Abstract  Mass spectrometry has become an indispensable tool for the global study of metabolites (metabolomics), primarily using electrospray ionization mass spectrometry (ESI-MS). However, many important classes of molecules such as neutral lipids do not ionize well by ESI and go undetected. Chemical derivatization of metabolites can enhance ionization for increased sensitivity and metabolomic coverage. Here we describe the use of tris(2,4,6,-trimethoxyphenyl)phosphonium acetic acid (TMPP-AA) to improve liquid chromatography (LC)/ESI-MS detection of hydroxylated metabolites (i.e. lipids) from serum extracts. Cholesterol which is not normally detected from serum using ESI is observed with attomole sensitivity. This approach was applied to identify four endogenous lipids (hexadecanoyl-sn-glycerol, dihydrotachysterol, octadecanol, and alpha-tocopherol) from human serum. Overall, this approach extends the types of metabolites which can be detected using standard ESI-MS instrumentation and demonstrates the potential for targeted metabolomics analysis.

Abstract  After patch testing of several groups of patients we analyzed the development of contact allergy to wool wax alcohols and emulsifying fatty alcohols in the last years. For identifying the allergens we used high purified n-alcanols of the chain length of C-8 to C20, lanosterol and aliphatic diols. We found no reaction to cetyl and stearyl alcohol at all - the main allergens in literature. Most of the patch test reactions were caused by n-alcanols of C10H21OH to C15H31OH, but also by lanosterol and aliphatic diols. Alcohols of the chain length C-12 and C-14 we found in all gaschromatographically analyzed samples. Therefore we like to propose allergenfree emulgators for patients on risk.

Abstract  The MCF-7 cell line (human breast epithelial cells) accumulates fatty alcohols. The fatty alcohols were identified as C16:0, C18:0, and C18:1 alcohols by thin-layer chromatography and gas chromatography/mass spectrometry. This accumulation of alcohols in MCF-7 was found in cultures of MCF-7 cells obtained from other laboratories but not in a variety of unrelated cell lines. The presence of the alcohols suggested an aberrant ether lipid metabolism in the MCF-7 cells. Therefore, the capacity for either lipid biosynthesis was evaluated using cells incubated with either [14C]stearyl alcohol or [14C]stearic acid. MCF-7 cells incorporated less than 0.4% of the [14C]alcohol into ether-linked phospholipids, whereas the AB589 breast epithelial cells, used as a "normal control" for comparisons, did not accumulate fatty alcohol and incorporated approximately 20% of the radiolabeled alcohol into phospholipids containing ether linkages. Although the MCF-7 cells were unable to effectively incorporate the fatty alcohol into ether linkages, the cells were able to oxidize the alcohol to fatty acid. When incubated with [14C]stearic acid, the conversion to radiolabeled fatty alcohol in MCF-7 cells was approximately four times higher than the alcohol levels found in AB589 cells. While deficient in the ability to synthesize ether linkages, the MCF-7 cells did incorporate radiolabeled hexadecylglycerol, a precursor containing an ether linkage, into phospholipids. Collectively, the data indicate that the MCF-7 cells possess a deficiency in the alkyl DHAP synthase activity. A near absence of ether-linked lipids in the MCF-7 cells was indicated by the radiolabeling studies, and this finding was corroborated by results from HPLC analysis. Analyses of the partial glycerides, obtained from the enzymatic hydrolysis of cellular phospholipids, found only trace levels of ether lipids in the MCF-7 cells. The aberration in ether lipid biosynthesis did not correlate with the expression of the multidrug resistance phenotype in a series multidrug resistant MCF-7 variants. The results are discussed relative to the use of the MCF-7 cells as a model for investigations of ether lipid biosynthesis and the cellular physiology of ether lipids.

Abstract  Bladder dysfunction secondary to benign prostatic hyperplasia (BPH) is a major affliction of aging men. BPH can initiate both irritative lower urinary tract symptoms (urgency, frequency, and nocturia), and obstructive symptoms (reduced flow rate, increased micturition pressure, increased duration of micturition, and incomplete emptying). Although these symptoms are related to the effect of the enlarging prostate and subsequent urethral obstruction, there appears to be no direct relationship between prostate size or composition and severity of symptoms.3,13,17,19,23 In addition, the results of standard urodynamic evaluations do not correlate well with the severity of patient symptoms; standard urodynamics cannot accurately predict either level of bladder pathology or potential for recovery following surgery.3,13,17,19,23 One major problem is that current methods of urodynamic analysis, including pressure measurements, are indirect assessments of detrusor power or contractile function.

