Optical Pretargeting of Tumor with Fluorescent MORF Oligomers

Objective Pretargeting with radioactivity has significantly improved tumor to normal tissue radioactivity ratios over conventional antibody imaging in both animal studies and clinical trials. This laboratory is investigating DNA analogues such as phosphorodiamidate morpholinos (MORFs) for pretargeting using technetium-99m (^99mTc) for detection. However, the unique properties of fluorescence activation and quenching combined with oligomers with their unique properties of hybridization may be particularly useful when used together for pretargeting with optical detection. The use of linear fluorophore-conjugated oligomer duplexes have been little used in animals, and to our knowledge, have not previously been considered for pretargeting applications. Methods A MORF/cDNA pair was selected such that when hybridized, the fluorescence of the Cy5.5-conjugated 25 mer MORF (Cy5.5–MORF25) is inhibited with a BHQ3-conjugated 18 mer complementary DNA (BHQ3–cDNA18). The short BHQ3–cDNA18 was selected to dissociate in the presence of a long cMORF25 in the pretargeted tumor, thus releasing the inhibitor from the Cy5.5 emitter. In this manner, the Cy5.5 fluorescence will be inhibited everywhere but in the target. The dissociation was first examined in vitro by adding the duplex to the cMORF25 both in solution and immobilized on polystyrene microspheres and by surface plasmon resonance (SPR). Thereafter, biotinylated cMORF25 immobilized on streptavidin polystyrene microspheres were administered intramuscularly in one thigh of hairless SKH-1 mice as target while an identical weight of the identical microspheres but without the cMORF25 was administered in the contralateral thigh as control. The animals then received IV the Cy5.5–MORF25/BHQ3–cDNA18 duplex or equal molar dosage of single-chain Cy5.5–MORF25 and were imaged. Results The SPR studies showed that the immobilized cDNA18 rapidly captured the flowing MORF25 to provide a duplex with a slow dissociation rate constant. Furthermore, when cMORF25 was next allowed to flow over the now immobilized duplex, the cDNA18 was unable to prevent dissociation of the heteroduplex and the formation and release of the cMORF25-MORF25 homoduplex. Images of animals obtained soon after receiving the Cy5.5–MORF25 singlet showed intense whole body fluorescence obscuring the target thigh. However, only 5 minutes after receiving the Cy5.5–MORF25/BHQ3–cDNA18 duplex, the target thigh was clearly visible along with only the kidneys. Conclusions This first study of optical pretargeting provides a proof of concept that oligomer pretargeting found to be useful with radioactivity detection is applicable with fluorescent detection as well. In addition, our results demonstrate that by using linear oligomers for optical pretargeting, chain lengths (and base sequences) may be manipulated to provide duplexes with stabilities and fluorescence inhibition optimized for pretargeting and other in vivo applications of optical imaging.     

Life-threatening asthma and anaphylaxis in schools: a treatment model for school-based programs.

BACKGROUND: Pediatric asthma is the No. 1 chronic disease in childhood and is responsible for significant morbidity and mortality. In Nebraska, the number of asthma-related deaths is greater than the national average, and in 1998, 2 students died of acute asthma attacks while attending school in the Omaha public schools (OPSs). In response, we designed and implemented a program to respond to this problem. OBJECTIVE: To implement and study a school-based program for the treatment of life-threatening asthma and anaphylaxis in the OPSs. METHODS: The Emergency Response to Life-Threatening Asthma or Systemic Allergic Reactions (Anaphylaxis) Protocol was designed and evaluated in 78 OPSs from 1998 to 2003. Nurses and school staff were trained in the protocol, which required the use of nebulized albuterol and/or intramuscular epinephrine in conjunction with an emergency response procedure. Outcomes were measured by improvement in acute care in schools and survival of students. Results: In the 5 years of evaluation, 98 students were treated successfully. One student died. Of those treated with the protocol, equal numbers had at school both asthma action plans (AAPs) and metered-dose inhalers (MDIs), MDIs only, or neither AAPs nor MDIs. As a result of the program, there has been an increased awareness from parents, teachers, and physicians about the necessity of an emergency response program. In 2002, an outcome of the OPS program resulted in the formation of Attack on Asthma Nebraska to ensure that Nebraska schools have the education, training, and medications to respond to anyone experiencing a life-threatening asthma or anaphylaxis attack at school. The following year, a revised protocol was approved by the Nebraska State Board of Education for use in all Nebraska schools. CONCLUSIONS: Emergency response protocols provide protection for children while in school. This program should serve as a national model for other school-based programs for children and adolescents with asthma and anaphylaxis.     

