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1.
Ecotoxicol Environ Saf ; 279: 116449, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38759532

RESUMO

Over the past few years, there has been growing interest in the ability of insect larvae to convert various organic side-streams containing mycotoxins into insect biomass that can be used as animal feed. Various studies have examined the effects of exposure to aflatoxin B1 (AFB1) on a variety of insect species, including the larvae of the black soldier fly (BSFL; Hermetia illucens L.; Diptera: Stratiomyidae) and the housefly (HFL; Musca domestica L.; Diptera: Muscidae). Most of these studies demonstrated that AFB1 degradation takes place, either enzymatic and/or non-enzymatic. The possible role of feed substrate microorganisms (MOs) in this process has thus far not been investigated. The main objective of this study was therefore to investigate whether biotransformation of AFB1 occurred and whether it is caused by insect-enzymes and/or by microbial enzymes of MOs in the feed substrate. In order to investigate this, sterile and non-sterile feed substrates were spiked with AFB1 and incubated either with or without insect larvae (BSFL or HFL). The AFB1 concentration was determined via LC-MS/MS analyses and recorded over time. Approximately 50% of the initially present AFB1 was recovered in the treatment involving BSFL, which was comparable to the treatment without BSFL (60%). Similar patterns were observed for HFL. The molar mass balance of AFB1 for the sterile feed substrates with BSFL and HFL was 73% and 78%, respectively. We could not establish whether non-enzymatic degradation of AFB1 in the feed substrates occurred. The results showed that both BSFL and substrate-specific MOs play a role in the biotransformation of AFB1 as well as in conversion of AFB1 into aflatoxin P1 and aflatoxicol, respectively. In contrast, HFL did not seem to contribute to AFB1 degradation. The obtained results contribute to our understanding of aflatoxin metabolism by different insect species. This information is crucial for assessing the safety of feeding fly larvae with feed substrates contaminated with AFB1 with the purpose of subsequent use as animal feed.


Assuntos
Aflatoxina B1 , Ração Animal , Biotransformação , Dípteros , Moscas Domésticas , Larva , Animais , Aflatoxina B1/metabolismo , Moscas Domésticas/metabolismo , Ração Animal/análise , Espectrometria de Massas em Tandem
2.
Mycotoxin Res ; 37(4): 279-295, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34618340

RESUMO

Novel protein sources for animal feed are needed, and the use of insects as feed ingredient is explored. The insect production sector offers opportunities for a circular and sustainable approach to feed production by upgrading waste or side streams into high-quality proteins. However, potential food or feed safety issues should be studied in advance. Mycotoxins, such as aflatoxin B1, are natural contaminants commonly found in agricultural crops and have proven to be detrimental to the agricultural industry, livestock, and human health. This systematic review aims to provide a comprehensive overview of the published evidence on effects of mycotoxin exposure on insect growth and survival, mycotoxin accumulation within the insect body, and metabolization of various mycotoxins by insects. The review includes 54 scientific articles published in the past 55 years, in total covering 32 insect species. The main findings are the following: (1) Insects of the order Coleoptera show lower mortality after exposure to aflatoxin B1 when compared to Lepidoptera and Diptera; (2) effects of mycotoxins on larval growth and survival are less detrimental in later larval stages; (3) accumulation of mycotoxins was low in most insect species; (4) mycotoxins are metabolized within the insect body, the degree of which depends on the particular mycotoxin and insect species; (5) cytochrome P450s are the main family of enzymes involved in biotransformation of mycotoxins in some insect species. Results of this review support an optimistic outlook for the use of mycotoxin-contaminated waste streams as substrate for insect rearing.


Assuntos
Micotoxinas , Ração Animal/análise , Animais , Biotransformação , Contaminação de Alimentos/análise , Humanos , Insetos/metabolismo , Larva/metabolismo , Micotoxinas/análise
3.
Trends Plant Sci ; 26(5): 496-508, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33358304

RESUMO

Light spectral composition influences plant growth and metabolism, and has important consequences for interactions with plant-feeding arthropods and their natural enemies. In greenhouse horticulture, light spectral composition can be precisely manipulated by light-emitting diodes (LEDs), and LEDs are already used to optimize crop production and quality. However, because light quality also modulates plant secondary metabolism and defense, it is important to understand the underlying mechanisms in the context of the growth-defense trade-off. We review the effects of the spectral composition of supplemental light currently used, or potentially used, in greenhouse horticulture on the mechanisms underlying plant growth and defense. This information is important for exploring opportunities to optimize crop performance and pest management, and thus for developing resilient crop-production systems.