Abstract  A new oxyneolignan (rel-(7 alpha,8 beta)-3-methoxy-4',7-epoxy-8,3'-oxyneolignan-4,9,9'-triol) with a scarce C(7)-O-C(4') and C(8)-O-C(3') epoxy linkage, along with eight known compounds, abieta-8,11,13-triene, sandaracopimara-8(14), 15-diene, 6,7-dehydroferruginol methyl ether, 18-hydroxydehydroabietane, sandaracopimara-8(14), 15-dien-18-ol, docosanol, sugiol and palmitic acid, were isolated from the chloroform-soluble fraction of the acetone extract from Juniperus brevifolia leaves. Their structures were established on the basis of the spectral evidences and direct comparison with values from literature data. (C) 2010 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.

Abstract  The ATPase activity of the (Ca2+-Mg2+)-ATPase purified from skeletal muscle sarcoplasmic reticulum and reconstituted into phosphatidylcholine bilayers of defined composition depends on the fatty acyl chain length of the surrounding phospholipid. The stoichiometry of Ca2+ binding to the ATPase is also sensitive to fatty acyl chain length, changing from the normal two Ca2+ ions bound per ATPase molecule to one Ca2+ ion bound for the ATPase reconstituted with phosphatidylcholines of chain lengths C12, C14, or C24. For the ATPase reconstituted with mixtures of phosphatidylcholines where one phosphatidylcholine supports a Ca2+ binding stoichiometry of two and the other a stoichiometry of one, a highly cooperative change in binding stoichiometry with change in phospholipid composition is observed, suggesting that the effects of phospholipids follow from binding to a large number of sites at the lipid-protein interface of the ATPase. For the ATPase reconstituted with either 1-myristoyl-2-oleoylphosphatidylcholine or 1-oleoyl-2-myristoylphosphatidylcholine, the stoichiometry of Ca2+ binding is the normal two per ATPase molecule. Effects of short-chain phosphatidylcholines on Ca2+ binding stoichiometry and on ATPase activity can be reversed by addition of androstenol, oleic acid, methyl oleate, or oleyl alcohol but these molecules have no effect on the ATPase reconstituted with dinervonylphosphatidylcholine (C24:1). For the ATPase reconstituted with phosphatidylcholines with chain lengths between C16 and C22, release of the two bound Ca2+ ions is sequential, with release of the second Ca2+ being inhibited by high concentrations of Ca2+ in the bathing medium. For the ATPase reconstituted with phosphatidylcholines of chain lengths C14 or C24, release of the single bound Ca2+ is only slightly inhibited by the presence of Ca2+ in the medium. For the ATPase reconstituted with phosphatidylcholines of chain lengths between C16 and C24, removal of bound Ca2+ results in a decrease in tryptophan fluorescence intensity, whereas for the ATPase reconstituted with phosphatidylcholines of chain lengths C12 or C14, removal of bound Ca2+ results in an increase in tryptophan fluorescence intensity. In mixtures of phosphatidylcholines, changes in the tryptophan response mirror changes in Ca2+ binding stoichiometry.

Abstract  Mitotic checkpoint impairment is present in human lung cancers with chromosomal instability (CIN). Spindle-checkpoint genes have been reported to be mutated in several human cancers; but these mutations are infrequent. Recent reports suggest that the hBUBR1 gene may play an important role in mitotic checkpoint control and in mitotic checkpoint impairment in human cancers. We analyzed the expression of hBUBR1 in lung cancer cell lines using real time quantitative RT-PCR. The expression of BUBR1 was found to be up-regulated in all of these cell lines. In addition, we cloned and characterized the promotor region of hBUBR1 and determined its genomic structure, which includes 23 exons. The open reading frame (ORF) of the hBUBR1 gene comprises exons 1 through 23. There are GC-rich regions located at the flanking region and about 150 bp upstream from exon 1. The promoter region (424 bp upstream from exon 1) showed promoter activity and includes multiple transcription factor consensus binding motifs, including those for Sp1, Nkx-2, CdxA, SRY, MyoD, Ik-2, HNF-3b, Staf, Oct-1, Nkx-2, v-Myb, and AML 1a. Multiple pathways leading to activation of those binding factors may contribute to hBUBR1 gene transcription. Knowledge of the genomic structure and the promoter region of the hBUBR1 gene will facilitate investigation of its role in mitotic checkpoint control and tumor progression in human cancers. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.

