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R Ehrlich

Publications and source records attributed to R Ehrlich.

At least 109 records · Page 6Linked to original sources
PubMed1998

Modelling a 3D structure for EgDf1 from Echinococcus granulosus: putative epitopes, phosphorylation motifs and ligand.

EgDf1 is a developmentally regulated protein from the parasite Echinococcus granulosus related to a family of hydrophobic ligand binding proteins. This protein could play a crucial role during the parasite life cycle development since this organism is unable to synthetize most of their own lipids de novo. Furthermore, it has been shown that two related protein from other parasitic platyhelminths (Fh15 from Fasciola hepatica and Sm14 from Schistosoma mansoni) are able to confer protective inmunity against experimental infection in animal models. A three-dimensional structure would help establishing structure/function relationships on a knowledge based manner. 3D structures for EgDf1 protein were modelled by using myelin P2 (mP2) and intestine fatty acid binding protein (I-FABP) as templates. Molecular dynamics techniques were used to validate the models. Template mP2 yielded the best 3D structure for EgDf1. Palmitic and oleic acids were docked inside EgDf1. The present theoretical results suggest definite location in the secondary structure of the epitopic regions, consensus phosphorylation motifs and oleic acid as a good ligand candidate to EgDf1. This protein might well be involved in the process of supplying hydrophobic metabolites for membrane biosynthesis and for signaling pathways.

Amino Acid Sequence↗
PubMed1998

Expression of allogeneic MHC class I antigens by transgenic mouse trophoblast does not interfere with the normal course of pregnancy.

Mammalian embryos express paternal histocompatibility antigens which make them potential targets for maternal immune responses. Yet, the histoincompatible fetus survives and develops normally. Down regulation of classical MHC antigen expression by trophoblast cells which are in direct contact with maternal circulation has been repeatedly shown. The trophoblast cells are unable to function properly in antigen presentation and do not induce allogeneic rejection reactions. In the present study we have created transgenic mice that express an allogeneic class I transgene whose transcription is controlled by the transferrin receptor promoter. The expression patterns of the transgene product mice from a single transgenic line were studied in each of the typical placental subpopulations. The allogeneic class I antigen was expressed in the allantoic plate region of the trophoblast, and this expression was not restricted to the endothelial region but extended also to the spongiotrophoblast, as well as the major blood vessels and in the endodermal sinuses. In contrast to the normal class I expression, prominent levels of allogeneic H-2 antigens were detected in the labyrinthine trophoblast. The fetal resorption rate in females mated with these transgenic males was not higher then the normal rate, and the embryos survived and developed normally. These data imply that the unusual expression of allogeneic class I antigens in certain trophoblast subpopulations does not affect fetal development.

Animals↗
PubMed1998

Lead absorption and renal dysfunction in a South African battery factory.

OBJECTIVES: To test the association between inorganic lead (Pb) exposure, blood pressure, and renal function in South African battery factory workers, with both conventional and newer measures of renal function and integrity. METHODS: Renal function measures included serum creatinine, urea, and urate (n = 382). Urinary markers (n = 199) included urinary N-acetyl-beta-D-glucosaminidase (NAG), retinol binding protein, intestinal alkaline phosphatase, tissue non-specific alkaline phosphatase, Tamm-Horsfall glycoprotein, epidermal growth factor, and microalbuminuria. RESULTS: Mean current blood Pb was 53.5 micrograms/dl (range 23 to 110), median zinc protoporphyrin 10.9 micrograms/g haemoglobin (range 1.9 to 104), and mean exposure duration 11.6 years (range 0.5 to 44.5). Mean historical blood Pb, available on 246 workers, was 57.3 micrograms/dl (range 14 to 96.3). After adjustment for age, weight and height, positive exposure response relations were found between current blood Pb, historical blood Pb, zinc protoporphyrin (ZPP), and serum creatinine and urate. Blood pressure was not associated with Pb exposure. Among the urinary markers, only NAG showed a positive association with current and historical blood Pb. CONCLUSION: An exposure-response relation between Pb and renal dysfunction across the range from < 40 to > 70 micrograms/dl blood Pb was found in this workforce, with conventional measures of short and long term Pb exposure and of renal function. This could not be explained by an effect on blood pressure, which was not associated with Pb exposure. The findings probably reflect a higher cumulative renal burden of Pb absorption in this workforce in comparison with those in recent negative studies. The results also confirm the need for strategies to reduce Pb exposure among industrial workers in South Africa.

