Development of a surface-enhanced raman spectroscopy method for the detection of benzodiazepines in urine

Benzodiazepines are among the most prescribed compounds for anti-anxiety and are present in many toxicological screens. These drugs are also prominent in the commission of drug facilitated sexual assaults due their effects on the central nervous system. Due to their potency, a low dose of these compounds is often administered to victims; therefore, the target detection limit for these compounds in biological samples is 10 ng/mL. Currently these compounds are predominantly analyzed using immunoassay techniques; however more specific screening methods are needed. ^ The goal of this dissertation was to develop a rapid, specific screening technique for benzodiazepines in urine samples utilizing surface-enhanced Raman spectroscopy (SERS), which has previously been shown be capable of to detect trace quantities of pharmaceutical compounds in aqueous solutions. Surface enhanced Raman spectroscopy has the advantage of overcoming the low sensitivity and fluorescence effects seen with conventional Raman spectroscopy. The spectra are obtained by applying an analyte onto a SERS-active metal substrate such as colloidal metal particles. SERS signals can be further increased with the addition of aggregate solutions. These agents cause the nanoparticles to amass and form hot-spots which increase the signal intensity. ^ In this work, the colloidal particles are spherical gold nanoparticles in aqueous solution with an average size of approximately 30 nm. The optimum aggregating agent for the detection of benzodiazepines was determined to be 16.7 mM MgCl2, providing the highest signal intensities at the lowest drug concentrations with limits of detection between 0.5 and 127 ng/mL. A supported liquid extraction technique was utilized as a rapid clean extraction for benzodiazepines from urine at a pH of 5.0, allowing for clean extraction with limits of detection between 6 and 640 ng/mL. It was shown that at this pH other drugs that are prevalent in urine samples can be removed providing the selective detection of the benzodiazepine of interest. ^ This technique has been shown to provide rapid (less than twenty minutes), sensitive, and specific detection of benzodiazepines at low concentrations in urine. It provides the forensic community with a sensitive and specific screening technique for the detection of benzodiazepines in drug facilitated assault cases.^

PERSONALIDADE, ESTILOS PARENTAIS E COGNIÇÃO EM MENINOS VÍTIMAS DE ABUSO SEXUAL

Há um consenso na literatura de que a vivência do abuso sexual na infância ocasiona uma série de consequências, tanto físicas quanto psicológicas, às vítimas. O cenário de violência contra crianças e adolescentes está cada vez mais frequente na sociedade contemporânea, tornando-se um problema de saúde pública, de modo a ser necessário um olhar mais atento dos profissionais e elaboração de políticas públicas tanto para a prevenção dos casos como para o tratamento dos problemas deles decorrentes. Nas crianças, há uma associação do aumento de problemas comportamentais e psicológicos, que podem se estender até o período da idade adulta. Outros estudos enfatizam alterações em áreas cerebrais e as relacionam com o surgimento de transtornos mentais na população de crianças vítimas de violência sexual, dentre outros tipos de violência. O objetivo deste trabalho foi verificar a presença de traços de personalidade, os estilos parentais desenvolvidos e o desempenho cognitivo, especificamente as funções executivas, em meninos com histórico de abuso sexual. Foram avaliadas 62 crianças e adolescentes (32 com histórico de abuso sexual e 30 sem), sendo que no grupo de abuso a idade média dos integrantes é de 11,7 anos de idade e seus genitores majoritariamente divorciados, diferentemente do grupo controle, no qual os genitores são, na maior parte, casados. O método constou de questionário sociodemográfico, testes neuropsicológicos que avaliam as funções executivas (Wisconsin Card Sortint Test, Figuras de Ray e Escala de Inteligência Wechsler para Crianças - Terceira Edição (WISC-III)- Semelhanças, Arranjo de Figuras, Compreensão, Cubos, Dígitos e Vocabulário), Questionário de Personalidade para Crianças e Adolescentes (EPQ-J) e Inventário de Estilos Parentais (IEP). Não houve resultados significativos entre o abuso e as funções executivas, tanto nos subtestes dos Escala de Inteligência Wechsler para Crianças - Terceira Edição (WISC-III) quanto no do Wisconsin; porém o grupo apresentou dados significativamente menores na memória do Figuras Complexas de Rey. Com relação ao EPQ-J, os resultados demonstraram maiores índices de neuroticismo, os quais podem ser associados com sintomas de depressão, e apresentaram menores índices em extroversão, trazendo possíveis dificuldades nas relações sociais. Com relação aos estilos parentais, o grupo apresentou dificuldades e prejuízos nas percepções sobre os genitores, especialmente com relação ao genitor, que é o maior abusador estatisticamente. Os dados mostraram uma correlação entre neuroticismo e vocabulário, estilos parentais e vocabulário, e neuroticismo e estilos parentais. Observou-se a importância de maiores estudos com o público masculino, para que possam ser elucidadas, de forma mais abrangente, as questões de abuso sexual e suas interferências nos aspectos neuropsicológicos, estilos parentais e traços de personalidade, visto que a maior parte dos estudos é relacionada ao público feminino. É necessário o desenvolvimento de políticas públicas para o atendimento específico do público masculino que sofre abuso, e que o profissional da saúde esteja preparado para ajudar, fortalecendo os aspectos psíquicos e emocionais.

