Urinary nuclear magnetic resonance spectroscopy of a Bangladeshi cohort with hepatitis-B hepatocellular carcinoma: A biomarker corroboration study

AIM: To establish if a distinct urinary metabolic profile could be identified in Bangladeshi hepatitis-B hepatocellular carcinoma (HCC) patients compared to cirrhosis patients and controls.METHODS: Urine samples from 42 Bangladeshi patients with HCC (39 patients with hepatitis-B HCC), 47 with cirrhosis on a background of hepatitis B, 46 with chronic hepatitis B, and seven ethnically-matched healthy controls were analyzed using nuclear magnetic resonance (NMR) spectroscopy. A full dietary and medication history was recorded for each subject. The urinary NMR data were analyzed using principal component analysis (PCA) and orthogonal partial least squared discriminant analysis (OPLS-DA) techniques. Differences in relative signal levels of the most discriminatory metabolites identified by PCA and OPLS-DA were compared between subject groups using an independent samples Kruskal-Wallis one-way analysis of variance (ANOVA) test with all pairwise multiple comparisons. Within the patient subgroups, the Mann-Whitney U test was used to compare metabolite levels depending on hepatitis B e-antigen (HBeAg) status and treatment with anti-viral therapy. A Benjamini-Hochberg adjustment was applied to acquire the level of significance for multiple testing, with a declared level of statistical significance of P < 0.05.RESULTS: There were significant differences in age (P < 0.001), weight (P < 0.001), and body mass index (P < 0.001) across the four clinical subgroups. Serum alanine aminotransferase (ALT) was significantly higher in the HCC group compared to controls (P < 0.001); serum α-fetoprotein was generally markedly elevated in HCC compared to controls; and serum creatinine levels were significantly reduced in the HCC group compared to the cirrhosis group (P = 0.004). A three-factor PCA scores plot showed clustering of the urinary NMR spectra from the four subgroups. Metabolites that contributed to the discrimination between the subgroups included acetate, creatine, creatinine, dimethyamine (DMA), formate, glycine, hippurate, and trimethylamine-N-oxide (TMAO). A comparison of relative metabolite levels confirmed that carnitine was significantly increased in HCC; and creatinine, hippurate, and TMAO were significantly reduced in HCC compared to the other subgroups. HBeAg negative patients showed a significant increase in creatinine (P = 0.001) compared to HBeAg positive patients in the chronic hepatitis B subgroup, whilst HBeAg negative patients showed a significant decrease in DMA (P = 0.004) in the cirrhosis subgroup compared to HBeAg positive patients. There were no differences in metabolite levels in HCC patients who did or did not receive antiviral treatment.CONCLUSION: Urinary NMR changes in Bangladeshi HCC were identified, corroborating previous findings from Egypt and West Africa. These findings could form the basis for the development of a cost-effective HCC dipstick screening test.

Using Nuclear Magnetic Resonance and MS/MS Spectroscopy for the Identification of Brodimoprim Metabolites in Rat Urine

Eight metabolites of brodimoprim (BDP) in rat urine were detected by NMR and ESIMS/MS. They were demethyl-BDP glucuronide, demethyl-BDP sulfurate, demethyl-BDP glucuronide sulfurate, alpha -hydroxyl-BDP, alpha -hydroxyl-BDP glucuronide, BDP sulfurate, N-oxide-BDP sulfurate, and alpha -hydroxyl-N-oxide-BDP sulfurate. All the sulfurates are reported for the first time.

Nuclear magnetic resonance spectroscopy is highly sensitive for lipid-soluble metabolites

Although the water-soluble metabolite profile of human mesenchymal stem cells is known, the lipid profile still needs further investigation. In this study, methanol-chloroform was used to extract lipid-soluble metabolites and perchloric acid was used to extract water-soluble metabolites. Fur- thermore, a dual phase extraction method using methanol-chloroform and water was used to obtain both water and lipid fractions simultaneously. All metabolite extractions were analyzed on a 9.4T high-resolution nuclear magnetic resonance spectrometer. Metabolite resonance peaks were as- signed in the acquired spectra according to the chemical shift, and the extraction efficiency of dif- ferent methods was compared. Results showed that in the spectra of water-soluble extracts, major metabolites comprised low molecular weight metabolites, including lactate, acetic acid, fatty acids, threonine, glutamic acid, creatine, choline and its derivatives, while in the spectra of lipid-soluble extracts, most metabolites were assigned to fatty acids. Among the different extraction procedures, perchloric acid was more efficient in extracting water-soluble metabolites and methanol-chloroform was efficient in extracting organic components compared with the dual phase extraction method. Nuclear magnetic resonance spectroscopy showed that as low as 0.7 mg organic yield was enough to obtain clear resonance peaks, while about 6.0 mg water-soluble yield was needed to obtain rela- tively favorable spectral lines. These results show that the efficiency of extracting water and lipid fractions is higher using perchloric acid and methanol-chloroform compared with dual phase ex- traction and that nuclear magnetic resonance spectroscopy is highly sensitive for analyzing lipid-soluble extracts.

