PCR detection and analysis of Pasteurella multocida from the tonsils of slaughtered pigs in Vietnam

A total of 36 tonsil swab samples were collected from healthy swine prior to slaughter at the abattoirs in Can tho and Tien giang provinces of Southern Vietnam, The presence of Pasteurella multocida in these samples was detected by the combination of direct cultivation and isolation, mouse inoculation and the polymerase chain reaction (PM-PCR). P. multocida was detected in 16 samples by PCR, with 17 strains ultimately isolated. All samples were negative for serogroup B by HSB-PCR and conventional serotyping, with isolates identified as A:3, D:1 or D:3. In addition, all samples were determined to be negative for the P. multocida toxin (PMT). Characterisation of isolated P, multocida by REP-PCR and biotyping revealed nine distinct REP profiles and seven biotypes among the 17 isolates. Some correlation was seen with P. multocida isolated from a previous Australian outbreak of acute swine pasteurellosis, and those isolated from fowl cholera outbreaks in Vietnamese poultry. (C) 2000 Elsevier Science B.V. All rights reserved.

XSophe, a computer simulation software suite for the analysis of electron paramagnetic resonance spectra

The XSophe computer simulation software suite consisting of a daemon, the XSophe interface and the computational program Sophe is a state of the art package for the simulation of electron paramagnetic resonance spectra. The Sophe program performs the computer simulation and includes a number of new technologies including; the SOPHE partition and interpolation schemes, a field segmentation algorithm, homotopy, parallelisation and spectral optimisation. The SOPHE partition and interpolation scheme along with a field segmentation algorithm greatly increases the speed of simulations for most systems. Multidimensional homotopy provides an efficient method for accurately tracing energy levels and hence tracing transitions in the presence of energy level anticrossings and looping transitions and allowing computer simulations in frequency space. Recent enhancements to Sophe include the generalised treatment of distributions of orientational parameters, termed the mosaic misorientation linewidth model and a faster more efficient algorithm for the calculation of resonant field positions and transition probabilities. For complex systems the parallelisation enables the simulation of these systems on a parallel computer and the optimisation algorithms in the suite provide the experimentalist with the possibility of finding the spin Hamiltonian parameters in a systematic manner rather than a trial-and-error process. The XSophe software suite has been used to simulate multifrequency EPR spectra (200 MHz to 6 00 GHz) from isolated spin systems (S > ~½) and coupled centres (Si, Sj _> I/2). Griffin, M.; Muys, A.; Noble, C.; Wang, D.; Eldershaw, C.; Gates, K.E.; Burrage, K.; Hanson, G.R."XSophe, a Computer Simulation Software Suite for the Analysis of Electron Paramagnetic Resonance Spectra", 1999, Mol. Phys. Rep., 26, 60-84.

Controlling contractual discretion

In 1993 Leggatt L.J. said in Abu Dhabi National Tanker Co. v. Product Star Shipping Ltd. (The Product Star) [1993] 1 Lloyds Rep. 397, 404: Where A and B contract with each other to confer a discretion on A, that does not render B subject to A’s uninhibited whim. In my judgment, the authorities show that not only must the discretion be exercised honestly and in good faith, but, having regard to the provisions of the contract by which it is conferred, it must not be exercised arbitrarily, capriciously or unreasonably.

Improving distributed representation of word sense via WordNet Gloss composition and context clustering

In recent years, there has been an increas-ing interest in learning a distributed rep-resentation of word sense. Traditional context clustering based models usually require careful tuning of model parame-ters, and typically perform worse on infre-quent word senses. This paper presents a novel approach which addresses these lim-itations by first initializing the word sense embeddings through learning sentence-level embeddings from WordNet glosses using a convolutional neural networks. The initialized word sense embeddings are used by a context clustering based model to generate the distributed representations of word senses. Our learned represen-tations outperform the publicly available embeddings on 2 out of 4 metrics in the word similarity task, and 6 out of 13 sub tasks in the analogical reasoning task.

