Managing hundreds of improvement teams.

Recognizing the notable scale of USAID Applying Science to Strengthen and Improve Systems (ASSIST) Project activities and sizable number of improvement teams, which in some cases is close to 1,000 improvement teams managed in one country at a point in time, we sought to answer the questions: How do we manage hundreds of improvement teams in one country alone? How do we manage more than 4,000 improvement teams globally? The leaders of our improvement programs manage such efforts as though they are second-nature, without pointing to the specific skills and strategies needed to manage thousands of teams. This paper was developed to capture the lessons, considerations, and insights shared in discussions with leaders on the USAID ASSIST Project, including country Chiefs of Party and Regional Directors. More specifically, this paper seeks to describe what is involved in scaling up and managing large numbers of improvement teams. Through focus group discussions and individual interviews, participants discussed the key skills, strategies, and lessons needed to successfully manage large numbers of teams on the USAID ASSIST Project. We concluded that the six key components in managing large numbers of teams are 1) leadership; 2) management structures and capacities; 3) clear and open communication; 4) shared learning, collaboration, and support; 5) ownership, engagement, and empowerment; and 6) partnerships. We further analyzed these six components as being interrelated to one another based on the relationship between culture, strategy, and technique in implementing quality improvement activities.

Evidence for Hox-specified positional identities in adult vasculature.

BACKGROUND: The concept of specifying positional information in the adult cardiovascular system is largely unexplored. While the Hox transcriptional regulators have to be viewed as excellent candidates for assuming such a role, little is known about their presumptive cardiovascular control functions and in vivo expression patterns. RESULTS: We demonstrate that conventional reporter gene analysis in transgenic mice is a useful approach for defining highly complex Hox expression patterns in the adult vascular network as exemplified by our lacZ reporter gene models for Hoxa3 and Hoxc11. These mice revealed expression in subsets of vascular smooth muscle cells (VSMCs) and endothelial cells (ECs) located in distinct regions of the vasculature that roughly correspond to the embryonic expression domains of the two genes. These reporter gene patterns were validated as authentic indicators of endogenous gene expression by immunolabeling and PCR analysis. Furthermore, we show that persistent reporter gene expression in cultured cells derived from vessel explants facilitates in vitro characterization of phenotypic properties as exemplified by the differential response of Hoxc11-lacZ-positive versus-negative cells in migration assays and to serum. CONCLUSION: The data support a conceptual model of Hox-specified positional identities in adult blood vessels, which is of likely relevance for understanding the mechanisms underlying regional physiological diversities in the cardiovascular system. The data also demonstrate that conventional Hox reporter gene mice are useful tools for visualizing complex Hox expression patterns in the vascular network that might be unattainable otherwise. Finally, these mice are a resource for the isolation and phenotypic characterization of specific subpopulations of vascular cells marked by distinct Hox expression profiles.

Vignettes: Diverse library staff offering diverse bioinformatics services.

OBJECTIVES: The paper gives examples of the bioinformatics services provided in a variety of different libraries by librarians with a broad range of educational background and training. METHODS: Two investigators sent an email inquiry to attendees of the "National Center for Biotechnology Information's (NCBI) Introduction to Molecular Biology Information Resources" or "NCBI Advanced Workshop for Bioinformatics Information Specialists (NAWBIS)" courses. The thirty-five-item questionnaire addressed areas such as educational background, library setting, types and numbers of users served, and bioinformatics training and support services provided. Answers were compiled into program vignettes. DISCUSSION: The bioinformatics support services addressed in the paper are based in libraries with academic and clinical settings. Services have been established through different means: in collaboration with biology faculty as part of formal courses, through teaching workshops in the library, through one-on-one consultations, and by other methods. Librarians with backgrounds from art history to doctoral degrees in genetics have worked to establish these programs. CONCLUSION: Successful bioinformatics support programs can be established in libraries in a variety of different settings and by staff with a variety of different backgrounds and approaches.

Evidence that the satin hair mutant gene Foxq1 is among multiple and functionally diverse regulatory targets for Hoxc13 during hair follicle differentiation.

It is increasingly evident that the molecular mechanisms underlying hair follicle differentiation and cycling recapitulate principles of embryonic patterning and organ regeneration. Here we used Hoxc13-overexpressing transgenic mice (also known as GC13 mice), known to develop severe hair growth defects and alopecia, as a tool for defining pathways of hair follicle differentiation. Gene array analysis performed with RNA from postnatal skin revealed differential expression of distinct subsets of genes specific for cells of the three major hair shaft compartments (cuticle, cortex, and medulla) and their precursors. This finding correlates well with the structural defects observed in each of these compartments and implicates Hoxc13 in diverse pathways of hair follicle differentiation. The group of medulla-specific genes was particularly intriguing because this included the developmentally regulated transcription factor-encoding gene Foxq1 that is altered in the medulladefective satin mouse hair mutant. We provide evidence that Foxq1 is a downstream target for Hoxc13 based on DNA binding studies as well as co-transfection and chromatin immunoprecipitation assays. Expression of additional medulla-specific genes down-regulated upon overexpression of Hoxc13 requires functional Foxq1 as their expression is ablated in hair follicles of satin mice. Combined, these results demonstrate that Hoxc13 and Foxq1 control medulla differentiation through a common regulatory pathway. The apparent regulatory interactions between members of the mammalian Hox and Fox gene families shown here may establish a paradigm for "cross-talk" between these two conserved regulatory gene families in different developmental contexts including embryonic patterning as well as organ development and renewal.

