Translating Research into Practice: HEADS UP and K-12 Science Inspiring Health and Careers in Youth

Introduction: HEADS UP {Health Education And Discovering Science while Unlocking Potential} aims to improve health literacy and increase student interest in health science careers by providing cutting-edge content from world-renowned researchers in the Texas Medical Center and beyond to the K-12 school community. [See PDF for complete abstract]

Molecular basis of Corynebacterium diphtheriae virulence and infection in the Caenorhabditis elegans model host

Corynebacterium diphtheriae is the causative agent of cutaneous and pharyngeal diphtheria in humans. While lethality is certainly caused by diphtheria toxin, corynebacterial colonization may primarily require proteinaceous fibers called pili, which mediate adherence to specific tissues. The type strain of C. diphtheriae possesses three distinct pilus structures, namely the SpaA, SpaD, and SpaH-type pili, which are encoded by three distinct pilus gene clusters. The pilus is assembled onto the bacterial peptidoglycan by a specific transpeptidase enzyme called sortase. Although the SpaA pili are shown to be specific for pharyngeal cells in vitro, little is known about functions of the three pili in bacterial pathogenesis. This is mainly due to lack of in vivo models of corynebacterial infection. As an alternative to mouse models as mice do not have functional receptors for diphtheria toxin, in this study I use Caenorhabditis elegans as a model host for C. diphtheriae. A simple C. elegans model would be beneficial in determining the specific role of each pilus-type and the literature suggests that C. elegans infection model can be used to study a variety of bacterial species giving insight into bacterial virulence and host-pathogen interactions. My study examines the hypothesis that pili and toxin are major virulent determinants of C. diphtheriae in the C. elegans model host.

Latino Teen Pregnancy in Texas: Prevalence, Prevention, and Policy

Texas is home to over one million Latino teens who are at risk for negative reproductive health outcomes, such as teen pregnancy and STIs. Teen pregnancy disproportionately impacts the health of Latino teens in Texas and places them at risk of continued high rates of poverty, school dropout, and unemployment unless Texas makes a concerted effort to reduce its teen pregnancy rate. The birth rate among Latina girls is astonishing: 98 per 1000 Latinas (aged 15-19) are giving birth. This translates to over 32,000 births each year among Latina teens, costing almost $98 million in direct medical expenditures and well over $638 million if other costs are included. Most teens become sexually experienced while they are of school age, which translates to an estimated 414,583 sexually experienced Latino students attending Texas public schools. Of these Latino youth, 237,466 report being currently sexually active, and 89,000 report having had four or more sexual partners in their lifetime. While causes of teen pregnancy are complex, the solutions to teen pregnancy are known. Texas needs an effective, comprehensive approach to address the sexual health needs of Texas Latino youth that includes: statewide implementation and monitoring of evidence-based sex education for middle school and high school students, access to reproductive health services for students who are already sexually experienced, and widespread training on adolescent sexual health for teachers, service providers, and parents. By tackling teen pregnancy, we can positively impact the future and well-being of not only Latinos, but of all Texans, and subsequently can contribute to the social and economic success of Texas.

Human Trafficking: What is the Role of the Health Care Provider?

The article will address the global and local issue of human trafficking. An estimated 20,000 people are trafficked within the U.S. each year. Trafficked people are forced, defrauded and coerced into labor and sexual service for profit of others. Traffickers use individual vulnerabilities and immigration status, language ability and poor understanding of U.S. laws to identify future victims. One case in 2005 in Texas resulted in 100 victims being identified, none of whom revealed themselves to health care professionals. Health care professionals need contemporary and updated information and resources about health risks, screening methods, and identification of trafficked persons. Readers will learn about common medical problems experienced by individuals who are leaving trafficking situations. Legal and health care intersections will be explored. Implementation of a response protocol to assist those who may currently be enslaved will be introduced. Real case examples from trafficking survivors will be presented and discussed. Participants will learn how to reach out, look beneath the surface, provide assistance, and access resources to help victims, and gain a better understanding of the health challenges faced by trafficked victims.

Morphoproteomic analysis reveals an overexpressed and constitutively activated phospholipase D1-mTORC2 pathway in endometrial carcinoma.

