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1.
Int J Biol Macromol ; 273(Pt 2): 133135, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38876231

ABSTRACT

Largemouth bass (Micropterus salmoides) has emerged as a significant economic fish species, with a rise in Aeromonas veronii infections in farming. However, research on adjuvants for vaccines against A. veronii in largemouth bass remains scarce. In present study, recombinant largemouth bass IL-1ß (LbIL-1ß) was expressed to explore its adjuvant effect on the A. veronii inactivated vaccine. Following vaccination with recombinant LbIL-1ß (rLbIL-1ß) and the inactivated A. veronii, higher serum SOD levels and lysozyme activities were observed in largemouth bass from inactivated A. veronii + rLbIL-1ß vaccinated group. Furthermore, it was discovered that rLbIL-1ß was able to boost the serum-specific antibody levels induced by the inactivated A. veronii. The qRT-PCR analysis revealed that rLbIL-1ß also enhanced the expression of IgM, CD4, and MHC II in largemouth bass triggered by the inactivated A. veronii. After challenged with live A. veronii, the outcomes demonstrated that the relative percentage survival (RPS) for largemouth bass resulting from the inactivated A. veronii in combination with rLbIL-1ß was 76.67 %, surpassing the RPS of 60 % in the inactivated A. veronii group. Collectively, these findings indicate that rLbIL-1ß enhances the protective effect of the A. veronii inactivated vaccine on largemouth bass, showcasing potential as an adjuvant for further development.


Subject(s)
Adjuvants, Immunologic , Aeromonas veronii , Bacterial Vaccines , Bass , Fish Diseases , Interleukin-1beta , Vaccines, Inactivated , Animals , Bass/immunology , Bass/microbiology , Adjuvants, Immunologic/pharmacology , Aeromonas veronii/immunology , Vaccines, Inactivated/immunology , Fish Diseases/immunology , Fish Diseases/prevention & control , Fish Diseases/microbiology , Bacterial Vaccines/immunology , Gram-Negative Bacterial Infections/immunology , Gram-Negative Bacterial Infections/veterinary , Gram-Negative Bacterial Infections/prevention & control , Vaccination
2.
Sci Rep ; 13(1): 18498, 2023 10 28.
Article in English | MEDLINE | ID: mdl-37898687

ABSTRACT

Laryngeal squamous cell carcinoma (LSCC) is a common tumor type. High recurrence rates remain an important factor affecting the survival and quality of life of advanced LSCC patients. We aimed to build a new nomogram and a random survival forest model using machine learning to predict the risk of LSCC progress. The study included 671 patients with AJCC stages III-IV LSCC. To develop a prognostic model, Cox regression analyses were used to assess the relationship between clinic-pathologic factors and disease-free survival (DFS). RSF analysis was also used to predict the DFS of LSCC patients. The ROC curve revealed that the Cox model exhibited good sensitivity and specificity in predicting DFS in the training and validation cohorts (1 year, validation AUC = 0.679, training AUC = 0.693; 3 years, validation AUC = 0.716, training AUC = 0.655; 5 years, validation AUC = 0.717, training AUC = 0.659). Random survival forest analysis showed that N stage, clinical stage, and postoperative chemoradiotherapy were prognostically significant variables associated with survival. The random forest model exhibited better prediction ability than the Cox regression model in the training cohort; however, the two models showed similar prediction ability in the validation cohort.


Subject(s)
Carcinoma, Squamous Cell , Head and Neck Neoplasms , Humans , Squamous Cell Carcinoma of Head and Neck , Proportional Hazards Models , Carcinoma, Squamous Cell/pathology , Quality of Life , Prognosis , Machine Learning
3.
Antioxidants (Basel) ; 12(4)2023 Apr 12.
Article in English | MEDLINE | ID: mdl-37107289

