Dual-output kilohertz pump laser for high-energy picosecond OPCPA.

A dual-output thin-disk picosecond laser operating at 100 W with 1 kHz repetition rate is reported in this Letter. By electronically adjusting the amplitude of the optical seed pulses that are injected into the laser cavity, the energy extracted from the gain medium can be shared between two pulses. Amplified double pulses are subsequently spatially separated into two independent beams by a fast Pockels cell, compressed in one common compressor, and frequency-doubled with ∼70% efficiency. This approach significantly decreases strain on the optics, as well as nonlinear effects, and is advantageous for power scaling.

120 mJ, 1 kHz, picosecond laser at 515 nm.

We report on a 1 kHz, 515 nm laser system, based on a commercially available 230 W average power Yb:YAG thin-disk regenerative amplifier, developed for pumping one of the last optical parametric chirped pulse amplification (OPCPA) stages of the Allegra laser system at ELI Beamlines. To avoid problems with self-focusing of picosecond pulses, the 1030 nm output pulses are compressed and frequency doubled with an LBO crystal in vacuum. Additionally, development of a thermal management system was needed to ensure stable phase matching conditions at high average power. The resulting 515 nm pulses have an energy of more than 120 mJ with SHG efficiency of 60% and an average RMS stability of 1.1% for more than 8 h.

Mitigation of laser-induced contamination in vacuum in high-repetition-rate high-peak-power laser systems.

Vacuum chambers are frequently used in high-energy, high-peak-power laser systems to prevent deleterious nonlinear effects, which can result from propagation in air. In the vacuum sections of the Allegra laser system at ELI-Beamlines, we observed degradation of several optical elements due to laser-induced contamination (LIC). This contamination is present on surfaces with laser intensity above 30GW/cm2 with wavelengths of 515, 800, and 1030 nm. It can lead to undesired absorption on diffraction gratings, mirrors, and crystals and ultimately to degradation of the laser beam profile. Because the Allegra laser is intended to be a high-uptime source for users, such progressive degradation is unacceptable for operation. Here, we evaluate three methods of removing LIC from optics in vacuum. One of them, the radio-frequency-generated plasma cleaning, appears to be a suitable solution from the perspective of operating a reliable, on-demand source for users.

Bordetellapertussis Acetylome is Shaped by Lysine Deacetylase Bkd1.

Post-translational modifications of proteins enable swift physiological adaptation of cells to altered growth conditions and stress. Aside from protein phosphorylation, acetylation on ε-amino groups of lysine residues (N-ε-lysine acetylation) represents another important post-translational modification of proteins. For many bacterial pathogens, including the whooping cough agent Bordetella pertussis, the role and extent of protein acetylation remain to be defined. We expressed in Escherichia coli the BP0960 and BP3063 genes encoding two putative deacetylases of B. pertussis and show that BP0960 encodes a lysine deacetylase enzyme, named Bkd1, that regulates acetylation of a range of B. pertussis proteins. Comparison of the proteome and acetylome of a Δbkd1 mutant with the proteome and acetylome of wild-type B. pertussis (PRIDE ID. PXD016384) revealed that acetylation on lysine residues may modulate activities or stabilities of proteins involved in bacterial metabolism and histone-like proteins. However, increased acetylation of the BvgA response regulator protein of the B. pertussis master virulence-regulating BvgAS two-component system affected neither the total levels of produced BvgA nor its phosphorylation status. Indeed, the Δbkd1 mutant was not impaired in the production of key virulence factors and its survival within human macrophages in vitro was not affected. The Δbkd1 mutant exhibited an increased growth rate under carbon source-limiting conditions and its virulence in the in vivo mouse lung infection model was somewhat affected. These results indicate that the lysine deacetylase Bkd1 and N-ε-lysine acetylation primarily modulate the general metabolism rather than the virulence of B. pertussis.

Stability mechanism of picosecond supercontinuum in YAG.

Stable picosecond supercontinuum generated in long crystals is an excellent means of seeding broadband, high-energy CPA systems. The generated output energy and spectrum can be almost three times as stable as the pump for a wide range of input pulse parameters. In this work, we show this is an intrinsic property for crystals longer than the filament and for a range of input energy values. We present a description of the stability mechanism in both the visible and infrared regions together with experimental data that support the theoretical explanation.

Picosecond pulse generated supercontinuum as a stable seed for OPCPA.