The mammalian target of rapamycin (MTOR) assembles into two distinct complexes: mTOR complex 1 (mTORC1) is predominantly cytoplasmic and highly responsive to rapamycin, whereas mTOR complex 2 (mTORC2) is both cytoplasmic and nuclear, and relatively resistant to rapamycin. mTORC1 and mTORC2 phosphorylatively regulate their respective downstream effectors p70S6K/4EBP1, and Akt. The resulting activated mTOR pathways stimulate protein synthesis, cellular proliferation, and cell survival. Moreover, phospholipase D (PLD) and its product, phosphatidic acid (PA) have been implicated as one of the upstream activators of mTOR signaling. In this study, we investigated the activation status as well as the subcellular distribution of mTOR, and its upstream regulators and downstream effectors in endometrial carcinomas (ECa) and non-neoplastic endometrial control tissue. Our data show that the mTORC2 activity is selectively elevated in endometrial cancers as evidenced by a predominant nuclear localization of the activated form of mTOR (p-mTOR at Ser2448) in malignant epithelium, accompanied by overexpression of nuclear p-Akt (Ser473), as well as overexpression of vascular endothelial growth factor (VEGF)-A isoform, the latter a resultant of target gene activation by mTORC2 signaling via hypoxia-inducible factor (HIF)-2alpha. In addition, expression of PLD1, one of the two major isoforms of PLD in human, is increased in tumor epithelium. In summary, we demonstrate that the PLD1/PA-mTORC2 signal pathway is overactivated in endometrial carcinomas. This suggests that the rapamycin-insensitive mTORC2 pathway plays a major role in endometrial tumorigenesis and that therapies designed to target the phospholipase D pathway and components of the mTORC2 pathway should be efficacious against ECa.

Role of acetyl-phosphate in activation of the Rrp2-RpoN-RpoS pathway in Borrelia burgdorferi.

Borrelia burgdorferi, the Lyme disease spirochete, dramatically alters its transcriptome and proteome as it cycles between the arthropod vector and mammalian host. During this enzootic cycle, a novel regulatory network, the Rrp2-RpoN-RpoS pathway (also known as the σ(54)-σ(S) sigma factor cascade), plays a central role in modulating the differential expression of more than 10% of all B. burgdorferi genes, including the major virulence genes ospA and ospC. However, the mechanism(s) by which the upstream activator and response regulator Rrp2 is activated remains unclear. Here, we show that none of the histidine kinases present in the B. burgdorferi genome are required for the activation of Rrp2. Instead, we present biochemical and genetic evidence that supports the hypothesis that activation of the Rrp2-RpoN-RpoS pathway occurs via the small, high-energy, phosphoryl-donor acetyl phosphate (acetyl∼P), the intermediate of the Ack-Pta (acetate kinase-phosphate acetyltransferase) pathway that converts acetate to acetyl-CoA. Supplementation of the growth medium with acetate induced activation of the Rrp2-RpoN-RpoS pathway in a dose-dependent manner. Conversely, the overexpression of Pta virtually abolished acetate-induced activation of this pathway, suggesting that acetate works through acetyl∼P. Overexpression of Pta also greatly inhibited temperature and cell density-induced activation of RpoS and OspC, suggesting that these environmental cues affect the Rrp2-RpoN-RpoS pathway by influencing acetyl∼P. Finally, overexpression of Pta partially reduced infectivity of B. burgdorferi in mice. Taken together, these findings suggest that acetyl∼P is one of the key activating molecule for the activation of the Rrp2-RpoN-RpoS pathway and support the emerging concept that acetyl∼P can serve as a global signal in bacterial pathogenesis.

Application of in vivo induced antigen technology (IVIAT) to Bacillus anthracis.

In vivo induced antigen technology (IVIAT) is an immuno-screening technique that identifies bacterial antigens expressed during infection and not during standard in vitro culturing conditions. We applied IVIAT to Bacillus anthracis and identified PagA, seven members of a N-acetylmuramoyl-L-alanine amidase autolysin family, three P60 family lipoproteins, two transporters, spore cortex lytic protein SleB, a penicillin binding protein, a putative prophage holin, respiratory nitrate reductase NarG, and three proteins of unknown function. Using quantitative real-time PCR comparing RNA isolated from in vitro cultured B. anthracis to RNA isolated from BALB/c mice infected with virulent Ames strain B. anthracis, we confirmed induced expression in vivo for a subset of B. anthracis genes identified by IVIAT, including L-alanine amidases BA3767, BA4073, and amiA (pXO2-42); the bacteriophage holin gene BA4074; and pagA (pXO1-110). The exogenous addition of two purified putative autolysins identified by IVIAT, N-acetylmuramoyl-L-alanine amidases BA0485 and BA2446, to vegetative B. anthracis cell suspensions induced a species-specific change in bacterial morphology and reduction in viable bacterial cells. Many of the proteins identified in our screen are predicted to affect peptidoglycan re-modeling, and our results support significant cell wall structural remodeling activity during B. anthracis infection. Identification of L-alanine amidases with B. anthracis specificity may suggest new potential therapeutic targets.

