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  • 1
    Language: English
    In: 2015, Vol.10(6), p.e0130822
    Description: The laryngeal chemoreflex (LCR) induces apnea, glottis closure, bradycardia and hypertension in young and maturing mammals. We examined the distribution of medullary nuclei that are activated by the LCR and used immunofluorescent detection of Fos protein as a cellular marker for neuronal activation to establish that the medullary catecholaminergic and serotoninergic neurons participate in the modulation of the LCR. The LCR was elicited by the infusion of KCl-HCl solution into the laryngeal lumen of adult rats in the experimental group, whereas the control group received the same surgery but no infusion. In comparison, the number of regions of Fos-like immunoreactivity (FLI) that were activated by the LCR significantly increased in the nucleus of the solitary tract (NTS), the vestibular nuclear complex (VNC), the loose formation of the nucleus ambiguus (AmbL), the rostral ventral respiratory group (RVRG), the ventrolateral reticular complex (VLR), the pre-Bötzinger complex (PrBöt), the Bötzinger complex (Böt), the spinal trigeminal nucleus (SP5), and the raphe obscurus nucleus (ROb) bilaterally from the medulla oblongata. Furthermore, 12.71% of neurons with FLI in the dorsolateral part of the nucleus of the solitary tract (SolDL) showed tyrosine hydroxylase-immunoreactivity (TH-ir, catecholaminergic), and 70.87% of neurons with FLI in the ROb were serotoninergic. Our data demonstrated the distribution of medullary nuclei that were activated by the LCR, and further demonstrated that catecholaminergic neurons of the SolDL and serotoninergic neurons of the ROb were activated by the LCR, indicating the potential central pathway of the LCR.
    Keywords: Research Article
    E-ISSN: 1932-6203
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  • 2
    Language: English
    In: European Journal of Pharmacology, Oct 5, 2015, Vol.764, p.353(10)
    Description: To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ejphar.2015.06.057 Byline: Qinsheng Dai, Qian Yin, Yikai Zhao, Ruichen Guo, Zhiyu Li, Shiping Ma, Na Lu Abstract: Study of the mechanisms of apoptosis in tumor cells is an important field of tumor therapy and cancer molecular biology. We recently established that III-10, a new flavonoid with a pyrrolidinyl and a benzyl group substitution, exerted its anti-tumor effect via inducing differentiation of human U937 leukemia cells. In this study, we demonstrated that III-10 induced cell apoptosis in human hepatocellular carcinoma cells. The activation of caspase-3, caspase-9, and the increased expression ratio of Bax/Bcl-2 were detected in III-10-induced apoptosis. Z-VAD-FMK, a pan-caspase inhibitor, partly attenuated the apoptotic induction of III-10 on both HepG2 and BEL-7402 cells. Furthermore, the increase of intracellular reactive oxygen species levels and the reduction of mitochondria [DELTA][PSI]m were also observed in BEL-7402 and HepG2 cells after the treatment of III-10. Pretreatment with NAC, a reactive oxygen species production inhibitor, partly attenuated the apoptosis induced by III-10 via blocking the reactive oxygen species generation. Our data also showed that III-10 induced the release of cytochrome c and AIF to cytosol followed after the reactive oxygen species accumulation. Moreover, the GSH levels and ATP generation were both inhibited after III-10 treatment. Besides, the MAPK, the downstream effect of reactive oxygen species accumulation including JNK could be activated by III-10, as well as the inactivation of ERK. Collectively, the generation of reactive oxygen species might play an crucial role in III-10-induced mitochondrial apoptosis pathway, provided more stubborn evidence for III-10 as a potent anticancer therapeutic candidate. Author Affiliation: (a) State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, People's Republic of China (b) Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing 210009, People's Republic of China (c) Jiangsu Key Laboratory of Drug Design and Optimization, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, People's Republic of China (d) Xi'an Middle School of Shaanxi Province, Xi'an 710021, People's Republic of China Article History: Received 26 February 2015; Revised 26 June 2015; Accepted 26 June 2015
    Keywords: Flavonoids ; Hepatocellular Carcinoma ; Apoptosis ; Leukemia ; Mitochondrial DNA
    ISSN: 0014-2999
    Source: Cengage Learning, Inc.
