RESUMEN
Nod-like receptor family pyrin domain-containing 3 (NLRP3) is a cytosolic innate immune receptor that senses organelle dysfunction induced by various stimuli, such as infectious, environmental, metabolic and drug stresses. Upon activation, NLRP3 forms an inflammasome with its adaptor protein apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and caspase-1, to trigger the release of inflammatory cytokines. The development of effective anti-inflammatory drugs targeting the NLRP3 inflammasome is in high demand as its aberrant activation often causes inflammatory diseases. Here, we found that nanaomycin A (NNM-A), a quinone-based antibiotic isolated from Streptomyces, effectively inhibited NLRP3 inflammasome-mediated inflammatory responses induced by imidazoquinolines, including imiquimod. Interestingly, its epoxy derivative nanaomycin E (NNM-E) showed a comparable inhibitory effect against the NLRP3 inflammasome-induced release of interleukin (IL)-1ß and IL-18 from macrophages, with a much lower toxicity than NNM-A. NNM-E inhibited ASC oligomerization and caspase-1 cleavage, both of which are hallmarks of NLRP3 inflammasome activation. NNM-E reduced mitochondrial damage and the production of reactive oxygen species, thereby preventing the activation of the NLRP3 inflammasome. NNM-E treatment markedly alleviated psoriasis-like skin inflammation induced by imiquimod. Collectively, NNM-E inhibits NLRP3 inflammasome activation by preventing mitochondrial dysfunction with little toxicity and showed an anti-inflammatory effect in vivo. Thus, NNM-E could be a potential lead compound for developing effective and safe anti-inflammatory agents for the treatment of NLRP3 inflammasome-mediated inflammatory diseases.
Asunto(s)
Inflamasomas , Proteína con Dominio Pirina 3 de la Familia NLR , Caspasa 1/metabolismo , Imiquimod/metabolismo , Imiquimod/farmacología , Interleucina-1beta/metabolismo , Mitocondrias/metabolismo , NaftoquinonasRESUMEN
BACKGROUND AND AIMS: Dipeptidyl peptidase-4 (DPP-4) inhibitors have been reported to suppress atherosclerosis progression in atherosclerotic mouse models through unclear mechanisms. In this study, we investigated the effect of the DPP-4 inhibitor, linagliptin, on macrophage polarization in vitro and in vivo. METHODS: Mouse bone marrow macrophages (BMMs) were used in in vitro assays. High fat diet (HFD)-fed Apoe-/- mice were treated orally with linagliptin (10 mg/kg-1â¢day-1) or a vehicle (water) control. RESULTS: In in vitro assays using BMMs, treatment with LPS and IFNγ decreased the mRNA-expression levels of alternatively activated macrophage (M2) markers, and linagliptin treatment prevented these reductions. The mRNA levels of M2 markers and the number of M2 macrophages in the aorta were higher in linagliptin groups than in control groups. Linagliptin decreased the size of atherosclerotic lesions in HFD-fed Apoe-/- mice. Interestingly, inflammatory stimulation increased DPP-4 expression, and linagliptin suppressed these effects in BMMs. Treatment with DPP-4 small-interfering RNA (siRNA) reproduced linagliptin-mediated alteration of M2 polarization. CONCLUSIONS: Linagliptin increased M2 macrophage polarization by inhibiting DPP-4 expression and activity. These findings may indicate the beneficial effects of DPP-4 inhibitors on the progression of diabetic macrovascular complications.
