Auditory-verbal graduates: outcome survey of clinical efficacy.

This project is an update of an earlier study on American and Canadian graduates of auditory-verbal programs. Survey research was conducted to obtain information on a variety of topics. Overall, the current results again indicated that the majority of respondents were integrated into "regular" or "typical" learning and living environments. In view of the earlier identification of hearing loss and the early fitting of sensory aids and availability of cochlear implant technology, coupled with intervention that emphasizes auditory learning, it is suggested that today's infants have the potential to become independent, participating, and contributing citizens in mainstream society.

Do wine polyphenols modulate p53 gene expression in human cancer cell lines?

BACKGROUND: The p53 gene is an established tumor suppressor and an inducer of apoptosis. We here attempt to determine whether the putative anticarcinogenic properties attributed to red wine and its polyphenolic constituents depend, at least in part, upon their ability to modulate p53 expression in cancer cells. METHODS: Three human breast cancer cell lines (MCF-7, T47D; MDA-MB-486) and one human colon cancer cell line [Colo 320 HSR (+)] were treated for 24-h with each of four polyphenols [quercetin; (+)-catechin, trans-resveratrol; caffeic acid] at concentrations ranging from 10(-7) M to 10(-4) M, after which, p53 concentrations were measured in cell lysates by a time-resolved fluorescence immunoassay. RESULTS: None of the polyphenols tested affected p53 expression in the breast cancer cell lines T-47D and MDA-MB-486. p53 content of MCF-7 breast cancer cells (wild-type) was increased by caffeic acid, decreased by resveratrol, and showed a twofold increase with catechin, that reached borderline statistical significance; however, none of these effects were dose-responsive. Colo 320 HSR (+) cells (with a mutant p53 gene) had lower p53 content upon stimulation, reaching borderline statistical significance, but without being dose-responsive, in the presence of caffeic acid and resveratrol. Apart from toxicity at 10(-4) M, quercetin had no effect upon these four cell lines. CONCLUSIONS: The observed p53 concentration changes upon stimulation by polyphenols are relatively small, do not follow a uniform pattern in the four cell lines tested, and do not exhibit a dose-response effect. For these reasons, we speculate that the putative anticarcinogenic properties of wine polyphenols are unlikely to be mediated by modulation of p53 gene expression.

Assay of ochratoxin A in wine and beer by high-pressure liquid chromatography photodiode array and gas chromatography mass selective detection.

To routinely assay the concentrations of ochratoxin A (OTA) in wines and beers, two new methods were developed and evaluated. The first utilized solid-phase extraction on a C(18) cartridge to achieve a 100-fold sample concentration followed by high-performance liquid chromatography on a C(18) column with gradient elution and quantitation at 333 nm by means of a photodiode array detector. Positive confirmation can be carried out by purity and match-factor analysis as well as peak shift following esterification with BF(3). Total run time is 28 min. The limits of detection (LOD) and quantitation (LOQ) are 0.05 and 0.10 microg/L, respectively. Recovery and imprecision ranged from 83 to 94% and from 4.0 to 8.9%, respectively. With a throughput of 35 assays per working day, this method is ideal for routine OTA analysis. It was used to survey the concentrations of OTA in 942 wines (2 of which gave values between 0.1 and 0.2 microg/L) and 107 beers (2 of which gave values between 0.05 and 0.1 microg/L). OTA was detected more frequently in red than white wines, with the highest incidence in red wines from Spain and Argentina. There was no association between OTA and country of origin or beverage type among the beers analyzed. The second method utilized gas chromatography with mass selective detection monitoring eight specific ions, preceded by extraction in dichloromethane and derivatization with bis[trimethylsilyl]trifluoroacetamide. LOD and LOQ were 0.1 and 2 microg/L, respectively; recovery and imprecision were 69-75 and 9.0-11.1%, respectively. The method is not suitable for routine quantitation but is potentially useful as a confirmatory tool for samples with OTA > or =0.1 microg/L.

