Stable Marker Research Articles

Rapid and ultrasensitive electrochemical detection of DNA methylation for ovarian cancer diagnosis

Alterations in DNA methylation, a stable epigenetic marker, are important components in the development of cancer. It is vital to develop diagnostic systems with the ability to rapidly quantify DNA methylation with high sensitivity and selectivity. However, the analysis of DNA methylation must address two main challenges: (i) ultralow abundance and (ii) differentiating methylated cytosine from normal cytosine on target DNA sequence in the presence of an overwhelming background of circulating cell-free DNA. Here we report the development of an ultrasensitive and highly-selective electrochemical biosensor for the rapid detection of DNA methylation in blood. The sensing of DNA methylation involves the hybridization on a network of probe DNA modified gold-coated magnetic nanoparticles (DNA-Au@MNPs) complementary to target DNA, and subsequently enzymatic cleavage to differentiate methylated DNA strands from corresponding unmethylated DNA strands. The biosensor presents a dynamic range from 2 aM to 20 nM for 110 nucleotide DNA sequences containing a single-site methylation with the lowest detected concentration of 2 aM. This DNA-Au@MNPs based sensor provides a promising method to achieve 35 min response time and minimally invasive diagnosis of ovarian cancer.

Simultaneous quantification by LC/ESI–MS/MS of chlorinated tyrosine derivatives in the autopsy sample of a victim of chlorine exposure

Direct detection and accurate quantification of chlorine in autopsy samples are difficult because of the volatility and rapid metabolism of chlorine. Here, we developed and validated a method for quantitative analysis of 3-chloro-l-tyrosine (Cl-Tyr) and 3,5-dichloro-l-tyrosine (DiCl-Tyr) as stable markers of chlorine exposure. Chemical derivatization followed by liquid chromatography coupled with electrospray ionization–tandem mass spectrometry (LC/ESI–MS/MS) enabled us to simultaneously analyze both Cl-Tyr and DiCl-Tyr in an autopsy sample from the victim of chlorine exposure. Cl-Tyr was detected in the heart blood (53.6 ng/mL), urine (9.5 ng/mL), and lung tissue (211.1 ng/g); however, DiCl-Tyr was detected only in the lung tissue (10.3 ng/g). In contrast, in autopsy samples obtained from cases without exposure to chlorine, DiCl-Tyr was not detected in any matrixes. Our result suggested that the simultaneous detection of Cl-Tyr and DiCl-Ty may provide a better appreciation of chlorine exposure. To our knowledge, this is the first time Cl-Tyr and DiCl-Tyr have been determined simultaneously in a real human autopsy sample from a victim of chlorine exposure.

DNA hydroxymethylation reprogramming of β-oxidation genes mediates early-life arsenic-evoked hepatic lipid accumulation in adult mice

The metabolic disorders are becoming an epidemic disease endangering public health in countries. Environmental factors are mainly reason for the growth of metabolic disorders. Previous research suggests that DNA methylation is a potential mechanism. Recently, it has been reported that DNA hydroxymethylation is also a stable marker of epigenetic reprogramming. Hence, the study aims to investigate whether DNA hydroxymehylation mediates early-life environmental stress-evoked metabolic disorder in adulthood. Mice were orally administered with arsenic (As), an environmental stressor, throughout pregnancy. We show that early-life As exposure induces glucose intolerance and hepatic lipid accumulation in adulthood. Early-life As exposure alters epigenetic reprogramming and expression of lipid metabolism-related genes including β-oxidation-specific genes in adulthood. Of interest, early-life As exposure alters epigenetic reprogramming of hepatic lipid metabolism partially through reducing DNA hydroxymethylation modification of β-oxidation-related genes in developing liver. Mechanistically, early-life As exposure suppresses ten-eleven translocation (TET) activity through downregulating isocitrate dehydrogenases (Idh) and reducing alpha-ketoglutarate (α-KG) content in the developing liver. In addition, early-life As exposure inhibits TET1 binding to CpG-rich fragments of β-oxidation-related genes in developing liver. This study provide novel evidence that early-life environmental stress leads to later life metabolic disorders by altering hepatic DNA hydroxymethylation reprogramming.