Assuntos
Iluminação , Desenvolvimento Vegetal , Produção Agrícola , Luz , Plantas
4.
Phys Rev Lett ; 123(24): 244501, 2019 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-31922874

RESUMO

We report on the observation of gravity-capillary wave turbulence on the surface of a fluid in a high-gravity environment. By using a large-diameter centrifuge, the effective gravity acceleration is tuned up to 20 times Earth's gravity. The transition frequency between the gravity and capillary regimes is thus increased up to one decade as predicted theoretically. A frequency power-law wave spectrum is observed in each regime and is found to be independent of the gravity level and of the wave steepness. While the timescale separation required by weak turbulence is well verified experimentally regardless of the gravity level, the nonlinear and dissipation timescales are found to be independent of the scale, as a result of the finite size effects of the system (large-scale container modes) that are not taken currently into account theoretically.

5.
J Insect Physiol ; 95: 133-139, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27751886

RESUMO

Mating in the black soldier fly (BSF) is a visually mediated behaviour that under natural conditions occurs in full sunlight. Artificial light conditions promoting mating by BSF were designed based on the spectral characteristics of the compound eye retina. Electrophysiological measurements revealed that BSF ommatidia contained UV-, blue- and green-sensitive photoreceptor cells, allowing trichromatic vision. An illumination system for indoor breeding based on UV, blue and green LEDs was designed and its efficiency was compared with illumination by fluorescent tubes which have been successfully used to sustain a BSF colony for five years. Illumination by LEDs and the fluorescent tubes yielded equal numbers of egg clutches, however, the LED illumination resulted in significantly more larvae. The possibilities to optimize the current LED illumination system to better approximate the skylight illuminant and potentially optimize the larval yield are discussed.


Assuntos
Criação de Animais Domésticos/métodos , Olho Composto de Artrópodes/fisiologia , Dípteros/fisiologia , Iluminação , Células Fotorreceptoras de Invertebrados/fisiologia , Animais , Larva/fisiologia , Iluminação/normas , Reprodução , Gerenciamento de Resíduos
6.
Plant Biol (Stuttg) ; 18(6): 981-991, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27492059

RESUMO

Plants are part of biodiverse communities and frequently suffer from attack by multiple herbivorous insects. Plant responses to these herbivores are specific for insect feeding guilds: aphids and caterpillars induce different plant phenotypes. Moreover, plants respond differentially to single or dual herbivory, which may cascade into a chain of interactions in terms of resistance to other community members. Whether differential responses to single or dual herbivory have consequences for plant resistance to yet a third herbivore is unknown. We assessed the effects of single or dual herbivory by Brevicoryne brassicae aphids and/or Plutella xylostella caterpillars on resistance of plants from three natural populations of wild cabbage to feeding by caterpillars of Mamestra brassicae. We measured plant gene expression and phytohormone concentrations to illustrate mechanisms involved in induced responses. Performance of both B. brassicae and P. xylostella was reduced when feeding simultaneously with the other herbivore, compared to feeding alone. Gene expression and phytohormone concentrations in plants exposed to dual herbivory were different from those found in plants exposed to herbivory by either insect alone. Plants previously induced by both P. xylostella and B. brassicae negatively affected growth of the subsequently arriving M. brassicae. Furthermore, induced responses varied between wild cabbage populations. Feeding by multiple herbivores differentially activates plant defences, which has plant-mediated negative consequences for a subsequently arriving herbivore. Plant population-specific responses suggest that plant populations adapt to the specific communities of insect herbivores. Our study contributes to the understanding of plant defence plasticity in response to multiple insect attacks.


