Search PubMed⌕ Search

Biomedical subjects

E Wolf

Publications and source records attributed to E Wolf.

At least 145 records · Page 8Linked to original sources
PubMed1999

[Xenotransplantation. Possibilities of animal breeding].

The pig is the most likely donor organism for xenotransplantation of organs to humans. However, since this constellation is discordant, hyperacute rejection needs to be overcome. This review summarises current strategies of genetically modifying pigs for xenotransplantation. Limitations of the classical method of DNA-microinjection and new perspectives arising from the possibility of cloning animals from cultured cells are discussed.

Animals↗
PubMed1999

Changing incidence of mycobacterial diseases in German patients with HIV infection.

Before the introduction of potent antiretroviral therapy, disease caused by atypical mycobacteria was a frequent diagnosis in patients with fewer than 100 CD4+ cells/microL and could affect virtually every organ. The diagnosis was associated with a survival of usually less than 1 year, and antimycobacterial treatment could extend the life span by only several months.. However, the clinical face of the HIV epidemic has changed profoundly since 1996, most probably due ot the antiretroviral combination strategies. In this discussion, we assess the influence of the introduction of antiretroviral combination therapies on the incidence of mycobacterial infections from 1994 to 1998 in a cohort of German HIV-seropositive patients treated at an HIV outpatient clinic.

AIDS-Related Opportunistic Infections↗
PubMed1998

Affected enzyme activities in Alzheimer's disease are sensitive to antemortem hypoxia.

Many enzyme activities in Alzheimer's disease (AD) are changed. Some of these enzyme activities are related to certain neurotransmitter systems. Enzymes in the brain can also be sensitive to antemortem hypoxia. In the present study it was determined if enzyme activities that are altered in AD are also subject to alteration by antemortem hypoxia. As an indicator of antemortem hypoxia brain lactate concentration was used. Enzyme activities measured were those of prolyl endopeptidase (PE), aminopeptidase (AP), phosphatidylinositol (PI) kinase, phosphatidylinositol phosphate kinase, alpha-ketoglutarate dehydrogenase (alpha-KGDH), choline acetyltransferase and beta-glucuronidase. All of these enzyme activities have been measured in AD patients before and several of them have been found to be decreased. In accordance with previous findings, PE, alpha-KGDH and ChAT activities were reduced in AD patients. PI kinase and beta-glucuronidase activities, however, were not reduced, contrary to previous findings. All enzyme activities, except that of beta-glucuronidase, correlated with brain lactate concentration, suggesting that antemortem hypoxia has a major influence on the activity of enzymes in the brain. PE, AP, alpha-KGDH and ChAT activities were still different between AD and control samples when these were matched for lactate concentration. The enzyme activities that were changed in AD were also significantly correlated with lactate concentration, an indicator of antemortem hypoxia, in brain specimens. This suggests that antemortem hypoxia and AD have some factor in common that may be responsible for changes in enzyme activities. Since both PE and alpha-KGDH are known to be sensitive to oxidative stress this factor could be oxidative stress.

Adult↗
PubMed1998

Nuclear transfer in mammals: recent developments and future perspectives.

A clone can be defined as a set of genetically identical animals. Small clones of two or occasionally up to four identical animals can be obtained by embryo splitting or blastomere separation. Embryo cloning by nuclear transfer involves the transfer of genetic material from a donor cell (karyoplast) to the cytoplasm of an oocyte or zygote from which the genetic material has been removed (cytoplast). In farm animals, metaphase II oocytes are most widely used as cytoplasts. There are now many factors known to influence the efficiency of embryo cloning by nuclear transfer. These include stage of development and cell cycle of donor cells, the choice of the recipient cell, the methods for activation of oocytes, the cell cycle coordination between donor cell and recipient cytoplast, and the method for fusion between nuclear donor and recipient cytoplast. Recent progress in cloning embryos and animals from cultured cells of embryonic, fetal, or adult origin offers a wide spectrum of potential applications of nuclear transfer, such as the unlimited multiplication of elite embryos or animals from selected matings and the potential for precise genetic modification of farm animals for gene farming or xenotransplantation.

