Structural identification of nonvolatile dimerization products of glucosamine by gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and nuclear magnetic resonance analysis

The degradation profile of glucosamine bulk form stressed at 100 degrees C for 2 h in an aqueous solution was studied. Column chromatography of acetylated product mixture led to isolation of two pure compounds (1b and 2b) and a mixture of at least three isomers (3b). 1a and 2a were identified as 5-(hydroxymethyl)-2-furaldehyde (5-HMF) and 2-(tetrahydroxybutyl)-5-(3',4'-dihydroxy-1'-trans-butenyl)pyrazine, respectively, by utilizing a variety of analytical techniques, such as GC-MS, LC-MS, on-line UV spectrum, (1)H and (13)C NMR, and DEPT, as well as (1)H-(1)H COSY. 3a was identified as 2-(tetrahydroxybutyl)-5-(2',3',4'-trihydroxybutyl)pyrazine, commonly known as deoxyfructosazine. In addition, glucosamine solid dosage form was exposed to 40 degrees C/75% relative humility for 10 weeks. Methanol extract of glucosamine solid dosage form was analyzed after acetylation by LC-MS, resulting in degradants 3b and 4b. 3a and 4a were, therefore, determined as deoxyfructosazine and 2,5-bis(tetrahydroxybutyl)pyrazine (fructosazine), respectively. Furthermore, the mechanisms of formation of identified degradation products are proposed and briefly discussed.     

Comparison of MALDI-TOF mass spectrometric to enzyme colorimetric quantification of glucose from enzyme-hydrolyzed starch

Successful quantification of the glucose produced by enzyme hydrolysis of starch was achieved by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protocol, using sorbitol as an internal standard. The starch contents measured by MALDI-TOF MS of corn starch, fiber-enriched oat flour derivatives, oat and barley flours, and barley flour/corn starch composites were evaluated in comparison to a widely accepted and validated method of starch determination, which relies on enzyme colorimetry (EC). The average starch content measured in a series of corn starch samples of different masses was 93 and 101% for EC and MALDI-TOF MS, respectively, values that represent the estimated purity of the sample. There was an agreement of 99% between the starch contents determined by the two analytical methods for complex flour-derived samples. Starch values estimated by MALDI-TOF MS consistently showed a greater degree of variability than those determined by EC, but this limitation was readily compensated by rapid acquisition of multiple mass spectra. This study is the first to report the quantification of glucose by MALDI-TOF MS, and it offers new perspectives into the potential utility of MALDI-TOF MS as a definitive tool for monosaccharide analysis and rapid starch determination in complex samples.     

Prednisone per os is likely to have limited efficacy in horses

Based on its efficacy for the treatment of human asthma, the corticosteroid prednisone is commonly used in horses for treatment of recurrent airway obstruction. However, recent studies have failed to show any benefit of prednisone tablets for the treatment of this condition. The purpose of this study was to determine why oral prednisone has poor efficacy for the treatment of heaves in horses. In a crossover study, 5 horses were given the following treatments: prednisone tablets, prednisone liquid, prednisolone tablets, prednisolone liquid and i.v. prednisolone sodium succinate (positive control). Blood samples were taken before drug administration and at selected time points during a 24 h period. Serum concentrations of prednisone and prednisolone were determined in order to evaluate gastrointestinal absorption and hepatic metabolism. Serum concentrations of the endogenous glucocorticoid hydrocortisone were also determined as an indicator of the biological activity of the drugs. Both prednisolone tablets and liquid were absorbed rapidly, with prednisolone detectable in serum within 15 min of administration and with peak concentrations occurring within 45 min. Small amounts of prednisone were detected in the serum samples after administration of both prednisone tablets and liquid. Prednisolone was not detected in serum samples after administration of prednisone liquid and was detected in serum samples from only one horse after administration of prednisone tablets. Endogenous hydrocortisone production was suppressed when horses received prednisolone. The results of these studies indicate that prednisone has poor efficacy for the treatment of heaves because it is poorly absorbed and the active metabolite prednisolone is rarely produced. In contrast, prednisolone tablets have excellent bioavailability and should be useful as a therapeutic agent in horses.     

Organophosphorus poisoning: A comparative study of the toxicity of carbophenothion to the Canada goose,the pigeon and the Japanese quail

The acute single dose oral toxicity of carbophenothion (S-4-chlorophenylthiomethyl OO-diethyl phosphorodithioate) has been determined in Canada geese (Branta canadensis), pigeons (Columba livia) and Japanese quail (Coturnix coturnix japonica). At post mortem examination gross pathological changes were observed in Canada geese and pigeons and esterase levels were determined by conventional and electrophoretic methods on extracts of liver and brain from these two species. Carbophenothion residue levels were determined in liver, brain and gizzard contents from the geese and pigeons. The overall pattern of results suggests that esterase inhibition may not be the dominant factor in carbophenothion poisoning in geese. It is suggested that a brain carbophenothion residue level of 1 part/10⁶ is indicative of death by poisoning in geese.     

