Developing a Web-Based Course in Phases

Introduction: Distance education has grown in popularity and usage. At the same time, enrollments at postsecondary institutions continue to increase. This places significant growth pressures on institutions of higher learning. Institutions providing nursing education have historically faced limited faculty resources. This has made it difficult to meet demand for distance education; and an “all-at-once” approach to course development does little to ease this problem. In this approach, resources are expended up front before a course is offered. [See PDF for complete abstract].

Therapeutic Targeting of ATP7B in Ovarian Carcinoma.

PURPOSE: Resistance to platinum chemotherapy remains a significant problem in ovarian carcinoma. Here, we examined the biological mechanisms and therapeutic potential of targeting a critical platinum resistance gene, ATP7B, using both in vitro and in vivo models. EXPERIMENTAL DESIGN: Expression of ATP7A and ATP7B was examined in ovarian cancer cell lines by real-time reverse transcription-PCR and Western blot analysis. ATP7A and ATP7B gene silencing was achieved with targeted small interfering RNA (siRNA) and its effects on cell viability and DNA adduct formation were examined. For in vivo therapy experiments, siRNA was incorporated into the neutral nanoliposome 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC). RESULTS: ATP7A and ATP7B genes were expressed at higher levels in platinum-resistant cells compared with sensitive cells; however, only differences in ATP7B reached statistical significance. ATP7A gene silencing had no significant effect on the sensitivity of resistant cells to cisplatin, but ATP7B silencing resulted in 2.5-fold reduction of cisplatin IC(50) levels and increased DNA adduct formation in cisplatin-resistant cells (A2780-CP20 and RMG2). Cisplatin was found to bind to the NH(2)-terminal copper-binding domain of ATP7B, which might be a contributing factor to cisplatin resistance. For in vivo therapy experiments, ATP7B siRNA was incorporated into DOPC and was highly effective in reducing tumor growth in combination with cisplatin (70-88% reduction in both models compared with controls). This reduction in tumor growth was accompanied by reduced proliferation, increased tumor cell apoptosis, and reduced angiogenesis. CONCLUSION: These data provide a new understanding of cisplatin resistance in cancer cells and may have implications for therapeutic reversal of drug resistance.

Multiple GABA receptors in rabbit retina

The task of encoding and processing complex sensory input requires many types of transsynaptic signals. This requirement is served in part by an extensive group of neurotransmitter substances which may include thirty or more different compounds. At the next level of information processing, the existence of multiple receptors for a given neurotransmitter appears to be a widely used mechanism to generate multiple responses to a given first messenger (Snyder and Goodman, 1980). Despite the wealth of published data on GABA receptors, the existence of more than one GABA receptor was in doubt until the mid 1980's. Presently there is still disagreement on the number of types of GABA receptors, estimates for which range from two to four (DeFeudis, 1983; Johnston, 1985). Part of the problem in evaluating data concerning multiple receptor types is the lack of information on the number of gene products and their subsequent supramolecular organization in different neurons. In order to evaluate the question concerning the diversity of GABA receptors in the nervous system, we must rely on indirect information derived from a wide variety of experimental techniques. These include pharmacological binding studies to membrane fractions, electrophysiological studies, localization studies, purification studies, and functional assays. Almost all parts of the central and peripheral nervous system use GABA as a neurotransmitter, and these experimental techniques have therefore been applied to many different parts of the nervous system for the analysis of GABA receptor characteristics. We are left with a large amount of data from a wide variety of techniques derived from many parts of the nervous system. When this project was initiated in 1983, there were only a handful of pharmacological tools to assess the question of multiple GABA receptors. The approach adopted was to focus on a single model system, using a variety of experimental techniques, in order to evaluate the existence of multiple forms of GABA receptors. Using the in vitro rabbit retina, a combination of pharmacological binding studies, functional release studies and partial purification studies were undertaken to examine the GABA receptor composition of this tissue. Three types of GABA receptors were observed: Al receptors coupled to benzodiazepine and barbiturate modulation, and A2 or uncoupled GABA-A receptors, and GABA-B receptors. These results are evaluated and discussed in light of recent findings by others concerning the number and subtypes of GABA receptors in the nervous system. ^