ABSTRACT

Polysaccharide decolorization has a major effect on polysaccharide function. In the present study, the decolorization of Rehmannia glutinosa polysaccharides (RGP) is optimized using two methods-the AB-8 macroporous resin (RGP-1) method and the H2O2 (RGP-2) method. The optimal decolorization parameters for the AB-8 macroporous resin method were as follows: temperature, 50 °C; macroporous resin addition, 8.4%; decolorization duration, 64 min; and pH, 5. Under these conditions, the overall score was 65.29 ± 3.4%. The optimal decolorization conditions for the H2O2 method were as follows: temperature, 51 °C; H2O2 addition, 9.5%; decolorization duration, 2 h; and pH, 8.6. Under these conditions, the overall score was 79.29 ± 4.8%. Two pure polysaccharides (RGP-1-A and RGP-2-A) were isolated from RGP-1 and RGP-2. Subsequently, their antioxidant and anti-inflammatory effects and mechanisms were evaluated. RGP treatment activated the Nrf2/Keap1 pathway and significantly increased the activity of antioxidant enzymes (p < 0.05). It also inhibited the expression of pro-inflammatory factors and suppressed the TLR4/NF-κB pathway (p < 0.05). RGP-1-A had a significantly better protective effect than RGP-2-A, likely owing to the sulfate and uronic groups it contains. Together, the findings indicate that RGP can act as a natural agent for the prevention of oxidation and inflammation-related diseases.

4.
Foods ; 11(21)2022 Oct 31.
Article in English | MEDLINE | ID: mdl-36360063

ABSTRACT

Polysaccharide decolorization is a key determinant of polysaccharide structure. In this study, two purified Rehmannia glutinosa polysaccharides, RGP-1-A and RGP-2-A, were obtained after decolorization using the AB-8 macroporous resin and H2O2, respectively. RGP-1-A (molecular weight (Mw) = 18,964 Da) and RGP-2-A (Mw = 3305 Da) were acidic and neutral heteropolysaccharides, respectively, and were both polycrystalline in structure. FTIR analysis revealed that RGP-1-A was a sulfate polysaccharide, while RGP-2-A had no sulfate group. Experiments on IPEC-1 cells showed that RGPs alleviated oxidative stress by regulating the Nrf2/Keap1 pathway. These findings were confirmed by the upregulation of Nrf2, NQO1, and HO-1; the subsequent increase in the levels of antioxidant indicators (SOD, LDH, CAT, and MDA); and the restoration of mitochondrial membrane potential. Overall, the antioxidant capacity of RGP-1-A was significantly higher than that of RGP-2-A. These results suggest that RGPs may be a potential natural antioxidant and could be developed into functional foods.

5.
Technol Cancer Res Treat ; 21: 15330338221107710, 2022.
Article in English | MEDLINE | ID: mdl-35815926

ABSTRACT

Background: Head and neck squamous cell carcinoma (HNSCC) was the seventh most common cancer worldwide in 2018. Lymphatic metastasis (LM) is closely related to HNSCC prognosis and recurrence. However, the underlying mechanism of LM remains unclear. Therefore, this study aimed to identify the key genes in the LM of HNSCC. Methods: We used The Cancer Genome Atlas (TCGA) to identify differentially expressed genes (DEGs) between LM and non-LM cases. A random forest model, the Search Tool for the Retrieval of Interacting Genes, Cytoscape, and cytoHubba were used to identify hub genes among DEGs, including KRT20 (Cytokeratins 20). We analyzed the survival of KRT20 in TCGA, and we overexpressed KRT20 in HNSCC cell lines to investigate its effects on migration and invasion. We also correlated the expression of KRT20 in HNSCC tissue microarrays with survival and clinicopathological features. Results: We identified 243 DEGs-143 upregulated genes and 100 downregulated genes. Further analysis revealed that KRT20 is a potential key gene associated with LM and overall survival rates among patients with HNSCC. Overexpression of KRT20 increased the migration and invasion ability of HNSCC cell lines Tu686 and FD-LSC-1. Tissue microarray studies demonstrated an overexpression of KRT20 among N1+ patients (including N1-N3 patients). Survival analysis results and the clinicopathological features of HNSCC tissue microarrays were consistent with our analysis of TCGA. Thus, a high KRT20 expression level might suggest an adverse HNSCC prognosis. Our gene set enrichment analysis showed that KRT20 participates in many metabolic pathways, including those related to tumorigenesis and cancer development. Conclusions: We propose that KRT20 may be a key gene in HNSCC with LM.


