Oral health promotion in the community pharmacy: an evaluation of a pilot oral health promotion intervention.

Introduction Poor oral health is a significant public health concern, costing the NHS in England £3.4 billion annually. Community pharmacies are easily accessible, frequently visited by patients and the community pharmacy contractual framework requires pharmacies to provide healthy living advice to patients - therefore offering a little explored avenue for the delivery of oral health interventions.Methodology A pilot oral health promotion intervention was introduced in five pharmacies in deprived areas of County Durham between September and December 2016. A mixed methods approach to the evaluation was performed, utilising a patient evaluation questionnaire and semi-structured qualitative interviews with pharmacy staff.Results One thousand and eighty-nine participants received the intervention. Following the intervention 72% of participants perceived their knowledge of oral health as much better, 66% definitely intended to change their oral health habits and 64% definitely thought a pharmacy was the right place to receive advice about oral health. Three themes emerged from the qualitative data: (1) intervention feedback, (2) knowledge gap and (3) service development.Discussion The data demonstrated the acceptability of patients to a community pharmacy based oral health intervention, with most patients reporting intentions to change their oral healthcare habits after receiving the intervention. Previous literature has identified a willingness of pharmacy staff to become involved with oral health; this study provides evidence that patients are also receptive to such services being delivered in the community pharmacy setting. Further work is required to assess the benefits of a community pharmacy based oral health intervention and the potential for further growth of this role.Conclusion A community pharmacy is perceived by patients as an acceptable provider of oral health interventions and has the potential to provide positive changes to the oral health of the population.

Influence of Two Acyclic Homoterpenes (Tetranorterpenes) on the Foraging Behavior of Anthonomus grandis Boh.

Previous studies have shown that the boll weevil, Anthonomus grandis, is attracted to constitutive and conspecific herbivore-induced cotton volatiles, preferring the blend emitted by cotton at the reproductive over the vegetative stage. Moreover, this preference was paralleled by the release of the acyclic homoterpenes (tetranorterpenes) (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) and (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene (TMTT) in Delta Opal cotton being higher at the vegetative than at the reproductive stage. Here, we evaluated whether this difference in release of acyclic homoterpenes also occurred in other cotton varieties, and if boll weevils could recognize these compounds as indicators of a specific cotton phenological stage. Results showed that cotton genotypes CNPA TB-90, BRS-293 and Delta Opal all produced higher levels of DMNT and TMTT at the vegetative stage than at the reproductive stage and that these homoterpenes allowed for principal component analysis separation of volatiles produced by the two phenological stages. Electroantennograms confirmed boll weevil antennal responses to DMNT and TMTT. Behavioral assays, using Y-tube olfactometers, showed that adding synthetic homoterpenes to reproductive cotton volatiles (mimicking cotton at the vegetative stage in terms of homoterpene levels) resulted in reduced attraction to boll weevils compared to that to unmodified reproductive cotton. Weevils showed no preference when given a choice between plants at the vegetative stage and the vegetative stage-mimicked plant. Altogether, the results show that DMNT and TMTT are used by boll weevils to distinguish between cotton phenological stages.

Identification of pheromone components and their binding affinity to the odorant binding protein CcapOBP83a-2 of the Mediterranean fruit fly, Ceratitis capitata.

The Mediterranean fruit fly (or medfly), Ceratitis capitata (Wiedemann; Diptera: Tephritidae), is a serious pest of agriculture worldwide, displaying a very wide larval host range with more than 250 different species of fruit and vegetables. Olfaction plays a key role in the invasive potential of this species. Unfortunately, the pheromone communication system of the medfly is complex and still not well established. In this study, we report the isolation of chemicals emitted by sexually mature individuals during the "calling" period and the electrophysiological responses that these compounds elicit on the antennae of male and female flies. Fifteen compounds with electrophysiological activity were isolated and identified in male emissions by gas chromatography coupled to electroantennography (GC-EAG). Within the group of 15 identified compounds, 11 elicited a response in antennae of both sexes, whilst 4 elicited a response only in female antennae. The binding affinity of these compounds, plus 4 additional compounds known to be behaviourally active from other studies, was measured using C. capitata OBP, CcapOBP83a-2. This OBP has a high homology to Drosophila melanogaster OBPs OS-E and OS-F, which are associated with trichoid sensilla and co-expressed with the well-studied Drosophila pheromone binding protein LUSH. The results provide evidence of involvement of CcapOBP83a-2 in the medfly's odorant perception and its wider specificity for (E,E)-α-farnesene, one of the five major compounds in medfly male pheromone emission. This represents the first step in the clarification of the C. capitata and pheromone reception pathway, and a starting point for further studies aimed towards the creation of new powerful attractants or repellents applicable in the actual control strategies.

