A randomized,double‐blind,placebo controlled,multi‐center study of intravenous iron sucrose and placebo in the treatment of restless legs syndrome

Iron deficiency may exacerbate symptoms in the Restless Legs Syndrome (RLS). We investigated the effect of intravenous iron sucrose or placebo on symptoms in patients with RLS and mild to moderate iron deficit. Sixty patients with primary RLS (seven males, age 46 (9) years, S‐ferritin ≤45 μg/L) recruited from a cohort of 231 patients were randomly assigned in a 12‐months double‐blind, multi‐centre study of iron sucrose 1000 mg (n = 29) or saline (n = 31). The primary efficacy variable was the RLS severity scale (IRLS) score at week 11. Median IRLS score decreased from 24 to 7 (week 11) after iron sucrose and from 26 to 17 after placebo (P = 0.123, N.S. for between treatment comparison). The corresponding scores at week 7 were 12 and 20 in the two groups (P = 0.017). Drop out rate because of lack of efficacy at 12 months was 19/31 after placebo and 5/29 patients after iron sucrose (Kaplan–Meier estimate, log rank test P = 0.0006) suggesting an iron induced superior long term RLS symptom control. Iron sucrose was well tolerated. This study showed a lack of superiority of iron sucrose at 11 weeks but found evidence that iron sucrose reduced RLS symptoms both in the acute phase (7 weeks) and during long‐term follow up in patients with variable degree of iron deficiency. Further studies on target patient groups, dosing and dosing intervals are warranted before iron sucrose could be considered for treatment of iron deficient patients with RLS. © 2009 Movement Disorder Society     

Chromosome mapping ofSoa,a gene influencing gustatory sensitivity to sucrose octaacetate in mice

Strain distribution patterns among recombinant inbred strains suggested that a locus influencing taste sensitivity to sucrose octaacetate was on chromosome 6. A location forSoa was established by linkage analysis of behavioral and electrophoretic data from outbred and congenic strains and from test-cross progeny. Haplotyping of 41 outbred CFW-Cr animals with a cDNA probe showed perfect cosegregation ofSoa andPrp, a gene for salivary proline-rich proteins. Five of twelve B6. SW-Soa ^a strains were found to retainLdr-1, lactate dehydrogenase regulator-1, on chromosome 6 as an allelic passenger from the SWR/J donor strain (source of theSoa ^a Taster allele). Centimorgan distance was estimated using the ABP/Le linkage-testing strain (non-Taster,Soa ^b) and the SWR/J strain (Taster,Soa ^a) in a testcross breeding system. The data are consistent with a position for theSoa locus on mouse chromosome 6, 62 cM from the centromere.This research was supported in part by Grants DC00150 (G. W.) and DE003658 (E.A.A.). This paper is based on a thesis submitted to the Florida State University by the first author in partial fulfillment of the requirements for the Master of Sciences degree.     

Isolation and partial characterisation of neuronal growth cones from neonatal rat forebrain

We have devised a method for the isolation of viable neuronal growth cones from neonatal rat forebrain. The method involves differential and density gradient centrifugation and exploits the relatively low buoyant density (approximately 1.018 g/cm3) of growth cones. There are no known biochemical markers for growth cones and it was necessary therefore to monitor for their presence during the isolation using transmission electron microscopy. Several criteria were used to identify isolated growth cones including the presence of filopodia, an extensive system of branching, tubular smooth endoplasmic reticulum and a region rich in microfilaments subjacent to the plasma membrane. These morphological features are similar to those of growth cones identified unequivocally in intact developing brain and in tissue culture. Electron microscopical analysis showed that greater than 90% of membrane-bound, identifiable objects in one fraction were growth cones by these criteria. The major contaminant consisted of membrane sacs and vesicles of unidentified origin. There were only small amounts of isolated rough endoplasmic reticulum and mitochondria. Isolated growth cones were roughly spherical in shape with a diameter of 1.9 +/- 0.5 micron (mean +/- 1 SD). They usually contained mitochondria, large granular vesicles and small vesicles, and occasionally contained coated vesicles, lysosomes, lamellar bodies and multivesicular bodies, and only very rarely, intermediate filaments. Occasionally, growth cones had rudimentary synapses on them. The viability of isolated growth cones was investigated by observing their behaviour in short-term culture. After a few hours in culture on poly-D-lysine-coated coverslips, growth cones flattened down and extended filopodia-like processes. This behaviour was inhibited by cytochalasin B and reversibly by cold (4 degrees C). We conclude that physiologically active growth cones can be isolated rapidly and in large numbers by the method described here.     