We present a stable supercontinuum (SC) generated in a bulk YAG crystal, pumped by 3 ps chirped pulses at 1030 nm. The SC is generated in a loose focus geometry in a 13 cm long YAG crystal, allowing for stable and robust single-filament generation. The SC energy stability exceeds that of the pump laser by almost a factor of 3. Additionally, we show that the SC spectrum has long-term stability and that the SC is coherent and compressible by compressing the portions of SC spectra close to the corresponding Fourier limit. This makes the picosecond-pulse-driven SC a suitable stable seed for OPCPA amplifiers.

GLD-4-mediated translational activation regulates the size of the proliferative germ cell pool in the adult C. elegans germ line.

To avoid organ dysfunction as a consequence of tissue diminution or tumorous growth, a tight balance between cell proliferation and differentiation is maintained in metazoans. However, cell-intrinsic gene expression mechanisms controlling adult tissue homeostasis remain poorly understood. By focusing on the adult Caenorhabditis elegans reproductive tissue, we show that translational activation of mRNAs is a fundamental mechanism to maintain tissue homeostasis. Our genetic experiments identified the Trf4/5-type cytoplasmic poly(A) polymerase (cytoPAP) GLD-4 and its enzymatic activator GLS-1 to perform a dual role in regulating the size of the proliferative zone. Consistent with a ubiquitous expression of GLD-4 cytoPAP in proliferative germ cells, its genetic activity is required to maintain a robust proliferative adult germ cell pool, presumably by regulating many mRNA targets encoding proliferation-promoting factors. Based on translational reporters and endogenous protein expression analyses, we found that gld-4 activity promotes GLP-1/Notch receptor expression, an essential factor of continued germ cell proliferation. RNA-protein interaction assays documented also a physical association of the GLD-4/GLS-1 cytoPAP complex with glp-1 mRNA, and ribosomal fractionation studies established that GLD-4 cytoPAP activity facilitates translational efficiency of glp-1 mRNA. Moreover, we found that in proliferative cells the differentiation-promoting factor, GLD-2 cytoPAP, is translationally repressed by the stem cell factor and PUF-type RNA-binding protein, FBF. This suggests that cytoPAP-mediated translational activation of proliferation-promoting factors, paired with PUF-mediated translational repression of differentiation factors, forms a translational control circuit that expands the proliferative germ cell pool. Our additional genetic experiments uncovered that the GLD-4/GLS-1 cytoPAP complex promotes also differentiation, forming a redundant translational circuit with GLD-2 cytoPAP and the translational repressor GLD-1 to restrict proliferation. Together with previous findings, our combined data reveals two interconnected translational activation/repression circuitries of broadly conserved RNA regulators that maintain the balance between adult germ cell proliferation and differentiation.

Broadband OPCPA system with 11 mJ output at 1 kHz, compressible to 12 fs.

We report on a broadband OPCPA system, pumped at 515 nm by frequency doubled Yb:YAG thin disk lasers. The system delivers 11.3 mJ pulses at a central wavelength of 800 nm with a spatial beam quality of M2 = 1.25 and > 25% pump-to-signal conversion efficiency. The broadband pulses were demonstrated to be compressible to 12 fs using a chirped mirror compressor.

Thin disk amplifier-based 40 mJ, 1 kHz, picosecond laser at 515 nm.

We report on a frequency-doubled picosecond Yb:YAG thin disk regenerative amplifier, developed as a pump laser for a kilohertz repetition rate OPCPA. At a repetition rate of 1 kHz, the compressed output of the regenerative amplifier has a pulse duration of 1.2 ps and pulse energy of 90 mJ with energy stability of σ < 0.8% and M2 < 1.2. The pulses are frequency doubled in an LBO crystal yielding 42 mJ at 515 nm.

Impact of meteorological and environmental factors on the spatial distribution of Fasciola hepatica in beef cattle herds in Sweden.

