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Identification of cytoskeletal [14C]carboplatin-binding proteins reveals reduced expression and disorganization of actin and filamin in cisplatin-resistant cell lines.

Cisplatin resistant (CP-r) cells often show decreased uptake of cisplatin in association with reduced cell surface proteins and decreased endocytosis. In this report, two major [14C]carboplatin-binding proteins were identified as filamin and actin by photoaffinity labeling and mass spectrometry. Decreased expression of these two proteins was found in two different human CP-r cell lines (KB-CP20 and 7404-CP20), in comparison with their parental cell lines (KB-3-1 and BEL-7404), respectively. Disorganization of beta-actin and filamin 250 and 90 was also detected in these CP-r cells by confocal microscopy. Transfection of a wild-type actin-enhanced green fluorescent protein (EGFP) expression vector into 7404-CP20 cells resulted in a nonfilamentous actin-EGFP distribution compared with a normal distribution in the cisplatin-sensitive BEL-7404 cells, suggesting that cytoskeletal organization is disturbed in the CP-r cells. The identification of actin and filamin as [14C]carboplatin-binding proteins and decreased expression and disorganization of several cytoskeletal proteins in CP-r cells provide a molecular and cellular basis for the known defect in endocytosis in these cells.

Actins↗

Use of an adaptable cell culture kit for performing lymphocyte and monocyte cell cultures in microgravity.

The results of experiments performed in recent years on board facilities such as the Space Shuttle/Spacelab have demonstrated that many cell systems, ranging from simple bacteria to mammalian cells, are sensitive to the microgravity environment, suggesting gravity affects fundamental cellular processes. However, performing well-controlled experiments aboard spacecraft offers unique challenges to the cell biologist. Although systems such as the European 'Biorack' provide generic experiment facilities including an incubator, on-board 1-g reference centrifuge, and contained area for manipulations, the experimenter must still establish a system for performing cell culture experiments that is compatible with the constraints of spaceflight. Two different cell culture kits developed by the French Space Agency, CNES, were recently used to perform a series of experiments during four flights of the 'Biorack' facility aboard the Space Shuttle. The first unit, Generic Cell Activation Kit 1 (GCAK-1), contains six separate culture units per cassette, each consisting of a culture chamber, activator chamber, filtration system (permitting separation of cells from supernatant in-flight), injection port, and supernatant collection chamber. The second unit (GCAK-2) also contains six separate culture units, including a culture, activator, and fixation chambers. Both hardware units permit relatively complex cell culture manipulations without extensive use of spacecraft resources (crew time, volume, mass, power), or the need for excessive safety measures. Possible operations include stimulation of cultures with activators, separation of cells from supernatant, fixation/lysis, manipulation of radiolabelled reagents, and medium exchange. Investigations performed aboard the Space Shuttle in six different experiments used Jurkat, purified T-cells or U937 cells, the results of which are reported separately. We report here the behaviour of Jurkat and U937 cells in the GCAK hardware in ground-based investigations simulating the conditions expected in the flight experiment. Several parameters including cell concentration, time between cell loading and activation, and storage temperature on cell survival were examined to characterise cell response and optimise the experiments to be flown aboard the Space Shuttle. Results indicate that the objectives of the experiments could be met with delays up to 5 days between cell loading into the hardware and initial in flight experiment activation, without the need for medium exchange. Experiment hardware of this kind, which is adaptable to a wide range of cell types and can be easily interfaced to different spacecraft facilities, offers the possibility for a wide range of experimenters successfully and easily to utilise future flight opportunities.

Biological Transport↗

Mothers' parenting cognitions in cultures of origin, acculturating cultures, and cultures of destination.

Japanese and South American immigrant mothers' parenting cognitions (attributions and self-perceptions) were compared with mothers from their country of origin (Japan and Argentina, respectively) and European American mothers in the United States. Participants were 231 mothers of 20-month-old children. Generally, South American immigrant mothers' parenting cognitions more closely resembled those of mothers in the United States, whereas Japanese immigrant mothers' cognitions tended to be similar to those of Japanese mothers or intermediate between Japanese and U.S. mothers. This study provides insight into the nature of parenting cognitions generally and those of immigrant mothers specifically and therefore the parenting climate in which immigrant children are reared.

Acculturation↗

Case vignette: child abuse or acceptable cultural norms -- child psychiatrist's response; ethical issues in culturally relevant interventions; cultural contexts.

Three families from the nation of the Gambia have become socially acquainted in the greater Metropolis area where they have relocated from their native country. All three families have preadolescent daughters. None of the girls has as yet undergone the ritual "female circumcision" commonly practiced in their native country. Five of the six parents think it would be a good idea to hold such a ritual ceremony in Metropolis next month. The sixth parent is uneasy about the procedure and wonders if it is appropriate anyway. The parent comes to talk with you, a physician or mental health professional, about this. The parent describes concerns about the procedure and the practice in general, but notes that some groups in the United States do "similar things," referring to the ritual circumcision of Jewish boys at 8 days of age. The parent adds that if the procedure is not possible in the United States, it could be done in the Gambia when the family returns there on holiday. Discussants were asked for their opinions regarding advice to the parent's ethical obligations (especially in the case of mandated reporting of suspected child abuse).

Africa↗

Enhancement of liver-specific functions of primary rat hepatocytes co-cultured with bone marrow cells on tissue culture-treated polystyrene surfaces.

Primary rat hepatocytes were co-cultured with bone marrow cells (BMCs) in a tissue culture-treated culture well (TCW) or a collagen-coated culture well (CCW). Although a medium containing serum was used, the co-cultured cells formed spheroids on the TCW within 2 days, and ammonia metabolism and albumin secretion activities were well maintained for 3 weeks. The co-cultured cells formed a monolayer on the CCW within 2 days, and liver functions were maintained for 3 weeks. The ammonia metabolism activities and albumin secretion activities of co-cultured cells on the CCWs were slightly inferior to those on the TCWs. This co-culture system of hepatocytes and BMCs is an appropriate culture system for the expression and maintenance of liver-specific functions in vitro, and is expected to be applicable to bioartificial liver systems, regenerative medicine, and liver function simulators.

Albumins↗