Cultural beliefs and hypertension in rural Philippines

Hypertension (HTN), the major risk factor for cardiovascular disease (CVD), is emerging as a major public health problem in the Philippines. CVD has been the leading cause of mortality in the Philippines since 1990. ^ Although research has shown that certain populations have a greater propensity for HTN, and that culture may be a factor, empirical investigations of the influence of cultural beliefs on HTN are lacking. ^ The operational aims of this study were to: (a) develop and examine the reliability (test-retest, internal consistency) and validity (content) of a questionnaire which measures factors related to HTN; (b) administer the questionnaire; and (c) measure blood pressure, height, and weight of the ≥ 30 year old residents of San Antonio, Nueva Ecija, Philippines. ^ The analytic aims were to determine the: (a) cultural beliefs relating to HTN; (b) associations between cultural beliefs and HTN; and (c) extent to which cultural beliefs versus biological, behavioral, socioeconomic, and access factors are associated with HTN. ^ A cluster survey was conducted among 336 residents ≥ 30 years old in May, 1998. Sixty clusters of households were derived using probability proportionate to size sampling technique. Seven households per cluster were visited and one respondent per household was randomly chosen for interview and measurement of blood pressure, height and weight. A response rate of 84% (336/400) was achieved. ^ Results showed that the test-retest reliability of cultural belief items was 0.69–0.96. Internal consistency reliability was 0.74. ^ HTN (SBP ≥ 140; or DBP ≥ 90 mmHg; or currently taking anti-hypertensive medication) prevalence was 23/100. Univariate logistic regression showed cultural beliefs to be significantly associated (p < 0.037) with HTN. However, multivariate analysis showed that only age ≥ 50 (p = 0.000), family history of HTN (p = 0.004) and body mass index ≥ 25 (p = 0.003) were significant predictors. ^ In the absence of fully implemented programs to prevent and control HTN, the current prevalence is only expected to increase, leading to substantial increases in morbidity and mortality and health care cost. It is recommended that research which focuses on designing, implementing, and evaluating culturally appropriate community-wide programs on HTN prevention and control be undertaken in this community. ^

PHENOTYPIC ASSOCIATION OF GENE AMPLIFICATION WITH MULTIPLE DRUG RESISTANCE IN VINCRISTINE RESISTANT CHINESE HAMSTER OVARY CELLS

Resistance of tumors to pharmacologic agents poses a significant problem in the treatment of human malignancies. This study overviews the scope of clinical resistance and focuses upon current research attempts toward investigation of the phenomenon of multidrug resistance (MDR).^ The objective of this investigation was to determine whether gene amplification had a role in the development of the MDR phenotype in Chinese hamster ovary cells (CHO) primarily selected for resistance to vincristine (VCR). A DNA fragment, previously shown to be amplified in two independently derived Chinese hamster cell lines exhibiting the MDR phenotype, was also amplified in VCR hamster lines. Sequences flanking this fragment were shown to contain coding information for a 4.3 kb transcript overproduced in VCR cells. These sequences were not enriched in double minute DNA preparations isolated from VCR cells. There was an approximately forty-fold increase in both the level of gene amplification and transcript overproduction in the VCR cell lines, independent of the level of primary resistance. This DNA amplification and overproduction of the 4.3 kb transcript was also demonstrated in CHO cells independently selected for resistance to Adriamycin and vinblastine.^ All the DNA sequences of two hamster cDNA clones containing 785 and 932 base pair inserts showed direct homology to the published mouse mdr sequences (about 90%). This sequence conservation held for only portions of the gene when the human mdr1 sequences were compared with those from either the mouse or hamster.^ Somatic cell hybrids, constructed between VCR CHO cells and sensitive murine cells, were used to determine whether there was a functional relationship between the chromosome bearing the amplified sequences and the MDR phenotype. Concordant segregation between vincristine resistance, the MDR phenotype, the presence of MDR-associated amplified sequences, overexpression of the mRNA encoded by these sequences, overexpression of the mRNA encoded by these sequences, and CHO chromosome Z1 was consistent with the hypothesis that there is an amplified gene on chromosome Z1 of the VCR CHO cells which is responsible for MDR in these cells. ^