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US 6210874 B1 Abstract wherein x represents an amino acid sequence from position 585 to 604 (gp41-HIV-1) with at least one of a lysine at position 586 or a lysine at both positions 585 and 586; x2 represents an amino acid sequence from position 585 to 604 (gp41-HIV-1); y represents an amino acid sequence from position 612 to 629 (gp41-HIV-1); e and f represent one or more epitopes included in the amino acid sequence extending from 586 to 629 (gp41-HIV-1) or from 578 to 613 (gp36-HIV-2); and a and b represent the amino and carboxy terminals, respectively, as well as substituents effective to make the peptide more useful as an immunodiagnostic. Peptides of formula III have one or both of e and f present. Also provided are peptides of the general formulae:
wherein x1 represents an amino acid sequence from position 577 to 596 (gp36-HIV-2) with at least one of a lysine at position 578 or a lysine at both positions 577 and 578; x3 represents an amino acid sequence from position 577 to 596 (gp36-HIV-2); y1 represents an amino acid sequence from position 604 to 613 (gp36-HIV-2); e and f represents one or more epitopes included in the amino acid sequence from 578 to 613 (gp36-HIV-2) or 586 to 629 (gp41-HIV-1); and a and b are as defined above. Peptides of formula IV have one or both of e and f present. These peptides are useful in detecting HIV-1 and HIV-2 antibodies.
Claims(3) wherein:
a represents the H group which attaches to form the amino terminus or a substituent effective as a coupling agent to make the peptide more useful as an immunodiagnostic reagent without changing its antigenic properties;
b represents the OH group which attaches to form the carboxy terminus or a substituent effective as a coupling agent to make the peptide more useful as an immunodiagnostic reagent without changing its antigenic properties;
x is KILAVERYLKDQQLLGIWG;
y is TTAVPWNA; and
f is SGKLI.
a represents the H group which attaches to form the amino terminus or a substituent effective as a coupling agent to make the peptide more useful as an immunodiagnostic reagent without changing its antigenic properties; and
b represents the OH group which attaches to form the carboxy terminus or a substituent effective as a coupling agent to make the peptide more useful as an immunodiagnostic reagent without changing its antigenic properties.
3. A method for detecting the presence of antibodies to HIV-1, said method comprising contacting an analyte suspected of containing said antibodies with the peptide of claim 1 in a manner and for a time sufficient to allow binding of said antibodies to said peptide, and detecting binding of said antibodies to said peptide.
Description This is a continuation of application Ser. No. 08/077,085, filed Jun. 16, 1993, now abandoned, which is a division of application Ser. No. 07/549,964, filed Jul. 9, 1990, now U.S. Pat. No. 5,241,047; which is a CIP of application Ser. No. 07/281,205, filed Dec. 8, 1988, which is a CIP of application Ser. No. 07/185,518, filed Apr. 22, 1988, now abandoned; which is a CIP of application Ser. No. 07/148,821, filed Jan. 27, 1988. The present invention relates to novel cyclic peptides and combinations thereof alone and with linear and cyclic peptides for detecting HIV antibodies. Acquired Immune Deficiency Syndrome (AIDS), AIDS related complex (ARC) and pre-AIDS are thought to be caused by a retrovirus, the Human Immunodeficiency Virus (HIV). The first AIDS related virus, HIV-1 (also known as HTLV-III, LAV-1 and ARV) has been well characterized. Another pathogenic human retrovirus named HIV-2 (formerly LAV-2) has now been isolated from West African patients with AIDS. See, e.g., WO 87/04459. HIV-2 has recently been shown (Guyader et al. Nature 326 662-669, 1987) to share a number of conserved sequences with HIV-1 and the Simian Immunodeficiency Viruses (SIV). Even though other numbering systems are used in the art, for ease of understanding and comparison we have adopted herein the amino acid numbering system of Ratner et al., Nature, 313, 277-284, 1985 for the HIV-1 proteins and that of Guyader et al., Nature 326, 662-669 (1987) for the HIV-2 proteins. The amino acids in the peptides of this invention are designated by the single letter code as follows: ala=A, arg=R, asn=N, asp=D, cys=C, gln=Q, glu=E, gly=G, his=H, ile=I, leu=L, lys=K, met=M, phe=F, pro=P, ser=S, thr=T, trp=W, tyr=Y and val=V. The initial immunodiagnostic tests for the detection of antibodies in the serum of patients infected with HIV-1 utilized the whole virus as antigen. Second generation tests made use of polypeptide sequences obtained by the recombinant DNA methodology. Cabradilla et al. Bio/Technology 4 128-133 (1985) and Chang et al. Bio/Technology 3, 905-909 (1985), for example, refer to bacterially synthesized viral protein fragments of 82 and 102 amino acid residues, respectively. EPA 202314 and 114243 refer to recombinant polypeptides spanning regions of gp41 and gp120 that are immunoreactive alone or in mixtures. Shoeman et al., Anal. Biochem. 161, 370-379 (1987) refers to several polypeptides from gp41 that are immunoreactive with antibodies present in sera from patients infected with HIV-1. None of the above assay procedures is, however, totally acceptable. Their lack of sensitivity is a critical shortcoming. It may permit blood containing virus to escape detection and thereby potentially result in the infection of blood product receivers and continued infectivity by undiagnosed AIDS carriers. Their lack of specificity (false positives) is also a problem�healthy individuals are told they may have AIDS. Such false positive may be caused by impurities. They may also be caused by shared epitopes with viruses unrelated to AIDS present in these antigen preparations. In this regard, Gallaher, Cell 50 327-328, 1987 has reported that a region of gp41 of HIV-1 shares a sequence of five adjacent amino acid residues with the respiratory syncytial virus and of four equally distributed amino acids of the measles virus F1 glycoprotein. Thus, even highly purified recombinant polypeptides containing this region, or any other common regions yet to be discovered, could potentially be responsible for false positives and the attendant unacceptable specificity. Finally, these prior art assays do not permit detection of very low levels of HIV antibodies. This disadvantages the assays in terms of their ability to detect AIDS infections at a very early stage, thereby delaying the start of treatment and permitting the possible spread of infection by blood samples and other body fluids before effective detection of AIDS infection. In an attempt to solve these problems, diagnostic means and methods employing shorter HIV antigens are now being developed. Empirical methods to identify peptide sequences corresponding to unique and highly conserved epitopes of the HIV viruses are also now available. These methods are, for example, capable of assisting in the selection of short amino acid sequences which are more likely to be exposed on the surface of the native protein and thus useful as assay tools (for a review see Hopp and Woods, J. Immunol. Met. 88, 1-18, 1986). Although somewhat useful, these methods are no more than indicative. Nonetheless, they have been applied to identify epitopes present on the surface of viruses responsible for AIDS. For example, U.S. Pat. No. 4,629,783, International Patent Appl. PCT/US86/00831 and EPA 303224 refer to various synthetic peptides from the p18, p25, gp41 and gp120 proteins. These peptides are advantaged by the relative ease and lower cost with which they can be prepared and more importantly because of the reduced risk of obtaining false positives with them due to impurities or the presence of shared epitopes with viral proteins not related to AIDS. While these smaller peptides are advantaged in terms of specificity over the earlier recombinant polypeptide and whole virus approaches to the diagnosis of AIDS infections, they have been less than satisfactory in terms of overall sensitivity, perhaps because the synthesized epitope is not able to assume and maintain a conformation that is recognized by the AIDS antibodies. Although the number of serum samples tested in each of these cases is very limited, specificity (few if any false positives) was found to be very high (95%-100%) with the small synthetic peptides but the overall sensitivity varied between 80% and 100%. In fact, in the only example where 100% sensitivity was attained only ten samples were tested. For example, Smith et al., J. Clin. Microbiol. 25 1498-1504, 1987 refers to two overlapping peptides, E32 and E34, that are highly immunoreactive. No false positives, out of 240 seronegative specimens, were obtained but the peptides missed three seropositive samples out of 322 (sensitivity of 99.1%). Wang et al. (Proc. Natl. Acad. Sci 83, 6159-6163, 1986) refers to a series of overlapping peptides (including amino acid residues of Smith's E32 and E34 peptides) among which one 21-mer peptide showed 100% specificity and 98% sensitivity (out of 228 seropositive samples taken from patients with AIDS, 224 were found positive with this peptide). And U.S. patent application 120,027, filed Nov. 13, 1987 refers to a short synthetic peptide spanning residues 606 to 620 (SGKLICTTAVPWNAS) of gp41 (HIV-1). This peptide is said to be immunoreactive with antibodies of patients infected by the AIDS viruses. The specificity was also excellent (63/63) but 6 seropositive specimens out of 57 confirmed positive could not be detected (sensitivity of 89%). Gnann et al. (J. Virol. 61, 2639-2641, 1987 and J. Infec. Dis 156, 261-267, 1987) also refer to a series of overlapping peptides from a suspected immunodominant region of gp41 (HIV-1). Gnann et al. concluded that cys-605 was essential for the immunoreactivity of that segment of the gp41-(HIV-1) protein. They reported that a peptide having the sequence SGKLIC (606-611) was not immunoreactive with any of the 22 HIV-1 positive sera tested, while the addition of the cysteine residue to the N-terminus restored some immunoreactivity, 21 of 44 sera reacted with the 7-mer peptide (48% sensitivity). Gnann et al. (J. Virol) also speculated that the cysteine residues at positions 605 and 611 of gp41 (HIV-1) might play a role in the antigenic conformation of this region perhaps by the formation of a cyclic structure via disulfide bonding. However, Gnann et al. never demonstrated that they did have a synthetic peptide wherein the two cysteine groups were linked by disulfide bonds. Although Gnann et. al refers to peptides which are useful in identifying HIV-1 antibodies, even its peptides lack 100% sensitivity. For example, Gnann et al. (J. Virol. 61, 2639-2641, (1987)) report that while their 600-611 amino acid sequence detected 22 out of 22 positive sera, they also reported that similar tests carried out at the Centers for Disease Control, Atlanta, Ga. with the same 12-amino acid sequence (600-611) missed 1 out of 79 positive sera. And Gnann et al. in J. Infect. Dis. 156, 261-267, 1987 reported that the same 12-amino acid sequence was reactive with 131 out of 132 HIV-1 infected patients from the United States. Gnann et al. Science 237, 1346-1349, 1987 reports a short linear synthetic peptide spanning residues 592 to 603 of gp42 (HIV-2) that contains two cysteines in a region homologous to the 605-611 region of gp41(HIV-1). This peptide reacted with 5 out of 5 sera taken from HIV-2 infected patients. Other peptides containing amino acids 605-611 of gp41 of (HIV-1) are also referred to in the art. WO 86/06414 refers to peptide X(39), which is encoded by the region from about bp 7516 through bp 7593, and peptide XIII(79) which is encoded by the region extending from about bp 7543 through bp 7593, both containing the 7-amino acid sequence 605-611. These peptides are reported to be linear and no formation of cyclic structures is suggested. WO 87/06005 reports that a series of synthetic peptides encompassing the Cys(605)-Cys(611) residues of the HIV-1 envelope glycoprotein (gp41) undergo a series of spontaneous oxidative transformations upon solubilization in neutral or basic aqueous buffer. It speculates that as a result, the peptides when used in ELISAs are a random mixture of linear monomer, cyclic monomer, linear or cyclic dimers and linear polymers of various lengths. The application did not actualy demonstrate the presence of cyclic components and did not characterize the other various dimers and polymers possibly present. Moreover, it speculates that the polymer forms are the most important components for ELISA reactivity. In addition to perhaps being complex mixtures