Impact of ubiquitous inhibitors on the GUS gene reporter system: evidence from the model plants Arabidopsis, tobacco and rice and correction methods for quantitative assays of transgenic and endogenous GUS

Background  

The β-glucuronidase (GUS) gene reporter system is one of the most effective and employed techniques in the study of gene regulation in plant molecular biology. Improving protocols for GUS assays have rendered the original method described by Jefferson amenable to various requirements and conditions, but the serious limitation caused by inhibitors of the enzyme activity in plant tissues has thus far been underestimated.     

Purification of xylitol obtained by fermentation of corncob hydrolysates

Hydrolysates obtained by autohydrolysis-posthydrolysis of corncobs were detoxified with charcoal, concentrated, supplemented with nutrients, and fermented with Debaryomyces hansenii. After biomass removal, the fermented media contained 0.1137 kg of nonvolatile components (NVC)/kg of liquor, which corresponded mainly to xylitol (0.6249 kg/kg of NVC) but also to minor amounts of inorganic components (measured as ashes), proteins, nonfermented sugars (xylose and arabinose), uronic acids, arabitol, and other nonvolatile components (ONVC). The media were subjected to further processing (sequential stages of adsorption, concentration, ethanol precipitation, concentration, and crystallization) to obtain food-grade xylitol. Adsorption experiments were carried out at various solid-to-liquor ratios. Under selected conditions (1 kg of charcoal/15 kg of liquors), the xylitol content increased to 0.6873 kg/kg of NVC, and almost total decoloration was achieved. The resulting liquor was concentrated by evaporation to increase its NVC content to 0.4032 kg/kg of liquor (corresponding to a xylitol concentration of 0.280 kg/kg of liquor), and ethanol was added to precipitate a part of the NVC (mainly proteins, but also uronic acids, ashes, and other nonvolatile compounds). Refined liquors (containing 0.7303 kg of xylitol/kg of NVC) were concentrated again, and ethanol was added (to reach 40-60% volume of the stream) to allow crystallization at -10 or -5 degrees C. Under selected conditions, 43.7% of xylitol contained in the initial fermentation broth was recovered in well-formed, homogeneous crystals, in which xylitol accounted for 98.9% of the total oven-dry weight. Material balances are presented for the whole processing scheme considered in this work.     

Assessment of EST- and genomic microsatellite markers for variety discrimination and genetic diversity studies in wheat

It is likely that in the near future sequence information from sequencing programmes and EST libraries will generate an abundance of genic microsatellite markers. This study is focused on the assessment of their likely impact and performance vis-à-vis their genomic counterparts. Microsatellites from two sources were used to assess the genetic diversity in 56 old and new varieties of bread wheat on the UK Recommended List. A set of 12 microsatellite markers generated from genomic libraries and 20 expressed sequence tag (EST)-derived microsatellites were used in the study, and the performance of both marker sets assessed. The EST-derived or genic microsatellites delivered fingerprints of superior quality, amplifying clear products with few stutter bands. Diversity levels as revealed bygenic microsatellites are similar to the few published results. The PIC values for the genic markers were generally lower than those calculated for the genomic microsatellites, though advantages of both marker classes for variety identification applications are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

The impact of MK-467 on sedation,heart rate and arterial blood pressure after intramuscular coadministration with dexmedetomidine in conscious cats

Objective

To study the effects of MK-467, a peripheral α₂-adrenoceptor antagonist, on sedation, heart rate and blood pressure after intramuscular (IM) coadministration with 25 μg kg^?1 of dexmedetomidine in cats.

Renal diseases of reptiles.

Renal disease is one of the most common problems in reptilian medicine. Herbivorous species seem to be more susceptible than carnivores. The causation may be multifactorial, and the signs are often nonspecific. Prevention is preferred to treatment, and a comprehensive health program, which includes quarantine, routine health screens during annual examinations, and necropsy examinations of all animals that die, is desirable.     

