Conformational and Electronic Variations in 1,2- and 1,5a-Cyclophellitols and their Impact on Retaining α-Glucosidase Inhibition.

Glycoside hydrolases (glycosidases) take part in myriad biological processes and are important therapeutic targets. Competitive and mechanism-based inhibitors are useful tools to dissect their biological role and comprise a good starting point for drug discovery. The natural product, cyclophellitol, a mechanism-based, covalent and irreversible retaining ß-glucosidase inhibitor has inspired the design of diverse α- and ß-glycosidase inhibitor and activity-based probe scaffolds. Here, we sought to deepen our understanding of the structural and functional requirements of cyclophellitol-type compounds for effective human α-glucosidase inhibition. We synthesized a comprehensive set of α-configured 1,2- and 1,5a-cyclophellitol analogues bearing a variety of electrophilic traps. The inhibitory potency of these compounds was assessed towards both lysosomal and ER retaining α-glucosidases. These studies revealed the 1,5a-cyclophellitols to be the most potent retaining α-glucosidase inhibitors, with the nature of the electrophile determining inhibitory mode of action (covalent or non-covalent). DFT calculations support the ability of the 1,5a-cyclophellitols, but not the 1,2-congeners, to adopt conformations that mimic either the Michaelis complex or transition state of α-glucosidases.

Recapitulation of Skewed X-Inactivation in Female Ornithine Transcarbamylase-Deficient Primary Human Hepatocytes in the FRG Mouse: A Novel System for Developing Epigenetic Therapies.

Realization of the immense therapeutic potential of epigenetic editing requires development of clinically predictive model systems that faithfully recapitulate relevant aspects of the target disease pathophysiology. In female patients with ornithine transcarbamylase (OTC) deficiency, an X-linked condition, skewed inactivation of the X chromosome carrying the wild-type OTC allele is associated with increased disease severity. The majority of affected female patients can be managed medically, but a proportion require liver transplantation. With rapid development of epigenetic editing technology, reactivation of silenced wild-type OTC alleles is becoming an increasingly plausible therapeutic approach. Toward this end, privileged access to explanted diseased livers from two affected female infants provided the opportunity to explore whether engraftment and expansion of dissociated patient-derived hepatocytes in the FRG mouse might produce a relevant model for evaluation of epigenetic interventions. Hepatocytes from both infants were successfully used to generate chimeric mouse-human livers, in which clusters of primary human hepatocytes were either OTC positive or negative by immunohistochemistry (IHC), consistent with clonal expansion from individual hepatocytes in which the mutant or wild-type OTC allele was inactivated, respectively. Enumeration of the proportion of OTC-positive or -negative human hepatocyte clusters was consistent with dramatic skewing in one infant and minimal to modest skewing in the other. Importantly, IHC and fluorescence-activated cell sorting analysis of intact and dissociated liver samples from both infants showed qualitatively similar patterns, confirming that the chimeric mouse-human liver model recapitulated the native state in each infant. Also of importance was the induction of a treatable metabolic phenotype, orotic aciduria, in mice, which correlated with the presence of clonally expanded OTC-negative primary human hepatocytes. We are currently using this unique model to explore CRISPR-dCas9-based epigenetic targeting strategies in combination with efficient adeno-associated virus (AAV) gene delivery to reactivate the silenced functional OTC gene on the inactive X chromosome.

The byssal-producing glands and proteins of the silverlip pearl oyster Pinctada maxima (Jameson, 1901).

Pinctada maxima are most well known for their production of high-quality natural pearls. They also generate another natural material, the byssus, an adhesive thread critical for steadfast attachment underwater. Herein, P. maxima byssal threads were analysed via proteotranscriptomics to reveal 49 proteins. Further characterisation was undertaken on five highly expressed genes: glycine-rich thread protein (GRT; also known as PUF3), apfp1/perlucin-like protein (Pmfp1); peroxidase; thrombospondin 1, and Balbiani ring 3 (BR3), which showed localised tissue expression. The spatial distribution of GRT and Pmfp1 via immunodetection combined with histology helped to identify glandular regions of the foot that contribute to byssal thread production: the byssal gland, the duct gland, and two thread-forming glands of basophilic and acidophilic serous-like cells. This work advanced primary knowledge on the glands involved in the creation of byssal threads and the protein composition of the byssus for P. maxima, providing a platform for the design of marine biopolymers.