Circulating microRNA miR-137 as a stable biomarker for methamphetamine abstinence

ObjectiveStimulant use instigates abstinence syndrome in humans. miRNAs are a critical component for the pathophysiology of stimulant abstinence. Here we sought to identify a miRNA marker of methamphetamine abstinence in the circulating extracellular vesicles (cEVs).MethodsmiR-137 in the cEVs was quantified by qPCR in thirty-seven patients under methamphetamine abstinence and thirty-five age-matched healthy controls recruited from 2014 to 2016 from the general adult population in a hospital setting, Seoul, South Korea. Diagnostic power was evaluated by area under curve in the receiver-operating characteristics curve and other multiple statistical parameters.ResultsPatients under methamphetamine abstinence exhibited a significant reduction in cEV miR-137. Overall, cEV miR-137 had high potential as a blood-based marker of methamphetamine abstinence. cEV miR-137 retained the diagnostic power irrespective of the duration of methamphetamine abstinence or methamphetamine use. Interestingly, cEV miR-137 interacted with age: Control participants displayed an aging-dependent reduction of cEV miR-137, while methamphetamine-abstinent patients showed an aging-dependent increase in cEV miR-137. Accordingly, cEV miR-137 had variable diagnostic power depending on age, in which cEV miR-137 more effectively discriminated methamphetamine abstinence in the younger population. Duration of methamphetamine use or abstinence, cigarette smoking status, depressive disorder, or antidepressant treatment did not interact with the methamphetamine abstinence-induced reduction of cEV miR-137.ConclusionOur data collectively demonstrated that miR-137 in the circulating extracellular vesicles held high potential as a stable and accurate diagnostic marker of methamphetamine abstinence syndrome.

Abstract TP192: Plasma Mid-regional Pro-adrenomedullin Reflects Ischemic Penumbra In Acute Ischemic Stroke

Background: Adrenomedullin (AM), an endogenous peptide, is secreted in response to cerebral ischemia, contributing to neuroprotection by exerting anti-inflammatory and vasodilatory effects. Although previous studies showed elevated levels of mid-regional fragment of pro-AM (MR-proAM), a stable marker of AM, predict unfavorable outcome in acute ischemic stroke (AIS), the role of the biomarker remains to be determined. Methods: Firstly, plasma MR-proAM levels were measured in consecutively enrolled patients with AIS within 4.5 hours of the onset and compared with those of healthy controls. Propensity score was used to match the baseline difference. Secondly, variables associated with the increased MR-proAM levels were evaluated in AIS. Finally, it was determined whether MR-proAM levels are associated with a large penumbra which was defined as ICA/M1 occlusion with NIH stroke scale ≥ 6 and ischemic core volume ≤ 50 ml calculated by the RAPID software. Results: MR-proAM levels in AIS (n = 122; median age, 77 years; male, 59%; median NIHSS 6.5) were higher (median 0.68 vs. 0.42 nmol/L, p < 0.001) compared with controls (n = 1298; median age, 58 years; male, 33%). The difference remained after matching of baseline (p=0.004). In patients with AIS, MR-proAM levels were correlated with NIHSS (r = 0.367, p < 0.001) and eGFR (r = -0.527, p < 0.001). MR-proAM levels did not correlate with ischemic core volume (r = 0.065, p = 0.478), but significantly increased in AIS patients with cardioembolic stroke (CES) (n = 53) (0.80 vs. 0.59 nmol/L, p < 0.001) and ICA/M1 occlusion (n = 40) (0.78 vs. 0.63 nmol/L, p = 0.048). Interestingly, MR-proAM levels were higher (0.85 vs. 0.62, p < 0.001) in AIS patients with a large penumbra (n = 24) compared to those without (n = 98), and the difference remained after adjustment of baseline factors (OR [95% confidence interval], 1.43 [1.10-2.02], p = 0.004). Conclusions: MR-proAM levels increase in patients with AIS, especially in those with large vessel occlusion and clinical imaging mismatch. MR-proAM is a potential biomarker for ischemic penumbra.

