RESUMO
A single-hydrophone ocean glider was deployed within a cabled hydrophone array to demonstrate a framework for estimating population density of fin whales (Balaenoptera physalus) from a passive acoustic glider. The array was used to estimate tracks of acoustically active whales. These tracks became detection trials to model the detection function for glider-recorded 360-s windows containing fin whale 20-Hz pulses using a generalized additive model. Detection probability was dependent on both horizontal distance and low-frequency glider flow noise. At the median 40-Hz spectral level of 97 dB re 1 µPa2/Hz, detection probability was near one at horizontal distance zero with an effective detection radius of 17.1 km [coefficient of variation (CV) = 0.13]. Using estimates of acoustic availability and acoustically active group size from tagged and tracked fin whales, respectively, density of fin whales was estimated as 1.8 whales per 1000 km2 (CV = 0.55). A plot sampling density estimate for the same area and time, estimated from array data alone, was 1.3 whales per 1000 km2 (CV = 0.51). While the presented density estimates are from a small demonstration experiment and should be used with caution, the framework presented here advances our understanding of the potential use of gliders for cetacean density estimation.
Assuntos
Baleia Comum , Animais , Cetáceos , Probabilidade , Acústica , Aeronaves , Vocalização AnimalRESUMO
Many animals increase the intensity of their vocalizations in increased noise. This response is known as the Lombard effect. While some previous studies about cetaceans report a 1 dB increase in the source level (SL) for every dB increase in the background noise level (NL), more recent data have not supported this compensation ability. The purpose of this study was to calculate the SLs of humpback whale song units recorded off Hawaii and test for a relationship between these SLs and background NLs. Opportunistic recordings during 2012-2017 were used to detect and track 524 humpback whale encounters comprised of 83 974 units on the U.S. Navy's Pacific Missile Range Facility hydrophones. Received levels were added to their estimated transmission losses to calculate SLs. Humpback whale song units had a median SL of 173 dB re 1 µPa at 1 m, and SLs increased by 0.53 dB/1 dB increase in background NLs. These changes occurred in real time on hourly and daily time scales. Increases in ambient noise could reduce male humpback whale communication space in the important breeding area off Hawaii. Since these vocalization changes may be dependent on location or behavioral state, more work is needed at other locations and with other species.
Assuntos
Jubarte , Acústica , Animais , Havaí , Masculino , Oceanos e Mares , Vocalização AnimalRESUMO
Minke whales were acoustically detected, localized, and tracked on the U.S. Navy's Pacific Missile Range Facility from 2012 to 2017. Animal source levels (SLs) were estimated by adding transmission loss estimates to measured received levels of 42 159 individual minke whale boings. Minke whales off Hawaii exhibited the Lombard effect in that they increased their boing call intensity in increased background noise. Minke whales also decreased the variance of the boing call SL in higher background noise levels. Although the whales partially compensated for increasing background noise, they were unable or unwilling to increase their SLs by the same amount as the background noise. As oceans become louder, this reduction in communication space could negatively impact the health of minke whale populations. The findings in this study also have important implications for acoustic animal density studies, which may use SL to estimate probability of detection.
RESUMO
Acoustically equipped deep-water mobile autonomous platforms can be used to survey for marine mammals over intermediate spatiotemporal scales. Direct comparisons to fixed recorders are necessary to evaluate these tools as passive acoustic monitoring platforms. One glider and two drifting deep-water floats were simultaneously deployed within a deep-water cabled hydrophone array to quantitatively assess their survey capabilities. The glider was able to follow a pre-defined track while float movement was somewhat unpredictable. Fin whale (Balaenoptera physalus) 20 Hz pulses were recorded by all hydrophones throughout the two-week deployment. Calls were identified using a template detector, which performed similarly across recorder types. The glider data contained up to 78% fewer detections per hour due to increased low-frequency flow noise present during glider descents. The glider performed comparably to the floats and fixed recorders at coarser temporal scales; hourly and daily presence of detections did not vary by recorder type. Flow noise was related to glider speed through water and dive state. Glider speeds through water of 25 cm/s or less are suggested to minimize flow noise and the importance of glider ballasting, detector characterization, and normalization by effort when interpreting glider-collected data and applying it to marine mammal density estimation are discussed.
