Catalytic properties of 26 S and 20 S proteasomes and radiolabeling of MB1, LMP7, and C7 subunits associated with trypsin-like and chymotrypsin-like activities*

20 and 26 S proteasomes were isolated from rat liver. The procedure developed for the 26 S proteasome resulted in greatly improved yields compared with previously published methods. A comparison of the kinetic properties of 20 and 26 S proteasomes showed significant differences in the kinetic characteristics with certain substrates and differences in the effects of a protein substrate on peptidase activity. Observed differences in the kinetics of peptidylglutamyl peptide hydrolase activity suggest that the 26 S complex cannot undergo the conformational changes of 20 S proteasomes at high concentrations of the substrate benzyloxycarbonyl (Z) -Leu-Leu-Glu-b-naphthylamide. Various inhibitors that differentially affect the trypsin-like and chymotrypsin-like activities have been identified. Ala-Ala-Phe-chloromethyl (CH2Cl) inhibits chymotrypsin-like activity assayed with succinyl (Suc) -Leu-Leu-Val-Tyr-AMC, but surprisingly not hydrolysis of Ala-Ala-Phe-7-amido-4-methylcoumarin (AMC). Tyr-Gly-Arg-CH2Cl inhibits Suc-Leu-Leu-Val-Tyr-AMC hydrolysis as well as trypsinlike activity measured with t-butoxycarbonyl (Boc) -Leu-Ser-Thr-Arg-AMC, while Z-Phe-Gly-Tyr-diazomethyl (CHN2) was found to inhibit only the two chymotrypsin- like activities. Radiolabeled forms of peptidyl chloromethane and peptidyl diazomethane inhibitors, [3H]acetyl-Ala-Ala-Phe-CH2Cl, [3H]acetyland radioiodinated Tyr-Gly-Arg-CH2Cl, and Z-Phe-Gly- Tyr-(125I-CHN2), have been used to identify catalytic components associated with each of the three peptidase activities. In each case, incorporation of the label could be blocked by prior treatment of the proteasomes with known active site-directed inhibitors, calpain inhibitor 1 or 3,4-dichloroisocoumarin. Subunits of labeled proteasomes were separated either by reverse phase-HPLC and SDS-polyacrylamide gel electrophoresis or by twodimensional polyacrylamide gel electrophoresis followed by autoradiography/fluorography and immunoblotting with subunit-specific antibodies. In each case, label was found to be incorporated into subunits C7, MB1, and LMP7 but in different relative amounts depending on the inhibitor used, consistent with the observed effects on the different peptidase activities. The results strongly suggest a relationship between trypsin-like activity and chymotrypsin-like activity. They also help to relate the different subunits of the complex to the assayed multicatalytic endopeptidase activities

Do positional concerns destroy social capital : evidence from 26 countries

Research on the effects of positional concerns on individuals' attitudes and behavior is sorely lacking. To address this deficiency, we use the International Social Survey Programme 1998 data on 25,000 individuals from 26 countries to investigate the impact of relative income position on three facets of social capital: horizontal and vertical trust as well as norm compliance. Testing relative deprivation theory, we identify a deleterious positional income effect for persons below the reference income, particularly for their social trust and confidence in secular institutions. Also often a social capital-lowering effect of relative income advantage occurs, while a rise in absolute income almost always contributes positively. These results indicate that a rise in income inequality in a society too large is rather detrimental to the formation of social capital. (JEL Z130, I300, D310)

The Times They Are A Changin': The Effect of Institutional Change on Cooperative Behaviour at 26,000ft over Sixty Years

In 2006, Sir Edmund Hillary lambasted the modern climbing fraternity for abandoning other climbers to a slow frozen death on Everest, claiming that in his day they would never leave someone to die. This followed the controversial death of David Sharp, passed by an estimated 40 climbers who were more interested in the summit than the life of a fellow human being. But was this stinging criticism true or just the faded recollections of a former climbing giant? This book investigates that claim through a narrative analysis, which combines the empirical analysis of Hawley and Salisbury's Himalayan Expedition Database with the anecdotal evidence provided by a plethora of newspaper articles and books. While there is evidence supporting the claim that commercialization is to blame for the breakdown of pro-social behaviour, the results cannot conclude if it is the commercial climber or the operator driving the problem and that the Sherpa are the saving grace.