Asunto(s)
Antiinflamatorios/química , Dipeptidil Peptidasa 4/metabolismo , Inhibidores de la Dipeptidil-Peptidasa IV/química , Inflamación/tratamiento farmacológico , Linagliptina/química , Animales , Antiinflamatorios/farmacología , Aorta/metabolismo , Aterosclerosis/tratamiento farmacológico , Células de la Médula Ósea/efectos de los fármacos , Dieta Alta en Grasa , Dipeptidil Peptidasa 4/genética , Inhibidores de la Dipeptidil-Peptidasa IV/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Linagliptina/farmacología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Animales , ARN Mensajero/metabolismo , ARN Interferente Pequeño/metabolismoRESUMEN
While common semantic representations for individual words across languages have been identified, a common meaning system at sentence-level has not been determined. In this study, fMRI was used to investigate whether an across-language sentence comprehension system exists. Chinese-Japanese bilingual participants (n = 32) were asked to determine whether two consecutive stimuli were related (coherent) or not (incoherent) to the same event. Stimuli were displayed with three different modalities (Chinese written sentences, Japanese written sentences, and pictures). The behavioral results showed no significant difference in accuracy and response times among the three modalities. Multi-voxel pattern analysis (MVPA) of fMRI data was used to classify the semantic relationship (coherent or incoherent) across the stimulus modalities. The classifier was first trained to determine coherency within Chinese sentences and then tested with Japanese sentences, and vice versa. A whole-brain searchlight analysis revealed significant above-chance classification accuracy across Chinese and Japanese sentences in the supramarginal gyrus (BA 40), extending into the angular gyrus (BA 39) as well as the opercular (BA 44) and triangular (BA 45) parts of the inferior frontal gyrus in the left hemisphere (cluster-level FWE corrected p < 0.05). Significant above-chance classification accuracy was also found across Japanese sentences and pictures in the supramarginal (BA 40) and angular gyrus (BA 39). These results indicate that a common meaning system for sentence processing across languages and modalities exists, and it involves the left inferior parietal gyrus.
RESUMEN
We investigated the effects of long-term training on the neural representation of individual finger movements in the primary sensorimotor cortex. One group of participants (trained group) included subjects trained in playing the piano (mean years of experienceâ¯=â¯17.9; rangeâ¯=â¯9-26; nâ¯=â¯20). The other group of participants (novice group) had no prior experience (nâ¯=â¯20). All participants performed finger-tapping movements using either of the four digits of the hand (index, middle, ring, and little fingers). Functional magnetic resonance imaging (fMRI) was used to analyze the spatial activation patterns elicited by individual finger movements. Subsequently, we tried to classify the finger that was being moved using a multi-voxel pattern analysis (MVPA). Our results showed significantly higher-than-chance classification accuracies in both primary motor cortex (M1) and somatosensory cortex (S1) contralateral to the hand. We also found significantly lower classification accuracies for both hands in the trained group compared with the novice group in M1, without significant differences in the average signal changes and the number of activated voxels for individual fingers or overlap between digits. Representational similarity analysis (RSA) also demonstrated the differences in similarity patterns of activations between the trained and novice groups in M1. Our results indicate the modulation of neural representations of individual finger movements of M1 due to long-term training.
Asunto(s)
Actividad Motora/fisiología , Corteza Motora/fisiología , Desempeño Psicomotor/fisiología , Adulto , Mapeo Encefálico , Femenino , Dedos , Humanos , Imagen por Resonancia Magnética , Masculino , Análisis Multivariante , Práctica Psicológica , Corteza Sensoriomotora/fisiología , Adulto JovenRESUMEN