of various oxidative forms of the peptide, the prior art peptides referred to above do not permit as early detection of AIDS infection as would be desirable. For example, Gnann et al. (J. Virol) reports that when the HIV-1 positive sera are diluted by a factor exceeding 500, some of these diluted sera are found to be negative thus indicating a low sensitivity of the peptide for early HIV detection. These problems have been addressed by employing peptides that have been chemically cyclized to form a disulfide bridge between the relevant cysteines. E.g., M. Lacroix et al., Comparative Performance of Cyclic Versus Linear Peptides In An ELISA For HIV-1 And HIV-2 Specific Antibodies, No. 3147, June 1989 AIDS Conference, Montreal, Canada; and WO 89/03344. This invention is directed to improvements in such cyclic peptides. In accordance with the present invention, there is provided a novel series of peptides which are particularly adapted for detecting 100% of HIV-1 and HIV-2 antibodies and which are capable of detecting such antibodies even when present in very low levels in sera. More specifically, the novel peptides of the present invention are selected from substantially pure peptides of formulae I or II: wherein: x is independently selected from one of the following amino acid sequence analogs of the amino acid sequence of gp41-HIV-1: KILAVERYLKDQQLLGIWG- (586-604) KKILAVERYLKDQQLLGIWG- (585-604), amino acid sequences corresponding thereto, which sequences are derived from homologous regions of other HIV-1 isolates, and amino acid sequences differing from the above as a result of conservative substitutions, such amino acid sequences being characterized by at least one of a lysine at position 586 or a lysine at both positions 585 and 586; y, if present, is independently selected from the group consisting of: amino acid sequences corresponding thereto, which sequences are derived from homologous regions of other HIV-1 isolates, and amino acid sequences differing from the above as a result of conservative substitutions; e and f, if present, are independently selected from the group consisting of an amino acid sequence of any one of the epitopes of the region spanning amino acids 586 to 629 of gp41 of HIV-1 or of the region spanning amino acid sequence 578 to 613 of gp36 of HIV-2, amino acid sequences corresponding thereto and being derived from homologous regions of other HIV-1 or HIV-2 isolates, amino acid sequences differing from the above as a result of conservative substitutions, and any combination of these epitopes; a is an amino terminus or a substitutent effective as a coupling agent and/or to make the peptide more useful as an immunodiagnostic reagent without changing its antigenic properties; and b is a carboxy terminus or a substituent effective as a coupling agent and/or to make the peptide more useful as an immunodiagnostic reagent without changing its antigenic properties; and wherein: x1 is independently selected from one of the following amino acid sequence analogs of the amino acid sequence of gp36-HIV-2: KVTAIEKYLQDQARLNSWG (578-596) KKVTAIEKYLQDQARLNSWG (577-596), amino acid sequences corresponding thereto, which sequences are derived from homologous regions of other HIV-2 isolates and amino acid sequences differing from the above as a result of conservative substitutions, such sequences being characterized by at least one of a lysine at position 578 or a lysine at both positions 577 and 578; y1, if present, is independently selected from the group consisting of: amino acid sequences corresponding thereto, which sequences are derived from homologous regions of other HIV-2 isolates, and amino acid sequences differing from the above as a result of conservative substitutes; and e, f, a and b are as defined above. In another embodiment, the novel peptides of this invention are selected from substantially pure peptides of the formulae III and IV: wherein x2, if present, is independently selected from the group consisting of: amino acid sequences corresponding thereto, which sequences are derived from homologous regions of other HIV-1 isolates, and amino acid sequences differing from the above as a result of conservative substitutions; and y, e, f, a and b are as previously defined, one or both of e or f being present; and wherein x3, if present, is independently selected from the group consisting of: amino acid sequences corresponding thereto, which sequences are derived from homologous regions of other HIV-2 isolates, and amino acid sequences differing from the above as a result of conservative substitutions; and y1, e, f, a and b are as previously defined, one or both of e or f being present. A particularly preferred peptide of formula I is BCH-408 which has the following sequence: This peptide incorporates at the f position 619 the amino acid sequence of the epitope located at position 606-610 (SGKLI) of gp41 of HIV-1 and at position 586 a lysine. A particularly preferred peptide of formula II is BCH-417 which has the following sequence: This peptide incorporates at the f position 613 the amino acid sequence of the epitope located at position 598-602 (AFRQV) of gp36 of HIV-2 and at position 578 lysine. Selection of Peptides for Synthesis The peptides of this invention were synthesized on the basis of published amino acid sequences of HIV-1 and HIV-2. However, it should be understood that sequences derived from the homologous regions of other HIV-1 or HIV-2 isolates can be used without departing from the scope of this invention. Epitopes in these native sequences were chosen for use as e and f, in the peptides of this invention using various physicochemical principles that aid in predicting which portions of a polypeptide are most likely to be surface oriented and therefore immunogenic. These include the hydrophilicity plots of Hopp and Woods (Proc. Natl. Acad. Sci. 78, 3824-3828, 1981), and a similar approach by Kyte and Doolittle (J. Mol. Biol. 157, 105-132, 1982). Also, the empirical prediction of protein conformation (Chou and Fasman, Ann. Rev. Biochem. 47, 251-276, 1978) is a useful guide in predicting which parts of the polypeptide are likely to be immunogenic. The e and f epitopes of the peptides of this invention include, but are not limited to, WGCAF (identified by Norrby et al., AIDS Research and Human Retroviruses, Vol. 5, No. 5, 1989); KD, SGKL, and LEDQ (identified by Norrby et al., AIDS, Vol. 3, No. 1 (1989); LKDQ, CSGKLI, and IWG (identified by Mathiesen et al., Immunology, 67 1-7 (1989); and ARILAVERYLKD, and SGKLICTTAVPWNAS (identified by Dopel et al., Jol. of Vir. Meth., 25 167-178 (1989). It is also within the scope of this invention to modify the peptides of this invention, in order to make them more useful as immunodiagnostic reagents without changing their antigenic properties. Such changes include: addition of a cysteine residue at the amino or carboxy terminus in order to facilitate coupling of the peptide to a carrier protein with heterobifunctional cross-linking reagents such as sulfosuccinimidyl-4(p-maleimidophenyl)butyrate, a preferred reagent for effecting such linkages; addition of certain amino acids at the amino or carboxy terminus to facilitate linking of peptides to each other, for coupling to a support or larger peptide or for modifying the physical or chemical properties of the peptide. Such changes may be effected, for example, by additions of tyrosine, glutamic acid or aspartic acid, which can be used as linkers via an esterification reaction, and lysine which can be connected by Schiff base or amide formation; and derivatization by amino terminal acylation, thioglycolic acid amidation, and carboxy terminal amidation, e.g. using ammonia, methylamine. These modifications result in changes in net charge on the peptide and can also facilitate covalent linking of the peptide to a solid support, a carrier or another peptide. These modifications are not likely to result in immunoreactivity changes to the peptide. The peptides of this invention may also be modified by various changes such as insertions, deletions and substitutions, either conservative or nonconservative where such changes might provide for certain advantages in their use. These changes include preferably the following conservative changes: gly, ala; val, ile, leu; asp, glu; asn, gln; ser, thr; lys, arg; phe, tyr; ala, ser; ala, thr; ala, val; ala, pro; ala, glu; leu, gln; gly, phe; ile, ser; and ile, met. Methionine, an amino acid which is prone to spontaneous oxidation, can also usually be replaced by norleucine without changing antigenicity. It may also be convenient to add a �tail� consisting of a small number (1-10) of hydrophobic amino acids to the peptides of this invention. Such tails may facilitate passive adsorption of a peptide to a solid support. This modification can be made at either the COOH or NH2 termini. The preferred addition is phe-ala-phe-ala-phe. In accordance with this invention, the preferred cyclic peptides of formula I are those having x, y, e and f defined as follows: x: KILAVERYLKDQQLLGIWG, y: TTAVPWNAS, e and f not present (BCH-87ck); x: KKILAVERYLKDQQLLGIWG, y: TTAVPWNAS, e and f not present (BCH-266); and x: KILAVERYLKDQQLLGIWG, y: TTAVPWNA, f: SGKLI and e not present (BCH-408), BCH-408 being the most preferred. The preferred cyclic peptides of formula II are those having x1, y1, e and f defined as follows: x1: KVTAIEKYLQDQARLNSWG, y1: HTTVPWVNDS and e and f not present (BCH-202ck); x1: KKVTAIEKYLQDQARLNSWG, y1: HTTVPWVNDS and e and f not present (BCH-265); and x1: KVTAIEKYLQDQARLNSWG, y1: HTTVPWVNDS and f: AFRQV and e not present (BCH-417), BCH-417 being the most preferred. TABLE I provides the full amino acid sequences of these preferred peptides (disregarding possible a and b):
Preparation of Linear and Cyclic Peptides The peptides of this invention are preferably prepared using conventional solid phase synthesis. However, other well known methods of peptide synthesis may also be used. The resin support is any suitable resin conventionally employed in the art for solid phase preparation of polypeptides, preferably p-benzyloxyalcohol polystyrene and p-methylbenzydrylamine resin. Following the coupling of the first protected amino acid to the resin support, the amino protecting group is removed by standard methods conventionally employed in the art of solid phase peptide synthesis. After removal of the amino protecting group, remaining α-amino protected and, if necessary, side chain protected amino acids are coupled, sequentially, in the desired order to obtain the product. Alternatively, multiple amino acid groups may be coupled using solution methodology prior to coupling with the resin-supported amino acid sequence. The selection of an appropriate coupling reagent follows established art. For instance, suitable coupling reagents are N,N′-diisopropylcarbodiimide or N,N′-dicyclohexylcarbodiimide (DCC) either alone or preferably in the presence of 1-hydroxybenzotriazole. Another useful coupling procedure makes use of preformed symmetrical anhydrides of protected amino acids. The necessary α-amino protecting group employed for each amino acid introduced onto the growing polypeptide chain is preferably 9-fluorenylmethyloxycarbonyl (Fmoc), although any other suitable protecting group may be employed as long as it does not degrade under the coupling conditions and is readily removable selectively in the presence of any other protecting groups already present in the growing molecule. The criteria for selecting protecting groups for the side chain amino acids are: (a) stability of the protecting group to the various reagents under reaction conditions selective for the removal of the α-amino protecting group at each step of the synthesis: (b) retention of the protecting group's strategic properties (i.e. not be split off under coupling conditions) and (c) removability of the protecting group upon conclusion of the polypeptide synthesis and under conditions that do not otherwise affect the polypeptide structure. The fully protected resin-supported peptides are cleaved from the p-benzyloxy alcohol resin with 50 to 60 percent solution of trifluoroacetic acid in methylene chloride for 1 to 6 hours at room temperature in the presence of appropriate scavengers such as anisole, thioanisole, ethyl methyl sulfide, 1,2-ethanedithiol and related reagents. Simultaneously, most acid labile side-chain protecting groups may then be removed. More acid resistant protecting groups are removed by HF treatment. Cyclic peptides of this invention are prepared by the direct oxidative conversion of protected or unprotected SH-groups to a disulfide bond by following techniques generally known in the