RESUMO
Microbial carbon utilization efficiency (CUE) is a crucial indicator for evaluating the efficiency of soil carbon sequestration and transformation, which is applied to quantify the proportion of soil carbon extracted by microbes for anabolism (growth) and catabolism (respiration). Previous studies have shown that the degradation of Moso bamboo forests (Phyllostachys edulis) destroyed the aboveground bamboo structure, reduced vegetation carbon storage, and weakened ecosystem carbon sequestration capacity. Interestingly, soil organic carbon stocks are gradually increasing. However, the mechanism by which degradation-induced changes in soil and vegetation characteristics affect microbial CUE and drive soil carbon sequestration remains unclear. Here we selected four stands with the same origin but different degradation years (intensive management, CK; 2 years' degradation, DM1; 6 years' degradation, DM2; and 10 years' degradation, DM3) based on the local management profiles. The principle of space-for-time substitution was used to investigate the changes in microbial CUE along a degradation time and to further identify the controlling biotic and abiotic factors. Our finding showed that microbial CUE increased by 12.27 %, 31.01 %, and 55.95 %, respectively, compared with CK; whereas microbial biomass turnover time decreased from 23.99 ± 1.11 to 17.16 ± 1.20 days. Promoting microbial growth was the main pathway to enhance microbial CUE. Massive inputs of vegetative carbon replenished soil carbon substrate content, and altered microbial communities and life history strategy, which in turn promoted microbial growth and increased microbial CUE. These findings provide theoretical support for the interactions between carbon dynamics and microbial physiology in degraded bamboo forests, and reinforce the importance of vegetation and microbial properties and soil carbon substrates in predicting microbial CUE.
RESUMO
Moso bamboo (Phyllostachys heterocycla cv. Pubescens) is well known for its high capacity to sequester atmospheric carbon, which has a unique role to play in combating global warming. Many Moso bamboo forests are gradually degrading due to rising labor costs and falling prices for bamboo timber. However, the mechanisms of carbon sequestration of Moso bamboo forest ecosystems in response to degradation are unclear. In this study, a space-for-time substitution approach was used to select Moso bamboo forest plots with the same origin and similar stand types, but different years of degradation, and four degradation sequences, continuous management (CK), 2 years of degradation (D-I), 6 years of degradation (D-II) and 10 years of degradation (D-III). A total of 16 survey sample plots were established based on the local management history files. After a 12-month monitoring, the response characteristics of soil greenhouse gases (GHG) emissions, vegetation, and soil organic carbon sequestration in different degradation sequences were evaluated to reveal the differences in the ecosystem carbon sequestration. The results indicated that under D-I, D-II, and D-III, the global warming potential (GWP) of soil GHG emissions decreased by 10.84 %, 17.75 %, and 31.02 %, while soil organic carbon (SOC) sequestration increased by 2.82 %, 18.11 %, and 4.68 %, and vegetation carbon sequestration decreased by 17.30 %, 33.49 %, and 44.76 %, respectively. In conclusion, compared to CK, the ecosystem carbon sequestration was reduced by 13.79 %, 22.42 %, and 30.31 %, respectively. This suggests that degradation reduces soil GHG emissions but weakens the ecosystem carbon sequestration capability. Therefore, in the background of global warming and the strategic goal of carbon neutrality, restorative management of degraded Moso bamboo forests is critically needed to improve the carbon sequestration potential of the ecosystem.