Assuntos
Afídeos/fisiologia , Brassica/fisiologia , Mariposas/fisiologia , Animais , Comportamento Alimentar , Herbivoria , Interações Hospedeiro-Parasita , Reguladores de Crescimento de Plantas/metabolismo
7.
Plant Biol (Stuttg) ; 17(2): 474-83, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25244063

RESUMO

Selected strains of non-pathogenic rhizobacteria can trigger induced systemic resistance (ISR) in plants against aboveground insect herbivores. However, the underlying mechanisms of plant-mediated interactions between rhizobacteria and herbivorous insects are still poorly understood. Using Arabidopsis thaliana Col-0-Pseudomonas fluorescens WCS417r as a model system, we investigated the performance and the molecular mechanisms underlying plant-mediated effects of rhizobacteria on the generalist caterpillar Mamestra brassicae and the specialist Pieris brassicae. Rhizobacteria colonisation of Arabidopsis roots resulted in decreased larval weight of M. brassicae, whereas no effect was observed on larval weight of P. brassicae. Using a jasmonic acid (JA)-impaired mutant (dde2-2), we confirmed the importance of JA in rhizobacteria-mediated ISR against M. brassicae. Interestingly, in some experiments we also observed rhizobacteria-induced systemic susceptibility to M. brassicae. The role of soil composition in the variable outcomes of microbe-plant-insect interactions was then assessed by comparing M. brassicae performance and gene transcription in plants grown in potting soil or a mixture of potting soil and sand in a 1:1 ratio. In a mixture of potting soil and sand, rhizobacteria treatment had a consistent negative effect on M. brassicae, whereas the effect was more variable in potting soil. Interestingly, at 24 h post-infestation (hpi) rhizobacteria treatment primed plants grown in a mixture of potting soil and sand for stronger expression of the JA- and ethylene-regulated genes PDF1.2 and HEL, respectively. Our study shows that soil composition can modulate rhizobacteria-plant-insect interactions, and is a factor that should be considered when studying these belowground-aboveground interactions.


Assuntos
Arabidopsis/fisiologia , Ciclopentanos/metabolismo , Herbivoria , Oxilipinas/metabolismo , Rhizobium/fisiologia , Microbiologia do Solo , Animais , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Arabidopsis/microbiologia , Proteínas de Arabidopsis/genética , Borboletas , Defensinas/genética , Regulação da Expressão Gênica de Plantas , Larva , Lepidópteros/fisiologia , Proteínas de Membrana/genética , Mutação , Raízes de Plantas/microbiologia , Pseudomonas fluorescens/fisiologia
8.
Med Vet Entomol ; 28(4): 407-13, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24797537

RESUMO

Mosquito repellents are used around the globe to protect against nuisance biting and disease-transmitting mosquitoes. Recently, there has been renewed interest in the development of repellents as tools to control the transmission of mosquito-borne diseases. We present a new bioassay for the accurate assessment of candidate repellent compounds, using a synthetic odour that mimics the odour blend released by human skin. Using DEET (N,N-diethyl-meta-toluamide) and PMD (p-menthane-3,8-diol) as reference compounds, nine candidate repellents were tested, of which five showed significant repellency to the malaria mosquito Anopheles gambiae sensu stricto (Diptera: Culicidae). These included: 2-nonanone; 6-methyl-5-hepten-2-one; linalool; δ-decalactone, and δ-undecalactone. The lactones were also tested on the yellow fever mosquito Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae), against which they showed similar degrees of repellency. We conclude that the lactones are highly promising repellents, particularly because these compounds are pleasant-smelling, natural products that are also present in human food sources.


Assuntos
Aedes/efeitos dos fármacos , Anopheles/efeitos dos fármacos , Repelentes de Insetos/farmacologia , Animais , Humanos , Odorantes
9.
Acta Biomater ; 9(5): 6653-62, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23415750