BACKGROUND AND AIMS: Local macrophage proliferation is linked to enhanced atherosclerosis progression. Our previous study found that troglitazone, a thiazolidinedione (TZD), suppressed oxidized low-density lipoprotein (Ox-LDL)-induced macrophage proliferation. However, its effects and mechanisms are unclear. Therefore, we investigated the effects of pioglitazone, another TZD, on macrophage proliferation. METHODS: Normal chow (NC)- or high-fat diet (HFD)-fed apolipoprotein E-deficient (Apoe-/-) mice were treated orally with pioglitazone (10â¯mg/kg/day) or vehicle (water) as a control. Mouse peritoneal macrophages were used in in vitro assays. RESULTS: Atherosclerosis progression was suppressed in aortic sinuses of pioglitazone-treated Apoe-/- mice, which showed fewer proliferating macrophages in plaques. Pioglitazone suppressed Ox-LDL-induced macrophage proliferation in a dose-dependent manner. However, treatment with peroxisome proliferator-activated receptor-γ (PPARγ) siRNA ameliorated pioglitazone-induced suppression of macrophage proliferation. Low concentrations (less than 100⯵mol/L) of pioglitazone, which can suppress macrophage proliferation, activated PPARγ in macrophages, but did not induce macrophage apoptosis. Pioglitazone treatment did not induce TUNEL-positive cells in atherosclerotic plaques of aortic sinuses in Apoe-/- mice. CONCLUSIONS: Pioglitazone suppressed macrophage proliferation through PPARγ without inducing macrophage apoptosis. These findings imply that pioglitazone could prevent macrovascular complications in diabetic individuals.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Macrófagos/citología , Macrófagos/efectos de los fármacos , PPAR gamma/fisiología , Pioglitazona/farmacología , Animales , Apolipoproteínas E/deficiencia , Aterosclerosis/prevención & control , Masculino , Ratones , Ratones Endogámicos C57BL , Pioglitazona/uso terapéuticoRESUMEN
An unusual combination of a laboratory experiment and in situ measurement of pressure fluctuations during an earthquake allows us to resolve some uncertainties in bottom pressure recorders (BPRs). In situ BPRs are usually contaminated by seismic waves during earthquakes; thus uncertainty still remains in the data obtained from BPRs. We examine in situ BPR data together with pressure variations produced by a dead weight (a pressure standard) in a laboratory experiment during an earthquake. The features recorded by the in situ BPRs are analysed as part of the overall experiment. We demonstrated that a 10-kg dead weight on a piston-cylinder across an area of 10 mm2 is capable of reproducing pressure fluctuations at a depth of 1000 m in the water column. The experiment also indicates that the internal mechanics of BPRs are isolated from incident seismic waves, suggesting that BPRs measure true in situ pressures without instrumentally induced disturbances. This constitutes the first instance in which pressure fluctuations recorded by in situ BPRs during an earthquake were reproduced using a pressure standard in the laboratory.
RESUMEN
OBJECTIVE: Macrophages play a central role in various stages of atherosclerotic plaque formation and progression. The local macrophages reportedly proliferate during atherosclerosis, but the pathophysiological significance of macrophage proliferation in this context remains unclear. Here, we investigated the involvement of local macrophage proliferation during atherosclerosis formation and progression using transgenic mice, in which macrophage proliferation was specifically suppressed. APPROACH AND RESULTS: Inhibition of macrophage proliferation was achieved by inducing the expression of cyclin-dependent kinase inhibitor 1B, also known as p27kip, under the regulation of a scavenger receptor promoter/enhancer. The macrophage-specific human p27kip Tg mice were subsequently crossed with apolipoprotein E-deficient mice for the atherosclerotic plaque study. Results showed that a reduced number of local macrophages resulted in marked suppression of atherosclerotic plaque formation and inflammatory response in the plaque. Moreover, fewer local macrophages in macrophage-specific human p27kip Tg mice helped stabilize the plaque, as evidenced by a reduced necrotic core area, increased collagenous extracellular matrix, and thickened fibrous cap. CONCLUSIONS: These results provide direct evidence of the involvement of local macrophage proliferation in formation and progression of atherosclerotic plaques and plaque stability. Thus, control of macrophage proliferation might represent a therapeutic target for treating atherosclerotic diseases.
