Estimation of liquid diffusivities of biosolutes by using diaphragm cell method with defined pore characteristics

Summary Confidence of the diaphragm cell method with a diaphragm having defined pore characteristics was investigated for liquid diffusivity determination of some biosolutes. Diffusivities of NaCl, KCl, glucose, L-Tryptophan, lysozyme, and BSA were estimated. Satisfactory agreements were observed between estimated diffusivities and those in the literature.     

鸣禽控制发声核团和脑干听觉核团及神经通路的综合研究

采用辣根过氧化物酶顺、逆行标记方法对鸣禽鸟蜡嘴雀控制发声的神经核团、脑干听觉核团及神经通路,从外周至中枢逐级进行了追踪研究。结果表明：１．控制发声的神经核团及通路,前脑古纹状体腹内侧粗核是大脑控制发声的重要核团之一,它发出枕中脑后束经端脑前联合呈双侧支配延脑中间核,中间核又发出舌下神经经气管鸣管分支支配鸣肌,中间核同时也接受中脑背内侧核的支配,２．脑干听觉中枢及通路,中脑背外侧核是脑干较高级听觉中枢、初级中枢耳蜗核由角核和前庭外侧核组成,ＮＡ发出以对侧为主的纤维经外侧丘系可直接传入中脑背外侧核形成脑干听觉直接通路。     

The impact of diabetic neuropathy on the distal versus proximal comparison of weakness in lower and upper limb muscles of patients with type 2 diabetes mellitus: a cross-sectional study

Objectives:This study aimed to determine the impact of diabetic neuropathy (dNP) on the distal versus proximal comparison of weakness in lower and upper limb muscles of patients with type 2 Diabetes Mellitus (T2DM).Methods:19 healthy male controls without neuropathy (HC) and 35 male T2DM patients, without dNP (n=8), with sensory dNP (n=13) or with sensorimotor dNP (dNPsm; n=14), were enrolled in this study. Maximal isometric (IM) and isokinetic (IK) muscle strength and IK muscle endurance of the dominant knee, ankle and elbow, and maximal IM handgrip strength were measured by means of dynamometry.Results:Ankle muscle endurance was lower compared to the knee, independently of dNP (p<0.001). Maximal IK ankle muscle strength was also lower compared to the knee, albeit only in dNPsm (p=0.003). No differences were found between maximal IM handgrip and elbow strength.Conclusions:Our results suggest an impact of T2DM -with or without dNP- on lower limb muscle strength more distally than proximally, while this was not observed in the upper limb. The gradient of dNP seemed to be a determining factor for the maximal muscle strength, and not for muscle endurance, in the lower limb.     

Anthracycline antibiotics. Interaction with DNA and nucleosomes and inhibition of DNA synthesis

We report studies of the interaction of four anthracycline antibiotics, iremycin (IM), daunomycin (DM), aclacinomycin A (AM), and violamycin B1 (VM), with naked DNA, nucleosomal core particles, and 175 base pair (bp) nucleosomes lacking histone H1. In all cases the binding strength increases in the order IM less than DM approximately AM less than VM. The binding substrates increased in affinity for the drugs in the following order: core particles less than 175-bp nucleosomes less than DNA. The apparent DNA length increment per drug bound decreases in the progression IM greater than DM greater than AM greater than VM, the same serial order as is characterized by increasing binding affinity. Dichroism amplitude measurements show that for all drugs the long-wavelength absorbance transition moment is tilted by 26-29 degrees relative to the plane perpendicular to the helix axis; this angle probably corresponds to the long axis tilt of the intercalated chromophore. Finally, it was found that the ability of the drugs to inhibit DNA synthesis by Escherichia coli DNA polymerase I increases in the same order as their binding affinity.     

