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Biomedical subjects

E Fuchs

Publications and source records attributed to E Fuchs.

At least 217 records · Page 12Linked to original sources
PubMed1991

Mutant keratin expression in transgenic mice causes marked abnormalities resembling a human genetic skin disease.

To explore the relationship between keratin gene mutations and genetic disease, we made transgenic mice expressing a mutant keratin in the basal layer of their stratified squamous epithelia. These mice exhibited abnormalities in epidermal architecture and often died prematurely. Blistering occurred easily, and basal cell cytolysis was evidence at the light and electron microscopy levels. Keratin filament formation was markedly altered, with keratin aggregates in basal cells. In contrast, terminally differentiating cells made keratin filaments and formed a stratum corneum. Recovery of outer layer cells was attributed to down-regulation of mutant keratin expression and concomitant induction of differentiation-specific keratins as cells terminally differentiate, and the fact that these cells arose from basal cells developing at a time when keratin expression was relatively low. Collectively, the pathobiology and biochemistry of the transgenic mice and their cultured keratinocytes bore a resemblance to a group of genetic disorders known as epidermolysis bullosa simplex.

Animals↗
PubMed1991

[Malaria prophylaxis and self-care 1990--problems and current solutions].

For successful malaria prophylaxis, future travelers need to be informed about the risks of infection and about systematic measures to be taken against mosquito bites. For visits in areas of high transmission, continuous chemoprophylaxis is recommended, whereas in regions with low transmission a stand-by medication for self-therapy is sufficient. Each of these four measures has some problems which are discussed here in addition to the recently published recommendations by the Swiss Working Group for Medical Advice to Travelers.

Antimalarials↗
PubMed1991

Keratin genes, epidermal differentiation and animal models for the study of human skin diseases.

The examples shown here illustrate the power of gene targeting to the epidermis as a means of developing animal models for the study of human skin diseases. In this short review, I have focused on contributions which stem predominantly from my own laboratory [31, 32, 34, 37, 47]. However, other laboratories have also contributed heavily to the development of this technology [28-30, 35, 36]. The opportunities for these models are vast: there are a myriad of human skin diseases which have been characterized extensively at a biological level, but whose aetiology is presently unknown. A combined knowledge of (a) the biochemistry of epidermal differentiation; (b) epidermal-specific gene expression; and (c) transgenic mouse technology has provided the foundation for these and future studies in this area.

Animals↗
PubMed1991

Sorting out IF networks: consequences of domain swapping on IF recognition and assembly.

Vimentin and keratin are coexpressed in many cells, but they segregate into two distinct intermediate filament (IF) networks. To understand the molecular basis for the sorting out of these IF subunits, we genetically engineered cDNAs encoding hybrid IF proteins composed of part vimentin and part type I keratin. When these cDNAs were transiently expressed in cells containing vimentin, keratin, or both IFs, the hybrid IF proteins all recognized one or the other or both networks. The ability to distinguish networks was dependent upon which segments of IF proteins were present in each construct. Constructs containing sequences encoding either helix 1B or helix 2B seemed to be the most critical in conferring IF recognition. At least for type I keratins, recognition was exerted at the level of dimer formation with wild-type type II keratin, as demonstrated by anion exchange chromatography. Interestingly, despite the fact that swapping of helical domains was not as deleterious to IF structure/function as deletion of helical domains, keratin/vimentin hybrids still caused structural aberrations in one or more of the cytoplasmic IF network. Thus, sequence diversity among IF proteins seems to influence not only coiled-coil but also higher ordered associations leading to 10-nm filament formation and/or IF interactions with other cellular organelles/proteins.

Amino Acid Sequence↗
PubMed1991

A function for keratins and a common thread among different types of epidermolysis bullosa simplex diseases.

