Block & Bridle Annual 1956-1957

Charles Adams (Faculty Advisor), Delbert Kuhlman, John Klingenberg, Ardyce Haring, Harvey Jorgensen, Roy Volzke, Billie Reed, Paul Yeutter Prof. Loeffel, Carolyn Hall, Byron Kort, Larry Paul, Elwin Mosier, Charles Corkle, Kay Robohm, Daniel Stilwell Duane Stokebrand, Gary Briggs, Walt Patterson, Wendell Mousel, Keith Smith, Darrel Zessin, Richard Bonne, Donald Kasbohm, Bruce Skinner Bob Discoe, Doyle Hulme, Jim Smith, Carl Lorenzen, Jay Cook, Gary Berke, Bob Volk, Roger Hild Donald Kuhl, Russell Person, Ray Cada, Ray DeBower, Bob Dannert, Phil Starck, Kay Knudsen, Jerry Brownfield, Allan McClure, Wally Bierman Morris Ochsner, Warren Mitchell, Ed McReynolds, Gerald Dart, Arza Snyder, Mervy Schliefert, Arley Waldo, Tom Hoffman, John Wink, Virgil Gellermann, Duane Neuman

Block & Bridle Annual 1959-1960

President Roger Wehrbein Vice President Ted Klug Secretary George B. O'Neal Treasurer Ralph Hazen Marshal Bud Reece Historian Tom Kraeger Co-Historian John Zauha Ag. Executive Representative Larry Williams Faculty Advisor Dr. E. B. Peo, Jr. George Ahlschwede Richard Hahn Henry Beel Ralph Hazen Gary Briggs Gary Heineman Leslie Cook Max Hauser Richard Eberspacher Buce Jameson Russ Edeal Leon Janovy William Ehresman Alan Jorgensen Rolland Eubanks John Joyner Mickey Evertson Marshall Jurgens Jesse Felker Ron Kahle Mylon Filkins Donald Kavan Richard Frahm Max Keasling Roger French Ronald Kennedy Angus Garey Ted Klug Ed Gates Herb Kraeger Gerald Gogan Tom Kraeger Gerald Goold Fernando Lagos Jay Graf Gerald Lamberson Lloyd Langemeier Ralph Langemeier Gerald Loseke Donald Meiergerd Lowell Minert John Oeltjen George B. O'Neal Don Ormesher Larry Ott Bud Reece Ron Sabatka Keith Smith Ronald Smith Donn Simonson Daryl Starr Galen Stevens Eugene Turdy Ernest Thayer Charles Thompson Jerry Thompson Eli Thomssen William Watkins Allen Trumble Robert Weber Lawrence Turner Dan Wehrbein Reginald Turner Roger Wehrbein Vance Uden Dick White Max Waldo Billy Williams Blair Williams Larry Williams D. Patrick Wright John Zauha

Angularella audubonensis sp. n. (Dilepididae) and Other Cestodes of Cliff Swallows in Colorado

One hundred forty-five cliff swallows, Petrochelidon pyrrhonota, were examined for cestodes in two separate surveys in Weld County, Colorado. Seven species representing four genera were found. Angularella audubonensis sp. n. is differentiated by the small size of the rostellar hooks (8.4-13.5 μm) and the medial relationship of the cirrus pouch to the osmoregulatory canals. Other species collected were Angularella beema, Anonchotaenia globata, Mayhewia ababili, Vitta magniuncinata, Vitta parvirostris and Vitta riparia. Seven new hosts and geographic distribution records were established for Colorado, and six new records were determined for North America.

Cysts Resembling Hydatids in a Chicken

Two large cysts recovered from the abdominal cavity of a domestic chicken superficially resembled hydatid cysts of Echinococcus spp. No protoscolices were present. Microscopic examination of the internal lining of one of the cysts revealed a single cuboidal to columnar, ciliated epithelium, leading to a diagnosis of oviduct cysts. Earlier records of hydatids of fowl are reviewed. Spanish abstract: Reporte de Caso--Quistes en un pollo semejantes a la hidatidosis. Se encontraron dos quistes grandes en la cavidad abdominal de un pollo doméstico. Superficialmente los quistes parecían quistes hidatidícos de Echinococcus spp. No se encontraron escólecis o cabezas de la tenia. El examen microscópico de la pared interna de uno de los quistes mostró un epitelio ciliado, variando de cuboidal a columnar, conduciendo a un diagnóstico de quiste en el oviducto. Se revisan otros casos de hidatidosis en aves.