Ultrasensitive assay for three polyphenols (catechin, quercetin and resveratrol) and their conjugates in biological fluids utilizing gas chromatography with mass selective detection.

The concentrations of three polyphenols ((+)-catechin, quercetin and trans-resveratrol) in blood serum, plasma and urine, as well as whole blood, have been measured after their oral and intragastric administration, respectively, to humans and rats. The method developed for this purpose utilized ethyl acetate extraction of 100 microl samples and their derivatization with bis(trimethylsilyl)trifluoroacetamide (BSTFA) followed by gas-chromatographic analysis on a DB-5 column followed by mass selective detection employing two target ions and one qualifier ion for each compound. Total run time was 17 min with excellent resolution and linearity. The limits of detection (LOD) and quantitation (LOQ) were an order of magnitude less than for any previously published method, being 0.01 microg/l and 0.1 microg/l, respectively, for all compounds. Recovery at 1 microg/l and 10 microg/l was >80% in all instances but one, and was >90% in 50%. Imprecision was acceptable at 0.25 and 1.0 microg/l, concentrations below the LOQ of previous methods. Aglycones released from conjugates after hydrolysis were easily measurable. Optimal conditions for hydrolysis were established. After oral administration of the three polyphenols to humans, their conjugates vastly exceeded the concentrations of the aglycones in both plasma and urine. Concentrations peaked within 0.5-1.0 h in plasma and within 8 h in urine. During the first 24 h, 5.1% of the (+)-catechin and 24.6% of the trans-resveratrol given were recovered in the urine (free plus conjugated). This method can be proposed as the method of choice to assay these polyphenols and their conjugates in biological fluids.

Multiresidue analysis of seventeen pesticides in wine by gas chromatography with mass-selective detection.

We have developed a multiresidue method permitting the simultaneous quantitation of 17 pesticides in wine: dicloran, dimethoate, diazinon, chlorpyrifos-methyl, vinclozolin, carbaryl, methiocarb, dichlofluanid, parathion-ethyl, triadimefon, procymidone, myclobutanil, iprodione, imidan, dicofol, phosalone and azinphos-methyl. Solid-phase extraction of 0.5 ml of wine sample is followed by direct injection of 1 microl of the eluent onto a DB-5 MS gas chromatographic column followed by mass-selective detection using one target and two qualifier ions for each pesticide. The extraction and injection steps are carried out with automatic instrumentation. Good resolution of all compounds was achieved with a run-time approximating 23 min. Detection and quantitation limits were around 2 microg/l and 10 microg/l, respectively, with linear calibration curves up to 3 mg/l for most constituents. Recovery in half the compounds was >90%, and >80% in most of the remainder. Imprecision (relative standard deviation) was <10% for most pesticides and <18% in all. Further analytes can be added to the repertoire without difficulty. The method merits consideration together with four other multiresidue methods now available that offer similar analytical characteristics, slower run-times, and a different selection of analytes.

Proteases in the evaluation of pancreatic function and pancreatic disease.

This paper reviews the role of pancreatic proteases (focusing upon trypsin, chymotrypsin and elastase) in the diagnosis and management of chronic pancreatic insufficiency (CPI), emphasizing advances over the last 5 years. Some important novel aspects of these enzymes in acute pancreatitis are also described, including their role in diagnosis and their interaction with cholecystokinin in the pathogenesis of the disease. The recent interest in these enzymes as agents promoting the spread of cancer in animals and human subjects is also described. A hierarchical approach has been taken to explore the advantages and limitations of tests in different source materials: serum, feces, duodenal aspirate, and non-invasive pancreatic function tests. The practical usefulness of fecal elastase-1 and of fecal chymotrypsin concentrations in diagnosis and management of CPI, respectively, is one of the major lessons to be learned from analysis of the recent literature, and forms the principal message of this review.

Phenolic constituents, furans, and total antioxidant status of distilled spirits.