Sex pheromone communication in two sympatric neotropical stink bug species Chinavia ubica and Chinavia impicticornis.

Chinavia and Nezara spp. stink bugs (Heteroptera: Pentatomidae) include over100 species, with highest diversity in Afrotropical and Neotropical regions. Species thus far studied in these genera utilize trans-(Z)-(4 S)-bisabolene epoxide (BE) and cis-(Z)-(4 S)-BE as major sex pheromone components, with species specificity ensured by different ratios of the two compounds. Gas chromatography (GC) and coupled GC-mass spectrometry (GC-MS) analyses of a volatiles from C. ubica males revealed the presence of two BE isomers in approximately a 90:10 ratio, which were shown by microprobe (1) H NMR to be cis-(Z)-BE and trans-(Z)-BE isomers, respectively. Analyses of volatiles from C. impicticornis males suggested the presence of a single isomer, trans-(Z)-BE, in high purity (>90 %). The absolute configurations of the isomers produced by C. ubica and C. impicticornis were determined using chiral GC analysis (ß-DEX column). Oxidative microchemistry of synthetic standards of cis-(Z)-(4 S)-BE and trans-(Z)-(4R)-BE, and volatiles from male of C. ubica, revealed the absolute stereochemistry of the cis-(Z)-BE to be (1R,2 S,4 S) [cis-(Z)-(4 S) for short]. Similarly, analyses of trans-(Z)-(4 S)-BE and cis-(Z)-(4R)-BE standards, and volatiles from males of C. ubica and C. impicticornis, revealed the absolute stereochemistry of the trans-(Z)-BE to be (1 S,2R,4 S) [trans-(Z)-(4 S) for short]. Olfactometer bioassays with synthetic BEs confirmed attraction of female C. ubica and C. impicticornis to conspecific synthetic pheromone, but not to heterospecific synthetic pheromone. Chinavia impicticornis appeared not to discriminate behaviorally between the conspecific pheromone and its enantiomer. Coupled GC-electroantennography with antennae from females suggested that C. ubica and C. impicticornis possess olfactory receptors for both cis-(Z)-(4 S)-BE and trans-(Z)-(4 S)-BE. The results in this study confirm that C. ubica and C. impicticornis, as for other Chinavia and Nezara spp., utilize cis-(Z)-(4 S)-BE and trans-(Z)-(4 S)-BE as sex pheromone components, with different ratios guaranteeing species specificity. Furthermore, the results suggest that the absolute stereochemistry of BEs may be less important for conspecific recognition than the relative stereochemistry between the epoxide group and the alkyl substituent on the bisabolene ring.

A bioassay for studying behavioural responses of the common bed bug, Cimex lectularius (Hemiptera: Cimicidae) to bed bug-derived volatiles.

The common bed bug, Cimex lectularius (Hemiptera: Cimicidae), has recently re-emerged in increasing numbers, distribution and intensity of infestation in many countries. Current control relies on the application of residual pesticides; but, due to the development of insecticide resistance, there is a need for new tools and techniques. Semiochemicals (behaviour and physiology modifying chemicals) could be exploited for management of bed bugs. However, in order to identify semiochemicals that can be utilised in monitoring or control, a suitable olfactometer is needed that enables the study of the responses of bed bugs to volatile chemicals. Previous studies have used olfactometers that do not separate olfactory responses from responses to physical contact. In this study, a still-air olfactometer was used to measure behavioural responses to different bed bug-derived volatiles presented in an odour pot. Bed bugs were significantly more likely to visit the area above the odour pot first, and more frequently, in the presence of volatiles from bed bug-exposed paper but not in the presence of volatiles from conspecific bed bugs. Bed bug activity was found to be dependent on the presence of the volatiles from bed bug-exposed paper, the time during the scotophase and the sex of the insect being tested. The still-air olfactometer could be used to test putative semiochemicals, which would allow an understanding of their behavioural role in bed bug ecology. Ultimately, this could lead to the identification of new semiochemical tools for bed bug monitoring and control.