[Common foundation of alexithymia and expressive suppression]. / Gemeinsame Grundlage von Alexithymie und expressiver Suppression: Ergebnisse einer empirischen Studie.

BACKGROUND: International studies could already prove a correlation between alexithymia and expressive suppression. This relationship has only been marginally considered in the German literature so far. The prioritized aim of the present study was to investigate a correlative and factorial relationship between alexithymia and expressive suppression. MATERIAL AND METHODS: A total of 317 persons participated in an online survey. Data on alexithymia and expressive suppression were collected using the German versions of the Toronto alexithymia scale (TAS-26) and the emotion regulation questionnaire (ERQ). RESULTS: The results showed highly significant correlations between the TAS-26 subscale "difficulty in identifying feelings" and the ERQ scale "suppression" (r = 0.5; p < 0.001) and between the TAS-26 subscale "difficulty in describing feelings" and the ERQ scale "suppression" (r = 0.64; p < 0.001). The results of an exploratory factor analysis revealed a two-factor solution with common factor for the TAS-26 subscales "difficulty in identifying feelings" and "difficulty in describing feelings" and the ERQ scale "suppression" with a common variance of 38.2% (χ2 = 363.843, p < 0.001, Kaiser-Meyer-Olkin, KMO, value = 0.699). CONCLUSION: The results indicate that the TAS-26 scales in the components "difficulty in identifying feelings" and "difficulty in describing feelings" and the ERQ scale "suppression" in the component of "expressive suppression" have a common construct, which is referred to with the term speechlessness.

Myocarditis and Subclinical-Like Infection Associated With SARS-CoV-2 in Two Cats Living in the Same Household in France: A Case Report With Literature Review.

This report provides the first clinical, radiographic, echocardiographic, and biological description of SARS-CoV-2-associated myocarditis with a 6-month follow-up in a 5-year-old obese male domestic shorthair cat (Cat-1) presented for refractory congestive heart failure, with high cardiac troponin-I level (5.24 ng/ml), and a large lingual ulcer. The animal was SARS-CoV-2 positive on serology. The other cat living in the same household (Cat-2) never showed any clinical sign but was also confirmed SARS-CoV-2 positive on serology. Both cats were SARS-CoV-2 PCR negative. Cat-1 had closer contact than Cat-2 with their owner, who had been in close contact with a coworker tested PCR positive for COVID-19 (Alpha (B.1.1.7) variant) 4 weeks before Cat-1's first episode of congestive heart failure. A focused point-of-care echocardiography at presentation revealed for Cat-1 numerous B-lines, pleural effusion, severe left atrial dilation and dysfunction, and hypertrophic cardiomyopathy phenotype associated with focal pulmonary consolidations. Both myocarditis and pneumonia were suspected, leading to the prescription of cardiac medications and antibiotics. One month later, Cat-1 recovered, with normalization of left atrial size and function, and radiographic and echocardiography disappearance of heart failure signs and pulmonary lesions. An extensive literature review of SARS-CoV-2-related cardiac injury in pets in comparison with human pathology is discussed.

Non-reciprocity in optical fiber links: experimental evidence.

Fundamental limits of fiber link are set by non-reciprocal effects that violate the hypothesis of equality between forward and backward path. Non-reciprocal noise arises technically from the set-up asymmetry, and fundamentally by the Sagnac effect when the fiber link encloses a non-zero area. As a pre-requisite for observation of Sagnac effect in fiber links, we present a study on phase noise and frequency stability contributions affecting coherent optical frequency transfer in bi-directional fiber links. Both technical and fundamental limitations of Two-Way optical frequency transfer are discussed. Our model predicts and our experiments substantially verify that the dominant noise mechanism at low Fourier frequencies is the polarization asymmetry induced by the temperature and relative humidity variations impacted on fiber links. The flicker noise floor due to the non-reciprocal noise arising from polarization mode dispersion is evidenced for the first time. We perform a post-processing approach which enables us to remove this polarization noise, improve the long-term stability and remove a frequency bias. We evaluate the uncertainty contributions of all the effects discussed for our 50 km spooled fiber link, dominated by its non-reciprocal noise induced by polarization mode dispersion with uncertainty of 1.9( ± 0.8)( ± 1.2) × 10-20. After correction, the linear drift of the residual phase is as low as 27 yoctosecond/s, leading to an uncertainty of the frequency transfer of 2.6 ( ± 39) × 10-22, confirming its potential for searching for more fundamental effects such as Sagnac effect or transient frequency variation due to dark matter.

