Measurement of spatial and temporal variation in volatile hazardous air pollutants in Tacoma, Washington, using a mobile membrane introduction mass spectrometry (MIMS) system.

The objective of this study was to use membrane introduction mass spectrometry (MIMS), implemented on a mobile platform, in order to provide real-time, fine-scale, temporally and spatially resolved measurements of several hazardous air pollutants. This work is important because there is now substantial evidence that fine-scale spatial and temporal variations of air pollutant concentrations are important determinants of exposure to air pollution and adverse health outcomes. The study took place in Tacoma, WA during periods of impaired air quality in the winter and summer of 2008 and 2009. Levels of fine particles were higher in winter compared to summer, and were spatially uniform across the study area. Concentrations of vapor phase pollutants measured by membrane introduction mass spectrometry (MIMS), notably benzene and toluene, had relatively uniform spatial distributions at night, but exhibited substantial spatial variation during the day-daytime levels were up to 3-fold higher at traffic-impacted locations compared to a reference site. Although no direct side-by-side comparison was made between the MIMS system and traditional fixed site monitors, the MIMS system typically reported higher concentrations of specific VOCs, particularly benzene, ethylbenzene and naphthalene, compared to annual average concentrations obtained from SUMA canisters and gas chromatographic analysis at the fixed sites.

Biomonitoring of estrogen and melatonin metabolites among women residing near radio and television broadcasting transmitters.

OBJECTIVES: Metabolites of estrogen (estrone-3-glucuronide [E1G]) and melatonin (6-hydroxymelatonin sulfate [6-OHMS]) were characterized among women living in a community with increased radiofrequency (RF) exposure from radio and television transmitters. METHODS: RF spot measurements, and personal 60-Hz magnetic field and residential parameters were collected. Overnight urine samples were assayed for E1G and 6-OHMS excretion. RESULTS: Among premenopausal women, there were no associations between RF or 60-Hz nonionizing radiation and E1G or 6-OHMS excretion. Among postmenopausal women, increased residential RF exposures, transmitter proximity and visibility, and temporally stable 60-Hz exposures were significantly associated with increased E1G excretion. This association was strongest among postmenopausal women with low overnight 6-OHMS levels. CONCLUSIONS: RF and temporally stable 60-Hz exposures were associated with increased E1G excretion among postmenopausal women. Women with reduced nocturnal 6-OHMS excretion may represent a sensitive subgroup.

Comparison of global positioning system (GPS) tracking and parent-report diaries to characterize children's time-location patterns.

Respondent error, low resolution, and study participant burden are known limitations of diary timelines used in exposure studies such as the National Human Exposure Assessment Survey (NHEXAS). Recent advances in global positioning system (GPS) technology have produced tracking devices sufficiently portable, functional and affordable to utilize in exposure assessment science. In this study, a differentially corrected GPS (dGPS) tracking device was compared to the NHEXAS diary timeline. The study also explored how GPS can be used to evaluate and improve such diary timelines by determining which location categories and which respondents are least likely to record "correct" time-location responses. A total of 31 children ages 3-5 years old wore a dGPS device for all waking hours on a weekend day while their parents completed the NHEXAS diary timeline to document the child's time-location pattern. Parents misclassified child time-location approximately 48% of the time using the NHEXAS timeline in comparison to dGPS. Overall concordance between methods was marginal (kappa=0.33-0.35). The dGPS device found that on average, children spent 76% of the 24-h study period in the home. The diary underestimated time the child spent in the home by 17%, while overestimating time spent inside other locations, outside at home, outside in other locations, and time spent in transit. Diary data for time spent outside at home and time in transit had the lowest response concordance with dGPS. The diaries of stay-at-home mothers and mothers working unskilled labor jobs had lower concordance with dGPS than did those of the other participants. The ability of dGPS tracking to collect continuous rather than categorical (ordinal) data was also demonstrated. It is concluded that automated GPS tracking measurements can improve the quality and collection efficiency of time-location data in exposure assessment studies, albeit for small cohorts.

Radio frequency nonionizing radiation in a community exposed to radio and television broadcasting.

Exposure to radio frequency (RF) nonionizing radiation from telecommunications is pervasive in modern society. Elevated disease risks have been observed in some populations exposed to radio and television transmissions, although findings are inconsistent. This study quantified RF exposures among 280 residents living near the broadcasting transmitters for Denver, Colorado. RF power densities outside and inside each residence were obtained, and a global positioning system (GPS) identified geographic coordinates and elevations. A view-shed model within a geographic information system (GIS) characterized the average distance and percentage of transmitters visible from each residence. Data were collected at the beginning and end of a 2.5-day period, and some measurements were repeated 8-29 months later. RF levels logged at 1-min intervals for 2.5 days varied considerably among some homes and were quite similar among others. The greatest differences appeared among homes within 1 km of the transmitters. Overall, there were no differences in mean residential RF levels compared over 2.5 days. However, after a 1- to 2-year follow-up, only 25% of exterior and 38% of interior RF measurements were unchanged. Increasing proximity, elevation, and line-of-sight visibility were each associated with elevated RF exposures. At average distances from > 1-3 km, exterior RF measurements were 13-30 times greater among homes that had > 50% of the transmitters visible compared with homes with < or = 50% visibility at those distances. This study demonstrated that both spatial and temporal factors contribute to residential RF exposure and that GPS/GIS technologies can improve RF exposure assessment and reduce exposure misclassification.

