Identifying Bacterial and Host Factors Involved in the Interaction of Mycobacterium bovis with the Bovine Innate Immune Cells.

Bovine tuberculosis is an important animal and zoonotic disease caused by Mycobacterium bovis. The innate immune response is the first line of defense against pathogens and is also crucial for the development of an efficient adaptive immune response. In this study we used an in vitro co-culture model of antigen presenting cells (APC) and autologous lymphocytes derived from peripheral blood mononuclear cells to identify the cell populations and immune mediators that participate in the development of an efficient innate response capable of controlling the intracellular replication of M. bovis. After M. bovis infection, bovine immune cell cultures displayed upregulated levels of iNOS, IL-22 and IFN-γ and the induction of the innate immune response was dependent on the presence of differentiated APC. Among the analyzed M. bovis isolates, only a live virulent M. bovis isolate induced an efficient innate immune response, which was increased upon stimulation of cell co-cultures with the M. bovis culture supernatant. Moreover, we demonstrated that an allelic variation of the early secreted protein ESAT-6 (ESAT6 T63A) expressed in the virulent strain is involved in this increased innate immune response. These results highlight the relevance of the compounds secreted by live M. bovis as well as the variability among the assessed M. bovis strains to induce an efficient innate immune response.

Determination of dehydroepiandrosterone and its biologically active oxygenated metabolites in human plasma evinces a hormonal imbalance during HIV-TB coinfection.

An estimated one third of the world's population is affected by latent tuberculosis (TB), which once active represents a leading cause of death among infectious diseases. Human immunodeficiency virus (HIV) infection is a main predisposing factor to TB reactivation. Individuals HIV-TB co-infected develop a chronic state of inflammation associated with hypothalamic-pituitary-adrenal (HPA) axis dysregulation. This results in a hormonal imbalance, disturbing the physiological levels of cortisol and dehydroepiandrosterone (DHEA). DHEA and its oxygenated metabolites androstenediol (AED), androstenetriol (AET) and 7-oxo-DHEA are immunomodulatory compounds that may regulate physiopathology in HIV-TB co-infection. In order to study possible changes in plasma levels of these hormones, we developed an approach based on high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). To our knowledge, this represents the first report of their simultaneous measurement in HIV-TB individuals and the comparison with healthy donors, obtaining statistically higher plasma levels of DHEA, AET and 7-oxo-DHEA in patients. Moreover, we found that concentrations of 7-oxo-DHEA positively correlated with absolute CD4+ T cell counts, nadir CD4+ T cell values and with individuals who presented TB restricted to the lungs. This research contributes to understanding the role of these hormones in HIV-TB and emphasizes the importance of deepening their study in this context.

Validación de un método rápido en orina por lC/MS/MS para la búsqueda de compuestos empleados en sumisión química y su aplicación a muestras reales / Validation of a fast screening method in urine by lC/MS/MS for compounds used in drug-facilitated crime (DFC) and its application to real samples

La Oficina de Naciones Unidas contra la Droga y el Delito (UNODC) en 2011 señala que "El delito facilitado por drogas (DFD) es una expresión general que abarca la violación y otras agresiones sexuales, el robo con violencia o intimidación, la extorsión de dinero y los malos tratos deliberados de ancianos o niños bajo la influencia de sustancias sicotrópicas". En este trabajo se validó un método cualitativo y rápido a partir de muestras de orina por LC/MS/MS para 39 compuestos comprendidos en los listados de sumisión química. El objetivo fue alcanzar un límite de detección un 50 % por debajo de la concentración propuesta como "Límites mínimos de funcionamiento exigidos (MRPL)" por la UNODC, para poder ser aplicado a muestras reales.

The United Nations Office on Drugs and Crime (UNODC) in 2011, states that "The Drug-facilitated crime (DFC) is a general term that includes rape or other sexual assault, robbery, money extortion, as well as the deliberate maltreatment of the elderly or children under the influence of psychotropic substances". In this work we validated a qualitative and fast method from urine samples by LC/MS/MS for 39 compounds included in the Drug-facilitated crime lists. The aim was to reach a detection limit of 50% below the proposed concentration as "minimum required performance limits (MRPL)" by UNODC in order to be applied in real samples.

Attitudes toward HPV vaccination among low-income and minority parents of sons: a qualitative analysis.

OBJECTIVE: To characterize the attitudes of low-income and minority parents/guardians toward vaccinating sons against human papillomavirus (HPV). METHODS: In 2010-2011, we conducted qualitative interviews with 68 black, 24 white, and 28 Latino parents/guardians of sons. We identified attitudes related to HPV vaccination, vaccine mandates for males and females, and adolescent male sexuality using constructs from the Health Belief Model and methods based in grounded theory. RESULTS: Most participants were concerned that their sons could be exposed to HPV through sexual experimentation and believed that the consequences of HPV infection could be severe; thus, 75% would accept HPV vaccine for their sons. Yet the lack of efficacy and safety information specifically pertaining to males posed barriers. More black (73%) and Latino (86%) than white (44%) participants supported school-entry requirements for HPV vaccination. CONCLUSIONS: Low-income and minority parents/guardians were generally receptive toward vaccinating their sons against HPV; racial/ethnic differences emerged regarding school-entry mandates.

Factors affecting human papillomavirus vaccine use among White, Black and Latino parents of sons.

