A dental health education program for Boston's Native American community

PLEASE NOTE: This work is protected by copyright. Downloading is restricted to the BU community: please click Download and log in with a valid BU account to access. If you are the author of this work and would like to make it publicly available, please contact [email protected] program is a joint effort of the Boston Indian Council and the Goldman School of Gradiate Dentistry.Thesis (M.S.)--Boston University, Henry M. Goldman School of Graduate Dentistry, 1977 (Dental Public Health)Bibliography: leaves 16-17.A three phase dental program was initiated during 1977 to help meet the needs of Boston’s Native American community. A series of five dental health education workshops for adults in the community was designed to accomplish the following goals: 1.to improve the overall attitude toward dentistry among the Native Americans in the community 2.to improve the oral hygiene status among workshop participants 3.to reduce the fear of seeking dental treatment as a barrier to utilization of services Phase one was a series of five weekly dental health education workshops conducted for adults in the Indian community during the months of February and March, 1977. As a result of the workshops, adults in the Indian community are now seeking preventive dental treatment at Boston University School of Graduate Dentistry (BUSGD) on a regular basis. Phase two was the design of a weekly fluoride mouth rinse program for forty children involved in an after school class at the Boston Indian Council (B.I.C.). Staff at the B.I.C. have been trained to administer the program. The arrangement of transportation to a neighborhood health center for Indian children to receive dental treatment was the third phase of the program. This program is still in the planning process as of May, 1977. Results of the three phase program indicated that the main barriers preventing access to dental care in this community are 1ack of transportation and inadequate funding. Therefore, future programs for this community should emphasize the elimination of these barriers. Through the efforts of concerned individuals, the health status of Boston's Indians will gradually improve

Analysis of the Bile Salt Export Pump (<i>ABCB11</i>) Interactome Employing Complementary Approaches

<div><p>The bile salt export pump (BSEP, <i>ABCB11</i>) plays an essential role in the formation of bile. In hepatocytes, BSEP is localized within the apical (canalicular) membrane and a deficiency of canalicular BSEP function is associated with severe forms of cholestasis. Regulation of correct trafficking to the canalicular membrane and of activity is essential to ensure BSEP functionality and thus normal bile flow. However, little is known about the identity of interaction partners regulating function and localization of BSEP. In our study, interaction partners of BSEP were identified in a complementary approach: Firstly, BSEP interaction partners were co-immunoprecipitated from human liver samples and identified by mass spectrometry (MS). Secondly, a membrane yeast two-hybrid (MYTH) assay was used to determine protein interaction partners using a human liver cDNA library. A selection of interaction partners identified both by MYTH and MS were verified by <i>in vitro</i> interaction studies using purified proteins. By these complementary approaches, a set of ten novel BSEP interaction partners was identified. With the exception of radixin, all other interaction partners were integral or membrane-associated proteins including proteins of the early secretory pathway and the bile acyl-CoA synthetase, the second to last, ER-associated enzyme of bile salt synthesis.</p></div

co-IP / MS/MS identifies BSEP interaction partners.

<p><b>(A)</b> Immunoprecipitation of BSEP coupled to complex mass spectrometry reveals new interaction partners of BSEP in human liver. Crude canalicular membrane preparations were solubilized in either digitonin or Triton X-100, and immunoprecipitated samples were subjected to MS/MS. For each detergent, protein frequencies from two co-IPs with BSEP antibody are plotted against the respective negative control with naïve mouse IgG. Interaction partners of interest are labeled. For the sake of clarity, proteins found exclusively in either the BSEP or control co-IP are depicted with an MS-score of one instead of zero on the other axis. (<b>B)</b> Immunoblot analysis of co-IPs shown inA.</p

Proteins identified as interaction partners of BSEP in MYTH and co-IP / MS/MS screens.

<p>Proteins identified as interaction partners of BSEP in MYTH and co-IP / MS/MS screens.</p

BSEP interacts with radixin and the bile acyl-CoA synthetase.

<p><b>(A)</b> MYTH bait dependency test of radixin_1-318 and BACS_640-690 against BSEP or a non-interacting control bait. (<b>B)</b> Pull-down of BSEP with radixin_1-318, BACS_77-690 and AP-2 μ1. Protein interaction partners were immobilized as bait on Strep-Tactin Sepharose and purified BSEP was added. BSEP and the interaction partners were detected by immunoblot analysis with monoclonal antibodies against BSEP or the Strep-tag, respectively. Strep-Tactin Sepharose without bait protein served as negative control. (<b>C)</b> Tag-less BSEP can be pulled down with radixin_1-318, but not with full-length, non-activated radixin. Protein interaction partners were immobilized on Strep-Tactin Sepharose. Purified, tag-less BSEP was added to the beads and the complexes were eluted after washing. BSEP and the interacting proteins were detected by immunoblot analysis with monoclonal antibodies against BSEP or the Strep-tag, respectively.</p

Schematic diagram of the membrane yeast two-hybrid system (MYTH) to screen for interaction partners of the ABC-transporter BSEP.

<p>The MYTH system is based on a split-ubiquitin approach. The bait, BSEP, is fused to the C-terminus of ubiquitin (Cub) and a transcription factor (LexA-VP16). The preys are soluble or membrane-associated liver proteins, introduced via a cDNA library. They are fused to the N-terminus of ubiquitin (NubG). Upon interaction of BSEP and the liver protein the ubiquitin moieties, which have a low affinity for each other due to a mutation in the N-terminus, come into close proximity. The reassembled ubiquitin is recognized by endogenous ubiquitin specific proteases (UBP). The transcription factor is cleaved off and activates the reporter genes (<i>HIS3</i>, <i>ADE2</i>, <i>lacZ</i>).</p

Bait dependency test with BSEP interaction partners in the early secretory pathway.

<p>To confirm the interaction with prey proteins from the initial MYTH screen, reporter gene activation was tested individually for each prey against the bait, BSEP, or a non-interacting control bait, large T antigen. The figure shows one representative result for the positively tested preys, which are denoted by their respective gene name.</p

Functional control assay for the membrane yeast two-hybrid with BSEP and self-activation control.

<p>The yeast strain NMY51 was transformed with the BSEP bait construct and control plasmids coding either for a nonsense peptide with the NubI (wild-type ubiquitin) or the NubG (mutated ubiquitin) -tag. Due to its affinity to its C-terminal half the NubI moiety activates the system regardless of bait interaction. Both controls show equal transformation efficiency on selective media (SD-LW) for the plasmids (upper panels). The positive control shows yeast growth on selective medium (SD-LWH) due to the affinity of the wild-type ubiquitin moieties. This confirms expression of the BSEP fusion protein. In contrast, the negative control shows no reporter gene activation based on unspecific interaction of BSEP with the NubG-nonsense peptide. Additionally, no self-activation of the MYTH system with BSEP was detected in a library-scale transformation of the empty pPR3-N library vector.</p