bluuebunny/crossref_metadata_2025_split huggingface dataset:
Synthesis and Purification of Digoxigenin-Labeled RNA Probes for In Situ Hybridization
Authors: Sive Hazel L. , Grainger Robert M. , Harland Richard M. 
Published: August 2007

INTRODUCTIONIn situ hybridization is the most versatile method for determining when and where embryonic transcripts are expressed. Although a detailed analysis of gene expression often requires analysis of sectioned material, the whole-mount approach is invariably the first method used to localize gene expression. Whole-mount in situ hybridization has also become an invaluable tool for analyzing experimentally manipulated embryos and explants. The information gained using this technique is similar to that obtained from immunohistochemistry and includes not only the location of expression but a semiquantitative estimate of the relative levels of gene expression in different parts of the embryo. Because most genes are first analyzed as genomic or cDNA clones, it is straightforward to derive specific probes from these sequences. In this protocol, standard RNA synthesis using a bacteriophage polymerase is carried out incorporating a digoxigenin-substituted ribonucleotide, dig-UTP. In vitro transcription in the direction opposite to in vivo transcription of the mRNA makes an antisense RNA that is complementary to the mRNA. The ratio of dig-UTP to UTP has been optimized to give efficient synthesis and efficient detection of the probe. The three commercially available bacteriophage polymerases (SP6, T7, and T3) all incorporate the substituted nucleotide with equivalent efficiencies, and thus there is a wide choice of plasmid vectors to prepare the template.
Production of Endoribonuclease-Prepared Short Interfering RNAs (esiRNAs) for Specific and Effective Gene Silencing in Mammalian Cells
Authors: Heninger Anne-Kristin , Buchholz Frank 
Published: August 2007

INTRODUCTIONThe mechanism of RNA interference has emerged as a practical tool to study loss of function in many organisms. To make the method of gene knockdown suitable for mammalian cells, short interfering double-stranded RNAs (siRNAs) need to be applied. This protocol describes a straightforward, low-cost method, which uses recombinant Escherichia coli RNase III to digest long dsRNA into endoribonuclease-prepared short interfering RNAs (esiRNAs). Advantages of this technology are the high efficiency and specificity of the resulting esiRNA and its usefulness for not only small-scale applications, but also high-throughput loss-of-function analyses. Another great asset of esiRNA is its flexibility in design, using Web-based tools such as DEQOR, or predesigned esiRNA sequences from the database RiDDLE. PCR products flanked with T7 promoter sequences are generated, transcribed, and annealed. The resulting long dsRNA is enzymatically digested into a pool of overlapping esiRNAs, which are subsequently spin-column-purified.
Identification of Protein-Protein Interactions with Glutathione-S-Transferase (GST) Fusion Proteins
Authors: Einarson Margret B. , Pugacheva Elena N. , Orlinick Jason R. 
Published: August 2007

INTRODUCTIONGlutathione-S-transferase (GST) fusion proteins have had a wide range of applications since their introduction as tools for synthesis of recombinant proteins in bacteria. GST was originally selected as a fusion moiety because of several desirable properties. First and foremost, when expressed in bacteria alone, or as a fusion, GST is not sequestered in inclusion bodies (in contrast to previous fusion protein systems). Second, GST can be affinity-purified without denaturation because it binds to immobilized glutathione, which provides the basis for simple purification. Consequently, GST fusion proteins are routinely used for antibody generation and purification, protein-protein interaction studies, and biochemical analysis. This article describes the use of GST fusion proteins as probes for the identification of protein-protein interactions.
DrosophilaCell Culture and Transformation: Figure 1.
Authors: Cherbas Lucy , Cherbas Peter 
Published: August 2007

INTRODUCTIONPermanentDrosophilacell lines derived from mixed embryonic tissues including the most commonly used lines, S2 and Kc, have been available for ~30 yr. More recently, lines derived from specific tissues, imaginal discs, and the larval central nervous system have come into use. Although cultured cells were originally used by Drosophilists mainly as convenient sources of DNA or carrier RNA, that situation has changed, as an armamentarium of techniques for using the cells has slowly but steadily evolved. Most investigators useDrosophilacell lines as hosts for transformation experiments. The goal may be to characterize a promoter, to investigate the role of a transcription factor, to overexpress a polypeptide, or to do something more novel. This article provides an organized collection of pointers to published protocols.
The reliability, stability, and predictive utility of the self‐report version of the Antisocial Process Screening Device
Authors: MUÑOZ LUNA C. , FRICK PAUL J. 
Published: August 2007

