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Our knowledge of the binding sites for neutralizing Abs (NAb) that recognize a broad range of HIV-1 strains (bNAb) has substantially increased in recent years. However, gaps remain in our understanding of how to focus B cell responses to vulnerable conserved sites within the HIV-1 envelope glycoprotein (Env). In this article, we report an immunization strategy composed of a trivalent HIV-1 (clade B envs) DNA prime, followed by a SIVmac239 gp140 Env protein boost that aimed to focus the immune response to structurally conserved parts of the HIV-1 and simian immunodeficiency virus (SIV) Envs. Heterologous NAb titers, primarily to tier 1 HIV-1 isolates, elicited during the trivalent HIV-1 env prime, were significantly increased by the SIVmac239 gp140 protein boost in rabbits. Epitope mapping of Ab-binding reactivity revealed preferential recognition of the C1, C2, V2, V3, and V5 regions. These results provide a proof of concept that a distally related retroviral SIV Env protein boost can increase pre-existing NAb responses against HIV-1.

The study of glucosinolates and their regulation has provided a powerful framework for the exploration of fundamental questions about the function, evolution, and ecological significance of plant natural products, but uncertainties about their metabolism remain. Previous work has identified one thiohydroximate S-glucosyltransferase, UGT74B1, with an important role in the core pathway, but also made clear that this enzyme functions redundantly and cannot be the sole UDP-glucose dependent glucosyltransferase (UGT) in glucosinolate synthesis. Here, we present the results of a nearly comprehensive in vitro activity screen of recombinant Arabidopsis Family 1 UGTs, which implicate other members of the UGT74 clade as candidate glucosinolate biosynthetic enzymes. Systematic genetic analysis of this clade indicates that UGT74C1 plays a special role in the synthesis of aliphatic glucosinolates, a conclusion strongly supported by phylogenetic and gene expression analyses. Finally, the ability of UGT74C1 to complement phenotypes and chemotypes of the ugt74b1-2 knockout mutant and to express thiohydroximate UGT activity in planta provides conclusive evidence for UGT74C1 being an accessory enzyme in glucosinolate biosynthesis with a potential function during plant adaptation to environmental challenge.

Aseptic failure of massive endoprostheses used in the reconstruction of major skeletal defects remains a major clinical problem. Fixation using compressive osseointegration was developed as an alternative to cemented and traditional press-fit fixation in an effort to decrease aseptic failure rates.The purpose of this study was to answer the following questions: (1) What is the survivorship of this technique at minimum 2-year followup? (2) Were patient demographic variables (age, sex) or anatomic location associated with implant failure? (3) Were there any prosthesis-related variables (eg, spindle size) associated with failure? (4) Was there a discernible learning curve associated with the use of the new device as defined by a difference in failure rate early in the series versus later on?The first 50 cases using compressive osseointegration fixation from two tertiary referral centers were retrospectively studied. Rates of component removal for any reason and for aseptic failure were calculated. Demographic, surgical, and oncologic factors were analyzed using regression analysis to assess for association with implant failure. Minimum followup was 2 years with a mean of 66 months. Median age at the time of surgery was 14.5 years.A total of 15 (30%) implants were removed for any reason. Of these revisions, seven (14%) were the result of aseptic failure. Five of the seven aseptic failures occurred at less than 1 year (average, 8.3 months), and none occurred beyond 17 months. With the limited numbers available, no demographic, surgical, or prosthesis-related factors correlated with failure.Most aseptic failures of compressive osseointegration occurred early. Longer followup is needed to determine if this technique is superior to other forms of fixation.

The aims of this study were to measure the accuracy of computerised tomography (CT) and magnetic resonance imaging (MRI) scans in detecting colorectal liver metastases (CRLM) and to determine if patients who are staged with MRI in addition to CT have longer liver recurrence-free survival compared to those having CT alone in a unit performing routine intra-operative ultrasound. A retrospective analysis of patients undergoing liver resection for CRLM was performed. Patients staged pre-operatively with CT or with additional MRI were included and those with additional PET imaging were excluded from survival analysis. Timing and site of tumour recurrence were recorded. During a 7-year period, 303 patients underwent resection for CRLM of whom 47 (15.5 %) were staged with CT alone and 36 (11.9 %) with additional MRI. The overall accuracy of CT (63 %) and MRI (61.9 %) was similar in the detection of tumour nodules (P = 0.905). There was no difference in the rate of intra-hepatic recurrence between groups with 13/47 and 8/36 cases, respectively (P = 0.737). There was no difference in the disease-free survival curves between the groups (P = 0.487). Our recommendation is that MRI should not be a mandatory imaging modality in referral guidelines for patients with hepatic CRLM, as the cost and delay associated with the scan outweigh any potential benefit in terms of improved sensitivity compared to CT.

