Antibody-induced internalization of viral glycoproteins in pseudorabies virus-infected monocytes and role of the cytoskeleton: a confocal study

Addition of pseudorabies virus (PrV)-specific polyclonal immunoglobulins to PrV-infected monocytes induces internalization of plasma membrane anchored viral glycoproteins. This process may interfere with antibody-dependent cell lysis and resembles the well-studied physiological endocytosis process. A confocal study was designed to investigate whether the major cellular components, involved in physiological endocytosis (clathrin, actin, dynein and microtubules), play a role in this virological internalization process. In order to visualize the interaction of endosomes, which contain the internalized viral glycoproteins, with clathrin, actin, dynein and microtubules, a double labeling of viral glycoproteins and different cellular proteins was performed. Porcine monocytes were inoculated with the PrV-strain 89V87 at a multiplicity of infection of 50 for 13h. After the addition of FITC-labeled porcine polyclonal PrV-specific antibodies, cells were fixed with para-formaldehyde at different time points and afterwards permeabilized. The different cellular components were visualized with monoclonal antibodies and a Texas Red-conjugate, with the exception of actin, which was stained with phalloidin-Texas Red. The cells were analyzed by confocal microscopy. A clear co-localization was observed between the viral glycoproteins and clathrin and dynein during the internalization process. The microtubules were in close contact with the internalized vesicles. For actin no co-localization could be observed. It can be stated that clathrin, dynein and microtubules, important components during physiological endocytosis, are also of importance during the antibody-induced internalization of viral glycoproteins.     

Geographical variation in water relations, hydraulic architecture and terpene composition of Aleppo pine seedlings from Italian provinces

Ecotypic variations in leaf conductance, soil-to-leaf hydraulic conductance, components of tissue water potential, hydraulic architecture parameters and xylem embolism were examined in greenhouse-grown two-year-old Aleppo pine (Pinus halepensis Mill.) seedlings from six origins representing the geographic range of the species in Italy. Cortical resin composition of the seedlings was also determined. Measurements were made on well-watered seedlings and on seedlings subjected to recurring severe drought. Drought-stressed seedlings had lower mean leaf conductances, transpiration rates and soil-to-leaf hydraulic conductances than well-watered seedlings. They also exhibited more negative osmotic potentials, higher relative water deficit at incipient plasmolysis, but a similar maximum modulus of elasticity. Drought-stressed seedlings showed a higher degree of xylem embolism, a lower Huber value, lower leaf specific conductivity and lower specific conductivity than well-watered seedlings. Drought-stressed seedlings of provenances from more xeric habitats (Tremiti, Porto Pino and Mottola) had greater leaf conductances, transpiration rates and soil-to-leaf hydraulic conductances than drought-stressed seedlings of provenances from more mesic habitats (Imperia, Otricoli and Vico del Gargano). They also showed higher osmotic adjustment and a lower degree of xylem embolism. Among provenances, there were no significant differences in hydraulic architecture parameters in response to the drought treatment; however, Tremiti and Porto Pino seedlings displayed smaller drought-induced reductions in specific conductivity and leaf specific conductivity, respectively, than seedlings from other provenances. These differences suggest that seedlings from xeric provenances, especially Tremiti, have greater resistance to desiccation than seedlings from mesic provenances. No clear association was found between terpene variability and the other traits investigated, although terpene composition was related to the geographical distribution of the provenances. We conclude that the drought-tolerance responses of Tremiti make it a more suitable provenance than the others for establishment on sites prone to severe soil water deficits.     

Self-Assembled Metal Colloid Monolayers: An Approach to SERS Substrates

The self-assembly of monodisperse gold and silver colloid particles into monolayers on polymer-coated substrates yields macroscopic surfaces that are highly active for surface-enhanced Raman scattering (SERS). Particles are bound to the substrate through multiple bonds between the colloidal metal and functional groups on the polymer such as cyanide (CN), amine (NH(2)), and thiol (SH). Surface evolution, which can be followed in real time by ultraviolet-visible spectroscopy and SERS, can be controlled to yield high reproducibility on both the nanometer and the centimeter scales. On conducting substrates, colloid monolayers are electrochemically addressable and behave like a collection of closely spaced microelectrodes. These favorable properties and the ease of monolayer construction suggest a widespread use for metal colloid-based substrates.     

Surgical correction of gastro‐oesophageal intussusception with bilateral incisional gastropexy in three dogs

Three dogs presented for evaluation of acute onset tachypnoea and dyspnoea following episodes of vomiting and/or regurgitation. Thoracic radiographs were suggestive of a gastro‐oesophageal intussusception in all three dogs; one dog also showed evidence of aspiration pneumonia. All three dogs underwent surgical correction with a bilateral incisional gastropexy. All dogs recovered from anaesthesia uneventfully and were discharged from the hospital 3 days after presentation. Persistent megaoesophagus was evident in all three dogs, and they are being chronically managed with a strict feeding regime and pro‐motility agents.     

Overberg Research Projects. XI. First stage larval reduction test to assess anthelmintic efficacy ante mortem in sheep.

Two field trials, one with suckling Merino ewe lambs and the other with yearling Dohne Merino rams, are described. In these the anthelmintic efficacy of febantel (a benzimidazole), ivermectin, levamisole and morantel are compared, using the 1st stage larval reduction test. The mean natural log (+1 for zero values) of the post treatment larval counts of the treated groups was compared with that of the untreated controls and the percentage reduction used to assess anthelmintic efficacy. Febantel was only 87.4% effective against Teladorsagia in suckling lambs but the other anthelmintics were more than 99% effective against this genus. Efficacy against Haemonchus and Trichostrongylus ranged from 93.2%-100% for all 4 compounds. In the rams all compounds were 100% effective against Trichostrongylus, with the exception of morantel which was only 87.5% effective. None of the compounds were effective against Teladorsagia, particularly morantel, animals treated with which having more larvae than the controls. The interpretation of anthelmintic efficacy; the advantages of the first stage larval reduction test, compared with the faecal egg count reduction test; and the importance of incubating cultures at 30 degrees C for 24 h, in order to harvest first stage larvae, are discussed.     

