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Biomedical subjects

J Kopp

Publications and source records attributed to J Kopp.

102 records · Page 6Linked to original sources

A simple method for cloning and replica plating of mammalian cells using multicellular spheroids.

V 79/4 Chinese hamster cells or HeLa cells grow in Eagle's MEM supplemented with 25 microgram/ml dextran sulphate to form clonal multicellular spheroids. These cell clones, consisting of 5-10(2) cells, are easy to separate, to transfer from one culture vessel into another and grow as normal monolayer colonies on Dederon cloth circles after subculture in Eagle's MEM without dextran sulphate. A simple replica technique is described by which 500 clones can be transfered onto at least 3 replica cloth circles, 10 cm in diameter, with a replica plating efficiency of approximately 100%.

Animals↗

Measurements of radiation exposure in civil aircraft.

The radiation exposures on 12 flights of German airlines were measured with an active dosemeter based on two silicon semiconductors. The dependence on the date, altitude and route of the flights was studied. Measured dose rates and preliminary dose equivalent rates of the individual flights are given and compared with model calculations.

Aerospace Medicine↗

Liver fibrogenesis due to cholestasis is associated with increased Smad7 expression and Smad3 signaling.

BACKGROUND/AIMS: Profibrogenic TGF-beta signaling in hepatic stellate cells is modulated during transdifferentiation. Strategies to abrogate TGF-beta effects provide promising antifibrotic results, however, in vivo data regarding Smad activation during fibrogenesis are scarce. METHODS: Here, liver fibrosis was assessed subsequent to bile duct ligation by determining liver enzymes in serum and collagen deposition in liver tissue. Activated hepatic stellate cells were identified by immunohistochemistry and immunoblots for alpha smooth muscle actin. Cellular localization of Smad3 and Smad7 proteins was demonstrated by immunohistochemistry. RTPCR for Smad4 and Smad7 was conducted with total RNA and Northern blot analysis for Smad7 with mRNA. Whole liver lysates were prepared to detect Smad2/3/4 and phospho- Smad2/3 by Western blotting. RESULTS: Cholestasis induces TGF-beta signaling via Smad3 in vivo, whereas Smad2 phosphorylation was only marginally increased. Smad4 expression levels were unchanged. Smad7 expression was continuously increasing with duration of cholestasis. Hepatocytes of fibrotic lesions exhibited nuclear staining Smad3. In contrast to this, Smad7 expression was localized to activated hepatic stellate cells. CONCLUSIONS: Hepatocytes of damaged liver tissue display increased TGF-beta signaling via Smad3. Further, negative feedback regulation of TGF-beta signaling by increased Smad7 expression in activated hepatic stellate cells occurs, however does not interfere with fibrogenesis.

Animals↗

Single-cell suspensions of cultured human keratinocytes in fibrin-glue reconstitute the epidermis.

To overcome common disadvantages of standard cultured epidermal sheet grafts (CEG) we have developed a new technique of transplanting cultured human keratinocytes suspended as single cells in a fibrin-glue matrix (Keratinocyte-fibrin-glue suspension-KFGS). In an athymic mouse model with reproducible standardized full thickness wounds this new technique was compared directly to CEG. Reepithelialization was similar in both groups, but reconstitution of the dermo-epidermal junction zone, as shown by electron microscopy and immunohistochemistry was significantly enhanced by the fibrin-glue suspension technique. The new KFGS technique is earlier available than sheet grafts, is able to transfer actively proliferative single keratinocytes, and simplifies the application.

Animals↗

Increase in Mycobacterium avium complex isolations among patients admitted to a general hospital.

In early 1979, an official of an Illinois hospital reported an increase in the number of patients from whom Mycobacterium avium complex recently had been recovered. Over the preceding 3 years specimens from a total of 51 patients were culture positive for M. avium complex: 7 in 1976, 8 in 1977, and 36 in 1978. Nine of 10 serotyped isolated were serotype 8. The increase was not attributable to an increase in the number of mycobacterial cultures performed. No other area hospitals had similar increases in rates of recovery of M. avium complex. Patients with M. avium complex were significantly more likely than patients with other mycobacteria to have been residents of the city where the hospital is located. The distribution of abnormalities in patients' chest films differed significantly between patients with M. avium complex in 1978 and patients with M. avium complex in 1976-77; in 1978, patients although equally likely to have infiltrates, nodules, or cavities, were more likely to have no abnormalities or abnormalities consistent with chronic obstructive pulmonary disease, and less likely to have other abnormalities. The data suggest that the increased rate of recovery of M. avium complex from patients could not be attributed to ascertainment bias or laboratory variation but may be due to an increase in the incidence of disease or colonization among persons living in the community where the hospital is located.

Adolescent↗

Paracrine stimulation of keratinocytes in vitro and continuous delivery of epidermal growth factor to wounds in vivo by genetically modified fibroblasts transfected with a novel chimeric construct.

BACKGROUND: Growth factors play an important role in tissue repair. While the effectiveness of growth factor therapy in animal wound healing models and limited human clinical trials has been demonstrated, the ideal method for their administration to the wound remains unclear. Experimental data suggest that the continuous presence in the early stages of wound repair is beneficial. MATERIALS AND METHODS: We have constructed a novel chimeric expression plasmid in which the biologically active portion of the human epidermal growth factor (EGF) gene is fused in-frame to the human granulocyte colony-stimulating factor signal sequence. RESULTS: Clonally selected human fibroblasts transfected with this construct secrete biologically active EGF. After the transplantation of irradiated gene-transfected fibroblasts suspended in fibrin glue to murine full-thickness wounds, EGF can be demonstrated for at least seven days in the wounds, slowly decreasing from initially 470 pg/ml to 140 pg/ml on day 7. No EGF was found in the wound at day 14. CONCLUSIONS: A single application of irradiated EGF genetransfected fibroblasts to wounds can thus continuously deliver the transgene in vivo and could be used to administer drugs to the wound bed during the crucial first seven days of wound-healing.

Amino Acid Sequence↗