The effect of cumulative energy in repeated subliminal transscleral cyclophotocoagulation: a retrospective study.

BACKGROUND: The long-term safety and efficacy of repeated applications of subliminal transscleral cyclophotocoagulation (SL-TSCPC) with a focus on cumulative energy was evaluated in glaucoma patients. METHODS: In this retrospective, multicentric study the data of a total of 82 eyes with various causes of glaucoma that were treated with a single or multiple applications of SL-TSCPC were collected. Treatments were performed under general or local anesthesia with an 810 nm diode laser. Power was 2000 mW; duty cycle, 31.3%; total treatment duration, 80-320 s; equaling a total energy of 50-200 J per treatment session. Fifty-five eyes (55 patients) presented for all follow-ups, and these eyes were selected for further statistical analysis. The mean age was 60.0 ± 17.1 years, and 22 (40%) of the patients were female. Intraocular pressure (IOP) and dependence on further glaucoma medication were evaluated at 12 months following the initial treatment. RESULTS: Eyes underwent 1 or 2 consecutive SL-TSCPC treatments. Median (min-max) baseline IOP of 34 (13-69) decreased to 21.5 (7-61), 22 (8-68), 20 (9-68), and 19.5 (3-60) mmHg at the 1, 3, 6, and 12-month postoperative timepoints respectively. The mean (± SD) IOP decrease at 12 months was 26 ± 27%, 39 ± 32%, and 49 ± 33% in the low (below 120 J, n = 18), medium (120-200 J, n = 24), and high (above 200 J, n = 13) cumulative energy groups respectively. At the 12-month timepoint, oral carbonic anhydrase use was discontinued in ¾ of the cases. CONCLUSIONS: It was found that the repeated application of SL-TSCPC safely and efficiently decreases IOP in a Caucasian population with heterogenous causes of glaucoma, eyes with silicone oil responded to a greater extent. Inclusion of cumulative energy scales may contribute to better addressing repeated procedures in a standardized fashion.

Glial cells LAP axon fragments.

In the nervous system, dead cell-derived material arising from injuries and neurodegeneration is normally removed by the phagocytic activity of macrophages or glia. Failure in this process can lead to excessive inflammation and secondary neurodegeneration. During phagocytosis, engulfed material is captured into phagosomes. Maturation and subsequent fusion of these vesicles with lysosomes may utilize components of the macroautophagy pathway that has been referred to as LC3-associated phagocytosis or LAP for short.

LC3-associated phagocytosis promotes glial degradation of axon debris after injury in Drosophila models.

Glial engulfment of neuron-derived debris after trauma, during development, and in neurodegenerative diseases supports nervous system functions. However, mechanisms governing the efficiency of debris degradation in glia have remained largely unexplored. Here we show that LC3-associated phagocytosis (LAP), an engulfment pathway assisted by certain autophagy factors, promotes glial phagosome maturation in the Drosophila wing nerve. A LAP-specific subset of autophagy-related genes is required in glia for axon debris clearance, encoding members of the Atg8a (LC3) conjugation system and the Vps34 lipid kinase complex including UVRAG and Rubicon. Phagosomal Rubicon and Atg16 WD40 domain-dependent conjugation of Atg8a mediate proper breakdown of internalized axon fragments, and Rubicon overexpression in glia accelerates debris elimination. Finally, LAP promotes survival following traumatic brain injury. Our results reveal a role of glial LAP in the clearance of neuronal debris in vivo, with potential implications for the recovery of the injured nervous system.

Despite its sequence identity with canonical H4, Drosophila H4r product is enriched at specific chromatin regions.

Histone variants are different from their canonical counterparts in structure and are encoded by solitary genes with unique regulation to fulfill tissue or differentiation specific functions. A single H4 variant gene (His4r or H4r) that is located outside of the histone cluster and gives rise to a polyA tailed messenger RNA via replication-independent expression is preserved in Drosophila strains despite that its protein product is identical with canonical H4. In order to reveal information on the possible role of this alternative H4 we epitope tagged endogenous H4r and studied its spatial and temporal expression, and revealed its genome-wide localization to chromatin at the nucleosomal level. RNA and immunohistochemistry analysis of H4r expressed under its cognate regulation indicate expression of the gene throughout zygotic and larval development and presence of the protein product is evident already in the pronuclei of fertilized eggs. In the developing nervous system a slight disequibrium in H4r distribution is observable, cholinergic neurons are the most abundant among H4r-expressing cells. ChIP-seq experiments revealed H4r association with regulatory regions of genes involved in cellular stress response. The data presented here indicate that H4r has a variant histone function.

