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In recent decades, out-migration has become a key livelihood strategy for many rural households in the middle hills region of Nepal. In this region, rural communities are key actors in the management of local resources such as community forests. Analysis of the link between community forests and out-migration is largely missing in the literature, even though the demographic changes associated with out-migration affect forest resource use and management. This article discusses how rural communities and traditional landscapes in the middle hills are changing following out-migration and how this process has changed the management by rural communities of local forests (including community forests). Overall, the research found a reduced dependency by households on forest products (e.g. firewood, fodder and timber) sourced from community forests. Also, the reduced demand is being supplied increasingly from trees/forests grown on private farmland, including natural regenerated forests and trees planted on abandoned farmland. The declining need for forest products and the lack of an economic incentive for active forest management coupled with a decreasing sense of community has reduced the interest in community forests, leading to less intensive and infrequent forest management. Given the prevalence of out-migration and the changing socio-economic context in the middle hills, it appears time to reconsider the management of community forests beyond a narrow range of uses to enable greater commercialization and encourage ecosystem services to be harnessed so community forests better align with contemporary rural livelihoods and landscapes.

We offer a unified conceptual, behavioral, and econometric framework for optimal fundraising that deals with both synergies and discrepancies between approaches from Economics, Marketing, Psychology, and Sociology. The purpose is to offer a framework that can bridge differences and open a dialogue between disciplines in order to facilitate optimal fundraising design. The literature is extensive, and our purpose is to offer a brief background and perspective on each of the approaches, provide an integrated framework leading to new insights, and discuss areas of future research.

Environmental and development discourses in China can be categorised into three narrative motifs framing human–nature relationships: peasant, indigenous, and community. Indigenous and community narratives have been widely adopted by environmental NGOs (eNGOs) in China in promoting community-based natural resource management projects, but there has been very limited critical research on such phenomena. Analysis of socio-economic change in two ethnic minority communities in Yunnan shows that neither narrative theme is fully internalised by the relevant communities. Instead narratives may be strategically modified or even rejected by local communities. This is due to different agendas being held by local communities and eNGOs, and two factors pertinent to rural China: the incompatibility of concepts of ‘community’ in Chinese and international contexts results in confusion, and a lack of recent territorial and cultural claims by rural communities since the collectivist era makes it difficult to construct the identity of a community. It remains challenging for eNGOs in China to advocate either community or indigenous narratives in contexts of rapid socio-economic change.

Background In 1980, smallpox disease was eradicated from nature and Variola virus, the etiological agent of smallpox, was confined to two laboratories, one located in Russia (Moscow) later moved to VECTOR (Novosibirsk, Siberia) and one in the United States (CDC Atlanta). Vaccinations among the general public ceased shortly after the successful eradication campaign, resulting in an increasingly immunologically susceptible population. Because of the possibility of intentional reintroduction of Variola virus and the emergence of other pathogenic poxviruses, there is a great need for the development of medical countermeasures to treat poxvirus disease. It is highly likely that the U.S. FDA “animal rule” will be necessary for regulatory approval of these interventions. Therefore, relevant animal models and the associated supporting assays will require development to stand up to regulatory scrutiny. Methods An optimized real time PCR assay for the detection of orthopoxviruses has been developed by researchers at the United States Army Research Institute of Infectious Diseases (USAMRIID). To support animal studies that will be used to support approval of medical countermeasures by the U.S. FDA, the assay was designed to quantitate poxvirus genomic DNA in a nonhuman primate (cynomolgus macaque) blood matrix as a measurement of viremia. This manuscript describes the validation of the process, including DNA extraction from whole blood anticoagulated with EDTA, for obtaining and quantitating monkeypox genomes by evaluating precision, accuracy, the standard curve, specificity, robustness and stability of the assay and/or components of the assay. Results The assay had a lower limit of quantitation of 50 genome copies/5 uL sample, upper limit of quantitation of 5 × 107 GC/5uL sample and a limit of detection of 2.5 genome copies /5uL sample. The assay was specific for orthopoxvirus. Matrix effects were detected and suggest the presence of PCR inhibitor(s) that was co-extracted with the target DNA. Conclusions The assay has been validated for the purpose of quantitating monkeypox viral load in blood from cynomolgus macaques. This assay has and will continue to support submissions to the FDA for approval of antiviral therapeutics for smallpox. Electronic supplementary material The online version of this article (10.1186/s12985-017-0880-8) contains supplementary material, which is available to authorized users.

Continuous manufacturing (CM) is an emerging technology in the pharmaceutical manufacturing sector, and the understanding of the impact on product quality is currently evolving. As the final purification and isolation step, crystallization has a significant impact on the final physicochemical properties of drug substance and is considered a critical process step in achieving the continuous manufacturing of drug substances. Although many publications previously focused on various innovative techniques to continuously make crystals with desired properties, engineering difficulties such as system design, automation, and integration with process analytical technology (PAT) tools have not been thoroughly discussed. Here, we focus on how to develop a continuous crystallization system, from the perspective of process engineering, and the related risk considerations on product quality. Specifically, we designed and built an automated two-stage mixed suspension mixed product removal (MSMPR) crystallization platfor...

For continuous manufacturing of pharmaceuticals, understanding the dynamics of how a material flows through the process is critical with respect to the development of a control strategy for product quality assurance. Such understanding of the process dynamics can be obtained by characterization of the residence time distribution (RTD). The RTD for a process is not fixed and can vary due to changes in operating conditions or physiochemical properties of the blend. As such the RTD needs to be evaluated over the range of operating condition that can impact process dynamics (e.g. throughput, impeller rotation rate etc.). In this paper, we demonstrate that the dimensionless RTD (normalized with respect to the mean residence time) is invariant with throughput and impeller rotation rates under certain conditions for the two continuous direct compression processes. We present a case study to illustrate the utility of this relationship for predicting the process dynamics at different operating conditions (i.e., throughputs) and evaluating the impact of variations in the process dynamics on the control strategy for a continuous direct compression process.