art of peptide synthesis. The preferred method involves the direct oxidation of free SH-groups with potassium ferricyanide. Such cyclic peptides are believed to assume a more rigid conformation which may favor binding to HIV antibodies. Peptide Mixtures and Polymers Within the scope of this invention are larger peptides formed by the covalently linking of one or more peptides of this invention. Polymers (both homo and co) of these peptides are also envisioned. Also within the scope of the present invention are other cyclic and mixtures of the cyclic peptides of this invention and other cyclic and linear HIV derived peptides. These mixtures have surprisingly been found to provide high sensitivity detection of HIV-1 and HIV-2 antibodies present in serially diluted serum samples and in seroconversion panels (HIV-1). Also it has been found that these mixtures provide a high level of specificity resulting in a minimal number of false positives. Such mixtures comprise at least one cyclic peptide of the general formulae I or III (preferably BCH-87ck, BCH-266 or BCH-408 and more preferably BCH-408) in combination with at least one cyclic peptide of the general formulae II or IV (preferably BCH-202ck, BCH-265 or BCH-417 and more preferably BCH-417). HIV Antibody Detection The peptides and the peptide mixtures of the present invention are useful as diagnostic reagents for the detection of AIDS-associated antibodies in accordance with methods well-known in the art. These include ELISA, hemagglutination, single-dot and multi-dot methods and assays. The main advantage of the present peptides in the determination of antibodies against AIDS resides in their specificity and high sensitivity, and particularly their ability to detect the presence of very low levels of AIDS infection, when compared with known antigens used so far. According to one method for the determination of antibodies against HIV-1 or HIV-2, the so-called �Western Blotting� analysis is used [Towbin, H., Staehelin, T. and Gordon, J., Proc. Nat. Acad. Sci. U.S.A. 76, 4350-4354 (1979)]. According to this technique a peptide or peptides of the present invention is (or are) applied to nitrocellulose paper. The nitrocellulose paper is saturated and then treated with the serum to be tested. After washing, the nitrocellulose paper is treated with an anti-human IgG labeled with an enzyme. The enzymatic activity is then determined by a suitable substrate. Of course, other labels, like radioactive or fluorescence labels, may be used. A preferred convenient and classical technique for the determination of antibodies against HIV-1 or HIV-2 using a peptide or a peptide mixture of the present invention is an enzyme-linked immunosorbent assay (ELISA). In this assay, for example, a peptide, peptide mixture or combination of the present invention is adsorbed onto, or covalently coupled to, the wells of a microtiter plate. The wells are then treated with the sera or analyte to be tested. After washing, anti-human IgG or antihuman IgM labeled with peroxidase is added to the wells. The determination of the peroxidase is performed with a corresponding substrate, e.g., with o-phenylene diamine. Without departing from the usefulness of the illustrative assay, the peroxidase can be exchanged by another label, e.g., by a radioactive, fluorescence chemiluminescence or infra-red emitting label. In the ELISA test, it is possible to use individual peptides or a combination thereof. The latter is preferable since it allows one to combine the most effective peptides for detecting antibodies while at the same time excluding those that contribute to false responses. It was discovered during the course of these studies that some serum samples gave correct positive results with mixtures of peptides while giving equivocal responses with individual peptides as antigen. Thus the most preferred test for HIV-1 and HIV-2 antibodies is achieved in accordance with this invention using a combination of peptide antigens. Another method for the determination of antibodies against HIV-1 or HIV-2 with the peptides or mixture of peptides of this invention is an enzyme immunological test according to the so-called �Double-Antigen-Sandwich-Method�. This method is based on the work of Maiolini as described in Immunological Methods 20, 25-34 (1978). According to this method, the serum or other analyte to be tested is contacted with a solid phase on which a peptide or mixture of peptides of the present invention has been coated (capture layer) and with a peptide or a peptide mixture of the present invention which is labeled with peroxidase (probe layer). The immunological reaction can be performed in one or two steps. If the immunological reaction is performed in two steps, then a washing step is preferably performed between the two incubations. After the immunological reaction or reactions, a washing step may also be performed. Thereafter, the peroxidase is determined with a substrate, e.g., with o-phenylene diamine. Other enzymes and chromogens, including those already described can also be employed in this assay. Suitable solid phases are organic and inorganic polymers, such as amylases, dextrans, natural or modified celluloses, polyethylenes, polystyrenes, polyacrylamides, agaroses, magnetites, porous glass powders, polyvinyldiene fluoride (kynar) and latex, the inner wall of test vessels e.g., test tubes, titer plates or cuvettes of glass or articifial material as well as the surface of solid bodies, e.g., rods of glass and artificial material, rods with terminal thickening, rods with terminal lobes or lamallae. Spheres of glass and artificial material are especially suitable solid phase carriers. The peptides and mixtures of peptides of the present invention are not only useful in the determination of antibodies against HIV-1 or HIV-2, but also indirectly for the determination of HIV-1 or HIV-2 itself since these peptides either free, polymerized or conjugated to an appropriate carrier are useful in eliciting antibodies, in particular monoclonal antibodies, against HIV-1 or HIV-2. Such antibodies can be produced by injecting a mammalian or avian animal with a sufficient amount of a peptide or mixture of peptides of the present invention and recovering said antibodies from the serum of said animals. Suitable host animals for eliciting antibodies include mammals such as rabbits, horses, goats, guinea-pigs, rats, mice, cows, sheep, etc.. Various methods which are generally known can be employed using the peptides of this invention or mixtures thereof in the quantitative determination of HIV-1 or HIV-2 infection. In one such procedure known amounts of a serum sample to be assayed, radiolabeled cyclic peptide of the present invention or mixtures of those peptides and unlabeled peptide or mixture of peptides of the present invention are mixed together and allowed to stand. The antibody/antigen complex is separated from the unbound reagents by procedures known in the art, i.e., by treatment with ammonium sulfate, polyethylene glycol, a second antibody either in excess or bound to an insoluble support, dextran-coated charcoal and the like. The concentration of the labeled peptide or mixture of peptides of the present invention is determined in either the bound or unbound phase and the HIV-1 or HIV-2 content of the sample can then be determined by comparing the level of labeled component observed to a standard curve in a manner known �per se�. Another suitable quantitative method is the �Double-Antibody-Sandwich-Assay�. According to this assay the sample to be tested is treated with two different antibodies raised against a peptide of this invention or mixture thereof using different animals, e.g. sheep or rabbits. Alternatively, monoclonal antibodies may be prepared using the well-known Koehler and Milstein technique for producing monoclonal antibodies. In order to distinguish monoclonal antibodies which are directed against the same antigen, but against different epitopes, the method of St�hli et al. [J. of Immunological Methods 32, 297-304 (1980)] can be used. It is also appropriate to use a polyclonal antiserum and a monoclonal antibody. One of these antibodies is labeled and the other is coated on a solid phase. The suitable solid phases are those mentioned earlier in this application. Suitable labels are enzymes, e.g. peroxidase, radioactive labels or fluorescence-labels. The preferred solid phase is a plastic bead and the preferred label is horse-radish peroxidase. The sera sample is then incubated with the solid phase antibody and the labeled antibody. It is possible to treat the sample first with the solid phase antibody and after washing to treat the sample with the labeled antibody. However, it is also possible to treat the sample first with the solid phase antibody and after a certain time with the labeled antibody. Preferably the sample is treated together with the solid phase and the labeled antibody. After the immunological reaction(s), a washing step may be performed. After washing, the label is determined according to procedures known in the art. In the case where peroxidase is used as the label, the determination is performed with the substrate, e.g., with o-phenylene diamine or with tetramethylbenzidine. The amount of the labeled component is proportional to the amount of the antigen(s) present in the sample. The methods and assays for the determination and quantification of HIV-1, HIV-2 or of antibodies against HIV-1 or HIV-2 as described above can be conducted in suitable test kits comprising, in a container, a cyclic peptide of the present invention, peptide mixtures or a combination thereof, or antibodies against HIV-1 or HIV-2 elicited by a cyclic peptide or a mixture of cyclic and linear peptides of the present invention. Panel of Sera Tested To demonstrate the surprising sensitivity and specificity of the peptides of this invention a panel of sera was tested with illustrative peptides. O.D. Values were obtained at 450 nm and the blank values measured with the sample dilution buffer were not subtracted. Samples NEIA-2*2, BBI-1-162 to 168, 87B140, 87L139, 87V103 are all negative for HIV antibodies. Sample LSPQ-S9-1 is an early seroconverter (HIV-1). The series labeled CAP-113 to CAP-120 corresponds to a pool of seven HIV-1 positive plasma samples serially diluted with an HIV-negative plasma. CAP-113 is the pool diluted 50-fold with the HIV-negative plasma; CAP-114 is diluted 100-fold; CAP-115 is diluted 200-fold; etc. Similarly, the series labeled CAP-222 to CAP-230 corresponds to a pool of seven HIV-2 confirmed seropositive plasma samples. CAP-222 is diluted by 50 with an HIV-negative plasma; CAP-223 by 100; etc. Before the assay is done, each sample, including the CAP-series, is further diluted by 50 with the sample dilution buffer. In these tests, the cut-off for seropositivity is defined as the sum of the O.D. value for sample NEIA-2*2 plus 0.100. Results The cyclic peptides of the present invention were coated and tested in accordance with the ELISA test described previously. TABLE 2 compares the sensitivity of peptide BCH-87c (586(arginine)) to the sensitivity of peptide BCH-87ck (586(lysine)) and peptide BCH 266 (586(lysine)�585(lysine)) at progressively higher dilutions of the antibody in the sera samples. It was found that the substitution of a lysine for the arginine at amino acid position 586 increased the sensitivity in detecting HIV-1 antibodies. A further increase in sensitivity was obtained by an additional lysine at position 585. TABLE 3 compares the activity of BCH-87c, BCH-87ck, BCH-266 and BCH-408 at progressively higher dilutions. It is evident from TABLE 3 that peptide BCH-408, wherein an important epitope located at position 606-610 (SGKLI) is repeated at its c-terminus, has superior sensitivity compared to the other peptides. It was found with the HIV-2 peptides that substituting a lysine for the arginine at amino acid position 578 increased the sensitivity in detecting HIV-2 antibodies. A further increase in sensitivity was obtained by an additional lysine at position 577. TABLE 4 compares the sensitivity of the peptide BCH-202c (578(arginine)) to the sensitivity of peptide BCH-202ck (578(lysine)) and peptide BCH 265 (598(lysine)�577(lysine)) at progressively higher dilutions of the antibody in the sera samples. Peptide cocktails were also made to detect a mixture of HIV-1 and HIV-2 antibodies. TABLE 5 illustrates the sensitivity of peptide cocktail mixtures BCH-87c and BCH-202c (arginine) versus BCH-87ck and BCH-202ck (lysine). The peptide cocktail which included the peptides with lysine substituted for arginine have a higher sensitivity in detecting HIV antibodies.