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Time difference of arrival methods for acoustically localizing multiple marine mammals have been applied to recorded data from the Navy's Pacific Missile Range Facility in order to localize and track calls attributed to Bryde's whales. Data were recorded during the months of August-October 2014, and 17 individual tracks were identified. Call characteristics were compared to other Bryde's whale vocalizations from the Pacific Ocean, and locations of the recorded signals were compared to published visual sightings of Bryde's whales in the Hawaiian archipelago. Track kinematic information, such as swim speeds, bearing information, track duration, and directivity, was recorded for the species. The intercall interval was also established for most of the tracks, providing cue rate information for this species that may be useful for future acoustic density estimate calculations.
RESUMO
The biosonar signals of two free-swimming Atlantic bottlenose dolphins performing a complex sonar search for a bottom target in San Diego Bay were compared with the biosonar signals of a dolphin performing a target discrimination task in a net pen in the same bay. A bite-plate device carried by the free-swimming dolphins supported a hydrophone that extended directly in front of the dolphin. A biosonar measuring tool attached to the bite plate measured the outgoing biosonar signals while the dolphins conducted sonar searches. Each of the free-swimming dolphins used different biosonar search strategy in solving the problem and the dolphins' biosonar signals reflect the difference in strategy. The dolphin in the pen stationed in a hoop while echolocating on a target 6 m away and reported if the indentation on a spherical target was directed toward it. The signals were parameterized by determining the peak-to-peak source levels, source energy flux density, peak frequency, center frequency, root-mean-square (rms) bandwidth, rms duration, and the Q of the signals. Some parameters were similar for the free-swimming and stationary dolphins while some were significantly different, suggesting biosonar signals used by free-swimming animals may be different than signals used by dolphins in a pen.
Assuntos
Golfinho Nariz-de-Garrafa/fisiologia , Golfinho Nariz-de-Garrafa/psicologia , Ecolocação , Natação , Vocalização Animal , Acústica/instrumentação , Animais , Discriminação Psicológica , Masculino , Processamento de Sinais Assistido por Computador , Espectrografia do Som , TransdutoresRESUMO
Time difference of arrival (TDOA) methods for acoustically localizing multiple marine mammals have been applied to recorded data from the Navy's Pacific Missile Range Facility in order to localize and track humpback whales. Modifications to established methods were necessary in order to simultaneously track multiple animals on the range faster than real-time and in a fully automated way, while minimizing the number of incorrect localizations. The resulting algorithms were run with no human intervention at computational speeds faster than the data recording speed on over forty days of acoustic recordings from the range, spanning multiple years. Spatial localizations based on correlating sequences of units originating from within the range produce estimates having a standard deviation typically 10 m or less (due primarily to TDOA measurement errors), and a bias of 20 m or less (due primarily to sound speed mismatch). An automated method for associating units to individual whales is presented, enabling automated humpback song analyses to be performed.
Assuntos
Algoritmos , Jubarte/fisiologia , Processamento de Sinais Assistido por Computador/instrumentação , Vocalização Animal/fisiologia , Animais , Automação , Simulação por Computador , Sinais (Psicologia) , Comportamento Alimentar/fisiologia , Modelos Teóricos , Método de Monte Carlo , Oceano Pacífico , Densidade Demográfica , Localização de Som , Espectrografia do Som/instrumentação , Espectrografia do Som/métodos , Especificidade da Espécie , TransdutoresRESUMO
Minke whales (Balaenoptera acutorostrata) were acoustically detected and localized via their boing calls using 766 h of recorded data from 24 hydrophones at the U.S. Navy's Pacific Missile Range Facility located off Kauai, Hawaii. Data were collected before, during, and after naval undersea warfare training events, which occurred in February over three consecutive years (2011-2013). Data collection in the during periods were further categorized as phase A and phase B with the latter being the only period with naval surface ship activities (e.g., frigate and destroyer maneuvers including the use of mid-frequency active sonar). Minimum minke whale densities were estimated for all data periods based upon the numbers of whales acoustically localized within the 3780 km(2) study area. The 2011 minimum densities in the study area were: 3.64 whales [confidence interval (CI) 3.31-4.01] before the training activity, 2.81 whales (CI 2.31-3.42) for phase A, 0.69 whales (CI 0.27-1.8) for phase B and 4.44 whales (CI 4.04-4.88) after. The minimum densities for the phase B periods were highly statistically significantly lower (p < 0.001) from all other periods within each year, suggesting a clear response to the phase B training. The phase A period results were mixed when compared to other non-training periods.
