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Larval Cryptothelea gloverii (Lepidoptera: Psychidae), an arthropod predator and herbivore on Florida citrus.

The orange bagworm (OBW), Cryptothelea gloverii (Packard) (Lepidoptera: Psychidae) was previously reported feeding on citrus fruit and foliage and preying upon the camphor scale Pseudaonidia duplex (Cockerell) (Homoptera: Coccidae). In this study using laboratory assays, OBW preyed upon citrus rust mite, Phyllocoptruta oleivora (Ashmead) (Acari: Eriophyidae) and consumed eggs and adults of both P. oleivora and Panonychus citri (McGregor) (Acari: Tetranychidae), two important pest mites on Florida citrus. OBW was also observed to feed on the purple scale, Lepidosaphes beckii (Newman) (Homoptera: Diaspididae) and on a fungus (Penicillium sp.). OBW fed on orange and grapefruit leaves by starting from the border and eating part of the leaf, by chewing holes, or consuming the outer epithelium of either the axial or abaxial surface of the leaf without penetrating through the leaf. OBW was observed in orange orchards in association with fruit extensively russeted by P. oleivora feeding. Laboratory assays revealed that OBW larvae preferred to feed on oranges infested with P. oleivora rather than on clean fruits that were free of mite feeding damage. Feeding damage to citrus fruit by OBW larvae results in one to several holes being eaten into the rind or albedo, without damage to the fruit sacs.

Animals↗

Changes in the composition of phospholipid fatty acids and sterols of maize root in response to monoterpenes.

Terpenes are thought to be important in plant plant interactions because of their phytotoxic action on seed germination and growth. Herein, the effects of five volatile monoterpenes on root sterols and phospholipid fatty acid (PLFA) composition have been studied during maize seedling germination. The investigated monoterpenes (camphor, 1-8 cineole, geraniol, menthol, and thymol) were applied at their respective IC80 (concentration causing 80% inhibition). They quantitatively affected free sterols and PLFA composition, thus producing an increase in the percentage of unsaturated PLFAs, stigmasterol of the free sterol fraction, and saturated steryl ester fatty acids. Alcoholic and nonalcoholic monoterpenes appeared to have different modes of action. The former affected unsaturated fatty acid and stigmasterol to a greater extent, and accordingly they could interfere in seedling growth by changes in the membrane lipids.

Cell Membrane↗

Antimicrobial activities of the essential oils of various plants against tomato late blight disease agent Phytophthora infestans.

The aim of this study was to find an alternative to synthetic fungicides currently used in the control of devastating oomycete pathogen Phytophthora infestans, causal agent of late blight disease of tomato. Antifungal activities of essential oils obtained from aerial parts of aromatic plants such as oregano (Origanum syriacum var. bevanii), thyme (Thymbra spicata subsp. spicata), lavender (Lavandula stoechas subsp. stoechas), rosemary (Rosmarinus officinalis), fennel (Foeniculum vulgare), and laurel (Laurus nobilis), were investigated against P. infestans. Both contact and volatile phase effects of different concentrations of the essential oils used were determined by using two in vitro methods. Chemical compositions of the essential oils were also determined by GC-MS analysis. Major compounds found in essential oils of thyme, oregano, rosemary, lavender, fennel and laurel were carvacrol (37.9%), carvacrol (79.8), borneol (20.4%), camphor (20.2%), anethole (82.8%) and 1,8-cineole (35.5%), respectively. All essential oils were found to inhibit the growth of P. infestans in a dose-dependent manner. Volatile phase effect of oregano and thyme oils at 0.3 microg/ml air was found to completely inhibit the growth of P. infestans. Complete growth inhibition of pathogen by essential oil of fennel, rosemary, lavender and laurel was, however, observed at 0.4-2.0 microg/ml air concentrations. For the determination of the contact phase effects of the tested essential oils, oregano, thyme and fennel oils at 6.4 microg/ml were found to inhibit the growth of P. infestans completely. Essential oils of rosemary, lavender and laurel were inhibitory at relatively higher concentrations (12.8, 25.6, 51.2 microg/ml respectively). Volatile phase effects of essential oils were consistently found to be more effective on fungal growth than contact phase effect. Sporangial production was also inhibited by the essential oil tested. Light and scanning electron microscopic (SEM) observation on pathogen hyphae, exposed to both volatile and contact phase of oil, revealed considerable morphological alterations in hyphae such as cytoplasmic coagulation, vacuolations, hyphal shrivelling and protoplast leakage.

