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BACKGROUND: The complex relationship between atrial fibrillation (AF) and type 2 diabetes mellitus (T2DM) suggests a potential role for epicardial adipose tissue (EAT) that requires further investigation. This study employs bioinformatics and experimental approaches to clarify EAT's role in linking T2DM and AF, aiming to unravel the biological mechanisms involved. METHOD: Bioinformatics analysis initially identified common differentially expressed genes (DEGs) in EAT from T2DM and AF datasets. Pathway enrichment and network analyses were then performed to determine the biological significance and network connections of these DEGs. Hub genes were identified through six CytoHubba algorithms and subsequently validated biologically, with further in-depth analyses confirming their roles and interactions. Experimentally, db/db mice were utilized to establish a T2DM model. AF induction was executed via programmed transesophageal electrical stimulation and burst pacing, focusing on comparing the incidence and duration of AF. Frozen sections and Hematoxylin and Eosin (H&E) staining illuminated the structures of the heart and EAT. Moreover, quantitative PCR (qPCR) measured the expression of hub genes. RESULTS: The study identified 106 DEGs in EAT from T2DM and AF datasets, underscoring significant pathways in energy metabolism and immune regulation. Three hub genes, CEBPZ, PAK1IP1, and BCCIP, emerged as pivotal in this context. In db/db mice, a marked predisposition towards AF induction and extended duration was observed, with HE staining verifying the presence of EAT. Additionally, qPCR validated significant changes in hub genes expression in db/db mice EAT. In-depth analysis identified 299 miRNAs and 33 TFs as potential regulators, notably GRHL1 and MYC. GeneMANIA analysis highlighted the hub genes' critical roles in stress responses and leukocyte differentiation, while immune profile correlations highlighted their impact on mast cells and neutrophils, emphasizing the genes' significant influence on immune regulation within the context of T2DM and AF. CONCLUSION: This investigation reveals the molecular links between T2DM and AF with a focus on EAT. Targeting these pathways, especially EAT-related ones, may enable personalized treatments and improved outcomes.
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Fibrilação Atrial , Diabetes Mellitus Tipo 2 , Tecido Adiposo Epicárdico , Perfilação da Expressão Gênica , Pericárdio , Animais , Humanos , Masculino , Camundongos , Fibrilação Atrial/genética , Biologia Computacional/métodos , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/metabolismo , Tecido Adiposo Epicárdico/metabolismo , Perfilação da Expressão Gênica/métodos , Redes Reguladoras de Genes , Camundongos Endogâmicos C57BL , Pericárdio/metabolismo , Pericárdio/patologia , TranscriptomaRESUMO
Second harmonic generation plays a vital role in frequency conversion which mutually promotes the laser technology and allows the wavebands extension of new coherent source. The monolithic crystals are supposed to be a superior choice for harmonic generation due to long interaction distance, however, the phase-mismatch brought a sharp reduction in the conversion efficiency. Although birefringent phase-matching and quasi-phase-matching techniques are commonly utilized to fill the phase gap in monolithic crystals, these techniques are limited by the natural refractive index of crystal and the domain engineering, respectively. In recent years, subwavelength structures evolve as a flexible scheme to realize phase matching by engineering the geometry features of crystals. Here, structured nanogratings are designed and fabricated on a monolithic PMN-39PT (Pb(Mg1/3Nb2/3)O3-0.39PbTiO3) substrate, a novel ferroelectric crystal with promising optical prospect, for enhancing second harmonic generation, where birefringent or quasi phase-matching is hard to achieve. The nanograting-assisted second harmonic generation enhancement is observed which is not limited by the availability of thin crystalline films. Meanwhile, a boost in the second harmonic signal synchronously promotes the cascading third harmonic generation. This method may provide an alternative solution for enhanced harmonic generation on monolithic substrates and develop potential nonlinear optical materials for frequency conversion.
