Sulforaphane Suppresses the Growth of Triple-negative Breast Cancer Stem-like Cells In vitro and In vivo.

Triple-negative breast cancer (TNBC) represents the poorest prognosis among all of breast cancer subtypes with no currently available effective therapy. In this study, we hypothesized that sulforaphane, a dietary component abundant in broccoli and its sprouts, can inhibit malignant cell proliferation and tumor sphere formation of cancer stem-like cells (CSC) in TNBC. CSC population was isolated using FACS analysis with the combined stem cell surface markers, CD44+/CD24-/CD49f+ The effect of sulforaphane on a stem-related embryonic oncogene CRIPTO-1/TDGF1 (CR1) was evaluated via ELISA. In vivo, BalbC/nude mice were supplemented with sulforaphane before and after TNBC cell inoculation (daily intraperitoneal injection of 50 mg sulforaphane/kg for 5 and 3 weeks, respectively), and the effects of sulforaphane during mammary tumor initiation and growth were accessed with NanoString gene analysis. We found that sulforaphane can inhibit cell proliferation and mammosphere formation of CSCs in TNBC. Further analysis of gene expression in these TNBC tumor cells revealed that sulforaphane significantly decreases the expression of cancer-specific CR1, CRIPTO-3/TDGF1P3 (CR3, a homologue of CR1), and various stem cell markers including Nanog, aldehyde dehydrogenase 1A1 (ALDH1A1), Wnt3, and Notch4. Our results suggest that sulforaphane may control the malignant proliferation of CSCs in TNBC via Cripto-mediated pathway by either suppressing its expression and/or by inhibiting Cripto/Alk4 protein complex formation. Thus, the use of sulforaphane for chemoprevention of TNBC is plausible and warrants further clinical evaluation.

Valores hematológicos e bioquímicos de pinguins-de-Magalhães em reabilitação no Espírito Santo, sudeste do Brasil / Hematological and biochemical values of Magellanic penguins in rehabilitation in Espírito Santo, southeastern Brazil

Este trabalho apresenta valores hematológicos e bioquímicos de pinguins-de-Magalhães (Spheniscus magellanicus) juvenis que arribaram no Espírito Santo e Rio de Janeiro, litoral sudeste do Brasil, e foram encaminhados a reabilitação. Os valores médios obtidos foram: eritrócitos 2.55±0.54 milhões/µL; hemoglobina 13,37±3,91g/dL; hematócrito 41,5±0,04%; proteína plasmática 6,34±0,81g/dL; leucócitos 16.301±6.402/µL; trombócitos 20.516±4.591 células/µL; volume corpuscular médio (VCM) 172,0±53,0 fL; concentração de hemoglobina corpuscular média (CHCM) 26,41±0,02%; alanina transaminase (ALT) 50,0±17,89 U/L; ácido úrico 8,93±3,0mg/dL; albumina 1,68±0,53g/dL; cálcio 9,7±0,57mg/dL; fósforo 10,39±8,5mg/dL e glicose 211,6±30,3mg/dL. Valores de eritrócitos, VCM e CHCM foram discrepantes em relação aos valores de referência atuais. Valores de leucócitos e trombócitos tem significado inconclusivo. Hematócrito, hemoglobina, proteína plasmática e valores bioquímicos forneceram importante contribuição para o estabelecimento de parâmetros de referência.(AU)

This paper reports hematological and biochemical values of juveniles Magellanic penguins (Spheniscus magellanicus) stranded in Espírito Santo and Rio de Janeiro, Southeastern Brazil, that were sent for rehabilitation. The average values were: erythrocytes 2.55±0,54 milhões/µL, hemoglobin 13.37±3,91g/dL, hematocrit 41.5±0.04%, plasma protein 6.34±0.81g/dL, leukocytes 16,301±6,402/µL, thrombocytes 20,516±4,591 células/µL3, mean cell volume (MCV) 172.0±53.0 fL, mean cell hemoglobin concentration (MCHC) 26.41±0.02%, alanine transaminase (ALT) 50.0±17.89 U/L, uric acid 8.93±3.0mg/dL, albumin 1.68±0.53mg/dL, calcium 9.7±0.57mg/dL, phosphorus 10.39±8.5mg/dL and glucose 211.6±30.3mg/dL. Values of erythrocytes, MCV and MCH were discrepant in relation to the current reference values. Leukocytes and thrombocytes values has meant inconclusive. Hematocrit, hemoglobin, plasma protein and biochemical values provided important contribution for the establishment of reference parameters.(AU)

Current cariology education in dental schools in Spanish-speaking Latin American countries.

