Anticancer effects of resveratrol in canine hemangiosarcoma cell lines.

Hemangiosarcoma (HSA) is a highly malignant tumour with aggressive biological behaviour. HSAs are more common in dogs than other domestic animals. The median survival time of dogs with HSA remains short, even with chemotherapy and surgery. Therefore, there is a critical need to improve the adjuvant chemotherapeutic regimens to improve clinical outcomes in dogs with HSA. Resveratrol has been shown to possess strong anti-proliferative and/or pro-apoptotic properties in human cancer cell lines. Nevertheless, the potential anticancer effects of resveratrol have not been reported in canine HSAs. The objective of this study is to determine the growth inhibitory effects of resveratrol in HSA cells when used alone or in combination with doxorubicin, a commonly used chemotherapeutic agent. Frog and DD-1 canine HSA cell lines were treated with varying concentrations of resveratrol with and without doxorubicin. Cell viability was measured by the MTT assay. The expression of apoptotic proteins, activation of p38 mitogen-activated protein kinase (MAPK), AMP-activated protein kinase (AMPK) and extracellular signal-regulated kinase 1/2 (ERK1/2) were assessed by western blotting. Similar to human cancer cell lines, resveratrol markedly inhibited the growth and induced apoptosis in both HSA cell lines. Mechanistically, resveratrol activated p38 MAPK, but did not affect the AMPK or the ERK1/2 pathways. Additional experiments showed that resveratrol augmented the growth-inhibitory and apoptotic effects of doxorubicin in both HSA cell lines. These findings suggest that resveratrol has pro-apoptotic effects in canine HSA cells; therefore, its use as a potential adjunct therapy in canine HSA patients warrants further investigation.

Synthesis and pharmacological evaluation of dimeric muscarinic acetylcholine receptor agonists.

Two dimeric analogs of the muscarinic acetylcholine receptor (mAChR) agonist phenylpropargyloxy-1,2,5-thiadiazole-quinuclidine (NNC 11-1314) were synthesized and pharmacologically evaluated. In radioligand binding assays on Chinese hamster ovary (CHO) cell membranes expressing the individual human M(1) to M(5) mAChR subtypes, both dimers [(3S)-1,4-bis-(3-[(3-azabicyclo[2.2.2]octanyl)-1,2,5-thiadiazol-4-yloxy]-1-propyn-1-yl)benzene,2-L-(+)-tartrate (NNC 11-1607) and (3S)-1,3-bis-(3-[(3-azabicyclo[2.2.2]octanyl)-1,2,5-thiadiazol-4-yloxy]-1-propyn-1-yl)benzene,2-L-(+)-tartrate (NNC 11-1585)] exhibited higher binding affinities than the monomeric NNC 11-1314. Only NNC 11-1585, however, displayed significant selectivity for the M(1) and M(2) mAChRs relative to the other subtypes. Although binding studies in rat brain homogenates supported the selectivity profile of NNC 11-1585 observed in the CHO membranes, rat heart membrane experiments revealed complex binding behavior for all three agonists that most likely reflected differences in species and host cell environment between the heart and CHO cells. Subsequent functional assays with phosphatidylinositol hydrolysis revealed that all three novel ligands were partial agonists relative to the full agonist oxotremorine-M at the CHO M(1), M(3), and M(5) mAChRs, with NNC 11-1607 displaying the highest functional selectivity. In the CHO M(2) and M(4) mAChR cells, agonist-mediated effects on forskolin-stimulated cAMP accumulation were characterized by bell-shaped concentration-response curves, with the exceptions of NNC 11-1607, which had no discernible effects at the M(2) mAChR, and NNC 11-1585, which could only inhibit cAMP accumulation at the M(4) mAChR. Thus, we identified NNC 11-1607 as a novel functionally selective M(1)/M(4) mAChR agonist. Our data suggest that dimerization of mAChR agonists is a viable approach in designing more potent and functionally selective agonists, as well as in providing novel tools with which to probe the nature of agonism at these receptors.

Transducer abstraction: a novel approach to the detection of partial agonist efficacy in radioligand binding studies.

The properties of the ternary complex model (TCM) of drug action at G protein-coupled receptors (GPCRs) were examined, using theoretical computer simulations, with regard to the predicted effects of the presence of a fixed concentration of one agonist on the competition binding profile of another. Subsequently, the binding properties of the full muscarinic acetylcholine receptor (mAChR) agonists acetylcholine (ACh) and carbachol (CCh), and the partial agonists pilocarpine and McN-A-343, were investigated in competition experiments against [(3)H]N-methylscopolamine using homogenate preparations from Chinese hamster ovary cells, stably expressing the human M(1) or M(2) mAChR. At the M(2) mAChR, all agonists displayed biphasic binding curves and were readily modulated by the non-hydrolyzable GTP analogue, Gpp(NH)p, in accordance with previously established experimental observations. In contrast, agonist binding at the M(1) mAChR showed no significant change in the presence of Gpp(NH)p, even in the case of a full agonist. This phenomenon precludes using the "GTP-shift" to assess agonist efficacy at the M(1) mAChR. When the ACh competition curves were reconstructed in the presence of graded concentrations of either a full or a partial agonist, a significant redistribution of the fraction of the high-affinity state recognized by ACh was observed. However, when the procedure was repeated using the antagonist, atropine, no significant effect on the fraction of either the high or low affinity ACh binding components at the mAChR was observed. Taken together, these results indicate that changes in the profile of full agonist binding isotherms, when constructed in the presence of a partial agonist, may be more sensitive indicators of partial agonist efficacy than regular assays that directly measure partial agonist binding.

