A set of simple decision matrices for prioritizing collection of rare plant species for ex situ conservation

Because it is virtually impossible to collect seed or tissue for ex situ conservation banks from every known population of rare plant species, it is important to rank populations systematically in terms of their priority for collection. The New England Wild Flower Society, which maintains a regional seed bank, developed a set of three complementary decision matrices in spreadsheet form by which to prioritize all occurrences of all state-listed rare plant species in New England in terms of their urgency and feasibility for collection. Data on 4333 occurrences, spanning 456 taxa, were collated from six state Natural Heritage Programs. The first decision matrix ranked taxa in terms of their amenability to storage or propagation at ex situ institutions, and determined whether any known New England occurrences were reproductive. The second matrix further ranked taxa in terms of their global and regional rarity and the viability and genetic and geographic representation of collections already present in the bank. The third matrix scored individual occurrences within each taxon in terms of the presence of imminent threat, reproductive status, vigor, protection status, potential genetic distance from other occurrences, availability of landowner permission, and their current status in the bank. Occurrences were then sorted in ascending order by total matrix score; those with low scores were at the top of the list for collection priority. 3743 occurrences were deemed eligible for collection. Scores ranged from 14.5 to 182, and were influenced most strongly by the number of occurrences per taxon. Clear breakpoints were apparent in the distribution of scores, with clusters of uncommon taxa at the low end of the scale and a long tail created by taxa with more numerous occurrences in New England. These breakpoints could potentially be used to prioritize groups of occurrences that should receive the first attention for collection, while postponing collection of higher-scoring groups. Fewer than 1% of occurrences were misclassified, according to post hoc inspection. This simple set of decision matrices can be adapted by a wide range of institutions involved in ex situ conservation.     

Identifying spatio-temporal patterns of transboundary disease spread: examples using avian influenza H5N1 outbreaks

Characterizing spatio-temporal patterns among epidemics in which the mechanism of spread is uncertain is important for generating disease spread hypotheses, which may in turn inform disease control and prevention strategies. Using a dataset representing three phases of highly pathogenic avian influenza H5N1 outbreaks in village poultry in Romania, 2005–2006, spatio-temporal patterns were characterized. We first fit a set of hierarchical Bayesian models that quantified changes in the spatio-temporal relative risk for each of the 23 affected counties. We then modeled spatial synchrony in each of the three epidemic phases using non-parametric covariance functions and Thin Plate Spline regression models. We found clear differences in the spatio-temporal patterns among the epidemic phases (local versus regional correlated processes), which may indicate differing spread mechanisms (for example wild bird versus human-mediated). Elucidating these patterns allowed us to postulate that a shift in the primary mechanism of disease spread may have taken place between the second and third phases of this epidemic. Information generated by such analyses could assist affected countries in determining the most appropriate control programs to implement, and to allocate appropriate resources to preventing contact between domestic poultry and wild birds versus enforcing bans on poultry movements and quarantine. The methods used in this study could be applied in many different situations to analyze transboundary disease data in which only location and time of occurrence data are reported.     

Black cohosh (Cimicifuga racemosa L.) protects against menadione-induced DNA damage through scavenging of reactive oxygen species: bioassay-directed isolation and characterization of active principles

The roots/rhizomes of Cimicifuga racemosa L. (Nutt.) (black cohosh) have traditionally been used to treat menopausal symptoms through an unknown mechanism of action. In an effort to determine if black cohosh had additional health benefits, methanol extracts were investigated for their potential to scavenge reactive oxygen species and to protect against menadione-induced DNA damage. These extracts effectively scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals. In addition, the extracts showed dose-dependent decreases in DNA single-strand breaks and oxidized bases induced by the quinone menadione using the comet (single-cell gel electrophoresis assay) and fragment length associated repair enzyme assays, respectively. Bioassay-directed fractionation of the methanolic extracts using the DPPH assay as a monitor led to the isolation of nine antioxidant active compounds: caffeic acid (1), methyl caffeate (2), ferulic acid (3), isoferulic acid (4), fukinolic acid (5), cimicifugic acid A (6), cimicifugic acid B (7), cimicifugic acid F (8), cimiracemate A (9), and cimiracemate B (10). Six of these antioxidants were found to reduce menadione-induced DNA damage in cultured S30 breast cancer cells with the following order of potency: methyl caffeate (2) > caffeic acid (1) > ferulic acid (3) > cimiracemate A (9) > cimiracemate B (10) > fukinolic acid (5). These data suggest that black cohosh can protect against cellular DNA damage caused by reactive oxygen species by acting as antioxidants.     

