Intact Bodies Research Articles

Effect of Varieties/Hybrids of Cotton on the Parasitization Efficiency of Trichogramma chilonis Ishii

Six cultivars, namely, LD 327, LH 1556, Ganga Kaveri-151, Ankur 651, Dbaval-2 and LHH 144 were e,'aluated for testing efficacy of Trichogramma chilonis Ishii. T. chilonis was released 13 times during July to September at weekly interval @ 1,50,000 per ha. Four sprays of compatible insecticides were given during this period. The lowest bollworm incidence among intact fruiting bodies was recorded in LD327 (1.40%) which was on par with Ankur 651 (2.06%) and LHH144 (2.10%) but significantly lower than other cultivars. There was no significant difference in mean bollworm incidence among green bolls, parasitization, and seed cotton yield in various cultivars.

Microglia-associated granule cell death in the normal adult dentate gyrus

Microglial cells are constantly monitoring the central nervous system for sick or dying cells and pathogens. Previous studies showed that the microglial cells in the dentate gyrus have a heterogeneous morphology with multipolar cells in the hilus and fusiform cells apposed to the granule cell layer both at the hilar and at the molecular layer borders. Although previous studies showed that the microglia in the dentate gyrus were not activated, the data in the present study show dying granule cells apposed by Iba1-immunolabeled microglial cell bodies and their processes both at hilar and at molecular layer borders of the granule cell layer. Initially, these Iba1-labeled microglial cells surround individual, intact granule cell bodies. When small openings in the plasma membrane of granule cells are observed, microglial cells are apposed to these openings. When larger openings in the plasma membrane occur at this site of apposition, the granule cells display watery perikaryal cytoplasm, watery nucleoplasm and damaged organelles. Such morphological features are characteristic of neuronal edema. The data also show that following this localized disintegration of the granule cell’s plasma membrane, the Iba1-labeled microglial cell body is found within the electron-lucent perikaryal cytoplasm of the granule cell, where it is adjacent to the granule cell’s nucleus which is deformed. We propose that granule cells are dying by a novel microglia-associated mechanism that involves lysis of their plasma membranes followed by neuronal edema and nuclear phagocytosis. Based on the morphological evidence, this type of cell death differs from either apoptosis or necrosis.

Microglia Activated with the Toll-Like Receptor 9 Ligand CpG Attenuate Oligomeric Amyloid β Neurotoxicity in in Vitro and in Vivo Models of Alzheimer’s Disease

Soluble oligomeric amyloid beta (oAbeta) 1-42 causes synaptic dysfunction and neuronal injury in Alzheimer's disease (AD). Although accumulation of microglia around senile plaques is a hallmark of AD pathology, the role of microglia in oAbeta1-42 neurotoxicity is not fully understood. Here, we showed that oAbeta but not fibrillar Abeta was neurotoxic, and microglia activated with unmethylated DNA CpG motif (CpG), a ligand for Toll-like receptor 9, attenuated oAbeta1-42 neurotoxicity in primary neuron-microglia co-cultures. CpG enhanced microglial clearance of oAbeta1-42 and induced higher levels of the antioxidant enzyme heme oxygenase-1 in microglia without producing neurotoxic molecules such as nitric oxide and glutamate. Among subclasses of CpGs, class B and class C activated microglia to promote neuroprotection. Moreover, intracerebroventricular administration of CpG ameliorated both the cognitive impairments induced by oAbeta1-42 and the impairment of associative learning in Tg2576 mouse model of AD. We propose that CpG may be an effective therapeutic strategy for limiting oAbeta1-42 neurotoxicity in AD.

Mechanisms of Aqueous Extraction of Soybean Oil

Aqueous extraction processing (AEP) of soy is a promising green alternative to hexane extraction processing. To improve AEP oil yields, experiments were conducted to probe the mechanisms of oil release. Microscopy of extruded soy before and after extraction with and without protease indicated that unextracted oil is sequestered in an insoluble matrix of denatured protein and is released by proteolytic digestion of this matrix. In flour from flake, unextracted oil is contained as intact oil bodies in undisrupted cells, or as coalesced oil droplets too large to pass out of the disrupted cellular matrix. Our results suggest that emulsification is an important extraction mechanism that reduces the size of these droplets and increases yield. Protease and SDS were both successful in increasing extraction yields. We propose that this is because they disrupt a viscoelastic protein film at the droplet interface, facilitating droplet disruption. An extraction model based on oil droplet coalescence and the formation of a viscoelastic film was able to fit kinetic extraction data well.

