Christopher W. Shade, PhD: Defining Cannabinoids.

No Abstract Available.

Liposomes as Advanced Delivery Systems for Nutraceuticals.

Liposomes are delivery vehicles for transporting substances into the body effectively via facilitating absorption directly in the mouth or by preventing breakdown by stomach acid. Since the 1970s, liposomes have been investigated as potential drug delivery systems because of their biocompatibility and ability to incorporate both hydrophilic and hydrophobic therapeutic agents. Despite early promise, it was decades later, in the late 1990s to the present, that liposome technologies could create successful commercial products. Oral deliveries are recently emerging as availability of quality phospholipids and reliable homogenization and sizing equipment have become routinely available. Nutritional industry use of liposomes will grow rapidly in the next 5-10 y. High-quality products with more complex mixtures of pure compounds and complex botanical mixtures will offer clinicians less-invasive options for dosing and delivery of these actives.

A double-blind, randomized study to assess the validity of applied kinesiology (AK) as a diagnostic tool and as a nonlocal proximity effect.

PREMISE: Applied Kinesiology (AK) is a diagnostic technique widely used within the Integrative Medical community. In essence, it posits that a question can be mentally held in a person's mind, sometimes while they are holding a substance like a vitamin, or a food sample, and by measuring relative muscular weakness an answer as to whether the substance or the condition represented by the question is good for that person can be obtained. This AK is presumed to have a diagnostic capability. That being presumed, this study asks the following questions: (1) Is there a difference in muscular strength when an individual holds a substance that is inimical to life processes (a poison solution), as compared to a substance that is essential for life (normal saline)? (2) Is this effect a transaction involving input from both the person being measured and the kinesiologist doing the measurement or is it only the person being measured? (3) As an extension of question 2, is the result the same when different kinesiologists take the measurement or when no kinesiologist is involved? (4) Does belief, expectation, gender, or time cognition play a role in determining response? METHODOLOGY: To answer these questions, which would help to define the parameters of the AK process, 51 participants were tested during three trials each, first by one kinesiologist, then by another, and finally, with no kinesiologist present by grip strength indicated using a hand dynamometer. Grip strength being a self-administered AK test of relative muscular strength. For each trial, a pair of randomly numbered sealed vials, each pair in a randomly numbered plastic bag, were used as the objects of the trial. In each bag, one vial contained saline solution while the other was filled with a slightly smaller amount of saline solution to which had been added ionic hydroxylamine hydrochloride (NH3OH)(+), producing a toxic solution of 9mg/ml. Each trial consisted of a separate muscle test for each vial. All present at the trials were blind as to which vial contained the toxin. And all who prepared the vials were blind to the trials. The force used by the kinesiologists in each of their trials was measured via a pressure pad system. The hand dynamometer trials were conducted with no kinesiologist present. RESULTS: Of the 151 sets of trials, the toxic vial was identified correctly in 80 of them (53%), resulting in a one-tailed exact binomial P-value of .258. Results for two of the kinesiologists were almost exactly at chance. For the third kinesiologist, there was a one-tailed exact binomial P-value of .18 (unadjusted for multiple testing). Results for the dynamometer were also almost exactly at chance. Testing whether there was a significant difference in proportions for whom the AK test worked based on belief about whether it would work resulted in non-significant χ(2) values of 0.6 (P = .439) for the trials with one kinesiologist and 2.222 (P = .136) for the hand dynamometer trials. The final variable examined was gender. While there was no significant difference in performance for males and females for the trials of the male kinesiologist or the hand dynamometer, the combined data for the two female kinesiologists did reveal a difference. Of the 33 sessions with females, only 15 were successful (45%), while for the 18 sessions with males, 14 were successful (78%), resulting in a χ(2) statistic of 4.96, P = .026. However, given all of the χ(2) tests performed in this section, the results must be interpreted with caution because of multiple testing. Results indicate belief in whether the AK test will work was not significantly related to whether it actually did work. A χ(2) test of the relationship between time perception and correct vial choice showed no significant relationships. The χ(2) statistic for the relationship using the hand dynamometer data was 0.927, P = .629. CONCLUSION: The data in this study, particularly when seen in the larger context of a review of the literature from the AK field itself by Klinkoski and Leboeuf (1990), which considered 50 papers published between 1981 and 1987 by the International College of Applied Kinesiology, and the survey by Hall, Lewith, Brien, and Little (2008), using standard evaluation criteria [quality assessment tool for studies of diagnostic accuracy included in systematic reviews (QUADAS), Standards for Reporting of Diagnostic Studies (STARD), JADAD, and Consolidated Standards of Reporting Trials (CONSORT)], for research methodology, as well as six prior non-clinical studies by Radin (1984), Quintanar and Hill (1988), Braud (1989), Arnett et al. (1999), Ludtke (2001), and Kendler and Keating (2003), all together suggest the following: The research published by the Applied Kinesiology field itself is not to be relied upon, and in the experimental studies that do meet accepted standards of science, Applied Kinesiology has not demonstrated that it is a useful or reliable diagnostic tool upon which health decisions can be based.