The protein and volatile components of trail mucus in the Common Garden Snail, Cornu aspersum.

The Common or Brown Garden Snail, Cornu aspersum, is an invasive land snail that has successfully colonized a diverse range of global environments. Like other invasive land snails, it is a significant pest of a variety of agricultural crops, including citrus, grapes and canola. Cornu aspersum secretes a mucus trail when mobile that facilitates locomotion. The involvement of the trail in conspecific chemical communication has also been postulated. Our study found that anterior tentacle contact with conspecific mucus elicited a significant increase in heart rate from 46.9 to 51 beats per minute. In order to gain a better understanding of the constituents of the trail mucus and the role it may play in snail communication, the protein and volatile components of mucus trails were investigated. Using two different protein extraction methods, mass spectrometry analysis yielded 175 different proteins, 29 of which had no significant similarity to any entries in the non-redundant protein sequence database. Of the mucus proteins, 22 contain features consistent with secreted proteins, including a perlucin-like protein. The eight most abundant volatiles detected using gas chromatography were recorded (including propanoic acid and limonene) and their potential role as putative pheromones are discussed. In summary, this study has provided an avenue for further research pertaining to the role of trail mucus in snail communication and provides a useful repository for land snail trail mucus components. This may be utilized for further research regarding snail attraction and dispersal, which may be applied in the fields of agriculture, ecology and human health.

The P450 multigene family of Fontainea and insights into diterpenoid synthesis.

BACKGROUND: Cytochrome P450s (P450s) are enzymes that play critical roles in the biosynthesis of physiologically important compounds across all organisms. Although they have been characterised in a large number of plant species, no information relating to these enzymes are available from the genus Fontainea (family Euphorbiaceae). Fontainea is significant as the genus includes species that produce medicinally significant epoxy-tigliane natural products, one of which has been approved as an anti-cancer therapeutic. RESULTS: A comparative species leaf metabolome analysis showed that Fontainea species possess a chemical profile different from various other plant species. The diversity and expression profiles of Fontainea P450s were investigated from leaf and root tissue. A total of 103 and 123 full-length P450 genes in Fontainea picrosperma and Fontainea venosa, respectively (and a further 127/125 partial-length) that were phylogenetically classified into clans, families and subfamilies. The majority of P450 identified are most active within root tissue (66.2% F. picrosperma, 65.0% F. venosa). Representatives within the CYP71D and CYP726A were identified in Fontainea that are excellent candidates for diterpenoid synthesis, of which CYP726A1, CYP726A2 and CYP71D1 appear to be exclusive to Fontainea species and were significantly more highly expressed in root tissue compared to leaf tissue. CONCLUSION: This study presents a comprehensive overview of the P450 gene family in Fontainea that may provide important insights into the biosynthesis of the medicinally significant epoxy-tigliane diterpenes found within the genus.

Expanded Neonatal Bloodspot Screening Programmes: An Evaluation Framework to Discuss New Conditions With Stakeholders.

Neonatal bloodspot screening (NBS) programmes that screen for rare but serious conditions are expanding worldwide. Fast developments for testing and treatment put pressure on implementation processes. In 2015 the Netherlands embarked on an NBS expansion from 17 to 31 conditions. An evaluation framework was developed based on international NBS frameworks to gain insight in test properties, clinical findings, follow-up and implementation. A stakeholder process took place with implications for the planning of the expanded NBS panel. The evaluation framework progressed into a go/no go framework to start national screening, and is currently explored as basis for continuous evaluation of the NBS panel. The framework and stakeholder process may serve as an example for other programmes.

Evaluation of Mathematical Arterialization of Venous Blood in Intensive Care and Pulmonary Ward Patients.

BACKGROUND: Arterial blood gases are important when assessing acute or critically ill patients. Capillary blood and mathematical arterialization of venous blood have been proposed as alternative methods, eliminating pain and complications of arterial puncture. OBJECTIVES: This study compares the arterial samples, arterialized venous samples, and capillary samples in ICU and pulmonary ward patients. METHOD: Ninety-one adult patients with respiratory failure were included in the analysis. Arterial, peripheral venous, and mathematically arterialized venous samples were compared in all patients using Bland-Altman analysis, with capillary samples included in 36 patients. RESULTS: Overall for pH and PCO2, arterialized venous values, and in the subset of 36 patients, capillary values, compared well to arterial values and were within the pre-defined clinically acceptable differences (pH ± 0.05 and PCO2 ± 0.88 kPa). For PO2, arterialized or capillary values describe arterial with similar precision (PO2 arterialized -0.03, LoA -1.48 to 1.42 kPa and PO2 capillary 0.82, LoA -1.36 to 3 kPa), with capillary values underestimating arterial. CONCLUSIONS: Mathematical arterialization functions well in a range of patients in an ICU and ward outside the country of development of the method. Furthermore, accuracy and precision are similar to capillary blood samples. When considering a replacement for arterial sampling in ward patients, using capillary sampling or mathematical arterialization should depend on logistic ease of implementation and use rather than improved measurements of using either technique.