Given that consumers value quality, and advertising content informs consumers’ beliefs about quality, it is not surprising that high-quality brands emphasize quality in their advertising content. What is less obvious is whether firms with lower-quality brands should also follow suit and emphasize quality in their advertising to signal a higher quality. We examine this issue and study the effectiveness of quality-based advertising messages. Our field study relates brands’ monthly sales to their advertised quality claims across 1,876 print ads in national magazines and Consumer Reports–based product quality ratings over more than two decades. Contrary to the generally held yet erroneous belief in the efficacy of low-quality products emphasizing quality in their advertising, we demonstrate that (1) it is not beneficial for a low-quality firm to emphasize quality in its advertising, and (2) it is effective for a high-quality firm to do so. An analysis of parameter values from a published category-agnostic simulation and an experiment that examines consumers’ responses to quality claims in a second product category yields convergent insights.

The utility of shell overlays to oyster (Crassostrea virginica) plantings as a cownose ray (Rhinoptera bonasus) predator deterrence mechanism was examined. Typical industry practice of oyster seed planting was followed in an experimental design employing treatment areas of 0.5–1.0 acre (0.2–0.4 hectare). Areas were prepared in the Lower Machodoc Creek, Virginia, by the initial application of shell to insure a stable substrate under planted seed oysters. Seed oysters were planted using standard industry methods. Experimental areas were located, two upstream and two downstream, of a constriction in the Lower Machodoc that dictated differing physical environments in the respective locations with downstream locations being more exposed to northeast wind-driven stresses and, historically, a greater incidence of ray predation. Once oysters were planted, two of the areas, one upstream and one downstream of the aforementioned constriction, were additionally treated with a shell overlay as a predation det...

In this paper, we explore how diverse community networks in Brazil have locally advanced seed production and institutional systems to enhance a restoration economy. By focusing on the experiences of the six major native seed suppliers in Amazonia, the Cerrado, and the Atlantic Forest, we estimate the capacity to scale-up community-based systems to meet a large-scale restoration target as a rural development strategy. Over one decade, 1016 collectors traded 416.91 tonnes of native seeds representing, on average, 31.41 kilos yearly and USD 256.5 as household income. Based on this well documented empirical evidence, we estimate that Brazil&rsquo, s restoration goal would require from 3.6 to 15.6 thousand tonnes of native seeds depending on the share of each restoration method adopted with potential work opportunities for 13.2 to 57.1 thousand collectors yearly and total income from USD 34 to 146 million. We argue that community networks represent feasible arrangements for increasing the availability of plant material sources which provide high socio-economic benefits. For scaling up native seed sources, we suggest the following key strategies: (i) government incentives and subsidies, (ii) enforcement of ecosystem restoration, (iii) community participation, (iv) adaptation of the seed regulations, (v) technological development, and (vi) seed market diversification.

While community forestry has shown promise to reduce rural poverty, improve reforestation and potentially offset carbon emissions, many projects have failed, either partly or completely. In order to understand why community forestry succeeds or fails, we examined in detail the literature related to community forestry from three countries, Mexico, Nepal and the Philippines. We also drew on experiences in other countries in Asia, Latin America and Africa. We identified five main interconnected factors which the literature suggests are often critical to the success of community forestry. To integrate the many ways in which community forestry projects can improve the state of these factors, we use the concept of ‘bonding social capital’, i.e. communities’ ability to work together towards a common aim and ‘bridging social capital’, i.e. their ability to liaise with the outside world. To understand the interaction of the five success factors and the way in which improvements to bonding or bridging social capital may affect them, we developed a causal diagram which depicts the interrelationships between the success factors and the key points at which project inputs may be best applied. It is clear from our analysis that failing to appreciate both the complexity and interaction of the various influences may lead to project failure.

The research examines how the attractiveness of children in need affects the empathy they evoke and the subsequent help they receive from unrelated adults. The authors find that attractive children are attributed desirable characteristics related to social competence, which is consistent with the "beautiful is good" stereotype. Ironically, the authors find that these attributions reduce the empathy evoked by attractive children and the help they receive from unrelated adults as long as their need is not severe. These effects are demonstrated in four experiments. The research identifies a significant cost of being beautiful and an important exception to the beautiful is good stereotype. The results also have practical implications for how children are portrayed in promotional materials for disaster relief agencies, children's hospitals, and other charities.

What do American adults believe about what, where, when, how much, and how often it is appropriate to eat? Such normative beliefs originate from family and friends through socialization processes, but they are also influenced by governments, educational institutions, and businesses. Norms therefore provide an important link between the social environment and individual attitudes and behaviors. This paper reports on five studies that identify, develop, and validate measures of normative beliefs about eating. In study 1 we use an inductive method to identify what American adults believe are appropriate or desirable eating behaviors. Studies 2 and 3 are used to purify and assess the discriminant and nomological validity of the proposed set of 18 unidimensional eating norms. Study 4 assesses predictive validity and finds that acting in a norm-consistent fashion is associated with lower Body Mass Index (BMI), and greater body satisfaction and subjective health. Study 5 assesses the underlying social desirability and perceived healthiness of the norms.