Conformational Analysis of γ-Butyrolactones by Nuclear Magnetic Resonance Spectroscopy

A series of trans-4, 5-disubstituted -γ-butyrolactones are found to assume two different envelope conformations by means of nuclear magnetic resonance spectroscopy.

7.0T nuclear magnetic resonance evaluation of the amyloid beta(1–40) animal model of Alzheimer's disease: comparison of cytology verification

3.0T magnetic resonance spectroscopic imaging brain function in Alzheimer＇s disease. However, is a commonly used method in the research ot the role of 7.0T high-field magnetic resonance spectroscopic imaging in brain function of Alzheimer＇s disease remains unclear. In this study, 7.0T magnetic resonance spectroscopy showed that in the hippocampus of Alzheimer＇s disease rats, the N-acetylaspartate wave crest was reduced, and the creatine and choline wave crest was elevated. This finding was further supported by hematoxylin-eosin staining, which showed a loss of hippocampal neurons and more glial cells. Moreover, electron microscopy showed neuronal shrinkage and mitochondrial rupture, and scanning electron microscopy revealed small size hippocampal synaptic vesicles, incomplete synaptic structure, and reduced number. Overall, the results revealed that 7.0T high-field nuclear magnetic resonance spectroscopy detected the lesions and functional changes in hippocampal neurons of Alzheimer＇s disease rats in vivo, allowing the possibility for assessing the success rate and grading of the amyloid beta （1-40） animal model of Alzheimer＇s disease.

Metabolomic studies of human gastric cancer:Review

Metabolomics is a field of study in systems biology that involves the identification and quantification of metabolites present in a biological system. Analyzing metabolic differences between unperturbed and perturbed networks, such as cancerous and noncancerous samples, can provide insight into underlying disease pathology, disease prognosis and diagnosis. Despite the large number of review articles concerning metabolomics and its application in cancer research, biomarker and drug discovery, these reviews do not focus on a specific type of cancer. Metabolomics may provide biomarkers useful for identification of early stage gastric cancer, potentially addressing an important clinical need. Here, we present a short review on metabolomics as a tool for biomarker discovery in human gastric cancer, with a primary focus on its use as a predictor of anticancer drug chemosensitivity, diagnosis, prognosis, and metastasis.

Serum and urine metabolomic fingerprinting in diagnostics of inflammatory bowel diseases

AIM: To evaluate the utility of serum and urine metabolomic analysis in diagnosing and monitoring of inflammatory bowel diseases (IBD).

Influence of climate warming and nitrogen deposition on soil phosphorus composition and phosphorus availability in a temperate grassland,China

Climate warming and nitrogen （N） deposition change ecosystem processes, structure, and functioning whereas the phosphorus （P） composition and availability directly influence the ecosystem structure under condi- tions of N deposition. In our study, four treatments were designed, including a control, diurnal warming （DW）, N deposition （ND）, and combined warming and N deposition （WN）. The effects of DW, ND, and WN on P composition were studied by 3~p nuclear magnetic resonance （3~p NMR） spectroscopy in a temperate grassland region of China. The results showed that the N deposition decreased the soil pH and total N （TN） concentration but increased the soil OIsen-P concentration. The solution-state 31p NMR analysis showed that the DW, ND and WN treatments slightly decreased the proportion of orthophosphate and increased that of the monoesters. An absence of myo-inositol phosphate in the DW, ND and WN treatments was observed compared with the control. Furthermore, the DW, ND and WN treatments significantly decreased the recovery of soil P in the NaOH-EDTA solution by 17%-20%. The principal component analysis found that the soil pH was positively correlated with the P recovery in the NaOH-EDTA solution. Therefore, the decreased soil P recovery in the DW and ND treatments might be caused by an indirect influence on the soil pH. Additionally, the soil moisture content was the key factor limiting the available P. The positive correlation of total carbon （TC） and TN with the soil P composition indicated the influence of climate warming and N deposition on the biological processes in the soil P cycling.

