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Whether formal regional lymph node (LN) evaluation is necessary for patients with appendiceal adenocarcinoma (AA) who have peritoneal metastases is unclear. The aim of this study was to evaluate the prognostic value of LN metastases on survival in patients treated with cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (CRS-HIPEC).A retrospective analysis of the US HIPEC collaborative, a multi-institutional consortium comprising 12 high-volume centers, was performed to identify patients with AA who underwent CRS-HIPEC with adequate LN sampling (≥ 12 LNs).Two hundred-fifty patients with AA who underwent CRS-HIPEC were included. Outcomes were compared between LN - and LN + disease. Baseline patient characteristics between groups were similar, with most patients undergoing complete cytoreduction (0/1: 86.0% vs. 76.8%, p = 0.08), respectively. More adverse tumor factors were found in patients with LN + disease, including poor differentiation, signet ring cells, and lymphovascular invasion. Multivariate analysis of overall survival (OS) found LN + disease was independently associated with worse OS (HR: 2.82 95%CI: 1.25-6.34, p = 0.01), even after correction for receipt of systemic therapy. On Kaplan-Meier analysis, median OS was lower in patients with LN + disease (25.9 months vs. 91.4 months, p 0.01). LN + disease remained associated with poor OS following propensity score matching (HR: 4.98 95%CI: 1.72-14.40, p 0.01) and in patients with PCI ≥ 20 (HR: 3.68 95%CI: 1.54-8.80, p 0.01).In this large multi-institutional study of patients with AA undergoing CRS-HIPEC, LN status remained associated with worse OS even in the setting of advanced peritoneal carcinomatosis. Formal LN evaluation should be performed for most patients with AA undergoing CRS-HIPEC.

The long-term prognosis of patients who undergo cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) for peritoneal surface malignancies (PSM) varies considerably on the basis of histological and operative factors. While overall survival (OS) estimates are used to inform adjuvant therapy and surveillance strategies, conditional survival may provide more clinically relevant estimates of prognosis by accounting for disease-free time elapsed.All patients from 12 academic institutions who underwent CRS ± HIPEC for PSM from 2000 to 2017 were retrospectively analyzed. OS and disease-free survival (DFS) rates were calculated using the Kaplan-Meier method while conditional overall (COS) and conditional disease-free survival (CDFS) rates were calculated at 1, 2, or 3 years from surgery for different tumor histologies.Overall, 1610 patients underwent CRS ± HIPEC. Among patients with benign appendiceal mucinous tumors (N = 460), 5-year OS and COS at 3 years were 92.1% and 96.3% (Δ4.2%), respectively. For patients with well-differentiated appendiceal cancers (N = 400), 5-year OS and COS at 3 years were 76.3% and 88.3% (Δ12.0%), respectively. For patients with high-grade appendiceal cancers (N = 258), 5-year OS and COS at 3 years were 43.8% and 75.4% (Δ31.6%), respectively. For patients with colorectal cancers (N = 362), 5-year OS and COS at 3 years were 31.8% and 67.3% (Δ35.5%), respectively. For patients with peritoneal mesothelioma (N = 130), 5-year OS and COS at 3 years were 67.6% and 89.7% (Δ22.1%), respectively. Similar trends were observed for DFS/CDFS.The conditional survival of patients undergoing CRS ± HIPEC for PSM is associated with tumor histology. COS and CDFS provide a more accurate, dynamic estimate of survival than OS and DFS, especially for patients with more aggressive histologies.

Parkinson’s disease (PD) is the second most common neurodegenerative disorder which requires a long-term, interdisciplinary disease management. While there remains no cure for Parkinson’s disease, treatments are available to help reduce the main symptoms and maintain quality of life for as long as possible. Owing to the global burden faced by chronic conditions such as PD, Assistive technologies (AT’s) are becoming an increasingly common prescribed form of treatment. Low adoption is hampering the potential of digital technologies within health and social care. It is then necessary to employ classification algorithms have been developed for differentiating adopters and non-adopters of these technologies; thereby, potential negative effects on people with PD and cost overruns can be further minimized. This paper bridges this gap by extending the Multi-criteria decision-making approach adopted in technology adoption modeling for people with dementia. First, the fuzzy Analytic Hierarchy Process (FAHP) is applied to estimate the initial relative weights of criteria and sub-criteria. Then, the Decision-making Trial and Evaluation Laboratory (DEMATEL) is used for evaluating the interrelations and feedback among criteria and sub-criteria. The Technique for Order of Preferences by Similarity to Ideal Solution (TOPSIS) is finally implemented to rank three classifiers (Lazy IBk – knearest neighbors, Naive bayes, and J48 decision tree) according to their ability to model technology adoption. A real case study considering is presented to validate the proposed approach.