Since decades, varieties of amphiphilic polymers have been widely investigated for improving aqueous solubility and bioavailability of the hydrophobic drugs. The upcoming approach is to develop more efficient advanced nano-carrier molecules capable of more than drug delivery. Herein, we report the design and synthesis of some novel carrier molecules with multiple applications including drug encapsulation, drug delivery and diagnosis (imaging). Copolymers were synthesized using dimethyl 5-hydroxy/aminoisophthalate, poly(ethylene glycols) and Candida antarctica lipase (CAL-B, Novozym 435). CAL-B selectively catalyses the trans esterification reaction under solvent less condition using primary hydroxyls of poly(ethylene glycols) and leaving behind phenolic hydroxyl for post polymerization modifications. The obtained copolymers were further tethered with perfluorinated aliphatic chains to make them amphiphilic. The synthesized materials were investigated for their micellar behavior, temperature dependent stability (in aqueous solution), encapsulation capacity, and imaging potential by measuring the sensitivity of these perfluorinated materials towards 19 F NMR in NMR tube. It was observed that perfluorinated amphiphilic copolymers could encapsulate up to 14% (by wt) of hydrophobic drug and showed decent 19 F NMR signals even at a very low concentration. Therefore, these perfluorinated copolymers hold considerable potential for further investigation as advanced nano-carrier molecules for biomedical applications.

We conduct three experiments in which participants in dyads choose between two restaurants, each of which is preferred by only one participant, and one participant has the power to decide which restaurant both will patronize. We find that the power to make a joint decision increases satisfaction with the choice only when those involved have a competitive decision orientation, a weak or anonymous relationship, and the outcome they choose is subsequently available. Participants who have a cooperative orientation are satisfied whether or not they have power and whether or not the resulting choice is consistent with their initial preferences.

To date, proteomic analyses on gastrointestinal cancer tissue samples have been performed using surgical specimens only, which are obtained after a diagnosis is made. To determine if a proteomic signature obtained from endoscopic biopsy samples could be found to assist with diagnosis, frozen endoscopic biopsy samples collected from 63 gastric cancer patients and 43 healthy volunteers were analyzed using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. A statistical classification model was developed to distinguish tumor from normal tissues using half the samples and validated with the other half. A protein profile was discovered consisting of 73 signals that could classify 32 cancer and 22 normal samples in the validation set with high predictive values (positive and negative predictive values for cancer, 96.8% and 91.3%; sensitivity, 93.8%; specificity, 95.5%). Signals overexpressed in tumors were identified as alpha-defensin-1, alpha-defensin-2, calgranulin A, and calgranulin B. A protein profile was also found to distinguish pathologic stage Ia (pT1N0M0) samples (n = 10) from more advanced stage (Ib or higher) tumors (n = 48). Thus, protein profiles obtained from endoscopic biopsy samples may be useful in assisting with the diagnosis of gastric cancer and, possibly, in identifying early stage disease.

The human epidermal growth factor receptors, EGFR and HER2, are members of the EGFR family of cell-surface receptors/tyrosine kinases. EGFR- and HER2-positive cancers represent a more aggressive disease with greater likelihood of recurrence, poorer prognosis, and decreased survival rate, compared to EGFR- or HER2-negative cancers. The details of HER2 proto-oncogenic functions are not deeply understood, partially because of a restricted availability of tools for EGFR and HER2 detection (A. Sorkin and L. K. Goh, Exp. Cell Res. 2009, 315, 683-696). We have created photostable and relatively simple-to-produce imaging probes for in vitro staining of EGFR and HER2. These new reagents, called affiprobes, consist of a targeting moiety, a HER2- or EGFR-specific Affibody molecule, and a fluorescent moiety, mCherry (red) or EGFP (green). Our flow cytometry and confocal microscopy experiments demonstrated high specificity and signal/background ratio of affiprobes. Affiprobes are able to stain both live cells and frozen tumor xenograph sections. This type of optical probe can easily be extended for targeting other cell-surface antigens/ receptors.

Viral release is a necessary component of HIV-1 replication. To be an efficient process, viral budding relies on recruitment of the ubiquitin E2 variant (UEV) domain of the human tumor susceptibility gene 101 protein (Tsg101) by major structural proteins of HIV-1.1,2 This entails the direct interaction of the Tsg101 UEV domain with a proline rich motif (PRM) within the viral Gagp6 protein that contains a conserved sequence, Pro-Thr/Ser-Ala-Pro [“P(T/S)AP”].3,4 Over-expression of the Tsg101 UEV inhibits virus release by interfering with processing of the p6 domains and this effect is abrogated by mutation within the P(T/S)AP binding site. In theory, blocking this critical Tsg101-p6 interaction could prevent viral budding and provide a basis for new targeted antiretroviral therapies.5,6 This is supported by the recent finding that inhibition of HIV budding can be achieved by cyclic peptides that interfere with Tsg101-Gag interactions.7 In an effort to develop Tsg101-binding inhibitors using the reported p6-derived 9-mer wild-type (WT) sequence “P1E2P3T4A5P6P7E8E9,” we had previously improved Tsg101-binding affinity by applying hydrazone- and hydrazide-library techniques.8 We also reported an oxime-based post solid-phase diversification approach to peptide library construction.9–11 However, the peptide mimetics resulting from this earlier work showed poor biovailability in cell-based experiments. Because poor cellular uptake was thought to contribute to the low bioavailability, peptides were conjugated with known membrane carrier antennapedia and HIV-Tat (48–60) peptides.12 Although these constructs did exhibit apparent enhanced cellular uptake, the Tsg101 binding affinity of the conjugates was seriously disrupted. Therefore, we sought an alternate approach that could improve cell bioavailability while maintaining Tsg101-binding affinity. We took note of the fact that cyclic versions of linear peptides can result in better cell-permeability. Additionally, appropriately restricting solution conformations through cyclization can also afford higher affinity.13–17 The objective of the current study was to apply RCM strategies to the WT 9-mer peptide and to observe the effects that such macrocyclization would have on Tsg101-binding affinity and cellular bioavailability.