Gas Chromatography-Mass Spectrometry↗

Oxime-metabolizing activity of liver aldehyde oxidase.

Liver aldehyde oxidase in the presence of its electron donor exhibited a significant oxime-metabolizing activity toward some different types of oximes under anaerobic conditions. Acetophenone oxime and salicylaldoxime were exclusively converted to the corresponding oxo compounds, whereas benzamidoxime was converted to the corresponding ketimine. With d-camphor oxime, the formation of both the corresponding oxo compound and ketimine was observed. Stoichiometric studies showed that the formation of oxo compounds is accompanied by nearly equimolar ammonia. We propose a mechanism of oxime biotransformation that liver aldehyde oxidase catalyzes the reduction of oximes to the corresponding ketimines which in turn undergo, depending on their chemical stability, nonenzymatic hydrolysis to the corresponding oxo compounds and ammonia.

Aldehyde Oxidase↗

The reduction of putidaredoxin reductase by reduced pyridine nucleotides.

Putidaredoxin reductase (PdR), an FAD-containing protein, mediates the transfer of electrons from NADH to putidaredoxin in the cytochrome P-450cam-dependent oxidation of camphor. Using stopped-flow spectrophotometry, reduction of putidaredoxin reductase by NADH (70 microM) at 4 degrees C appeared to be a pseudo-first-order process with a rate constant in excess of 600 s-1. The reduction of putidaredoxin reductase by NADPH was much slower with a second-order rate constant of 530 s-1 M-1 at 4 degrees C. The reduction of the enzyme was monitored at several wavelengths: 455 nm to follow flavin reduction; 700 nm to follow the appearance of the long-wavelength charge-transfer complex; and 513 nm to detect the presence of a semiquinone form of the flavoprotein. There was no apparent semiquinone formation observed during reduction. The charge-transfer complex can be formed in the presence of NAD+, whereas, no charge-transfer band could be detected when PdR was reduced with NADPH. The titration of chemically or NADPH-reduced putidaredoxin reductase with either a stoichiometric or an excess amount of NAD+ resulted in the formation of a charge-transfer complex, indicating that the reduced form of PdR has a high affinity for NAD+ regardless of the method of reduction. The data presented indicate that putidaredoxin reductase is reduced without the formation of semiquinone intermediate and, upon reduction, forms a tight complex with NAD+. The Keq for the reduction of PdR by NADPH is 1.1 and the midpoint potential for this reaction is -317 +/- 5 mV.

Electrochemistry↗

Molecular mechanisms of odor-sensing. II. Studies of fractions from olfactory tissue scrapings capable of sensitizing artificial lipid membranes to action of odorants.

From the preparation obtained by ultrasonic disintegration of frog olfactory epithelium scraping, a fraction capable of sensitizing artificial phospholipid membranes to the action of some chemicals assumed to be odorants in frogs was isolated. In the presence of the active fraction, the membranes respond to the addition of camphor, linalool and musk ambrette by an increased permeability to Na+ and Ca2+. The main component of the active fraction is a nucleo-protein with molecular weight of no less than 100 000.

Animals↗

Receptor cell responses to odorants: similarities and differences among odorants.