Metabolomics profile in gastrointestinal cancers:Update and future perspectives

Despite recent progress in diagnosis and therapy,gastrointestinal(GI)cancers remain one of the most important causes of death with a poor prognosis due to late diagnosis.Serum tumor markers and detection of occult blood in the stool are the current tests used in the clinic of GI cancers;however,these tests are not useful as diagnostic screening since they have low specificity and low sensitivity.Considering that one of the hallmarks of cancer is dysregulated metabolism and metabolomics is an optimal approach to illustrate the metabolic mechanisms that belong to living systems,is now clear that this-omics could open a new way to study cancer.In the last years,nuclear magnetic resonance(NMR)metabolomics has demonstrated to be an optimal approach for diseases’diagnosis nevertheless a few studies focus on the NMR capability to find new biomarkers for early diagnosis of GI cancers.For these reasons in this review,we will give an update on the status of NMR metabolomic studies for the diagnosis and development of GI cancers using biological fluids.

Determination of chlorogenic acid in traditional Chinese prescription Shuanghuanglian capsule using quantitative nuclear magnetic resonance spectroscopy in combination with solid phase extraction

The determination method of chlorogenic acid in traditional Chinese prescription Shuanghuanglian capsule was established by using quantitative nuclear magnetic resonance spectroscopy(q NMR) in combination with solid phase extraction(SPE). As the capsule’s main active component, chlorogenic acid comes from the extraction of Chinese herb medicine Flos Lonicerae. The chlorogenic acid in capsule was ultrasonically extracted at room temperature using pure water as solvent. The extracting solution was enriched and cleaned using HC-C18 SPE cartridge. The effect of ultrasonic extraction, sample pretreatment conditions via SPE and q NMR experimental conditions were investigated. The q NMR experiment conditions were selected using deuterated DMSO as solvent, calibrated 1,4-phthalaldehyde as internal standard, and P1(pulse width) = 14.4 μs, d1(pulse delay time) = 1 s, NS(number of scan) = 512. The 1 H NMR peaks of δ 6.138–6.182(H-8’, d, 1 H) of chlorogenic acid was chosen as the quantitative peaks. Method validation was performed, including precision(the intra-day RSD = 1.2% and the inter-day RSD = 1.5%), linearity(correlation coefficient r>0.9999), LOD(0.0017 mg/g) and LOQ(0.079 mg/g). The recovery of the SPE-q NMR was within the range of 100.2%–103.2%. The result showed that the method was stable, accurate and reliabile. Determined by the method, the chlorogenic acid in a real Shuanghuanglian capsule was within the range of 9.68–10.35 mg/g.

Grade classification of neuroepithelial tumors using high-resolution magic-angle spinning proton nuclear magnetic resonance spectroscopy and pattern recognition

Clinical data have shown that survival rates vary considerably among brain tumor patients,according to the type and grade of the tumor.Metabolite profiles of intact tumor tissues measured with high-resolution magic-angle spinning proton nuclear magnetic resonance spectroscopy (HRMAS 1H NMRS) can provide important information on tumor biology and metabolism.These metabolic fingerprints can then be used for tumor classification and grading,with great potential value for tumor diagnosis.We studied the metabolic characteristics of 30 neuroepithelial tumor biopsies,including two astrocytomas (grade I),12 astrocytomas (grade II),eight anaplastic astrocytomas (grade III),three glioblastomas (grade IV) and five medulloblastomas (grade IV) from 30 patients using HRMAS 1H NMRS.The results were correlated with pathological features using multivariate data analysis,including principal component analysis (PCA).There were significant differences in the levels of N-acetyl-aspartate (NAA),creatine,myo-inositol,glycine and lactate between tumors of different grades (P<0.05).There were also significant differences in the ratios of NAA/creatine,lactate/creatine,myo-inositol/creatine,glycine/creatine,scyllo-inositol/creatine and alanine/creatine (P<0.05).A soft independent modeling of class analogy model produced a predictive accuracy of 87% for high-grade (grade III-IV) brain tumors with a sensitivity of 87% and a specificity of 93%.HRMAS 1H NMR spectroscopy in conjunction with pattern recognition thus provides a potentially useful tool for the rapid and accurate classification of human brain tumor grades.

