Assembly and movement of a plant virus carrying a green fluorescent protein overcoat.

Potato virus X (PVX) is a filamentous plant virus infecting many members of the family Solanaceae. A modified form of PVX, PVX.GFP-CP which expressed a chimeric gene encoding a fusion between the 27-kDa Aequorea victoria green fluorescent protein and the amino terminus of the 25-kDa PVX coat protein, assembled into virions and moved both locally and systemically. The PVX.GFP-CP virions were over twice the diameter of wild-type PVX virions. Assembly of PVX.GFP-CP virions required the presence of free coat protein subunits in addition to the fusion protein subunits. PVX.GFP-CP virions accumulated as paracrystalline arrays in infected cells similar to those seen in cells infected with wild-type PVX The formation of virions carrying large superficial fusions illustrates a novel approach for production of high levels of foreign proteins in plants. Aggregates of PVX.GFP-CP particles were fluorescent, emitting green light when excited with ultraviolet light and could be imaged using confocal laser scanning microscopy. The detection of virus particles in infected tissue demonstrates the potential of fusions between the green fluorescent protein and virus coat protein for the non-invasive study of virus multiplication and spread.

Crystal structures of the trimeric human immunodeficiency virus type 1 matrix protein: implications for membrane association and assembly.

The human immunodeficiency virus type 1 (HIV-1) matrix protein forms a structural shell associated with the inner viral membrane and performs other essential functions throughout the viral life cycle. The crystal structure of the HIV-1 matrix protein, determined at 2.3 angstrom resolution, reveals that individual matrix molecules are composed of five major helices capped by a three-stranded mixed beta-sheet. Unexpectedly, the protein assembles into a trimer in three different crystal lattices, burying 1880 angstrom2 of accessible surface area at the trimer interfaces. Trimerization appears to create a large, bipartite membrane binding surface in which exposed basic residues could cooperate with the N-terminal myristoyl groups to anchor the protein on the acidic inner membrane of the virus.

A mean field model of ligand-protein interactions: implications for the structural assessment of human immunodeficiency virus type 1 protease complexes and receptor-specific binding.

We propose a general mean field model of ligand-protein interactions to determine the thermodynamic equilibrium of a system at finite temperature. The method is employed in structural assessments of two human immuno-deficiency virus type 1 protease complexes where the gross effects of protein flexibility are incorporated by utilizing a data base of crystal structures. Analysis of the energy spectra for these complexes has revealed that structural and thermo-dynamic aspects of molecular recognition can be rationalized on the basis of the extent of frustration in the binding energy landscape. In particular, the relationship between receptor-specific binding of these ligands to human immunodeficiency virus type 1 protease and a minimal frustration principle is analyzed.

Inhibitory effect of 9-(2-phosphonylmethoxyethyl)adenine on visna virus infection in lambs: a model for in vivo testing of candidate anti-human immunodeficiency virus drugs.

The acyclic nucleoside phosphonate analog 9-(2-phosphonylmethoxyethyl)adenine (PMEA) was recently found to be effective as an inhibitor of visna virus replication and cytopathic effect in sheep choroid plexus cultures. To study whether PMEA also affects visna virus infection in sheep, two groups of four lambs each were inoculated intracerebrally with 10(6.3) TCID50 of visna virus strain KV1772 and treated subcutaneously three times a week with PMEA at 10 and 25 mg/kg, respectively. The treatment was begun on the day of virus inoculation and continued for 6 weeks. A group of four lambs were infected in the same way but were not treated. The lambs were bled weekly or biweekly and the leukocytes were tested for virus. At 7 weeks after infection, the animals were sacrificed, and cerebrospinal fluid (CSF) and samples of tissue from various areas of the brain and from lungs, spleen, and lymph nodes were collected for isolation of virus and for histopathologic examination. The PMEA treatment had a striking effect on visna virus infection, which was similar for both doses of the drug. Thus, the frequency of virus isolations was much lower in PMEA-treated than in untreated lambs. The difference was particularly pronounced in the blood, CSF, and brain tissue. Furthermore, CSF cell counts were much lower and inflammatory lesions in the brain were much less severe in the treated lambs than in the untreated controls. The results indicate that PMEA inhibits the propagation and spread of visna virus in infected lambs and prevents brain lesions, at least during early infection. The drug caused no noticeable side effects during the 6 weeks of treatment.

