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Humpback whales (Megaptera novaeangliae) exhibit maternally driven fidelity to feeding grounds, and yet occasionally occupy new areas. Humpback whale sightings and mortalities in the New York Bight apex (NYBA) have been increasing over the last decade, providing an opportunity to study this phenomenon in an urban habitat. Whales in this area overlap with human activities, including busy shipping traffic leading into the Port of New York and New Jersey. The site fidelity, population composition and demographics of individual whales were analysed to better inform management in this high-risk area. Whale watching and other opportunistic data collections were used to identify 101 individual humpback whales in the NYBA from spring through autumn, 2012–2018. Although mean occurrence was low (2.5 days), mean occupancy was 37.6 days, and 31.3% of whales returned from one year to the next. Individuals compared with other regional and ocean-basin-wide photo-identification catalogues (N = 52) were primarily resighted at other sites along the US East Coast, including the Gulf of Maine feeding ground. Sightings of mother-calf pairs were rare in the NYBA, suggesting that maternally directed fidelity may not be responsible for the presence of young whales in this area. Other factors including shifts in prey species distribution or changes in population structure more broadly should be investigated.

Wild mushrooms, while largely explored for their ecological significance, have not been systematically studied for their medicinal properties. This is the first report of biological activities of mushrooms from Haida Gwaii, British Columbia, Canada. The 17 mushroom species in this study were collected from multiple locations on Haida Gwaii and were screened for antiproliferative, immunostimulatory, and anti-inflammatory activities. Prior to screening, mushrooms were genetically identified and then sequentially fractionated into four crude extracts using 80% ethanol, 50% methanol, water, and 5% sodium hydroxide. We report here the strong antiproliferative and antiinflammatory activities of Amanita augusta, Phellodon atratus, Guepinia helvelloides, Chroogomphus tomentosus, Laetiporus conifericola, and Inocybe sp. In addition, A. augusta, G. helvelloides, and Inocybe sp. showed potent immunostimulatory activity. Two other species (Ganoderma tsugae and Pleurotus ostreatus) displayed strong immunostimulatory activity consistent with previous reports by others, suggesting that similar constituents are present in the same species from Haida Gwaii. For nine species (Russula paludosa, Hygrophoropsis aurantiaca, Tricholomopsis rutilans, Tyromyces chioneus, Hydnum repandum, Hypholoma fasciculare, Clavulina cinerea, P. ostreatus and Ramaria cystidiophora), we describe antiproliferative, immunostimulatory, and/or anti-inflammatory activities that have never been reported before. The new findings serve as a platform for future investigations into the potentially novel bioactive constituents of these mushrooms, as well as an incentive to further study a wider array of wild mushrooms for medicinal properties.

In our search for bioactive mushrooms native to British Columbia, we determined that the ethanol extracts from fruiting bodies of the terrestrial polypore Albatrellus flettii had potent anti-cell viability activity. Using bioassay-guided fractionation, mass spectrometry and nuclear magnetic resonance, we successfully isolated three known compounds (grifolin, neogrifolin and confluentin). These compounds represent the major anti-cell viability components from the ethanol extracts of A. flettii. We also identified a novel biological activity for these compounds, specifically in down-regulating KRAS expression in two human colon cancer cell lines. Relatively little is known about the anti-cell viability activity and mechanism of action of confluentin. For the first time, we show the ability of confluentin to induce apoptosis and arrest the cell cycle at the G2/M phase in SW480 human colon cancer cells. The oncogenic insulin-like growth factor 2 mRNA-binding protein 1 (IMP1) has been previously shown to regulate KRAS mRNA expression in colon cancer cells, possibly through its ability to bind to the KRAS transcript. Using a fluorescence polarization assay, we show that confluentin dose-dependently inhibits the physical interaction between KRAS RNA and full-length IMP1. The inhibition also occurs with truncated IMP1 containing the KH1 to KH4 domain (KH1to4 IMP1), but not with the di-domain KH3 and KH4 (KH3&4 IMP1). In addition, unlike the control antibiotic neomycin, grifolin, neogrifolin and confluentin do not bind to KRAS RNA. These results suggest that confluentin inhibits IMP1-KRAS RNA interaction by binding to the KH1&2 di-domains of IMP1. Since the molecular interaction between IMP1 and its target RNAs is a pre-requisite for the oncogenic function of IMP1, confluentin should be further explored as a potential inhibitor of IMP1 in vivo.