RESUMO

Introducing nanoroughness on various biomaterials has been shown to profoundly effect cell-material interactions. Similarly, physical forces act on a diverse array of cells and tissues. Particularly in bone, the tissue experiences compressive or tensile forces resulting in fluid shear stress. The current study aimed to develop an experimental setup for bone cell behavior, combining a nanometrically grooved substrate (200 nm wide, 50 nm deep) mimicking the collagen fibrils of the extracellular matrix, with mechanical stimulation by pulsatile fluid flow (PFF). MC3T3-E1 osteoblast-like cells were assessed for morphology, expression of genes involved in cell attachment and osteoblastogenesis and nitric oxide (NO) release. The results showed that both nanotexture and PFF did affect cellular morphology. Cells aligned on nanotexture substrate in a direction parallel to the groove orientation. PFF at a magnitude of 0.7 Pa was sufficient to induce alignment of cells on a smooth surface in a direction perpendicular to the applied flow. When environmental cues texture and flow were interacting, PFF of 1.4 Pa applied parallel to the nanogrooves initiated significant cellular realignment. PFF increased NO synthesis 15-fold in cells attached to both smooth and nanotextured substrates. Increased collagen and alkaline phosphatase mRNA expression was observed on the nanotextured substrate, but not on the smooth substrate. Furthermore, vinculin and bone sialoprotein were up-regulated after 1 h of PFF stimulation. In conclusion, the data show that interstitial fluid forces and structural cues mimicking extracellular matrix contribute to the final bone cell morphology and behavior, which might have potential application in tissue engineering.


Assuntos
Modelos Biológicos , Nanoestruturas , Osteoblastos/citologia , Células 3T3 , Animais , Sequência de Bases , Primers do DNA , Camundongos , Microscopia de Força Atômica , Reação em Cadeia da Polimerase Via Transcriptase Reversa
10.
Plant Biol (Stuttg) ; 14 Suppl 1: 83-90, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22348327

RESUMO

Beneficial microbes, such as plant growth-promoting rhizobacteria and mycorrhizal fungi, may have a plant-mediated effect on insects aboveground. The plant growth-promoting rhizobacterium Pseudomonas fluorescens can induce systemic resistance in Arabidopsis thaliana against several microbial pathogens and chewing insects. However, the plant-mediated effect of these beneficial microbes on phloem-feeding insects is not well understood. Using Arabidopsis as a model, we here report that P. fluorescens has a positive effect on the performance (weight gain and intrinsic rate of increase) of the generalist aphid Myzus persicae, while no effect was recorded on the crucifer specialist aphid Brevicoryne brassicae. Additionally, transcriptional analyses of selected marker genes revealed that in the plant-microbe interaction with M. persicae, rhizobacteria (i) prime the plant for enhanced expression of LOX2, a gene involved in the jasmonic acid (JA)-regulated defence pathway, and (ii) suppress the expression of ABA1, a gene involved in the abscisic acid (ABA) signalling pathway, at several time points. In contrast, almost no effect of the plant-microbe interaction with B. brassicae was found at the transcriptional level. This study presents the first data on rhizobacteria-induced systemic susceptibility to an herbivorous insect, supporting the pattern proposed for other belowground beneficial microbes and aboveground phloem feeders. Moreover, we provide further evidence that at the transcript level, soil-borne microbes modify plant-aphid interactions.


Assuntos
Afídeos/fisiologia , Arabidopsis/microbiologia , Arabidopsis/fisiologia , Pseudomonas fluorescens/fisiologia , Ácido Abscísico/metabolismo , Animais , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ciclopentanos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Herbivoria , Oxilipinas/metabolismo , Transdução de Sinais , Especificidade da Espécie
11.
Plant Biol (Stuttg) ; 13(2): 276-84, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21309974

RESUMO

When attacked by herbivores, plants release herbivore-induced plant volatiles (HIPV) that may function in direct defence by repelling herbivores or reducing their growth. Emission of HIPV may also contribute to indirect defence by attracting natural enemies of the herbivore. Here, cabbage (Brassica oleracea L.) plants (receiver plants) previously exposed to HIPV and subsequently induced through feeding by five Pieris brassicae L. caterpillars attracted more Cotesia glomerata L. parasitoids than control plants. HIPVs to which receiver plants had been exposed were emitted by B. oleracea infested with 50 P. brassicae caterpillars. Control plants had been exposed to volatiles from undamaged plants. In contrast, there were no differences in the attraction of wasps to receiver plants induced through feeding of one or ten larvae of P. brassicae compared to control plants. In addition, RT-PCR demonstrated higher levels of LIPOXYGENASE (BoLOX) transcripts in HIPV-exposed receiver plants. Exposure to HIPV from emitter plants significantly inhibited the growth rate of both P. brassicae and Mamestra brassicae caterpillars compared to growth rates of caterpillars feeding on control receiver plants. Our results demonstrate plant-plant signalling leading to priming of both indirect and direct defence in HIPV-exposed B. oleracea plants.