Detection of an RNA-dependent RNA polymerase in mitochondria from a mitovirus-infected isolate of the Dutch Elm disease fungus, Ophiostoma novo-ulmi.

RNA-dependent RNA polymerase (RdRp) activity has been detected in mitochondria from an isolate of Ophiostoma novo-ulmi infected with O. novo-ulmi mitovirus 6 (OnuMV6). The reaction products corresponded to the double-stranded and single-stranded forms of OnuMV6 RNA. Western blot analysis using antibodies raised against a conserved RdRp region has detected a protein of ca. 80 kDa in OnuMV6-infected mitochondria, close to the predicted size of the OnuMV6 RdRp. No RdRp activity or protein was detected in mitochondria from an uninfected O. novo-ulmi isolate. This is the first detection of a virus RdRp in fungal mitochondria and the results are consistent with the use of UGA tryptophan codons in its synthesis.

Multiple mitochondrial viruses in an isolate of the Dutch Elm disease fungus Ophiostoma novo-ulmi.

The nucleotide sequences of three mitochondrial virus double-stranded (ds) RNAs, RNA-4 (2599 nucleotides), RNA-5 (2474 nucleotides), and RNA-6 (2343 nucleotides), in a diseased isolate Log1/3-8d2 (Ld) of the Dutch elm disease fungus Ophiostoma novo-ulmi have been determined. All these RNAs are A-U-rich (71-73% A + U residues). Using the fungal mitochondrial genetic code in which UGA codes for tryptophan, the positive-strand of each of RNAs 4, 5, and 6 contains a single open reading frame (ORF) with the potential to encode a protein of 783, 729, and 695 amino acids, respectively, all of which contain conserved motifs characteristic of RNA-dependent RNA polymerases (RdRps). Sequence comparisons showed that these RNAs are related to each other and to a previously characterized RNA, RNA-3a, from the same O. novo-ulmi isolate, especially within the RdRp-like motifs. However, the overall RNA nucleotide and RdRp amino acid sequence identities were relatively low (43-55% and 20-32%, respectively). The 5'- and 3'-terminal sequences of these RNAs are different, but they can all be folded into potentially stable stem-loop structures. Those of RNA-4 and RNA-6 have inverted complementarity, potentially forming panhandle structures. Their molecular and biological properties indicate that RNAs 3a, 4, 5, and 6 are the genomes of four different viruses, which replicate independently in the same cell. These four viruses are also related to a mitochondrial RNA virus from another fungus, Cryphonectria parasitica, recently designated the type species of the Mitovirus genus of the Narnaviridae family, and to a virus from the fungus Rhizoctonia solani. It is proposed that the four O. novo-ulmi mitochondrial viruses are assigned to the Mitovirus genus and designated O. novo-ulmi mitovirus (OnuMV) 3a-Ld, 4-Ld, 5-Ld, and 6-Ld, respectively. Northern blot analysis indicated that O. novo-ulmi Ld nucleic acid extracts contain more single-stranded (ss, positive-stranded) RNA than dsRNA for all three newly described mitoviruses. O. novo-ulmi RNA-7, previously believed to be a satellite-like RNA, is shown to be a defective RNA, derived from OnuMV4-Ld RNA by multiple internal deletions. OnuMV4-Ld is therefore the helper virus for the replication of both RNA-7 and another defective RNA, RNA-10. Sequence comparisons indicate that RNA-10 could be derived from RNA-7, as previously suggested, or derived directly from RNA-4.

Evolutionary relationships among putative RNA-dependent RNA polymerases encoded by a mitochondrial virus-like RNA in the Dutch elm disease fungus, Ophiostoma novo-ulmi, by other viruses and virus-like RNAs and by the Arabidopsis mitochondrial genome.