Genome-wide association mapping of genotype-environment interactions affecting yield-related traits of spring wheat grown in three watering regimes

Genotype-environment interaction (GxE) has a great impact on wheat physiology, morphology and grain yield (GY). We evaluated an association mapping panel of spring wheat advanced lines for chlorophyll content, canopy temperature (CT), and yield-related traits under three different watering regimes in two consecutive growing seasons. Genome-wide association mapping identified 457 SNPs, with significant effects that varied with the watering regimes and growing seasons, of which 199 and 69 SNPs showed pleiotropic and conditionally neutral effects, respectively, on the measured traits. We mapped 61 SNPs with effects higher than 10% on all traits, showing antagonistic pleiotropic effects on CT, corresponding to 46 genes; some of these genes represent good candidates to control wheat response to water availability. Surprisingly, no significant SNPs were mapped in the semi-dwarfing genes, Rht-B1b or Rht-D1b . However, haplotype analysis of the SNPs located at the positions of both genes revealed significant interactions of GY with the watering regimes for Rht-B1b and with the growing season for Rht-D1b . We selected genotypes that outperformed two local check cultivars; some of them overlapped across the three watering regimes and could be used to create a multi-parent population to further unravel the genetic factors underlying yield component traits across drought stress. Our results demonstrate the importance of incorporating GxE in mapping models to better understand wheat response to different watering regimes and to select stable markers for selection. • Three genotypes outperformed two local check cultivars under all treatments, whereas the remaining genotypes showed GxE. • Out of the 457 significant SNPs, 199 and 69 SNPs showed pleiotropic and conditionally neutral effects, respectively. • 61 main-effect SNPs can assist significantly in marker-assisted breeding programs. • Two favorable QTLs on 5D and 7D increased GY, and as an adaptive strategy to cope with drought, lowered CT.

Urinary metabolic biomarkers of diet quality in European children are associated with metabolic health.

Urinary metabolic profiling is a promising powerful tool to reflect dietary intake and can help understand metabolic alterations in response to diet quality. Here, we used 1H NMR spectroscopy in a multicountry study in European children (1147 children from 6 different cohorts) and identified a common panel of 4 urinary metabolites (hippurate, N-methylnicotinic acid, urea, and sucrose) that was predictive of Mediterranean diet adherence (KIDMED) and ultra-processed food consumption and also had higher capacity in discriminating children's diet quality than that of established sociodemographic determinants. Further, we showed that the identified metabolite panel also reflected the associations of these diet quality indicators with C-peptide, a stable and accurate marker of insulin resistance and future risk of metabolic disease. This methodology enables objective assessment of dietary patterns in European child populations, complementary to traditional questionary methods, and can be used in future studies to evaluate diet quality. Moreover, this knowledge can provide mechanistic evidence of common biological pathways that characterize healthy and unhealthy dietary patterns, and diet-related molecular alterations that could associate to metabolic disease.

The application of the skin virome for human identification

The use of skin virome offers a unique approach for human identification purposes in instances where a viable and statistically relevant human DNA profile is unavailable. The skin virome may act as an alternative DNA profile and/or an additional form of probative genetic material. To date, no study has attempted to investigate the human virome over a time series across various physical locations of the body to identify its diagnostic potential as a tool for human identification. For this study, we set out to evaluate the stability, diversity, and individualization of the human skin virome. An additional goal was to identify putative viral signatures that can be used in conjunction with traditional forensic STR loci. In order to accomplish this, human viral metagenomes were collected and sequenced from 42 individuals at three anatomical locations (left hand, right hand, and scalp) across multiple collection periods over a 6-month window of time. Assembly dependent and independent bioinformatic approaches, along with a database centered assessment of viral identification, resulted in three sets of stable putative viral markers. In total, with the three sets combined, we identified 59 viral biomarker regions, consisting of viral species and uncharacterized viral genome assemblies, that were stable over the sampling period. Additionally, we found the abundance profiles of these 59 viral biomarkers, based on presence or absence, to be significantly different across subjects (P < 0.001). Here we demonstrate that not only is the human virome applicable to be used for human identification, but we have identified many viral signatures that can putatively be used for forensic applications, thus providing a foundation to the novel field of forensic virology.

Loss of 5-hydroxymethylcytosine as a Poor Prognostic Factor for Primary Testicular Diffuse Large B-cell Lymphoma.

