Isolation and propagation of coronaviruses in embryonated eggs.

The embryonated egg is a complex structure comprising an embryo and its supporting membranes (chorioallantoic, amniotic, yolk). The developing embryo and its membranes provide the diversity of cell types that are needed for successful replication of a wide variety of different viruses. Within the family Coronaviridae, the embryonated egg has been used as a host system primarily for two group 3 coronaviruses, infectious bronchitis virus (IBV) and turkey coronavirus (TCoV), but it also has been shown to be suitable for pheasant coronavirus.

Residues affecting the chloride regulation and substrate selectivity of the angiotensin-converting enzymes (ACE and ACE2) identified by site-directed mutagenesis.

Angiotensin-converting enzyme (ACE) and its homologue angiotensin-converting enzyme 2 (ACE2) are critical counter-regulatory enzymes of the renin-angiotensin system, and have been implicated in cardiac function, renal disease, diabetes, atherosclerosis and acute lung injury. Both ACE and ACE2 have catalytic activity that is chloride sensitive and is caused by the presence of the CL1 and CL2 chloride-binding sites in ACE and the CL1 site in ACE2. The chloride regulation of activity is also substrate dependent.

De electricitatis catholici musculari - Concerning the electrical properties of muscles, with emphasis on meat quality.

This study aims to explore the potential of evoked non-invasive surface electromyography (SEMG) analysis, in predicting meat quality traits in livestock. Evoked SEMG is a system that records, transdermally, electrical signals generated in muscle fibres upon external stimulation. These signals are reported as compound muscle action potentials (CMAP).

MRI in experimental inflammatory and mitochondrial optic neuropathies.

MRI is a powerful tool for evaluating structural and functional alterations in the optic nerve in experimental animal models of human disease. MRI-histopathological correlations have provided important insights into the pathogenesis of disease. Paramagnetic contrast agents have been used to serially visualize the foci and severity of disruption of the blood-optic nerve barrier and physiological neuronal alterations in living animals.

Array-CGH in a series of 30 patients with mental retardation, dysmorphic features, and congenital malformations detected an interstitial 1p22.2-p31.1 deletion in a patient with features overlapping the Goldenhar syndrome.

Genosensor Array 300 (Abbott) is a multiplex platform for array-based comparative genomic hybridization that detects unbalanced genomic aberrations including whole chromosome gains/losses, microdeletions, duplications and unbalanced subtelomeric rearrangements. A series of 30 patients with unexplained mental retardation, dysmorphic features, congenital abnormalities and normal high resolution karyotype and FISH subtelomeric studies were analyzed using Genosensor Array 300 array-CGH. We identified a chromosomal aberration in one patient with an interstitial 1p31.

New insights into multiple coagulation factor deficiency from the solution structure of human MCFD2.

Human MCFD2 (multiple coagulation factor deficiency 2) is a 16-kDa protein known to participate in transport of the glycosylated human coagulation factors V and VIII along the secretory pathway. Mutations in MCFD2 or in its binding partner, the membrane-bound transporter ERGIC (endoplasmic reticulum-Golgi intermediate compartment)-53, cause a mild form of inherited hemophilia known as combined deficiency of factors V and VIII (F5F8D). While ERGIC-53 is known to be a lectin-type mannose binding protein, the role of MCFD2 in the secretory pathway is comparatively unclear.

The influence of 'time since last blood meal' on the toxicity of essential oils to the poultry red mite (Dermanyssus gallinae).

The poultry red mite, Dermanyssus gallinae (De Geer) is a serious ectoparasitic pest of layer hens that can survive for long periods in the poultry house sub-structure without taking a blood meal from its host. The research undertaken in this study found that 'time since last blood meal' had a notable effect on how toxic a selection of plant essential oils were to D. gallinae under laboratory conditions.

Optic nerve degeneration in experimental autoimmune encephalomyelitis.

The mechanisms of axonal and neuronal degeneration causing disability in optic neuritis and multiple sclerosis are poorly understood. Here we describe the role of mitochondria, oxidative stress and the effects of modulating antioxidant gene expression in the optic nerves of mice induced with experimental autoimmune encephalomyelitis, with a focus on long-term neuroprotection. Oxidative injury to the mitochondrion began prior to inflammatory cell infiltration and continued.

Effect of reducing dietary mineral content on growth performance, water intake, excreta dry matter content and blood parameters of broilers.

1. The objective of this study was to investigate whether it is possible to reduce phosphorus (P) excretion by feeding a lower level of dietary P than that currently recommended, without compromising welfare, and whether this is sensitive to the level of dietary calcium (Ca). 2.

Lack of prolonged activity of lavender essential oils as acaricides against the poultry red mite (Dermanyssusgallinae) under laboratory conditions.

Managing the poultry red mite, Dermanyssusgallinae (De Geer) by conventional means (i.e., synthetic acaricides) has become increasingly problematic.

Functional angiotensin-converting enzyme 2 is expressed in human cardiac myofibroblasts.

The renin-angiotensin system (RAS), in particular angiotensin II, plays an important role in cardiac remodelling. Angiotensin-converting enzyme (ACE) and angiotensin-converting enzyme 2 (ACE2) are key players in the RAS and act antagonistically to regulate the levels of angiotensin II. In this study, we reveal the functional expression of ACE2 in human cardiac myofibroblasts, cells that are essential to the maintenance of normal cardiac architecture and also play a key role in myocardial remodelling.

Growth modelling indicates hurricanes and severe storms are linked to low coral recruitment in the Caribbean.

