Enhanced Retroviral Transduction of 5-Fluorouracil-Resistant Human Bone Marrow (Stem) Cells Using a Genetically Modified Packaging Cell Line

Pluripotent hematopoietic stem cells (PHSC) are rare cells capableof multilineage differentiation, long-term reconstituting activity andextensive self-renewal. Such cells are the logical targets for manyforms of corrective gene therapy, but are poor targets for retroviralmediated gene transfer owing to their quiescence, as retroviraltransduction requires that the target cells be cycling. To try andsurmount this problem we have constructed a retroviral producer linethat expresses the membrane-bound form of human stem cell factor (SCF)on its cell surface. These cells are capable, therefore, of deliveringa growth signal concomitant with recombinant retroviral vectorparticles. In this report we describe the use of this cell line totransduce a highly quiescent population of cells isolated from adulthuman bone marrow using the 5-fluorouracil (FU) resistance technique ofBerardi et al. Quiescent cells selected using this technique weretransduced by cocultivation with retroviral producers expressingsurface bound SCF or with the parent cell line that does not. Followingcoculture, the cells were plated in long-term bone marrow culture for afurther 5 weeks, before plating the nonadherent cells in semisolidmedia. Colonies forming in the semisolid media over the next 14 dayswere analyzed by polymerase chain reaction for the presence of theretroviral vector genome. Over six experiments, the transductionfrequency of the quiescent 5-FU resistant cells using theSCF-expressing producer line averaged about 20%, whereas thosetransduced using the parent producer line showed evidence of reducedlevels or no transduction.     

Psychometric evaluation of a patient-reported outcome measure in pancreatic exocrine insufficiency (PEI)

Background/objectives

Pancreatic exocrine insufficiency (PEI) is commonly caused by chronic pancreatitis (CP) or cystic fibrosis (CF). There are no PEI-specific patient-reported assessments of symptoms and impacts. The PEI Questionnaire (PEI-Q) was developed through qualitative research with PEI patients and expert clinical input. This study evaluated the psychometric properties of the PEI-Q.

Improving women's quality of care for cardiovascular disease and diabetes: the feasibility and desirability of stratified reporting of objective performance measures.

Despite growing recognition of significant morbidity and mortality among women from cardiovascular disease, management of primary and secondary cardiac risk factors continues to be suboptimal for many women. Although there is a good deal of room to improve the care for cardiovascular disease and diabetes in men, existing gender differences in performance suggest much can be gained by specifically assessing and monitoring quality of care for these conditions in women. In this paper, we describe recent work showing gender differences in quality of ambulatory care in managed care plans with some plans having substantial gender differences on widely used measures of the quality of primary and secondary prevention of cardiac disease. We then discuss potential benefits of and barriers to routine reporting of objective measures of the quality of care, such as Health Plan Employer Data and Information Set (HEDIS) measures, by health plans.     

Antidepressant use and breast cancer risk

Summary Background Antidepressants are among the most commonly prescribed drugs in the United States. Laboratory studies suggest that because certain antidepressants increase prolactin levels that they may also increase breast cancer risk. However, human studies evaluating use of antidepressants in relation to breast cancer risk have yielded inconsistent results. Methods A population-based case-control study consisting of 975 breast cancer cases 65–79 years of age diagnosed from 1997–1999 and 1007 age and residence-matched controls was conducted in western Washington State. Detailed information on antidepressant use was obtained through structured in-person interviews. Logistic regression was performed to analyze the relationship between antidepressant use and breast cancer risk. Results Overall, there was no association between ever use of antidepressants and breast cancer risk (odds ratio [OR] = 1.2, 95% confidence interval [95% CI]: 0.9–1.6). When evaluated separately, tricyclic antidepressants (TCA), selective serotonin reuptake inhibitors (SSRI), and triazolopyridines were each not associated with breast cancer risk. However, risk varied by hormone receptor status. Compared to never users, ever users of SSRIs had elevated risks of progesterone receptor (PR) negative and estrogen receptor (ER) positive/PR-negative breast cancers (OR = 1.8, 95% CI: 1.1–3.6 and OR = 2.0, 95% CI: 1.1–3.8, respectively), but not of tumors with other hormone receptor profiles. Conclusions Based on these results and those of previous studies, there is limited evidence that any type of antidepressant use is associated with breast cancer risk overall. SSRIs may elevate risks of PR- and ER+/PR- tumors, though further studies are needed to confirm these associations.     

C-reactive protein predicts death in patients with non-ischemic cardiomyopathy

C-reactive protein (CRP) has been associated with atherosclerotic complications, and we hypothesized that CRP levels might also predict death in non-ischemic patients with left ventricular dysfunction. Two hundred and three patients with non-ischemic left ventricular dysfunction undergoing cardiac catheterization were included and were followed for 2.4 +/- 1.4 years to determine the incidence of fatal events. Death occurred in 15% of patients with low CRP (1st and 2nd tertiles) and 30% of patients with high CRP (3rd tertile). After adjustment for 11 covariates, high CRP (p = 0.037, hazard ratio = 2.0) significantly and independently predicted mortality. Even in the absence of coronary artery disease, patients with left ventricular dysfunction are at increased risk of mortality based on their baseline CRP concentrations.     