The concentrations of 11 phenols and 5 furans were measured in 12 categories of distilled spirits by HPLC methodology, together with the total antioxidant status (TAS) of the same beverages. Ellagic acid was the phenol present in highest concentration in all beverages. Moderate amounts of syringaldehyde, syringic acid, and gallic acid, as well as lesser amounts of vanillin and vanillic acid, were measurable in most samples of whiskey, brandy, and rum but were largely undetectable in gin, vodka, liqueurs, and miscellaneous spirits. 5-(Hydroxymethyl)furfural was the predominant furan in the former three beverages, notably cognac, with 2-furaldehyde the next highest, but these were undetectable in most of the latter beverages. Highest TAS values were given by armagnac, cognac, and bourbon whiskey, all three of which tended toward the highest concentrations of phenols. Negative TAS values were exhibited by rum, vodka, gin, and miscellaneous spirits in line with the low or undetectable phenol concentrations in these beverages. Wood aging is the most likely source of phenols and furans in distilled spirits. Those beverages exposed to this treatment contain significant antioxidant activity, which is between the ranges for white and red wines, with the potential to augment the antiatherosclerotic functions attributable to the ethanol that they contain.

Moderate alcohol consumption: the gentle face of Janus.

OBJECTIVES: The regular consumption of alcohol in moderate amounts (defined in North America as up to 2 drinks per day for men and 1 drink per day for females) has been recognized in the last decade as a negative risk factor for atherosclerosis and its clinical sequelae: coronary heart disease (CHD), ischemic stroke, and peripheral vascular disease. Mortality and morbidity attributable to CHD are 40-60% lower in moderate drinkers than among abstainers. Among the mechanisms accounting for these reductions, increased circulating concentrations of HDL-cholesterol and inhibition of blood coagulation appear to be paramount. Additional benefits are, in certain beverages, conferred by the presence of constituents other than alcohol (e.g., flavonoids and hydroxystilbenes), which prevent oxidative damage, free radical formation, and elements of the inflammatory response. CONCLUSIONS: A number of other diseases appear to be beneficially modulated by moderate alcohol consumption based on epidemiologic surveys and, in some instances, experimental evidence. These include duodenal ulcer, gallstones, enteric infections, rheumatoid arthritis, osteoporosis, and diabetes mellitus (type II). Compared with abstainers, moderate drinkers exhibit improved mental status characterized by decreased stress and depression, lower absenteeism from work, and decreased incidence of dementia (including Alzheimer's disease). Although limits of safe drinking have been conservatively defined, it is regrettable that political considerations are hampering the clinical application of this knowledge and its dissemination to the lay public.

Changes in serum carbohydrate-deficient transferrin and gammaglutamyl transferase after moderate wine consumption in healthy males.

Serum carbohydrate-deficient transferrin (CDT) concentrations and gammaglutamyl transferase (GGT) activities were measured in the fasting serum of healthy male subjects before and after 4 weeks consumption each day of 375 ml wine or 500 ml grape juice. After wine consumption, serum CDT concentrations rose in 38 of 48 individual test procedures, and the mean +/- SEM increased from 17.8 +/- 0.86 u/l to 20.9 +/- 1.14 u/l (t0 = 4.66; P < 0.001). Serum GGT activity rose in 35 of these test procedures, and the mean +/- SEM increased from 19.6 +/- 1.40 u/l to 22.3 +/- 1.79 u/l (t0 = 3.58; P < 0.001). When wine consumption was followed by 2 weeks of abstinence from alcohol, significant reductions in both CDT and GGT were noted, virtually reaching baseline levels. No significant change in either index occurred after 4 weeks of consuming grape juice. The correlation between CDT and GGT was rather low, suggesting that their responses to alcohol occur by different mechanisms. The results indicate that the response of CDT to alcohol dose is continuous, and that even moderate consumption can cause significant elevations in a healthy population.

Biochemical tests in diseases of the intestinal tract: their contributions to diagnosis, management, and understanding the pathophysiology of specific disease states.