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BACKGROUND: Atrial Fibrillation (AF), a prevalent arrhythmic condition, is intricately associated with atrial fibrosis, a major pathological contributor. Central to the development of atrial fibrosis is myocardial inflammation. This study focuses on Atrial Natriuretic Peptide (ANP) and its role in mitigating atrial fibrosis, aiming to elucidate the specific mechanisms by which ANP exerts its effects, with an emphasis on fibroblast dynamics. METHODS AND RESULTS: The study involved forty Sprague-Dawley rats, divided into four groups: control, Angiotensin II (Ang II), Ang II + ANP, and ANP only. The administration of 1 µg/kg/min Ang II was given to Ang II and Ang II + ANP groups, while both Ang II + ANP and ANP groups received 0.1 µg/kg/min ANP intravenously for a duration of 14 days. Cardiac fibroblasts were used for in vitro validation of the proposed mechanisms. The study observed that rats in the Ang II and Ang II + ANP groups showed an increase in blood pressure and a decrease in body weight, more pronounced in the Ang II group. Diastolic dysfunction, a characteristic of the Ang II group, was alleviated by ANP. Additionally, ANP significantly reduced Ang II-induced atrial fibrosis, myofibroblast proliferation, collagen overexpression, macrophage infiltration, and the elevated expression of Interleukin 6 (IL-6) and Tenascin-C (TN-C). Transcriptomic sequencing indicated enhanced PI3K/Akt signaling in the Ang II group. Furthermore, in vitro studies showed that ANP, along with the PI3K inhibitor LY294002, effectively reduced PI3K/Akt pathway activation and the expression of TN-C, collagen-I, and collagen-III, which were induced by Ang II. CONCLUSIONS: The study demonstrates ANP's potential in inhibiting myocardial inflammation and reducing atrial fibrosis. Notably, ANP's effect in countering atrial fibrosis seems to be mediated through the suppression of the Ang II-induced PI3K/Akt-Tenascin-C signaling pathway. These insights enhance our understanding of AF pathogenesis and position ANP as a potential therapeutic agent for treating atrial fibrosis.
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Fibrilação Atrial , Fator Natriurético Atrial , Ratos , Animais , Ratos Sprague-Dawley , Fator Natriurético Atrial/farmacologia , Fator Natriurético Atrial/metabolismo , Proteínas Proto-Oncogênicas c-akt , Fosfatidilinositol 3-Quinases , Tenascina , Fibrilação Atrial/tratamento farmacológico , Angiotensina II/farmacologia , Inflamação/tratamento farmacológico , Colágeno , FibroseRESUMO
Birth defects are an important factor for the quality of newborn population. With the development of molecular genetic technology, an increasing number of genetic disorders leading to birth defects can now be detected. The lack of the knowledge for the basics and clinical applications of molecular genetic techniques have emerged as a shortcoming for primary care physicians who have formed the first tier prevention for birth defects. Currently, government has paid more attention to the above problems and formulated more training programs for primary obstetricians and gynecologists, e.g., "Prenatal Screening and Prenatal Diagnosis Post Training Program", "National Birth Defects Training Program", "National Primary Obstetrician Training Program". To some extent, such programs have met the urgent need for birth defect prevention in primary hospitals. But at the same time, some problems have also emerged. For instance, the knowledge for birth defects among primary obstetricians and gynecologists is poor, and there is lack of young personnel. This article has aimed to discuss the strategies to systematically improve the ability for preventing birth defects among primary care physicians by analyzing the obstacles and challenges for primary obstetricians and gynecologists in the era of molecular genetic testing.