In the boreal forest, open lichen woodlands have been described as an alternative stable state to closed-crown feather moss forest. In this study, we addressed the role of terricolous lichens in stabilizing open woodlands by hindering tree regeneration and/or growth. Based on field and greenhouse experiments, we compared germination and growth of jack pine (Pinus banksiana) on feather mosses (primarily Pleurozium schreberi) and lichens (primarily Cladonia stellaris), using bare mineral soil as a control. Drivers were investigated by (1) manipulating nutrient supply, (2) simulating shade of a closed canopy on the ground layer with the assumption this would mitigate lichen influence on pine growth, and (3) examining pine root ectomycorrhizal colonization and diversity as indicators of pine ability to take up nutrients. Total growth of 6-month-old greenhouse and 2–3-year-old field seedlings, as well as belowground growth of 2-year-old greenhouse seedlings, was significantly greater in moss than in lichen. Seed germination was not affected by ground cover type. Although field phosphorus and base cation availability was greater in mosses than in lichens, fertilization did not entirely compensate for the negative effects of lichens on pine growth in the greenhouse. Ground layer shading had no impact on pine growth. Lichens were associated with reduced abundance and modified composition of the root ectomycorrhizal community. By suggesting that terricolous lichens constitute a less favorable growth substrate than mosses for pine, our results support the hypothesis that lichens contribute to open woodland stability in the potentially closed-crown feather moss forest.

The aim of this study was to investigate the anti-inflammatory activity of a previously un-studied wild mushroom, Echinodontium tinctorium, collected from the forests of north-central British Columbia. The lipopolysaccharide (LPS)-induced RAW264.7 macrophage model was used to study the in vitro anti-inflammatory activity. The crude alkaline extract demonstrated potent anti-inflammatory activity, and was further purified using a &ldquo, bio-activity-guided-purification&rdquo, approach. The size-exclusion and ion-exchange chromatography yielded a water-soluble anti-inflammatory polysaccharide (AIPetinc). AIPetinc has an average molecular weight of 5 kDa, and is a heteroglucan composed of mainly glucose (88.6%) with a small amount of galactose (4.0%), mannose (4.4%), fucose (0.7%), and xylose (2.3%). In in vivo settings, AIPetinc restored the histamine-induced inflammatory event in mouse gluteus maximus muscle, thus confirming its anti-inflammatory activity in an animal model. This study constitutes the first report on the bioactivity of Echinodontium tinctorium, and highlights the potential medicinal benefits of fungi from the wild forests of northern British Columbia. Furthermore, it also reiterates the need to explore natural resources for alternative treatment to modern world diseases.

An immuno-stimulatory polysaccharide (EtISPFa) was purified from water extract of the fungus Echinodontium tinctorium. EtISPFa has an estimated weight average molecular weight (Mw) of 1354 kDa and is composed of glucose (66.2 %), glucuronic acid (10.1 %), mannose (6.7 %), galactose (6.4 %), xylose (5.6 %), rhamnose (3.1 %), fucose (1.8 %), and arabinose (0.2 %). It has multiple glycosidic linkages, with 3-Glcp (19.8 %), 4-GlcpA (10.8 %), 6-Glcp (10.7 %), and 3,6-Glcp (8.7 %) being the most prominent. NMR analysis showed that EtISPFa has a backbone containing mostly of 3-substituted β-glucopyranose with 4-substituted glucopyranosyluronic acid. Short side chains consisting of an average of two β-glycopyranose residues, connected through 1→6 linkages, are attached to the 6-position of about every 4th or 5th backbone glucose residue. EtISPFa is a novel glucuronic acid-containing β-glucan capable of significantly inducing the production of cytokines IL-17, IL-16, MIP-2, G-CSF,GM-CSF, LIF, MIP-1α, MIP-1β, and RANTES in vitro. EtISPFa should be further explored for its immuno-stimulatory activity in vivo.