Previously we demonstrated that transgenic mice expressing a mutant keratin in the basal layer of their stratified squamous epithelia exhibited a phenotype bearing resemblance to a subclass (Dowling Meara) of a heterogeneous group of human skin disorders known as epidermolysis bullosa simplex (EBS) (Vassar, R., P. A. Coulombe, L. Degenstein, K. Albers, E. Fuchs. 1991. Cell. 64:365-380.). The extent to which subtypes of EBS diseases might be genetically related is unknown, although they all exhibit skin blistering as a consequence of basal cell cytolysis. We have now examined transgenic mice expressing a range of keratin mutants which perturb keratin filament assembly to varying degrees. We have generated phenotypes which include most subtypes of EBS, demonstrating for the first time that at least in mice, these diseases can be generated by different mutations within a single gene. A strong correlation existed between the severity of the disease and the extent to which the keratin filament network was disrupted, implicating perturbations in keratin networks as an essential component of these diseases. Some keratin mutants elicited subtle perturbations, with no signs of the tonofilament clumping typical of Dowling-Meara EBS and our previous transgenic mice. Importantly, basal cell cytolysis still occurred, thereby uncoupling cytolysis from the generation of large, insoluble cytoplasmic protein aggregates. Moreover, cell rupture occurred in a narrowly defined subnuclear zone, and seemed to involve three factors: (a) filament perturbation, (b) the columnar shape of the basal cell, and (c) physical trauma. This work provides the best evidence to date for a structural function of a cytoplasmic intermediate filament network, namely to impart mechanical integrity to the cell in the context of its tissue.

Animals↗
PubMed1991

Transforming growth factor alpha induces collagen degradation and cell migration in differentiating human epidermal raft cultures.

When cultured on plastic and treated with transforming growth factor alpha (TGF alpha), human keratinocytes exhibit an increase in proliferation at the colony periphery, apparently as a consequence of enhanced cell migration (Barrandon and Green, 1987). To investigate the effects of TGF alpha on a differentiating stratified squamous epithelium and to begin to examine the molecular basis mediating this influence, we cultured human epidermal cells on a gelled lattice of collagen and fibroblasts, floating on the air-liquid interface. Under these conditions, raft cultures differentiate and exhibit morphological and biochemical features of human skin in vivo (Asselineau et al., 1986; Kopan et al., 1987). When 3-wk-old raft cultures were treated with TGF alpha, basal cells showed a marked increase in cell proliferation. At elevated concentrations of TGF alpha, the organization of cells within the artificial tissue changed and islands of basal cells entered the collagen matrix. Biochemical analysis of the response revealed that type I collagenase and gelatinase were induced by keratinocytes within 12 h after TGF alpha treatment. In contrast, invasion of basal cells into the collagen matrix was not significant until 48-72 h post-treatment, suggesting that collagenase and gelatinase production may be a prerequisite to this phenomenon. These results have important implications for the possible role of TGF alpha in squamous cell carcinoma and tumor invasion.

Cell Differentiation↗
PubMed1991

Racial heterogeneity of HIV antigenemia in people with HIV infection.

We compared the prevalence of HIV p24 antigenemia in black and white US patients with HIV infection. The prevalence of HIV antigenemia increased with severity of HIV disease (P less than 0.001). In all clinical categories, whites were more likely to be HIV-antigenemic than blacks (overall prevalence 38 versus 18%; P less than 0.01). Anti-p24 antibodies were detected in a higher proportion of blacks (84%) than whites (65%; P = 0.02). Blacks had significantly higher total serum immunoglobulin levels than whites (median 3.8 versus 3.2 mg/dl; P less than 0.00001). Racial differences in HIV antigen expression may result from differences in humoral response to HIV infection. These differences should be considered when HIV antigen is used as a surrogate marker in clinical trials.

Adult↗
PubMed1991

Transgenic mice provide new insights into the role of TGF-alpha during epidermal development and differentiation.

