Identification of type A and B isolates of Epstein-Barr virus by polymerase chain reaction

A method is described for the identification of type A and type B isolates of Epstein-Barr virus (EBV) by means of the polymerase chain reaction. The use of three pairs of primers specific for genomic sequences coding for the two forms of EBV nuclear antigen (EBNA), 2A and 2B, and for a DNA sequence from the BamZ/BamR region allows the reliable and rapid detection of type A and B viruses in as little as 1000 EBV positive cells

Factors influencing the response of broiler chicken to glycine supplements in low crude protein diets

The increasing demand for meat and other animal products along with the global limitation of arable land for crop production is expected to result in a shortage of protein-rich feedstuff. Furthermore, the excretion of nitrogenous compounds has negative effects on the environment because of the risk of nitrogen (N) leakage into the groundwater. Ammonia emissions from livestock enterprises have been associated with environmentally damaging effects. The biggest determinant of ammonia emissions associated with livestock farming is excretion of N. Excretion of N in part is inevitable but N excretion can be reduced by avoiding excessive intake of feed protein. At present, there is a substantial lack of knowledge about the requirement of nitrogenous nutrients except for essential amino acids (AA) and the influence of these nutrients on animal physiology. As reported in the literature, this has often led to undesirable effects of low crude protein (CP) diets on growth. This partly is due to a deficient glycine (Gly) and serine (Ser) supply in low CP diets. This thesis focused on factors influencing the response to Gly and Ser in low CP broiler feed because growth response to these AA was inconsistent in the literature. In the first study, a meta-analysis was conducted to investigate the response of broilers to dietary levels of Gly equivalents (Glyequi) of Gly and Ser in existing literature. A curvilinear relationship between Glyequi and daily gain (ADG), daily feed intake (ADFI) and feed efficiency (G:F) was found. The impact of dietary Glyequi on ADFI was low, but G:F and ADG varied markedly at different levels of Glyequi. The effect of dietary Glyequi depended on the supply of both methionine (Met) and cysteine (Cys). The G:F and ADG response to dietary Glyequi was different for different Met:(Met+Cys) ratios and Cys concentrations. This was explained by a decreased necessity of conversion of Met to Cys, for which Ser is needed. Adequate concentrations of both Met and Cys probably reduced the necessity of the conversion of Met to Cys. The second study investigated the effect of threonine (Thr) and choline, which are endogenous precursors of Gly, on the response to dietary Glyequi. An increase in Thr concentration reduced the Glyequi concentration required to achieve certain response levels of G:F and ADG. Choline also exerted a considerable effect, but the Glyequi replacing effect of choline was less pronounced than Thr. The observed replacement values of dietary Thr and choline for dietary Glyequi exceeded the possible replacement values calculated by considering endogenous conversion. This likely originated from an excess supply of other essential AA than Thr. If Thr and, to a lower extent, choline limited growth, then excessive intake of other AA had to be catabolized, resulting in an increased need for Glyequi for uric acid formation. It was concluded that further studies should take Glyequi, choline, and Thr together into consideration when determining the requirements for these nutrients. The third study aimed to investigate whether the growth performance and N utilization of broilers are influenced by different proportions of free and peptide-bound AA in diets, and if his influences Glyequi requirements. The hypothesis was that an increased oxidation of free AA leads to an increased ammonia production, which must be detoxified to uric acid in a Gly-dissipating process. 2×2 factorial arrangements were used where one factor was AA from soy protein isolate or from a free AA mix. The other factor was a low and high level of Glyequi. Replacing AA from the soy protein isolate with free AA reduced ADG and G:F, mainly due to reduced ADFI. Reasons for that cannot be identified clearly. The N efficiency on day 21 was not different between the AA sources, possibly due to the lower AA digestibility of the soy protein isolate and higher urinary excretion of nitrogenous substances in the treatments with the AA mix. Thus, availability of AA for protein synthesis after ingestion probably did not limit broiler growth. The ADG of the treatments with the high Glyequi concentration was higher for both AA sources. This increase was due to higher ADFI by broilers in the treatments with soy protein isolate and due to the increased G:F in the treatments with the AA mix. Contrary to the hypothesis, these responses did not give an indication of different utilization of Glyequi for uric acid synthesis. In conclusion, the response of broiler chicken to dietary Glyequi depends on other dietary characteristics, like the concentrations of Cys, Thr, and choline. The information