The Macrophage-Specific Promoter mfap4 Allows Live, Long-Term Analysis of Macrophage Behavior during Mycobacterial Infection in Zebrafish.

Transgenic labeling of innate immune cell lineages within the larval zebrafish allows for real-time, in vivo analyses of microbial pathogenesis within a vertebrate host. To date, labeling of zebrafish macrophages has been relatively limited, with the most specific expression coming from the mpeg1 promoter. However, mpeg1 transcription at both endogenous and transgenic loci becomes attenuated in the presence of intracellular pathogens, including Salmonella typhimurium and Mycobacterium marinum. Here, we describe mfap4 as a macrophage-specific promoter capable of producing transgenic lines in which transgene expression within larval macrophages remains stable throughout several days of infection. Additionally, we have developed a novel macrophage-specific Cre transgenic line under the control of mfap4, enabling macrophage-specific expression using existing floxed transgenic lines. These tools enrich the repertoire of transgenic lines and promoters available for studying zebrafish macrophage dynamics during infection and inflammation and add flexibility to the design of future macrophage-specific transgenic lines.

Gene loss, adaptive evolution and the co-evolution of plumage coloration genes with opsins in birds.

BACKGROUND: The wide range of complex photic systems observed in birds exemplifies one of their key evolutionary adaptions, a well-developed visual system. However, genomic approaches have yet to be used to disentangle the evolutionary mechanisms that govern evolution of avian visual systems. RESULTS: We performed comparative genomic analyses across 48 avian genomes that span extant bird phylogenetic diversity to assess evolutionary changes in the 17 representatives of the opsin gene family and five plumage coloration genes. Our analyses suggest modern birds have maintained a repertoire of up to 15 opsins. Synteny analyses indicate that PARA and PARIE pineal opsins were lost, probably in conjunction with the degeneration of the parietal organ. Eleven of the 15 avian opsins evolved in a non-neutral pattern, confirming the adaptive importance of vision in birds. Visual conopsins sw1, sw2 and lw evolved under negative selection, while the dim-light RH1 photopigment diversified. The evolutionary patterns of sw1 and of violet/ultraviolet sensitivity in birds suggest that avian ancestors had violet-sensitive vision. Additionally, we demonstrate an adaptive association between the RH2 opsin and the MC1R plumage color gene, suggesting that plumage coloration has been photic mediated. At the intra-avian level we observed some unique adaptive patterns. For example, barn owl showed early signs of pseudogenization in RH2, perhaps in response to nocturnal behavior, and penguins had amino acid deletions in RH2 sites responsible for the red shift and retinal binding. These patterns in the barn owl and penguins were convergent with adaptive strategies in nocturnal and aquatic mammals, respectively. CONCLUSIONS: We conclude that birds have evolved diverse opsin adaptations through gene loss, adaptive selection and coevolution with plumage coloration, and that differentiated selective patterns at the species level suggest novel photic pressures to influence evolutionary patterns of more-recent lineages.

Olfactory Receptor Subgenomes Linked with Broad Ecological Adaptations in Sauropsida.

Olfactory receptors (ORs) govern a prime sensory function. Extant birds have distinct olfactory abilities, but the molecular mechanisms underlining diversification and specialization remain mostly unknown. We explored OR diversity in 48 phylogenetic and ecologically diverse birds and 2 reptiles (alligator and green sea turtle). OR subgenomes showed species- and lineage-specific variation related with ecological requirements. Overall 1,953 OR genes were identified in reptiles and 16,503 in birds. The two reptiles had larger OR gene repertoires (989 and 964 genes, respectively) than birds (182-688 genes). Overall, birds had more pseudogenes (7,855) than intact genes (1,944). The alligator had significantly more functional genes than sea turtle, likely because of distinct foraging habits. We found rapid species-specific expansion and positive selection in OR14 (detects hydrophobic compounds) in birds and in OR51 and OR52 (detect hydrophilic compounds) in sea turtle, suggestive of terrestrial and aquatic adaptations, respectively. Ecological partitioning among birds of prey, water birds, land birds, and vocal learners showed that diverse ecological factors determined olfactory ability and influenced corresponding olfactory-receptor subgenome. OR5/8/9 was expanded in predatory birds and alligator, suggesting adaptive specialization for carnivory. OR families 2/13, 51, and 52 were correlated with aquatic adaptations (water birds), OR families 6 and 10 were more pronounced in vocal-learning birds, whereas most specialized land birds had an expanded OR family 14. Olfactory bulb ratio (OBR) and OR gene repertoire were correlated. Birds that forage for prey (carnivores/piscivores) had relatively complex OBR and OR gene repertoires compared with modern birds, including passerines, perhaps due to highly developed cognitive capacities facilitating foraging innovations.

