Double-clad ytterbium-doped tapered fiber with circular birefringence as a gain medium for structured light.

Amplifying radially and azimuthally polarized beams is a significant challenge due to the instability of the complex beam shape and polarization in inhomogeneous environment. In this Letter, we demonstrated experimentally an efficient approach to directly amplify cylindrical-vector beams with axially symmetric polarization and doughnut-shaped intensity profile in a picosecond MOPA system based on a double-clad ytterbium-doped tapered fiber. To prevent polarization and beam shape distortion during amplification, for the first time to the best of our knowledge, we proposed using the spun architecture of the tapered fiber. In contrast to an isotropic fiber architecture, a spun configuration possessing nearly circular polarization eigenstates supports stable wavefront propagation. Applying this technique, we amplified the cylindrical-vector beam with 10 ps pulses up to 22 W of the average power at a central wavelength of 1030 nm and a repetition rate of 15 MHz, maintaining both mode and polarization stability.

Active tapered double-clad fiber with low birefringence.

We addressed the problem of a state of polarization (SOP) drift caused by heating under intense clad pumping in different types of active tapered double-clad fibers. We investigated experimentally the variations of the SOP and degree of polarization (DOP) under clad pumping in polarization-maintaining (PANDA type) and regular (non-PM) Yb-doped double-clad large mode area tapered fibers. We discovered that the birefringence of active fibers is highly dependent on the launched pump power. To solve the problem of the SOP drift in active large mode are fibers, we, for the first time to the best of our knowledge, presented an active double-clad fiber with low intrinsic birefringence as a gain medium. An Yb-doped spun tapered double-clad fiber (sT-DCF) with intrinsic birefringence as low as 1.45×10-8 was manufactured and experimentally studied. We have proved experimentally that the DOP and SOP remains more stable in sT-DCF with increasing pump power compared to PM PANDA-type and regular non-PM tapered double-clad fibers. In particular, the SOP drift in sT-DCF is almost one order of magnitude less than in other tapered fibers, while the DOP drift in sT-DCF is comparable with the drift in PANDA-type fiber and one order of magnitude less than in the non-PM tapered fiber. An active sT-DCF showing efficient amplification was demonstrated in an all-fiber-based picosecond master-oscillator power-amplifier scheme. The system delivered 50 ps pulses at 1040 nm with an average power of 50 W, 34 dB gain, 26 µm MFD and perfect beam quality.

Single-frequency 100†ns / 0.5 mJ laser pulses from all-fiber double clad ytterbium doped tapered fiber amplifier.

High peak power, narrow linewidth sources continue to be in high demand. Fiber amplifiers are a compelling option to scale peak power of long 100-ns-pulses because of their compact size and robustness. Unfortunately, stimulated Brillouin scattering (SBS) limits peak power of narrow linewidth fiber sources causing instability. We demonstrate SBS suppression for 130-ns pulses from a 5 MHz linewidth seed laser in a fiber amplifier by using tapered fiber with large 50 µm diameter core in the output. The longitudinal change in the core diameter induces frequency shift in the SBS gain peak and the back-travelling Stokes wave is suppressed towards smaller core. We reach 2.2 kW peak power with 18.7 dB polarization extinction ratio and record breaking 4 kW peak power by exciting both polarization states of the polarization maintaining tapered fiber. The output beam quality equals to single mode fibers with M2 = 1.08.

IRE1α inhibition decreased TXNIP/NLRP3 inflammasome activation through miR-17-5p after neonatal hypoxic-ischemic brain injury in rats.