The use of nanometer-sized semiconductor crystals, known as quantum dots, allows us to directly observe individual biomolecular transactions through a fluorescence microscope. Here, we review the evolution of single quantum dot tracking over the past two decades, highlight key biophysical discoveries facilitated by quantum dots, briefly discuss biochemical and optical implementation strategies for a single quantum dot tracking experiment, and report recent accomplishments of our group at the interface of molecular neuroscience and nanoscience.

A subset of pancreatic body and tail cancers present with locally advanced disease due to involvement of the celiac axis. Previously considered unresectable, these T4 tumors may be extirpated with a distal pancreatectomy and en bloc resection of the celiac trunk in carefully selected patients. In the setting of multimodality treatment, these resections can yield survival similar to resectable and borderline resectable lesions. Robotic surgery has been shown to be safe and feasible in complex pancreatic resections. This article summarizes our patient selection criteria and operative approach to robotic distal pancreatectomy with celiac axis resection (DP-CAR) for locally advanced body and tail tumors of the pancreas.

Mary Ann Beavis, Jesus and Utopia: Looking for the Kingdom of God in the Roman World, reviewed by Alicia Batten Eve-Marie Becker and Peter Pilhofer, eds., Biographie und Persönlichkeit des Paulus, reviewed by Murray Baker Warren Carter, John: Storyteller, Interpreter, Evangelist, reviewed by David J. Hawkin David Catchpole, Jesus People: The Historical Jesus and the Beginnings of Community, reviewed by Agnes Choi Terence E. Fretheim, God and World in the Old Testament: A Relational Theology of Creation, reviewed by William H. Irwin Simon J. Gathercole, The Pre-existent Son: Recovering the Christogies of Matthew, Mark, and Luke, reviewed by Terence L. Donaldson Michael J. Gorman, Apostle of the Crucified Lord. A Theological Introduction to Paul & His Letters, reviewed by Scott M. Lewis Robert C. Hill, Reading the Old Testament in Antioch, reviewed by Shawn W.J. Keough Leander E. Keck, Romans, reviewed by L. Ann Jervis Paul Lawrence, Alan Millard, Heinrich von Siebenthal and John Walton, eds., The IVP Atlas of Bible History, reviewed by 3 Anson F. Rainey and R. Steven Notley, The Sacred Bridge: Carta s Atlas of the Biblical World, reviewed by Brian P. Irwin Peter Leithart, l & 2 Kings, reviewed by Lissa M. Wray Beal Moises Mayordomo, Argumentiert Paulus logisch? Eine Analyse vor dem Hintergrund antiker Logik, reviewed by Ian H. Henderson Stanley E. Porter, ed., Hearing the Old Testament in ,the New Testament, reviewed by Ian W. Scott Willard M. Swartley, Covenant of Peace: The Missing Peace in New Testament Theology and Ethics, reviewed by Gordon Zerbe John H. Walton, Ancient Near Eastern Thought and the Old Testament: Introducing the Conceptual World of the Hebrew Bible, reviewed by Jonathan S. Greer Stephen Westerholm, Understanding Matthew: The Early Christian Worldview of the First Gospel, reviewed by Adrian M. Leske Ritva H. Williams, Stewards, Prophets, Keepers of the Word: Leadership in the Early Church, reviewed by Joan C. Campbell Arie de Reuver, Sweet Communion: Trajectories of Spirituality from the Middle Ages through the Further Reformation, reviewed by Peter Wyatt Christopher H. Evans, The Kingdom Is Always but Coming: A Life of Walter Rauschenbusch, reviewed by Phyllis Airhart Richard A. Horsley, ed., Christian Origins, reviewed by David A. Reed W.J. Torrance Kirby, Richard Hooker, Reformer and Platonist, reviewed by David Neelands Carolyn Osiek, Margaret Y. MacDonald, with Janet H. Tulloch, A Woman's Place: House Churches in Earliest Christianity, reviewed