Asunto(s)
Aorta/patología , Aortitis/prevención & control , Aterosclerosis/prevención & control , Proliferación Celular , Activación de Macrófagos , Macrófagos Peritoneales/patología , Placa Aterosclerótica , Animales , Aorta/metabolismo , Aortitis/genética , Aortitis/metabolismo , Aortitis/patología , Aterosclerosis/genética , Aterosclerosis/metabolismo , Aterosclerosis/patología , Células Cultivadas , Colágeno/metabolismo , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Modelos Animales de Enfermedad , Fibrosis , Mediadores de Inflamación/metabolismo , Macrófagos Peritoneales/metabolismo , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados para ApoE , Ratones Transgénicos , Necrosis , Transducción de SeñalRESUMEN
Sanu Vengasseril Francis, Shuhei Nishida, and Sivasankaran Bijoy Nandan (2018) The neustonic copepods of the family Pontellidae - Pontella spinipes Giesbrecht, 1889 and P. diagonalis Wilson, 1950, both first described on the basis of female specimens exhibit very similar morphology and overlapping geographic ranges in the Indian Ocean. While several taxonomists have described males of each species, there has been no definitive evidence for female-male matching (link female and male of the same species) in the two species. In the present study, an analysis of mitochondrial cytochrome c oxidase subunit I (mtCOI) sequences in the specimens collected from the Arabian Sea revealed that female P. spinipes Giesbrecht, 1889, and male P. diagonalis sensu Silas and Pillai (1973) are genetically identical, providing evidence that the latter is actually P. spinipes. These findings emphasize that it is necessary to re-examine the female-male correspondence of other related species, formerly based on morphology alone, using molecular-genetic analysis as applied in the present study.
RESUMEN
Hepatitis C-associated osteosclerosis (HCAO), a very rare disorder in which an extremely rapid bone turnover occurs and results in osteosclerosis, was acknowledged in 1990s as a new clinical entity with the unique bone disorder and definite link to chronic type C hepatitis, although the pathogenesis still remains unknown. Affected patients suffer from excruciating deep bone pains. We report the 19th case of HCAO with diagnosis confirmed by bone biopsy, and treated initially with a bisphosphonate, next with corticosteroids and finally with direct acting antivirals (DAA: sofosbuvir and ribavirin) for HCV infection. Risedronate, 17.5 mg/day for 38 days, did not improve the patient's symptoms or extremely elevated levels of bone markers, which indicated hyper-bone-formation and coexisting hyper-bone-resorption in the patient. Next, intravenous methylprednisolone pulse therapy followed by high-dose oral administration of prednisolone evidently improved them. DAA therapy initiated after steroid therapy successfully achieved sustained virological response, but no additional therapeutic effect on them was observed. Our results strongly suggested that the underlying immunological alteration is the crucial key to clarify the pathogenesis of HCAO. Bone mineral density of lumbar vertebrae of the patient was increased by 14% in four-month period of observation. Clarification of the mechanisms that develop osteosclerosis in HCAO might lead to a new therapeutic perspective for osteoporosis. LEARNING POINTS: HCAO is an extremely rare bone disorder, which occurs exclusively in patients affected with HCV, of which only 18 cases have been reported since 1992 and pathogenesis still remains unclear.Pathophysiology of HCAO is highly accelerated rates of both bone formation and bone resorption, with higher rate of formation than that of resorption, which occur in general skeletal leading to the diffuse osteosclerosis with severe bone pains.Steroid therapy including intravenous pulse administration in our patient evidently ameliorated his bone pains and reduced elevated values of bone markers. This was the first successful treatment for HCAO among cases reported so far and seemed to propose a key to solve the question for its pathogenesis.The speed of increase in the bone mineral content of the patient was very high, suggesting that clarification of the mechanism(s) might lead to the development of a novel therapy for osteoporosis.
RESUMEN
Lipids of Neocalanus cristatus and Eucalanus bungii (C3 to adults), collected in March, May, and December from various depths (0-2000m) were studied in the Oyashio region, western North Pacific. Total lipid and wax ester contents of younger N. cristatus stages increased during the development, being higher in May than in March and December. Major fatty acids of younger N. cristatus were 16:0, 20:5(n-3), and 22:6(n-3) and the dominant alcohols were 16:0, 16:1(n-7), 20:1(n-9)/(n-11) and 22:1(n-11). The energy-rich 20:1 and 22:1 moieties increased from the younger to the adult stages showing the importance of lipid biosynthesis which may be advantageous for successful overwintering and reproduction at depth. The 16:4(n-1) fatty acid, characteristic of a diatom diet increased in May, particularly in the younger stages. Our results suggest that the diatom-dominated feeding mode of younger N. cristatus during the spring bloom is important for an effective accumulation of wax esters. In contrast to N. cristatus, E. bungii accumulated substantial amounts of triacylglycerols. The total lipid and triacylglycerol content increased slightly toward the older developmental stages. The major fatty acids were 16:0, 16:1(n-7), 18:1(n-9) and (n-7), and 20:5(n-3). There was no evidence of developmental or seasonal changes in the fatty acid composition. The differences in the lipid storage modes of both copepods via wax esters or triacylglycerols are species-specific but their fatty acid compositions varied according to diet and developmental stage, especially in N. cristatus. These lipid characteristics are discussed in relation to reproduction, feeding modes, diapause and overwintering strategies.
