[Vernix aspiration in newborn].
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In order to analyse the evolution of the sensitivity to antibiotics of same strains with nosocomial potential such as Klebsiella isolated in a hospital, we took into study a number of 976 samples, collected in a new-born care department of the "Dr. D. Popescu" hospital Timişoara. The study took place between November-December 2002. The collected samples were pharyngeal swabs, nasal swabs, gastric aspirates, ocular and umbilical secretions, vernix, urines, faeces and blood. From all these samples 803 strains with nosocomial potential were isolated, 84 strains being Klebsiella spp. Most of the isolated strains presented multiple phenotypes of resistance to antibiotics.
OBJECTIVE: To explore the preliminary methods of in vitro isolation, culture and identification of sebocytes and eccrine sweat gland cells from human fetal skin. METHODS: Human fetal skin was digested with dispase or type II collagenase, and then by micro - sieving to isolate human sebaceous gland and eccrine sweat gland cells. DMEM/F12 (1: 1) was used as the basic culture medium, supplemented with fetal bovine serum, recombinant human epidermal growth factor, L-glutamine, Hydrocortisone, choleratoxin, penicillin and streptomycin as the medium for sebocytes, or fetal bovine serum, recombinant human epidermal growth factor, triiodothyronine, hydrocortisone, insulin, transferrin, sodium selenite to the medium for eccrine sweat gland duct cells. Primary cultures and subcultures were incubated at 37 degrees C in humidified atmosphere of 5% CO2/95% oxygen. Cell morphology was observed by inverted phase contrast microscopy, and the cultured cells were identified with cell clone efficiency determination. The cultured sebocytes were identified with oil red staining and CK4.62, Epithelia Membrane Antigen (EMA) immunohistochemistry staining. The cultured eccrine sweat gland duct cells were identified with CK7, CK19 immunohistochemistry staining. RESULTS: The isolated sebocytes and eccrine sweat gland cells from human fetal skin could grow by adhering to the wall and proliferate in vitro. The cell clone efficiency of human fetal sebocytes was 2.7%, which was obviously lower than that of human fetal keratinocytes (8.0%, P < 0.01). There was no obvious difference in the cell clone efficiency between human fetal eccrine sweat gland cells (7.3%) and human fetal keratinocytes (7.7%, P > 0.05) . The results of oil red staining indicated that a small quantity of lipid droplets in sebocytes, and immunohistochemistry staining of CK4.62, EMA were positive in subculture sebocytes. The immunohistochemistry staining of CK7, CK19 was positive in subculture eccrine sweat gland duct cells. CONCLUSION: In vitro cultured human fetal sebocytes and eccrine sweat gland duct cells displayed the markers and biological characteristics of epithelial lineage, but human fetal sebocytes proliferated more
The relationship between ultrasonographic detection of fetal vernix and visual assessment of amniotic fluid (AF) and fetal pulmonary maturity evaluated by the "shake test" was studied in 73 high-risk patients undergoing amniocentesis for obstetrical indications. The AF was classified as type I, type II, and type III, depending on the subjective amount of vernix detected. Ultrasound predicted correctly 7 out of 11 cases of type I AF (64%), 14 out of 22 cases of type II AF (64%), and 38 out of 40 cases of type III AF (95%). When ultrasonography detected type I AF the "shake test" was negative in all cases, whereas if type II or type III were found, the "shake test" was positive in 40% and 86%, respectively. Our results suggest that sonographic detection of type I AF is associated with immaturity, thus amniocentesis should be avoided in such cases. Conversely, if a type III AF is detected a mature fetus is found in almost 90% of the cases. The observation of AF detecting fetal vernix during third trimester sonography is a simple manoeuver that can help to avoid the use of amniocentesis, an invasive procedure associated with some serious fetal and maternal complications.
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