Assuntos
Brassica/metabolismo , Borboletas/crescimento & desenvolvimento , Lipoxigenase/genética , Proteínas de Plantas/genética , Compostos Orgânicos Voláteis/metabolismo , Animais , Brassica/efeitos dos fármacos , Brassica/genética , Ciclopentanos/metabolismo , Feminino , Regulação da Expressão Gênica de Plantas , Larva/crescimento & desenvolvimento , Oxilipinas/metabolismo , Pirazóis/farmacologia , Compostos Orgânicos Voláteis/farmacologia , Volatilização
12.
Med Vet Entomol ; 25(3): 247-55, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21108650

RESUMO

Host-seeking behaviour of the anthropophilic malaria vector Anopheles gambiae sensu stricto (Diptera: Culicidae) is mediated predominantly by olfactory cues. Several hundreds of odour components have been identified from human emanations, but only a few have been proven to act as attractants or synergists in the host-seeking behaviour of female An. gambiae. In previous work, aromatics, alcohols and ketones in human odours were found to elicit electrophysiological activity in antennal olfactory neurons of female An. gambiae. However, the behavioural effects of these compounds have not been investigated. In this study, behavioural responses of female An. gambiae to components of human breath, urine and sweat at a series of concentrations, or a single concentration in the case of acetone, were examined in combination with ammonia and L-lactic acid in a dual-choice olfactometer. The results showed that at specific concentrations 4-ethylphenol, indole, 3-methyl-1-butanol and two ketones inhibited the attractive effect of a mixture of ammonia and lactic acid. Acetone on its own was not attractive; however, when combined with lactic acid, the binary mixture was attractive. When combined with ammonia, acetone inhibited the attractiveness exerted by ammonia alone. Dodecanol and dimethyldisulphide did not affect the attraction exerted by ammonia and lactic acid at any of the concentrations tested. By contrast, a human-specific armpit odour, 7-octenoic acid, augmented the attraction exerted by the combination of ammonia and lactic acid at a specific dosage.


Assuntos
Anopheles/fisiologia , Comportamento Alimentar/efeitos dos fármacos , Feromônios/farmacologia , Suor/química , Urina/química , Adulto , Animais , Comportamento Alimentar/fisiologia , Feminino , Humanos , Odorantes , Feromônios/química
13.
J Insect Physiol ; 55(9): 788-92, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19414011

RESUMO

Behavioural and electrophysiological responsiveness to three chemically different secondary plant substances was studied in larvae of Pieris rapae L. (Lepidoptera: Pieridae). Three groups of caterpillars were studied that during their larval development were exposed to different rearing diets: an artificial diet or one of two host-plants, cabbage, Brassica oleracea, or nasturtium, Tropaeolum majus. In dual-choice leaf disc assays, caterpillars reared on cabbage were strongly deterred by the phenolic chlorogenic acid, the flavonol glycoside naringin and the alkaloid strychnine. However, behavioural plasticity was found in caterpillars reared on nasturtium or artificial diet in that these did not discriminate against chlorogenic acid. Caterpillars reared on the artificial diet were also significantly less sensitive to naringin and strychnine in the behavioural assay. Electrophysiological studies of the maxillary sensilla styloconica revealed that the deterrent neuron in the medial sensillum, but not in the lateral sensillum, of cabbage-reared caterpillars was more sensitive than the same neuron type of caterpillars reared on nasturtium or artificial diet. We conclude that (1) the diet-induced behavioural habituation to deterrents can at least partly be explained by chemosensory desensitisation of a generalist type of maxillary deterrent neuron; (2) behavioural cross-habituation to the three structurally diverse deterrent compounds can be traced back to cross-sensitivity for these compounds in the same gustatory neuron.