Assuntos
Acústica , Ecolocação , Monitoramento Ambiental/métodos , Militares , Baleia Anã/fisiologia , Ruído dos Transportes/efeitos adversos , Navios , Vocalização Animal , Animais , Ecolocação/classificação , Baleia Anã/classificação , Baleia Anã/psicologia , Movimento (Física) , Densidade Demográfica , Processamento de Sinais Assistido por Computador , Som , Espectrografia do Som , Especificidade da Espécie , Fatores de Tempo , Vocalização Animal/classificaçãoRESUMO
Immunological memory is characterized by heightened immunoglobulin (Ig) G antibody production caused in part by enhanced plasma cell formation conferred by conserved transmembrane and cytoplasmic segments in isotype-switched IgG B cell receptors. We tested the hypothesis that the IgG tail enhances intracellular B cell antigen receptor (BCR) signaling responses to antigen by analyzing B cells from Ig transgenic mice with IgM receptors or chimeric IgMG receptors containing the IgG tail segment. The IgG tail segment enhanced intracellular calcium responses but not tyrosine or extracellular signal-related kinase (ERK) phosphorylation. Biochemical analysis and crosses to CD22-deficient mice established that IgG tail enhancement of calcium and antibody responses, as well as marginal zone B cell formation, was not due to diminished CD22 phosphorylation or inhibitory function. Microarray profiling showed no evidence for enhanced signaling by the IgG tail for calcium/calcineurin, ERK, or nuclear factor kappaB response genes and little evidence for any enhanced gene induction. Instead, almost half of the antigen-induced gene response in IgM B cells was diminished 50-90% by the IgG tail segment. These findings suggest a novel "less-is-more" hypothesis to explain how switching to IgG enhances B cell memory responses, whereby decreased BCR signaling to genes that oppose marginal zone and plasma cell differentiation enhances the formation of these key cell types.
Assuntos
Sequência Conservada , Memória Imunológica/imunologia , Receptores de Antígenos de Linfócitos B/química , Receptores de Antígenos de Linfócitos B/metabolismo , Receptores de IgG/química , Receptores de IgG/metabolismo , Transdução de Sinais , Animais , Linfócitos B/citologia , Linfócitos B/imunologia , Linfócitos B/metabolismo , Cálcio/metabolismo , Diferenciação Celular , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Imunoglobulina M/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fosfotirosina/metabolismo , Receptores de Antígenos de Linfócitos B/genética , Receptores de Antígenos de Linfócitos B/imunologia , Receptores de IgG/genética , Receptores de IgG/imunologia , Lectina 2 Semelhante a Ig de Ligação ao Ácido Siálico/genética , Lectina 2 Semelhante a Ig de Ligação ao Ácido Siálico/metabolismo , Transdução de Sinais/imunologia , Ativação TranscricionalRESUMO
Humoral immune responses are thought to be enhanced by complement-mediated recruitment of the CD21-CD19-CD81 coreceptor complex into the B cell antigen receptor (BCR) complex, which lowers the threshold of B cell activation and increases the survival and proliferative capacity of responding B cells. To investigate the role of the CD21-CD35 complement receptors in the generation of B cell memory, we analyzed the response against viral particles derived from the bacteriophage Qbeta in mice deficient in CD21-CD35 (Cr2(-/-)). Despite highly efficient induction of early antibody responses and germinal center (GC) reactions to immunization with Qbeta, Cr2(-/-) mice exhibited impaired antibody persistence paralleled by a strongly reduced development of bone marrow plasma cells. Surprisingly, antigen-specific memory B cells were essentially normal in these mice. In the absence of CD21-mediated costimulation, Qbeta-specific post-GC B cells failed to induce the transcriptional regulators Blimp-1 and XBP-1 driving plasma cell differentiation, and the antiapoptotic protein Bcl-2, which resulted in failure to generate the precursor population of long-lived plasma cells residing in the bone marrow. These results suggest that complement receptors maintain antibody responses by delivery of differentiation and survival signals to precursors of bone marrow plasma cells.
