Estrogen receptor domains E and F: role in dimerization and interaction with coactivator RIP-140

We have used the yeast two-hybrid system to localize the ligand-dependent dimerization domain of the estrogen receptor-alpha (ER) to region E in vivo. In this system, the cDNAs corresponding to the A-D, E, E/F, A-E (deltaF), and full-length (wtER) domains of the human ER were each cloned into the yeast two-hybrid vectors GAL4 DB and GAL4 TA and expressed in different combinations in yeast harboring a GAL1-lacZ reporter. The reporter was used as a relative measure of the interaction between the ER domains, through reconstitution of GAL4 activity. We found that the interaction of E or E/F domains of the ER with full-length ER is estradiol dependent and estrogen responsive element independent, as measured by the reconstitution of GAL4 activity from GAL4-E domain-containing fusion protein interactions. In the presence of F domain, this activity is reduced 10-fold. The results suggest that sequences in the F domain are inhibitory to the dimerization signal that is present in the E region. We propose that the full-length ER contains intrinsic dimerization restraints contributed by regions outside domain E that are released upon binding hormone agonist. In addition, we have demonstrated that coactivator RIP140 is able to interact with the ER in vivo at the E domain of the receptor in the presence of estrogen. Yeast two-hybrid analysis shows that RIP140 does not homodimerize in the presence or absence of estrogens. We present evidence showing that the ER has the inherent ability to interact with RIP140 in the presence of antiestrogens, but sequences inherent in the ER itself that are present outside of the E domain compromise this ability.     

Billing third party payers for pharmaceutical care services

OBJECTIVE: To describe the steps pharmacists must complete when seeking compensation from third party payers for pharmaceutical care services. DATA SOURCES: Government publications; professional publications, including manuals and newsletters; authors' personal experience. DATA SYNTHESIS: Pharmacists in increasing numbers are meeting with success in getting reimbursed by third party payers for patient care activities. However, many pharmacists remain reluctant to seek compensation because they do not understand the steps involved. Preparatory steps include obtaining a provider/supplier number, procuring appropriate claim forms, developing data collection and documentation systems, establishing professional fees, creating a marketing plan, and developing an accounting system. To bill for specific patient care services, pharmacists need to collect the patient's insurance information, obtain a statement of medical necessity from the patient's physician, complete the appropriate claim form accurately, and submit the claim with supporting documentation to the insurer. Although many claims from pharmacists are rejected initially, pharmacists who work with third party payers to understand the reasons for denial of payment often receive compensation when claims are resubmitted. CONCLUSION: Pharmacists who follow these guidelines for billing third party payers for pharmaceutical care services should notice an increase in the number of paid claims.     

The binding of 3,6-disubstituted bile salts to human serum albumin induces conformational change on the molecule of this protein

The binding of 3,6-hydroxy and keto disubstituted bile salts to human serum albumin was studied using differential scanning calorimetry, fluorescence spectroscopy and circular dichroism. The bile salts assayed did not produce any modification in the shape of the albumin thermogram, its thermal unfolding process in their presence being reversible; however, an increase in the enthalpy of unfolding and in the Tm was observed in the presence of 3,6-diketo and 3-hydroxy-6-keto bile salts. These two derivatives induced a negative circular dichroism spectrum of the protein around 280-290 nm, quenched the native fluorescence of the buried tryptophan of albumin and induced energy transfer between 1 aniline-8-naphthalene sulfonate and the buried tryptophan 214 of albumin. The presence of a keto group at C6 in the steroid ring of the bile salts plays an important role in producing slight movement of the albumin domains, increasing the distance between domains I and II.     

Unresponsiveness to cannabinoids and reduced addictive effects of opiates in CB1 receptor knockout mice

The function of the central cannabinoid receptor (CB1) was investigated by invalidating its gene. Mutant mice did not respond to cannabinoid drugs, demonstrating the exclusive role of the CB1 receptor in mediating analgesia, reinforcement, hypothermia, hypolocomotion, and hypotension. The acute effects of opiates were unaffected, but the reinforcing properties of morphine and the severity of the withdrawal syndrome were strongly reduced. These observations suggest that the CB1 receptor is involved in the motivational properties of opiates and in the development of physical dependence and extend the concept of an interconnected role of CB1 and opiate receptors in the brain areas mediating addictive behavior.     

Noise sources across a normal-to-superconducting boundary in bulkYBa2Cu3O7-x

Experiments in which a normal-to-superconducting (NS) boundary is created in an originally superconducting bar of bulk YBa₂Cu ₃O_7-x is described. The first noise measurements associated with such a boundary are presented. A high-density (5.20 g-cm ^-3) sample (sample A) and a low density (4.23 g-cm^-3 ) sample (sample B) have been investigated. Common to both samples is 1/f² noise observed in the frequency region between about 15 and 50 Hz that can be associated with the NS boundary itself. Sample A also exhibits conventional 1/f noise as expected for the normal part of the boundary, below about 15 Hz. In contrast, sample B shows 1/f^1.5 noise in this region; since the sample contains considerably fewer small grains than sample A, any 1/f noise is presumably masked in B. It is suggested that the 1/f² noise might be associated with flux-flow noise     

Surgical management of meningiomas originating in Meckel's cave

OBJECTIVE: To define the difference of meningiomas that originate in the area of Meckel's cave (MC) (primary MC meningiomas) in regard to the different surgical approaches and postoperative results. METHODS: A retrospective analysis of all meningiomas involving the cranial base displayed 21 cases of meningiomas originating in MC (primary MC meningiomas). These cases were classified according to the tumor extension in four different types: Type I, tumors mainly confined to MC; Type II, MC meningiomas with extension into the middle fossa; Type III, MC meningiomas with extension into the posterior fossa; and Type IV, MC meningiomas with extension into both middle and posterior fossae. RESULTS: Trigeminal neuralgia resolved in all cases in this series, despite tumor type. Trigeminal hypesthesia showed postoperative improvement only in Type III MC meningiomas. In Types I and III, total removal without further morbidity was frequently achieved. Cavernous sinus infiltration, especially in Types II and IV, limited (in some cases) the extent of tumor extirpation. CONCLUSION: Types I, II, and III MC meningiomas have a good prognosis. In most cases, very good outcomes are achieved. Radical tumor removal can usually be achieved without further morbidity and with postoperative improvement of the preexisting symptoms, especially in Types I and III MC meningiomas. On the contrary, Type IV MC meningiomas are usually only subtotally resected. Surgery in such cases may carry a high risk of additional morbidity, especially with regard to the IIIrd, IVth, and VIth cranial nerves. The postoperative outcome regarding facial pain in cases of all tumor types is usually very good. Trigeminal hypesthesia may persist after tumor removal in the majority of cases.