Assuntos
Brassica/química , Borboletas/fisiologia , Tropaeolum/química , Animais , Borboletas/crescimento & desenvolvimento , Comportamento Alimentar , Larva/crescimento & desenvolvimento , Larva/fisiologia
14.
J Microsc ; 233(2): 234-43, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19220689

RESUMO

We present a novel atomic force microscope (AFM) system, operational in liquid at variable gravity, dedicated to image cell shape changes of cells in vitro under hypergravity conditions. The hypergravity AFM is realized by mounting a stand-alone AFM into a large-diameter centrifuge. The balance between mechanical forces, both intra- and extracellular, determines both cell shape and integrity. Gravity seems to be an insignificant force at the level of a single cell, in contrast to the effect of gravity on a complete (multicellular) organism, where for instance bones and muscles are highly unloaded under near weightless (microgravity) conditions. However, past space flights and ground based cell biological studies, under both hypogravity and hypergravity conditions have shown changes in cell behaviour (signal transduction), cell architecture (cytoskeleton) and proliferation. Thus the role of direct or indirect gravity effects at the level of cells has remained unclear. Here we aim to address the role of gravity on cell shape. We concentrate on the validation of the novel AFM for use under hypergravity conditions. We find indications that a single cell exposed to 2 to 3 x g reduces some 30-50% in average height, as monitored with AFM. Indeed, in situ measurements of the effects of changing gravitational load on cell shape are well feasible by means of AFM in liquid. The combination provides a promising technique to measure, online, the temporal characteristics of the cellular mechano-response during exposure to inertial forces.


Assuntos
Forma Celular , Hipergravidade , Microscopia de Força Atômica/instrumentação , Microscopia de Força Atômica/métodos , Osteoblastos/citologia , Animais , Soluções Tampão , Células Cultivadas , Centrifugação , Meios de Cultura , Eletrônica , Camundongos , Osteoblastos/ultraestrutura , Software , Substâncias Viscoelásticas
15.
Cell Motil Cytoskeleton ; 65(2): 116-29, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17987657

RESUMO

This study evaluated in vitro the differences in morphological behaviour between fibroblast cultured on smooth and microgrooved substrata (groove depth: 0.5 mum, width: 1 mum), which were subjected to simulated microgravity. The aim of the study was to clarify which of these parameters was more dominant to determine cell behaviour. Morphological characteristics were investigated using scanning electron microscopy and fluorescence microscopy in order to obtain qualitative information on cell alignment. Expression of collagen type I, and alpha1-, beta1-, beta3-integrin were investigated by QPCR. Finally, immunoblotting was applied to visualise MAPK signalling pathways. Microscopy and image analysis showed that the fibroblasts aligned along the groove direction on all textured surfaces. On the smooth substrata, cells had spread out in a random fashion. The alignment of cells cultured on grooved surfaces under simulated microgravity, after 48 h of culturing appeared similar to those cultured at 1g, although cell shape was different. Analysis of variance proved that all main parameters: topography, gravity force, and time were significant. In addition, gene levels were reduced by simulated microgravity particularly those of beta3-integrin and collagen, however alpha-1 and beta-1 integrin levels were up-regulated. ERK1/2 was reduced in RPM, however, JNK/SAPK and p38 remained active. The members of the small GTPases family were stimulated under microgravity, particularly RhoA and Cdc42. The results are in agreement that application of microgravity to fibroblasts promotes a change in their morphological appearance and their expression of cell-substratum proteins through the MAPK intracellular signalling pathways.


Assuntos
Técnicas de Cultura de Células/métodos , Fibroblastos/enzimologia , Sistema de Sinalização das MAP Quinases/fisiologia , Ausência de Peso , Animais , Western Blotting , Células Cultivadas , Derme/citologia , Eletroforese em Gel de Poliacrilamida , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fibroblastos/ultraestrutura , Integrina alfa1/genética , Integrina alfa1/metabolismo , Integrina beta1/genética , Integrina beta1/metabolismo , Integrina beta3/genética , Integrina beta3/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Masculino , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas rho de Ligação ao GTP/metabolismo
16.
J Chem Ecol ; 33(10): 1831-44, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17828429