Placental contribution to the origins of sexual dimorphism in health and diseases: sex chromosomes and epigenetics

Sex differences occur in most non-communicable diseases, including metabolic diseases, hypertension, cardiovascular disease, psychiatric and neurological disorders and cancer. In many cases, the susceptibility to these diseases begins early in development. The observed differences between the sexes may result from genetic and hormonal differences and from differences in responses to and interactions with environmental factors, including infection, diet, drugs and stress. The placenta plays a key role in fetal growth and development and, as such, affects the fetal programming underlying subsequent adult health and accounts, in part for the developmental origin of health and disease (DOHaD). There is accumulating evidence to demonstrate the sex-specific relationships between diverse environmental influences on placental functions and the risk of disease later in life. As one of the few tissues easily collectable in humans, this organ may therefore be seen as an ideal system for studying how male and female placenta sense nutritional and other stresses, such as endocrine disruptors. Sex-specific regulatory pathways controlling sexually dimorphic characteristics in the various organs and the consequences of lifelong differences in sex hormone expression largely account for such responses. However, sex-specific changes in epigenetic marks are generated early after fertilization, thus before adrenal and gonad differentiation in the absence of sex hormones and in response to environmental conditions. Given the abundance of X-linked genes involved in placentogenesis, and the early unequal gene expression by the sex chromosomes between males and females, the role of X- and Y-chromosome-linked genes, and especially those involved in the peculiar placenta-specific epigenetics processes, giving rise to the unusual placenta epigenetic landscapes deserve particular attention. However, even with recent developments in this field, we still know little about the mechanisms underlying the early sex-specific epigenetic marks resulting in sex-biased gene expression of pathways and networks. As a critical messenger between the maternal environment and the fetus, the placenta may play a key role not only in buffering environmental effects transmitted by the mother but also in expressing and modulating effects due to preconceptional exposure of both the mother and the father to stressful conditions.

Neural Mechanisms of Young Adult Sexual Decision-Making and Risk Behavior

Sexual risk behavior among young adults is a serious public health concern; 50% will contract a sexually transmitted infection (STI) before the age of 25. The current study collected self-report personality and sexual history data, as well as neuroimaging, experimental behavioral (e.g., real-time hypothetical sexual decision making data), and self-report sexual arousal data from 120 heterosexual young adults ages 18-26. In addition, longitudinal changes in self-reported sexual behavior were collected from a subset (n = 70) of the participants. The primary aims of the study were (1) to predict differences in self-report sexual behavior and hypothetical sexual decision-making (in response to sexually explicit audio-visual cues) as a function of ventral striatum (VS) and amygdala activity, (2) test whether the association between sexual behavior/decision-making and brain function is moderated by gender, self-reported sexual arousal, and/or trait-level personality factors (i.e., self-control, impulsivity, and sensation seeking) and (3) to examine how the main effects of neural function and interaction effects predict sexual risk behavior over time. Our hypotheses were mostly supported across the sexual behavior and decision-making outcome variables, such that neural risk phenotypes (heightened reward-related ventral striatum activity coupled with decreased threat-related amygdala activity) were associated with greater lifetime sexual partners at baseline measured and over time (longitudinal analyses). Impulsivity moderated the relationship between neural function and self-reported number of sexual partners at baseline and follow up measures, as well as experimental condom use decision-making. Sexual arousal and sensation seeking moderated the relationship between neural function and baseline and follow up self-reports of number of sexual partners. Finally, unique gender differences were observed in the relationship between threat and reward-related neural reactivity and self-reported sexual risk behavior. The results of this study provide initial evidence for the potential role for neurobiological approaches to understanding sexual decision-making and risk behavior. With continued research, establishing biomarkers for sexual risk behavior could help inform the development of novel and more effective individually tailored sexual health prevention and intervention efforts.