Asunto(s)
Alcoholes/química , Copépodos/química , Ácidos Grasos/química , Lípidos/química , Océano Pacífico , Alcoholes/metabolismo , Animales , Copépodos/metabolismo , Metabolismo Energético , Ácidos Grasos/metabolismo , Metabolismo de los Lípidos , Estaciones del AñoRESUMEN
A new species of the calanoid copepod genus Pseudodiaptomus was collected from the Prasae River Estuary, Rayong Province, on the eastern coast of the Gulf of Thailand. This species is definitely assigned to the lobus species group sensu Walter (1986a). The female of the new species differs from other congeners in the elongate genital double-somite with a blunt process ventrally and the second urosomite about 2.54 times as long as wide. The male is also easily distinguished from other congeners by the structure of the right fifth leg. The present new species is a euryhaline species and occurred in brackish waters with salinity ranging from 0.7 to 23.3. Its breeding season may be from June to October, as indicated by the presence of egg-sacs.
RESUMEN
Here we describe the application of a recently developed high-resolution microcantilever biosensor resonating at the air-liquid interface for the continuous detection of antigen-antibody and enzyme-substrate interactions. The cantilever at the air-liquid interface demonstrated 50% higher quality factor and a 5.7-fold increase in signal-to-noise-ratio (SNR) compared with one immersed in the purified water. First, a label-free detection of a low molecular weight protein (insulin, 5.8 kDa) in physiological concentration was demonstrated. The liquid facing side of the cantilever was functionalized by coating its surface with insulin antibodies, while the opposite side was exposed to air. The meniscus membrane at the micro-slit around the cantilever sustained the liquid in the microchannel. After optimizing the process of surface functionalization, the resonance frequency shift was successfully measured for insulin solutions of 0.4, 2.0, and 6.3 ng ml(-1). To demonstrate additional application of the device for monitoring enzymatic protein degradation, the liquid facing microcantilever surface was coated with human recombinant SOD1 (superoxide dismutase 1) and exposed to various concentrations of proteinase K solution, and the kinetics of the SOD1 digestion was continuously monitored. The results showed that it is a suitable tool for sensitive protein detection and analysis.
Asunto(s)
Anticuerpos/química , Técnicas Biosensibles/métodos , Insulina/análisis , Técnicas Analíticas Microfluídicas/métodos , Superóxido Dismutasa/química , Técnicas Biosensibles/instrumentación , Humanos , Técnicas para Inmunoenzimas , Insulina/química , Insulina/metabolismo , Técnicas Analíticas Microfluídicas/instrumentación , Sensibilidad y Especificidad , Superóxido Dismutasa-1RESUMEN
We have developed a highly mass-sensitive cantilever resonating at the interface of air and liquid. The cantilever is applicable as a biosensor by measuring its resonance frequency shift associated with the selective trapping of target molecules. One surface of the cantilever facing to the liquid is functionalized for label-free detection, while the opposite side is exposed to air to improve the resonance characteristics, such as the quality factor. The quality factor at resonance is 15, which is 50% higher than the same cantilever in liquid. The beam was excited through the photothermal effect of a power modulated laser and detected by laser Doppler velocimetry. Due to the proposed configuration, the signal-to-noise-ratio is 5.7 times larger than the completely submerged case. A micro-slit around the cantilever separates the air and liquid phases at a meniscus. We analyzed the cantilever motion including the meniscus membrane, and examined the effect of surface tension by applying various solutions. A slit width of 6 µm was found to give the best performance within the few prototypes. We measured the covalent immobilization of antibody molecules on a cantilever surface for three different concentrations: 20, 40, and 80 µg ml(-1). The kinetics measured by both resonance frequency shift of the cantilever and fluorescent intensity showed good agreement.
