Single Nucleotide Polymorphism Directed Antiemetic Treatment in Women With Breast Cancer Treated With Neo- or Adjuvant Chemotherapy: A Randomised Multicentre Phase II Study. (EudraCT: 2015-000658-39).

BACKGROUND/AIM: The role of single nucleotide polymorphisms (SNPs) in the frequency and intensity of chemotherapy-induced nausea and vomiting (CINV) in women with breast cancer (BC) is unclear. The primary purpose of this study was to compare/evaluate the effect of SNP-guided antiemetic treatment versus standard CINV treatment. PATIENTS AND METHODS: A randomised, factorial, phase II multicentre study design was used. Women planned for neoadjuvant or adjuvant chemotherapy with epirubicin, cyclophosphamide and fluorouracil (FEC /EC, with or without fluorouracil) for BC were randomised to SNP-guided antiemetic treatment (based on the results of SNP analyses) versus standard CINV treatment. Blood samples were taken before the treatment was initiated. Patient-reported data on CINV (during 10 days from onset of cancer treatment) and health-related quality of life (HRQoL), were collected before and after the first cancer treatment. RESULTS: A total of 188 women were included. Overall, nausea was reported by 86% (n=129) of the patients during the ten-day period from the start of cancer treatment. The SNP genotype studied varied. In FAS-CD95, the genotypes AG and GG were overrepresented; in RB1-LPAR6, GG was overrepresented, and in CCL2, both AA and GG were overrepresented. We found no statistically significant difference in CINV between SNP-guided antiemetic treatment versus standard CINV treatment. CONCLUSION: SNP-guided antiemetic treatment could be as effective as standard treatment. SNP-guided antiemetic treatment of CINV is possibly useful in detecting patients with a higher or lower risk for CINV and thus may help in avoiding over-treatment with toxic components. CINV negatively affects the HRQL.

A Multi-Ingredient Supplement Protects against Obesity and Infertility in Western Diet-Fed Mice.

The Western diet (WD) predisposes to bodyweight gain and obesity and is linked to mitochondrial dysfunction, oxidative damage, inflammation, and multisystem disease, even affecting the reproductive organs, fertility, and pregnancy outcomes. In this study, we investigated the effects of multi-ingredient supplementation (MIS) with antioxidants, phytonutrients, and vitamins ('Fertility Enhancer'; FE) on white adipose tissue (WAT) expansion, nonalcoholic fatty liver disease (NAFLD), and infertility in WD-fed C57BL/6J mice. Five-month-old male (M) and female (F) mice were fed a low-fat diet (LF) or a high fat/sucrose WD (HF) for six weeks, followed by six weeks of LF (3.64 kcal/g), HF (4.56 kcal/g), or HF combined with FE (4.50 kcal/g). A sub-set of animals were sacrificed at 12 weeks, while the remainder were harem-mated in a 1:2 male-to-female ratio, and singly housed during the gestational period. Two-way, factorial ANOVA analysis revealed a main effect of diet on bodyweight (BW), total body fat, % body fat, white adipose tissue mass, and liver lipid content (all p < 0.001), driven by the anti-obesogenic effects of the 'Fertility Enhancer'. Similarly, a main effect of diet was found on PGC1-α mRNA levels (p < 0.05) and mitochondrial protein content (p < 0.001) in perigonadal WAT, with PGC1-α induction and higher complex II and complex III expression in FE vs. HF animals. Copulatory plug counts were higher in FE vs. HE couples (30% vs. 6%), resulting in more litters (4 vs. 0) and higher copulatory success (67% vs. 0%). Although the trends of all histology outcomes were suggestive of a benefit from the FE diet, only the number of atretic follicles and testicular mass were significant. Ovarian IL-1ß mRNA induction was significantly attenuated in the FE group (p < 0.05 vs. HF) with CASP1 attenuation trending lower (p = 0.09 vs. HF), which is indicative of anti-inflammatory benefits of the 'Fertility Enhancer.' We conclude that supplementation with specific phytonutrients, antioxidants, and vitamins may have utility as an adjunctive therapy for weight management, fatty liver disease, and infertility in overweight and obese couples.

Nutritional co-therapy with 1,3-butanediol and multi-ingredient antioxidants enhances autophagic clearance in Pompe disease.