Wild mushrooms, especially from North America, have not been systematically explored for their medicinal properties. Here we report screening for the growth-inhibitory and immunomodulatory activities of 12 species collected from multiple locations in north-central British Columbia, Canada. Mushrooms were characterized using morphology and DNA sequencing, followed by chemical extraction into 4 fractions using 80% ethanol, 50% methanol, water, and 5% sodium hydroxide. Growth-inhibitory, immunostimulatory, and anti-inflammatory activities of 5 mushrooms (Leucocybe connata, Trichaptum abietinum, Hydnellum sp., Gyromitra esculenta, and Hericium coralloides) are reported here, to our knowledge for the first time. Growth-inhibitory effects were assessed using the cytotoxic MTT assay. Immunostimulatory activity was assessed by tumor necrosis factor-α production in Raw 264.7 macrophages, whereas anti-inflammatory activity was assessed based on the inhibition of lipopolysaccharide-induced tumor necrosis factor-α production. The ethanol and aqueous extracts of Hydnellum sp. were potent growth inhibitors, with a half-maximal inhibitory concentration of 0.6 mg/mL. All 5 fungi displayed strong immunostimulatory activity, whereas only L. connata and T. abietinum showed strong anti-inflammatory activity. For the 7 other fungi investigated, which included well-known medicinal species such as Inonotus obliquus, Phellinus igniarius, and Ganoderma applanatum, the remarkable similarities in the biological activities reported here, and by others for specimens collected elsewhere, suggest that mushrooms can produce similar metabolites regardless of their habitat or ecosystem. This is to our knowledge the first study to explore wild mushrooms from British Columbia for biological activities that are relevant to cancer, and the results provide an initial framework for the selection of mushroom species with the potential for discovery of novel anticancer compounds.

Following a pine beetle epidemic in British Columbia, Canada, we investigated the effect of fire severity on rhizosphere soil chemistry and ectomycorrhizal fungi (ECM) and associated denitrifying and nitrogen (N)-fixing bacteria in the root systems of regenerating lodgepole pine seedlings at two site types (wet and dry) and three fire severities (low, moderate, and high). The site type was found to have a much larger impact on all measurements than fire severity. Wet and dry sites differed significantly for almost all soil properties measured, with higher values identified from wet types, except for pH and percent sand that were greater on dry sites. Fire severity caused few changes in soil chemical status. Generally, bacterial communities differed little, whereas ECM morphotype analysis revealed ectomycorrhizal diversity was lower on dry sites, with a corresponding division in community structure between wet and dry sites. Molecular profiling of the fungal ITS region confirmed these results, with a clear difference in community structure seen between wet and dry sites. The ability of ECM fungi to colonize seedlings growing in both wet and dry soils may positively contribute to subsequent regeneration. We conclude that despite consecutive landscape disturbances (mountain pine beetle infestation followed by wildfire), the “signature” of moisture on chemistry and ECM community structure remained pronounced.

A growth-inhibitory polysaccharide (GIPinv) was purified using size-exclusion and ion-exchange chromatography from the fourth sodium hydroxide extraction step of a fungus found in British Columbia. The fungus was genetically identified as a member of the Paxillus involutus complex. GIPinv has an average molecular weight of 229 kDa and is a heteroglycan composed of glucose (65.9%), galactose (20.8%), mannose (7.8%), fucose (3.2%) and xylose (2.3%). GC–MS methylation analysis suggests that GIPinv has mixed linkages in the backbone containing (1 → 6)-Gal (25.5%), (1 → 4)-Glc (18.3%), (1 → 6)-Glc (8.3%), (1 → 3)-Glc (5.3%) and (1 → 2)-Xyl (4.5%). GIPinv has branching points at (1 → 2, 6)-Man (8.6%) and (1 → 3, 6)-Man (4.9%) having unsubstituted fucose (8.3%) and glucose (16.3%) as terminal sugars. GIPinv had growth-inhibitory activity against several cancer cell lines and triggered apoptosis. GIPinv should be further explored as a potential anti-cancer agent and a unique polysaccharide.

Background: Serpentine ecosystems support different, often unique, plant communities; however, we know little about the soil organisms that associate with these ecosystems. Mycorrhizas, mutualistic symbioses between fungi and roots, are critical to nutrient cycling and energy exchange below ground.Aims: We address three hypotheses: H1, diversity of mycorrhizal fungi in serpentine soils mirrors above-ground plant diversity; H2, the morphology of mycorrhizas and fungi on serpentine soils differs from that on non-serpentine; and H3, mycorrhizal fungal communities of the same or closely related hosts differ between serpentine and non-serpentine soils.Methods: This review focuses on whether plant diversity on serpentine soils correlates with the below ground diversity of mycorrhizal fungi.Results: Studies show that plants and fungi formed abundant ectomycorrhizal and arbuscular mycorrhizal symbioses on and off serpentine soils. No serpentine-endemic fungi were identified. Molecular analyses indicate distinct s...