Diffusion of acetophenone and phenethyl alcohol in the calcium-alginate-bakers' yeast-hexane system

The intrabead diffusion coefficients of acetophenone and phenethyl alcohol were measured at 30 degrees C in the triphasic immobilized yeast-water-hexane system. Saccharomyces cerevisiae cells were deactivated with hydrochloric acid and entrapped in calcium-alginate beads. Measurements of dry cell loss during deactivation, shrinkage of the beads during deactivation and the final porosity of the beads were made for various cell loadings. Final concentrations of wet cells in the beads ranged from approximately 0.25 to 0.30 g/mL. Mass transfer in the hexane phase, external to the beads, was eliminated experimentally. The estimated error of 5% to 10% in the diffusion coefficients is within the experimental error associated with the bead method. The effect of significant sampling volumes on the diffusivities was estimated theoretically and accounted for experimentally. The intrabead concentration of acetophenone and phenethyl alcohol was 150 to 800 ppm. The deactivated cells were shown to be impervious to acetophenone so that the measured diffusivities are extracellular parameters. The cell volume fraction in the beads ranged from 0.70 to 0.90, significantly higher than previously reported data. The effective diffusion coefficients conform to the random pore model. No diffusional interaction between acetophenone and phenethyl alcohol was observed. The addition of 2 vol% ethanol or methanol slightly increased the diffusivities. The thermodynamic partition coefficients were measured in the bead-free water-organic system and found to be an order of magnitude lower than the values calculated from the analysis of the diffusion data for the organic-bead system, suggesting that bead-free equilibrium data cannot be used in triphasic systems. (c) 1994 John Wiley & Sons, Inc.     

A comparison of two methods to assess apparent total tract digestibility of nutrients in dogs

ABSTRACT

The apparent total tract digestibility (ATTD) of nutrients can be assessed by total collection of faeces (TC), which is the reference method, or by the indicator method (IM). Little information is available on proper faecal sampling methodologies for IM in canines to obtain results comparable to TC. The objective of this study was to determine the minimum number of sub-samples required for the IM to make it comparable with TC. A total of 11 adult male dogs were individually housed in metabolism cages. Dogs had access to a grass yard to facilitate defaecation. Faecal sub-samples (1/day) were taken from the daily faecal output to prepare the pooled samples for IM, obtaining cumulative sample combinations of 3 (IM3), 4 (IM4), 5 (IM5), 6 (IM6) and 7 d (IM7). Digestibility of dry matter, gross energy, crude protein and crude fibre was similar between TC and IM5, IM6 and IM7 (p > 0.05). The IM7 presented the greatest statistical similarity with TC. Nevertheless, IM was not a good predictor of crude fibre digestibility. In conclusion, IM can replace the TC method in dogs to evaluate ATTD of several nutritional fractions as long as the composite sample is collected during seven consecutive days. For estimation of fibre digestibility by IM, longer collection periods are probably required.     

Cost‐effectiveness of Endoscopic Surveillance for Gastric Intestinal Metaplasia

Background: Patients with intestinal metaplasia (IM) are at increased risk for gastric cancer. Endoscopic surveillance has been shown to anticipate cancer diagnosis in an earlier stage. Cost‐effectiveness of endoscopic surveillance in IM patients is unknown. To assess the efficacy and cost‐effectiveness of an yearly endoscopic surveillance in patients with IM. Methods: A decision analysis model was constructed in order to compare a strategy of performing an EGD every year for a 10‐year period (surveillance strategy) following a new diagnosis of IM to a policy of nonsurveillance in a simulated cohort of 10,000 American patients. A 1.8% 10‐year cumulative incidence of gastric cancer in IM patients was estimated from the literature. Endoscopic surveillance was simulated to downstage the detected cancers by 58–84%. Costs of EGD and cancer care were estimated from Medicare reimbursement data. The main outcome measurement was the incremental cost‐effectiveness ratio. Results: The number of EGDs required to detect one cancer and to prevent one gastric cancer‐related death in the surveillance arm were 556 and 3738, respectively. The incremental cost‐effectiveness ratio of endoscopic surveillance as compared to a nonsurveillance policy was $72,519 per life‐year gained (5–95% percentiles Monte Carlo analysis: $54,843–$98,853). At sensitivity analysis, cancer incidence and the rate of downstaging were the most important variables. Conclusions: According to our simulation, the relatively high risk of cancer in patients with IM and the substantial efficacy of endoscopic surveillance in reducing cancer‐related mortality would support the cost‐effectiveness of an endoscopic surveillance program in patients with IM. Further research is needed before implementing it in the clinical practice.     

