A geospatiotemporal and causal inference epidemiological exploration of substance and cannabinoid exposure as drivers of rising US pediatric cancer rates

Background: Age-adjusted US total pediatric cancer incidence rates (TPCIR) rose 49% 1975–2015 for unknown reasons. Prenatal cannabis exposure has been linked with several pediatric cancers which together comprise the majority of pediatric cancer types. We investigated whether cannabis use was related spatiotemporally and causally to TPCIR. Methods: State-based age-adjusted TPCIR data was taken from the CDC Surveillance, Epidemiology and End Results cancer database 2003–2017. Drug exposure was taken from the nationally-representative National Survey of Drug Use and Health, response rate 74.1%. Drugs included were: tobacco, alcohol, cannabis, opioid analgesics and cocaine. This was supplemented by cannabinoid concentration data from the Drug Enforcement Agency and ethnicity and median household income data from US Census. Results: TPCIR rose while all drug use nationally fell, except for cannabis which rose. TPCIR in the highest cannabis use quintile was greater than in the lowest (β-estimate = 1.31 (95%C.I. 0.82, 1.80), P = 1.80 × 10− 7) and the time:highest two quintiles interaction was significant (β-estimate = 0.1395 (0.82, 1.80), P = 1.00 × 10− 14). In robust inverse probability weighted additive regression models cannabis was independently associated with TPCIR (β-estimate = 9.55 (3.95, 15.15), P = 0.0016). In interactive geospatiotemporal models including all drug, ethnic and income variables cannabis use was independently significant (β-estimate = 45.67 (18.77, 72.56), P = 0.0009). In geospatial models temporally lagged to 1,2,4 and 6 years interactive terms including cannabis were significant. Cannabis interactive terms at one and two degrees of spatial lagging were significant (from β-estimate = 3954.04 (1565.01, 6343.09), P = 0.0012). The interaction between the cannabinoids THC and cannabigerol was significant at zero, 2 and 6 years lag (from β-estimate = 46.22 (30.06, 62.38), P = 2.10 × 10− 8). Cannabis legalization was associated with higher TPCIR (β-estimate = 1.51 (0.68, 2.35), P = 0.0004) and cannabis-liberal regimes were associated with higher time:TPCIR interaction (β-estimate = 1.87 × 10− 4, (2.9 × 10− 5, 2.45 × 10− 4), P = 0.0208). 33/56 minimum e-Values were \u3e 5 and 6 were infinite. Conclusion: Data confirm a close relationship across space and lagged time between cannabis and TPCIR which was robust to adjustment, supported by inverse probability weighting procedures and accompanied by high e-Values making confounding unlikely and establishing the causal relationship. Cannabis-liberal jurisdictions were associated with higher rates of TPCIR and a faster rate of TPCIR increase. Data inform the broader general consideration of cannabinoid-induced genotoxicity

Re-Emergence of the Harmful Algal Bloom Species Alexandrium Monilatum in the Chesapeake Bay: Assessing Bloom Dynamics and Potential Health Impacts

Effective management of harmful algal blooms (HABs) within a region requires an understanding of species-specific HAB spatial and temporal distributions, bloom dynamics, as well as potential health impacts. In 2007, the southern Chesapeake Bay witnessed its first blooms of the HAB species Alexandrium monilatum. Since then, A. monilatum has bloomed in the region almost annually. A. monilatum produces the toxin ‘goniodomin A’ and is suspected in local mass mortalities of oyster larvae (Crassostrea virginica) grown for aquaculture and restoration projects. Representatives from Virginia’s multimillion dollar oyster aquaculture industry recently expressed great concern over A. monilatum impacts to their businesses; field and lab studies were designed to address these concerns. Sediment samples were collected from the southwest portion of the Chesapeake Bay in a systematic grid-sampling design to assess cyst (resting cell stage) distributions. Cysts were present in low densities at most sites, and cyst densities were high where blooms had been recorded in previous years. HAB toxicity bioassay methods developed at the Virginia Institute of Marine Science were modified to investigate adverse health impacts of five different A. monilatum cell density treatments on sub-adult oysters (~40-70 mm). Oysters delayed grazing when exposed to high densities (\u3e1000 cells/mL) of A. monilatum. Data from the 2015 bloom season and additional bioassays will be presented. Results from these studies could aid in the prediction of A. monilatum bloom severity and health effects on wild and aquacultured oysters, enabling development of best management practices to minimize impacts to the Virginia oyster industry