Effects of speech therapy and pharmacologic and surgical treatments on voice and speech in Parkinson's disease: a review of the literature.

The purpose of this review was to examine the different treatment approaches for persons with Parkinson's Disease (PD) and to examine the effects of these treatments on speech. Treatment methods reviewed include speech therapy, pharmacological, and surgical. Research from the 1950s through the 1970s had not demonstrated significant improvements following speech therapy. Recent research has shown that speech therapy (when persons with PD are optimally medicated) has proven to be the most efficacious therapeutic method for improving voice and speech function. Pharmacological methods of treatment in isolation do not appear to significantly improve voice and speech function in PD across research studies. Surgical treatment methods including pallidotomy and deep brain stimulation may be significant treatment options which improve voice and speech function in some persons with PD. Possible explanations for the differential responses to treatment are discussed. Future studies should investigate the effects of combined treatment approaches. Perhaps the combination of pharmacological, surgical and speech treatment will prove superior to treatments combining pharmacological and surgical or pharmacological and speech therapy in improving the communication abilities of persons with PD.

Regulation of acetylcholine binding by ATP at the muscarinic M(1) receptor in intact CHO cells.

ATP may have a modulatory effect on cholinergic transmission, as it is known that ATP is released as a co-transmitter with acetylcholine from nerve terminals. The ability of ATP to influence the binding of acetylcholine to the M(1) muscarinic acetylcholine receptor expressed in intact CHO cells was investigated. In competition binding experiments, acetylcholine completely inhibited the binding of [3H]N-methylscopolamine, but yielded a shallow competition isotherm that was best described in terms of two affinity states. When these experiments were repeated in the presence of 1 mM ATP, the acetylcholine competition curve was better described in terms of a single, low-affinity state with a Hill slope not significantly different from unity. This modulatory effect of ATP was completely reversed by the addition of the P(2) purinoceptor antagonist, suramin, to the assay medium. When the competition between the muscarinic receptor antagonist, atropine, and [3H]N-methylscopolamine was investigated, however, ATP was unable to modulate the binding of atropine, which was consistent with a one-site binding model in each instance. In contrast to the intact cell studies, ATP did not affect either affinity state of acetylcholine binding when studied in homogenate preparations. The results of the present study indicate that ATP, acting via endogenously expressed purinoceptors, is able to influence agonist binding to the M(1) muscarinic acetylcholine receptor via a cross-talk that requires the functional integrity of intact CHO cells.

The necessity of mission integration. A system develops processes to weave values into the life of the organization.

Essential to the future vitality and viability of a mission-driven organization is the integration of the mission into the organization's programs, policies, practices, and accountability. Holy Cross Health System (HCHS), South Bend, IN, launched an intensive educational effort with managers, staff members, and trustees to reinforce the basic belief that mission permeates all departments. Using the mission statements principles of fidelity, excellence, empowerment, and stewardship, HCHS leaders initiated a systemwide mission assessment and development effort. During assessment, each facilities' ad hoc team addressed and responded to the organization's mission standards on the basis of availability of personnel, size, facility, and particular circumstances. The assessment process called for interdisciplinary, institutional review teams to explore all aspects of mission activity. This process enabled HCHS to launch a systemwide educational effort about the importance and necessity of mission integration. HCHS then used the mission statement elements fidelity, excellence, empowerment, and stewardship to define new relationships of accountability.

Sponsorship in evolution.

Sponsorship appears to be evolving from an original model in which the sponsoring religious institute related to its facilities in a manner resembling a family business, to a model of sponsorship akin to a franchise, to a ministerial partnership. Factors leading to this evolution include tremendous changes within the religious institute itself, including decreases in the number of members and financial stability. Changes within healthcare itself--such as greater competition and declining revenues-have forced hospitals to diversify. One result of these developments has been a radical change in the "rules" of the game. Historically independent entities--hospitals, sponsors, physicians--now have to value interdependence and mutuality. In the family-run model the family (sponsor) had special privileges, as though they "owned" the business. When the number of family members dropped below that necessary to govern, administer, and staff the institute's facilities, they began to move away from the family model to the franchise model, which has more open communication, greater input to decision making by non-family members, and a shift in the family's attention from actual operations to oversight and accountability. Eventually, the franchise model began to give way to the ministerial partnership, characterized by mutuality. Both family and others have roles not only in carrying out the mission, but in actually shaping and forming it.

The call to collaboration. Pursuing and completing a merger requires planning and perseverance.

In 1987 Mercy Health Care System (MCHS) of Scranton, PA, and Mercy Health System (MHS) of Cincinnati took the first steps toward a merger. Leaders of the Scranton province, who had for some time been seeking to establish a relationship with another Mercy-sponsored health system, had approached MHS with the proposal. In late summer 1988 MHS began a thorough analysis of the facilities of the Pennsylvania system, assessing their mission, margin, and market strength; plant and property conditions; and human resources. At the same time the two provincial teams and CEOs from facilities in both systems, along with an outside consultant, began to negotiate the proposed merger's sponsorship and governance structure. Careful avoidance of "we-they" thinking was the hallmark of the entire negotiating process. Having agreed that they wanted a cosponsored system, the negotiating teams struggled to determine the degree of representation the Scranton province (five facilities) and the Cincinnati province (18 facilities) would have. The teams also had to iron out differences of opinion regarding the viability of one facility. Negotiators also worked out a decision-making procedure ensuring appropriate input from the Scranton member of the corporation. Help was sought to ensure that the merger agreement was in accordance with both canon and civil law. A sophisticated, step-by-step implementation process helped bring MHCS facilities on line as quickly as possible with minimal disruption. A series of celebrations commemorated the merger's completion.