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1.
Anim Biotelemetry ; 11(1): 8, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38800510

RESUMO

Background: The impact of biologging devices on the aerodynamics or hydrodynamics of animals is still poorly understood. This stands in marked contrast to the ever more extensive use of such technologies in wild-living animals. Recently, increasing concerns have been raised about the impairing effects of these devices on the animals concerned. In the early days of biotelemetry, attention was focused solely on reducing weight, but now aerodynamic effects are also increasingly being considered. To investigate these effects, we trained Northern Bald Ibises to fly in a wind tunnel in which we measured heart rate and dynamic body acceleration (VeDBA) as proxies for energy expenditure in relation to different logger shapes and wind flow directions. Results: Our data provide evidence that the position of biologging devices significantly influence the flight distances, and the shape of biologging devices has a considerable effect on heart rate and VeDBA, both of which have been used as proxies for energy expenditure. Unfavorable shape and positioning go beyond merely affecting the effort required during flapping flight. The energetically probably more important effect is that the devices impair the bird's ability to glide or soar and thus force them to perform the energetically much more demanding flapping flight more frequently. This effect was more pronounced in rising air than in horizontal airflow. A complementary study with wild Northern Bald Ibises during spring migration provides evidence that the position of the devices on the bird's back affects the length of the flight stages. Birds carrying the devices on the upper back, fixed by wing-loop harnesses, had significantly shorter flight stages compared to birds with a more caudally positioned device, fixed by leg-loop harnesses. Conclusion: The attachment of biologging devices on birds affects their performance and behavior and thus may influence their fitness and mortality. Our results show that detrimental effects can be reduced with relatively little effort, in particular through a strictly aerodynamic design of the housing and increased consideration of aerodynamics when attaching the device to the body. In birds, the attachment of biologging devices via leg loops to the lower back is clearly preferable to the common attachment via wing loops on the upper back, even if this affects the efficiency of the solar panels. Nevertheless, the importance of drag reduction may vary between systems, as the benefits of having a biologging devices close to the center of gravity may outweigh the increase in drag that this involves. Overall, more research is required in this field. This is both in the interest of animal welfare and of avoiding biasing the quality of the collected data. Supplementary Information: The online version contains supplementary material available at 10.1186/s40317-023-00322-5.

2.
J Ornithol ; 163(2): 599-610, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35464255

RESUMO

A blower-type wind tunnel for physiological bird flight experiments has been developed, constructed and evaluated. Since the birds to be investigated are rather big (Northern Bald Ibis, Geronticus eremita), the cross-sectional area of the test section measures 2.5 m × 1.5 m. The maximum achievable flow speed is approximately 16 ms-1. The wind tunnel exhibits a flexible outlet nozzle to provide up- and downdraft to allow for gliding and climbing flights. The current paper describes in detail the layout, design and construction of the wind tunnel including its control. Numerical simulations of the flow and measurements of the velocity distribution in the test section are presented. Apart from a non-homogeneous flow region in the mixing layer at the boundaries of the free jet, the test section exhibits a very even velocity distribution; the local speed deviates by less than two percent from the mean velocity. The turbulence intensity inside the test section was measured to be between 1 and 2%. As a constraint, a limited budget was available for the project. Four northern bald ibises were hand-raised and trained to fly in the wind tunnel. Supplementary Information: The online version contains supplementary material available at 10.1007/s10336-021-01945-2.