BACKGROUND: The endoplasmic reticulum (ER) is responsible for the control of correct protein folding and protein function which is crucial for cell survival. However, under pathological conditions, such as hypoxia-ischemia (HI), there is an accumulation of unfolded proteins thereby triggering the unfolded protein response (UPR) and causing ER stress which is associated with activation of several stress sensor signaling pathways, one of them being the inositol requiring enzyme-1 alpha (IRE1α) signaling pathway. The UPR is regarded as a potential contributor to neuronal cell death and inflammation after HI. In the present study, we sought to investigate whether microRNA-17 (miR-17), a potential IRE1α ribonuclease (RNase) substrate, arbitrates downregulation of thioredoxin-interacting protein (TXNIP) and consequent NLRP3 inflammasome activation in the immature brain after HI injury and whether inhibition of IRE1α may attenuate inflammation via miR-17/TXNIP regulation. METHODS: Postnatal day 10 rat pups (n = 287) were subjected to unilateral carotid artery ligation followed by 2.5 h of hypoxia (8% O2). STF-083010, an IRE1α RNase inhibitor, was intranasally delivered at 1 h post-HI or followed by an additional one administration per day for 2 days. MiR-17-5p mimic or anti-miR-17-5p inhibitor was injected intracerebroventricularly at 48 h before HI. Infarct volume and body weight were used to evaluate the short-term effects while brain weight, gross and microscopic brain tissue morphologies, and neurobehavioral tests were conducted for the long-term evaluation. Western blots, immunofluorescence staining, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), and co-immunoprecipitation (Co-IP) were used for mechanism studies. RESULTS: Endogenous phosphorylated IRE1α expression was significantly increased after HI. Intranasal administration of STF-083010 alleviated brain injury and improved neurological behavior. MiR-17-5p expression was reduced after HI, and this decrease was attenuated by STF-083010 treatment. MiR-17-5p mimic administration ameliorated TXNIP expression, NLRP3 inflammasome activation, caspase-1 cleavage, and IL-1ß production, as well as brain infarct volume. Conversely, anti-miR-17-5p inhibitor reversed IRE1α inhibition-induced decrease in TXNIP expression and inflammasome activation, as well as exacerbated brain injury after HI. CONCLUSIONS: IRE1a-induced UPR pathway may contribute to inflammatory activation and brain injury following neonatal HI. IRE1a activation, through decay of miR-17-5p, elevated TXNIP expression to activate NLRP3 inflammasome and aggravated brain damage.

Ultra-large core birefringent Yb-doped tapered double clad fiber for high power amplifiers.

We present a birefringent Yb-doped tapered double-clad fiber with a record core diameter of 96 µm. An impressive gain of over 38 dB was demonstrated for linearly polarized CW and pulsed sources at a wavelength of 1040 nm. For the CW regime the output power was70 W. For a mode-locked fiber laser a pulse energy of 28 µJ with 292 kW peak power was reached at an average output power of 28 W for a 1 MHz repetition rate. The tapered double-clad fiber has a high value of polarization extinction ratio at 30 dB and is capable of delivering the linearly polarized diffraction-limited beam (M2 = 1.09).

Human papillomavirus type 16 E6* induces oxidative stress and DNA damage.

UNLABELLED: High-risk types of human papillomavirus (HPV) are the causative agents of virtually all cases of cervical cancer and a significant proportion of other anogenital cancers, as well as both oral and pharyngeal cancers. The high-risk types encode two viral oncogenes, E6 and E7, which work together to initiate cell transformation. Multiple steps involving the activities and interactions of both viral and cellular proteins are involved in the progression from HPV infection to cell transformation to cancer. The E6 oncoprotein is expressed as several isoforms: a full-length variant referred to as E6 and a few shorter isoforms collectively referred to as E6*. In this study, we found that expression of E6* increased the level of reactive oxygen species (ROS) in both HPV-positive and HPV-negative cells. This increased oxidative stress led to higher levels of DNA damage, as assessed by the comet assay, quantification of 8-oxoguanine, and poly(ADP-ribose) polymerase 1. The observed increase in ROS may be due to a decrease in cellular antioxidant activity, as we found that E6* expression also led to decreased expression of superoxide dismutase isoform 2 and glutathione peroxidase. These studies indicate that E6* may play an important role in virus-induced mutagenesis by increasing oxidative stress and DNA damage. IMPORTANCE: Our findings demonstrate for the first time that an HPV gene product, E6*, can increase ROS levels in host cells. This ability may play a significant role both in the viral life cycle and in cancer development, because an increase in oxidative DNA damage may both facilitate HPV genome amplification and increase the probability of HPV16 DNA integration. Integration, in turn, is thought to be an important step in HPV-mediated carcinogenesis.