Loss of ubiquitinated protein autophagy is compensated by persistent cnc/NFE2L2/Nrf2 antioxidant responses.

SQSTM1/p62-type selective macroautophagy/autophagy receptors cross-link poly-ubiquitinated cargo and autophagosomal LC3/Atg8 proteins to deliver them for lysosomal degradation. Consequently, loss of autophagy leads to accumulation of polyubiquitinated protein aggregates that are also frequently seen in various human diseases, but their physiological relevance is incompletely understood. Here, using a genetically non-redundant Drosophila model, we show that specific disruption of ubiquitinated protein autophagy and concomitant formation of polyubiquitinated aggregates has hardly any effect on bulk autophagy, proteasome activity and fly healthspan. We find that accumulation of ref(2)P/SQSTM1 due to a mutation that disrupts its binding to Atg8a results in the co-sequestering of Keap1 and thus activates the cnc/NFE2L2/Nrf2 antioxidant pathway. These mutant flies have increased tolerance to oxidative stress and reduced levels of aging-associated mitochondrial superoxide. Interestingly, ubiquitin overexpression in ref(2)P point mutants prevents the formation of large aggregates and restores the cargo recognition ability of ref(2)P, although it does not prevent the activation of antioxidant responses. Taken together, potential detrimental effects of impaired ubiquitinated protein autophagy are compensated by the aggregation-induced antioxidant response.Abbreviations: Atg8a: Autophagy-related 8a; cnc: cap-n-collar; IFM: indirect flight muscle; KEAP1: kelch like ECH associated protein 1; LIR: LC3-interacting region; NFE2L2/Nrf2: NFE2 like bZIP transcription factor 2; PB1: Phox and Bem1; ref(2)P: refractory to sigma P; SAR: selective autophagy receptor; UBA: ubiquitin-associated.

Experimental comparison of fiber optic parametric, Raman and erbium amplifiers for burst traffic for extended reach PONs.

We experimentally compare the performance of a polarization-independent fiber optic parametric amplifier (FOPA), a discrete Raman amplifier and a commercial erbium doped fiber amplifier (EDFA) for burst traffic amplification in extended reach passive optical networks (PON). We demonstrate that EDFA and Raman amplifiers suffer from severe transient effects, causing penalty on receiver sensitivity >5 dB for traffic bursts of 10 Gbps on-off keying signal shorter than 10 µs. On the other hand, we demonstrate that FOPA does not introduce a penalty on receiver sensitivity when amplifying signal bursts as short as 5 µs as compared to a non-burst signal. Therefore, FOPA used as a drop-in replacement for an EDFA or Raman amplifier allows us to improve receiver sensitivity by >3 dB for short signal bursts. We conclude that FOPA allows substantially increased power budget for an extended reach PON transmitting variable duration bursts. In addition, we identify the maximum burst duration tolerated by each examined amplifier.

2-D Materials for Ultrascaled Field-Effect Transistors: One Hundred Candidates under the Ab Initio Microscope.

Due to their remarkable properties, single-layer 2-D materials appear as excellent candidates to extend Moore's scaling law beyond the currently manufactured silicon FinFETs. However, the known 2-D semiconducting components, essentially transition metal dichalcogenides, are still far from delivering the expected performance. Based on a recent theoretical study that predicts the existence of more than 1800 exfoliable 2-D materials, we investigate here the 100 most promising contenders for logic applications. Their current versus voltage characteristics are simulated from first-principles, combining density functional theory and advanced quantum transport calculations. Both n- and p-type configurations are considered, with gate lengths ranging from 15 down to 5 nm. From this large collection of electronic materials, we identify 13 compounds with electron and hole currents potentially much higher than those in future Si FinFETs. The resulting database widely expands the design space of 2-D transistors and provides original guidelines to the materials and device engineering community.

Winter-Active Spider Fauna is Affected by Plantation Forest Type.

