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Biomedical subjects

M Engels

Publications and source records attributed to M Engels.

At least 19 recordsLinked to original sources

Bioflocculent algal-bacterial biomass improves low-cost wastewater treatment.

An innovative technology for the biological treatment of wastewater in regions with sufficient solar radiation based on the simultaneous growth and degradation processes of algal and bacterial biomass is presented. The aim of the work is the improvement of pond technology through the formation of stable algae-bacteria aggregates, which a) permit a simple separation of the algal biomass by gravity sedimentation, b) enable a high removal efficiency for organic carbon and nutrients, and c) are independent in terms of oxygen provision through algal photosynthesis. Algae-bacteria aggregates could be developed with a suitable algal species (Chlorella vulgaris, Strain Hamburg) as a 'model organism' in a wastewater environment. The morphology of algal-bacterial flocs is similar to activated sludge flocs. They are stable and settle quickly. Floc size ranged between 400 and 800 microm. Results of our experiments with an artificially irradiated lab-scale system, operated in continuous flow mode, revealed that even at a relatively short hydraulic detention time of two days, a high elimination capacity of 9.96 g N m(-2) d(-1) and 0.87g Pm(-2) d(-1) can be achieved. Recent investigations confirmed that floc formation of unicellular algae and wastewater bacteria also could be developed and maintained in a pilot-scale system with a water depth of 0.5 m.

Bacteria↗

[Porcine malignant catarrhal fever: diagnostic findings and first detection of the pathogenic agent in diseased swine in Switzerland].

For the first time Ovine Herpesvirus 2 (OvHV-2) was identified in Swiss pigs as the causative agent of Porcine Malignant Catarrhal Fever (MCF). Diseased animals from two farms were observed to show weakness, anorexia, fever up to 41 degrees C, and neurological symptoms, i.e. ataxia, convulsions and hyperesthesia, erosion on the snout and in the oral and nasal mucosa, as well as multiple skin lesions. Histopathological findings included severe non-purulent inflammation with mononuclear cell infiltration in several organs. Most dominant were meningo-encephalitis, disseminated nephritis as well as purulent catarrhalic bronchopneumonia. The findings were quite reminiscent of the lesions due to MCF in cattle and give therefore substantial proof to use Porcine Malignant Catarrhal Fever as the term for the disease. Identification of the causative agent was done with a quantitative PCR specific for OvHV-2. Different tissues from diseased animals were positive. Furthermore, one animal which had been ill for more than five days tested positive for antibodies against an epitope conserved among MCF viruses. Serum samples from diseased animals reacted negative towards Classical Swine Fever- and Pseudorabies virus antigen. A weakly positive reaction against porcine enterovirus type I argued against the involvement of enteroviruses in the observed disease. Moreover, by means of different conventional PCRs, we detected the newly discovered porcine lymphotropic herpesviruses for the first time in Switzerland and could at the same time exclude their involvement in Porcine Malignant Catarrhal Fever.

Animals↗

Transduction of Vero cells and bovine monocytes with a herpes simplex virus-1 based amplicon carrying the gene for the bovine herpesvirus-1 Circ protein.

Herpes simplex virus-1 (HSV-1) based amplicon vectors are promising gene delivery vehicles because they have a large transgene capacity and can efficiently transduce many different cell types, including non-dividing cells, of various animal species. The Circ protein of bovine herpesvirus-1 (BHV-1) is a myristylated virion component of unknown function. Preliminary experiments with a circ gene deletion mutant indicated that Circ may influence the host's immune response by downregulating MHC-II expression in bovine monocytes. To get more insight into the function of Circ, amplicon vectors were constructed with various open reading frames (ORFs) under the control of the HSV-1 IE4/5 promoter: (i) the Circ ORF alone, (ii) a fusion ORF encoding an N-terminal Circ fused to the enhanced green fluorescent protein (eGFP), (iii) the eGFP ORF alone, and (iv) the Circ ORF in the inverted orientation. Upon helpervirus-free packaging into HSV-1 amplicon particles and transduction of Vero cells, both Circ alone and the Circ-eGFP fusion protein produced a punctate pattern within the cytoplasm, suggesting membrane association of the myristylated protein. In contrast, eGFP alone was evenly distributed over the cytoplasm of transduced cells. Upon infection of bovine buffy-coat cells, it was observed that cells of the monocyte lineage but not lymphocytes were transduced. Transgene expression reached a peak around 20h after transduction and lasted for at least 90h. Transduced monocytes underwent specific morphological changes, which may be attributed to Circ synthesis.