Characterization of Opsin Gene Alleles Affecting Color Vision in a Wild Population of Titi Monkeys (Callicebus brunneus)

The color vision of most platyrrhine primates is determined by alleles at the polymorphic X-linked locus coding for the opsin responsible for the middle- to long-wavelength (M/L) cone photopigment. Females who are heterozygous at the locus have trichromatic vision, whereas homozygous females and all males are dichromatic. This study characterized the opsin alleles in a wild population of the socially monogamous platyrrhine monkey Callicebus brunneus (the brown titi monkey), a primate that an earlier study suggests may possess an unusual number of alleles at this locus and thus may be a subject of special interest in the study of primate color vision. Direct sequencing of regions of the M/L opsin gene using feces-, blood-, and saliva-derived DNA obtained from 14 individuals yielded evidence for the presence of three functionally distinct alleles, corresponding to the most common M/L photopigment variants inferred from a physiological study of cone spectral sensitivity in captive Callicebus. Am. J. Primatol. 73:189-196, 2011. (C) 2010 Wiley-Liss, Inc.

WATER VAPOR CONDUCTANCE OF THE LOWER CRETACEOUS DINOSAURIAN EGGS FROM SANAGASTA, LA RIOJA, ARGENTINA: PALEOBIOLOGICAL AND PALEOECOLOGICAL IMPLICATIONS FOR SOUTH AMERICAN FAVEOLOOLITHID AND MEGALOOLITHID EGGS

The water vapor conductance (G(H20)) of the neosauropod eggs from the Lower Cretaceous Sanagasta nesting site in La Rioja Province, Argentina, was examined and compared with other Cretaceous Argentinean oological material. The 2900 mgH(2)O/day.Torr G(H2O) of the Sanagasta eggshells confirms an extremely moist nesting environment and supports field observations of dug-out nests in a geothermal setting. The observed thinning of the outer eggshell surface during incubation increases gas conductance and concomitantly decreases eggshell mechanical resistance during the late ontogenetic stages, thus facilitating embryonic development and hatching. The Sanagasta and Entre Rios Province faveoloolithid eggs display the highest and comparable 61120 values and share several morphological and diagenetic characters, indicating comparable nesting strategy in geothermal settings. However, the faveoloolithid Yamintie and La Pampa Province specimens cluster together with lower G(H20) values closer to the megaloolithid eggs. The Gnu) of the megaloolithid egg Megaloolithus patagonicus was reconsidered and new results are now congruent with other reported megaloolithid GH2O values. Additionally, we hypothesize that V-shaped pore canals of M. patagonicus, which upper sections reach only the top third or half eggshell thickness and, a wider section in the middle would not compromise the overall egg mechanical resistance like vertical pores connecting directly the outer to the inner eggshell surfaces. Such pore spatial arrangement and geometry would enhance, as the eggshell thins during incubation, a greater G(H2O), G(O2) and G(CO2) and facilitate embryonic development in high moisture nesting contents. Overall, data suggests that neosauropod nesting and brooding behaviors were dependent on elevated moisture nesting environments.