Growth of total fat and lean and of primal cuts is affected by the sex type

Knowledge of tissue and cuts growth depending on the sex could be used to improve performance and efficiency. Computed tomography (CT) is a non-invasive technology that enables the study of the body composition of live animals during growth. The aims of the present study were (1) to evaluate variation in the body composition of four sex types (SEX) of pigs (castrated males (CM), immunocastrated males (IM), entire males (EM) and females (FE)) at the live weight of 30, 70, 100 and 120 kg, assessed using CT; (2) to model the growth of the main tissues and cuts; and (3) to predict the mature BW (MBW) of the four SEX and establish the relationships between the growth models and the MBW. There were significant phenotypic differences in the allometric growth of fat and lean among SEX. For the lean tissue, FE and EM showed higher values of the b coefficient than CM and IM (1.07 and 1.07 v. 1.00 and 1.02, respectively) all of them close to unity, indicating a proportional growth rate similar to live weight and that this tissue developed faster in FE and EM than in CM and IM. However, these differences were not related to differences in estimated MBW. There were significant differences in estimated MBW among SEX, being higher in IM and EM than in CM and FE (303 and 247 v. 219 and 216 kg), however, the MBW may have been overestimated, especially for the IM. The poorer accuracy of the MBW estimate for the IM could be due to a maximum live weight of 120 kg in the experiment, or to the fact that this particular SEX presented two clear behaviours, being more similar to EM from birth to the second injection of the vaccine (130 days) and comparable with CM from that point to the final BW.     

The effects of grain treatment, grain feed level and grass silage feed value on the performance of and meat quality from, finishing beef cattle

A completely randomised design study involving 132 continental crossbred beef steers was undertaken to evaluate the effects of method of grain treatment and feed level, and grass silage feed value on animal performance, carcass characteristics and meat quality of beef cattle. Winter wheat was harvested and the grain was stored either ensiled crimped and treated with 4.5 l/t of a proprietary acid-based additive (crimped), ensiled whole and treated with 20 kg feed-grade urea per t (urea) or stored conventionally in an open bin treated with 3 l propionic acid per t. Two grass silages, of contrasting feed value (L and H) were ensiled. For the conventional, crimped and urea treatments, grain dry matter (DM) concentrations were 802, 658 and 640 g/kg, respectively. For the L- and H-feed value silages, DM concentrations were 192 and 240 g/kg and D values were 671 and 730 g/kg DM, respectively. The silages were offered as the sole forage supplemented with either conventional, crimped or urea-treated grain-based concentrate at either 3.5 or 6.0 kg DM per steer per day. The grain supplement consisted of 850 and 150 g/kg DM of grain and citrus pulp, respectively. For the conventional, urea and crimped treatments, DM intakes were 8.85, 9.43 and 9.04 kg/day (standard error (s.e.) = 0.129); estimated carcass gains were 0.60, 0.55 and 0.61 kg/day (s.e. = 0.020), respectively. For the low- and high- feed value grass silages, estimated carcass gains were 0.56 and 0.61 kg/day (s.e. = 0.014), respectively. For the low and high grain feed levels, estimated carcass gains were 0.56 and 0.61 kg/day, respectively. Grain treatment, grain feed level or silage feed value did not alter (P > 0.05) meat quality, lean colour or fat colour. There were significant silage feed value × grain feed level interactions (P < 0.05) for final live weight (LW) and daily live-weight gain (DLWG). Increasing grain feed level increased final LW and DLWG when offered with the low-feed value silage, however, grain feed level had no effect on final LW or DLWG when offered with the high-feed value silage. It is concluded that urea treatment of grain increased silage intake and feed conversion ratio (kg DM intake per kg carcass) and tended to decrease carcass gain. Crimping provides a biologically equally effective method to store grain as conventional methods. Improving grass silage feed value had a greater impact on animal performance than increasing grain feed level by 2.4 kg DM per day.     