Subject(s)
Head and Neck Neoplasms , Biomarkers, Tumor/genetics , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms/genetics , Humans , Keratin-20/genetics , Lymphatic Metastasis , Prognosis , Squamous Cell Carcinoma of Head and Neck/genetics
6.
Nat Prod Res ; 36(9): 2239-2245, 2022 May.
Article in English | MEDLINE | ID: mdl-32993365

ABSTRACT

Four p-coumaroylated sesquiterpenoids (1-4), including three new ones (1, 2, and 4), were isolated from the whole plants of Pilea cavaleriei. Their structures were established on the basis of comprehensive spectroscopic analysis and comparison with data reported in the literature. Compounds 1-4 were evaluated for their antimycobacterial activities, and 2 and 4 were active against Mycobacterium tuberculosis H37Rv with the MIC values of 19.6 ± 0.19 and 17.4 ± 0.12 µg/mL, respectively.


Subject(s)
Drugs, Chinese Herbal , Mycobacterium tuberculosis , Sesquiterpenes , Urticaceae , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Drugs, Chinese Herbal/chemistry , Microbial Sensitivity Tests , Sesquiterpenes/chemistry , Urticaceae/chemistry
7.
Environ Sci Pollut Res Int ; 27(14): 17085-17092, 2020 May.
Article in English | MEDLINE | ID: mdl-32146663

ABSTRACT

The wetland plants are very sensitive to hydrological regimes. In this study, the individual sizes of a widely distributed species (i.e., Deyeuxia angustifolia) at three typical marshes with different water table depths (i.e., wet meadow (WM) marsh; seasonal inundated (SI) marsh; perennial inundated (PI) marsh) were investigated in the Sanjiang Plain of Northeast China. Concurrently, three primary point pattern processes (homogeneous Poisson (HP) process, homogeneous Thomas (HT) process, and inhomogeneous Thomas (IT) process) were used to model spatial patterns in the distribution at 0-50 cm scale for this tillering-cloning species. The plant height, diameter at breast height (d.b.h), internode number, branches number, and individual aboveground biomass of D. angustifolia decreased sharply with rising water level; however, its density and coverage increased first and then decreased as water level increases. The distribution of D. angustifolia totally diverged from the complete spatial randomness (CSR) model (i.e., HP process) suggesting strong aggregation at 0-50 cm scale in all marshes, and aggregated intensity enhanced with increasing water level. Interestingly, the spatial distribution of D. angustifolia fits better with the nested double-cluster model (i.e., IT process) at all scales in WM and SI marshes, indicating that there is a series of clustered patterns under the slight flood stress. However, the spatial pattern fits well with the Poisson cluster model (i.e., HT process) at all scales in PI marsh, implying the small-scale clustering disappeared with the intensification of flooding stress. Our results highlight that the D. angustifolia population could adapt to flooding stress in a certain degree via individual miniaturization strategies and multi-aggregation mechanisms in the freshwater wetlands.


Subject(s)
Water , Wetlands , Biomass , China , Fresh Water , Poaceae
8.
Environ Sci Pollut Res Int ; 26(20): 20469-20474, 2019 Jul.
Article in English | MEDLINE | ID: mdl-31102213

ABSTRACT

The expansion of poisonous plants can change vegetation community structures and affect grassland ecosystem service values. Stellera chamaejasme is one of the most important poisonous plants and has rapidly expanded in the arid areas of Northwest China in recent decades. The objective of this study was to elucidate the expansion process and model of an S. chamaejasme population. Therefore, we classified the S. chamaejasme population into five classes based on coverage: 31-40%, 41-50%, 51-60%, 61-70% and 71-80%. We investigated the spatial distribution patterns and the size compositions of S. chamaejasme under different coverages. The results show that the spatial distribution pattern of S. chamaejasme under low coverage (31-40%) at all study scales (0-100 cm) was random; the spatial distribution pattern translated to a clumped distribution from a random distribution at some scales, and the clumped distributions gradually became obvious, with coverage increasing from 41-50% to 61-70%; the spatial distribution tended to be random at all study scales when coverage was increased further (71-80%). However, the spatial distribution patterns were closely related to the size composition of the S. chamaejasme population. In particular, the quantity of older individuals had a significant impact on the variation of the spatial distribution patterns of S. chamaejasme. The spatial distribution pattern varied from a random distribution to a clumped distribution and then returned to a random distribution with increasing coverage (from 31-40% to 71-80%), and this may indicate that the S. chamaejasme patches experienced patch formation and extension and merged with each other.