Aqueous Extraction of Oil Bodies from Maize Germ (Zea mays) and Characterization of the Resulting Natural Oil-in-Water Emulsion

Oil bodies in the form of an adequately dispersed suspension were extracted from maize germ by applying aqueous extraction. The effect on the yield of oil body extraction of parameters, such as pH of aqueous medium, state of germ comminution, and number of successive extraction steps applied, was evaluated, indicating that an extraction yield as high as 95% could be reached when a finely comminuted germ material is extracted 3 times under alkaline conditions. The extracted oil body suspension was coagulated by pH manipulation, and the resulting cream, consisting of intact oil bodies, was studied with respect to protein composition, particle size distribution, and oil body zeta potential. Changes in particle mean diameter resulting during oil body cream storage as well as the creaming behavior of emulsions prepared by cream dilution with water were also investigated. The findings are discussed in terms of the presence at the oil body surface of an adsorbed mixed layer made up of phospholipids, oleosins, and extraneous germ proteins that determine the physical stability of oil droplets upon long-term storage.

Infectious agents and lymphoma development: molecular and clinical aspects

This review is focused on the role of infectious agents in the development of some lymphoma entities. Associations involving bacterial infections mostly regard marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT)-type. Some paradigmatic examples of these associations include the Helicobacter pylori-related gastric MALT lymphoma and the more recently reported links between Chlamydophila psittaci and ocular adnexal MALT lymphomas and Borrelia burgdorferi and cutaneous MALT lymphomas. The well-documented association between Epstein-Barr virus infection and related lymphoproliferative disorders are analysed as an example of lymphotropic virus with tumourigenic activity. Molecular, biological and clinical features as well as therapeutic implications of these associations are analysed and future perspectives in this field are discussed.

Environmental and pheromonal control of precopulatory behavior for synchronized mating in the white grub beetle, Dasylepida ishigakiensis (Coleoptera: Scarabaeidae)

Precopulatory behavior of the white grub beetle, Dasylepida ishigakiensis, was observed under laboratory conditions to determine the environmental factors controlling mating behavior, which is synchronized among individuals at dusk in the field. When light intensity was gradually decreased, both females and males sequentially started moving their antennae, legs and abdomen, then walking, and females rhythmically extruded their abdomen while males opened the elytra to fly. Pheromone release by females was suggested in a wind tunnel assay in which males oriented themselves to both intact and crushed bodies of females, but not of males. When illumination was suddenly turned off, precopulatory behavior of females and males was similarly induced but the frequencies of calling and flying behaviors were significantly reduced as compared with those observed when light intensity was gradually reduced. This result may suggest that gradual darkening is an important factor for their synchronized appearance from the soil and subsequent mating behaviors, including pheromone release in females and orientation flight in males. These behaviors were significantly suppressed at low temperatures below 18°C. This result supports our previous conclusion that temperature is the primary factor controlling the emergence and mating activity of sexually mature beetles in the field.

Nitrogen mustard (Chlorambucil) has a negative influence on early vascular development

The sulphur and nitrogen mustards are strong alkylating agents, which induces inflammations of the skin including blistering right up to ulcerations. Depending on the severity, the wounds may need weeks to heal. In the past it was shown that sulphur mustard has a destructive effect on endothelial precursor cells, which have been shown to play a pivotal role in the wound healing reaction by inducing neovascularisation. However, for these alkylating agents as well as for sulphur mustard nothing is known about their effects on endothelial precursors. Therefore, we investigated and compared the influence of Chlorambucil on proliferation, apoptosis and differentiation of endothelial cells in intact mouse embryoid bodies (EB). EBs were treated at different developmental stages and with different periods of Chlorambucil treatment. It was found that in each developmental stage and under each treatment period's Chlorambucil has an extremely negative effect on the vascularisation with a vessel reduction of around 99%. Of particular importance was the negative effect of treatment around day 3 of the development. On this day we found 377 vessels under control conditions but only 1.6 vessels under 24 h treatment of Chlorambucil. At this point in time many endothelial precursors can be found in the EB. Moreover, a negative effect on all stem cells was evident at this point in time, shown by an extreme reduction in EB size with 17.9 mm 2 for the control and only 1.55 mm 2 under Chlorambucil treatment. This negative effect on the vascularisation, on endothelial precursors but also on stem cells in general is of possible importance for impaired wound healing.

