High fidelity laser beam shaping using liquid crystal on silicon spatial light modulators as diffractive neural networks.

Spatial light modulators (SLMs) based on liquid crystal on silicon (LCoS) are powerful tools for laser beam shaping as they can be used to dynamically create almost arbitrary intensity distributions. However, laser beam shaping with LCoS-SLMs often suffers from beam shaping artifacts in part caused by unconsidered properties of the LCoS devices: astigmatism that stems from the non-normal incidence of the laser beam on the SLM and the effect commonly referred to as the '0-th diffraction order' that is caused by both the crosstalk between neighboring pixels and the direct reflection at the cover glass of the SLM. We here present a method to consider and compensate for these inherent properties of LCoS devices by treating the SLM as a diffractive neural network.

Advanced beam shaping for laser materials processing based on diffractive neural networks.

We propose a method based on neural network training algorithms for the design of diffractive neural networks - with the aim to perform advanced laser beam shaping in the NIR/VIS spectrum for laser materials processing. The method enables the efficient design of systems including multiple cascaded diffractive optical elements (DOEs) and allows the simultaneous optimization for complex (intensity and phase) target field distributions in multiple target planes. The multi-target boundary condition in the optimization method offers great potential for advanced laser beam shaping.

Freeform optics design for extended sources in paraxial approximation exploiting the expectation maximization algorithm.

Freeform optics generating specific irradiance distributions have been used in various applications for some time now. While most freeform optics design algorithms assume point sources or perfectly collimated light, the search for algorithms for non-idealized light sources with finite spatial as well as angular extent is still ongoing. In this work, such an approach is presented where the resulting irradiance distribution of a freeform optical surface is calculated as a superposition of pinhole images generated by points on the optical surface. To compute the required arrangement of the pinhole images for a prescribed irradiance pattern, the expectation maximization algorithm from statistics is applied. The result is then combined with a ray-targeting approach for finding the shape of the corresponding freeform optical surface. At its current state, the approach is applicable to Gaussian input irradiances, single-sided freeform optics and for the paraxial case. An example freeform optical surface for laser material processing is shown and discussed demonstrating the performance and the limitations of the presented approach.

Attenuate Newcastle disease virus by codon modification of the glycoproteins and phosphoprotein genes.

A codon modification strategy was used to attenuate the avian pathogenicity of an oncolytic mesogenic Newcastle disease virus (NDV) by targeting the three major virulence factors: the fusion (F) protein, hemagglutinin neuraminidase (HN) and phosphoprotein (P). Recoding the F and HN genes with rare codons greatly reduced expression of both F and HN proteins and resulted in their low incorporation into virions. The F and HN recoded virus was partially attenuated in chickens even when the F protein cleavage site was modified. Full attenuation was achieved when the 5' portion of the P gene was recoded. The recoded P, F and HN triple gene mutant exhibited delayed cell death in human cancer cells with prolonged expression of a GFP transgene. While this engineered attenuated NDV strain has lower oncolytic potency, its capacity for prolonged transgene expression may allow its use as a vaccine or gene delivery vector.

A moonlighting enzyme links Escherichia coli cell size with central metabolism.

Growth rate and nutrient availability are the primary determinants of size in single-celled organisms: rapidly growing Escherichia coli cells are more than twice as large as their slow growing counterparts. Here we report the identification of the glucosyltransferase OpgH as a nutrient-dependent regulator of E. coli cell size. During growth under nutrient-rich conditions, OpgH localizes to the nascent septal site, where it antagonizes assembly of the tubulin-like cell division protein FtsZ, delaying division and increasing cell size. Biochemical analysis is consistent with OpgH sequestering FtsZ from growing polymers. OpgH is functionally analogous to UgtP, a Bacillus subtilis glucosyltransferase that inhibits cell division in a growth rate-dependent fashion. In a striking example of convergent evolution, OpgH and UgtP share no homology, have distinct enzymatic activities, and appear to inhibit FtsZ assembly through different mechanisms. Comparative analysis of E. coli and B. subtilis reveals conserved aspects of growth rate regulation and cell size control that are likely to be broadly applicable. These include the conservation of uridine diphosphate glucose as a proxy for nutrient status and the use of moonlighting enzymes to couple growth rate-dependent phenomena to central metabolism.

Extreme C terminus of bacterial cytoskeletal protein FtsZ plays fundamental role in assembly independent of modulatory proteins.

Bacterial cell division typically requires assembly of the cytoskeletal protein FtsZ into a ring (Z-ring) at the nascent division site that serves as a foundation for assembly of the division apparatus. High resolution imaging suggests that the Z-ring consists of short, single-stranded polymers held together by lateral interactions. Several proteins implicated in stabilizing the Z-ring enhance lateral interactions between FtsZ polymers in vitro. Here we report that residues at the C terminus of Bacillus subtilis FtsZ (C-terminal variable region (CTV)) are both necessary and sufficient for stimulating lateral interactions in vitro in the absence of modulatory proteins. Swapping the 6-residue CTV from B. subtilis FtsZ with the 4-residue CTV from Escherichia coli FtsZ completely abolished lateral interactions between chimeric B. subtilis FtsZ polymers. The E. coli FtsZ chimera readily formed higher order structures normally seen only in the presence of molecular crowding agents. CTV-mediated lateral interactions are important for the integrity of the Z-ring because B. subtilis cells expressing the B. subtilis FtsZ chimera had a low frequency of FtsZ ring formation and a high degree of filamentation relative to wild-type cells. Site-directed mutagenesis of the B. subtilis CTV suggests that electrostatic forces are an important determinant of lateral interaction potential.

Actin-like protein 1 (ALP1) is a component of dynamic, high molecular weight complexes in Toxoplasma gondii.

Apicomplexan parasites, such as Toxoplasma gondii, rely on actin-based motility for cell invasion, yet conventional actin does not appear to be required for cell division in these parasites. Apicomplexans also contain a variety of actin-related proteins (Arps); however, most of these not directly orthologous to Arps in well-studied systems. We recently identified an apicomplexan-specific member of this family called Actin-Like Protein 1, (ALP1), which plays a role in the assembly of vesicular components recruited to the inner membrane complex (IMC) of daughter cells during cell division. In addition to its enrichment at daughter cell membranes, ALP1 is localized throughout the cytoplasm both diffusely distributed and concentrated in clusters that are detected by fluorescence microscopy, suggesting it forms complexes. Using quantitative optical imaging methods, including fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP), we demonstrated that ALP1 is a component of a large complex, and that it readily exchanges between diffusible and complex-bound forms. Sedimentation and density gradient analyses revealed that ALP1 is found in a freely soluble state as well as high molecular weight complexes. During cell division, ALP1 was dynamically associated with the IMC, suggesting it rapidly cycles between freely diffusible and complex forms during daughter cell assembly.