Unidirectional two-way optical frequency comparison and its fundamental limitations.

High-quality frequency transfer based on existing telecommunication fiber links allows state-of-the-art microwave and optical clocks to be compared on a continental and potentially even intercontinental scale. We present a half-a-year-long data set of unidirectional optical frequency transfer over a 2×43km urban fiber link. We observe a relative frequency instability of 8.0×10-16 at 1 s and at best 6.0×10-18 at 106s integration time. Our results show that the unidirectional two-way method gives the possibility to perform comparisons of international primary standards over fiber links faster than over satellite links. Moreover, we investigate the major limiting factors of such a unidirectional setup.

Reciprocity of propagation in optical fiber links demonstrated to 10-21.

We present a study of the fundamental limit of fiber links using dedicated link architecture. We use an experimental arrangement that enables us to detect the forward and backward propagation noise independently and simultaneously in optical fiber and where the optical phase evolution is expected to be driven by the only contribution of the reference arms of the Michelson interferometer ensemble. In this article, we demonstrate indeed the high correlation between the optical phase evolution and the temperature variation of the interferometer ensemble, leading to a frequency offset of (4.4±2.3)×10-21. Using a simple temperature model and a Bayesian analysis to evaluate the model parameters, we show that the temperature effect can be compensated with post-processing, removing the frequency offset down to (0.5±2.0)×10-21. The residual slope of the optical phase evolution over 33 days is 350 yoctosecond/s. Using a global temperature parameter, we divide these 33 days dataset in four subsets and analyse their uncertainties. We show that they are self-consistent when the temperature is taken into account. This provides an alternative method to evaluate the accuracy of a fiber link, especially when the dataset includes large dead times. The result is finally interpreted as a test of the reciprocity of the propagation delay in an optical fiber. This unprecedented transfer capability could enable the comparisons of future optical clocks with expected performance at 10-20 level and open new possibilities for stringent tests of special and general relativity.

Multi-omics investigations within the Phylum Mollusca, Class Gastropoda: from ecological application to breakthrough phylogenomic studies.

Gastropods are the largest and most diverse class of mollusc and include species that are well studied within the areas of taxonomy, aquaculture, biomineralization, ecology, microbiome and health. Gastropod research has been expanding since the mid-2000s, largely due to large-scale data integration from next-generation sequencing and mass spectrometry in which transcripts, proteins and metabolites can be readily explored systematically. Correspondingly, the huge data added a great deal of complexity for data organization, visualization and interpretation. Here, we reviewed the recent advances involving gastropod omics ('gastropodomics') research from hundreds of publications and online genomics databases. By summarizing the current publicly available data, we present an insight for the design of useful data integrating tools and strategies for comparative omics studies in the future. Additionally, we discuss the future of omics applications in aquaculture, natural pharmaceutical biodiscovery and pest management, as well as to monitor the impact of environmental stressors.

Studying the fundamental limit of optical fiber links to the 10-21 level.

We present a hybrid fiber link combining effective optical frequency transfer and evaluation of performances with a self-synchronized two-way comparison. It enables us to detect the round-trip fiber noise and each of the forward and backward one-way fiber noises simultaneously. The various signals acquired with this setup allow us to study quantitatively several properties of optical fiber links. We check the reciprocity of the accumulated noise forth and back over a bi-directional fiber to the level of 3.1(±3.9) × 10-20 based on a 160000s continuous data. We also analyze the noise correlation between two adjacent fibers and show the first experimental evidence of interferometric noise at very low Fourier frequency. We estimate redundantly and consistently the stability and accuracy of the transferred optical frequency over 43 km at 4 × 10-21 level after 16 days of integration and demonstrate that a frequency comparison with instability as low as 8 × 10-18 would be achievable with uni-directional fibers in urban area.