described in this thesis contribute to enable further optimization of the dietary Glyequi concentration as well as the other dietary characteristics influencing the response to Glyequi. This enables reducing the CP concentration in diets without adverse effects on growth and, therefore, diminishing the negative effects of broiler production on the environment.Der steigende Bedarf an tierischen Produkten bei global begrenztem nutzbaren Land für die Pflanzenproduktion bewirkt, dass eine Knappheit an proteinreichen Futtermitteln zu erwarten ist. Zudem bringt die Ausscheidung von stickstoff-(N)-haltigen Substanzen das Risiko von N-Eintrag in das Grundwasser. Ammoniakemissionen von tierhaltenden Betrieben werden mit einer Vielzahl von umweltschädigenden Auswirkungen in Verbindung gebracht. Der bedeutendste Einflussfaktor auf nutztierbedingte Ammoniakemissionen ist die Ausscheidung von N. Diese kann durch das Vermeiden von überschüssiger Proteinaufnahme vermindert werden. Derzeit besteht eine erhebliche Unkenntnis hinsichtlich des Bedarfs an N-haltigen Nährstoffen abgesehen von essentiellen Aminosäuren (AS) und dem Einfluss dieser Nährstoffe auf die Tierphysiologie. Dies hat in vielen Fällen zu nicht wünschenswerten Auswirkungen rohprotein-(XP)-reduzierter Futtermischungen auf die Wachstumsleistung geführt. Ein Mangel an den nichtessentiellen AS Glycin (Gly) und Serin (Ser) im Futter für Masthähnchen wurde als bedeutende Ursache gefunden, wobei die Auswirkungen von Gly und Ser im Futter auf das Wachstum in Studien sehr unterschiedlich war. Das Ziel der vorliegenden Arbeit war daher Einflussfaktoren auf die Auswirkungen von Gly und Ser im Futter für Masthähnchen zu untersuchen. In der ersten Studie wurde eine Metaanalyse durchgeführt um die Auswirkungen der Konzentration an Gly-Äquivalenten (Glyequi) von Gly und Ser im Futter auf das Wachstum von Masthühnern in zuvor veröffentlichter Literatur zu untersuchen. Zwischen der Glyequi-Konzentration und täglichen Zunahmen (TZ), täglicher Futteraufnahme (TFA) und Futtereffizienz (FE) wurde ein kurvilinearer Zusammenhang festgestellt. Die Auswirkung der Glyequi-Konzentration auf die TFA war gering; dagegen waren FE und TZ deutlich beeinflusst. Die Auswirkung der Glyequi-Konzentration hing von der Versorgung mit Methionin (Met) und Cystein (Cys) ab. Die Auswirkung der Glyequi-Konzentration auf das Niveau der TZ und FE war verschieden bei veränderten Verhältnissen von Met zu Met+Cys und Cys-Konzentrationen. Dies kann durch eine verringerte Notwendigkeit der Cys-Synthese aus Met erklärt werden, für die Ser benötigt wird. Die zweite Studie untersuchte den Einfluss von Threonin (Thr) und Cholin, welche endogene Vorstufen von Gly sind, auf die Auswirkungen von Glyequi. Eine Erhöhung der Thr-Konzentration verringerte die benötigte Glyequi-Konzentration um bestimmte Niveaus an TZ und FE zu erreichen. Die Cholin-Konzentration hatte ebenfalls einen erheblichen Einfluss. Allerdings war der Austauschwert von Cholin zu Glyequi geringer als der von Thr. Die festgestellten Austauschwerte von Thr und Cholin für Glyequi überstiegen die durch endogene Gly-Synthese erklärbare Austauschwerte. Sie sind wahrscheinlich durch eine übermäßige Versorgung mit anderen essentiellen AS außer Thr bedingt. Wenn Thr und in Cholin die Wachstumsleistung begrenzten, mussten andere übermäßig aufgenommene AS katabolisiert werden was zu einem höheren Glyequi-Bedarf für die Bildung von Harnsäure führt. Demnach sollten künftige Studien Glyequi, Thr und Cholin zusammen berücksichtigen, wenn Bedarfswerte für diese Nährstoffe ermittelt werden. Die dritte Studie wurde durchgeführt um zu prüfen, ob die Wachstumsleistung und N-Verwertung von Masthähnchen vom Verhältnis von freien und peptidgebundenen AS im Futter abhängt und ob das den Glyequi-Bedarf beeinflusst. Die Hypothese war, dass eine erhöhte Oxidation von freien AS zu einer erhöhten Ammoniakproduktion führt, das bei einem Gly-verbrauchenden Prozess zu Harnsäure entgiftet wird. Der Austausch der AS-Menge von Sojaproteinisolat gegen freie AS verringerte die TZ und FE vor allem durch eine verringerte TFA. Ursachen dafür können nicht eindeutig geklärt werden. Hinsichtlich der N-Effizienz bewirkten die AS-Quellen keinen Unterschied. Folglich begrenzte die Verfügbarkeit von AS für die Proteinsynthese nach der Aufnahme vermutlich das Wachstum nicht. Die TZ bei den Behandlungen mit hoher Glyequi-Konzentration war bei beiden AS-Quellen erhöht. Diese Steigerung war durch eine höhere TFA bei den Behandlungen mit Sojaproteinisolat und durch eine höhere FE bei den Behandlungen mit der freien AS-Mischung bedingt. Entgegen der Hypothese geben diese Ergebnisse keinen Hinweis auf eine unterschiedliche Verwertung von Glyequi für die Harnsäurebildung. Nach dieser Arbeit hängen die Auswirkungen der Glyequi-Konzentration im Futter von der weiteren Futterbeschaffenheit, wie den Konzentrationen an Cys, Thr und Cholin ab. Die beschriebenen Erkenntnisse lassen eine weitere Optimierung der Konzentrationen von Glyequi und weiterer Nährstoffe zu. Dies ermöglicht die XP-Konzentration im Futter ohne nachteilige Auswirkungen auf das Wachstum zu reduzieren und somit die negativen Auswirkungen der Masthähnchenproduktion auf die Umwelt zu verringern