Epididymal cysteine-rich secretory protein 1 encoding gene is expressed in murine hair follicles and downregulated in mice overexpressing Hoxc13.

Members of the Hox gene family of transcriptional regulators are believed to play essential roles in hair follicle differentiation, although little is known about the molecular mechanisms mediating these putative control functions. Transgenic mice overexpressing Hoxc13 in hair follicles (GC13 mice) exhibit complex phenotypic alterations including hair shaft defects and alopecia, as well as severe epidermal abnormalities. To identify some of the genetic pathways affected by Hoxc13 overexpression in hair, we performed large-scale differential gene expression analysis on the skin of 5-d GC13 versus normal FVB mice using DNA chip assays. A surprising result of these experiments was the identification of the epididymal cysteine-rich secretory protein 1 (Crisp1) gene as one of the genes with the greatest expression differential, in this case with greater than 20-fold downregulation in skin from GC13 mice. Crisp1 encodes a secreted protein that has originally been found to be abundantly expressed in the epididymis, where it plays a role in sperm maturation. We have localized Crisp1 mRNA in 5-d postnatal murine scapular skin by in situ hybridization, showing its expression to be restricted to the medulla of the hair shaft. Furthermore, we provide evidence for specific interaction of Hoxc13 with at least one cognate binding site found in the Crisp1 promoter region, thus supporting the concept of a Hoxc13/Crisp1 regulatory relationship. In summary, these data establish the hair as a novel site for Crisp1 expression where its functional role remains to be determined.

Krtap16, characterization of a new hair keratin-associated protein (KAP) gene complex on mouse chromosome 16 and evidence for regulation by Hoxc13.

Intermediate filament (IF) keratins and keratin-associated proteins (KAPs) are principal structural components of hair and encoded by members of multiple gene families. The severe hair growth defects observed upon aberrant expression of certain keratin and KAP genes in both mouse and man suggest that proper hair growth requires their spatio-temporally coordinated activation. An essential prerequisite for studying these cis-regulatory mechanisms is to define corresponding gene families, their genomic organization, and expression patterns. This work characterizes eight recently identified high glycine/tyrosine (HGT)-type KAP genes collectively designated Krtap16-n. These genes are shown to be integrated into a larger KAP gene domain on mouse chromosome 16 (MMU16) that is orthologous to a recently described HGT- and high sulfur (HS)-type KAP gene complex on human chromosome 21q22.11. All Krtap16 genes exhibit strong expression in a narrowly defined pattern restricted to the lower and middle cortical region of the hair shaft in both developing and cycling hair. During hair follicle regression (catagen), expression levels decrease until expression is no longer detectable in follicles at resting stage (telogen). Since isolation of the Krtap16 genes was based on their differential expression in transgenic mice overexpressing the Hoxc13 transcriptional regulator in hair, we examined whether bona fide Hoxc13 binding sites associated with these genes might be functionally relevant by performing electrophoretic mobility shift assays (EMSAs). The data provide evidence for sequence-specific interaction between Hoxc13 and Krtap16 genes, thus supporting the concept of a regulatory relationship between Hoxc13 and these KAP genes.

A testing algorithm for determination of HER2 status in patients with breast cancer.

The HER2 gene is amplified and overexpressed in 25-30% of breast carcinomas. Assessment of HER2 status for prognosis and treatment of breast cancer patients can be performed by immunohistochemistry and/or fluorescence in situ hybridization (FISH). To develop a testing algorithm for HER2 in breast cancers, we used FISH analysis to determine the HER2 gene copy number and immunostaining to detect the p185 protein. Interlaboratory, interobserver, and intermethod variabilities of immunohistochemistry were assessed. In 24 invasive breast carcinomas, the indices of HER2 status obtained by FISH and by a reference laboratory's DAKO HercepTest (immunostain) gave an overall concordance of 96%. The reference laboratory's stained slides were re-interpreted by an in-house panel of pathologists; the interpretation differed in one case. The panel's interpretations were concordant with the FISH results in all 24 cases. Interobserver variability for the panel's immunohistochemistry interpretations was assessed using three different immunostaining methods on 70 slides. The numerical (0-1+, 2+, 3+) scores showed greater variability among observers than did the overall positive/negative results. One pathologist reported inconsistent results in >30% of the slides evaluated. Borderline scoring of 1-2+ was reported in 18 slides (23%) by at least one observer. Incongruent interobserver immunohistochemistry scores, leading to different positive and negative interpretations, were obtained with 5 slides (7%). The majority of consensus positive cases exhibited strong membrane staining (3+). The majority of consensus negative cases scored as 0. Based on these observations, we developed a testing algorithm that maximizes the benefits of FISH and immunohistochemistry, providing physicians with accurate results for appropriate clinical care.