Some Fixed Point Theorems for Kannan Mappings

2000 Mathematics Subject Classification: Primary: 47H10; Secondary: 54H25.

Surface microbial consortia from Livarot, a French smear-ripened cheese

The surface microflora (902 isolates) of Livarot cheeses from three dairies was investigated during ripening. Yeasts were mainly identified by Fourier transform infrared spectroscopy. Geotrichum candidum was the dominating yeast among 10 species. Bacteria were identified using Biotype 100 strips, dereplicated by repetitive extragenic palindromic PCR (rep-PCR); 156 representative strains were identified by either BOX-PCR or (GTG) 55-PCR, and when appropriate by 16S rDNA sequencing and SDS-PAGE analysis. Gram-positive bacteria accounted for 65% of the isolates and were mainly assigned to the genera Arthrobacter, Brevibacterium, Corynebacterium, and Staphylococcus. New taxa related to the genera Agrococcus and Leucobacter were found. Yeast and Gram-positive bacteria strains deliberately added as smearing agents were sometimes undetected during ripening. Thirty-two percent of the isolates were Gram-negative bacteria, which showed a high level of diversity and mainly included members of the genera Alcaligenes, Hafnia, Proteus, Pseudomonas, and Psychrobacter. Whatever the milk used (pasteurized or unpasteurized), similar levels of biodiversity were observed in the three dairies, all of which had efficient cleaning procedures and good manufacturing practices. It appears that some of the Gramnegative bacteria identified should now be regarded as potentially useful in some cheese technologies. The assessment of their positive versus negative role should be objectively examined.

Biodiversity of the surface microbial consortia from Limburger, Reblochon, Livarot, Tilsit, and Gubbeen cheeses

Comprehensive collaborative studies from our laboratories reveal the extensive biodiversity of the microflora of the surfaces of smear-ripened cheeses. Two thousand five hundred ninety-seven strains of bacteria and 2,446 strains of yeasts from the surface of the smear-ripened cheeses Limburger, Reblochon, Livarot, Tilsit, and Gubbeen, isolated at three or four times during ripening, were identified; 55 species of bacteria and 30 species of yeast were found. The microfloras of the five cheeses showed many similarities but also many differences and interbatch variation. Very few of the commercial smear microorganisms, deliberately inoculated onto the cheese surface, were reisolated and then mainly from the initial stages of ripening, implying that smear cheese production units must have an adventitious "house" flora. Limburger cheese had the simplest microflora, containing two yeasts, Debaryomyces hansenii and Geotrichum candidum, and two bacteria, Arthrobacter arilaitensis and Brevibacterium aurantiacum. The microflora of Livarot was the most complicated, comprising 10 yeasts and 38 bacteria, including many gram-negative organisms. Reblochon also had a very diverse microflora containing 8 yeasts and 13 bacteria (excluding gram-negative organisms which were not identified), while Gubbeen had 7 yeasts and 18 bacteria and Tilsit had 5 yeasts and 9 bacteria. D. hansenii was by far the dominant yeast, followed in order by G. candidum, Candida catenulata, and Kluyveromyces lactis. B. aurantiacum was the dominant bacterium and was found in every batch of the 5 cheeses. The next most common bacteria, in order, were Staphylococcus saprophyticus, A. arilaitensis, Corynebacterium casei, Corynebacterium variabile, and Microbacterium gubbeenense. S. saprophyticus was mainly found in Gubbeen, and A. arilaitensis was found in all cheeses but not in every batch. C. casei was found in most batches of Reblochon, Livarot, Tilsit, and Gubbeen. C. variabile was found in all batches of Gubbeen and Reblochon but in only one batch of Tilsit and in no batch of Limburger or Livarot. Other bacteria were isolated in low numbers from each of the cheeses, suggesting that each of the 5 cheeses has a unique microflora. In Gubbeen cheese, several different strains of the dominant bacteria were present, as determined by pulsed-field gel electrophoresis, and many of the less common bacteria were present as single clones. The culture-independent method, denaturing gradient gel electrophoresis, resulted in identification of several bacteria which were not found by the culture-dependent (isolation and rep-PCR identification) method. It was thus a useful complementary technique to identify other bacteria in the cheeses. The gross composition, the rate of increase in pH, and the indices of proteolysis were different in most of the cheeses.