Essential role of BMPs in FGF-induced secondary lens fiber differentiation

It is widely accepted that vitreous humor-derived FGFs are required for the differentiation of anterior lens epithelial cells into crystallin-rich fibers. We show that BMP2, 4, and 7 can induce the expression of markers of fiber differentiation in primary lens cell cultures to an extent equivalent to FGF or medium conditioned by intact vitreous bodies (VBCM). Abolishing BMP2/4/7 signaling with noggin inhibited VBCM from upregulating fiber marker expression. Remarkably, noggin and anti-BMP antibodies also prevented purified FGF (but not unrelated stimuli) from upregulating the same fiber-specific proteins. This effect is attributable to inhibition of BMPs produced by the lens cells themselves. Although BMP signaling is required for FGF to enhance fiber differentiation, the converse is not true. Expression of noggin in the lenses of transgenic mice resulted in a postnatal block of epithelial-to-secondary fiber differentiation, with extension of the epithelial monolayer to the posterior pole of the organ. These results reveal the central importance of BMP in secondary fiber formation and show that although FGF may be necessary for this process, it is not sufficient. Differentiation of fiber cells, and thus proper vision, is dependent on cross-talk between the FGF and BMP signaling pathways.

Chlamydia Infection and Lymphomas: Association Beyond Ocular Adnexal Lymphomas Highlighted by Multiple Detection Methods

Chlamydia psittaci (Cp) has been associated to ocular adnexal lymphomas (OAL) with variable geographic distribution. Herein, we used multiple Chlamydia detection tools to identify Cp elementary bodies-containing cell and to assess Cp prevalence in both nodal and extranodal lymphomas. TETR-PCR, immunohistochemistry, immunofluorescence, electron microscopy, and laser-capture microdissection were done in 35 OALs to define their effect in Chlamydia detection and, moreover, to identify the Cp cellular carrier. Cp prevalence was screened by TETR-PCR in 205 extraorbital lymphomas and 135 nonneoplastic controls. Twenty-six (74%) OALs were associated with Cp infection: immunohistochemistry, immunofluorescence, and laser-capture microdissection-assisted PCR showed that monocytes/macrophages were the Cp carriers; electron microscopy showed the presence of intact Cp elementary bodies into these cells. Immunohistochemistry and TETR-PCR showed a 70% concordance rate (P = 0.001). Cp DNA was equally prevalent in non-OAL, nodal, and extranodal lymphomas: among the latter, it was more common in diffuse large B-cell lymphomas of the skin (P = 0.03) and Waldeyer's ring. This multiparametric approach shows, for the first time, that monocytes/macrophages are the carriers of Cp, Cp seems preferentially associated with lymphomas arising in organs primarily exposed to antigens. The clinical implications of these findings deserve to be prospectively investigated.

D1 Receptor Activation in the Mushroom Bodies Rescues Sleep-Loss-Induced Learning Impairments in Drosophila

Extended wakefulness disrupts acquisition of short-term memories in mammals. However, the underlying molecular mechanisms triggered by extended waking and restored by sleep are unknown. Moreover, the neuronal circuits that depend on sleep for optimal learning remain unidentified. Learning was evaluated with aversive phototaxic suppression. In this task, flies learn to avoid light that is paired with an aversive stimulus (quinine-humidity). We demonstrate extensive homology in sleep-deprivation-induced learning impairment between flies and humans. Both 6 hr and 12 hr of sleep deprivation are sufficient to impair learning in Canton-S (Cs) flies. Moreover, learning is impaired at the end of the normal waking day in direct correlation with time spent awake. Mechanistic studies indicate that this task requires intact mushroom bodies (MBs) and requires the dopamine D1-like receptor (dDA1). Importantly, sleep-deprivation-induced learning impairments could be rescued by targeted gene expression of the dDA1 receptor to the MBs. These data provide direct evidence that extended wakefulness disrupts learning in Drosophila. These results demonstrate that it is possible to prevent the effects of sleep deprivation by targeting a single neuronal structure and identify cellular and molecular targets adversely affected by extended waking in a genetically tractable model organism.