by Mona Tokarek LaFosse Douglas A. Sweeney, The American Evangelical Story: A History of the Movement, reviewed by Mark D. Chapman Catherine Thom, Early Irish Monasticism: An Understanding of Its. Cultural Roots, reviewed by Thomas P. Power Jiwu Wang, "His Dominion" and the "Yellow Peril": Protestant Missions to Chinese Immigrants in Canada, 1859-1967, reviewed by Ruth Compton Brouwer Ellen B. Aitken, Loosening the Roots of Compassion: Meditations for Holy Week and Eastertide, reviewed by Kathleen a O'Neill Diogenes Allen, Traces of God: 25th Anniversary Edition, reviewed by Mario O. D'Souza Michael A. Hayes, ed., New Religious Movements in the Catholic Church., reviewed by Mario O. D'Souza William F. Haynes, Jr., and Geffrey B. Kelly. Is There a God in Health Care?: Toward a New Spirituality of Medicine., reviewed by Marilyn-Ann Elphick Michael W. Higgins, Stalking the Holy: The Pursuit of Saint Making., reviewed by Tony W. Tremblett Bradford Hinze, Practices of Dialogue in the Roman Catholic Church: Aims and Obstacles, Lessons and Laments, reviewed by Reid B. Locklin Michael Pasquarello III, Christian Preaching: A Trinitarian Theology of Proclamation, reviewed by Mark L. Yensen Robert Stephen Reid., The Four Voices of Preaching: ConnectingPurpose and Identity behind the Pulpit, reviewed by Kerri A. Hagerman Norman Solomon, Richard Harries, Tim Winters, eds., Abraham s Children: Jews, Christians and Muslims in Conversation, reviewed by Mario O. D'Souza James W. Thompson, Pastoral Ministry according to Paul: A Biblical Vision, reviewed by Luke A. Powery Tim P. Van Duivendyk, The Unwanted Gift of Grief A Ministry Approach, reviewed by Colleen Lashmar Halbert Weidner, Grief, Loss and Death: The Shadow Side of Ministry, reviewed by Kathleen O'Neill David Welton, The Treatment of Bipolar Disorder in Pastoral Counseling: Community and Silence, reviewed by Joseph G. Schner Jonathan R. Wilson.,Why Church Matters: Worship, Ministry, and Mission in Practice, reviewed by Joni S. Sancken Todd Bolen, ed., Survey of Western Palestine: The Maps, reviewed by Brian P. Irwin Karen Baker-Fletcher, Dancing with God: The Trinity from a Womanist Perspective, reviewed by Don Schweitzer Michael Mack, Sidney's Poetics: Imitating Creation, reviewed by Meredith Donaldson Clark

Antibacterial SAR for a series of macrolides derived from erythromycin A that are potent LHRH antagonists was developed in an attempt to eliminate the antibiotic activities of these compounds. Increasing the size of the alkyl substituents on the desosamine 3′-amine resulted in potent LHRH antagonists that were inactive against staphylococcal bacteria strains, and were significantly (>10-fold) less active against streptococcal bacteria strains. Complete elimination of antibacterial activities could be achieved by replacement of one or both methyl groups on the 3′-amine with a large alkyl substituent.

The design and synthesis of a series of 11,12-cyclic carbamate derivatives of 6-O-methylerythromycin A that are novel, nonpeptide LHRH antagonists, is described. The macrolide antagonist 1, discovered during a screen of our chemical repository, was compared to a macrocyclic peptide antagonist 2 using molecular modeling, thus providing a model for the design of more potent antagonists. Medicinal chemistry efforts to find a replacement for cladinose at position 3 of the erythronolide core provided a series of oxazolidinone carbamates that were equally as active as the cladinose-containing parent macrolides. The descladinose LHRH antagonist 14 has 1-2 nM affinity for both rat and human LHRH receptors and is a potent inhibitor of LH release (pA2 = 8.76) in vitro. In vivo, 14 was found to produce a dose-dependent suppression of LH in male castrate rats via both i.v. and p.o. dosing.