Assuntos
Células da Medula Óssea/imunologia , Proteínas de Ligação a DNA/biossíntese , Regulação da Expressão Gênica/imunologia , Proteínas Nucleares/biossíntese , Plasmócitos/imunologia , Receptores de Complemento 3d/imunologia , Proteínas Repressoras/biossíntese , Fatores de Transcrição/biossíntese , Allolevivirus/imunologia , Animais , Anticorpos Antivirais/imunologia , Formação de Anticorpos/genética , Formação de Anticorpos/imunologia , Antígenos CD , Antígenos Virais/imunologia , Apoptose/genética , Apoptose/imunologia , Células da Medula Óssea/citologia , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Sobrevivência Celular/genética , Sobrevivência Celular/imunologia , Proteínas de Ligação a DNA/imunologia , Regulação da Expressão Gênica/genética , Camundongos , Camundongos Knockout , Proteínas Nucleares/imunologia , Fator 1 de Ligação ao Domínio I Regulador Positivo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/imunologia , Receptores de Antígenos de Linfócitos B/genética , Receptores de Antígenos de Linfócitos B/imunologia , Receptores de Complemento 3d/genética , Fatores de Transcrição de Fator Regulador X , Proteínas Repressoras/imunologia , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Fatores de Transcrição/imunologia , Proteína 1 de Ligação a X-BoxRESUMO
An algorithm is presented for the detection of frequency contour sounds-whistles of dolphins and many other odontocetes, moans of baleen whales, chirps of birds, and numerous other animal and non-animal sounds. The algorithm works by tracking spectral peaks over time, grouping together peaks in successive time slices in a spectrogram if the peaks are sufficiently near in frequency and form a smooth contour over time. The algorithm has nine parameters, including the ones needed for spectrogram calculation and normalization. Finding optimal values for all of these parameters simultaneously requires a search of parameter space, and a grid search technique is described. The frequency contour detection method and parameter optimization technique are applied to the problem of detecting "boing" sounds of minke whales from near Hawaii. The test data set contained many humpback whale sounds in the frequency range of interest. Detection performance is quantified, and the method is found to work well at detecting boings, with a false-detection rate of 3% for the target missed-call rate of 25%. It has also worked well anecdotally for other marine and some terrestrial species, and could be applied to any species that produces a frequency contour, or to non-animal sounds as well.