Stability and sensitivity analysis of Be-CoDiS, an epidemiological model to predict the spread of human diseases between countries. Validation with data from the 2014-16 West African Ebola Virus Disease epidemic.

Ebola virus disease is a lethal human and primate disease that requires a particular attention from the international health authorities due to important recent outbreaks in some Western African countries and isolated cases in European and North-America continents. Regarding the emergency of this situation, various decision tools, such as mathematical models, were developed to assist the authorities to focus their efforts in important factors to eradicate Ebola. In a previous work, we have proposed an original deterministic spatial-temporal model, called Be-CoDiS (Between-Countries Disease Spread), to study the evolution of human diseases within and between countries by taking into consideration the movement of people between geographical areas. This model was validated by considering numerical experiments regarding the 2014-16 West African Ebola Virus Disease epidemic. In this article, we propose to perform a stability analysis of Be-CoDiS. Our first objective is to study the equilibrium states of simplified versions of this model, limited to the cases of one an two countries, and to determine their basic reproduction ratios. Then, in order to give some recommendations for the allocation of resources used to control the disease, we perform a sensitivity analysis of those basic reproduction ratios regarding the model parameters. Finally, we validate the obtained results by considering numerical experiments based on data from the 2014-16 West African Ebola Virus Disease epidemic.

Field and depot maintenance manual : magneto, ignition, assembly, 2920-529-8247, 2920-740-3411, 2920-737-2995, and 2920-753-9854 (Bendix-Scintilla types S4LN-32 and S6LN-32), end item application, engine, gasoline, 4-cylinder, continental model AOI-268-3A and -3B (TM 9-7605-1) ....

"June 1960."

Operator's, organizational, direct support, and general support maintenance manual : pumping assembly, diesel engine driven, wheel mtd, 350 gpm, 275 ft. head, model 13220E1070 (97403) (fuel use only), NSN 4320-01-092-3551 ....

"15 August 1984."

Field and depot maintenance manual : engine, gasoline, fuel injected, supercharged, horizontal-opposed, air cooled, six cylinder, assembly-2805-528-6015, (Continental model AOSI-895-5 and AOSI-895-5M) : end item application : gun, self-propelled, full tracked, twin 40-mm, M42A1 (TM 9-2350-202), howitzer, self-propelled, full tracked, 105-mm, M52A1 (TM 9-2350-209) ....

Includes index.

Direct support and general support level : engine assembly, NSN 2815-00-050-8681, Detroit diesel model GM3-53.

Includes index.

Field and depot maintenance for engine, diesel (multifuel), turbosupercharged, fuel injected, water cooled, 6-cylinder, assembly-2815-897-5061, (Continental model LDS-427-2) and clutch, assembly (ORD 7748995), (Long model 13CF) : end item application, truck, cargo, 2 1/2 ton, 6 x 6, M35A1, (multifuel)--(TM 9-2320-235).

"February 1964."

Direct support and general support maintenance repair parts and special tools lists (including depot maintenance repair parts and special tools) : engine assembly, w/accessories (Continental model R-6602) and clutch (Rockford model 15TM) w/all accessories.

Includes index.

Direct and general support maintenance manual for transmission assembly, automatic, model TX 200-2A, Allison Division, GM, 2520-860-7342, transmission assembly, automatic, model TX 200-2B, Allison Division, GM, 2520-964-9207, transmission assembly, automatic, model TX 200-6, Allison Division, GM, 2520-045-3108.

Includes index.

Direct support and general support maintenance repair parts and special tools list ... transmission, crossdrive, hydromechanical, General Electric model HMPT 500 and transmission container assembly (2520-01-105-6446).

"January 1985."

Direct support and general support maintenance manual : engine, diesel, liquid cooled V-type, eight cylinder turbocharged, Cummins model VTA-903T with engine container assembly (2815-01-105-6455).

Includes index.

Direct support, general support, and depot maintenanc manual (including repair parts) : starter, engine, electrical, assembly 2920-999-6216 (Prestolite model--MFY6101AUT).

Includes indexes.