Transcriptomics of Long-Term Meditation Practice: Evidence for Prevention or Reversal of Stress Effects Harmful to Health.

Background and Objectives: Stress can overload adaptive mechanisms, leading to epigenetic effects harmful to health. Research on the reversal of these effects is in its infancy. Early results suggest some meditation techniques have health benefits that grow with repeated practice. This study focused on possible transcriptomic effects of 38 years of twice-daily Transcendental Meditation® (TM®) practice. Materials and Methods: First, using Illumina® BeadChip microarray technology, differences in global gene expression in peripheral blood mononuclear cells (PBMCs) were sought between healthy practitioners and tightly matched controls (n = 12, age 65). Second, these microarray results were verified on a subset of genes using quantitative polymerase chain reaction (qPCR) and were validated using qPCR in larger TM and control groups (n = 45, age 63). Bioinformatics investigation employed Ingenuity® Pathway Analysis (IPA®), DAVID, Genomatix, and R packages. Results: The 200 genes and loci found to meet strict criteria for differential expression in the microarray experiment showed contrasting patterns of expression that distinguished the two groups. Differential expression relating to immune function and energy efficiency were most apparent. In the TM group, relative to the control, all 49 genes associated with inflammation were downregulated, while genes associated with antiviral and antibody components of the defense response were upregulated. The largest expression differences were shown by six genes related to erythrocyte function that appeared to reflect a condition of lower energy efficiency in the control group. Results supporting these gene expression differences were obtained with qPCR-measured expression both in the well-matched microarray groups and in the larger, less well-matched groups. Conclusions: These findings are consistent with predictions based on results from earlier randomized trials of meditation and may provide evidence for stress-related molecular mechanisms underlying reductions in anxiety, post-traumatic stress disorder (PTSD), cardiovascular disease (CVD), and other chronic disorders and diseases.

Organic diet intervention significantly reduces urinary glyphosate levels in U.S. children and adults.

BACKGROUND: A growing set of studies show that an organic diet is associated with reduced levels of urinary pesticide analytes. However, with the exception of one pilot study of two individuals, diet intervention studies to date have not analyzed glyphosate, the most commonly used herbicide in the United States and globally. OBJECTIVE: To investigate the impact of an organic diet intervention on levels of glyphosate and its main metabolite, AMPA (aminomethyl phosphonic acid), in urine collected from adults and children. METHODS: We analyzed urine samples from four racially and geographically diverse families in the United States for five days on a completely non-organic diet and for five days on a completely organic diet (n = 16 participants and a total of 158 urine samples). RESULTS: Mean urinary glyphosate levels for all subjects decreased 70.93% (95% CI -77.96, -61.65, p<0.010) while mean AMPA levels decreased by 76.71% (95% CI -81.54, -70.62, p < 0.010) within six days on an organic diet. Similar decreases in urinary levels of glyphosate and AMPA were observed when data for adults were examined alone, 71.59% (95% CI -82.87, -52.86, p < 0.01) and 83.53% (95% CI -88.42, -76.56, p < 0.01) and when data for children were examined alone, 70.85% (95% CI -78.52, -60.42, p < 0.01) and 69.85% (95% CI -77.56, -59.48, p < 0.01). CONCLUSION: An organic diet was associated with significantly reduced urinary levels of glyphosate and AMPA. The reduction in glyphosate and AMPA levels was rapid, dropping to baseline within three days. This study demonstrates that diet is a primary source of glyphosate exposure and that shifting to an organic diet is an effective way to reduce body burden of glyphosate and its main metabolite, AMPA. This research adds to a growing body of literature indicating that an organic diet may reduce exposure to a range of pesticides in children and adults.

Full-Spectrum Analysis of Bioactive Compounds in Rosemary (Rosmarinus officinalis L.) as Influenced by Different Extraction Methods.