SELECTED ICONIC CHAMBER MUSIC WORKS WITH PIANO FROM THE NINETEENTH AND TWENTIETH CENTURIES

Chamber music repertoire featuring the piano blossomed from the mid-nineteenth through the early twentieth century. The quantity of works increased greatly during this time and the quality of these works reached the highest level. Among the many symbolic works that were composed were sonatas for a single string instrument with piano, piano trios, quartets: and quintets as well as two-piano works and four-hand duets. Being able to study and perform many of these iconic works before I graduated was one of the major goals I set for myself as a collaborative pianist. The abundance of repertoire has made it easy to choose works considered "iconic" for my dissertation's three recitals. Iconic is defined as "very famous or popular, especially being considered to represent particular opinions or a particular time" in the online Cambridge Advanced Leamer's Dictionary & Thesaurus © Cambridge University. The compositions featured in the recitals were composed from 1842 through 1941, including works by Schumann, Brahms, Faure, Rachmaninoff, Ravel, and Lutoslawski. Choosing the repertoire with my fellow performers in mind was an important part of this dissertation. In addition to trying to make balanced programs which include variety, working with different instruments and performers is one of the most fulfilling parts of the musical experience for me as a collaborative pianist. Joining me for the concerts were members of the Aeolus String Quartet (violinist Nicholas Tavani, violinist Rachel Shapiro, violist Greg Luce, and cellist Alan Richardson), pianist Hsiao-Ying Lin (a doctoral student from the Peabody Conservatory), and my colleagues from the Peabody Institute Preparatory Division (faculty violinist Dr. Christian Tremblay and cellist Alicia Ward), and Derek Smith, Associate Principal violist of the Annapolis Symphony Orchestras). The three recitals were performed in the Gildenhom and Ulrich Recital Halls at the University of Maryland, College Park, Maryland. They are recorded on CD and available on compact discs, which can be found in the Digital Repository at the University of Maryland (DRUM).

SAXOPHONE SONATAS 1980-2010

The purpose of this dissertation project identifies contemporary solo saxophone literature, specifically sonatas between the years 1980 and 2010. The overwhelming majority of repertoire written during these thirty years consisted primarily of either multi-movement or through-composed character pieces. By limiting the selected repertoire to sonatas one can still investigate the breadth of the literature that has helped validate the saxophone in the realm of classical music in a format that has seemingly fallen out of favor with composers. The saxophone had developed a unique voice by the middle of the twentieth century in both Europe and in the United States. European composers such as Claude Debussy, Florent Schmidt, Jacques Ibert, Darius Milhaud, Alexander Glazounov, Erwin Schulhoff and Bernard Heiden recognized the potential and beauty of the instrument, while the saxophone had found quite a different niche in vaudeville, jazz, and military bands in the United States. If not for the dynamic performances by concert saxophonist such as Marcel Mule, Sigurd Rascher, Jean-Marie Londeix, Daniel Deffayet, Cecil Lesson, Larry Teal, Eugene Rousseau, Fredrick Hemke and Donald Sinta, the timbral possibilities and technical virtuosity of the saxophone would not have been discovered. The awe inspiring performances by these soloists led to the commissioning of a multitude of works by composers looking to expand the sonic possibilities of this relatively new instrument. Through the 1970's American composers such as Leslie Bassett, Paul Creston, Henry Brant, Robert Muczynski, and Karel Husa were writing significant works for the saxophone, while European composers such as IngolfDahl, Edison Denisov, Alfred Desenclos, Henri Tomasi and Marius Constant were each making their own contributions, all leading to a significant quantity of repertoire that met the quality demands set by the performers. The compositions chosen for this dissertation project were selected after numerous performance, pragmatic, programming and pedagogical considerations were taken into account. The three recitals occurred on: March 7, 2010, December 10, 2010 and May 1, 2011 in either the Gildenhorn Recital Hall or Lecture Hall 2100.

