into the molecular mechanisms of chemical effectors. Currently, there are no known trehalose-6-phosphate phosphatase (TPP) inhibitors that possess reasonable inhibition constants and chemical scaffolds amenable to convenient modification. In the present study, we subjected recombinant TPPs to a two-tiered screening approach to evaluate several diverse compound groups with respect to their potential as TPP inhibitors. From a total of 5452 compounds tested, N-(phenylthio)phthalimide was identified as an inhibitor of nematode TPPs with apparent Ki values of 1.0 µM and 0.56 µM against the enzymes from the zoonotic roundworms Ancylostoma ceylanicum and Toxocara canis, respectively. Using site-directed mutagenesis, we demonstrate that this compound acts as a suicide inhibitor that conjugates a strictly conserved cysteine residue in the vicinity of the active site of nematode TPPs. The anthelmintic properties of N-(phenylthio)phthalimide were assessed in whole nematode assays using larvae of the ascaroids T. canis and T. cati, as well as the barber's pole worm Haemonchus contortus. The compound was particularly effective against each of the ascaroids with an IC50 value of 9.3 µM in the survival assay of T. cati larvae, whereas no bioactivity was observed against H. contortus.

Identification of a Novel Scaffold for Inhibition of Dipeptidyl Peptidase-4.

Dipeptidyl peptidase-4 (DPP-4) is considered a major drug target for type 2 diabetes mellitus (T2DM). In addition to T2DM, a regulatory role of DPP-4 was also found in cardiovascular diseases. Existing DPP-4 inhibitors have been reported to have several adverse effects. In this study, a computer-aided drug design approach and its use to detect a novel class of inhibitor for DPP-4 are reported. Through structure and pharmacophore-based screening, we identified 13 hit compounds from an ∼4-million-compound library. Physical interactions of these hits with DPP-4 were studied using docking and explicit solvent molecular dynamics (MD) simulations. Later, MMPBSA binding energy was calculated for the ligand/protein simulation trajectories to determine the stability of compounds in the binding cavity. These compounds have a novel scaffold and exhibited a stable binding mode. "Best-in-screen" compounds (or their closest available analogs) were resourced and their inhibition of DPP-4 activity was experimentally validated using an in vitro enzyme activity assay in the presence of 100 and 10 µM compounds. These assays identified a compound with a spirochromanone center with 53% inhibition activity at a 100 µM concentration. A further five spirochromanone compounds were synthesized and examined in silico and in vitro; again, one compound showed 53% inhibitory activity action at 100 µM. Overall, this study identified two novel "spirochromanone" compounds that lowered DPP-4 activity by more than ∼50% at 100 µM. This study also showed the impact of fast in silico drug design techniques utilizing virtual screening and MD to identify novel scaffolds to bind and inhibit DPP-4. Spirochromanone motif identified here may be used to design molecules to achieve drug-like inhibitory action against DPP-4.

DRfit: a Java tool for the analysis of discrete data from multi-well plate assays.

BACKGROUND: Analyses of replicates in sets of discrete data, typically acquired in multi-well plate formats, is a recurring task in many contemporary areas in the Life Sciences. The availability of accessible cross-platform data analysis tools for such fundamental tasks in varied projects and environments is an important prerequisite to ensuring a reliable and timely turnaround as well as to provide practical analytical tools for student training. RESULTS: We have developed an easy-to-use, interactive software tool for the analysis of multiple data sets comprising replicates of discrete bivariate data points. For each dataset, the software identifies the replicate data points from a defined matrix layout and calculates their means and standard errors. The averaged values are then automatically fitted using either a linear or a logistic dose response function. CONCLUSIONS: DRfit is a practical and convenient tool for the analysis of one or multiple sets of discrete data points acquired as replicates from multi-well plate assays. The design of the graphical user interface and the built-in analysis features make it a flexible and useful tool for a wide range of different assays.

Development of an HPLC-based guanosine monophosphate kinase assay and application to Plasmodium vivax guanylate kinase.

