Novel pyrazolo[1,5-a]pyridines with improved aqueous solubility as p110α-selective PI3 kinase inhibitors.

As part of our investigation into pyrazolo[1,5-a]pyridines as novel p110α selective PI3 kinase inhibitors, we report a range of analogues with improved aqueous solubility by the addition of a basic amine. The compounds demonstrated comparable p110α potency and selectivity to earlier compounds but with up to 1000× greater aqueous solubility, as the hydrochloride salts. The compounds also displayed good activity in a cellular assay of PI3 kinase activity.

The Effectiveness of a Physical Activity Educational Campaign in a Rural Obstetrics and Gynecology Office.

Objectives The objective of this study was to evaluate the effectiveness of an early intervention health education campaign to positively influence physical activity (PA) knowledge, intention, and performance among prenatal women and women of reproductive age. Methods This study employed a quantitative, quasi-experimental, control-group comparison design with nonprobability sampling methodology. Implemented in rural healthcare settings located in the Southeastern portion of the United States, participants included prenatal patients and patients of reproductive age (n = 325) from two separate obstetrics and gynecology (OB/GYN) offices. While the intervention group was solicited from an OB/GYN office where the information-based health education campaign was implemented, the comparison group was solicited from a comparable OB/GYN office that did not implement the health education campaign. Results The women exposed to the PA health education campaign were significantly more likely to report that PA information was provided at their physician's office, scored higher on PA knowledge, and were more likely to meet the guidelines for vigorous PA and strength training (p < 0.05). Conclusions Physical activity educational campaigns are a cost effective intervention that can be implemented in healthcare settings to promote maternal and child health.

Exploring the isoform selectivity of TGX-221 related pyrido[1,2-a]pyrimidinone-based Class IA PI 3-kinase inhibitors: synthesis, biological evaluation and molecular modelling.

A novel series of TGX-221 analogues was prepared and tested for their potency against the p110α, p110ß, and p110δ isoforms of the PI3K enzyme, and in two cellular assays. The biological results were interpreted in terms of a p110ß comparative model, in order to account for their selectivity towards this isoform. A CH2NH type linker is proposed to allow binding into the specificity pocket proposed to accommodate the high p110ß-selectivity of TGX-221, although there was limited steric tolerance for substituents on the pendant ring with the 2-position most favourable for substitution.

Perceived barriers to physical activity among pregnant women living in a rural community.

OBJECTIVE: The purpose of this research was to describe perceived barriers to physical activity among pregnant women living in a rural community. DESIGN AND SAMPLE: The project followed a simple descriptive design. The sample included 88 healthy pregnant women from a rural community in the southeast United States. MEASURES: The women completed the International Physical Activity Questionnaire (IPAQ) and an open-ended item seeking a description of personal barriers to engagement in regular physical activity. RESULTS: Scores on the IPAQ were generally equally distributed across categories of low, moderate, and high activity. A total of 42 barriers was described from 34% of the women. Seven themes emerged among the reported barriers: (1) symptoms of pregnancy, (2) family and childrearing activities, (3) lack of personal motivation, (4) time and employment demands, (5) perceptions of sufficient activity from daily life, (6) fear of injury, and (7) lack of a habit of activity. CONCLUSIONS: Barriers reported by the rural women were similar to those identified in other settings. Some perceptions confirmed myths about the health value of exercise during pregnancy, but did not confirm barriers commonly cited or assumed for reduced physical activity among rural residents.

Measuring exercise self-efficacy in pregnant women: psychometric properties of the pregnancy-exercise self-efficacy scale (P-ESES).

BACKGROUND AND PURPOSE: This study assessed the psychometric properties of a modified self-efficacy scale-the Pregnancy-Exercise Self-Efficacy Scale (P-ESES). METHODS: Pregnant women completed the P-ESES and physical activity questionnaires (N = 88). RESULTS: Internal consistency was confirmed by Cronbach's alpha (alpha = 0.838) and equal-length Spearman-Brown (alpha = 8.22). Squared multiple correlation coefficients were calculated showing 9 of 10 items with values greater than the desired .5. A nonrotated exploratory principal components analysis confirmed the same 9 of 10 items loaded on a single factor, accounting for 46.1% of the variance. Each item had an acceptable load value of .40 or higher. CONCLUSIONS: Initial testing of the P-ESES confirmed validity and reliability with the exception of 1 item from the original measure: "Exercising without physician approval".