The following illustrates the general procedures for the synthesis and utilization of the peptides of this invention. The desired Nα-FMOC protected amino acid residue in a mixture of methylene chloride (CH2Cl2) and dimethylformamide (DMF) (4:1) was added to a suspension of p-benzyloxy alcohol resin in CH2Cl2:DMF (4:1) at 0� C. The mixture was stirred manually for a few seconds and then treated with N,N′-dicyclohexylcarbodiimide (DCC) followed by a catalytic amount of 4-(dimethylamino)pyridine. The mixture was stirred at 0� C. for an additional 30 minutes and then at room temperature overnight. The filtered resin was washed successively with CH2Cl2, DMF and isopropanol (3 washes each) and finally, with CH2Cl2. The resin was suspended in CH2Cl2, chilled in an ice bath and redistilled pyridine was added to the stirred suspension Benzoyl chloride was then also added. Stirring was continued at 0� C. for 30 minutes and then at room temperature for 60 minutes. After filtration, the resin was washed successively with CH2Cl2, DMF and isopropanol (3 washes each) and finally with petroleum ether (twice) before being dried under high vacuum to a constant weight. Spectrophotometric determination of substitution according to Meienhofer et al. (Int. J. Peptide Protein Res., 13, 35, 1979) indicates the degree of substitution on the resin. The resin carrying the Nα-FMOC protected first amino acid residue was placed in a reaction vessel of a Biosearch 9600 Peptide Synthesizer and treated as follows: 1) Washed with DMF (4 times for 20 sec. each) 2) Prewashed with a 30% solution of piperidine in DMF (3 min.) 3) Deprotected with a 30% solution of piperidine in DMF (7 min.) 4) Washed with DMF (8 times for 20 sec. each) 5) Checked for free amino groups�Kaiser Test (must be positive) 6) The peptide resin was then gently shaken for 1 or 2 hrs with 8 equivalents of the desired FMOC-protected amino acid and 1-hydroxybenzotriazole and benzotriazol-1-yl-oxy-tris(dimethyl-amino)phosphonium hexafluorophosphate all dissolved in dry redistilled DMF containing 16 equivalents of 4-methylmorpholine. 7) Washed with DMF (6 times for 20 sec. each) After step 7, an aliquot was taken for a ninhydrin test. If the test was negative, one goes back to step 1 for coupling of the next amino acid. If the test was positive or slightly positive, steps 6 and 7 should be repeated. The above scheme maybe used for coupling each of the amino acids of the peptides described in this invention. Nα-protection with FMOC may also be used with each of the remaining amino acids throughout the synthesis. Radiolabeled peptides may be prepared by incorporation of a tritiated amino acid using the above coupling protocol. After the addition of the last amino acid, the Nα-FMOC of the N-terminal residue is removed by going back to steps 1-7 of the above scheme. The peptide resin is washed with CH2Cl2 and dried in vacuo to give the crude protected peptide. The protected peptide-resin was suspended in a 55% solution of trifluoroacetic acid (TFA) in CH2Cl2, containing 2.5% ethanedithiol and 2.5% anisole. The mixture was flushed with N2 and stirred for 1.5 hours at room temperature. The mixture was filtered and the resin washed with CH2Cl2. The resin was treated again with 20% TFA in CH2Cl2 for 5 minutes at room temperature. The mixture was filtered and the resin washed with 20% TFA in CH2Cl2 and then washed with CH2Cl2. The combined filtrates were evaporated in vacuo below 35� C. and the residue triturated several times with dry dimethyl ether. The solid was dissolved in 10% aqueous acetic acid and lyophilized to afford the crude product. The peptides containing arg and cys residues are further deprotected by HF treatment at 0� C. for 1 hour in the presence of anisole and dimethylsulfide. The peptides were extracted with 10% aqueous acetic acid, washed with dimethyl ether and lyophilized to afford the crude peptides. The crude peptides were purified by preparative HPLC on a Vydac column (2.5�25 mm) of C18 or C4 reverse phase with a gradient of the mobile phase. The effluent was monitored at 220 nm and subsequently by analytical HPLC. Relevant fractions were pooled, evaporated and lyophilized. The identity of the synthetic peptides was verified by analytical reverse phase chromatography and by amino acid analysis. A solution of potassium ferricyanide (0.01M, pH 7.0) was added slowly to a dilute aqueous solution (0.5 mM) of the linear peptide at pH 7.0. After 24 hours at room temperature, the pH was lowered to 5.0 and the solution treated with ion exchange resin (Bio-Rad Ag-3-X4a, Cl-form) for 30 minutes. The suspension was filtered and the filtrate lyophilized to give the crude cyclic peptide. The peptide was purified by preparative reverse phase HPLC and characterized by amino acid analysis. Proof of cyclicity was obtained by comparing the HPLC mobility of the cyclic peptide with the starting linear peptide by reducing an aliquot of the cyclic peptide back to the linear peptide and also by observing the disappearance of free sulfhydryl groups (Ellman's Test) after the cyclization. Peptides were conjugated to BSA or KLH previously derivatized with either sulfosuccinimidyl 4-(p-maleimidophenyl)butyrate (Sulfo-SMPB) or sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (Sulfo-SMCC). An aqueous solution of sulfo-SMPB or sulfo-SMCC (Pierce Chemicals) was added to a solution of BSA or KLH in 0.02 M sodium phosphate buffer (pH 7.0). The mixture was shaken at room temperature for 45 minutes and the activated carrier immediately applied to a Sephadex G-25 column equilibrated with 0.1M sodium phosphate buffer (pH 6.0) at 4� C. The fractions of the first peak absorbance (280 nm) corresponding to activated carrier were combined in a round bottom flask to which was added a solution of peptide in 0.05 M sodium phosphate buffer (pH 6.2). The mixture was thoroughly flushed with N2 and incubated overnight at room temperature. The coupling efficiency was monitored using 3H-labeled peptide and by amino acid analysis of the conjugate. Each well of the microtiter plate is saturated with 100 μl of a solution containing a peptide or mixture of peptides (5 μg/ml) and left overnight. The wells are emptied and washed twice with a washing buffer (Tris, 0.043M; NaCl, 0.5M; thimerosal, 0.01% w/v; Tween 20, 0.05% v/v; pH 7.4). The wells are then saturated with 0.35 ml of washing buffer for 1 hr. at 37� C. and washed once with the same buffer. Serum samples to be analyzed are diluted with specimen buffer (washing buffer plus casein, 0.05% w/v). The wells are rinsed with washing buffer prior to the addition of the diluted serum sample (0.1 ml). These are left to incubate for 1 hr. at room temperature. The wells are then emptied, washed twice rapidly and then once for two minutes with washing buffer. The conjugate solution (affinity purified goat antibody to human IgG peroxidase labeled, 0.5 mg in 5 ml 50% glycerol) diluted with 1% w/v bovine serum albumin in washing buffer is added to each well (0.1 ml) and incubated for 1 hr. at room temperature. The wells are then emptied and washed twice rapidly with washing buffer and then five times in which the buffer was in contact with the well 2 minutes per washing. The substrate solution (3,3′,5,5′-tetramethylbenzidine, 8 mg per ml of DMSO) is diluted with 100 volumes 0.1M citrate-acetate buffer, pH 5.6 containing 0.1% v/v of 30% H2O2 and added to each well (0.1 ml per well). After 10 minutes the contents of each well is treated with 0.1 ml 2N H2SO4 and the optical density read at 450 nm. All determinations are done in duplicate. Peptide cocktails were prepared by mixing together equal volumes of two peptide solutions each at 10 ug/ml. One cocktail used peptides BCH-87c and BCH-202c and the other cocktail was a mixture of BCH-87ck and BCH-202ck. Each cocktail was used to coat two series of plates as described earlier. Patent Citations
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Poll results from last week's question regarding anemia management in the dialysis patient. Interestingly, there was a fairly strong consensus on this one, with over 75% of individuals electing to give a course of iv iron to the dialysis patient in question, who was anemic despite relatively large EPO doses and a ferritin level of 900. Be sure and check out the newest poll question in the right margin!
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My go to Magic store for all I need! New amazments added all the time, endless selection & points earned for discounts. Also want to say, super fast...Read More ->
Great Prices, Great Service! I ordered here twice and I got both of what I needed by 2 DAYS! I'll keep buying here.Read More ->


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Imagine this happening in Washington: Alex Ovechkin shows up at Verizon Center coughing and wheezing due to bronchitis and a high fever.
Its one thing for Matt Bradley to go on a Toronto radio station and say what everyone else had been saying about Alexander Semin for years that his questionable work ethic prevented him from being
Former Washington Capitals defenseman Scott Hannan has agreed to one-year 1 million deal with the Nashville Predators.
Alexander Semin finished his career in Washington with 197 goals, fifth on the clubs all-time list behind Peter Bondra 472, Mike Gartner 397, Alex Ovechkin 339 and Mike Ridley 218.
Saturday, February 13, 2010
By Joe Beninati
Capitals Play-by-Play


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And that's that for another memorable Masters. Hope you enjoyed the coverage, be sure to follow the live commentary from our man at the US Open in June. Congratulations Bubba.
00:38 -
BUBBA WATSON, MASTERS CHAMPION! Two cautious putts, and cue the waterworks. Just a week after he and his wife adopted their first child, Watson is now celebrating his first major title. Emotional scenes as Bubba is embraced by the likes of Ben Crane, Aaron Baddeley, Stewart Cink and Rickie Fowler.
00:35 -
Fantastic effort from Oosthuizen, but his par putt just runs out of pace and trickles across the face of the hole. Very unlucky, and it looks like Bubba is home and hosed.