The development of a high-performance liquid chromatography (HPLC)-based method, for guanosine monophosphate kinase activity assays, is presented. The method uses the intrinsic UV absorption (at 260 nm) of substrates and products of the enzymatic reaction (GMP, ATP, ADP and GDP) to unambiguously determine percent conversion of substrate into product. It uses a commercially available C18 column which can separate reaction samples by elution under isocratic conditions in 12 min per run. The kinetics of the forward reaction catalyzed by Plasmodium vivax guanylate kinase (PvGK), a potential drug target against malaria, was determined. The relative concentrations of the two substrates (GMP and ATP) have a distinct effect on reaction velocity. Kinetic analyses showed the PvGK-catalyzed reaction to be associated with atypical kinetics, where substrate inhibition kinetics and non-Michaelis-Menten (sigmoidal) kinetics were found with respect to GMP and ATP, respectively. Additionally, the method was used in inhibition assays to screen twenty fragment-like compounds. The assays were robust and reproducible, with a signal window of 3.8 and a Z' factor of 0.6. For the best inhibitor, an IC50 curve was generated.

A convenient tool for bivariate data analysis and bar graph plotting with R.

The Java software jBar consists of a graphical user interface that allows the user to customize and assemble an included script for R. The scripted R pipeline calculates means and standard errors/deviations for replicates of numerical bivariate data and generates presentations in the form of bar graphs. A two-sided Student's t test is carried out against a user-selected reference and p-values are calculated. The user can enter the data conveniently through the built-in spreadsheet and configure the R pipeline in the graphical user interface. The configured R script is written into a file and then executed. Bar graphs can be generated as static PNG, PDF, and SVG files or as interactive HTML widgets. © 2019 International Union of Biochemistry and Molecular Biology, 47(2): 207-210, 2019.

Low Serum Albumin Levels are Associated with Increased 30-Day Cardiopulmonary Complications, Reoperation, and Readmission Rates Following Total Shoulder Arthroplasty.

Background: Hypoalbuminemia has been associated with several medical complications following surgery in a variety of orthopedic procedures. Hypoalbuminemia has previously been shown to have an increased risk for transfusions, hospital stay longer than three days, and mortality following total shoulder arthroplasty (TSA). This study seeks to further assess the relationship between low serum albumin and morbidity to allow surgeons to both preoperatively optimize patients and assess the risk of surgery prior to TSA. Methods: The American College of Surgeons National Surgical Quality Improvement Program® database was queried to identify 14,494 TSA patients, 6,129 (42.23%) who met inclusion criteria. Patients who had shoulder hemiarthroplasty, revision TSA, or incomplete serum albumin data were excluded. Demographic factors, preoperative comorbidities, and acute complication rates were assessed between hypoalbuminemic (n=485; 7.91%) and a propensity-matched control cohort (n=485), controlling for differences in patient demographics and comorbidities. Multivariate propensity-adjusted logistic regression analyses were used to assess hypoalbuminemia as an independent risk factor for specific postoperative complications. Results: Hypoalbuminemic patients undergoing TSA demonstrated significantly higher rates of pulmonary complications (p=0.006), unplanned intubation (p=0.014), DVT/PE (p=0.014), cardiac complications (p=0.033), infectious complications (p=0.025), blood transfusions (p<0.001), reoperation (p=0.007), extended length of stay (> 4 days) (p=0.036), unplanned readmission (p=0.001), and mortality (p=0.025) in the 30-day postoperative period when compared to the propensity-matched control cohort. On multivariate regression analyses, hypoalbuminemia independently increased the risk for pulmonary complications (OR 9.678, p=0.031), blood transfusions (OR 2.539, p<0.001), reoperation (OR 5.461, p=0.032), and readmission (OR 2.607, p=0.007). Conclusions: Hypoalbuminemic patients undergoing TSA had increased rates of overall cardiac and pulmonary complications, unplanned intubations, DVT/PE's, overall infectious complications, increased incidence of blood transfusions, reoperation, extended LOS (> 4 days), readmission, and death. Multivariate analyses demonstrated that low albumin was independently associated with increased risk for pulmonary complications, blood transfusions, reoperation, and readmission. Preoperative albumin levels in patients undergoing TSA may help with preoperative risk stratification and optimization.Level of evidence: III.

