Mosaic loss of chromosome Y (LOY) is the most common somatic genetic aberration and is associated with increased risk for all-cause mortality, various forms of cancer and Alzheimer's disease, as well as other common human diseases. By tracking LOY frequencies in subjects from which blood samples have been serially collected up to five times during up to 22 years, we observed a pronounced intra-individual variation of changes in the frequency of LOY within individual men over time. We observed that in some individuals the frequency of LOY in blood clearly progressed over time and that in other men, the frequency was constant or showed other types of longitudinal development. The predominant method used for estimating LOY is calculation of the median Log R Ratio of probes located in the male specific part of chromosome Y (mLRRY) from intensity data generated by SNP-arrays, which is difficult to interpret due to its logarithmic and inversed scale. We present here a formula to transform mLRRY-values to percentage of LOY that is a more comprehensible unit. The formula was derived using measurements of LOY from matched samples analysed using SNP-array, whole genome sequencing and a new AMELX/AMELY-based assay for droplet digital PCR. The methods described could be applied for analyses of the vast amount of SNP-array data already generated in the scientific community, allowing further discoveries of LOY associated diseases and outcomes.

With the recognition that most global environmental problems are a result of human actions, there is an increasing interest in approaches which have the potential to influence human behaviour. Images have a powerful role in shaping persuasive messages, yet research on the impacts of visual representations of nature is a neglected area in biodiversity conservation. We systematically screened existing studies on the use of animal imagery in conservation, identifying 37 articles. Although there is clear evidence that images of animals can have positive effects on people's attitudes to animals, overall there is currently a dearth of accessible and comparable published data demonstrating the efficacy of animal imagery. Most existing studies are place and context-specific, limiting the generalisable conclusions that can be drawn. Transdisciplinary research is needed to develop a robust understanding of the contextual and cultural factors that affect how animal images can be used effectively for conservation purposes.

Influence of an intracranial aneurysm (IA) rupture on the expression of miRNAs and the potential significance of the resulting changes remains poorly understood. We aimed to characterize the response to the IA rupture through the analysis of miRNAs in peripheral blood cells. Expression of small RNAs was investigated using deep transcriptome sequencing in patients in the acute phase of an IA rupture (first 72 h), in the chronic phase (3-15 months), and controls. A functional analysis and the potential interactions between miRNAs and target genes were investigated. We also measured the levels of proteins that were influenced by regulated miRNAs. We found that 106 mature miRNAs and 90 miRNA precursors were differentially expressed among the groups. The regulated miRNAs were involved in a variety of pathways, and the top pathway involved cytokine-cytokine receptor interactions. The identified miRNAs targeted the inflammatory factors HMGB1 and FASLG. Changes in their expression were detected at the mRNA and protein levels. IA rupture strongly influences the transcription profiles in peripheral blood cells. The regulated miRNAs were involved in the control of immune cell homeostasis. In summary, these results may aid in the elucidation of the molecular mechanisms that orchestrate the inflammatory response to IA rupture.

The purpose of this study was to determine the long axes of molars with multiple roots through ordinary least squares regression (LSR) and to compare them with the axes defined by principal component analysis (PCA). Three-dimensional radiological images of 20 dry skulls were obtained by cone-beam computed tomography (CBCT). Data from maxillary and mandibular first molars were extracted from the CBCT DICOM data with a three-dimensional image visualization system. The obtained data were reconstructed, converted to STL files, and three-dimensional coordinate values were extracted. The long axes were estimated by an algorithm to synchronize the LSR line with the horizontal axis which was translated to the vertical axis. The axes of the molars defined by LSR were compared with the axes of the molars defined by PCA. The coordinate point number of each molar was 5400-5800. The algorithm for determining the tooth axes in this study consisted of four stages containing three steps each. The distance between the two axes calculated by the two methods (LSR and PCA) on the horizontal plane through the origin was less than 10-12 mm and the deviations between them were less than 0.003°. The long axes of the molars estimated by LSR agree almost exactly with the axes estimated by PCA, and the accuracy is sufficient for clinical usage; however, the distance between them would shorten with a more severe convergence condition of the α value at each stage of this LSR system.