Background: 5-Hydroxymethylcytosine (5-hmC), a stable epigenetic marker, is closely related to tumor staging, recurrence and survival, but the prognostic value of 5-hmC in primary testicular diffuse large B-cell lymphoma (PT-DLBCL) remains unclear. This study aimed to investigate the 5-hmC expression in PT-DLBCL and evaluate its prognostic value.Methods: A total of 34 patients with PT-DLBCL treated in the Department of Hematology from August 2000 to August 2020 were included in this study. The expression of 5-hmC in PT-DLBCL tissues and normal testicular tissues were assessed by immunohistochemistry. 5-hmC staining is estimated as a percentage under every nuclear staining intensity score (0-3), 0 or 1 of which were regarded as 5-hmC reduction. The quantification of 5-hmC reduction is defined as the percentage of cells with 5-hmC staining scores of 0 and 1. According 5-hmC reduction of 80%, a 5-hmC reduction of <80% is regarded as "5-hmC high-level group", and a 5-hmC reduction of ≥80% is regarded as "5-hmC low-level group". Furthermore, Cox regression model was used to evaluate the prognostic value of all covariates.Results: The median percentage of 5-hmC reduction in the PT-DLBCL group was 77.50% (60%-90%), the median 5-hmC reduction in the normal testicular tissues was 30% (20%-50%). Compared with normal testicular tissue, 5-hmC levels in PT-DLBCL tissue were significantly decreased (p<0.05). Of the 34 PT-DLBCL patients, 17 had tumors with relatively low 5-hmC expression (5-hmC reduction of ≥80%) and 17 had tumors with relatively high 5-hmC expression (5-hmC reduction of < 80%). 5-hmC expression was negatively correlated with international prognostic index (p = 0.037), while there was no significant difference in 5-hmC decrease among different groups of age at diagnosis, lactate dehydrogenase, testicular lymphoma involvement (unilateral or bilateral), Ki-67 and tumor diameter. Relatively low 5-hmC expression indicated shorter overall survival (OS) (5-year OS 50.2% vs 81.3%, p=0.022) and progression-free survival (PFS) (5-year PFS 38.5% vs 70.7%, p=0.001). Cox multivariate analysis of IPI (2-3 vs. 0-1), intrathecal prophylaxis (No vs. Yes), and 5-hmC reduction (≥80% vs. <80%) showed that 5-hmC reduction ≥80% (hazard ratio: 7.252, p = 0.005) and not receiving intrathecal prophylaxis (hazard ratio: 7.207, p =0.001) are independent risk factors for poor prognosis of PT-DLBCL.Conclusion: Our results suggested that 5-hmC decline can be identified as a poor prognostic predictor for PT-DLBCL. It is necessary to further explore the underlying mechanism of this epigenetic marker to identify methods to re-establish 5-hmC levels and provide new targets for cancer therapy.

Bibliometric analysis of isotopic studies on Quaternary megafauna available in the Scopus database.

The number of isotopic studies on Quaternary megafauna has increased over the last decades, yet, there is no published data addressing the status of scientific production of this research field. The present study shows the results of a bibliometric research carried out in the Scopus database where the publishing trends within this scientific field was analyzed using the open source software tool SciMAT. We retrieved 278 papers published from 1980 to 2019 and observed that a significant increase in publishing has mainly occurred in the last decade analyzed here. We also identified some of the field´s most influential articles and journals; recognized that carbon, oxygen and nitrogen isotopes are the most used markers in these studies; and that the most cited taxa are representatives of Equidae, Bovidae and Proboscidae. Also, Paleoecology is the basic thematic area, whereas Climate and Paleoenvironmental Changes is the one with the greatest development potential. Our results clearly show that the isotopic study on Quaternary megafauna is still under development and that some subjects could be further explored, such as analyzing more taxa within Carnivora, Pilosa, Notoungulata, Cetartiodactyla and Perissodactyla as well as using other less frequent stable isotope markers, such as strontium, calcium and hydrogen.