4.
J Neuroendocrinol ; 29(7)2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28514514

RESUMO

Djungarian hamsters are able to reduce their body weight by more than 30% in anticipation of the winter season. This particular adaptation to extreme environmental conditions is primarily driven by a natural reduction in day length and conserved under laboratory conditions. We used this animal model to investigate hypothalamic gene expression linked to body weight regulation behind this physiological phenomenon. After an initial collective short photoperiod (SP) adaptation for 14 weeks from a preceding long photoperiod (LP), hamsters were re-exposed to LP for either 6 or 14 weeks, followed by a second re-exposure to SP for 8 weeks. Our data showed that re-exposure to LP led to an increase in body weight. In the hypothalamus Dio2, Vimentin, Crbp1 and Grp50 expression increased, whereas expression of Dio3, Mct8 and Srif decreased. The changes in body weight and gene expression were reversible in most hamsters after a further re-exposure to SP following 6 or 14 weeks in LP. Interestingly, after 14 weeks in LP, body weight loss was pronounced in six hamsters re-exposed to SP, but five hamsters did not respond. In nonresponding hamsters, a different gene expression pattern was manifested, with the exception of Dio2, which was reduced not only in SP re-exposed hamsters, but also in hamsters maintained in LP. Taken together, these data suggest that body weight regulation appears to be tightly linked to a co-ordinated regulation of several genes in the hypothalamus, including those involved in thyroid hormone metabolism.


Assuntos
Peso Corporal/fisiologia , Regulação da Expressão Gênica , Hipotálamo/metabolismo , Phodopus/fisiologia , Fotoperíodo , Estações do Ano , Animais , Cricetinae , Feminino , Iodeto Peroxidase/genética , Iodeto Peroxidase/metabolismo , Masculino , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Vimentina/genética , Vimentina/metabolismo , Iodotironina Desiodinase Tipo II
5.
J Neuroendocrinol ; 28(11)2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27755687

RESUMO

Endothermic mammals and birds require intensive energy turnover to sustain high body temperatures and metabolic rates. To cope with the energetic bottlenecks associated with the change of seasons, and to minimise energy expenditure, complex mechanisms and strategies are used, such as daily torpor and hibernation. During torpor, metabolic depression and low body temperatures save energy. However, these bouts of torpor, lasting for hours to weeks, are interrupted by active 'euthermic' phases with high body temperatures. These dynamic transitions require precise communication between the brain and peripheral tissues to defend rheostasis in energetics, body mass and body temperature. The hypothalamus appears to be the major control centre in the brain, coordinating energy metabolism and body temperature. The sympathetic nervous system controls body temperature by adjustments of shivering and nonshivering thermogenesis, with the latter being primarily executed by brown adipose tissue. Over the last decade, comparative physiologists have put forward integrative studies on the ecophysiology, biochemistry and molecular regulation of energy balance in response to seasonal challenges, food availability and ambient temperature. Mammals coping with such environments comprise excellent model organisms for studying the dynamic regulation of energy metabolism. Beyond the understanding of how animals survive in nature, these studies also uncover general mechanisms of mammalian energy homeostasis. This research will benefit efforts of translational medicine aiming to combat emerging human metabolic disorders. The present review focuses on recent advances in the understanding of energy balance and its neuronal and endocrine control during the most extreme metabolic fluctuations in nature: daily torpor and hibernation.


Assuntos
Metabolismo Energético , Hibernação , Homeostase , Mamíferos/fisiologia , Torpor , Animais , Encéfalo/fisiologia , Ritmo Circadiano , Sistema Endócrino/fisiologia , Humanos , Estações do Ano
6.
ACS Appl Mater Interfaces ; 7(46): 25955-60, 2015 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-26536909

RESUMO

We report on the attainment of broadband white light emission from a host-guest light-emitting electrochemical cell, comprising a blue-emitting conjugated polymer as the majority host and a red-emitting small-molecule triplet emitter as the minority guest. An analysis of the energy structure reveals that host-to-guest energy transfer can be effectuated by both Förster and Dexter processes, and through a careful optimization of the active material composition partial energy transfer and white emission is accomplished at a low guest concentration of 0.5%. By adding a small amount of a yellow-emitting conjugated polymer to the active material, white light emission with a high color rendering index of 79, and an efficiency of 4.3 cd/A at significant luminance (>200 cd/m(2)), is realized.

