Organically modified silica nanoparticles are biocompatible and can be targeted to neurons in vivo

The application of nanotechnology in biological research is beginning to have a major impact leading to the development of new types of tools for human health. One focus of nanobiotechnology is the development of nanoparticle-based formulations for use in drug or gene delivery systems. However most of the nano probes currently in use have varying levels of toxicity in cells or whole organisms and therefore are not suitable for in vivo application or long-term use. Here we test the potential of a novel silica based nanoparticle (organically modified silica, ORMOSIL) in living neurons within a whole organism. We show that feeding ORMOSIL nanoparticles to Drosophila has no effect on viability. ORMOSIL nanoparticles penetrate into living brains, neuronal cell bodies and axonal projections. In the neuronal cell body, nanoparticles are present in the cytoplasm, but not in the nucleus. Strikingly, incorporation of ORMOSIL nanoparticles into the brain did not induce aberrant neuronal death or interfered with normal neuronal processes. Our results in Drosophila indicate that these novel silica based nanoparticles are biocompatible and not toxic to whole organisms, and has potential for the development of long-term applications.     

Mechanisms involved in the intestinal absorption of dietary vitamin A and provitamin A carotenoids

Vitamin A is an essential nutrient for humans and is converted to the visual chromophore, 11-cis-retinal, and to the hormone, retinoic acid. Vitamin A in animal-derived foods is found as long chain acyl esters of retinol and these are digested to free fatty acids and retinol before uptake by the intestinal mucosal cell. The retinol is then reesterified to retinyl esters for incorporation into chlylomicrons and absorbed via the lymphatics or effluxed into the portal circulation facilitated by the lipid transporter, ABCA1. Provitamin A carotenoids such as β-carotene are found in plant-derived foods. These and other carotenoids are transported into the mucosal cell by scavenger receptor class B type I (SR-BI). Provitamin A carotenoids are partly converted to retinol by oxygenase and reductase enzymes and the retinol so produced is available for absorption via the two pathways described above. The efficiency of vitamin A and carotenoid intestinal absorption is determined by the regulation of a number of proteins involved in the process. Polymorphisms in genes for these proteins lead to individual variability in the metabolism and transport of vitamin A and carotenoids. This article is part of a Special Issue entitled Retinoid and Lipid Metabolism.     

Xanthophylls are preferentially taken up compared with beta-carotene by retinal cells via a SRBI-dependent mechanism

The purpose of this study was to investigate the mechanisms by which carotenoids [xanthophylls vs. beta-carotene(beta-C)] are taken up by retinal pigment epithelial (RPE) cells. The human RPE cell line, ARPE-19, was used. When ARPE-19 cells were fully differentiated (7-9 weeks), the xanthophylls lutein (LUT) and zeaxanthin (ZEA) were taken up by cells to an extent 2-fold higher than beta-C (P < 0.05). At 9 weeks, cellular uptakes were 1.6, 2.5, and 3.2%, respectively, for beta-C, LUT, and ZEA. Similar extents were observed when carotenoids were delivered in either Tween 40 or "chylomicrons" produced by Caco-2 cells. Differentiated ARPE-19 cells did not exhibit any detectable beta-C 15,15'-oxygenase activity or convert exogenous beta-C into vitamin A. When using specific antibodies against the lipid transporters cluster determinant 36 (CD36) and scavenger receptor class B type I (SR-BI), cellular uptake of beta-C and ZEA were significantly decreased (40-60%) with anti-SR-BI but not with anti-CD36. Small interfering RNA transfection for SR-BI led to marked knockdown of SR-BI protein expression (approximately 90%), which resulted in decreased beta-C and ZEA uptakes by 51% and 87%, respectively. Thus, the present data show that RPE cells preferentially take up xanthophylls versus the carotene by a process that appears to be entirely SR-BI-dependent for ZEA and partly so for beta-C. This mechanism may explain, in part, the preferential accumulation of xanthophylls in the macula of the retina.     