Host immune responses after administration of inactivated Venezuelan equine encephalomyelitis virus vaccines—III. Kinetics for neutralizing antibody immunoglobulin class responses in donors and adoptively-immunized recipients

C57B16 mice were immunized with either live, attenuated TC-83 strain VEE virus vaccine or formalin-inactivated VEE vaccine combined with Bordetella pertussis. The kinetics of specific donor and adoptively-immunized recipient anti-VEE neutralizing antibody responses were studied. Donor mice immunized with either live or inactivated VEE virus vaccine combined with potent adjuvants develop specific anti-VEE IgM and IgG responses as early as 7 days post-immunization. Anti-VEE IgM antibody responses comprise the majority of anti-VEE neutralizing antibody at this early time period. By 14 to 21 days post-immunization, anti-VEE IgG responses predominated. When adoptively-immunized recipients were studied, the anti-VEE IgM to IgG predominance seen in donors early after administration was reversed, and for each time-period studied, recipients' serum anti-VEE antibody class responses consisted principally of IgG rather than IgM antibody. Since T-cells cooperation with B-cells is critical in the IgM-IgG antibody shift, these studies support the critical role T-cells exert in adoptive transfer in a murine model of experimental VEE infection. Furthermore, immunization with either live or inactivated VEE vaccine coupled to a potent adjuvant induce comparable donor and adoptively-immunized recipient anti-VEE antibody class responses.     

Impaired purine biosynthesis affects pathogenicity of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">melonis</Emphasis>

The vascular wilt pathogen Fusarium oxysporum f. sp. melonis causes worldwide yield losses of muskmelon. In this study, we characterized a UV-induced non-pathogenic mutant (strain 4/4) of F. oxysporum f. sp. melonis, previously identified as a potential biological control agent. During comparative analysis of vegetative growth parameters using different carbon sources, mutant strain 4/4 showed a delay in development and secretion of extracellular enzymes, compared to the wild type strain. Amendments of the growth medium with yeast extract, adenine or hypoxanthine, but not guanine, complemented the growth defect of strain 4/4, as well as secretion and partial activity of cellulases and endopolygalacturonases, indicating that the strain is an adenine auxotroph. Incubation of strain 4/4 conidia in adenine solution, prior to inoculation of muskmelon plants, partially restored pathogenicity to the mutant strain.     

Effect of surgical technique on limb function after surgery for rupture of the cranial cruciate ligament in dogs

OBJECTIVE: To determine the outcome and effect of surgical technique on limb function after surgery for rupture of the cranial cruciate ligament (RCCL) and injury to the medial meniscus in Labrador Retrievers. STUDY DESIGN: Prospective clinical study. ANIMALS: 131 Labrador Retrievers with unilateral RCCL and injury to the medial meniscus and 17 clinically normal Labrador Retrievers. PROCEDURE: Affected dogs had partial or complete medial meniscectomy and lateral suture stabilization (LSS), intracapsular stabilization (ICS), or tibial plateau leveling osteotomy (TPLO). Limb function was measured before surgery and 2 and 6 months after surgery. Treated dogs were evaluated to determine the probability that they could be differentiated from clinically normal dogs and tested to determine the likelihood that they achieved improvement. RESULTS: No difference was found between LSS or TPLO groups, but dogs treated with ICS had significantly lower ground reaction forces at 2 and 6 months. Compared with clinically normal dogs only, 14.9% of LSS-, 15% of ICS-, and 10.9% of TPLO-treated dogs had normal limb function. Improvement was seen in only 15% of dogs treated via ICS, 34% treated via TPLO, and 40% treated via LSS. CONCLUSIONS AND CLINICAL RELEVANCE: Surgical technique can influence limb function after surgery. Labrador Retrievers treated via LSS, ICS, or TPLO for repair for of RCCL and medial meniscal injury managed with partial or complete meniscectomy infrequently achieve normal function. Results of LSS and TPLO are similar and superior to ICS.     

Applied island biogeography and the conservation of endangered island birds in the Indian ocean

There has been much interest and controversy over the application of island biogeographic theory to conservation planning, but few attempts have been made actually to apply theory to specific conservation programmes. This paper demonstrates how island biogeography might play an important role in developing programmes to preserve several endangered island birds on the island of Mauritius. Biogeographic theory suggests that the present avifauna of Mauritius contains an excessive number of species, in view of the much reduced area of forest that exists on the island today. Inter-island transfers of four endangered birds from Mauritius to nearby Réunion Island are proposed and justified by biogeographical and ecological studies.     

Raman spectroscopic evidence for two conformations of uncomplexed valinomycin in the solid state

Raman spectroscopy is applied for the first time to elucidate the different conformations of the carrier transport molecule, valinomycin. Splitting of the ester and amide carbonyl stretch vibrations is observed in the Raman spectrum of crystals of valinomycin grown from both n-octane and acetone. These observations support the contention that some ester carbonyl groups are intramolecularly hydrogen bonded. The Raman spectrum of valinomycin grown from o-dichlorobenzene does not display this feature.     

In vitro inhibition of Eimeria tenella invasion by indigenous chicken Lactobacillus species

The aim of this study was to determine the effects of indigenous chicken Lactobacillus species isolates from different parts of the gastrointestinal tract on Eimeria tenella invasion in vitro and to characterise the nature of inhibition, if any. The effects of competitive exclusion, steric interference and bacterial extracellular factors on E. tenella invasion were examined in an MDBK cell model. Several Lactobacillus species were initially isolated from chickens and identified by biochemical characteristics and 16S-rRNA. All Lactobacillus species isolates tested, significantly inhibited E. tenella invasion. Steric interference did not affect parasite invasion. Extracellular metabolic factors secreted by Lactobacillus species isolates into the surrounding media were shown to inhibit parasite invasion and these factors appeared to be heat stable. These results show that the natural microflora of poultry can provide a source of E. tenella-inhibiting Lactobacillus species in vitro, and thus may contribute to the control of Eimeria infection.