Classification and identification of Pfiesteria and Pfiesteria-like species

Dinoflagellates can be classified both botanically and zoologically; however, they are typically put in the botanical division Pyrrhophyta. As a group they appear most related to the protistan ciliates and apicomplexans at the ultrastructure level. Within the Pyrrhophyta are both unarmored and armored forms of the dominant, motile flagellated stage. Unarmored dinoflagellates do not have thecal or wall plates arranged in specific series, whereas armored species have plates that vary in thickness but are specific in number and arrangement. In armored dinoflagellates, the plate pattern and tabulation is a diagnostic character at the family, subfamily, and even genus levels. In most cases, the molecular characterization of dinoflagellates confirms the taxonomy on the basis of external morphology; this has been demonstrated for several groups. Together, both genetic and morphological criteria are becoming increasingly important for the characterization, separation, and identification of dinoflagellates species. Pfiesteria and Pfiesteria-like species are thinly armored forms with motile dinospore stages characterized by their distinct plate formulae. Pfiesteria piscicida is the best-known member of the genus; however, there is at least one other species. Other genetically and morphologically related genera, now grouped under the common names of Lucy, Shepherd\u27s crook, and cryptoperidiniopsoid, are being studied and described in separate works. All these other heterotrophic dinoflagellate groups, many of which are thought to be benign, co-occur in estuarine waters where Pfiesteria has been found

Evaluation of Crested Wheatgrass Introductions for Forage Yield and Quality

Crested wheatgrass, Agropyron cristatum and A. desertorum, are among the most important cool-season forage grasses in the United States and Canada, particularly for reseeding arid range sites. Further improvement in this grass by breeding depends on identifying sources of genetic variability for forage yield and quality. Foreign introductions are an obvious source of genetic variation since crested wheatgrasses are introduced species. In this study 38 accessions (PI lines) and 8 Nebraska experimental lines were evaluated for forage quality as measured by in vitro dry matter digestibility (IVDMD) and protein content and for forage yield. The cultivars \u27Ruff\u27 and \u27Nordan\u27 and two clonal lines were included as checks. The strains were evaluated at Lincoln and Alliance, NE., which differ markedly in climate. There were large differences among strains evaluated for all traits including first- and second-cut forage yield, IVDMD, protein content, heading date, height and first year basal spread. Utilization of PI 369167, PI 369170, Ruff, and Nordan in a breeding program could result in crested wheatgrasses with improved forage yield and quality

Monitoring young associations and open clusters with Kepler in two-wheel mode

We outline a proposal to use the Kepler spacecraft in two-wheel mode to monitor a handful of young associations and open clusters, for a few weeks each. Judging from the experience of similar projects using ground-based telescopes and the CoRoT spacecraft, this program would transform our understanding of early stellar evolution through the study of pulsations, rotation, activity, the detection and characterisation of eclipsing binaries, and the possible detection of transiting exoplanets. Importantly, Kepler's wide field-of-view would enable key spatially extended, nearby regions to be monitored in their entirety for the first time, and the proposed observations would exploit unique synergies with the GAIA ESO spectroscopic survey and, in the longer term, the GAIA mission itself. We also outline possible strategies for optimising the photometric performance of Kepler in two-wheel mode by modelling pixel sensitivity variations and other systematics.Comment: 10 pages, 6 figures, white paper submitted in response to NASA call for community input for alternative science investigations for the Kepler spacecraf

Out-Crossing Among Commercial Strains Of The Northern Quahog, Mercenaria Mercenaria: Survival, Growth And Implications For Selective Breeding