7.
J Am Chem Soc ; 135(9): 3647-52, 2013 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-23398145

RESUMO

We report a novel and generic approach for attaining white light from a single-emitter light-emitting electrochemical cell (LEC). With an active-layer comprising a multifluorophoric conjugated copolymer (MCP) and an electrolyte designed to inhibit MCP energy-transfer interactions during LEC operation, we are able to demonstrate LECs that emit broad-band white light with a color rendering index of 82, a correlated-color temperature of 4000 K, and a current conversion efficacy of 3.8 cd/A. It is notable that this single-emitter LEC configuration eliminates color-drift problems stemming from phase separation, which are commonly observed in conventional blended multiemitter devices. Moreover, the key role of the electrolyte in limiting undesired energy-transfer reactions is highlighted by the observation that an electrolyte-free organic light-emitting diode comprising the same MCP emits red light.


Assuntos
Técnicas Eletroquímicas , Luz , Técnicas Eletroquímicas/instrumentação , Eletrólitos/química , Estrutura Molecular , Polímeros/química , Espectrometria de Fluorescência , Temperatura
8.
J Neuroendocrinol ; 25(2): 190-7, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22967033

RESUMO

The production of bioactive peptides from biologically inactive precursors involves extensive post-translational processing, including enzymatic cleavage by proteolytic peptidases. Endoproteolytic prohormone-convertases initially cleave the precursors of many neuropeptides at specific amino acid sequences to generate intermediates with basic amino acid extensions on their C-termini. Subsequently, the related exopeptidases, carboxypeptidases D and E (CPD and CPE), are responsible for removing these amino acids before the peptides achieve biological activity. We investigated the effect of photoperiod on the processing of the neuropeptide precursor pro-opiomelanocortin (POMC) and its derived neuropeptides, α-melanocyte-stimulating hormone (MSH) and ß-endorphin (END), within the hypothalamus of the seasonal Siberian hamster (Phodopus sungorus). We thus compared hypothalamic distribution of CPD, CPE, α-MSH and ß-END using immunohistochemistry and measured the enzyme activity of CPE and concentrations of C-terminally cleaved α-MSH in short-day (SD; 8 : 16 h light/dark) and long-day (LD; 16 : 8 h light/dark) acclimatised hamsters. Increased immunoreactivity (-IR) of CPE, as well as higher CPE activity, was observed in SD. This increase was accompanied by more ß-END-IR cells and substantially higher levels of C- terminally cleaved α-MSH, as determined by radioimmunoassay. Our results suggest that exoproteolytic cleavage of POMC-derived neuropeptides is tightly regulated by photoperiod in the Siberian hamster. Higher levels of biological active α-MSH- and ß-END in SD are consistent with the hypothesis that post-translational processing is a key event in the regulation of seasonal energy balance.


Assuntos
Carboxipeptidase H/metabolismo , Neuropeptídeos/metabolismo , Phodopus/fisiologia , Fotoperíodo , Animais , Núcleo Arqueado do Hipotálamo/metabolismo , Peso Corporal/fisiologia , Cricetinae , Masculino , Phodopus/metabolismo , Pró-Opiomelanocortina/metabolismo , Processamento de Proteína Pós-Traducional , Estações do Ano , Especificidade por Substrato , alfa-MSH/metabolismo , beta-Endorfina/metabolismo
9.
J Neuroendocrinol ; 24(7): 991-8, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22487258

