Rapid detection of human cytomegalovirus DNA in peripheral blood leukocytes of viremic transplant recipients by the polymerase chain reaction.

Peripheral blood leukocyte samples (n = 458) of 24 bone marrow transplant and 52 kidney transplant patients were examined weekly for the presence of human cytomegalovirus (HCMV) using an improved culture technique (DEAFF; detection of early antigen fluorescent foci). In total 5 (21%) bone marrow transplant and 11 (21%) kidney transplant patients developed a viremia. Patients' samples were investigated for the presence of HCMV DNA using an in vitro DNA amplification technique, the polymerase chain reaction (PCR). From the statistically evaluable viremic patients (n = 13), 110 blood samples were analyzed. In 5 of these patients, the DEAFF and PCR led to identical results. In 8 patients however the PCR was more sensitive, i.e. HCMV DNA was detected for a longer period of time. Applying statistical analysis using the McNemar test, this result was significant (P less than 0.05). The PCR applied on leukocyte samples did not detect HCMV DNA in viruric patients without viremia. Moreover, the current PCR never led to positive results with peripheral blood leukocyte samples of healthy seropositive or seronegative controls. Since the PCR can be performed in 6 hr, this technique will contribute to rapid detection of HCMV DNA in peripheral blood leukocytes and therefore to optimal clinical management of HCMV-infected transplant recipients.

Expression of c-sis and platelet-derived growth factor in in vitro-transformed glioma cells from rat brain tissue transplacentally treated with ethylnitrosourea.

Long-term culturing of brain cells from neonatal BD-IX rats after transplacental treatment with N-ethyl-N-nitrosourea (ENU) results in malignantly transformed cells after a lag period of about 250 days. During culturing, the brain cells undergo a sequence of morphological changes. We examined oncogene expression in cultured cells from ENU-treated animals and found that transformed glioma cells differ from premalignant glial cells by containing high levels of c-sis transcripts. We also report that the transformed cells synthesize functional platelet-derived growth factor. Because glial cells have receptors for platelet-derived growth factor, we propose that an autocrine mechanism plays an important role in ENU-induced brain tumorigenesis.

An antiserum against a glycoprotein functional in flagellar adhesion between Chlamydomonas eugametos gametes.

Chlamydomonas eugametos gametes agglutinate sexually by their flagellar surfaces. The agglutination factor on mating type minus (mt-) gametes is thought to be a glycoprotein named PAS-1.2. To test this idea, an antiserum was raised against purified PAS-1.2, which reacted with isolated PAS-1.2 (immunoprecipitation tests) and blocked the ability of isolated PAS-1.2 to induce sexual twitching in mt+ gametes. When tested with living cells, the antiserum specifically agglutinated mt- gametes and induced a reaction resembling twitching. Mt+ flagella were shown to bind the antiserum (indirect immunofluorescence) but much less than mt- gametes. Mt- gametes pretreated with Fab fragments of the antiserum were unable to reproduce sexually, while treated mt+ gametes were unaffected. This effect presumably results from the ability of the serum to block mt- sexual agglutination, for mt- isoagglutinin was completely inactivated by the serum, while mt+ isoagglutinin was unaffected. It is therefore argued that PAS-1.2 is the in vivo mt- agglutination factor. However it is shown that the antiserum was able to react in vitro not only with PAS-1.2 but with several other proteins in both mt- and mt+ flagella.