Animals↗

Latency and reactivation of bovine herpesvirus 1 (BHV-1) in goats and of caprine herpesvirus 1 (CapHV-1) in calves.

Bovine Herpesvirus 1 (BHV-1) and Caprine Herpesvirus 1 (CapHV-1) are related members of the herpesvirus family. Since their natural hosts are often kept in close contact with each other, concern was raised that a reservoir might be established in the heterologous host in addition to the homologous host. To investigate this possibility, cross-infection experiments with BHV-1 in goats and CapHV-1 in calves were performed. BHV-1 infected goats developed mild disease signs during acute infection, whereas CapHV-1 infection in calves took a subclinical course. However, virus excretion and antibody production were indicative of successful cross-infection of both BHV-1 and CapHV-1. Reactivation of BHV-1 was achieved in 5 out of 8 goats as demonstrated by recurrent virus excretion and rising antibody titers. In constrast CapHV-1 in calves could not be reactivated experimentally. Nevertheless, PCR revealed that both viruses established latency in the trigeminal ganglia of the heterologous host. Based on these results we conclude that goats should indeed be regarded as a potential BHV-1 reservoir, which must be considered during IBR eradication programs.

Animals↗

Biomass, productivity and density of the seagrass Thalassia testudinum at three sites in Cahuita National Park, Costa Rica.

The basic ecology of seagrass beds was investigated by comparing biomass, productivity and density of Thalassia testudinum (turtle grass) at three sites: Puerto Vargas, Punta Cahuita and Rio Perezoso, in Cahuita National Park, Limón, Costa Rica, over a two month period (March-April 1999). Above ground biomass, density, and productivity were highest in the Puerto Vargas site while Punta Cahuita had the least non-green above ground biomass was significantly lower in total biomass than Puerto Vargas. Punta Cahuita was distinguished by the largest grain size, a very hard substrate, and shallower water. Rio Perezoso, on the other hand, had extremely fine sediment and lower salinity, while Puerto Vargas was intermediate both in sediment size and environmental conditions. It appears, therefore, that higher biomass and productivity result from a combination of moderate environmental characteristics and an intermediate sediment size.

Biomass↗

Restriction endonuclease analysis of the genome of two Italian caprine herpesvirus 1 strains.

Two caprine herpesvirus (CpHV.1) strains isolated in two goat flocks in southern Italy were compared with E/CH and McK/US reference CpHV.1 strains by restriction endonuclease analysis. BamHI, KpnI, NotI and PstI restriction enzymes were used. With these enzymes McK/US strain was clearly differentiated from the others. KpnI and NotI digestion resulted in unique patterns for all four strains. Restriction enzyme analysis might be a useful method to distinguish CpHV.1 isolated from animals and provide a basis for further epidemiological studies of the CpHV.1 infection.

Alphaherpesvirinae↗

Glycoprotein C of bovine herpesvirus 5 (BHV-5) confers a distinct heparin-binding phenotype to BHV-1.

Bovine herpesvirus type 1 (BHV-1) causes respiratory and genital diseases in cattle, whereas the closely related BHV-5 can induce severe meningoencephalitis in calves. To characterize BHV-5 glycoprotein C (gC5) within the backbone of BHV-1, three consecutive recombinant viruses were constructed: A deletion mutant (rBHV-1delta gC blue) with gC replaced by the lacZ gene, an exchange mutant (rBHV-1gC5) with the lacZ of BHV-1delta gC blue exchanged by gC5, and a rescue mutant (rescue BHV-1) from rBHV-1gC5 with an additional XbaI site in gC1. The recombinant and wildtype viruses were characterized on MDBK cells. Although no significant differences were observed in growth behaviour and entry kinetics, rBHV-1gC5 showed a distinct phenotype in a heparin blocking assay. The gC5 was able to transfer the heparin binding phenotype of BHV-5 to BHV-1. This indicates that gC1 and gC5 differ in their receptor binding qualities, which might modulate the ability of the viruses to spread within the central nervous system.

Alphaherpesvirinae↗

Novel entry pathway of bovine herpesvirus 1 and 5.