The case presented is that of a man in his late 60s who experienced an acute anterior myocardial infarction. A complete 2D echocardiogram with color flow and Doppler imaging had been performed on the patient's first hospital day. On day 2 of his hospitalization, a follow-up limited study was performed to reevaluate left ventricular function. Cardiac function was essentially unchanged from the prior day's study. However, color flow Doppler was performed, and a distal septal ventricular septal defect was detected incidentally, only in the final apical view while using the full-height color sector box. This study demonstrates the value of using the full-size color sector box as a screening tool when performing color flow Doppler. Current high-capacity (256 channel and above) equipment assists the sonographer with image optimization and decreases the need to use the truncated color flow box.

Wood polymer composites (WPC) were made from basswood and yellow birch using six cell lumen type polymer formulations. The study was designed to get insight into the influence of wood density and polymer formulation on certain WPC mechanical properties. Small specimens were tested for toughness, stiffness, hardness and bending strength using standard ASTM methods. Results showed that stronger and stiffer polymers produce tougher and stronger WPC, but the effect is small. Thus, there is a wide range of polymer properties which produce useful WPC properties. Study of the fracture surfaces using Scanning Electron Microscopy (SEM) showed that WPC made with different polymers fractured differently and polymer containing a coupling agent bonded to the cell wall. However the cell wall bonding had no noticeable influence on WPC mechanical properties.

Here we describe the three-dimensional crystal structures of human glucocorticoid receptor ligand-binding domain (GR-LBD) in complex with the antagonist RU-486 at 2.3 A resolution and with the agonist dexamethasone ligand together with a coactivator peptide at 2.8 A. The RU-486 structure was solved in several different crystal forms, two with helix 12 intact (GR1 and GR3) and one with a protease-digested C terminus (GR2). In GR1, part of helix 12 is in a position that covers the co-activator pocket, whereas in the GR3, domain swapping is seen between the crystallographically identical subunits in the GR dimer. An arm consisting of the end of helix 11 and beyond stretches out from one molecule, and helix 12 binds to the other LBD, partly blocking the coactivator pocket of that molecule. This type of GR-LBD dimer has not been described before but might be an artifact from crystallization. Furthermore, the subunits of the GR3 dimers are covalently connected via a disulfide bond between the Cys-736 residues in the two molecules. All three RU-486 GR-LBD structures show that GR has a very flexible region between the end of helix 11 and the end of helix 12.

We expressed a test system of wild-type (WT) rat (r) and human (h) gonadotropin-releasing hormone (GnRH) receptors (GnRHRs), including naturally occurring (13) and manufactured (five) "loss-of-function" mutants of the GnRHR. These were used to assess the ability of different GnRH peptidomimetics to rescue defective GnRHR mutants and determine their effect on the level of membrane expression of the WT receptors. Among the manufactured mutants were the shortest rGnRHR C-terminal truncation mutant that resulted in receptor loss-of-function (des(325-327)-rGnRHR), two nonfunctional deletion mutants (des(237-241)-rGnRHR and des(260-265)-rGnRHR), two nonfunctional Cys mutants (C(229)A-rGnRHR and C(278)A-rGnRHR); the naturally occurring mutants included all 13 full-length GnRHR point mutations reported to date that result in full or partial human hypogonadotropic hypogonadism. The 10 peptidomimetics assessed as potential rescue molecules ("pharmacoperones") are from three differing chemical pedigrees (indoles, quinolones, and erythromycin-derived macrolides) and were originally developed as GnRH peptidomimetic antagonists. These structures were selected for this study because of their predicted ability to permeate the cell membrane and interact with a defined affinity with the GnRH receptor. All peptidomimetics studied with an IC(50) value (for hGnRHR)

Combinatorial and structure-based medicinal chemistry strategies were used together to advance a lead compound with an activity of K(i) = 58 microM via a potency enhancement of70 000-fold to an analogue with an activity of K(i) = 0.8 nM against influenza neuraminidase (A/Tokyo/67). Lead optimization was initiated using molecular modeling and combinatorial chemistry. Protein crystal structures revealed that inconsistent structure-activity relationship (SAR) data resulted from different binding orientations of the inhibitor core five-membered rings from one series to another. Binding modes for a series of compounds showed up to a 180 degrees variation in orientation of the five-membered ring within the active site. Potent analogues were only achieved with chemical series that were observed to bind in the same orientation and yielded consistent SAR. In one series, consistent binding was obtained by an unprecedented occupation of a negatively charged binding pocket by a neutral methyl ester unit. The structural rationale for this novel SAR variation, based on protein crystallographic data, is given.

