Heat shock protein 90 inhibition sensitizes acute myelogenous leukemia cells to cytarabine

Previous studies demonstrated that ataxia telangiectasia mutated- and Rad3-related (ATR) kinase and its downstream target checkpoint kinase 1 (Chk1) facilitate survival of cells treated with nucleoside analogs and other replication inhibitors. Recent results also demonstrated that Chk1 is depleted when cells are treated with heat shock protein 90 (Hsp90) inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG). The present study examined the effects of 17-AAG and its major metabolite, 17-aminogeldanamycin (17-AG), on Chk1 levels and cellular responses to cytarabine in human acute myelogenous leukemia (AML) cell lines and clinical isolates. Cytarabine, at concentrations as low as 30 nM, caused activating phosphorylation of Chk1, loss of the phosphatase Cdc25A, and S-phase slowing. Conversely, treatment with 100 to 300 nM 17-AAG for 24 hours caused Chk1 depletion that was accompanied by diminished cytarabine-induced S-phase accumulation, decreased Cdc25A degradation, and enhanced cytotoxicity as measured by inhibition of colony formation and induction of apoptosis. Additional studies demonstrated that small inhibitory RNA (siRNA) depletion of Chk1 also sensitized cells to cytarabine, whereas disruption of the phosphatidylinositol 3-kinase (PI3k) signaling pathway, which is also blocked by Hsp90 inhibition, did not. Collectively, these results suggest that treatment with 17-AAG might represent a means of reversing checkpoint-mediated cytarabine resistance in AML.     

Computer-mediated interdisciplinary teams: theory and reality

The benefit of experience, tempered with the wisdom of hindsight and 5 years of text-based, asynchronous, computer-mediated, interdisciplinary team communications, provides the energy, insights and data shared in this article. Through the theoretical lens of group dynamics and the epistemology of interdisciplinary teaming, we analyze the interactions of a virtual interdisciplinary team to provide an understanding and appreciation of collaborative interdisciplinary communication in the context of interactive technologies. Whilst interactive technologies may require new patterns of language similar to that of learning a foreign language, what is communicated in the interdisciplinary team process does not change. Most important is the recognition that virtual teams, similar to their face-to-face counterparts, undergo the same challenges of interdisciplinary teaming and group developmental processes of formation: forming, storming, norming, performing, and transforming. After examining these dynamics of communication and collaboration in the context of the virtual team, the article concludes with guidelines facilitating interdisciplinary team computer-mediated communication.     

Infectious keratitis after LASIK

PURPOSE: To report the clinical course, management, and outcomes of culture-proven infectious keratitis in 15 eyes of 13 subjects after LASIK. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Fifteen eyes of 13 subjects who underwent LASIK and developed culture-positive keratitis. INTERVENTION: Infectious keratitis was encountered in the operative eyes between 1 day and 450 days. Cultures were obtained, and topical antibiotic therapy was administered in all cases. Some cases required flap lifting, irrigation, and soaking of the bed with antibiotics, flap amputation, or further surgical intervention. MAIN OUTCOME MEASURES: Time periods from onset to diagnosis, from clinical diagnosis to clinical resolution, final acuities, microbiologic profiles, and medical and surgical interventions were reviewed. RESULTS: Onset of symptoms of infection varied, depending on the infectious organism. Bacterial organisms tended to present earlier, whereas mycobacterial and fungal organisms had a later mean onset of presentation. Furthermore, the atypical organisms such as mycobacteria, fungus, and acanthamoeba also had a more delayed diagnosis, resulting in a prolonged disease course. CONCLUSIONS: Infectious keratitis after LASIK is a potentially vision-threatening complication. Onset of symptoms varies depending on causative agents. Furthermore, atypical organisms in the interface or beneath the flap can pose both diagnostic and therapeutic dilemmas. Location in the interface can make it more difficult to culture the organisms and prevent adequate penetration of topical antibiotics.     

Corneal flap complications in refractive surgery: Part 1: development of an experimental animal model

PURPOSE: To report the outcome, learning curve, and complication rates of an experimental animal model for corneal flaps in refractive surgery. SETTING: Magill Research Center for Vision Correction, Storm Eye Institute, Charleston, South Carolina, USA. METHODS: Corneal flaps with a nasal or a temporal hinge were created in 190 eyes of 95 Dutch Belted rabbits using the Automated Corneal Shaper microkeratome (Bausch & Lomb Surgical). Diffuse lamellar keratitis (DLK) was induced by inoculating the corneal interfaces with 1 of 7 substances. Postoperatively, the eyes were examined with a slitlamp. Special emphasis was placed on corneal flap complications and the relationship between slipped flaps and hinge position and/or inoculation agent. RESULTS: A good corneal flap was achieved in 174 eyes (92%). The eyes with a nasal hinge had a lower incidence of slipped flaps (14%) than eyes with a temporal hinge (37%) (P =.02). CONCLUSION: With the animal model described, corneal flaps were created in a precise and reproducible way in more than 90% of eyes. Nasal hinged flaps showed less postoperative displacements than temporal hinged flaps and are adequate for further study.     