The psychometric properties of the self‐report version of the Antisocial Processes Screening Device (APSD; Frick & Hare, 2001), a rating scale developed to assess traits associated with the construct of psychopathy in youth, was tested in a sample of 91 non‐referred young adolescents with an average age of 13.38 (SD = 1.75) at the initial assessment. The sample was recruited from a large community‐wide screening, where youth with conduct problems and youth high on psychopathic traits were over‐sampled. The sample was reassessed three times at yearly intervals. The self‐report scores on the APSD showed moderate correlations with parent ratings of psychopathic traits, were moderately stable across 1–2 years, and showed significant correlations with measures of antisocial behavior both concurrently and predictively. One major weakness of the self‐report ratings was the low internal consistency of the subscales, which were much lower than the internal consistency found on the parent report version of the scale.
Parents’ estimations of their own intelligence and that of their children: A comparison between English and Icelandic parents
Authors: FURNHAM ADRIAN , VALGEIRSSON HALLDOR 
Published: August 2007

In this study, 158 parents (79 fathers and 79 mothers) with a mean age of 38.3 yrs (SD = 8.2), estimated their own, and their children's, overall intelligence as well as their children's score on the 12 intelligence scales from the Wechsler's Intelligence Scale for Children (WISC‐III). The sample included English (n = 122) and Icelandic parents (n = 36), and a comparison between them showed few differences except that Icelandic parents’ estimates were lower than English parents’ estimates. The results showed fathers estimated their own overall intelligence higher than mothers estimated theirs and sons were estimated higher than daughters on overall intelligence. Two factors (verbal, performance) of intelligence were identified through factor analysis of the ratings of the 12 WISC subscale score estimates. A hierarchical regression showed that these two factors explained most of the variance in the estimation of the child's overall intelligence; however, gender of child and parents’ self‐estimated own overall intelligence added incremental variance.
Multiprotein genetic vaccine in the SIV‐Macaca animal model: a promising approach to generate sterilizing immunity to HIV infection
Authors: Maggiorella Maria Teresa , Sernicola Leonardo , Crostarosa Federica , Belli Roberto , Pavone‐Cossut Maria Rosaria , Macchia Iole , Farcomeni Stefania , Tenner‐Racz Klara , Racz Paul , Ensoli Barbara , Titti Fausto 
Published: August 2007

AbstractBackground  Vaccine combining structural and regulatory proteins is an emerging approach to develop an HIV/AIDS vaccine and therefore, the immunogenicity and efficacy of two regimens of immunization combining structural (Gag/Pol, Env) and regulatory (Rev, Tat, Nef) Simian immunodeficiency virus (SIV) proteins were compared in cynomolgus monkeys.Methods  Monkeys were immunized with Modified Vaccine Ankara vector (MVA‐J5) (protocol 1) or with DNA, Semliki forest virus and MVA vectors (DNA/SFV/MVA) (protocol 2). At week 32, all monkeys were challenge intravenously (protocol 1) or intrarectally (protocol 2) with 50 MID50 of SIVmac251. Humoral, proliferative responses and in particular in protocol 2, the frequency of IFN‐γ producing cells, were measured in all monkeys before and after the challenge.Results  Both vaccine regimens elicited humoral and proliferative responses but failed to induce neutralizing antibodies. Upon intravenous challenge, two out of three MVA‐J5 vaccinated monkeys exhibited a long‐term control of the viral replication whereas DNA/SFV/MVA vaccine abrogated the virus replication up to undetectable level in three out of four vaccinated monkeys. A major contribution to this vaccine effect appeared to be the IFN‐γ/ELISPOT responses to vaccine antigens (Gag, Rev Tat and Nef).Conclusions  These results, indicate that multiprotein heterologous prime‐boost vaccination can induce a robust vaccine‐induced immunity able to abrogate virus replication.
Immunogenicity of a DNA prime and recombinant adenovirus boost regime significantly varies between rhesus macaques of Chinese and Indian origins
Authors: Stahl‐Hennig Christiane , Suh You Suk , Park Ki Seok , Sauermann Ulrike , Kim Kwang Soon , Ahn Soshin , Franz Monika , Schulte Reiner , Stolte‐Leeb Nicole , Hunsmann Gerhard , Sung Young Chul 
Published: August 2007