Rosmarinus officinalis is a potent antioxidant herb rich in polyphenols. Ultra-high-performance liquid chromatography, coupled with electrospray ionization and quadrupole-time of flight mass spectrometry (UHPLC-ESI-QTOF-MS), enables an exhaustive, full-spectrum analysis of the molecular constituents of natural products. The study aimed to develop a rapid UHPLC method to contribute new insights into the phytochemical composition of rosemary and to assess the performance of nine different procedures for extraction. These include fresh tissue homogenization, fresh and dry leaf decoction, and their respective fermentation, Soxhlet extraction, and sonication using water and methanol. Different extraction methods were found to recover quite different groups of polyphenols within 11 min during 20 min of analysis. Soxhlet extraction, yielded very high concentrations of rosmarinic acid (33,491.33 ± 86.29 µg/g), luteolin-7-O-glucoside (209.95 ± 8.78 µg/g), carnosic acid (2915.40 ± 33.23 µg/g), carnosol (22,000.67 ± 77.39 µg/g), and ursolic acid (5144.27 ± 28.68 µg/g). UHPLC-ESI-QTOF-MS enabled the detection of more than 50 polyphenols, including phenolic acids, flavonoids, and terpenoids in the various extracts. Of these, sagerinic acid ([M - H]-m/z 719.16), salvianolic acid A ([M - H]-m/z 493.11) and B ([M - H]-m/z 717.15), and a pentacyclic triterpenoid corosolic acid ([M - H]-m/z 471.34) were detected for the first time in rosemary. Soxhlet extraction was found to be the most efficient method, followed by dry leaf decoction. The UHPLC-ESI-QTOF-MS methodology for the analysis proved to be very efficient in the identification and characterization of targeted and untargeted bioactive molecules in the rosemary.

Flock health survey of Irish Texel society breeders and larynx examination in Texel sheep.

BACKGROUND: Laryngeal chondritis is a disease of undetermined aetiology, characterised by oedema, ulceration, abscessation and necrosis of the laryngeal mucosa and cartilage. The initial aim of the study was to document flock health issues identified by Irish pedigree Texel breeders using a questionnaire survey. Additionally, given the reports of breed predisposition for laryngeal chondritis in Texels, a further aim was to identify if laryngeal problems were perceived as an issue. Work was then conducted to identify if pre-clinical laryngeal mucosal pathology was identifiable in Texel sheep showing no overt clinical signs of respiratory disease and if associations existed between laryngeal measurements and laryngeal pathology.Thirty one larynges were collected from a Texel flock that previously had laryngeal chondritis diagnosed in fallen stock. Gross visual inspection was performed to identify and grade (0-5) laryngeal pathology. A series of measurements were then performed on larynges that had been formalin fixed. Associations between independent variables (larynx measurements) and the dependent variable (laryngeal pathology score) were examined. RESULTS: Respiratory disease was the most frequently identified health issue. Farmer-diagnosed 'throat problems' were reported by over 80% of respondents.Laryngeal pathology was noted in Texels showing no overt clinical signs of respiratory disease. Associations between laryngeal measurements and laryngeal pathology were identified relating to the angle between the cranial point of the cricoid cartilage and the vocal process of the arytenoid cartilage. CONCLUSIONS: Mild laryngeal pathology was noted in animals with no overt clinical signs of respiratory disease. Future research should examine whether significant associations between laryngeal measurements and laryngeal pathology identified in the current study can be measured ante mortem, and whether such ante mortem measurements will allow early identification of sheep at risk of developing laryngeal chondritis.

Spoilage indicator bacteria in farmed Atlantic salmon (Salmo salar) stored on ice for 10 days.