RESUMO

Interactions between butterflies and caterpillars in the genus Pieris and plants in the family Brassicaceae are among the best explored in the field of insect-plant biology. However, we report here for the first time that Pieris brassicae, commonly assumed to be a typical folivore, actually prefers to feed on flowers of three Brassica nigra genotypes rather than on their leaves. First- and second-instar caterpillars were observed to feed primarily on leaves, whereas late second and early third instars migrated via the small leaves of the flower branches to the flower buds and flowers. Once flower feeding began, no further leaf feeding was observed. We investigated growth rates of caterpillars having access exclusively to either leaves of flowering plants or flowers. In addition, we analyzed glucosinolate concentrations in leaves and flowers. Late-second- and early-third-instar P. brassicae caterpillars moved upward into the inflorescences of B. nigra and fed on buds and flowers until the end of the final (fifth) instar, after which they entered into the wandering stage, leaving the plant in search of a pupation site. Flower feeding sustained a significantly higher growth rate than leaf feeding. Flowers contained levels of glucosinolates up to five times higher than those of leaves. Five glucosinolates were identified: the aliphatic sinigrin, the aromatic phenylethylglucosinolate, and three indole glucosinolates: glucobrassicin, 4-methoxyglucobrassicin, and 4-hydroxyglucobrassicin. Tissue type and genotype were the most important factors affecting levels of identified glucosinolates. Sinigrin was by far the most abundant compound in all three genotypes. Sinigrin, 4-hydroxyglucobrassicin, and phenylethylglucosinolate were present at significantly higher levels in flowers than in leaves. In response to caterpillar feeding, sinigrin levels in both leaves and flowers were significantly higher than in undamaged plants, whereas 4-hydroxyglucobrassicin leaf levels were lower. Our results show that feeding on flower tissues, containing higher concentrations of glucosinolates, provides P. brassicae with a nutritional benefit in terms of higher growth rate. This preference appears to be in contrast to published negative effects of volatile glucosinolate breakdown products on the closely related Pieris rapae.


Assuntos
Borboletas/fisiologia , Comportamento Alimentar/fisiologia , Flores/química , Glucosinolatos/análise , Folhas de Planta/química , Animais , Peso Corporal , Genótipo , Larva/crescimento & desenvolvimento , Mostardeira/química , Mostardeira/genética , Fatores de Tempo
17.
Biomaterials ; 28(27): 3944-51, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17576010

RESUMO

The differences in morphological behaviour between fibroblasts cultured on smooth and nanogrooved substrata (groove depth: 5-350 nm, width: 20-1000 nm) have been evaluated in vitro. The aim of the study was to clarify to what extent cell guidance occurs on increasingly smaller topographies. Pattern templates were made using electron beam lithography, and were subsequently replicated in polystyrene cell culture material using solvent casting. The replicates were investigated with atomic force microscopy (AFM). After seeding with fibroblasts, morphological characteristics were investigated using scanning electron microscopy (SEM) and light microscopy, in order to obtain qualitative and quantitative information on cell alignment. AFM revealed that the nanogroove/ridge widths were replicated perfectly, although at deeper levels the grooves became more concave. The smooth substrata had no distinguishable pattern other than a roughness amplitude of 1 nm. Interestingly, microscopy and image analysis showed that fibroblast after 4 h had adjusted their shape according to nanotopographical features down to cut-off values of 100 nm width and 75 nm depth. After 24 h culturing time, fibroblasts would even align themselves on groove depths as shallow as 35 nm. It appears depth is the most essential parameter in cellular alignment on groove patterns with a pitch ratio of 1:1. On the smooth substrata, cells always spread out in a random fashion. Analysis of variance (ANOVA) demonstrated that both main parameters, topography and culturing time, were significant. We conclude that fibroblast cells cultured on nanotopography experience a threshold feature size of 35 nm, below this value contact guidance does no longer exist.