Blocking the interaction between phosphotyrosine (pTyr)-containing activated receptors and the Src homology 2 (SH2) domain of the growth factor receptor-bound protein 2 (Grb 2) is considered to be an effective and non-cytotoxic strategy to develop new anti-proliferate agents due to its potential to shut down the Ras activation pathway. In this study, a series of phosphotyrosine containing cyclic pentapeptides were designed and synthesized based upon the phage library derived cyclopeptide, G1TE. A comprehensive SAR study was also carried out to develop potent Grb2-SH2 domain antagonists based upon this novel template. With both the peptidomimetic optimization of the amino acid side-chains and the constraint of the backbone conformation guided by molecular modeling, we developed several potent antagonists with low micromolar range binding affinity, such as cyclic peptide 15 with an Kd = 0.359 μM, which is providing a novel template for the development of Grb2-SH2 domain antagonists as potential therapeutics for certain cancers.

Expression of the retroviral Gag protein leads to formation of virus-like particles in mammalian cells. In vitro and in vivo experiments show that nucleic acid is also required for particle assembly. However, several studies have demonstrated that chimeric proteins in which the nucleocapsid domain of Gag is replaced by a leucine zipper motif can also assemble efficiently in mammalian cells. We have now analyzed assembly by chimeric proteins in which nucleocapsid of human immunodeficiency virus type 1 (HIV-1) Gag is replaced by either a dimerizing or a trimerizing zipper. Both proteins assemble well in human 293T cells; the released particles lack detectable RNA. The proteins can coassemble into particles together with full-length, wild-type Gag. We purified these proteins from bacterial lysates. These recombinant “Gag-Zipper” proteins are oligomeric in solution and do not assemble unless cofactors are added; either nucleic acid or inositol phosphates (IPs) can promote particle assembly. When mixed with one equivalent of IPs (which do not support assembly of wild-type Gag), the “dimerizing” Gag-Zipper protein misassembles into very small particles, while the “trimerizing” protein assembles correctly. However, addition of both IPs and nucleic acid leads to correct assembly of all three proteins; the “dimerizing” Gag-Zipper protein also assembles correctly if inositol hexakisphosphate is supplemented with other polyanions. We suggest that correct assembly requires both oligomeric association at the C terminus of Gag and neutralization of positive charges near its N terminus.

Hydrophobic active pharmaceutical ingredients (APIs) are often difficult to deliver effectively because of formulation limitations. Nanosuspensions of such drugs may be used to increase bioavailability and offer a variety of delivery options including injection, inhalation, oral, and transdermal. Microfluidics reaction technology (MRT) was used successfully to produce submicrometer API suspensions via a continuous process that involves solvent/antisolvent crystallization. As proof of concept, nanosuspensions of norfloxacin (NFN), an antibacterial agent, were produced varying the key parameters of the technology. The nanosuspensions had narrow particle size distributions and median particle sizes in the range of 170-350 nm. The particle size depends on the supersaturation ratio and energy dissipation expressed as processing pressure. However, the particle size was found to be insensitive to the presence of the surfactant used. The crystalline structure of NFN was not affected by the mixing intensity but by the solvent/antisolvent system. This "bottom up" process for creating nanosuspensions was compared to a "top down" process, in which NFN nanosuspensions were created as a result of particle size reduction. It was found that the "bottom up" process was substantially more efficient and resulted in smaller particles than the "top down" process. MRT is based on an impinging jet reactor design with jet velocities and energy dissipation that is orders of magnitude higher than those of conventional impinging jet reactors. The technology provides precise control of the feed rates and the subsequent location and intensity of mixing of the reactants. It may be the best choice economically due to its process intensification character that minimizes energy requirements and the proven scalability of the reactor.

The research examines viewers' actual responses to four televised fund-raising drives by a public television station over a 2-year period. The 584 pledge breaks we studied contain 4,868 individual appeals that were decomposed into two underlying dimensions based on the empathy-helping hypothesis: the appeal beneficiary (self versus other) and emotional valence (positive versus negative). We find that the most effective fund-raising appeals communicate the benefits to others rather than to the self and evoke negative rather than positive emotions. Appeals that emphasize benefits to the self significantly reduce the number of calls to the station, particularly when they have a positive emotional valence.

The approval of major infrastructure and industrial developments is often dependent on the results of environmental and social impact assessments (EIA and SIA, respectively). Depending on the recommendations of ‘experts’ undertaking the assessments, projects are approved, rejected or modified to take account of unintended negative consequences. In practice, there are pressures on consultants to come up with favourable results, and unfavourable results are often ignored. These pressures may be subtle. For example, because consultants are usually employed by somebody with an interest in seeing projects go ahead, any consultant with a reputation for being too negative is unlikely to get work. The present paper explores issues faced by anthropologists in SIA, as well as the structure of interests within which SIA is carried out, and ways in which ethical dilemmas can be negotiated. It asks whether simply opting out of the process altogether is an ethical option preferable to critical engagement.

HIV-1 viral assembly requires a direct interaction between a Pro-Thr-Ala-Pro (“PTAP”) motif in the viral protein Gag-p6 and the cellular endosomal sorting factor Tsg101. In an effort to develop competitive inhibitors of this interaction, an SAR study was conducted based on the application of post solid-phase oxime formation involving the sequential insertion of aminooxy-containing residues within a nonamer parent peptide followed by reaction with libraries of aldehydes. Approximately 15–20-fold enhancement in binding affinity was achieved by this approach.

