The application of HPLC-MS/MS method in the determination of whole blood concentration of Cyclosporine A and AM1 in bone marrow transplant patient / 重庆医学

Objective To develop a HPLC-MS/MS methodfor determination of cyclosporin A(CSA)and AM1 in bone marrow transplant patient,and explore the relationship of CSA and its main metabolite AM1 within individual and between individuals,and provide reliable basis for clinical rational use of monitoring in CSA blood drug concentration.Methods CSD was used as internal standard,and whole blood samples were treated with internal standard methanol precipitated protein.The column was Ultimate XB-C18 with a column temperature of 65 ℃ and the mobile phase was eluted with 0.1％ of formic acid and 2 mmol/L ammonium acetate in water and methanol containing 0.1％ formic acid.The mass spectrometry was detected by electrospray ion trap positive ion mode,MRM scanning,monitoring CSA 1219.9 to 1203.1 m/z,AM1 1236.1 to 1219.1 m/z,CSD 1234.0 to 1217.0 m/z.Results The concentration of CSA was linear in the range of 16 to 1600 ng/mL,Y=0.0143X+0.0213(r=0.9976).The concentration of AM1 was linear in the range of 10～1000 ng/mL,Y=0.00363X-0.00528(r=0.9973).The ratio of CSA to AM1 (AM1/CSA) between individual ranged from 32％ to 356％.The ratio of CSA to AM1 within indiviolual(AM1/CSA) ranged from 27 ％ to 147 ％.Conclusion HPLC-MS/MS method for the simultaneous detection of CSA and AM1 was established.The variation of CSA exists in the bone marrow transplant patient between individuals and within individual;the HPLC-MS/MS method can be used for monitoring of whole blood concentration of CSA in bone marrow transplant patients.

Standardization of Andrographis paniculata, Mitracarpus scaber and Nauclea latifolia Herbal Preparations as per European and Nigerian Drug Regulations.

Background: Herbal drug standardization (HDS) is multidisciplinary with botany and chemistry working together to facilitate decisions on production of herbal medicines. The common reasons for HDS are: i) it creates the need for establishing botanical identity; ii) it is necessary for establishing dosage and iii) it facilitates industrial production and good manufacturing practice (GMP). Aims: To outline the strategies being used to standardize Conavir, Niprd-AM1 and Niprifan and to show that HDS is the ideal strategy for herbal drug development (HDD) from traditional medicines (TMs). Methodology: Relevant data on: i) the regulatory requirements of Europe’s Medicines Evaluation Agency (EMEA) and Nigeria’s National Agency for Food and Drug Administration and Control (NAFDAC) and ii) on Andrographis paniculata (AP), Mitracarpus scaber (MS) and Nauclea latifolia (NL) were reviewed. Crude herbal drugs (CHDs) from aerial parts each of AP and MS and from roots of NL and the active crude extracts (ACEs) derived from them were studied using standard botanical, phytochemical and physicochemical techniques with the aim of standardizing them for production. The ACEs from AP (Conavir) and from NL (Niprd-AM1) were dry water extracts. The ACE from MS (Niprifan) was a dry ethylacetate extract. Results: The regulatory provisions of NAFDAC for herbal preparations were broadly similar to those of EMEA but the latter proved more explicit in many respects. Furthermore, the results on the CHDs and ACEs adequately meet the requirements of the two agencies. Conclusions: The results here provided and those reported elsewhere collectively furnish the data needed for drawing-up the registration dossiers of AP/Conavir, NL/Niprd- AM1 and MS/Niprifan as per EMEA and NAFDAC requirements. But for purposes of further work, it is needful for the GC-MS studies to be amplified and combined with others, so as to facilitate identification of suitable markers and pave the way for studies requiring bioassays.

Phytochemical, Physicochemical and Chromatographic Profiling in Quality Control Systems for Select Herbal Medicines (Conavir and Niprd-AM1).

Background: Conavir, an immunostimulant from aerial parts of Andrographis paniculata (AP) and Niprd-AM1, an antimalarial from roots of Nauclea latifolia (NL), are dry water extracts for capsulation. AP and NL have been in use in Asia and Africa for centuries. Purpose: The study aimed to ascertain the criteria for quality assured production of Conavir and Niprd-AM1. Experimental Details: Procedures of World Health Organization (WHO) were applied to evaluate quality parameters of AP/ Conavir and NL/ Niprd-AM1. Results and Discussion: Conavir is granular, greenish brown, intensely bitter and practically odourless. Tests on AP and Conavir revealed alkaloids, saponins, tannins and terpenoids, but cardiac and cyanogenic glycosides (considered toxic) were not detected. Normal phase TLC of AP and Conavir yielded 5 principal spots each, while the reverse phase TLC yielded 6. HPLC fingerprints of AP, Conavir and a reference standard were reproducible but differed from each other. The GC-MS data of Conavir were consistent with the phytochemical profile of AP. Effect of storage suggested that both AP and Conavir were stable for up to 21 months or more. Niprd-AM1 is granular, yellowish brown and faintly aromatic, with an exciting bitter taste. Both NL and Niprd-AM1 contained alkaloids, saponins, flavonoids and terpenoids, but cardiac and cyanogenic glycosides were not detected. Normal phase TLC of NL yielded 9 principal spots, while Niprd-AM1 yielded 5, but the reverse phase TLC yielded 9 for each. HPLC fingerprints of NL, Niprd- AM1 and a reference standard were reproducible but differed from each other. The GCMS data of Niprd-AM1 were consistent with the phytochemical profile of NL. Most of the quality variables of NL and Niprd-AM1 remained unchanged up to the 39th month of storage. Conclusion: The results are consistent with NIPRD’s intention to file for the registration of Conavir and Niprd-AM1 for use in Nigeria.