Estimating forest LAI profiles and structural parameters using a ground-based laser called 'Echidna'.

There are many techniques for measuring leaf area index (LAI) and forest canopy foliage profiles but their accuracy is questionable. This paper briefly reviews current methods of estimating forest LAI and presents a novel, ground-based laser system, Echidna that can make a wide range of measurements of forest structure, including LAI. Here, use of the system to provide field data and derived gap probabilities in the form of a 'hemispherical photograph with range' is demonstrated. The results show consistency and reproducibility and do not depend on special conditions for the natural light field.

Electrophysiological responses of the lepidopterous stemborers Chilo partellus and Busseola fusca to volatiles from wild and cultivated host plants.

The stemborers Chilo partellus and Busseola fusca are major pests of subsistence cereal farming in Africa. Volatiles released by two cultivated hosts, sorghum and maize (Sorghum bicolor and Zea mays), and two wild grass hosts, Pennisetum purpureum and Hyparrhenia tamba, were collected by air entrainment. Electrophysiologically active components in these samples were detected by coupled gas chromatography-electroantennography (GC-EAG), and the active peaks identified by gas chromatography-mass spectrometry. A total of 41 compounds were identified from the four plant species, all of which, as well as two unidentified compounds, elicited an electrophysiological response from one or both of the stemborers. The compounds included a number of green leaf volatiles and other aliphatic aldehydes, ketones, and esters, mono- and sesquiterpenoids, and some aromatic compounds. EAG studies with authentic samples, conducted at two discriminating doses for all compounds, and dose-response curves for 14 of the most highly EAG-active compounds, showed significant differences in relative responses between species. The compounds that elicited large responses in both species of moths included linalool, acetophenone, and 4-allylanisole, while a number of compounds such as the aliphatic aldehydes octanal, nonanal, and decanal elicited a large response in B. fusca, but a significantly smaller response in C. partellus. Furthermore, the wild hosts produced higher levels of physiologically active compounds compared with either of the cultivated hosts. These differences are discussed in relation to the differential attraction/oviposition of the two stemborers observed in the field and, particularly for eastern African small-scale farming systems, in the context of using a push-pull strategy for their control.

Variation in the load of the horn fly, Haematobia irritans, in cattle herds is determined by the presence or absence of individual heifers.

The distribution of horn flies, Haematobia irritans (L.) (Diptera: Muscidae), in herds of Danish Holstein-Friesian cattle was investigated in two studies conducted during two field seasons. In the first study, highly significant differences in fly distribution between the most and the least fly-susceptible heifers were observed. In one herd, the mean difference between the most fly-susceptible and the most fly-resistant heifers was 268 Ha. irritans specimens. The highest ratio between upper and lower mean fly number was 64.1:1, whereas the lowest was 3.1:1. In the second year, it was demonstrated that the heifers kept their rank in fly attraction over time. The trial clearly demonstrated that some heifers were attracting flies, whereas others, even in the same herd, only carried a few. In the second study, heifers were moved in and out of herds in an attempt to manipulate fly loads in the herds. In year 1, one herd (herd A) received four fly-resistant heifers from another herd (herd B), resulting in a drop in the mean number of flies, whereas herd B received four fly-susceptible heifers from herd A, resulting in an elevation of the mean number of flies. In year 2, a similar pattern emerged using herds C and D, and when the cattle were later returned to their original herds, the fly loads returned to their original distribution. The data presented here show unequivocally that, for horn flies, there can be considerable differences in fly loads for individual heifers within the Holstein-Friesian breed. Furthermore, the overall fly load within herds can be manipulated, and can be reversed. Thus, the distribution in the number of flies within a herd appears to depend on the number of fly-resistant or fly-susceptible heifers. The possible role of chemical factors emitted by heifers, i.e. volatile semiochemicals, in determining differences in fly loads is discussed, whereby attractants are emitted by fly-susceptible heifers and enable flies to locate their host, and repellents are emitted by fly-resistant heifers such that the flies are actively repelled from the herd.

The role of volatile semiochemicals in mediating host location and selection by nuisance and disease-transmitting cattle flies.