In this paper we address the often sterile and circular debates over relationships between poverty and deforestation. These debates revolve around questions of whether forest loss causes poverty or poverty contributes to forest encroachment, and questions of whether it is loss of access to forests or dependence on forest-based livelihoods that cause poverty. We suggest that a way beyond the impasse is to set such debates within the context of agrarian change. Livelihoods of those who live in or near forests depend considerably on a rapidly changing agriculture, yet agrarian contexts receive only background attention in popular, political and academic discourse over poverty and forests. Moreover, to the extent that agriculture is considered, little heed is paid to social, technical and economic change. We therefore address agriculture's changing relationships with the wider economy, otherwise referred to as the agrarian transition, and with the natural resource base on which it depends. The paper draws on the experience of Thailand to illustrate our key argument, and more specifically addresses the situation on the resource periphery through a look at the agriculture-forest interface.

The stereoselective synthesis of aminooxy-containing proline analogues bearing Fmoc/Boc or Fmoc/Mtt protection that renders them suitable for incorporation into peptides using Fmoc protocols is reported. Acid-catalyzed unmasking at the completion of peptide synthesis yields free aminooxy-functionalities for oxime formation through reaction with libraries of aldehydes. This allows post solid-phase diversification strategies that may facilitate structure–activity relationship studies.

Ring-closing metathesis (RCM) was employed to join carboxy-terminal alkenyl glycine side chains together with vinyl- and allyl-functionality appended to the beta-methylene of amino-terminal phosphotyrosyl (pTyr) mimetics. This required the synthesis of a variety of new pTyr mimetics, including a novel aza-containing analogue. Many of the resulting 15-member macrocyclic tetrapeptide mimetics exhibited low nanomolar Grb2 SH2 domain-binding affinities in spite of the fact that differing ring junction stereochemistries and geometries of the RCM-derived double bond were employed. The finding that significant latitude exists in the structural requirements for ring closure may facilitate the development of therapeutically relevant macrocyle-based Grb2 SH2 domain-binding antagonists. The synthetic approaches used in this study may also find application to peptide mimetics directed at other biological targets.

Individual protein binding sites on DNA can be measured in bits of information. This information is related to the free energy of binding by the second law of thermodynamics, but binding kinetics appear to be inaccessible from sequence information since the relative contributions of the on- and off-rates to the binding constant, and hence the free energy, are unknown. However, the on-rate could be independent of the sequence since a protein is likely to bind once it is near a site. To test this, we used surface plasmon resonance and electromobility shift assays to determine the kinetics for binding of the Fis protein to a range of naturally occurring binding sites. We observed that the logarithm of the off-rate is indeed proportional to the individual information of the binding sites, as predicted. However, the on-rate is also related to the information, but to a lesser degree. We suggest that the on-rate is mostly determined by DNA bending, which in turn is determined by the sequence information. Finally, we observed a break in the binding curve around zero bits of information. The break is expected from information theory because it represents the coding demarcation between specific and nonspecific binding.

By enzymatically hydrolyzing the terminal phosphodiester bond at the 3'-ends of DNA breaks, tyrosyl-DNA phosphodiesterase (Tdp1) repairs topoisomerase-DNA covalent complexes and processes the DNA ends for DNA repair. To identify novel Tdp1 inhibitors, we developed a high-throughput assay that uses electrochemiluminescent (ECL) substrates. Subsequent to screening of 1981 compounds from the 'diversity set' of the NCI-Developmental Therapeutics Program, here we report that furamidine inhibits Tdp1 at low micromolar concentrations. Inhibition of Tdp1 by furamidine is effective both with single- and double-stranded substrates but is slightly stronger with the duplex DNA. Surface plasmon resonance studies show that furamidine binds both single- and double-stranded DNA, though more weakly with the single-stranded substrate DNA. Thus, the inhibition of Tdp1 activity could in part be due to the binding of furamidine to DNA. However, the inhibition of Tdp1 by furamidine is independent of the substrate DNA sequence. The kinetics of Tdp1 inhibition by furamidine was influenced by the drug to enzyme ratio and duration of the reaction. Comparison with related dications shows that furamidine inhibits Tdp1 more effectively than berenil, while pentamidine was inactive. Thus, furamidine represents the most potent Tdp1 inhibitor reported to date.

Copper (I) promoted [3+2] Huisgen cycloaddition of azides with terminal alkynes was used to prepare triazole-containing macrocycles based on the Grb2 SH2 domain-binding motif, ‘Pmp-Ac6c-Asn’, where Pmp and Ac6c stand for 4-phosphonomethylphenylalanine and 1-aminocyclohexanecarboxylic acid, respectively. When cycloaddition reactions were conducted at 1 mM substrate concentrations, cyclization of monomeric units occurred. At 2 mM substrate concentrations the predominant products were macrocyclic dimers. In Grb2 SH2 domain-binding assays the monomeric (S)-Pmp-containing macrocycle exhibited a Kd value of 0.23 μM, while the corresponding dimeric macrocycle was found to have greater than 50-fold higher affinity. The open-chain dimer was also found to have affinity equal to the dimeric macrocycle. This work represents the first application of ‘click chemistry’ to the synthesis of SH2 domain-binding inhibitors and indicates its potential utility.