Because the accumulation of inbreeding within hatchery-propagated stocks of the hard clam, Mercenaria mercenaria (Linnaeus, 1758), could result in reduced growth and survival, we studied the potential for improving performance through out-crossing among existing hatchery strains. We produced all 10 possible out-crossed combinations among 5 strains of clams as well as all 5 pure parental strains simultaneously in the hatchery and measured their size at the time of metamorphosis (the spat stage) and at the end of a nursery period in mesh bags at a single field site (the seed stage). We then planted replicate plots of all fifteen strains at five field sites in Virginia, USA encompassing the range of salinity conditions used by commercial growers, and monitored growth and survival for two growing seasons. We found significant phenotypic differences among strains at the spat and seed stage, but different strains performed best at each stage. In the field we found significant site effects, strain effects and strain-by-site interactions, but there was no evidence of negative correlations in performance among sites indicating strong trade-offs that would be problematic for selective breeding. Three different linear contrasts designed to compare out-crossed and pure strains for each parental stock, test for nonadditive genetic effects within each pairing of different parental strains, and estimate the general combining ability of parental strains reveal a complex pattern. We found both inbreeding and out-breeding depression depending on the developmental stage of the clams and the parental strain examined. Within strain crosses generally produced larger spat but smaller seed. Out-crossed progeny were generally smaller at the spat stage than the average of their parental lines but larger at the seed stage. The two best performing parental strains had significant, positive, general combining abilities, whereas this measure was negative for the two worst parental strains. In the field, inbreeding depression was restricted to lines that showed poor pure strain performance, and these strains also showed poor general combining ability, whereas strains with good pure line performance showed out-breeding depression and good general combining ability. Only the poorest performing pure parental lines showed non-additive effects when we compared each out-crossed strain to the mean of its parental strains, suggesting that heterotic effects are unlikely to be useful for selective breeding. Finally, there were significant correlations between seed measurements and field performance indicating that it may be possible, in the context of selective breeding programs, to weed out inferior strains or families early in the life cycle

Skin ulcers in estuarine fishes: a comparative pathological evaluation of wild and laboratory-exposed fish.

The toxic dinoflagellate Pfiesteria piscicida Steidinger & Burkholder has recently been implicated as the etiologic agent of acute mass mortalities and skin ulcers in menhaden, Brevoortia tyrannus, and other fishes from mid-Atlantic U.S. estuaries. However, evidence for this association is largely circumstantial and controversial. We exposed tilapia (Oreochromis spp.) to Pfiesteria shumwayae Glasgow & Burkholder (identification based on scanning electron microscopy and molecular analyses) and compared the resulting pathology to the so-called Pfiesteria-specific lesions occurring in wild menhaden. The tilapia challenged by high concentrations (2,000-12,000 cells/mL) of P. shumwayaeexhibited loss of mucus coat and scales plus mild petecchial hemorrhage, but no deeply penetrating chronic ulcers like those in wild menhaden. Histologically, fish exhibited epidermal erosion with bacterial colonization but minimal associated inflammation. In moribund fish, loss of epidermis was widespread over large portions of the body. Similar erosion occurred in the mucosa lining the oral and branchial cavities. Gills exhibited epithelial lifting, loss of secondary lamellar structure, and infiltration by lymphoid cells. Epithelial lining of the lateral line canal (LLC) and olfactory organs exhibited severe necrosis. Visceral organs, kidney, and neural tissues (brain, spinal cord, ganglia, peripheral nerves) were histologically normal. An unexpected finding was the numerous P. shumwayae cells adhering to damaged skin, skin folds, scale pockets, LLC, and olfactory tissues. In contrast, histologic evaluation of skin ulcers in over 200 wild menhaden from Virginia and Maryland portions of the Chesapeake Bay and the Pamlico Estuary, North Carolina, revealed that all ulcers harbored a deeply invasive, highly pathogenic fungus now known to be Aphanomyces invadans. In menhaden the infection always elicited severe myonecrosis and intense granulomatous myositis. The consistent occurrence of this fungus and the nature and severity of the resulting inflammatory response indicate that these ulcers are chronic (age >1 week) and of an infectious etiology, not the direct result of an acute toxicosis initiated by Pfiesteria toxin(s) as recently hypothesized. The disease therefore is best called ulcerative mycosis (UM). This study indicates that the pathology of Pfiesteria laboratory exposure is fundamentally different from that of UM in menhaden; however, we cannot rule out Pfiesteria as one of many possible early initiators predisposing wild fishes to fungal infection in some circumstances

Microsatellite marker development and analysis in the eastern oyster (Crassostrea virginica): Confirmation of null alleles and non-Mendelian segregation ratios