Trembling aspen (Populus tremuloides) is widely distributed in North American forests. Increased stand productivity with resource availability has been reported, but the relationship between soil microbial community structure and stand productivity remains unclear. To examine soil microbial composition of 4 aspen stand productivity classes, we assessed soil properties, microbial biomass and respiration, and bacterial and ectomycorrhizal diversity. Most variables showed no significant differences between productivity classes. However, mean values for basal respiration (0.05 to 27.99 μg CO2-C·g−1 soil·h−1), bacterial biomass, and metabolic quotient (0.08 to 5.22 CO2-C·mg−1 Cmic·h−1) were lowest in low productivity (Class 1) sites. Bacteria to fungi ratios were significantly lower (P = 0.05) in Class 1 compared to other classes. Microbial biomass ranged from 1.39 to 8.11 mg Cmic·g−1 soil. Thirty-seven distinct aspen ectomycorrhizas (ECM) were characterized, 21 were considered rare (from ≤3 trees). EC...

All members of the Monotropoideae (Ericaceae), including the species, Allotropa virgata and Pleuricospora fimbriolata, are mycoheterotrophs dependent on associated symbiotic fungi and autotrophic plants for their carbon needs. Although the fungal symbionts have been identified for A. virgata and P. fimbriolata, structural details of the fungal-root interactions are lacking. The objective of this study was, therefore, to determine the structural features of these plant root-fungus associations. Root systems of these two species did not develop dense clusters of mycorrhizal roots typical of some monotropoid species, but rather, the underground system was composed of elongated rhizomes with first- and second-order mycorrhizal adventitious roots. Both species developed mantle features typical of monotropoid mycorrhizas, although for A. virgata, mantle development was intermittent along the length of each root. Hartig net hyphae were restricted to the host epidermal cell layer, and fungal pegs formed either along the tangential walls (P. fimbriolata) or radial walls (A. virgata) of epidermal cells. Plant-derived wall ingrowths were associated with each fungal peg, and these resembled transfer cells found in other systems. Although the diffuse nature of the roots of these two plants differs from some members in the Monotropoideae, the structural features place them along with other members of the Monotropoideae in the "monotropoid" category of mycorrhizas.

Ectomycorrhizal (ECM) communities of black spruce (Picea mariana (Mill.) BSP) seedlings were characterized from three habitats spanning a moisture gradient in central British Columbia: black spruce dominated wetlands, black spruce – tamarack wetlands, and black spruce – lodgepole pine uplands. Morphological and molecular (PCR-RFLP) analyses indicated a diverse community of root-associated ECM fungi consisting of 33 morphotypes and 65 genotypes. ECM abundance varied significantly between habitats for six morphotypes. Although many occurred in all three habitats, some occurred in only one or two, and some genotypes had distributions that suggested habitat specificity across the moisture gradient. Intraspecific variation (defined as genotype variation within morphotypes) ranged from one to seven genotypes, depending on ECM morphotype. Both morphological and molecular analyses showed that ECM diversity was greater in upland than in wetland habitats and greater in black spruce – tamarack wetlands than in black spruce dominated wetlands (α ≤ 0.05). Morphological assessment captured contributions to diversity by both abundant and less abundant ECM morphotypes, whereas molecular analysis revealed patterns of genetic variation and habitat distribution at a finer resolution. The study presents the first comprehensive information on black spruce ECM and suggests that ECM community composition and richness varies across the moisture gradient in response to soil heterogeneity and alternate hosts (tamarack and lodgepole pine).

This paper begins with a brief comparison of Franz Kamienski's 1882 view of the fungus-root associations and nutrition of Monotropa hypopitys with our current understanding. The rest of this paper is a re-publication of Shannon Berch's 1985 translation of Kamienski's breakthrough paper in which it was asserted that Monotropa forms a mutualistic symbiosis and is nourished by fungi associated with the roots of neighbouring trees.