Mass transfer experiments were conducted to determine the transport and interaction parameters of selected molecules in hydrated bacterial cellulose (BC) membranes. The objective was to determine physiochemical characteristics and elucidate the mechanisms governing transport in relation to the membrane structure. Pore and sorption models developed previously for the analysis of transport in hydrogel membranes were relevant to the cellulose membrane system, including: (1) interfacial phenomena between the bulk fluid and the outer membrane surfaces and/or along a pore wall, (2) sorption into the membrane matrix itself with diffusion possibly affected by immobilization at specific interactive sites, (3) free and/or fixed site diffusion within the matrix and if appropriate, through the porous regions, whether as distinct pores, micro-channels or other non-homogeneous/discrete areas and (4) chemical reactions that could alter the nature of the diffusing species or the media itself. Since all these mechanisms may be active, our classification, and thus characterization, was based upon whichever mechanism dominates. The diffusion coefficients of various dextrans (44–260 kDa), measured using a horizontal flow diffusion chamber, were compared to their corresponding values in water to demonstrate hindered diffusion. The degree of hindrance, ψ, was the same for all four dextrans indicating that the parameter is not a function of pore or micro-channel size but rather due to the presence of membrane fibers. Three representative marker molecules (Vitamin B12, lysozyme and bovine serum albumin) were evaluated using the same apparatus combined with desorption experiments to measure permeation (P) and effective diffusion (Deff) coefficients. Partition coefficients, H, were subsequently calculated and verified experimentally. Estimates of Deff in the BC membranes were made through a weighted average using literature values for the diffusion of the three molecules in water and regenerated cellulose along with measured H and ψ values. The experimental results are in excellent agreement with these estimates indicating the presence of dual transport mechanisms, for solute transport through the continuous water phase and cellulose matrix, with some hindrance of molecular diffusion via fiber obstruction. With Vitamin B12 and lysozyme (the two smallest solutes), equilibrium interactions such as adsorption and solubility are also important. These results help clarify the potential utility of these novel bio-derived membranes in a variety of possible separations scenarios.

Crosslinked hyaluronan (HA) hydrogels preloaded with two cytokine growth factors, vascular endothelial growth factor (VEGF) and keratinocyte growth factor (KGF), were employed to elicit new microvessel growth in vivo. As a major glycosaminoglycan (GAG) component of extracellular matrix (ECM), HA is an excellent biopolymeric building block for new biomimetic, biocompatible therapeutic materials. HA hydrogel film samples were surgically implanted in the ear pinnae of mice, and the ears were harvested at 7 or 14 days post-implantation. Histologic analysis showed that each of the groups receiving an implant demonstrated significantly more microvessel density than control ears undergoing surgery but receiving no implant (p

Gray market activity has become increasingly prevalent. The prevailing wisdom in marketing is to use more severe enforcement to deter gray marketing. However, the certainty and speed of enforcement may also have a bearing on the incidence of violations. This article examines whether and how enforcement deters gray marketing. The results from a field survey of manufacturers and an experimental design suggest that, by itself, enforcement severity has no impact. Deterrence results only when the multiple facets of enforcement are used in combination.

This research examines the effects of relationship quality (RQ) on customers' desires to retaliate after service failures. We posit that the effects of RQ are contingent upon the attributions customers make about the firm's controllability over a service failure. Two competing hypotheses are examined and reconciled. The "love is blind" hypothesis posits that when low controllability is inferred, high RQ customers experience a lesser desire for retaliation than low RQ customers. On the other hand, the "love becomes hate" hypothesis specifies that when high controllability is inferred, high RQ customers experience a greater desire for retaliation than low RQ customers. The hypotheses are tested with a survey- based design and a partial least squares (PLS) model that incorporates a multiplicative latent construct.

The research develops and tests a model of gender differences on the effects of competition and cooperation on decision satisfaction. It was hypothesized and found that males’ satisfaction is dependent on their ability to impose their preferences on their female counterpart within a mixed-gender dyad. In contrast, females’ satisfaction is only affected by the degree to which dyad members behave cooperatively. The model is tested within the context of 76 mixed-gender dyads that are in an established relationship. The dyads are sampled from shoppers at a national retailer, and report on their decision-making process as they exit the store. A partial-least-squares (PLS) methodology is used to test for differences in the effects of cooperative and competitive behaviors on males’ and females’ satisfaction with a dyadic decision. © 2006 Wiley Periodicals, Inc.

Although considerable effort has been devoted to developing Grb2 SH2 domain-binding antagonists, important questions related to ligand specificity, and identification of intracellular targets remain unanswered. In order to begin addressing these issues, the design, synthesis, and evaluation of a novel biotinylated macrocycle are reported that bears biotin functionality at a C-terminal rather than the traditional N-terminal position. With a Grb2 SH2 domain-binding K eq value of 3.4 nM, the title macrocycle ( 5 ) is among the most potent biotinylated SH2 domain-binding ligands yet disclosed. This should be a useful tool for elucidating physiological targets of certain Grb2 SH2 domain-binding antagonists.

MacKenzie, Lutz, and Belch (1986) test four advertising attitude models and find that the Dual Mediation Hypothesis is the best. This research proposes an extended model within an online context, using intentions to return (Ir) to a Web site versus purchase intentions, with a direct path between attitudes toward the Web site (Asite) and Ir. This path is hypothesized as Web sites contain informative or entertaining content that attracts subsequent visits, and Ir depends on other non-brand-related factors such as security, ease of use, transactional capabilities, etc. Data from visitors to three actual Web sites––digital cameras, watches, and a charity––demonstrate significant relationships between Asite and Ir. In support of this perspective, when Asite was decomposed into its claim and non-claim components, the non-claim component had a significant effect on Ir for all three sites. Implications for online researchers and advertisers are discussed.