Assuntos
Algoritmos , Baleia Anã/fisiologia , Processamento de Sinais Assistido por Computador , Vocalização Animal , Animais , Jubarte/fisiologia , Oceanos e Mares , Reprodutibilidade dos Testes , Espectrografia do Som , Especificidade da Espécie , Fatores de TempoRESUMO
The Candida albicans plasma membrane plays critical roles in growth and virulence and as a target for antifungal drugs. Three C. albicans genes that encode Bin-Amphiphysin-Rvs homology domain proteins were mutated to define their roles in plasma membrane function. The deletion of RVS161 and RVS167, but not RVS162, caused strong defects. The rvs161Delta mutant was more defective in endocytosis and morphogenesis than rvs167Delta, but both were strongly defective in polarizing actin patches. Other plasma membrane constituents were still properly localized, including a filipin-stained domain at the hyphal tips. An analysis of growth under different in vitro conditions showed that the rvs161Delta and rvs167Delta mutants grew less invasively in agar and also suggested that they have defects in cell wall synthesis and Rim101 pathway signaling. These mutants were also more resistant to the antimicrobial peptide histatin 5 but showed essentially normal responses to the drugs caspofungin and amphotericin. Surprisingly, the rvs161Delta mutant was more sensitive to fluconazole, whereas the rvs167Delta mutant was more resistant, indicating that these mutations cause overlapping but distinct effects on cells. The rvs161Delta and rvs167Delta mutants both showed greatly reduced virulence in mice. However, the mutants were capable of growing to high levels in kidneys. Histological analyses of infected kidneys revealed that these rvsDelta mutants grew in a large fungal mass that was walled off by leukocytes, rather than forming disseminated microabscesses as seen for the wild type. The diminished virulence is likely due to a combination of the morphogenesis defects that reduce invasive growth and altered cell wall construction that exposes proinflammatory components to the host immune system.
Assuntos
Candida albicans/patogenicidade , Endocitose , Proteínas Fúngicas/fisiologia , Actinas/química , Animais , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Feminino , Filipina/análise , Fluconazol/farmacologia , Histatinas/farmacologia , Hifas/crescimento & desenvolvimento , Camundongos , Camundongos Endogâmicos BALB C , Morfogênese , VirulênciaRESUMO
Marginal zone (MZ) B cells differ from follicular (FO) B cells in their functional, phenotypic and localization properties. It is still unclear whether B cells from the MZ compartment also have distinct or biased BCR specificities, recognizing only a limited number of conserved antigenic structures. To address the complexity of the immune response mounted by marginal zone B cells, we compared the antibody repertoire of murine MZ and FO B cells induced by immunization with two different virus-like particles (VLPs). Antibody sequences isolated from sorted VLP-specific MZ and FO B cells were similar in heavy chain V, D and J gene segment usage. Sequence analysis of CDR3 regions of antibodies from MZ and FO B cells also revealed no consistent difference in N nucleotide additions or CDR3 length. In contrast, somatic hypermutations were reduced in CDR regions of antibodies from MZ B cells compared to those from FO B cells. These results indicate that the response of MZ B cells to VLPs is clonotypically heterogeneous and suggest that the MZ B cell compartment is capable of generating variable and diverse antibody responses.
Assuntos
Diversidade de Anticorpos , Especificidade de Anticorpos , Subpopulações de Linfócitos B/imunologia , Vírion/imunologia , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/química , Anticorpos Antivirais/genética , Anticorpos Antivirais/imunologia , Regiões Determinantes de Complementaridade , Feminino , Genes de Cadeia Pesada de Imunoglobulina/genética , Imunização , Cadeias Pesadas de Imunoglobulinas/química , Cadeias Pesadas de Imunoglobulinas/genética , Cadeias Pesadas de Imunoglobulinas/imunologia , Região Variável de Imunoglobulina/química , Região Variável de Imunoglobulina/genética , Região Variável de Imunoglobulina/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Hipermutação Somática de ImunoglobulinaRESUMO
The plant plasma membrane is now known to be a more sophisticated structure than was previously thought. Sebastien Mongrand et al. and Georg Borner et al. have isolated specific plasma membrane microdomains ('lipid rafts') that are enriched in sterols and sphingolipids. These rafts contain distinct sets of proteins and might help to explain how plasma membrane proteins are positioned in certain parts of the cell to function in development and signalling.