Alglucosidase alpha is an orphan drug approved for enzyme replacement therapy (ERT) in Pompe disease (PD); however, its efficacy is limited in skeletal muscle because of a partial blockage of autophagic flux that hinders intracellular trafficking and enzyme delivery. Adjunctive therapies that enhance autophagic flux and protect mitochondrial integrity may alleviate autophagic blockage and oxidative stress and thereby improve ERT efficacy in PD. In this study, we compared the benefits of ERT combined with a ketogenic diet (ERT-KETO), daily administration of an oral ketone precursor (1,3-butanediol; ERT-BD), a multi-ingredient antioxidant diet (ERT-MITO; CoQ10, α-lipoic acid, vitamin E, beetroot extract, HMB, creatine, and citrulline), or co-therapy with the ketone precursor and multi-ingredient antioxidants (ERT-BD-MITO) on skeletal muscle pathology in GAA-KO mice. We found that two months of 1,3-BD administration raised circulatory ketone levels to ≥1.2 mM, attenuated autophagic buildup in type 2 muscle fibers, and preserved muscle strength and function in ERT-treated GAA-KO mice. Collectively, ERT-BD was more effective vs. standard ERT and ERT-KETO in terms of autophagic clearance, dampening of oxidative stress, and muscle maintenance. However, the addition of multi-ingredient antioxidants (ERT-BD-MITO) provided the most consistent benefits across all outcome measures and normalized mitochondrial protein expression in GAA-KO mice. We therefore conclude that nutritional co-therapy with 1,3-butanediol and multi-ingredient antioxidants may provide an alternative to ketogenic diets for inducing ketosis and enhancing autophagic flux in PD patients.

A Novel Multi-Ingredient Supplement Activates a Browning Program in White Adipose Tissue and Mitigates Weight Gain in High-Fat Diet-Fed Mice.

We investigated the effects of a novel multi-ingredient supplement comprised of polyphenol antioxidants and compounds known to facilitate mitochondrial function and metabolic enhancement (ME) in a mouse model of obesity. In this study, 6-week-old male C57/BL6J mice were placed on a high-fat diet (HFD; ~60% fat) for 6 weeks, with subsequent allocation into experimentalgroups for 4 weeks: HFD control, HFD + ME10 (10 components), HFD + ME7 (7 components), HFD + ME10 + EX, HFD + EX (where '+EX' animals exercised 3 days/week), and chow-fed control. After the intervention, HFD control animals had significantly greater body weight and fat mass. Despite the continuation of HFD, animals supplemented with multi-ingredient ME or who performed exercise training showed an attenuation of fat mass and preservation of lean body mass, which was further enhanced when combined (ME+EX). ME supplementation stimulated the upregulation of white and brown adipose tissue mRNA transcripts associated with mitochondrial biogenesis, browning, fatty acid transport, and fat metabolism. In WAT depots, this was mirrored by mitochodrial oxidative phosphorylation (OXPHOS) protein expression, and increased in vivo fat oxidation measured via CLAMS. ME supplementation also decreased systemic and local inflammation markers. Herein, we demonstrated that novel multi-ingredient nutritional supplements induced significant fat loss independent of physical activity while preserving muscle mass in obese mice. Mechanistically, these MEs appear to act by inducing a browning program in white adipose tissue and decreasing other pathophysiological impairments associated with obesity, including mitochondrial respiration alterations induced by HFD.

A Five-Ingredient Nutritional Supplement and Home-Based Resistance Exercise Improve Lean Mass and Strength in Free-Living Elderly.