The role of volatile semiochemicals in mediating the location and selection within herds of Holstein-Friesian heifers by nuisance and disease-transmitting cattle flies was investigated using coupled gas chromatography-electrophysiology (GC-EAG), coupled gas chromatography-mass spectrometry (GC-MS), electrophysiology (EAG), laboratory behaviour and field studies. Using volatile extracts collected by air entrainment from heifers in the Netherlands, a number of active peaks were located by coupled GC-EAG for Musca autumnalis (de Geer) (Diptera: Muscidae) and Haematobia irritans (L.) (Diptera: Muscidae). Volatile samples were also collected from two heifers in Denmark shown in previous counting experiments to differ significantly in their fly loads. Coupled GC-EAG using Ha. irritans antennae revealed differences in the EAG response to the samples, with additional EAG activity in the sample collected from the heifer with the lower fly load. To identify more EAG active compounds, volatiles were also collected from 48-h-old urine by air entrainment. In total, 23 compounds were located and identified by coupled GC-EAG and GC-MS. Further electrophysiological testing of these compounds with five fly species [M. autumnalis, Ha. irritans, Hydrotaea irritans (L.) (Diptera: Muscidae), Stomoxys calcitrans (L.) (Diptera: Musicidae) and Wohlfahrtia magnifica (Schiner) (Diptera: Sarcophagidae)] showed that only some of the compounds were physiologically active across the range of flies tested. These included 1-octen-3-ol, 6-methyl-5-hepten-2-one, (Z)-3-hexen-1-ol, naphthalene, and all EAG active compounds identified from urine. Compounds showing significant EAG activity were tested for behavioural activity using a wind-tunnel designed for measuring upwind flight behaviour. At certain concentrations, 1-octen-3-ol, 6-methyl-5-hepten-2-one and 3-octanol increased upwind flight, whereas naphthalene, propyl butanoate and linalool reduced upwind flight. In field studies using small herds of heifers ranked according to their fly load, individual slow-release formulations of 1-octen-3-ol and 6-methyl-5-hepten-2-one, when applied to low and high fly loading heifers, reduced fly loads on these individuals. This study provides evidence for the hypothesis that the natural differential attractiveness within herds of Holstein-Freisian heifers, i.e. a single host species, for cattle flies is partly due to differences in volatile semiochemicals emitted from the host. It is suggested that this phenomenon applies to other vertebrate host species and their associated insect pests.

Plant stress signalling: understanding and exploiting plant-plant interactions.

When plants are attacked by insects, volatile chemical signals can be released, not only from the damaged parts, but also systemically from other parts of the plant and this continues after cessation of feeding by the insect. These signals are perceived by olfactory sensory mechanisms in both the herbivorous insects and their parasites. Molecular structures involved can be characterized by means of electrophysiological assays, using the insect sensory system linked to chemical analysis. Evidence is mounting that such signals can also affect neighbouring intact plants, which initiate defence by the induction of further signalling systems, such as those that increase parasitoid foraging. Furthermore, insect electrophysiology can be used in the identification of plant compounds having effects on the plants themselves. It has been found recently that certain plants can release stress signals even when undamaged, and that these can cause defence responses in intact plants. These discoveries provide the basis for new crop protection strategies, that are either delivered by genetic modification of plants or by conventionally produced plants to which the signal is externally applied. Delivery can also be made by means of mixed seed strategies in which the provoking and recipient plants are grown together. Related signalling discoveries within the rhizosphere seem set to extend these approaches into new ways of controlling weeds, by exploiting the elusive potential of allelopathy, but through signalling rather than by direct physiological effects.

Characterization of (1'R,4S,4aR,7S,7aR)-dihydronepetalactol as a semiochemical for lacewings, including Chrysopa spp. and Peyerimhoffina gracilis.

The enantiomerically pure diastereoisomers (1R,4S,4aR,7S,7aR)- (1) and (1R,4R,4aR,7S,7aR)-dihydronepetalactol (2) were synthesized diastereoselectively from a renewable resource, (4aS,7S,7aR)-nepetalactone (3), isolated as the main constituent of the essential oil of the catmint plant Nepeta cataria. The stereochemistry of the compounds was determined by NMR spectroscopy and X-ray crystallography, and the compounds were identified, respectively, as neomatatabiol and isoneomatatabiol, natural products from Actinidia polygama, for which the lactol stereochemistry was previously incompletely defined. Compound 1 was found to catch significant numbers of three species of lacewing in the field: in Korea. Chrysopa cognata, and in the United Kingdom, Nineta vittata and most notably Peverimhoffina gracilis. All species caught in significant numbers were found more frequently in traps releasing 1 than 2, while more C. cognata, C. formosa, and C. phyllochroma were found in traps releasing (1R,4aS,7S,7aR)-nepetalactol (4). The catch of P. gracilis with 1 is of particular interest as this lacewing has only recently been recorded in the United Kingdom. Where sexed, the lacewings of all species trapped were found to be male, implying a possible pheromonal role for these or structurally related compounds.