Transforming growth factor-alpha (TGF-alpha) is thought to be the major autocrine factor controlling growth in epidermal cells. To explore further the role of TGF-alpha in epidermal growth and differentiation, we used a human keratin K14 promoter to target expression of rat TGF-alpha cDNA to the stratified squamous epithelia of transgenic mice. Unexpectedly, the only regions of epidermis especially responsive to TGF-alpha overexpression were those that were normally thick and where hair follicle density was typically low. This included most, if not all, body skin from 2-day- to 2-week-old mice, and ear, footpad, tail, and scrotum skin in adult mice. In these regions, excess TGF-alpha resulted in thicker epidermis and more stunted hair growth. Epidermal thickening was attributed both to cell hypertrophy and to a proportional increase in the number of basal, spinous, granular, and stratum corneum cells. During both postnatal development and epidermal differentiation, responsiveness to elevated TGF-alpha seemed to correlate with existing epidermal growth factor (EGF) receptor levels, and we saw no evidence for TGF-alpha-mediated control of EGF receptor (EGFR) expression. In adults, no squamous cell carcinomas were detected, but benign papillomas were common, developing primarily in regions of mechanical irritation or wounding. In addition, adult transgenic skin that was still both sensitive to TGF-alpha and subject to mild irritation displayed localized regions of leukocytic infiltration and granular layer loss, characteristics frequently seen in psoriasis in humans. These unusual regional and developmental effects of TGF-alpha suggest a natural role for the growth factor in (1) controlling epidermal thickness during development and differentiation, (2) involvement in papilloma formation, presumably in conjunction with TGF-beta, and (3) involvement in psoriasis, in conjunction with some as yet unidentified secondary stimulus stemming from mild mechanical irritation/bacterial infection.

Animals↗
PubMed1991

Three epidermal and one simple epithelial type II keratin genes map to human chromosome 12.

We have localized the genes which encode the human type II epidermal keratins K5, K6a, and K6b and the simple epithelial keratin K7 (KRT5, KRT6A, KRT6B, and KRT7, respectively) to chromosome 12 using Southern blot analysis of somatic cell hybrids. In addition, we have sublocalized the genes for K6a and K7 to bands 12q12----q14 on the long arm of this chromosome by in situ hybridization of metaphase chromosomes.

Blotting, Southern↗
PubMed1990

A randomized controlled trial of a reduced daily dose of zidovudine in patients with the acquired immunodeficiency syndrome. The AIDS Clinical Trials Group.

BACKGROUND: The initially tested dose of zidovudine for the treatment of patients with advanced disease caused by the human immunodeficiency virus type 1 (HIV) was 1500 mg. Although this dose is effective, it is associated with substantial toxicity. METHODS: To evaluate the efficacy and safety of a reduced dose, we conducted a randomized controlled trial in 524 subjects who had had a first episode of Pneumocystis carinii pneumonia. The subjects were assigned to receive zidovudine in either a dose of 250 mg taken orally every four hours (the standard-treatment group, n = 262) or a dose of 200 mg taken orally every four hours for four weeks and thereafter 100 mg taken every four hours (the low-dose group, n = 262). RESULTS: The median length of follow-up was 25.6 months. At 18 months the estimated survival rates were 52 percent for the standard-treatment group and 63 percent for the low-dose group (P = 0.012 by the log-rank test). At 24 months the estimated survival rates were 27 percent for the standard-treatment group and 34 percent for the low-dose group (P = 0.033). In both groups, 82 percent of the subjects had another opportunistic infection, and the length of time to that infection was similar in the two groups (P = 0.56 by the log-rank test). CD4 T-lymphocyte counts improved transiently in both groups, and serum levels of HIV antigen decreased in the subjects with antigenemia. The hemoglobin level declined to less than 5 mmol per liter (80 g per liter) in 101 subjects in the standard-treatment group and in 77 in the low-dose group (39 vs. 29 percent, P = 0.0009 by the log-rank test). The neutrophil count declined to less than 0.750 x 10(9) per liter in 134 subjects in the standard-treatment group and in 96 in the low-dose group (51 vs. 37 percent, P = 0.0001). CONCLUSIONS: The reduced daily dose of zidovudine used in this study was at least as effective as the standard dose and was less toxic; however, with the use of a four-week induction period with a high dose followed by low-dose treatment, severe anemia and neutropenia were common complications of treatment with zidovudine.

Acquired Immunodeficiency Syndrome↗
PubMed1990

Localization and quantification of [125I]-endothelin binding sites in human fetal and adult kidneys--relevance to renal ontogeny and pathophysiology.