Antenatal education – where next?

The history of antenatal education throws light on why contemporary class attenders represent only a particular section of the childbearing population. Since Victorian times, the non-availability of the women's network to middle class women has forced them to seek knowledge of their own bodies, confidence in their childbearing capacities and the support of other women through formal educational opportunities. Research suggests that antenatal classes often fail to provide women with a realistic account of birth and parenting to replace the lived experience of earlier decades and may not be facilitated in such a way as to create the support groups which class attenders critically need. Teaching approaches often promote dependency amongst clients rather than nurturing the decision-making skills required by a consumer-driven maternity service.     

WHO SPEAKS? WHO LISTENS? DIFFERENT VOICES AND DIFFERENT SEXUALITIES

ABSTRACT Psychoanalytic theories of sexuality have regarded homosexuality as a symptom of arrested sexual development. Such theorizing fails to acknowledge the prejudicial values that underpin it. The work of the philosopher, Michel Foucault, can offer psychotherapists new possibilities of considering sexual orientation that do not presuppose a split between the psychic and the socio-political. The complexity and diversity of lesbian roles, identities, experiences and cultures are particularly highlighted by Audre Lorde (a Black lesbian feminist theorist). The two cases illustrate how a Foucauldian analysis, combined with a psychoanalytic approach, can enable us to respond more sensitively to questions of identity in our work with patients.     

High-Dose Naltrexone and Liver Function Safety

Studies have found naltrexone useful in the treatment of diseases other than opiate addiction in which endogenous opioids presumably play a role, such as alcoholism and eating disorders. Some of these studies involve high doses (100–200 mg bid). Because investigational studies with high doses (300 mg/day) reported clinically significant increases in liver enzyme levels, the authors measured a spectrum of liver function parameters in response to high doses of naltrexone in a double-blind, crossover trial (100 mg bid) followed by an open-label period (200 mg bid). They observed no adverse clinical or laboratory changes in liver function in association with high-dose naltrexone therapy in eating disorders.     

Community service utilization by youths hospitalized in a state psychiatric facility

A majority of a cohort of 62 children and adolescents who had been hospitalized in a state psychiatric facility was found to have received less restrictive services such as outpatient mental health services prior to their index admission. Also, a number had been involved with the juvenile justice system and almost two-thirds had been placed out-of-home. Ninety percent had at least one prior psychiatric hospitalization. Just over half of the cohort received case management and individual counseling post release. About a third received family counseling, and a few received other types of services. At least a third were rehospitalized within a year of release. Although 90% of the cohort received some type of service post release, a higher proportion of non service receivers were rehospitalized than service receivers. Even those who received services had a high rate of rehospitalization. These findings raise questions as to the appropriateness of service provision during and following hospitalization.This study was funded by a grant from the Office of Program Evaluation and Research, Ohio Department of Mental Health.     

Rural Demographics Racial/Ethnic Diversification in Metropolitan and Nonmetropolitan Population Change in the United States: Implications for Health Care Provision in Rural America

The diversification of the rural population of the United States provides substantial challenges to the current and to future health care systems in rural areas. Because of a variety of historical, discriminatory, and other factors, minority populations have had lower levels of access to health care in rural as well as urban areas and higher rates of both mortality and morbidity than nonminority populations. Although minority health issues have often been seen as primarily urban issues, this article demonstrates that minority population growth has become a major component of total population growth in rural areas in the past several decades (accounting for nearly 62% of the net growth in the nonmetropolitan population of the United States in the 1980s and for nearly 42% in the 1990s), that future US population growth is likely to be largely a product of minority population growth (nearly 89% of US net population growth from 2000 to 2100 is projected to be due to minority population growth), and that the incidence of diseases and disorders in the US population will come to increasingly involve minority populations (by 2050 roughly 43% of all disease/disorder incidences would involve minority population members). The growth of younger minority populations with disproportionately impoverished socioeconomic characteristics will pose challenges for rural areas and health care systems, which also are likely to face health issues created by disproportionately older populations.