Due to ongoing economic and technological changes over the last two decades, partnerships have become pervasive in tourism development as they have been seen by national governments as one means of dealing with an increasingly complex and multifacted tourist industry. On the island of Ireland, partnership has been established on a trans-jurisdictional basis in an effort to maximise tourist potential and reduce duplication. However, despite this development, very little is known about trans-jurisdictional co-operation as most research to date has focused on local and regional partnerships, and the extent of community participation vis-a-vis administrative authorities. This paper aims to fill the gap in this research by examining the relationship between the two tourist boards in Ireland, the Northern Ireland Tourist Board (NITB) and Bord Failte (BF). By reflecting on the future of cross-border co-operation in tourism on the island of Ireland, the paper outlines important lessons for developing tourism partnerships across national, regional and local administrative boundaries. The paper concludes by arguing that co-operation can be maintained by formulating an inclusive and integrated tourism strategy, establishing partnership balance, understanding political sensitivities and developing a participative partnership approach at the local level.

Although partnerships are emerging as a central feature in governance arrangements in advanced capitalist societies, much discussion has taken place on the value and effectiveness of the approach. In recent years this debate has come to the fore in Northern Ireland, as a plethora of partnerships have been increasingly applied across a wide range of different government programmes at different levels of governance. Using a map of partnership arrangements in Northern Ireland, the author examines three case studies with a view to analysing the value and importance of partnership and assessing the activities that they do best. From this analysis, the author then discusses the future of partnership governance, outlines a number of areas for new research, and presents an agenda for development. As the case studies suggest that too much emphasis has been placed on developing a ‘process’, to the detriment of product outcomes, it is argued that partnerships need to adopt a dual approach to development by combining an inclusive process with greater strategic focus. At the regional level, however, a partnership framework is needed to facilitate coordination, reduce duplication, and improve understanding between partnerships, and between partnerships, local authorities, and government departments. Moreover, by adopting an inclusive approach, a partnership framework has the potential to provide a voice for the socially and politically excluded and to encourage participative planning and pluralist policymaking.

Gonadotropin-releasing hormone (GnRH) stimulates the synthesis and secretion of the gonadotropins that maintain the reproductive axis in mammals. Efforts have focused on the characterization of novel, nonpeptidic orally active antagonists of the GnRH receptor. An erythromycin A derivative, A-198401 (11-deoxy-11-[carboxy (3,4-dichlorophenethyl) amino]-3-O-[4-(S)-methyl-oxazolidin-2-one] carbamoyl-5-O-(3′-N-desmethyl-3′-N-cyclopropylmethyl) desosaminyl-6-O-methyl-erythronolide A 11,12-(cyclic carbamate), showed nanomolar affinity for the human (CHO-21) and rat GnRH receptors in vitro (pK values 8.7 ± 0.2 and 9.2 ± 0.14, respectively). In a functional in vitro assay, A-198401 inhibited leuprolide-induced release of luteinizing hormone (LH) from cultured rat pituitary cells with a pA2 value of 8.8. Intravenous (IV) dosing of A-198401 in castrate male rats produced a significant dose-dependent suppression of LH production with an ED80 value of 5.26 mg/kg. Sustained testosterone (T) suppression was observed after IV dosing of A-198401 in the intact rat, with a 10-mg/kg-dose producing 9–24 h suppression. Analysis of the IV and PO data indicate that A-198401 has a bioavailability of 15%. A-198401 is a novel nonpeptide GnRH antagonist that produces significant and sustained suppression of LH and T production in animal models when dosed either IV or PO and may provide the basis for a therapeutic GnRH antagonist for the clinical treatment of reproductive hormone-dependent diseases. Drug Dev. Res. 52:485–491, 2001. © 2001 Wiley-Liss, Inc.