Plantations of non-native trees for commercial use are common practice in Europe. They are known to have severe ecological impacts on arthropod fauna by altering microclimatic conditions and reducing microhabitat diversity. However, the effect of plantation tree species on winter-active fauna is relatively unknown. Spiders are a diverse predatory arthropod taxon with strong effect on their prey populations. The composition of spider communities sensitively indicates changes in habitat structure. We established 40 sampling sites in five non-native pine and five native poplar plantations and collected spiders with pitfall traps for two winters in the Southern part of Hungary. We assessed the average height of vegetation and percentage cover of leaf litter, mosses, herbaceous vegetation, and shrubs to characterize habitat structure. We found species richness and activity density of spiders in the non-native compared to the native plantations, presumably due to the more temperate microclimate in pine than in poplar plantations. However, there was no significant effect of habitat structure and its interaction with forest type on species richness and activity density of spiders. Species composition of non-native and native plantation forests differed significantly. Furthermore, we identified six characteristic spider species of non-native plantations with preference for relatively moist habitat conditions. The single characteristic species, (Agroeca cuprea Menge, 1873) for the native plantations preferred dry and partly shaded habitats. We conclude that the effect of microclimatic differences and prey availability presumably overrides the effect of habitat structure on winter-active spiders.

One-Dimensional Edge Contacts to a Monolayer Semiconductor.

Integration of electrical contacts into van der Waals (vdW) heterostructures is critical for realizing electronic and optoelectronic functionalities. However, to date no scalable methodology for gaining electrical access to buried monolayer two-dimensional (2D) semiconductors exists. Here we report viable edge contact formation to hexagonal boron nitride (hBN) encapsulated monolayer MoS2. By combining reactive ion etching, in situ Ar+ sputtering and annealing, we achieve a relatively low edge contact resistance, high mobility (up to ∼30 cm2 V-1 s-1) and high on-current density (>50 µA/µm at VDS = 3V), comparable to top contacts. Furthermore, the atomically smooth hBN environment also preserves the intrinsic MoS2 channel quality during fabrication, leading to a steep subthreshold swing of 116 mV/dec with a negligible hysteresis. Hence, edge contacts are highly promising for large-scale practical implementation of encapsulated heterostructure devices, especially those involving air sensitive materials, and can be arbitrarily narrow, which opens the door to further shrinkage of 2D device footprint.

Electron Transport through Metal/MoS2 Interfaces: Edge- or Area-Dependent Process?

In ultrathin two-dimensional (2-D) materials, the formation of ohmic contacts with top metallic layers is a challenging task that involves different processes than in bulk-like structures. Besides the Schottky barrier height, the transfer length of electrons between metals and 2-D monolayers is a highly relevant parameter. For MoS2, both short (≤30 nm) and long (≥0.5 µm) values have been reported, corresponding to either an abrupt carrier injection at the contact edge or a more gradual transfer of electrons over a large contact area. Here we use ab initio quantum transport simulations to demonstrate that the presence of an oxide layer between a metallic contact and a MoS2 monolayer, for example, TiO2 in the case of titanium electrodes, favors an area-dependent process with a long transfer length, while a perfectly clean metal-semiconductor interface would lead to an edge process. These findings reconcile several theories that have been postulated about the physics of metal/MoS2 interfaces and provide a framework to design future devices with lower contact resistances.

Understanding the importance of autophagy in human diseases using Drosophila.

Autophagy is a lysosome-dependent intracellular degradation pathway that has been implicated in the pathogenesis of various human diseases, either positively or negatively impacting disease outcomes depending on the specific context. The majority of medical conditions including cancer, neurodegenerative diseases, infections and immune system disorders and inflammatory bowel disease could probably benefit from therapeutic modulation of the autophagy machinery. Drosophila represents an excellent model animal to study disease mechanisms thanks to its sophisticated genetic toolkit, and the conservation of human disease genes and autophagic processes. Here, we provide an overview of the various autophagy pathways observed both in flies and human cells (macroautophagy, microautophagy and chaperone-mediated autophagy), and discuss Drosophila models of the above-mentioned diseases where fly research has already helped to understand how defects in autophagy genes and pathways contribute to the relevant pathomechanisms.

Monitoring α-Synuclein Proteotoxicity in Drosophila Models.

Parkinson's disease is the second most common neurodegenerative disease without cure. It is characterized by α-synuclein accumulation and aggregation in dopaminergic and other types of neurons. Because α-synuclein accumulation leads to a toxic gain of function, its ectopic expression in Drosophila has been a useful in vivo model for testing modifiers of its toxicity. This chapter describes four assays: the rapid iterative negative geotaxis, rough eye phenotype, quantification of dopaminergic neuronal loss, and measurements of circadian effects.

Light from van der Waals quantum tunneling devices.

