Methods for the absolute quantification of a membrane protein are described using isotopically labeled or unlabeled synthetic peptides as standards. Synthetic peptides are designed to mimic peptides that are cleaved from target analyte proteins by proteolytic or chemical digestion, and the peptides selected serve as standards for quantification by LC/MS/MS on a triple quadrupole mass spectrometer. The technique is complementary to relative quantification techniques in widespread use by providing absolute quantitation of selected targets with greater sensitivity, dynamic range, and precision. Proteins that are found to be of interest by global proteome searches can be selected as targets for quantitation by the present method. This method has a much shorter analytical cycle time (minutes versus hours for the global proteome experiments), making it well suited for high-throughput environments. The present approach using synthetic peptides as standards, in conjunction with proteolytic or chemical cleavage of target proteins, allows mass spectrometry to be used as a highly selective detector for providing absolute quantification of proteins for which no standards are available. We demonstrate that quantification is simple and reliable for the integral membrane protein rhodopsin with reasonable recoveries for replicate experiments using low-micromolar solutions of rhodopsin from rod outer segments.
Recent studies of queenship have allowed us to look closely at various aspects of the life of a ruler's consort, from patronage to reputation to commemoration, with a focus on the queens who were wives or mothers of kings. This study, however, will focus on Urraca of Leon-Castile (reigned 1109-26) to examine the unusual role of a reigning queen, that is, the king's daughter and heir to his throne, whose position carried a greater dynastic weight than did a queen consort's. 1 The latter, although far from powerless, wielded authority only because she was married to a king or, in the case of regents and widows, was the mother of a king. 2 These women married into the royal line, whereas reigning queens were of the royal line. In the central Middle Ages, reigning queens were a brief anomaly of the twelfth century, a not-altogether-successful experiment from the point of view of their courts and their heirs. Matilda of England (d. 1167), Melisende of Jerusalem (d. 1161), and Urraca of Leon-Castile (d. 1126) inherited their kingdoms from their fathers. 3 All had turbulent reigns, brought on by parallel situations: the kings, lacking legitimate sons, named daughters as heirs to the throne, but pow-I am grateful to James D'Emilio and Judith Golden for their comments on a prior version of this article, and to the anonymous Speculum readers, whose suggestions were most helpful. Much of this study was written with the support of a Getty Postdoctoral Fellowship in Spain in 2002-3. All translations are my own. 1 Theresa M. Vann, "The Theory and Practice of Medieval Castilian Queenship," in Queens, Regents and Potentates, ed. Theresa M. Vann, Women of Power 1 (Dallas, Tex., 1993), pp. 125-47, esp. p. 146, points out that according to the law in thirteenth-century Castile-Leon (both the Especulo and the Siete partidas of Alfonso X), a queen derived honor from her husband the king and exercised power through him: "Her most important role was the preservation of the king's lineage." This is a key difference between a queen consort and a queen regnant: the latter worked to preserve her own lineage. 2 Eleanor of Aquitaine (d. 1204), perhaps the best known of all medieval queens, demonstrates that a queen consort could indeed wield significant power. However, her role differed from that of a queen regnant, and thus her political strategies and patronage were also different. For Elednor's role as patron, see George T.
La iglesia románica de San Isidoro de León tiene un lugar de gran relevancia en la historia del arte por su participación en las más importantes corrientes escultóricas de Francia y España del final del siglo XI y comienzo del XII. Han sido objeto de estudio las portadas de la fachada y el transepto meridionales pero la desaparecida puerta del transepto norte no se llegó a estudiar hasta ahora. Gracias a una campaña de investigación en marzo de 2005 durante la cual aparecieron varios fragmentos de escultura, se puede ofrecer una posible reconstrucción del tímpano románico dentro del conjunto decorativo del hastial norte.Palabras clave: San Isidoro de León; Santiago de Compostela; Maiestas Domini; románico; escultura de portada; tímpano; Alfonso VI (r. 1065-1109); Urraca (r. 1109-1126).The Romanesque church of San Isidoro in León holds a place of great significance in the history of art because of its participation in the most important sculptural trends around the turn of the 12 th century. While the portals of the south facade and transept have been the object of scholarly attention, the lost north transept portal had not been studied until now. Thanks to a research campaign in March of 2005, we can propose a possible reconstruction of the Romanesque tympanum within the decorative scheme of the north transept facade.Key words: San Isidoro de León; Santiago de Compostela; Christ in Majesty; Romanesque; portal sculpture; tympanum; King Alfonso VI (r. 1065-1109); Queen Urraca (r. 1109-1126).La iglesia de San Isidoro de León ( fig. 1) es uno de los hitos del estilo arquitectónico que denominamos románico y aunque su escultura ha sido estudiada a fondo, hasta ahora no se había abordado la cuestión de la decoración original de la fachada norte. Una campaña de trabajo (marzo de 2005) encaminada a limpiar y ordenar el lapidario isidoriano, conformado a lo largo del tiempo, trajo consigo la aparición de varios fragmentos escultóricos que en su momento pudieron
Background and aimsIrish Travellers are a Roman Catholic endogamous minority group believed to originate from the 12th century whose ethnicity was finally recognised by the Irish Government in March 2017. Latest estimates put their numbers at 40 129 on the island of Ireland, 15 000 in the United Kingdom, 6000 on Mainland Europe and 7000 in America. Nomadism and family are core features of their identity. Early and arranged marraige, frequent child bearing and consanguinity are cultural norms.Our aim was to collate current data on morbidity and mortality of Traveller children with a view to planning of services and provision of healthcare for this population in our region.MethodsA Literature and database(s) search was conducted and relevant clinicians contacted in an effort to compile data on the complexity and heterogeneity of the moden Irish Traveller Community to inform appropriate action in the area of Traveller Child Health.ResultsAlmost 50% of the Irish Traveller population are children under 15 years of age. Between 650 and 850 Traveller babies are born on the island of Ireland every year. The infant mortality rate is 3.5 times that of the general population. 10% of Traveller children are dead before their second birthday. Main causes of mortality are accidents, congenital malformations and inherited metabolic disorders. To date 93 different genetic disorders are reported, 82 of which the genetic basis is known. There are 60 founder mutations. The commonest disorders are Galactosemia, Hurler Syndome and I-Cell disease, the carrier frequency for which is 1/11,1/11 and 1/15 respectively.Our study highlights the huge disease burden imposed on these children by the combination of curcumstances of poverty, increased frequency of genetic disorders in consanguineous communities, peripatetic lifestyle,nomadic mindset, superstition, stigma and distrust and provides useful epidemiological information with particular reference to the healthcare needs of minority groups marginalised in our society.
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