In January, 2018, Fornal and Grinstein proposed that a previously unobserved neutron decay branch to a dark matter particle (χ) could account for the discrepancy in the neutron lifetime observed in two different types of experiments. One of the possible final states discussed includes a single χ along with an e+e− pair. We use data from the UCNA (Ultracold Neutron Asymmetry) experiment to set limits on this decay channel. Coincident electron-like events are detected with ∼ 4π acceptance using a pair of detectors that observe a volume of stored Ultracold Neutrons (UCNs). We use the timing information of coincidence events to select candidate dark sector particle decays by applying a timing calibration and selecting events within a physically-forbidden timing region for conventional n → p + e- + ν̅e decays. The summed kinetic energy (Ee+e−) from such events is reconstructed and used to set limits, as a function of the χ mass, on the branching fraction for this decay channel.

Introduction. The aim of this study was to assess the interaction of liver and renal dysfunction as risk factors for mortality after liver resection. Materials and Methods. A retrospective analysis of 501 patients undergoing liver resection in a single unit was undertaken. Posthepatectomy liver failure (PHLF) was defined according to the International Study Group of Liver Surgery (ISGLS) definition (assessed on day 5) and renal dysfunction according to RIFLE criteria. 90-day mortality was recorded. Results. Twenty-three patients died within 90 days of surgery (4.6%). The lowest mortality occurred in patients without evidence of PHLF or renal dysfunction (2.7%). The mortality rate in patients with isolated PHLF or renal dysfunction was 20% compared to 45% in patients with both. Diabetes (), renal dysfunction (), and PHLF on day 5 () were independent predictors of 90-day mortality. Discussion. PHLF and postoperative renal dysfunction are independent predictors of 90-day mortality following liver resection but the predictive value for mortality is significantly higher when failure of both organ systems occurs simultaneously.

Introduction: The aim of this study was to analyse the influence of factors reported in the minimum histopathology dataset for colorectal liver metastases (CRLM) and other pre-operative factors compared with additional data relating to the presence of tumour pseudocapsules and necrosis on recurrence 1 year after a resection. Methods: For a period of 14 months, extended histological reporting of CRLM specimens was performed, including the presence of pseudocapsules and necrosis in each tumour. The details of recurrence were obtained from surveillance imaging. Results: In 66 patients there were 27 recurrences within 1 year. The rates were lower for patients with tumour pseudocapsules (8/27) than for patients without (19/36) (P = 0.030). Pseudocapsules were associated with a younger age (P = 0.005), nodal stage of the primary colorectal tumour (P = 0.025) and metachronous tumours (P = 0.004). In patients with synchronous disease and pseudocapsules, the recurrence rate was 2/12 compared with 13/23 patients without pseudocapsules (P = 0.026). Discussion: These findings demonstrate that histological examination of resection specimens can provide significant additional prognostic information for patients after resection of CRLM, compared with clinical and radiological data. The present finding that the absence of a pseudocapsule in patients with synchronous CRLM is associated with a dramatically worse outcome may help direct patient-specific adjuvant treatment and care.

HIV-1 DNA vaccines have many advantageous features. Evaluation of HIV-1 vaccine candidates often starts in small animal models before macaque and human trials. Here, we selected and optimized DNA vaccine candidates through systematic testing in rabbits for the induction of broadly neutralizing antibodies (bNAb). We compared three different animal models: guinea pigs, rabbits and cynomolgus macaques. Envelope genes from the prototype isolate HIV-1 Bx08 and two elite neutralizers were included. Codon-optimized genes, encoded secreted gp140 or membrane bound gp150, were modified for expression of stabilized soluble trimer gene products, and delivered individually or mixed. Specific IgG after repeated i.d. inoculations with electroporation confirmed in vivo expression and immunogenicity. Evaluations of rabbits and guinea pigs displayed similar results. The superior DNA construct in rabbits was a trivalent mix of non-modified codon-optimized gp140 envelope genes. Despite NAb responses with some potency and breadth in guinea pigs and rabbits, the DNA vaccinated macaques displayed less bNAb activity. It was concluded that a trivalent mix of non-modified gp140 genes from rationally selected clinical isolates was, in this study, the best option to induce high and broad NAb in the rabbit model, but this optimization does not directly translate into similar responses in cynomolgus macaques.

Background: Plant natural products typically contain regiospecific modifications in their side chains that lead to different bioactivities. Results: Only 3-O-methylated flavonols enhance IL-1β production in THP-1 cells costimulated with the Toll-like receptor 2 agonist Pam3CSK4. Conclusion: Regiospecific methylation of flavonols controls their bioactivity as immunomodulators. Significance: This study provides a platform to explore the use of regiospecific-modified natural products as novel immunomodulators., It is now recognized that innate immunity to intestinal microflora plays a significant role in mediating immune health, and modulation of microbial sensing may underpin the impact of plant natural products in the diet or when used as nutraceuticals. In this context, we have examined five classes of plant-derived flavonoids (flavonols, flavones, flavanones, catechins, and cyanidin) for their ability to regulate cytokine release induced by the Toll-like receptor 2 (TLR2) agonist Pam3CSK4. We found that the flavonols selectively co-stimulated IL-1β secretion but had no impact on the secretion of IL-6. Importantly, this costimulation of TLR2-induced cytokine secretion was dependent on regiospecific methylation of the flavonol scaffold with a rank order of quercetin-3,4′-dimethylether > quercetin-3-methylether > casticin. The mechanism underpinning this costimulation did not involve enhanced inflammasome activation. In contrast, the methylated flavonols enhanced IL-1β gene expression through transcriptional regulation, involving mechanisms that operate downstream of the initial NF-κB and STAT1 activation events. These studies demonstrate an exquisite level of control of scaffold bioactivity by regiospecific methylation, with important implications for understanding how natural products affect innate immunity and for their development as novel immunomodulators for clinical use.

