Biomedical subjects

Monika Egerbacher

Publications and source records attributed to Monika Egerbacher.

8 recordsLinked to original sources

PubMed2005

HIV-1 Rev can specifically interact with MMTV RNA and upregulate gene expression.

We present evidence that the HIV-1 Rev protein can heterologously regulate expression of the simple beta retrovirus mouse mammary tumour virus (MMTV). Up to 10-fold upregulation was seen in a functional assay system when specific MMTV sequences were substituted for the HIV-1 Rev responsive element (RRE). RNA gel shift analysis showed that purified recombinant Rev could specifically bind to MMTV unique region 3 prime (U3) RNA and that these sequences could compete for wild-type Rev-RRE binding approximately 20-fold more efficiently than a non-specific competitor RNA. Using a combination of in silico and deletion mutation analyses, it was not possible to define any single specific secondary structure responsive to Rev, suggesting that a structure or combination of structures that only form in the context of the complete U3 transcript is/are required to interact with Rev. Taken together, these results suggest that HIV-1 Rev can directly bind to MMTV RNA as well as mediate upregulation of MMTV gene expression.

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PubMed2005

Development of a transgenic mouse model susceptible to human coronavirus 229E.

Human coronavirus (HCoV) 229E is a group 1 coronavirus and is specific to humans. So far, no animal model is available to study the pathogenesis of infection by HCoV-229E. We show here that the expression of aminopeptidase N (APN, also termed CD13), the receptor for HCoV-229E, is required but not sufficient to confer susceptibility in vivo. HCoV-229E infection was facilitated by crossing APN transgenic mice into signal transducers and activators of transcription (Stat) 1 null mice and by adaptation of HCoV-229E to grow in primary APN transgenic, Stat1 null fibroblasts. Double transgenic mice allow the study of human coronavirus group 1 infections in an animal model, in particular, viral tropism, replication, recombination, and spread in an immunocompromised situation. Furthermore, these mice provide an important tool for the evaluation of biosafety and efficacy of coronavirus-based vectors.

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PubMed2005

Prevalence and radiologic and histologic appearance of vesicourachal diverticula in dogs without clinical signs of urinary tract disease.

OBJECTIVE: To determine prevalence and radiologic and histologic appearance of vesicourachal diverticula in dogs without clinical signs of urinary tract disease. DESIGN: Original study. ANIMALS: 50 dogs between 4 months and 17 years old representing 22 breeds that had been euthanatized for unrelated reasons; none of the dogs had a history or clinical signs of urinary tract disease. PROCEDURE: Retrograde positive-contrast radiography was performed, and radiographs were examined for macroscopic diverticula. Necropsy specimens from the urinary bladder vertex were examined by means of light microscopy for diverticula and signs of inflammation. RESULTS: 17 of the 50 (34%) dogs had vesicourachal diverticula, and 1 additional dog had a urachal cyst. Fifteen of the 17 diverticula were macroscopic; surface area of the diverticulum could be measured radiographically in 13 of these dogs and ranged from 1 to 90 mm2. The remaining 2 diverticula were microscopic. Sixteen diverticula were intramural and 1 was extramural. Light microscopic signs of bladder wall inflammation could be detected in 5 dogs, 4 of which had macroscopic diverticula. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a high percentage of dogs without clinical signs of urinary tract disease may have vesicourachal [corrected] diverticula. Further studies are needed to determine the clinical relevance of vesicourethral diverticula in dogs.

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PubMed2005

Skin plugs in phlebotomy puncture for blood donation.

Contamination at the site of the donor's skin may occur despite proper disinfection, because pathogens in deeper regions (such as pores) may not be eliminated by skin disinfection. It is suspected that the cannula detaches fragments of tissue when it penetrates the skin; the tissue fragments may reach blood products and release pathogens there. In the present study we punctured piglet skin with cannulas commonly used for blood donation and performed histological as well as cytological investigations of the lavage fluid in the cannula to identify superficial skin cells and skin plugs. Histological specimens of the pierced skin showed frayed puncture sites with loosely attached tissue fragments. In the lavage fluid of the cannula, a collection of epidermal cells was found in one of 150 punctures. Our results confirm that the phlebotomy cannula may cause superficial tissue fragments to be punched out of the donor's skin during blood donation. This fact should be taken into account when devising methods to reduce bacterial contamination in blood products.