Control of the in-cavity dynamics in mode-locked fibre lasers

At the level of fundamental research, fibre lasers provide convenient and reproducible experimental settings for the study of a variety of nonlinear dynamical processes, while at the applied research level, pulses with different and optimised features – e.g., in terms of pulse duration, temporal and/or spectral intensity profile, energy, repetition rate and emission bandwidth – are sought with the general constraint of developing efficient cavity architectures. In this talk, we review our recent progress on the realisation of different regimes of pulse generation in passively mode-locked fibre lasers through control of the in-cavity propagation dynamics. We report on the possibility to achieve both parabolic self-similar and triangular pulse shaping in a mode-locked fibre laser via adjustment of the net normal dispersion and integrated gain of the cavity [1]. We also show that careful control of the gain/loss parameters of a net-normal dispersion laser cavity provides the means of achieving switching among Gaussian pulse, dissipative soliton and similariton pulse solutions in the cavity [2,3]. Furthermore, we report on our recent theoretical and experimental studies of pulse shaping by inclusion of an amplitude and phase spectral filter into the cavity of a laser. We numerically demonstrate that a mode-locked fibre laser can operate in dif- ferent pulse-generation regimes, including parabolic, flattop and triangular waveform generations, depending on the amplitude profile of the in-cavity spectral filter [4]. An application of technique using a flat-top spectral filter is demonstrated to achieve the direct generation of sinc-shaped optical Nyquist pulses of high quality and of a widely tuneable bandwidth from the laser [5]. We also report on a recently-developed versa- tile erbium-doped fibre laser, in which conventional soliton, dispersion-managed soli- ton (stretched-pulse) and dissipative soliton mode-locking regimes can be selectively and reliably targeted by programming different group-velocity dispersion profiles and bandwidths on an in-cavity programmable filter [6]. References: 1. S. Boscolo and S. K. Turitsyn, Phys. Rev. A 85, 043811 (2012). 2. J. Peng et al., Phys. Rev. A 86, 033808 (2012). 3. J. Peng, Opt. Express 24, 3046-3054 (2016). 4. S. Boscolo, C. Finot, H. Karakuzu, and P. Petropoulos, Opt. Lett. 39, 438-441 (2014). 5. S. Boscolo, C. Finot, and S. K. Turitsyn, IEEE Photon. J. 7, 7802008 (2015). 6. J. Peng and S. Boscolo, Sci. Rep. 6, 25995 (2016).

In-cavity pulse shaping and dissipative parametric instability mode-locking in fibre lasers