Morphological and molecular (polymerase chain reaction – restriction fragment length polymorphism) methods were used to assess ectomycorrhiza (ECM) diversity in naturally regenerating and planted Picea engelmannii Parry ex Engelm. × Picea glauca (Moench) Voss seedlings in two recently clear-cut sites, two clear-cut and broadcast burned sites, and two mature forests (>100 years) in central British Columbia. Based on 24 characterized ECM, burning did not affect overall diversity (Shannon, Simpson, Shannon evenness, and Margalef indices). However, the occurrence and relative abundance of some ECM morphotypes varied significantly. Hebeloma and a Russulaceae type 1 were more abundant and Cenococcum was less abundant in planted seedlings from cut-burned sites compared with those from clear-cut sites. E-strain, MRA, and Amphinema were more abundant in planted seedlings from both cut-burned and clear-cut sites compared with naturally regenerating seedlings from mature sites. ECM diversity of regenerating seedlings was significantly greater in mature forests compared with clear-cut sites and was greater in planted seedlings than naturally regenerating seedlings in clear-cut sites. Molecular analysis of the internal transcribed spacer region of the nuclear-encoded ribosomal RNA gene repeat showed no significant differences among treatments or seedling type. Twenty-two genotypes were identified from eight common morphotypes: Cenococcum (one), Thelephora (two), E-strain, Tuber, Hebeloma, and Russulaceae type 1 (three each), MRA (four), and Amphinema (six). Hebeloma genotypes matched three for Amphinema; genotypes from unidentified lightly colonized tips matched those for E-strain, MRA, and Amphinema-Hebeloma.

Seedlings of Abies grandis (Dougl.) Lindl. (grand fir), Lithocarpus densiflora (Hook. & Arn.) Rehd. (tanoak), Pinus ponderosa Dougl. ex Laws. (ponderosa pine), Pseudotsuga menziesii (Mirb.) Franco (Douglas-fir), and Arbutus menziesii Pursh (madrone) were planted in mixture and monoculture in soil collected from three adjacent forest sites in southwestern Oregon (a clearcut area, a 25-year-old Douglas-fir plantation, and a mature 90- to 160-year-old Douglas-fir - pine forest) to determine the effect of host tree diversity on retrieval of ectomycorrhizal morphotypes. In this greenhouse bioassay, 18 morphotypes of mycorrhizae were recognized overall from all soils with a total of 55 host-fungus combinations: 14 types with ponderosa pine, 14 with Douglas-fir, 10 with tanoak, 10 with grand fir, and 7 for madrone. Four genus-specific morphotypes were retrieved (three on ponderosa pine and one on Douglas-fir), even in mixture situations, demonstrating selectivity of some fungal propagules by their respective host. Five types were detected on all hosts, but not necessarily in soils from all sites. The remaining nine types were associated with two, three, or four hosts, which indicates a wide potential for interspecific hyphal linkages between trees. More morphotypes were retrieved from the monoculture treatments compared with the mixture treatments, although the differences were not significant. Several examples of acropetal replacement of one fungus by another (interpreted as succession) were recorded on all hosts during the course of the experiment. These results illustrate the importance of different host species in maintaining ectomycorrhizal fungus diversity, especially fungi with restricted host range, and the strong potential for fungal linkages between trees in forest ecosystems.Key words: fungal succession, fungal communities, compatibility, Arbutus menziesii, Pseudotsuga menziesii, Pinus ponderosa, Abies grandis, Lithocarpus densiflora.

Laminated-root rot, caused by Phellinus weirii (Murr.) Gilb., is a serious disease affecting Douglas-fir and other commercially important species of conifers in northwestern North America. Recent work has shown that this fungus is successfully reduced or eliminated by the fumigant chloropicrin. However, the effect of this biocide on nontarget organisms, including ectomycorrhizae, is uncertain. Following an initial assessment of organisms 2 years after application of the fumigant, a reassessment of the establishment of ectomycorrhizae on Douglas-fir seedlings was undertaken 4.5 and 5.5 years following chloropicrin application. Our findings show that in areas around stumps treated with 20% and 100% of the labeled dosage and in areas around non-treated stumps, chloropicrin did not adversely affect the formation of ectomycorrhizae on young Douglas-fir seedlings by naturally occurring fungi. No significant effect on the abundance or type of mycorrhizae were detected. On this site, chloropicrin did not affect these mycorrhizal associations, at least for 5 years following application.