Giant magnetofossils and hyperthermal events

Magnetotactic bacteria biomineralize magnetic minerals with precisely controlled size, morphology, and stoichiometry. These cosmopolitan bacteria are widely observed in aquatic environments. If preserved after burial, the inorganic remains of magnetotactic bacteria act as magnetofossils that record ancient geomagnetic field variations. They also have potential to provide paleoenvironmental information. In contrast to conventional magnetofossils, giant magnetofossils (most likely produced by eukaryotic organisms) have only been reported once before from Paleocene-Eocene Thermal Maximum (PETM; 55.8 Ma) sediments on the New Jersey coastal plain. Here, using transmission electron microscopic observations, we present evidence for abundant giant magnetofossils, including previously reported elongated prisms and spindles, and new giant bullet-shaped magnetite crystals, in the Southern Ocean near Antarctica, not only during the PETM, but also shortly before and after the PETM. Moreover, we have discovered giant bullet-shaped magnetite crystals from the equatorial Indian Ocean during the Mid-Eocene Climatic Optimum (similar to 40 Ma). Our results indicate a more widespread geographic, environmental, and temporal distribution of giant magnetofossils in the geological record with a link to "hyperthermal" events. Enhanced global weathering during hyperthermals, and expanded suboxic diagenetic environments, probably provided more bioavailable iron that enabled biomineralization of giant magnetofossils. Our micromagnetic modelling indicates the presence of magnetic multi-domain (i.e., not ideal for navigation) and single domain (i.e., ideal for navigation) structures in the giant magnetite particles depending on their size, morphology and spatial arrangement. Different giant magnetite crystal morphologies appear to have had different biological functions, including magnetotaxis and other non-navigational purposes. Our observations suggest that hyperthermals provided ideal conditions for giant magnetofossils, and that these organisms were globally distributed. Much more work is needed to understand the interplay between magnetofossil morphology, climate, nutrient availability, and environmental variability.

DEVELOPMENT OF MICROSATELLITE PRIMERS FOR JATROPHA CURCAS (EUPHORBIACEAE) AND TRANSFERABILITY TO CONGENERS

Premise of the study: Microsatellite primers were developed for Jatropha curcas (Euphorbiaceae), a tree species with large potential for biofuel production, to investigate its natural genetic diversity and mating system to facilitate the establishment of tree improvement and conservation programs. Methods and Results: Using a protocol for genomic library enrichment, 104 clones containing 195 repeat motifs were identified. Primer pairs were developed for 40 microsatellite loci and validated in 41 accessions of J. curcas from six provenances. Nine loci were polymorphic revealing from two to eight alleles per locus, and six primers were able to amplify alleles in the congeners J. podagrica, J. pohliana, and J. gossypifolia, but not in other Euphorbiaceae species, such as Hevea brasiliensis, Manihot esculenta, or Ricinus communis. Conclusions: The primers developed here revealed polymorphic loci that are suitable for genetic diversity and structure, mating system, and gene flow studies in J. curcas, and some congeners.

The First Occurrence in the Fossil Record of an Aquatic Avian Twig-Nest with Phoenicopteriformes Eggs: Evolutionary Implications

Background: We describe the first occurrence in the fossil record of an aquatic avian twig-nest with five eggs in situ (Early Miocene Tudela Formation, Ebro Basin, Spain). Extensive outcrops of this formation reveal autochthonous avian osteological and oological fossils that represent a single taxon identified as a basal phoenicopterid. Although the eggshell structure is definitively phoenicopterid, the characteristics of both the nest and the eggs are similar to those of modern grebes. These observations allow us to address the origin of the disparities between the sister taxa Podicipedidae and Phoenicopteridae crown clades, and traces the evolution of the nesting and reproductive environments for phoenicopteriforms. Methodology/Principal Findings: Multi-disciplinary analyses performed on fossilized vegetation and eggshells from the eggs in the nest and its embedding sediments indicate that this new phoenicopterid thrived under a semi-arid climate in an oligohaline (seasonally mesohaline) shallow endorheic lacustine environment. High-end microcharacterizations including SEM, TEM, and EBSD techniques were pivotal to identifying these phoenicopterid eggshells. Anatomical comparisons of the fossil bones with those of Phoenicopteriformes and Podicipediformes crown clades and extinct palaelodids confirm that this avian fossil assemblage belongs to a new and basal phoenicopterid. Conclusions/Significance: Although the Podicipediformes-Phoenicopteriformes sister group relationship is now well supported, flamingos and grebes exhibit feeding, reproductive, and nesting strategies that diverge significantly. Our multi-disciplinary study is the first to reveal that the phoenicopteriform reproductive behaviour, nesting ecology and nest characteristics derived from grebe-like type strategies to reach the extremely specialized conditions observed in modern flamingo crown groups. Furthermore, our study enables us to map ecological and reproductive characters on the Phoenicopteriformes evolutionary lineage. Our results demonstrate that the nesting paleoenvironments of flamingos were closely linked to the unique ecology of this locality, which is a direct result of special climatic (high evaporitic regime) and geological (fault system) conditions.