Validation of Cardiorespiratory Fitness Measurements in Adolescents.

Cardiorespiratory fitness (CRF) is an important indicator of adolescent cardiovascular well-being and future cardiometabolic health but not always feasible to measure. The purpose of this study was to estimate the concurrent validity of the non-exercise test (NET) for adolescents against the Progressive Aerobic Capacity Endurance Run (PACER®) and direct measures of VO2max as well as to examine the concurrent validity of the PACER® with a portable metabolic system (K4b2™). Forty-six adolescents (12-17 years) completed the NET prior to performing the PACER® while wearing the K4b2™. The obtained VO2max values were compared using linear regression, intra-class correlation (ICC), and Bland-Altman plots, and α was set at 0.05. The VO2max acquired directly from the K4b2™ was significantly correlated to the VO2max indirectly estimated from the NET (r = 0.73, p < 0.001, r2 = 0.53, ICC = 0.67). PACER® results were significantly related to the VO2max estimates from the NET (r = 0.81, p < 0.001, r2 = 0.65, ICC = 0.72). Direct measures from the K4b2™ were significantly correlated to the VO2max estimates from the PACER® (r = 0.87, p < 0.001, r2 = 0.75, ICC = 0.93). The NET is a valid measure of CRF in adolescents and can be used when an exercise test is not feasible.

Bovine Tuberculosis Management in Northwest Minnesota and Implications of the Risk Information Seeking and Processing (RISP) Model for Wildlife Disease Management.

Bovine tuberculosis (bTB) is an infectious, zoonotic disease caused by Mycobacterium bovis that can spread between domestic and wild animals, as well as to humans. The disease is characterized by the progressive development of lesions that compromise the victim's lungs and lymph system. The disease was first identified in northwest Minnesota in both cattle and white-tailed deer (Odocoileus virginianus) in 2005. Due to its risks to human and animal health, bTB has numerous implications related to population management, policy outcomes, stakeholder relations, and economic impacts. When dealing with complicated risks, like bTB, individuals often seek out and process information as a method to learn about, and cope, with the risk. We developed a questionnaire that adapted components of the Risk Information Seeking and Processing (RISP) model and surveyed northwest Minnesota deer hunters. Our objectives were to better understand how stakeholders perceive and act on information regarding disease management in wildlife and to understand the utility of the RISP model for such management contexts. We drew a random proportional sample of licensed deer hunters (n = 2100) from the area affected by bTB and conducted a multi-contact mail survey. We found that 43% of the variability in the information-seeking behaviors of respondents was explained by demographics, hunting importance, personal risk perceptions, attitudes, and subjective norms. However, these results are largely attributable to the factors in the RISP model encompassed by components of the Theory of Planned Behavior (i.e., attitudes, subjective norms, perceived behavioral control, and behavioral intentions). This information can help managers contextualize individuals' perceived risks to better frame communication efforts to address stakeholder concerns and develop best practices for disease communication. While the state of Minnesota is currently considered free of bTB, future outbreaks remain possible in Minnesota and elsewhere. Understanding the key factors in the processes through which deer hunters seek out information pertaining to the disease can help managers collect the data necessary to aid decisions about desired future management outcomes. In addition, testing RISP model performance in applied research improves its future use across a broad spectrum of topics throughout veterinary disease management.

Trehalose 6-phosphate phosphatases of Pseudomonas aeruginosa.

The opportunistic bacterium Pseudomonas aeruginosa has been recognized as an important pathogen of clinical relevance and is a leading cause of hospital-acquired infections. The presence of a glycolytic enzyme in Pseudomonas, which is known to be inhibited by trehalose 6-phosphate (T6P) in other organisms, suggests that these bacteria may be vulnerable to the detrimental effects of intracellular T6P accumulation. In the present study, we explored the structural and functional properties of trehalose 6-phosphate phosphatase (TPP) in P. aeruginosa in support of future target-based drug discovery. A survey of genomes revealed the existence of 2 TPP genes with either chromosomal or extrachromosomal location. Both TPPs were produced as recombinant proteins, and characterization of their enzymatic properties confirmed specific, magnesium-dependent catalytic hydrolysis of T6P. The 3-dimensional crystal structure of the chromosomal TPP revealed a protein dimer arising through ß-sheet expansion of the individual monomers, which possess the overall fold of halo-acid dehydrogenases.-Cross, M., Biberacher, S., Park, S.-Y., Rajan, S., Korhonen, P., Gasser, R. B., Kim, J.-S., Coster, M. J., Hofmann, A. Trehalose 6-phosphate phosphatases of Pseudomonas aeruginosa.