Assuntos
Microdomínios da Membrana/fisiologia , Plantas/química , Membrana Celular/química , Membranas Intracelulares/química , Proteínas de Plantas/metabolismo , Plantas/ultraestruturaRESUMO
Military sonar has the potential to negatively impact marine mammals. To investigate factors affecting behavioral disruption in California sea lions (Zalophus californianus), fifteen sea lions participated in a controlled exposure study using a simulated tactical sonar signal (1 s duration, 3250-3450 Hz) as a stimulus. Subjects were placed into groups of three and each group received a stimulus exposure of 125, 140, 155, 170, or 185 dB re: 1 µPa (rms). Each subject was trained to swim across an enclosure, touch a paddle, and return to the start location. Sound exposures occurred at the mid-point of the enclosure. Control and exposure sessions were run consecutively and each consisted of ten, 30-s trials. The occurrence and severity of behavioral responses were used to create acoustic dose-response and dose-severity functions. Age of the subject significantly affected the dose-response relationship, but not the dose-severity relationship. Repetitive exposures did not affect the dose-response relationship.
Assuntos
Comportamento Animal , Leões-Marinhos/fisiologia , Som , Estimulação Acústica , Fatores Etários , Animais , RuídoRESUMO
Hypochlorous acid (HOCl), produced in inflammatory conditions by the enzyme myeloperoxidase, and its anion hypochlorite (OCl(-)) exist in vivo at almost equal concentrations. Their reactions with hyaluronan and heparin (as a model for sulfated glycosaminoglycans in the extracellular matrix) have been studied as a function of pH. The major product in these reactions is the chloramide derivative of the glycosaminoglycans. Spectral, chloramide yield, and kinetic measurements show sharply contrasting behavior of heparin and hyaluronan and the data allow the calculation of second-order rate constants for the reactions of both HOCl and OCl(-) for all reaction pathways leading to the formation of chloramides and also oxidation products. By comparison with hyaluronan, it can be demonstrated that both N-sulfate and O-sulfate groups in heparin influence the proportions of these pathways in this glycosaminoglycan. Evidence is also given for further oxidation pathways involving a reaction of HOCl with the chloramide product of hyaluronan but not with heparin. The significance of these results for the mechanisms of inflammation, particularly for fragmentation of extracellular matrix glycosaminoglycans, is discussed.
Assuntos
Cloro/química , Halogenação , Heparina/química , Ácido Hialurônico/química , Ácido Hipocloroso/química , Sulfatos/química , Ânions/química , Cinética , OxirreduçãoRESUMO
Reliable estimation of the size or density of wild animal populations is very important for effective wildlife management, conservation and ecology. Currently, the most widely used methods for obtaining such estimates involve either sighting animals from transect lines or some form of capture-recapture on marked or uniquely identifiable individuals. However, many species are difficult to sight, and cannot be easily marked or recaptured. Some of these species produce readily identifiable sounds, providing an opportunity to use passive acoustic data to estimate animal density. In addition, even for species for which other visually based methods are feasible, passive acoustic methods offer the potential for greater detection ranges in some environments (e.g. underwater or in dense forest), and hence potentially better precision. Automated data collection means that surveys can take place at times and in places where it would be too expensive or dangerous to send human observers. Here, we present an overview of animal density estimation using passive acoustic data, a relatively new and fast-developing field. We review the types of data and methodological approaches currently available to researchers and we provide a framework for acoustics-based density estimation, illustrated with examples from real-world case studies. We mention moving sensor platforms (e.g. towed acoustics), but then focus on methods involving sensors at fixed locations, particularly hydrophones to survey marine mammals, as acoustic-based density estimation research to date has been concentrated in this area. Primary among these are methods based on distance sampling and spatially explicit capture-recapture. The methods are also applicable to other aquatic and terrestrial sound-producing taxa. We conclude that, despite being in its infancy, density estimation based on passive acoustic data likely will become an important method for surveying a number of diverse taxa, such as sea mammals, fish, birds, amphibians, and insects, especially in situations where inferences are required over long periods of time. There is considerable work ahead, with several potentially fruitful research areas, including the development of (i) hardware and software for data acquisition, (ii) efficient, calibrated, automated detection and classification systems, and (iii) statistical approaches optimized for this application. Further, survey design will need to be developed, and research is needed on the acoustic behaviour of target species. Fundamental research on vocalization rates and group sizes, and the relation between these and other factors such as season or behaviour state, is critical. Evaluation of the methods under known density scenarios will be important for empirically validating the approaches presented here.