A simple correlation for predicting effective diffusivities in immobilized cell systems

A simple correlation method has been developed to predict effective diffusivities of small molecules in heterogeneous materials such as immobilized cell systems. This correlation uses a single diffusivity measurement at one cell volume fraction to predict diffusivities for any other volume fraction of cell. The method has been applied to 20 sets of published diffusivity measurements in immobilized cell systems and accurately predicts affective diffusivities of molecules for the full range of cell fractions. It may also be used to predict effective diffusivities in heterogeneous materials in which the diffusivity of a molecule in each phase and the volume fraction of each phase are known. (c) 1996 John Wiley & Sons, Inc.     

Cytokines in the muscle tissue of idiopathic inflammatory myopathies: implications for immunopathogenesis and therapy

The inflammatory myopathies (IM), dermatomyositis (DM), polymyositis (PM) and idiopathic inclusion body myositis (IBM) are acquired immune-mediated myopathies. About their pathogenesis and etiology no definitive insights are available yet. Here we present a review of cytokine studies in IM. Combined with cellular immunohistochemical findings a model is presented describing a common mechanism of immune activation in IM. This model is based on a "hit" triggering local cytokine production with dominance of pro-inflammatory cytokines, like IFN-gamma and Th1-mediated activities. The altered Th1-Th2 balance necessitates detection of the anti-inflammatory arm of immune activation, which includes Th2-derived IL-4, IL-1, and Th3/Tr1 derived IL-10 and TGF-beta. Redirection of the ratio provides targets for novel immunotherapy by direct inhibition of the IFN-gamma-mediated Th1 response, stimulation of Th3/Tr1, or IL-4-secreting Th2-cells, negative feedback inhibition with IFN-beta and IFN-gamma and inactivation of MHC molecules.     

Nuclear volume in type I gastric intestinal metaplasia

OBJECTIVE: To retrospectively compare nuclear size of epithelial cells in type I intestinal metaplasia (IM) from various pathologic lesions of gastric mucosa. STUDY DESIGN: Endoscopic mucosal biopsies with type I IM from intestinal type gastric carcinoma (n = 25), chronic gastritis (n = 40) and benign ulcer (n = 32) cases were analyzed. After standard fixation, embedding, sectioning, routine hematoxylin and eosin and alcian blue--periodic acid--Schiff reaction (pH 1.0 and 2.5) staining were used for identification of IM. The mean point sampled nuclear intercept was estimated by the original test system and 100x objective at a total magnification of 1,200x. To obtain the mean nuclear volume, the cubed nuclear intercept was multiplied by pi/3. In each case 100 epithelial cell nuclei were analyzed. RESULTS: In type I IM in gastric carcinoma cases there was significantly greater nuclear volume (118.34 +/- 10.32 micron 3) than in type I IM in other pathologic states of gastric mucosa (77.72 +/- 8.58 micron 3). CONCLUSION: The karyometric findings of the present study suggest a difference between type I IM found in benign pathologic states and type I IM found in gastric mucosa surrounding carcinoma, despite morphologic and histochemical similarities. Nuclear volume may be used in early detection of precancerous states of gastric mucosa.     