Boron site preference in ternary Ta and Nb boron suicides

X-ray single crystal (XSC) and neutron powder diffraction data (NPD) were used to elucidate boron site preference for five ternary phases. Ta3Si1-xBx (x=0.112(4)) crystallizes with the Ti3P-type (space group P4(2)/n) with B-atoms sharing the 8g site with Si atoms. Ta5Si3-x (x=0.03(1); Cr5B3- type) crystallizes with space group 14/mcm, exhibiting a small amount of vacancies on the 4 alpha site. Both, Ta-5(Si1-xBx)(3), X=0.568(3), and Nb-5(Si1-xBx)(3), x=0.59(2), are part of solid solutions of M5Si3 with Cr5B3-type into the ternary M-Si-B systems (M=Nb or Ta) with B replacing Si on the 8h site. The D8(8)-phase in the Nb-Si-B system crystallizes with the Ti5Ga4-type revealing the formula Nb5Si3B1-x (x=0.292(3)) with B partially filling the voids in the 2b site of the Mn5Si3 parent type. (C) 2012 Elsevier Inc. All rights reserved.

Influence of HAART on Alternative Reading Frame Immune Responses over the Course of HIV-1 Infection

Background: Translational errors can result in bypassing of the main viral protein reading frames and the production of alternate reading frame (ARF) or cryptic peptides. Within HIV, there are many such ARFs in both sense and the antisense directions of transcription. These ARFs have the potential to generate immunogenic peptides called cryptic epitopes (CE). Both antiretroviral drug therapy and the immune system exert a mutational pressure on HIV-1. Immune pressure exerted by ARF CD8(+) T cells on the virus has already been observed in vitro. HAART has also been described to select HIV-1 variants for drug escape mutations. Since the mutational pressure exerted on one location of the HIV-1 genome can potentially affect the 3 reading frames, we hypothesized that ARF responses would be affected by this drug pressure in vivo. Methodology/Principal findings: In this study we identified new ARFs derived from sense and antisense transcription of HIV-1. Many of these ARFs are detectable in circulating viral proteins. They are predominantly found in the HIV-1 env nucleotide region. We measured T cell responses to 199 HIV-1 CE encoded within 13 sense and 34 antisense HIV-1 ARFs. We were able to observe that these ARF responses are more frequent and of greater magnitude in chronically infected individuals compared to acutely infected patients, and in patients on HAART, the breadth of ARF responses increased. Conclusions/Significance: These results have implications for vaccine design and unveil the existence of potential new epitopes that could be included as vaccine targets.

Remarkable coordination behavior of alkyl isocyanides toward unsaturated vicinal frustrated P/B Lewis pairs

The conjugated frustrated phosphane/borane Lewis pairs formed by 1,1-carboboration of a substituted diphenylphosphino acetylene, undergo a synergistic 1,1-addition reaction to n-butyl isocyanide with formation of new B-C and P-C bonds to the former isonitrile carbon atom. Using tert-butyl isocyanide dynamic behaviour between the isocyanide-[B] adduct and the 1,1-addition product formation was observed in solution. The different modes of isocyanide binding to the FLPs in the solid state were characterized using X-ray crystal structure analyses and comprehensive 11B and 31P solid-state magicangle- spinning (MAS-) NMR experiments. The free FLP, the Lewis adduct at the borane group, and the cyclic product resulting from isocyanide addition to both reaction centers, can be differentiated via 11B and 31P isotropic chemical shifts, 11B nuclear electric quadrupole coupling constants, isotropic indirect 11B-31P spin-spin coupling constants, and 11B...31P internuclear distances measured by rotational echo double resonance.