AbstractBackground Due to an ever increasing shortage of rhesus macaques of Indian origin (InR) that have been generally used for preclinical AIDS vaccine trials in non‐human primates, demand is rising for Chinese rhesus macaques (ChR). However, the immunogenicity of an AIDS vaccine candidate has not been compared in parallel in both rhesus macaque subspecies.Methods ChR and InR were immunized with SIV/HIV DNA and adenovirus vaccine and their immune responses to SIV and HIV evaluated.Results SIV Gag‐ and Env‐specific T‐cell responses and SIV‐specific lymphoproliferative responses measured in ChR were significantly weaker than those in InR (P < 0.05). By contrast, antibody responses to SIV Env, Tat, and Nef in ChR were stronger than those in InR (P < 0.05).Conclusions Immunogenicity of an AIDS vaccine can vary significantly depending on the geographic origin implying genetic differences of macaques. This must be considered when describing and interpreting results of such vaccine studies.
Anti‐CD3/28 mediated expansion of macaque CD4+ T cells is polyclonal and provides extended survival after adoptive transfer
Authors: Onlamoon Nattawat , Plagman Nicholas , Rogers Kenneth A , Mayne Ann E. , Bostik Pavel , Pattanapanyasat Kovit , Ansari Aftab A. , Villinger François 
Published: August 2007

Abstract Background  Our lab has previously shown that adoptive transfer of in vitro expanded autologous purified polyclonal CD4+ T cells using anti‐CD3/CD28 coated beads induced antiviral responses capable of controlling simian immunodeficiency virus (SIV) replication in vivo.Results  Expansion on anti‐CD3/28 coated beads was found to induce a true polyclonal expansion as CFSE labeled cells uniformly showed dilution of the dye over several days of culture, in contrast to aliquots of the same cells subjected to mitogen stimulation. Of interest was the finding that CD4+ T cells collected before and during early chronic SIV infection or AIDS stage did not show any or only modest differences in proliferative response or expansion kinetics. The reason for such excellent expansion properties was analyzed by the quantitation of telomerase activity in aliquots of expanding CD4+ T cells from sample collected at various times post‐infection. First, anti‐CD3/28 expanded CD4+ T cells exhibited telomerase levels 2‐ to 20‐fold higher than the starting population of CD4+ T cells. Moreover, while telomerase activity in ex vivo tested CD4+ T cells was found to decrease following SIV infection and disease progression, anti‐CD3/28 expansion appeared to restore significant levels of telomerase activity as no difference was noted in telomerase expression between CD4+ T cells expanded from samples collected before or during the chronic SIV infection. When such expanded and CFSE labeled T cells were autologously transferred to monkeys, evidence for extended survival in vivo was provided as CFSE labeled cells were detected to relatively high levels in blood and spleen at 1 week post‐infection.Conclusion  In summary, the data suggest that anti‐CD3/28 mediated expansion of CD4+ T cells retains its immunotherapeutic potential not only during the early stages of lentiviral infection but also at more advanced stages of disease.
Simian immunodeficiency virus infection induces severe loss of intestinal central memory T cells which impairs CD4+ T‐cell restoration during antiretroviral therapy
Authors: Verhoeven D. , Sankaran S. , Dandekar S. 
Published: August 2007

AbstractBackground  Simian immunodeficiency virus (SIV) infection leads to severe loss of intestinal CD4+ T cells and, as compared to peripheral blood, restoration of these cells is slow during antiretroviral therapy (ART). Mechanisms for this delay have not been examined in context of which specific CD4+ memory subsets or lost and fail to regenerate during ART.Methods  Fifteen rhesus macaques were infected with SIV, five of which received ART (FTC/PMPA) for 30 weeks. Viral loads were measured by real‐time PCR. Flow cytometric analysis determined changes in T‐cell subsets and their proliferative state.Results  Changes in proliferative CD4+ memory subsets during infection accelerated their depletion. This reduced the central memory CD4+ T‐cell pool and contributed to slow CD4+ T‐cell restoration during ART.Conclusion  There was a lack of restoration of the CD4+ central memory and effector memory T‐cell subsets in gut‐associated lymphoid tissue during ART, which may contribute to the altered intestinal T‐cell homeostasis in SIV infection.
Depo‐Provera abrogates attenuated lentivirus‐induced protection in male rhesus macaques challenged intravenously with pathogenic SIVmac239
Authors: Genescà Meritxell , Li Jun , Fritts Linda , Chohan Paul , Bost Kristen , Rourke Tracy , Blozis Shelley A. , McChesney Michael B. , Miller Christopher J. 
Published: August 2007