Acetylglucosaminidase↗
PubMed1997

Differential resistance to proteinase K digestion of the yeast prion-like (Ure2p) protein synthesized in vitro in wheat germ extract and rabbit reticulocyte lysate cell-free translation systems.

The Ure2p yeast prion-like protein was translated in vitro in the presence of labeled [35S]methionine in either rabbit reticulocyte lysate (RRL) or wheat germ extract (WGE) cell-free systems. When subjected to proteinase K digestion, the Ure2p protein synthesized in WGE was proteolysed much more slowly compared to that synthesized in RRL; this displays fragments of about 31-34 kDa, persisting over 8 min. Thus, the digestion rate and pattern of the protein synthesized in WGE, unlike that synthesized in RRL, revealed characteristic features of the [URE3] prion-like isoform of the Ure2p protein [Masison, D.C. and Wickner, R.B. (1995) Science 270, 93-95]. Chloramphenicol acetyltransferase, synthesized under the same conditions, differed fundamentally in its proteolytic sensitivity toward proteinase K (PK); in the RRL system it was more slowly digested than in WGE, proving specific PK inhibitors to be absent in both systems. Posttranslational addition of the WGE to the RRL-synthesized Ure2p does not protect Ure2p from efficient PK degradation either. The differences in Ure2p degradation may be ascribed to a specific structure or specific states of association of Ure2p synthesized in WGE; obviously, they yield a protein that mimics the behavior of the Ure2p in [URE3] yeast strains. The present data suggest that particular conditions of the Ure2p protein translation and/or certain cellular components (accessory proteins and extrinsic factors), as well as the nature of the translation process itself, could affect the intracellular folding pathway of Ure2p leading to the de novo formation of the prion [URE3] isoform.

Animals↗
PubMed1997

Adenovirus E1a interferes with expression of vaccinia viral genes.

The 12S and 13S cDNAs of the oncogene E1a encoded by the early region of adenovirus 12 (Ad12) were overexpressed using the T7/encephalomyocarditis (EMC)/vaccinia hybrid expression system. The E1a proteins were stable for at least 12 h in monkey epithelial BSC1 cells. The E1a proteins were recognized by a rabbit polyclonal antibody and displayed phosphorylation patterns similar to those displayed by the E1a proteins expressed in Ad12-transformed cells. Expression of E1a proteins by recombinant vaccinia virus led to inhibition of vaccinia viral protein synthesis which was observed as soon as 6 h after infection. This suppression was mediated by both the 12S and the 13S products of Ad12E1a and to a somewhat lesser extent by the 13S product of Ad2E1a. The inhibition of vaccinia virus gene expression resulted in enhanced survival of vaccinia virus-infected cells. These results suggest that the proteins encoded by the E1a sequester a viral or a cellular product(s) that is essential for the expression of vaccinia virus-encoded genes.

Adenoviridae↗
PubMed1997

Synthesis and turnover of beta2-microglobulin in Ad12-transformed cells defective in assembly and transport of class I major histocompatibility complex molecules.

In primary embryonal fibroblasts from transgenic mice expressing H-2 genes and a miniature swine class I transgene (PD1), transformation with the highly oncogenic Ad12 results in a reduction in peptide transporter and proteasome-associated (LMP2 and LMP7) gene expression, and suppression in transport and cell surface expression of all class I antigens. The selective suppression in transport of H-2 (but not of PD1) molecules in cells reconstituted for the expression of peptide transporter and LMP genes implied that an additional factor(s) is involved in the assembly of class I complexes. Here we show that the beta2m, H-2Db, and H-2Kb genes are transcribed and translated in Ad12-transformed cells. However, unlike normal and E1Ad5-transformed cells, in which beta2m is either secreted unbound or bound to class I heavy chains, in Ad12-transformed cells significant amounts of beta2m are retained in the cell bound to the membrane, but free of class I heavy chains. This abnormal turnover of beta2m in the Ad12-transformed cells suggests the existence of a novel beta2m-binding molecule(s) that sequesters beta2m, and this process may provide a mechanism by which transformation with Ad12 may subvert class I complex formation.

Adenoviruses, Human↗
PubMed1997

Remarks on the phylogeny and structure of fatty acid binding proteins from parasitic platyhelminths.