Eighteen microsatellite markers were developed for the Crassostrea virginica nuclear genome, including di-, tri-, and tetranucleotide microsatellite repeat regions that included perfect, imperfect, and compound repeat sequences. A reference panel with DNA from the parents and four progeny of 10 full-sib families was used for a preliminary confirmation of polymorphism at these loci and indications of null alleles. Null alleles were discovered at three loci; in two instances, primer redesign enabled their amplification. Two to five representative alleles from each locus were sequenced to ensure that the targeted loci were amplifying. The sequence analysis revealed not only variation in the number of simple sequence repeat units, but also polymorphisms in the microsatellite flanking regions. A total of 3626 bp of combined microsatellite flanking region from the 18 loci was examined, revealing indels as well as nucleotide site substitutions. Overall, 16 indels and 146 substitutions were found with an average of 4.5% polymorphism across all loci. Eight markers were tested on the parents and 39-61 progeny from each of four families for examination of allelic inheritance patterns and genotypic ratios. Twenty-six tests of segregation ratios revealed eight significant departures from expected Mendelian ratios, three of which remained significant after correction for multiple tests. Deviations were observed in both the directions of heterozygote excess and deficiency

Mycobacterium pseudoshottsii sp nov., a slowly growing chromogenic species isolated from Chesapeake Bay striped bass (Morone saxatilis)

A group of slowly growing photochromogenic mycobacteria was isolated from Chesapeake Bay striped bass (Morone saxatilis) during an epizootic of mycobacteriosis. Growth characteristics, acid-fastness and 16S rRNA gene sequencing results were consistent with those of the genus Mycobacterium, Biochemical reactions, growth characteristics and mycolic acid profiles (HPLC) resembled those of Mycobacterium shottsii, a non-pigmented mycobacterium also isolated during the same epizootic. Sequencing of the 16S rRNA genes, the gene encoding the exported repeated protein (erp) and the gene encoding the 65 kDa heat-shock protein (hsp65) and restriction enzyme analysis of the hsp65 gene demonstrated that this group of isolates is unique. Insertion sequences associated with Mycobacterium ulcerans, IS2404 and IS2606, were detected by PCR. These isolates could be differentiated from other slowly growing pigmented mycobacteria by their inability to grow at 37 degrees C, production of niacin and urease, absence of nitrate reductase, negative Tween 80 hydrolysis and resistance to isoniazid (1 mu g ml(-1)), p-nitrobenzoic acid, thiacetazone and thiophene-2-carboxylic hydrazide. On the basis of this polyphasic study, it is proposed that these isolates represent a novel species, Mycobacterium pseudoshottsii sp. nov. The type strain, L15(T), has been deposited in the American Type Culture Collection as ATCC BAA-883(T) and the National Collection of Type Cultures (UK) as NCTC 13318(T)

Parasitic Dinoflagellate \u3ci\u3eHematodinium perezi\u3c/i\u3e Prevalence in Larval and Juvenile Blue Crabs \u3ci\u3eCallinectes sapidus\u3c/i\u3e from Coastal Bays of Virginia

The parasitic dinoflagellate Hematodinium perezi infects the American blue crab Callinectes sapidus and other decapods along the Eastern seaboard and Gulf of Mexico coast of the USA. Large juvenile and adult blue crabs experience high mortality during seasonal outbreaks of H. perezi, but less is known about its presence in the early life history stages of this host. We determined the prevalence of H. perezi in megalopae and early benthic juvenile crabs from multiple locations along the Virginia portion of the Delmarva Peninsula. The DNA of H. perezi was not detected in any megalopae collected from several locations within the oceanic coastal bay complex in which H. perezi is found at high prevalence levels. However, prevalence levels were high in early benthic juveniles from 2 oceanic coastal embayments: South Bay and Cobb Bay. Prevalence levels were lower at locations within Chesapeake Bay, including Cherrystone Creek, Hungars Creek, and Pungoteague Creek. Sampling over different seasons and several consecutive years indicates that disease transmission occurs rapidly after megalopae settle in high-salinity bays along the Delmarva Peninsula during the late summer and fall. Infected juvenile crabs can overwinter with the parasite and, when subjected to increasing water temperatures in spring, infections progress rapidly, culminating in transmission to other crabs in late spring and early summer. In high-salinity embayments, H. perezi can reach high prevalence levels and may significantly affect recruitment of juvenile blue crabs into the adult fishery