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Ginecologia , Obstetrícia , Feminino , Gravidez , Recém-Nascido , Humanos , Ginecologista , Obstetra , Biologia MolecularRESUMO
Angiogenesis is essential for the growth and metastasis of several malignant tumors including colorectal cancer (CRC). The molecular mechanism underlying CRC angiogenesis has not been fully elucidated. Emerging evidence indicates that secreted microRNAs (miRNAs) may mediate the intercellular communication between tumor cells and neighboring endothelial cells to regulate tumor angiogenesis. In addition, exosomes have been shown to carry and deliver miRNAs to regulate angiogenesis. miRNA N-72 is a novel miRNA that plays a regulatory role in the EGF-induced migration of human amnion mesenchymal stem cells. However, the relation between miRNA N-72 and cancer remains unclear. We here found that CRC cells could secrete miRNA N-72. A high miRNA N-72 level was detected in the serum of CRC patients and the cultured CRC cells. Moreover, the CRC cell-secreted miRNA N-72 could promote the migration, tubulogenesis, and permeability of endothelial cells. In addition, the mouse xenograft model was used to verify the facilitating effects of miRNA N-72 on CRC growth, angiogenesis, and metastasis in vivo. Further mechanism analysis revealed that CRC cell-secreted miRNA N-72 could be delivered into endothelial cells via exosomes, which then inhibited cell junctions of endothelial cells by targeting CLDN18 and consequently promoted angiogenesis. Our findings reveal a novel mechanism of CRC angiogenesis and highlight the potential of secreted miRNA N-72 as a therapeutic target and a biomarker for CRC.
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An optical single-channel color image encryption scheme based on chaotic fingerprint phase mask and diffractive imaging is proposed. In this proposed encryption scheme, the fingerprint used to generate the random phase masks is served as a secret key directly. Additionally, the random phase masks generated by the fingerprint, chaotic Lozi map, and secure hash algorithm (SHA-256) are used only as interim variables. With the help of the chaotic fingerprint phase masks placed at different diffraction distances, the color image that is encoded into a grayscale pattern by the phase-truncation technique is encrypted into a noise-like diffraction pattern. For decryption, the color image can be retrieved from the noise-like diffraction pattern by using an iterative phase retrieval algorithm, fingerprint, and phase keys generated from the encryption process. Since the fingerprint key shared by the sender and authorized receiver is strongly linked with the user and does not need to be transmitted over the open network, the security of this proposed encryption scheme can be greatly improved. Additionally, the parameters of the chaotic Lozi map and Fresnel diffraction distances can also provide additional security to the proposed encryption scheme. Furthermore, compared with the encryption schemes based on digital holography, the implementation of this proposed encryption scheme is relatively simple. The numerical simulations and analysis verify the feasibility, security, and robustness of this proposed encryption scheme.
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In recent years, the thriving satellite laser communication industry has been severely hindered by the limitations of incompatible modulation formats and restricted Size Weight and Power (SWaP). A multi-modulation compatible method serving for free-space optical (FSO) communication has been proposed assisted by chirp-managed laser (CML). The corresponding demonstration system has been established for realizing free-switching between intensity (OOK) and phase modulation (RZ-DPSK). The feasibility and performance of system have been evaluated sufficiently when loading with 2.5 and 5 Gbps data streams, respectively. Additionally, a control-group system has been operated utilizing Mach-Zehnder modulator (MZM) for comparison between CML-based and MZM-based compatibility solutions. The OOK receiving sensitivities of CML-based system are -47.02 dBm@2.5 Gbps and -46.12 dBm@5 Gbps at BER of 1×10-3 which are 0.62 dB and 1.11 dB higher than that of MZM; the receiving sensitivities of RZ-DPSK are -50.12 dBm@2.5 Gbps and -47.03 dBm@5 Gbps which are 0.79 dB and 0.47 dB higher than that of MZM respectively. Meanwhile, CML-based transmitter abandoned the traditional modulator and its complicated supporting devices which can effectively contribute to the reduction of SWaP. The CML-based system has been proven to have the compatibility between intensity and phase modulation while also possesses a miniaturized design. It may provide fresh thinking to achieve a practical miniaturization system for satisfying the requirements of space optical network in future.