As typified by 2-[(9S,10S,14R,18S)-18-(2-amino-2-oxoethyl)-14-[(5-methyl-1H-indol-l-yl)methyl]-8,17,20-trioxo-10-[4-(phosphonomethyl)phenyl]-7,16,19-triazaspiro[5.14]icos-11-en-9-yl)acetic acid ((14R)-1b), ring-closing methathesis-derived macrocyclic tetrapeptide mimetics have recently been reported that bind with high affinity to Grb2 SH2 domains in both extracellular and whole-cell assays. The synthetic complexity of this class of agents limits further therapeutic development. Although a significant component of this synthetic complexity arises from the presence of three stereogenic centers, C(9) (S), C(10) (S), and C(14) (R), it is unclear whether stereoselective introduction of defined configuration at C(14) is required for high-affinity binding. Reported herein is a synthetic route to these macrocycles lacking stereoselectivity in the formation of the C(14) ring junction, which is four synthetic steps shorter than the original stereoselective synthesis. Separation of C(14)-epimers obtained by this approach was achieved by preparative HPLC. Molecular-dynamics studies of ligands bound to the Grb2 SH2 domain protein indicated that the (14R)-configuration should display more-favorable interactions with the protein relative to the (14S)-epimer. Indeed, although surface-plasmon-resonance-derived binding constants to Grb2 SH2 domain protein indicated that the affinity of the (14R)-epimer (K D = 4.8 nM) is greater than that of the (14S)-epimer (K D = 11 nM), it is only marginally so. Therefore, little affinity would be lost through a non-stereoselective synthesis of the C(14)-center. Further studies are in progress to explore reduced structural complexity at the C(14)-center.

Two studies examine gender differences in responses to advertising with emotional content that varies on agency—a fundamental component of the male, but not female, stereotype. As hypothesized, males reported a less pleasant viewing experience and a less favorable attitude toward the advertisement (Aad) when a low‐agency emotion (i.e., stereotype‐incongruent) ad was viewed with another male, while their responses were not affected by the presence of another person when they were exposed to a high‐agency emotion (i.e., stereotype‐congruent) ad. Males’ and females’ private responses were not significantly different, and females’ responses were invariant across social contexts and type of ad.

The relationship between attitude toward the advertisement (A ad ) and intentions to buy (I b ) is a controversial one. The present research examines the potential for a direct relationship between A ad and I b within an on-line advertising context, substituting A site (attitude toward the Web site) for A ad . The article replicates previous findings with respect to the four competing A ad models they tested. The article then predicts and finds a significant A site → I b path based on an opportunity and motivation perspective. In terms of opportunity, Web sites contain nonproduct information that is independent of traditional measures of brand attitude (A b ), such as security, ease of use, transactional capabilities, which are likely to affect I b . Further, on-line consumers are motivated to process this information when they visit a Web site because they cannot directly examine the products they are considering. Evidence for the opportunity and motivation perspective is provided by the decomposition of A site into its claim and nonclaim components and assessment of the A site → I b path across levels of motivation to process. Implications for on-line advertisers are discussed. © 2005 Wiley Periodicals, Inc.

To gauge the experimental variability associated with Biacore analysis, 36 different investigators analyzed a small molecule/enzyme interaction under similar conditions. Acetazolamide (222 g/mol) binding to carbonic anhydrase II (CAII; 30,000 Da) was chosen as a model system. Both reagents were stable and their interaction posed a challenge to measure because of the low molecular weight of the analyte and the fast association rate constant. Each investigator created three different density surfaces of CAII and analyzed an identical dilution series of acetazolamide (ranging from 4.1 to 1000 nM). The greatest variability in the results was observed during the enzyme immobilization step since each investigator provided their own surface activating reagents. Variability in the quality of the acetazolamide binding responses was likely a product of how well the investigators’ instruments had been maintained. To determine the reaction kinetics, the responses from the different density surfaces were fit globally to a 1:1 interaction model that included a term for mass transport. The averaged association and dissociation rate constants were 3.1 ± 1.6 × 10 6 M −1 s −1 and 6.7 ± 2.5 × 10 −2 s −1 , respectively, which corresponded to an average equilibrium dissociation constant ( K D ) of 2.6 ± 1.4 × 10 −8 M. The results provide a benchmark of variability in interpreting binding constants from the biosensor and highlight keys areas that should be considered when analyzing small molecule interactions.

The human immunodeficiency virus (HIV) epidemic is an important medical problem. Although combination drug regimens have produced dramatic decreases in viral load, current therapies do not provide a cure for HIV infection. We have used structure-based design and combinatorial medicinal chemistry to identify potent and selective HIV-1 reverse transcriptase (RT) inhibitors that may work by a mechanism distinct from that of current HIV drugs. The most potent of these compounds (compound 4 , 2-naphthalenesulfonic acid, 4-hydroxy-7-[[[[5-hydroxy-6-[(4-cinnamylphenyl)azo]-7-sulfo-2-naphthalenyl]amino]carbonyl]amino]-3-[(4-cinnamylphenyl)azo], disodium salt) has an IC 50 of 90 nM for inhibition of polymerase chain extension, a K d of 40 nM for inhibition of DNA–RT binding, and an IC 50 of 25–100 nM for inhibition of RNaseH cleavage. The parent compound ( 1 ) was as effective against 10 nucleoside and non-nucleoside resistant HIV-1 RT mutants as it was against the wild-type enzyme. Compound 4 inhibited HIV-1 RT and murine leukemia virus (MLV) RT, but it did not inhibit T 4 DNA polymerase, T 7 DNA polymerase, or the Klenow fragment at concentrations up to 200 nM. Finally, compound 4 protected cells from HIV-1 infection at a concentration more than 40 times lower than the concentration at which it caused cellular toxicity.

Unlike prior research that treats social-desirability bias (SDB) as measure contamination, the present research asserts that significant associations between measures of SDB and value self-reports are evidence of measure validity. The degree to which value self-reports are influenced by SDB also reflects the relative importance of values within a culture. Values that are most important have the greatest self-presentational implications and therefore should be most affected by SDB. Moreover, differences between raw and SDB-corrected value self-reports indicate the extent to which values are personal (i.e., private) or public in nature. The research is based on two national samples of American adults 18 years of age and older. Implications for research on values are discussed. © 2000 John Wiley & Sons, Inc.