Abstract  The autumn gum moth, Mnesampela privata (Guenee) (Lepidoptera: Geometridae), is native to Australia and can be a pest of plantation eucalypts. Field-collected and laboratory-reared female autumn gum moths were dissected to remove glands likely to contain components of the sex pheromone. Using gas chromatography (GC) and combined gas chromatography-mass spectrometry (GC-MS), three compounds were identified from female extracts, namely (3Z,6Z,9Z)-3,6,9-nonadecatriene, 1-hexadecanol and 1-octadecanol (confirmed by comparison with synthetic samples). Nonadecatriene elicited an antennal response in male autumn gum moth during gas chromatographic analyses combined with electroantennographic detection (GC-EAD). In electroantennogram (EAG) recording male M. privata antennae responded to the nonadecatriene. Nonadecatriene was synthesised via Kolbe electrolysis, starting with (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid (linolenic acid) and propanoic acid or via an alternative four-step method also starting from linolenic acid. In field trials (3Z,6Z,9Z)-3,6,9-nonadecatriene proved attractive to male moths. Thus, we conclude that (3Z,6Z,9Z)-3,6,9-nonadecatriene is a sex pheromone component of autumn gum moth. This component has been identified in extracts from other geometrids in the same subfamily, Ennominae. However, to our knowledge this is the first example where (3Z,6Z,9Z)-3,6,9-nonadecatriene has been found in females and also proved attractive to male moths when presented on its own. Our results are discussed in relation to other geometrid pheromones.

Abstract  Sjogren-Larsson syndrome (SLS) is an inherited disorder associated with deficient oxidation of long-chain aliphatic alcohols. Previous studies have reported modest elevations in total (free + esterified) fatty alcohols in SLS, but free fatty alcohols have not been selectively measured, in part because of their low concentrations in most tissues and the presence of trace fatty alcohol contaminants in some solvents used for their analysis. We adapted methods to measure free fatty alcohols in cultured cells and plasma that minimize exogenous alcohol contamination. Fatty alcohols were analyzed as acetate derivatives, using capillary column gas chromatography. By this method, cultured skin fibroblasts from SLS patients were found to have 7- and 8-fold elevations in the mean content of hexadecanol (16:0-OH) and octadecanol (18:0-OH), respectively. The mean plasma 16:0-OH and 18:0-OH concentrations in SLS patients (n = 11) were 9- and 22-fold higher than in normal controls, respectively. In SLS fibroblasts, most of the fatty alcohol (59%) that accumulated was free rather than esterified alcohol, whereas free alcohol accounted for 23% of the total alcohol in normal cells. These results indicate that elevations in free fatty alcohols provide a sensitive marker for the enzymatic defect in SLS. The ability to measure free fatty alcohols in cultured cells and plasma should prove useful for investigations of normal fatty alcohol metabolism and the deranged metabolism in SLS.

Abstract  Human prostate was used as a source of 5 alpha reductase. Compounds were incubated with an enzyme preparation and [3H]testosterone. [3H]-dihydrotestosterone production was measured to calculate 5 alpha reductase activity. IC50 values (ng/ml) were finasteride = 1; Permixon = 5,600; Talso = 7,000; Strogen Forte = 31,000; Prostagutt = 40,000; and Tadenan = 63,000. Bazoton and Harzol had no activity at concentrations up to 500,000 ng/ml. In castrate rats stimulated with testosterone (T) or dihydrotestosterone (DHT), finasteride, but not Permixon or Bazoton, inhibited T stimulated prostate growth, while none of the three compounds inhibited DHT stimulated growth. These results demonstrate that finasteride inhibits 5 alpha reductase, while Permixon and Bazoton have neither anti-androgen nor 5 alpha reductase inhibitory activity. In addition, in a 7 day human clinical trial, finasteride, but not Permixon or placebo, decreased serum DHT in men, further confirming the lack of 5 alpha reductase inhibition by Permixon. Finasteride and the plant extracts listed above do not inhibit the binding of DHT to the rat prostatic androgen receptor (concentrations to 100 micrograms/ml). Based on these results, it is unlikely that these plant extracts would shrink the prostate by inhibiting androgen action or 5 alpha reductase.