An extensive exploration of the discriminating properties of olfactory receptors cells has been conducted in our laboratory through the statistical processing of single cell responses recorded in the frog's olfactory epithelium. Similarities between odorant stimulating properties could be demonstrated by comparing the complex response profiles of receptor cells, resulting in the concept of odorant groups which depicts consistent relationships found between some odorous compounds. The recording technique limits the number of odorants which can be tested and compared within the same series of experiments. Thus, based upon previous studies we have chosen a specific set of 20 odorants whose characteristics are known but, up to this time, have not been compared in the same experimental system. These odorants were: acetophenone, anisole, n-butanol, DL-camphor, cyclodecanone, 1,8-cineole, p-cymene, D-citronellol, n-heptanol, isoamyl acetate, isovaleric acid, D-limonene, methyl amylketone, L-menthol, phenol, thiophenol, pyridine, thymol, cyclohexanol, cyclohexanone. The pattern of similarities between these odorants, as delineated with the aid of correlation coefficient computation, factor analysis and non-hierarchical taxonomy, confirms the reality of several odorant groups previously suggested and describes their cross-relationships. The receptor mechanisms underlying these odorant groups are discussed. Because reliable prominent features of the olfactory stimulus space can be established on an objective basis, the findings are proposed as a reference for future studies on other aspects of olfactory discrimination.

Animals↗

Identification of specific pathways of communication between the CNS and NK cell system.

The specific signals and pathways utilized by the natural killer (NK) cell system and the central nervous system (CNS) that results in the conditioned response (CR) is not clearly understood. Single trial conditioning of the NK cell activity provides us with a model to probe the mechanisms of communication between two major systems (Immune and CNS) which are involved in the health and disease of the individual. The studies show that the IFN-beta molecules possess the properties attributed to the unconditioned stimulus (US). IFN-beta can penetrate the CNS and evoke the elevation of NK cell activity in the spleen. This unconditioned response (UR) can be linked to a specific conditioned stimulus (CS). Specific odors such as camphor provide a neural pathway for the CS to associate with the US. Evidence is presented that in conditioning there are two locations where memory develops. The CS/US association is made centrally and its memory is stored at a central location, but the memory for the specificity of the odor is presumably stored in the olfactory bulbs. The CS recalls the CR by triggering the olfactory neural pathway which, in turn, signals the hypothalamic-pituitary axis to release mediators that modulate the activity of NK cells in the spleen. These results imply that through conditioning one has direct input into the regulatory hypothalamus that controls the internal environment of the organism and the health and disease of the individual. Consequently, it is not inconceivable that through this approach we might be able to alter the course of a disease process.

Animals↗

Diffusion of medicaments within root canal dentin. A scanning electron microscopic study.

Root canals of 21 maxillary human teeth were enlarged with reamers and flushed alternatively with sodium hypochlorite and hydrogen peroxide. After its mechanical preparation, root canals were dressed with camphorated paramonochloropherol and/or formocresol delivered by paper points. All three medicaments crystallize both on dentin walls and inside dentinal tubules. The clinical implications of these findings are discussed.

Adult↗

High-performance liquid chromatographic method for the separation and quantitative estimation of anti-parasitic melaminophenyl arsenical compounds.

A sensitive high-performance liquid chromatographic method has been developed for the separation and quantitative estimation of melaminophenyl arsenical drugs used in the treatment of human and veterinary trypanosomiases. Five clinically relevant compounds (melarsoprol, melarsonyl potassium, cymelarsan, melarsen oxide, and sodium melarsen) can be separated with an octadecylsilane reverse-phase column and detected down to 10 pmol per injection. The chromatographic system utilizes a propanol gradient in water, with lithium camphor sulphonate as ion modifier, and detection by ultraviolet absorbance at 280 nm. Preliminary results indicate that the arsenical compounds can be extracted and concentrated from serum using octadecylsilane solid-phase extraction cartridges. This method represents the first specific and sensitive assay system for separating and quantitatively estimating this important class of antiparasitic agents.

Arsenicals↗

Indomethacin and sodium carbonate effects on conditioned fever and NK cell activity.