Dispelling the Myth: What Parents Really Think about Sex Education in Schools

Background: School-based sex education is effective in reducing risky sexual behavior among adolescents that may lead to unintended pregnancies and sexually transmitted infections. However, most sex education policies in the US do not support evidence-based programs. Understanding parental attitudes around sex education is crucial to overcoming perceived barriers to implementing school-based sex education. Little research has been published on the opinions of parents in Texas, which accounts for 12% of the nation’s teen births. Purpose: The purpose of this study was to examine whether Texas parents favor teaching sex education in schools, in what grades they think sex education should be taught, what content they think should be taught, and who they think should make decisions regarding sex education. Methods: We commissioned a telephone survey of parents of children 18 years or younger in Harris County, Texas. Survey questions assessed demographic characteristics and opinions about sex education. We used chi-square tests to examine differences across sociodemographic characteristics. Results: 1,201 parents completed the survey. The majority of parents (80%) responded that sex education should begin in middle school or earlier, and two-thirds said that it should include information about condoms and contraception. Hispanic parents showed the highest support for teaching sex education and providing medically accurate information on condoms and contraception in middle school or earlier. Conclusion: Parents in Harris County overwhelmingly support sex education that includes medically accurate information about condoms and contraception beginning before high school. These data provide evidence to change sex education policies to better reflect parental opinions.

Choosing and Maintaining Programs for Sex Education in Schools: The CHAMPSS Model

Background: Despite effective solutions to reduce teen birth rates, Texas teen birth rates are among the highest in the nation. School districts can impact youth sexual behavior through implementation of evidence-based programs (EBPs); however, teen pregnancy prevention is a complex and controversial issue for school districts. Subsequently, very few districts in Texas implement EBPs for pregnancy prevention. Additionally, school districts receive little guidance on the process for finding, adopting, and implementing EBPs. Purpose: The purpose of this report is to present the CHoosing And Maintaining Programs for Sex education in Schools (CHAMPSS) Model, a practical and realistic framework to help districts find, adopt, and implement EBPs. Methods: Model development occurred in four phases using the core processes of Intervention Mapping: 1) knowledge acquisition, 2) knowledge engineering, 3) model representation, and 4) knowledge development. Results: The CHAMPSS Model provides seven steps, tailored for school-based settings, which encompass phases of assessment, preparation, implementation, and maintenance: Prioritize, Asses, Select, Approve, Prepare, Implement, and Maintain. Advocacy and eliciting support for adolescent sexual health are also core elements of the model. Conclusion: This systematic framework may help schools increase adoption, implementation, and maintenance for EBPs.

Sexual Health Education from the Perspective of School Staff: Implications for Adoption and Implementation of Effective Programs in Middle School

Introduction: US teens are having sex early; however, the vast majority of schools do not implement evidence-based sexual health education (SHE) programs that could delay sexual behavior and/or reduce risky behavior. This study examines middle school staff’s knowledge, attitudes, barriers, self-efficacy, and perceived support (psychosocial factors known to influence SHE program adoption and implementation). Methods: Professional school staff from 33 southeast Texas middle schools completed an internet or paper-based survey. Prevalence estimates for psychosocial variables were computed for the total sample. Chi-square and t-test analyses examined variation by demographic factors. Results: Almost 70% of participants were female, 37% white, 42% black, 16% Hispanic; 20% administrators, 15% nurses/counselors, 31% non-physical education/non-health teachers, 28% physical education/health teachers; mean age = 42.78 years (SD = 10.9). Over 90% favored middle school SHE, and over 75% reported awareness of available SHE curricula or policies. More than 60% expressed confidence for discussing SHE. Staff perceived varying levels of administrator (28%-56%) support for SHE and varying levels of support for comprehensive sex education from outside stakeholders (e.g., parents, community leaders) (42%-85%). Overall, results were more favorable for physical education/health teachers, nurses/counselors, and administrators (when compared to non-physical education/non-health teachers) and individuals with experience teaching SHE. Few significant differences were observed by other demographic factors. Conclusions: Overall, study results were extremely positive, which may reflect a high level of readiness among school staff for adopting and implementing effective middle school SHE programs. Study results highlight the importance of several key action items for schools.

