Building Capacity for Behavioral Health Services and Clinical Research in a Rural Primary Care Clinic: A Case Study.

Integrating sustainable, evidence-based, and collaborative depression screening and follow-up treatment into primary care clinics is a significant challenge in health care. In this article a case study approach is used to describe the process of building capacity for a depression screening program in a rural federally qualified health center (FQHC). A conceptual framework addressing the clinical, operational, and financial perspectives of a primary care setting is applied restrospectively to identify 1) the barriers and facilitating factors associated with integrating a depression screening program into standard practice and 2) how the program was leveraged to conduct clinical research to improve self-management in patients with diabetes and elevated depressive symptoms.

Integration of the cytogenetic and physical maps of chicken chromosome 17.

The chicken genome, like those of most avian species, contains numerous microchromosomes that cannot be distinguished by size alone. Unique properties attributed to the microchromosomes include high GC content and gene density, and an enhanced recombination rate. Previously, microchromosome GGA 17 was shown to align with the consensus genetic linkage group E41W17, and bacterial artificial chromosome (BAC) clones containing E41W17 markers were isolated and assigned on the physical BAC map as well as the recently assembled draft chicken genome sequence. For this study, these same BACS were utilized as probes for fluorescence in-situ hybridization (FISH) to develop the GGA 17 cytogenetic map. Here we detail the chromosome order of ten BAC DNAs, thereby deriving a cytogenetic map of GGA 17 that is simultaneously integrated with both the linkage map and genome sequence. The location of the FISH probes together with the morphological appearance of the chromosome suggested that GGA 17 is an acrocentric chromosome whose cytogenetic map orientation is reversed from that currently indicated by the linkage map and draft genome sequence. The reversed orientation and the centromere location of GGA 17 were confirmed experimentally by dual-colour FISH hybridization using terminal BACs and the centromere-specific CNM oligonucleotide as probes. An advantage of this cyto-genomic approach is the improved alignment of the sequence and linkage maps with cytogenetic features such as the centromere, telomeres, p and q arms, and staining patterns indicating GC versus AT content.

The chicken telomerase reverse transcriptase (chTERT): molecular and cytogenetic characterization with a comparative analysis.

Telomerase activity is essential for maintaining the termini of linear chromosomes. Telomerase consists of both a RNA and a specialized reverse transcriptase. Our objective for this study was to determine the molecular and cytogenetic features of the chicken telomerase reverse transcriptase (chTERT) gene and protein. The TERT mRNA from gastrula stage embryos was found to be 4497 bp in length, translating into a protein of 1346 amino acids (aa). The chTERT protein shares 45% aa identity with human TERT (hTERT). A distinctive feature of chTERT, as compared to human and other vertebrate TERTs, is the larger size of the protein due mainly to a considerably longer N-terminal flexible linker region (144 aa longer than in human). Chicken TERT was mapped to chromosome 2q21 near an interstitial telomere site. Several transcription factor binding motifs in the 5' flanking/promoter region of chTERT were similar to those found associated with hTERT (E-box, Ik1, MAZ, Sp1 sites), whereas several c-Myb sites were found associated with chTERT only and c-Ets-2 and WT1 were associated with hTERT only. Results presented here should promote structure-function studies of chTERT, as well as contribute to the comparative analysis of TERT regulation and function in vertebrates utilizing the telomere clock mechanism to different degrees.

Molecular and cytogenetic organization of the 5S ribosomal DNA array in chicken (Gallus gallus).

The 5S ribosomal (r) RNA genes encode a small (approximately 120-bp) highly-conserved component of the large ribosomal subunit. The objective of the present research was to study the molecular and cytogenetic organization of the chicken 5S rDNA. A predominant 2.2-kb gene (5Salpha) consisting of a coding and intergenic spacer (IGS) region was identified in ten research and commercial populations. A variant gene repeat of 0.6kb (5Sbeta) was observed in some of the populations. Genetic linkage analysis and cytogenetic localization by fluorescence in-situ hybridization assigned the 5S rDNA to chromosome 9. The 5S rDNA array was determined to be 80.2 +/- 7.0 kb upon electrophoretic sizing following EcoRV digestion. Sequence analysis of 5Salpha IGS regions revealed considerable conservation between chicken subspecies (98.4% identity) as well as homology with vertebrate Pol III promoter and regulatory sequence motifs. Minor intraindividual sequence variation within 1000 bp of IGS was observed in four cloned Red Jungle Fowl (Gallus gallus gallus) 5Salpha repeats (95.5% identity in this region). Sequence comparisons between IGS regions of 5Salpha and 5Sbeta genes indicated two short continuous (>20bp) and many short non-continuous homologous regions as well as other conserved features such as promoter and termination motifs.

Removal of PCR inhibitors from soil DNA by chemical flocculation.

Extracting high-purity DNA directly from soil has become essential for the study of microorganisms in environmental samples. However, many soils contain compounds that inhibit enzymes involved in manipulating DNA. In this study, chemical flocculation using multivalent cations was investigated as a potential method for eliminating soil-based inhibitors during the extraction process. The addition of AlNH(4)(SO(4))(2) during extraction significantly reduced the co-purification of PCR inhibitors with minimal loss of DNA yield.