Herpesviruses enter cells by a yet poorly understood mechanism. We visualized the crucial steps of the entry pathway of bovine herpesvirus 1 (BHV-1) and BHV-5 by transmission and scanning electron microscopy, employing cryotechniques that include time monitoring, ultrarapid freezing, and freeze substitution of cultured cells inoculated with virus. A key step in the entry pathway of both BHV-1 and BHV-5 is a unique fusion of the outer phospholipid layer of the viral envelope with the inner layer of the plasma membrane and vice versa resulting in "crossing" of the fused membranes and in partial insertion of the viral envelope into the plasma membrane. The fusion area is proposed to function as an axis for driving the virus particle into an invagination that is concomitantly formed close to the fusion site. The virus particle enters the cytoplasm through the opened tip of the invagination, and the viral envelope defuses from the plasma membrane. There is strong evidence that the intact virus particle is then transported to the nuclear region.

Animals↗

Radiation-induced impairment of urinary bladder function. Assessment of micturition volumes.

PURPOSE: The present investigation was initiated in order to study the feasibility of repeat micturition volume measurements for the determination of acute variations in urinary bladder function in radiotherapy patients. PATIENTS AND METHODS: Thirty-seven consecutive patients with malignancies of the prostate (11), rectum (9), corpus uteri (11), or cervix uteri (6) were included in this pilot study. All patients were treated according to standard irradiation protocols. All patients were asked to present with maximum bladder filling for each radiation treatment. After irradiation, patients were weighed before and after micturition, and the weight difference was assumed to reflect the micturition volume. RESULTS: No systematical variations in bladder capacity could be assessed during treatment for uterine tumors. In rectum carcinoma patients, a decrease in micturition volume by about 20% was observed between weeks 2 and 5 which, however, was not statistically significant; subsequently, normal to supra-normal values were measured. In patients treated for prostate cancer, volumes were reduced from week 2; in weeks 5 to 6, minimum values of approximately 70% of the control volumes were seen, which represent a significant reduction (p < 0.05). CONCLUSIONS: Functional changes represent the majority of side effects of radiation therapy in the urinary bladder. The assessment of urinary bladder function by pre- and post-micturition weighing is a feasible and sensitive method to objectively determine radiation-induced changes. This method may be applied for the documentation of acute radiotherapy side effects.

Documentation↗

Pathogenesis of ruminant herpesvirus infections.

Ruminants are hosts for members of both Alpha- and Gamma-herpesvirinae. A wide range of disease syndromes is associated with infections by these agents. The associated diseases reflect the biological nature of the causative viruses. Clinically, the symptoms may be mild and localized or include severe generalized disease, leading eventually to death. Much knowledge has been gained concerning the pathogenesis of some alpha-herpesviruses. Initially, these viruses replicate in epithelial cells at the portal of entry. The symptoms of the acute diseases are often associated with the destruction of those epithelial cells. However, as in the case of bovine herpesvirus 1 (BHV-1), the virus may spread in the infected host by viremia, gaining access to a broader range of tissues and organs, and causing a broader variety of diseases. Furthermore, many herpesviruses are capable of entering neuronal cells. There, they may replicate, which may lead to neuronal diseases, for example, encephalitis. In addition, the herpesviruses may establish latency in neuronal or lymphoid cells. During latency, apparently no viral antigens are synthesized but the genomes of the latent viruses are present in the nuclei of long living cells, such as, e.g., neurones of the ganglia corresponding to the sites of peripheral replication. Upon reactivation, the viruses re-establish the lytic cycle of replication. Shielded from the effectors of the immune system, they migrate back to the peripheral tissues where they are excreted and may be transmitted. Although a strong immune response is provoked during primary viral replication, these mechanisms help the herpesviruses to escape from immune surveillance during latency and to a lesser degree during reactivation. It has been observed that certain herpesviruses may behave differently upon infection of different hosts. Relatively little progress has been made concerning the understanding of the pathogenesis of ruminant herpesviruses but much has been learned about viral molecular biology. Many viral proteins have been identified and characterized and the technology to create recombinant viruses has been established. With these tools in our hands, it is now possible to address the really interesting questions concerning pathogenesis. We postulate that herpesviruses contain at least two sets of genes, a first set involved in gene expression and viral replication, and a second set responsible for functions, which may affect pathogenesis, latency, and virus/host interactions. Using recombinant virus technology, it will be possible in the future to design targeted deletions and gene transfers in ruminant herpesviruses in order to study the viral and host factors involved in pathogenesis on the molecular level.

Alphaherpesvirinae↗

Targeted molecular dynamics: a new approach for searching pathways of conformational transitions.