Rituximab impedes natural killer cell function in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis patients: A pilot in vitro investigation.

BACKGROUND: A recent in vitro pilot investigation reported Rituximab significantly reduced natural killer (NK) cell cytotoxicity in healthy donors. Chronic fatigue syndrome/Myalgic encephalomyelitis (CFS/ME) is a debilitating disorder of unknown etiology. A consistent finding is a significant reduction in NK cell cytotoxicity. Rituximab has been reported having questionable potential therapeutic benefits for the treatment of CFS/ME, however, the potential effects of Rituximab on NK cell cytotoxicity in CFS/ME patients are yet to be determined. METHODS: A total of eight CFS/ME patients (48.63 ± 15.69 years) and nine non-fatigued controls (NFC) (37.56 ± 11.06 years) were included using the Fukuda case definition. Apoptotic function, lytic proteins and degranulation markers were measured on isolated NK cells using flow cytometry following overnight incubation with Rituximab at 10 µg/ml and 100 µg/ml. RESULTS: There was a significant reduction in NK cell lysis between CFS/ME patients and NFC following incubation with Rituximab at 100 µg/ml at 12.5:1 and 6.25:1 effecter-target (E:T) ratios (p < 0.05). However, there was no significant difference for NFC following incubation with Rituximab at 10 µg/ml and 100 µg/ml. There was no significant difference between CFS/ME patients and NFC for granzyme A and granzyme B prior to incubation with Rituximab and following overnight incubation with Rituximab at 10 µg/ml. There was a significant decrease in granzyme B in CFS/ME patients compared to NFC with 100 µg/ml of Rituximab prior to K562 cells stimulation (p < 0.05). There was a significant increase in CD107a (p < 0.05) and CD107b expression (p < 0.01) in NFC after stimulation with K562 cells prior to incubation with Rituximab. There was a significant increase in CD107b expression between CFS/ME patients and NFC prior to incubation with Rituximab and without stimulation of K562 cells (p < 0.01). Importantly, there was a significant increase in CD107b following overnight incubation with 100 µg/ml of Rituximab in NFC prior to K562 cells stimulation (p < 0.01). CONCLUSION: This study reports significant decreases in NK cell lysis and a significant increase in NK cell degranulation following Rituximab incubation in vitro in CFS/ME patients, suggesting Rituximab may be toxic for NK cells. Caution should be observed in clinical trials until further investigations in a safe and controlled in vitro setting are completed.

A targeted genome association study examining transient receptor potential ion channels, acetylcholine receptors, and adrenergic receptors in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis.

BACKGROUND: Chronic Fatigue Syndrome, also known as Myalgic Encephalomyelitis (CFS/ME) is a debilitating condition of unknown aetiology. It is characterized by a range of physiological effects including neurological, sensory and motor disturbances. This study examined candidate genes for the above clinical manifestations to identify single nucleotide polymorphism (SNP) alleles associated with CFS/ME compared with healthy controls. METHODS: DNA was extracted and whole genome genotyping was performed using the HumanOmniExpress BeadChip array. Gene families for transient receptor potential ion channels, acetylcholine receptors, and adrenergic receptors, and acetylcholinesterase were targeted. The frequency of each SNP and their association between CFS/ME and healthy controls was examined using Fisher's exact test, and to adjust for multiple testing, False Detection Rate (FDR) and Bonferroni corrections were applied (p < 0.05). RESULTS: The study included 172 participants, consisting of 95 Fukuda defined CFS/ME patients (45.8 ± 8.9; 69 % female) and 77 healthy controls (42.3 ± 10.3; 63 % female). A total of 950 SNPs were included for analysis. 60 significant SNPs were associated with CFS/ME compared with healthy controls. After applying FDR and Bonferroni corrections, SNP rs2322333 in adrenergic receptor α1 (ADRA1A) was higher in CFS/ME compared with healthy controls (45.3 % vs. 23.4 %; p = 0.059). The genotype class that was homozygous minor (AA) was substantially lower in CFS/ME compared with healthy controls (4.2 % vs. 24.7 %). CONCLUSIONS: This study reports for the first time the identification of ADRA1A and a possible association between CFS/ME and genotype classes. Further examination of the functional role of this class of adrenergic receptors may elucidate the cause of particular clinical manifestations observed in CFS/ME.

Impact of rapid influenza diagnostic test on physician estimation of viral infection probability in paediatric emergency department during epidemic period.