Inhibition of phago‐lysosome fusion by chlamydial elementary bodies is necessary for the early induction of type I IFNs in mouse macrophages

Two strains of Chlamydia, the mouse Chlamydia muridarum (MoPn) and the guinea pig Chlamydia caviae (GPIC) have equivalent growth rates in epithelial cells while only MoPn grows effectively in macrophages. To determine the early gene expression differences in mouse macrophages to these 2 organisms, micro array analysis was performed from RNA collected 3h post infection, followed by confirmation by real‐time quantitative RT‐PCR. Uninfected cells and cells treated with UV‐inactivated MoPn or GPIC were used as controls. Expression of pro‐inflammatory cytokines TNF‐α, IL‐6, IL1β, and inhibin β‐A expression were similar in GPIC or MoPn infected macrophages. However, only MoPn infection induced the expression of type I IFN and IFN response genes such as Egr‐1, IP‐10, and Gem GTPases. These responses were not observed in GPIC‐infected macrophages, or in UV‐inactivated MoPn and GPIC. To address the role of early bacterial and host interaction in the differences in gene expression, the infected macrophages were studied by electron microscopy (EM). EM analyses show the presence of intact chlamydial elementary bodies (EB) and reticulate bodies (RB) in endosomes of MoPn‐infected macrophages after 2h. In contrast, the GPIC EB's ended up in phagocytic vacuoles suggesting these EBs were killed. These data suggest that escape of MoPn chlamydial EB from lysosomal fusion is critical for the initiation of type I IFNs and IFN response genes. Further, these data suggest that induction of type I IFNs could be mediated by innate receptors interacting with the early endosomes containing chlamydiae.

Benzoxepine Esters as Precursors of the Wound‐Activated Chemical Defence of Mycena galopus

AbstractThe native precursors E/Z‐1a to E/Z‐1e and 2a–2e of the antifungally active benzoxepines E/Z‐1 and 2 have been isolated from intact fruiting bodies of Mycena galopus. These precursors turned out to be fatty acid esters of the benzoxepines (pterulones) E/Z‐1 and 2. Metabolic profiling of ethyl acetate extracts from intact fruiting bodies, from the latex and from mechanically wounded fruiting bodies revealed that the known antifungally active benzoxepines E/Z‐1 and the previously unknown benzoxepines 2 are only present in injured fruiting bodies. Intact fruiting bodies and the latex, however, exclusively contain the corresponding ester derivatives E/Z‐1a to E/Z‐1e and 2a–2e. Unlike the antifungally active benzoxepines E/Z‐1 and 2, the esters E/Z‐1a to E/Z‐1e present in intact fruiting bodies turned out to be inactive. Upon injury the inactive esters E/Z‐1a to E/Z‐1e are cleaved by a putative esterase, and the bioactive benzoxepines E/Z‐1 and 2 are released, suggesting a wound‐activated defence mechanism of M. galopus against yeasts and parasitic fungi. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)

Quality Control, Accuracy, and Prediction Capacity of Dual Energy X-ray Absorptiometry Variables and Data Acquisition

The acquisition of soft tissue measurements, fat (chemical) or adipose tissue (morphological) quantities, is essential in clinical research and nutritional status and its associated health risks. This has led to a proliferation of methods for the in-vivo determination of body composition. None of the indirect in-vivo approaches to estimate body adipose tissue has been validated against direct dissection data except for the skinfold.Since the development of DEXA as a measurement tool of bone density and mineral content for the detection of osteoporosis, it became a tool for measuring regional and whole body masses, lean tissue, and fat. A number of validation attempts have been made, however mostly against other indirect in-vivo techniques. The purpose of this study was to conduct an in-vitro validation and quality control of the data acquisition of DEXA using dissection as the criterion method. Fourteen porcine hind legs were scanned with DEXA, weighed in air and water and dissected into skin, adipose tissue, muscle and bone. Normal distribution, means, standard deviations, paired student t-test, Pearson correlation coefficients, interclass correlations test, and a Bland-Altman plot were used. The results show systematically good to excellent correlations between DEXA and dissection data acquisition (r(2)=0.75 to 0.99), but absolute indirect DEXA and direct dissection values are significantly different (p<0.05). DEXA overestimates total weight, lean mass, and fat free mass and underestimates both mass and % adipose tissue. DEXA provides erroneous values for bone density. Data produced by DEXA are morphological, not chemical values, as claimed by the manufacturer. This 'pilot' study indicates that a 'simple' combination of skin, adipose tissue, muscle, and bone from hind legs may give an incomplete picture of reality as one needs to measure viscera, connective tissue, and air pockets of the body. Validation studies with intact bodies are advised.