To evaluate the clinical results of open reduction and internal fixation (ORIF) in proximal humerus fractures with delayed presentation. Retrospective comparative study. Level III. From June 2005 to June 2010, thirty-nine (39) proximal humerus fractures were treated with ORIF after a delay of 21–120 days from the initial injury. Patients were divided into three groups: (1) isolated 2-part greater tuberosity fractures; (2) part surgical neck fractures; and (3) 3- and 4-part complex fractures. The range of motion (ROM), visual analogue scale (VAS), Constant–Murley score, University of California Los Angeles (UCLA) scoring system score and Simple Shoulder Test (SST) score were all recorded. The results were analyzed with the use of the Mann–Whitney U test and stratified by age, gender, side of injury, interval from injury to surgery, and postoperative functional results. Additionally, the results of different fracture types and complications were compared across the three groups. The mean forward flexion was 143.8° ± 28.9°, external rotation was 33.2° ± 19.6°, and internal rotation was up to the T10 level. The mean VAS was 0.8 ± 1.2; the mean Constant score was 82.0 ± 15.0; the UCLA score was 27.2 ± 7.1; and the mean SST was 9.5 ± 2.0 at the last follow-up. There were no significant differences among the three groups except in internal rotation. Compared to those without any complications, patients with complications demonstrated worse ROM and a lower functional score (p

Artemisia annua L. is an annual shrub in the Asteraceae that is commercially grown and cropped in Southeast Asia, Africa, Madagascar and India for the antimalarial compound, artemisinin (ART). Demand for a stable supply chain to artemisinin combination therapies has driven forward complementary programs of crop improvement and microbially based semisynthesis to boost global ART production. Recently, photochemical flow-reactor technology has been developed and proposed as an additional input to the supply chain by converting either artemisinic acid (AA) or dihydroartemisinic acid (DHAA) to ART. These compounds are naturally present in A. annua but their content in available commercial lines is unclear. Since 2006 we have undertaken a program to improve A. annua ART yields using diverse germplasm and modern molecular breeding tools. Sesquiterpenoid extracts from eleven diverse populations with low to high ART contents grown under glasshouse conditions had population median AA or DHAA contents never exceeding 0.9% of leaf dry weight (DW), but ART achieved a maximum median value of 1.5% DW. We also field-trialed 280 distinct lines selected for high ART content including controls, locally grown commercial lines and test hybrids in the major global growing regions over 2010–2011. The trend was for ART to exceed AA or DHAA and content never exceeded 1.6% DW for any compound. These results indicate that currently grown A. annua lines are poor sources of AA and DHAA relative to ART. However, a glasshouse-based forward-screen of over 14,000 individuals from a mutagenized population suggested that increased DHAA yields may be achievable.

Data from the Sudbury Neutrino Observatory have been used to constrain the lifetime for nucleon decay to ``invisible'' modes, such as n -> 3 nu. The analysis was based on a search for gamma-rays from the de-excitation of the residual nucleus that would result from the disappearance of either a proton or neutron from O16. A limit of tau_inv > 2 x 10^{29} years is obtained at 90% confidence for either neutron or proton decay modes. This is about an order of magnitude more stringent than previous constraints on invisible proton decay modes and 400 times more stringent than similar neutron modes., Update includes missing efficiency factor (limits change by factor of 2) Submitted to Physical Review Letters

Remote condition monitoring and validation of railway points were discussed. Recent developments in microelectronics and information technology (IT) made possible low-cost, high-quality condition monitoring for trackside equipment. These equipments reduced the likelihood to breakdown and to enhance railway service quality. The results showed that there was value in continuous monitoring of the overall infrastructure which provided fundamentally new data not available under existing maintenance practice.

A new implementation of a Coriolis mass-flow meter transmitter is described. It is based on digital components, and has improved performance compared with the commercial, mostly analogue, transmitter using the same flowtube (transducer). Improvements are found in flowtube control, measurement precision, and performance with two-phase and partially-empty conditions, including batching from empty. The new transmitter is viewed as a second-generation sensor validation (SEVA) demonstrator, in which experience from validating the commercial analogue transmitter has led to a redesign using digital technology. The resulting SEVA transmitter provides improved measurement performance and reduced vulnerability to fault conditions, as well as on-line estimates of measurement quality and fault compensation (Henry and Clarke, Control Engineering practice, 1 (4) (1993) 585-610). (C) 2000 Elsevier Science Ltd. All rights reserved.