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Acústica , Ecossistema , Modelos Biológicos , Animais , Conservação dos Recursos Naturais , Coleta de Dados , Densidade DemográficaRESUMO
Hypochlorous acid and its conjugate base, hypochlorite ions, produced under inflammatory conditions, may produce chloramides of glycosaminoglycans, these being significant components of the extracellular matrix (ECM). This may occur through the binding of myeloperoxidase directly to the glycosaminoglycans. The N-Cl group in the chloramides is a potential selective target for both reducing and oxidizing radicals, leading possibly to more efficient and damaging fragmentation of these biopolymers relative to the parent glycosaminoglycans. To investigate the effect of the N-Cl group, we used ionizing radiation to produce quantifiable concentrations of the reducing radicals, hydrated electron and superoxide radical, and also of the oxidizing radicals, hydroxyl, carbonate, and nitrogen dioxide, all of which were reacted with hyaluronan and heparin and their chloramides in this study. PAGE gels calibrated for molecular weight allowed the consequent fragmentation efficiencies of these radicals to be calculated. Hydrated electrons were shown to produce fragmentation efficiencies of 100 and 25% for hyaluronan chloramide (HACl) and heparin chloramide (HepCl), respectively. The role of the sulfate group in heparin in the reduction of fragmentation can be rationalized using mechanisms proposed by M.D. Rees et al. (J. Am. Chem. Soc.125:13719-13733; 2003), in which the initial formation of an amidyl radical leads rapidly to a C-2 radical on the glucosamine moiety. This is 100% efficient at causing glycosidic bond breakage in HACl but only 25% efficient in HepCl, the role of the sulfate group being to favor the nonfragmentary routes for the C-2 radical. The weaker reducing agent, the superoxide radical, did not cause fragmentation of either HACl or HepCl although kinetic reactivity had been demonstrated in earlier studies. Experiments using the oxidizing radicals, hydroxyl and carbonate, both potential in vivo species, showed significant increases in fragmentation efficiencies for both HACl and HepCl, relative to the parent molecules. The carbonate radical was shown to be involved in site-specific reactions at the N-Cl groups, reacting via abstraction of Cl, to produce the same amidyl radical produced by one-electron reductants such as the hydrated electron. As for the hydrated electrons, the data support fragmentation efficiencies of 100 and 29% for reaction of carbonate radicals at N-Cl for HACl and HepCl, respectively. For the weaker oxidant, nitrogen dioxide, no fragmentation was observed, probably because of a low kinetic reactivity and low reduction potential. It seems likely therefore that the N-Cl group can direct damage to extracellular matrix glycosaminoglycan chloramides, which may be produced under inflammatory conditions. The in vivo species, the carbonate radical, is also much more likely to be site-specific in its reactions with such components of the ECM than the hydroxyl radical.
Assuntos
Matriz Extracelular/metabolismo , Glicosaminoglicanos/metabolismo , Heparina/metabolismo , Ácido Hialurônico/metabolismo , Inflamação/metabolismo , Animais , Matriz Extracelular/química , Radicais Livres/metabolismo , Radicais Livres/toxicidade , Glicosaminoglicanos/química , Heparina/química , Humanos , Ácido Hialurônico/química , Oxirredução , Estresse Oxidativo/fisiologiaRESUMO
Protective Ab levels can be maintained for years upon infection or vaccination. In this study, we studied the duration of Ab responses as a function of the life span of plasma cells and tested the role of persisting Ag in maintaining B cell memory. Our analysis of B cell responses induced in mice immunized with virus-like particles demonstrates the following: 1) Ab titers are long-lived, but decline continuously with a t(1/2) of approximately 80 days, which corresponds to the life span of plasma cells; 2) the germinal center (GC) reaction, which lasts for up to 100 days, is dependent on Ag associated with follicular dendritic cells; and 3) early GCs produce massive numbers of plasma and memory B cell precursors, whereas the late Ag-dependent GCs are dispensable for the maintenance of Ab levels and B cell memory.