Abstract  The hydrogenation of methyl oleate (methyl 9Z-octadecenoate) into oleyl alcohol (9Z-octadecen-1-ol) is carried out in an autoclave at 8.0 MPa and 543 K with Ru-Sn catalysts. The activity and the selectivity of this catalytic system are linked to the preparation method, the nature of the support (alumina, silica, active carbon and zinc oxide) and to the tin content. Selectivities of 50 and 55% to unsaturated alcohol respectively were obtained with a Ru-Sn-B/alumina catalyst resulting from the co-reduction of metallic salts by sodium borohydride and a Ru-Sn/alumina catalyst prepared by a sol-gel method. Our work also showed that a previously unreported transesterification reaction between methyl oleate and oleyl alcohol occurred rapidly and led to a 'higher' ester (oleyl oleate). This side-reaction significantly decreases the selectivity to unsaturated alcohol. Nevertheless, at higher conversion the oleyl oleate is gradually hydrogenated into oleyl alcohol.

Abstract  The conjugation of siRNA to molecules, which can be internalized into the cell via natural transport mechanisms, can result in the enhancement of siRNA cellular uptake. Herein, the carrier-free cellular uptake of nuclease-resistant anti-MDR1 siRNA equipped with lipophilic residues (cholesterol, lithocholic acid, oleyl alcohol and litocholic acid oleylamide) attached to the 5'-end of the sense strand via oligomethylene linker of various length was investigated. A convenient combination of H-phosphonate and phosphoramidite methods was developed for the synthesis of 5'-lipophilic conjugates of siRNAs. It was found that lipophilic siRNA are able to effectively penetrate into HEK293, HepG2 and KB-8-5 cancer cells when used in a micromolar concentration range. The efficiency of the uptake is dependent upon the type of lipophilic moiety, the length of the linker between the moiety and the siRNA and cell type. Among all the conjugates tested, the cholesterol-conjugated siRNAs with linkers containing from 6 to 10 carbon atoms demonstrate the optimal uptake and gene silencing properties: the shortening of the linker reduces the efficiency of the cellular uptake of siRNA conjugates, whereas the lengthening of the linker facilitates the uptake but retards the gene silencing effect and decreases the efficiency of the silencing.

Abstract  Recovery of metabolites from fermentation broth by solvent extraction can be used to optimize fermentation processes. End-product reutilization, low product concentration and large volumes of fermentation broth and the requirements for large bioreactors, in addition to the high cost largely contributed to the decline in fermentative 2,3-butanediol production. Extraction can successfully be used for in-situ alcohol recovery in 2,3-butanediol fermentations to increase the substrate conversion. In the present work organic extraction of 2,3-butanediol produced by Klebsiella pneumoniae fermentation was studied to determine solvent effect on 2,3-butanediol production. The aim of this project was liquid-liquid extractive fermentation systems evaluation as an alternative to overcome the end product effect and to increase of 2,3-butanediol production by K. pneumoniae because Conventional fermentative production of 2,3-butanediol by K. pneumoniae has the disadvantage of product reutilization by the organism. Alternatives to overcome this problem have met with limited success. Extractive fermentation has been shown to solve this problem. An effort has been made in this study to use for the extractive fermentation of 2,3-butanediol using oleyl alcohol as extract-ant. Eighteen organic solvents were examined to determine their biocompatibility for in situ extraction of fermentation products from cultures of the K. pneumoniae. From 18 tested solvents, 13 of which were non-toxic to K. pneumoniae. The highest 2,3-butanediol production (23.01 g l(-1)) was achieved when oleyl alcohol was used. In situ removal of end products from K. pneumoniae resulted in increased productivity. In conclusion 2, 3-hutanediol productivity increased from 0.5 g l(-1)h(-1) to 0.66 g l(-1)h(-1) in extractive fermentation using oleyl alcohol as the extraction solvent.