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A medium infrared 12× continuous zoom optical system serving for an acousto-optic tunable filter spectral camera has been designed covering a 25-300 mm zooming range. Instead of relying on past experience for roughly determining the initial optical structure, a simulation programming based on Gaussian principle has been composed via MATLAB to accurately calculate the initial designing parameters, which is confirmed to be extremely close to the optimized results using Zemax. The relative design results have been multidimensionally analyzed in detail, which offers fresh thinking for future zoom optical design covering broadband operation wave band.
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An asymmetric double-image encryption scheme based on chaotic random phase encoding (CRPE) is proposed. In this proposed encryption scheme, two grayscale images to be encrypted are first Fresnel transformed and combined into a complex image. Then, the amplitude and phase components are obtained by conducting phase-amplitude truncation on the complex image. Finally, the amplitude component is again Fresnel transformed and encrypted into a noise-like pattern by the CRPE in the Fresnel domain. Since the initial values and control parameters of the chaotic map can replace the random phase masks to serve as secret keys, the management and transmission of secret keys will become more convenient in the proposed encryption scheme. Furthermore, the Fresnel transform parameters and phase keys derived from the complex image's phase component can also act as secret keys during the decryption process. Numerical simulations have demonstrated the feasibility, security, and robustness of the proposed encryption scheme.
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A novel type of molybdenum disulfide (MoS2) nanoparticles (NPs) was chemically synthesized, which possessed curved features with three-dimensional (3D) freedom compared with planar two-dimensional (2D) materials. Due to the introduction of curved features, the synthesized NPs exhibited a strongly enhanced nonlinear refractive index (n2 â¼ 10-5 cm-2 W-1) and third-order susceptibility (χ(3) â¼ 10-7 esu), which were experimentally verified by the spatial self-phase modulation effect in the visible wavelength range. Both the nonlinear parameters were two orders of magnitude higher than their planar MoS2 nanostructure counterparts. In addition, the relative change of the effective nonlinear refractive index Δn2/n2 was found to be distinctly dependent on the intensity of the applied electromagnetic field. Moreover, an all-optical modulation was experimentally realized based on the spatial cross-phase modulation effect. Our results demonstrate planar MoS2 materials with 3D features as potential candidates for next generation all-optical applications and open a substantial approach for the design of efficient nanomaterials with favorable optical nonlinearity.
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BACKGROUND: Gastrointestinal dysfunction is one of the complications after stroke. If it is not treated in time, it will affect the rehabilitation process after stroke and reduce the quality of life of patients. In this study, we conducted a systematic review and meta-analysis of the reports on the treatment of gastrointestinal dysfunction after stroke with traditional Chinese medicine (TCM) in recent years to provide evidence for clinical diagnosis and treatment. METHODS: The clinical randomized controlled trials (RCTs) published in Chinese Biomedical Literature Database (CBM), China National Knowledge Infrastructure (CNKI), Medline, and Web of Science databases from January 2010 to August 2021 were searched. After screening the qualified literatures, literature quality evaluation was performed. The software Stata 16.0 was used to analyze and compare the outcome indicators of TCM and conventional western medicine treatment, and the utility of TCM in the treatment of gastrointestinal disorders after stroke was comprehensively evaluated. RESULTS: A total of 16 studies were finally selected including a total of 1,589. Meta-analysis showed that TCM treatment of gastrointestinal disorders after stroke was more effective than conventional western medicine treatment [odds ratio (OR) =3.94; 95% confidence interval (CI): 2.63 to 5.89; P=0.000]. It was also shown that TCM can reduce the recovery time of bowel sounds and is superior to conventional western medicine [standard mean difference (SMD) =-1.92; 95% CI: -2.51 to -1.34; P=0.000]; reduce defecation and flatulence recovery time (SMD =-2.51; 95% CI: -3.41 to -1.61; P=0.000); increase gastrin level (SMD =0.80; 95% CI: 0.35 to 1.25; P=0.001); increase motilin level (SMD =2.27; 95% CI: 1.55 to 3.00; P=0.000); increase serum albumin level (SMD =0.72; 95% CI: 0.54 to 0.90; P=0.000); increase transferrin levels (SMD =1.11; 95% CI: 0.86 to 1.36; P=0.000); and it can increase serum prealbumin levels (SMD =1.50; 95% CI: 0.78 to 2.22; P=0.000). DISCUSSION: The use of TCM in the treatment of gastrointestinal dysfunction after stroke can effectively reduce symptoms, shorten the recovery time of bowel sounds, the first defecation and flatulence time, promote gastric motility and gastrointestinal hormone secretion, and improve the nutritional status of patients.