The toxic effect of cyclophosphamide on the proliferative cell population of hair follicles plucked from the human scalp was examined by the in vivo nuclear aberration assay. Patients participating in an independent clinical trial received oral low dose cyclophosphamide, intravenous high dose cyclophosphamide or oral placebo treatment. The percent of cells with nuclear aberrations (indicating apoptosis, a special form of cell death) and the percent of mitotic cells, in the hair matrix, were calculated for each patient before treatment and at several time points following cyclophosphamide or placebo treatment. The mean percentages of nuclear aberrations in both the treated Low dose and High dose cyclophosphamide patients were significantly higher than those for the pre-treatment and Placebo patients. The nuclear aberrations in hair follicle cells increased from pre-treatment (and Placebo) to treated Low dose and finally to treated High dose patients. The average percentage for pre-treatment samples from all patients was 0.06 +/- 0.03 SE. For 1 week and 1 month samples from Low dose patients it was 0.35 +/- 0.08 SE, and for combined 2,3 and 4 day samples from High dose patients it was 1.08 +/- 0.12 SE. Cyclophosphamide also had a significant effect on mitosis. A decrease in mitotic activity was observed at 1 month following the initial low dose cyclophosphamide treatment and at 24 +/- 2 h following each of the first two high dose cyclophosphamide treatments. The observed increase in nuclear aberrations following low dose as well as high dose cyclophosphamide suggests that it is feasible to use the nuclear aberration assay for in vivo human genotoxicity testing, using proliferating hair follicle cells.

Pityopus californicus (Eastw.) H. F. Copel., a monotypic member of the Monotropoideae in the family Ericaceae, is a myco-heterotrophic species with distribution limited to the Pacific Northwest of the USA. Young embryos of P. californicus developed mycorrhizal associations in seed packets that had been buried for up to 681 days, suggesting that seeds of P. californicus may require the presence of a fungus to achieve germination. Samples of nongerminated seeds and early stages in embryo and root development were subsequently processed for light microscopy, histochemistry, and transmission electron microscopy (TEM). Nongerminated seeds possessed a thick testa, lacked a shoot and root meristem, and consisted of an embryo with large parenchymatous cells containing protein bodies and starch grains as storage reserves. In the earliest developmental stage (seed coat still attached), fungal hyphae were present on the testa surface and between the testa and embryo. This stage was followed by embryo elongation, the organization of a root apical meristem, and the development of a well-developed fungal mantle surrounding the elongated embryo. At least two morphotypes were identified based on structural characteristics of the mantle. One of these, with ascomycetous septa, had Cenococcum-like features. Late-stage embryo/early root development revealed a typical mantle and Hartig net, with fungal pegs penetrating the outer tangential walls of epidermal cells. Transfer cell-like deposits of wall material, similar to those described in Monotropa spp., enclosed fungal pegs. The development of a Hartig net and fungal pegs suggests that nutrient exchange interfaces are required for seedling development.

Increased numbers of new pathogenic infections and the development of antibiotic resistance to many old pathogens have forced us to focus on prevention, rather than treatment, of infectious diseases. Widespread vaccination has therefore become a major target for many countries. However, numerous countries do not have adequate access to vaccines that are regularly administered in developed countries. As well, there are many pathogens to which there are currently no vaccines available. One of the many new technologies/approaches for addressing these needs is the production of subunit vaccines in transgenic, edible plants, which enable oral vaccine delivery. A variety of plants have now been used as hosts for vaccines, still at the experimental stage, including tobacco, potatoes, carrots, rice, and tomatoes. The main goal of an oral vaccine is the induction of a mucosal immune response and a subsequent systemic immune response. By stimulating a strong mucosal immune response, edible vaccines may provide effective immunity against pathogens that invade both mucosal and systemic sites. Immunization studies with plant-derived subunit vaccines have demonstrated that plants are able to produce recombinant proteins that retain antigenic and immunogenic properties. Although many issues and concerns still remain regarding the safety and efficacy of edible vaccines in humans, current data indicate that edible vaccines are indeed a promising option for the future.