中药配方颗粒辅料麦芽糊精的HPLC定量检测与中红外光谱半定量检测

目的 建立中药配方颗粒辅料麦芽糊精的检测方法,为中药配方颗粒质量评价提供分析技术支持。方法 建立配方颗粒中果糖、葡萄糖、蔗糖、麦芽糖、乳糖的HPLC定量检测方法,根据样品经糖化酶水解后的葡萄糖增加量换算出麦芽糊精含量。基于中药浸膏与麦芽糊精不同比例混合物的中红外光谱（mid-infrared spectroscopy,MIRS）特征峰差异,建立配方颗粒中麦芽糊精的半定量检测方法。结果 所建立的HPLC方法可以准确定量检测配方颗粒中果糖、葡萄糖、蔗糖、麦芽糖、乳糖。如果中药浸膏自身含有较多可经糖化酶水解产生葡萄糖的成分（淀粉、蔗糖等）,HPLC检测配方颗粒中麦芽糊精时存在系统性正误差,可通过中药浸膏酶解后葡萄糖增加量、待测样品酶解后蔗糖减小量等部分修正正误差。如果中药浸膏自身少含或不含淀粉、蔗糖等可经糖化酶水解产生葡萄糖的成分,HPLC检测配方颗粒中麦芽糊精时存在系统性负误差,换算公式偏差、麦芽糊精纯度、样品处理损失等导致根据样品酶解后葡萄糖增加量计算的麦芽糊精“检测含量”预期低于根据生产投料计算的麦芽糊精“名义含量”。麦芽糊精含量越高,配方颗粒MIRS中1200～900 cm-1区域的...     

基于指纹图谱和多指标含量测定的蜜远志质量评价

目的 建立指纹图谱和多指标定量的蜜远志质量评价方法。方法 Kromasil 100-5-C₁₈色谱柱（250 mm×4.6 mm,5 μm）为固定相,以乙腈（A）-0.2%磷酸水溶液（B）为流动相进行梯度洗脱,体积流量为1.0 mL/min,检测波长318 nm。对16批蜜远志建立指纹图谱,确定共有峰并进行相似度分析结合化学计量学分析。采用Waters Xbridge Shiled RP C₁₈色谱柱（250 mm×4.6 mm,5 μm）进行同时测定远志（口山）酮III、3,6''-二芥子酰基蔗糖、细叶远志皂苷、远志酸、远志皂苷元的含量。结果 在指纹图谱研究中,标定了27个共有峰,结合对照品和HPLC-Q-TOF-MS共指认8个成分,分别为西伯利亚远志糖A5、西伯利亚远志糖A6、远志（口山）酮IX、远志（口山）酮III、3,6''-二芥子酰基蔗糖、细叶远志皂苷、远志酸、远志皂苷元。聚类分析将16批样品分为3类;经主成分分析,主成分1～3是影响蜜远志质量评价的主要因子。多指标含量测定中远志（口山）酮III、3,6''-二芥子酰基蔗糖、细叶远志皂苷、远志酸、远志皂苷元的质量分数分别为0.027%～0.068%、0.074%～0.798%、1.1%～1.4%、0.15%～0.36%、0.15%～0.37%。经方法学验证,线性关系良好（r≥0.999 6）,平均加样回收率为97.15%～100.9%。结论 所建立的指纹图谱结合多指标含量测定方法准确、高效,特征性强,可为蜜远志饮片的质量评价和临床应用提供有效参考。     

Some technical improvements for the voltage clamp with the double sucrose gap

Summary A new double sucrose gap chamber using electrodes of high stability and low resistance is described. The size of each sucrose gap and the central node can be modified independently so that this chamber is suitable for research on multicellular or unicellular excitable preparation under voltage clamp conditions. The shape of the central channel is made so as to reduce the series resistance between the preparation and the recording electrode to a minimum. A window made in the floor of the chamber allows to observe the preparation under a microscope.This work was performed under the I.N.S.E.R.M. grant no. 17.75.40.     