Old age is associated with lower physical activity levels, suboptimal protein intake, and desensitization to anabolic stimuli, predisposing for age-related muscle loss (sarcopenia). Although resistance exercise (RE) and protein supplementation partially protect against sarcopenia under controlled conditions, the efficacy of home-based, unsupervised RE (HBRE) and multi-ingredient supplementation (MIS) is largely unknown. In this randomized, placebo-controlled and double-blind trial, we examined the effects of HBRE/MIS on muscle mass, strength, and function in free-living, older men. Thirty-two sedentary men underwent twelve weeks of home-based resistance band training (3 d/week), in combination with daily intake of a novel five-nutrient supplement ('Muscle5'; M5, n = 16, 77.4 ± 2.8 y) containing whey, micellar casein, creatine, vitamin D, and omega-3 fatty acids, or an isocaloric/isonitrogenous placebo (PLA; n = 16, 74.4 ± 1.3 y), containing collagen and sunflower oil. Appendicular and total lean mass (ASM; +3%, TLM; +2%), lean mass to fat ratios (ASM/% body fat; +6%, TLM/% body fat; +5%), maximal strength (grip; +8%, leg press; +17%), and function (5-Times Sit-to-Stand time; -9%) were significantly improved in the M5 group following HBRE/MIS therapy (pre vs. post tests; p < 0.05). Fast-twitch muscle fiber cross-sectional areas of the quadriceps muscle were also significantly increased in the M5 group post intervention (Type IIa; +30.9%, Type IIx, +28.5%, p < 0.05). Sub-group analysis indicated even greater gains in total lean mass in sarcopenic individuals following HBRE/MIS therapy (TLM; +1.65 kg/+3.4%, p < 0.05). We conclude that the Muscle5 supplement is a safe, well-tolerated, and effective complement to low-intensity, home-based resistance exercise and improves lean mass, strength, and overall muscle quality in old age.

Nutrition and exercise in Pompe disease.

The current standard of care for Pompe disease (PD) is the administration of enzyme replacement therapy (ERT). Exercise and nutrition are often considered as complementary strategies rather than "treatments" per se. Nutritional assessment is important in patients with locomotor disability because the relative hypodynamia limits energy expenditure and thus the total amount of energy must be reduced to avoid obesity. A lower total energy intake often leads to lower protein and micronutrient intake. Consequently, ensuring that Pompe patients are tested for and replaced for deficiencies (protein, vitamin D, vitamin B12, etc.) is an important aspect of care. Furthermore, given the role of autophagy in the pathophysiology of PD and the fact that fasting induces autophagy, it is important that strategies such as nutritional timing and amino acid intake (L-arginine, L-leucine) be evaluated as therapies. Exercise interventions have been shown to improve six-minute walk testing distance by more than what was seen in the seminal ERT study in late-onset PD. Exercise therapy can also activate autophagy, and this is likely another component of its efficacy. The current review will evaluate the theoretical and practical aspects of nutrition and exercise as therapies for patients with PD.

Mixed support for a causal link between single dose intranasal oxytocin and spiritual experiences: opposing effects depending on individual proclivities for absorption.

Intranasal oxytocin (OT) has previously been found to increase spirituality, an effect moderated by OT-related genotypes. This pre-registered study sought to conceptually replicate and extend those findings. Using a single dose of intranasal OT vs placebo (PL), we investigated experimental treatment effects, and moderation by OT-related genotypes on spirituality, mystical experiences, and the sensed presence of a sentient being. A more exploratory aim was to test for interactions between treatment and the personality disposition absorption on these spirituality-related outcomes. A priming plus sensory deprivation procedure that has facilitated spiritual experiences in previous studies was used. The sample (N = 116) contained both sexes and was drawn from a relatively secular context. Results failed to conceptually replicate both the main effects of treatment and the treatment by genotype interactions on spirituality. Similarly, there were no such effects on mystical experiences or sensed presence. However, the data suggested an interaction between treatment and absorption. Relative to PL, OT seemed to enhance spiritual experiences in participants scoring low in absorption and dampen spirituality in participants scoring high in absorption.

Limited reverse transcriptase activity of phi29 DNA polymerase.

Phi29 (Φ29) DNA polymerase is an enzyme commonly used in DNA amplification methods such as rolling circle amplification (RCA) and multiple strand displacement amplification (MDA), as well as in DNA sequencing methods such as single molecule real time (SMRT) sequencing. Here, we report the ability of phi29 DNA polymerase to amplify RNA-containing circular substrates during RCA. We found that circular substrates with single RNA substitutions are amplified at a similar amplification rate as non-chimeric DNA substrates, and that consecutive RNA pyrimidines were generally preferred over purines. We observed RCA suppression with higher number of ribonucleotide substitutions, which was partially restored by interspacing RNA bases with DNA. We show that supplementing manganese ions as cofactor supports replication of RNAs during RCA. Sequencing of the RCA products demonstrated accurate base incorporation at the RNA base with both Mn2+ and Mg2+ as cofactors during replication, proving reverse transcriptase activity of the phi29 DNA polymerase. In summary, the ability of phi29 DNA polymerase to accept RNA-containing substrates broadens the spectrum of applications for phi29 DNA polymerase-mediated RCA. These include amplification of chimeric circular probes, such as padlock probes and molecular inversion probes.