BACKGROUND: The clinical diagnosis of influenza is difficult in the younger children. OBJECTIVES: Evaluate the impact of rapid influenza diagnostic test (RIDT) on clinicians' estimation of the clinical probability of influenza in children. STUDY DESIGN: This prospective study included children aged from 1 month to 5 years who were admitted in a university paediatric emergency department during an influenza epidemic period and presented with fever without source. The RIDT Quickvue(®) was performed on nasopharyngeal aspiration and results were confirmed with immunofluorescence and/or PCR. The clinical probability of influenza and serious bacterial infection (SBI) was evaluated for each child before and after the physician(s) was informed of the RIDT results. RESULTS: 170 children were included from January 15th through March 18th, 2013. After the only clinical examination, the overall clinical probability of influenza was 66.0% [CI 95%: 63.04-68.4], and was significantly increased at 92.4% [CI 95%: 89.5-95.3] in case of positive RIDT and significantly decreased at 30.8% [CI 95%: 29.0-32.5] in case of negative RIDT without knowing the results of laboratory tests. Whereas the initial clinical probability of influenza were appropriate regarding the prevalence (66.0% vs. 57.0%), the probability of SBI was overestimated (30.2% vs. 8.8%). The RIDT result positive enabled a significant decrease in orders for chest X-rays (64,4% vs. 45.8%, p<0,05) and laboratory tests (71,1% vs. 41.1%, p<0,05). CONCLUSIONS: The RIDT seems to be a useful diagnostic tool for ED clinicians in epidemic conditions. Improving clinician estimation of flu probability would reduce orders for imaging and testing.

Interoception, contemplative practice, and health.

Interoception can be broadly defined as the sense of signals originating within the body. As such, interoception is critical for our sense of embodiment, motivation, and well-being. And yet, despite its importance, interoception remains poorly understood within modern science. This paper reviews interdisciplinary perspectives on interoception, with the goal of presenting a unified perspective from diverse fields such as neuroscience, clinical practice, and contemplative studies. It is hoped that this integrative effort will advance our understanding of how interoception determines well-being, and identify the central challenges to such understanding. To this end, we introduce an expanded taxonomy of interoceptive processes, arguing that many of these processes can be understood through an emerging predictive coding model for mind-body integration. The model, which describes the tension between expected and felt body sensation, parallels contemplative theories, and implicates interoception in a variety of affective and psychosomatic disorders. We conclude that maladaptive construal of bodily sensations may lie at the heart of many contemporary maladies, and that contemplative practices may attenuate these interpretative biases, restoring a person's sense of presence and agency in the world.

Cascaded optical fiber link using the internet network for remote clocks comparison.

We report a cascaded optical link of 1100 km for ultra-stable frequency distribution over an Internet fiber network. The link is composed of four spans for which the propagation noise is actively compensated. The robustness and the performance of the link are ensured by five fully automated optoelectronic stations, two of them at the link ends, and three deployed on the field and connecting the spans. This device coherently regenerates the optical signal with the heterodyne optical phase locking of a low-noise laser diode. Optical detection of the beat-note signals for the laser lock and the link noise compensation are obtained with stable and low-noise fibered optical interferometer. We show 3.5 days of continuous operation of the noise-compensated 4-span cascaded link leading to fractional frequency instability of 4x10(-16) at 1-s measurement time and 1x10(-19) at 2000 s. This cascaded link was extended to 1480-km with the same performance. This work is a significant step towards a sustainable wide area ultra-stable optical frequency distribution and comparison network at a very high level of performance.

Ultra-stable long distance optical frequency distribution using the Internet fiber network.

We report an optical link of 540 km for ultrastable frequency distribution over the Internet fiber network. The stable frequency optical signal is processed enabling uninterrupted propagation on both directions. The robustness and the performance of the link are enhanced by a cost effective fully automated optoelectronic station. This device is able to coherently regenerate the return optical signal with a heterodyne optical phase locking of a low noise laser diode. Moreover the incoming signal polarization variation are tracked and processed in order to maintain beat note amplitudes within the operation range. Stable fibered optical interferometer enables optical detection of the link round trip phase signal. The phase-noise compensated link shows a fractional frequency instability in 10 Hz bandwidth of 5 × 10(-15) at one second measurement time and 2 × 10(-19) at 30,000 s. This work is a significant step towards a sustainable wide area ultrastable optical frequency distribution and comparison network.