At the level of fundamental research, fibre lasers provide convenient and reproducible experimental settings for the study of a variety of nonlinear dynamical processes, while at the applied research level, pulses with different and optimised features – e.g., in terms of pulse duration, temporal and/or spectral shape, energy, repetition rate and emission bandwidth – are sought with the general constraint of developing efficient cavity architectures. In this work, we review our recent progress on the realisation of pulse shaping in passively- mode-locked fibre lasers by inclusion of an amplitude and phase spectral filter into the laser cavity. We present a fibre laser design in which pulse shaping occurs through filtering of a spectrally nonlinearly broadened pulse in the cavity. This strategy of pulse shaping is illustrated through the numerical demonstration of the laser operation in different pulse-generation regimes, including parabolic, flattop and triangular waveform generations, depending on the amplitude profile of the in-cavity spectral filter [1]. As an application of this general approach, we show that the use of an in-cavity flat-top spectral filter makes it possible to directly generate sinc-shaped Nyquist pulses of high quality and of a widely tunable bandwidth from the laser [2]. We also report on a recently-developed versatile erbium-doped fibre laser, in which conventional soliton, dispersion-managed soliton (stretched-pulse) and dissipative soliton mode-locking regimes can be selectively and reliably targeted by programming different group-velocity dispersion profiles and bandwidths on an in-cavity programmable filter [3]. Further, we report on our recent results on the passive mode locking of a Raman fibre laser by a recently predicted new type of parametric instability – the dissipative Faraday instability [4], where spatially periodic zig-zag modulation of spectrally dependent losses can lead to pattern formation in the temporal domain. High-order harmonic mode locking is achieved in a very simple experimental configuration, with the laser cavity including an optical fibre and two chirped fibre Bragg gratings, and no additional mode-locking elements. The results not only open up new possibilities for the design of mode-locked lasers, but extend beyond fibre optics to other fields of physics and engineering. References [1] S. Boscolo, C. Finot, H. Karakuzu, P. Petropoulos, “Pulse shaping in mode-locked fiber laser by in-cavity spectral filter,” Opt. Lett., vol. 39, pp. 438–441, 2014. [2] S. Boscolo, C. Finot, S. K. Turitsyn, “Bandwidth programmable optical Nyquist pulse generation in passively mode-locked fiber laser,” IEEE Photon. J., vol. 7, 7802008(8), 2015. [3] J. Peng, S. Boscolo, “Filter-based dispersion-managed versatile ultrafast fibre laser,” Sci. Rep., 2016, In press. [4] A. M. Perego, N. Tarasov, D. V. Churkin, S. K. Turitsyn, K. Staliunas, “Pattern generation by dissipative parametric instability,” Phys. Rev. Lett., vol. 116, 028701, 2016.

Carnival's Fantasy Class Cruise Ship: Ticket to Success

Carnival Cruise Line's Fantasy class of cruise ships is the largest group of virtually identical passenger vessels in the history of ocean travel. These ships rep- resent the culmination of Carnival's product development and are a prime reason for the line's current success. The author details the evolution of their design, with emphasis on hotel aspects, through previous ships in the fleet.

A Therapeutic Antibody for Cancer, Derived from Single Human B Cells.

Some patients with cancer never develop metastasis, and their host response might provide cues for innovative treatment strategies. We previously reported an association between autoantibodies against complement factor H (CFH) and early-stage lung cancer. CFH prevents complement-mediated cytotoxicity (CDC) by inhibiting formation of cell-lytic membrane attack complexes on self-surfaces. In an effort to translate these findings into a biologic therapy for cancer, we isolated and expressed DNA sequences encoding high-affinity human CFH antibodies directly from single, sorted B cells obtained from patients with the antibody. The co-crystal structure of a CFH antibody-target complex shows a conformational change in the target relative to the native structure. This recombinant CFH antibody causes complement activation and release of anaphylatoxins, promotes CDC of tumor cell lines, and inhibits tumor growth in vivo. The isolation of anti-tumor antibodies derived from single human B cells represents an alternative paradigm in antibody drug discovery.

Small molecule dual-inhibitors of TRPV4 and TRPA1 for attenuation of inflammation and pain.

TRPV4 ion channels represent osmo-mechano-TRP channels with pleiotropic function and wide-spread expression. One of the critical functions of TRPV4 in this spectrum is its involvement in pain and inflammation. However, few small-molecule inhibitors of TRPV4 are available. Here we developed TRPV4-inhibitory molecules based on modifications of a known TRPV4-selective tool-compound, GSK205. We not only increased TRPV4-inhibitory potency, but surprisingly also generated two compounds that potently co-inhibit TRPA1, known to function as chemical sensor of noxious and irritant signaling. We demonstrate TRPV4 inhibition by these compounds in primary cells with known TRPV4 expression - articular chondrocytes and astrocytes. Importantly, our novel compounds attenuate pain behavior in a trigeminal irritant pain model that is known to rely on TRPV4 and TRPA1. Furthermore, our novel dual-channel blocker inhibited inflammation and pain-associated behavior in a model of acute pancreatitis - known to also rely on TRPV4 and TRPA1. Our results illustrate proof of a novel concept inherent in our prototype compounds of a drug that targets two functionally-related TRP channels, and thus can be used to combat isoforms of pain and inflammation in-vivo that involve more than one TRP channel. This approach could provide a novel paradigm for treating other relevant health conditions.