Therapeutic reactivation of mutant p53 protein by quinazoline derivatives.

PURPOSE: The human tumour suppressor protein p53 is mutated in nearly half of human tumours and most mutant proteins have single amino acid changes. Several drugs including the quinazoline derivative 1 (CP-31398) have been reported to restore p53 activity in mutant cells. The side chain of 1 contains a styryl linkage that compromises its stability and we wished to explore the activity of analogues containing more stable side chains. METHODS: Reactivation of p53 function was measured by flow cytometry as the ability to potentiate radiation-induced G(1)-phase cell cycle arrest and by western blotting to determine expression of p21(WAF1). DNA binding was measured by competition with ethidium and preliminary pharmacological and xenograft studies were carried out. RESULTS: Screening of analogues for potentiation of radiation-induced G(1)-phase cell cycle arrest using NZOV11, an ovarian tumour cell line containing a p53(R248Q) mutation, demonstrated that the (2-benzofuranyl)-quinazoline derivative 5 was among the most active of the analogues. Compound 5 showed similar effects in several other p53 mutant human tumour cell lines but not in a p53 null cell line. 5 also potentiated p21(WAF1) expression induced by radiation. DNA binding affinity was measured and found to correlate with p53 reactivation activity. Plasma concentrations of 5 in mice were sufficient to suggest in vivo activity and a small induced tumour growth delay (7 days) of NZM4 melanoma xenografts was observed. CONCLUSION: Compound 5 restores p53-like function to a human tumour cells lines expressing a variety of mutant p53 proteins, thus providing a basis for the design of further new drugs.

Discovery of pyrazolo[1,5-a]pyridines as p110α-selective PI3 kinase inhibitors.

We have made a novel series of pyrazolo[1,5-a]pyridines as PI3 kinase inhibitors, and demonstrated their selectivity for the p110α isoform over the other Class Ia PI3 kinases. We investigated the SAR around the pyrazolo[1,5-a]pyridine ring system, and found compound 5x to be a particularly potent example (p110α IC(50) 0.9nM). This compound inhibits cell proliferation and phosphorylation of Akt/PKB, a downstream marker of PI3 kinase activity, and showed in vivo activity in an HCT-116 human xenograft model.

Novel pyrazolo[1,5-a]pyridines as p110α-selective PI3 kinase inhibitors: Exploring the benzenesulfonohydrazide SAR.

Structure-activity relationship studies of the pyrazolo[1,5-a]pyridine class of PI3 kinase inhibitors show that substitution off the hydrazone nitrogen and replacement of the sulfonyl both gave a loss of p110α selectivity, with the exception of an N-hydroxyethyl analogue. Limited substitutions were tolerated around the phenyl ring; in particular the 2,5-substitution pattern was important for PI3 kinase activity. The N-hydroxyethyl compound also showed good inhibition of cell proliferation and inhibition of phosphorylation of Akt/PKB, a downstream marker of PI3 kinase activity. It had suitable pharmacokinetics for evaluation in vivo, and showed tumour growth inhibition in two human tumour cell lines in xenograft studies. This work has provided suggestions for the design of more soluble analogues.

Parental perceptions of sibling relationships in families rearing a child with a chronic condition.

This study examined sibling relationships in families raising children with autism, Down syndrome, orthopedic conditions, and diabetes. Parents from 108 families independently completed the 28-item Schaefer Sibling Inventory of Behavior. Parents rated siblings as very empathetic, fairly often kind and involved, and rarely avoidant. Mothers rated sibling empathy higher than fathers did and older siblings more avoidant than younger siblings. Fathers rated male siblings kinder than female siblings; they also rated siblings of children with Down syndrome or autism more kind and involved than siblings of children with orthopedic conditions or diabetes. Sibling intervention efforts should consider these findings and be individualized according to the need of each child and family.

In vivo and in vitro assessment of the action of SN 28049, a benzonaphthyridine derivative targeting topoisomerase II, on the murine Colon 38 carcinoma.