00:33 -
Oosthuizen chips from the front edge, but it's a touch heavy and clings onto the back fringe, a full 15 feet past the hole. He has to find one more putt, or else Bubba has two putts from nowhere.
00:30 -
AWESOME! Watson breaks his wrists with perfect timing to draw his ball at least 40 yards around the trees to find the green, and he has around 10 feet left for birdie. A shot of truly stunning quality.
00:28 -
Oosthuizen makes a good connection from the first cut and it looks great in the air, but it ha't quite got the legs and pitches in the steep bank at the front of the green. Good result, but Watson does have a shot, although he needs to give it a mammoth draw.
00:26 -
The plot thickens. Bubba goes for the big fade but double-crosses himself and pulls his drive deep into the pines. Oosthuizen plucks out a three-wood, but he carves his tee shot to the right as well. He's not in the timber, but he's got an awfully long second.
00:22 -
OH BUBBA! No-one has seen Watson smile all day, but he can't resist a wry grin as his putt for the Masters does the opposite to Oosthuizen's and misses to the left. More drama to come as they head for the 10th.
00:20 -
SO CLOSE! Oosthuizen's putt looks bang on line, but it veers right over the last six feet and can't catch enough lip.
00:20 -
But what a reply from Bubba, who hits a smooth wedge from 131 yards over the pin, and it has enough backspin to get to the slope and he has around 10 feet for a three. Advantage Watson.
00:20 -
Oosty's second leaks a little right, but it hits the right contours and arches round towards the hole, settling 15 feet away. Great shot.
00:15 -
Oosthuizen backs off, has a quick towel down, steps up again and gives it the full treatment. It flirts with Sandy's bunker, but it clings to the short grass and he's not far behind Bubba's.
00:10 -
Here we go then. Bubba has the honour and hits another high fade over the trees to within a couple of yards of the divot he created 10 minutes ago.
00:10 -
It's been yet another final day of history and high drama at Augusta, and it's not over yet. Oosthuizen and Watson will head back to the 18th tee in a moment, and the 10th is next should they halve the last.
00:05 -
Hanson breaks into a wry smile after he belatedly gets a mid-range putt to drop. That's a closing birdie, but a 73 is two shots too many as Oosthuizen and Watson prepare for a sudden-death play-off.
00:02 -
A two-putt par to finish for Mickelson, and a disappointing 72 leaves him two off the pace. He had better have a good explanation for his adventures at the fourth!
00:02 -
Mickelson has time for one more magical moment, a massive draw from the pines to within 20 feet at the last. Hanson knocks his a couple of yards closer.
00:00 -
Don't leave it short, don't leave it short ...... it's short. Mickelson knows he needs birdie-birdie to extend his tournament, but his 25-footer rolls 24 feet at the 17th.
23:55 -
What pressure? Oosthuizen has been sensational with the flat-stick today, definite shades of Retief Goosen on the final day of the US Open at Shinnecock in 2004. Oosty holds his nerve and drops the putt in the left edge, and a play-off is forthcoming.
23:55 -
Oosthuizen's birdie effort is on line for most of the trip, but it veers left and trundles four feet past. Watson's stays on the high side, but it's a tap-in for the safest of pars.
Show All


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Journal of Hospital Administration (ISSN 1927-6990 EISSN 1927-7008) is an international, open access, and peer-reviewed scientific journal published by Sciedu Press. It is devoted to publishing research papers in the fields of managing practice and research in all branches of hospital administration. JHA welcomes original articles and reviews. It is published in both online and printed versions.
JHA is included in:
- EBSCOhost
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Areas include but are not limited to:
- Healthcare quality and patient safety
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To facilitate rapid publication and minimize administrative costs, JHA accepts Online submission and Email submission. All manuscripts and any supplementary material can be submitted via the journal's Online Submission and peer-review system or email to email@example.com. For online submission, please create a new account and then follow the instructions given.
Original Articles, Reviews.
2014: Articles Received: 111; Accepted: 65; Rejected: 34; Published: 72; Retracted: 0
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2013 Q4: Articles Received: 27; Accepted: 17; Rejected: 3; Published: 20; Retracted: 0
2013 Q3: Articles Received: 32; Accepted: 17; Rejected: 7; Published: 13; Retracted: 0
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e-Version First is a feature offered through our journal platform. It allows PDF version of manuscripts that have been peer reviewed and accepted, to be hosted online prior to their inclusion in a final printed journal. Readers can freely access or cite the article. We aim to publish accepted manuscripts in e-Version First in two week's time after the final draft completed.
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The editors of JHA would like to thank all of the reviewers who made excellent work and great contribution to our journal in 2014. Without the reviewers' professional kowledge, skill and great help, JHA could not achieve such high standards. Please accept our heartfelt gratitude and wishes. May joy, peace, and happiness be with you and your entire family always! | |
Posted: 2015-01-01 | |
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Vol 4, No 2 (2015)
Journal of Hospital Administration, Vol. 4, No. 2, April 2015, e-Version First TM
Table of Contents
Original Article
The association between the transfer of emergency department admitted patients to inpatient hallways and outcomes of oncology patients | |
Charles Lim, Matthew C. Cheung, Maureen E. Trudeau, Kevin R. Imrie, Ben de Mendonca, Simron Singh | p1 |
Clinical learning environment in government and private hospitals in relation to gender in Karachi, Pakistan - a comparative study | |
Sharmeen Amin, Zafeer Ahmed Yazdani, Ghazala Usman | p9 |
Predicting hospital length of stay for geriatric and adult patients with schizophrenia | |
Zahinoor Ismail, Tamara Arenovich, Charlotte Grieve, Peggie Willett, Donald Addington, Tarek K Rajji, Benoit H. Mulsant | p15 |
Preliminary development of a reference conceptual framework about social responsibility of public hospitals in China | |
Lizheng Shi, Fei Liang, Hui Shao, Xianzhi Hu, Qian Gu, Wenbin Liu, Xiong Ke, Thomas Jefferson Stranova, Yingyao Chen | p23 |
Are we teaching our hospitalized patients self-management skills? Opportunities lost | |
Serena Miotto, Lynn McNicoll, Kristen Butterfield, Christina Rincon, Mriganka Singh, Stefan Gravenstein | p30 |
Bundled payments for care improvement initiative – insights from the test pilots of payment reform | |
Jason M Sutherland, William B. Borden | p37 |
Smoke free site and service awareness amongst hospital staff: a survey in an acute cancer centre | |
Peter Mackereth, Maycock Paula, Orrett Linda | p43 |
Team strategies and tools to enhance performance and patient safety training: The effect of training on both nursing staff perceptions regarding physician behaviors and patient satisfaction scores in the ED | |
Shari I. Brand, Kristen M. Slee, Yu-Hui Chang, Meng-Ru Cheng, Christopher A. Lipinski, Rick R. Arnold, Stephen J.Traub | p48 |
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EDIT: Thanks to David Ruttka, I was able to figure it out after looking at LabelExtensions.cs from Mvc3's RTM release.
For the field name: string field = ExpressionHelper.GetExpressionText(expression);
For the model, I needed to specify the model I wanted to cast as for the Helper- where TModel:Foo then I could get at the model: BarTypeEnum barType = ((Foo)html.ViewData.Model).BarType;
I've updated the source below to what's working for me.
I'm attempting to create a html helper function similar to LabelFor in Mvc3 to return a string value based on Foo.BarType and the name of Foo's field passed in from the html.
In the function FooLabelFor below, how do I get model and field name passed into the function?
I went looking for the source for System.Web.Mvc.HtmlLabelFor, but was unable to find it in the Mvc3 source code.
//model class
public class Foo
public string Bar { get; set; }
public BarTypeEnum BarType { get; set; }
//html helper class
public static class HtmlHelpers {
public static string FooLabelFor<TModel, TValue>(this HtmlHelper<TModel> html, Expression<Func<TModel, TValue>> expression) where TModel:Foo
BarTypeEnum barType = ExpressionHelper.GetExpressionText(expression);
string field = ((Foo)html.ViewData.Model).BarType;
return GlobalizeText(enumHelper.stringvalue(barType), field);
@model Foo
<div>@Html.FooLabelFor(m => m.Bar)</div>


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Dr. Austin named NAE member
Dr. Wanda M. Austin, Advisory Board Member
Dr. Wanda M. Austin, a member of our department's Advisory Board, and also President and CEO of the Aerospace Corporation, has been named as a National Academy of Engineering member for her contributions and leadership in the national intelligence community.
Austin has received numerous awards and citations. Among them are the Air Force Scroll of Achievement, the National Reconnaissance Office Gold Medal, the U.S. Air Force Meritorious Civilian Service Medal, the Air Force Space and Missile Systems Center's Martin Luther King Spirit of the Dream Award, the Society of Women Engineers Upward Mobility Award, and the Women in Aerospace Outstanding Achievement Award.
Austin earned a Bachelor of Arts degree in mathematics from Franklin and Marshall College, Masters of Science degrees in both mathematics and systems engineering from the University of Pittsburgh, and a PhD in systems engineering from the University of Southern California. She is a member of the NASA Advisory Council and is a fellow of the AIAA (American Institute of Aeronautics and Astronautics).
More information available at the NAE web site:
Related Articles: Aerospace Students Excel at 14th AIAA YPSE ConferenceLauren Trollinger Awarded a WIA Foundation—AIAA Digital Avionics ScholarshipFive Aerospace Engineering Faculty Promoted to AIAA Associate FellowsAerospace Engineering Students Win at the AIAA Student Paper ConferenceFive Aerospace Engineering Students Win Honors at AIAA ConferenceUndergraduate Students Sweep AIAA YPSE ConferenceAerospace Students Shine at the 11th AIAA YPSE ConferenceAerospace Students Earn Accolades at AIAA Mid-Atlantic Student ConferenceGeerts Earns First Place at 2011 AIAA Regional Symposium in PhiladelphiaProfessor Wereley honored with AIAA Sustained Service Award February 9, 2008
Current Headlines
Fourney Named ASME Fellow
Lagor and Sherman Named 2014 Future Faculty Inductees
Stacy Sidle Awarded Aerospace Corporation Fellowship
Hartzell Receives Astronomical Honor
Flatau Promoted to AIAA Fellow
Clark School Welcomes Kier, New Associate Director of Development
Paley Awarded 2014 E. Robert Kent Teaching Award for Junior Faculty
UMD Clark School Highlights Disaster Resilience at Mpact Week
Vera Klimchenko Wins ASME Student Paper Award
University of Maryland Breaks Ground for A. James Clark Hall


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Rajon Rondo has been a suspended one game for "conduct detrimental to the team" after he got into a squabble with Mavericks coach Rick Carlisle last night. He'll miss tonight's game against the Hawks.