This study sought to provide an overview of current cariology education in Spanish-speaking Latin American dental schools. Data collection was via an eighteen-item survey with questions about curriculum, methods of diagnosis and treatment, and instructors' perceptions about cariology teaching. The response rate was 62.1 percent (n=54), and distribution of participating schools by country was as follows: Bolivia (four), Chile (four), Colombia (twenty-four), Costa Rica (one), Cuba (one), Dominican Republic (two), El Salvador (two), Mexico (six), Panama (two), Peru (four), Puerto Rico (one), Uruguay (two), and Venezuela (one). Forty percent of the responding schools considered cariology the key axis of a course, with a cariology department in 16.7 percent. All schools reported teaching cariology, but with varying hours and at varying times in the curriculum, and 77.8 percent reported having preclinical practices. The majority reported teaching most main teaching topics, except for behavioral sciences, microbiology, saliva and systemic diseases, caries-risk factors, root caries, erosion, and early caries management strategies. The most frequently taught caries detection methods were visual-tactile (96.3 percent), radiographic (92.6 percent), and the International Caries Detection and Assessment System (ICDAS) (61.1 percent). Respondents said their schools' clinics make an operative treatment decision when radiolucency is in the inner half of enamel (42.3 percent) for radiographic criteria and when the lesion is visually non-cavitated (5.8 percent). All respondents reported that their schools teach preventive strategies, but only 43.4 percent said they tie it to risk assessment and 40.7 percent said they implement nonsurgical management regularly.

Role of Cripto-1 during epithelial-to-mesenchymal transition in development and cancer.

Epithelial-to-mesenchymal transition (EMT) is a critical multistep process that converts epithelial cells to more motile and invasive mesenchymal cells, contributing to body patterning and morphogenesis during embryonic development. In addition, both epithelial plasticity and increased motility and invasiveness are essential for the branching morphogenesis that occurs during development of the mammary gland and during tumor formation, allowing cancer cells to escape from the primary tumor. Cripto-1, a member of the epidermal growth factor-Cripto-1/FRL-1/Cryptic (EGF/CFC) gene family, together with the transforming growth factor (TGF)-ß family ligand Nodal, regulates both cell movement and EMT during embryonic development. During postnatal development, Cripto-1 regulates the branching morphogenesis of the mouse mammary gland and enhances both the invasive and migratory properties of mammary epithelial cells in vitro. Furthermore, transgenic mouse models have shown that Cripto-1 promotes the formation of mammary tumors that display properties of EMT, including the down-regulation of the cell surface adherens junctional protein E-cadherin and the up-regulation of mesenchymal markers, such as vimentin, N-cadherin, and Snail. Interestingly, Cripto-1 is enriched in a subpopulation of embryonal, melanoma, prostate, and pancreatic cancer cells that possess stem-like characteristics. Therefore, Cripto-1 may play a role during developmental EMT, and it may also be involved in the reprogramming of differentiated tumor cells into cancer stem cells through the induction of an EMT program.

Alternative splicing enriched cDNA libraries identify breast cancer-associated transcripts.