Misdiagnosis of heparin-induced thrombocytopenia (HIT) is common and exposes patients to high-risk therapies and potentially serious adverse events. The primary objective of this study was to evaluate the impact of collaboration between an inpatient pharmacy-driven anticoagulation management service (AMS) and hospital reference laboratory to reduce inappropriate HIT antibody testing via pharmacist intervention and use of the 4T pre-test probability score. Secondary objectives included clinical outcomes and cost-savings realized through reduced laboratory testing and decreased unnecessary treatment of HIT. This was a single center, pre-post, observational study. The hospital reference laboratory contacted the AMS when they received a blood sample for an enzyme-linked immunosorbent HIT antibody (HIT Ab). Trained pharmacists prospectively scored each HIT Ab ordered by using the 4T score with subsequent communication to physicians recommending for or against processing and reporting of lab results. Utilizing retrospective chart review and a database for all patients with a HIT Ab ordered during the study period, we compared the incidence of HIT Ab testing before and after implementation of the pharmacy-driven 4T score intervention. Our intervention significantly reduced the number of inappropriate HIT Ab tests processed (176 vs. 63, p

Antimicrobial resistance presents a threat to quality patient care. Knowledge of localantibacterial susceptibility patterns can guide clinicians in empiric antibacterial administration andassist pharmacists and infectious disease physicians in development of appropriate therapeutic pathways.To characterize Wisconsin antibacterial susceptibility patterns and elucidate geographicor temporal variation in antibacterial resistance, a retrospective, observational analysis of antibiogram data was performed. Seventy-two members of the Wisconsin Clinical Laboratory Network(WCLN) submitted antibiograms describing clinically significant isolates tested in calendar year 2013 to the WCLN Laboratory Technical Advisory Group.In the context of commonly reported antibacterial agents, data were compiled for approximately 75,800 isolates of Escherichia coi; 13,300 Klebsiella pneumoniae; 6300 Proteus mirobilis;2800 Enterobacter cloacae; 8400 Pseudomonas aeruginosa; 30,000 S aureus; 11,200 coagulase-negative Staphylococcus spp; and 13,800 Enterococcus spp. P mirobilis isolates from northern Wisconsin were more likely to demonstrate resistance than those in the southern region. In contrast, P aeruginosa isolates from southern Wisconsin had decreased susceptibility to a number ofagents when compared to other regions. Temporal trending in decreased E coli and P mirabilis susceptibility to fluoroquinolones and trimethoprimsulfamethoxazole was observed. Increased methicillin-resistant Staphylococcus oureus (MRSA) rates were observed in northwest and southeastWisconsin. In general, northeast Wisconsin exhibited less frequency of antibacterial resistance.Geographic variation exists with respect to antibacterial resistance, particularly inareas of Wisconsin adjacent to large population centers of neighboring states. Antibacterial surveillance in Wisconsin is indicated on a regular basis to assess emerging trends in antibacterial resistance. Existing WCLN infrastructure allows for such investigations.

Background: Differentiating hepatic mucinous cystic neoplasms (MCNs) from simple hepatic cysts (SCs) preoperatively is a challenging task. Our aim was to determine whether radiological features on ultrasound scan (USS), CT or MRI, cyst fluid tumour markers, or multidisciplinary team (MDT) outcomes could differentiate MCN from SC. Methods: A retrospective review of radiological features, cyst fluid tumour marker levels and MDT outcomes in 52 patients was performed. Results: There were 13 patients with MCN, 38 with SC and one ciliated foregut cyst. MCNs were more often solitary (p = 0.006). Although no other individual radiological characteristic on USS, CT or MRI was predictive of MCN, MDT outcomes stating that a cyst was complex in nature were highly predictive (p = 0.0007). Cyst fluid carbohydrate antigen 19-9, carcino-embryonic antigen and cancer antigen 125 were unable to differentiate MCN from SC (p = 0.45, p = 0.49, and p = 0.73, respectively). Conclusions: MDT outcomes are of greatest value when trying to differentiate MCN from SC, as well as having a solitary cyst on imaging. Conventional cyst fluid tumour markers are unhelpful. All suspicious cystic liver lesions should be discussed pre-operatively by a hepatobiliary MDT to determine the most appropriate surgical approach.

Despite the frequency of open fractures, their management remains one of the greatest and most debated orthopedic challenges. The current challenges that health care faces financially and clinically offer an opportunity to develop a universal reliable, reproducible, evidence-based protocol. The authors review the current evidence concerning the acute management of open fractures and suggest a modern treatment algorithm.

