Recently, the human orthologue to the cell cycle checkpoint genes rad17 (Schizosaccharomyces pombe) and RAD24 (Saccharomyces cerevisiae), called HRAD17, has been isolated and localized to chromosome 4. Independently, we have isolated the HRAD17 transcript and mapped it to chromosome 5q13 between the CCNB1 and BTF2p44cen genes. Furthermore, we have identified the complete exon-intron structure of HRAD17. The gene is organized into 14 exons, the translation initiation site lies within exon 2, and the stop codon within exon 14. Two further HRAD17 pseudogenes, HRAD17P1 and HRAD17P2, were identified on chromosomes 7p21 and 13q14.3, respectively, encompassing exons 3-14 and bearing 84% and 93% homology, respectively. Additionally, we have isolated the coding region of the mouse orthologue, Mrad17, and mapped it on chromosome 13 between Ccnb1 and Btf2p44, the same two genes between which it maps in human. The predicted Mrad17 polypeptide encompasses 687 amino acids and shows 89% similarity to HRAD17. Both genes are most highly expressed in testis compared to all other tissues, as shown by Northern blot hybridization. Histological studies, based on in situ hybridization with radioactively labeled antisense HRAD17 riboprobes, showed a strong expression within the germinal epithelium of the seminiferous tubuli in normal testis whereas in testicular tumors (seminomas) only weak, diffuse signals were seen. In light of the known function of the yeast orthologue at meiotic and mitotic checkpoints, as well as the strong expression in testis and weak expression in seminomas, we suggest a putative involvement of HRAD 17 in testicular tumorigenesis.

Animals↗

PubMedDate not supplied

Solvent optimization for the direct extraction of opiates from hair samples.

The hair samples of six opiate addicts, who died after heroin overdose, were investigated by gas chromatography-mass spectrometry after extraction with 10 solvents differing in polarity and hydrophilicity in an ultrasonic bath. Morphine, 6-monoacetylmorphine (MAM), and codeine were detected in all cases, and heroin was detected in four cases. With toluene, which is hydrophobic, almost no extraction occurred, and with the nonprotic solvents dioxane, acetonitrile, acetone, and dimethyl sulfoxide, only a relatively low extraction rate was found. The yield increased in the series of alcohols from n-propanol to isopropanol to ethanol to methanol. Water proved to have almost the same extraction capability as methanol. Using equal conditions, the extraction rate of the opiates decreased in the following order: heroin > MAM > morphine approximately codeine. Addition of 1% acetic acid or 1% triethylamine to methanol led to a decrease in the heroin yield and an increase in the morphine yield. The results are discussed in terms of the binding between the hair matrix and the drug, the penetration of the solvent into the hair, and the solubility of the drug in the solvent.

Adult↗

PubMedDate not supplied

[Breath condensate--a medium obtained by a noninvasive method for the detection of inflammation mediators of the lung].

Collection of exhaled condensate (freezing of expired air under conditions of spontaneous breathing) is a non-invasive method permitting the collection of material originating from the lung and the lower respiratory tract so that it can be used for diagnostic examinations. In order to be able to evaluate the diagnostic evidence of exhaled condensate samples in cases of respiratory disease of the calf, leukotriene B4 (LTB4) in bovine exhaled condensate was determined. The influence of the breathing pattern and body temperature on the quantity of condensate to be collected was tested in a total of 49 exhaled condensate samples. It became obvious that the exhaled condensate quantity obtained per time unit is dependent on the ventilation volume per time unit (minute volume). In exhaled condensate samples from 35 clinically healthy calves, LTB4 concentrations of less than 250 pg/mL exhaled condensate were detected. A total of 14 exhaled condensate samples from 7 calves was analyzed before and after experimental respiratory infection with Pasteurella multocida D. In parallel to the analysis of LTB4 in exhaled condensate, the lung function of the calves was examined by means of impulse oscilloresistometry. The increase of LTB4 in the exhaled condensate post infection correlated significantly (p < or = 0.05) with decreases of respiratory reactance. The determination of LTB4 concentrations in exhaled condensate seems to be suitable, in principle, for the detection of inflammations in the respiratory system of the calf. Further studies are needed for the evaluation of the diagnostic validity of the method.

Animals↗

PubMedDate not supplied

Comparing opinions about mental illness for hospital attendants and practical nursing students.

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Attitude of Health Personnel↗