AbstractBackground  Progesterone administration prior to intravaginal challenge with pathogenic SIVmac239 decreases the protective efficacy of live attenuated vaccines in rhesus macaques.Methods  To determine if progesterone alters the efficacy of live attenuated vaccines through local or systemic effects, seven male rhesus macaques were immunized with SHIV89.6 and then challenged intravenously with SIVmac239. Three of these animals were treated with Depo‐Provera 30 days prior to the SIV challenge.Results  The SHIV animals had significantly lower plasma viral RNA levels than the unimmunized control monkeys, but the Depo‐Provera treated, SHIV‐immunized animals did not. Despite the lack of protection, the Depo‐Provera SHIV animals had strong SIV specific T‐cell responses. However, altered patterns of NK frequency and CD38 T‐cell expression prior to SIV challenge were observed in Depo‐Provera SHIV animals.Conclusions  Depo‐Provera eliminates live‐attenuated lentivirus vaccine efficacy in male rhesus monkeys through systemic effects on antiviral immunity and/or viral replication.
Did Jesus Christ Claim to be God?
Authors: McNabb Vincent 
Published: August 2007

We may imitate the method of our master St. Thomas by setting down the opinion from which we differ. Dean Rashdall of Carlisle has given us a full and perhaps final expression of that opinion in his paper read at the Cambridge Conference of Modern Churchmen under the title “Christ as Logos and Son of God.” He writes :“Jesus did not claim divinity for Himself. He may have called Himself, or more probably allowed Himself to be called, the Messiah or Son of God. But never in any critically well-attested sayings is there anything which suggests that His conscious relation to God was other than that of a man towards God. . . . The speeches of the Fourth Gospel where they go beyond the Synoptic conception cannot be regarded as history, valuable as they may be for theology. The doctrine of our Lord’s divinity must be taken to express the Church’s conception of what Jesus is or should be to His followers and to the world, not His own theory about Himself.“Note 2. . . .If we accept the Synoptic discourses as substantially correct (though not, of course, in every detail, for there are considerable discrepancies between them), it is impossible to regard the Johannine discourses as equally accurate reports ; and even in this Gospel few sentences (when taken apart from the Preface, which does not pretend to represent the words of Jesus, and other comments of the Evangelist) imply actual Godhead in the sense of post-Nicene theology. . . . The claim to be the Son of God does not necessarily imply Godhead” (The Modern Churchman, Sept., pp. 278, 279)
Modern Marriage
Authors: O'Donoghue Timothy 
Published: August 2007

What is wrong with modern marriage? A somewhat interesting correspondence upon this question appeared during the summer in a London morning paper, and hundreds of people took part in it, eager to give an answer in the light of their own experiences. It was obviously a popular, and certainly a timely, subject for discussion : popular, since almost every conceivable and inconceivable school of thought (or what passes for thought) was represented in that curious series of letters. It was, too, a timely subject, since something must be very wrong with modern marriage when so many thousands of men and women are hastening to the divorce courts to get their marriages dissolved. Still, it was not, nor was it intended to be, a serious or a useful discussion ; its headlines were meant to provide something catchy and piquant for the holiday-making public, and if they prevented the newspaper sales from going all to pieces during the dead season, the correspondence amply fulfilled its purpose.Yet the whole matter of modern marriage is far more serious than those light-hearted inquisitors killing time by the summer sea suppose. Marriage is no longer the grave, deliberate, and stable settlement it used to be, and, outside the Catholic Church, there are few marriages to-day in which, from the first, divorce is not contemplated as a possible eventuality. Divorce has, in fact, now assumed the proportions of a scandal and of a portent unique in the annals of Christianity. True, it has been a commonplace in America for many years, but nothing American ever seems to surprise us over much : we look upon her as a young and irresponsible country, and it is the acknowledged privilege of youth and irresponsibility to shock its elders with impunity.
Henry Vaughan
Authors: Makepeace John Foster 
Published: August 2007