A SURVEY OF FRENCH KEYBOARD MUSIC FROM 1880 TO 1950

French music flourished from the last quarter of the nineteenth century into the early twentieth century, especially in the genres of opera and orchestral music. Although French keyboard music enjoyed less popularity, being overshadowed by these predominant genres, prominent impressionist figures Claude Debussy (1862-1918) and Maurice Ravel (1875-1937) . . brought its revival to the French music. Scholars consider Debussy to be a frontrunner of Impressionism, and his influence had a major impact on subsequent composers. As a result of his popularity, other significant works by French composers seem to be overlooked by pianists and audiences and are not as often performed. Because keyboard works by Debussy and Ravel are a popular performance choice among pianists, I was eager to examine music by other French composers. Through my resea,rch, I found many great works that warranted further study and deserve a place in the keyboard repertoire. This recording project contains works by lesser-known French composers written between the years of 1880 and 1950, namely Emmanuel Chabrier (1841-1894), Gabriel-Urbain Faure (1845-1924), Charles Koechlin (1867-1950), Albert Roussel (1869-1937), Erik Satie (1866-1925), Francis Poulenc (1899-1963), Darius Milhaud (1892-1974), Robert Casadesus (1899-1972) and Henri Dutilleux (b.1916). Since piano repertoire is abundant, it is sometimes difficult to create a performance program. Therefore, it frequently becomes the default to choosing familiar repertoire rather than using the opportunity to expand the repertoire. As a pianist, I feel responsible to search for hidden musical treasures with which pianists and audiences alike are not so well acquainted. This recording project explores nine lesser-known French compositions written between 1880 and 1950. I expect this to be an opportunity to introduce both pianists and audiences to outstanding but unfamiliar works by French composers. This dissertation was recorded on two compact discs in Dekelboum Concert Hall at Clarice Smith Performing Arts Center of the University of Maryland. The recordings are archived in the University of Maryland Library.

Interrogation of individual intratumoral B lymphocytes from lung cancer patients for molecular target discovery.

Intratumoral B lymphocytes are an integral part of the lung tumor microenvironment. Interrogation of the antibodies they express may improve our understanding of the host response to cancer and could be useful in elucidating novel molecular targets. We used two strategies to explore the repertoire of intratumoral B cell antibodies. First, we cloned VH and VL genes from single intratumoral B lymphocytes isolated from one lung tumor, expressed the genes as recombinant mAbs, and used the mAbs to identify the cognate tumor antigens. The Igs derived from intratumoral B cells demonstrated class switching, with a mean VH mutation frequency of 4%. Although there was no evidence for clonal expansion, these data are consistent with antigen-driven somatic hypermutation. Individual recombinant antibodies were polyreactive, although one clone demonstrated preferential immunoreactivity with tropomyosin 4 (TPM4). We found that higher levels of TPM4 antibodies were more common in cancer patients, but measurement of TPM4 antibody levels was not a sensitive test for detecting cancer. Second, in an effort to focus our recombinant antibody expression efforts on those B cells that displayed evidence of clonal expansion driven by antigen stimulation, we performed deep sequencing of the Ig genes of B cells collected from seven different tumors. Deep sequencing demonstrated somatic hypermutation but no dominant clones. These strategies may be useful for the study of B cell antibody expression, although identification of a dominant clone and unique therapeutic targets may require extensive investigation.

Mix it and fix it: functions of composite olfactory signals in ring-tailed lemurs.