Oncogenes and Angiogenesis: down-regulation of thrombospondin-1 in normal fibroblasts exposed to factors from cancer cells harboring mutant ras

The onset of angiogenesis in cancer often involves down-regulation of endogenous angiogenesis inhibitors, of which thrombospondin-1 (TSP-1) is a paradigm. As this effect is thought to occur under the influence of transforming genetic lesions (e.g., expression of the mutant ras oncogene), its nature is regarded as intrinsic to cancer cells themselves. Here, we show that ras-transformed cancer cells can also induce TSP-1 down-regulation in their adjacent nontransformed stromal fibroblasts, but not in endothelial cells, in a paracrine and distance-dependent manner. Indeed, several H-ras-expressing fibrosarcoma (528ras1, B6ras, and NIH3T3Ras) and carcinoma (DLD-1 and IEC18Ras3) cells were found to release soluble factors capable of suppressing TSP-1 protein, mRNA, and promoter activity in nontumorigenic, immortalized dermal fibroblastic cell lines in culture (e.g., in fibroblasts expressing enhanced green fluorescent protein/TSP-1 reporter). This effect was abrogated in Id1-/- fibroblasts. At least two low molecular weight (<3 kDa), heat-labile, and trypsin-resistant mediators of TSP-1 suppression were found to be released from 528ras1 cells. Their effects on normal fibroblasts were inhibited (albeit to different extents) by pertussis toxin and, in one case, by dimethylsphingosine, none of which affected TSP-1 expression by 528ras1 cells. Collectively, our study suggests that the effect of mutant ras on tumor neovascularization is not limited to changes in angiogenic properties of cancer cells themselves. Rather, mutant ras, through a different signaling mechanism, may modulate the properties of the adjacent normal stroma, thus eliciting a proangiogenic field effect.     

A comparison of three methods of determination of energy density of elite figure skaters

Dietary energy density (kcal/g) is defined as available dietary energy per unit weight or volume of food. The consumption of energy-dense foods has been associated with increased obesity risk and with excessive weight gain. The objectives of this study were to compare how dietary energy density, calculated using three different methods relates to food choices and nutrient composition of the diets of elite figure skaters. Participants were 159 elite figure skaters attending training camps. Mean age was 18.4 y for boys (n = 79) and 15.9 y for girls (n = 80). Heights and weights were measured to calculate body-mass indices (BMI). Dietary intakes were based on 3-d food records analyzed using the Nutritionist IV program. Mean energy intakes were 2326 kcal/d for boys and 1545 kcal/d for girls. Dietary energy density,,based on foods and caloric beverages only, was 1.0 kcal/g. Dietary ED was positively associated with percent energy from fat and negatively with percent energy from sugar. The main sources of dietary energy in this group were baked goods, cereals, regular soda, low-fat milk, fruit juices, bagels and pizza. Percent energy from fast foods was associated with higher dietary energy density, whereas percent energy from dairy products, soft drinks, vegetables, and fruit was associated with lower dietary energy density. These results are consistent with past observations; higher energy density diets were higher in fat. In contrast, there was a negative relationship between sugar content and energy density of the diet.     

PEPFAR scale-up of pediatric HIV services: innovations, achievements, and challenges

HIV/AIDS has had a profound impact on children around the world since the start of the epidemic. There are currently 3.4 million children under the age of 15 years living with HIV globally, and more than 450,000 children currently receiving lifesaving antiretroviral treatment. This article describes efforts supported by the President's Emergency Plan for AIDS Relief (PEPFAR) to expand access to treatment for children living with HIV in high-burden countries. The article also highlights a series of case studies that illustrate the impact that the PEPFAR initiative has had on the pediatric HIV epidemic. Through its support of host governments and partner organizations, the PEPFAR initiative has expanded HIV testing and treatment for pregnant women to reduce vertical transmission of HIV, increased access to early infant diagnosis for HIV-exposed infants, improved training and resources for clinicians who provide pediatric care and antiretroviral treatment, and, through public-private partnerships with pharmaceutical manufacturers, helped increase the number of medications available for the treatment of HIV-infected children in resource-limited settings.     

Effect of antibiotic treatment on bacteriophage production by a cystic fibrosis epidemic strain of Pseudomonas aeruginosa

Phage production in response to antibiotics varied among four isolates of a Pseudomonas aeruginosa cystic fibrosis (CF) epidemic strain. Whereas ciprofloxacin induced higher levels of phage production, other CF-relevant antibiotics led to reduced production. We detected free phages directly in CF patient sputum samples by both plaque (40% positive) and PCR (76% positive) assays. Our observations suggest that the choice of antibiotics could influence the number of free phages within the CF lung environment.