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E Petersen

Publications and source records attributed to E Petersen.

At least 73 records · Page 4Linked to original sources
PubMed1996

Molecular cloning and homology modeling of protocatechuate 3,4-dioxygenase from Pseudomonas marginata.

The genes that encode the alpha and beta subunits of protocatechuate 3,4-dioxygenase (3,4-PCD [EC 1.13.11.3]) were cloned from a Pseudomonas marginata genomic library. These genes pcaG and pcaH, were found when screening the library for hydrolase genes. The two open reading frames of the PCD genes could be identified adjacent to an esterase gene by sequence homology. A 1.7-kb KpnI/ApaI fragment, carrying pcaG and pcaH, was subcloned and the genes were functionally expressed in Escherichia coli. The deduced amino acid sequence shows high homology to previously determined amino acid sequences of bacterial protocatechuate 3,4-dioxygenases. A homology model based on the available crystal structure of the protocatechuate 3,4-dioxygenase from Pseudomonas aeruginosa shows high similarity with the binding and catalytic sites.

Amino Acid Sequence↗
PubMed1996

Recombinant Toxoplasma gondii surface antigen 1 (P30) expressed in Escherichia coli is recognized by human Toxoplasma-specific immunoglobulin M (IgM) and IgG antibodies.

The immunodominant surface antigen of Toxoplasma gondii, surface antigen 1 (SAG1), was expressed in Escherichia coli as a fusion protein containing a majority of the SAG1 protein supplied with six histidyl residues in the N-terminal end. The recombinant protein was purified on a Ni-chelate column and then on a fast-performance liquid chromatography column and was in a nonreduced condition. It was recognized by T. gondii-specific human immunoglobulin G (IgG) and IgM antibodies as well as by a mouse monoclonal antibody (S13) recognizing only nonreduced native SAG1. Antibodies induced in mice by the recombinant SAG1 recognized native SAG1 from the T. gondii RH isolate in culture. Recombinant SAG1 is suitable for use in diagnostic systems for detecting anti-SAG1-specific IgG and IgM antibodies.

Animals↗
PubMed1996

Multicenter evaluation of a fluorometric enzyme immunocapture assay to detect toxoplasma-specific immunoglobulin M in dried blood filter paper specimens from newborns.

An easy-to-perform fluorometric enzyme immunocapture assay (FEIA) was developed by Labsystems, Helsinki, Finland, to detect toxoplasma-specific immunoglobulin M (IgM) in dried blood spots. Assay materials were distributed to two sites that have programs in place designed to identify infants born with congenital toxoplasma infection: the Statens Serum Institut, Copenhagen, Denmark, and the New England Regional Newborn Screening Program, Boston, Mass. Each site tested over 700 dried blood samples from healthy newborns to define a cutoff at the 99.5 percentile (5 enzyme immunounits for Copenhagen and 4 enzyme immunounits for Boston). Each site then applied its own cutoff of interpret results for dried blood spots prepared from either adults with serology suggestive of acute infection (Copenhagen) or infants determined to be congenitally infected on the basis of serological criteria (Boston). In Copenhagen, 35 of 38 adult samples were either positive to a small degree or borderline positive for IgA. These samples thus may not represent acute infection. In Boston, of 26 congenitally infected infants, 22 were positive by FEIA. The four infant specimens not positive by FEIA were either negative or borderline positive by the standard Boston assay. These results demonstrate that the IgM FEIA is a potential alternative to other filter paper assay for toxoplasma-specific IgM currently in use for newborns.

Adult↗
PubMed1996

Comparison of two methods for enumerating malaria parasites in thick blood films.

Calculation of parasite densities is important for estimating herd immunity to malaria, and for determining end points in field trials for interventions such as malaria vaccines, impregnated bed nets, and chemosuppression. Two methods of enumeration were compared: method 1, in which 100 consecutive high-power fields (HPFs) are examined, and if they all contain at least one parasite, the number per field is then counted in 10-100 of these fields according to density; and method 2, in which the actual number of parasites present in 100 consecutive fields are counted. The first method significantly underestimates parasite density in samples in which less than all high-power fields are parasite-positive. A correction of method 1 is suggested, which results in a parasite density, which is comparable with that obtained using method 2. The correction factor estimated was 2(-In(1 - p)), where p is the proportion of positive HPFs. The correction factor presented will allow accurate estimate of parasite densities per volume of blood even if only the proportion of parasite-positive high-power fields containing at least one parasite are counted.

Animals↗
PubMed1995

[Occurrence of toxoplasmosis in pregnant women in Denmark. A study of 5.402 pregnant women].

Consecutive serum samples from 5402 women were analyzed for specific IgG and IgM Toxoplasma gondii antibodies by enzyme immunosorbent assay. The prevalence of IgG antibodies was 27.4%. The seroconversion rate was estimated to be 1.16% per year (SE = 0.0017) in all age groups, consistent with an incidence of acute toxoplasmosis in pregnant women of 0.65%. Based on theses figures, the estimated number of congenitally infected cases are discussed. There was no difference in the prevalence of antibodies in women from rural or urban areas, or from different parts of Denmark.