RESUMO

Siberian hamsters are seasonal mammals that survive a winter climate by making adaptations in physiology and behaviour. This includes gonadal atrophy, reduced food intake and body weight. The underlying central mechanisms responsible for the physiological adaptations are not fully established but involve reducing hypothalamic tri-iodthyronine (T3) levels. Juvenile Siberian hamsters born or raised in short days (SD) respond in a similar manner, although with an inhibition of gonadal development and growth instead of reversing an established long day (LD) phenotype. Using juvenile male hamsters, the present study aimed to investigate whether the central mechanisms are similar before the establishment of the mature LD phenotype. By in situ hybridisation, we examined the response of genes involved in thyroid hormone (Dio2 and Dio3, which determine hypothalamic T3 levels) and glucose/glutamate metabolism in the ependymal layer, histamine H3 receptor and VGF as representatives of the highly responsive dorsomedial posterior arcuate nucleus (dmpARC), and somatostatin, a hypothalamic neuropeptide involved in regulating the growth axis. Differential gene expression of type 2 and type 3 deiodinase in the ependymal layer, histamine H3 receptor in the dmpARC and somatostatin in the ARC was established by the eighth day in SD. These changes are followed by alterations in glucose metabolism related genes in the ependymal layer by day 16 and increased secretogranin expression in the dmpARC by day 32. In conclusion, our data demonstrate similar but rapid and highly responsive changes in gene expression in the brain of juvenile Siberian hamsters in response to a switch from LD to SD. The data also provide a temporal definition of gene expression changes relative to physiological adaptations of body weight and testicular development and highlight the likely importance of thyroid hormone availability as an early event in the adaptation of physiology to a winter climate in juvenile Siberian hamsters.


Assuntos
Regulação da Expressão Gênica , Hipotálamo/metabolismo , Phodopus/genética , Fotoperíodo , Fatores Etários , Animais , Animais Lactentes , Núcleo Arqueado do Hipotálamo/metabolismo , Peso Corporal/fisiologia , Cricetinae , Masculino , Tamanho do Órgão , Phodopus/metabolismo , Phodopus/fisiologia , Estações do Ano , Testículo/anatomia & histologia , Fatores de Tempo , Desmame
10.
Virology ; 403(1): 56-66, 2010 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-20444481

RESUMO

Ebola virus (EBOV) causes severe hemorrhagic fevers in humans and non-human primates. While the role of the EBOV major matrix protein VP40 in morphogenesis is well understood, nothing is known about its contributions to the regulation of viral genome replication and/or transcription. Similarly, while it was reported that the minor matrix protein VP24 impairs viral genome replication, it remains unclear whether it also regulates transcription, since all common experimental systems measure the combined products of replication and transcription. We have developed systems that allow the independent monitoring of viral transcription and replication, based on qRT-PCR and a replication-deficient minigenome. Using these systems we show that VP24 regulates not only viral genome replication, but also transcription. Further, we show for the first time that VP40 is also involved in regulating these processes. These functions are conserved among EBOV species and, in the case of VP40, independent of its budding or RNA-binding functions.


Assuntos
Ebolavirus/fisiologia , Regulação Viral da Expressão Gênica , Nucleoproteínas/fisiologia , RNA Viral/biossíntese , Transcrição Gênica , Proteínas do Core Viral/fisiologia , Proteínas Virais/fisiologia , Animais , Linhagem Celular , Chlorocebus aethiops , Perfilação da Expressão Gênica , Genes Reporter , Humanos , Luciferases/genética , Luciferases/metabolismo
11.
Am J Physiol Regul Integr Comp Physiol ; 296(3): R631-9, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19144754

RESUMO

In the adult brain, leptin regulates energy homeostasis primarily via hypothalamic circuitry that affects food intake and energy expenditure. Evidence from rodent models has demonstrated that during early postnatal life, leptin is relatively ineffective in modulating these pathways, despite the high circulating levels and the presence of leptin receptors within the central nervous system. Furthermore, in recent years, a neurotrophic role for leptin in the establishment of energy balance circuits has emerged. The precise way in which leptin exerts these effects, and the site of leptin action, is unclear. To provide a detailed description of the development of energy balance systems in the postnatal rat in relation to leptin concentrations during this time, endogenous leptin levels were measured, along with gene expression of leptin receptors and energy balance neuropeptides in the medial basal hypothalamus, using in situ hybridization. Expression of leptin receptors and both orexigenic and anorexigenic neuropeptides increased in the arcuate nucleus during the early postnatal period. At postnatal day 4 (P4), we detected dense leptin receptor expression in ependymal cells of the third ventricle (3V), which showed a dramatic reduction over the first postnatal weeks, coinciding with marked morphological changes in this region. An acute leptin challenge robustly induced suppressor of cytokine signaling-3 expression in the 3V of P4 but not P14 animals, revealing a clear change in the location of leptin action over this period. These findings suggest that the neurotrophic actions of leptin may involve signaling at the 3V during a restricted period of postnatal development.