Red Pyrroloquinoline Alkaloids from the Mushroom Mycena haematopus

AbstractFour so far unknown red alkaloid pigments, haematopodin B (1) and the mycenarubins D (3), E (5) and F (6), were isolated from fruiting bodies of Mycena haematopus. The structures of these pyrroloquinoline alkaloids were established by 2D NMR spectroscopic and MS (ESI) methods. Their absolute configurations were determined by comparison of their CD spectra with that of haematopodin (2) or mycenarubin A (4). Metabolic profiling of the red pigments of intact and injured fruiting bodies revealed that the known degradation product haematopodin (2) originates from haematopodin B (1), which is the native main pigment of M. haematopus.(© Wiley‐VCH Verlag GmbH &amp; Co. KGaA, 69451 Weinheim, Germany, 2008)

Larval stenocephaly related to specialized feeding in the ant genera Amblyopone, Leptanilla and Myrmecina (Hymenoptera: Formicidae)

Larvae of the ant genera Amblyopone, Leptanilla and Myrmecina have unusually minute crania. This characteristic is here termed stenocephaly. To study it in detail larvae of the three genera were examined morphometrically and histologically in comparison with other non-stenocephalous larvae of the genera Cryptopone and Manica. The stenocephalous larvae are very specialized, in that their supra- and sub-esophageal ganglia are entirely displaced into the anterior thoracic segments, apparently as a consequence of the small cranial capacity. In Amblyopone and Leptanilla the small cranium appears to facilitate group feeding by a large number of larvae on the whole, intact bodies of the specialized centipede prey characteristic of these genera. In Myrmecina, the small, elongate head adapts the larva to consume the contents of the partly opened bodies of oribatid mites, which are the specialized prey of this genus. Therefore, stenocephaly seems to be a larval morphological adaptation facilitating specialized predation in these ants. These morphological specializations by ant larvae, and active larval involvement in prey dissection are possible because ants rear the larvae without confining them in cells. Acellular nests, a universal feature in Formicidae, may thus facilitate larval adaptations of benefit both to the larvae themselves and to the functionality of parental colonies, allowing them more efficiently to exploit accessible prey through well-integrated cooperation between adult and immature individuals.

The apneic threshold during non-REM sleep in dogs: sensitivity of carotid body vs. central chemoreceptors

The relative importance of peripheral vs. central chemoreceptors in causing apnea/unstable breathing during sleep is unresolved. This has never been tested in an unanesthetized preparation with intact carotid bodies. We studied three unanesthetized dogs during normal sleep in a preparation in which intact carotid body chemoreceptors could be reversibly isolated from the systemic circulation and perfused. Apneic thresholds and the CO(2) reserve (end-tidal Pco(2) eupneic - end-tidal Pco(2) apneic threshold) were determined using a pressure support ventilation technique. Dogs were studied when both central and peripheral chemoreceptors sensed transient hypocapnia induced by the pressure support ventilation and again with carotid body isolation such that only the central chemoreceptors sensed the hypocapnia. We observed that the CO(2) reserve was congruent with4.5 Torr when the carotid chemoreceptors sensed the transient hypocapnia but more than doubled (>9 Torr) when only the central chemoreceptors sensed hypocapnia. Furthermore, the expiratory time prolongations observed when only central chemoreceptors were exposed to hypocapnia differed from those obtained when both the central and peripheral chemoreceptors sensed the hypocapnia in that they 1) were substantially shorter for a given reduction in end-tidal Pco(2), 2) showed no stimulus: response relationship with increasing hypocapnia, and 3) often occurred at a time (>45 s) beyond the latency expected for the central chemoreceptors. These findings agree with those previously obtained using an identical pressure support ventilation protocol in carotid body-denervated sleeping dogs (Nakayama H, Smith CA, Rodman JR, Skatrud JB, Dempsey JA. J Appl Physiol 94: 155-164, 2003). We conclude that hypocapnia sensed at the carotid body chemoreceptor is required for the initiation of apnea following a transient ventilatory overshoot in non-rapid eye movement sleep.