Asunto(s)
Aire , Técnicas Biosensibles/métodos , Técnicas Analíticas Microfluídicas/instrumentación , Anticuerpos Inmovilizados/química , Técnicas Biosensibles/instrumentación , Cinética , Flujometría por Láser-Doppler , Relación Señal-Ruido , Dióxido de Silicio/química , Tensión SuperficialRESUMEN
To understand marine biodiversity in Japanese waters, we have compiled information on the marine biota in Japanese waters, including the number of described species (species richness), the history of marine biology research in Japan, the state of knowledge, the number of endemic species, the number of identified but undescribed species, the number of known introduced species, and the number of taxonomic experts and identification guides, with consideration of the general ocean environmental background, such as the physical and geological settings. A total of 33,629 species have been reported to occur in Japanese waters. The state of knowledge was extremely variable, with taxa containing many inconspicuous, smaller species tending to be less well known. The total number of identified but undescribed species was at least 121,913. The total number of described species combined with the number of identified but undescribed species reached 155,542. This is the best estimate of the total number of species in Japanese waters and indicates that more than 70% of Japan's marine biodiversity remains un-described. The number of species reported as introduced into Japanese waters was 39. This is the first attempt to estimate species richness for all marine species in Japanese waters. Although its marine biota can be considered relatively well known, at least within the Asian-Pacific region, considering the vast number of different marine environments such as coral reefs, ocean trenches, ice-bound waters, methane seeps, and hydrothermal vents, much work remains to be done. We expect global change to have a tremendous impact on marine biodiversity and ecosystems. Japan is in a particularly suitable geographic situation and has a lot of facilities for conducting marine science research. Japan has an important responsibility to contribute to our understanding of life in the oceans.
Asunto(s)
Biodiversidad , Agua de Mar , Animales , Clasificación , Sistemas de Información , Japón , Océanos y MaresRESUMEN
Trace elements (TEs) and stable isotope ratios (delta(15)N and delta(13)C) were analyzed in fish from deep-water of the Sulu Sea, the Celebes Sea and the Philippine Sea. Concentrations of V and Pb in pelagic fish from the Sulu Sea were higher than those from the Celebes Sea, whereas the opposite trend was observed for delta(13)C. High concentrations of Zn, Cu and Ag were found in non-migrant fish in deep-water, while Rb level was high in fish which migrate up to the epipelagic zone, probably resulting from differences in background levels of these TEs in each water environment or function of adaptation to deep-water by migrant and non-migrant species. Arsenic level in the Sulu Sea fish was positively correlated with delta(15)N, indicating biomagnification of arsenic. To our knowledge, this is the first study on relationship between diel vertical migration and TE accumulation in deep-water fish.
Asunto(s)
Isótopos de Carbono/análisis , Ecosistema , Peces/metabolismo , Isótopos de Nitrógeno/análisis , Oligoelementos/análisis , Oligoelementos/metabolismo , Migración Animal/fisiología , Animales , Contaminación de Alimentos/análisis , Geografía , Humanos , Océanos y MaresRESUMEN
The complete nucleotide sequences of the mitochondrial genomes were determined for the three pelagic chaetognaths, Sagitta nagae, Sagitta decipiens, and Sagitta enflata. The mitochondrial genomes of these species which were 11,459, 11,121, and 12,631bp in length, respectively, contained 14 genes (11 protein-coding genes, one transfer RNA gene, and two ribosomal RNA genes), and were found to have lost 23 genes that are present in the typical metazoan mitochondrial genome. The same mitochondrial genome contents have been reported from the benthic chaetognaths belonging to the family Spadellidae, Paraspadella gotoi and Spadella cephaloptera. Within the phylum Chaetognatha, Sagitta and Spadellidae are distantly related, suggesting that the gene loss occurred in the ancestral species of the phylum. The gene orders of the three Sagitta species are markedly different from those of the other non-Chaetognatha metazoans. In contrast to the region with frequent gene rearrangements, no gene rearrangements were observed in the gene cluster encoding COII-III, ND1-3, srRNA, and tRNA(met). Within this conserved gene cluster, gene rearrangements were not observed in the three Sagitta species or between the Sagitta and Spadellidae species. The gene order of this cluster was also assumed to be the ancestral state of the phylum.