家族高发性2型糖尿病的遗传模式研究

对1999～2000年门诊及住院的家族高发性2型糖尿病患者为先证者的136个大家系进行研究,以探讨该病的遗传模式。对家系人群采用Falconer 法估算遗传率,用Penrose法进行多基因分析,并用S.A.G.E-REGD软件拟合A型回归Logistic模型进行复合分离分析的方法,对家族高发性2型糖尿病家系进行研究。结果表明,136个大家系的2型糖尿病遗传率为94.07%±5.84%,提示在这些家系中可能有显性主基因存在。多基因分析研究表明,在该人群中,2型糖尿病因性别不同而存在两种遗传模式。复合分离分析拒绝单纯环境模型、非传递模型、共显性模型,接受隐性模型和显性模型,但隐性模型为最佳遗传模式。因此,2型糖尿病具有高度的遗传性和遗传异质性,总体表现为多因子遗传,在部分遗传背景较一致的家系人群中可能存在由主要基因决定的常染色体显性遗传。 Abstract:This study is to explore the genetic model of type 2 diabetes mellitus (type 2 DM) among the hereditary family.One hundred and thirty-six pedigrees of familial type 2 DM were studied.The heritability of type 2 DM was estimated according to Falconer's method and the multi-factorial inheritance analyzed according to Penrose's method.Complex segregation analysis was performed using S.A.G.E-REGD.The heritability of familial type 2 DM was 9407%±5.84%.Dominant major gene might influence the genesis of type 2 DM.Analysis of multi-factorial inheritance indicated that there be two genetic patterns respectively in male and female populations.By complex segregation analysis,environment,non-transmitted and co-dominant inheritance were rejected.Autosomal dominant (AD) inheritance and autosomal recessive (AR) inheritance was accepted but AR inheritance was the best pattern.This study suggested that type 2 DM had significant heritability and genetic heterogeneity,which appeared to be a disease of multi-factorial inheritance generally and autosomal dominant (AD) inheritance in part of pedigrees.     

Cardiovascular Variability Analysis and Baroreflex Estimation in Patients with Type 2 Diabetes in Absence of Any Manifest Neuropathy

Introduction

Indexes derived from spontaneous heart period (HP) and systolic arterial pressure (SAP) fluctuations can detect autonomic dysfunction in individuals with type 2 diabetes mellitus (DM) associated to cardiovascular autonomic neuropathy (CAN) or other neuropathies. It is unknown whether HP and SAP variability indexes are sensitive enough to detect the autonomic dysfunction in DM patients without CAN and other neuropathies.

Methods

We evaluated 68 males aged between 40 and 65 years. The group was composed by DM type 2 DM with no manifest neuropathy (n = 34) and healthy (H) subjects (n = 34). The protocol consisted of 15 minutes of recording of HP and SAP variabilities at rest in supine position (REST) and after active standing (STAND). The HP power in the high frequency band (HF, from 0.15 to 0.5 Hz), the SAP power in the low frequency band (LF, from 0.04 to 0.15 Hz) and BRS estimated via spectral approach and sequence method were computed.

Results

The HF power of HP was lower in DM patients than in H subjects, while the two groups exhibited comparable HF power of HP during STAND. The LF power of SAP was similar in DM and H groups at REST and increased during STAND in both groups. BRSs estimated in the HF band and via baroreflex sequence method were lower in DM than in H and they decreased further during STAND in both populations.

Conclusion

Results suggest that vagal control of heart rate and cardiac baroreflex control was impaired in type 2 DM, while sympathetic control directed to vessels, sympathetic and baroreflex response to STAND were preserved. Cardiovascular variability indexes are sensitive enough to typify the early, peculiar signs of autonomic dysfunction in type-2 DM patients well before CAN becomes manifest.     