This study investigated the growth of indicator and spoilage bacteria on whole Atlantic salmon (Salmo salar) stored aerobically at 2 °C. On days 0, 2, 3, 6, 8 and 10 microbiological analysis was carried out on inner flesh and outer skin samples as well as outer skin swabs (25 cm2 surface areas). Mesophilic total viable counts (TVCm) on skin, flesh and swab samples increased from 1.9, 1.1 and 2.7 log10 CFUcm2 to 6.0, 5.1 and 5.7 log10 CFU/cm2 after 10 days, respectively. Psychrotrophic counts (TVCp), increased from 2.2, 1.8 and 3.1 log10 CFU/cm2 to 6.2, 5.3 and 5.9 log10 CFU/cm2, for skin, flesh and swab samples respectively. Hydrogen sulphide producing bacteria (HSPB), lactic acid bacteria (LAB), Pseudomonas spp., Brochothrix thermosphacta and Photobacterium spp. grew well with similar growth rates (mean generation times of 17.2-26 h). It was concluded that the shelf-life of salmon at 2 °C was approximately 10 days and that HSPB, LAB, Pseudomonas spp., Br. thermosphacta and Photobacterium spp. may be a better indicator of fish spoilage rather than TVC growth, with a count of 5-6 log10 CFU/cm2 indicating the end of shelf-life.

Epidemiological investigation of a severe rumen fluke outbreak on an Irish dairy farm.

Although the rumen fluke, Calicophoron daubneyi is now very common and widespread throughout Western Europe, reports of clinical cases are still rare. This study explores the epidemiological background to a severe rumen fluke outbreak in 6-month-old heifers on a dairy farm in Ireland. Sequence analysis of the cytochrome oxidase subunit 1 (Cox1) gene of the rumen fluke metacercariae on pasture failed to identify predominant, possibly pathogenic subtypes. However, estimates of metacercarial load indicated that the animals were exposed to a daily dose of about 5334 C. daubneyi metacercariae for a period of 3 weeks resulting in the build-up of very large numbers of immature worms in the small intestine. It is hypothesized that specific environmental conditions may favour this parasite over its competitor, the liver fluke, Fasciola hepatica, possibly by allowing it to emerge earlier. The possibility that C. daubneyi may be better adapted to the Irish climate than F. hepatica together with the fact that selective treatment against F. hepatica effectively frees the niche for C. daubneyi, may result in the gradual replacement of F. hepatica by C. daubneyi.

Prevalence of Bovine Viral Diarrhoea Virus (BVDV), Bovine Herpes Virus 1 (BHV 1), Leptospirosis and Neosporosis, and associated risk factors in 161 Irish beef herds.

BACKGROUND: There are limited data available, in Ireland or elsewhere, to determine the extent of exposure to various endemic diseases among beef cows and factors associated with exposure to causative pathogens. The objectives of this study were to determine the herd and within herd prevalence of Bovine Viral Diarrhoea Virus (BVDV), Bovine Herpes Virus 1 (BHV-1), Leptospirosis and Neosporosis in a large scale study of commercial beef herds on the island of Ireland, and to examine herd level factors associated with exposure to these pathogens in these herds. RESULTS: The average number of cows tested per herd was 35.5 (median 30). Herd level seroprevalence to Bovine Herpesvirus-1(BHV-1), Bovine Viral-Diarrhoea Virus (BVDV), Leptospirosis and Neosporosis was 90%, 100%, 91% and 67%, respectively, while the mean within herd prevalence for the these pathogens was 40%, 77.7%, 65.7% and 5.7%, respectively. The study confirms that the level of seroconversion for the four pathogens of interest increases with herd size. There was also evidence that exposure to one pathogen may increase the risk of exposure to another pathogen. CONCLUSIONS: Herd level seroprevalences were in excess of 90% for BVDV, BHV-1 and Leptosporosis. Larger herds were subject to increased exposure to disease pathogens. This study suggests that exposure to several pathogens may be associated with the further exposure to other pathogens.

Kinetic changes during a six-week minimal footwear and gait-retraining intervention in runners.