Automated speciation of mercury in the hair of breastfed infants exposed to ethylmercury from thimerosal-containing vaccines.

A simplified thiourea-based chromatography method, originally developed for methyl and inorganic mercury, was adapted to separate methylmercury (MeHg), ethylmercury (EtHg), and inorganic mercury (Hg(II)) in infants' hair. Samples were weighed and leached with an acidic thiourea solution. Leachates were concentrated on a polymeric resin prior to analysis by Hg-thiourea liquid chromatography/cold vapor atomic fluorescence spectrometry. All but one sample showed small amounts of EtHg, and four of the six analyzed samples had proportionally higher Hg(II) as a percent of total Hg. Breastfed infants from riverine Amazonian communities are exposed to mercury in breast milk (from high levels of maternal sources that include both fish consumption and dental amalgam) and to EtHg in vaccines (from thimerosal). The method proved sensitive enough to detect and quantify acute EtHg exposure after shots of thimerosal-containing vaccines. Based on work with MeHg and Hg(II), estimated detection limits for this method are 0.050, 0.10, and 0.10 ng g⁻¹ for MeHg, Hg(II), and EtHg, respectively, for a 20-mg sample. Specific limits depend on the amount of sample extracted and the amount of extract injected.

Bioaccessibility of mercury from traditional northern country foods measured using an in vitro gastrointestinal model is independent of mercury concentration.

Human health risk assessment of dietary mercury (Hg) exposure in Canada assumes that all Hg from fish consumption is in the form of methylmercury (MeHg), the more bioavailable and hazardous form of Hg. In contrast, the risk assessment of dietary Hg to Inuit in northern Canada assumes that no more than two-thirds of dietary Hg is MeHg since mammal organs consumed by Inuit contain substantial concentrations of inorganic Hg. In vitro gastrointestinal models (e.g., the Simulator of the Human Intestinal Microbial Ecosystem) are often used for the evaluation of soil contaminant bioaccessibility, i.e., the fraction solubilized into gastrointestinal fluids, for use in site-specific human health risk assessment. In this research, we digested northern country foods using the SHIME for the measurement of Hg bioaccessibility, a novel approach for the assessment of dietary Hg bioavailability. We demonstrated that small intestinal Hg bioaccessibility from 16 fish, wild game, and marine mammal samples consumed by Inuit in northern Canada ranged between 1 and 93% and was independent of food HgT (MeHg+Hg(II)) concentration. Additionally, we demonstrated that gastrointestinal microbes may affect Hg bioaccessibility of the 16 country foods, either increasing or decreasing bioaccessibility depending upon the type of food. These results indicate that gastrointestinal absorption of Hg is not likely limited by the concentration of Hg in the food, which is in agreement with in vivo Hg bioavailability studies. Furthermore, these in vitro results support the hypothesis that the gastrointestinal absorption of Hg from Inuit country foods is dependent upon food type.

Automated simultaneous analysis of monomethyl and mercuric Hg in biotic samples by Hg-thiourea complex liquid chromatography following acidic thiourea leaching.