Docosahexanoic acid antagonizes TNF-α-induced necroptosis by attenuating oxidative stress, ceramide production, lysosomal dysfunction, and autophagic features.

OBJECTIVE: It was previously reported that docosahexanoic acid (DHA) reduces TNF-α-induced necrosis in L929 cells. However, the mechanisms underlying this reduction have not been investigated. The present study was designed to investigate cellular and biochemical mechanisms underlying the attenuation of TNF-α-induced necroptosis by DHA in L929 cells. METHODS: L929 cells were pre-treated with DHA prior to exposure to TNF-α, zVAD, or Necrostatin-1 (Nec-1). Cell death and survival were assessed by MTT and caspase activity assays, and microscopic visualization. Reactive oxygen species (ROS) were measured by flow cytometry. C16- and C18-ceramides were measured by mass spectrometry. Lysosomal membrane permeabilization (LMP) was evaluated by fluorescence microscopy and flow cytometry using Acridine Orange. Cathepsin L activation was evaluated by immunoblotting and fluorescence microscopy. Autophagy was assessed by immunoblotting of LC3-II and Beclin. RESULTS: Exposure of L929 cells to TNF-α alone for 24 h induced necroptosis, as evidenced by the inhibition of cell death by Nec-1, absence of caspase-3 activity and Lamin B cleavage, and morphological analysis. DHA attenuated multiple biochemical events associated with TNF-α-induced necroptosis, including ROS generation, ceramide production, lysosomal dysfunction, cathepsin L activation, and autophagic features. DHA also attenuated zVAD-induced necroptosis but did not attenuate the enhanced apoptosis and necrosis induced by the combination of TNF-α with Actinomycin D or zVAD, respectively, suggesting that its protective effects might be limited by the strength of the cell death insult induced by TNF-α. CONCLUSIONS: DHA effectively attenuates TNF-α-induced necroptosis and autophagy, most likely via its ability to inhibit TNF-α-induced sphingolipid metabolism and oxidative stress. These results highlight the role of this Omega-3 fatty acid in antagonizing inflammatory cell death.

DNA damage induces down-regulation of UDP-glucose ceramide glucosyltransferase, increases ceramide levels and triggers apoptosis in p53-deficient cancer cells.

DNA damaging agents typically induce an apoptotic cascade in which p53 plays a central role. However, absence of a p53-mediated response does not necessarily abrogate programmed cell death, due to the existence of p53-independent apoptotic pathways, such as those mediated by the pro-apoptotic molecule ceramide. We compared ceramide levels before and after DNA damage in human osteosarcoma (U2OS) and colon cancer (HCT116) cells that were either expressing or deficient in p53. When treated with mitomycin C, p53-deficient cells, but not p53-expressing cells, showed a marked increase in ceramide levels. Microarray analysis of genes involved in ceramide metabolism identified acid ceramidase (ASAH1, up-regulated), ceramide glucosyltransferase (UGCG, down-regulated), and galactosylceramidase (GALC, up-regulated) as the three genes most affected. Experiments employing pharmacological and siRNA agents revealed that inhibition of UGCG is sufficient to increase ceramide levels and induce cell death. When inhibition of UGCG and treatment with mitomycin C were combined, p53-deficient, but not p53-expressing cells, showed a significant increase in cell death, suggesting that the regulation of sphingolipid metabolism could be used to sensitize cells to chemotherapeutic drugs.

Mass spectrometric studies on epigenetic interaction networks in cell differentiation.