Bruton's tyrosine kinase (BTK) is a key regulator of B cell receptor and Fc receptor signaling, and a rational therapeutic target for autoimmune diseases. This first-in-human phase I, double-blind, placebo-controlled trial investigated the safety, tolerability, pharmacokinetics (PK), target occupancy, and effects on QT interval of evobrutinib, a highly selective, oral inhibitor of BTK, in healthy subjects. This dose escalation trial consisted of two parts. Part 1 included 48 subjects in 6 ascending dose cohorts (25, 50, 100, 200, 350, and 500 mg) randomized to a single dose of evobrutinib or placebo. Part 2 included 36 subjects in 3 ascending dose cohorts (25, 75, and 200 mg/day) randomized to evobrutinib or placebo once daily for 14 days. Safety and tolerability, as well as PK and target occupancy (total and free BTK in peripheral blood mononuclear cells), were assessed following single and multiple dosing. PK parameters were determined by noncompartmental methods. QT interval was obtained from 12-lead electrocardiogram recordings and corrected for heart rate by Fridericia's method (QTcF). Treatment-emergent adverse events (TEAEs) were mostly mild, occurring in 25% of subjects after single dosing, and 48.1% after multiple dosing. There was no apparent dose relationship regarding frequency or type of TEAE among evobrutinib-treated subjects. Absorption was rapid (time to reach maximum plasma concentration (Tmax ) ~ 0.5 hour), half-life short (~ 2 hours), and PK dose-proportional, with no accumulation or time dependency on repeat dosing. BTK occupancy was dose-dependent, reaching maximum occupancy of > 90% within ~ 4 hours after single doses ≥ 200 mg; the effect was long-lasting (> 50% occupancy at 96 hours with ≥ 100 mg). After multiple dosing, full BTK occupancy was achieved with 25 mg, indicating slow turnover of BTK protein in vivo. Concentration-QTcF analyses did not show any impact of evobrutinib concentration on corrected QT (QTc). In summary, evobrutinib was well-tolerated, showed linear and time-independent PK, induced long-lasting BTK inhibition, and was associated with no prolongation of QT/QTc interval in healthy subjects. Evobrutinib is, therefore, suitable for investigation in autoimmune diseases.

We aim to characterize the kinetics of early and late microglial phenotypes after systemic inflammation in an animal model of severe sepsis and the effects of minocycline on these phenotypes. Rats were subjected to CLP, and some animals were treated with minocycline (10 ug/kg) by i.c.v. administration. Animals were killed 24 hours, 5, 10 and 30 days after sepsis induction, and serum and hippocampus were collected for subsequent analyses. Real-time PCR was performed for M1 and M2 markers. TNF-α, IL-1β, IL-6, IL-10, CCL-22 and nitrite/nitrate levels were measured. Immunofluorescence for IBA-1, CD11b and arginase was also performed. We demonstrated that early after sepsis, there was a preponderant up-regulation of M1 markers, and this was not switched to M2 phenotype markers later on. We found that up-regulation of both M1 and M2 markers co-existed up to 30 days after sepsis induction. In addition, minocycline induced a down-regulation, predominantly, of M1 markers. Our results suggest early activation of M1 microglia that is followed by an overlap of both M1 and M2 phenotypes and that the beneficial effects of minocycline on sepsis-associated brain dysfunction may be related to its effects predominantly on the M1 phenotype.

The peripheral nervous system is involved in a multitude of physiological functions. Recording neural signals provides information that can be used by diagnostic bioelectronic medicine devices, closed-loop neuromodulation therapies and other neuroprosthetic applications. The ability to accurately record these signals is challenging, due to the presence of various biological and instrument-related interference sources.

Developments in proximity ligation methods and sequencing technologies have provided high-resolution views of the organization of the genome inside the nucleus. A prominent feature of Hi-C maps is regions of increased self-interaction called topologically associating domains (TADs). Despite the strong evolutionary conservation and clear link with gene expression, the exact role of TADs and even their definition remains debatable. Here, I review the discovery of TADs, how they are commonly identified, and the mechanisms that lead to their formation. Furthermore, I discuss recent results that have created a more nuanced view of the role of TADs in the regulation of genes. In light of this, I propose that when we define TADs, we also consider the mechanisms that shape them.

There is currently no agreement on which factor modulates most effectively and enduringly brain plasticity in bilingual individuals. Grouping heterogeneous linguistic profiles under a dichotomous condition (bilingualism versus monolingualism) may obscure critical aspects of language experience underlying neural changes, thus leading to variable and often conflicting findings. In the present study, we overcome these limitations by analyzing the individual and joint contribution of L2 AoA, proficiency and usage - all measured as continuous variables - on the resting-state functional connectivity of the brain networks mediating the specific demands of bilingual language processing: the language network and the executive control network. Our results indicate that bilingual experience - defined as a continuous and multifaceted phenomenon - impacts brain plasticity by modulating the functional connectivity both within and between language and control networks. Each experience-related factor considered played a role in changing the connectivity of these regions. Moreover, the effect of AoA was modulated by proficiency and usage. These findings shed new light on the importance of modeling bilingualism as a gradient measure rather than an all-or-none phenomenon.