Assuntos
Materiais Biocompatíveis/química , Técnicas de Cultura de Células/métodos , Fibroblastos/citologia , Fibroblastos/fisiologia , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Engenharia Tecidual/métodos , Animais , Células Cultivadas , Masculino , Tamanho da Partícula , Ratos , Ratos Wistar , Propriedades de Superfície
18.
Cell Motil Cytoskeleton ; 64(3): 174-85, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17238131

RESUMO

This study evaluated in vitro the differences in morphological behaviour between fibroblast cultured on smooth and microgrooved substrata (groove depth: 0.5 microm, width: 1, 2, 5, and 10 microm), which were subjected to simulated microgravity. The aim of the study was to clarify which of these parameters was more dominant to determine cell behaviour. Morphological characteristics were investigated using scanning electron microscopy and fluorescence microscopy in order to obtain qualitative information on cell alignment and area. Confocal laser scanning microscopy visualised distribution of actin filaments and focal adhesion points. Finally, expression of collagen type I, fibronectin, and alpha1- and beta1-integrin were investigated by PCR. Microscopy and image analysis showed that the fibroblasts aligned along the groove direction on all textured surfaces. On the smooth substrata, cells had spread out in a random fashion. The alignment of cells cultured on grooved surfaces decreased under simulated microgravity, especially after 24 h of culturing. Cell surface area on grooved substrata were significantly smaller than on smooth substrata, but simulated microgravity on the grooved groups resulted in an enlargement of cell area. ANOVA was performed on all main parameters: topography, gravity force, and time. In this analysis, all parameters proved significant. In addition, gene levels were reduced by microgravity particularly those of beta1-integrin and fibronectin. From our data it is concluded that the fibroblasts primarily adjust their shape according to morphological environmental cues like substratum surface whilst a secondary, but significant, role is played by microgravity conditions.


Assuntos
Fibroblastos/citologia , Simulação de Ausência de Peso , Actinas/metabolismo , Animais , Polaridade Celular , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Fibroblastos/ultraestrutura , Regulação da Expressão Gênica , Masculino , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa
19.
J Gravit Physiol ; 14(1): P125-6, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18372731

RESUMO

A key requirement to enhance our understanding of the response of biological organisms to different levels of gravity is the availability of experimental systems that can simulate microgravity and hypergravity in ground-based laboratories. This paper compares the results obtained from analysing gene expression profiles of Drosophila in space versus those obtained in a random position machine (RPM) and by centrifugation. The correlation found validates the use of the RPM simulation technique to establish the effects of real microgravity on biological systems. This work is being extended to investigate Drosophila development in another gravity modifying instrument, the levitation magnet.


Assuntos
Adaptação Fisiológica/genética , Drosophila melanogaster/genética , Regulação da Expressão Gênica , Voo Espacial , Simulação de Ausência de Peso , Ausência de Peso , Animais , Centrifugação , Desenho de Equipamento , Perfilação da Expressão Gênica , Magnetismo , Reprodutibilidade dos Testes , Rotação , Simulação de Ausência de Peso/instrumentação , Simulação de Ausência de Peso/métodos
20.
Med Vet Entomol ; 20(3): 280-7, 2006 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17044878

RESUMO

Differences between human individuals in their attractiveness to female mosquitoes have been reported repeatedly, but the underlying mechanisms are not well understood. Skin emanations from 27 human individuals, collected on glass marbles, were tested against ammonia in a dual-choice olfactometer to establish their degrees of attractiveness to anthropophilic Anopheles gambiae s.s. Giles (Diptera: Culicidae) mosquitoes. Ammonia was used as a standard odour source because of its proven attractiveness to An. gambiae s.s. Skin emanations from most volunteers attracted significantly more mosquitoes than ammonia. There were clear differences in the attractiveness of skin emanations from different volunteers relative to that of ammonia, as well as in the strength of the trap entry response. Consistent differences were observed when emanations from the three most and the three least attractive volunteers were tested pairwise. No gender or age effect was found for relative attractiveness or trap entry response. Emanations from volunteers with higher behavioural attractiveness elicited higher electroantennogram response amplitudes in two pairs, but in a third pair a higher electroantennogram response was found for the less attractive volunteer. These results confirm that odour contributes to the differences in attractiveness of humans to mosquitoes.


Assuntos
Anopheles/fisiologia , Comportamento Animal/fisiologia , Odorantes , Animais , Feminino , Humanos , Masculino
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