Subject(s)
Thymelaeaceae/physiology , China , Ecosystem , Grassland , Plants, Toxic/physiology
9.
J Asian Nat Prod Res ; 20(2): 109-116, 2018 Feb.
Article in English | MEDLINE | ID: mdl-28480740

ABSTRACT

A new p-coumaroylated santalane-type sesquiterpenoid, 8-p-coumaroyl-α-santalene (1), a new p-coumaroylated oplopanane-type sesquiterpenoid, 8-ß-p-coumaroyl-oplopanone (2), and three known p-coumaroylated humulene-type sesquiterpenoids (3-5) were isolated from the ethanol extract of the whole herbs of Pilea cavaleriei. Their structures were elucidated based on the combination of 1D and 2D NMR and HRMS methods. Compound 2 was found to show anti-tuberculosis activity with MIC of 16 µg/ml.


Subject(s)
Sesquiterpenes/isolation & purification , Urticaceae/chemistry , Drugs, Chinese Herbal/chemistry , Molecular Structure , Monocyclic Sesquiterpenes , Nuclear Magnetic Resonance, Biomolecular , Polycyclic Sesquiterpenes , Sesquiterpenes/chemistry , Sesquiterpenes/pharmacology
10.
Fitoterapia ; 119: 100-107, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28408268

ABSTRACT

Three new sesquiterpene glycosides (1-3), three new glycerol glycosides (4-6), two new alkaloids (7-8), together with seven known compounds (9-15) all of which were isolated from the genus Pilea for the first time, were isolated from the whole plants of Pilea cavaleriei Levl subsp. cavaleriei. Their structures were determined by extensive spectroscopic techniques and chemical methods. The cytotoxic activity of the isolated compounds was evaluated against four cancer cell lines, and none of the tested compounds caused a significant reduction of the cell number.


Subject(s)
Alkaloids/chemistry , Glycosides/chemistry , Sesquiterpenes/chemistry , Urticaceae/chemistry , Alkaloids/isolation & purification , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Cell Line, Tumor , Glycosides/isolation & purification , Humans , Molecular Structure , Sesquiterpenes/isolation & purification
11.
Phys Chem Chem Phys ; 18(9): 6893-900, 2016 Mar 07.
Article in English | MEDLINE | ID: mdl-26879071

ABSTRACT

Electrochemical cycling stabilities were compared for undoped and Al/Co dual-doped spinel LiMn2O4 synthesized by solid state reactions. We observed the suppression of particle fracture in Al/Co dual-doped LiMn2O4 during charge/discharge cycling and its distinguishable particle morphology with respect to the undoped material. Systematic first-principles calculations were performed on undoped, Al or Co single-doped, and Al/Co dual-doped LiMn2O4 to investigate their structural differences at the atomistic level. We reveal that while Jahn-Teller distortion associated with the Mn(3+)O6 octahedron is the origin of the lattice strain, the networking -i.e. the distribution of mixed valence Mn ions - is much more important to release the lattice strain, and thus to alleviating particle cracking. The calculations showed that the lattice mismatching between Li(+) intercalation and deintercalation of LiMn2O4 can be significantly reduced by dual-doping, and therefore also the volumetric shrinkage during delithiation. This may account for the near disappearance of cracks on the surface of Al/Co-LiMn2O4 after 350 cycles, while some obvious cracks have developed in undoped LiMn2O4 at similar particle size even after 50 cycles. Correspondingly, Al/Co dual-doped LiMn2O4 showed a good cycling stability with a capacity retention of 84.1% after 350 cycles at a rate of 1C, 8% higher than the undoped phase.