Using molecular and stable isotope markers to identify the main predators of Nephrops norvegicus in Mediterranean deep-water ecosystems

To obtain a better understanding of the functioning of ecosystems and how they respond to disturbance, it is necessary to identify the relevant biotic interactions and specific trophic roles. Predation is one of the most important biotic interactions that can also define the spatial patterns of other species. Many predators are considered key species for the functioning and maintenance of ecosystems, as they play an important ecological role that can influence the dynamics at lower trophic levels. The Norway lobster Nephrops norvegicus is one of the most valued European fishing stocks. However, its value and capture have declined over the last decade. In the Atlantic Ocean, Atlantic cod Gadus morhua is the main predator of N. norvegicus. However, this species is not present in the Mediterranean Sea, and little is known about which species might prey on N. norvegicus in this area. Here, we combine 2 methodologies—genetic identification of stomach contents and stable isotope analyses—to identify, for the first time, the main predators of N. norvegicus in the Mediterranean Sea. Moreover, we have created the Predation Index, which determines the most influential predator affecting N. norvegicus population dynamics. Our results reveal that the major predators are the cephalopods Sepietta spp. and Abralia veranyi, which probably affect the early stages of N. norvegicus, followed by the elasmobranch Scyliorhinus canicula and the bony fishes Merluccius merluccius, Trigla lyra, and Conger conger. To evaluate possible fluctuations in the N. norvegicus population, we consider the assessment of these predator populations crucial.

Analytic consistency and neural correlates of peak alpha frequency in the study of pain

BackgroundSeveral studies have found evidence of reduced resting-state peak alpha frequency (PAF) in populations with pain. However, the stability of PAF from different analytic pipelines used to study pain has not been determined and underlying neural correlates of PAF have not been validated in humans. New methodFor the first time we compare analytic pipelines and the relationship of PAF to activity in the whole brain and thalamus, a hypothesized generator of PAF. We collected resting-state functional magnetic resonance imaging (rs-fMRI) data and subsequently 64 channel resting-state electroencephalographic (EEG) from 47 healthy men, controls from an ongoing study of chronic prostatitis (a pain condition affecting men). We identified important variations in EEG processing for PAF from a review of 17 papers investigating the relationship between pain and PAF. We tested three progressively complex pre-processing pipelines and varied four postprocessing variables (epoch length, alpha band, calculation method, and region-of-interest [ROI]) that were inconsistent across the literature. ResultsWe found a single principal component, well-represented by the average PAF across all electrodes (grand-average PAF), explained > 95% of the variance across participants. We also found the grand-average PAF was highly correlated among the pre-processing pipelines and primarily impacted by calculation method and ROI. Across methods, interindividual differences in PAF were correlated with rs-fMRI-estimated activity in the thalamus, insula, cingulate, and sensory cortices. ConclusionsThese results suggest PAF is a relatively stable marker with respect to common pre and post-processing methods used in pain research and reflects interindividual differences in thalamic and salience network function.

Identification of Candidate Genes and Genomic Regions Associated with Adult Plant Resistance to Stripe Rust in Spring Wheat

Wheat stripe rust (caused by Puccinia striiformis f. sp. tritici) is a major disease that damages wheat plants and affects wheat yield all over the world. In recent years, stripe rust became a major problem that affects wheat yield in Egypt. New races appeared and caused breakdowns in the resistant genotypes. To improve resistance in the Egyptian genotypes, new sources of resistance are urgently needed. In the recent research, a set of 95 wheat genotypes collected from 19 countries, including Egypt, were evaluated for their resistance against the Egyptian race(s) of stripe rust under field conditions in the two growing seasons 2018/2019 and 2019/2020. A high genetic variation was found among the tested genotypes. Single marker analysis was conducted using a subset of 71 genotypes and 424 diversity array technology (DArT) markers, well distributed across the genome. Out of the tested markers, 13 stable markers were identified that were significantly associated with resistance in both years (p-value ≤ 0.05). By using the sequence of the DArT markers, the chromosomal position of the significant DArT markers was detected, and nearby gene models were identified. Two markers on chromosomes 5A and 5B were found to be located within gene models functionally annotated with disease resistance in plants. These two markers could be used in marker-assisted selection for stripe rust resistance under Egyptian conditions. Two German genotypes were carrying the targeted allele of all the significant DArT markers associated with stripe rust resistance and could be used to improve resistance under Egyptian conditions.

Systematic Analysis of an Invasion-Related 3-Gene Signature and Its Validation as a Prognostic Model for Pancreatic Cancer.