Endothelins, 21 amino acid peptides, produced by endothelial cells are potent vasoconstrictors and mitogens. According to experimental studies in animals, endothelins seem to be involved in the regulation of renal hemodynamics. In order to gain insight into its potential effects in man, a quantitative analysis of its binding sites was performed in human kidneys. Because of the proliferative action of endothelin in cell culture we also compared binding sites in fetal and adult kidneys. Binding sites for [125I]-endothelin-1,2,3 were visualized by in-vitro autoradiography and quantified by densitometry. In both adult and fetal tissue, specific binding sites occurred in the cortex, medulla, and renal vessels. Unlabeled endothelins and sarafotoxin, a peptide with a high sequence homology to endothelins, inhibited [125I]-endothelin-1 binding with IC50 in the 9.8 to 0.023 nM range, whereas unrelated peptides (angiotensin II, atrial natriuretic peptide) and the calcium antagonist nitrendipine failed to compete for [125I]-endothelin-1 binding sites. Linear Scatchard analysis revealed that the number of binding sites (expressed per tissue equivalent: TE) were consistently higher in fetal than in adult kidneys, while affinities did not differ significantly in cortex, medulla, and vessels (fetal/adult: cortex KD 43.4 +/- 19.6/55.9 +/- 16.7 nM; BMax 13.5 +/- 7.8/2.7 +/- 1.3 fmol/mg TE; medulla KD 26.3 +/- 10.9/34.6 +/- 7.4 nM; BMax 10.1 +/- 0.9/3.7 +/- 1.1 fmol/mg TE; vessels KD 41.1 +/- 22.9/23.7 +/- 8.1 nM; BMax 12.9 +/- 3.9/4.1 +/- 1.2 fmol/mg TE). Medullary capillaries and veins showed strong binding in human and rat kidneys which may be important for the pathophysiology of acute renal failure. Human adult and fetal glomeruli had only a few binding sites. This contrasts to findings in the rat kidney in which glomeruli have a high concentration of endothelin binding sites; although this does not role out an influence per se, it does point out the need to subject the assumption of a relevant glomerular effect of endothelin in man to closer scrutiny. The diffuse and strong binding in fetal kidney may indicate a role for endothelin in the process of renal maturation.

Adult↗
PubMed1990

Alpha 2-adrenergic binding sites in the medulla oblongata of tree shrews demonstrated by in vitro autoradiography: species related differences in comparison to the rat.

Alpha 2-adrenergic binding sites in the medulla oblongata of tree shrews and rats were detected and quantified by in vitro-autoradiography with the alpha 2-antagonist 3H-rauwolscine (3H-RAUW). The autoradiographic pattern of the radioligand binding in the tree shrew medulla oblongata resembles that which has been described by others for the human myelencephalon. This pattern coincides well with the occurrence of catecholaminergic structures detected by immunocytochemistry with antibodies against phenylethanolamine-N-methyltransferase and tyrosine hydroxylase. In contrast to the rat, where only the nucleus tractus solitarii and the nucleus dorsalis nervi vagi were labeled, five discrete nuclei specifically bound 3H-RAUW in tree shrews. The highest number of binding sites was detected in the nucleus dorsalis nervi vagi (nX; Bmax: 333 fmoles/mg) and the nucleus tractus solitarii (NTS; 311 fmoles/mg), followed by the nucleus nervi hypoglossi (nXII; 297 fmoles/mg), the nucleus reticularis parvocellularis (FRS; 230 fmoles/mg), and the area of the catecholamine cell groups A1 and C1 (area C1; 202 fmoles/mg). Maximal binding in the two labeled nuclei of the rat was 158 fmoles/mg. The discrete nuclei of the two species also showed different affinities for 3H-RAUW with Kd ranging from 0.17 to 0.83 nM in tree shrews and 1.80 to 1.95 nM in rats. Competition experiments revealed that the radioligand bound specifically to alpha 2-binding sites. In the tree shrew, nX, nXII and the area C1, also have a relatively high affinity for the alpha 1-antagonist prazosin which is a quality of the adrenoceptor subtype alpha 2B. Furthermore, in the area C1, 3H-RAUW binding was inhibited by the dopamine antagonist haloperidol. There are thus species related as well as regional differences with respect to the number, the affinity, and the pharmacological properties of alpha 2-binding sites in the medulla oblongata. In tree shrews, alpha 2-adrenoceptors can be autoradiographically quantified in regions which are not labeled in the rat, although former data predicted the existence of such receptors, e.g., in the area of the adrenaline cell group C1.

Animals↗
PubMed1990

The initiation of translation in E. coli: apparent base pairing between the 16srRNA and downstream sequences of the mRNA.