BACKGROUND: Fasciola hepatica is a parasite with a significant impact on ruminant livestock production. Previous studies in north-west Europe have described its geographical distribution and determined potential predictors of fasciolosis using geographical information system (GIS) and regression modelling. In Sweden, however, information about the distribution of fasciolosis is limited. This study examined the geographical distribution of F. hepatica and identified high-risk areas for beef cattle in Sweden and sought to characterise potential predictors. Beef cattle serum samples were collected during winter 2006-2007 from 2135 herds which were examined for F. hepatica antibodies by enzyme-linked immunosorbent assay (ELISA). Fasciolosis distribution maps were created using GIS based on postcode location of seropositive herds. Spatial scan analysis (SaTScan) was performed to determine high-risk areas. Using datasets on animal density, temperature, precipitation and Corine land cover data, including soil type and soil mineral concentrations in Sweden, bivariate and multiple logistic regression analyses were carried out in R software to reveal potential predictors of F. hepatica infection. RESULTS: Overall herd seroprevalence of F. hepatica in beef cattle was 9.8 % (95 % CI: 8.6-11.1). An irregular spatial distribution of F. hepatica, with two main clusters, was observed in south-west Sweden. The most northerly occurrence of F. hepatica in the world was documented. The final model explained 15.8 % of the variation in F. hepatica distribution in study herds. Absence of coniferous forest was the variable with the highest predictive value. Precipitation in July-September, Dystric Cambisol, Dystric Regosol, and P and Cu concentrations in soil were other negative predictors. Beef cattle herd density, Dystric Leptosol and Fe concentration were positive predictors. CONCLUSIONS: The spatial distribution of F. hepatica in Swedish beef cattle herds is influenced by multi-factorial effects. Interestingly, absence of coniferous forest, herd density, specific soil type and concentration of some soil minerals are more important predictors than climate factors.

Pulse synchronization system for picosecond pulse-pumped OPCPA with femtosecond-level relative timing jitter.

A simple and compact scheme for synchronization of the pump and signal pulses for short-pulse OPCPA is demonstrated. Relative timing jitter of 17 fs RMS is achieved (1% of the pump pulse duration) and the system remains locked for hours. The scheme uses a balanced optical cross correlator to detect relative delays between the pump and signal pulses and can be operated with just 10's of µJ of pump energy and pJ-level signal energies.

Anorectal dysfunction in multiple sclerosis patients: A pilot study on the effect of an individualized rehabilitation approach.

BACKGROUND: Anorectal dysfunction (ARD), especially bowel incontinence, frequently compromises the quality of life in multiple sclerosis (MS) patients. The effect of rehabilitation procedures has not been clearly established. OBJECTIVE: To determine the effect of an individualized rehabilitation approach on bowel incontinence and anorectal pressures. METHODS: MS patients with ARD underwent 6-months of individually targeted biofeedback rehabilitation. High resolution anorectal manometry (HRAM) and St. Mark's Fecal Incontinence Scores (SMIS) were completed prior to rehabilitation, after 10 weeks of supervised physiotherapy, and after 3 months of self-treatment. RESULTS: Ten patients (50%) completed the study. Repeated measures analysis of variance (ANOVA) demonstrated significant improvement in the SMIS questionnaire over time [14.00 baseline vs. 9.70 after supervised physiotherapy vs. 9.30 after self-treatment (p = 0.005)]. No significant improvements over time were noted in any HRAM readings: maximal pressure [49.85 mmHg baseline vs. 57.60 after supervised physiotherapy vs. 60.88 after self-treatment (p = 0.58)], pressure endurance [36.41 vs. 46.89 vs. 49.95 (p = 0.53)], resting pressure [55.83, vs 52.69 vs. 51.84 (p = 0.704)], or area under the curve [230.0 vs. 520.8 vs. 501.9 (p = 0.16)]. CONCLUSIONS: The proposed individualized rehabilitation program supports a positive overall effect on anorectal dysfunction in MS patients.

Abdominal wall tension increases using Dynamic Neuromuscular Stabilization principles in different postural positions.

BACKGROUND: Intra-abdominal pressure (IAP) is an important mechanism stabilizing the spine and trunk. IAP regulation depends on the coordination of abdominal muscles, diaphragm and pelvic floor muscles. OBJECTIVE: To determine the differences in abdominal wall tension (AWT) of various postural positions, first without any correction, then after verbal and manual instructions according to Dynamic Neuromuscular Stabilization (DNS) principles. METHODS: In a cross-sectional observational study, thirty healthy individuals (mean age = 22.73 ± 1.91 years) were fitted with two Ohmbelt sensors contralaterally above the inguinal ligament and in the upper lumbar triangle. AWT was measured during five postural positions: sitting, supine with legs raised, squat, bear and hang position. First, spontaneous AWT was measured, then again after manual and verbal instructions following DNS principles. RESULTS: AWT increased significantly with DNS instructions compared to spontaneous activation. Both sensors recorded significant increases (p < .01; Cohen's d = -1.13 to -2.06) in all observed postural situations. The increase in activity occurred simultaneously on both sensors, with no significant differences noted in pressure increases between the sensors. The greatest activation for both sensors occurred in the bear position. Significant increases in activity were identified for both sensors in the supine leg raise position and in the bear position compared to spontaneous activation in sitting (p < .001). There were no statistically significant differences (for both sensors) between women and men in any position. CONCLUSION: The amount of AWT significantly increases after verbal and manual instructions according to DNS. The greatest abdominal wall activation was achieved in the bear position.