AIM: SN 28049 (N-[2-(dimethylamino)ethyl]-2,6-dimethyl-1-oxo-1,2-dihydrobenzo[b]-1,6-naphthyridine-4-carboxamide) is a new DNA binding drug that targets topoisomerase II. SN 28049 is curative against the murine Colon 38 adenocarcinoma (CT38) while etoposide, another topoisomerase II-directed drug, shows minimal activity; we investigated the basis for this difference in vivo and in vitro. METHODS: Colon 38 tumours were grown in C57Bl mice and in immunodeficient mice. Tumour sections were examined by staining and TUNEL assays. A new cell line (Co-38P) derived from the in vivo tumour was developed and responses were analysed using flow cytometry. RESULTS: Both SN 28049 and etoposide induced similar tumour histological changes, reducing mitotic index and increasing apoptotic index 8 h after administration. At later times however, SN 28049-treated tumours showed further progressive morphological changes while etoposide-treated tumours reverted to their original growth characteristics. The effects of SN 28049 on tumour growth were delayed and attenuated when Colon 38 tumours were grown in immunodeficient mice. SN 28049 and etoposide both induced dose-dependent increases of γ-phosphorylation of histone H2AX and cell cycle perturbation of the Co-38P cell line, indicative of DNA damage, although SN 28049 had 30-fold higher activity. Following 1-hour drug exposure of Co-38P cells, SN 28049 was more effective that etoposide in inducing persistent cycle arrest for the same degree of DNA damage. CONCLUSION: The superior antitumour activity of SN 28049 may result from its ability to induce long term cycle arrest. Host immune responses contribute to the curative activity of SN 28049 and this could result from the induction of cycle arrest.

Action of SN 28049, a new DNA binding topoisomerase II-directed antitumour drug: comparison with doxorubicin and etoposide.

AIM: We have examined the cellular action of SN 28049 (N-[2-(dimethylamino)ethyl]-2,6-dimethyl-1-oxo-1,2-dihydrobenzo[b]-1,6-naphthyridine-4-carboxamide), a DNA binding drug with curative activity against the Colon 38 transplantable murine carcinoma, on human tumour cells. Its action has been compared with that of two topoisomerase II-targetted drugs, etoposide and doxorubicin. METHODS: The NZM3 melanoma and HCT116 colon carcinoma cell lines, each expressing wild-type p53, were cultured and responses were compared by flow cytometry, electrophoresis, microscopy, and growth of tumour xenografts. RESULTS: Responses of NZM3 cells to all three drugs, as measured by histone H2AX γ-phosphorylation, induction of the p53 pathway and cell cycle arrest, were comparable and typical of those of topoisomerase II poisons. Xenografts of NZM3 cells responded to SN 28049 with a tumour growth delay of 16 days. In contrast, HCT116 cells had an attenuated DNA damage response to the drugs and SN 28049 had no in vivo activity, consistent with low topoisomerase II activity. However, SN 28049 inhibited HCT116 cell growth in vitro and activated the p53 pathway to induce a state with G(2)/M-phase DNA content, low mitotic index and a high proportion of binucleate cells. Treated cells expressed cyclin E and the senescence marker ß-galactosidase but showed low expression of cyclin B and survivin. In comparison, etoposide caused little p53 expression or cycle arrest, and doxorubicin had an intermediate effect. CONCLUSION: The action of SN 28049 in NZM3 cells is typical of a topoisomerase II poison, but the low topoisomerase IIα activity of HCT116 cells allowed the detection of a second antiproliferative action of SN 28049 in which cells undergo post-mitotic cycle arrest and induction of p53.

Genomic and signalling pathway characterization of the NZM panel of melanoma cell lines: A valuable model for studying the impact of genetic diversity in melanoma.

Melanoma is a disease associated with a very high mutation burden and thus the possibility of a diverse range of oncogenic mechanisms that allow it to evade therapeutic interventions and the immune system. Here, we describe the characterization of a panel of 102 cell lines from metastatic melanomas (the NZM lines), including using whole-exome and RNA sequencing to analyse genetic variants and gene expression changes in a subset of this panel. Lines possessing all major melanoma genotypes were identified, and hierarchical clustering of gene expression profiles revealed four broad subgroups of cell lines. Immunogenotyping identified a range of HLA haplotypes as well as expression of neoantigens and cancer-testis antigens in the lines. Together, these characteristics make the NZM panel a valuable resource for cell-based, immunological and xenograft studies to better understand the diversity of melanoma biology and the responses of melanoma to therapeutic interventions.

Type 1 diabetes: children and adolescents' knowledge and questions.