12.
ACS Appl Mater Interfaces ; 7(42): 23605-14, 2015 Oct 28.
Article in English | MEDLINE | ID: mdl-26451678

ABSTRACT

An electrolyte additive, p-toluenesulfonyl isocyanate (PTSI), is evaluated in our work to overcome the poor cycling performance of spinel lithium titanate (Li4Ti5O12) lithium-ion batteries. We find that the cycling performance of a Li/Li4Ti5O12 cell with 0.5 wt % PTSI after 400 cycles is obviously improved. Remarkably, we also find that a solid electrolyte interface (SEI) film is formed about 1.2 V, which has higher potential to generate a stable SEI film than do carbonate solvents in the voltage range of 3.0-0 V. The stable SEI film derived from PTSI can effectively suppress the decomposition of electrolyte, HF generation, interfacial reaction, and LiF formation upon cycling. These observations are explained in terms of PTSI including SO3. The S═O groups can delocalize the nitrogen core, which acts as the weak base site to hinder the reactivity of PF5. Hence, HF generation and LiF formation are suppressed.

13.
Yao Xue Xue Bao ; 49(5): 661-5, 2014 May.
Article in Chinese | MEDLINE | ID: mdl-25151738

ABSTRACT

The mechanism of biological actions of quercetin was studied by using metabolomic method and biomolecular network. HPLC-MS was used to analyze the serum metabolome in rats of blank group and quercetin administration group rats, and MS data were processed by MATLAB software. With multivariate statistical analysis of serum metabolite profiles, a clear separation among blank group and quercetin administration group was achieved, potential biomarkers were selected according to the parameters of variable importance in the projection (VIP) and identified according to MS information and database retrieval. Four compounds, related enzymes, action targets and metabolic pathways had been confirmed, namely retinoic acid and RARbeta, arachidonate and COX-2, 3, 5-diodotyrosine and TPO, uridine diphosphate glucose and PDEs. The mechanism of quercetin enhancing ability of retinoic acid on the induction of RARbeta, activating TPO, using as COX-2 and PDEs inhibitor was approved by biomolecular network and related literatures. In this study, a mechanism of multiple biological actions of quercetin was evaluated at the level of the biomolecular network, metabolomics and biomolecular network can be used to investigate the biological effects mechanism of quercetin, which provided a new method to further revealing mechanism of drug action.


Subject(s)
Antioxidants/pharmacology , Metabolomics , Quercetin/pharmacology , Animals , Biomarkers/blood , Chromatography, Liquid , Mass Spectrometry , Metabolic Networks and Pathways , Metabolome , Multivariate Analysis , Rats
14.
J Asian Nat Prod Res ; 16(6): 565-73, 2014.
Article in English | MEDLINE | ID: mdl-24911100

ABSTRACT

Five new phenolic glycosides, 2-hydroxy-(2'E)-prenyl benzoate-2,4'-di-O-ß-D-glucopyranoside (1), 2-hydroxy-(2'E)-prenyl benzoate-2-O-α-L-arabinopyranosyl-(1 → 6)-ß-D-glucopyranoside (2), 4-methylphenol-1-O-α-L-rhamnopyranosyl-(1 → 6)-ß-D-glucopyranoside (3), 4-methylphenol-1-O-α-L-arabinopyranosyl-(1 → 6)-ß-D-glucopyranoside (4), and 3,5-dimethoxyphenol-1-O-ß-D-apiofuranosyl-(1 → 2)-ß-D-glucopyranoside (5), together with six known glycosides (6-11), were isolated from the n-BuOH fraction of the EtOH extract of Pilea cavaleriei Levl subsp. cavaleriei. Their structures were elucidated by extensive spectroscopic analysis, including 1D and 2D NMR spectroscopy as well as HR-ESI-MS, and chemical evidences. All these compounds were isolated from the genus Pilea for the first time.


Subject(s)
Drugs, Chinese Herbal/isolation & purification , Glycosides/isolation & purification , Phenols/isolation & purification , Urticaceae/chemistry , Drugs, Chinese Herbal/chemistry , Glycosides/chemistry , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Phenols/chemistry , Stereoisomerism
15.
Biotechnol Biofuels ; 7(1): 5, 2014 Jan 08.
Article in English | MEDLINE | ID: mdl-24401161