The Relationship Between Protein Aggregation and Molecular Mobility Below the Glass Transition Temperature of Lyophilized Formulations Containing a Monoclonal Antibody

Purpose. To find out if the physical instability of a lyophilized dosage form is related to molecular mobility below the glass transition temperature. Further, to explore if the stability data generated at temperatures below the glass transition temperature can be used to predict the stability of a lyophilized solid under recommended storage conditions. Methods. The temperature dependence of relaxation time constant, , was obtained for sucrose and trehalose formulations of the monoclonal antibody (5 mg protein/vial) from enthalpy relaxation studies using differential scanning calorimetry. The non-exponentiality parameter, , in the relaxation behavior was also obtained using dielectric relaxation spectroscopy. Results. For both sucrose and trehalose formulations, the variation in with temperature could be fitted Vogel-Tammann-Fulcher (VTF) equation. The two formulations exhibited difference sensitivities to temperature. Sucrose formulation was more fragile and exhibited a stronger non-Arrhenius behavior compared to trehalose formulation below glass transition. Both formulations exhibited <2% aggregation at t values <10, where t is the time of storage. Conclusions. Since the relaxation times for sucrose and trehalose formulations at 5°C are on the order of 10⁸ and 10⁶ hrs, it is likely that both formulations would undergo very little (<2%) aggregation in a practical time scale under refrigerated conditions.     

高糖摄入对大鼠糖和脂质代谢及纤溶活性影响

①目的　观察高糖摄入对正常 Ｓｐｒａｇｕｅ Ｄａｗ ｌｅｙ（ Ｓ Ｄ）大鼠糖及胰岛素代谢、脂质代谢和纤溶活性的影响。②方法　将雄性 Ｓ Ｄ 大鼠１６ 只随机分为２ 组,实验组于饮水中添加蔗糖（１２０ｇ／ Ｌ）喂养４２ｄ,对照组给以正常饮水,每７ｄ 定时测量体质量（ Ｂ Ｗ）１ 次,实验最后１ｄ 取血测空腹血糖（ Ｆ Ｂ Ｇ）,空腹血清胰岛素（ Ｉ Ｎ Ｓ）,甘油三酯（ Ｔ Ｇ）,总胆固醇（ Ｔ Ｃ）,高密度脂蛋白胆固醇（ Ｈ Ｄ Ｌ Ｃ）水平及血浆纤溶酶原激活物抑制物１（ Ｐ Ａ Ｉ１）活性等指标。③结果与对照组相比,实验组 Ｆ Ｂ Ｇ,血清 Ｔ Ｇ 水平及血浆 Ｐ Ａ Ｉ１ 活性均升高（ｔ＝ ２．３６～４．７３, Ｐ＜ ０．０５,０．０１）;血清 Ｔ Ｃ及 Ｈ Ｄ Ｌ Ｃ水平两组差异无显著性（ｔ＝ １．３７,０．８４, Ｐ＞ ０．０５）;两组血清 Ｉ Ｎ Ｓ水平虽然差异无显著性（ｔ＝ ０．７７, Ｐ＞０．０５）,但是经协方差分析纠正体质量因素后,实验组与对照组相比存在高胰岛素血症（ Ｆ＝ ５．７４, Ｐ＜ ０．０５）;直线相关分析提示,实验组血清 Ｉ Ｎ Ｓ水平与 Ｂ Ｗ ,血清 Ｔ Ｇ水平及血浆 Ｐ Ａ Ｉ１ 活性呈高度正相关,对照组血清 Ｉ Ｎ Ｓ水平与 Ｂ Ｗ 及     