An evaluation of a six-week Combined minimal footwear transition and gait-retraining combination vs. gait retraining only on impact characteristics and leg stiffness. Twenty-four trained male runners were randomly assigned to either (1) Minimalist footwear transition Combined with gait-retraining over a six-week period ("Combined" group; n = 12) examined in both footwear, or (2) a gait-retraining group only with no minimalist footwear exposure ("Control"; n = 12). Participants were assessed for loading rate, impact peak, vertical, knee and ankle stiffness, and foot-strike using 3D and kinetic analysis. Loading rate was significantly higher in the Combined group in minimal shoes in pre-tests compared to a Control (P ≤ 0.001), reduced significantly in the Combined group over time (P ≤ 0.001), and was not different to the Control group in post-tests (P = 0.16). The impact peak (P = 0.056) and ankle stiffness reduced in both groups (P = 0.006). Loading rate and vertical stiffness was higher in minimalist footwear than conventional running shoes both pre (P ≤ 0.001) and post (P = 0.046) the intervention. There has a higher tendency to non-rearfoot strike in both interventions, but more acute changes in the minimalist footwear. A Combined intervention can potentially reduce impact variables. However, higher loading rate initially in minimalist footwear may increase the risk of injury in this condition.

Comparison of rapid laboratory tests for failure of passive transfer in the bovine.

BACKGROUND: Failure of passive transfer of maternal immunity via colostrum can occur in the bovine, and a number of blood tests have been developed to test calves for this failure. It is not clear which test is most suitable for this purpose. The objective was to examine the most commonly used tests for failure of passive transfer and to decide which is most suitable for routine laboratory use. 126 serum samples were taken from calves of dairy cows after birth but prior to colostrum feeding, and at 48 h of age. Five different tests were compared against radial immunodiffusion which is considered the appropriate reference method. These tests were serum gamma-glutamyltransferase levels, serum protein levels, serum globulin levels, an enzyme linked immunosorbent assay and the zinc sulphate turbidity test. RESULTS: The tests examined displayed high sensitivity but widely varying specificity. Examination of the use of different cut-off points allowed some improvement in specificity at the expense of sensitivity, but the tests which had performed best at the original cut-off points still displayed the best performance. Gamma-glutamyltransferase levels as a measure of colostrum absorption returned, in this study, the best balance between sensitivity and specificity. The ELISA used in this study and serum globulin levels displayed performance similar to the gamma-glutamyltransferase levels. Serum total protein was less successful than others examined at providing both sensitivity and specificity but may, when performed via refractometer, be useful for on-farm testing. As currently performed the poor sensitivity for which the zinc sulphate turbidity test is most often criticized is evident. Modification of the cut-off point to increase specificity is less successful at balancing these parameters than the ELISA, gamma-glutamyltransferase levels, and globulin levels. CONCLUSIONS: Gamma-glutamyltransferase levels, ELISA testing and circulating globulin levels performed best in detecting failure of passive transfer in serum samples, although all three had some practical considerations.

Corrigendum: Assessment of a novel Ehlers-Danlos syndromes disability index.

[This corrects the article DOI: 10.3389/fresc.2024.1280582.].

Assessment of a novel Ehlers-Danlos syndromes disability index.

Background: The Ehlers-Danlos syndromes (EDS) are a group of inherited connective tissue disorders characterized by disruptions in collagen synthesis and processing. These disorders lead to various symptoms, including hypermobility, musculoskeletal conditions, and chronic pain that can significantly limit patients' daily living. In the absence of a curative treatment, an EDS specific disability index that tracks changes in patient-reported outcomes can facilitate the investigation of new treatment options and enhance the quality of life for EDS patients. Methods: An EDS-specific disability index was created using survey data and input from clinicians. A total of 222 EDS patients in a multidisciplinary clinical program completed the index during their initial visit. Exploratory and confirmatory factor analyses were conducted to determine the index's factor solution and assess its goodness-of-fit. Paired t-tests were performed with follow-up visit data collected over the course of one year. Results: The exploratory and confirmatory factor analyses indicated a two-factor solution, accounting for 42.40% of the variance. The index demonstrated adequate fit to the data, supported by Tucker and Lewis's index (0.85) and root mean square error of approximation (0.1). Data from follow-up visits showed significant improvement in three symptom related variables and one function related variable in addition to the total score and the symptom subscale score when compared to the initial visit. Conclusion: The development of an EDS-specific disability index is a crucial step in creating a clinical tool that enables healthcare professionals to gain a deeper understanding of the impact EDS has on patients' lives and potentially identify new therapeutic interventions.