Acute Disease↗
PubMed1995

Toxoplasma gondii: characterization of a monoclonal antibody recognizing antigens of 36 and 38 kDa with acid phosphatase activity located in dense granules and rhoptries.

Acid phosphatase activity in Toxoplasma gondii was studied by electron microscopy and found in the rhoptries and dense granules. On Western blots, six bands were found to have acid phosphatase activity; a double band of 36-38 kDa showed the strongest activity. In order to raise monoclonal antibodies to antigens of potential importance in parasite invasion of the host cell, mice were immunized with live T. gondii of the cyst-forming ME119 isolate and later boosted with tachyzoites of the RH strain. One of the monoclonal antibodies, S16, recognized the 36- to 38-kDa antigen. Immunogold labeling and electron microscopy showed that the antigens were located in dense granules and rhoptries. Using immunoprecipitation with S16, SDS-PAGE, IEF, and Western blotting, we show that the 36- to 38-kDa rhoptry antigen recognized by S16 has acid phosphatase activity.

Acid Phosphatase↗
PubMed1995

Detection by enzyme immunosorbent assay of Toxoplasma gondii IgG antibodies in dried blood spots on PKU-filter paper from newborns.

Different ways of eluting IgG antibodies against Toxoplasma gondii from blood spots on PKU filter papers from newborns were examined, and the eluate was analysed by Enzyme Immunosorbent Assay (EIA). 10-mm diameter discs (method A), and 3-mm discs (method B) were eluted overnight, and 3-mm discs were eluted directly for 2 hours in the EIA wells (method C). Two groups of paired samples were tested: blood spots on the PKU card from the newborn, cord blood, and a venous blood sample from the mother collected shortly after delivery (Group 1, 191 samples); or PKU cards from the newborns and serum collected from the mother during the first trimester (Group 2, 426 samples). The best results were found for Group 1; method A, a sensitivity of 100% (63/63), and a specificity of 100% (128/128) (r = 0.934, P < 0.0001), and for Group 2; method C, a sensitivity of 100% (125/125), and a specificity of 100% (301/301) (r = 0.822, P < 0.0001). When method C, was used on 188 samples stored for 16-18 month at -20 degrees C, a sensitivity of 100% (62/62) and a specificity of 99.2% (125/126) (r = 0.913, P < 0.0001) were found. The PKU filter paper method is reliable and reproducible, and can be used in screening projects based on analysis of neonatal samples, and in epidemiological studies on toxoplasmosis in pregnant women.

Adult↗
PubMed1994

The significance of blood levels of IgM, IgA, IgG and IgG subclasses in Sudanese visceral leishmaniasis patients.

We developed an ELISA test using leishmania antigenic extracts to detect antigen-specific antibody responses, including subclass and isotype analysis, in visceral leishmaniasis (VL) patients from the Sudan. A total of 92 parasitologically proven patients were compared with cutaneous leishmaniasis, schistosomiasis, malaria, onchocerciasis and tuberculosis patients, as well as with healthy endemic and non-endemic controls. Some VL patients were examined before and after chemotherapy. VL patients showed significantly higher IgG responses compared with all other groups (93.4% sensitivity, 93.7% specificity), and higher (but not significantly) IgM responses. All groups showed low IgA levels. All IgG subclasses, IgG1, 2, 3, and 4, showed higher levels in patients than all other groups, with IgG1 and IgG3 levels being significantly reduced following treatment. The rank order for specificity and sensitivity for IgG subclasses was IgG3 > IgG1 > IgG2 > IgG4.

Adolescent↗
PubMed1994

Immune responses to band 3 neoantigens on Plasmodium falciparum-infected erythrocytes in subjects living in an area of intense malaria transmission are associated with low parasite density and high hematocrit value.

During the intracellular development of the human malarial parasite, Plasmodium falciparum, cryptic regions of the erythrocyte band 3 protein are exposed. Antibodies against these band 3-related neoantigens block cytoadherence, and peptides based on amino acid sequences of putative exofacial loops of band 3 protein block the in vitro and in vivo adherence of P. falciparum-infected erythrocytes. At present, it is not known whether reactivity to these antigens is related to exposure to the malaria parasite or is correlated with protective immunity. The reactivities of plasma to peptides containing amino acid sequences of putative exofacial loops 3 and 7 of human band 3 protein were determined for children and adults living in an area of perennial malaria transmission (Liberia) and for donors who had never been exposed to malaria (Denmark). Plasma samples from children and adults living in an area of intense malaria transmission showed a much higher reactivity with the band 3 peptides than did those from nonimmune individuals. High reactivity to the loop 3 peptide (amino acids 546 to 555) was correlated with lower mean parasite density in children in the 5- to 9-year-old age group. The presence of antibodies against loop 3 and 7 peptides was not associated with a low packed erythrocyte volume (hematocrit); in fact, higher-than-average reactivities to both peptides were positively correlated with high hematocrit values, indicating that antibodies which specifically recognize the band 3-related neoantigens are not involved in hemolysis (autoimmunity).