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  • 3
    Language: English
    In: European Journal of Pharmacology, 05 October 2015, Vol.764, pp.353-362
    Description: Study of the mechanisms of apoptosis in tumor cells is an important field of tumor therapy and cancer molecular biology. We recently established that III-10, a new flavonoid with a pyrrolidinyl and a benzyl group substitution, exerted its anti-tumor effect via inducing differentiation of human U937 leukemia cells. In this study, we demonstrated that III-10 induced cell apoptosis in human hepatocellular carcinoma cells. The activation of caspase-3, caspase-9, and the increased expression ratio of Bax/Bcl-2 were detected in III-10-induced apoptosis. Z-VAD-FMK, a pan-caspase inhibitor, partly attenuated the apoptotic induction of III-10 on both HepG2 and BEL-7402 cells. Furthermore, the increase of intracellular reactive oxygen species levels and the reduction of mitochondria ΔΨm were also observed in BEL-7402 and HepG2 cells after the treatment of III-10. Pretreatment with NAC, a reactive oxygen species production inhibitor, partly attenuated the apoptosis induced by III-10 via blocking the reactive oxygen species generation. Our data also showed that III-10 induced the release of cytochrome c and AIF to cytosol followed after the reactive oxygen species accumulation. Moreover, the GSH levels and ATP generation were both inhibited after III-10 treatment. Besides, the MAPK, the downstream effect of reactive oxygen species accumulation including JNK could be activated by III-10, as well as the inactivation of ERK. Collectively, the generation of reactive oxygen species might play an crucial role in III-10-induced mitochondrial apoptosis pathway, provided more stubborn evidence for III-10 as a potent anticancer therapeutic candidate.
    Keywords: Apoptosis ; Reactive Oxygen Species ; Mitochondria ; Mmp ; Mapk ; Pharmacy, Therapeutics, & Pharmacology
    ISSN: 0014-2999
    E-ISSN: 1879-0712
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  • 4
    In: Molecular BioSystems, 2012, Vol.8(8), pp.2197-2204
    Description: Epilepsy is a cryptogenic neurological disorder characterized by recurrent seizures which may be precipitated by a variety of endogenous or exogenous factors, and usually occurs many months or years after a precipitating injury. Timely diagnosis and treatment at the early stage of epilepsy are very important for patients to prevent serious lesions and improve the quality of their life. In this study, the metabonomics approach based on the GC-MS technique, multivariate statistical analysis and the metabolism network analysis were applied to investigate metabolic changes in epileptic patients. The outcome of this study suggested that ten endogenous metabolites and five metabolism pathways were mainly involved and showed marked perturbations in epileptic patients. It not only enhances the understanding of the pathology of epilepsy, but also provides an experimental foundation for the therapeutic strategy of epilepsy. Furthermore, this work demonstrates the powerful predictive potential of the metabolic network analysis to neurological disease.
    Keywords: Epilepsy -- Blood ; Gas Chromatography-Mass Spectrometry -- Methods ; Metabolomics -- Methods;
    ISSN: 1742-206X
    E-ISSN: 1742-2051
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  • 5
    Language: English
    In: Current Pharmaceutical Biotechnology, 2017, Vol.18(13), p.1098-1105
    Description: Background: Phenytoin has very challenging pharmacokinetic properties. To prevent its toxicity and ensure efficacy, continuous therapeutic monitoring is required. It is hard to get a simple, accurate, rapid, easily available, economical and highly sensitive assay in one method for therapeutic monitoring of phenytoin. 〈/P〉〈P〉 Objective: The present study is directed towards establishing and validating a simpler, rapid, an accurate, highly sensitive, novel and environment friendly liquid chromatography/mass spectrometry (LC/MS) method for offering rapid and reliable TDM results of phenytoin in epileptic patients to physicians and clinicians for making immediate and rational decision. 〈/P〉〈P〉 Methods: 27 epileptics patients with uncontrolled seizures or suspected of non-compliance or toxicity of phenytoin were selected and advised for TDM of phenytoin by neurologists of Qilu Hospital Jinan, China. The LC/MS assay was used for performing of therapeutic monitoring of phenytoin. The Agilent 1100 LC/MS system was used for TDM. The mixture of Ammonium acetate 5mM: Methanol at (35: 65 v/v) was used for the composition of mobile phase. The Diamonsil C18 (150mm×4.6mm, 5μm) column was used for the extraction of analytes in plasma. The samples were prepared with one step simple protein precipitation method. The technique was validated with the guidelines of International Conference on Harmonisation (ICH). 〈/P〉〈P〉 Results: The calibration curve demonstrated decent linearity within (0.2-20 µg/mL) concentration range with linearity equation, y= 0.0667855 x +0.00241785 and correlation coefficient (R2) of 0.99928. The specificity, recovery, linearity, accuracy, precision and stability results were within the accepted limits. The concentration of 0.2 µg/mL was observed as lower limit of quantitation (LLOQ), which is 12.5 times lower than the currently available enzyme-multiplied immunoassay technique (EMIT) for measurement of phenytoin in epilepsy patients. 〈/P〉〈P〉 Conclusion: A rapid, simple, economical, precise, highly sensitive and novel LC/MS assay has been established, validated and applied successfully in TDM of 27 epileptics patients. It was alarmingly found that TDM results of all these patients were out of safe range except two patients. However, it needs further evaluation. Besides TDM, the stated method can also be applied in bioequivalence, pharmacokinetics, toxicokinetics and pharmacovigilance studies.
