Virus isolation from and identification of HTLV-III/LAV-producing cells in brain tissue from a patient with AIDS.

Primary cultures from a brain biopsy specimen of a human T-cell lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV) seropositive patient with progressive dementia contained small numbers of monocytoid cells and showed reverse transcriptase activity that persisted for as long as 100 days. Electron microscopy of these cells revealed the presence of HTLV-III/LAV virions. Subcultured cells removed from primary cultures by trypsinization were nonspecific esterase negative and did not express virus or show evidence of HTLV-III/LAV proviral sequences, while those remaining in the original flasks were nonspecific esterase positive and continued to produce virus. Virus from primary cultures was transmitted to peripheral blood-derived monocyte-macrophages and T cells. Virus production in T-cell cultures was transient while the monocyte-macrophages, like the primary cultures, produced virus for at least 120 days. Infection of several brain-derived cells with this and another HTLV-III/LAV isolate failed to demonstrate virus replication. These results indicate that the HTLV-III/LAV-infected cells recovered from the brain of this patient are cells of the mononuclear phagocyte series.

The role of mononuclear phagocytes in HTLV-III/LAV infection.

Cells with properties characteristic of mononuclear phagocytes were evaluated for infectivity with five different isolates of the AIDS virus, HTLV-III/LAV. Mononuclear phagocytes cultured from brain and lung tissues of AIDS patients harbored the virus. In vitro-infected macrophages from the peripheral blood, bone marrow, or cord blood of healthy donors produced large quantities of virus. Virus production persisted for at least 40 days and was not dependent on host cell proliferation. Giant multinucleated cells were frequently observed in the macrophage cultures and numerous virus particles, often located within vacuole-like structures, were present in infected cells. The different virus isolates were compared for their ability to infect macrophages and T cells. Isolates from lung- and brain-derived macrophages had a significantly higher ability to infect macrophages than T cells. In contrast, the prototype HTLV-III beta showed a 10,000-fold lower ability to infect macrophages than T cells and virus production was one-tenth that in macrophage cultures infected with other isolates, indicating that a particular variant of HTLV-III/LAV may have a preferential tropism for macrophages or T cells. These results suggest that mononuclear phagocytes may serve as primary targets for infection and agents for virus dissemination and that these virus-infected cells may play a role in the pathogenesis of the disease.

Effects of in vitro infection of mouse glial and neuroblastoma cells with the scrapie agent.

Mouse glial and neuroblastoma cells were infected with the mouse adapted strains C506 and 139A of scrapie agent. Lysates of the in vitro infected cells (from the 3rd to the 16th passage) intracerebrally inoculated into CD-1 mice, caused the development of a neurological disease, with characteristic signs of scrapie. Morphological changes in scrapie-infected neural cells were observed after about fifteen in vitro passages. In liquid medium, the cloning efficiency of these cells increased. They acquired the capacity to form large tridimensional colonies in agar. Heating the infectious brain extracts at 60 and 80 degrees C for 30 minutes did not inhibit these changes thus showing the involvement of the thermoresistant scrapie agent. Supernatants of scrapie-infected glial cells promoted colony formation in liquid medium with different types of normal cells. Analysis of supernatants of scrapie-infected mouse neuroblastoma cells showed a profound modification of neurotransmitter metabolism.

In vitro propagation of the scrapie agent. I. Transformation of mouse glia and neuroblastoma cells after infection with the mouse-adapted scrapie strain c-506.

Seven cell lines including glia cells from mouse brains and mouse neuroblastoma cells were infected with the mouse-adapted scrapie strain c-506. During the early in vitro passages, a stimulation of growth was already observed but cellular morphology and differentiation did not alter. Later on, after 12-16 passages, six of the seven infected lines displayed cell proliferation and morphological alterations, suggesting an in vitro morphological transformation. At this stage, differentiation was no longer observed in the scrapie-infected neuroblastoma cells and all the scrapie-infected cells formed two to four times more colonies in liquid medium than the controls, and developed large tridimensional colonies in agar. The part played by the scrapie agent in these changes is discussed.