Assuntos
Ecossistema , Gases de Efeito Estufa , Sequestro de Carbono , Gases de Efeito Estufa/metabolismo , Carbono/análise , Solo , Poaceae/metabolismo , ChinaRESUMO
A novel and sensitive assay for aflatoxin B1 (AFB1) detection has been developed by using bio-bar code assay (BCA). The method that relies on polyclonal antibodies encoded with DNA modified gold nanoparticle (NP) and monoclonal antibodies modified magnetic microparticle (MMP), and subsequent detection of amplified target in the form of bio-bar code using a fluorescent quantitative polymerase chain reaction (FQ-PCR) detection method. First, NP probes encoded with DNA that was unique to AFB1, MMP probes with monoclonal antibodies that bind AFB1 specifically were prepared. Then, the MMP-AFB1-NP sandwich compounds were acquired, dehybridization of the oligonucleotides on the nanoparticle surface allows the determination of the presence of AFB1 by identifying the oligonucleotide sequence released from the NP through FQ-PCR detection. The bio-bar code techniques system for detecting AFB1 was established, and the sensitivity limit was about 10-8 ng/mL, comparable ELISA assays for detecting the same target, it showed that we can detect AFB1 at low attomolar levels with the bio-bar-code amplification approach. This is also the first demonstration of a bio-bar code type assay for the detection of AFB1 in Chinese herbs.
Assuntos
Aflatoxina B1/análise , Medicamentos de Ervas Chinesas/análise , Processamento Eletrônico de Dados/métodos , Aflatoxina B1/genética , Contaminação de Medicamentos/estatística & dados numéricos , Processamento Eletrônico de Dados/instrumentação , Ensaio de Imunoadsorção Enzimática , Ouro/química , Nanopartículas Metálicas/química , Reação em Cadeia da PolimeraseRESUMO
A long-lasting recombinant human serum albumin-interferon-alpha2b fusion protein (rHSA/IFNalpha2b) was prepared and its structure and biological activities were studied. rHSA/IFNalpha2b was expressed in methylotrophic yeast Pichia pastoris with HSA's natural signal peptide and purified by dye affinity chromatography, hydrophobic interaction chromatography, ion exchange chromatography and Sephadex G25. Purity of the prepared rHSA/IFNalpha2b was greater than 97% analyzed by non-reduced SDS-PAGE and RP-HPLC. Structure and biological activities of the prepared rHSA/IFNalpha2b were characterized by physical, chemical and biological methods. Its pI was 5.3 and showed a single band on IEF gel. Molecular weight determined by MALDI-TOF was 86004.3+/-29.2. Amino-terminal and carboxyl-terminal amino acid sequences were identical to predicted sequence. Its specific activity in vitro was 6.3+/-0.8x10(5) IU/mg fusion protein, retaining about 1.4% of that of unmodified rIFNalpha on a molar basis. After administered in monkeys, significant increases of 2',5'-oligoadenylate synthetase activity relative to IFN-alpha were maintained for 14 days in serum and the rHSA/IFNalpha2b showed more potent biological activity than IFN-alpha on a molar basis. Therefore, markedly improved in vivo biological activity of rHSA/IFNalpha2b could exhibit more potent antiviral activity than IFNalpha2b in future clinical trials.
Assuntos
Interferon-alfa/química , Pichia/metabolismo , Proteínas Recombinantes de Fusão/química , Albumina Sérica/química , Sequência de Aminoácidos , Antivirais/farmacologia , Química Farmacêutica/métodos , Cromatografia , Cromatografia por Troca Iônica/métodos , Humanos , Interferon alfa-2 , Focalização Isoelétrica , Dados de Sequência Molecular , Peso Molecular , Sinais Direcionadores de Proteínas , Proteínas Recombinantes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
To reduce the serum clearance of interferon alpha2b, a chimeric gene encoding an human serum albumin(HSA)--human interferon alpha2b(IFNalpha2b) fusion protein was overexpressed in Pichia pastoris. After fermentation in a 5L bioreactor, the fusion protein, capable of cross-reacting with anti-IFN alpha and anti-HSA antibody, was purified from the culture of the recombinant yeast by ultrafiltration, blue Sepharose affinity, phenyl hydrophobic interaction and Q ion exchange chromatography. Its IFNa2b moiety exhibits antiviral activity similar to that of recombinant human IFNa2b. In Cynomolgus monkeys model, The fusion protein was detectable in plasma, even 336h after a single does of 90 microg/kg injection intravenously or subcutaneously. The elimination phase half-life of the fusion protein was 101h after intravenous injection and 68.2h after subcutaneous injection. Its Subcutaneous bioavailability was 67.9%. The enhanced pharmacokinetics of interferon a2b fused to human serum albumin suggest its promissing application in clinic medicine.