Purpose We compare autologous bone graft volume from three different harvest sites and the associated pain levels at one and four weeks post-harvest. Methods We prospectively collected a total of 47 consecutive autologous bone grafts in our level I trauma centre between July 2010 and September 2010 – 12 from the anterior iliac crest (AIC), 19 from the olecranon and 16 from the lateral aspect of the proximal tibia. Results The mean ages for each group were 44.8, 41.9 and 42.1, respectively, with no statistically significant differences for age or gender. The mean quantities harvested were, respectively, 6.2 cm3 (cubic centimetres) from the AIC, 5.7 cm3 from the olecranon and 7.3 cm3 from the proximal tibia, with a statistically superior quantity from the proximal tibia when compared to the olecranon and no differences between the other sites. Analysis of the visual analogue scale (VAS) at one week and four weeks post-harvest confirmed statistically that pain was more severe at the AIC site when compared to both the olecranon and proximal tibia sites. There were no infections or neurovascular complications. Conclusions We conclude that the proximal tibia and olecranon are acceptable alternatives to the anterior iliac crest for autologous bone graft and may be preferred when the location of the primary injury permits.

A simple, sensitive, and rapid chemiluminescence method was developed for the detection and quantification of selected derivatives of artemisinin. It was found that the chemiluminescence (CL) signal resulting from the alkaline luminol reaction of the artemisinin derivatives in the presence of hematin was linear over a wide concentration range. We report results that suggest that a visual test or a test involving digital image capture may be used in addition to a field-ready instrumental approach to combat drug counterfeiting of malarial therapies.

The potential effects of abstraction for water supply on salmonid populations in England and Wales are reviewed. The duties of water resource planning and the contribution of the water industry investment in river basin management planning are discussed. Given the possible effects of climate change on public water supply and demand, and the uncertainty of hydroecological relationships, key principles to ensure long-term investment by water utility companies to meet European Union Water Framework Directive standards achieve the best benefit for salmonids are recommended.

Analysis of longitudinally obtained HIV-1 env sequences from an individual with reported cross-reactive neutralizing activity revealed that the majority of viral variants obtained from serum between 4 and 7 years after seroconversion were unable to persist in peripheral blood. Here we show that these viral variants were more sensitive to autologous serum neutralization, had shorter envelopes with fewer potential N-linked glycosylation sites, and showed lower replication kinetics than successfully evolving HIV-1 variants. These data reflect the host selection pressures on phenotypic characteristics of HIV-1 and illustrate in detail the dynamic interaction between HIV-1 and its host's humoral immune responses.

We describe a simple drift tube counter that has been used as a cosmic ray veto for the UCNA experiment, a first-ever measurement of the neutron beta-asymmetry using ultra-cold neutrons. These detectors provide an inexpensive alternative to more conventional scintillation detectors for large area cosmic ray anticoincidence detectors.

The HIV-1 quasispecies in peripheral blood mononuclear cells (PBMC) is considered to be a mix of actively replicating, latent, and archived viruses and may be genetically distinct from HIV-1 variants in plasma that are considered to be recently produced. Here we analyzed the genetic relationship between gp160 env sequences from replication competent clonal HIV-1 variants that were isolated from PBMC and from contemporaneous HIV-1 RNA in serum and HIV-1 proviral DNA in PBMC of four longitudinally studied therapy naive HIV-1 infected individuals. Replication competent clonal HIV-1 variants, HIV-1 RNA from serum, and HIV-1 proviral DNA from PBMC formed a single virus population at most time points analyzed. However, an under-representation in serum of HIV-1 sequences with predicted CXCR4 usage was sometimes observed implying that the analysis of viral sequences from different sources may provide a more complete assessment of the viral quasispecies in peripheral blood in vivo. (C) 2010 Elsevier Inc. All rights reserved

Van Amburgh's methodology for allocating personal and enterprise goodwill is flawed. We present a standard empirical approach that can be applied to the problem of allocating goodwill. Based on survey data of professional accounting firms, we apply regression analysis and estimate the percentage of total goodwill that is personal goodwill. These results suggest that personal goodwill for relatively small professional accounting practices ranges from approximately 45% to 88% of total goodwill. The same methodology and data are also applied to the problem of determining an individual owner's share of total personal goodwill.

The Art of Artemisia As the malaria parasite, which is transmitted through mosquito vectors, develops resistance, previously useful control mechanisms are beginning to fail. Combination therapies based on the plant product artemisinin are a promising alternative. Graham et al. (p. 328 ; see the Perspective by Milhous and Weina ) have now developed a genetic map of the plant Artemisia annua from which artemisinin is derived. The results lay the foundation for improving agricultural productivity of this natural product, which is becoming increasingly important in the fight against malaria.

This study addresses the potential risk to the genetic resources of UK sheep breeds from endemism. Twelve native breeds are analysed that exist in significant numbers and continue to be commercially farmed livestock. The breeds were selected to represent regions of the British Isles: North of England hill breeds (Herdwick, Rough Fell, Lonk, Dalesbred), South of England breeds (Southdown, Romney, Devon Closewool, Exmoor Horn), Scottish breed (South Country Cheviot) and Welsh/Welsh border breeds (Welsh Hill Speckled Face, South Wales Mountain (Nelson), Clun Forest). For each breed, numerical data were collected in collaboration with breed society members and analysed in terms of population size and structure together with the extent of their geographical range. The number of flocks per breed proved to be highly variable, questioning the assumption that has been made that the number of breeding units can be disregarded in the calculations of endangerment. The data also indicated that an average flock size for a breed cannot be estimated to gain an insight of the population structure, since this parameter was found to vary considerably within and between breeds. The endemism was best illustrated in the breed maps which clearly highlighted the degree to which each breed was associated with a distinct geographical area. From the maps, ten of the twelve breeds analysed were concentrated, but the flocks of two breeds were visually more dispersed. In numerical terms, the ten breeds were found to have up to 95% of their breed numbers concentrated within a radius of 65 km from the mean centre of each breed. The study provides a valuable foundation for future research into genetic diversity within and between the sheep breeds analysed, a baseline of information to enable population trends to be examined and a robust evidence-base for policy decisions on Farm Animal Genetic Resources (FAnGR).