AIM: This paper is a report of a study conducted to describe what children and adolescents who have type 1 diabetes know and want to know about the disease. BACKGROUND: Research indicates that young people's knowledge of diabetes may minimize their health complications, because with greater knowledge they may engage in more effective management practices and adherence. METHODS: In this qualitative study, a purposive sample of 58 children and adolescents with type 1 diabetes were interviewed in 2005 about what they knew and wanted to know about their disease. Through a process of induction, major themes were identified from the data. FINDINGS: The six major themes were: (a) Care, including both physical and emotional care, (b) Physiology, (c) Consequences, including both short- and long-term, as well as positive and negative consequences, (d) Cure, (e) Effects on the Family and (f) Experience at Diagnosis. Themes related to the unique challenges associated with type 1 diabetes were also identified. CONCLUSION: Nurses, diabetes educators and parents should provide developmentally appropriate information about diabetes care and management, scaffolding on existing knowledge. They should provide child-centred contexts in which children and adolescents can freely ask questions about their condition and problem-solve. Programmes that allow young people to develop coping skills and share experiences could also prove beneficial.

Synthesis, biological evaluation and molecular modelling of sulfonohydrazides as selective PI3K p110alpha inhibitors.

A series of 2-methyl-5-nitrobenzenesulfonohydrazides were prepared and evaluated as inhibitors of PI3K. An isoquinoline derivative shows good selectivity for the p110alpha isoform over p110beta and p110delta, and also demonstrates good in vitro activity in a cell proliferation assay. Molecular modelling provides a rationalisation for the observed SAR.

Modelling cell population growth with applications to cancer therapy in human tumour cell lines.

In this paper we present an overview of the work undertaken to model a population of cells and the effects of cancer therapy. We began with a theoretical one compartment size structured cell population model and investigated its asymptotic steady size distributions (SSDs) (On a cell growth model for plankton, MMB JIMA 21 (2004) 49). However these size distributions are not similar to the DNA (size) distributions obtained experimentally via the flow cytometric analysis of human tumour cell lines (data obtained from the Auckland Cancer Society Research Centre, New Zealand). In our one compartment model, size was a generic term, but in order to obtain realistic steady size distributions we chose size to be DNA content and devised a multi-compartment mathematical model for the cell division cycle where each compartment corresponds to a distinct phase of the cell cycle (J. Math. Biol. 47 (2003) 295). We then incorporated another compartment describing the possible induction of apoptosis (cell death) from mitosis phase (Modelling cell death in human tumour cell lines exposed to anticancer drug paclitaxel, J. Math. Biol. 2004, in press). This enabled us to compare our model to flow cytometric data of a melanoma cell line where the anticancer drug, paclitaxel, had been added. The model gives a dynamic picture of the effects of paclitaxel on the cell cycle. We hope to use the model to describe the effects of other cancer therapies on a number of different cell lines.

Estimation of radiation-induced interphase cell death in cultures of human tumor material and in cell lines.

A short-term assay method able to estimate the radiation response of human cancer tissue samples would be of great advantage to the individualization of radiotherapy in cancer patients. However, the effect of radiation on [3H]thymidine incorporation by proliferating cells reflects a composite of cell cycle arrest and induced cell death pathways. Here we consider whether it is feasible to correct for cell cycle effects based on comparison of the effects of radiation and the mitotic inhibitor paclitaxel on [3H]thymidine incorporation. Sixty-two short-term (7-day) cultures of human tumor tissue from 61 patients with melanoma, gynecological cancer, brain cancer, and head and neck cancer, as well as 18 5-day cultures of low passage human tumor cell lines, were irradiated at doses from 2 to 9 Gy, or exposed to paclitaxel (200 nM). [3H]Thymidine incorporation was measured at the end of the incubation. Cell cycle times could be estimated from the paclitaxel data and were 2.7 to 18.6 days for melanomas, 2.5 to >40 days for carcinomas, 3.9 to 39 days for brain tumors, and 1.1 to 3.8 days for cell lines. The effects of radiation on [3H]thymidine incorporation varied widely (0-97% and 0-99% inhibition for 2 and 9 Gy, respectively), and in 23 of the clinical samples, but in none of the cell lines, radiation caused significantly greater inhibition of [3H]thymidine incorporation than paclitaxel (p < 0.05). We argue that that these differences reflect radiation-induced cell loss from G1 phase and/or S phase. Responses of short-term cultures of clinical tumor material to radiation, with appropriate correction for cell cycle effects, might have the potential to provide information on radiation-induced cell death in individual patients.