To increase the efficiency of diffraction data collection for protein crystallographic studies, an automated system designed to store frozen protein crystals, mount them sequentially, align them to the X-ray beam, collect complete data sets, and return the crystals to storage has been developed. Advances in X-ray data collection technology including more brilliant X-ray sources, improved focusing optics, and faster-readout detectors have reduced diffraction data acquisition times from days to hours at a typical protein crystallography laboratory [1, 2]. In addition, the number of high-brilliance synchrotron X-ray beam lines dedicated to macromolecular crystallography has increased significantly, and data collection times at these facilities can be routinely less than an hour per crystal. Because the number of protein crystals that may be collected in a 24 hr period has substantially increased, unattended X-ray data acquisition, including automated crystal mounting and alignment, is a desirable goal for protein crystallography. The ability to complete X-ray data collection more efficiently should impact a number of fields, including the emerging structural genomics field [3], structure-directed drug design, and the newly developed screening by X-ray crystallography [4], as well as small molecule applications.

This paper analyses recent policy developments in civil procurement by UK central government departments and agencies using policy network analysis. The research was funded by an ESRC personal research grant (No. 000237023), and included a questionnaire survey and follow-up interviews of heads of procurement in government departments and agencies, as well as interviews with members of management boards responsible for procurement. The paper maps changes in the policy networks from 1995 to 2000, showing an increasing recognition of the importance of procurement in government policy and a commensurate rise in the status of the procurement function. In particular, the extent to which procurement is seen as contributing to core government policies and processes, such as obtaining better value for money from public expenditure, is reflected in the integration of procurement networks at the centre of government with traditionally powerful policy networks based within Treasury and the Cabinet Office, demonstrated by the establishment of the Office of Government Commerce (OGC) within Treasury. However, that integration has not occurred within all departments and agencies, with the result that procurement's contribution is not being optimised fully across government, and attempts by the centre to ensure uniform implementation of procurement policy raise issues of centre departmental power and interdepartmental relationships. Finally, the paper concludes that policy networks provide a valid and useful theoretical framework for the analysis of public procurement.

The need to decrease the time scale for clinical compound discovery has led to innovations at several stages in the process, including genomics/proteomics for target identification, ultrahigh-throughput screening for lead identification, and structure-based drug design and combinatorial chemistry for lead optimization. A critical juncture in the process is the identification of a proper lead compound, because a poor choice may generate costly difficulties at later stages. Lead compounds are commonly identified from high-throughput screens of large compound libraries, derived from known substrates/inhibitors, or identified in computational prescreeusing X-ray crystal structures. Structural information is often consulted to efficiently optimize leads, but under the current paradigm, such data require preidentification and confirmation of compound binding. Here, we describe a new X-ray crystallography-driven screening technique that combines the steps of lead identification, structural assessment, and optimization. The method is rapid, efficient, and high-throughput, and it results in detailed crystallographic structure information. The utility of the method is demonstrated in the discovery and optimization of a new orally available class of urokinase inhibitors for the treatment of cancer.

Advances in computational methodology and increased structural information about receptors have increased our ability to model and predict ligand-receptor interactions. This review summarizes current computational methods used to develop models of ligand-receptor complexes and new methods that can use these structural models to discover novel ligands.

Renin, the first enzyme in the renin-angiotensin system, is critically important for the maintenance of blood pressure, and, therefore, as a target for antihypertensive therapy. The three-dimensional structure of renin would be an invaluable aid in understanding the functional properties of renin as well as in the design of novel, potent inhibitors. Three-dimensional models of renin have been developed by a number of different groups based on comparative homology modeling from the other known aspartic proteinase structures. These models have been used widely in the drug design process to suggest targets for synthesis and to rationalize the structure-activity relationships of compounds. This review describes the different published renin models and compares them to the extent possible. Applications of these model renin and renin-inhibitor complex structures to biological function and inhibitor design are summarized.

Comparative modeling methods are described that can be used to construct a three-dimensional model structure of a new protein from knowledge of its sequence and of the experimental structures and sequences of other members of its homology family. The methods are illustrated with the mammalian serine protease family, for which seven experimental structures have been reported in the literature, and the sequences for over 35 different protein members of the family are available. The strategy for modeling these proteins is presented, and criteria are developed for determining and assigning the reliability of the modeled structure. Criteria are described that are specially designed to help detect cases in which it is likely that the local structure diverges significantly from the usual conformation of the family.

