Synthesis of Galactosyl‐Queuosine and Distribution of Hypermodified Q‐Nucleosides in Mouse Tissues

Queuosine (Q) is a hypermodified RNA nucleoside that is found in tRNAHis, tRNAAsn, tRNATyr, and tRNAAsp. It is located at the wobble position of the tRNA anticodon loop, where it can interact with U as well as C bases located at the respective position of the corresponding mRNA codons. In tRNATyr and tRNAAsp of higher eukaryotes, including humans, the Q base is for yet unknown reasons further modified by the addition of a galactose and a mannose sugar, respectively. The reason for this additional modification, and how the sugar modification is orchestrated with Q formation and insertion, is unknown. Here, we report a total synthesis of the hypermodified nucleoside galactosyl‐queuosine (galQ). The availability of the compound enabled us to study the absolute levels of the Q‐family nucleosides in six different organs of newborn and adult mice, and also in human cytosolic tRNA. Our synthesis now paves the way to a more detailed analysis of the biological function of the Q‐nucleoside family

Modulation of the immunoglobulinproduction in rats with cd28-spezific monoclonal antibodies

Einer der wichigsten co-stimulatorischen Rezeptoren auf T-Zellen ist CD28. In Abhängigkeit vom TZR-Signal nimmt CD28 Einfluss auf die Th1/Th2-Differenzierung der Immunantwort. Die rattenspezifischen mAk JJ316 und JJ319 binden an das CD28-Molekül und haben beide ein gleich starkes co-stimulatorisches Potential. Gabe des mitogenen CD28-spezifischen mAkJJ316 - und nicht des konventionellen mAk JJ319 - führt auch ohne TZR-Signal in vitro zu Produktion und Proliferation der Zellen (Direkte Stimulation). In vivo führt Gabe von JJ316 zu einer Erhöhung der Zellzahl in Milz und Lymphknoten mit einem Maximum nach drei Tagen. In vitro führte Behandlung mi mAk JJ316 zu einem Anstieg Th2-spezifischer Immunglobuline. In dieser Arbeit konnte gezeigt werden, dass die Gabes des mitogenen CD28-spezifischen mAk JJ316 im Vergleich mit dem konventionellen CD28-spezifischen mAk JJ319 auch in vivo zu einer Erhöhung der Th2-spezifischen Immunglobuline (IgG1, IgG2a, IgE) bei Brown-Norway- und Lewis-Ratten führt.On of the most important co-stimulating receptors on t-cells is CD28. Depending on the TZR-signal CD28 influences the Th1-/Th2-differentiation of the immune response. The rat-specific mAb JJ316 and JJ319 bind the CD28-molecule and have the same co-stimulatory potential. Treatment with the mitogeneous CD28-specific mAb JJ316 - and not with the conventional mAb JJ319 - leads also without a TZR-signal in vitro to a production and proliferation of cells (direct stimulation). In vivo treatment with JJ316 leads to an increase of the cell number in the spleen an lymphnodes with a maximum rate after three days. In vitro treatment with mAb JJ316 leads to an increase of Th2-specific immunoglobullins. This thesis shows that treatment with mitogeneous CD28-specific mAb JJ316 in comparision with the conventional CD28-specific mAb JJ316 leads also in vivo to an increase of the Th2-specific immunoglobulins (IgG1, IgG2a, IgE) in brown-norway- and lewis-rats

Detection of HPV16 in Esophageal Cancer in a High-Incidence Region of Malawi

This study was designed to explore the role of human papillomavirus (HPV) in esophageal squamous cell carcinoma (ESCC). Fifty-five patients receiving diagnostic upper gastrointestinal endoscopy at Zomba Central Hospital or Queen Elizabeth Hospital in Blantyre (Malawi) in 2010, were included in our study. Formalin-fixed paraffin-embedded biopsies were collected for histopathological diagnosis. HPV DNA was detected using multiplex Quantitative PCR (qPCR) and in situ hybridization (ISH). p16INK4a staining served as a surrogate marker for HPV oncogene activity. Cell proliferation was determined by Ki-67 staining. Human immunodeficiency virus (HIV) status was evaluated by serology. Data on the consumption of alcohol and tobacco, and history of tuberculosis (TBC), oral thrush, and Herpes zoster, were obtained by questionnaire. Forty patients displayed ESCC, three displayed dysplastic epithelium, and 12 displayed normal epithelium. HPV16 was detected in six ESCC specimens and in one dysplastic lesion. Among HPV-positive patients, viral load varied from 0.001 to 2.5 copies per tumor cell. HPV DNA presence could not be confirmed by ISH. p16INK4a positivity correlated with the presence of HPV DNA (p = 0.03). Of particular note is that the Ki-67 proliferation index, in areas with diffuse nuclear or cytoplasmatic p16INK4a staining ≥50%, was significantly higher in HPV-positive tumors compared to the corresponding p16INK4a stained areas of HPV-negative tumors (p = 0.004). HPV infection in ESCC was not associated with the consumption of tobacco or alcohol, but there were significantly more patients drinking locally brewed alcohol among HPV-positive tumor patients compared to non-tumor patients (p = 0.02) and compared to HPV-negative tumor patients (p = 0.047). There was no association between HIV infection, history of TBC, Herpes zoster, oral thrush, or HPV infection, in ESCC patients. Our indirect evidence for viral oncogene activity is restricted to single tumor cell areas, indicative of the role of HPV16 in the development of ESCC. The inhomogeneous presence of the virus within the tumor is reminiscent of the “hit and run” mechanism discussed for β-HPV types, such as HPV38