Animals communicating via scent often deposit composite signals that incorporate odorants from multiple sources; however, the function of mixing chemical signals remains understudied. We tested both a 'multiple-messages' and a 'fixative' hypothesis of composite olfactory signalling, which, respectively, posit that mixing scents functions to increase information content or prolong signal longevity. Our subjects-adult, male ring-tailed lemurs (Lemur catta)-have a complex scent-marking repertoire, involving volatile antebrachial (A) secretions, deposited pure or after being mixed with a squalene-rich paste exuded from brachial (B) glands. Using behavioural bioassays, we examined recipient responses to odorants collected from conspecific strangers. We concurrently presented pure A, pure B and mixed A + B secretions, in fresh or decayed conditions. Lemurs preferentially responded to mixed over pure secretions, their interest increasing and shifting over time, from sniffing and countermarking fresh mixtures, to licking and countermarking decayed mixtures. Substituting synthetic squalene (S)-a well-known fixative-for B secretions did not replicate prior results: B secretions, which contain additional chemicals that probably encode salient information, were preferred over pure S. Whereas support for the 'multiple-messages' hypothesis underscores the unique contribution from each of an animal's various secretions, support for the 'fixative' hypothesis highlights the synergistic benefits of composite signals.

African-American Composers for the Concert Saxophone: A Look at Three Prolific Composers

African-American composers within the field of classical music have made very profound contributions to the literature. In the field of chamber music, Scott Joplin, William Grant Still, Adolphus Hailstork and other composers illustrious composers have created an established and well-documented body of repertoire for many orchestral wind instruments. The saxophone repertoire, however, has not been developed as fully due to its limited tradition as an orchestral instrument and its prominence in the tradition of jazz and popular music. African-American composers in particular appear to be significantly under-represented within the standard concert saxophone literature. My personal experiences with saxophone repertoire in academic settings, solo recitals, conferences and in surveys of standard repertoire from nationally-recognized saxophone teachers support this assertion. There are many African-American composers who have made substantial contributions to the body of repertoire for the concert saxophone. This dissertation examines the works of three prolific African-American composers for the concert saxophone; Dr. Yusef A. Lateef, Andrew N. White III, and Dr. David N. Baker. All have composed more than five separate works featuring the concert saxophone. This project comprises three recitals, each dedicated to one of the three composers selected for this dissertation. Each recital presented will present their compositions featuring the saxophone as a soloist with various types of accompaniment. The project also includes newly-created piano reductions of Dr. David Baker's works for saxophone and orchestra made collaboratively with Baker and arranger John Leszczynski.

The Legacy of Oboist and Master Teacher, Robert Bloom

Robert Bloom (1908-1994) was legendary in the education and performance world. Often hailed as one of the last performers of the Golden Era of classical music and a favorite of conductors ranging from Stokowski to Stravinsky to Shaw, Bloom was an orchestral oboist and English hornist, oboe soloist, chamber musician, teacher (Eastman, Yale, Hartt, Manhattan School of Music, Juilliard and Philadelphia's University of the Arts), composer, conductor, editor of masterworks of the 18th century, and, as a founding member of the Bach Aria group, a seminal influence in the post-WWII revival of Baroque music in America. In The Robert Bloom Collection and the Art of Robert Bloom CD and video archives, we see what his musical ideals were in 1)18th-century performance practices, 2) writing new music for the instrument and commissioning new works, and 3) and transcribing music for the oboe and English horn. As an oboist, I believe it is important that Bloom's teachings, historical performance practices and ideas for expanding repertoire are propagated. Therefore, the works chosen for this dissertation illustrated this legacy. My recitals included 1) some of Bloom's published 18th-century baroque elaborations (his term for ornamentation), as well Baroque works which I have elaborated, 2) works written by him and by other oboists/composers (Labate, Roseman) as well as a flute/oboe duo that I commissioned by Dr. Marcus Maroney and 3) transcriptions by both Bloom and myself (Bach, Donizetti, Mendelssohn, Mozart, Handel, Schumann and Telemann). In these three dissertation recitals, I hope to have illustrated some of Robert Bloom's lasting contributions and impact on the oboe world, and to have demonstrated the potential for carrying forward this legacy by studying his teaching and emulating his example.