Rick Carlisle Gets The Business From Rajon Rondo, Reporter
Dallas Mavericks head coach Rick Carlisle did not have the best day at the office yesterday. His team came away with a 99-92 victory over the Toronto Raptors, which was good, but he also got into a shouting match with starting point guard Rajon Rondo in the middle of the game, which was very bad. Then, he just about…
Steph Curry Has A Steph Curry Game
If you thought anything at all like "this Warriors-Mavericks game tonight isn't worth staying up late for," you should immediately smack yourself in the head. Hard. It has been true for at least a season now, but the point was driven home especially forcefully tonight: The Golden State Warriors are always worth your…
Former Mavs Big Man Roy Tarpley Passes Away At 50
Roy Tarpley, the talented Dallas Mavericks power forward who flamed out of the NBA due to substance abuse issues, died Friday, at the age of 50. According to The Dallas Morning News there was no official cause of death, but multiple Mavericks officials were informed he had liver failure.
That's One Way To Get Business Done
From #Colbert Farewell: @MarkCuban finalized Rondo deal in green room, sitting w/Waterston, Daniels, me + dismaying UN Amb. Samantha Power
— Keith Olbermann (@KeithOlbermann)
December 19, 2014
That Rondo-To-Dallas Trade Is Going Down
What's half a season of (and exclusive negotiating rights to) Rajon Rondo worth? The Celtics and Mavericks appear to be in the final stages of figuring that out. Marc Stein has the purported haul:
This Looks Like The Year The Celtics Finally Trade Rajon Rondo
Swapping Rajon Rondo trade rumors is America's oldest and most treasured holiday tradition. But this year, the Celtics' last chance to get something for their star point guard, it seems like it's actually going to happen. Last night, all around the same time, seemingly every NBA beat guy began reporting that talks on…
Monta Ellis's Game-Winning Buzzer-Beater Was Bananas
I think it's time to admit that there probably isn't another player in the NBA who is better at hitting reckless, off-balance buzzer-beaters than Monta Ellis. Almost two years after hitting a game-winner so absurd that Ellis himself had to flee from it, the Mavericks shooting guard once again did his thing against the…
Kirk Hinrich Submits Contender For Dumbest Foul Of The Season
Two weeks ago I wrote that Alex Len had committed the dumbest foul of the season. Tonight Kirk Hinrich announced his challenge to the throne.
Anthony Davis Is A Goddamn Demon
Anthony Davis is going to own the league sooner rather than later, because Anthony Davis is a shot-devouring, rebound-snatching, dunking monster who is now armed with a solid mid-range jumper and respectable handles. Dunks and smooth jumpers and blocks are great, but if you really want to see what makes Anthony Davis…
Chandler Parsons Is Probably Thrilled To Be Away From Dwight Howard
First, watch this video of Howard catching up with his old teammate before last night's preseason game.
Mark Cuban Suggests An NBA World Cup
Dallas Mavericks owner and naked billionaire Mark Cuban has long been against the NBA's partnership with FIBA and the International Olympic committee, and Paul George's injury in Friday night's Team USA scrimmage gave him another opportunity to rip the current system of international play.
A Good Offer Sheet Leaves Everyone Pissed Off
Of the three interested parties, no one really expected Chandler Parsons to be a Dallas Maverick, and I'm not convinced anyone, even the Mavs, wanted it to happen. But after everything clicked into place in the convoluted clockwork of NBA free agency, the Rockets declined to shell out $15 million a year for the…


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With economic analysts sending mixed messages concerning the 2003 construction season, producers aren't quite sure what the immediate future will bring. Even so, it's a sure bet that many of the industry innovators will include a visit to the World of Concrete as a way to hedge their losses. That's because it's the place where they can learn not only about innovative production processes but gain insights into what their customers need.
Industry Insights Schedule
Tuesday, February 4, 2:00–3:00 p.m.
Wednesday, February 5, 2:00–3:00 p.m.
The National Ready Mixed Concrete Association has released a newly revised Pervious...
This is the first time in the five-year history of the award program that dual winners...
Portland Cement Association
World of Concrete
Education and Seminars
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Publications (26)100.91 Total impact
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ABSTRACT: Small ubiquitin-like modifier (SUMO) post-translational modification (PTM) of proteins has a crucial role in the regulation of important cellular processes. This protocol describes the chemical synthesis of functional SUMO-peptide conjugates. The two crucial stages of this protocol are the solid-phase synthesis of peptide segments derivatized by thioester or bis(2-sulfanylethyl)amido (SEA) latent thioester functionalities and the one-pot assembly of the SUMO-peptide conjugate by a sequential native chemical ligation (NCL)/SEA native peptide ligation reaction sequence. This protocol also enables the isolation of a SUMO SEA latent thioester, which can be attached to a target peptide or protein in a subsequent step. It is compatible with 9-fluorenylmethoxycarbonyl (Fmoc) chemistry, and it gives access to homogeneous, reversible and functional SUMO conjugates that are not easily produced using living systems. The synthesis of SUMO-peptide conjugates on a milligram scale takes 20 working days.Nature Protocols 02/2015; 10(2):269-92. · 7.78 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: The regulation of PfPP1 phosphatase activity in Plasmodium falciparum remains to be deciphered. Data from homologous eukaryotic PP1 phosphatases suggest that several protein regulators should be involved in this essential process. One such regulator, named PfI2 based on its primary sequence homology with eukaryotic I2, was recently shown to be able to interact with PfPP1 and to inhibit its phosphatase activity, mainly through the canonical "RVxF" binding motif. The details of the structural and functional characteristics of this interaction are investigated here. Using NMR spectroscopy, a second site of interaction is suggested to reside between residues D94-T117 and contains the "FxxR/KxR/K" binding motif present in other I2 proteins. This site seems to play in concert/synergy with the "RVxF" motif to bind PP1, as only mutations in both motifs were able to abolish completely this interaction. However, regarding the structure/function relationship, the mutation of either "RVxF" or "FxxR/KxR/K" motif is more drastic, as each mutation prevents the capacity of PfI2 to trigger GVBD in micro-injected Xenopus oocytes. This indicates that the tight association of the PfI2 regulator to the PP1 phosphatase, mediated by a two-site interaction, is necessary to exert its function. Based on these results, the use of a peptide derived from the "FxxR/KxR/K" PfI2 motif was investigated for its potential effect on Plasmodium growth. This peptide, fused at its N-terminus to a penetrating sequence, was shown to accumulate specifically in infected erythrocytes and to have an anti-plasmodial effect.This article is protected by copyright. All rights reserved.Structured digital abstractPfPP1 and PfI2 bind by nuclear magnetic resonance (1, 2)PP1 physically interacts with PfPP1 by anti tag coimmunoprecipitation (View interaction)PfI2 binds to PfPP1 by enzyme linked immunosorbent assay (View interaction)PfI2 binds to PfPP1 by surface plasmon resonance (1, 2, 3, 4)FEBS Journal 08/2014; · 3.99 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: The capacity of many proteins to interact with natural or synthetic polyanions has been exploited for modulating their biological action. However, the polydispersity of these macromolecular polyanions as well as their poor specificity is a severe limitation to their use as drugs. An emerging trend in this field is the synthesis of homogeneous and well-defined polyanion-peptide conjugates which act as bivalent ligands with the peptide part bringing the selectivity of the scaffold. Alternately, this strategy can be used for improving the binding of short peptides to polyanion-binding protein targets. This work describes the design and first synthesis of homogeneous polysulfonate-peptide conjugates using thiocarbamate ligation for binding to the extracellular domain of MET tyrosine kinase receptor for hepatocyte growth factor.Bioconjugate Chemistry 04/2014; · 4.82 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: A bis(2-sulfanylethyl)amino PEG-based resin enabled the synthesis of large (50 Aa) SEA or thioester peptides using Fmoc-SPPS. These peptide segments permitted the first total synthesis of a 97 amino-acid long SUMO-1-SEA peptide thioester surrogate and of a functional and reversible SUMO-1 peptide conjugate.Chemical Science 01/2014; 5(5):2017. · 8.60 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: BACKGROUND: Receptor tyrosine kinases (RTK) form a family of transmembrane proteins widely conserved in Metazoa, with key functions in cell-to-cell communication and control of multiple cellular processes. A new family of RTK named Venus Kinase Receptor (VKR) has been described in invertebrates. The VKR receptor possesses a Venus Fly Trap (VFT) extracellular module, a bilobate structure that binds small ligands to induce receptor kinase activity. VKR was shown to be highly expressed in the larval stages and gonads of several invertebrates, suggesting that it could have functions in development and/or reproduction. RESULTS: Analysis of recent genomic data has allowed us to extend the presence of VKR to five bilaterian phyla (Platyhelminthes, Arthropoda, Annelida, Mollusca, Echinodermata) as well as to the Cnidaria phylum. The presence of NveVKR in the early-branching metazoan Nematostella vectensis suggested that VKR arose before the bilaterian radiation. Phylogenetic and gene structure analyses showed that the 40 receptors identified in 36 animal species grouped monophyletically, and likely evolved from a common ancestor. Multiple alignments of tyrosine kinase (TK) and VFT domains indicated their important level of conservation in all VKRs identified up to date. We showed that VKRs had inducible activity upon binding of extracellular amino-acids and molecular modeling of the VFT domain confirmed the structure of the conserved amino-acid binding site. CONCLUSIONS: This study highlights the presence of VKR in a large number of invertebrates, including primitive metazoans like cnidarians, but also its absence from nematodes and chordates. This little-known RTK family deserves to be further explored in order to determine its evolutionary origin, its possible interest for the emergence and specialization of Metazoa, and to understand its function in invertebrate development and/or reproductive biology.BMC Genomics 05/2013; 14(1):361. · 4.04 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: Hepatocyte growth factor/scatter factor (HGF/SF) is the high affinity ligand of MET tyrosine kinase receptor. We report here the total synthesis of a biotinylated analogue of human HGF/SF N domain. Functionally, N domain is part of the HGF/SF high affinity binding site for MET and also the main HGF/SF binding site for heparin. The 97 Aa linear chain featuring a C-terminal biotin group was assembled in high yield using an N-to-C one-pot three segments assembly strategy relying on a sequential Native Chemical Ligation (NCL)/bis(2-sulfanylethyl)amido (SEA) native peptide ligation process. The folded protein displayed the native disulfide bond pattern and showed the ability to bind heparin.Bioorganic & medicinal chemistry 03/2013; · 2.82 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: The GRB2-associated binder 1 (GAB1) docking/scaffold protein is a key mediator of the MET-tyrosine kinase receptor activated by hepatocyte growth factor/scatter factor (HGF/SF). Activated MET promotes recruitment and tyrosine phosphorylation of GAB1, which in turn recruits multiple proteins and mediates MET signaling leading to cell survival, motility, and morphogenesis. We previously reported that, without its ligand, MET is a functional caspase target during apoptosis, allowing the generation of a p40-MET