Plant life history traits of rare versus frequent plant taxa of sandplains: Implications for research and management trials

I apply a comparative, functional group approach to coastal sandplain grassland taxa in order to examine whether rare plant species share certain aspects of rarity and life history characters that are distinct from their more common, co-occurring congeners in these habitats. I compiled a comparative data set containing 16 variables describing biogeographic distributions, level of imperilment, habitat specialization, vegetative versus sexual reproduction, seed dispersal, and dormancy of 27 closely-related pairs of plant species that contrast in their abundance (infrequent versus common) in coastal sandplain grasslands. Frequent and infrequent species were paired within genera (or closely related genera) and thus distributed equivalently across families to control for phylogenetic bias. Paired comparisons revealed that infrequent species were intrinsically rarer range-wide, and exhibited a narrower range and more habitat specialization than their common relatives. A classification tree distinguished infrequent species from common species on the basis of higher habitat specialization, larger seed size, smaller plant height, less reliance less on vegetative (colonial) reproduction, and tendency toward annual or biennial life history. Research and management steps to reduce competition from larger-satured, colonial, perennial species are recommended for these infrequent species. Basic research involving more species and more data on ecophysiological characters, demography, and competitive interactions are needed to identify critical life history traits that will influence responses to particular management regimes.     

Discussion of a well-designed clinical trial which did not demonstrate effectiveness: UIC center for botanical dietary supplements research study of black cohosh and red clover

The performance of a clinical trial for pharmaceutical agents is usually undertaken only after there is likely benefit demonstrated from the use of the putative agent. The consideration of botanical products as pharmaceutical agents must similarly go through a rigorous evaluation process. The present work reviews the recently published phase II study evaluating the effectiveness of black cohosh and red clover in a randomized trial with conjugated equine estradiol/medroxyprogesterone acetate and placebo for the treatment of menopausal symptoms. We analyze the possible reasons why this study failed to show benefit for either botanical product in reducing menopause-related vasomotor symptoms.     

Inhibition of insect cytochrome P-450 by some metyrapone analogues and compounds containing a cyclopropylamine moiety and their evaluation as inhibitors of juvenile hormone biosynthesis

Two metyrapone analogues, 2-(l-imidazolyl)-2-methyl-l-phenyl-l-pro-panone (A-phenyl-B-imidazolyl-metyrapone; III) and 2-methyl-l-phenyl-2-{1,2,4-triazol-l-yl)-l -propanone (A-phenyl-B-triazolyl-meiyrapone; IV) as well as two cyclopropylamine derivatives. N-cyclopropyl-4-icrt-butylbenzylamine (V) and N-cyclopropyl-4-(3,7-dimethyl-7-methoxy-octyloxy)benzamide (cyclopropylamine acylated with a JH analogue acid of known structure; VI) were synthesized and evaluated in biological assays for JH biosynthesis on cockroach, Diploptera punctata corpora allata and egg growth in adult cockroach as well as for mixed function oxidase activities, i.e. epoxidation of aldrin to dieldrin and O-demethylation of 7-methoxy-4-methylcoumarin to 7-hydroxy-4-methylcoumarin on microsomes from housefly, Musca domestica, abdomen and from cockroach midgut. Compound VI was a good in-vitro inhibitor of JH biosynthesis, but it had significantly lower activities in the assays for inhibition of microsomal cytochrome P-450. Compound IV and metyrapone had moderate activity as inhibitors of oocyte growth. Compounds III, IV and V were more potent inhibitors of housefly aldrin epoxidation than metyrapone and they inhibited the enzyme activity by almost 100% at 02mM, while in cockroach midgut microsome assay metyrapone was more potent than these three compounds.     