Scaffold-free, Human Mesenchymal Stem Cell-Based Tissue Engineered Blood Vessels.

Tissue-engineered blood vessels (TEBV) can serve as vascular grafts and may also play an important role in the development of organs-on-a-chip. Most TEBV construction involves scaffolding with biomaterials such as collagen gel or electrospun fibrous mesh. Hypothesizing that a scaffold-free TEBV may be advantageous, we constructed a tubular structure (1 mm i.d.) from aligned human mesenchymal cell sheets (hMSC) as the wall and human endothelial progenitor cell (hEPC) coating as the lumen. The burst pressure of the scaffold-free TEBV was above 200 mmHg after three weeks of sequential culture in a rotating wall bioreactor and perfusion at 6.8 dynes/cm(2). The interwoven organization of the cell layers and extensive extracellular matrix (ECM) formation of the hMSC-based TEBV resembled that of native blood vessels. The TEBV exhibited flow-mediated vasodilation, vasoconstriction after exposure to 1 μM phenylephrine and released nitric oxide in a manner similar to that of porcine femoral vein. HL-60 cells attached to the TEBV lumen after TNF-α activation to suggest a functional endothelium. This study demonstrates the potential of a hEPC endothelialized hMSC-based TEBV for drug screening.

Direct In Vivo Manipulation and Imaging of Calcium Transients in Neutrophils Identify a Critical Role for Leading-Edge Calcium Flux.

Calcium signaling has long been associated with key events of immunity, including chemotaxis, phagocytosis, and activation. However, imaging and manipulation of calcium flux in motile immune cells in live animals remain challenging. Using light-sheet microscopy for in vivo calcium imaging in zebrafish, we observe characteristic patterns of calcium flux triggered by distinct events, including phagocytosis of pathogenic bacteria and migration of neutrophils toward inflammatory stimuli. In contrast to findings from ex vivo studies, we observe enriched calcium influx at the leading edge of migrating neutrophils. To directly manipulate calcium dynamics in vivo, we have developed transgenic lines with cell-specific expression of the mammalian TRPV1 channel, enabling ligand-gated, reversible, and spatiotemporal control of calcium influx. We find that controlled calcium influx can function to help define the neutrophil's leading edge. Cell-specific TRPV1 expression may have broad utility for precise control of calcium dynamics in other immune cell types and organisms.

Live volumetric (4D) visualization and guidance of in vivo human ophthalmic surgery with intraoperative optical coherence tomography.

Minimally-invasive microsurgery has resulted in improved outcomes for patients. However, operating through a microscope limits depth perception and fixes the visual perspective, which result in a steep learning curve to achieve microsurgical proficiency. We introduce a surgical imaging system employing four-dimensional (live volumetric imaging through time) microscope-integrated optical coherence tomography (4D MIOCT) capable of imaging at up to 10 volumes per second to visualize human microsurgery. A custom stereoscopic heads-up display provides real-time interactive volumetric feedback to the surgeon. We report that 4D MIOCT enhanced suturing accuracy and control of instrument positioning in mock surgical trials involving 17 ophthalmic surgeons. Additionally, 4D MIOCT imaging was performed in 48 human eye surgeries and was demonstrated to successfully visualize the pathology of interest in concordance with preoperative diagnosis in 93% of retinal surgeries and the surgical site of interest in 100% of anterior segment surgeries. In vivo 4D MIOCT imaging revealed sub-surface pathologic structures and instrument-induced lesions that were invisible through the operating microscope during standard surgical maneuvers. In select cases, 4D MIOCT guidance was necessary to resolve such lesions and prevent post-operative complications. Our novel surgical visualization platform achieves surgeon-interactive 4D visualization of live surgery which could expand the surgeon's capabilities.