Animals↗
PubMed1998

Effect of chronic GH overproduction on cardiac ANP expression and circulating ANP levels.

Sodium and water retention are common in acromegaly and upon GH administration. The underlying mechanisms, however, have not been clearly characterized as yet. Therefore, the aim of this study was to examine possible alterations of atrial natriuretic peptide (ANP), an endogenous regulator of volume homeostasis, in response to chronic elevated GH. We used GH-transgenic mice (GH-TM) as a model for chronic hypersomatotropinemia and moreover investigated 7 and 27 week old animals, respectively, in order to discriminate between short and long term effects of GH overexpression. Hematocrit values were reduced in GH-TM compared to control animals and it is known that plasma volume is increased in these animals. Structural lesions of the kidney were found in the GH-TM, however, in the animals studied there were no signs of renal insufficiency as evidenced by serum creatinine and urea levels. The serum concentration of immunoreactive ANP (IR-ANP) determined by RIA was significantly (P < 0.005) elevated in the young GH-TM as compared to control littermates (81.7+/-13.3 vs. 50.9+/-10.8 fmol/ml). The increase in serum IR-ANP of 27 week old GH-TM, however did not reach the level of significance (57.13+/-16.3 vs. 50.25+/-16.4 fmol/ml). Serum samples of control mice as well as of the 7 week old animals mainly contained ANP 99-126, known to be the circulating form of ANP. In contrast, serum of 27 week old GH-TM predominantly showed the cardiac storage form of ANP, ANP 1-126. Cardiac expression of ANP was quantified by Northern blot analysis. mRNA coding for ANP was found 1.2- and 2-fold increased in the atria of 7 and 27 week old GH-TM, respectively. In parallel, a 2.2-fold (7 week) and 2-fold (27 week) increase of IR-ANP was observed in transgenic atria compared to tissue of control animals. In contrast, no significant difference of ANP mRNA expression or of content of IR-ANP was observed in the ventricles of both groups of animals. In conclusion, GH-TM show various alterations in their ANP status suggesting an influence of the peptide on the effect of GH in fluid retention.

Aging↗
PubMed1998

Non-linear summation of excitatory synaptic inputs to small neurones: a case study in spinal motoneurones of the young Xenopus tadpole.

1. We examined the steady-state summation of postsynaptic potentials (PSPs) in small, electrotonically compact neurones with short dendrites, using a one-compartment electrical equivalent model of the passive membrane with conductances to represent chemical synapses and electrotonic junctional connections to neighbouring neurones. 2. Our model shows that PSP summation is non-linear and for small depolarizations is mainly determined by the increase in total neurone conductance due to the opening of synaptic channels. At bigger depolarizations the change in synaptic driving force becomes an equally important cause of non-linearity. 3. Non-linear summation of AMPA-mediated PSPs was measured experimentally when two monosynaptic pathways to motoneurones were stimulated. The conductances underlying these PSPs were calculated relative to the resting neurone conductance using our model. These conductance ratios were hardly affected by the size of electrotonic coupling conductances. The non-linearity in PSP summation could be predicted by the model provided that the depolarizations remained negative to potentials at which voltage-dependent channels open. 4. The model was used to estimate the relative contributions of glutamatergic, cholinergic and electrotonic excitation to EPSPs measured in Xenopus tadpole spinal motoneurones during swimming. Estimates of synaptic conductances and electrotonic coupling to other motoneurones suggest that ligand-gated conductance mediated by glutamate may be twice that due to acetylcholine. 5. We conclude that in small electrotonically compact motoneurones of the Xenopus tadpole, our simple model can predict the non-linearity in PSP summation and may allow the conductances of different synaptic inputs to be compared. Furthermore, excitatory synaptic conductances can increase the resting neurone conductance significantly and limit depolarization. Our general model may also be applicable to other small neurones.

Animals↗
PubMed1998

Longitudinal distribution of components of excitatory synaptic input to motoneurones during swimming in young Xenopus tadpoles: experiments with antagonists.