Visual Task Demands and the Auditory Mismatch Negativity: An Empirical Study and a Meta-Analysis.

Because the auditory system is particularly useful in monitoring the environment, previous research has examined whether task-irrelevant, auditory distracters are processed even if subjects focus their attention on visual stimuli. This research suggests that attentionally demanding visual tasks decrease the auditory mismatch negativity (MMN) to simultaneously presented auditory distractors. Because a recent behavioral study found that high visual perceptual load decreased detection sensitivity of simultaneous tones, we used a similar task (n = 28) to determine if high visual perceptual load would reduce the auditory MMN. Results suggested that perceptual load did not decrease the MMN. At face value, these nonsignificant findings may suggest that effects of perceptual load on the MMN are smaller than those of other demanding visual tasks. If so, effect sizes should differ systematically between the present and previous studies. We conducted a selective meta-analysis of published studies in which the MMN was derived from the EEG, the visual task demands were continuous and varied between high and low within the same task, and the task-irrelevant tones were presented in a typical oddball paradigm simultaneously with the visual stimuli. Because the meta-analysis suggested that the present (null) findings did not differ systematically from previous findings, the available evidence was combined. Results of this meta-analysis confirmed that demanding visual tasks reduce the MMN to auditory distracters. However, because the meta-analysis was based on small studies and because of the risk for publication biases, future studies should be preregistered with large samples (n > 150) to provide confirmatory evidence for the results of the present meta-analysis. These future studies should also use control conditions that reduce confounding effects of neural adaptation, and use load manipulations that are defined independently from their effects on the MMN.

Blind people are more sensitive than sighted people to binaural sound-location cues, particularly inter-aural level differences.

Blind people use auditory information to locate sound sources and sound-reflecting objects (echolocation). Sound source localization benefits from the hearing system's ability to suppress distracting sound reflections, whereas echolocation would benefit from "unsuppressing" these reflections. To clarify how these potentially conflicting aspects of spatial hearing interact in blind versus sighted listeners, we measured discrimination thresholds for two binaural location cues: inter-aural level differences (ILDs) and inter-aural time differences (ITDs). The ILDs or ITDs were present in single clicks, in the leading component of click pairs, or in the lagging component of click pairs, exploiting processes related to both sound source localization and echolocation. We tested 23 blind (mean age = 54 y), 23 sighted-age-matched (mean age = 54 y), and 42 sighted-young (mean age = 26 y) listeners. The results suggested greater ILD sensitivity for blind than for sighted listeners. The blind group's superiority was particularly evident for ILD-lag-click discrimination, suggesting not only enhanced ILD sensitivity in general but also increased ability to unsuppress lagging clicks. This may be related to the blind person's experience of localizing reflected sounds, for which ILDs may be more efficient than ITDs. On the ITD-discrimination tasks, the blind listeners performed better than the sighted age-matched listeners, but not better than the sighted young listeners. ITD sensitivity declines with age, and the equal performance of the blind listeners compared to a group of substantially younger listeners is consistent with the notion that blind people's experience may offset age-related decline in ITD sensitivity.

Aircraft noise and speech intelligibility in an outdoor living space.

Studies of effects on speech intelligibility from aircraft noise in outdoor places are currently lacking. To explore these effects, first-order ambisonic recordings of aircraft noise were reproduced outdoors in a pergola. The average background level was 47 dB LA eq. Lists of phonetically balanced words (LAS max,word = 54 dB) were reproduced simultaneously with aircraft passage noise (LAS max,noise = 72-84 dB). Twenty individually tested listeners wrote down each presented word while seated in the pergola. The main results were (i) aircraft noise negatively affects speech intelligibility at sound pressure levels that exceed those of the speech sound (signal-to-noise ratio, S/N < 0), and (ii) the simple A-weighted S/N ratio was nearly as good an indicator of speech intelligibility as were two more advanced models, the Speech Intelligibility Index and Glasberg and Moore's [J. Audio Eng. Soc. 53, 906-918 (2005)] partial loudness model. This suggests that any of these indicators is applicable for predicting effects of aircraft noise on speech intelligibility outdoors.

Asymmetric transfer of sound localization learning between indistinguishable interaural cues.