Characterization of 26 new microsatellite loci in the dugong (Dugong dugon)

Twenty-six microsatellite loci have been isolated from a dugong (Dugong dugon). The average heterozygosity was 0.52 with two to 10 alleles per locus surveyed from 50 individuals. The markers are suitable for genetic mark-recapture (PID = 5 × 10-16) in dugongs and they could also be used to quantify physical tag loss, estimate relatedness, assign paternity, elucidate population structure and identify migrants.

"En plan för jämställdhet borde kanske utarbetas, men förekommer det ett behov?" : En kartläggning av 26 svenskspråkiga kommuners främjande av jämställdhet

Jämställdhetslagen från 1995 kräver att både myndigheter och arbetsgivare med aktiva och målinriktade åtgärder främjar jämställdheten, och denna pro gradu-avhandling undersöker närmare hur denna lag implementerats i de finlandssvenska kommunerna (kommuner med majoritetsspråk svenska) under en 10årsperiod. I kommunerna jobbar 80% kvinnor, och 38% av alla kvinnliga arbetstagare hittas i arbeten inom den offentliga sektorn. Kommunens skyldighet att trygga dessa kvinnors lika villkor inom kommunorganisationen ingår både som en del i arbetsgivaransvaret, men även som del i statens demokratiska utövande genom myndighetsansvaret. De tre planer som ingår i mitt material innehåller en kartläggning av jämställdhetsområdet i olika omfattning, definierade mål och åtgärder och ingen av planerna har följts upp efter att de godkänts. Planerna varierar både i utformning och omfattning men också i ansvarsspecificering och åtgärdernas konkretisering, vilken ger en varierande kvalitet och därmed även varierande förutsättning att genomdrivas och därmed uppfylla lagens ålägganden. Två av kommunerna har eller har haft en jämställdhetskommitté. I de 26 inkomna svarsformulär görs en närläsning av hur begreppet jämställdhet används och hur jämställdhetslagen tolkas och orsakerna till frånvaron av jämställdhetsåtgärder. På frågan om varför jämställdhetsarbetet inte genomförts helt eller delvis utkristalliserade sig sex olika argumentationsteman: 1. Myten om den jämställda kommunen, innebär en hänvisning till att det inte förekommit några problem med jämställdheten. 2. Den naturligt integrerade jämställdheten, är kommunens policy och sker utan planer, men en liten kritik av problematiken med frånvaron av planer framförs av några kommuner. 3. Könens komplettering, utgår jämställdhetsbegreppet ifrån i några kommuner. 4. Den matematiska jämställdheten, är argument där man genom hänvisningar till kvinnor på olika högre positioner visar på att jämställdheten är uppnådd. 5.En fråga om resurser, används som ett argument till jämställdhetsarbetets frånvaro i kommunen. 6. Avsaknad av intresse, hos personalen och allmänheten är ett annat argument för ogenomförda jämställdhetsåtgärder. Det framkommer tydligt i materialet att kommunerna anser sig följa jämställdhetslagen, samtidigt som det blir tydligt att det råder en oklarhet om vad jämställdhetslagen från 1995 egentligen innehåller. En tydlig sammanblandning mellan den lagstadgade kvoteringen och skyldigheten att aktivt främja jämställdhet träder fram i materialet. Det ofullständiga jämställdhetsarbetet strandar på olika punkter, men en avgörande anser punkt är oförståelsen inför begreppet jämställdhet och därmed också lagens syfte och innehåll. Avhandlingens slutsats är att frånvaron av en könsmaktanalys försvårar förståelsen av ojämställdhetens komplexhet, dvs. dess orsaker, mångskiftande funktion och olika uttryck i organisationens strukturer och i de handlande aktörerna.