Activation of renin-angiotensin- system, nitric oxide (NO•) bioavailability and subsequent sympathoexcitation plays a pivotal role in the pathogenesis of many cardiovascular diseases, including hypertension. Previously we have shown increased protein expression of PIN (a protein inhibitor of nNOS: neuronal nitric oxide synthase, known to dissociate nNOS dimers into monomers) with concomitantly reduced levels of catalytically active dimers of nNOS in the PVN of rats with heart failure. To elucidate the molecular mechanism by which Angiotensin II (Ang II) increases PIN expression, we used Sprague-Dawley rats (250-300 g) subjected to intracerebroventricular infusion of Ang II (20 ng/min, 0.5 μl/h) or saline as vehicle (Veh) for 14 days through osmotic mini-pumps and NG108-15 hybrid neuronal cell line treated with Ang II as an in vitro model. Ang II infusion significantly increased baseline renal sympathetic nerve activity and mean arterial pressure. Ang II infusion increased the expression of PIN (1.24 ± 0.04* Ang II vs. 0.65 ± 0.07 Veh) with a concomitant 50% decrease in dimeric nNOS and PIN-Ub conjugates (0.73 ± 0.04* Ang II vs. 1.00 ± 0.03 Veh) in the PVN. Substrate-dependent ligase assay in cells transfected with pCMV-(HA-Ub)8 vector revealed a reduction of HA-Ub-PIN conjugates after Ang II and a proteasome inhibitor, Lactacystin (LC), treatment (4.5 ± 0.7* LC Ang II vs. 9.2 ± 2.5 LC). TUBE (Tandem Ubiquitin-Binding Entities) assay showed decrease PIN-Ub conjugates in Ang II-treated cells (0.82 ± 0.12* LC Ang II vs. 1.21 ± 0.06 LC) while AT1R blocker, Losartan (Los) treatment diminished the Ang II-mediated stabilization of PIN (1.21 ± 0.07 LC Los vs. 1.16 ± 0.04* LC Ang II Los). Taken together, our studies suggest that increased central levels of Ang II contribute to the enhanced expression of PIN leading to reduced expression of the dimeric form of nNOS, thus diminishing the inhibitory action of NO• on pre-autonomic neurons in the PVN resulting in increased sympathetic outflow.

The potency of magnetic resonance imaging (MRI) to measure the exact extent of osteonecrosis of the femoral head (ONFH) remains uncertain. The objective of this study was to determine if the volume of necrosis assessed with MRI accurately reflects the volume of architectural mineral alterations in osteonecrosis of the femoral head by comparison with high-resolution microfocus X-ray computed tomography (HR-μCT). Fourteen male patients aged 53 years [46.2;59.0] suffering from ONFH were prospectively enrolled to undergo preoperative MRI and ex vivo analysis using HR-μCT. The necrotic zone on T1-weighted MRI scans was defined as total necrosis (delimited by the low-signal peripheral band) or dark necrosis (low-signal lesions only). The HR-μCT scans delimited outer necrosis and inner necrosis by including or excluding the sclerotic zone. The intra-class correlation coefficient (ICC) was calculated to compare the agreement of surface areas and volumes of necrosis measurements with the two techniques. There was an overall excellent agreement between MRI dark necrosis volume and HR-μCT outer necrosis volume (ICC=0.91[0.54;0.98]) while the MRI total necrosis volume showed poor agreement with both HR-μCT delimitations of necrosis volume. For surface area, agreement between MRI dark necrosis and HR-μCT delimitations was good for inner necrosis (ICC=0.70[0.21;0.9]) and moderate for outer necrosis (ICC=0.58[0.07;0.85]). This study demonstrates that measurement of the MRI lesions provides a reliable assessment of the extent of ONFH-related architectural damage.

Pistachio hull has traditionally been used to treat peptic ulcer, hemorrhoids, oral and cutaneous wounds.