A simple leaching procedure has been validated for quantitative isolation of both monomethyl (CH3Hg+) and inorganic (Hg(II)) mercuryfrom fresh or dried biotic tissue for simultaneous analysis via separation and quantification with Hg-thiourea complex liquid chromatography cold vapor atomic fluorescence spectrometry (HgTU/LC-CVAFS). The leaching solution comprises thiourea, hydrochloric acid, and glacial acetic acid and works by protonating thiol binding sites and forming water-soluble cationic CH3HgS=C(NH2)2+ and Hg[S=C(NH2)2]2(2+) complexes, which are easily separated from the solid matrix. The isolated complexes are preconcentrated online by either thiol resin trapping or a new iodide-complex polydivinylbenzene resin trapping (I-PDVB). The I-PDVB trapping involves only one reagent addition, requires no pH adjustments, and is quantitative over a large range of volumes and flow rates. The chromatography system can use either ion chromatography or a new ion-pairing reversed phase separation coupled to cold vapor generation and atomic fluorescence detection. The system allows quantitative sample introduction and yields absolute detection limits of 0.4 pg and 0.7 pg, for CH3Hg+ and Hg(II) respectively, enabling relative detection limits as low as 4 and 7 pg g(-1) with 100 mg samples, and yields % CV routinely less than 5% with well homogenized samples. Accuracy for both forms of mercury has been validated with multiple biotic reference materials and by comparison of the sum of CH3Hg+ and Hg(II) with total Hg on a variety of different biotic sample types (n = 49). The system can be calibrated with either aqueous standards or leached reference materials.

Characterization of methyl mercury in dental wastewater and correlation with sulfate-reducing bacterial DNA.

Dental wastewater (DWW) was collected over two months from a 12-chair clinic and a single-chair office to identify conditions that may affect Hg methylation. DWW was settled for 24 h and samples were collected from the top and bottom of the supernatant to simulate a range of particles that may escape in-line traps. Total Hg spanned 5 orders of magnitude (0.02-5000 microM), following a log-normal distribution with p10, p50, and p90 concentration values of 0.24, 31 and 4000 microM, respectively; typically well in excess of free aqueous Hg solubility. Methyl Hg was present in high levels (2-270 nM), also following a log-normal distribution with p10, p50, and p90 concentration values of 2.8, 17, and 100 nM, respectively. There were no statistically significant differences (90% CI) in p50 methyl Hg or total Hg between the clinic and office. Methyl Hg was predicted from total Hg data by (+/- 95% CI): Log (Me-Hg) = 0.33 (+/- 0.06) x Log (T-Hg) - 2.27 (+/- 0.13). Total methyl Hg from DWW to U.S. wastewater collection systems is estimated to be 2-5 kg yr(-1). Equilibrium speciation modeling predicted that DWW Hg was primarily in sulfide-Hg complexes, except at high total Hg levels where organo-Hg complexes become significant. DNA extracts amplified by quantitative polymerase chain reaction with primers for total eubacteria and sulfate-reducing bacteria (SRB) indicated that the total eubacterial DNA was composed primarily of SRB, and highly significant correlations were found between methyl Hg and both amplified Desulfobacteraceae (p < 0.0001) and Desulfovibrionacaea DNA (p < 0.00001). Both are known Hg methylators. In marked contrast, there was no significant correlation between methyl Hg and amplified Desulfobulbus DNA, a genus generally not known to methylate Hg at high rates. These results strongly suggest that SRB are implicated in DWW Hg methylation.

Determination of MeHg in environmental sample matrices using Hg-thiourea complex ion chromatography with on-line cold vapor generation and atomic fluorescence spectrometric detection.

A novel system for mercury speciation analysis using high-pressure ion chromatography (IC) has been developed and validated. Its chemistry permits separation of the two most abundant forms of Hg in natural waters, soils, sediments, and biota-monomethyl Hg (CH3Hg+) and mercuric Hg (Hg2+)-on the basis of the difference in charge of their respective thiourea (S=C(NH2)20) complexes. Once separated, both species are converted to Hg0 on-line and quantified by cold-vapor atomic fluorescence spectrometry (CVAFS). A column containing thiol-functionalized silica resin installed in the sample loop of the IC system traps Hg2+ and CH3Hg+ from prepared sample solutions without retaining interfering sample matrix components. The resulting matrix-independent chemistry permits external calibration of the system and a high sample throughput ( approximately 6 samples per hour). The system's accuracy has been validated with environmentally relevant reference materials. Figures of merit for the system, an average precision of approximately 2.5% and an absolute detection limit of <1 pg, are comparable to state-of-the-art gas chromatography approaches for Hg speciation analysis (ethylation/GC-CVAFS) and meet or exceed those of all extant LC systems. Low relative detection limits are attainable through preconcentration onto the thiol resin.