Paleoamerican exploitation of extinct megafauna revealed through immunological blood residue and microwear analysis, North and South Carolina, USA.

Previous immunological studies in the eastern USA have failed to establish a direct connection between Paleoamericans and extinct megafauna species. The lack of physical evidence for the presence of extinct megafauna begs the question, did early Paleoamericans regularly hunt or scavenge these animals, or were some megafauna already extinct? In this study of 120 Paleoamerican stone tools from across North and South Carolina, we investigate this question using crossover immunoelectrophoresis (CIEP). We find immunological support for the exploitation of extant and extinct megafauna, including Proboscidea, Equidae, and Bovidae (possibly Bison antiquus), on Clovis points and scrapers, as well as possible early Paleoamerican Haw River points. Post-Clovis points tested positive for Equidae and Bovidae but not Proboscidea. Microwear results are consistent with projectile usage, butchery, fresh- and dry hide scraping, the use of ochre-coated dry hides for hafting, and dry hide sheath wear. This study represents the first direct evidence of the exploitation of extinct megafauna by Clovis and other Paleoamerican cultures in the Carolinas and more broadly, across the eastern United States, where there is generally poor to non-existent faunal preservation. Future CIEP analysis of stone tools may provide evidence on the timing and demography of megafaunal collapse leading to eventual extinction.

Correction: Chhalliyil et al. A Real-Time Quantitative PCR Method Specific for Detection and Quantification of the First Commercialized Genome-Edited Plant. Foods 2020, 9, 1245.

The stated objective of the paper was to provide regulatory laboratories and industry laboratories a complete, legally robust method for the quantitative detection of SU canola [...].

High-Pressure Processing for the Production of Added-Value Claw Meat from Edible Crab (Cancer pagurus).

High-pressure processing (HPP) in a large-scale industrial unit was explored as a means for producing added-value claw meat products from edible crab (Cancer pagurus). Quality attributes were comparatively evaluated on the meat extracted from pressurized (300 MPa/2 min, 300 MPa/4 min, 500 MPa/2 min) or cooked (92 °C/15 min) chelipeds (i.e., the limb bearing the claw), before and after a thermal in-pack pasteurization (F9010 = 10). Satisfactory meat detachment from the shell was achieved due to HPP-induced cold protein denaturation. Compared to cooked or cooked-pasteurized counterparts, pressurized claws showed significantly higher yield (p < 0.05), which was possibly related to higher intra-myofibrillar water as evidenced by relaxometry data, together with lower volatile nitrogen levels. The polyunsaturated fatty acids content was unaffected, whereas the inactivation of total viable psychrotrophic and mesophilic bacteria increased with treatment pressure and time (1.1-1.9 log10 CFU g-1). Notably, pressurization at 300 MPa for 4 min resulted in meat with no discolorations and, after pasteurization, with high color similarity (ΔE* = 1.2-1.9) to conventionally thermally processed samples. Following further investigations into eating quality and microbiological stability, these HPP conditions could be exploited for producing uncooked ready-to-heat or pasteurized ready-to-eat claw meat products from edible crab.

A Real-Time Quantitative PCR Method Specific for Detection and Quantification of the First Commercialized Genome-Edited Plant.

Discussion regarding the regulatory status of genome-edited crops has focused on precision of editing and on doubts regarding the feasibility of analytical monitoring compliant with existing GMO regulations. Effective detection methods are important, both for regulatory enforcement and traceability in case of biosafety, environmental or socio-economic impacts. Here, we approach the analysis question for the first time in the laboratory and report the successful development of a quantitative PCR detection method for the first commercialized genome-edited crop, a canola with a single base pair edit conferring herbicide tolerance. The method is highly sensitive and specific (quantification limit, 0.05%), compatible with the standards of practice, equipment and expertise typical in GMO laboratories, and readily integrable into their analytical workflows, including use of the matrix approach. The method, validated by an independent laboratory, meets all legal requirements for GMO analytical methods in jurisdictions such as the EU, is consistent with ISO17025 accreditation standards and has been placed in the public domain. Having developed a qPCR method for the most challenging class of genome edits, single-nucleotide variants, this research suggests that qPCR-based method development may be applicable to virtually any genome-edited organism. This advance resolves doubts regarding the feasibility of extending the regulatory approach currently employed for recombinant DNA-based GMOs to genome-edited organisms.