Quantitative expression analysis of HHV-6 cell receptor CD46 on cells of human cord blood, peripheral blood and G-CSF mobilised leukapheresis cells

Human herpesvirus-6 (HHV-6) can infect blood cells and thereby may inhibit hematopoietic stem and progenitor cell expansion and differentiation. In this context, it has been discussed if early progenitor cells can be infected by HHV-6. CD46 was identified as one possible cellular surface receptor for HHV-6. The study presented here had been done to get insight into the susceptibility of various leukocyte subpopulations to HHV-6 (including early hematopoietic progenitors) by determining the amount of CD46 molecules expressed on their surfaces. Human cord blood cells, peripheral blood cells and G-CSF mobilised progenitor cells were analysed by flow cytometry. CD46 molecule number per cell was determined and compared to calibration beads conjugated with known ratio of PE per bead. Highest CD46 expression was detected on B- lymphocytes, whereas T-lymphocytes only showed about half of the amount found on B cells. Hematopoietic progenitors also carried CD46 at intermediate levels. Unexpectedly, CD46 expression on progenitors from G-CSF mobilised leukapheresis products was approximately 20% of that found on comparable cells from untreated cord blood. In conclusion, hematopoietic progenitor cells express CD46 on their surface, thereby fulfilling a basic requirement for the susceptibility of HHV-6 infection

Estudios espectroscópicos de silicatos Contribución a la determinación espectroscópica de aniones silicatos en silicatos de calcio hidratados

Not availableLos Profesores Kronert y Siegert realizan un estudio de silicatos por espectroscopia infrarroja, comenzando por el concepto de pérdida de simetría en los sólidos, a causa de la cual en el espectro IR correspondiente, aparecen bandas de absorción, que en la estructura regular serían prohibidas; y degeneradas se desdoblan. Efectúan el trabajo sobre silicatos cálcicos hidratados, tanto naturales, como sintetizados en el Laboratorio

Züchtung und Agronomie neuartiger, Vicin-armer Ackerbohnen und Einsatz als einheimisches Eiweißfutter