Retrospective study of the association of stress and smoking during pregnancy in rural women.

Women who continue to smoke during pregnancy put themselves and their fetuses at serious risk for complications. Various smoking cessation programs have been designed that specifically target pregnant smokers. Longitudinal studies, however, have shown that there is a group of women who are unable to quit smoking while pregnant. Women from a rural area of the Mid-West (N=299) were interviewed postpartum to determine the stresses these women experienced prenatally and the association of the stress with continuing to smoke during pregnancy. Subjects were divided into three groups: Nonsmokers, Quitters, Smokers. This study not only confirms other reports that these women are more stressed but also documents some of the major stressors. Statistically significant differences were found between groups for financial worries (P=.0002), problems with the family (P<.001), and domestic violence (P<.001). Assessing pregnant women for stress and, especially, domestic violence should be part of the implementation of the Clinical Practice Guidelines for Smoking Cessation.

Response of the ladybird parasitoid Dinocampus coccinellae to toxic alkaloids from the seven-spot ladybird, Coccinella septempunctata.

Electrophysiological and behavioral responses of the ladybird parasitoid Dinocampus coccinellae to volatiles from the seven-spot ladybird, Coccinella septempunctata, were investigated to identify semiochemicals involved in host location. Coupled gas chromatography-electroantennography (GC-EAG) with D. coccinellae located a small peak of prominent activity in an extract of volatiles from adult C. septempunctata. The active compound was identified by coupled GC-mass spectrometry and by comparison with an authentic sample as the free-base alkaloid precoccinelline, which forms part of the toxic defense of this ladybird. Behavioral studies in an olfactometer showed that D. coccinellae was significantly attracted to the volatile extract and also to the alkaloid. Myrrhine, a stereoisomer of precoccinelline found in low amounts in C. septempunctata and in other ladybird species, was shown to be electrophysiologically active and significantly attractive. Perception of ladybird alkaloids by D. coccinellae is a rare example of toxicants acting as aerially transmitted cues for interactions between the third and fourth trophic levels.

Higher-order structure of chromatin and chromosomes.

The linear array of nucleosomes that comprises the primary structure of chromatin is folded and condensed to varying degrees in nuclei and chromosomes forming 'higher order structures'. We discuss the recent findings from novel experimental approaches that have yielded significant new information on the different hierarchical levels of chromatin folding and their functional significance.

New roles for cis-jasmone as an insect semiochemical and in plant defense.

cis-jasmone, or (Z)-jasmone, is well known as a component of plant volatiles, and its release can be induced by damage, for example during insect herbivory. Using the olfactory system of the lettuce aphid to investigate volatiles from plants avoided by this insect, (Z)-jasmone was found to be electrophysiologically active and also to be repellent in laboratory choice tests. In field studies, repellency from traps was demonstrated for the damson-hop aphid, and with cereal aphids numbers were reduced in plots of winter wheat treated with (Z)-jasmone. In contrast, attractant activity was found in laboratory and wind tunnel tests for insects acting antagonistically to aphids, namely the seven-spot ladybird and an aphid parasitoid. When applied in the vapor phase to intact bean plants, (Z)-jasmone induced the production of volatile compounds, including the monoterpene (E)-beta-ocimene, which affect plant defense, for example by stimulating the activity of parasitic insects. These plants were more attractive to the aphid parasitoid in the wind tunnel when tested 48 h after exposure to (Z)-jasmone had ceased. This possible signaling role of (Z)-jasmone is qualitatively different from that of the biosynthetically related methyl jasmonate and gives a long-lasting effect after removal of the stimulus. Differential display was used to compare mRNA populations in bean leaves exposed to the vapor of (Z)-jasmone and methyl jasmonate. One differentially displayed fragment was cloned and shown by Northern blotting to be up-regulated in leaf tissue by (Z)-jasmone. This sequence was identified by homology as being derived from a gene encoding an alpha-tubulin isoform.