1. Recent studies have revealed that the excitatory synaptic input to spinal motoneurones during fictive swimming in Xenopus tadpoles has three main components: glutamatergic (Glu) from premotor excitatory interneurones, nicotinic cholinergic (nACh) from more rostral motoneurones, and electrotonic coupling from neighbouring motoneurones. During swimming, these components sum to produce two kinds of excitation: phasic excitation (EPSPs) underlying spikes, and tonic depolarization. 2. We have investigated the longitudinal distribution of these excitatory synaptic inputs to presumed motoneurones at different positions along the spinal cord using intracellular recording techniques. Different antagonists (10 microM dihydro-beta-erythroidine (DHbetaE) for nicotinic ACh receptors (nAChRs), 2 mM kynurenate (Kyn) for glutamate receptors (GluRs), and 100 microM Cd2+ for all chemical synapses) were microperfused very locally to unmask the relative contributions of these components to the total excitatory drive, and their distribution along the spinal cord during swimming. 3. If the potentials remaining when all chemical components were blocked by Cd2+ were subtracted from potentials recorded after blocking nAChRs and GluRs with DHbetaE plus Kyn, a small unidentified component was observed. This component was blocked by the specific AMPA antagonist 6-nitro-7-sulphamoylbenzo(f)quinoxaline-2,3-dione (NBQX, 5 microM), so is glutamate mediated. 4. We used the potential measurements to calculate the relative synaptic conductances of the different synaptic inputs, and conclude that: (a) there is a rostral-caudal gradient in input during EPSPs and tonic depolarization; (b) the glutamatergic component accounts for most of the excitation, and decreases caudally; (c) cholinergic and electrotonic components are relatively constant in different positions along the spinal cord; and (d) these two components provide an increasing proportion of the input in more caudal neurones. 5. We propose that the glutamate components of excitation are fundamental to rhythm generation in the brainstem and rostral cord, while the electrotonic and cholinergic components ensure that the central pattern generator activates motoneurones effectively in all parts of the spinal cord.

Animals↗
PubMed1998

Insulin-like growth factor-binding protein-2 inhibits proliferation of human embryonic kidney fibroblasts and of IGF-responsive colon carcinoma cell lines.

So far, the physiological role of insulin-like growth factor binding protein-2 (IGFBP-2) has not been demonstrated directly. Therefore, we transfected 293 cells with an expression vector containing the CMV promoter and the complete cDNA of mouse IGFBP-2. Secretion of bioactive IGFBP-2 into conditioned medium was demonstrated by Western ligand and Western immunoblotting and quantified by specific RIA. For the analysis of cell proliferation three clones exhibiting either high or low/no IGFBP-2 expression were selected and compared to non-transfected parental 293 cells. IGFBP-2 secreting clones displayed reduced conversion of thiazolyl blue when compared to negative clones or non-transfected parental 293 cells (P < 0.01). The lower growth activity measured in the IGFBP-2 secreting clones was compensated in great part by the administration of exogenous IGF-I or -II. Conditioned media of IGFBP-2 secreting clones inhibited growth of IGF-responsive colon tumor cell lines (LS513, HT-29) while those of negative clones did not. In addition, conditioned medium from a clone expressing high levels of IGFBP-2 inhibited anchorage-independent growth of LS513 and HT-29 cells. In contrast, growth of an IGF-unresponsive tumor cell line (Co-115) was not affected by the conditioned media. We hypothesize that IGFBP-2 might sequester the IGFs and thus prevent them from transferring their mitogenic signals.

Animals↗
PubMed1998

Crystal structure of a GCN5-related N-acetyltransferase: Serratia marcescens aminoglycoside 3-N-acetyltransferase.

The X-ray structure of a canonical GCN5-related N-acetyltransferase (GNAT), Serratia marcescens aminoglycoside 3-N-acetyltransferase, bound to coenzyme A (CoA) has been determined at 2.3 A resolution. The single domain alpha/beta protein resembles a cupped right hand wrapped around a cylinder and consists of a highly curved, six-stranded beta sheet of mixed polarity that is sandwiched between four alpha helices. The structure includes all four conserved GNAT motifs (C, D, A, and B) and represents the catalytic core of this large enzyme superfamily. Acetyl CoA recognition is mediated by a betaalpha structure derived from GNAT motif A, which presents an invariant Arg/Gln-X-X-Gly-X-Gly/Ala segment for hydrogen bonding with the cofactor. Motif B contributes acidic residues to the binding site for the positively charged antibiotic substrate.