Nerve graft reconstruction of gap defects may result in poor clinical outcomes, particularly with long regeneration distances. Electrical stimulation (ES) of nerves may improve outcomes in such patients. A single session of ES at 20 Hz for 1 h significantly enhances axon regeneration in animals and human subjects after nerve crush or nerve transection and repair. The objectives of this study were to evaluate if ES enhances axon regeneration through nerve grafts and if there is added benefit of a second, delayed session of ES (serial ES) on axon regeneration as compared to a single session only of ES. In female rats, a gap defect was created in the hindlimb common peroneal (CP) nerve and immediately reconstructed with a 10 mm nerve autograft (Experiment 1) or a 20 mm nerve autograft (Experiment 2). In Experiment 1, rats were randomized to 1 h of CP nerve ES or sham stimulation. In Experiment 2, rats were randomized to control (sham ES + sham ES), single ES (ES + sham ES), or serial ES (ES + ES), which consisted of an initial 1 h session of either ES or sham stimulation of the CP nerve, followed by a second 1 h session of ES or sham stimulation of the CP nerve 4 weeks later. In both experiments, after a 6 week period of nerve regeneration, CP neurons that had regenerated axons distal to the autograft were retrograde labelled for enumeration, and the CP nerve distal to the autograft was harvested for histomorphometry. In Experiment 1, rats that received CP nerve ES had statistically significantly more motor (p < .05) and sensory (p < .05) neurons that regenerated axons distal to the 10 mm nerve autograft, with more myelinated axons on histomorphometry (p < .001). Similarly, in Experiment 2, significantly more motor (p < .01) and sensory (p < .05) neurons regenerated axons distal to the 20 mm nerve autograft after a single session or two sessions of CP nerve ES. There was no significant difference in the number of regenerated motor or sensory neurons between rats with 20 mm CP nerve autografts receiving either one or two sessions of CP nerve ES (p > .05). In conclusion, a single session of ES enhances axon regeneration following nerve autografting with no added effect of a second, delayed session of ES. These findings support previous studies in animals and humans of the robust effect of a single session of ES in promoting nerve regeneration following injury and repair.

Direct-acting antiviral (DAA) therapy for HCV has high efficacy and limited toxicity. We hypothesised that the efficacy of glecaprevir-pibrentasvir for chronic HCV with a simplified treatment monitoring schedule would be non-inferior to a standard treatment monitoring schedule.

Paeonia lactiflora Pall. has long been used to treat inflammatory skin diseases, such as psoriasis.

Analysis of human muscle diseases highlights the role of mitochondrial dysfunction in the skeletal muscle. Our previous work revealed that diverse upstream events correlated with altered mitochondrial proteome in human muscle biopsies. However, several proteins showed relatively unchanged expression suggesting that post-translational modifications, mainly protein phosphorylation could influence their activity and regulate mitochondrial processes. We conducted mitochondrial phosphoprotein profiling, by proteomics approach, of healthy human skeletal muscle (n = 10) and three muscle diseases (n = 10 each): Dysferlinopathy, Polymyositis and Distal Myopathy with Rimmed Vacuoles. Healthy human muscle mitochondrial proteins displayed 253 phosphorylation sites (phosphosites), which contributed to metabolic and redox processes and mitochondrial organization etc. Electron transport chain complexes accounted for 84 phosphosites. Muscle pathologies displayed 33 hyperphosphorylated and 14 hypophorphorylated sites with only 5 common proteins, indicating varied phosphorylation profile across muscle pathologies. Molecular modelling revealed altered local structure in the phosphorylated sites of Voltage-Dependent Anion Channel 1 and complex V subunit ATP5B1. Molecular dynamics simulations in complex I subunits NDUFV1, NDUFS1 and NDUFV2 revealed that phosphorylation induced structural alterations thereby influencing electron transfer and potentially altering enzyme activity. We propose that altered phosphorylation at specific sites could regulate mitochondrial protein function in the skeletal muscle during physiological and pathological processes.

Previously, we evaluated serum sialic acid (SA) levels in buffaloes naturally infected with T. annulata. In the current paper, we conducted a further study on oxidative/nitrosative stress biomarkers in erythrocyte lysate samples of the same buffaloes. DNA damage also was assessed. Additionally, we tested whether, there is any correlation between SA and the aforementioned indicators or not. To achieve these aims, several biomarkers including the activities of key antioxidant enzymes, superoxide dismutase, glutathione peroxidase and catalase, as well as malondialdehyde (MDA), protein carbonyl (PCO), nitric oxide contents (NO), total antioxidant capacity (TAC) and DNA damage levels were measured. The obtained results showed that the activities of the antioxidant enzymes and TAC levels decreased significantly as the percentage of parasitemia increased accordingly. Also, a significant increase in the levels of PCO, MDA, NO and DNA damage were recorded, depending on the degree of parasitemia. There was a significant correlation between oxidative/nitrosative stress indicators and SA. Conclusively, T. annulata infection in buffaloes is associated with a parasitic burden-dependent oxidative/nitrosative damages to erythrocytes and SA plays a crucial role in pathogenesis of the disease, as it is tightly correlated with oxidative/nitrosative indicators.