A study of morphological and genetic distance among four Indian villages of Nicaragua

The degree of biological difference between four native populations from Nicaragua has been sought by measuring their morphological distance (DM) and genetic distance (DG). The calculation methods proposed by Sanghvi (1953) were applied. DM is estimated by taking into account first a set of craniometric measurements DM (C), then a set of somatometric measurements DM (S), and finally by calculating both sets jointly DM (CS). Comparison is also carried out by measuring DM according to a set of 13 indices (7 cranial and 6 somatic ones) DM (I). The position acquired by each population with respect to the others differs according to its determination as DM (C) or DM (S). On the contrary, DM (CS) and DM (I) seem to yield identical results, though little comparable with those obtained by comparison carried out according to the traditional anthropological methods. Moreover, the position of the four populations for DM at various levels in no case corresponds to that obtained by the measurement of DG.     

Action of chondroitinases. II. Numerical calculation of the degree of multiple attack.

Further investigation was carried out on the action patterns of two chondroitinase-AC [EC 4.2.2.5.] preparations obtained from Arthrobacter aurescens and Flavobacterium heparinum. To infer the action patterns of the chondroitinases, we proposed a new method for the calculation of the degree of multiple attack, based on the concept established by Robyt and French ((1967) Arch. Biochem. Biophys. 122, 8-16). It was shown that the degree of multiple attack (DM) is represented by the ratio of the initial velocity of number-average degree of scission to that of viscosity-average degree of scission. By this method, DM for A-Chase was estimated to be 3.03 and for F-chase, 1.31.     

In situ intestinal digestibility of dry matter and crude protein of cereal grains and rapeseed in sheep

The ruminal degradation and intestinal digestibility (ID) of dry matter (DM) and crude protein (CP) of different feed samples were measured in two trials by using nylon bag and rumen outflow rate techniques in three wethers cannulated in the rumen and in the duodenum. In trial 1, three samples of grains of wheat, barley, and corn treated by cooking (TW, TB, and TC, respectively) were studied together with a sample of untreated corn grains (CG) of different origin. In trial 2, these studies were carried out on a sample of rapeseed (RS) and on a mix of this same sample and rapeseed meal (in proportions 70:30) treated by cooking (TR). In both trials, the animals were fed at the same intake level (40 g DM x kg(-1) LW0.75) with 2:1 (DM basis) forage to concentrate diets. Rumen degradation rates of DM were high in the treated cereals (between 11.0 and 14.2% x h(-1)) and low in the CG (6.35% x h(-1)), whereas for CP these rates were low in all cereals. For DM, in all cereals, ID decreased linearly as the ruminal incubation time increased. The values of intestinal effective digestibility (IED), calculated from these functions and from the rumen outflow, were respectively: 86.4, 62.1, 51.5, and 67.9%. For CP, ID was unaffected by the ruminal incubation time in corn samples, whereas in TW and TB a reduction of these values was only observed for the time of 48 h. The values of IED of CP for CG, TW, TB and TC were: 82.6, 88.9,82.5, and 91.6%, respectively. Rumen degradation rates of the RS and TR samples were 8.35 and 8.23% x h(-1) for DM and 12.0 and 9.59% x h(-1) for CP. In RS, the ID of DM and CP showed a downward trend with an increase of the ruminal incubation time, as modelled according to an exponential function. This same trend was observed for TR after a lag period estimated at 7.53 and 6.51 h for DM and CP, respectively. The values of IED of RS and TR were respectively 56.5 and 50.8% for DM and 71.9 and 80.1% for CP. These same results were also determined by a simplified method using a sample pooled to be representative of the rumen outflow of undegraded feed. The respective values for RS and TR were 54.8 and 51.6 for DM and 65.8 and 78.9% for CP. This method seems to be a promising technique to estimate IED, although more studies are needed to improve its accuracy.     