Assuntos
Animais Recém-Nascidos/fisiologia , Metabolismo Energético/fisiologia , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/metabolismo , Leptina/metabolismo , Neuropeptídeos/metabolismo , Receptores para Leptina/metabolismo , Animais , Núcleo Arqueado do Hipotálamo/crescimento & desenvolvimento , Núcleo Arqueado do Hipotálamo/metabolismo , Glicemia/metabolismo , Ensaio de Imunoadsorção Enzimática , Epêndima/citologia , Epêndima/metabolismo , Feminino , Hibridização In Situ , Insulina/sangue , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Wistar , Proteína 3 Supressora da Sinalização de Citocinas , Proteínas Supressoras da Sinalização de Citocina/biossíntese , Proteínas Supressoras da Sinalização de Citocina/genética , Terceiro Ventrículo/citologia , Terceiro Ventrículo/crescimento & desenvolvimento , Terceiro Ventrículo/metabolismo
12.
Vaccine ; 26(7): 956-65, 2008 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-18164519

RESUMO

The most effective countermeasure against a pandemic originating from a highly pathogenic avian influenza virus (HPAIV) is immunoprophylaxis of the human population. We present here a new approach for the development of a pandemic HPAIV live vaccine. Using reverse genetics, we replaced the polybasic hemagglutinin cleavage site of an H7N7 HPAIV with an elastase motif. This mutant was strictly elastase-dependent, grew equally well as the wild-type in cell culture and was attenuated in mice unlike the lethal wild-type. Immunization at 10(6)pfu dosage protected mice against disease and induced sterile immunity; vaccination with homosubtypic or heterosubtypic reassortants led to cross-protection. These observations demonstrate that a mutated hemagglutinin requiring elastase cleavage can serve as an attenuating component of a live vaccine against HPAIV.


Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Vírus da Influenza A Subtipo H7N7/genética , Vacinas contra Influenza , Mutação , Elastase Pancreática/genética , Vacinas Atenuadas , Animais , Anticorpos Antivirais/análise , Anticorpos Antivirais/sangue , Linhagem Celular , Chlorocebus aethiops , Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Humanos , Imunidade nas Mucosas , Vírus da Influenza A Subtipo H7N7/enzimologia , Vírus da Influenza A Subtipo H7N7/patogenicidade , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Vacinas contra Influenza/imunologia , Camundongos , Dados de Sequência Molecular , Testes de Neutralização , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Elastase Pancreática/metabolismo , Recombinação Genética , Análise de Sequência de DNA , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/genética , Vacinas Atenuadas/imunologia , Células Vero
13.
J Neuroendocrinol ; 19(12): 1001-7, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18001330

RESUMO

Seasonal animals use different strategies to reduce energy expenditure in the face of reduced seasonal food availability. For example, the ground squirrel enters a hibernation state with reduced metabolism, hypothermia and suppressed central nervous system activity, whereas the Djungarian hamster (Phodopus sungorus) employs daily bouts of torpor associated with reduced body temperature and energy expenditure. Studies in the hibernating ground squirrel implicate an increase in histamine synthesis and histamine H(3) receptor expression in the brain as a central mechanism governing hibernation. In the present study, we demonstrate an up-regulation of H(3) receptors in several brain nuclei in the Djungarian hamster during bouts of daily torpor, a shallow form of hypothermia, suggesting that histaminergic pathways may play a general role in maintaining low body temperature and torpor state in mammals. These regions include the arcuate nucleus, dorsomedial hypothalamus, suprachiasmatic nucleus, dorsal lateral geniculate nucleus and tuberomammillary nucleus. Interestingly, expression of the mRNA for orexins, a group of neuropeptides that increase wakefulness, remains unchanged during the arousal from daily torpor, suggesting that this classic 'arousal' pathway is not involved in the transition from a hypothermic to the euthermic state.