The understanding of and control over light emission from quantum tunneling has challenged researchers for more than four decades due to the intricate interplay of electrical and optical properties in atomic scale volumes. Here we introduce a device architecture that allows for the disentanglement of electronic and photonic pathways-van der Waals quantum tunneling devices. The electronic properties are defined by a stack of two-dimensional atomic crystals whereas the optical properties are controlled via an external photonic architecture. In van der Waals heterostructures made of gold, hexagonal boron nitride and graphene we find that inelastic tunneling results in the emission of photons and surface plasmon polaritons. By coupling these heterostructures to optical nanocube antennas we achieve resonant enhancement of the photon emission rate in narrow frequency bands by four orders of magnitude. Our results lead the way towards a new generation of nanophotonic devices that are driven by quantum tunneling.

Ubiquitylation Dynamics of the Clock Cell Proteome and TIMELESS during a Circadian Cycle.

Circadian clocks have evolved as time-measuring molecular devices to help organisms adapt their physiology to daily changes in light and temperature. Transcriptional oscillations account for a large fraction of rhythmic protein abundance. However, cycling of various posttranslational modifications, such as ubiquitylation, also contributes to shape the rhythmic protein landscape. In this study, we used an in vivo ubiquitin labeling assay to investigate the circadian ubiquitylated proteome of Drosophila melanogaster. We find that cyclic ubiquitylation affects MEGATOR (MTOR), a chromatin-associated nucleoporin that, in turn, feeds back to regulate the core molecular oscillator. Furthermore, we show that the ubiquitin ligase subunits CULLIN-3 (CUL-3) and SUPERNUMERARY LIMBS (SLMB) cooperate for ubiquitylating the TIMELESS protein. These findings stress the importance of ubiquitylation pathways in the Drosophila circadian clock and reveal a key component of this system.

Computationally efficient method for Fourier transform of highly chirped pulses for laser and parametric amplifier modeling.

We developed an improved approach to calculate the Fourier transform of signals with arbitrary large quadratic phase which can be efficiently implemented in numerical simulations utilizing Fast Fourier transform. The proposed algorithm significantly reduces the computational cost of Fourier transform of a highly chirped and stretched pulse by splitting it into two separate transforms of almost transform limited pulses, thereby reducing the required grid size roughly by a factor of the pulse stretching. The application of our improved Fourier transform algorithm in the split-step method for numerical modeling of CPA and OPCPA shows excellent agreement with standard algorithms.

Deubiquitinase Usp8 regulates α-synuclein clearance and modifies its toxicity in Lewy body disease.

In Parkinson's disease, misfolded α-synuclein accumulates, often in a ubiquitinated form, in neuronal inclusions termed Lewy bodies. An important outstanding question is whether ubiquitination in Lewy bodies is directly relevant to α-synuclein trafficking or turnover and Parkinson's pathogenesis. By comparative analysis in human postmortem brains, we found that ubiquitin immunoreactivity in Lewy bodies is largely due to K63-linked ubiquitin chains and markedly reduced in the substantia nigra compared with the neocortex. The ubiquitin staining in cells with Lewy bodies inversely correlated with the content and pathological localization of the deubiquitinase Usp8. Usp8 interacted and partly colocalized with α-synuclein in endosomal membranes and, both in cells and after purification, it deubiquitinated K63-linked chains on α-synuclein. Knockdown of Usp8 in the Drosophila eye reduced α-synuclein levels and α-synuclein-induced eye toxicity. Accordingly, in human cells, Usp8 knockdown increased the lysosomal degradation of α-synuclein. In the dopaminergic neurons of the Drosophila model, unlike knockdown of other deubiquitinases, Usp8 protected from α-synuclein-induced locomotor deficits and cell loss. These findings strongly suggest that removal of K63-linked ubiquitin chains on α-synuclein by Usp8 is a critical mechanism that reduces its lysosomal degradation in dopaminergic neurons and may contribute to α-synuclein accumulation in Lewy body disease.

Description of five dirofilariasis cases in South Hungary and review epidemiology of this disease for the country.

BACKGROUND: Dirofilariasis is an emerging zoonosis (supported by climate change) in Central Europe. Human infections are usually caused by Dirofilaria repens and Dirofilaria immitis with mediation of mosquito vectors. Aims of this publication were to report our dirofilariasis cases, and to summarize Hungarian epidemiological data by reviewing literature. METHODS AND RESULTS: We present five (four ophthalomological, one subcutaneous) cases observed within a 2-year period in Southern Hungary. Ages of infected patients were between 31 and 74 years. First case during pregnancy is also reported. There was no travel history in the anamnesis of patients which could explain acquisition of the infection. Moving, intact worms eliminated by surgical interventions were identified on the basis of morphological features as D. repens. Since the first report of human case, 115 further episodes (in addition to ours) were diagnosed in Hungary. Mean age of the patients was 47 years. Reviewing national reports, the ratio of subcutaneous infections was higher than that of the ocular ones (66 and 45, respectively). Evaluation of the territorial distribution of human episodes revealed that most infections occurred in patients living in the Danube-Tisza interfluvial region and eastern part of the country. However, sporadic cases were also found in western counties during 2014. CONCLUSION: Most of the Hungarian dirofilariasis cases were autochthonous infections. Occurrence in the western counties may suggest the spreading of this emerging zoonosis to these areas. Comprehensive monitoring and data analysis are desirable, therefore reporting the epidemiologic data in the case of human infections should be made mandatory.