Descriptive analysis of ovine mortality in sentinel sheep flocks in Ireland.

BACKGROUND: Studies of sheep mortality or cause-specific mortality, in Ireland or internationally, are relatively scarce but are important in presenting baseline levels and changing trends of endemic disease. This study assessed sheep mortality and cause-specific mortality in 33 sentinel sheep flocks in Ireland. METHODS: Sentinel flocks were requested to submit carcases of all sheep that died to the regional veterinary laboratories (RVLs) of the Department of Agriculture, Food and Marine during a calendar year (2016). Postmortem examinations were performed on 1247 submissions to Athlone, Kilkenny and Sligo RVLs. RESULTS: The median overall submission rate was 13.8 per cent (range 2.5 per cent-35.8 per cent) per adult female sheep in the flock in January 2016. The median fetal, perinatal, lamb and adult submissions per adult female sheep in the flock in January 2016 were 2.1 per cent (0.0 per cent-15.2 per cent), 3.5 per cent (0.0 per cent-20.0 per cent), 3.0 per cent (0.0 per cent-12.4 per cent) and 2.8 per cent (0.8 per cent-7.1 per cent), respectively. The frequency of detection of categories of postmortem diagnoses in fetuses, perinates, lambs and adults are presented. CONCLUSIONS: Comparisons with existing passive surveillance findings reflect some differences in the relative frequency of detection of certain categories of disease suggesting that sentinel flock surveillance could usefully supplement existing passive animal disease surveillance activities for ovine disease.

Real-time and conventional PCR detection of Liberty Link rice varieties and transgenic soy in rice sampled in the Mexican and American retail markets.

Samples of rice from Mexican and USA retail stores were analyzed for the presence of transgenic (GM) events using real-time PCR. In screening for the CaMV35S promoter sequence (35SP), positive results were found in 49 and 35% of the Mexican and American samples, respectively. In further investigations in Mexican samples, 43% were positive for P35S::bar, with two above the quantifiable limit; these were 0.07% and 0.05% GMO. Fourteen out of the sixteen positive samples were labeled as imported from the USA. In testing samples bought in American retail shops, 24% showed positive results, all below the quantifiable range. It could be deduced that P35S::bar positive samples were Liberty Link(R) (LL) rice. In distinguishing between LL601 and LL62, end-point PCR was used, corroborating the P35S::bar amplicon length difference of these events. LL62 was found in one rice sample purchased in Mexico and two in the USA samples. Its presence was verified with the 35S terminator sequence. All other LL positive samples contained LL601. None of the samples analyzed showed the presence of Bt63 rice. The LL rice varieties found have been identified as not being commercially cultivated, and so their presence requires further investigation. 35SP was also present in samples which did not have any LL rice. Maize sequences could not be detected in any of the samples; however, soybean DNA was found in Mexican and USA rice samples. The Roundup Ready(R) trait was detected in trace amounts in 16 and 6% of the rice samples bought in Mexico and the USA, respectively. Real-time PCR was shown to be the method of choice for the sensitive and rapid screening of commodities and retail samples for the detection of GM and other contamination.

Treacher Collins syndrome: an undescribed characteristic of the condition and its management with botulinum toxin and surgery.

Mandibulofacial dysostosis (Treacher Collins syndrome) is associated with clinical abnormalities of structures derived from the first and second branchial arches, including antimongoloid slant of palpebral fissures, colobomas of the lower eyelid, eyelash malformations, and malar and mandibular defects. We describe an unusual clinical feature associated with colobomas of the lower eyelids, in a patient with mandibulofacial dysostosis, successfully treated with botulinum toxin and subsequent surgery.