EXCRETORY UROGRAPHY BY INTRAOSSEOUS INJECTION OF CONTRAST MEDIA IN A RABBIT MODEL

There are many indications for an intravenous excretory urogram. However, where intravenous access is not available, the intraosseous route to the circulation may be an alternative. We found that safe and diagnostic excretory urograms could be obtained in rabbits following the injection of different contrast media via the intraosseous route. In fact, these excretory urograms were indistinguishable from ones obtained by the conventional intravenous route. While the rabbits did not develop any abnormal clinical signs following the procedure, there were postmortem histologic lesions of osteochondrosis in 5 of 22 (22.7%) tibias receiving an intraosseous needle, but in none of the 14 tibias that did not receive an intraosseous needle. Further, the use of diatrizoate was associated with the development of osteochondrosis while the use of iopamidol was not.     

Effects of epidurally administered morphine or buprenorphine on the thermal threshold in cats

Objective: To determine the antinociceptive effects of epidural administration of morphine or buprenorphine in cats by use of a thermal threshold model. ANIMALS: 6 healthy adult cats. PROCEDURES: Baseline thermal threshold was determined in duplicate. Cats were anesthetized with isoflurane in oxygen. Morphine (100 microg/kg diluted with saline [0.9% NaCl] solution to a total volume of 0.3 mL/kg), buprenorphine (12.5 microg/kg diluted with saline solution to a total volume of 0.3 mL/kg), or saline solution (0.3 mL/kg) was administered into the epidural space according to a Latin square design. Thermal threshold was determined at various times up to 24 hours after epidural injection. RESULTS: Epidural administration of saline solution did not affect thermal threshold. Thermal threshold was significantly higher after epidural administration of morphine and buprenorphine, compared with the effect of saline solution, from 1 to 16 hours and 1 to 10 hours, respectively. Maximum (cutout) temperature was reached without the cat reacting in 0, 74, and 11 occasions in the saline solution, morphine, and buprenorphine groups, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Epidural administration of morphine and buprenorphine induced thermal antinociception in cats. At the doses used in this study, the effect of morphine lasted longer and was more intense than that of buprenorphine.     

Skull size and shape variation in Psammomys spp. (Rodentia,Gerbillinae) from Tunisia,with emphasis on the impact of allometric variation on species recognition

In Tunisia both Psammomys obesus and P. vexillaris are found. These taxa have been the subject of taxonomic controversy for some time, due to variability in the classical morphological characters used for taxonomic recognition. In this study we investigated skull size and shape variation in the genus Psammomys by using geometric morphometrics to evaluate the extent of intra- and interspecific shape variation and explicitly tested for the impact of allometric shape variation on species discrimination. Eleven populations of the two species from 10 localities in Tunisia were studied. Statistical analyses of size and shape showed large size variation within P. obesus, but no shape differences were revealed among populations of this species. Interspecific analysis revealed that P. vexillaris had the smallest skull. Principal component analysis and Procrustes distances showed good discrimination between the two species after removal of the allometric component of shape variation. The results obtained show that allometric-related shape variation could mask discrimination between the two Psammomys species. This finding might explain the uncertainty in classification of these species in the past. The interspecific allometric-free phenotypic differences observed may be associated with adaptive processes linked to the different environmental and trophic preferences of the two species.     

Specific detection of Histomonas meleagridis in turkeys by a PCR assay with an internal amplification control

Histomoniasis or blackhead is a disease of gallinaceous birds, caused by the protozoon Histomonas meleagridis. Since traditional diagnostics for the detection of this disease are complex and far less sensitive than molecular tools, a PCR would provide a more rapid and sensitive alternative. However, intestinal material and droppings, which are preferably used in epidemiological studies of histomoniasis, often contain PCR inhibitory substances. To detect these false negative results, the use of an internal amplification control is essential. Nevertheless, the recently developed PCR tests lack this internal control. Therefore, a new PCR assay with H. meleagridis specific primers was developed which does include an internal amplification control. The diagnostic value of the PCR assay was evaluated in comparison to three other conventional H. meleagridis specific PCR tests (HIS5, HM1 and HM2). None of the organ samples originating from uninfected turkeys, showed positive PCR results in any of the tests. Among the lesion-positive, inhibition-free samples, 95.4% were positive by our PCR assay, while only 50, 66.7 and 83.3% of the lesion-positive organs tested positive by the HM1, the HIS5 and the HM2 PCR respectively. In conclusion, our PCR offers the use of the internal control to detect false negative results and an increased sensitivity, and thus should be useful for routine diagnosis of H. meleagridis in poultry.