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Gastroenteropatias , Acidente Vascular Cerebral , Gastroenteropatias/etiologia , Humanos , Medicina Tradicional Chinesa , Qualidade de Vida , Ensaios Clínicos Controlados Aleatórios como Assunto , Acidente Vascular Cerebral/complicaçõesRESUMO
We observed the generation of phase-matching conical second harmonic generation (SHG) inside KDP crystal under anomalous dispersion condition, which is attributed to complete phase-matching assisted by fundamental wave (FW) and scattering wave. The double-ring pattern of the conical SHG implies that elastic scattering can stimulate two different polarization states of scattering wave. Furthermore, SH ring of KDP displays inhomogeneous intensity around the ring, which has relevance to effective nonlinear coefficient, and it can be used to investigate crystal symmetry.
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Clones of the WWE1 (Waste Water of Evry 1) candidate division were retrieved during the exploration of the bacterial diversity of an anaerobic mesophilic (35 ± 0.5°C) digester. In order to investigate the metabolic function of WWE1 members, a 16S rRNA gene -based stable isotope probing (SIP) method was used. Eighty-seven percent of 16S r rRNA gene sequences affiliated to WWE1 candidate division were retrieved in a clone library obtained after polymerase chain reaction (PCR) amplification of enriched DNA fraction from anaerobic municipal solid waste samples incubated with (13) C-cellulose, at the end of the incubation (day 63) using a Pla46F-1390R primer pair. The design of a specific WWE1 probe associated with the fluorescence in situ hybridization (FISH) technique corroborated the abundant representation of WWE1 members in our (13) C-cellulose incubations. Secondary ion mass spectrometry-in situ hybridization (SIMSISH) using an iodine-labeled oligonucleotide probe combined with high-resolution nanometer-scale SIMS (NanoSIMS) observation confirmed the isotopic enrichment of members of WWE1 candidate division. The (13) C apparent isotopic composition of hybridized WWE1 cells reached the value of about 40% early during the cellulose degradation process, suggesting that these bacteria play a role either in an extracellular cellulose hydrolysis process and/or in the uptake fermentation products.
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Bactérias/isolamento & purificação , Bactérias/metabolismo , Celulose/metabolismo , Águas Residuárias/microbiologia , Anaerobiose , Bactérias/genética , Biota , Isótopos de Carbono/metabolismo , Hibridização in Situ Fluorescente , Marcação por IsótopoRESUMO
We collected samples of anaerobic landfill leachate from municipal solid waste landfill (Vert-le-Grand, France) and constructed 16S rRNA clone libraries using primers targeting Planctomycetes and relatives (Pla46F and 1390R). Analyses of 16S rRNA gene sequences resulted in the abundant representation of WWE2-related Lentisphaerae, members of the phylum Lentisphaerae, in the clone library (98% of the retrieved sequences). Although the sequences that are phylogenetically affiliated with the cultured isolate Victivallis vadensis were identified (WWE2 subgroup II), the majority of the sequences were affiliated with an uncultured Lentisphaerae lineage (WWE2 subgroup I). We designed oligonucleotides probes targeting the specific 16S rRNA gene regions of those 2 subgroups. Fluorescence in situ hybridization confirmed the abundance of the uncultivated WWE2 subgroup I in our leachate samples.