Enzyme characteristics of pathogen-specific trehalose-6-phosphate phosphatases.

Owing to the key role of trehalose in pathogenic organisms, there has recently been growing interest in trehalose metabolism for therapeutic purposes. Trehalose-6-phosphate phosphatase (TPP) is a pivotal enzyme in the most prominent biosynthesis pathway (OtsAB). Here, we compare the enzyme characteristics of recombinant TPPs from five important nematode and bacterial pathogens, including three novel members of this protein family. Analysis of the kinetics of trehalose-6-phosphate hydrolysis reveals that all five enzymes display a burst-like kinetic behaviour which is characterised by a decrease of the enzymatic rate after the pre-steady state. The observed super-stoichiometric burst amplitudes can be explained by multiple global conformational changes in members of this enzyme family during substrate processing. In the search for specific TPP inhibitors, the trapping of the complex conformational transitions in TPPs during the catalytic cycle may present a worthwhile strategy to explore.

Trehalose-6-phosphate phosphatase as a broad-spectrum therapeutic target against eukaryotic and prokaryotic pathogens.

As opposed to organism-based drug screening approaches, protein-based strategies have the distinct advantage of providing insights into the molecular mechanisms of chemical effectors and thus afford a precise targeting. Capitalising on the increasing number of genome and transcriptome datasets, novel targets in pathogens for therapeutic intervention can be identified in a more rational manner when compared with conventional organism-based methodologies. Trehalose-6-phosphate phosphatases (TPPs) are structurally and functionally conserved enzymes of the trehalose biosynthesis pathway which play a critical role for pathogen survival, in particular, in parasites. The absence of these enzymes and trehalose biosynthesis from mammalian hosts has recently given rise to increasing interest in TPPs as novel therapeutic targets for drugs and vaccines. Here, we summarise some key aspects of the current state of research towards novel therapeutics targeting, in particular, nematode TPPs.

Probing function and structure of trehalose-6-phosphate phosphatases from pathogenic organisms suggests distinct molecular groupings.

The trehalose biosynthetic pathway is of great interest for the development of novel therapeutics because trehalose is an essential disaccharide in many pathogens but is neither required nor synthesized in mammalian hosts. As such, trehalose-6-phosphate phosphatase (TPP), a key enzyme in trehalose biosynthesis, is likely an attractive target for novel chemotherapeutics. Based on a survey of genomes from a panel of parasitic nematodes and bacterial organisms and by way of a structure-based amino acid sequence alignment, we derive the topological structure of monoenzyme TPPs and classify them into 3 groups. Comparison of the functional roles of amino acid residues located in the active site for TPPs belonging to different groups reveal nuanced variations. Because current literature on this enzyme family shows a tendency to infer functional roles for individual amino acid residues, we investigated the roles of the strictly conserved aspartate tetrad in TPPs of the nematode Brugia malayi by using a conservative mutation approach. In contrast to aspartate-213, the residue inferred to carry out the nucleophilic attack on the substrate, we found that aspartate-215 and aspartate-428 of BmTPP are involved in the chemistry steps of enzymatic hydrolysis of the substrate. Therefore, we suggest that homology-based inference of functionally important amino acids by sequence comparison for monoenzyme TPPs should only be carried out for each of the 3 groups.-Cross, M., Lepage, R., Rajan, S., Biberacher, S., Young, N. D., Kim, B.-N., Coster, M. J., Gasser, R. B., Kim, J.-S., Hofmann, A. Probing function and structure of trehalose-6-phosphate phosphatases from pathogenic organisms suggests distinct molecular groupings.