蔗糖对红细胞膜脂流动性测定稳定性的影响

研究蔗糖对测定红细胞膜脂流动性稳定性的影响。应用荧光偏振技术测定膜脂流动性,并对比其在单纯悬浮液和含0．25mol／L蔗糖悬浮液中的测定结果的变异系数。结果在含有蔗糖悬浮液中测得结果的变异系数要比单纯悬浮液中的小3～5倍。结果表明,加入蔗糖对膜脂流动性的测定具有稳定性作用。     

A comparison between the effect of cholecystokinin octapeptide and apomorphine on ingestion of intraorally administered sucrose in male rats

The involvement of dopamine receptors in the inhibitory effect of Cholecystokinin octapeptide on ingestive behaviour was investigated. Male rats were infused intraorally with a 1 M solution of sucrose and the amount ingested after treatment with the dopamine receptor agonist apomorphine was compared with that after treatment with Cholecystokinin octapeptide. The test allows a distinction between the consummatory aspects of ingestive behaviour, i.e. responses used to ingest food, from the appetitive aspects, i.e. responses used to obtain food, because it ignores the latter aspects. Comparisons were also made between the effects of apomorphine and Cholecystokinin octapeptide on pellet intake, a test in which the rat has to display appetitive ingestive behaviour. Injection of apomorphine (400 μg) increased the concentration of plasma apomorphine within 0.3 min and the concentration of dopamine in the cerebrospinal fluid within 1 min of injection and induced behavioural stereotypes within 10 min in food-deprived male rats. Plasma apomorphine and cerebrospinal fluid dopamine levels had decreased by 30 min and the behavioural stereotypies had decreased by 40 min after the injection. Injection of apomorphine also inhibited the consumption of food pellets and the ingestion of sucrose. Inhibition of pellet and sucrose ingestion paralleled the effect of apomorphine on Stereotypie behaviour. Thus, injection of a dopamine receptor agonist is followed by alterations in plasma levels of the agonist, cerebrospinal fluid dopamine levels and in Stereotypie and ingestive behaviour which occur in parallel, in an inverted U-shaped manner and with a temporal delay between each event. These results show a close correlation between dopamine receptor stimulation and inhibition of ingestive behaviour. However, reversal of the inhibitory effect of apomorphine on ingestive behaviour required pretreatment with a lower dose of a dopamine receptor antagonist (cis-flupentixol) (0.1 mg) than reversal of Stereotypie behaviour (0.8 mg). The effect of dopamine receptor stimulation on consummatory ingestive behaviour is thus relatively weak and not secondary to the induction of Stereotypic behaviour. Treatment with a high dose of cis-flupentixol (0.8 mg) caused a prolonged period of immobility but had no effect on the ingestion of sucrose. Dopamine receptor blockade, therefore, interferes with appetitive, but not consummatory ingestive behaviour. Injection of Cholecystokinin octapeptide (5 μg) suppressed pellet and sucrose intake in a manner comparable to that of apomorphine, but induced no behavioural stereotypes and caused a gradual, rather than inverted U-shaped, increase in the concentration of dopamine in the cerebrospinal fluid that did not correlate with the effect on ingestive behaviour. Furthermore, while the inhibitory effect of apomorphine on the ingestion of sucrose was reversed by pretreatment with a low dose of cis-flupentixol (0.1 mg), the inhibitory effect of Cholecystokinin octapeptide was only partially reversed by cis-flupentixol and a higher dose (0.8 mg) was required. Blockade of cholecystokinin-A receptors, by treatment with L-364,718, but not cholecystokinin-B receptors, by treatment with L-365,260, blocked the inhibitory effect of Cholecystokinin octapeptide and, by itself, L-364,718 increased the amount of ingested sucrose. The inhibitory effect of Cholecystokinin octapeptide on consummatory ingestive behaviour, which is mediated by cholecystokinin-A receptors, is likely to involve mechanisms in addition to dopaminergic ones.