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Biomedical subjects

G Sommer

Publications and source records attributed to G Sommer.

At least 37 records · Page 2Linked to original sources

A new class of defective hepatitis B virus genomes with an internal poly(dA) sequence.

Sequence heterogeneity of hepatitis B virus (HBV) is increasingly recognized to play a role in virus-host interaction. We have used a recently established method for HBV full-length genome amplification to search for novel types of HBV variants and to investigate further the sequence heterogeneity of HBV genome populations. Using this method, a substantial fraction of HBV genomes much shorter than wildtype size was found in some sera and liver biopsies from infected patients. Cloning and sequencing of a number of these HBV genomes as well as hybridization studies revealed a new minor class of HBV genomes with an internal poly(dA) sequence approximately 60 to more than 100 nucleotides long in 4 of 10 patients. The 5'-ends of the internal poly(dA) sequences are located at positions corresponding to the authentic processing/polyadenylation sites of the RNA pregenome, whereas the positions of the 3'-ends are variable due to different sizes of adjacent deletions. These data suggest that the poly(A) tail of the pregenomic RNA is occasionally reverse transcribed by the HBV P-protein and during this process a deletion seems to be introduced into the DNA minus strand. We propose a mechanism by which this could be accomplished during DNA minus strand synthesis.

Base Sequence↗

Heterogeneity and common features of defective hepatitis B virus genomes derived from spliced pregenomic RNA.

Defective hepatitis B virus (HBV) genomes derived from packaging and reverse transcription of spliced RNA pregenomes were reported to be associated with progression to chronic infection. Since only two types with similarly spliced regions were characterized so far we reasoned that additional "spliced" genome variants may exist. Therefore, we isolated a large number of defective HBV genomes from sera of seven chronic carriers by full-length PCR. Forty-eight were found to contain deletions caused by splicing as identified by cloning, subgenomic PCR, and sequencing. In total, 11 types of spliced genomes derived from excision of 10 different introns were present in various combinations in each serum. This diversity resulted from alternative usage of five splice donor and four acceptor sites present in most but not all HBV genotypes. All spliced genomes shared sequence elements essential for replication as well as for transcription of the pre-C and pregenome/C mRNAs and the X mRNA. Moreover, all contained the coding regions for the X protein and for precore/core or precore/ core fusion proteins but lacked the pre-S/S gene promoters. These data demonstrate substantial and HBV genotype-dependent diversity of spliced genomes from which a variety of aberrant precore/core fusion proteins and normal X protein but no functional envelope and P proteins could be expressed. These genomes and the encoded proteins may play a role in the viral life cycle, persistence, and pathogenesis.

Cloning, Molecular↗

Naturally occurring hepatitis B virus genomes bearing the hallmarks of retroviral G-->A hypermutation.

Two hypermutated genomes of hepatitis B virus (HBV) were cloned from sera of chronic virus carriers. Twelve percent and 26% of guanosine residues were replaced by adenosine, with the transitions being erratically distributed along the genome. G-->A substitutions showed a strong dinucleotide preference, decreasing in the order GpA > GpG > > GpC > or = GpT. Such traits are typical of retroviral G-->A hypermutation which results from cDNA synthesis coinciding with fluctuations in the intracellular [dTTP]/[dCTP] ratio. The observations offer an explanation for the high prevalence of HBV variants bearing a tryptophan 28-->stop codon in the pre-core region of carriers with chronic active or fulminant hepatitis. The HBV hypermutants indicate that a small proportion of hepatocytes have distorted dNTP pools, which might have implications for the fidelity of hepatocyte DNA replication or repair.

Adenine↗

Ultrasonic characterization of tissues via backscatter frequency dependence.