Biochemical testing plays a major role in the complete evaluation of patients with suspected or established intestinal disease. We have classified these tests according to the medium in which they are performed: breath tests, including isotopic and nonisotopic tests, fecal tests, urine tests, serum tests, tissue tests, and other tests. The principles of various tests are outlined, and the role of each test in the evaluation of particular gastrointestinal disorders is discussed.

Resveratrol: a molecule whose time has come? And gone?

OBJECTIVES: Resveratrol (3,5,4'-trihydroxystilbene) is the parent compound of a family of molecules, including glucosides and polymers, existing in cis and trans configurations in a narrow range of spermatophytes of which vines, peanuts and pines are the prime representatives. Its synthesis from p-coumaroyl CoA and malonyl CoA is induced by stress, injury, infection or UV-irradiation, and it is classified as a phytoalexin anti-fungicide conferring disease resistance in the plant kingdom. RESULTS: In vitro, ex vivo and animal experiments have shown that it possesses many biological attributes that favour protection against atherosclerosis, including antioxidant activity, modulation of hepatic apolipoprotein and lipid synthesis, inhibition of platelet aggregation as well as the production of pro-atherogenic eicosanoids by human platelets and neutrophils. Red wine represents its main source in the human diet, and it has been proposed as a major constituent of the polyphenol fraction to which the health benefits of red wine consumption have been attributed. CONCLUSIONS: The past several years have witnessed intense research devoted to its measurement in wine and the factors likely to promote its enrichment in this beverage. Up to the present, conclusive evidence for its absorption by human subjectsin biologically significant amounts is lacking, and it is questionable (but not yetexcluded) that its powerful and beneficial in vitro activities are reproduced as a consequence of sustained moderate red wine consumption.

Wine as a biological fluid: history, production, and role in disease prevention.

Wine has been part of human culture for 6,000 years, serving dietary and socio-religious functions. Its production takes place on every continent, and its chemical composition is profoundly influenced by enological techniques, the grape cultivar from which it originates, and climatic factors. In addition to ethanol, which in moderate consumption can reduce mortality from coronary heart disease by increasing high-density lipoprotein cholesterol and inhibiting platelet aggregation, wine (especially red wine) contains a range of polyphenols that have desirable biological properties. These include the phenolic acids (p-coumaric, cinnamic, caffeic, gentisic, ferulic, and vanillic acids), trihydroxy stilbenes (resveratrol and polydatin), and flavonoids (catechin, epicatechin, and quercetin). They are synthesized by a common pathway from phenylalanine involving polyketide condensation reactions. Metabolic regulation is provided by competition between resveratrol synthase and chalcone synthase for a common precursor pool of acyl-CoA derivatives. Polymeric aggregation gives rise, in turn to the viniferins (potent antifungal agents) and procyanidins (strong antioxidants that also inhibit platelet aggregation). The antioxidant effects of red wine and of its major polyphenols have been demonstrated in many experimental systems spanning the range from in vitro studies (human low-density lipoprotein, liposomes, macrophages, cultured cells) to investigations in healthy human subjects. Several of these compounds (notably catechin, quercetin, and resveratrol) promote nitric oxide production by vascular endothelium; inhibit the synthesis of thromboxane in platelets and leukotriene in neutrophils, modulate the synthesis and secretion of lipoproteins in whole animals and human cell lines, and arrest tumour growth as well as inhibit carcinogenesis in different experimental models. Target mechanisms to account for these effects include inhibition of phospholipase A2 and cyclo-oxygenase, inhibition of phosphodiesterase with increase in cyclic nucleotide concentrations, and inhibition of several protein kinases involved in cell signalling. Although their bioavailability remains to be fully established, red wine provides a more favourable milieu than fruits and vegetables, their other dietary source in humans.

Genetic markers of alcohol abuse.