Four fatty acid binding proteins (FABPs) have been described in 4 parasitic platyhelminths: Schistosoma mansoni, Schistosoma japonicum, Fasciola hepatica and Echinococcus granulosus. FABPs form a multigenic family of cytosolic proteins widely distributed in metazoan tissues, the function of which is still poorly understood. These helminth proteins have recently received attention, since there are reports to indicate that S. mansoni and F. hepatica FABPs may be protective antigens. In addition, these proteins could play a major role in the parasites' life-cycles because platyhelminths are unable to synthesize de novo most of their lipids. We have undertaken phylogenetic and structural analyses of platyhelminth FABPs in an attempt to characterize features of biological relevance. Phylogenetically, these FABPs appear to be more closely related to those of vertebrate heart, mammary gland, muscle, retina, skin, brain and myelin, although no clear functional relationships were established between them. We describe several conserved motifs characteristic of specific groups of FABPs. Hydrophilicity, flexibility and accessibility analyses revealed several major putative epitopes for the E. granulosus FABP, EgDf1, that appear to be centred in loops of the EgDf1 3-dimensional structure modelled by molecular replacement.

Amino Acid Sequence↗
PubMed1997

The homeobox-containing gene EgHbx3 from Echinococcus granulosus is expressed in the stalk of protoscoleces.

The homeobox-containing gene EgHbx3 from Echinococcus granulosus is expressed in the stalk of protoscoleces. The expression of the homeobox-containing gene EgHbx3 from E. granulosus was studied in protoscoleces by in situ hybridisation techniques. A method for performing in situ hybridisations on whole mounted hydatic material is described. Our results show a clear positive signal at the protoscolex stalk, when using the antisense EgHbx3 probe. This signal is particularly strong in the proximal end of the stalk (with respect to the protoscolex).

Animals↗
PubMed1997

Modulation of antigen processing and presentation by persistent virus infections and in tumors.

Cell-mediated immunity is effective against cells harboring active virus replication and is critical for the elimination of ongoing infections, opposing tumor progression, and reducing or preventing the reactivation of persistent viruses and tumor metastasis. The capacity of persistent viruses and tumor cells to maintain a long-term relationship with their host presupposes mechanisms for circumventing antiviral or antitumor defenses. By suppressing the expression of molecules associated with antigen processing and presentation, abrogation of the major immune mechanism that deals with the elimination of infected and transformed cells is achieved. This is accomplished in tumors predominantly by transcriptional downregulation of genes encoding class I major histocompatibility complex antigens, peptide transporter molecules, and the proteasome-associated low molecular mass protease subunits, and in cells expressing viral proteins by interfering with peptide transport and the assembly/transport of class I complexes. In addition, virus-infected cells and selected tumor cells express mainly nonimmunogenic or antagonistic peptide epitopes. This review describes mechanisms used by viruses and in transformed cells for interference with antigen processing and presentation and addresses their significance for in vivo viral persistence and tumor progression.

Adenoviridae↗
PubMed1997

Presence of a conserved domain of GATA transcription factors in Echinococcus granulosus.

In order to characterize GATA transcription factors in Echinococcus granulosus, a PCR-based cloning strategy was developed. Degenerate oligonucleotides were designed for the most conserved sequence in GATA proteins that include 20 amino acids of the zinc domain. A 60 bp fragment was isolated that had high homology among this sequence and those reported in other species. An analogous sequence was obtained by performing the same procedure with DNA from the free living platyhelminth Dugesia tigrina. High stringency Southern blotting experiments confirmed the presence of this sequence in the parasite genome.

Amino Acid Sequence↗
PubMed1997

Expression and immunogenicity of an Echinococcus granulosus fatty acid-binding protein in live attenuated Salmonella vaccine strains.

Fatty acid-binding proteins (FABPs) are candidate molecules for vaccines against several parasitic platyhelminths. A FABP from the cestode Echinococcus granulosus (EgDf1) was expressed in Salmonella vaccine strains as a C-terminal fusion to fragment C of tetanus toxin (TetC) by using expression vector pTECH. The fusion protein was equally expressed in several attenuated vaccine strains derived from bacteria with different genetic backgrounds and different attenuating mutations. Single-dose immunization experiments with the aroA Salmonella typhimurium strain SL3261 carrying the pTECH-EgDf1 construct were conducted with mice, using both the intravenous and the oral routes. Surprisingly, the antibody response to EgDf1 and the antigen-specific cytokine production in spleen cells were stronger in mice immunized orally. Furthermore, immune mouse sera strongly reacted with fixed sections of the worm's larval stage. Analysis of the isotype distribution of the specific anti-EgDf1 antibodies showed similar production of immunoglobulin G1 (IgG1) and IgG2a together with specific IgA antibodies. In addition, stimulation of spleen cells from mice immunized with the different constructs with either Salmonella lysate, TetC, or EgDf1 showed that, together with Th1-related cytokines (gamma interferon and interleukin 2 [IL-2]), significant levels of a Th2 cytokine (IL-5) were produced specifically, indicating a Th2 component to the response to the Salmonella carrier and to the recombinant antigens. Salmonellae expressing the TetC-rEgDfl fusion are currently under evaluation as potential vaccines against E. granulosus.