In Deutschland sind Ackerbohnen nach Erbsen und Lupinen gemessen an der Anbaufläche die drittbedeutendste Leguminose. Eine günstige Eigenschaft von Ackerbohnen als Futtermittel ist ein im Vergleich zu Erbsen und Lupinen höherer Rohproteingehalt. Dennoch ist die Akzeptanz von Ackerbohnen in Deutschland als Futtermittel im Vergleich zu Erbsen und Lupinen eher gering. Voraussetzung für eine höhere Akzeptanz von Ackerbohnen als Futtermittel ist, dass der genaue Futterwert von Ackerbohnen bekannt ist. Wichtige Kriterien sind Aminosäurenverdaulichkeit und Umsetzbare Energie (MEN). Zudem enthalten Ackerbohnen antinutritive Inhaltsstoffe wie Vicin/Convicin, Tannine und Phytat, welche die Verwendbarkeit als Futtermittel einschränken. Aus diesen Gründen wurde im Rahmen von Abo-Vici die Variation des Futterwertes von Ackerbohnen bei Legehennen und Einflüsse hierauf bewertet. Die Ergebnisse geben Aufschluss über Niveau und Variation von Aminosäurenverdaulichkeit und MEN. Unterschiede zwischen Sommer- und Winterackerbohnen hingen vom Anbaustandort ab. Phytat-Gehalte in Ackerbohnen wurden als Ursache für die Variation der Aminosäurenverdaulichkeit gefunden, während Tannine eher die Variation der MEN beeinflussten. Es gab keinen Hinweis auf einen Einfluss von Vicin/Convicin auf Aminosäurenverdaulichkeit und MEN. Eine Schätzung der Aminosäurenverdaulichkeit und MEN anhand chemischer Inhaltsstoffe war nicht mit ausreichender Genauigkeit möglich. Ein Entschälen der Bohnen erhöhte die MEN deutlich und steigerte die Verdaulichkeit einiger Aminosäuren. Die Erkenntnisse dieses Projekts tragen zu einem gezielteren Einsatz von Ackerbohnen in Legehennenfutter bei. Dies bietet die Möglichkeit, eine Überversorgung der Tiere mit Aminosäuren und MEN zu vermeiden und gleichzeitig die Gefahr einer unzureichenden Versorgung zu mindern. Dies ist günstig für die Ziele, einheimische Proteinfuttermittel zu fördern, das Tierwohl zu erhöhen und Emissionen aus der Tierhaltung zu verringern

Cytokine profiles of cord and adult blood leukocytes: differences in expression are due to differences in expression and activation of transcription factors

<p>Abstract</p> <p>Background</p> <p>Stem cell transplantation as therapy for hematological disorders is often hampered by severe graft-versus-host-disease. This may be reduced by umbilical cord blood transplantation, an effect that has been attributed to qualitative differences between neonatal and adult T cells. We compared levels of secreted proteins and cytokine mRNA induced in cord blood leukocytes (CBL) and adult blood leukocytes (ABL) by various stimuli.</p> <p>Results</p> <p>While interleukin-2 (IL-2) levels were similar in CBL and ABL, there was less induction of the Th1 cytokine interferon-γ in CBL. Production of the Th2 cytokines IL-4, IL-5, and IL-13 and the hematopoietic cytokine IL-3 was much lower in CBL versus ABL after T-cell receptor-mediated stimulation, whereas production of GM-CSF was comparable in the 2 cell types. The lower levels of Th1 and Th2 cytokines were maintained in CBL during a 4-day time-course study, while after 12 hours IL-3 and GM-CSF reached in CBL levels similar to those in ABL. For all cytokines except IFNγ, the IC₅₀values for inhibition by cyclosporin A were similar in ABL and CBL. In contrast, there was less expression and activation of transcription factors in CBL. Activation of NF-κB by TPA/ionomycin was detected in ABL but not CBL. Furthermore, there was less expression of the Th subset-specific transcription factors T-bet and c-maf in CBL versus ABL, whereas GATA-3 expression was similar. Expression of T-bet and c-maf correlated with expression of the Th1 and Th2 cytokines, respectively. Time course experiments revealed that T-bet expression was stimulated in both cell types, whereas c-maf and GATA-3 were induced only in ABL.</p> <p>Conclusion</p> <p>The diminished capability of CBL to synthesize cytokines is probably due to decreased activation of NF-κB, whereas differences in Th subsets are due to differences in regulation of Th lineage-specific transcriptions factors. We propose that the reduced incidence and severity of GvHD after allogeneic transplantation of umbilical CB cells is due to lesser activation of specific transcription factors and a subsequent reduction in production of certain cytokines.</p

Autoimmunity to Sphingosine-1-Phosphate-Receptors in Systemic Sclerosis and Pulmonary Arterial Hypertension