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Bactérias/classificação , Bactérias/isolamento & purificação , Resíduos de Alimentos , Genes de RNAr , Hibridização in Situ Fluorescente , Anaerobiose , Bactérias/genética , Biomassa , DNA Bacteriano/análise , DNA Ribossômico/análise , França , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Eliminação de Resíduos , Análise de Sequência de DNA , Microbiologia do Solo , Poluentes do Solo , Gerenciamento de Resíduos , Poluentes Químicos da ÁguaRESUMO
The microbial consortium involved in anaerobic digestion has not yet been precisely characterized and this process remains a 'black box' with limited efficiency. In this study, seven anaerobic sludge digesters were selected based on technology, type of sludge, process and water quality. The prokaryotic community of these digesters was examined by constructing and analysing a total of 9890 16S rRNA gene clones. Libraries were constructed using primers specific for the Bacteria and Archaea domains for each digester, respectively. After phylogenetic affiliation, the libraries were compared using statistical tools to determine the similarities or differences among the seven digesters. Results show that the prokaryotic community of an anaerobic digester is composed of phylotypes commonly found in all anaerobic digesters sampled and also of specific phylotypes. The Archaea community is represented by an equilibrium among a restricted number of operational taxonomic units (OTUs). These OTUs are affiliated with Methanosarcinales, Methanomicrobiales and Arc I phylogenetic groups. Statistical analysis revealed that the Bacteria community can be described as a three component model: one-third making up a core group of phylotypes common to most of the digesters, one-third are phylotypes shared among a few digesters and another one-third are specific phylotypes. The core group is composed of only six OTUs affiliated with Chloroflexi, Betaproteobacteria, Bacteroidetes and Synergistetes. Its role in anaerobic degradation appears critical to investigate. This comparison of anaerobic digester populations is a first step towards a future understanding of the relationship among biodiversity, operating conditions and digester efficiency.
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Archaea/classificação , Archaea/isolamento & purificação , Bactérias/classificação , Bactérias/isolamento & purificação , Biodiversidade , Esgotos/microbiologia , Anaerobiose , Archaea/metabolismo , Bactérias/metabolismo , Análise por Conglomerados , DNA Arqueal/química , DNA Arqueal/genética , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Homologia de Sequência do Ácido NucleicoRESUMO
DNA-SIP (stable isotope probing) was conducted on anaerobic municipal solid waste samples incubated with (13)C-cellulose, (13)C-glucose and (13)C-acetate under mesophilic conditions. A total of 567 full-length bacterial and 448 1100-bp-length archaeal 16S rRNA gene sequences were analysed. In the clone libraries derived from 'heavy' DNA fractions, the most abundant sequences were affiliated with the phyla Firmicutes, Bacteroidetes, the gamma-subclass of Proteobacteria and methanogenic orders Methanomicrobiales and Methanosarcinales. Sequences related to the genus Acetivibrio (phylum Firmicutes) were recovered only in the 'heavy' DNA fraction derived from the (13)C-cellulose incubation. An oligonucleotide probe (UCL284) targeting specifically Acetivibrio was designed and used for fluorescent in situ hybridization (FISH) experiments. Interestingly, hybridization of the probe was detected in microorganisms aggregated around cellulose fibres, strengthening the conclusion that these microorganisms were major cellulose degraders. Sequences related to genus Clostridium (phylum Firmicutes) and to the family Porphyromonadaceae (phylum Bacteroidetes) were retrieved in large numbers from the 'heavy' DNA library of (13)C-Glucose incubation, suggesting their involvement in saccharide fermentation. Design and hybridization of specific FISH-probes confirmed the abundant representation of Clostridium (CLO401, CLO1248) and Porphyromonadaceae (BAC1040), which were mostly observed in the planktonic phase. Surprisingly, in the (13)C-acetate experiment, the 'heavy' DNA archaeal library was dominated by sequences related to the strictly hydrogenotrophic methanogenic genus Methanoculleus. One single operational taxonomic unit containing 70 sequences, affiliated to the gamma-subclass of Proteobacteria, was retrieved in the corresponding bacterial library. FISH observations with a newly designed specific probe (UGA64) confirmed the dominance of this bacterial group. Our results show that combination of DNA-SIP and FISH applied with a series of functionally connected substrates can shed light on the networks of uncultured microbes catalysing the methanization of the most abundant chemical renewable energy source on Earth.