ABSTRACT

BACKGROUND: Clostridium acetobutylicum can propagate on fibrous matrices and form biofilms that have improved butanol tolerance and a high fermentation rate and can be repeatedly used. Previously, a novel macroporous resin, KA-I, was synthesized in our laboratory and was demonstrated to be a good adsorbent with high selectivity and capacity for butanol recovery from a model solution. Based on these results, we aimed to develop a process integrating a biofilm reactor with simultaneous product recovery using the KA-I resin to maximize the production efficiency of biobutanol. RESULTS: KA-I showed great affinity for butanol and butyrate and could selectively enhance acetoin production at the expense of acetone during the fermentation. The biofilm reactor exhibited high productivity with considerably low broth turbidity during repeated batch fermentations. By maintaining the butanol level above 6.5 g/L in the biofilm reactor, butyrate adsorption by the KA-I resin was effectively reduced. Co-adsorption of acetone by the resin improved the fermentation performance. By redox modulation with methyl viologen (MV), the butanol-acetone ratio and the total product yield increased. An equivalent solvent titer of 96.5 to 130.7 g/L was achieved with a productivity of 1.0 to 1.5 g · L-1 · h-1. The solvent concentration and productivity increased by 4 to 6-fold and 3 to 5-fold, respectively, compared to traditional batch fermentation using planktonic culture. CONCLUSIONS: Compared to the conventional process, the integrated process dramatically improved the productivity and reduced the energy consumption as well as water usage in biobutanol production. While genetic engineering focuses on strain improvement to enhance butanol production, process development can fully exploit the productivity of a strain and maximize the production efficiency.

16.
Zhongguo Zhong Yao Za Zhi ; 37(17): 2581-4, 2012 Sep.
Article in Chinese | MEDLINE | ID: mdl-23236755

ABSTRACT

OBJECTIVE: To investigate chemical constituents from folk herb Pilea cavaleriei subsp. cavaleriei. METHOD: The compounds were separated and purified by silica gel, Sephadex LH-20 and the like. The structures were identified by spectral methods such as (1)H, (13)C-NMR and MS. RESULT: Seventeen compounds were isolated and identified as benzoic acid (1), 4-hydroxy benzalde-hyde (2), coumaric acid(3), protocatechuic acid (4), gallic acid (5), 4-hydroxy benzoic acid (6), 3-indole carboxaldehyde (7), 3-indole carbo-xylicacid (8), 4-methyl-(1,2,3) -triazole(9), uracil(10), nicotinamide (11), (2S,E)-N-[2-hydroxy-2-(4-hydroxy phenyl) ethyl] ferulamide (12), (+) -dehydrovomifoliol (13), hentriantane (14), beta-sitosterol (15), palmitic acid (16), daucossterol (17) , respectively. CONCLUSION: All compounds were obtained from the genus for the first time.


Subject(s)
Drugs, Chinese Herbal/chemistry , Urticaceae/chemistry , Dextrans/chemistry , Dextrans/isolation & purification , Drugs, Chinese Herbal/isolation & purification , Molecular Structure , Sitosterols/chemistry , Sitosterols/isolation & purification
17.
J Asian Nat Prod Res ; 14(11): 1032-8, 2012.
Article in English | MEDLINE | ID: mdl-22924790

ABSTRACT

A new triterpenoid, 11α,12α-epoxy-3ß-hydroxy-24-nor-olean-4(23)-en-28,13ß-olide (1), and a new glycoside, benzyl 2-O-ß-d-apiofuranosyl-(1 â†’ 2)-ß-d-glucopyranosyl benzoate (2), together with eight known triterpenoids (3-10), were isolated from Pilea cavaleriei subsp. cavaleriei. Their structures were established on the basis of spectroscopic analysis including HR-ESI-MS, 1D NMR, and 2D NMR techniques. All compounds showed no anti-hepatitis C virus activity.


Subject(s)
Drugs, Chinese Herbal/isolation & purification , Glycosides/isolation & purification , Triterpenes/isolation & purification , Urticaceae/chemistry , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Glycosides/chemistry , Glycosides/pharmacology , Hepacivirus/drug effects , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Triterpenes/chemistry , Triterpenes/pharmacology
18.
J Chem Phys ; 122(22): 224303, 2005 Jun 08.
Article in English | MEDLINE | ID: mdl-15974664

ABSTRACT

The dipole-forbidden transition of D1delta <-- X1sigma+ of CO has been observed by a fast electron-energy-loss spectrometer at a large scattering angle of 7 degrees and with an energy resolution of 60 meV. The energy levels and the relative intensity distribution of D1delta, v'=9-25 have been determined, most of the results are reported for the first time.

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