BackgroundPancreatic adenocarcinoma (PAAD) is a malignant tumor of the digestive system that is associated with a poor prognosis in patients owing to its rapid progression and high invasiveness.MethodsNinety-seven invasive-related genes obtained from the CancerSEA database were clustered to obtain the molecular subtype of pancreatic cancer based on the RNA-sequencing (RNA-seq) data of The Cancer Genome Atlas (TCGA). The differentially expressed genes (DEGs) between subtypes were obtained using the limma package in R, and the multi-gene risk model based on DEGs was constructed by Lasso regression analysis. Independent datasets GSE57495 and GSE62452 were used to validate the prognostic value of the risk model. To further explore the expression of the hub genes, immunohistochemistry was performed on PAAD tissues obtained from a large cohort.ResultsThe TCGA-PAAD samples were divided into two subtypes based on the expression of the invasion-related genes: C1 and C2. Most genes were overexpressed in the C1 subtype. The C1 subtype was mainly enriched in tumor-related signaling pathways, and the prognosis of patients with the C1 subtype was significantly worse than those with the C2 subtype. A 3-gene signature consisting of LY6D, BCAT1, and ITGB6 based on 538 DEGs between both subtypes serves as a stable prognostic marker in patients with pancreatic cancer across multiple cohorts. LY6D, BCAT1, and ITGB6 were over-expressed in 120 PAAD samples compared to normal samples.ConclusionsThe constructed 3-gene signature can be used as a molecular marker to assess the prognostic risk in patients with PAAD.

Aminoferrocene‐Based Anticancer Prodrugs Labelled with Cyanine Dyes for in vivo Imaging

AbstractN‐alkylaminoferrocene‐based (NAAF) prodrugs are activated in the presence of reactive oxygen species (ROS), based on which these prodrugs target cancer cells (high ROS) and do not affect normal cells (low ROS). To gain some insights into their mode of action in vivo, we have investigated the biodistribution of 18F‐labelled NAAF prodrugs in tumor‐bearing mice by positron emission tomography (PET). Due to the short half‐life of 18F, the experimental time frame was restricted to 60 min. To extend the observation window, a more stable marker is required. In this paper, we report on conjugates of NAAF prodrugs with two cyanine dyes Cy5 and Cy7 including details of their synthesis, characterization and basic properties in cell free settings and their cellular uptake in representative human cancer cells. Finally, the Cy5 conjugate was subjected to in vivo fluorescence imaging studies to determine the prodrug biodistribution over 24 h.

Transcriptome sequencing analysis for the identification of stable lncRNAs associated with bovine Staphylococcus aureus mastitis

BackgroundStaphylococcus aureus (S. aureus) mastitis is one of the most difficult diseases to treat in lactating dairy cows worldwide. S. aureus with different lineages leads to different host immune responses. Long non-coding RNAs (lncRNAs) are reported to be widely involved in the progress of inflammation. However, no research has identified stable lncRNAs among different S. aureus strain infections. In addition, folic acid (FA) can effectively reduce inflammation, and whether the inflammatory response caused by S. aureus can be reduced by FA remains to be explored.MethodslncRNA transcripts were identified from Holstein mammary gland tissues infected with different concentrations of S. aureus (in vivo) and mammary alveolar cells (Mac-T cells, in vitro) challenged with different S. aureus strains. Differentially expressed (DE) lncRNAs were evaluated, and stable DE lncRNAs were identified in vivo and in vitro. On the basis of the gene sequence conservation and function conservation across species, key lncRNAs with the function of potentially immune regulation were retained for further analysis. The function of FA on inflammation induced by S. aureus challenge was also investigated. Then, the association analysis between these keys lncRNA transcripts and hematological parameters (HPs) was carried out. Lastly, the knockdown and overexpression of the important lncRNA were performed to validate the gene function on the regulation of cell immune response.ResultsLinear regression analysis showed a significant correlation between the expression levels of lncRNA shared by mammary tissue and Mac-T cells (P < 0.001, R2 = 0.3517). lncRNAs PRANCR and TNK2–AS1 could be regarded as stable markers associated with bovine S. aureus mastitis. Several HPs could be influenced by SNPs around lncRNAs PRANCR and TNK2–AS1. The results of gene function validation showed PRANCR regulates the mRNA expression of SELPLG and ITGB2 within the S. aureus infection pathway and the Mac-T cells apoptosis. In addition, FA regulated the expression change of DE lncRNA involved in toxin metabolism and inflammation to fight against S. aureus infection.ConclusionsThe remarkable association between SNPs around these two lncRNAs and partial HP indicates the potentially important role of PRANCR and TNK2–AS1 in immune regulation. Stable DE lncRNAs PRANCR and TNK2–AS1 can be regarded as potential targets for the prevention of bovine S. aureus mastitis. FA supplementation can reduce the negative effect of S. aureus challenge by regulating the expression of lncRNAs.