Antibody profiling by proteome microarray reveals the immunogenicity of the attenuated smallpox vaccine modified vaccinia virus ankara is comparable to that of Dryvax

Modified vaccinia virus Ankara (MVA) is a highly attenuated vaccinia virus that is under consideration as an alternative to the conventional smallpox vaccine Dryvax. MVA was attenuated by extensive passage of vaccinia virus Ankara in chicken embryo fibroblasts. Several immunomodulatory genes and genes that influence host range are deleted or mutated, and replication is aborted in the late stage of infection in most nonavian cells. The effect of these mutations on immunogenicity is not well understood. Since the structural genes appear to be intact in MVA, it is hypothesized that critical targets for antibody neutralization have been retained. To test this, we probed microarrays of the Western Reserve (WR) proteome with sera from humans and macaques after MVA and Dryvax vaccination. As most protein sequences of MVA are 97 to 99% identical to those of other vaccinia virus strains, extensive binding cross-reactivity is expected, except for those deleted or truncated. Despite different hosts and immunization regimens, the MVA and Dryvax antibody profiles were broadly similar, with antibodies against membrane and core proteins being the best conserved. The responses to nonstructural proteins were less well conserved, although these are not expected to influence virus neutralization. The broadest antibody response was obtained for hyperimmune rabbits with WR, which is pathogenic in rabbits. These data indicate that, despite the mutations and deletions in MVA, its overall immunogenicity is broadly comparable to that of Dryvax, particularly at the level of antibodies to membrane proteins. The work supports other information suggesting that MVA may be a useful alternative to Dryvax.     

Obesity and Self‐reported General Health,Hawaii BRFSS: Are Polynesians at Higher Risk?

Objective: This study compared the relationship between fair/poor general health status among overweight and obese Polynesians with that among other overweight and obese persons in Hawaii. Methods and Procedures: Data were pooled from the 1998–2003 Hawaii Behavioral Risk Factor Surveillance System (BRFSS) and logistic regression used to examine the predictors of fair/poor health status. Results: Polynesians were significantly more likely to be obese than non‐Polynesians; overweight Polynesians were more likely than other overweight individuals to report fair/poor health status. After adjusting for confounders, among Polynesians, being obese was no longer associated with fair/poor health. Non‐Polynesians who were obese (odds ratio 1.9; 95% confidence interval: 1.4–2.6), older, less educated, smokers, diabetic, hypertensive, and physically inactive were more likely to report fair/poor health. Discussion: Although Polynesians were significantly more obese than the rest of the Hawaii population, their weight was not independently associated with their odds for fair/poor health as it was with non‐Polynesians. The difference may be that, for Polynesians, hypertension and diabetes overrode the effect of obesity on general health status or this group maintains different cultural perceptions of body size. Regardless, these findings show a major health risk among Polynesians and suggest the need for culturally specific health interventions.     

Self‐Reported Body Mass Index and Health‐Related Quality of Life: Findings from the Behavioral Risk Factor Surveillance System