Phantom and patient studies were performed to assess the potential of backscatter frequency dependence as a useful parameter for tissue characterization. A commercial phased-array ultrasonic scanner was adapted to allow digitization of the intermediate-frequency ultrasonic data. Studies of agar phantoms containing polystyrene microspheres with 3.5 and 5 MHz transducers indicated the ability for robust differentiation of phantoms having different scatterer size and frequency dependence, based on calculated differences in mean frequencies of backscattered spectra. Using a 3.5-MHz probe, significantly lower mean frequency of ultrasound backscattered from cirrhotic, compared to normal, liver tissue was noted. Studies of benign and malignant liver tumors (hemangiomas and metastases, respectively) indicated differences in frequency content of these tumors, compared to the adjacent normal liver, that were statistically significant for 3.5-MHz data.

Adult↗

Female-specific gene expression in Schistosoma mansoni is regulated by pairing.

Gene expression studies in adult females of Schistosoma mansoni cultured in vitro revealed that the transcription of female-specifically expressed genes is influenced by pairing. In contrast, the activity of genes that are expressed in both genders was not affected by contact with the male. The transcription of genes was monitored in paired, separated and remated females. The transcript level of female-specifically expressed genes decreases within a few days following separation from males. Remating of uncoupled females with males leads to the reinitiation of transcription. These results provide strong evidence for the influence of the male on gene transcription in the female and contribute a molecular basis for the classical histological observation that the maturation of females is male dependent. The data also show that the culture system is suitable to monitor gene expression and, furthermore, they indicate de novo RNA synthesis in vitro.

Animals↗

Dynamic cell structures for the evaluation of keypoints in facial images.

In this contribution Dynamic Cell Structures (DCS network) are applied to classify local image structures at particular facial landmarks. The facial landmarks such as the corners of the eyes or intersections of the iris with the eyelid are computed in advance by a combined model and data driven sequential search strategy. To reduce the detection error after the processing of the sequential search strategy, the computed image positions are verified applying a DCS network. The DCS network is trained by supervised learning with feature vectors which encode spatially arranged edge and structural information at the keypoint position considered. The model driven localization as well as the data driven verification are based on steerable filters, which build a representation comparable with one provided by a receptive field in the human visual system. We apply a DCS based classifier because of its ability to grasp the topological structure of complex input spaces and because it has proved successful in a number of other classification tasks. In our experiments the average error resulting from false positive classifications is less than 1%.

Algorithms↗

An RNA stem-loop structure involved in the packaging of bovine leukemia virus genomic RNA in vivo.

An RNA secondary structure of the bovine leukemia virus (BLV) 5'-terminal RNA sequence was constructed by computer-assisted RNA secondary structure analysis. Mutations were created in the noncoding region (NCR) of BLV, which contains a conserved consensus sequence, to disrupt predicted secondary structure of this region. After transfection of these constructs into FLK-BLV cells and analysis of viral particles a reduction in mutant RNA content was observed relative to that of unmutated vector RNA. The packaging efficiency of the mutant with a substitution in the consensus sequence was reduced threefold and that of the mutant with a deleted 5' NCR was reduced fivefold. We conclude that predicted RNA secondary structure and/or nucleotide sequence of the BLV noncoding region is essential for BLV RNA packaging in vivo.

Animals↗

Normal renal blood flow measurement using phase-contrast cine magnetic resonance imaging.

RATIONALE AND OBJECTIVES: This study assesses the ability of a cardiac-gated phase-contrast magnetic resonance imaging (MRI) technique to measure renal blood flow (RBF) noninvasively in humans. METHODS: In nine normal volunteers, total RBF in the renal arteries and in the left renal vein was estimated by MRI and correlated with RBF determined by the clearance of para-aminohippuric acid (CPAH) and the hematocrit level. RESULTS: Correlation of RBF estimated from left renal vein flow, with RBF by CPAH-hematocrit, yielded r = .86 (P less than .003). Repeated measurement of RBF by MRI demonstrated a high degree of reproducibility, with coefficients of variation ranging from 4.8% to 8.9%. However, the MRI measurements of arterial flow did not significantly correlate with the standard measurements. CONCLUSIONS: Reproducible noninvasive measurement of normal RBF is possible with the phase-contrast MRI technique used to measure renal venous blood flow.

Adult↗

In vitro proton spectroscopy of normal and abnormal prostate.