All reported GnRH receptor mutants (causing human hypogonadotropic hypogonadism) are misfolded proteins that cannot traffic to the plasma membrane. Pharmacoperones correct mis-folding and rescue mutants, routing them to the plasma membrane where they regain function. Because pharmacoperones are often peptidomimetic antagonists, these must be removed for receptor function after rescue; in vivo this necessitates pulsatile pharmacoperone administration. As an antecedent to in vivo studies, we determined whether pharmacoperones need to be present at the time of synthesis or whether previously misfolded proteins could be refolded and rescued. Accordingly, we blocked either protein synthesis or intra-cellular transport. Biochemical and morphological studies using 12 mutants and 10 pharmacoperones representing three different chemical classes show that previously synthesized mutant proteins, retained by the quality control system (QCS), are rescued by pharmacoperones, showing that pharmacoperone administration in vivo likely need not consider whether the target protein is being synthesized at the time of drug administration.

Fortuna Haviv, Wesley Dwight, Bradley Crawford, Rolf Swenson, Milan Bruncko, Michele Kaminski, Kaneyoshi Kato, Yoshihiro Sugiura, Lisa Frey, Gilbert Diaz, Gary Bammert, Eugene N. Bush, Leslie Besecke, Kurt Mohning, Jason Segreti, Mary Spangler, Craig Wegner, and Jonathan Greer Pharmaceutical Products Division, Abbott Laboratories, Abbott Park, IL 60064-3500, U.S.A.; Pharmaceutical Development Division, Takeda Chemical Industries, Osaka Japan; and TAP Pharmaceutical Inc., Abbott Park, IL 60064-3500, U.S.A.

Administration of effective pharmaceutical agents is among the most efficacious and cost-effective ways of treating disease. Therefore, the discovery of new pharmaceutical agents to improve existing therapy and to introduce treatments for previously untreatable conditions is of paramount importance, especially in the current environment of managed health care and the concerted drive to decrease the ever increasing cost of health care. To achieve this goal of continued and even accelerated discovery of new and effective drugs, the drug discovery paradigm has had to evolve in major ways over the past decade or two and is continuing to do so with the discovery and implementation of a broad range of new and advanced technologies.

A novel series of octapeptide LHRH antagonists was designed on the basis of the structure of the (2-9) fragment of a LHRH agonist. By adopting a systematic SAR study, we were able to improve first the in vitro activity and then the in vivo LH suppression, raising them up to the range of the decapeptide antagonists NalGlu (51) and A-75998 (50), resulting in A-76154 (49). The octapeptide antagonist A-76154 is the most potent reduced-size LHRH antagonist reported. It suppresses LH in the castrated rat by over 80% for a period of 4 h following sc bolus administration of 30 micrograms/kg.

Mutagenesis throughout the single-chain urokinase-type plasminogen activator (scu-PA) cDNA molecule, followed by expression of the mutant genes and secretion of the resulting mutant proteins from yeast, has been used to determine the amino acid residues important for activity of scu-PA molecules. Twelve out of 13 colonies secreting variant scu-PA molecules with decreased ability to form a zone of fibrinolysis had mutant genes with a single codon alteration in the serine protease encoding domain (B-chain). Many of these changes are of highly conserved residues in the serine proteases and are consequently of considerable interest. A model three-dimensional structure of the protease domain of urokinase was used to explain the basis for the effects of these down mutations. The model showed that the strongest down mutations result from either interference of the mutated side chain with substrate binding at the active site or the introduction of bulky or charged groups at structurally sensitive internal positions in the molecule. Attempts to find second site revertants of five down mutants, altered either at the plasmin activation site or near the serine at the active site, only resulted in same-site revertants, with the original or closely related amino acids restored.

The ratio of tensile to compressive modulus of elasticity for untreated basswood was approx. 1.9 and for untreated sugar maple approx. 1.3. It approached 1.1 with high polymer loadings. The method of transformed sections gave good estimates of along the grain, tensile, compressive and bending elastic values for maple wood and maple wood polymer composite.