Among the “metaphysical poets” of the seventeenth century Richard Crashaw, George Herbert, and Henry Vaughan form a group apart. They were writers of purely devotional poetry in an age when licentious love-poems and “epithalamiums”— what Dr. Johnson styled “amorous ditties”—were the prevailing fashion. They were united by the . immense spiritual debt which, as they both acknowledge, Crashaw and Vaughan owed to “holy Herbert.” And their chief poems were written within the space of twenty years, for Herbert wrote “The Temple” in 1632 ; Crashaw, “Steps to the Temple” in 1648 ; and Vaughan, “Silex Scintillans” in 1654.Johnson’s use of the word “metaphysical” in his Essay on Cowley is a curious instance of how a quite inappropriate epithet may fairly describe a certain peculiarity of poetic style. For, although it has no relation whatever to any philosophical matter in the poetry, it is by no means difficult to see what was in the great critic’s mind. An appearance of profundity of thought produced by cryptic language was what he noted, and he used the word “metaphysical” in much the same way as some modern writers use the word “mystical,” without regard to its exact connotation, as a general term for that which is not easily understood because its meaning does not lie upon the surface. Herbert, Crashaw, and Vaughan were thus, to a certain extent, “metaphysical” poets, like Cowley and Donne. Herbert, indeed, explains, and partly justifies, his use of this form in the poem strangely entitled “Jordan”
The Soldan and the Pastrycook of Paris
Authors: Eden Helen Parry 
Published: August 2007

Long ago, in the days of St. Louis, there lived in the city of Paris a rich pastrycook. He had not always been rich ; in fact, he began in a very small way indeed as one of those itinerant rissole-sellers who go about at night with a large basket covered with a white napkin, selling rissoles and wafer-cakes to the poor scholars of the University. The scholars used to lead most of the pastrycooks no end of a dance, stealing their baskets and eating the rissoles and hanging the empty baskets up as trophies under their windows. But this pastrycook, whose name was Master Otelinus, did not lose his basket twice, but kept his eyes open and stuck to business and rose very high indeed in what was only too appropriately termed in those days (as, indeed, it should be in our own) the mystery of a pastrycook.In due time he built a handsome shop in The Great Street not far from the Sorbonne, and here he stood at his open window—whose bottom shutters let down into the street made a fine counter, while the top ones were hooked back for an awning—and sold pies of all sorts and sizes from morning till night. Pies stuffed with pork and eels and wild-fowl and eggs—sometimes good eggs and sometimes bad, sanis et quandoque itn-mundis—and very peppery cheese custard. And his servants paraded the city—for he no longer went out himself—cheating his old enemies, the students, over black and white prunes and cherries and unripe apples and such-like small gear.
Bishop Gore's Belief in God
Authors: McNabb Vincent 
Published: August 2007

John Henry Newman’s Apologia pro vita sua has been epigrammatically described as “the work of a French sceptic controlled by an English don—or a joint product of Voltaire and Bishop Butler.” Bishop Gore’s Belief in God, which might well be called his Apologia pro fide sua, is of the same dual and complementary parenthood as the earlier Apologia. Cardinal and Bishop alike belong to that resolute band of thinkers for whom the highest truths are to be discovered in a balance of probabilities and even of contradictories.The younger apologist is hardly a less challenging personality than the elder. Few of the Tertullians or Origens of our day have had a life so crowded with happenings and doings. An Oxford Doctor of Divinity who identified his economic thought with Socialism, who interwove his theological thought with Rationalism, and who accepted the ecclesiastical mode of Anglo-Catholicism, had colour enough in his life to attract not only the undergraduate imagination of Oxford, but even the adult commercialism of Birming ham. There was no limit to the influence wielded by a man who seemed prepared to give up everything, even a belief or a Bishopric, in his thirst for the truth. Thus sceptics were dogmatic about the power wielded over men by a man who. . . had been very far After where the shadows are.