Morphological and Molecular Characteristics of <i>Sarcocystis aucheniae</i> Isolated from Meat of Guanaco (<i>Lama guanicoe</i>)

Background: Sarcocystosis in South American camelids (SAC) is an important parasitic disease which results in economical loss due to carcass condemnation. Meat products from camelids are significant source of animal protein in several American countries. Sarcocystis spp. producing macroscopical cysts in these animals have been nominated as S. aucheniae, S. tilopodi, and S. guanicoecanis. The aim of the present study was to characterize morphologically and molecularly Sarcocystis spp. producing macroscopical cysts in guanacos (Lama guanicoe). Methods: Samples of macroscopical cysts were dissected from guanaco muscles slaughtered in Santa Cruz province, Argentina. The samples were processed by transmission electron microscopy, polymerase chain reaction (PCR), cloning and sequencing studies. Results: Cyst wall villar protrusions were irregular with a “tree” or “cauliflower-like” shape which measured 3-4.5 µm height by 2.5-3.5 µm width. A consensus 18S rRNA full gene sequence of 1875 bp was obtained and showed the highest identity (≥ 99 %) with sequences of S. aucheniae reported in GenBank. The ultra structure of the cyst wall from macroscopical cysts and 18S rRNA gene obtained from guanacos were identical and consistent with the descriptions of S. aucheniae. Conclusion: This study confirms that guanacos are infected by the same Sarcocystis spp. affecting other SAC, named S. aucheniae, and reinforce the suggestion that nomination of new species should contain morphological and molecular data to reach validity. In future, this data could be useful to obtain proper identifications in post-mortem inspection and helpful to improve meat safety.Facultad de Ciencias Veterinaria

Morphological and Molecular Characteristics of <i>Sarcocystis aucheniae</i> Isolated from Meat of Guanaco (<i>Lama guanicoe</i>)

Background: Sarcocystosis in South American camelids (SAC) is an important parasitic disease which results in economical loss due to carcass condemnation. Meat products from camelids are significant source of animal protein in several American countries. Sarcocystis spp. producing macroscopical cysts in these animals have been nominated as S. aucheniae, S. tilopodi, and S. guanicoecanis. The aim of the present study was to characterize morphologically and molecularly Sarcocystis spp. producing macroscopical cysts in guanacos (Lama guanicoe). Methods: Samples of macroscopical cysts were dissected from guanaco muscles slaughtered in Santa Cruz province, Argentina. The samples were processed by transmission electron microscopy, polymerase chain reaction (PCR), cloning and sequencing studies. Results: Cyst wall villar protrusions were irregular with a “tree” or “cauliflower-like” shape which measured 3-4.5 µm height by 2.5-3.5 µm width. A consensus 18S rRNA full gene sequence of 1875 bp was obtained and showed the highest identity (≥ 99 %) with sequences of S. aucheniae reported in GenBank. The ultra structure of the cyst wall from macroscopical cysts and 18S rRNA gene obtained from guanacos were identical and consistent with the descriptions of S. aucheniae. Conclusion: This study confirms that guanacos are infected by the same Sarcocystis spp. affecting other SAC, named S. aucheniae, and reinforce the suggestion that nomination of new species should contain morphological and molecular data to reach validity. In future, this data could be useful to obtain proper identifications in post-mortem inspection and helpful to improve meat safety.Facultad de Ciencias Veterinaria

Optical mesh lattices with PT-symmetry

We investigate a new class of optical mesh periodic structures that are discretized in both the transverse and longitudinal directions. These networks are composed of waveguide arrays that are discretely coupled while phase elements are also inserted to discretely control their effective potentials and can be realized both in the temporal and the spatial domain. Their band structure and impulse response is studied in both the passive and parity-time (PT) symmetric regime. The possibility of band merging and the emergence of exceptional points along with the associated optical dynamics are considered in detail both above and below the PT-symmetry breaking point. Finally unidirectional invisibility in PT-synthetic mesh lattices is also examined along with possible superluminal light transport dynamics.Comment: 14 pages, 17 figures, published in Physical Review

Recursive Definitions of Monadic Functions

Using standard domain-theoretic fixed-points, we present an approach for defining recursive functions that are formulated in monadic style. The method works both in the simple option monad and the state-exception monad of Isabelle/HOL's imperative programming extension, which results in a convenient definition principle for imperative programs, which were previously hard to define. For such monadic functions, the recursion equation can always be derived without preconditions, even if the function is partial. The construction is easy to automate, and convenient induction principles can be derived automatically.Comment: In Proceedings PAR 2010, arXiv:1012.455