This study set out to test the hypothesis that hurricanes and tropical storms limit the recruitment and subsequent survival of massive non-branching corals on the barrier reef off the coast of Belize in the Gulf of Honduras. Overall, the surface areas of 523 individual coral specimens were measured, and recruitment dates were then modelled. There was no significant difference in coral cover or coral biodiversity between any of the sites studied (p > 0.

Long-term suppression of neurodegeneration in chronic experimental optic neuritis: antioxidant gene therapy.

Purpose: To test in mice with experimental autoimmune encephalomyelitis (EAE) a strategy designed to treat patients at risk for axonal degeneration and persistent visual loss from optic neuritis and multiple sclerosis.

Methods: The authors cloned the human extracellular superoxide dismutase (ECSOD) or catalase (CAT) gene into recombinant adenoassociated virus (AAV). Transgene expression was evaluated by immunochemistry of infected RGC-5 cells and after intravitreal injection of AAV-ECSOD or AAV-CAT, or both, into the right eyes of DBA/1J mice.

Natural methods of protein stabilization: thermostable biocatalysts.

Enzymes that are naturally found in thermophilic and hyperthermophilic organisms are being used as robust biocatalysts in the fine chemical and pharmaceutical industries. They have important use in these industries due to their increased stability which is often required during commercial reaction conditions. The approach used in these studies is to learn how nature has managed to stabilize these proteins using a detailed knowledge of their biochemical properties and three-dimensional structures.

Cone-specific expression using a human red opsin promoter in recombinant AAV.

Purpose: To determine the feasibility of targeting gene expression specifically to cone photoreceptors using recombinant adeno-associated virus (rAAV) as the vector.

Methods: An rAAV vector was constructed that contains a 2.1kb upstream sequence of the human red opsin gene to direct green fluorescent protein (GFP) expression.

Convergent preparation and photophysical characterization of dimaleimide dansyl fluorogens: elucidation of the maleimide fluorescence quenching mechanism.

Dimaleimide fluorogens are being developed for application to fluorescent protein labeling. In this method, fluorophores bearing two maleimide quenching groups do not fluoresce until both maleimide groups have undergone thiol addition reactions with the Cys residues of the target protein sequence [J. Am.

Genomic characterization of equine coronavirus.

The complete genome sequence of the first equine coronavirus (ECoV) isolate, NC99 strain was accomplished by directly sequencing 11 overlapping fragments which were RT-PCR amplified from viral RNA. The ECoV genome is 30,992 nucleotides in length, excluding the polyA tail. Analysis of the sequence identified 11 open reading frames which encode two replicase polyproteins, five structural proteins (hemagglutinin esterase, spike, envelope, membrane, and nucleocapsid) and four accessory proteins (NS2, p4.

Experimental evaluation of Musca domestica (Diptera: Muscidae) as a vector of Newcastle disease virus.

House flies, Musca domestica L. (Diptera: Muscidae), were examined for their ability to harbor and transmit Newcastle disease virus (family Paramyxoviridae, genus Avulavirus, NDV) by using a mesogenic NDV strain. Laboratory-reared flies were experimentally exposed to NDV (Roakin strain) by allowing flies to imbibe an inoculum consisting of chicken embryo-propagated virus.

A titer plate-based polymer microfluidic platform for high throughput nucleic acid purification.

A 96-well solid-phase reversible immobilization (SPRI) reactor plate was designed to demonstrate functional titer plate-based microfluidic platforms. Nickel, large area mold inserts were fabricated using an SU-8 based, UV-LIGA technique on 150 mm diameter silicon substrates. Prior to UV exposure, the prebaked SU-8 resist was flycut to reduce the total thickness variation to less than 5 mum.

The crystal structure of the ivy Delta4-16:0-ACP desaturase reveals structural details of the oxidized active site and potential determinants of regioselectivity.

The multifunctional acyl-acyl carrier protein (ACP) desaturase from Hedera helix (English ivy) catalyzes the Delta(4) desaturation of 16:0-ACP and the Delta(9) desaturation of 18:0-ACP and further desaturates Delta(9)-16:1 or Delta(9)-18:1 to the corresponding Delta(4,9) dienes. The crystal structure of the enzyme has been solved to 1.95 A resolution, and both the iron-iron distance of approximately 3.

Experimental reproduction of transmissible viral proventriculitis by infection of chickens with a novel adenovirus-like virus (isolate R11/3).

Transmissible viral proventriculitis (TVP) was experimentally reproduced in 2-wk-old specific-pathogen-free chickens and commercial broiler chickens by eyedrop inoculation of adenovirus-like virus (AdLV), isolate R1 1/3. No clinical signs and no weight gain depression were observed in chickens inoculated with AdLV (R11/3); however, gross and microscopic lesions characteristic of TVP were present in proventriculi of inoculated chickens. Proventriculi of AdLV (R11/3)-inoculated chickens were markedly enlarged, compared with sham-inoculated controls, by day 7 postinoculation (PI).

Use of mitochondrial antioxidant defenses for rescue of cells with a Leber hereditary optic neuropathy-causing mutation.

Objective: To explore a treatment paradigm for Leber hereditary optic neuropathy (LHON), we augmented mitochondrial antioxidant defenses to rescue cells with the G11778A mutation in mitochondrial DNA.

Methods: Cells homoplasmic for the G11778A mutation in mitochondrial DNA were infected with an adeno-associated viral vector containing the human mitochondrial superoxide dismutase (SOD2) gene. Control cells were infected with an adeno-associated viral (AAV) vector expressing the green fluorescent protein (GFP).