Quantitative assessment of contrast-enhancement patterns of the healthy dental pulp by magnetic resonance imaging: A prospective in vivo study.

This prospective in vivo study aimed to optimize the assessment of pulpal contrast-enhancement (PCE) on dental magnetic resonance imaging (dMRI) and investigate physiological PCE patterns. In 70 study participants, 1585 healthy teeth were examined using 3-Tesla dMRI before and after contrast agent administration. For all teeth, the quotient of post- and pre-contrast pulp signal intensity (Q-PSI) was calculated to quantify PCE. First, pulp chambers were analysed in 10 participants to compare the coefficient of variation of mean versus maximum Q-PSI values (Q-PSImean versus Q-PSImax ). Second, dynamic PCE was evaluated in 10 subjects to optimize the time interval between contrast agent application and image acquisition. Finally, 50 participants (age groups: 20-29, 30-39, 40-49, 50-59 and 60-69years) were examined to analyse age, gender, tooth types and maxilla versus mandible as independent factors of PCE. Statistical analysis was performed using Wilcoxon signed rank test and linear mixed models. PCE assessment based on Q-PSImax was associated with a significantly smaller coefficient of variation compared with Q-PSImean , with median values of 0.17 versus 0.21 (p=.002). Analysis of dynamic PCE revealed an optimal timing interval for image acquisition 4min after contrast media application. No significant differences in PCE were observed by comparing age groups, female versus male participants and maxillary versus mandibular teeth (p>.05). Differences between tooth types were small (median Q-PSImax values of 2.52/2.32/2.30/2.20 for molars/premolars/canines/incisors) but significant (p<.05), except for the comparison of canines versus premolars (p=.80). PCE in dMRI was a stable intra-individual marker with only minor differences between different tooth types, thus forming an important basis for intra-individual controls when assessing teeth with suspected endodontic pathosis. Furthermore, it was demonstrated that PCE is independent of age, gender and jaw type. These findings indicate that dMRI-based PCE analysis could be a valuable diagnostic tool for the identification of various pulp diseases in future patient studies.

Changes in Factors Regulating Serum Sodium Homeostasis During Two Ultra-Endurance Mountain Races of Different Distances: 69km vs. 121km.

Overdrinking and non-osmotic arginine vasopressin release are the main risk factors for exercise-associated hyponatremia (EAH) in ultra-marathon events. However, particularly during ultra-marathon running in mountainous regions, eccentric exercise and hypoxia, which have been shown to modulate inflammation, hormones regulating fluid homeostasis (hypoxia), and oxidative stress, could contribute to serum sodium changes in a dose-dependent manner. To the best of our knowledge, the contribution of these factors, the extent of which depends on the duration and geographical location of the race, has not been well studied. Twelve male participants (11 finishers) of the short (69km, 4,260m elevation-gain) and 15 male participants (seven finishers) of the long (121km, 7,554m elevation-gain) single-stage Südtirol Ultra Sky-Race took part in this observational field study. Venous blood was drawn immediately before and after the race. Analyses included serum sodium concentration, copeptin (a stable marker for vasopressin), markers of inflammation, muscle damage and oxidative stress. Heart rate was measured during the race and race time was obtained from the race office. During the short and the long competition two and one finishers, respectively showed serum sodium concentrations >145mmol/L. During the long competition, one athlete showed serum sodium concentrations <135mmol/L. Only during the short competition percent changes in serum sodium concentrations of the finishers were related to percent changes in body mass (r=−0.812, p=0.002), total time (r=−0.608, p=0.047) and training impulse (TRIMP) (r=−0.653, p=0.030). Data show a curvilinear (quadratic) relationship between percent changes in serum sodium concentration and body mass with race time when including all runners (short, long, finishers and non-finishers). The observed prevalence of hypo- and hypernatremia is comparable to literature reports, as is the relationship between serum sodium changes and race time, race intensity and body mass changes of the finishers of the short race. The curvilinear relationship indicates that there might be a turning point of changes in serum sodium and body mass changes after a race time of approximately 20h. Since the turning point is represented mainly by non-finishers, regardless of race duration slight decrease in body mass and a slight increase in serum sodium concentration should be targeted to complete the race. Drinking to the dictate of thirst seems an adequate approach to achieve this goal.