Summary The efficacy and safety of verapamil and nifedipine were objectively assessed in patients with chronic stable angina. Twenty four patients entered a double blind randomized cross over trial of nifedipine (10 mg thrice daily) and placebo. In this dosage nifedipine did not show any significant change in exercise duration and the variables obtained using computer assisted exercise testing when compared to placebo. The next stage consisted of another double blind randomized cross over trial comparing the effects of verapamil (120 mg thrice daily) and nifedipine (20 mg thrice daily) with an initial placebo run-in period in 32 patients. At this dose level nifedipine showed a definite and significant improvement in all the objective variables; however an increase in side effects was observed resulting in withdrawal of the drug in seven patients. A common problem was tachycardia precipitating angina after nifedipine ingestion. On the other hand verapamil produced a marked improvement in exercise tolerance and other variables as compared to placebo and nifedipine, was well tolerated and produced a mild bradycardia. This study clearly indicates that verapamil is distinctly superior to nifedipine in efficacy side effects and safety in patients with chronic stable angina. This may be attributable to the differential effect on heart rates induced by these drugs.

Plants, as predominantly sessile organisms, have evolved complex detoxification pathways to deal with a diverse range of toxic chemicals. The elasticity of this stress response system additionally enables them to tackle relatively recently produced, novel, synthetic pollutants. One such compound is the explosive 2,4,6-trinitrotoluene (TNT). Large areas of soil and groundwater are contaminated with TNT, which is both highly toxic and recalcitrant to degradation, and persists in the environment for decades. Although TNT is phytotoxic, plants are able to tolerate low levels of the compound. To identify the genes involved in this detoxification process, we used microarray analysis and then subsequently characterized seven uridine diphosphate (UDP) glycosyltransferases (UGTs) from Arabidopsis thaliana (Arabidopsis). Six of the recombinantly expressed UGTs conjugated the TNT-transformation products 2- and 4-hydroxylaminodinitrotoulene, exhibiting individual bias for either the 2- or the 4-isomer. For both 2- and 4-hydroxylaminodinitrotoulene substrates, two monoglucose conjugate products, confirmed by HPLC-MS-MS, were observed. Further analysis indicated that these were conjugated by either an O- or C-glucosidic bond. The other major compounds in TNT metabolism, aminodinitrotoluenes, were also conjugated by the UGTs, but to a lesser extent. These conjugates were also identified in extracts and media from Arabidopsis plants grown in liquid culture containing TNT. Overexpression of two of these UGTs, 743B4 and 73C1, in Arabidopsis resulted in increases in conjugate production, and enhanced root growth in 74B4 overexpression seedlings. Our results show that UGTs play an integral role in the biochemical mechanism of TNT detoxification by plants.

The synthesis of terpenoid glycosides typically uses a chemical strategy since few biocatalysts have been identified that recognise these scaffolds. In this study, a platform of 107 recombinant glycosyltransferases (GTs), comprising the multigene family of small molecule GTs of Arabidopsis thaliana have been screened against a range of model terpenoid acceptors to identify those enzymes with high activity. Twenty-seven GTs are shown to glycosylate a diversity of mono-, sesqui- and diterpenes, such as geraniol, perillyl alcohol, artemisinic acid and retinoic acid. Certain enzymes showing substantial sequence similarity recognise terpenoids containing a primary alcohol, irrespective of the linear or cyclical structure of the scaffold; other GTs glycosylate scaffolds containing secondary and tertiary alcohols; the carboxyl group of other terpenoids also represents a feature that is recognized by GTs previously known to form glucose esters with many different compounds. These data underpin the rapid prediction of potential biocatalysts from GT sequence information. To explore the potential of GTs as biocatalysts, their use for the production of terpenoid glycosides was investigated by using a microbial-based whole-cell biotransformation system capable of regenerating the cofactor, UDP-glucose. A high cell density fermentation system was shown to produce several hundred milligrams of a model terpenoid, geranyl-glucoside. The activities of the GTs are discussed in relation to their substrate recognition and their utility in biotransformations as a complement or alternative to chemical synthesis.