Animals↗
PubMed1997

Prevalence of occupational lung disease among Botswana men formerly employed in the South African mining industry.

OBJECTIVE: To determine whether previous health experiences affect the prevalence of occupational lung disease in a semirural Botswanan community where there is a long history of labour recruitment to South African mines. METHOD: A cross sectional prevalence study of 304 former miners examined according to a protocol including a questionnaire, chest radiograph, spirometry, and medical examination. RESULTS: Overall mean age was 56.7 (range 28-93) years, mean duration of service 15.5 (range 2-42) years. 26.6% had a history of tuberculosis. 23.3% had experienced a disabling occupational injury. Overall prevalence of pnemoconiosis (> 1/0 profusion, by the International Labour Organisation classification) was 26.6%-31.0%, and 6.8% had progressive massive fibrosis (PMF). Many were entitled to compensation under South African law. Both radiograph readers detected time response relations between pneumoconiosis and PMF among the 234 underground gold miners. PMF could result from < 5 years of exposure, but was not found < 15 years after first exposure. Both pulmonary tuberculosis (PTB) and pneumoconiosis were found to be associated with airflow limitation. CONCLUSIONS: Former miners in Botswana have a high prevalence of previously unrecognised pneumoconiosis, indicative of high previous exposures to fibrogenic respirable dust. Their pneumoconiosis went unrecognised because they had no access to surveillance after employment. Inadequate radiographic surveillance or failure to act on results when employed or when leaving employment at the mines could have contributed to under recognition. Community based studies of former miners are essential to fully evaluate the effects of mining exposures. Our findings indicate a failure of established measures to prevent or identify pneumoconiosis while these miners were in employment and show that few of the social costs of occupational lung diseases are borne by mining companies through the compensation system.

Accidents, Occupational↗
PubMed1997

Keeping an eye on Crohn's disease: orbital myositis as the presenting symptom.

Episodic periorbital swelling due to presumed orbital inflammation and myositis caused intermittent apparent proptosis and was the presenting symptom of ileocecal Crohn's disease (CD) in a teenage female with a family history of autoimmune disorders and CD. Orbital myositis, a very rare extraintestinal manifestation of inflammatory bowel disease (IBD), likely represents a process of impaired immunoregulation related to the underlying intestinal inflammation. This rare manifestation of IBD simulates the more commonly encountered thyroid orbitopathy (ophthalmopathy), but IBD should be considered if all thyroid tests are negative. It is important to recognize that orbital myositis may be an extraintestinal manifestation of Crohn's disease so that the diagnosis can be made and appropriate therapy commenced.

Autoimmune Diseases↗
PubMed1996

Characterization of a flatworm ribosomal RNA-encoding gene: promoter sequence and small subunit rRNA secondary structure.

The transcription start point (tsp) of a ribosomal RNA (rRNA)-encoding gene (rDNA) from Echinococcus granulosus has been mapped at a position located 1.1 kb upstream from the small subunit (SSU) rRNA coding sequence. As expected from the analysis of the putative promoter sequence (-200 to +50), no homology was found with rDNA promoters from other organisms. Nevertheless, some interesting motifs were found. There is a d(T)11 track flanked by an inverted repeat (IR) centered at position -32, which is analogous to the position of the TATA box in promoters transcribed by RNA polymerase II. Two other IR were observed that are also present in the Trypanosoma cruzi rDNA promoter. We have also determined the SSU rDNA sequence and established a secondary structure model. The analysis of the secondary structure strongly suggests that this gene encodes a functional SSU rRNA. The fact that both the promoter and the rRNA coding sequence are derived from the same rDNA repeat indicates that the promoter is also functional.

Animals↗