Dance Based Music on Piano

According to the Merriam-Webster dictionary, the definition of dance is “to move your body in a way that goes with the rhythm and style of music that is being played.” As you can see in that definition, these two important ways of expressing human feelings, music and dance, are very closely related. Countless pieces of music have been composed for dance, and are still being composed. It is impossible and useless to count how many kinds of dances exist in the world. Different kinds of dances have been developed according to their purposes, cultures, rhythm and tempo. For this reason, the field of dance-related music necessarily expanded significantly. A great deal of dance music has been written for orchestras, small ensembles, or vocals. Along with them, keyboard music also has a huge repertoire of dance pieces. For example, one of the most famous form in Baroque period was suites. Suites usually include 5 or more dance movements in the same key, such as Minuet, Allemende, Courant, Sarabande, Gigue, Bourree, Gavotte, Passepied, and so on. Nationalistic dances like waltz, polonaise, mazurka, and tarantella, were wonderful sources for composers like Chopin, Brahms, and Tchaikovsky. Dance-based movements were used for Mozart and Beethoven’s piano sonatas, chamber works and concertos. Composers have routinely traveled around the world to collect folk and dance tunes from places they visit. For example, Bartok and Balakirev's pieces that are based on folk dances from where they had traveled became famous and are still thought to be valuable for studying and performing today. For these reasons, it is clear that dance-related music is a very important part of keyboard music. In three dissertation recitals, to expand my performing repertoire and to understand dance-related music deeper, I tried to explore many different styles of dances, and compare interpretations between composers. This program note contains information about each pieces’ composers, related dances, and backgrounds. I hope this will be helpful for a future performer who’s seeking an effective dance based keyboard piece.

Age-related immune reconstitution during highly active antiretroviral therapy in human immunodeficiency virus type 1-infected children.

OBJECTIVES: To evaluate the immune reconstitution in HIV-1-infected children in whom highly active antiretroviral therapy (HAART) controlled viral replication and to assess the existence of a relation between the magnitude of this restoration and age. METHODS: All HIV-1-infected children in whom a new HAART decreased plasma viral load below 400 copies/ml after 3 months of therapy were prospectively enrolled in a study of their immune reconstitution. Viral load, lymphocyte phenotyping, determination of CD4+ and CD8+ T cell receptor repertoires and proliferative responses to mitogens and recall antigens were assessed every 3 months during 1 year. RESULTS: Nineteen children were evaluated. Naive and memory CD4+ percentages were already significantly increased after 3 months of HAART. In contrast to memory CD4+ percentages, naive CD4+ percentages continued to rise until 12 months. Age at baseline was inversely correlated with the magnitude of the rise in naive CD4+ cells after 3, 6 and 9 months of therapy but not after 12 months. Although memory and activated CD8+ cells were already decreasing after 3 months, abnormalities of the CD8 T cell receptor repertoire and activation of CD8+ cells persisted at 1 year. HAART increased the response to mitogens as early as 3 months after starting therapy. CONCLUSIONS: In children the recovery of naive CD4+ cells occurs more rapidly if treatment is started at a younger age, but after 1 year of viral replication control, patients of all ages have achieved the same level of restoration. Markers of chronic activation in CD8+ cells persist after 1 year of HAART.

Toward understanding the essence of post-translational modifications for the Mycobacterium tuberculosis immunoproteome

CD4⁺ T cells are prominent effector cells in controlling Mycobacterium tuberculosis (Mtb) infection but may also contribute to immunopathology. Studies probing the CD4⁺ T cell response from individuals latently infected with Mtb or patients with active tuberculosis using either small or proteome-wide antigen screens so far revealed a multi-antigenic, yet mostly invariable repertoire of immunogenic Mtb proteins. Recent developments in mass spectrometry-based proteomics have highlighted the occurrence of numerous types of post-translational modifications (PTMs) in proteomes of prokaryotes, including Mtb. The well-known PTMs in Mtb are glycosylation, lipidation, or phosphorylation, known regulators of protein function or compartmentalization. Other PTMs include methylation, acetylation, and pupylation, involved in protein stability. While all PTMs add variability to the Mtb proteome, relatively little is understood about their role in the anti-Mtb immune responses. Here,we reviewMtb protein PTMs and methods to assess their role in protective immunity against Mtb. © 2014 van Els, Corbière, Smits, vanGaans-van den Brink, Poelen, Mascart, Meiring and Locht.