Plant Family 1 glycosyltransferases (GTs) recognize a wide range of natural and non-natural scaffolds and have considerable potential as biocatalysts for the synthesis of small molecule glycosides. Regiospecificity of glycosylation is an important property, given that many acceptors have multiple potential glycosylation sites. This study has used a domain-swapping approach to explore the determinants of regiospecific glycosylation of two GTs of Arabidopsis thaliana, UGT74F1 and UGT74F2. The flavonoid quercetin was used as a model acceptor, providing five potential sites for O-glycosylation by the two GTs. As is commonly found for many plant GTs, both of these enzymes produce distinct multiple glycosides of quercetin. A high performance liquid chromatography method has been established to perform detailed steady-state kinetic analyses of these concurrent reactions. These data show the influence of each parameter in determining a GT product formation profile toward quercetin. Interestingly, construction and kinetic analyses of a series of UGT74F1/F2 chimeras have revealed that mutating a single amino acid distal to the active site, Asn-142, can lead to the development of a new GT with a more constrained regiospecificity. This ability to form the 4 ′-O-glucoside of quercetin is transferable to other flavonoid scaffolds and provides a basis for preparative scale production of flavonoid 4 ′-O-glucosides through the use of whole-cell biocatalysis.

Plants produce p-aminobenzoate (pABA) in chloroplasts and use it for folate synthesis in mitochondria. In plant tissues, however, pABA is known to occur predominantly as its glucose ester (pABA-Glc), and the role of this metabolite in folate synthesis has not been defined. In this study, the UDP-glucose:pABA acyl-glucosyltransferase (pAGT) activity in Arabidopsis extracts was found to reside principally (95%) in one isoform with an apparent Km for pABA of 0.12 mm. Screening of recombinant Arabidopsis UDP-glycosyltransferases identified only three that recognized pABA. One of these (UGT75B1) exhibited a far higher kcat/Km value than the others and a far lower apparent Km for pABA (0.12 mm), suggesting its identity with the principal enzyme in vivo. Supporting this possibility, ablation of UGT75B1 reduced extractable pAGT activity by 95%, in vivo [14C]pABA glucosylation by 77%, and the endogenous pABA-Glc/pABA ratio by 9-fold. The Keq for the pABA esterification reaction was found to be 3 × 10-3. Taken with literature data on the cytosolic location of pAGT activity and on cytosolic UDP-glucose/UDP ratios, this Keq value allowed estimation that only 4% of cytosolic pABA is esterified. That pABA-Glc predominates in planta therefore implies that it is sequestered away from the cytosol and, consistent with this possibility, vacuoles isolated from [14C]pABA-fed pea leaves were estimated to contain≥88% of the [14C]pABA-Glc formed. In total, these data and the fact that isolated mitochondria did not take up [3H]pABA-Glc, suggest that the glucose ester represents a storage form of pABA that does not contribute directly to folate synthesis.

Bulk GaAs crystals were grown by synthesis solute diffusion (SSD) technique in a wide range of growth temperatures between 990 and 1150 °C. Electrical properties of these crystals were studied by means of van der Pauw, admittance spectroscopy, deep levels transient spectroscopy and photoinduced current spectroscopy techniques. It was shown that the main defects determining the properties were the GaAs antisites acceptors and the A center acceptors with the levels, respectively, Ev +0.078 eV and Ev +0.43 eV. The conductivity of the grown crystals was p-type and showed a pronounced maximum at a level of 104–105 Ω cm for growth temperatures between 1020 and 1080 °C. If the crystals were additionally compensated either by unintentional Si donors contamination from quartz crucibles or by intentional light Te doping one could get semi-insulating material with the room temperature resistivity higher than 106 Ω cm. The Fermi level in such crystals was pinned near Ec −0.8 eV, i.e. close to the EL2 donors. Measurements by deep levels transient spectroscopy on n-type doped crystals or by low frequency capacitance–voltage on semi-insulating crystals showed that the density of EL2 in these samples was in the low 1014 cm−3 and that thus the EL2 donors were not the main compensating agents.

A period of recovery is commonly allowed between completion of chemotherapy for colorectal liver metastases (CRLM) and resection, during which tumour progression may occur. The study-aim is to assess the growth of CRLM in this interval and association with outcome.Data on 146 patients were analysed. Change in tumour size was assessed by comparing size determined by imaging performed on completion of chemotherapy with that determined by examination of the resected specimen, categorised by RECIST criteria.In the interval before surgery sixteen patients (11%) fulfilled criteria for partial response (PR), 48 (33%) had stable disease (SD) and 82 (56%) had progressive disease (PD). Among patients with PD following chemotherapy the median disease-free survival of patients who initially responded (26 months) was longer than in those who initially had stable disease (7 months) (P = 0.002). No association was noted between rate of tumour growth after completion of chemotherapy and disease-free survival.Change in tumour size after completion of chemotherapy is variable and can be rapid, especially in patients who initially respond to treatment. However, disease-free survival is determined by tumour behaviour during treatment and not by change in size after completion of chemotherapy.