Arrest of cell differentiation is one of the leading causes of leukemia and other cancers. Induction of cell differentiation using pharmaceutical agents has been clinically attempted for the treatment of these cancers. Epigenetic regulation may be one of the underlying molecular mechanisms controlling cell proliferation or differentiation. Here, we report on the use of proteomics-based differential protein expression analysis in conjunction with quantification of histone modifications to decipher the interconnections among epigenetic modifications, their modifying enzymes or mediators, and changes in the associated pathways/networks that occur during cell differentiation. During phorbol-12-myristate 13-acetate-induced differentiation of U937 cells, fatty acid synthesis and its metabolic processing, the clathrin-coated pit endocytosis pathway, and the ubiquitin/26 S proteasome degradation pathways were up-regulated. In addition, global histone H3/H4 acetylation and H2B ubiquitination were down-regulated concomitantly with impaired chromatin remodeling machinery, RNA polymerase II complexes, and DNA replication. Differential protein expression analysis established the networks linking histone hypoacetylation to the down-regulated expression/activity of p300 and linking histone H2B ubiquitination to the RNA polymerase II-associated FACT-RTF1-PAF1 complex. Collectively, our approach has provided an unprecedentedly systemic set of insights into the role of epigenetic regulation in leukemia cell differentiation.

Mode evolution in long tapered fibers with high tapering ratio.

We have experimentally studied fundamental mode propagation in few meters long, adiabatically tapered step-index fibers with high numerical aperture, core diameter up to 117 µm (V = 38) and tapering ratio up to 18. The single fundamental mode propagation was confirmed by several techniques that reveal no signature of higher-order mode excitation. It can be, therefore, concluded that adiabatic tapering is a powerful method for selective excitation of the fundamental mode in highly multimode large-mode-area fibers. Annular near field distortion observed for large output core diameters was attributed to built-in stress due to thermal expansion mismatch between core and cladding materials. The mechanical stress could be avoided by an appropriate technique of fiber preform fabrication and drawing, which would prevent the mode field deformation and lead to reliable diffraction-limited fundamental mode guiding for very large core diameters.

Thermo-optic coefficients study in KGd(WO4)2 and KY(WO4)2 by a modified minimum deviation method.

Thermo-optic coefficients anisotropy was characterized in monoclinic potassium (rare-earth) double tungstates KGd(WO4)2 and KY(WO4)2 in the spectral range of 0.44-0.63 µm by a modified minimum deviation method. This approach takes into account the changes in the shape and dimensions of the prismatic sample caused by the anisotropic thermal expansion effect under uniform heating together with the conventional measurements of minimum deviation angle. At room temperature at the wavelength of 633 nm, principal refractive indices for KGdW are n(p)=2.0135, n(m)=2.0458, and n(g)=2.0860, and for KYW they are n(p)=1.9979, n(m)=2.0396, and n(g)=2.0869. Optical axes position for KGdW (KYW) crystals is N(g) ± 42.60 (N(g) ± 44.1°) in the N(p)-N(g) plane. For both crystals, all the thermo-optic coefficients are negative and equal dn(p)/dT=-10.6, dn(m)/dT=-8.4, and dn(g)/dT=-15.2 for KGdW, and dn(p)/dT=-10.1, dn(m)/dT=-7.3, and dn(g)/dT=-8.4 for KYW [10(-6) K(-1)] (at 633 nm).

Principles and performance of tapered fiber lasers: from uniform to flared geometry.

We have studied the recently demonstrated concept of fiber lasers based on active tapered double-clad fiber (T-DCF) in copropagating and counterpropagating configurations, both theoretically and experimentally, and compared the performance to fiber lasers based on conventional cylindrical fibers in end-pumped configurations. Specific properties of T-DCFs were considered theoretically using a rate-equation model developed for tapered fibers, and a detailed comparative study was carried out experimentally. Furthermore, we have studied mode coupling effects in long adiabatic tapers due to coiling and local bending. The results allow us to conclude that, with proper fiber design, the T-DCF technology offers a high-potential alternative for bright, cost-effective fiber devices.

Proton-induced DNA damage promotes integration of foreign plasmid DNA into human genome.