Plant species in the subfamily Monotropoideae are achlorophyllous and have developed a complex mode of nutrition, receiving photosynthates from neighboring trees via shared fungi. To explore the mycorrhizal associations of Monotropa uniflora in central British Columbia (B.C.), plants were sampled from three sites: a Betula-dominated site and two sites with a mixture of conifer and hardwood trees. Fifteen M. uniflora root-clusters were sampled (five per site) and the mycorrhizal diversity was assessed using morphological and molecular (PCR-RFLP analysis and DNA sequencing) methods. Both methods showed that root-clusters (often comprising several hundred mycorrhizal tips) belonging to the same plant appeared to involve fungus monocultures in the family Russulaceae. All mycorrhizae exhibited typical Russula morphology and had mantle cystidia. Two root-clusters, one each from sites 1 and 3, lacked one of the two types of cystidia present on all other root-clusters. PCR-RFLP analysis resulted in three fragment patterns for the 15 root clusters. One molecular fragment pattern included the two root-clusters displaying the single cystidium type plus an additional root-cluster with both cystidia types. DNA sequencing of a portion of the ITS2 region of the ribosomal DNA suggests that the three variants represent different species; two of the variants clustered with the hypogeous fungi Martellia and Gymnomyces. The study provides increased evidence of low diversity and high specificity in the Monotropa-fungus relationship and suggests that M. uniflora associates uniquely with fungi in the family Russulaceae in central B.C.

Thirty-two one-year-old white spruce (Picea glauca (Moench) Voss) seedlings were grown outdoors for one season in 35 L pots buried in the soil. The pots were vertically split in half. One compartment (mineral) was filled with loamy sand. The bottom of the other compartment (organic) was filled with 10 cm sand topped with 15 cm of organic substrates. Two seedling types (16 seedlings each), (i) polystyroblock-grown and (ii) peat-board grown with mechanical root pruning had their root systems split approximately in half into each of the vertical compartments. Controlled-release (26-12-6 N-P-K) fertilizer was added to one or to none (control) of the compartments. Above-ground growth was positively affected by fertilizer placed in either soil compartmnent. Nutrient content of the foliage was greater in fertilized than in unfertilized seedlings; N and P concentrations were significantly increased. Bud reflushing occurred frequently in fertilized seedlings. Unfertilized container-grown seedlings had the fewest roots in either soil compartment. Unfertilized mechanically-pruned seedlings had significantly greater root length, root surface area, and more root tips in mineral than in organic compartments. They also had more P in current-year leaves than did unfertilized container-grown seedlings. Fertilizer added to mineral compartments significantly affected root growth in these compartments only, whereas when added to organic compartments it affected root growth in both compartments. Root systems of the two seedling types were differently affected by fertilizer: in mechanically-pruned seedlings, the number of roots was reduced but their length and diameter increased; in container-grown seedlings, root proliferation was stimulated and this increased total root length and root surface area. Five ectomycorrhizal morphotypes were identified. E-strain was the most abundant. Except for Cenococcum, all morphotypes were present in nursery stock prior to planting. Changes in distribution of morphotypes after planting appeared related to root health condition rather than to applied fertilizer.

This bending magnet beamline has been in operation since February 1995 for the characterization of optical elements (mirrors, gratings, multilayers, detectors, etc.) in the energy range 50–1000 eV. Although it was designed primarily for precision reflectometry of multilayer reflecting optics for EUV projection lithography, it has capabilities for a wide range of measurements. The optics consist of a monochromator, a reflectometer, and refocusing mirrors to provide a small spot on the sample. The monochromator is a very compact, entrance‐slitless, varied‐line‐spacing plane‐grating design in which the mechanically ruled grating operates in the converging light from a spherical mirror working at high demagnification. Aberrations of the mirror are corrected by the line spacing variation, so that the spectral resolving power λ/Δλ is limited by the ALS source size to about 7000. Wavelength is scanned by simple rotation of the grating with a fixed exit slit. The reflectometer has the capability of positioning th...

Fractionation studies were performed of an immune complex-like material measured in rheumatoid synovial fluid using a biological assay based on histamine release from guinea pig lung. This material was eluted from Sepharose 6B in the excluded fraction, from DEAE cellulose predominantly in the third buffer, and from polyacrylamide electrophoresis in a cathodal position. Heterogeneity of the complexes was evident on iso-electric focussing, with one peak of activity at a pH of 4.2--5.6 and the other at pH 6.8--8.6. Purification of the material was limited by a loss of activity which occurred on using three fractionation procedure in sequence. This instability was avoided in the case of Sepharose 6B by adding 0.1% bovine or rabbit albumin to the elution buffer.