Neonatal encephalopathy frequently results from hypoxia-ischemia (HI) or inflammation in preterm or term neonates. Neuropathology depends on cerebral development at insult time, but the poor correlation of neuromotor, cognitive, and behavioral disabilities in infancy with initial imaging and clinical records precludes early prognosis. The Rice-Vannucci HI procedure was applied to wild type and tissue plasminogen activator knockout (tPA-KO) mice as surrogates for human preterm (with five-day-old postnatal (P5) mice) or human term (with ten-day-old postnatal (P10) mice). Acute and delayed T2-magnetic resonance imaging (T2-MRI) signals and cognitive deficits in adulthood (spatial memory and social interaction) were investigated in the same animals. Early vascular tPA and matrix metalloproteinase-9 (MMP-9) activities, blood-brain barrier permeability to water or IgG, and microglial activation were assessed separately. HI in P5 or P10 mice induced early hemisphere swelling in T2-MRI scans, and a delayed atrophy of the cortex and hippocampus, but affected white matter in the P5 group only, irrespective of the wild type or tPA-KO genotype. Adults had no motor disabilities, but we did find HI-induced age-dependent deficits, preferentially social interaction and activity in P5 mice, and spatial learning in P10 mice. In P5 mice, tPA-KO prevented MMP-9 activation, IgG extravasation, microglial activation, and behavior impairments. In P10 mice, MMP-9 activation and inflammatory processes remained in the hippocampus of the tPA-KO group, and also contributed to persistent spatial learning deficits. Perinatal HI in mice mimicked the unpredictability of outcomes from imaging in human clinics. Delayed deficits appeared associated to vascular dysfunction-induced inflammation, which recalls our previous work showing major vascular maturation between P5 and P10 stages. Using omics to explore neural, glial, or brain vessel markers in neonate blood may be a promising perspective to identify pertinent prognostic tools.

Synaptophysin (Syp) is an integral membrane protein of synaptic vesicles, and is ubiquitously expressed in neurons throughout the brain. As Syp expression is correlated with synaptogenesis during development of the central nervous system, the expression of Syp is considered to be a critical aspect of neuronal maturation and circuit formation. However, little information is available concerning the regulatory mechanisms of Syp gene expression during postnatal development of the brain. In the present study, we investigated changes in Syp mRNA in the hippocampus of mice during postnatal development, and examined the gene regulation mechanisms, focusing on DNA methylation. We found that hippocampal Syp expression involving both mRNA and protein levels increased during the first two weeks of life, and that this increase was accompanied by a transition from hypermethylation to hypomethylation at the CpG sites of the Syp gene upstream region. In addition, DNA demethylating agent 5-Aza-2'-deoxycytidine (5-aza-dC) de-repressed Syp gene expression both in vitro in Neuro-2a mouse neuronal cells and in vivo in the hippocampus of early postnatal mice. Furthermore, the methylation levels at upstream region of Syp gene in the hippocampus of developing mice was decreased by intraperitoneal injection of 5-aza-dC. These results suggest that Syp gene regulation, at least during postnatal brain development, could be mediated by DNA methylation. Our findings promote understanding of the molecular basis of synaptogenesis during postnatal brain development, and provide novel insight into therapeutic aspects of neurodevelopmental disorders involving synaptic dysfunction.

Lung cancer is one of the deadliest cancers worldwide. Late diagnosis at an advanced, inoperable stage makes chemotherapy a treatment of choice, yet, with low response rates. The hedgehog signaling pathway (HHSP) is often reactivated in cancer. We identified miR-182-5p as a regulator of GLI2, a transcriptional regulator of the HHSP, and explored the role of the miR-182-5p/GLI2 axis in carcinogenesis and cisplatin resistance of lung adenocarcinoma (LADC). Expression of miRNAs and target genes was analyzed by RT-qPCR, expression of the GLI-protein family in LADC and adjacent lung tissue (n = 27 pairs) by immunohistochemistry. MiR-182-5p was manipulated, and data were generated by immunoblotting, immunofluorescence, apoptosis, proliferation/viability, dual-luciferase-, and colony forming assays. MiR-182-5p was down-regulated in cisplatin-resistant LADC cells and directly targeted GLI2. Interference with miR-182-5p or GLI2 silencing resulted in modulation of cell proliferation, clonogenic potential, and cisplatin-sensitivity. HHSP was markedly reactivated in LADC tissue compared to adjacent non-malignant lung tissue. Our results indicate that the miR-182-5p/GLI2 axis modulates tumorigenesis and cisplatin-resistance in LADC cells, by influencing the HHSP. Therefore, this axis might be considered as a potential biomarker and future therapeutic target in LADC patients.