Development of a Surface-Enhanced Raman Spectroscopy Method for the Detection of Benzodiazepines in Urine

Benzodiazepines are among the most prescribed compounds for anti-anxiety and are present in many toxicological screens. These drugs are also prominent in the commission of drug facilitated sexual assaults due their effects on the central nervous system. Due to their potency, a low dose of these compounds is often administered to victims; therefore, the target detection limit for these compounds in biological samples is 10 ng/mL. Currently these compounds are predominantly analyzed using immunoassay techniques; however more specific screening methods are needed. The goal of this dissertation was to develop a rapid, specific screening technique for benzodiazepines in urine samples utilizing surface-enhanced Raman spectroscopy (SERS), which has previously been shown be capable of to detect trace quantities of pharmaceutical compounds in aqueous solutions. Surface enhanced Raman spectroscopy has the advantage of overcoming the low sensitivity and fluorescence effects seen with conventional Raman spectroscopy. The spectra are obtained by applying an analyte onto a SERS-active metal substrate such as colloidal metal particles. SERS signals can be further increased with the addition of aggregate solutions. These agents cause the nanoparticles to amass and form hot-spots which increase the signal intensity. In this work, the colloidal particles are spherical gold nanoparticles in aqueous solution with an average size of approximately 30 nm. The optimum aggregating agent for the detection of benzodiazepines was determined to be 16.7 mM MgCl2, providing the highest signal intensities at the lowest drug concentrations with limits of detection between 0.5 and 127 ng/mL. A supported liquid extraction technique was utilized as a rapid clean extraction for benzodiazepines from urine at a pH of 5.0, allowing for clean extraction with limits of detection between 6 and 640 ng/mL. It was shown that at this pH other drugs that are prevalent in urine samples can be removed providing the selective detection of the benzodiazepine of interest. This technique has been shown to provide rapid (less than twenty minutes), sensitive, and specific detection of benzodiazepines at low concentrations in urine. It provides the forensic community with a sensitive and specific screening technique for the detection of benzodiazepines in drug facilitated assault cases.

Suppression of the Insulin Receptors in Adult Schistosoma japonicum Impacts on Parasite Growth and Development: Further Evidence of Vaccine Potential

To further investigate the importance of insulin signaling in the growth, development, sexual maturation and egg production of adult schistosomes, we have focused attention on the insulin receptors (SjIRs) of Schistosoma japonicum, which we have previously cloned and partially characterised. We now show, by Biolayer Interferometry, that human insulin can bind the L1 subdomain (insulin binding domain) of recombinant (r)SjIR1 and rSjIR2 (designated SjLD1 and SjLD2) produced using the Drosophila S2 protein expression system. We have then used RNA interference (RNAi) to knock down the expression of the SjIRs in adult S. japonicum in vitro and show that, in addition to their reduced transcription, the transcript levels of other important downstream genes within the insulin pathway, associated with glucose metabolism and schistosome fecundity, were also impacted substantially. Further, a significant decrease in glucose uptake was observed in the SjIR-knockdown worms compared with luciferase controls. In vaccine/challenge experiments, we found that rSjLD1 and rSjLD2 depressed female growth, intestinal granuloma density and faecal egg production in S. japonicum in mice presented with a low dose challenge infection. These data re-emphasize the potential of the SjIRs as veterinary transmission blocking vaccine candidates against zoonotic schistosomiasis japonica in China and the Philippines.

Machinima na educação sexual: intervenção em meio escolar

Relatório de estágio apresentado para a obtenção do grau de mestre na área de Educação e Comunicação Multimédia

Machinima na educação sexual: intervenção em meio escolar

Relatório de estágio apresentado para a obtenção do grau de mestre na área de Educação e Comunicação Multimédia

Development of water-soluble polyanionic carbosilane dendrimers as novel and highly potent topical anti-HIV-2 microbicides

This is the author’s version of a work that was accepted for publication in Nanoscale.