Isolierung, Reinigung und Expression zentraler Enzyme der Biosynthese des pflanzlichen Antiarrhythmikums Ajmalin

Zusammenfassung In der vorliegenden Arbeit wurden im Zuge derAjmalinbiosynthese in Rauvolfia serpentina die NADPH2abhängigen Reduktionsschritte des Alkaloids Vomilenin zu 17O Acetylnorajmalin genauer untersucht.Dabei konnte erstmals die exakte Reaktionsreihenfolgeaufgedeckt und die daran beteiligten Enzyme ausPflanzenzellsuspensionskulturen isoliert und aufgereinigtwerden. Die ausgearbeiteten, optimierten Reinigungsprotokolleführten in wenigen Stufen gezielt zu den voneinandergetrennten Reduktase Fraktionen. Durch die Trennung derReduktase Aktivitäten war der Grundstein gelegt, dasZwischenprodukt der Reaktion anzureichern und mitverschiedenen analytischen Verfahren als 2?-(R)-1.2Dihydrovomilenin zu identifizieren. Die daraufhin vergebenenBezeichnungen Vomilenin Reduktase (EC.1.5.1.32) und 1.2Dihydrovomilenin Reduktase (EC.1.3.1.73) zielen auf dasumzusetzende Substrat ab.Für die Vomilenin Reduktase konnte eine 4 stufige Reinigungüber (NH4)2SO4 Fällung, Anionen-austauschchromatographie mitSOURCE 30Q, Hydrophobe Interaktionschromatographie an SOURCE15Phe und Affinitätschromatographie mit 2’,5’ ADP Sepharoseausgearbeitet werden. Hierbei konnte die 1.2Dihydrovomilenin Reduktase schon nach dem erstensäulenchromatogra-phischen Schritt (S 30Q) abgetrenntwerden. Das am Ende der Proteinreinigung angefertigte SDSGel zeigte nur noch 3 Banden, von denen die beiden bei ca.40 und 43 kDa gelegenen Banden mit dem Aktivitätsverlaufder Vomilenin Reduktase korrelierten. Diese wurden einempartiellen Verdau mit der Endoproteinase LysC unterworfen,wobei jeweils 2 Spaltpeptide erhalten werden konnten.Die 1.2 Dihydrovomilenin Reduktase Reinigung umfaßte 6Reinigungsschritte mit (NH4)2SO4-Fällung, SOURCE 30QAnionenaustauschchromatographie, HydroxylapatitChromatographie, 2’,5’ ADP Sepharose-Chromatographie,Anionenaustausch an DEAE Sepharose und abschließen-denAnionenaustausch über MonoQ. Die resultierendeProteinfraktion wies eine ca. 200 fache Anreicherung an 1.2Dihydrovomilenin Reduktase auf. Auch hierbei wurde die nachSDS Gelelek-trophorese als 1.2 Dihydrovomilenin Reduktasebestimmte Proteinbande (bei ca. 48 kDa) sequen-ziert. Eskonnten vier Peptidfragmente erhalten werden, die ebenso wiedie sequenzierten Peptidstücke der 40 und 43 kDa Bande einehohe Homologie zu Oxidoreduktasen, im einzelnen zuCinnamoylalcohol- und Mannitol Dehydrogenasen, aufwiesen.Um die Identität der sequenzierten Proteinbanden zubestätigen, wurde über „reverse genetics“ die jeweilscodierende cDNA eruiert. Dafür wurden - ausgehend von denPeptidstücken der Mikro-sequenzierung - degenerierte Primerentwickelt und über PCR Teilbereiche der cDNA amplifiziert.Diese konnten für eine radioaktive Durchmusterung einerRauvolfia cDNA Bank herangezogen werden. Alternativ botallein die Kenntnis der spezifischen Nukleotidabfolge dieMöglichkeit der Gewinnung von 5’ und 3’ Ende derVollängenklone durch RACE PCR.Nach Abschluß dieser Arbeiten konnten für die 40 und 48 kDaBande je ein Vollängenklon und für die 43 kDa Bande 2Vollängenklone (Isoformen) gefunden werden. SämtlicheVollängenklone besitzen einen offenen Leserahmen, der durchnicht zu translatierende Bereiche am 5’ und 3‘ Endeeingefaßt wird. Um die entsprechenden Proteine produzierenzu können, mußten die dafür codierenden cDNA Bereiche dereinzelnen Klone in ein geeignetes Vektor Wirt-System(Expressionssystem) eingebracht werden.Nach erfolgreicher Umklonierung wurde die Expression durchIPTG Zugabe kontrolliert und Proteinrohextrakte aus denBakterienstämmen isoliert. Als Substrate wurden Vomilenin,das strukturisomere Alkaloid Perakin und aufgrund derHomologien zu Cinnamoylalcohol und Mannitol Dehydrogenasen Zimtaldehyd, Dihydrozimtaldehyd und D(-)Fructose getestet. In allen E. coli Stämmen konnte ein unspezifischesReduktionspotential nachgewiesen werden, ohne daß jedochVomilenin reduziert wurde. Die Testung der 1.2Dihydrovomilenin Reduktase Klone mußte wegen Substratmangelentfallen.Die weitere Charakterisierung der pflanzlichen Enzymeerbrachte eine enorm hohe Substratspezifität mit einer sichauf Rauvolfia beschränkenden taxonomischen Verbreitung.Die Molekulargewichtsbestimmung für die Vomilenin Reduktaseergab nach Größenausschluß-chromatographie an Superdex 75ein Gewicht von etwa 43 kDa. Das ebenfalls über Superdex 75ermittelte Molekulargewicht für die 1.2 DihydrovomileninReduktase lag bei ca. 49.8 kDa. Weiterhin wurde eine Metallionenabhängigkeit für dieVomilenin Reduktase aufgezeigt und die Cofaktorspezifitätsowie die pH und Temperatur Optima für beide Reduktasenbestimmt.