A procedure is described for the detailed characterization of multilayer analyzers which can be effectively applied to their design, optimization and application for absolute x-ray spectrometry. An accurate analytical model has been developed that is based upon a simple modification of the dynamical Darwin-Prins theory to extend its application to finite multilayer systems. Its equivalence to the optical E&M solution of the Fresnel equations at each interface is demonstrated by detailed calculation comparisons for the reflectivity of a multilayer throughout the angular range of incidence of 0 to 90° . A special spectrograph and experimental method is described for the measurement of the absolute reflectivity characteristics of the multilayer. The experimental measurements at three photon energies in the 100-2000 eV region are fit by the analytical modified Darwin-Prins equation (MDP) for I( 0), generating a detailed characterization of two "state of the art" multilayers, a sputtered tungsten-carbon of 2d 21 70 A and a molecular lead stearate of 2d 100 A. The fitting parameters that are determined in this procedure are applied to help establish the structural characteristics of the particular multilayer.

A procedure is described for a detailed characterization of multilayer analyzers that can be effectively applied to their design, optimization, and application for absolute x-ray spectrometry in the 100 to 10,000 eV photon energy region. An accurate analytical model has been developed that is based upon a simple modification of the dynamical Darwin-Prins theory to extend its application to finite multilayer systems and to the low energy x-ray region. Its equivalence to the optical E&M solution of the Fresnel equations at each interface is demonstrated by detailed comparisons for the reflectivity of a multilayer throughout the angular range of incidence of 0° to 90°. A special spectrograph and an experimental method are described for the measurement of the absolute reflectivity characteristics of the multilayer. The experimental measurements at three photon energies in the 100 to 2000 eV region are fit by the analytical modified Darwin-Prins equation (MDP) for 1(0), generating a detailed characterization of two state-of-the-art multilayers: sputtered tungsten-carbon with 2d 70 A and a molecular lead stearate with 2d 100 A. The fitting parameters that are determined by this procedure are applied to help establish the structural characteristics of these multilayers.

The rheumatoid biologically active factor (RBAF) was characterized further with respect to its biological and physical characteristics. The histamine-releasing activity of the RBAF in the guinea-pig lung was influenced in the same manner as soluble immune complexes when the lungs were being perfused at 20°C or 45°C or when the perfusate lacked calcium or magnesium or contained N-ethylmaleimide, phenol, theophylline, adrenaline or succinate. The RBAF was consistently associated with complement-fixing activity and RBAF-positive synovial fluid showed a lower total haemolytic complement level than RBAF-negative fluid. However, RBAF activity was not lost following absorption with anti-human beta 1C globulin. There was a higher frequency of free DNA and/or single-stranded DNA in RBAF-positive than negative synovial fluid. RBAF-positive synovial fluid was more active than RBAF-negative fluid in neutrophil chemotaxis when examined at 1:10 but not when undiluted. Mixed IgG–IgM cryoprecipitates failed to show RBAF activity and aggregates seen on analytical ultracentrifugation of rheumatoid synovial fluid did not correspond with the RBAF. The RBAF was stable to freezing and thawing but was labile to acid pH and to heating at 56°C. It was concluded that the RBAF is likely to be a soluble immune complex consisting of IgG and a second constituent which is labile to acid and heat.

By purifying Factor VIII while maintaining physiological concentrations of calcium we have recently demonstrated that about 50% of the procoagulant activity is in a very low molecular weight (VLHW) form not associated with the carrier (VIII: RAG). The remainder is carrier associated and elutes at Vo as a high molecular weight (HMW) compound upon Sepharose 6B chromatography. Reduction of the calcium concentration by increasing the amount of citrate added to heparin results in decreasing the ratio of VLMW:HMW from 1:1 in pure heparin to 1:5 in pure citrate. If citrate is replaced with the more strongly chelating EDTA no VLMW is detectable in the plasma. It has also been found that most of the biochemical purification techniques which have been previously used to prepare Factor VIII for study actually result in the aggregation of this VLMW with the carrier to produce the high molecular weight form. This includes: cryoprecipitation, precipitation by polyethylene glycol and storage -80°C. As well, the VLMW material will self-associate upon freezing to produce an aggregate with a molecular weight of 106. However, this material does not cross-react with rabbit antibody directed against VIII: RAG. The data indicate that many of the previously reported biochemical characteristics, including molecular weights, actually describe species which are artifacts of the isolation process rather than those of the physiologically occuring Factor VIII.