Sexual dysfunction in breast cancer survivors: cross-cultural adaptation of the Sexual Activity Questionnaire for use in Portugal

"Introduction: The increasing survivor population of breast cancer has shifted research and practice interests into the impacts of the disease and treatment in quality of life aspects. The lack of tools available in Portuguese to objectively evaluate sexual function led to the development of this study, which aimed to cross-culturally adapt and validate the Sexual Activity Questionnaire for use in Portugal. Material and Methods: The questionnaire was translated and back-translated, refined following face-to-face interviews with seven breast cancer survivors, and then self-administered by a larger sample at baseline and a fortnight later to test validity and reliability. Results: Following cognitive debriefing (n = 7), minor changes were made and the Sexual Activity Questionnaire was then tested with 134 breast cancer survivors. A 3-factor structure explained 75.5% of the variance, comprising the Pleasure, Habit and Discomfort scales, all yielding good internal consistency (Cronbach’s α > 0.70). Concurrent validity with the FACt-An and the BCPT checklist was good (Spearman’s r > 0.65; p-value < 0.001) and reliability acceptable (Cohen’s k > 0.444). The Sexual Activity Questionnaire allowed the identification of 23.9% of sexually inactive women, for whom the main reasons were lack of interest or motivation and not having a partner. Discussion: Patient-reported outcomes led to a more comprehensive and improved approach to cancer, tackling areas previously abandoned. Future research should focus on the validation of this scale in samples with different characteristics and even in the overall population to enable generalizability of the findings. Conclusion: The adapted Sexual Activity Questionnaire is a valid tool for assessing sexual function in breast cancer survivors in Portugal."

Comportamiento sexual y reproductivo de los estudiantes de la carrera de Medicina, Facultad de Ciencias Médicas, Universidad de Cuenca, en el año 2015, Cuenca

La salud sexual y reproductiva es fundamental en el desarrollo biológico, psicológico y social del ser humano, el desconocimiento y la falta de información ponen en riesgo el correcto desenvolvimiento del ser humano en la sociedad, limita sus aspiraciones y metas y favorece la desestructuración familiar. OBJETIVO: Identificar el comportamiento sexual y reproductivo de los estudiantes de la carrera de medicina de la Universidad de Cuenca, para proporcionar una realidad y analizar posibles soluciones respecto a los resultados obtenidos. MATERIALES Y MÉTODOS: Es un estudio descriptivo en el año 2015. La muestra se obtuvo mediante aleatorización por conglomerados, constituido por hombres y mujeres. Para obtener la información se aplicó un cuestionario. La muestra estuvo conformada por 179 estudiantes, el nivel de confianza del 95% y un error estándar de 5%. Los datos fueron ingresados y analizados en el programa estadístico Epi Info 7. RESULTADOS: La población estudiantil con un 24,14% inició su vida sexual a los 18 años. El 48,6% refirió haber tenido relaciones sexuales. El 82,76% afirma haber utilizado un método anticonceptivo. CONCLUSIONES: Se evidencia que a pesar de disponer de recursos para acceder a servicios de salud, tener conocimientos sobre prevención de embarazo y que el uso de anticoncepción es frecuente, existen todavía estudiantes que necesitan capacitación en esta temática

Reproduction, egg morphology and development observed in Monographis queenslandicus (Diplopoda: Polyxenidae)

This study provides new insight into penicillate sexual behaviour and eggdevelopment as observed in Monographis queenslandicus Huynh et Veenstra, 2013 (Polyxenidae). The developing eggs were found to have two distinct stages, namely the chorion and pupoid, which both proved to be of 12–14 days duration. Both stages were characterized by distinctive external morphology. Morphological features observed pre ecdysis included the development of a smooth, tough membrane of the chorion. In contrast, the pupoid stage exhibited an embryonic cuticle with tiny spines, which were later being used to rupture the chorion. Additionally, an aperture bordered by protective sensilla located on the anterior of the pupoid is described for the first time.How to cite this article: Huynh C., Veenstra A.A. 2014. Reproduction, egg morphology and development observed in Monographis queenslandicus (Diplopoda: Polyxenidae)

Early-life sexual segregation: ontogeny of isotopic niche differentiation in the Antarctic fur seal

Investigating the ontogeny of niche differentiation enables to determine at which life-stages sexual segregation arises, providing insights into the main factors driving resource partitioning. We investigated the ontogeny of foraging ecology in Antarctic fur seals (Arctocephalus gazella), a highly dimorphic species with contrasting breeding strategies between sexes. Sequential δ(13)C and δ(15)N values of whiskers provided a longitudinal proxy of the foraging niche throughout the whole life of seals, from weaning, when size dimorphism is minimal to the age of 5. Females exhibited an early-life ontogenetic shift, from a total segregation during their first year at-sea, to a similar isotopic niche as breeding females as early as age 2. In contrast, males showed a progressive change in isotopic niche throughout their development such that 5-year-old males did not share the same niche as territorial bulls. Interestingly, males and females segregated straight after weaning with males appearing to feed in more southerly habitats than females. This spatial segregation was of similar amplitude as observed in breeding adults and was maintained throughout development. Such early-life niche differentiation is an unusual pattern and indicates size dimorphism and breeding constraints do not directly drive sexual segregation contrary to what has been assumed in otariid seals.