Structural Description of Polyaromatic Nucleus in Residue

The proton nuclear magnetic resonance spectroscopy （1H-NMR）, the synchronous fluorescence spectrometry （SFS） and the ruthenium ions catalyzed oxidation （RICO） method were used to determine the chemical structure of polyaromatic nucleus in Oman residue fractions. The results of 1H-NMR analyses showed that the average numbers of aromatic rings in the aromatics, resins and asphaltenes units were 3.2, 5.6 and 8.2, respectively. SFS was used to investigate the distribution of aromatic rings in residue fractions, the main distribution range of aromatic rings in aromatics, resins and asphaltenes were 3 4 rings, 3--5 rings and more than 5 rings, respectively. The aromatic network in residue fractions was oxidized to produce numerous carboxylic acids. The types and content of benzenepolycarboxylic acids, such as phthalic acid, benzenetricarboxylic acids, benzenetetracarboxylic acids, benzenepentacarboxylic acid and benzenehexacarboxylic acid disclosed the condensed types of aromatic nuclei in the core. The biphenyl fraction （BIPH）, the cata-condensed fraction （CATA）, the peri-condensed fraction （PERI） and the condensed index （BCI） were calculated based on the benzenepolycarboxylic acids formed. The results implied that there was less biphenyl type structures in all residue fractions. The aromatics fraction was almost composed of the cata-condensed type system, and the asphaltenes fraction was wholly composed of the peri-condensed type system, while in the resins fraction co-existed the two types, herein the peri-con- densed type was predominant over the cata-condensed type. Based on the analytical results obtained in the study, the components --aromatics, resins and asphaltenes -- were given the likely structural models.

Soil Phosphorus Composition and Phosphatase Activities along Altitudes of Alpine Tundra in Changbai Mountains,China

Alpine tundra ecosystems have specific vegetation and environmental conditions that may affect soil phosphorus （P） composition and phosphatase activities. However, these effects are poody understood. This study used NaOH-EDTA extraction and solution ^31P nuclear magnetic resonance （NMR） spectroscopy to determine soil P composition and phosphatase activities, including acid phosphomonoesterase （AcP）, phosphodiesterase （PD） and inorganic pyrophosphatase （IPP）, in the alpine tundra of the Changbai Mountains at seven different altitudinal gradients （i.e., 2000 m, 2100 m, 2200 m, 2300 m, 2400 m, 2500 m, and 2600 m）. The results show that total P （TP）, organic P （OP）, OP/TP, NaOH-EDTA extracted P and AcP, PD, and IPP activities over the altitude range of 2500-2600 m are significantly lower than those below 2400 m. The dominant extracted form of P is OP （73%0-83%） with a large proportion of monoesters （65%0-72%）, whereas inorganic P is present in lower proportions （17%-27%）. The activity of AcP is significantly positively correlated with the contents of soil OP, total carbon （TC）, total nitrogen （TN）, and TP （P 〈 0.05）, indicating that the AcP is a more sensitive index for responding P nutrient storage than PD and IPP. Soil properties, P composition, and phosphatase activities decrease with increased altitude and soil pH. Our results indicate that the distribution of soil P composition and phosphatase activities along altitude and AcP may play an important role in P hydrolysis as well as have the potential to be an indicator of soil quality.

NMR Structural Studies on Alamethicin Dimers

N labeled alamethicin dimer was synthesized. The structure and dynamics of alamethicin dimers were studied with nuclear magnetic resonance (NMR) spectroscopy. The data from 15 N-labeled alamethicin dimer suggest little differences in conformation between the dimer and monomer in the Aib 1 -Pro 14 region. Significant difference in the conformation of the C-terminus are manifest in the NH chemical shifts in the Val 15 -Pho 20 region.

Application of Nuclear Magnetic Resonance Spectroscopy in Metabolic Differences and Potential Biomarkers Screening in Rats with Polycystic Ovary Syndrome

Objective:To investigate the application of nuclear magnetic resonance(NMR)spectroscopy in the exploration of metabolic differences and potential biomarkers in rats with polycystic ovary syndrome(PCOS).Methods:PCOS model was established by subcutaneous injection of deoxy-epiandrosterone(DHEA)in 30 rats randomly selected from sixty 23-day-old SPF female SD rats and set as PCOS group,and the other 30 rats were subcutaneously injected with oil and set as oil group,PCOS model was successfully established by 20 days of injection in both groups,and blood samples were taken for biochemical detection and NMR detection analysis.At the same time,the body weight,ovarian weight and HE staining results of rats in both groups were observed.Results:Compared with the oil group,the rats in the PCOS group had higher body weight,ovarian weight and serum LH,FPG,T,E_(2)and HOMA levels,and lower serum FSH and INS levels(P<0.05).In the oil group,a large number of corpus luteum,follicles at different stages of development and multi-layered granulosa cells were observed under microscope after HE staining;In PCOS group,corpus luteum formation was only visible in some ovaries after HE staining,and all of them showed a higher number of cystic dilated follicles,thickened follicular membrane layer and less granulosa cell layer.The PCA score map of serum NMR profiles of the rats in the two groups showed that the samples were separated between the PCOS group and the oil group but the samples were aggregated within the group,and there were differences in blood metabolism between the PCOS model constructed by surface DHEA and the rats in the oil group.A total of 70 metabolites were detected in the serum samples of rats in the two groups by NMR technique,and 31 of them were different between the two groups,which can be classified into glucose,lipid,and and amino acid.There were significant differences in three metabolites,including isoleucine,valine,and creatine(P<0.01).Conclusion:There are differences in carbohydrate,lipid and amino acid metabolism between normal rats and PCOS rats,among which the levels of serum isoleucine,valine and creatine can be used as important biomarkers in the diagnosis of PCOS.