Existence of Functional Connectome Fingerprint during Infancy and Its Stability over Months.

The functional connectome fingerprint is a cluster of individualized brain functional connectivity patterns that are capable of distinguishing one individual from others. Although its existence has been demonstrated in adolescents and adults, whether such individualized patterns exist during infancy is barely investigated despite its importance in identifying the origin of the intrinsic connectome patterns that potentially mirror distinct behavioral phenotypes. To fill this knowledge gap, capitalizing on a longitudinal high-resolution structural and resting-state functional MRI dataset with 104 human infants (53 females) with 806 longitudinal scans (age, 16-876 d) and infant-specific functional parcellation maps, we observe that the brain functional connectome fingerprint may exist since infancy and keeps stable over months during early brain development. Specifically, we achieve an ∼78% individual identification rate by using ∼5% selected functional connections, compared with the best identification rate of 60% without connection selection. The frontoparietal networks recognized as the most contributive networks in adult functional connectome fingerprinting retain their superiority in infants despite being widely acknowledged as rapidly developing systems during childhood. The existence and stability of the functional connectome fingerprint are further validated on adjacent age groups. Moreover, we show that the infant frontoparietal networks can reach similar accuracy in predicting individual early learning composite scores as the whole-brain connectome, again resembling the observations in adults and highlighting the relevance of functional connectome fingerprint to cognitive performance. For the first time, these results suggest that each individual may retain a unique and stable marker of functional connectome during early brain development.SIGNIFICANCE STATEMENT Functional connectome fingerprinting during infancy featuring rapid brain development remains almost uninvestigated even though it is essential for understanding the early individual-level intrinsic pattern of functional organization and its relationship with distinct behavioral phenotypes. With an infant-tailored functional connection selection and validation strategy, we strive to provide the delineation of the infant functional connectome fingerprint by examining its existence, stability, and relationship with early cognitive performance. We observe that the brain functional connectome fingerprint may exist since early infancy and remains stable over months during the first 2 years. The identified key contributive functional connections and networks for fingerprinting are also verified to be highly predictive for cognitive score prediction, which reveals the association between infant connectome fingerprint and cognitive performance.

Selecting Processing Robust Markers Using High-Resolution Mass Spectrometry for the Detection of Milk in Food Products.

Cow's milk allergy is one of the most reported food allergies in Europe. To help patients suffering from food allergies it is important to be able to detect milk in different foods. An analytical method that is gaining interest in the field of allergen detection is ultrahigh performance liquid chromatography-tandem mass spectrometry, where the analyte is a target peptide. When these peptide biomarkers are selected, the effect of food processing should be taken into account to allow a robust detection method. This work aims at identifying such processing stable peptide markers for milk for the ultrahigh performance liquid chromatography-tandem mass spectrometry based detection of food allergens in different food products. Milk-incurred food materials that underwent several processing techniques were produced. This was followed by establishing tryptic peptide profiles from each matrix using ultrahigh performance liquid chromatography-high resolution mass spectrometry. A careful comparison of peptide profiles/intensities and the use of specific exclusion criteria resulted in the selection of eight peptide biomarkers suitable for application in ultrahigh performance liquid chromatography-tandem mass spectrometry based milk detection methods. One of these markers is an α-lactalbumin specific peptide, which has been determined to be stable in different incurred materials for the first time. To our knowledge, this is the first systematic and experimentally based approach for the selection of suitable milk peptide biomarkers robust toward multiple, often applied food processing techniques for milk. Ensuring the exact knowledge of the food processing circumstances by starting from well-defined raw material and using fully controlled settings to produce incurred test material allowed the construction of a peptide database with robust markers. These robust markers can be used for the development of a robust detection method for milk in different food matrixes. To facilitate food allergen detection in processed food, processing stable peptide markers for the detection of milk in food products were determined using Ultra-High Performance Liquid Chromatography-High Resolution Mass Spectrometry on well-defined raw materials which were processed in accordance with often used processing techniques.