Learning in perceptual tasks is typically highly specific to the trained stimulus parameters. However, can learning be specific to a stimulus parameter that is perceptually indistinguishable from another? We assessed this question using a perceived sound location task in which the perceived sound location was created through either an interaural time difference (ITD) cue or an interaural level difference (ILD) cue. We used the same transient, broadband sound (clicks) for both cues, and after training on one of the cues, listeners switched cue mid-session. This allowed us to assess cue specificity or transfer when the subjectively unnoticed cue switch occurred. One group of listeners improved their ITD performance as a function of training, but deteriorated in performance when switching to ILD in mid training session. Another group of listeners started with ILD training; their improved performance level did not deteriorate as they switched to the ITD cue. This transfer asymmetry was not hypothesized, and we therefore extended our study with a second data collection. Both the training effect and the transfer asymmetry remained after the second data collection. Our results indicate (a) listeners can improve both their ITD and ILD performance for click sounds, extending previous findings on tones; (b) learning can be specific to a stimulus parameter that is indistinguishable from another, as ITD learning did not transfer to ILD performance; but (c) ILD learning can transfer to ITD performance. This transfer asymmetry may have occurred because of how ITD and ILD are coded in early brainstem areas.

Prevalence of dental erosion and association with lifestyle factors in Swedish 20-year olds.

OBJECTIVE: To investigate the prevalence, distribution and severity of dental erosion and its association with lifestyle, oral and general health in young adults. MATERIALS AND METHODS: Four hundred and ninety-four individuals, 20-years of age, participated. Dental erosion in molars and maxillary incisors was evaluated. Caries, plaque and gingivitis were registered. Saliva samples were taken and the subjects were interviewed about behavioural and dietary habits and oral and general health. Body mass index (BMI) was calculated. The individuals were sub-divided into two groups according to the presence and absence of dental erosion: within the group with erosion was a sub-group of individuals with extensive erosion. RESULTS: Of the individuals 25% had no erosion, 75% had erosion and 18% had extensive erosion. Erosion was found in molars in 74% of the individuals and on buccal and palatal surfaces in maxillary incisors in 4% and 7%, respectively. Cupping was seen in 65% of individuals and severe erosion in molars in 1.6%. Compared to subjects with no erosion, those with extensive erosion had a higher consumption of soft drinks (p = 0.05), caries prevalence (p < 0.01), prevalence of mutans Streptococci (p < 0.01) and BMI (p < 0.05). Furthermore, subjects with erosion had higher caries prevalence (p < 0.01) and BMI (p < 0.01) than those with no erosion. CONCLUSIONS: Swedish young adults have a high prevalence of dental erosion, but the level of severe erosion is low. The study disclosed a relationship between dental erosion and behavioural factors, oral health and BMI.

Frequency, type and factors associated with the use of complementary and alternative medicine in patients with Parkinson's disease at a neurological outpatient clinic.

BACKGROUND: Complementary and alternative medicine (CAM) is described as a set of healthcare and medical practices not currently part of conventional western medicine. In this study, we aimed to investigate the frequency, type and factors associated with the use of CAM in Parkinson's disease (PD) patients at a neurological outpatient clinic. METHOD: All PD patients at the outpatient clinic (N = 421) received a postal questionnaire covering ten types of CAM plus an open alternative labelled 'other type', their possible effect, and socio-demographic, health and disease-related factors. RESULTS: 325/403 (81%) eligible patients responded comprising 172 men and 153 women, median age 74 years for both genders. CAM use was reported by (M/F) 51/59 patients (30/39%); 31/33 patients (18/22%) taking CAM drugs and 48/57 (28/37%) using CAM methods with acupuncture having the highest frequency. Significant differences between CAM users and non-users were found with regard to educational level, perceived health and levodopa load. More females than males used CAM, but the difference was not significant. The majority of CAM users perceived 'No improvement' or 'Some improvement'. Twenty percent of users reported that they had spent the equivalent of 50 euro on CAM during the last 6 months. CONCLUSION: CAM use is rather common among PD outpatients, with one in three having experience of CAM practices. Badly-perceived health and higher education were factors significantly associated with CAM utilization. A trend for more female CAM users was also seen.

Evaluating roadside noise barriers using an annoyance-reduction criterion.