High-risk human papillomaviruses (HPVs) cause virtually all cervical cancer cases and are also associated with other types of anogenital and oropharyngeal cancers. Normally, HPV exists as a circular episomal DNA in the infected cell. However, in some instances, it integrates into the human genome in such a way as to enable increased expression of viral oncogenes, thereby leading to carcinogenesis. Since viral integration requires breaks in both viral and human genomes, DNA damage likely plays a key role in this critical process. One potentially significant source of DNA damage is exposure to elevated doses of ionizing radiation. Natural background radiation is ubiquitous; however, some populations, including radiological workers, radiotherapy patients, and astronauts, are exposed to significantly higher radiation doses, as well as to different types of radiation such as particle radiation. We hypothesize that ionizing radiation-induced DNA damage facilitates the integration of HPV into the human genome, increasing the risk of developing HPV-related cancers in the exposed population. To test this, we first determined the kinetics of DNA damage in keratinocytes exposed to ionizing radiation (protons) by assessing γ-H2AX foci formation using immunofluorescence (direct damage), and also measured ROS and 8-oxoG levels via DCFDA and Avidin-FITC (indirect damage).As anticipated, direct DNA damage was observed promptly, within 30 min, whereas indirect DNA damage was delayed due to the time required for ROS to accumulate and cause oxidative damage. Although radiation was lethal at high doses, we were able to establish an experimental system where radiation exposure (protons and X-rays) induced DNA damage dose-dependently without causing major cytotoxic effects as assessed by several cytotoxicity assays. Most importantly, we explored the impact of radiation exposure on integration frequency using a clonogenic assay and demonstrated that as predicted, proton-induced DNA damage promotes the integration of HPV-like foreign DNA in oral keratinocytes. Overall, the insights gained from this work enable us to better understand the contribution of radiation exposure and DNA damage to HPV-mediated carcinogenesis and direct us toward strategies aimed at preventing malignancies in HPV-infected individuals.

Splice variants of mda-7/IL-24 differentially affect survival and induce apoptosis in U2OS cells.

Interleukin-24 (mda-7/IL-24) is a cytokine in the IL-10 family that has received a great deal of attention for its properties as a tumor suppressor and as a potential treatment for cancer. In this study, we have identified and characterized five alternatively spliced isoforms of this gene. Several, but not all of these isoforms induce apoptosis in the osteosarcoma cell line U2OS, while none affect the survival of the non-cancerous NOK cell line. One of these isoforms, lacking three exons and encoding the N-terminal end of the mda-7/IL-24 protein sequence, caused levels of apoptosis that were higher than those caused by the full-length mda-7/IL-24 variant. Additionally, we found that the ratio of isoform expression can be modified by the splice factor SRp55. This regulation suggests that alternative splicing of mda-7/IL-24 is under tight control in the cell, and can be modified under various cellular conditions, such as DNA damage. In addition to providing new insights into the function of an important tumor suppressor gene, these findings may also point toward new avenues for cancer treatment.

Complexes of human papillomavirus type 16 E6 proteins form pseudo-death-inducing signaling complex structures during tumor necrosis factor-mediated apoptosis.

High-risk strains of human papillomavirus (HPV) such as HPV type 16 (HPV16) and HPV18 are causative agents of most human cervical carcinomas. E6, one of the oncogenes encoded by HPV16, possesses a number of biological and transforming functions. We have previously shown that the binding of E6 to host apoptotic proteins such as tumor necrosis factor (TNF) R1, the adaptor protein FADD, and procaspase 8 results in a significant modification of the normal flow of apoptotic events. For example, E6 can bind to and accelerate the degradation of FADD. In addition, full-length E6 binds to the TNF R1 death domain and can also bind to and accelerate the degradation of procaspase 8. In contrast, the binding of small splice isoforms known as E6* results in the stabilization of procaspase 8. In this report, we propose a model for the ability of HPV16 E6 to both sensitize and protect cells from TNF as well as to protect cells from Fas. We demonstrate that both the level of E6 expression and the ratio between full-length E6 and E6* are important factors in the modification of the host extrinsic apoptotic pathways and show that at high levels of E6 expression, the further sensitization of U2OS, NOK, and Ca Ski cells to TNF-mediated apoptosis is most likely due to the formation of a pseudo-death-inducing signaling complex structure that includes complexes of E6 proteins.