A Tale of Two States: What We Learn from California and Texas

Teen birth rates and teen pregnancy prevention strategies vary widely across individual states in the US, which has the highest overall teen birth rate among developed nations. California and Texas, the two most populous states currently accounting for a quarter of all teen births, have taken very different approaches to addressing adolescent reproductive health. This case study examines the racial/ethnic composition and socioeconomic factors of these two states from 1981 to 2008. State programs and policies implemented between 1991 and 2008 as well as changes in access to contraception and public–private partnerships are discussed. Based on the lessons learned from California, a similar multifaceted campaign in Texas may be effective in reducing teen births.

Adolescent Sexual Behavior: Examining Data from Texas and the US

Background: The US has higher rates of teen births and sexually transmitted infections (STI) than other developed countries. Texas youth are disproportionately impacted. Purpose: To review local, state, and national data on teens’ engagement in sexual risk behaviors to inform policy and practice related to teen sexual health. Methods: 2009 middle school and high school Youth Risk Behavior Survey (YRBS) data, and data from All About Youth, a middle school study conducted in a large urban school district in Texas, were analyzed to assess the prevalence of sexual initiation, including the initiation of non-coital sex, and the prevalence of sexual risk behaviors among Texas and US youth. Results: A substantial proportion of middle and high school students are having sex. Sexual initiation begins as early as 6th grade and increases steadily through 12th grade with almost two-thirds of high school seniors being sexually experienced. Many teens are not protecting themselves from unintended pregnancy or STIs – nationally, 80% and 39% of high school students did not use birth control pills or a condom respectively the last time they had sex. Many middle and high school students are engaging in oral and anal sex, two behaviors which increase the risk of contracting an STI and HIV. In Texas, an estimated 689,512 out of 1,327,815 public high school students are sexually experienced – over half (52%) of the total high school population. Texas students surpass their US peers in several sexual risk behaviors including number of lifetime sexual partners, being currently sexually active, and not using effective methods of birth control or dual protection when having sex. They are also less likely to receive HIV/AIDS education in school. Conclusion: Changes in policy and practice, including implementation of evidence-based sex education programs in middle and high schools and increased access to integrated, teen-friendly sexual and reproductive health services, are urgently needed at the state and national levels to address these issues effectively.

Primary lung cancer after breast cancer: The role of radiation therapy and cigarette smoking

The magnitude of the interaction between cigarette smoking, radiation therapy, and primary lung cancer after breast cancer remains unresolved. This case control study further examines the main and joint effects of cigarette smoking and radiation therapy (XRT) among breast cancer patients who subsequently developed primary lung cancer, at The University of Texas M. D. Anderson Cancer Center (MDACC) in Houston, Texas. Cases (n = 280) were women diagnosed with primary lung cancer between 1955 and 1970, between 30–89 years of age, who had a prior history of breast cancer, and were U.S. residents. Controls (n = 300) were randomly selected from 37,000 breast cancer patients at MDACC and frequency matched to cases on age at diagnosis (in 5-year strata), ethnicity, year of breast cancer diagnosis (in 5-year strata), and had survived at least as long as the time interval for lung cancer diagnosis in the cases. Stratified analysis and unconditional logistic regression modeling were used to calculate the main and joint effects of cigarette smoking and radiation treatment on lung cancer risk. Medical record review yielded smoking information on 93% of cases and 84% of controls, and among cases 45% received XRT versus 44% of controls. Smoking increased the odds of lung cancer in women who did not receive XRT (OR = 6.0, 95%CI, 3.5–10.1) whereas XRT was not associated with increased odds (OR = 0.5, 95%CI, 0.2–1.1) in women who did not smoke. Overall the odds ratio for both XRT and smoking together compared with neither exposure was 9.00 (9 5% CI, 5.1–15.9). Similarly, when stratifying on laterality of the lung cancer in relation to the breast cancer, and when the time interval between breast and lung cancers was >10 years, there was an increased odds for both smoking and XRT together for lung cancers on the same side as the breast cancer (ipsilateral) (OR = 11.5, 95% CI, 4.9–27.8) and lung cancers on the opposite side of the breast cancer (contralateral) (OR= 9.6, 95% CI, 2.9–0.9). After 20 years the odds for the ipsilateral lung were even more pronounced (OR = 19.2, 95% CI, 4.2–88.4) compared to the contralateral lung (OR = 2.6, 95% CI, 0.2–2.1). In conclusion, smoking was a significant independent risk factor for lung cancer after breast cancer. Moreover, a greater than multiplicative effect was observed with smoking and XRT combined being especially evident after 10 years for both the ipsilateral and contralateral lung and after 20 years for the ipsilateral lung. ^