Previous biochemical and 13C NMR spectroscopic data have suggested that the metabolism of citrate, a secretory product of normal prostate, may be interrupted in prostate cancer. In the present study in vitro 1H NMR spectroscopy was used to see if cell strains derived from prostate cancers could be reliably distinguished from those of normal prostate epithelium. High-resolution one-dimensional and two-dimensional J-resolved 1H NMR spectra as well as gas chromatography coupled with mass spectroscopy were used to study extracts of highly defined cell strains from normal peripheral zone, normal central zone, adenocarcinoma, and benign prostatic hyperplasia. Resonances assigned to citric acid and related metabolites were identified. Cell strains derived from prostate cancers tended to have smaller amounts of citrate than those from normal prostate epithelium. However, the differences were small and not statistically significant. The lack of statistically significant differences may reflect the variability present in both normal and abnormal cell strains and thus underscore the well-known difficulty in differentiating normal and cancerous tissues.

Adenocarcinoma↗

Differential therapy with calcium antagonists in pulmonary hypertension secondary to COPD. Hemodynamic effects of nifedipine, diltiazem, and verapamil.

In 53 patients with COPD and precapillary pulmonary hypertension, we investigated the effect of three typical calcium antagonists on hemodynamics at rest and during bicycle ergometer exercise. In the responders, the decrease in pulmonary vascular resistance following nifedipine was 23 percent at rest (p less than 0.0005) and 35 percent during exercise (p less than 0.0005); following diltiazem, it was 10 percent at rest (p less than 0.05) and 23 percent during exercise (p less than 0.025); following verapamil, it was 22 percent at rest (p less than 0.005) and 11 percent during exercise (p less than 0.025). The cardiac index rose significantly at rest and under exercise only after the administration of nifedipine (+16 percent and +8 percent, resp). Nifedipine caused the most distinctive peripheral vasodilation. The heart rate increased slightly following nifedipine and decreased slightly following diltiazem and verapamil. After long-term therapy with nifedipine (13 +/- 5 months), the decrease in pulmonary artery pressure and pulmonary vascular resistance was no longer significant. In our opinion, the different hemodynamic action profiles will have consequences for the differential therapy in patients with COPD and pulmonary hypertension.

Calcium Channel Blockers↗

A species and organ specific glomerular basement membrane antigen.

By the use of a monoclonal antibody reacting with the human glomerular basement membrane exclusively we have been isolating a specific glomerular antigen. The antibody failed to react with other renal structures and other basement membranes and extracellular matrix constituents of other organs or plasma proteins. It did not react with glomerular basement membranes of other species as mice and rats. For the isolation of the antigen we applied affinity chromatography, on Sepharose beads coupled to the monoclonal antibody PM II 34 G3. From this column the antigen was eluted under acid conditions. In 8% polyacrylamide gel electrophoresis the approximate molecular weight was estimated with 14400 daltons, which was confirmed on high pressure liquid chromatography using the Bio-Sil TSK method. The antigen was temperature sensitive in that at high temperatures (60 degrees C) several bands on SDS-PAGE and several peaks on HPLC could be noted. This could be responsible for the crystal formation after treatment/concentration on the rotary vacuum pump at 60 degrees C. Preliminary data of amino acid analysis showed a high glycine content pointing towards a collageneous molecule. But cross reactivity with many connective tissue proteins could be ruled out by immuno-histochemical techniques and affinity chromatography. A major point of interest is that this antigen can be detected in human urine under physiological conditions. We are herewith reporting the first species and organ specific glomerular basement membrane antigen.

Amino Acids↗

Portal venous blood flow in cystic fibrosis: assessment by Duplex Doppler sonography.