BACKGROUND: Although human papillomavirus (HPV) vaccination has been available for males since 2009, its uptake remains low. In light of new recommendations for universal vaccination of males, understanding parental attitudes toward this vaccine is important. This study aimed to describe HPV-related knowledge and intention to accept HPV vaccination among White, Black and Latino parents of sons and to assess vaccination rates among their sons. METHODS: We interviewed parents (68 Black, 28 Latino and 24 White; mean age, 43.5) of sons (mean age, 14) attending an urban academic medical center and a community health center. Eligible parents self-identified as White, Black or Latino and spoke English, Spanish or Haitian-Creole. We collected demographic information, knowledge related to HPV vaccination, parents' intent to vaccinate sons and HPV vaccination rates. Descriptive statistics and multivariable logistic regression were used to describe data. RESULTS: Most parents were mothers, married, expressed a religious affiliation and had completed high school or college. Parents had limited knowledge about HPV; White parents were more knowledgeable than Black parents. Most parents (75%) intended to accept HPV vaccination if recommended by physicians; no racial differences were noted. However, only 30% of sons were vaccinated. Logistic regression indicated that internet use was negatively associated with intention to vaccinate. Intention to vaccinate, clinical site of care and having an older son were associated with vaccine receipt. CONCLUSIONS: Although parents in our study had limited understanding of HPV disease in males, most would vaccinate their sons if recommended by their physicians.

A quantitative glycogen assay to verify use of self-administered vaginal swabs.

BACKGROUND: Self-administered swabs are used to sample vaginal contents for a variety of clinical purposes including detection of sexually transmitted infections, condom breakage, and vaginal product use. The goal of this study was to determine whether a quantitative glycogen assay can be used to assess whether a swab has been exposed to the vagina to assure study compliance. STUDY DESIGN: Buccal, skin, or vaginal samples were tested to determine whether a commercial quantitative glycogen assay can differentiate vaginal specimens. In addition, archived remnant de-identified vaginal swabs from clinical trials were tested. Periodic acid-Schiff stain was used to identify glycogen-positive cells as a confirmation test. RESULTS: Glycogen concentrations in eluates of vaginal swabs from reproductive-aged women were significantly higher than those from unused swabs (mean ± SE, 964 ± 135 µg/mL vs. 14.7 ± 2.5 µg/mL, P < 0.001) and swabs exposed to buccal and finger/hand epithelia (40.3 ± 4.8 and 18.5 ± 5.4 µg/mL, P < 0.001). Glycogen concentrations were lower and more variable in vaginal swabs from older perimenopausal/menopausal women (mean ± SE, 235 ± 123, P < 0.01). Semen and sample storage longer than 1 year did not affect glycogen detection. Using a cutoff of 100 µg/mL of glycogen, 30 of 30 vaginal swabs from reproductive-aged women versus 0 of 28 control swabs were positive, for an assay sensitivity of 1 (95% confidence interval, 0.86-1) and specificity of 1 (95% confidence interval, 0.85-1). Periodic acid-Schiff stain correlated with soluble glycogen results but was less specific. CONCLUSIONS: The quantitative glycogen assay provides a simple and inexpensive method to validate the use of self-administered swabs for sampling vaginal contents in clinical studies.

Parents' opinions of mandatory human papillomavirus vaccination: does ethnicity matter?

OBJECTIVE: To explore parents' opinions of school-entry requirements for human papillomavirus (HPV) vaccination. METHODS: We interviewed parents of vaccine-eligible girls attending medical appointments in an urban academic medical center and an affiliated community health center. We used qualitative methods to explore parents' opinions about mandating routine childhood vaccines and HPV vaccine, as well as their feelings about vaccinating their own daughters against HPV. RESULTS: We included 19 Caucasian, 18 African-American, 12 Afro-Caribbean, 3 African, and 21 Latino parents. Nearly all parents had allowed their children to receive routine vaccinations and expressed support for mandating these vaccines. Most parents also vaccinated their daughters against HPV: 100% of Caucasian parents, 90% of African-American parents, 73% of Afro-Caribbean/African parents, and 90% of Latino parents. Only 11% of Caucasian parents supported HPV vaccine mandates, however, compared with 78% of African-American, 60% of Afro-Caribbean/African, and 90% of Latino parents. Immigrants supported mandates more frequently than U.S.-born parents. Most Caucasian parents opposed mandatory HPV vaccination because they believed the HPV vaccine should be an individual decision because the virus can only be spread by sexual contact. African-American, Afro-Caribbean, African, and Latino parents generally viewed mandates as the most effective way to protect their daughters from cervical cancer. Latino parents gave special importance to protecting their daughters from sexually transmitted infections. CONCLUSION: Parents from different racial and ethnic backgrounds expressed unique perspectives about mandatory HPV vaccination. Caucasians were less likely than parents of other races/ethnicities to support vaccine mandates.

Why do low-income minority parents choose human papillomavirus vaccination for their daughters?

OBJECTIVE: To explore low-income minority parents' attitudes, intentions, and actions with regard to human papillomavirus (HPV) vaccination for their daughters. STUDY DESIGN: Semistructured interviews were conducted in English and Spanish with parents of girls aged 11-18 who were attending clinic visits in an urban medical center and a community health center. We assessed intention with formal scales, probed parents' attitudes regarding vaccination with open-ended questions, and reviewed medical records to determine vaccination rates. Data were analyzed using descriptive statistics and qualitative methods. RESULTS: Seventy-six parents participated (43% African American, 28% Latino, and 26% Caucasian). Most were mothers, had completed high school, and described themselves as religious; nearly one-half were immigrants. Intention correlated highly with receipt of the vaccine; 91% of parents intended to vaccinate their daughters against HPV, and 89% of the girls received vaccination within 12 months of the interview. Qualitative analysis revealed that most parents focused on the vaccine's potential to prevent cervical cancer. Some parents expressed concerns about unknown side effects and promotion of unsafe sexual practices, but these concerns did not hinder acceptance in most cases. CONCLUSIONS: The majority of the low-income minority parents surveyed viewed HPV vaccination as a way to protect their daughters from cancer, and thus chose to vaccinate their daughters.