Predictors of Adolescent Sexual Intentions and Behavior: Attitudes, Parenting, and Neighborhood Risk

The current study was a cross-sectional examination of data collected during an HIV risk reduction intervention in south Florida. The purpose of the study was to explore the relationships between neighborhood stress, parenting, attitudes, and adolescent sexual intentions and behavior. The Theory of Planned Behavior was used as a model to guide variable selection and propose an interaction pathway between predictors and outcomes. Potential predictor variables measured for adolescents ages 13-18 (n=196) included communication about sex, parent-family connectedness, parental presence, parent-adolescent activity participation, attitudes about sex and condom use, neighborhood disorder, and exposure to violence. Outcomes were behavioral intentions and sexual behavior for the previous eight months. Neighborhood data was supplemented with ZIP Code level data from regional sources and included median household income, percentage of minority and Hispanic residents, and number of foreclosures. Statistical tests included t-tests, Pearson’s correlations, and hierarchical linear regressions. Results showed that males and older adolescents reported less positive behavioral intentions than females and adolescents younger than 16. Intentions were associated with condom attitudes, sexual attitudes, and parental presence; unprotected sexual behavior was associated with parental presence. The best fit model for intentions included gender, sexual attitudes, condom attitudes, parental presence, and neighborhood disorder. The unsafe sexual behavior model included whether the participant lived with both natural parents in the previous year, and the percent of Hispanic residents in the neighborhood. Study findings indicate that more research on adolescent sexual behavior is warranted, specifically examining the differentials between variables that affect intentions and those that affect behavior. A focus on gender and age differences during intervention development may allow for better targeting and more efficacious interventions. Adding peer and media influences to the framework of attitudes, parenting, and neighborhood may offer more insight into patterns of adolescent sexual behavior risk.

Sex-Positive Curricula: An Investigation of the Relationship Between Physical Fitness, Self-Concept and Sexual Functioning

Despite the well-recognized benefits of exercise, Americans are gaining weight in astounding proportions and levels of physical activity are on the decline. The purpose of this study was to investigate a relationship between physical fitness, self-concept and sexual health. There is a dearth of knowledge on this relationship specifically in the context of sex-negative curricula, which is the dominate discourse in the United States. One hundred and thirty-three participants between the ages of 18 - 50 volunteered for fitness testing and data collection. Physical fitness was assessed through body fat, resting metabolic rate, cardiovascular endurance, muscular strength, muscular endurance and flexibility. Self-reported exercise was measured using the International Physical Activity Questionnaire. Self-concept was measured by the Six Factor Self-Concept Scale, which presented a total self-concept score and as six individual concepts of self (likability, morality, task accomplishment, giftedness, power and vulnerability). Additionally, sexual function was measured by Derogatis Interview for Sexual Functioning and presented as both an aggregate score and five separate constructs of sexual functioning (fantasy/cognition, arousal, orgasm, behavior/experience, and drive/desire). Questions pertaining to sexual partners, sex education, and demographic information were also included. The results of the General Linear Model indicated significant relationships between physical fitness, self-concept and total sexual functioning. The sexual behavior/experience of men was predicted by body fat percentage and flexibility. In women, behavior/experience was predicted by body fat percentage and arousal was predicted by cardiovascular endurance. Total self-concept was related to muscular endurance. When men were isolated in the analysis, likability was positively related to sexual behavior/experience, and task accomplishment was inversely related to sexual behavior/experience. In women, giftedness was related to cognition/fantasy, arousal, orgasm and total sexual functioning. No relationships were found between physical fitness and the number of sexual partners in men; however, both muscular strength and the power self-concept were significantly related to number of sexual partners in women. As a result of these findings, women may be inclined to exercise to improve arousal and sexual functioning. Furthermore, educators should note the findings of a positive relationship between physical and psychological health and sexual well-being because they provide support for the development and adoption of sex-positive curricula that incorporate potential benefits of sexual activity.