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生物分析および生物医学ジャーナル

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音量 3, 問題 1 (2011)

研究論文

Formulation and Characterization of Hydralazine Hydrochloride Biodegraded Microspheres for Intramuscular Administration

Basavraj K. Nanjwade, Hiren M. Bechra, Veerendra K. Nanjwade, Ganesh K. Derkar and F. V. Manvi

Objective: To prepare and characterize Albumin microspheres of hydralazine hydrochloride for the treatment of hypertension.

Methods: Albumin microspheres of antihypertensive drug hydralazine hydrochloride were prepared by emulsion cross-linking method by using glutaraldehyde as cross-linking agent. Drug and polymer compatibility was determined by Fourier-Transform Infrared spectroscopy. To determine the effect of polymer concentration and amount of glutaraldehyde, formulations were characterized for their entrapment efficiency, particle size, surface morphology and release behavior. In vivo study was carried out on hypertensive wistar rats.

Key findings: Maximum percentage entrapment efficiency (%EE) was found to be 68.20±1.03 %. Laser particle size analyzer confirmed mean particle size in the range of 31.7 to 39.6μm. In vitro drug release studies showed a biphasic release pattern for all formulations with an initial burst effect followed by slow release for almost 24 hrs.

Conclusion: In vivo study to determine antihypertensive effect of selected formulation strongly correlates with in vitro drug release behavior. The release behavior was significantly regulated by polymer concentration and volume of glutaraldehyde. The study revealed that hydralazine hydrochloride loaded albumin microspheres exhibited prolonged reduction of systolic and diastolic arterial pressure compared to hydralazine hydrochloride solution.

研究論文

The Identification of Functional Non-Synonymous SNP in Human ATPBinding Cassette (ABC), Subfamily Member 7 Gene: Application of Bioinformatics Tools in Biomedicine

Pornlada Nuchnoi, Duangdao Nantakomol, Vasunun Chumchua, Chotiros Plabplueng, Chartchalerm Isarankura-Na-Ayudhya and Virapong Prachayasittikul

The prediction of functional single nucleotide polymorphism (SNP) is promising in modern genetics analysis. Computational biology technology has facilitated an increase in the successful rate of genetic association study and reduced the cost of genotyping. In the present study, we applied various bioinformatics tools for the selection of high potentially functional nsSNP and determined the linkage disequilibrium (LD) structure of ATP-binding cassette transporter member 7 (ABCA7) genes in HapMap populations. Two functional polymorphisms (rs3752233 and rs3752246) were identified on the basis of less protein stability, a low likelihood of mutability, a changing of protein structure and function. Interestingly, a completed LD between rs3752233 (R463H) and rs4147918 (Q1686R) was detected in Utah residents with ancestry from northern and western Europe (CEU) populations. In addition, the difference of the LD pattern between the populations observed highlighted the essential role of the construction of an LD map for designing and interpreting genetic association study. Studies herein convey the empirical guidelines for conduction of ABCA7 genetic association study via bioinformatics and computational application.

研究論文

Nicotinamide Phosphoribosyltransferase in Human Diseases

Li Qin Zhang, Daniel P. Heruth and Shui Qing Ye

Nicotinamide phosphoribosyltransferase (NAMPT) was first reported as a pre-B-cell colony enhancing factor in 1994 with little notice, but it has received increasing attention in recent years due to accumulating evidence indicating that NAMPT is a pleiotropic protein such as a growth factor, a cytokine, an enzyme and a visfatin. Now, NAMPT has been accepted as an official name of this protein. Because of NAMPT’s multiple functions in a variety of physiological processes, their dysregulations have been implicated in the pathogenesis of a number of human diseases or conditions such as acute lung injury, aging, atherosclerosis, cancer, diabetes, rheumatoid arthritis and sepsis. This review will cover the current understanding of NAMPT’s structure and functions with an emphasis on recent progress of nicotinamide phosphoribosyltransferase’s pathological roles in various human diseases and conditions. Future directions on exploring its Terra incognita will be offered in the end.

研究論文

High-performance Liquid Chromatographic Method with Diode Array Detection for Quantification of Haloperidol Levels in Schizophrenic Patients During Routine Clinical Practice

Tarun Jain, Anil Bhandari, Veerma Ram, Sanjay Sharma, Ratendra K Chaudhary and Manish Parakh

Rationale: Till date, very few HPLC methods are available with short run time for monitoring of haloperidol, facilitating large number of sample analysis within short time frame.

Objective: A selective and sensitive reverse phase high-performance liquid chromatographic assay has been developed for monitoring of Haloperidol levels.

Methodology: Chromatogram separation of Haloperidol and Loratidine (Internal Standard) was achieved using C18 column as stationary phase. Mobile phase consists of Acetonitrile and Water (50:50), pH-2.5 with 0.1% Acetic Acid and 0.05 M KHPO4 at a flow rate of 1.6 mL min-1. Detection was carried out at 240 nm using UV-PDA detector. Retention time for Haloperidol and Internal Standard was found to be 2:13 and 3:16 minutes respectively. The method has been validated for linearity, specificity, robustness, stability, accuracy and precision.

Results: Linearity for Haloperidol was in the range of 03-200 ng mL-1. The total run time of analysis was 5 minutes and the lower limits of detection and quantification were 1.0 and 3.0 ng mL-1 respectively. In present study, most of the enrolled patients were clinically stable on Haloperidol (as evident from various rating scales applied during the study period) at therapeutic range of 5-19 ng mL-1 [13.04 ± 4.03]

Discussion: This validated high-performance liquid chromatographic method using a simple mobile phase has been successfully applied for clinical monitoring of Haloperidol in psychiatric patients. The method is economic and its time sparing ability using simple RP-HPLC speaks its utility in clinical and toxicological management over other analytical methods.

研究論文

Mechanisms Mediating the Synergistic Anticancer Effects of Combined γ-Tocotrienol and Celecoxib Treatment

Amit B. Shirode and Paul W. Sylvester

Aim: To characterize the intracellular signaling mechanisms mediating the synergistic anticancer effects of combined ?-tocotrienol and celecoxib treatment in neoplastic +SA mouse mammary epithelial cells in vitro. Methods: +SA mammary tumor cells in different treatment groups were maintained in serum-free defined media containing 10ng/ml EGF as a mitogen and exposed to various doses of ? -tocotrienol and celecoxib alone or in combination. After a 96 hr culture period, cells were collected and whole cell lysates were subjected to Western blot analysis to determine treatment effects on intracellular signaling proteins associated with EGF-dependent mitogenesis and survival, and prostaglandin synthesis and responsiveness. Results: Treatment with high doses of ?-tocotrienol or celecoxib alone inhibited Akt activation and downstream signaling and NF?B activation. Similar treatment with ?-tocotrienol also decreased concentration and activation of ErbB2-4 receptors, whereas celecoxib only inhibited ErbB2-4 receptor activation. In contrast, combined treatment with subeffective doses of ?-tocotrienol and celecoxib resulted in a large decrease ErbB2-4 receptor levels and activation, a decrease in PGE2 levels, and a corresponding increase in prostaglandin EP2 and EP4 receptor levels. Combined treatment also induced an increase in the prostaglandin catabolizing enzyme, PGDH. Conclusion: The synergistic anticancer effects of combined low dose ?-tocotrienol and celecoxib treatment in +SA mammary tumor cells are mediated by COX-2-dependent mechanisms associated with a suppression in PGE2 levels, as well as, COX-2-independent mechanisms associated with a reduction in ErbB2-4 receptor levels, activation, and subsequent reduction in downstream Akt and NF?B mitogenic signaling.

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