Immediate early gene expression in dog thyrocytes in response to growth, proliferation, and differentiation stimuli.

In dog thyroid cells, insulin or IGF-1 induces cell growth and is required for the mitogenic action of TSH through cyclic AMP, of EGF, and of phorbol esters. HGF per se stimulates cell proliferation and is thus the only full mitogenic agent. TSH and cAMP enhance, whereas EGF phorbol esters and HGF repress differentiation expression. In this study, we have investigated for each factor and regulatory cascade of the intermediate step of immediate early gene induction, that is, c-myc, c-jun, jun D, jun B, c-fos, fos B, fra-1, fra-2, and egr1; fra-1 and fra-2 expressions were very low. TSH or forskolin increased the levels of c-myc, jun B, jun D, c-fos, and fos B while decreasing those of c-jun and egr1. Phorbol myristate ester stimulated the expression of all the genes. EGF and HGF stimulated the expression of all the genes except jun D and for EGF fos B. All these effects were obtained in the presence and in the absence of insulin, which shows that insulin is not necessary for the effects of the mitogens on immediate early gene expression. The definition of the repertoire of early immediate genes inductible by the various growth cascades provides a framework for the analysis of gene expression in tumors. (1) Insulin was able to induce all the protooncogenes investigated except fos B. This suggests that fos B could be the factor missing for insulin to induce mitogenesis. (2) No characteristic pattern of immediate early gene expression has been observed for insulin, which induces cell hypertrophy and is permissive for the action of the other growth factors. These effects are therefore not accounted for by a specific immediate early gene expression. On the other hand, insulin clearly enhances the effects of TSH, phorbol ester, and EGF on c-myc, junB, and c-fos expression. This suggests that the effect of insulin on mitogenesis might result from quantitative differences in the transcription complexes formed. (3) c-myc, c-fos, and jun B mRNA induction by all stimulating agents, whether inducing cell hypertrophy, or growth and dedifferentiation, or growth and differentiation, suggests that, although these expressions are not sufficient, they may be necessary for the various growth responses of thyroid cells. (4) The inhibition of c-jun and egr1 mRNA expression, and the marked induction of jun D mRNA appear to be specific features of the TSH cAMP pathway. They might be related to its differentiating action. (5) fos B, which is induced by TSH, forskolin, phorbol ester, and HGF but not by insulin, could be involved in the mitogenic action of the former factors.

Hermeneutics, accreting receptions, hypermedia: A tool for reference versus a tool for instruction

We provide a select overview of tools supporting traditional Jewish learning. Then we go on to discuss our own HyperJoseph/HyperIsaac project in instructional hypermedia. Its application is to teaching, teacher training, and self-instruction in given Bible passages. The treatment of two narratives has been developed thus far. The tool enables an analysis of the text in several respects: linguistic, narratological, etc. Moreover, the Scriptures' focality throughout the cultural history makes this domain of application particularly challenging, in that there is a requirement for the tool to encompass the accretion of receptions in the cultural repertoire, i.e., several layers of textual traditions—either hermeneutic (i.e., interpretive), or appropriations—related to the given core passage, thus including "secondary" texts (i.e., such that are responding or derivative) from as disparate realms as Roman-age and later homiletics, Medieval and later commentaries or supercommentaries, literary appropriations, references to the arts and modern scholarship, etc. in particular, the Midrash (homiletic expansions) is adept at narrative gap filling, so the narratives mushroom at the interstices where the primary text is silent. The genealogy of the project is rooted in Weiss' index of novelist Agnon's writings, which was eventually upgraded into a hypertextual tool, including Agnon's full-text and ancillary materials. Those early tools being intended primarily for reference and research-support in literary studies, the Agnon hypertext system was initially emulated in the conception of HyperJoseph, which is applied to the Joseph story from Genesis. Then, the transition from a tool for reference to an instructional tool required a thorough reconception in an educational perspective, which led to HyperIsaac, on the sacrifice of Isaac, and to a redesign and upgrade of HyperJoseph as patterned after HyperIsaac.