Phase-sensitive amplification in a single bi-directional PPLN waveguide.

We investigate phase-sensitive amplification (PSA) and phase regeneration of a binary phase-shift keying (BPSK) signal using a single periodically poled lithium niobate (PPLN) waveguide. The PPLN is operated bi-directionally in order to simultaneously achieve phase correlated signals and phase-sensitive (PS) operation. We use injection-locking for carrier phase recovery and a lead zirconate titanate (PZT) fiber stretcher to correct path length deviations in the in-line phase regenerator. We observe a trade-off between high PS gain provided by high pumping power and stability of the device.

The CK2 kinase stabilizes CLOCK and represses its activity in the Drosophila circadian oscillator.

Phosphorylation is a pivotal regulatory mechanism for protein stability and activity in circadian clocks regardless of their evolutionary origin. It determines the speed and strength of molecular oscillations by acting on transcriptional activators and their repressors, which form negative feedback loops. In Drosophila, the CK2 kinase phosphorylates and destabilizes the PERIOD (PER) and TIMELESS (TIM) proteins, which inhibit CLOCK (CLK) transcriptional activity. Here we show that CK2 also targets the CLK activator directly. Downregulating the activity of the catalytic α subunit of CK2 induces CLK degradation, even in the absence of PER and TIM. Unexpectedly, the regulatory ß subunit of the CK2 holoenzyme is not required for the regulation of CLK stability. In addition, downregulation of CK2α activity decreases CLK phosphorylation and increases per and tim transcription. These results indicate that CK2 inhibits CLK degradation while reducing its activity. Since the CK1 kinase promotes CLK degradation, we suggest that CLK stability and transcriptional activity result from counteracting effects of CK1 and CK2.

Developmental-stage-specific regulation of the polyubiquitin receptors in Drosophila melanogaster.

Recognition of polyubiquitylated substrates by the proteasome is a highly regulated process that requires polyubiquitin receptors. We show here that the concentrations of the proteasomal and extraproteasomal polyubiquitin receptors change in a developmentally regulated fashion. The stoichiometry of the proteasomal p54/Rpn10 polyubiquitin receptor subunit, relative to that of other regulatory particle (RP) subunits falls suddenly at the end of embryogenesis, remains low throughout the larval stages, starts to increase again in the late third instar larvae and remains high in the pupae, adults and embryos. A similar developmentally regulated fluctuation was observed in the concentrations of the Rad23 and Dsk2 extraproteasomal polyubiquitin receptors. Depletion of the polyubiquitin receptors at the end of embryogenesis is due to the emergence of a developmentally regulated selective proteolytic activity. To follow the fate of subunit p54/Rpn10 in vivo, transgenic Drosophila melanogaster lines encoding the N-terminal half (NTH), the C-terminal half (CTH) or the full-length p54/Rpn10 subunit were established in the inducible Gal4-UAS system. The daughterless-Gal4-driven whole-body expression of the full-length subunit or its NTH did not produce any detectable phenotypic changes, and the transgenic products were incorporated into the 26S proteasome. The transgene-encoded CTH was not incorporated into the 26S proteasome, caused third instar larval lethality and was found to be multi-ubiquitylated. This modification, however, did not appear to be a degradation signal because the half-life of the CTH was over 48 hours. Accumulation of the CTH disturbed the developmentally regulated changes in subunit composition of the RP and the emergence of the selective proteolytic activity responsible for the depletion of the polyubiquitin receptors. Build-up of subunit p54/Rpn10 in the RP had already started in 84-hour-old larvae and reached the full complement characteristic of the non-larval developmental stages at the middle of the third instar larval stage, just before these larvae perished. Similar shifts were observed in the concentrations of the Rad23 and Dsk2 polyubiquitin receptors. The postsynthetic modification of CTH might be essential for this developmental regulation, or it might regulate an essential extraproteasomal function(s) of subunit p54/Rpn10 that is disturbed by the expression of an excess of CTH.