Changes of portal venous hemodynamics were investigated in 32 patients with cystic fibrosis (CF) with a mean age of 11.6 years. Hepatic profile of these patients included total bilirubin and albumin together with determination of size and echogenicity of the liver and spleen, determination of the diameter of the portal vein by real-time sonography and quantitation of flow volume of the portal vein using Duplex Doppler sonography. As a control, 35 age matched healthy children were also examined. Diameter of the portal vein was significantly increased in CF patients versus controls. Comparison of the mean flow volume of the portal vein showed a significant increase in CF-patients over 12 y old versus controls. In patients less than 12 y no significant difference of flow volume of the portal vein between CF-patients and controls was noted. The increase of diameter and flow volume of the portal vein suggest an adaptive mechanism in the pressure-volume relationship of the portal venous system in patients with CF.

Adolescent↗

pH-controlled continuous cultivation of mycoplasmas.

The continuous cultivation of mycoplasmas in a pH-controlled metabolistat was investigated with the fermentative strain Mycoplasma mobile 163K and the nonfermentative strain Mycoplasma arthritidis ISR1. The addition of medium and the removal of culture suspension were regulated by acid production from glucose by M. mobile 163K and by ammonium production from arginine by M. arthritidis ISR1, respectively. For both strains the optimal pH for continuous growth was 7.0. The steady state could be maintained for at least 21 days. With CFU of 8.4 X 10(9) ml-1 (M. mobile 163K) and 3.2 X 10(9) ml-1 (M. arthritidis ISR1), the cell concentrations were slightly higher than those obtained in batch cultures. The dependence on the adjusted pH values was measured for several parameters, such as flow rate, CFU, glucose fermentation or production of ammonia, and gliding velocity. Since the long lag phases of batch cultures can be avoided, pH-controlled continuous cultures provide an appropriate system for the production of mycoplasma cells.

Acetates↗

Hemodynamic significance of the paraumbilical vein in portal hypertension: assessment with duplex US.

Hemodynamic evaluation of portal and umbilical venous flow with duplex ultrasound (US) was performed in 11 patients with cirrhosis of the liver and a large umbilical vein. Two of these patients had hepatofugal flow in the umbilical vein exceeding hepatopetal flow in the portal vein. These two patients had no evidence of esophageal varices and bleeding. The remaining nine patients had esophageal varices. In these patients, the hepatopetal flow in the portal vein exceeded the hepatofugal flow through the umbilical vein. The authors conclude that duplex US may help identify the massive hepatofugal flow through a large umbilical vein that may reduce the likelihood of esophageal varices and variceal bleeding.

Adult↗

Prognostic value of supine scintigraphic heart function and simultaneous conventional exercise response in patients after myocardial infarction.

A total of 240 survivors of one or more myocardial infarctions were consecutively admitted to perform supine exercise radionuclide ventriculography. Within 3 years after the test, 22 died; this group was compared to an age-matched control group of 22 survivors for left and right ventricular function during rest, exercise, and simultaneously assessed exercise performance as well as ECG variables. Evaluation of 3-year survival by linear discriminant analysis revealed an accuracy of 82% for discriminant models using ECG and exercise performance variables. Implementation of resting left ventricular ejection fraction and change of right ventricular ejection fraction during exercise, as well as scintigraphic presence or absence of dyskinesia, improved the accuracy of the model to 91% of correctly classified patients.

Adult↗

Radionuclide detection of mild valvular regurgitation: its significance as assessed by Doppler sonography.

Radionuclide ventriculography (RNV) indices of regurgitation, Fourier amplitude ratio (FAR) and additional RNV variables were prospectively compared to Doppler echocardiography (DE) in 108 consecutive patients with no or mild left ventricular regurgitation, to assess RNV accuracy in detecting regurgitation in patients with different cardiac disorders. Exclusion of left ventricular or tricuspid regurgitation allowed investigation of the FAR range at rest and during exercise in a sufficiently large appropriate reference group without regurgitation. FAR, as well as other RNV variables, failed to provide more information for the diagnosis of mild (clinically irrelevant) left ventricular regurgitation than the diagnosis upon admission alone. Despite the superiority of DE as a gold standard in the detection of mild regurgitation, at present evaluation of RNV regurgitation indices might be the only method to discover regurgitation arising during dynamic exercise.

Echocardiography↗