BACKGROUND: Alternative splicing (AS) is a central mechanism in the generation of genomic complexity and is a major contributor to transcriptome and proteome diversity. Alterations of the splicing process can lead to deregulation of crucial cellular processes and have been associated with a large spectrum of human diseases. Cancer-associated transcripts are potential molecular markers and may contribute to the development of more accurate diagnostic and prognostic methods and also serve as therapeutic targets. Alternative splicing-enriched cDNA libraries have been used to explore the variability generated by alternative splicing. In this study, by combining the use of trapping heteroduplexes and RNA amplification, we developed a powerful approach that enables transcriptome-wide exploration of the AS repertoire for identifying AS variants associated with breast tumor cells modulated by ERBB2 (HER-2/neu) oncogene expression. RESULTS: The human breast cell line (C5.2) and a pool of 5 ERBB2 over-expressing breast tumor samples were used independently for the construction of two AS-enriched libraries. In total, 2,048 partial cDNA sequences were obtained, revealing 214 alternative splicing sequence-enriched tags (ASSETs). A subset with 79 multiple exon ASSETs was compared to public databases and reported 138 different AS events. A high success rate of RT-PCR validation (94.5%) was obtained, and 2 novel AS events were identified. The influence of ERBB2-mediated expression on AS regulation was evaluated by capillary electrophoresis and probe-ligation approaches in two mammary cell lines (Hb4a and C5.2) expressing different levels of ERBB2. The relative expression balance between AS variants from 3 genes was differentially modulated by ERBB2 in this model system. CONCLUSIONS: In this study, we presented a method for exploring AS from any RNA source in a transcriptome-wide format, which can be directly easily adapted to next generation sequencers. We identified AS transcripts that were differently modulated by ERBB2-mediated expression and that can be tested as molecular markers for breast cancer. Such a methodology will be useful for completely deciphering the cancer cell transcriptome diversity resulting from AS and for finding more precise molecular markers.

Epidemiology of HIV and AIDS among adolescents and young adults in the United States.

PURPOSE: To describe the current status of the human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) epidemic among adolescents and young adults in the United States. Despite reported declines in sexual risk behaviors among adolescents in the past decade, little has been published about the epidemiology of HIV and AIDS among adolescents and young adults in the United States. METHODS: We analyzed cases of HIV or AIDS diagnosed among persons aged 13 to 24 years and reported to the national HIV/AIDS Reporting System. We used AIDS cases diagnosed from 1985 through 2003 from the 50 states, the District of Columbia, and the U.S. trusts and territories, and we used HIV cases diagnosed in 2003 from 32 states and the U.S. Virgin Islands. We present five-year trends in HIV diagnoses from 1999 through 2003 from 33 surveillance areas that have stable name-based HIV reporting. The data were adjusted for reporting delays and unreported risk factors. RESULTS: At the end of 2003, 7074 adolescents and young adults, aged 13 to 24 years at the time of diagnosis, were living with AIDS in the United States. Of these, 63% were aged 20 to 24 years. AIDS rates were highest among black persons (63 per 100,000) and youth living in the South (22 per 100,000) and Northeast (18 per 100,000). Among females, the number of diagnosed HIV cases decreased from 1611 cases in 1999 to 1454 in 2003. Among males, the number increased significantly from 1763 in 1999 to 2443 in 2003. The observed increase in the number of HIV diagnoses among males was driven by an increase in HIV diagnoses among young men who have sex with men. CONCLUSIONS: National case surveillance data for persons aged 13 to 24 years revealed that the burden of HIV and AIDS falls most heavily upon the Southern region of the United States and disproportionately upon black and Hispanic youth. The observed increases in the number of HIV cases among men who have sex with men are congruent with recent reports that suggest a resurgence of HIV among these young men. Our findings highlight the need for intensified HIV prevention efforts within minority communities and among men who have sex with men as well as strengthened efforts to encourage at-risk youth to get tested for HIV.

Influenza vaccination trends among adults 65 years or older in the United States, 1989-2002.

BACKGROUND: Influenza vaccination of elderly individuals (65 years or older) has been recommended in the United States since 1961, and consistent surveillance of vaccine use has been conducted since 1989. We examined national trends in influenza vaccination coverage in the United States from 1989 to 2002 among noninstitutionalized elderly individuals and identified factors associated with receipt of influenza vaccine. METHODS: We analyzed data from the 1989-2002 National Health Interview Surveys, weighted to reflect the civilian, noninstitutionalized US population to determine self-reported levels of influenza vaccination. We conducted multivariable logistic regression modeling of 2002 data to identify factors independently associated with self-reported influenza vaccination. RESULTS: Among the elderly, influenza vaccination coverage increased from 30.5% in 1989 to 65.6% in 2002, with only a 2.4% increase from 1997 to 2002. In 2002, coverage remained lower for the non-Hispanic black (49.6%) and Hispanic (48.5%) populations compared with non-Hispanic whites (68.6%). Characteristics associated with a lower likelihood of influenza vaccination included fewer than 4 physician contacts in the past year and whether a person (1) was divorced or separated, (2) was non-Hispanic black or Hispanic, (3) had no regular physician, and (4) had less than a high school education. Individuals with chronic medical conditions and those 75 years or older were more likely to be vaccinated. CONCLUSIONS: By 1997, influenza vaccination coverage exceeded the Healthy People 2000 objective of 60% for the elderly overall, but even by 2002, this objective was still not achieved in the elderly black and Hispanic populations. Vaccination coverage seems to be leveling off, and innovative initiatives are needed to reach the Healthy People 2010 target of 90%, especially among racial and ethnic minorities.