Acetyltransferases↗
PubMed1998

Comparison of ultrasound-guided vs laparoscopic transvaginal ovum pick-up (OPU) in simmental heifers.

Transvaginal ovum pick-up (OPU) offers several advantages over standard embryo transfer procedures. For a systematic comparison of the ultrasound-guided (U-OPU) and the laparoscopic OPU (L-OPU) method, groups of Simmental heifers were subjected to 1 of the 2 OPU-methods for 8 wk (15 sessions) followed by a treatment-free interval of 11 wk and then another 8-wk period of OPU using the alternative method. Parameters that were evaluated included the number of follicles aspirated, number and morphology of recovered cumulus-oocyte complexes (COCs), and developmental capacity of oocytes after in vitro maturation (IVM) and fertilization (IVF). Blood samples were also taken from the donors twice a wk for progesterone measurement. To evaluate effects of long-term OPU on subsequent fertility of donors, all heifers (n = 14) were inseminated during the first natural estrus after the OPU experiment. The proportion of Class I oocytes was significantly (P < 0.001) higher after U-OPU than after L-OPU (38.7% vs 21.0%). Following IVM/IVF, this difference in oocyte quality was reflected by the cleavage rate (U-OPU: 58.1%; L-OPU: 52.1%; P < 0.05) and the rate of development to morulae and blastocysts (U-OPU: 27.1%; L-OPU: 13.9%; P < 0.001). Among other factors, the greater changes in vacuum pressure during L-OPU vs U-OPU might be responsible for the difference in oocyte quality. This problem may be overcome by a more flexible system for regulating the vacuum. Progesterone levels were higher during the L-OPU than the U-OPU periods. Seven donors (50%) were diagnosed pregnant by ultrasonography on Day 28 and by palpation per rectum on Day 42.

Animals↗
PubMed1998

Composition of parental mitochondrial DNA in cloned bovine embryos.

We have investigated parental mitochondrial DNA (mtDNA) in cloned bovine embryos obtained by intraspecific cytoplast-blastomere fusion. Analysis of two-cell to blastocyst stage embryos revealed that in contrast to the exclusion of paternal (sperm) mtDNA during sexual inheritance in the cytoplast-blastomere fusion complexes, there was mixing and co-existence of parental mtDNA. The mixing of mtDNA was non-balanced with the minority deriving from the blastomere. The constant content of mtDNA during embryogenesis until the blastocyst stage suggesting an absence of mtDNA replication was shown for conventional 'in vitro fertilised' (IVF) embryos and for cloned embryos. The ratio of parental mtDNA was in accordance with the estimated quantitative participation of mtDNA from the fusion partners.

Animals↗
PubMed1998

Non-balanced mix of mitochondrial DNA in cloned cattle produced by cytoplast-blastomere fusion.

We have investigated the transmission of parental mitochondrial DNA (mtDNA) in three clones of born cattle obtained by intraspecific cytoplast-blastomere fusion. Using allele-specific TaqMan PCR a low level transmission of blastomere mtDNA (DB mtDNA) into the cloned offspring was detected, thereby generating a heteroplasmic population of mtDNA. The amount of DB mtDNA was 13% and 18% in two animals of a clone which derived from a 24-cell morula and 0.6% and 0.4% in two calves of clonal origin derived from a 92-cell morula. These values are in accordance with the tendency expected for neutral mtDNA segregation that the fewer cell divisions that have occurred in the donor embryo, the higher the amount of DB mtDNA. We also found a strong decrease of DB mtDNA which was about three orders of magnitude in the third clone derived from a 52-cell morula stage.

Animals↗
PubMed1998

A reliable procedure for differential staining of in vitro produced bovine blastocysts: comparison of tissue culture medium 199 and Ménézo's B2 medium.