Mutant p97 exhibits species-specific changes of its ATPase activity and compromises the UBXD9-mediated monomerisation of p97 hexamers.

p97 (VCP) is a homo-hexameric triple-A ATPase that exerts a plethora of cellular processes. Heterozygous missense mutations of p97 cause at least five human neurodegenerative disorders. However, the specific molecular consequences of p97 mutations are hitherto widely unknown. Our in silico structural models of human and Dictyostelium p97 showed that the disease-causing human R93C, R155H, and R155C as well as Dictyostelium R154C, E219K, R154C/E219K p97 mutations constitute variations in surface-exposed locations. In-gel ATPase activity measurements of p97 monomers and hexamers revealed significant mutation- and species-specific differences. While all human p97 mutations led to an increase in ATPase activity, no changes could be detected for the Dictyostelium R154C mutant, which is orthologous to human R155C. The E219K mutation led to an almost complete loss of activity, which was partially recuperated in the R154C/E219K double-mutant indicating p97 inter-domain communication. By means of co-immunoprecipitation experiments we identified an UBX-domain containing Dictyostelium protein as a novel p97 interaction partner. We categorized all UBX-domain containing Dictyostelium proteins and named the interaction partner UBXD9. Pull-down assays and surface plasmon resonance analyses of Dictyostelium UBXD9 or the human orthologue TUG/ASPL/UBXD9 demonstrated direct interactions with p97 as well as species-, mutation- and ATP-dependent differences in the binding affinities. Sucrose density gradient assays revealed that both human and Dictyostelium UBXD9 proteins very efficiently disassembled wild-type, but to a lesser extent mutant p97 hexamers into monomers. Our results are consistent with a scenario in which p97 point mutations lead to differences in enzymatic activities and molecular interactions, which in the long-term result in a late-onset and progressive multisystem disease.

Exploring the molecular mechanisms of parasite-host interactions with a view towards new therapeutics and vaccines.

Despite the massive disease burden worldwide caused by parasitic nematodes and other infectious pathogens, the molecular basis of many infectious diseases caused by these pathogens has been unduly neglected for a long time. Therefore, accelerated progress towards novel therapeutics, and ultimately control of such infectious diseases, is of crucial importance. Capitalising on the wealth of data becoming available from proteomic and genomic studies, new protein targets at the pathogen-host interface can be identified and subjected to protein-based explorations of the molecular basis of pathogen-host interactions. By combining the use of systems and structural biology methodologies, insights into the structural and molecular mechanisms of these interactions can assist in the development of therapeutics and/or vaccines. This brief review examines two different proteins from the body wall of blood flukes - annexins and the stress-induced phosphoprotein 1 - both of which are presently interesting targets for the development of therapeutics.

Panel docking of small-molecule libraries - Prospects to improve efficiency of lead compound discovery.

Computational docking as a means to prioritise small molecules in drug discovery projects remains a highly popular in silico screening approach. Contemporary docking approaches without experimental parametrisation can reliably differentiate active and inactive chemotypes in a protein binding site, but the absence of a correlation between the score of a predicted binding pose and the biological activity of the molecule presents a clear limitation. Several novel or improved computational approaches have been developed in the recent past to aid in screening and profiling of small-molecule ligands for drug discovery, but also more broadly in developing conceptual relationships between different protein targets by chemical probing. Among those new methodologies is a strategy known as inverse virtual screening, which involves the docking of a compound into different protein structures. In the present article, we review the different computational screening methodologies that employ docking of atomic models, and, by means of a case study, present an approach that expands the inverse virtual screening concept. By computationally screening a reasonably sized library of 1235 compounds against a panel of 48 mostly human kinases, we have been able to identify five groups of putative lead compounds with substantial diversity when compared to each other. One representative of each of the five groups was synthesised, and tested in kinase inhibition assays, yielding two compounds with micro-molar inhibition in five human kinases. This highly economic and cost-effective methodology holds great promise for drug discovery projects, especially in cases where a group of target proteins share high structural similarity in their binding sites.