The Gottschalk family seated around the dining table; Seelhorst Strasse 26, Hannover Portraits Family

From left to right: Kurt, Therese nee Molling, Ursula, the maid, Elizabeth and Hal,

Tombstone of Arthur Molling; Strangriede Cemetery (R 26.05); Hannover Cemeteries Tombs

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Tombstone of Jeanette Molling nee Benjamin; Strangriede Cemetery (R 26.06); Hannover Cemeteries Tombs

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Detail of tombstone of Arthur Molling; Strangriede Cemetery (R 26.05); Hannover Cemeteries Tombs

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Tombstone of Arthur Molling; Strangriede Cemetery (R 26.05); Hannover Cemeteries Tombs

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Tombstone of Jeanette Molling nee Benjamin; Strangriede Cemetery (R 26.06); Hannover Cemeteries Tombs

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Detail of tombstone of Arthur Molling; Strangriede Cemetery (R 26.05); Hannover Cemeteries Tombs

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Organometallic chemistry of diphosphazanes. Part 26. Ruthenium hydride complexes of chiral and achiral diphosphazane ligands and asymmetric transfer hydrogenation reactions

The half-sandwhich ruthenium chloro complexes bearing chelated diphosphazane ligands, [(eta(5)-Cp)RuCl{kappa(2)-P,P-(RO)(2)PN(Me)P(OR)(2)}] [R = C6H3Me2-2,6] (1) and [(eta(5)-Cp*)RuCl{kappa(2)-P, P-X2PN(R)PYY'}] [R = Me, X = Y = Y' = OC6H5 (2); R = CHMe2, X-2 = C20H12O2, Y = Y' = OC6H5 (3) or OC6H4'Bu-4 (4)] have been prepared by the reaction of CpRu(PPh3)(2)Cl with (RO)(2)PN(Me)P(OR)(2) [R = C6H3Me2-2,6 (L-1)] or by the reaction of [Cp*RuCl2](n) with X2PN(R)PYY' in the presence of zinc dust. Among the four diastereomers (two enantiomeric pairs) possible for the "chiral at metal" complexes 3 and 4, only two diastereomers (one enantiomeric pair) are formed in these reactions. The complexes 1, 2, 4 and [(eta(5)-Cp)RuCl {kappa(2)-P,P-Ph2PN((S)-*CHMePh)PPhY)] [Y = Ph (5) or N2C3HMe2-3,5 (SCSPRRu)-(6)] react with NaOMe to give the corresponding hydride complexes [(eta(5) -Cp)RuH {kappa(2)-P,P-(RO)(2)PN(Me)P(OR)(2)}] (7), [(eta(5)-Cp*)RuH {kappa(2)-P,P'-X2PN(R)PY2)] [R = Me, X = Y = OC6H5 (8); R = CHMe2, X-2 = C20H12O2, Y = OC6H4'Bu-4 (9)] and [(eta(5) -Cp)RuH(kappa(2)-P, P-Ph2PN((S)-*CHMePh)PPhY)][Y =Ph (10) or N2C3HMe2-3,5 (SCSPRRu)(11a) and (SCSPSRu)-(11b)]. Only one enantiomeric pair of the hydride 9 is obtained from the chloro precursor 4 that bears sterically bulky substituents at the phosphorus centers. On the other hand, the optically pure trichiral complex 6 that bears sterically less bulky substituents at the phosphorus gives a mixture of two diastereomers (11a and 11b). Protonation of complex 7 using different acids (HX) gives a mixture of [(eta(5)- Cp)Ru(eta(2)-H-2){kappa(2)-P, P-(RO)(2)PN(Me)P(OR)(2))]X (12a) and [(eta(5)-Cp)Ru(H)(2){kappa(2)-P, P-(RO)(2)PN(Me)P(OR)(2)}]X (12b) of which 12a is the major product independent of the acid used; the dihydrogen nature of 12a is established by T, measurements and also by synthesizing the deuteride analogue 7-D followed by protonation to obtain the D-H isotopomer. Preliminary investigations on asymmetric transfer hydrogenation of 2-acetonaphthone in the presence of a series of chiral diphosphazane ligands show that diphosphazanes in which the phosphorus centers are strong pi-acceptor in character and bear sterically bulky substituents impart moderate levels of enantioselectivity. Attempts to identify the hydride intermediate involved in the asymmetric transfer hydrogenation by a model reaction suggests that a complex of the type, [Ru(H)(Cl){kappa(2)-P,P-X2PN(R)PY2)(solvent)(2)] could be the active species in this transformation. (c) 2007 Elsevier B.V. All rights reserved.