A reliable double-dye technique has been established for counting the number of inner cell mass and trophectoderm cells of in vitro produced bovine blastocysts. The latter were first incubated in a 1:2 dilution of a rabbit antiserum raised against a mixture of recombinant bovine interferon tau and serum containing medium conditioned by in vitro produced trophoblastic vesicles for 45 min at 39 degrees C. Subsequently, the blastocysts were incubated in a 5% (v/v) solution of guinea pig complement in phosphate-buffered saline containing 50 micrograms/ml propidium iodide for 45 min at 39 degrees C. Then the blastocysts were transferred to ice-cold absolute ethanol containing 25 micrograms/ml bisbenzimide and evaluated under a fluorescence microscope. Since trophectoderm cells were permeabilised by antibody-mediated complement lysis, they were stained by propidium iodide (red or pink). Bisbenzimide can enter lysed and non-lysed cells and therefore stained also inner cell mass cells (blue) which had been protected from complement lysis by trophectoderm cells. This modified procedure proved to be very reliable for differential cell staining of bovine blastocysts produced under various culture conditions. A comparison of blastocysts produced in Ménézo's B2 vs. TCM 199 media (both supplemented with 10% serum from cows at oestrus) revealed significant (P < 0.01) differences in total cell numbers (119 +/- 24 vs. 84 +/- 10; mean +/- SD) and in the numbers of trophectoderm cells (79 +/- 19 vs. 57 +/- 8) and inner cells mass cells (40 +/- 7 vs. 26 +/- 5) between the two culture systems. The modified staining procedure presented here is a valuable tool for evaluating the quality of in vitro produced bovine blastocysts and for improving of culture conditions.

Animals↗
PubMed1998

Asymmetries in sensory pathways from skin to motoneurons on each side of the body determine the direction of an avoidance response in hatchling Xenopus tadpoles.

1. When swimming is initiated by tail stimulation in hatchling Xenopus tadpoles, the first trunk contraction is usually on the opposite side and directs the animal away from the stimulus. We have investigated how asymmetries in the skin sensory pathways mediate this response. 2. In alpha-bungarotoxin-immobilized tadpoles, intracellular recordings were made of responses to ipsilateral (ISS) and contralateral skin stimulation (CSS) in thirty-two presumed motoneurons. ISS evokes an inhibitory postsynaptic potential (IPSP) followed by an excitatory postsynaptic potential (EPSP) whereas CSS only evokes an EPSP. Blocking the short latency IPSP evoked by ISS with strychnine reduced the difference in spike latency on the two sides but spikes still occurred first to CSS. 3. Motoneuron EPSPs evoked by ISS and CSS were therefore recorded during microperfusion of strychnine to block the short latency IPSP. We found: (a) the CSS-EPSPs have lower threshold, larger amplitude at a given intensity of stimulus, faster rising phase, and shorter latencies than those of ISS-EPSPs; (b) the ISS-EPSP onset latencies were longer than CSS-EPSPs and became shorter as the stimulus intensity increased while those of CSS-EPSPs remained little changed. At high stimulus intensities, EPSPs caused by CSS and ISS became similar; and (c) onset latencies of ISS-EPSPs had higher variance than those of CSS-EPSPs. However, this difference was reduced as the stimulus intensity was increased. 4. Since motoneuron EPSP onset latencies varied with stimulus intensity, we proposed that the pathway from the opposite side had stronger synapses from afferents to sensory interneurons. To test this proposal we built a neuronal population model of the spinal pathway from skin afferents, via sensory interneurons to ipsilateral and contralateral motoneurons incorporating this asymmetry. Inhibition was omitted from the model. 5. Simulated motoneuron EPSPs in response to skin stimulation on each side of the body showed the major asymmetries found experimentally. If the distribution and axonal projections of the interneurons in the two sensory pathways were made the same these differences remained. However, if the synaptic strength from sensory afferents onto interneurons projecting to the two sides were made equal, the difference between the two sides were lost. 6. We propose that the sensory pathway to contralateral motoneurons has more effective excitation from afferents to sensory interneurons which leads to these motoneurons firing first. At higher stimulus strengths, when population recruitment can blur these subtle differences in excitation between the two sides, inhibition normally plays a significant role to ensure that most first responses are still contralateral.

Action Potentials↗
PubMed1998

Intestinal dimensions of mice divergently selected for body weight.