Live‐attenuated influenza viruses as delivery vectors for Chlamydia vaccines

Effective delivery systems are needed to design efficacious vaccines against the obligate intracellular bacterial pathogen, Chlamydia trachomatis. Potentially effective delivery vehicles should promote the induction of adequate levels of mucosal T-cell and antibody responses that mediate long-term protective immunity. Antigen targeting to the nasal-associated lymphoid tissue (NALT) is effective for inducing high levels of specific immune effectors in the genital mucosa, and therefore suitable for vaccine delivery against genital chlamydial infection. We tested the hypothesis that live attenuated influenza A viruses are effective viral vectors for intranasal delivery of subunit vaccines against genital chlamydial infection. Recombinant influenza A/PR8/34 (H1N1) viruses were generated by insertion of immunodominant T-cell epitopes from chlamydial major outer membrane protein into the stalk region of the neuraminidase gene. Intranasal immunization of mice with viral recombinants resulted in a strong T helper 1 (Th1) response against intact chlamydial elementary bodies. Also, immunized mice enjoyed a significant state of protective immunity (P > 0.002) by shedding less chlamydiae and rapidly clearing the infection. Furthermore, a high frequency of Chlamydia-specific Th1 was measured in the genital mucosal and systemic draining lymphoid tissues within 24 hr after challenge of vaccinated mice. Moreover, multiple epitope delivery provided a vaccine advantage over single recombinants. Besides, long-term protective immunity correlated with the preservation of a robustly high frequency of specific Th1 cells and elevated immunoglobulin G2a in genital secretions. Because live attenuated influenza virus vaccines are safe and acceptable for human use, they may provide a new and reliable approach to deliver efficacious vaccines against sexually transmitted diseases.

Efficacy of spiracular infection of Helicoverpa armigera with its nucleopolyhedrovirus and its role in virus production

Baculoviruses are important microbial control agents of insects, with per os mode of infectivity. However, recently the spiracular infection of this virus group was suggested as an optimum method for virus production in grown up larvae. In this regard, a detailed evaluation of the spiracular infection with intact polyhedral inclusion bodies (PIB), alkali liberated virions and alkali liberated filtered virions of Helicoverpa armigera (Hubner) nucleopolyhedrovirus at 1 × 10 8, 1 × 10 7 and 2 × 10 6 PIB/ml concentrations was undertaken and compared with the standard diet surface treatment method. All the spiracle treatments resulted in larval death due to virus infection with alkali liberated virions causing higher mortality of larvae than alkali liberated filtered virions and intact PIB. Diet surface treatment method resulted in very high mortality as compared to spiracle treatment and among the different inoculum tested the intact PIB resulted in higher larval mortality. The PIB yield/larva in spiracle treatment was comparable with the diet surface treatment method, but due to very low larval mortality it resulted in low virus yield/100 inoculated larvae. Diet surface treatment with 5 × 10 7 PIB/ml concentration of virus resulted in the maximum yield of PIB/100 inoculated larvae. Low mortality, higher labour requirement and low amenability for mechanization for spiracle treatment method make it unviable for mass production of the virus in large scale compared to the standard diet surface treatment method.

A new component of attractive aggregation pheromone in the bean bug, Riptortus clavatus (Thunberg) (Heteroptera: Alydidae)

The attractive aggregation pheromone from males of the bean bug, Riptortus clavatus, has been identified as a blend of (E)-2-hexenyl (E)-2-hexenoate, (E)-2-hexenyl (Z)-3-hexenoate and tetradecyl isobutyrate (3 mix). When intact male bodies were extracted with hexane, octadecyl isobutyrate was detected in addition to the known 3 mix. Octadecyl isobutyrate was also detected in airborne volatiles using the glass-beaker collection method. In field experiments, the attractiveness of tetradecyl isobutyrate or 3 mix to R. clavatus was increased by the addition of octadecyl isobutyrate. These results suggest that octadecyl isobutyrate is one of the components of the attractive aggregation pheromone of R. clavatus, and may act as a synergistic composition in this bug.