Context: Pulmonary arterial hypertension (PAH) is a frequent extracutaneous manifestation of systemic sclerosis (SSc). PAH is characterized by increased vasomotor tone, progressive remodeling of pulmonary arteries and arterioles, consequentially increased pulmonary vascular resistance, right heart hypertrophy, and eventually right ventricular failure. Autoimmunity against G-protein coupled receptors (GPCRs) has been implicated in the development of SSc-associated PAH. Sphingosine-1-phosphate (S1P) receptors (S1PR) present a potential, yet so far untested antigen for PAH autoimmunity, given the documented role of S1P/S1PR signaling in PAH pathogenesis. Objective: We hypothesized that S1P receptors (S1PR) may constitute autoantigens in human patients, and that the prevalence of autoantibodies (aAb) to S1PR1, S1PR2 and S1PR3 is elevated in SSc patients and associated with PAH. Methods: For this exploratory study, serum samples from 158 SSc patients, 58 of whom with PAH, along with 333 healthy control subjects were screened for S1PR-aAb. S1PR1-3 were expressed as fusion proteins with luciferase in human embryonic kidney cells and used to establish novel in-vitro assays for detecting and quantifying S1PR-aAb. The fusion proteins were incubated with serum samples, the aAb-S1PR complexes formed were precipitated by protein-A, washed and tested for luciferase activity. Commercial anti-S1PR-antibodies were used to verify specificity of the assays. Results: All three assays showed dose-dependent signal intensities when tested with S1PR-subtype specific commercial antibodies. Natural aAb to each S1PR were detected in healthy controls with a prevalence of <10% each, i.e., 2.7% for S1PR1-aAb, 3.6% for S1PR2-aAb, and 8.3% for S1PR3. The respective prevalence was higher in the cohort of SSc patients without PAH, with 17.1% for S1PR1-aAb, 19.0% for S1PR2-aAb, and 21.5% for S1PR3. In the subgroup of SSc patients with PAH, prevalence of aAb to S1PR2 and S1PR3 was further elevated to 25.9% for S1PR2-aAb, and 27.6% for S1PR3. Notably, the majority of patients with positive S1PR2-aAb (60.7%) or S1PR3-aAb (71.9%) displayed interstitial lung disease. Conclusion: S1PR1-3 can constitute autoantigens in humans, particularly in SSC patients with PAH. The potential pathophysiological significance for the etiology of the disease is currently unknown, but the elevated prevalence of S1PR2-aAb and S1PR3-aAb in SSC patients with PAH merits further mechanistic investigations

Candida dubliniensis candidemia in patients with chemotherapy-induced neutropenia and bone marrow transplantation.

The recently described species Candida dubliniensis has been recovered primarily from superficial oral candidiasis in HIV-infected patients. No clinically documented invasive infections were reported until now in this patient group or in other immunocompromised patients. We report three cases of candidemia due to this newly emerging Candida species in HIV-negative patients with chemotherapy-induced immunosuppression and bone marrow transplantation

SN2014J gamma rays from the 56^56Ni decay chain

R. Diehl, et al., “SN2014J gamma rays from the 56Ni decay chain”, Astronomy & Astrophysics, Vol. 574, January 2015. The version of record is available online at: https://www.aanda.org/articles/aa/abs/2015/02/aa24991-14/aa24991-14.html Reproduced with Permission from Astronomy and Astrophysics, © ESO 2015.Context. The detection and measurement of gamma-ray lines from the decay chain of 56Ni provides unique information about the explosion in supernovae. SN2014J at 3.3 Mpc is a sufficiently-nearby supernova of type Ia so that such measurements have been feasible with the gamma-ray spectrometer SPI on ESA’s INTEGRAL gamma-ray observatory. Aims. The 56Ni freshly produced in the supernova is understood to power the optical light curve, because it emits gamma rays upon its radioactive decay first to 56Co and then to 56Fe. Gamma-ray lines from 56Co decay are expected to become directly visible through the white dwarf material several weeks after the explosion, as they progressively penetrate the overlying material of the supernova envelope, which is diluted as it expands. The lines are expected to be Doppler-shifted or broadened from the kinematics of the 56Ni ejecta. We aim to exploit high-resolution gamma-ray spectroscopy with the SPI spectrometer on INTEGRAL toward constraining the 56Ni distribution and kinematics in this supernova. Methods. We use the observations with the SPI spectrometer on INTEGRAL, together with an improved instrumental background method. Results. We detect the two main lines from 56Co decay at 847 and 1238 keV, which are significantly Doppler-broadened, and at intensities (3.65 ± 1.21) × 10-4 and (2.27 ± 0.69) × 10-4 ph cm-2 s-1, respectively, at their brightness maximum. We measure their rise toward a maximum after about 60–100 days and a decline thereafter. The intensity ratio of the two lines is found to be consistent with expectations from 56Co decay (0.62 ± 0.28 at brightness maximum, the expected ratio is 0.68). We find that the broad lines seen in the late, gamma-ray transparent phase are not representative of the early gamma-ray emission, and notice instead that the emission spectrum is complex and irregular until the supernova is fully transparent to gamma rays, with progressive uncovering of the bulk of 56Ni. We infer that the explosion morphology is not spherically symmetric, both in the distribution of 56Ni and in the unburnt material which occults the 56Co emission. After we compare light curves from different plausible models, the resulting 56Ni mass is determined to be 0.49 ± 0.09 M⊙.Peer reviewe