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PubMed2005

A 470 bp WAP-promoter fragment confers lactation independent, progesterone regulated mammary-specific gene expression in transgenic mice.

The ability of a 470 bp sub-fragment of the murine whey acidic protein (WAP) promoter in the context of a retroviral expression plasmid to direct gene expression to mammary epithelial cells was analysed in a number of independent transgenic mouse lines. In contrast to previous findings with the genuine 2.5 kb promoter fragment, our studies revealed a highly mammary gland-specific expression detectable only in non-lactating animals. This suggested a mainly progesterone-regulated activity of the short fragment. Therefore, transgene expression was examined in the progesterone-determined estrous cycle and during pregnancy. In accordance with in vitro data from stably transfected cell lines, in both situations expression was upregulated at stages associated with high progesterone levels. Taken together these data provide deeper insight into WAP-promoter regulation and stress the usefulness of the shortened fragment for a lactation independent mammary-targeted expression.

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PubMed2004

Decrease in laminin content and protein excretion rate after five sixths nephrectomy and low-dose irradiation in the rat.

The effect of low-dose irradiation on laminin distribution and urine protein excretion in the remnant rat kidney has been studied. The rat remnant kidney formed after 5/6 nephrectomy is an experimental model of chronic renal failure. In the remnant kidney, focal segmental glomerulosclerosis is developed characterized by focal or segmental sclerosis in glomeruli, alterations in the tubules and thickening of the glomerular basement membrane. Low dose irradiation has been presumed to suppress sclerotic processes. In this study 24 male Wistar rats were subdivided into the nephrectomized group, nephrectomized and irradiated groups (1 or 3 Grey), and healthy control group. Animals were sacrificed at 2, 4 and 8 weeks after beginning the experiment. Laminin immunohistochemical staining was found along the tubular and glomerular basement membranes in all experimental groups, but with varying intensity. Laminin content in the basement membranes was decreased in early stages (week 2), especially after irradiation followed by increase during the later stages with relatively high levels at the end of the experiment (week 8). Irradiation at a dose of 3 Grey decreased protein excretion compared to the nephrectomized rats at all stages, while 1 Grey dose was ineffective. Based on decreased proteinuria we conclude that moderate low-dose irradiation has beneficial effects on the rat remnant kidney and that laminin in basement membranes is probably not the most crucial component in regulating membrane permeability.

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PubMed2002

Primary cell culture and morphological characterization of canine dermal papilla cells and dermal fibroblasts.

Skin biopsies were taken from female dogs, the primary hair follicles isolated and the dermal papilla dissected. After incubation in supplemented Amniomax complete C100 medium in 24-well culture plates, the dermal papilla cells (DPC) grew to confluence within 3 weeks. Thereafter, they were subcultivated every 7 days. Dermal fibroblast (DFB) cultures were established by explant culture of interfollicular dermis in serum-free medium, where they reached confluence in 10 days. They were subcultivated every 5 days. For immunohistochemistry, cells were grown on cover slips for 24 h, fixed and stained with antibodies against collagen IV and laminin. DPC showed an aggregative growth pattern and formation of pseudopapillae. Intensive staining for collagen IV and laminin could be observed until the sixth passage. DFB grew as branching, parallel lines and showed only weak staining for collagen IV and laminin.

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PubMed2002

Immunohistochemical localization of androgen and oestrogen receptors in canine hair follicles.

Skin biopsies from seven different body sites were obtained from 21 dogs of different breeds (short, normal, long hair), which were presented for euthanasia. Commercially available polyclonal antibodies were used for the immunohistochemical detection of androgen and oestrogen receptors. Both receptors showed a similar distribution in canine skin, with specific intranuclear staining. In the epidermis, the percentage of androgen receptor (AR)-positive cells, but not that of oestrogen receptor (ER)-positive cells, was significantly higher in samples from the thorax and the flank. In the dermal papilla, the percentage of ER-positive (but not AR-positive) cells was significantly lower in biopsies from the flank. No significant difference was found for both receptors between the locations in the outer root sheath, among the three different hair types, between sex and between intact and castrated dogs.

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