Assuntos
Hibernação/fisiologia , Neuropeptídeos/biossíntese , Receptores Histamínicos H3/biossíntese , Animais , Nível de Alerta/fisiologia , Temperatura Corporal/fisiologia , Cricetinae , Hibridização In Situ , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Orexinas , Phodopus , Fotoperíodo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptores Histamínicos H3/genética
14.
Cell Mol Life Sci ; 62(16): 1863-70, 2005 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16003493

RESUMO

Muscle satellite cells are believed to form a stable, self-renewing pool of stem cells in adult muscle where they function in tissue growth and repair. A regulatory disruption of growth and differentiation of these cells is assumed to result in tumor formation. Here we provide for the first time evidence that sonic hedgehog (Shh) regulates the cell fate of adult muscle satellite cells in mammals. Shh promotes cell division of satellite cells (and of the related model C2C12 cells) and prevents their differentiation into multinucleated myotubes. In addition, Shh inhibits caspase-3 activation and apoptosis induced by serum deprivation. These effects of Shh are reversed by simultaneous administration of cyclopamine, a specific inhibitor of the Shh pathway. Taken together, Shh acts as a proliferation and survival factor of satellite cells in the adult muscle. Our results support the hypothesis of the rhabdomyosarcoma origin from satellite cells and suggest a role for Shh in this process.


Assuntos
Células Satélites de Músculo Esquelético/citologia , Transativadores/metabolismo , Animais , Apoptose , Caspase 3 , Inibidores de Caspase , Caspases/metabolismo , Diferenciação Celular , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Proteínas Hedgehog , Camundongos , Camundongos Transgênicos , Fibras Musculares Esqueléticas/citologia , Fibras Musculares Esqueléticas/fisiologia , Células Satélites de Músculo Esquelético/fisiologia , Transdução de Sinais , Transativadores/antagonistas & inibidores , Transativadores/genética , Alcaloides de Veratrum/farmacologia
15.
SAR QSAR Environ Res ; 13(1): 89-110, 2002 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12074394

RESUMO

The choice of an appropriate structure coding scheme is the secret to success in QSAR studies. Depending on the problem at hand, 2D or 3D descriptors have to be chosen; the consideration of electronic effects might be crucial, conformational flexibility has to be of special concern. Artificial neural networks, both with unsupervised and with supervised learning schemes, are powerful tools for establishing relationships between structure and physical, chemical, or biological properties. The EROS system for the simulation of chemical reactions is briefly presented and its application to the degradation of s-triazine herbicides is shown. It is further shown how the simulation of chemical reactions can be combined with the simulation of infrared spectra for the efficient identification of the structure of degradation products.


Assuntos
Técnicas de Apoio para a Decisão , Modelos Químicos , Previsões , Herbicidas/efeitos adversos , Herbicidas/farmacologia , Raios Infravermelhos , Conformação Molecular , Relação Estrutura-Atividade , Triazinas
16.
J Virol ; 74(14): 6316-23, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10864641