Plant signalling and induced defence in insect attack.

Abstract Plants can produce compounds which act as semiochemicals, that is, signals modifying the development or behaviour of other organisms without having direct physiological activity. Among such semiochemicals are plant stress signals associated with the induction of defence systems, and these may include phytopheromones that naturally influence plant development. It is well known that plant-derived semiochemicals can be exploited by colonizing organisms, particularly pathogens and insect pests. Recently proposed external signals not yet proven as natural phytopheromones are nitric oxide and the volatile methyl esters of jasmonic and salicylic acids. Since it is now possible to use sophisticated electrophysiological techniques to investigate insect interactions with prospective phytopheromones, the detection and characterization of signalling systems has been made much easier and can provide a molecular characterization of signals that are active beyond the insects themselves. In addition to the advances these studies have brought to the understanding of plant/insect and plant/plant interactions, plant signals are potentially valuable in the regulation of gene expression for improved or alternative approaches to crop protection or for other developmental processes in plants.

Aphids, predators and parasitoids.

A number of studies have demonstrated the role of herbivore-induced release of plant volatiles in mediating foraging behaviour of aphid parasitoids, particularly with the parasitoid Aphidius ervi, its aphid host Acyrthosiphon pisum and the aphid food plant Vicia faba. These studies have shown that feeding by the aphid alters the composition of volatiles released by the plant and that these compounds act as synomones for the foraging parasitoid. Of particular interest is the species-specificity of the herbivore-induced synomones associated with different aphids feeding on V. faba. Aphids employ various pheromones that mediate behaviour, particularly mating and alarm responses. These pheromones play important roles in reproduction and defence against predation and parasitism. Many species of aphids reproduce sexually on their primary hosts during the autumn and the sexual females produce a pheromone that attracts males. The sex pheromones for a number of aphid species have been identified and laboratory and field studies have shown that synthetic material can act as a kairomone in attracting predators and parasitoids. The aphid alarm pheromone is released from the cornicles of aphids when they are attacked by predators or parasitoids. The activity of the main alarm pheromone component, (E)-beta-farnesene, is inhibited by the related sesquiterpene hydrocarbon beta-caryophyllene, which is reported to attract the lacewing Chrysoperla carnea. In addition, electrophysiological studies have shown that the seven-spot ladybird, Coccinella septempunctata, possesses specific olfactory receptors for (E)-beta-farnesene and beta-caryophyllene. Laboratory studies show these compounds to have behavioural activity with C. septempunctata, suggesting that they may be involved in prey location.

Exploiting insect responses in identifying plant signals.

Plants produce compounds which can act as semiochemicals, i.e. as signals modifying the behaviour and development of other organisms without having direct physiological activity, and which generally have extremely benign toxicological properties. Such semiochemicals include plant stress signals and possibly also phytopheromones associated with the induction of defence systems but which may also be exploited by colonizing organisms. Since it is now possible to use sophisticated electrophysiological techniques to investigate insect interactions with phytopheromones, the detection and characterization of such signalling systems is greatly facilitated, particularly when combined with behavioural techniques for confirming the biological relevance of the signals. As well as the contribution of these studies to the understanding of plant-insect and plant-plant interactions, plant signals are potentially valuable in the regulation of gene expression for improved or alternative approaches to crop protection. It would also be possible to use these signals to 'switch on', when required, other genes associated with beneficial agronomic or nutritional crop traits.

The nature of the nucleosomal barrier to transcription: direct observation of paused intermediates by electron cryomicroscopy.

Transcribing SP6 RNA polymerase was arrested at unique positions in the nucleosome core, and the complexes were analyzed using biochemical methods and electron cryomicroscopy. As the polymerase enters the nucleosome, it disrupts DNA-histone interactions behind and up to approximately 20 bp ahead of the elongation complex. After the polymerase proceeds 30-40 bp into the nucleosome, two intermediates are observed. In one, only the DNA ahead of the polymerase reassociates with the octamer. In the other, DNA both ahead of and behind the enzyme reassociates. These intermediates present a barrier to elongation. When the polymerase approaches the nucleosome dyad, it displaces the octamer, which is transferred to promoter-proximal DNA.