[Lymphography as an investigation method for bladder cancer in adults]. / Intérêt de la lymphographie dans le bilan pré-thérapeutique des cancers vésicaux de l'adulte.

The use of lymphography as an investigational method in bladder cancer is a controversial subject. Results of a personal series of 90 cases, including 50 that were histologically confirmed, are compared with those reported in the published literature. Results were fairly different from those previously described, as though there was a lower sensitivity (43 as against 85 p. cent in the literature), specificity was much higher (100 as against 94 p. cent) as well as positive predictive value (100 as against 69 p. cent). This improvement was mainly due to the use of Piver and Wallace's assessment criteria, and results should be improved still further in the future by more frequent use of needle puncture cytology. Even with the increasing employ of the scanner, lymphography still remains a precise method for investigating bladder cancer.

[In vitro modification of the morphology and the growth of cells infected with scrapie (author's transl)]. / Modifications in vitro de la morphologie et de la croissance de cellules infectées par l'agent scrapie.

Seven cell lines originated either in brains or in neuroblastomas of Mice, were infected with Scrapie. After 12 to 16 in vitro passages, 6 lines out of 7 showed changes of their morphology, and of their growth, resembling those occurring in the course of a malignant transformation. The Scrapie infected cells acquired the capacity to form 2 to 4 times more colonies in liquid medium than the controls, and to develop large tridimensional colonies in semisolid medium. The role of Scrapie in these changes is discussed.

[Trials of in vitro propagation of the scrapie agent in mouse nerve cells]. / Essais de propagation in vitro de l'agent scrapie dans des cellules nerveuses de souris.

We attempted to propagate the Scrapie agent in vitro in glia and neuroblastoma cells of Mice. Four out of seven assays of infection were positive, i.e. after several passages in vitro yielding at most a 10(9) fold final dilution of the original material, the extracts of each of the four cultures, when injected intracerebrally into CD1 Mice, produced a deadly disease displaying the clinical and pathological signs characteristic of Scrapie.

Metabolism of benzo[a]pyrene by brain microsomes of fetal and adult rats and mice. Induction by 5,6 benzoflavone, comparison with liver and lung microsomal activities.

Using brain, lung and liver microsomes as the enzyme source in in vitro assays, benzo[a]pyrene (B[a]P) metabolism was studied in fetuses and dams of mice (C57B1/6) and rats (WAG). Separation and quantitation of B[a]P metabolites were performed by h.p.l.c. Microsomal preparations were tested for cytochrome P-450 dependent O-dealkylation of 7-ethoxycoumarin and epoxide hydrolase activities. Another parameter measured included the conjugation of 1-chloro-2,4 dinitrobenzene to glutathione by cytosolic glutathione-S-transferase activity. The induction of B[a]P metabolism was studied after treatment of animals with 5,6-benzoflavone (BF). Mixed function oxygenase, epoxide hydrolase and glutathione-S-transferase activities were transplacentally inducible after dams were treated with BF. Metabolic activation of B[a]P by fetal brain microsomes was lower in both species than that by fetal lung and liver microsomes, but it was higher in fetuses than in adults. All metabolites of B[a]P increased after BF treatment; the production of 7,8-dihydro-7,8-dihydroxybenzo[a]pyrene (7,8-dihydrodiol B[a]P) was higher in brain microsomes from BF-treated rats than that in mice. In stimulated rats, the formation of 7,8-dihydrodiol B[a]P by fetal brain microsomes were higher than that by fetal lung microsomes, whereas in mice, the opposite was observed. These data suggest that initiation could occur in utero, and partially explain the species-specific differences in susceptibility to transplacental tumorigenesis by polycyclic aromatic hydrocarbons by differences in biotransformation in the target organ.

Lymphography in childhood rhabdomyosarcomas.