An integrated biochemical system for nitrate assimilation and nitric oxide detoxification in Bradyrhizobium japonicum

Rhizobia are recognized to establish N(2)-fixing symbiotic interactions with legume plants. Bradyrhizobium japonicum, the symbiont of soybeans, can denitrify and grow under free-living conditions with nitrate (NO(3)(−)) or nitrite (NO(2)(−)) as sole nitrogen source. Unlike related bacteria that assimilate NO(3)(−), genes encoding the assimilatory NO(3)(−) reductase (nasC) and NO(2)(−) reductase (nirA) in B. japonicum are located at distinct chromosomal loci. The nasC gene is located with genes encoding an ABC-type NO(3)(−) transporter, a major facilitator family NO(3)(−)/NO(2)(−) transporter (NarK), flavoprotein (Flp) and single-domain haemoglobin (termed Bjgb). However, nirA clusters with genes for a NO(3)(−)/NO(2)(−)-responsive regulator (NasS-NasT). In the present study, we demonstrate NasC and NirA are both key for NO(3)(−) assimilation and that growth with NO(3)(−), but not NO(2)(−) requires flp, implying Flp may function as electron donor to NasC. In addition, bjgb and flp encode a nitric oxide (NO) detoxification system that functions to mitigate cytotoxic NO formed as a by-product of NO(3)(−) assimilation. Additional experiments reveal NasT is required for NO(3)(−)-responsive expression of the narK-bjgb-flp-nasC transcriptional unit and the nirA gene and that NasS is also involved in the regulatory control of this novel bipartite assimilatory NO(3)(−)/NO(2)(−) reductase pathway

Optoacoustic Imaging in Inflammation.

Optoacoustic or photoacoustic imaging (OAI/PAI) is a technology which enables non-invasive visualization of laser-illuminated tissue by the detection of acoustic signals. The combination of "light in" and "sound out" offers unprecedented scalability with a high penetration depth and resolution. The wide range of biomedical applications makes this technology a versatile tool for preclinical and clinical research. Particularly when imaging inflammation, the technology offers advantages over current clinical methods to diagnose, stage, and monitor physiological and pathophysiological processes. This review discusses the clinical perspective of using OAI in the context of imaging inflammation as well as in current and emerging translational applications

Composing and Factoring Generalized Green's Operators and Ordinary Boundary Problems

We consider solution operators of linear ordinary boundary problems with "too many" boundary conditions, which are not always solvable. These generalized Green's operators are a certain kind of generalized inverses of differential operators. We answer the question when the product of two generalized Green's operators is again a generalized Green's operator for the product of the corresponding differential operators and which boundary problem it solves. Moreover, we show that---provided a factorization of the underlying differential operator---a generalized boundary problem can be factored into lower order problems corresponding to a factorization of the respective Green's operators. We illustrate our results by examples using the Maple package IntDiffOp, where the presented algorithms are implemented.Comment: 19 page

An improved method for high-throughput quantification of autophagy in mammalian cells

Autophagy is a cellular homeostatic pathway with functions ranging from cytoplasmic protein turnover to immune defense. Therapeutic modulation of autophagy has been demonstrated to positively impact the outcome of autophagy-dysregulated diseases such as cancer or microbial infections. However, currently available agents lack specificity, and new candidates for drug development or potential cellular targets need to be identified. Here, we present an improved method to robustly detect changes in autophagy in a high-throughput manner on a single cell level, allowing effective screening. This method quantifies eGFP-LC3B positive vesicles to accurately monitor autophagy. We have significantly streamlined the protocol and optimized it for rapid quantification of large numbers of cells in little time, while retaining accuracy and sensitivity. Z scores up to 0.91 without a loss of sensitivity demonstrate the robustness and aptness of this approach. Three exemplary applications outline the value of our protocols and cell lines: (I) Examining autophagy modulating compounds on four different cell types. (II) Monitoring of autophagy upon infection with e.g. measles or influenza A virus. (III) CRISPR/Cas9 screening for autophagy modulating factors in T cells. In summary, we offer ready-to-use protocols to generate sensitive autophagy reporter cells and quantify autophagy in high-throughput assays