Bacteriophage T7's gene 0.3, coding for an antirestriction protein, possesses one of the strongest translation initiation regions (TIR) in E. coli. It was isolated on DNA fragments of differing length and cloned upstream of the mouse dihydrofolate reductase gene in an expression vector to control the translation of this gene's sequence. The TIR's efficiency was highly dependent on nucleotides +15 to +26 downstream of the gene's AUG. This sequence is complementary to nucleotides 1471-1482 of the 16srRNA. Similar sequences complementary to this rRNA region are present in other efficient TIRs of the E. coli genome and those of its bacteriophages. There seems to be a correlation between this sequence homology and the efficiency of the initiation signals. We propose that this region specifies a stimulatory interaction between the mRNA and 16srRNA besides the Shine-Dalgarno interaction during the translation initiation step.

Bacterial Proteins↗
PubMed1990

Occupational performance of a paced secondary task under conditions of sensory deprivation. I. Heart rate changes in train drivers as a result of monotony.

Twelve experienced train drivers were asked to operate the train function safety circuit (SIFA)--a paced secondary motor task which is expected to guarantee the driver's fitness for service on engines of the German Federal Railway--under laboratory conditions of extreme monotony. In spite of massive decreases in vigilance as shown by theta-activity in the EEG, all subjects were able to operate the device without major errors. A prerequisite for adequate performance is an EEG-defined arousal reaction, which is synchronized with the SIFA cycles. For 7 subjects the time-related cross-correlation coefficients between SIFA operation, alpha-activity (indicative of alertness), theta-activity (indicative of reduced vigilance), and heart rate were calculated. The central-nervous arousal found in the EEG corresponded to distinct increases in heart rate. The moments of SIFA operation after phases of light sleep correlated significantly and positively with increases in heart rate. These increases constituted a physiological overcompensation as far as the physical readiness was concerned which reached its peak after the task had been performed satisfactorily, constituting additional and superfluous occupational stress. The results of this study indicate clearly that monotony stress is not a result of occupational monotony itself, but the result of the physiological effort which is required in order to regain a level of alterness which allows adequate performance under monotonous conditions.

Electroencephalography↗
PubMed1990

Occupational performance of a paced secondary task under conditions of sensory deprivation. II. The influence of professional training.

Twelve truck drivers operated the train function safety circuit (SIFA), a paced secondary task used as a job monitor on German railways engines, under laboratory conditions of extreme monotony, in a comparison with 12 train drivers who were well acquainted with SIFA. Alertness was determined by means of EEG evaluations. Heart rate was monitored as the parameter for physical load, and the precoded SIFA tasks as the active response parameter. In spite of significantly more frequent and more distinct decreases in alterness, the SIFA-trained subjects (TS) performed better. Nine out of 12 TS reached the stage of light sleep at least once during the experimental run, as compared to 4 out of 12 untrained subjects (US). Nevertheless, the ratio acoustic warnings/occurrence of light sleep was significantly lower in TS (P less than 0.01), and there were three operational errors (equivalent to emergency braking) in US. Whereas US received fewer acoustic warnings in the stages of slightly reduced altertness, this trend was reversed as soon as low frequency theta-activity appeared in the EEG. A time-related calculation of the cross-correlation coefficients between SIFA operation, alpha-activity, theta-EEG-activity, and heart rate showed that timing of SIFA operation interrupting phases of light sleep correlated significantly and positively with increases in heart rate. The results suggest that a mechanism of rhythmic central nervous arousal interrupting phases of decreased alertness/drowsiness can be learned, whereas the physiological consequences of the effort to prevent the deterioration of performance under conditions of monotony are not reduced by professional adaptation.

Automobile Driving↗
PubMed1990

Psychosocial stress affects pineal function in the tree shrew (Tupaia belangeri).

Using a recently developed commercially available radioimmunoassay the concentration of the principal melatonin metabolite 6-sulfatoxymelatonin (aMT6s) in the morning urine of male tree shrews was determined. Chronic social confrontation elicited a drastic increase of aMT6s excretion in subordinate tree shrews, whereas there was a tendency to reduced excretion of the melatonin metabolite in dominant animals. These results substantiate the function of the pineal gland in transforming stimuli from the social environment to endocrine information and, therefore, are indicative for the relevant role the gland may play in the physiological reactions to chronic psychosocial stress.

Agonistic Behavior↗