    Keywords: Antiepileptic Drugs Epilepsy Lc/Ms Novel Phenytoin Rapid.
    ISSN: 1389-2010
    E-ISSN: 1873-4316
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  • 6
    In: Biomedical Chromatography, March 2014, Vol.28(3), pp.348-353
    Description: Glucuronidation plays critical role in the elimination of bergenin; however the metabolic mechanism of UDP‐glucuronosyltransferases (UGTs) in the process remains to be investigated. In this study, the kinetics of bergenin glucuronidation by pooled human liver microsomes (HLMs) and 12 recombinat UGT isozymes were investigated. The glucuronidation of bergenin can be shown in HLMs with a value of 231.62 ± 14.08 µ and a value of 2.17 ± 0.21 nmol/min/(mg protein). Among the 12 human UGTs investigated, UGT1A1 was identified as the major isoform catalyzing the glucuronidation of bergenin [ value of 200.37 ± 26.73 µ and value of 1.88 ± 0.26 nmol/min/(mg protein)]. The bergenin glucuronosyltransferase activities in HLMs and UGT1A1 were inhibited by phenylbutazone, estradiol and bilirubin. The results demonstrate that bergenin glucuronidation in HLMs is specifically catalyzed by UGT1A1. Copyright © 2013 John Wiley & Sons, Ltd.
    Keywords: Bergenin ; Udp‐Glucuronosyltransferases ; Human Liver Microsomes ; Ugt1a1 ; In Vitro
    ISSN: 0269-3879
    E-ISSN: 1099-0801
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  • 7
    Language: English
    In: Respiratory Physiology & Neurobiology, June 2016, Vol.226, pp.121-127
    Description: The laryngeal adductor reflex (LAR) is a laryngeal protective reflex. Vagal afferent polymodal sensory fibres that have cell bodies in the nodose ganglion, originate in the sub-glottal area of the larynx and upper trachea. These polymodal sensory fibres respond to mechanical or chemical stimuli. The central axons of these sensory vagal neurons terminate in the dorsolateral subnuclei of the tractus solitarius in the medulla oblongata. The LAR is a critical, reflex in the pathways that play a protective role in the process of ventilation, and the sychronisation of ventilation with other activities that are undertaken by the oropharyngeal systems including: eating, speaking and singing. Failure of the LAR to operate properly at any time after birth can lead to SIDS, pneumonia or death. Despite the critical nature of this reflex, very little is known about the central pathways and neurotransmitters involved in the management of the LAR and any disorders associated with its failure to act properly. Here, we review current knowledge concerning the medullary nuclei and neurochemicals involved in the LAR and propose a potential neural pathway that may facilitate future SIDS research.