Adolescent↗
PubMed1993

Urinary excretion of modified nucleosides as biological marker of RNA turnover in patients with cancer and AIDS.

Using boronate gel affinity chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC), a method for the simultaneous determination of 12 urinary modified nucleosides has been developed. The RP-HPLC fractions were identified by gas chromatography/mass spectrometry analysis. The HPLC quantitation of urinary nucleoside levels before and after surgery of cancer patients suggested that urinary 5'-deoxy-5'-methylthioadenosine and N-[(9-beta-D-ribofuranosyl-9H-purine-6-yl) carbamoyl]-L-threonine (t6A) levels were helpful in monitoring therapeutic effects in cancer patients. From the fact that molar ratios of urinary N2,N2-dimethylguanosine (m2 2G)pseudouridine (psi) and t6A/psi in cancer patients were lower than those of normal or post-surgical cancer patients, the increase of rRNA content in cancer tissues growing rapidly was estimated using the stoichiometric relationship between the ratio of the number of residues of their modified nucleoside in RNAs and the proportion of rRNA to total RNAs in average tissues of whole body. Furthermore, from the estimation of RNA turnover using urinary nucleoside levels, it was found that the half-lives of rRNA rather than tRNA of patients with cancer and those of both RNAs in the case of acquired immunodeficiency syndrome (AIDS) were extremely short compared with those of the normal. Thus, we discovered that the selected urinary modified nucleosides were very useful as a biological marker of whole-body RNA turnover in patients with cancer and AIDS.

Acquired Immunodeficiency Syndrome↗
PubMed1993

Classification of clinical falciparum malaria and its use for the evaluation of chemosuppression in children under six years of age in Liberia, west Africa.

The possible role of malaria as cause of morbidity was assessed during one year in 262 children aged 6 months to 6 years living in two villages in a rural area of Liberia. The study population was followed by weekly clinics and three-monthly surveys and the children were randomly allocated to receive either chloroquine or placebo every 3 weeks. The morbidity of the children was evaluated by criteria based on the history and the clinical condition into four different stages, in order to describe the probability that an observed clinical event could be attributed to malaria infection, based on the presence of detectable parasites in the blood, the history the previous week, and the clinical status of the child. The level of anaemia, splenomegaly and measured body temperature supported that malaria was the major contributor to the overall morbidity observed. Based on the stage classification of clinical illness, children were classified as having 'possible clinical malaria' or 'probable clinical malaria'. Malaria appeared to be an important cause of febrile episodes during both dry and rainy seasons. During the rainy season more than 60% of the children experienced at least one clinical malaria episode, and during the dry season more than 50% of the children experienced at least one clinical attack of malaria. Children receiving chemosuppression had overall fewer clinical malaria attacks, and the effect of the chemosuppression was most pronounced in the dry season, the odds ratio comparing children receiving regular chemosuppression with children receiving presumptive treatment only was estimated to 0.39 (0.25-0.62).

Anemia↗
PubMed1993

First cases of microsporidiosis in Scandinavian patients with AIDS.

The first 2 cases of infection with Enterocytozoon bieneusi in Denmark and Scandinavia are reported. Both patients were women and to the best of our knowledge this is the first report of E. bieneusi in female AIDS patients. Both had late stage AIDS, and both had complained of intermittent diarrhoea for more than 1 year. At the time microsporidiosis was diagnosed, no other pathogens causing diarrhoea were found. Immunodeficient patients with chronic unexplained diarrhoea should be investigated for intestinal microsporidiosis, especially as treatment is now available.

AIDS-Related Opportunistic Infections↗
PubMed1993

Extra-intestinal amebiasis: clinical presentation in a non-endemic setting.

37/38 patients with reciprocal titers > or = 512 against Entamoeba histolytica in Denmark over a 5-year period were evaluated retrospectively in order to establish the clinical profile of extra-intestinal amebiasis in a non-endemic area. 24 of these had extra-intestinal amebiasis, all presenting 1 or more amebic liver abscesses on ultrasonography. Fever was the most common finding, present in 91% of the cases. The most striking observation was the lack of both abdominal pain and tenderness in 22% of the patients with liver abscess. Pulmonary symptoms and abnormal chest X-rays were each recorded in 45% of the cases. Liver abscesses with or without pulmonary involvement were the only extra-intestinal manifestations recorded. The condition was initially misconceived in more than half the cases, but most of the patients responded well once treatment with metronidazole was started.

Adult↗