Morphological evidences in circumvallate papilla and von Ebners' gland development in mice / 대한해부학회지

In rodents, the circumvallate papilla (CVP), with its underlying minor salivary gland, the von Ebners' gland (VEG), is located on the dorsal surface of the posterior tongue. Detailed morphological processes to form the proper structure of CVP and VEG have not been properly elucidated. In particular, the specific localization patterns of taste buds in CVP and the branching formation of VEG have not yet been elucidated. To understand the developmental mechanisms underlying CVP and VEG formation, detailed histological observations of CVP and VEG were examined using a three-dimensional computer-aided reconstruction method with serial histological sections and pan-Cytokeratins immunostainings. In addition, to define the developmental processes in CVP and VEG formation, we examined nerve innervations and cell proliferation using microinjections of AM1-43 and immunostainings with various markers, including phosphoinositide 3-kinase, Ki-67, PGP9.5, and Ulex europaeus agglutinin 1 (UEA1). Results revealed specific morphogenesis of CVP and VEG with nerve innervations patterns, evaluated by the coincided localization patterns of AM1-43 and UEA1. Based on these morphological and immunohistochemical results, we suggest that nerve innervations and cell proliferations play important roles in the positioning of taste buds in CVP and branching morphogenesis of VEG in tongue development.

Estudio teóricoppara evaluar la conductividad dl polifurano y el efecto de loss sustituyentes sobre la cadena polimérica / Theoretcal studyy to evaluate polyfuranelectrical conductivity and thee substituent effects on the polymeri cchain / Estud teorico paara avaliar a conductvidade do polifurano o e efeitoo dos substituintes sobre a cadeia polmmerica

Una serie de oligómeros de furano y furano sustituido fueron estudiados desde el punto de vista teórico con el objeto de conocer las propiedades electroconductoras de estos compuestos, y su respectiva extrapolación a polímeros, aprovechando la capacidad de la química computacional para proponer y diseñar nuevos materiales y sus posibles propiedades. Se relacionaron las propiedades electrónicas de estos oligómeros tales como la afinidad electrónica (AE), el potencial de ionización (PI), el band-gap (HOMOLUMO), y la relación de éstos con la conductividad; además, se demostró cómo cambia la longitud de los enlaces de los oligómeros al estar cargados; la longitud de los oligómeros de estudio fuer de dos, cuatro, seis y ocho anillos. En este estudio se realizaron cálculos a niveles AM1 y DFT/B3LYP/6-31G (d).

DFT/B3LYP/6-31G (d) calculations were carried out on a series of molecules of furan and substituted furan to observe the type of variables that affect the conductivity of these molecules. In order to propose and design new molecules and its possible properties. The Ionization potential (IP), band-gap (HOMO-LUMO), electronic affinity (EA), was related with its conductivity. It was also shown how change the length of the olygomers bond when the number of the rings is changed from two to four, six and eight.

Uma serie de oligômeros de furano y furano substituido foram estudados teoricamente com a intenção de conhecer as propriedades electro-conductoras desses compostos e sua respectiva extrapolação a polímeros, aproveitando a capacidade da química computacional para propor e desenhar novos materiais e suas possíveis propriedades. Relacionaram-se as propriedades eletrônicas destes oligômeros, tais como a afinidade eletrônica (AE), o potencial de ionização (PI), o band-gap (HOMO-LUMO) e a relação destes com a condutividade, também se demostrou a mudança do comprimento das ligações dos oligômeros ao estar carregados, o comprimento dos oligômeros em estudo foram de dois, quatro, seis e oito anéis. Em este estudo realizaram-se cálculos a níveis AM1 e DFT/B3LYP/6-31G (d).

An Anti-apoptotic Role of NF-κB in TNFα-induced Apoptosis in an Ameloblastoma Cell Line / Oral Science International

Nuclear factor-κB (NF-κB) is involved in the promotion of cell survival in a variety of cell types. The present study focused on the role of NF-κB in TNFα-induced apoptosis in an ameloblastoma. Immunohistochemical staining revealed p65 NF-κB protein to be expressed in ameloblastoma tissues. Furthermore, immunoblotting and immunocytochemistry analyses showed that the stimulation of TNFα in an ameloblastoma cell line (AM-1) induced p65 NF-κB translocation from the cytoplasm to the nucleus, indicating NF-κB activation. These findings were confirmed by an NF-κB luciferase reporter assay, which detected enhanced NF-κB transcription activity of AM-1 cells by TNFα stimulation. Moreover, pretreatment with SN50, a nuclear translocation inhibitor, prior to TNFα stimulation, effectively inhibited TNFα-induced NF-κB activation in AM-1 cells. In order to reveal the role of NF-κB activation during TNFα-induced apoptosis in AM-1 cells, an apoptosis assay was performed, and showed that the potential of TNFα in inducing apoptosis in AM-1 cells was significantly elevated by inhibiting the NF-κB activation. These results suggest that NF-κB plays an anti-apoptotic role in TNFα-induced apoptosis in AM-1 cells.