RESUMO

The hemagglutinin (HA) of fowl plague virus A/FPV/Rostock/34 (H7N1) carries two N-linked oligosaccharides attached to Asn123 and Asn149 in close vicinity to the receptor-binding pocket. In previous studies in which HA mutants lacking either one (mutants G1 and G2) or both (mutant G1,2) glycosylation sites had been expressed from a simian virus 40 vector, we showed that these glycans regulate receptor binding affinity (M. Ohuchi, R. Ohuchi, A. Feldmann, and H. D. Klenk, J. Virol. 71:8377-8384, 1997). We have now investigated the effect of these mutations on virus growth using recombinant viruses generated by an RNA polymerase I-based reverse genetics system. Two reassortants of influenza virus strain A/WSN/33 were used as helper viruses to obtain two series of HA mutant viruses differing only in the neuraminidase (NA). Studies using N1 NA viruses revealed that loss of the oligosaccharide from Asn149 (mutant G2) or loss of both oligosaccharides (mutant G1,2) has a pronounced effect on virus growth in MDCK cells. Growth of virus lacking both oligosaccharides from infected cells was retarded, and virus yields in the medium were decreased about 20-fold. Likewise, there was a reduction in plaque size that was distinct with G1,2 and less pronounced with G2. These effects could be attributed to a highly impaired release of mutant progeny viruses from host cells. In contrast, with recombinant viruses containing N2 NA, these restrictions were much less apparent. N1 recombinants showed lower neuraminidase activity than N2 recombinants, indicating that N2 NA is able to partly overrule the high-affinity binding of mutant HA to the receptor. These results demonstrate that N-glycans flanking the receptor-binding site of the HA molecule are potent regulators of influenza virus growth, with the glycan at Asn149 being dominant and that at Asn123 being less effective. In addition, we show here that HA and NA activities need to be highly balanced in order to allow productive influenza virus infection.


Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Vírus da Influenza A/crescimento & desenvolvimento , Neuraminidase/genética , Animais , Bovinos , Linhagem Celular , Galinhas , Eritrócitos/virologia , Glicosilação , Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Técnicas In Vitro , Vírus da Influenza A/genética , Vírus da Influenza A/metabolismo , Mutagênese Sítio-Dirigida , Neuraminidase/metabolismo , Receptores Virais/genética , Receptores Virais/metabolismo , Recombinação Genética , Ensaio de Placa Viral
17.
J Chem Inf Comput Sci ; 40(2): 482-94, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10761155

RESUMO

Organic reactions can be run under a variety of conditions, from laboratory experiments, through technical processes, to combinatorial chemistry. The scope is further extended when the metabolism of compounds and the reactions in the mass spectrometer are included. We present here several concepts: reactors, phases, and modes, which, together with a kinetic modeling, allow the treatment of such a broad scope of organic reactions. These concepts have been implemented in a knowledge-based system, EROS. Several applications of this system to the wide world of organic reactions are given.

18.
J Virol ; 72(5): 3554-9, 1998 May.
Artigo em Inglês | MEDLINE | ID: mdl-9557635

RESUMO

The hemagglutinin (HA) of fowl plague virus was lengthened and shortened by site-specific mutagenesis at the cytoplasmic tail, and the effects of these modifications on HA functions were analyzed after expression from a simian virus 40 vector. Elongation of the tail by the addition of one to six histidine (His) residues did not interfere with intracellular transport, glycosylation, proteolytic cleavage, acylation, cell surface expression, and hemadsorption. However, the ability to induce syncytia at a low pH decreased dramatically depending on the number of His residues added. Partial fusion (hemifusion), assayed by fluorescence transfer from octadecylrhodamine-labeled erythrocyte membranes, was also reduced, but even with the mutant carrying six His residues, significant transfer was observed. However, when the formation of fusion pores was examined with hydrophilic fluorescent calcein, transfer from erythrocytes to HA-expressing cells was not observed with the mutant carrying six histidine residues. The addition of different amino acids to the cytoplasmic tail of HA caused an inhibitory effect similar to that caused by the addition of His. On the other hand, a mutant lacking the cytoplasmic tail was still able to fuse at a reduced level. These results demonstrate that elongation of the cytoplasmic tail interferes with the formation and enlargement of fusion pores. Thus, the length of the cytoplasmic tail plays a critical role in the fusion process.


Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Histidina/metabolismo , Vírus da Influenza A/metabolismo , Fusão de Membrana/fisiologia , Aminoácidos , Animais , Transporte Biológico , Linhagem Celular , Membrana Celular/metabolismo , Chlorocebus aethiops , Citoplasma/metabolismo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/química , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Histidina/genética , Vírus da Influenza A/genética , Líquido Intracelular , Mutagênese
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