The augmentation of natural killer (NK) cell activity and elevation of body temperature (fever) can both be conditioned using camphor odor as the conditioned stimulus (CS) and poly I:C as the unconditioned stimulus (US). While both responses can be conditioned in parallel fashion as shown previously, our results indicate the conditioned learning of these responses may not follow along a common path. We found that injection of a 1% solution of sodium carbonate was able to consistently block the CS/US learning of the NK cell response but did not block conditioning of the fever response. In contrary fashion, mice treated with indomethacin (which inhibits prostaglandin-induced fever) dissolved in the sodium carbonate solution did not learn in consistent fashion the fever response. However, indomethacin-treated animals were able to recall the NK cell response. These results support the view that although the same mediator, IFN-beta, is responsible for the conditioned learning of the NK cell and fever responses both the learning and recall of the responses are initiated along separate pathways.

Animals↗

Conditioning of the allogeneic cytotoxic lymphocyte response.

Allogeneic cytotoxic T-lymphocyte (CTL) response can be obtained following immunization of BALB/c mice with C57BL/6 spleen cells. We investigated the possibility of behaviorally conditioning this response by associating the C57BL/6 spleen cell immunization [unconditioned stimulus (US)] with camphor odor [conditioned stimulus (CS)]. We reported the possible mechanisms involved in the conditioning of natural killer cell activity. Similar approaches were used to investigate the mechanisms that participate in the conditioned CTL activity. The first mechanism of investigation utilized opioid receptor blockers naltrexone and quaternary naltrexone. Naltrexone, which blocks both the central and peripheral opioid receptors, blocked the recall of the conditioned response, whereas quaternary naltrexone, which does not penetrate the blood-brain barrier, was unable to block the conditioned response, demonstrating that centrally located opioid receptors play a role in the recall of the conditioned response. The studies are of interest because they indicate that resistance or susceptibility to various diseases such as cancer, autoimmunity, and infectious diseases might be influenced by the regulatory network of the CNS.

Animals↗

Reliability criteria of a bioassay using rats trained to detect estrus-specific odor in cow urine.

An odor discrimination apparatus was used to quantify the reaction of rats against odor differences of estrous and diestrous urine of cows. Rats were trained to release impulses from a microswitch when they came into contact with estrous but not diestrous urine. After training, the discrimination ability was highly significant (P< or = 0.001) between both urine types. The reaction was specific for cow urine, and no significant discrimination occurred for estrous and diestrous urine from pigs or goats, or that of women. The discrimination of urine diluted with water was still highly significant up to 1:25. Another group of rats was trained to detect camphor. Decreasing concentrations were detected down to 0.1/microg/ml when diluted in water, and down to 1/microg/ml when diluted in estrous urine. Once trained to discriminate between estrous and diestrous urine, rats maintained this ability for 12 mo. Thus, a bioassay was set up to accompany fractionation steps in the laboratory which could lead to the isolation of estrous pheromones in cow urine.

Journal Article↗

Cytogenetic assays for mitotic poisons. The grasshopper embryo system for volatile liquids.

The present study demonstrates the usefulness of grasshopper (Melanoplus sanguinipes) embryos for detecting mitotic arrest and anaphase abnormalities induced by volatile liquids. Eight compounds were tested with this system. Embryos in vivo (intact chorions) were suspended in sealed chambers and the agents, in various amounts, were placed on the bottom of the chambers. The agents, after evaporation, filled the ambient air in which the embryos respired. After a period of exposure, individual embryos were fixed for squash preparations to determine the mitotic index (MI) and the ratio of anaphases to metaphases (A/M). If an agent completely arrested mitosis at metaphase, the A/M ratio would be zero and the MI would be elevated if the agent does not prevent interphase cells from entering mitosis. Two common organic solvents, benzene and toluene, and two common anesthetics, halothane and chloroform, were found to be potent mitotic arrestants in the present system. Colchicine-like mitoses (c-mitoses) were observed in embryos after treatment of the above four agents; all of these agents except toluene were found to induce an accumulation of c-mitoses. Amyl acetate, 95% ethanol, deionized water, and camphor (in 10% ethanol) failed to show mitosis-arresting effects on the embryos. Abnormal anaphases with lagging chromosomes and multipolar spindles were observed in embryos which showed partial metaphase block after exposure to concentrations of benzene lower than that required for a complete block. Benzene, toluene, halothane, and chloroform have been implicated as teratogens and carcinogens, but are negative as mutagens, clastogens, and SCE inducers. Thus, the carcinogenic activity of these compounds appears to result from their action on cell division mechanisms.