Molecular dynamics simulations have proven to be a valuable tool to investigate the dynamic behavior of stable macromolecules at finite temperatures. However, considerable conformational transitions take place during a simulation only accidentally or at exceptionally high temperatures far from the range of experimental conditions. Targeted molecular dynamics (TMD) is a method to induce a conformational change to a known target structure at ordinary temperature by applying a time-dependent, purely geometrical constraint. The transition is enforced independently of the height of energy barriers, while the dynamics of the molecule is only minimally influenced by the constraint. Simulations of decaalanine and insulin show the ability of the method to explore the configurational space for pathways accessible at a given temperature. The transitions studied at insulin comprise unfolding of an alpha-helical portion and, in the reverse direction, refolding from an extended conformation. A possible application of TMD is the search for energy barriers and stable intermediates from rather local changes up to protein denaturation.

Computer Graphics↗

Identification and zinc dependence of the bovine herpesvirus 1 transactivator protein BICP0.

Bovine herpesvirus 1 (BHV-1) specifies and unspliced early 2.6-kb RNA (ER2.6) which is 3' coterminal with exon 2 of the 2.9-kb immediate-early (IE) RNA. The two transcripts have a common open reading frame (676 codons). The predicted protein, designated BHV-1 infected cell protein 0 (BICP0), contains a zinc finger domain with homology to ICP0 of herpes simplex virus type 1 and protein 61 of varicella-zoster virus, and depending on the promoter, it acts as a strong activator or as a repressor in transient expression assays. In situ immunoadsorbent assays using antisera against synthetic oligopeptides demonstrated that BICP0 accumulates in nuclei of BHV-1-infected cells, as expected for an IE gene product involved in gene regulation. Western blots (immunoblots) revealed a BHV-1-specific 97-kDa protein which was detectable during the IE phase and also at later periods of infection, indicating that the kinetics of BICP0 synthesis is consistent with the switch from IER2.9 to ER2.6. To confirm that ER2.6 encoded the 97-kDa BICP0 protein, a DNA fragment containing BICP0-coding sequences was inserted into the Autographa californica baculovirus genome. A recombinant protein, identified by its reactivity with antipeptide sera, exhibited the same electrophoretic mobility as BICP0 specified by BHV-1. We microinjected Xenopus oocytes with a BICP0 effector plasmid and a promoter-chloramphenicol acetyltransferase plasmid. BICP0-induced stimulation of this promoter was strongly reduced when intracellular zinc was chelated by thionein, indicating that the effect of BICP0 is zinc dependent.

Amino Acid Sequence↗

Modeling of G-protein coupled receptors with bacteriorhodopsin as a template. A novel approach based on interaction energy differences.

The structure of bacteriorhodopsin was used as a template to generate a model for G-protein coupled receptors. However, these receptors and the template are not related by sequence homology. Therefore a pragmatic and reproducible approach was developed to achieve an energetically favourable accommodation of receptor sequences to the backbone structure of bacteriorhodopsin. Improved interaction energy differences are used in a two step procedure analogous to a hypothetical folding mechanism for integral membrane proteins. The resulting model is in good agreement with existing data from structure-function studies.

Amino Acid Sequence↗

Recent evaluation of prognostic risk factors in esophageal atresia--a multicenter review of 223 cases.

In this study, 223 cases of esophageal atresia (Type IIIb: 85.7%; Type II: 5.8%; Type IIIc: 4.0%; Type IIIa: 2.2%; Type IV: 2.2%) from 6 pediatric surgery centers of Austria, were retrospectively examined for the following parameters and their influence on the prognosis: Birth weight (2494.7 +/- 702.0 g), gestation week (range 27-42 weeks; mean 37.3 +/- 3.1 weeks), sex (male: n = 128; female: n = 95), long-gap atresia (> or = 2 cm: n = 33), Tracheomalacia (n = 16), associated malformations (n = 122; cardiac 27.4%, renal 17.9%, skeletal 17.0%, anal: 10.3%, intestinal 9.9%, mediastinal 7.6%, chromosomal 2.2%), preoperative aspiration (n = 92), pneumonia (n = 96), anastomotic insufficiency (n = 45), empyema (n = 5), mediastinitis (n = 8), sepsis (n = 32), other medical complications (n = 122, in 80 infants), other surgical complications (n = 57). The mortality rate was 41.3% overall, from 1975 to 1991; however, it was 25% from 1987 to 1991 and 0% in 1991. A statistically significant correlation was found between prognosis and the following factors: Cardiac malformations (p = 0.0001), medical complications except aspiration and pneumonia (p = 0.0001), empyema (p = 0.0081), mediastinitis (p = 0.0214), and sepsis (p = 0.0295). These 5 significant factors were given different points and a prognostic score was calculated by the addition of these points. This score was predictive for survival in 90.6% of cases and for mortality in 94% of cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Austria↗

Resuscitation of the very immature infant: cerebral Doppler flow velocities in the first 20 minutes of life.