Tobacco use and degenerative joint disease of the spine.

PURPOSE: To examine differences between tobacco users and nonusers who required surgical treatment for degenerative joint disease (DJD) of the spine. DATA SOURCES: Two hundred randomly selected medical records of patients who had undergone surgery for DJD of the spine. CONCLUSIONS: The number of tobacco users in the sample was significantly higher than the number of tobacco users in the general population, indicating greater incidence of DJD among tobacco users. The study demonstrated significant differences between tobacco users and nonusers regarding age, gender, type of occupation, number of imaging studies to diagnosis, and needs for pain management. IMPLICATIONS FOR PRACTICE: Nurse practitioners who deal with education and treatment of patients at risk for spinal degenerative joint disease must consider tobacco use as a significant factor, especially regarding diagnostic studies and pain management.

Comparison of responses of human melanoma cell lines to MEK and BRAF inhibitors.

The NRAS and BRAF genes are frequently mutated in melanoma, suggesting that the NRAS-BRAF-MEK-ERK signaling pathway is an important target for therapy. Two classes of drugs, one targeting activated BRAF and one targeting MEK, are currently undergoing clinical evaluation. We have analysed the NRAS and BRAF mutational status of a series of 44 early passage lines developed from New Zealand patients with metastatic melanoma. 41% of the lines analysed had BRAF mutations, 23% had NRAS mutations, and 36% had neither. We then determined IC50 values (drug concentrations for 50% growth inhibition) for CI-1040, a commonly used inhibitor of MEK kinase; trametinib, a clinical agent targeting MEK kinase; and vemurafenib, an inhibitor of mutant BRAF kinase. Cell lines with activating BRAF mutations were significantly more sensitive to vemurafenib than lines with NRAS mutations or lines lacking either mutation (p < 0.001). IC50 values for CI-1040 and trametinib were strongly correlated (r = 0.98) with trametinib showing ~100-fold greater potency. Cell lines sensitive to vemurafenib were also sensitive to CI-1040 and trametinib, but there was no relationship between IC50 values and NRAS mutation status. A small number of lines lacking a BRAF mutation were sensitive to CI-1040 but resistant to vemurafenib. We used western blotting to investigate the effect on ERK phosphorylation of CI-1040 in four lines, of vemurafenib in two lines and of trametinib in two lines. The results support the view that MEK inhibitors might be combined with BRAF inhibitors in the treatment of melanomas with activated BRAF. The high sensitivity to trametinib of some lines with wildtype BRAF status also suggests that MEK inhibitors could have a therapeutic effect against some melanomas as single agents.

Costs of treating depression with individual versus family therapy.

Depression is one of the most common concerns that bring clients to treatment. Although marriage and family therapy has been shown to be an effective treatment, little research exists regarding the cost-effectiveness of related services. In this study, we examined claims data for 164,667 individuals diagnosed with depression to determine (a) differences in the cost of treating depression according to type of therapy and license type, (b) differences in recidivism rates by age, gender, type of therapy, and type of mental health professional, and (c) differences in cost-effectiveness by therapy modality and type of professional. The results showed that services provided by marriage and family therapists resulted in the lowest recidivism rate, and family therapy services were the least expensive.

Assessment of smokeless tobacco use in the history and physical examination by primary healthcare providers.

PURPOSE: Following a simple descriptive research design, we examined how and to what extent primary healthcare providers in rural southern regions of the United States ask patients about the use of smokeless tobacco as indicated in the document used for the patient history. DATA SOURCES: Copies of blank history and physical forms used in offices of primary care providers in Alabama, Georgia, South Carolina, and Tennessee were examined to identify items related specifically to tobacco use. CONCLUSIONS: Twenty-nine providers returned history and physical forms, which revealed 24% showed no item related to tobacco use. Others included questions related to smoking, but only 7% mentioned any sort of smokeless tobacco use. IMPLICATIONS FOR PRACTICE: Although a few studies have suggested the use of smokeless tobacco to be less harmful than smoking, all forms of smokeless tobacco are recognized carcinogens and dangerous for health. It is not sufficient to simply ask patients about smoking behaviors. Primary care providers, especially nurse practitioners, have the unique opportunity to assess use of smokeless tobacco and to offer treatment and counsel to help patients to stop the behavior.