Ortsaufgelöste Speziation von Schwermetallen in geogenen Proben mit Röntgenabsorptionsspektrometrie (my-XAS)

Die Vergesellschaftung und Bindungsform von Arsen in Düngekalk wurde durch chemische und mineralogische Analysen sowie XANES/EXAFS-Messungen untersucht. Die durch-schnittliche As-Konzentration im Düngekalk (70 mg/kg) überschreitet den Grenzwert der DüMV (40 mg/kg). Arsen ist in Mn- (Romanechit) und Fe-Dendriten (Goethit, Ferrihydrit) angereichert. Seine Oxidationsstufe ist jeweils 5+. µ-EXAFS-Untersuchungen ergaben Hin-weise auf zweizähnige und einzähnige mononukleare Durchdringungskomplexe mit Eisen-oxid. Das Mobilisierungsverhalten von Arsen wurde durch sequentielle Extraktion des Dün-gekalks und Mobilisierungsversuche mit wassergesättigtem Boden untersucht. Die Lösung erfolgte vorwiegend im dritten Extraktionsschritt gemeinsam mit kristallinen Eisenoxiden. Unter moderat anoxischen Bedingungen war im Boden keine zusätzliche Mobilisierung von Arsen aus dem Düngekalk nachweisbar. Erhöhte As-Konzentrationen und As3+-Anteile im Porenwasser traten bei niedrigem Eh unabhängig von Kalkzugabe auf. Eine Kopplung des Arsen-Grenzwerts an den Eisenoxidgehalt erscheint sinnvoll. Ein Messaufbau für Mikro-XAS Imaging wurde in Betrieb genommen. Er ermöglicht die si-multane Erfassung einer Probenfläche von 26,6×6,6 mm² wahlweise im Transmissions- oder Fluoreszenzmodus mit der räumlichen Auflösung 52×52 µm² durch eine CCD-Kamera. Zur Datenverarbeitung wurden IDL-Programme sowie die Fernerkundungssoftware ENVI ver-wendet. Die Messergebnisse zeigen weniger Störungen und Rauschen als die Ergebnisse frü-herer Messungen mit einem Prototyp. Die Ergebnisse und Erfahrungen der Messungen liefern Hinweise für die weitere erfolgreiche Nutzung des Messaufbaus.

Cost and mortality prediction using polymerase chain reaction pathogen detection in sepsis: evidence from three observational trials

Delays in adequate antimicrobial treatment contribute to high cost and mortality in sepsis. Polymerase chain reaction (PCR) assays are used alongside conventional cultures to accelerate the identification of microorganisms. We analyze the impact on medical outcomes and healthcare costs if improved adequacy of antimicrobial therapy is achieved by providing immediate coverage after positive PCR reports.