Blood flow velocity of the anterior cerebral artery was investigated by Doppler ultrasonography in five time intervals up to 20 min after birth in 16 non-asphyxiated very immature infants (median birth weight 1,058 g, range 720-1,930 g; median gestational age 30 weeks, range 27-32 weeks) delivered by caesarean section and intubated after birth. Heart frequency, oscillometric mean arterial blood pressure and blood gases were recorded as well. There was a transitory increase in both systolic and end-diastolic velocities (from 29 to 35 and from 1 to 10 cm/s) during the first 5 min after birth which occurred together with an increase in heart frequency. The overall incidence of intracranial haemorrhage was low (3/16, 19%). This observation study shows a transitory increase in cerebral blood flow velocity with a peak at about 5 min after birth in preterm infants < 33 weeks of gestation undergoing standard resuscitation.

Birth Weight↗

Interactions of bovine and caprine herpesviruses with the natural and the foreign hosts.

Bovine herpesvirus 1 (BHV1) and caprine herpesvirus 1 (CapHV1) are useful models to study virus-host interactions, as well as pathogenicity and latency, when comparing the outcome of infection in the natural and the foreign hosts. Molecular seroepidemiological analyses revealed that cross-reacting antibodies were mainly induced by glycoprotein gI (gB analogue), by the major capsid protein and by nonstructural proteins, whereas the most virus-specific antibodies were elicited by glycoproteins gIII and gIV. These glycoproteins, especially gIII (gC analogue), might therefore play an important role in the virus-host-interactions. As a basis for further studies, we re-evaluated observations concerning experimental infections with BHV1 and CapHV1 in the natural and the foreign hosts. All parameters indicated that both viruses were able to infect either host, but that the pathogenicity was restricted to the natural host. Latent virus could be reactivated exclusively from cows infected with BHV1. It was possible neither to reactivate BHV1 from goats, nor to reactivate CapHV1 from either species. The experiments indicated that the outcome of infection in the natural and the foreign host is dependent on host and viral factors, whereby gIII is only one important virus component involved. Further investigations in the host and host cell range of BHV1 and CapHV1 will help to clarify the role of factors responsible for virus-host-interactions.

Animals↗

The T<-->R structural transition of insulin; pathways suggested by targeted energy minimization.

The transition of insulin between its crystallographically defined states T and R is connected with considerable change even of backbone structure: the N-terminal B chain (residues B1-B8) refolds from extended conformation in T into helical in R, and vice versa. Although hitherto observed only in hexamers the transition of the monomer was adequate for developing and testing the method of 'targeted energy minimization' (TEM), capable of coping with conformational changes of such extent at moderate computational expenditure. The simulation is performed in a predetermined number of steps consisting of two atomic displacements each, one by force in the direction of the target structure, the second by energy minimization releasing the constraint caused in the first. The transition pathway is represented by the string of energy minimized transient structures. Due to the directedness of the algorithm the simulated pathway for R-->T is not the reversal of that for T-->R. It is, therefore, not pretended that the minimum energy pathway was identified. In the T-->R direction the N-terminal B chain first swivels while remaining largely stretched and then winds up extending the pre-existing helix B9-B19. The A chain advances into the space abandoned and withdraws from it in the R-->T simulation. In the latter the extended helix first kinks at B8/B9, and then the B1-B8 segment is unwound and stretched. The helical H-bonds of that segment are formed late in T-->R and are maintained during almost half of R-->T. The AN helix is less stable and more involved in the transitions than helix AC.(ABSTRACT TRUNCATED AT 250 WORDS)

Algorithms↗

[Schoenlein-Henoch syndrome with abdominal manifestations without skin involvement].

The etiology of Schoenlein-Henoch' Syndrome has not yet been fully clarified [10, 15]. An allergically toxic genesis is under discussion [8, 19]. The classical combination of symptoms consists of urticariel efflorescences, bleeding of skin and lining tissue and arthralgies [8, 19]. Involvement of kidneys and abdomen may occur [8, 19]. Our case report concerns exclusively an abdominal form with colics, vomiting and diarrhea [10, 18, 16]. Chemical tests reveal a reduction of factor XIII [1, 5, 21]. There is no involvement of kidneys, skin or joints. With reference to literature and this particular case, etiology, diagnosis and therapy are dealt with.

Child↗