Spatial analysis and incidence–density relationships for downy mildew on hop

The spatial pattern of downy mildew (Pseudoperonospora humuli) on hop (Humulus lupulus) was characterized over 4 years to aid in deriving an appropriate incidence–density relationship. From 472 disease assessments (datasets), discrete distributions were fitted to the datasets to determine aggregation of disease density. Where distributions were able to be fitted, the Poisson distribution fitted 4% of the datasets and the negative binomial distribution fitted 87% of the datasets. Larger‐scale patterns of disease were assessed by autocorrelation and runs analysis; both indicated aggregation of diseased plants was less common than aggregation of disease within plants. Taylor’s power law indicated disease density was aggregated and related to mean disease density in all years. Disease incidence and density were linked by saturation‐type relationships based on the zero term of the negative binomial distribution or an empirical regression. Certain individual datasets were not described well by any incidence–density model, particularly when disease density was greater than about 0·8 diseased shoots per plant with the cultivar Cascade. When applied to 56 validation datasets, 88% of the variation in observed disease incidence was explained by the incidence–density models. Under conditions where sampling would be implemented for disease management, the requisite conditions appear to be in place for a binomial sampling plan for downy mildew.     

Fibrin–alginate hydrogel supports steroidogenesis,in vitro maturation of oocytes and parthenotes production from caprine preantral follicles cultured in group

This study aimed to establish a culture system that improves the in vitro development of caprine preantral follicles. In a first experiment, follicles were encapsulated as a single unit per bead and cultured singly or in groups or with five follicles in the same alginate (ALG) bead for 18 days. In a subsequent experiment, the “five follicles per bead” design was chosen to culture in ALG, fibrin–alginate (FA) or hyaluronate (HA) for 18 days. In a third experiment, we chose the five follicles per bead in FA to culture for 30 days. The culture set‐up of five follicles per ALG bead increased antrum formation and follicle diameter compared to the other culture designs (p < .05). Moreover, under this condition, 44.44% of the oocytes from in vitro cultured preantral follicles reached meiotic resumption. A significant increase of follicle diameter occurred in attachment system and FA (p < .05), but the ALG condition reached the highest among all groups on day 18 (p < .05). Follicles encapsulated in matrix produced more estradiol and progesterone than attachment system (p < .05). The expression of MMP‐9 mRNA was higher in FA than in other groups (p < .05) and similar to antral follicles from in vivo control (p > .05). Only FA group resulted in oocytes matured. After 30 days, oocytes from preantral follicles in vitro grown in FA developed to eight‐cell parthenotes. In conclusion, a culture system using FA supported the development of caprine preantral follicles cultured in group and included in the same bead of hydrogel, improving the oocyte maturation and producing parthenotes.     

Cytochrome P-450 in insects: 4. Reconstitution of cytochrome P-450-dependent monooxygenase activity in the house fly

Two cytochrome P-450-containing fractions were isolated from detergent-solubilized house fly microsomes by hydrophobic chromatography on a tryptamine-Sepharose gel. These fractions (designated P-450-1 and P-450-2) were distinctive in their spectral characteristics and in their profiles following electrophoresis in the presence of sodium dodecyl sulfate. Both fractions exhibited NADPH-dependent epoxidase activity when reconstituted with purified house fly cytochrome P-450 reductase and phospholipid. The aldrin epoxidase activity of fraction P-450-1 was twice that of P-450-2 even though heptachlor epoxidase activity of the fractions was equivalent. O-Demethylase activity with 7-methoxy-4-methylcoumarin was detectable only in the P-450-2 fraction.