The nucleotide sequence (2617 nucleotides) of virus-like double-stranded (ds) RNA 3a in a diseased isolate, Log1/3-8d2 (Ld), of the ascomycete fungus Ophiostoma novo-ulmi has been determined. One strand of the dsRNA contains an open reading frame (ORF) with the potential to encode a protein of 718 amino acids, and the complementary strand contains two smaller ORFs with the potential to encode proteins of 178 and 182 amino acids, respectively. The large ORF contains 12 UGA codons which code for tryptophan in ascomycete mitochondria and has a codon bias typical of mitochondrial genes, consistent with the localization of Ld dsRNAs within the mitochondria. The amino acid sequence contains motifs characteristic of RNA-dependent RNA polymerases (RdRps). This putative RdRp was shown to be related to putative RdRps of mitochondrial dsRNAs of another ascomycete and a basidiomycete fungus and also to a putative RdRp encoded by the mitochondrial genome of Arabidopsis thaliana. In multiple sequence alignments, the fungal mitochondrial dsRNA-encoded RdRp-like proteins formed a cluster, ancestrally related to the RdRps of the yeast 20S and 23S RNA replicons and of the positive-stranded RNA bacteriophages of the Leviviridae family, but distinct from RdRps of other families and genera of fungal RNA viruses and related plant and animal RNA viruses. Northern blot analysis with RNA 3a strand-specific probes indicated that nucleic acid extracts of Ld contain more single-stranded (positive-stranded) RNA than dsRNA, consistent with an evolutionary relationship between RNA 3a and positive-stranded RNA phages.

Novel structures of two virus-like RNA elements from a diseased isolate of the Dutch elm disease fungus, Ophiostoma novo-ulmi.

The nucleotide sequences of 2 of the 10 mitochondrial double-stranded (ds) RNA segments in a diseased isolate, Log 1/3-8d2 (Ld), of Ophiostoma novo-ulmi, RNA-7 (1057 nucleotides) and RNA-10 (317-330 nucleotides), have been determined. Both RNAs are A-U-rich, but in Southern and Northern blots, no hybridization with mitochondrial DNA or RNA could be detected. Only very short open reading frames were found in both RNAs. As most of its sequence is unrelated to any of the other Ld dsRNAs, RNA-7 may be regarded as a satellite RNA. Northern blotting detected a full-length single-stranded (ss) form of RNA-7 in nucleic acid extracts from Ld. The 5'- and 3'-terminal 39 nucleotides of ssRNA-7 are imperfect inverted complementary repeats of each other, which could cause ssRNA-7 to form a panhandle structure. In addition, the 5'-terminal nucleotides 1-28 and 3'-terminal nucleotides 1032-1057 of ssRNA-7 each contained inverted complementary sequences, allowing the possibility for each terminus to form separate stem-loop structures. The combination of these two structural features has not been found previously in any dsRNA or ssRNA virus. RNA-10 was shown to have an unusual structure, consisting of a mosaic of sequences derived from regions of the 5'- and 3'-termini, or just the 5'-terminus, of RNA-7, RNA-10 has a high degree of inverted complementarity, with the potential to be folded into a very stable hairpin structure. A model for the formation of RNA-10 is presented, involving replicase-driven strand switching between (-)-strand and (+)-strand templates during RNA synthesis, followed by utilization of the nascent strand as a primer and template to form a snap-back RNA.

Flavones. 2. Synthesis and structure-activity relationship of flavodilol and its analogues, a novel class of antihypertensive agents with catecholamine depleting properties.

(3-Phenyl-7-flavonoxy)propanolamines have been shown to exhibit antihypertensive activity in spontaneously hypertensive rats. Although they are structurally similar to classical beta-adrenergic blocking compounds, their activity is not due to inhibition of beta-adrenoceptors. In the present study, a series of simple flavonoxypropanolamines was prepared to further explore the structural requirements for the antihypertensive effect of these compounds. A structure-activity relationship of these derivatives indicates that the position of the oxypropanolamine side chain, the hydroxy group of the side chain, steric bulkiness and length of N substituents, degree of the N-substitution, phenyl group at the 2-position of the chromone nucleus, and substituents of the phenyl group or B ring of the flavone play significant roles in imparting pharmacological effects. In addition, there is a good correlation between the antihypertensive activity and depletion of myocardial norepinephrine. Of these analogues tested, the most effective one was flavodilol. Only the 8-substituted analogue 6 was found to be a beta-antagonist. Flavodilol was chosen for in-depth pharmacological, toxicological, and clinical evaluation.

Flavones. 1. Synthesis and antihypertensive activity of (3-phenylflavonoxy)propanolamines without beta-adrenoceptor antagonism.

The synthesis of a series of (3-phenylflavonoxy)propanolamines is described. These compounds were evaluated for potential antihypertensive activity in spontaneously hypertensive rats, as well as for in vivo and in vitro evidence of beta-adrenoceptor antagonism. Some of the compounds of this series exhibited effective antihypertensive properties but did not antagonize beta-adrenergic receptors. These active compounds represent a unique series of effective antihypertensive agents that, despite possessing structural characteristics typical of beta-blockers, does not have beta-adrenergic receptor blocking activity.