<i>Helicobacter pylori</i>, HIV and Gastric Hypochlorhydria in the Malawian Population

<div><p>Background</p><p>HIV and <i>Helicobacter pylori</i> are common chronic infections in sub-Saharan Africa. Both conditions can predispose to gastric hypochlorhydria that may be a risk factor for enteric infections and reduced drug absorption. We have investigated to what extent HIV and <i>H</i>. <i>pylori</i> infections are associated with hypochlorhydria in a Malawian cohort of patients undergoing endoscopy.</p><p>Methods</p><p>104 sequential symptomatic adults referred for gastroscopy at Queen Elizabeth Central Hospital, Blantyre, Malawi, had blood taken for rapid HIV testing and fasting serum gastrin analysis. Gastric fluid was aspirated for pH testing, and gastric biopsies were taken.</p><p>Results</p><p>After 9/104 HIV-infected patients who were already established on anti-retroviral therapy were excluded, 17/95 (25.0%) were seropositive for untreated HIV, and 68/95 (71.6%) patients were <i>H</i>. <i>pylori</i> positive by histology. Hypochlorhydria (fasting gastric pH>4.0) was present in 55.8% (53/95) of patients. <i>H</i>. <i>pylori</i> infection was significantly associated with hypochlorhydria (OR 2.91, [1.02-7.75], p=0.046). While single infection with HIV was not significantly independently associated with hypochlorhydria. <i>H</i>. <i>pylori</i> and HIV co-infection was more strongly associated with hypochlorhydria (OR 6.25, [1.33-29.43], p=0.020) than either infection alone, suggesting an additive effect of co-infection. HIV infection was associated with higher serum gastrin levels (91.3pM vs. 53.1pM, p=0.040), while <i>H</i>. <i>pylori</i> infection was not (63.1pM vs. 55.1pM, p=0.610). Irrespective of <i>H</i>. <i>pylori</i> and HIV status, most patients (>90%) exhibited pangastritis. Only three patients had histological evidence of gastric atrophy, of which only one was HIV-infected.</p><p>Conclusion</p><p><i>H</i>. <i>pylori</i> infection was associated with fasting hypochlorhydria, while HIV was not independently associated. HIV and <i>H</i>. <i>pylori</i> co-infection, however, was more strongly associated with hypochlorhydria than <i>H</i>. <i>pylori</i> infection alone. The mechanism of this apparent additive effect between HIV and <i>H</i>. <i>pylori</i> remains unclear, but appears to be related to chronic pangastritis rather than gastric atrophy, and associated with hypergastrinaemia in HIV-infected individuals.</p></div

Univariate and binary logistic regression by factors potentially associated with hypochlorhydria.

<p>Univariate and binary logistic regression by factors potentially associated with hypochlorhydria.</p

The scatterplot displays the serum gastrin (pM) levels for the four groups in relation to <i>H</i>.<i>pylori</i> and HIV infection status.

<p>The scatterplot displays the serum gastrin (pM) levels for the four groups in relation to <i>H</i>.<i>pylori</i> and HIV infection status.</p

Mean gastric pH values by age, gender, <i>H</i>. <i>pylori</i> and HIV status.

<p>Mean gastric pH values by age, gender, <i>H</i>. <i>pylori</i> and HIV status.</p

Findings at gastroscopy; the examination was entirely normal in 27 (26%) patients.

<p>Findings at gastroscopy; the examination was entirely normal in 27 (26%) patients.</p

Histological assessment of antral and corpus (body) biopsies from all subjects.

<p>Histological assessment of antral and corpus (body) biopsies from all subjects.</p