Dazap2 modulates transcription driven by the Wnt effector TCF-4.

A major outcome of the canonical Wnt/beta-catenin-signalling pathway is the transcriptional activation of a specific set of target genes. A typical feature of the transcriptional response induced by Wnt signalling is the involvement of Tcf/Lef factors that function in the nucleus as the principal mediators of signalling. Vertebrate Tcf/Lef proteins perform two well-characterized functions: in association with beta-catenin they activate gene expression, and in the absence of Wnt ligands they bind TLE/Groucho proteins to act as transcriptional repressors. Although the general characteristics of Tcf/Lef factors are well understood, the mechanisms that control their specific roles in various cellular backgrounds are much less defined. In this report we reveal that the evolutionary conserved Dazap2 protein functions as a TCF-4 interacting partner. We demonstrate that a short region proximal to the TCF-4 HMG box mediates the interaction and that all Tcf/Lef family members associate with Dazap2. Interestingly, knockdown of Dazap2 not only reduced the activity of Wnt signalling as measured by Tcf/beta-catenin reporters but additionally altered the expression of Wnt-signalling target genes. Finally, chromatin immunoprecipitation studies indicate that Dazap2 modulates the affinity of TCF-4 for its DNA-recognition motif.

Intra-abdominal pressure correlates with abdominal wall tension during clinical evaluation tests.

BACKGROUND: The abdominal muscles play an important respiratory and stabilization role, and in coordination with other muscles regulate the intra-abdominal pressure stabilizing the spine. The evaluation of postural trunk muscle function is critical in clinical assessments of patients with musculoskeletal pain and dysfunction. This study evaluates the relationship between intra-abdominal pressure measured as anorectal pressure with objective abdominal wall tension recorded by mechanical-pneumatic-electronic sensors. METHODS: In a cross-sectional observational study, thirty-one asymptomatic participants (mean age = 26.77 ± 3.01 years) underwent testing to measure intra-abdominal pressure via anorectal manometry, along with abdominal wall tension measured by sensors attached to a trunk brace (DNS Brace). They were evaluated in five different standing postural-respiratory situations: resting breathing, Valsalva maneuver, Müller's maneuver, instructed breathing, loaded breathing when holding a dumbbell. FINDINGS: Strong correlations were demonstrated between anorectal manometry and DNS Brace measurements in all scenarios; and DNS Brace values significantly predicted intra-abdominal pressure values for all scenarios: resting breathing (r = 0.735, r2 = 0.541, p < 0.001), Valsalva maneuver (r = 0.836, r2 = 0.699, p < 0.001), Müller's maneuver (r = 0.651, r2 = 0.423, p < 0.001), instructed breathing (r = 0.708, r2 = 0.501, p < 0.001), and loaded breathing (r = 0.921, r2 = 0.848, p < 0.001). INTERPRETATION: Intra-abdominal pressure is strongly correlated with, and predicted by abdominal wall tension monitored above the inguinal ligament and in the area of superior trigonum lumbale. This study demonstrates that intra-abdominal pressure can be evaluated indirectly by monitoring the abdominal wall tension.

A Mutation Upstream of the rplN-rpsD Ribosomal Operon Downregulates Bordetella pertussis Virulence Factor Production without Compromising Bacterial Survival within Human Macrophages.