Host proteins are incorporated both on and inside human immunodeficiency virus type 1 (HIV-1) virions. To identify cellular proteins inside HIV-1, virion preparations were treated by a protease-digestion technique that removes external host proteins, allowing for the study of the proteins inside the virus. Treated HIV-1 preparations were analyzed by immunoblot, high-pressure liquid chromatography, and protein sequence analyses. These analyses identified several cellular proteins inside HIV-1: elongation factor 1α, glyceraldehyde-3-phosphate dehydrogenase, HS-1, phosphatidylethanolamine-binding protein, Pin1, Lck, Nm23-H1, and the C-terminal tail of CD43. Several of these proteins were found as fragments of their full-sized proteins that appear to be generated by our protease treatment of the virions, the HIV-1 protease, or a cellular protease. Recent advances in cell biology and biochemistry have identified some of these proteins as actin-binding proteins. These results support the hypothesis that actin filaments are incorporated into the virion and may provide additional clues for the understanding of the interaction between viral and cellular proteins during assembly and budding.

The significance of volunteering for both individuals and society has lead to numerous studies on this behavior across the social sciences. However, virtually no prior research has evaluated how and to what extent organizations can effectively encourage individuals to contribute time to a worthy cause. The present research uses a social norm perspective to examine the conditions under which promotional appeals based on group need and promises of recognition affect volunteerism. The perspective suggests that norm compliance can be expected only when the prescribed behavior is both important to the group's welfare and subject to group-mediated rewards. Consequently, we hypothesize that promotional appeals based on group need and promised recognition are effective only when they are used in combination. Results of a laboratory and a field experiment are consistent with this hypothesis and provide insights into the process by which the appeals affect individuals' decisions to help. The results also have implications for understanding and promoting other socially desirable behaviors such as recycling, energy conservation, litter reduction, and the purchase of "green" products. Copyright 1998 by the University of Chicago.

Why do fans of losing sports teams, alumni of poorly ranked educational programs, and patrons of charities that are rocked by scandal maintain or even increase their support? The present research investigates this issue by examining how differences in group success affect the factors that lead to identification and ultimately the incidence of group-supportive behaviors. The results of a two-group field study of professional sports fans suggest that members of unsuccessful groups identify on the basis of their involvement with the domain in which the group operates and the attractiveness of group members. Also, whereas perceived group performance is the most important factor leading to identification for members of successful groups, this factor is irrelevant to members of unsuccessful groups. In addition, the study finds a strong association between the strength of identification and the incidence of group-supportive consumption behaviors for members of both types of groups. The research has important implications for organizations that depend upon member support, such as sports teams, educational institutions, musical groups, and charities. © 1998 John Wiley & Sons, Inc.

Capillary electrophoresis with a polyacrylamide-coated capillary was used for the separation of proteins, and the study of protein-small molecule and protein-DNA interactions. The coating (10% T polyacrylamide) is of sufficient thickness and hydrophilicity to reduce protein adsorption and eliminate electroosmotic flow. Data on the effect of sample solvent constituents on the peak shape and position of protein solutes was investigated. The results show that CE is a relevant and fast technique for the study of protein-DNA and protein-drug interaction. Also, the addition of TRIS-buffer to the sample solution resulted in the CE focusing of some basic proteins, while others were not affected.

One limitation of prior research in channel power is the absence of a conceptual framework which provides insights into the potential for firms to influence intra-channel power relations. This article develops the concept of customer-based channel power and introduces attribution theory as a framework to provide insights into channel power relationships. Specifically, channel members' marketing strategies are shown to influence consumer attributions which, in turn, influence the customer-based channel power of the respective channel members. The issue is important because firms with higher levels of channel power increase their ability to influence marketing decisions of other channel members and may more effectively implement their own strategies. Propositions are developed as guidelines for future research.

Indirect (i.e., structured projective) questioning has been employed frequently in marketing and other social sciences to reduce social desirability bias, that is, systematic error in self-report measures resulting from the desire of respondents to avoid embarrassment and project a favorable image to others. Yet little is known about the validity of indirect questioning in reducing social desirability bias. This article reports on three studies that examine indirect questioning as a technique to reduce social desirability bias on self-report measures. The effects of asking indirect (i.e., structured, projective) questions were compared with direct (i.e., structured, personal) questions. The pattern of results indicates that indirect questioning reduces social desirability bias on variables subject to social influence and has no significant effect on socially neutral variables. The social nature of the differences between direct and indirect questioning groups, and the attribution of an undesirable trait to an out-group but not an in-group target, supports the view that subjects projected their beliefs and evaluations in the indirect response situation. These results are consistent across several product categories and indirect question wordings.

The social context of new product adoption behavior is a key aspect of the diffusion of innovations. Yet little is known about the process by which social contextual factors influence individual adoption decisions. This research develops and tests a model of the effects of perceived consumption visibility and superorder group influence on new product intention formation. A structural equation model is used in an experimental design to provide a comprehensive view of variable interval and to incorporate measurement error. Key findings indicate that (1) perceived visibility of consumption significantly affects consumers' predictions of normative outcomes (i.e., social approval from referents) and (2) superorder group influence has a direct effect on consumers' perceptions of consumption visibility and expectations of both personal (i.e., intrinsically valued product benefits) and normative outcomes from early adoption. The results have important implications for understanding the role of consumption symbols as mechanisms for social differentiation and integration. Copyright 1992 by the University of Chicago.