Asunto(s)
Anélidos/genética , Ecosistema , Genoma Mitocondrial/genética , Animales , Composición de Base/genética , Secuencia de Bases , Codón Iniciador/genética , Codón de Terminación/genética , ADN Mitocondrial/genética , Orden Génico/genética , Reordenamiento Génico/genética , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Sistemas de Lectura Abierta/genética , ARN Ribosómico/química , ARN Ribosómico/genética , ARN de Transferencia/química , ARN de Transferencia/genéticaRESUMEN
BACKGROUND: Oceans cover more than 70% of the earth's surface and are critical for the homeostasis of the environment. Among the components of the ocean ecosystem, zooplankton play vital roles in energy and matter transfer through the system. Despite their importance, understanding of zooplankton biodiversity is limited because of their fragile nature, small body size, and the large number of species from various taxonomic phyla. Here we present the results of single-gene zooplankton community analysis using a method that determines a large number of mitochondrial COI gene sequences from a bulk zooplankton sample. This approach will enable us to estimate the species richness of almost the entire zooplankton community. RESULTS: A sample was collected from a depth of 721 m to the surface in the western equatorial Pacific off Pohnpei Island, Micronesia, with a plankton net equipped with a 2-m2 mouth opening. A total of 1,336 mitochondrial COI gene sequences were determined from the cDNA library made from the sample. From the determined sequences, the occurrence of 189 species of zooplankton was estimated. BLASTN search results showed high degrees of similarity (>98%) between the query and database for 10 species, including holozooplankton and merozooplankton. CONCLUSION: In conjunction with the Census of Marine Zooplankton and Barcode of Life projects, single-gene zooplankton community analysis will be a powerful tool for estimating the species richness of zooplankton communities.
Asunto(s)
Biodiversidad , Genes Mitocondriales , Zooplancton/genética , Animales , ADN Mitocondrial/genética , Complejo IV de Transporte de Electrones/genética , Biblioteca de Genes , Micronesia , Océano Pacífico , Filogenia , ARN Mensajero/genética , Análisis de Secuencia de ADN , Zooplancton/clasificaciónRESUMEN
The authors present an optically based method combining photothermal excitation and laser Doppler velocimetry of higher cantilever vibration modes for dynamic atomic force microscopy in liquid. The frequency spectrum of a silicon cantilever measured in water over frequencies ranging up to 10 MHz shows that the method allows us to excite and detect higher modes, from fundamental to fifth flexural, without enhancing spurious resonances. By reducing the tip oscillation amplitude using higher modes, the average tip-sample force gradient due to chemical bonds is effectively increased to achieve high-spatial-resolution imaging in liquid. The method's performance is demonstrated by atomic resolution imaging of a mica surface in water obtained using the second flexural mode with a small tip amplitude of 99 pm; individual atoms on the surface with small height differences of up to 60 pm are clearly resolved.
RESUMEN
We determined the nearly complete DNA sequence of the mitochondrial genome of Antarctic krill Euphausia superba (Crustacea: Malacostraca), one of the most ecologically and commercially important zooplankters in Antarctic waters. All of the genome sequences were purified by gene amplification using long polymerase chain reaction (PCR), and the products were subsequently used as templates for either direct sequencing using a primer-walking strategy or nested PCR with crustacea-versatile primers. Although we were unable to determine a portion of the genome owing to technical difficulties, the sequenced position, 14,606 bp long, contained all of the 13 protein-coding genes, 19 of the 22 transfer RNA genes, and the large subunit as well as a portion of the small subunit ribosomal RNA genes. Gene rearrangement was observed for 3 transfer RNA genes (tRNACys, tRNATyr, and tRNATrp) and the 2 leucine tRNA genes.