A common indicator of effectiveness for roadside noise barriers is the reduction in A-weighted sound pressure level (L(A)). The present experimental study considered alternative indicators using an annoyance-reduction criterion. A large number of 8 s experimental sounds were created from binaural recordings conducted at various distances from a highway at a location with a 4.6 m high roadside barrier (barrier sounds) and at a location along the same road with no barrier (nonbarrier sounds). Eighteen listeners scaled the annoyance of the experimental sounds with the method of magnitude estimation. The barrier sounds recorded 10-45 m from the road and nonbarrier sounds recorded 50-200 m from the road were of similar L(A). Despite this, the barrier sounds were found to be more annoying than the nonbarrier sounds. The annoyance difference corresponded to approximately a 3 dB increase in L(A) and was mainly related to the barrier sounds' higher relative level of low-frequency sound. This suggests that L(A) reduction may not be a valid indicator of the annoyance reduction caused by a noise barrier. The loudness level (ISO 532B) and a low-frequency corrected sound pressure level (L(A) ( *)) were found to be better than L(A) as indicators of the barrier's annoyance-reduction efficiency.

New proposals for testing drugs with IKr-blocking activity to determine their teratogenic potential.

Drugs blocking the potassium current IKr, either as an intended pharmacologic effect (eg antiarrhythmics dofetilide and almokalant) or as an unwanted side-effect (eg antihistamine astemizole, propulsive drug cisapride, antidepressive drugs and macrolide antibiotics) are potential human teratogens. It is the contention of this paper that the existing repeat dose regimen used in teratology studies to fulfil regulatory requirements, does not properly identify the teratogenic risk of these drugs. Results from conventional studies for dofetilide and almokalant showed high rates of postimplantation embryonic death with few malformed fetuses. For astemizole and cisapride only embryonic death was seen. These latter results were not considered important because they occurred either in the presence of maternal toxicity and/or at high doses. Subsequent studies have shown that IKr-blockers are highly teratogenic when administered on single gestational days (GD) during a sensitive period of rat pregnancy (GD 10-14) when they induce a high incidence of stage-specific malformations. This teratogenic activity of astemizole and cisapride was missed in the original teratology studies. Mechanistically IKr-blockers cause bradycardia and arrhythmia of the embryonic heart and while an embryo may be able to survive a single day exposure to a teratogenic dose, repeat dosing often leads to death of the embryo. With this review we suggest that new drugs identified at the preclinical stage of development as having IKr-blocking properties, should undergo more comprehensive teratology testing including single GD dosing and studies using embryo culture. This would further help identify and characterise their teratogenic potential.

Effects of surface sorption on microbial degradation of glyphosate.

Sorption may affect the bioavailability and biodegradation of pesticides in soils. The aim of this study was to test the effect of surface sorption on microbial utilization of the herbicide glyphosate as a source of phosphorus, nitrogen, or carbon. We added goethite to a humus soil to manipulate the soil's glyphosate sorption capacity. The addition of glyphosate generally either decreased microbial CO2 production or produced no effect. Additions of glyphosate, in combination with glucose and N, did not change the respiration rate in comparison with the same treatment but without glyphosate. In contrast, glyphosate additions combined with glucose and P decreased microbial growth, whereas the combination with goethite counteracted the negative effect. The different treatments were examined using attenuated total reflectance Fourier transform (ATR-FTIR) spectroscopy; the results suggest that glyphosate was de-carboxylated in the sorbed state. Stimulating microbial growth by the addition of glucose and nitrogen resulted in further oxidation of glyphosate and only phosphate was detectable on the goethite surface after 13 days incubation. Our results show that sorbed glyphosate is microbially degradable, and it retards microbial activity. This study emphasizes the importance of combining quantitative measurements with a molecular-level examination, to better understand biogeochemical processes.

Parallel gene analysis with allele-specific padlock probes and tag microarrays.

Parallel, highly specific analysis methods are required to take advantage of the extensive information about DNA sequence variation and of expressed sequences. We present a scalable laboratory technique suitable to analyze numerous target sequences in multiplexed assays. Sets of padlock probes were applied to analyze single nucleotide variation directly in total genomic DNA or cDNA for parallel genotyping or gene expression analysis. All reacted probes were then co-amplified and identified by hybridization to a standard tag oligonucleotide array. The technique was illustrated by analyzing normal and pathogenic variation within the Wilson disease-related ATP7B gene, both at the level of DNA and RNA, using allele-specific padlock probes.