Composition and structure of the martian atmosphere: preliminary results from viking 1

Results from the aeroshell-mounted neutral mass spectrometer on Viking I indicate that the upper atmosphere of Mars is composed mainly of CO(2) with trace quantities of N(2), Ar, O, O(2), and CO. The mixing ratios by volume relative to CO(2) for N(2), Ar, and O(2) are about 0.06, 0.015, and 0.003, respectively, at an altitude near 135 kilometers. Molecular oxygen (O(2)(+)) is a major component of the ionosphere according to results from the retarding potential analyzer. The atmosphere between 140 and 200 kilometers has an average temperature of about 180 degrees +/- 20 degrees K. Atmospheric pressure at the landing site for Viking 1 was 7.3 millibars at an air temperature of 241 degrees K. The descent data are consistent with the view that CO(2) should be the major constituent of the lower martian atmosphere.     

MENSURATION OF THE NORMAL PITUITARY GLAND FROM MAGNETIC RESONANCE IMAGES IN 96 DOGS

The pituitary gland was measured from transverse magnetic resonance T1-weighted images after Gadolinium administration in 96 dogs weighing from 13 to 45 kg. The measurements were done by hand with calipers. The mean (+/- standard deviation) pituitary gland height was 5.1 mm (+/-0.9 mm). The mean width was 6.4 mm (+/- 1.1 mm). The correlation coefficient between pituitary and brain measurements, between pituitary measurement and body weight, and brain measurements and body weight was 0.0 to 0.3. A hyperintense region was present on T1-weighted images in the center of the pituitary gland in 64% of the dogs. At necropsy the pituitary glands were grossly and histologically normal. No pituitary gland measurements were performed at necropsy.     

Effects of Restored Stream Buffers on Water Quality in Non-tidal Streams in the Choptank River Basin

Restoration of riparian buffers is an important component of nutrient reduction strategies in the Chesapeake Bay watershed. In 1998, Maryland adopted a Conservation Reserve Enhancement Program (CREP), which provides financial incentives to take agricultural land out of production to plant streamside vegetation. Between 1998 and 2005, 1–30% of streamside vegetation (average?=?11%), was restored to forest or managed grass in 15 agriculturally dominated sub-basins in the Choptank River basin, a tributary of Chesapeake Bay. Pre-existing forested buffers represented 10–48% of the streamside (average?=?33%), for a total of 12–61% buffered streamsides (average?=?44%). Using multi-year water quality data collected before and after CREP implementation (1986, 2003–2006), we were unable to detect significant effects of CREP on baseflow nutrient concentrations based on the area of restored buffer, the percentage of restored streamside, or the percentage of total riparian buffer in the sub-basins (p?>?0.05). Although CREP increased the average buffered streamside from 33% in the 1990s to 44% by 2005, N and P concentrations have not changed or have increased in some streams over the last 20 years. Reductions may not have occurred for the following reasons: (1) buffer age, width, and connectivity (gaps) between buffers are also important to nutrient reductions; (2) agricultural nutrient inputs may have increased during this period; and (3) riparian buffer restoration was not extensive enough by 2005 to have measurable affects on the stream water quality in these sub-basins. Significant effects of CREP may yet be resolved as the current CREP buffers mature; however, water quality data through 2006 in the Choptank basin do not yet show any significant effects.     

Pharmacokinetics of chloramphenicol in calves during the first weeks of life

The pharmacokinetics of chloramphenicol, either administered as the monosuccinate ester or as a veterinary formulation, were studied in calves from the first day of life to the age of 10–12 weeks and compared with the results obtained in adult cattle. (1) After intravenous injection of 0.15 mmol/kg chloramphenicol monosuccinate, the plasma elimination half life of intact ester fell from a value of 33 min on the first day of life to 15 min at the age of 10–12 weeks (value in cows = 14 min). Free chloramphenicol reached maximal plasma concentrations after 2–3 h on the first day of life, but in less than 15 min in cows. The elimination half-life fell from about 15 h on day 1 to 4.8 h at the age of 10–12 weeks (4.2 h in cows). The bioavailability of the ester was more than 90% on Day 1, but declined to 50–60% from Day 7 on account of rapid renal excretion: 21–28% of the total dose was excreted as intact ester in a 2 h period following injection in calves aged 10–12 weeks. (2) The veterinary formulation of chloramphenicol proved toxic when administered intravenously at a dose of 0.15 mmol/kg, and even a dose of 0.093 mmol/kg was less well tolerated than 0.15 mmol/kg of the monosuccinate ester. (3) The pharmacokinetics of chloramphenicol fitted an open two-compartment model, the half-life of the elimination phase corresponded well to the values determined in the experiments with the monosuccinate ester. (4) The intramuscular injection of 0.15 mmol/kg of the ester or 0.093 mmol/kg of chloramphenicol provided ‘therapeutic’ plasma concentrations (≥ 5 μg/ml) within 15–30 min and for about 24 h in calves aged 7 days. (5) Chloramphenicol crossed the placenta when given to cows shortly before a Caesarian section, but equilibrium was not reached within 50–100 min. (6) The binding of chloramphenicol to serum proteins was dependent both on total protein and drug concentrations. It rose from less than 30% on day 1 to about 40% in adult cattle. (7) Recommendations for a dosage regime for chloramphenicol in calves are made on the basis of the pharmacokinetic data.