QUALITY CONTROL OF HERBAL MATERIAL:ALTERNATIVE AND/OR COMPLEMENTARY TOOLS BASED ON BIOSENSORS, DNA PROFILING, NEAR INFRARED SPECTROSCOPY AND NUCLEAR MAGNETIC RESONANCE

Phar.Eur.Herbal Drug(HD)monographs state which aspects have to be considered for quality assurance through the relevant chapters'Definition'.'Characters','Identification','Tests',and'Assay'.Identification of botanical material is achieved by macroscopic and microscopic morphology,generally examined by a trained expert.Content or assay is the most difficult area of

Accurate Quantification of Pure Thiacloprid with Mass Balance and Quantitative H-NMR

Thiacloprid is the first-generation neonicotinoid insecticide,which is widely used in modern agriculture.Reference materials are essential for sound measurement and traceability of results in chemical analysis.Pure thiacloprid was comprehensively assessed by two orthonormal methods:mass balance(MB) and quantitative nuclear magnetic resonance spectroscopy(qNMR).From MB,its mass fraction value was 998.4 mg/g with the subtraction of structurally related impurities(99.87%),inorganic impurities(0.006 mg/g),water(0.33 mg/g),and volatile organic solvent(0 mg/g).To guarantee sufficient separation of structurally related purity,peak purity of thiacloprid was evaluated by five points of UV spectra.From the qNMR of hydrogen,its mass fraction value was 995.7 mg/g which was traced to the internal standard(methyl sulfone).Finally,the certified value was assigned to 997 mg/g,together with an expanded uncertainty of 3 mg/g(k=2).This high-purity thiacloprid can be applied to routine monitoring in agriculture and the food fields.

Spectroscopy of Low-lying States in Odd-Z Odd-A Nuclei Beyond Lead

Low-lying states in odd-Z odd-mass nuclei at the proton drip-line beyond lead have recently been studied through fusion-evaporation reactions using a gas-filled recoil separator. Isomeric 1/2+and13/2+states have been observed in odd-mass astatine and francium nuclei. The systematic behaviour of the level energies of these states have been studied and a similarity between the 1/2+state in astatine and francium has been found. Furthermore, the 13/2+state has been observed in the francium nuclei with an oblate behaviour suggesting a coupling of the i13/2proton to the 2p- 2h intruder excitation.

An atomic electronegative distance vector and carbon-13 nuclear magnetic resonance chemical shifts of alcohols and alkanes

A novel atomic electronegative distance vector (AEDV) has been developed to express the chemical environment of various chemically equivalent carbon atoms in alcohols and alkanes. Combining AEDV and γ parameter, four five-parameter linear relationship equations of chemical shift for four types of carbon atom are created by using multiple linear regression. Correlation coefficients are R = 0.9887, 0.9972, 0.9918 and 0.9968 end roots of mean square error are RMS = 0.906, 0. 821, 1.091 and 1.091 of four types of carbons, i.e., type 1, 2, 3, and 4 for primary, secondary, tertiary, and quaternary carbons, respectively. The stability and prediction capacity for external samples of four models have been tested by cross-validation.

RNA structure determination:From 2D to 3D

RNA molecules serve a wide range of functions that are closely linked to their structures.The basic structural units of RNA consist of single-and double-stranded regions.In order to carry out advanced functions such as catalysis and ligand binding,certain types of RNAs can adopt higher-order structures.The analysis of RNA structures has progressed alongside advancements in structural biology techniques,but it comes with its own set of challenges and corresponding solutions.In this review,we will discuss recent advances in RNA structure analysis techniques,including structural probing methods,X-ray crystallography,nuclear magnetic resonance,cryo-electron microscopy,and small-angle X-ray scattering.Often,a combination of multiple techniques is employed for the integrated analysis of RNA structures.We also survey important RNA structures that have been recently determined using various techniques.