Capsule gene regulation in Bacillus anthracis and implications for virulence

The poly-D-glutamic acid capsule of Bacillus anthracis is considered essential for lethal anthrax disease. Yet investigations of capsule function have been limited primarily to attenuated B. anthracis strains lacking certain genetic elements. In work presented in this thesis, I constructed and characterized a genetically complete (pXO1 + pXO2+) B. anthracis strain (UT500) and isogenic mutants deleted for two previously identified capsule gene regulators, atxA and acpA, and a newly-identified regulator, acpB. Results of transcriptional analysis and microscopy revealed that atxA controls expression of the first gene of the capsule biosynthesis operon, capB, via positive transcriptional regulation of acpA and acpB. acpA and acpB appear to be partial functional homologs. Deletion of either gene alone has little effect on capsule synthesis. However, a mutant deleted for both acpA and acpB is noncapsulated. Thus, in contrast to previously published models, my results suggest that atxA is the master regulator of cap gene expression in a genetically complete strain. A detailed transcriptional analysis of capB and the regulatory genes was performed to establish the effects of the regulators and CO2/bicarbonate on specific mRNAs of target genes. CO2/bicarbonate is a well-established signal for B. anthracis capsule synthesis in culture. Taqman RT-PCR results indicated that growth in the presence of elevated CO2 greatly increased expression of acpA, acpB and capB but not atxA. 5′ end mapping of capB and acpA revealed atxA-regulated and atxA-independent transcriptional start sites for both genes. All atxA-regulated start sites were also CO2-regulated. A single atxA-independent start site was identified 5 ′ of acpB. However, RT-PCR analysis indicated that capD and acpB are co-transcribed. Thus, it is likely that atxA-mediated control of acpB expression occurs via transcriptional activation of the atxA-regulated start sites of capB. Finally, I examined the contribution of the B. anthracis capsule to virulence. The virulence of the parent strain, mutants deleted for the capsule biosynthesis genes ( capBCAD), and mutants missing the capsule regulator genes was compared using a mouse model for inhalation anthrax. The data indicate that in this model, capsule is essential for virulence. Mice survived infection with the noncapsulated capBCAD and acpA acpB mutants. These mutants initiated germination in the lung, but did not disseminate to the spleen. The acpA mutant had an LD50 value similar to the parent strain and was able to disseminate and cause lethal infection. Unexpectedly, the acpB mutant had a higher LD 50 and a reduced ability to disseminate. During in vitro culture, the acpB single mutant produces capsule and toxin similar to the parent strain. It is likely that acpB regulates the expression of downstream genes that contribute to the virulence of B. anthracis. ^

B cells in ADA deficiency

Deficiency of the enzyme adenosine deaminase (ADA) results in severe lymphopenia in humans. Mice with an inactivating mutation in the ADA gene also exhibit profound lymphopenia, as well as pulmonary insufficiency and ribcage abnormalities. In fact, the mouse model has a phenotype that is remarkably similar to that of the human disease, making the mice valuable tools for unraveling the mechanism of lymphocyte destruction in absence of this housekeeping gene. T cell deficiency in ADA deficiency has been extensively studied by others, revealing a block in early thymocyte development. In contrast, our studies revealed that early B cell development in the bone marrow is normal. ADA-deficient mice, however, exhibit profound defects in germinal center formation, preventing antigen-dependent B cell maturation in the spleen. ADA-deficient spleen B cells display significant defects in proliferation and activation signaling, and produce more IgM than their normal counterparts, suggesting that extrafollicular plasmablasts are overrepresented. B cells from ADA-deficient mouse spleens undergo apoptosis more readily than those from normal mouse spleens. Levels of ADA's substrates, adenosine and 2′-deoxyadenosine, are elevated in both bone marrow and spleen in ADA-deficient mice. S ′-adenosyihomoeysteine hydrolase (SAH hydrolase) activity is significantly inhibited in both locales, as well. dATP levels, though, are only elevated in spleen, where B cell development is impaired, and not in bone marrow, where B cell ontogeny is normal. This finding points to dATP as the causative agent of lymphocyte death in ADA deficiency. ADA deficiency results in inhibition of the enzyme ribonucleotide reductase, thereby depleting nucleoside pools needed for DNA repair. Another mouse model that lacks a functional gene encoding a protein involved in DNA repair and/or cell cycle checkpoint regulation, p53-binding protein 1, exhibits blocks in T and B cell development that are similar to those seen in ADA-deficient mice. Unraveling the mechanisms of lymphocyte destruction in ADA deficiency may further understanding of lymphocyte biology, facilitate better chemotherapeutic treatment for lymphoproliferative diseases, and improve gene and enzyme therapy regimens attempted for ADA deficiency. ^