Highly efficient 750 W tapered double-clad ytterbium fiber laser.

The results of theoretical and experimental studies of active tapered double-clad fibers, intending the optimization of its imperative parameters--tapering ratio, longitudinal profile, core/cladding diameters ratio, are presented. Using a refined taper geometry we have demonstrated power scaling of a ytterbium fiber laser pumped by low-brightness, cost-effective laser diodes up to 750 W, with 80% efficiency.

Actively Q-switched 1.6-mJ tapered double-clad ytterbium-doped fiber laser.

We have demonstrated an actively Q-switched tapered double-clad fiber laser capable of single-shot generation of 1.6-mJ, 64-ns pulses. The active medium based on tapered double-clad fiber is shown to exhibit a reduced level of amplified spontaneous emission which allows for high-energy pulse extraction at extremely low repetition rates.

Picosecond Yb-doped tapered fiber laser system with 1.26 MW peak power and 200 W average output power.

We demonstrate a compact picosecond master-oscillator power-amplifier (MOPA) system based on an Yb-doped polarization-maintaining double-clad tapered fiber (T-DCF) delivering pulses with over 1.26 MW peak power and average output power up to 200 W preserving near diffraction limited beam quality. The unique properties of an active tapered fiber enable to amplify the seed pulses directly with no need for applying of additional stretching technique. This simplified laser system can find the practical implementation in industrial micromachining.

Methylomes in Vegans versus Pescatarians and Nonvegetarians.

Epigenetic studies in animal models have demonstrated that diet affects gene regulation by altering methylation patterns. We interrogated methylomes in humans who have different sources of protein in their diet. We compared methylation of DNA isolated from buffy coat in 38 vegans, 41 pescatarians and 68 nonvegetarians. Methylation data were obtained using Infinium HumanMethylation450 arrays and analyzed using the Partek Genomic software. Differences in differentially methylated sites were small, though with the use of relaxed statistical tests we did identify diet-associated differences. To further test the validity of these observations, we performed separate and independent comparisons of the methylation differences between vegans and nonvegetarians, and between vegans and pescatarians. The detected differences were then examined to determine if they were enriched in specific pathways. Pathway analysis revealed enrichment of several specific processes, including homeobox transcription and glutamate transport. The detected differences in DNA methylation patterns between vegans, pescatarians, and nonvegetarians enabled us to identify 77 CpG sites that may be sensitive to diet and/or lifestyle, though high levels of individual-specific differences were also noted.

DNA Methylation Profiles of Vegans and Non-Vegetarians in the Adventist Health Study-2 Cohort.

We sought to determine if DNA methylation patterns differed between vegans and non-vegetarians in the Adventist Health Study-2 cohort. Genome-wide DNA methylation derived from buffy coat was profiled in 62 vegans and 142 non-vegetarians. Using linear regression, methylation of CpG sites and genes was categorized or summarized according to various genic/intergenic regions and CpG island-related regions, as well as the promoter. Methylation of genes was measured as the average methylation of available CpG's annotated to the nominated region of the respective gene. A permutation method defining the null distribution adapted from Storey et al. was used to adjust for false discovery. Differences in methylation of several CpG sites and genes were detected at a false discovery rate < 0.05 in region-specific and overall analyses. A vegan diet was associated predominantly with hypomethylation of genes, most notably methyltransferase-like 1 (METTL1). Although a limited number of differentially methylated features were detected in the current study, the false discovery method revealed that a much larger proportion of differentially methylated genes and sites exist, and could be detected with a larger sample size. Our findings suggest modest differences in DNA methylation in vegans and non-vegetarians, with a much greater number of detectable significant differences expected with a larger sample.