Objective: To examine the relationship between self‐reported body mass index (BMI) and health‐related quality of life in the general adult population in the United States. Research Methods and Procedures: Using data from 109,076 respondents in the 1996 Behavioral Risk Factor Surveillance System, we examined how self‐reported BMI is associated with five health‐related quality of life measures developed by the Centers for Disease Control and Prevention for population health surveillance. Results: After adjusting for age, gender, race or ethnicity, educational attainment, employment status, smoking status, and physical activity status, participants with a self‐reported BMI of <18.5 kg/m² and participants with a self‐reported BMI of ≥30 kg/m² reported impaired quality of life. Compared with persons with a self‐reported BMI of 18.5 to <25 kg/m², odds ratios (ORs) of poor or fair self‐rated health increased among persons with self‐reported BMIs of <18.5 (1.57, 95% confidence interval [CI]: 1.31 to 1.89), 25 to <30 kg/m² (1.12, 95% CI: 1.04 to 1.20), 30 to <35 kg/m² (1.65, 95% CI: 1.50 to 1.81), 35 to <40 kg/m² (2.58, 95% CI: 2.21 to 3.00), and ≥40 kg/m² (3.23, 95% CI: 2.63 to 3.95); ORs for reporting ≥14 days of poor physical health during the previous 30 days were 1.44 (95% CI: 1.21 to 1.72), 1.04 (95% CI: 0.96 to 1.14), 1.32 (95% CI: 1.19 to 1.47), 1.80 (95% CI: 1.52 to 2.13), and 2.37 (95% CI: 1.90 to 2.94), respectively; ORs for having ≥14 days of poor mental health during the previous 30 days were 1.18 (95% CI: 0.97 to 1.42), 1.02 (95% CI: 0.95 to 1.11), 1.22 (95% CI: 1.10 to 1.36), 1.68 (95% CI: 1.42 to 1.98), and 1.66 (95% CI: 1.32 to 2.09), respectively. Discussion: In the largest study to date, low and increased self‐reported BMI significantly impaired health‐related quality of life. Particularly, deviations from normal BMI affected physical functioning more strongly than mental functioning.     

Genetic Evidence that the uvsE Gene Product of Deinococcus radiodurans R1 Is a UV Damage Endonuclease

An in vitro transposition system, developed to facilitate gene disruption in Deinococcus radiodurans R1, has been used to inactivate the gene designated dr1819 in uvrA-1(+) and uvrA-1 backgrounds. dr1819 encodes a protein with homology to a UV DNA damage endonuclease expressed by Schizosaccharomyces pombe. Interruption of dr1819 greatly sensitizes the uvrA-1 strain but not the uvrA-1(+) strain to UV light, indicating that the dr1819 gene product is a component in a DNA repair pathway that can compensate for the loss of nucleotide excision repair in this species. Clones of dr1819 will restore UV resistance to UVS78, a uvrA-1 uvsE strain, indicating that dr1819 and uvsE are the same locus.     

Assessment of skin carbonyl content as a noninvasive measure of biological age.

The oxidative modification of proteins by reactive species, especially reactive oxygen species, is implicated in the etiology or progression of a panoply of disorders and diseases. For the most part, oxidatively modified proteins are not repaired and must be removed by proteolytic degradation. The level of these modified molecules can be quantitated by measurement of the protein carbonyl content, which has been shown to increase in a variety of diseases and processes, most notably during aging. However, these studies have required invasive techniques to obtain cells for analysis. We examined the possibility that desquamating skin cells (corneocytes) would also show an age-related increase in protein carbonyl content, thus providing a noninvasive method for assessing biological age. This was not the case, as we found no age-dependent relationship in the protein carbonyl content of skin cells from volunteers aged 20 to 79 years.     

Parasitism of Leptasterias spp. (Echinodermata: Asteroidea) by the ciliated protozoan Orchitophrya stellarum (Scuticociliata)

Abstract. The parasitic ciliate Orchitophrya stellarum was found in the testes of brooding, winter-spawning Leptasterias spp. from San Juan Island, Washington, but not in the testes of Leptasterias spp. from the Lynn Canal, Alaska. Dense populations of the ciliate were localized within the fuller areas of testes, where sperm counts were significantly reduced. The ciliates were loaded with phagosomes, some of which contained sperms in various stages of digestion. Leptasterias spp. are not as severely impacted by this ciliated protozoan parasite as Pisaster ochraceus. Leptasterias spp. may serve as seasonal hosts for O. stellarum ; this ciliate lives in seawater and in ripe males of winter-brooding and spring-summer broadcasting sea stars. The wet weight of parasitized and non-parasitized males did not differ. The sex ratio did not deviate from the expected (1:1) in any population except at Point Louisa, Alaska, where males outnumbered females. Testis indexes of parasitized males from 2 of the 4 locations on San Juan Island, Washington were significantly reduced relative to non-parasitized males, indicating a loss of sperm output during spawning.