Contribution of root cap mucilage and presence of an intact root cap in maize (Zea mays) to the reduction of soil mechanical impedance

BACKGROUND AND AIMS: The impedance to root growth imposed by soil can be decreased by both mucilage secretion and the sloughing of border cells from the root cap. The aim of this study is to quantify the contribution of these two factors for maize root growth in compact soil. METHODS: These effects were evaluated by assessing growth after removing both mucilage (treatment I -- intact) and the root cap (treatment D -- decapped) from the root tip, and then by adding back 2 micro L of mucilage to both intact (treatment IM -- intact plus mucilage) and decapped (treatment DM -- decapped plus mucilage) roots. Roots were grown in either loose (0.9 Mg m(-3)) or compact (1.5 Mg m(-3)) loamy sand soils. Also examined were the effects of decapping on root penetration resistance at three soil bulk densities (1.3, 1.4 and 1.5 Mg m(-3)). KEY RESULTS: In treatment I, mucilage was visible 12 h after transplanting to the compact soil. The decapping and mucilage treatments affected neither the root elongation nor the root widening rates when the plants were grown in loose soil for 12 h. Root growth pressures of seminal axes in D, DM, I and IM treatments were 0.328, 0.288, 0.272 and 0.222 MPa, respectively, when the roots were grown in compact soil (1.5 Mg m(-3) density; 1.59 MPa penetrometer resistance). CONCLUSIONS: The contributions of mucilage and presence of the intact root cap without mucilage to the lubricating effect of root cap (percentage decrease in root penetration resistance caused by decapping) were 43 % and 58 %, respectively. The lubricating effect of the root cap was about 30 % and unaffected by the degree of soil compaction (for penetrometer resistances of 0.52, 1.20 and 1.59 MPa).     

L-glutamate enhances the expression of Thermus maltogenic amylase in Escherichia coli

Escherichia coli BL21 (DE3) transformed with a thermostable Thermus maltogenic amylase (ThMA), isolated from a Gram-negative bacterium Thermus strain IM6501, grew well and efficiently produced ThMA in a complex medium but not in a chemically defined medium (DM). By supplementing L-glutamate to DM medium, both the specific growth rate and ThMA expression significantly increased. Alterations in the cellular responses of recombinant E. coli to L-glutamate were analyzed at the protein level by two-dimensional gel electrophoresis and mass spectrometry. The ppGpp synthase (RelA) was significantly reduced in cells grown with L-glutamate and was consistent with the low level of ppGpp, an indicator of stringent response. On the other hand, protein chain elongation factor (EF-Tu) and manganese-containing superoxide dismutase (MnSOD), which protects cells against oxidative damage, was significantly elevated by L-glutamate supplementation. These results indicate that L-glutamate enhances ThMA expression and increases the E. coli growth rate not only by overcoming the stringent response but also by increasing the synthesis of EF-Tu and MnSOD.     

Method development and validation for the simultaneous determination of imatinib mesylate and N-desmethyl imatinib using rapid resolution high performance liquid chromatography coupled with UV-detection

We developed a simple and sensitive method for the simultaneous detection of imatinib mesylate (IM) and its active metabolite, N-desmethyl imatinib (M1), in human serum samples. Separation was successfully achieved using an Agilent(?) ZORBAX Eclipse plus C(18) reversed phase column (50 mm × 2.1 mm, i.d.; 1.8 μm) under isocratic mobile phase conditions consisting of acetonitrile: 0.02 M potassium dihydrogen phosphate with 0.2% triethylamine at pH 3 (25:75, v/v) and ultra-violet detection was achieved at 235 nm. Extraction of the target compounds was completed using 100% cold acetonitrile. Good linearities (r(2)>0.99) for both IM and M1 were achieved for the concentration ranges of 50-1800 ng/mL and 50-360 ng/mL, respectively. The detection limits were 20 ng/mL and 10 ng/mL for M1 and IM, respectively. The intra- and inter-day precisions were less than 1% with percent recoveries of more than 90%. The method was successfully applied to calculate the pharmacokinetic parameters of chronic myeloid leukemia patients receiving imatinib. The method is suitable to be routinely applied for determination of IM and M1 in serum.