In this paper, we review the current status of genetic markers for the development of alcohol abuse. Family, twin, half-sibling and adoption studies of alcoholic subjects suggest that the heritability of liability to alcoholism is at least 50%. These findings have fuelled intensive investigation in the fields of neurology, biochemistry, genetics and molecular biology aimed at the identification of markers for the risk of alcoholism. The most promising of these are discussed in detail. Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) polymorphisms, specifically the ADH3*1, ADH2*2, and ALDH2*2 genotypes appear to confer a protective effect against alcoholism, most notably in Oriental subjects. Caucasian alcohol abusers and their first-degree relatives exhibit depressed platelet monoamine oxidase activity, the degree of which is greater in Type II than Type I alcoholics. Electrophysiological characteristics of alcoholics and those at risk for developing alcoholism have also been identified, including the reduced amplitude of the event-related brain potential and, after ethanol ingestion, characteristic EEG alpha-wave activity. Lower platelet adenylate cyclase activity is seen in alcoholics compared to controls, presumably as a result of over-expression of an inhibitory G-protein. Markers related to other signal transduction pathways of the central nervous system including the serotoninergic, muscarinic and dopaminergic systems are also discussed. In this group of markers, the putative association between the inheritance of the AI allele of the D2 dopamine receptor and the susceptibility to alcoholism provides the most dramatic illustration of the challenges presently existing in this field of scientific investigation. Current limitations in the definition, diagnosis and classification of alcoholism, the confounding influences of race and gender on association studies, as well as the statistical approach of linkage studies are discussed as they relate to the endeavor to uncover valid genetic markers for the risk of alcoholism.

A multiresidue derivatization gas chromatographic assay for fifteen phenolic constituents with mass selective detection.

We have developed a GC/MS method to simultaneously measure the concentrations of 15 biologically active phenolic components of wine: vanillic acid, gentisic acid, m- and p-coumaric acid, gallic acid, ferulic acid, caffeic acid, cis- and trans-resveratrol, epicatechin, catechin, morin, quercetin, and cis- and trans-polydatin. Wine (1 mL) was diluted 1:1 with water to reduce the alcohol content and extracted on a preconditioned C-8 solid-phase extraction cartridge. The phenolic compounds were eluted with ethyl acetate, evaporated to dryness, and derivatized with bis(trimethylsilyl)trifluoroacetamide/pyridine. The TMS derivative of each phenolic compound was analyzed on a GC/MSD coupled to a DB-5HT capillary column using one target and two qualifying ions for each compound in a total run time of 26 min. Resolution and quantitation of all compounds were excellent, with linear calibration curves over a wide range. The lowest detection limit was for gentisic acid (24 µg/L) and highest for quercetin (843 µg/L). The average percent recovery and coefficient of variation (mean precision) ranged from 90.7 to 104.6 (except morin, 72.2%) and 4.0 to 10.2 (except morin, 16.1%, and quercetin, 16.0%) respectively. This method has been applied to solid vitaceous plant materials as well as wine and should be suitable to measure polyphenols in fruit, vegetables, and other foods provided that efficient extraction techniques are employed.

Method to assay the concentrations of phenolic constituents of biological interest in wines.

We describe a reversed-phase HPLC method that uses gradient elution and diode array detection to quantitate eight biologically active phenolic constitutions of wine: the cis and trans isomers of resveratrol and their glucosides, catechin, epicatechin, quercetin, and rutin. ODS Hypersil served as the stationary phase; the gradient was formed by acetic acid, methanol and water. Each analysis required an equilibration period of ten minutes and a run time of fourty minutes for completion. Satisfactory peak resolution was achieved following direct injection of a 20-muL sample, and validation was accomplished by on-line spectral comparisons with known standards. Excellent linearity was obtained for all constituents, and the detection limits ranged from 30 mug/L (trans-resveratrol) to 1.5 mg/L (catechin). Recoveries approximated 100% range (95.2-105.5%), and the method provided good precision, with coefficients of variation between 1.17 and 3.38%. All of the phenolics measured were reasonably stable in opened wines protected against sunlight for up to 1 week at room temperature or 4 degrees C, but most showed losses of 10-40% when stored for 6 weeks at either temperature.