Abstract  In this work, the potential of waxes for preparing with the ultrasonic spray congealing technique microparticles for controlling the in vitro release of verapamil HCl was investigated. The first part of the study encompassed the optimisation of the formulation to achieve an efficient drug incorporation together with a satisfactory in vitro drug release rate. In particular, microcrystalline wax, stearyl alcohol and mixtures of the two were used. Also a surfactant (soya lecithin) was added to the formulations. After the particle size analysis, the characterisation of the microparticles involved the study of the solid state of drug and carriers in the systems (DSC, HSM and XRD) and the morphological and chemical analyses of the microparticle surface (SEM and XPS). Finally, the drug release mechanism from these devices was evaluated using the statistical moment analysis. The results of this study show that by selecting the type and the amount of the carriers, microparticles with a spherical shape and a good encapsulation efficiency were observed. These particles showed a zero-order release for 8 h, without modifying the solid state properties of the drug. Therefore, waxy microparticles prepared by the ultrasonic spray congealing technique are promising solvent-free devices for controlling the release of verapamil HCl. (C) 2003 Elsevier Science B.V. All rights reserved.

Abstract  To better understand the mode of action of Tadenan, a drug used in the treatment of benign prostatic hyperplasia, the effect of its active principle docosanol, IK.2, was investigated in rats. IK.2 had no effects on the weight and histologic appearance of the prostate in intact rats but increased the RNA/DNA quotient in the ventral prostate. The plasma concentrations of luteinizing hormone and testosterone were reduced. In orchiectomized animals IK.2 increased the weight of the prostate and the adrenals. In adrenalectomized, orchiectomized animals IK.2 did not increase prostatic weight but on the contrary caused a further weight reduction. IK.2 had a thymolytic effect in intact rats but not in adrenalectomized rats in which the thymus weight was increased. The results indicate that IK.2 increases adrenal steroid secretion. The supposedly higher concentration of adrenal androgens causes a stimulation of the prostate most easily discernible in orchiectomized animals. The further weight reduction of the ventral prostate in orchiectomized, adrenalectomized animals, and the increased thymus weight in adrenalectomized animals after IK.2 administration may suggest that IK.2 has effects other than the stimulatory effect on the adrenals.

Abstract  Analysis of the sex pheromone gland of virgin Synanthedon scoliaeformis females by gas chromatography/mass spectrometry revealed six compounds structurally related to sex pheromone components of other clearwing moths: (E,Z)-2,13-octadecadienyl acetate (E2,Z13-18:OAc), (E,Z)-2,13-octadecadienol, octadecanol acetate, octadecanol, (Z,Z)-3,13-octadecadienyl acetate (Z3,Z13-18:OAc), and (Z)-13-octadecenyl acetate. Trapping tests demonstrated that E2,Z13-18:OAc is the sex pheromone of S. scoliaeformis and is essential for attracting males; addition of the other compounds did not enhance catch. Synanthedon scoliaeformis and S. tipuliformis are the only Palearctic clearwing moths whose distribution range and seasonal flight periods overlap and that are known to use E2,Z13-18:OAc in sex pheromonal communication. Hourly monitoring of male catches in traps revealed that sex pheromone communication in S. scoliaeformis and S. tipuliformis species follows different diurnal patterns. Z3,Z13-18:OAc, found in S. scoliaeformis females, is a known behavioral antagonist against S. tipuliformis males, while (E,Z)-3,13-octadecadienyl acetate, a minor sex pheromone component of S. tipuliformis, is an antagonist against S. scoliaeformis males. The effect of sex pheromones and antagonists, combined with different diurnal mate searching times, contribute to the specificity of sex communication channels in these two clearwing moth species.