fragment that amplifies apoptosis. In this study we established that GAB1 is also a functional caspase target by evidencing a caspase-cleaved p35-GAB1 fragment that contains the MET binding domain. GAB1 is cleaved by caspases before MET, and the resulting p35-GAB1 fragment is phosphorylated by MET upon HGF/SF binding and can interact with a subset of GAB1 partners, PI3K, and GRB2 but not with SHP2. This p35-GAB1 fragment favors cell survival by maintaining HGF/SF-induced MET activation of AKT and by hindering p40-MET pro-apoptotic function. These data demonstrate an anti-apoptotic role of caspase-cleaved GAB1 in HGF/SF-MET signaling.Journal of Biological Chemistry 08/2012; 287(42):35382-96. · 4.60 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: The receptor tyrosine kinase Met and its ligand, the hepatocyte growth factor/scatter factor, are essential for embryonic development, whereas deregulation of Met signaling pathways is associated with tumorigenesis and metastasis. The presenilin-regulated intramembrane proteolysis (PS-RIP) is involved in ligand-independent downregulation of Met. This proteolytic process involves shedding of the Met extracellular domain followed by γ-secretase cleavage, generating labile intracellular fragments degraded by the proteasome. We demonstrate here that upon shedding both generated Met N- and C-terminal fragments are degraded directly in the lysosome, with C-terminal fragments escaping γ-secretase cleavage. PS-RIP and lysosomal degradation are complementary, because their simultaneous inhibition induces synergistic accumulation of fragments. Met N-terminal fragments associate with the high-affinity domain of HGF/SF, confirming its decoy activity which could be reduced through their routing to the lysosome at the expense of extracellular release. Finally, the DN30 monoclonal antibody inducing Met shedding promotes receptor degradation through induction of both PS-RIP and the lysosomal pathway. Thus, we demonstrate that Met shedding initiates a novel lysosomal degradation which participates to ligand-independent downregulation of the receptor.Traffic 06/2012; 13(9):1261-72. · 4.71 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: Growing evidence indicates that the protein regulators governing protein phosphatase 1 (PP1) activity have crucial functions because their deletion drastically affects cell growth and division. PP1 has been found to be essential in Plasmodium falciparum, but little is known about its regulators. In this study, we have identified a homolog of Inhibitor-3 of PP1, named PfI3. NMR analysis shows that PfI3 belongs to the disordered protein family. High affinity interaction of PfI3 and PfPP1 is demonstrated in vitro using several methods, with an apparent dissociation constant KD of 100 nm. We further show that the conserved 41KVVRW45 motif is crucial for this interaction as the replacement of the Trp45 by an Ala45 severely decreases the binding to PfPP1. Surprisingly, PfI3 was unable to rescue a yeast strain deficient in I3 (Ypi1). This lack of functional orthology was supported as functional assays in vitro have revealed that PfI3, unlike yeast I3 and human I3, increases PfPP1 activity. Reverse genetic approaches suggest an essential role of PfI3 in the growth and/or survival of blood stage parasites because attempts to obtain knock-out parasites were unsuccessful, although the locus of PfI3 is accessible. The main localization of a GFP-tagged PfI3 in the nucleus of all blood stage parasites is compatible with a regulatory role of PfI3 on the activity of nuclear PfPP1.Journal of Biological Chemistry 01/2012; 287(2):1306-1321. · 4.60 Impact Factor -
Article: Polysaccharide microarrays: application to the identification of heparan sulphate mimetics.
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ABSTRACT: The interaction of polysaccharides with proteins modulates or triggers many biological effects. In particular, heparan sulphate proteoglycans (HSPGs) have multiple regulatory interactions with growth factors, enzymes, enzyme inhibitors, and some components of the extracellular matrix. The important role played by HSPGs has motivated the synthesis and selection of HSPG mimetics for modulating the biological activity of HS-binding proteins. We present hereinafter an efficient polysaccharide microarray method that allows the screening of HS-mimetic libraries towards HS-binding growth factors, a major class of polypeptides whose inhibition or potentiation is of high medical interest.Methods in molecular biology (Clifton, N.J.) 01/2012; 808:231-40. · 1.29 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: Growing evidence indicates that the protein regulators governing protein phosphatase 1 (PP1) activity have crucial functions because their deletion drastically affects cell growth and division. PP1 has been found to be essential in Plasmodium falciparum, but little is known about its regulators. In this study, we have identified a homolog of Inhibitor-3 of PP1, named PfI3. NMR analysis shows that PfI3 belongs to the disordered protein family. High affinity interaction of PfI3 and PfPP1 is demonstrated in vitro using several methods, with an apparent dissociation constant K(D) of 100 nm. We further show that the conserved (41)KVVRW(45) motif is crucial for this interaction as the replacement of the Trp(45) by an Ala(45) severely decreases the binding to PfPP1. Surprisingly, PfI3 was unable to rescue a yeast strain deficient in I3 (Ypi1). This lack of functional orthology was supported as functional assays in vitro have revealed that PfI3, unlike yeast I3 and human I3, increases PfPP1 activity. Reverse genetic approaches suggest an essential role of PfI3 in the growth and/or survival of blood stage parasites because attempts to obtain knock-out parasites were unsuccessful, although the locus of PfI3 is accessible. The main localization of a GFP-tagged PfI3 in the nucleus of all blood stage parasites is compatible with a regulatory role of PfI3 on the activity of nuclear PfPP1.Journal of Biological Chemistry 11/2011; 287(2):1306-21. · 4.60 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: Three in one: Native chemical ligation (NCL) and bis(2-sulfanylethyl)amido (SEA) ligation allow the one-pot assembly of three peptide segments in the N-to-C direction. The SEA group is switched off by intramolecular disulfide bond formation during NCL. Then, a phosphine switches it on to trigger the second SEA ligation step. The K1 domain of the hepatocyte growth factor was synthesized and found to be biologically active.Angewandte Chemie International Edition 11/2011; 51(1):209-13. · 11.34 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: Schistosomiasis (bilharzia) is a parasitic disease of worldwide significance affecting human and animals. As schistosome eggs are responsible for pathogenesis, the understanding of processes controlling gonad development might open new perspectives for intervention. The Src-like tyrosine-kinase SmTK3 of Schistosoma mansoni is expressed in the gonads, and its pharmacological inhibition reduces mitogenic activity and egg production in paired females in vitro. Since Src kinases are important signal transduction proteins it is of interest to unravel the signaling cascades SmTK3 is involved in to understand its cellular role in the gonads. Towards this end we established and screened a yeast two-hybrid (Y2H) cDNA library of adult S. mansoni with a bait construct encoding the SH3 (src homology) domain and unique site of SmTK3. Among the binding partners found was a diaphanous homolog (SmDia), which was characterized further. SmDia is a single-copy gene transcribed throughout development with a bias towards male transcription. Its deduced amino acid sequence reveals all diaphanous-characteristic functional domains. Binding studies with truncated SmDia clones identified SmTK3 interaction sites demonstrating that maximal binding efficiency depends on the N-terminal part of the FH1 (formin homology) domain and the inter-domain region of SmDia located upstream of FH1 in combination with the unique site and the SH3 domain of SmTK3, respectively. SmDia also directly interacted with the GTPase SmRho1 of S. mansoni. In situ hybridization experiments finally demonstrated that SmDia, SmRho1, and SmTK3 are transcribed in the gonads of both genders. These data provide first evidence for the existence of two cooperating pathways involving Rho and Src that bridge at SmDia probably organizing cytoskeletal events in the reproductive organs of a parasite, and beyond that in gonads of eukaryotes. Furthermore, the FH1 and inter domain region of SmDia have been discovered as binding sites for the SH3 and unique site domains of SmTK3, respectively.PLoS ONE 09/2009; 4(9):e6998. · 3.53 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: Very little is known about insulin signalling in schistosomes despite its potential importance in host-parasite molecular dialogue and parasite growth and development. The recent characterization of two insulin receptors (SmIR-1 and SmIR-2) in Schistosoma mansoni has led us to reconsider the question of the potential importance of insulin in host-schistosome interactions. In this work, we demonstrated that insulin could regulate glucose uptake in schistosomes and we investigated the implication of SmIR-1 and SmIR-2 in this process. The possibility that specific inhibitors of SmIR-1 and SmIR-2 tyrosine kinase activities could be developed to target schistosomes is discussed.Parasite 01/2009; 15(4):573-9. · 0.82 Impact Factor -
Article: Diversification of the insulin receptor family in the helminth parasite Schistosoma mansoni.
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ABSTRACT: Insulin signalling is a very ancient and well conserved pathway in metazoan cells, dependent on insulin receptors (IR) which are transmembrane proteins with tyrosine kinase activity. A unique IR is usually present in invertebrates whereas two IR members are found with different functions in vertebrates. This work demonstrates the existence of two distinct IR homologs (SmIR-1 and SmIR-2) in the parasite trematode Schistosoma mansoni. These two receptors display differences in several structural motifs essential for signalling and are differentially expressed in parasite tissues, suggesting that they could have distinct functions. The gene organization of SmIR-1 and SmIR-2 is similar to that of the human IR and to that of the IR homolog from Echinococcus multilocularis (EmIR), another parasitic platyhelminth. SmIR-1 and SmIR-2 were shown to interact with human pro-insulin but not with pro-insulin-like growth factor-1 in two-hybrid assays. Phylogenetic results indicated that SmIR-2 and EmIR might be functional orthologs whereas SmIR-1 would have emerged to fulfil specific functions in schistosomes.FEBS Journal 03/2007; 274(3):659-76. · 3.99 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: In order to characterize protein cofactors of the Schistosoma mansoni nuclear receptor SmFtz-F1, we have screened a yeast two-hybrid adult worm cDNA library using a construct expressing the D, E and F domains of SmFtz-F1 as bait. One of the selected clones encoded a sequence without homologues in any other species, apart from Schistosoma japonicum. The complete sequence was obtained by 5' and 3' rapid amplification of cDNA ends (RACE) and comprised 3660 nucleotides with an open reading frame of 788 amino acids. The gene is expressed at all schistosome life cycle stages at a 5-11-fold higher level than SmFtz-f1. The protein, named SmFIP-1, interacted with SmFtz-F1 in a GST pull-down assay and in a mammalian two-hybrid assay in CV-1 cells. Although SmFIP-1 contains a consensus NR box (LXXLL) this was not involved in the interaction with SmFtz-F1. However, interaction did depend on the AF2-AD motif in the nuclear receptor ligand binding domain. Deletion analysis showed that the C-terminal moiety of SmFIP-1 was involved in the binding, but this could not be localized to a particular motif, suggesting that the binding may be conformation-dependent. Finally, SmFIP-1 markedly repressed SmFtz-F1-mediated transcription in a dose-dependent manner from the SmFtz-f1 gene promoter demonstrating that SmFIP-1 is a schistosome-specific transcriptional corepressor.Molecular and Biochemical Parasitology 08/2006; 148(1):10-23. · 2.24 Impact Factor -
Article: Glyceroneogenesis: an unexpected metabolic pathway for glutamine in Schistosoma mansoni sporocysts.