    Keywords: Laryngeal Adductor Reflex ; Medulla Oblongata ; Pathway ; Neurotransmitter ; Sudden Infant Death Syndrome ; Anatomy & Physiology
    ISSN: 1569-9048
    E-ISSN: 1878-1519
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  • 8
    Language: English
    In: International journal of clinical pharmacology and therapeutics, August 2018, Vol.56(8), pp.387-392
    Description: Tetramethylpyrazine, isolated from Ligusticum wallichii Franch., is widely used for the treatment of cerebrovascular and cardiovascular diseases in China. To assess and compare the pharmacokinetic characteristics and bioequivalence of two tetramethylpyrazine phosphate (TMPP) tablets in healthy Chinese male subjects. 20 healthy male subjects were randomly divided into two groups according to a two-period crossover design test. A single oral dose of 200 mg test or reference tablets was given with a 7-day washout period under fasting conditions. Blood samples were taken at scheduled time points, and the concentration of TMPP was measured by LC-MS. Drug And Statistical Software-Version 2.0 was used to calculate the pharmacokinetic parameters and assess bioequivalence of the two formulations. 20 subjects were enrolled in the study, and none dropped out. The main pharmacokinetic parameters of test and reference formulations were as follows: T1/2 was (1.79 ± 0.82) hours and (1.64 ± 0.52) hours, tmax was (0.76 ± 0.37) hours and (0.94 ± 0.44) hours, Cmax was (961.14 ± 309.64) ng/mL and (1,059.09 ± 350.69) ng/mL, AUC0-12h was (1,744.69 ± 643.49) ng×h/mL and (1,726.32 ± 494.11) ng×h/mL, AUC0-∞ was (1,756.95 ± 643.63) ng×h/mL and (1,740.16 ± 504.89) ng×h/mL, respectively. The relative bioavailability of TMPP tablets was 102.4 ± 26.0%, and no serious adverse events were reported. This single-dose study in healthy Chinese male fasted subjects showed that the TMPP test and reference tablets were bioequivalent.
.
    Keywords: Plant Extracts -- Pharmacokinetics ; Pyrazines -- Pharmacokinetics
    ISSN: 0946-1965
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  • 9
    In: Journal of Analytical Sciences, Methods and Instrumentation, 2012, Vol.02(01), pp.24-28
    Description: A new and simple gas chromatographic (GC) method using flame ionization detector (FID) and mass spectrometry (MS) for the simultaneous determination of eight fatty alcohols in 10 mg policosanol film-coated tablets was established and applied to the quality control (QC) of policosanol film-coated tablets. A DB-35MS capillary column (30 m × 0.32 mm, 0.25 um) was employed for the separation. GC-FID was used to quantitatively analyze the eight ingredients with 1-eicosanol as internal standard, three of which were identified using GC-MS due to the lack of standard. The linearity, accuracy, precision, stability, robustness and sensitivity within the detection limits were evaluated. The average recovery of the method was 96.3% - 100.4% and linearity was (R > 0.999). The average drug content was found to be 96.8% of the labeled amount (10 mg).
    Keywords: Policosanol ; Fatty Alcohols ; Gas Chromatography-Mass Spectrometry ; Gas Chromatography-Hydrogen Flame Ionization Detector;
    ISSN: 2164-2745
    E-ISSN: 2164-2753
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  • 10
    Language: English
    In: Pakistan Journal of Pharmaceutical Sciences, 2018, Vol.31(4 SI), p.1697(4)
    Description: The narrow therapeutic index, non-linear pharmacokinetics and unpredictable absorption require regular therapeutic monitoring of phenytoin. The influence of genetic differences, sex, age and race on the phenytoin plasma levels and its metabolites is well recognized. This study is aimed at evaluating phenytoin plasma drug concentration and its relationship with clinical response, persistent seizures and toxicity in different gender and various age groups of Chinese epileptic patients. This knowledge will help the clinicians in adjusting the drug dosages of phenytoin in various sub-groups of epileptic patients for enhancing the safety, efficacy and minimizing the toxicity of phenytoin. A total of 48 plasma samples of epileptic patients for measuring the plasma phenytoin concentration were received. Only patients displaying persistent seizures or suspected of adverse effects were requested for drug monitoring. All these samples were analyzed for therapeutic drug monitoring with Enzyme-multiplied immunoassay technique. Surprisingly, it was found that majorities (85.5%) of samples were out of the reference range, of which 69% of samples were in sub-therapeutic levels and 16.5% of samples were above therapeutic levels. Only 14.5% of all samples had phenytoin levels in the therapeutic range. The difference in plasma concentration of phenytoin was notably altered in gender and various age groups. Careful attention must be applied to specific gender and particular age group on an individual basis in the interpretation of plasma concentration results, in order to facilitate the modification of doses and develop the best approach in treatment and to obtain the desired clinical response because multiple factors can affect the phenytoin plasma concentration. Through these results, it can be concluded that a good correlation exists between phenytoin plasma concentration and clinical response. Therefore, regular therapeutic monitoring of phenytoin and screening of HLA-A, B, C and DRB1 genotypes before prescribing phenytoin in epileptic patients is essentially required to achieve maximum clinical response and prevent the serious toxicity.
    Keywords: Toxicity ; Phenytoin ; Seizures (Medicine)
    ISSN: 1011-601X
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