BACKGROUND: Selection for body weight provides important animal models for studying mechanisms of growth regulation. This study evaluated growth responses of the gastrointestinal tract (GIT) to long-term selection of mice for high (H line) or low (L line) 8-week body weight as compared with random-bred controls (C line). METHODS: Weights and dimensions of the various parts of the GIT were recorded from 8-month-old mice. For light microscopic stereological analyses, samples were taken from eight equidistant locations covering the whole jejunum/ileum. Vertical sections were used for estimation of fractional volumes of mucosa, submucosa, and muscularis and of villous surface area density and for measurement of villus length. RESULTS: Differences between groups in weights and dimensions of the various parts of the GIT were more pronounced in the proximal than in the distal segments, with greatest values in H, followed by C and L mice. Relative to body weight, intestinal growth was similar in the three lines, except for significantly (P < 0.001) increased relative weights of jejunum/ ileum, caecum, and colon in L mice. The fractional volume of mucosa and villus length decreased, whereas the fractional volumes of submucosa and muscularis increased from the proximal to the distal locations. The absolute volume of mucosa was greatest in H mice, followed by C and L mice. Relative to body weight, the volume of mucosa was significantly (P < 0.01) greater in L mice than in the two other lines. The mean total villous surface area of jejunum/ileum was significantly (P < 0.01) different among the three lines (L line: 144 cm2; C line: 227 cm2; H line: 304 cm2) but proportionate to differences in metabolic body weight. CONCLUSIONS: Selection for body weight affected various parts of the GIT to a different extent. The parameters investigated changed markedly along the small intestine, demonstrating the need for systematic sampling. Vertical section stereology provides unbiased estimates of total villous surface area, which is a parameter of major biological significance.

Animals↗
PubMed1998

Topography of growth hormone receptor expression in the bovine embryo.

Using in situ hybridization, mRNA encoding the growth hormone receptor (GHR) was localized in preimplantation embryos produced by in vitro fertilization (IVF) as well as in 30- to 70-day-old bovine embryos. In IVF embryos the transcript of GHR was demonstrated in the inner cell mass of 8-day-old blastocysts. In 30-day-old embryos, the mesonephros was the first organ to express the mRNA of GHR. In 40-day-old embryos, the transcript was found in the neurones of the spinal ganglions, the splanchnic nerves and the motoneurones of the spinal cord, in the vascular endothelium, and in the developing striated muscle tissue. Colocalization of the protein by immunohistochemistry showed an identical distribution pattern of GHR in 30- to 70-day-old embryos.

Animals↗
PubMed1998

Altered growth of mice divergently selected for body weight is associated with complex changes in the growth hormone/insulin-like growth factor system.

Mice investigated in this study were generated by selecting a sub-population of the NMRI out-bred stock (C), for high (H) or low (L) 8-week body weight. After 58 generations of selection, 8-week body weights of the sub-populations were markedly different if compared to controls. To investigate endocrine changes causing the altered growth performance in the different lines of mice, we analysed several components of the growth hormone (GH)/insulin-like growth factor (IGF) system. Pituitary weights of male and female L mice were significantly smaller than those of C and H mice. Relative to body weight, pituitary weights of male mice from the three lines did not differ, however pituitary weight-to-body weight-ratios of female L mice were significantly greater than those of H females. Mean volume densities of somatotropic cells were significantly smaller in L mice than in C and H mice. Serum IGF-I concentrations were significantly lower in the L line than in the C and H lines. H mice displayed significantly increased serum insulin levels both after ad libitum feeding and after a 14 hour fasting period. Ligand blot analysis of serum IGF-binding proteins (IGFBPs) revealed a significant reduction of circulating IGFBP-3 in L mice as compared to C and H mice. In contrast, serum IGFBP-2 mRNA levels were significantly increased in male L mice and showed non significant increases in female L mice. Hepatic IGFBP-2 mRNA levels were significantly increased in L mice and decreased in H mice as compared to C mice. Expression of IGFBP-4 mRNA in the liver was significantly decreased in both selection lines (L, H) as compared to the random-bred controls. Our findings demonstrate that altered growth of mice resulting from selection for body weight is associated with complex changes in the endocrine network of the GH/IGF system.

Adipose Tissue↗