Animals↗

Role of arcuate nucleus of the hypothalamus in the acquisition of association memory between the CS and US.

A single trial association protocol was used to demonstrate a conditioned increase in natural killer (NK) cell activity. The signals used were odor of camphor as the conditioned stimulus (CS) and polyinosinic-polycytidylic acid (poly I:C) as the unconditioned stimulus (US). This model has been used to dissect the underlying mechanisms of interaction between the central nervous system (CNS) and the immune system (IS) and vice versa. Here, we demonstrate the potential role played by the arcuate nucleus of the hypothalamus in the acquisition of association memory between the CS and the US. Chemical destruction of the arcuate nucleus with monosodium glutamate (MSG) was used for this purpose. Mice with arcuate nucleus lesion prior to the association protocol did not demonstrate a conditioned increase in NK cell activity. However, the lesion has no effect if produced prior to exposure to the CS at recall. These studies demonstrate the significant role played by the hypothalamus (arcuate nucleus) in a conditioned response.

Adrenocorticotropic Hormone↗

Arecoline a muscarinic cholinergic agent conditions central pathways that modulate natural killer cell activity.

The central nervous system plays an active role in the regulation of the immune system. Modulation of immune activities appears to be in part under the control of the hypothalamic-pituitary-adrenal (HPA) axis. We investigated the effect of a muscarinic cholinergic agonist, arecoline, which stimulates the secretion of corticotropin-releasing hormone (CRF) and adrenocorticotropic hormone (ACTH) on the immune system. In this report we demonstrate that peripherally administered arecoline or ACTH can increase activity of pre-activated NK cells. Second, we show that central administration of arecoline at a dose too low to alter peripheral events is sufficient to induce a significant increase in the activity of pre-activated natural killer (NK) cells. Finally, we demonstrate by using a Pavlovian conditioning paradigm that the pairing of a novel odor (camphor) with administration of arecoline can be used to alter NK cell activity. Subsequent to the conditioning trial, exposure to the odor alone is sufficient to raise NK cell activity. From these observations, we infer that the pathway(s) that are conditioned reside in sites located within the CNS and the conditioned response is evoked in the peripheral compartment (NK cell activity).

Adrenocorticotropic Hormone↗

Lack of mutagenicity to S. typhimurium of neopentyl bromide and pentaerythrityl tetrachloride: relation to chemical structure.

Neopentyl bromide and pentaerythrityl tetrachloride were shown here to be non-mutagenic to 7 strains of Salmonella typhimurium. These inactivities are reflected in the inability of either compound to produce a colour reaction in the chemical alkylation test of Epstein (4-nitrobenzylpyridine, NBP). It is suggested that the lack of biological reactivity of these two alkyl halides is due to steric crowding of the halogen group. These findings are significant within the context of the potent mutagenicity of mono-haloalkanes in general. The similarity between the odour of pentaerythrityl tetrachloride and camphor is discussed.

Chemical Phenomena↗

Olfactory receptor responses of the nymphal American cockroach to sex pheromones and their mimics.

1. EAG responses to highly purified sex pheromones (periplanone-A and -B), sex pheromone mimics [germacrene-D, (+)-verbanyl acetate and (+)-trans-verbenyl acetate] and general odor (camphor) were recorded from both sexes of adult and three nymphal stages (7, 10 and 11th instars) of the American cockroach. 2. The M/F ratios were evaluated for each stage by stimulation with the above chemicals. 3. The ratio values indicated undeveloped sex pheromone receptor on the antennae of 7th male instar and females of all the stages. On the other hand, precursory development of the receptor was expected on the antennae of males of the old-aged nymphal instars and full development on the adult male antennae.

Animals↗