Forty-one children with subdiaphragmatic rhabdomyosarcoma underwent bipedal lymphography. Twenty-two (53.5%) of the lymphograms were interpreted as being positive. In our series, the lower limbs were the most common primary site of involvement, were more frequently involved by the alveolar histologic subtype which carries a poor prognosis, and were associated with a higher incidence of lymph node metastases. Positive lymphographic findings in this group of children were similar to those seen in both adults and children with other solid tumors, i.e., the presence of discrete lymph node filling defects. However, in 3 cases, abnormalities more characteristic of lymphoma were identified. Evaluation of lymph node metastases as demonstrated by lymphography has prognostic significance.

Normal and benzo(a)pyrene-transformed fetal mouse brain cell. I. Tumorigenicity and immunochemical detection of glial fibrillary acidic protein.

Primary cultures of whole brain and cortex cells origination from 14-day-old A/Jax or C3H mouse fetuses were treated with benzo(a)pyrene (B(a)P) for 24 h. After 7 to 8 passages a malignant transformation was observed in the chemically treated whole brain and cortex cultures. Control cultures of cortex remained non-transplantable during the whole experiment (up to 14 passages) whereas in the control cultures originating from whole brain a spontaneous transformation appeared after 11 passages. With horseradish peroxidase-labelled antibody, the specific glial fibrillary acidic protein (GFAP) was detected in both control and transformed total brain and cortex cultures, and in the tumors initiated by the in vitro transformed cells. This finding shows that glialike cells persisted after a long in vitro maintanance and transformation.

Normal and benzo(a)pyrene-transformed fetal mouse brain cells. II. Ultrastructural study.

An ultrastructural study was performed on normal and Benzo(a)-pyrene(B(a)P)-transformed fetal mouse brain cells. Early subcultures of a strain initiated from whole brain presented three cell types in vitro: astroglial, poorly differentiated glial, and spongioblastic types. After B(a)P-treatment, there was an exclusive transformation and the growth of neuroglia sometimes without gliofibrillary maturation, but with the presence of glial fibrillary acidic protein (GFAP) in the cytoplasm. Early subcultures of another strain initiated from cortex only presented poorly differentiated neuroglial cells. After transformation, cell maturation as evidenced by gliofibrillogenesis and GFAP production by these cells was observed. In both cases, the potentiality of glial differentiation after in vitro malignant transformation by a chemical carcinogen seemed preserved.

[In vitro malignant transformation of hamster lung cells by 2-chlorobutadiene]. / Transformation in vitro de cellules de poumons de jeunes hamsters par le chloro-2 butadiène (CB).

Normal Hamster lung cells from an established line were treated with 1-500 microgram . ml-1 2-chlorobutadiene. Those treated with 1 microgram . ml-1 of the compound showed malignant transformations 14 weeks after treatment. The treatment with higher concentrations did not accelerate the transformation process.

[Spontaneous transformation and long term culture of foetal mouse glial cells (author's transl)]. / Transformation spontanée et maintien prolongé in vitro de cellules gliales d'embryons de souris.

Six cell lines originated from foetal mouse brain were maintained in vitro for more than two years. The morphology of most of these cultures suggests their glial character. This is corroborated, for some lines, by the presence of specific glial protein (Glial fibrillary acidic protein, or GFAP). Five out of these lines remained not transplantable in animals during all the experiments. However one of them went through a spontaneous transformation at the eleventh population doubling. This culture, after transformation remained glial and when injected in animals, induced GFAP containing tumors.

[Radiological aspects of cancer of the breast in men (author's transl)]. / Aspects radiologiques du cancer du sein chez l'homme.

The radiological signs of malignant mammary tumours in men, which are similar to those observed in women, are reviewed. Several points have to be remembered, however; the common retro-mammillary location, the frequent nodular character, the skin involvement and the usual mamillary retraction, and, finally, the absence of characteristic microcalcifications. The interest of mammography to rate T in the TNM classification is also mentioned.