Assuntos
Interferon-alfa/genética , Proteínas Recombinantes de Fusão/farmacocinética , Albumina Sérica/genética , Animais , Reatores Biológicos/microbiologia , Fermentação , Humanos , Interferon alfa-2 , Interferon-alfa/biossíntese , Macaca fascicularis , Pichia/genética , Pichia/metabolismo , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes , Albumina Sérica/biossínteseRESUMO
We demonstrated previously that hypoxia activated CRF and CRF mRNA in PVN, and CRF receptor 1 (CRFR1) mRNA in rat pituitary. The aim of the study is to test whether the hypoxia-activated CRF and CRF mRNA is associated with triggering CRFR1. Rats were exposed to hypobaric hypoxia at altitude of 2 and 5 km. CRF and CRF mRNA were assayed by immunostaining and in situ hybridization. CRFR1 mRNA was assayed by RT-PCR. Results showed that 5 km continual hypoxia increased CRF and CRF mRNA in PVN, CRFR1 mRNA in pituitary, and plasma corticosterone. The hypoxia-increased CRF, CRF mRNA, CRFR1 mRNA, and corticosterone were blocked by CRFR1 antagonist (CP-154,526), suggesting that CRFR1 in PVN and pituitary are responsible for the hypoxia-increased CRF and CRF mRNA in PVN.
Assuntos
Hormônio Adrenocorticotrópico/fisiologia , Regulação da Expressão Gênica , Núcleo Hipotalâmico Paraventricular/fisiologia , RNA Mensageiro/genética , Receptores de Hormônio Liberador da Corticotropina/fisiologia , Hormônio Adrenocorticotrópico/genética , Animais , Hipóxia Celular , Corticosterona/sangue , Corticosterona/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Masculino , Hipófise/fisiologia , Pirimidinas/farmacologia , Pirróis/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de Hormônio Liberador da Corticotropina/antagonistas & inibidores , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVES: The aim of this study was to investigate the response of the growth hormone (GH) in rat anterior pituitary to intermittent hypoxia (IH) and its modulation by hypothalamic somatostatin (SS). SETTING AND DESIGN: To observe the hypoxic response, rats were exposed to simulated altitude hypoxia (2 km or 5 km) in a hypobaric chamber for various days (4 h/d); to clarify SS-involvement, rats were pretreated with SS antagonist (cysteamine, CSH, 200 mg/kg/d, s.c.) then exposed to IH (5 km) for 2d. The GH mRNA and immunostaining GH in pituitary as well as immunostaining SS in median eminence (ME) of hypothalamus were assayed by RT-PCR and immuno-histochemistry, respectively. RESULTS: IH of 5 km altitude (IH5) markedly suppressed the body weight gain (BWG) of rats from 1d to 10d, and it was returned to control level henceforth, while no significant influence was showed in the group of IH of 2 km altitude (IH2). IH5 for 2 and 5d significantly decreased GH mRNA expression in the pituitary. The pituitary immunostaining GH was remarkably increased in groups of IH2 for 5, 10, and 15 d, and in groups of IH5 for 2, 5, and 10d. Immunostaining SS in ME was significantly reduced in group of IH2 for 5d, and in groups of IH5 for 2d and 5d. Pretreatments (s.c.) with SS antagonist (CSH) significantly reversed IH5-caused increase of immunostaining GH and reduction of mRNA levels in pituitary. CONCLUSIONS: IH may cause a short-term and recoverable suppression of GH release, and reduce GH mRNA expression in anterior pituitary, which may depend on the intensity and duration of the hypoxia. This suppression may be due to a modulation of hypoxia-activated SS.