How should the effectiveness of problem-based learning in occupational therapy education be examined?

Our aim in discussing the issue of educational outcomes of PBL has been to provoke discussion among occupational therapy educators and researchers. Future collaboration and research should involve occupational therapy educational programs that use PBL and those who have yet to access this teaching-learning method. Multisite studies would allow the evaluation of different teaching methods of clinical reasoning and their respective efficacies. Germane instruments that are easy to use and that effectively measure clinical reasoning in occupational therapy must be developed to supplement the qualitative research that is being undertaken. Such research will support and validate the use of resources and constructively contribute to accountability. PBL may be valuable for promoting adaptive learning in students, which will help them and the profession stay informed and proactive and ensure best practice in a rapidly changing health care environment. This article is a call for educators to collaborate in educational outcome research studies. The traditional methods of evaluation, such as pass rates on national certification examinations, may not demonstrate the multidimensional learning and cognitive growth acquired in PBL; therefore, there is a need to generate validating research about the efficacy of PBL in occupational therapy education. If we are to undertake the challenging task of measuring the development of clinical reasoning, research will require creativity and reflection, hallmarks of the PBL process.     

Interaction of high molecular weight kininogen, factor XII, and fibrinogen in plasma at interfaces

Using ellipsometry, anodized tantalum interference color, and Coomassie blue staining in conjunction with immunologic identification of proteins adsorbed at interfaces, we have previously found that fibrinogen is the main constituent deposited by plasma onto many man- made surfaces. However, the fibrinogen deposited from normal plasma onto glass and similar wettable materials is rapidly modified during contact activation until it can no longer be identified antigenically. In earlier publications, we have called this modification of the fibrinogen layer "conversion," to indicate a process of unknown nature. Conversion of adsorbed fibrinogen by the plasma was not accompanied by marked change in film thickness, so that we presumed that this fibrinogen was not covered but replaced by other protein. Conversion is now showen to be markedly delayed in plasma lacking high molecular weight kininogen, slightly delayed in plasma lacking factor XII, and normal in plasma that lack factor XI or prekallikrein. We conclude that intact plasma will quickly replace the fibrinogen it has deposited on glass-like surfaces by high molecular weight kininogen and, to a smaller extent, by factor XII. Platelets adhere preferentially to fibrinogen-coated surfaces; human platelets adhere to hydrophobic nonactivating surfaces, since on these, adsorbed firbinogen is not exchanged by the plasma. The adsorbed fibrinogen will be replaced on glass-like surfaces during surface activation of clotting, and platelets failing to find fibrinogen will not adhere.     

Isolation and morphologic characterization of bile duct epithelial cells from normal rat liver

To study directly the functions of the cells that line the bile ducts inside the liver, we developed a new technique for isolating intrahepatic bile duct epithelial cells (IBDECs) from normal rat liver. Parenchymal and nonparenchymal cells were separated from whole liver by enzymatic digestion and mechanical disruption; subpopulations of individual nonparenchymal cells then were isolated by serial counter-flow elutriation, isopycnic centrifugation, and immunoaffinity separation with a specific monoclonal antibody against an antigen on the plasma membrane of IBDECs. Using this approach, we isolated 1.2 +/- 0.2 x 10(6) (mean +/- SE) viable (greater than 95% trypan blue exclusion) cells, greater than 95% of which were identified as IBDECs by morphologic appearance and specific cytochemical markers. The IBDECs averaged 7.4 +/- 0.16 microM in diameter and retained their in situ appearance, including morphologic polarity. They appeared as single cells or as cell doublets attached by tight junctions that excluded ruthenium red. Microvilli were abundant and were restricted to the apical (i.e., luminal) domain of the plasma membrane. Coated pits were observed on both apical and basolateral cell surfaces. Internally, IBDECs contained a well-developed system of organelles, including mitochondria, Golgi, and discrete types of vesicles, such as coated vesicles, multivesicular bodies, and lysosomes. These results indicate that a highly purified suspension of viable, morphologically intact, and polar IBDECs can be prepared from normal rat liver using a novel approach that separates liver cells on the basis of size, density, and specific membrane components. The availability of such a model will allow experimental studies to be performed directly on IBDECs, an approach that has not previously been possible.