Comparative Analysis of Three Workload Measurement Methodologies in Surgical Pathology: Conclusions and Implications on Public Health Care and Costing of Pathology Services.

OBJECTIVES: To carry out a comparative analysis between 3 different workload measurement systems in surgical pathology: the Resource-Based Relative Value Scale (RBRVS), the Level 4 Equivalent (L4E), and the Automatable Activity-Based Approach to Complexity Unit Scoring (AABACUS). The RBRVS is one of the most widely used systems in terms of attempting to measure workload, whereas it has been proposed as a means of costing (and thus setting reimbursement rates) of surgical pathology services in Greece, despite being widely criticized for its inaccurate design. METHODS: Surgical pathology workload for 1 representative month at Evaggelismos General Hospital was assessed using both the RBRVS and the 2 newer methods. RESULTS: Pearson correlation showed a high level of correlation (0.902, P < .01) between the L4E and AABACUS but less so between either of those and the RBRVS (0.712 and 0.626, respectively; P < .01). The highest level of discrepancy was observed in the subspecialties of genitourinary, breast, dermatopathology, and gastrointestinal pathology. In addition, total and average working hours as calculated by the RBRVS were significantly lower compared with the other 2 systems. CONCLUSIONS: The RBRVS tends to underestimate actual workload as a result of its inability to take specific workload parameters into account, such as slide count or the need for intradepartmental consultation.

Inhibition of SGK1 enhances mAR-induced apoptosis in MCF-7 breast cancer cells.

Functional membrane androgen receptors (mAR) have previously been described in MCF-7 breast cancer cells. Their stimulation by specific testosterone albumin conjugates (TAC) activate rapidly non-genomic FAK/PI3K/Rac1/Cdc42 signaling, trigger actin reorganization and inhibit cell motility. PI3K stimulates serum and glucocorticoid inducible kinase SGK1, which in turn regulates the function of mAR. In the present study we addressed the role of SGK1 in mAR-induced apoptosis. TAC-stimulated mAR activation elicited apoptosis of MCF-7 cells, an effect significantly potentiated by concomitant incubation of the cells with TAC and the specific SGK1 inhibitors EMD638683 and GSK650394. In line with this, TAC and EMD638683 activated caspase-3. These effects were insensitive to the classical androgen receptor (iAR) antagonist flutamide, pointing to iAR-independent, mAR-induced responses. mAR activation and SGK1 inhibition further considerably augmented the radiation-induced apoptosis of MCF-7 cells. Moreover, TAC- and EMD638683 triggered early actin polymerization in MCF-7 cells. Blocking actin restructuring with cytochalasin B abrogated the TAC- and EMD638683-induced pro-apoptotic responses. Further analysis of the molecular signaling revealed late de-phosphorylation of FAK and Akt. Our results demonstrate that mAR activation triggers pro-apoptotic responses in breast tumor cells, an effect significantly enhanced by SGK1 inhibition, involving actin reorganization and paralleled by down-regulation of FAK/Akt signaling.

Enhanced Orai1 and STIM1 expression as well as store operated Ca2+ entry in therapy resistant ovary carcinoma cells.

Mechanisms underlying therapy resistance of tumor cells include protein kinase Akt. Putative Akt targets include store-operated Ca(2+)-entry (SOCE) accomplished by pore forming ion channel unit Orai1 and its regulator STIM1. We explored whether therapy resistant (A2780cis) differ from therapy sensitive (A2780) ovary carcinoma cells in Akt, Orai1, and STIM1 expression, Ca(2+)-signaling and cell survival following cisplatin (100 µM) treatment. Transcript levels were quantified with RT-PCR, protein abundance with Western blotting, cytosolic Ca(2+)-activity ([Ca(2+)]i) with Fura-2-fluorescence, SOCE from increase of [Ca(2+)]i following Ca(2+)-readdition after Ca(2+)-store depletion, and apoptosis utilizing flow cytometry. Transcript levels of Orai1 and STIM1, protein expression of Orai1, STIM1, and phosphorylated Akt, as well as SOCE were significantly higher in A2780cis than A2780 cells. SOCE was decreased by Akt inhibitor III (SH-6, 10 µM) in A2780cis but not A2780 cells and decreased in both cell lines by Orai1 inhibitor 2-aminoethoxydiphenyl borate (2-ABP, 50 µM). Phosphatidylserine exposure and late apoptosis following cisplatin treatment were significantly lower in A2780cis than A2780 cells, a difference virtually abolished by SH-6 or 2-ABP. In conclusion, Orai1/STIM1 expression and function are increased in therapy resistant ovary carcinoma cells, a property at least in part due to enhanced Akt activity and contributing to therapy resistance in those cells.