Background: Delay between diagnosis of peri-ampullary cancer (PC) and surgery may allow tumour progression and affect outcome. This study aims to explore associations of interval to surgery (IS) with pathological outcomes and survival in patients with PC. Method: A database review of all patients undergoing surgery between 2006 and 2014 was undertaken. IS was measured from diagnosis by imaging. Potential association between IS and survival was measured using Cox regression analysis, and between IS and pathological outcome with multivariate logistic analysis. Results: 388 patients underwent surgery. The median IS was 49 days (1-551 days), and was not associated with any of the evaluated outcomes in patients with pancreatic (149) or distal bile duct (46) cancer. For patients with ampullary cancer (71) longer IS was associated with improved survival, with median survival of 1.3, 3.1 and 4.3 years for patients waiting 2 (25) months for surgery (p=0.036). A higher rate of margin positivity (31.4%) was also noted among patients who waited less than the median IS compared to those waiting longer than this interval (11.4%) (p=0.032). Conclusion: For patients with ampullary cancer there is a paradoxical improvement in outcome among those with a longer IS, which may be explained by progression to inoperability of more aggressive lesions.

A radiochemical 71 Ga- 71 Ge experiment to determine the primary flux of neutrinos from the Sun began measurements of the solar neutrino flux at the Baksan Neutrino Observatory in 1990. The number of 71 Ge atoms extracted from 30 tons of gallium in 1990 and from 57 tons of gallium in 1991 was measured in 12 runs during the period of January 1990 to December 1991. The combined 1990 and 1991 data-sets give a value of 58 + 17/ —24 (stat) ± 14 (syst) SNU. This is to be compared with 132 + 7/ —5 SNU predicted by the Standard Solar Model.

The earlier search at HERMES for narrow baryon states excited in quasireal photoproduction, decaying through the channel pK(S)(0) -> p pi(+)pi(-), has been extended with improved decay-particle reconstruction, more advanced particle identification, and increased event samples. The structure observed earlier at an invariant mass of 1528 MeV shifts to 1522 MeV and the statistical significance drops to about 2 sigma for data taken with a deuterium target. The number of events above background is 68(-31)(+98)(stat) +/- 13(sys). No such structure is observed in the hydrogen data set.

BackgroundAseptic complications such as stress shielding leading to bone loss are major problems associated with revision of cemented and uncemented long-stem tumor endoprostheses. Endoprosthetic reconstruction using compressive osseointegration fixation is a relatively new limb salvage technology designed to enhance osseointegration, prevent stress shielding, and provide fixation for short end-segments.Questions/purposes(1) What is the survivorship of this technique at minimum 5-year followup? (2) Were patient factors (age, sex, body mass index), oncological factors, or anatomic locations associated with implant failure? (3) Were there any prosthesis-related variables associated with failure?MethodsA single-center, retrospective review of patients with a minimum 5-year followup (mean, 8 years; range, 5-12 years) treated with an osseointegration compressive device for endoprosthetic fixation of proximal and distal femoral limb salvage reconstructions was performed. We have previously published the implant survivorship of this patient cohort with a minimum 2-year followup and are now reporting on the 5-year survivorship data. From 2002 to 2008, we performed 22 such procedures in 22 patients. Four patients died of their disease within 5 years of surgery and all surviving patients (n = 18) had complete followup data at a minimum of 5 years. General indications for this device during that time were pediatric and adult patients requiring primary endoprosthetic reconstructions of the proximal or distal femur for benign and malignant bone lesions. The primary outcome was reoperations for mechanical (aseptic) failures. Secondary outcomes included implant removal for nonmechanical failures and any patient-, oncological-, or implant-related variables associated with implant removal.ResultsAt a minimum of 5 years followup, overall mechanical (aseptic) implant survivorship was 16 of 18. Survivorship for all modes of failure (oncological failure, infection, arthrofibrosis, and mechanical failure) was 12 of 18. All mechanical failures occurred early, within the first 30 months. We identified no patient-, oncological-, or implant-related features predictive of failure.ConclusionsOur intermediate-term experience with compressive osseointegration fixation for endoprosthetic limb reconstructions demonstrates with longer clinical followup, no additional mechanical failures were observed as compared with our early analysis. Our experience with this fixation at a minimum of 5-years followup adds to a very limited but increasing body of literature demonstrating that after a transient period of increased risk for implant failures, survivorship stabilizes. Assessment of this fixation strategy beyond 10 years of clinical followup is needed.Level of evidenceLevel IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence.

Most of the marine sediment cores collected in the past 10 years by the ODP and IMAGES programs have been processed using a multi-sensor core logger (MSCL, Geotek). This system estimates the density of sediment using the γ-ray attenuation of a 137Cs source. To date, optically stimulated luminescence (OSL) dating has not been applied to sediments from cores treated in this way, because of the possible impact of this exposure on the existing luminescence signal. We have investigated the effect of such γ-ray irradiation by test exposure, measurement and estimation, and conclude that the usual 137Cs exposure from γ-ray attenuation densitometry has no impact on the luminescence signal of quartz grains in sediment cores. For longer exposure to the 137Cs source, a formula is provided to calculate the received dose. This result will permit the OSL dating technique to be applied to the voluminous body of marine cores already collected and processed by MSCL.