Abstract  A chemical analysis of the crude sex pheromone gland extracts of virgin calling Paranthrene tabaniformis females, obtained from the European, part of Kazakhstan, revealed the presence of five compounds: (3E,13Z)-octadeca-3,13-dien-1-ol (E3,Z13-18:OH), (3Z,13Z)octadeca-3,13-dien-l-ol (Z3,Z13-18:OH), (2E,13Z)-octadeca-2,13-dien-1-ol (E2,Z13-18:OH), (13Z)-octadec-13-en-1-ol (Z13-18:OH), and octadecan-1-ol (18:OH) at the ratios 64.0:32.4: 1.4:0.9:1.3, which are structurally related to sex pheromone components of clearwing moths. Our previous field tests showed synergistic effects of Z3,Z13-18:OH and E2,Z13-18:OH to attract P tabaniformis males, when these compounds were tested in binary mixtures with the known sex pheromone E3,Z13-18:OH. The three dienic alcohols should all be considered as sex pheromone components of the P tabaniformis species, while the role of Z13-18:OH and 18:011 remained unclear.

Abstract  Interest in pyruvic acid has been growing due to the increase in its potential areas of use and its importance in metabolic reactions. These reasons along with the limitations on recovery have prompted researchers to consider novel recovery techniques. Reactive extraction has been proposed as a promising approach to the recovery of carboxylic acids. In this study, equilibrium and kinetic data were obtained for reactive extraction of pyruvic acid using trioctylamine (TOA) or Alamine 336 in 1-octanol or oleyl alcohol. The results showed that, without pH adjustment in the aqueous phase, and without the use of an extractant, 1-octanol extracted more pyruvic acid than oleyl alcohol with a distribution coefficient (K(D)) of 0.30. This trend remained the same when tertiary amines were used as an extractant. The K(D) values did not significantly differ with TOA or Alamine 336. The recovery of pyruvic acid was observed to increase as a function of TOA concentration and the stoichiometry of the reaction was mainly 1:1. As tertiary amines react only with undissociated acids, an increase in the initial pH of the aqueous phase lowered the KD values. When the pH was 4.0, the effect of TOA concentration on pyruvic acid extraction disappeared and for all concentration levels a distribution coefficient of 0.10 was obtained. Kinetic measurements showed that the reaction between pyruvic acid and TOA in 1-octanol is first order with respect to the two reactants with a rate constant of 0.94 L mol(-1) s(-1). The enhancement factor was calculated as 25.

Abstract  The expression of Pax-5 gene is altered in human myeloma cells (malignant plasma cells). This altered expression is considered to be closely involved in oncogenesis of human myeloma. To investigate the possible mechanism(s) underlying this alteration, we first cloned the 1,050 bp fragment in the 5' upstream region of human Pax-5 gene by PCR-mediated gene walking method. The cloned fragment has predicted regulatory motifs for Lyf-1(Ik-1), IK-2, bHLH, E-47, Sox-5, Oct-1, GATA-1,-2, and -3, but it lacks a TATA box. By constructing deletion mutants of this fragment, its basal promoter activity was analyzed by transfecting these mutants to Cos 7 cells. The maximal promoter activity was recovered by the fragment that extends between -70 to -820 upstream of the transcription start site. Also, three DNA fragments from this cloned sequence were used as templates in gel shift assay; these fragments covered most of the predicted regulatory sites. Specific binding activities were found in each DNA fragment. Therefore, we could clone the functionally active fragment of 5' upstream region of human Pax-5 gene.

Abstract  Herpes simplex virus type 1 (HSV-1) is a causative agent for a variety of diseases. Although antiherpetic drugs such as acyclovir have been developed to inhibit virus replication through interaction with DNA kinases, their continuous administration leads to an increase in the frequency of drug-resistant HSV-1, which is an important clinical issue that requires urgent solution. Recently, we reported that the sialylated O-linked sugar T antigen (sTn) and its attached peptide region (O-glycosylated sTn peptide) derived from the HSV-1 glycoprotein B (gB) protein inhibited HSV-1 infection by specifically targeting paired immunoglobulin-like type 2 receptor alpha (PILRα) in vitro. In this study, to further identify novel inhibitors of gB-mediated HSV-1 infection in vitro, we established a cell-based fusion assay for rapid drug screening. Chinese hamster ovary (CHO) cells were transfected with expression plasmids for HSV-1 gB, gD, gH, and gL, and T7 RNA polymerase, and were designated as the effector cells. The CHO-K1 cells stably expressing PILRα were transfected with the expression plasmid for firefly luciferase under the T7 promoter, and were designated as the target cells. The effector and target cells were co-cultured, and luminescence was measured when both cells were successfully fused. Importantly, we found that cell-to-cell fusion was specifically inhibited by O-glycosylated sTn peptide in a dose dependent manner. Our results suggested that this virus-free cell-based fusion assay system could be a useful and promising approach to identify novel inhibitors of gB-mediated HSV-1 infection, and will aid in the development of antiviral therapeutic strategies for HSV-1-associated diseases.