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Archaea/classificação , Archaea/metabolismo , Bactérias/classificação , Bactérias/metabolismo , Biodiversidade , Celulose/metabolismo , Microbiologia Ambiental , Archaea/genética , Archaea/isolamento & purificação , Bactérias/genética , Bactérias/isolamento & purificação , Isótopos de Carbono/metabolismo , DNA Arqueal/química , DNA Arqueal/genética , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Genes de RNAr , Hibridização in Situ Fluorescente , Dados de Sequência Molecular , Filogenia , RNA Arqueal/genética , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Homologia de Sequência do Ácido NucleicoRESUMO
Identifying the function of uncultured microbes in their environments today remains one of the main challenges for microbial ecologists. In this article, we describe a new method allowing simultaneous analysis of microbial identity and function. This method is based on the visualization of oligonucleotide probe-conferred hybridization signal in single microbial cells and isotopic measurement using high-resolution ion microprobe (NanoSIMS). In order to characterize the potential of the method, an oligonucleotide containing iodized cytidine was hybridized on fixed cells of Escherichia coli cultured on media containing different levels of 13C or 15N. Iodine signals could clearly be localized on targeted cells and the isotopic enrichment could be monitored at the single-cell level. The applicability of this new technique to the study of in situ ecophysiology of uncultured microorganisms within complex microbial communities is illustrated.
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Técnicas de Tipagem Bacteriana/métodos , Microbiologia Ambiental , Hibridização In Situ , Sondas de Oligonucleotídeos/metabolismo , Isótopos de Carbono , Técnicas de Laboratório Clínico , Técnicas Microbiológicas , Sondas de Oligonucleotídeos/genéticaRESUMO
Archaeal microbial communities present in municipal solid waste landfill leachates were characterized using a 16S rDNA approach. Phylogenetic affiliations of 239 partial length 16S rDNA sequences were determined. Sequences belonging to the order Methanosarcinales were dominant in the clone library and 65% of the clones belonged to the strictly acetoclastic methanogenic family Methanosaetaceae. Sequences affiliated to the metabolically versatile family Methanosarcinaceae represented 18% of the retrieved sequences. Members of the hydrogenotrophic order Methanomicrobiales were also recovered in limited numbers, especially sequences affiliated to the genera Methanoculleus and Methanofollis. Eleven euryarchaeal and thirteen crenarchaeal sequences (i.e. 10%) were distantly related to any hitherto cultivated microorganisms, showing that archaeal diversity within the investigated samples was limited. Lab-scale incubations were performed with leachates mixed with several methanogenic precursors (acetate, hydrogen, formate, methanol, methylamine). Microbial populations were followed using group specific 16S rRNA targeted fluorescent oligonucleotidic probes. During the incubations with acetate, acetoclastic methanogenesis was rapidly induced and led to the dominance of archaea hybridizing with probe MS1414 which indicates their affiliation to the family Methanosarcinaceae. Hydrogen and formate addition induced an important acetate synthesis resulting from the onset of homoacetogenic metabolism. In these incubations, species belonging to the family Methanosarcinaceae (hybridizing with probe MS1414) and the order Methanomicrobiales (hybridizing with probe EURY496) were dominant. Homoacetogenesis was also recorded for incubations with methanol and methylamines. In the methanol experiment, acetoclastic methanogenesis took place and archaea hybridizing with probe MS821 (specific for Methanosarcina spp.) were observed to be the dominant population. These results confirm that acetoclastic methanogenesis performed by the members of the order Methanosarcinales is predominant over the hydrogenotrophic and methylotrophic pathways in landfill leachates.