The BvgS/BvgA two-component system controls expression of ∼550 genes of Bordetella pertussis, of which, ∼245 virulence-related genes are positively regulated by the BvgS-phosphorylated transcriptional regulator protein BvgA (BvgA∼P). We found that a single G-to-T nucleotide transversion in the 5'-untranslated region (5'-UTR) of the rplN gene enhanced transcription of the ribosomal protein operon and of the rpoA gene and provoked global dysregulation of B. pertussis genome expression. This comprised overproduction of the alpha subunit (RpoA) of the DNA-dependent RNA polymerase, downregulated BvgA and BvgS protein production, and impaired production and secretion of virulence factors by the mutant. Nonetheless, the mutant survived like the parental bacteria for >2 weeks inside infected primary human macrophages and persisted within infected mouse lungs for a longer period than wild-type B. pertussis These observations suggest that downregulation of virulence factor production by bacteria internalized into host cells may enable persistence of the whooping cough agent in the airways.IMPORTANCE We show that a spontaneous mutation that upregulates transcription of an operon encoding ribosomal proteins and causes overproduction of the downstream-encoded α subunit (RpoA) of RNA polymerase causes global effects on gene expression levels and proteome composition of Bordetella pertussis Nevertheless, the resulting important downregulation of the BvgAS-controlled expression of virulence factors of the whooping cough agent did not compromise its capacity to persist for prolonged periods inside primary human macrophage cells, and it even enhanced its capacity to persist in infected mouse lungs. These observations suggest that the modulation of BvgAS-controlled expression of virulence factors may occur also during natural infections of human airways by Bordetella pertussis and may possibly account for long-term persistence of the pathogen within infected cells of the airways.

Robust method for long-term energy and pointing stabilization of high energy, high average power solid state lasers.

A robust and simple method is presented for ensuring constant energy and pointing of a high average power solid state laser on a target. In addition to providing long-term stability, this scheme also eliminates any drifts in energy or pointing resulting from the initial warm-up after a cold start. This is achieved using two separate feedback loops: one loop stabilizes the pointing of the beam external to the amplifier cavity and the other locks the cavity mode to have optimum overlap with the pump spot on the active medium. The key idea of the cavity mode stabilization is to monitor the overlap of the cavity mode and the gain medium with a camera and control it with an actively controlled, intra-cavity mirror. While this method is demonstrated on a thin-disk regenerative amplifier, it can also be applied to a wide variety of solid state laser amplifiers.

Phosphoproteomics of cAMP signaling of Bordetella adenylate cyclase toxin in mouse dendritic cells.

The adenylate cyclase toxin (CyaA) of the whooping cough agent Bordetella pertussis subverts immune functions of host myeloid cells expressing the αMß2 integrin (CD11b/CD18, CR3 or Mac-1). CyaA delivers into cytosol of cells an extremely catalytically active adenylyl cyclase enzyme, which disrupts the innate and adaptive immune functions of phagocytes through unregulated production of the key signaling molecule cAMP. We have used phosphoproteomics to analyze cAMP signaling of CyaA in murine bone marrow-derived dendritic cells. CyaA action resulted in alterations of phosphorylation state of a number of proteins that regulate actin cytoskeleton homeostasis, including Mena, Talin-1 and VASP. CyaA action repressed mTOR signaling through activation of mTORC1 inhibitors TSC2 and PRAS40 and altered phosphorylation of multiple chromatin remodelers, including the class II histone deacetylase HDAC5. CyaA toxin action further elicited inhibitory phosphorylation of SIK family kinases involved in modulation of immune response and provoked dephosphorylation of the transcriptional coactivator CRTC3, indicating that CyaA-promoted nuclear translocation of CRTC3 may account for CyaA-induced IL-10 production. These findings document the complexity of subversive physiological manipulation of myeloid phagocytes by the CyaA toxin, serving in immune evasion of the pertussis agent.

Structure-Function Relationships Underlying the Capacity of Bordetella Adenylate Cyclase Toxin to Disarm Host Phagocytes.

Bordetellae, pathogenic to mammals, produce an immunomodulatory adenylate cyclase toxin-hemolysin (CyaA, ACT or AC-Hly) that enables them to overcome the innate immune defense of the host. CyaA subverts host phagocytic cells by an orchestrated action of its functional domains, where an extremely catalytically active adenylyl cyclase enzyme is delivered into phagocyte cytosol by a pore-forming repeat-in-toxin (RTX) cytolysin moiety. By targeting sentinel cells expressing the complement receptor 3, known as the CD11b/CD18 (αMß2) integrin, CyaA compromises the bactericidal functions of host phagocytes and supports infection of host airways by Bordetellae. Here, we review the state of knowledge on structural and functional aspects of CyaA toxin action, placing particular emphasis on signaling mechanisms by which the toxin-produced 3',5'-cyclic adenosine monophosphate (cAMP) subverts the physiology of phagocytic cells.