Abstract  Ether lipids show high specific cytotoxicity in vitro on a wide variety of experimental tumors, but only moderate activity in vivo. One reason for this lack of activity in the whole animal might be a high degree of metabolic degradation. We therefore studied the biotransformation of 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine ([H]ET18-OMe) labeled in position 9-10 of the 1-alkyl chain, in rat plasma and erythrocytes, HL60 and K562 leukemic cells. HT29 adenocarcinoma cells, and cultured hepatocytes at 37 degrees C, and in a system of isolated and perfused rat liver. ET18-OMe and its metabolites were identified and quantified after lipid extraction and TLC separation. In tumor cells, 98% of ET18-OMe remained almost unmodified in vitro after 24-hr incubation. Plasma and erythrocytes from rats metabolized only 4-5% of the original compound in 3 hr. In cultured hepatocytes, 35% and 58.3%, respectively, of ET18-OMe was present after 6 and 24 hr as the metabolites 1-O-alkyl-2-O-methylglycerol (AMG), 1-O-alkyl-2-O-methylphosphatidic acid (AMPA), and stearyl alcohol (SA) (products of direct hydrolysis by phospholipases C and D and alkylhydrolase); phospholipids (phosphatidylcholine and phosphatidylethanolamine); and neutral lipids (products of secondary metabolism). In perfused rat liver, similar to 15% of the total radioactivity incorporated after 3 hr was distributed in metabolites as follows: 5.9% of AMPA, 5.0% of AMG, and 3.1% of SA. We conclude that the metabolism of ET18-OMe in normal tissues occurring through the same enzymes that metabolize natural lipids may partly explain the lack of effect in vivo.

Abstract  Objective To systematically review the evidence for the clinical effects and safety of the rye-grass pollen extract (Cernilton) in men with symptomatic benign prostatic hyperplasia (BPH).

Methods Trials were identified by searching Medline, specialized databases (EMBASE, Cochrane Library, Phytodok), bibliographies, and contacting relevant trialists and manufacturers. Randomized or controlled clinical trials were included if: men with symptomatic BPH were treated with Cernilton; a control group received either placebo or pharmacological therapy; the treatment duration was greater than or equal to 30 days; and clinical outcomes were reported.

Results In all, 444 men were enrolled in two placebo-controlled and two comparative trials lasting 12-24 weeks. Three studies used a double-blind method although the concealment of treatment allocation was unclear in all. Cernilton improved 'self-rated urinary symptoms' (the proportion reporting satisfactory or improving symptoms) vs placebo and another plant product, Tadenan. The weighted mean (95% confidence interval) risk ratio (RR) for self-rated improvement vs placebo was 2.40 (1.21-4.75) and the weighted RR vs Tadenan was 1.42 (1.21-4.75). Cernilton reduced nocturia compared with placebo or Paraprost (a mixture of amino acids); against placebo, the weighted RR was 2.05 (1.41-3.00), and against Paraprost the weighted mean difference for nocturia was - 0.40 times per evening (-0.73 to 0.07). Cernilton did not improve urinary flow rates, residual volume or prostate size compared with placebo or the comparative study agents. Adverse events were rare and mild; the withdrawal rate for Cernilton was 4.8%, compared with 2.7% for placebo and 5.2% for Paraprost.