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ABSTRACT: To date, glyceroneogenesis has only been described in mammals but we demonstrate in this paper that it could exist in the invertebrate Schistosoma mansoni, the parasitic helminth transmitted by fresh water molluscs and responsible for the major human endemic disease, schistosomiasis. Glyceroneogenesis is a phosphoenolpyruvate carboxykinase (PEPCK)-dependent process by which glycerol can be produced from precursors like glutamine and therefore represents a truncated gluconeogenic pathway. We have previously demonstrated the possible central role of glutamine in mollusc-schistosome interactions. In this work, we show that glutamine effectively promotes in vitro survival and protein synthesis in sporocysts, the intramolluscan larval stages of S. mansoni, possibly through its role as an energy source. However, the demonstration that PEPCK is massively expressed in these larval forms as compared to adult parasites, together with the observation that 3-mercaptopicolinate, a specific inhibitor of PEPCK, significantly reduces the effect of glutamine on sporocyst growth, suggest that glutamine could also be used for glucose or glycerol production. Results of [14C] glutamine incorporation confirmed that neosynthesis of glucose and mainly of glycerol occurred in sporocysts and was dependent on PEPCK activity. Therefore, our results strongly indicate that glyceroneogenesis could exist in schistosomes. Several hypotheses can be proposed concerning the importance of glycerol for the adaptation of this helminth to its host osmotic and energetic environment.Molecular and Biochemical Parasitology 07/2006; 147(2):145-53. · 2.24 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: Parasitic helminths remain major pathogens of both humans and animals throughout the world. The success of helminth infections depends on the capacity of the parasite to counteract host immune responses but also to exploit host-derived signal molecules for its development. Recent progress has been made in the characterization of growth factor receptors of various nematode and flatworm parasites with the demonstration that transforming growth factor beta (TGF-beta), epidermal growth factor (EGF) and insulin receptor signalling pathways are conserved in helminth parasites and potentially implicated in the host-parasite molecular dialogue and parasite development.FEBS Letters 06/2006; 580(12):2968-75. · 3.34 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: Interleukin (IL-)7 and thyroxin (T4) favor Schistosoma mansoni development. Their effect is similar, rather than identical; moreover, cotreatment acts synergistically on parasites. This questioned a common mediator to their action, which we hypothesized was host glucose metabolism. Infection with S. mansoni resulted in an early peak in glycemia immediately followed by a peak of insulinemia (D7-21). In IL-7 + T4 cotreated infected animals, the peak of insulin was abrogated. We further assessed the consequences of experimentally induced glucose- or insulin-level variations on parasite development. Insulin treatment from day 14 to day 21 post-infection (PI) led to increased worm burden and parasite size, thus mimicking the effect of T4 on schistosome development. Interestingly, insulin treatment did not modify glycemia yet abrogated the hyperinsulinemia, normally occurring during infection. Finally, these treatments were associated with an alteration of the expression of parasite genes involved in glucose uptake. These experiments characterize the elaborate links between parasite and host metabolism and their reciprocal influences.Journal of Parasitology 09/2005; 91(4):737-44. · 1.26 Impact Factor - [Show abstract] [Hide abstract]
ABSTRACT: The epidermal growth factor receptor (EGF-R) plays an important role in development and cell differentiation, and homologues of EGF-R have been identified in a broad range of vertebrate and invertebrate organisms. This work concerns the functional characterization of SER, the EGF-R-like molecule previously identified in the helminth parasite Schistosoma mansoni. Transactivation assays performed in epithelial Madin-Darby canine kidney cells co-transfected with SER and a Ras-responsive reporter vector indicated that SER was able to trigger a Ras/ERK pathway in response to human epidermal growth factor (EGF). These results were confirmed in Xenopus oocytes showing that human EGF induced meiosis reinitiation characterized by germinal vesicle breakdown in SER-expressing oocytes. Germinal vesicle breakdown induced by EGF was dependent on receptor kinase activity and shown to be associated with phosphorylation of SER and of downstream ERK proteins. (125)I-EGF binding experiments performed on SER-expressing oocytes revealed high affinity (2.9 x 10(-9) M) of the schistosome receptor for human EGF. Phosphorylation of the native SER protein present in S. mansoni membranes was also shown to occur upon binding of human EGF. These data demonstrate the ability of the SER schistosome receptor to be activated by vertebrate EGF ligands as well as to activate the classical ERK pathway downstream, indicating the conservation of EGF-R function in S. mansoni. Moreover, human EGF was shown to increase protein and DNA synthesis as well as protein phosphorylation in parasites, supporting the hypothesis that host EGF could regulate schistosome development. The possible role of SER as a receptor for host EGF peptides and its implication in host-parasite signaling and parasite development are discussed.Journal of Biological Chemistry 10/2004; 279(36):37407-14. · 4.60 Impact Factor
Publication Stats
396 | Citations | |
100.91 | Total Impact Points | |
Top Journals
Institut de Biologie de Lille
Lille, Nord-Pas-de-Calais, France
University of Lille Nord de France
Lille, Nord-Pas-de-Calais, France
Institut Pasteur de Lille
Lille, Nord-Pas-de-Calais, France
Unité Inserm U1077
Caen, Lower Normandy, France


text:
items tagged with Rozz Tox
Written By: Jeff Ignatius
Section: Music
Category: Feature Stories
2012-09-12 11:11:39
When I spoke by phone with Plume Giant's Nolan Green last week, the interview was scheduled for 10 a.m. in New York, where the folk-ish trio is based. I can't remember the last morning interview I had with a pop, rock, or indie musician, as those breeds tend to shy away from morning engagements. So what self-respecting musician is up at that hour?
"I'm ... currently booking a release tour and working like crazy on the PR for it," Green said. "I was actually already at a different meeting this morning, at 8 a.m."
This detail is not necessarily important or telling, but it illustrates that these May graduates from Yale are actively charting their course, including setting specific goals for sales. So while you probably haven't heard of the band – unless you attended its December show at Rozz-Tox, to which they'll be returning on September 21 – its members are working to change that.
Read More About Open To Discovery: Plume Giant, September 21 At Rozz-Tox...
Written By: Jeff Ignatius
Section: Music
Category: Feature Stories
2012-08-29 14:34:23
The first thing to stress about Hello Quad Cities – Volume 1 is that as compilations go, it's strong from front to back and varied without feeling scattershot. The challenging format tends to result in well-intentioned hodgepodges of second-rate leftovers, but the tracks here – from 12 area bands – are all exclusive, and most were written specifically for the compilation. More importantly, while you might not find all of them to your liking, there isn't a weak link.
The second thing to emphasize is that if you're curious about the project, you shouldn't dawdle. The release is available only on vinyl, and a mere 350 copies were pressed. (Each album includes a download code, but there will be no separate digital or CD release.) And they'll only be sold at a pair of record-release shows, by the featured bands, and at Ragged Records.
Read More About Setting The Scene: "Hello Quad Cities – Volume 1"...
Written By: Jeff Ignatius
Section: Music
Category: Feature Stories
2012-08-29 11:51:07
If you haven't heard of the instrumental duo Blues Control, as an introduction let me try to describe the first two tracks from its Valley Tangents album, which was released in June.
"Love's a Rondo" is a jazzy, piano-based tune with one of the keyboard lines often matched by a fuzzy guitar whose frayed edges serve as a gentle contrast. The rhythms are laid-back and slightly exotic, and there's the feel of unhurried, purposeful improv.
"Iron Pigs" starts with beats followed by majestic, cheesy keyboards followed by scratchy, aggressive noise followed by a piano played on the left side. When it emerges, the lead guitar is expressive yet concise, and memories of that agitated opening quickly melt away.
The band will perform at Rozz-Tox on September 9, and, in an interview earlier this month, Lea Cho described its sound as "instrumental psych rock."
That's as brief a description as you'll get, but it's probably more instructive to repeat some of the more verbose attempts. TinyMixTapes.com wrote that Cho and Russ Waterhouse were "an anomaly to me for ages, and listening to their records only made things worse. Their particular mysticality is created with a deeply abstracted series of layers that end up feeling sublimely confounding alongside the various swoons and gritty feelings of transcendence ... ."
Read More About Elusive But Enticing: Blues Control, September 9 At Rozz-Tox...
Written By: Jeff Ignatius
Section: Music
Category: Feature Stories
2012-08-13 14:09:06
The Facebook biography of the Chicago-based trio Cains & Abels is four words: "honest rock and roll."
That might sound glib, vague, evasive, or even a dig at other bands – and it is. But a truer explanation is that singer/songwriter/bassist David Sampson means it, and to expand on the idea would simply take too long. When I asked him a general question about the genesis of "Money" – from the band's gorgeously, patiently articulated My Life Is Easy album – he talked for more than four minutes.
He touched on how his fictional songs seemed to bring their specific sadnesses into his life, and how he decided – almost as a joke – to write happy songs to conjure a different vibe.
"One of the main troubles in my life is money," he said. He discussed how hip-hop artists rap about what they aspire to, and "if it works out, ... they've made it happen by talking about it. ... So I decided at one point that I should try to write some songs about how awesome it is to be wealthy, or at least comfortable financially."
He then deflated what had seemed a hopeful tale. "I ended up writing a song addressing money as a lover that spurned me," he said. "It didn't actually come out the way I intended it to."
Even Sampson's fantasies are weighed down by truth; he couldn't complete a tongue-in-cheek exercise in wish fulfillment.
Read More About Honestly Alluring: Cains & Abels, August 19 At Rozz-Tox...
Written By: Jeff Ignatius
Section: News/Features
Category: Literature
2012-03-08 14:52:49
Philosophy wouldn't seem to lead naturally to poetry, but it can if you find the right philosopher. For Cincinnati-based poet Matt Hart – who will be reading from his work on Saturday at Rozz-Tox along with poets from the Quad Cities edition of the national journal Locuspoint – it was the 20th Century Austrian philosopher Ludwig Wittgenstein.
Hart fell in love with poetry as an undergraduate at Ball State University, but he studied philosophy. Pursing a graduate degree in the subject at Ohio University, though, "I really bought Wittgenstein hook, line, and sinker. As a result, I quit doing philosophy. One of his main ideas is that philosophy is a sort of mental illness; if you understand him, you quit doing it."
And Wittgenstein offered an alternative to philosophy's relentless rational argument, writing that "philosophy ought really to be written only as a form of poetry."
Read More About Destroy The Language: Matt Hart And The Poets Of "Locuspoint: Quad Cities," March 10 At Rozz-Tox...
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