Racial and ethnic disparities in influenza vaccination among elderly adults.

OBJECTIVES: To examine whether access to care factors account for racial/ethnic disparities in influenza vaccination among elderly adults in the United States. DESIGN: Indicators of access to care (predisposing, enabling, environmental/system, and health need) derived from Andersen's behavioral model were identified in the National Health Interview Survey questionnaire. The relationship of these indicators to influenza vaccination and race/ethnicity was assessed with multiple logistic regression models. MAIN RESULTS: Significant differences in vaccination were observed between non-Hispanic (NH) whites (66%) and Hispanics (50%, P<.001) and between NH whites (66%) and NH blacks (46%, P<.001). Controlling for predisposing and enabling access to care indicators, education, marital status, regular source of care, and number of doctor visits, reduced the prevalence odds ratios (POR) comparing Hispanics to non-Hispanic whites from 1.89 to 1.27. For NH blacks, controlling for access to care indicators changed the POR only from 2.24 (95% CI, 1.9 to 2.7) to 1.93 (95% CI, 1.6 to 2.4). CONCLUSIONS: This study confirmed the existence of sizable racial/ethnic differences in influenza vaccination among elderly adults. These disparities were only partially explained by differences in indicators of access to care, especially among non-Hispanic blacks for whom large disparities remained. Factors not available in the National Health Interview Survey, such as patient attitudes and provider performance, should be investigated as possible explanations for the racial/ethnic disparity in influenza vaccination among non-Hispanic blacks.

Antibody response to postexposure prophylaxis in infants born to hepatitis B surface antigen-positive women.

BACKGROUND: Annually 20,000 infants are born to hepatitis B surface antigen (HBsAg)-positive US women. Without prophylaxis 30% risk chronic hepatitis B virus infection, and 25% of those risk dying from resulting liver cirrhosis or liver cancer as adults. METHODS: We attempted to interview each HBsAg-positive pregnant woman reported to the health department between 1992 and 1997, to provide their infants with immunoprophylaxis at birth and in the clinic or home and to serotest at 9 to 15 months of age. RESULTS: Of 879 women reported, 92% enrolled; 787 delivered 796 live infants; 91% of infants received hepatitis B immunoglobulin; 98, 95 and 89% received hepatitis B vaccine (HepB) Doses 1, 2 and 3, respectively; and 80% were serotested. Of these 2.2% were HBsAg-positive and 97% had antibody to HBsAg (anti-HBs) of > or =10 mIU/ml. Anti-HBs concentrations measured in 504 infants were 10 to 99 mIU/ml (25%), 100 to 999 mIU/ml (43%) and > or =1000 mIU/ml (29%). Serotesting was less likely among infants of mothers <20 years of age [odds ratio (OR) 2.5]; white, non-Hispanic (OR 2.8); or with a household income of <$15,000/year (OR 2.0). Lower antibody titers were found when serotesting at 4 to 12 months than at <4 months after HepB-3 (OR 1.8 to 4.4), with HepB-3 receipt <6 months after HepB-2 (OR 2.5) and when household income was <$15,000/year (OR 2.1). CONCLUSIONS: Centralized case management with home visits resulted in high rates of complete immunoprophylaxis and postvaccination testing among infants born to HBsAg-positive women. Perinatal immunoprophylaxis was immunogenic under routine public health use, with higher anti-HBs titers occurring in infants tested <4 months postvaccination. Because infants in households with low income had higher rates of nonprotective antibody responses, they may benefit from extra efforts to ensure that serotesting is conducted postvaccination.