Conclusions The Cernilton trials analysed were limited by their short duration, limited number of enrolees, omissions in reported outcomes, and the unknown quality of the preparations used. The comparative trials had no confirmed active control. The available evidence suggests that Cernilton is well tolerated and modestly improves overall urological symptoms, including nocturia. Additional randomized placebo and active-controlled trials are needed to evaluate the long-term clinical effectiveness and safety of Cernilton.

Abstract  A goldfish (Carassius auratus) model of Parkinson's disease (PD) was constructed by a single dose of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) according to previously reported methods. Global metabolite changes in brain of the MPTP induced goldfish model of PD were investigated. (1)H NMR-based metabolomics combined with various statistical methods such as orthogonal partial least squares discriminant analysis (OPLS-DA) and two-dimensional statistical total correlation spectroscopy (2D-STOCSY) found significant increase of leucine, isoleucine, valine, alanine, alanylalanine, creatinine, myo-inositol, 18:2 fatty acid, total fatty acids, arachic alcohol, taurine and significant decrease of N-acetylaspartate, (phospho)creatine, (phospho)choline, betaine, glutamine, 3-hexenedioate, acetamide, malonate, isocitrate, scyllo-inositol, phosphatidylcholines, cholesterols, n-3 fatty acids, polyunsaturated fatty acids (PUFAs) in brain of MPTP induced PD goldfish. These disturbed metabolite levels were involved in oxidative stress, energy failure, neuronal cell injury and death, consistent with those observed in clinical PD patients, and rodents and primates model of PD, indicating that the acute MPTP model of goldfish was an ideal and valuable model for PD research. In addition, several unusual metabolites in brain were significantly changed between MPTP induced PD and control goldfish, which might also play an important role in the pathogenesis of PD. This study also demonstrated the applicability and potential of (1)H NMR-based metabolomics approach for evaluation of animal models of disease induced by chemicals, such as MPTP-induced PD goldfish.

Abstract  Purpose: Tadenan is a pharmaceutical agent used in the treatment of BPH. Prior studies demonstrated that pretreatment of rabbits with Tadenan significantly reduced the contractile dysfunction following two weeks of partial outlet obstruction, The specific aim of the present study was to determine the effect of Tadenan pretreatment on the time course of the response to partial outlet obstruction and correlate the effect of Tadenan on the contractile responses to field stimulation, bethanechol, and KCl with both mitochondrial enzyme activity (citrate synthase) and sarcoplasmic reticular function (calcium-ATP'ase activity).

Materials and Methods: Sixty male New Zealands white rabbit (3 to 5 kg.) were separated into 12 groups of 5 rabbits each. Each rabbit in groups 1-6 received Tadenan orally at 100 mg./kg./day for three weeks; each rabbit in groups 7-12 received vehicle (peanut oil). Each rabbit in groups 2-6 and 8-12 received a partial outlet obstruction as described below. One group of Tadenan treated and one group of vehicle-treated rabbits were euthanized at 1, 3, 5, 7, and 14 days following partial outlet obstruction. The non-obstructed groups were studied after 4 weeks of drug or vehicle treatment. Each bladder was rapidly removed and weighed, and 3 longitudinal strips prepared and mounted in individual baths for contractile studies. The remainder of the bladder was frozen for biochemical analysis. The contractile responses to field stimulation, bethanechol, and KCl were determined; and the enzyme activities of citrate synthase (marker for mitochondrial function) and calcium-ATP'ase (marker for sarcoplasmic reticulum) were determined,

Results: 1) Tadenan did not reduce the effect of partial outlet obstruction on bladder mass. 2) Although the contractile responses to all forms of stimulation were reduced at 1 day following partial outlet obstruction, Tadenan pretreatment resulted in a significant protective effect on the contractile responses to field stimulation, bethanechol, and KCl at 3, 5, 7, and 14 days of obstruction. 3) The activities of both citrate synthase and calcium ATP'ase were reduced significantly at 1 day following obstruction for both Tadenan treated and vehicle treated groups. The activities of both enzymes returned to near normal levels at 7 and 14 days for the Tadenan groups whereas the activities of both enzymes remained significantly reduced in the vehicle treated groups.

Conclusions: These results clearly demonstrate that Tadenan pretreatment protected the bladder from both the contractile and metabolic dysfunctions induced by partial outlet obstruction.