Combinatorial approach to flavor analysis. 1. Preparation and characterization of a S-methyl thioester library.

A new method for the "one-pot" synthesis of S-methyl thioesters has been developed by reacting methyl chlorothiolformate with carboxylic acids. The resulting "flavor library" contained all the intended thioesters and a single major impurity, identified by GC-MS as S, S-dimethyldithiocarbonate. Quantification of individual compounds present in the library was performed by GC analysis using two independent methods of detection, SCD and FID. It was shown that apart from S-methyl thioacetate (0.8 mol %), molar concentrations of other thioesters varied in a relatively narrow range from 4.2 mol % for S-methyl thiopropionate to 14.1 mol % for S-methyl thiohexanoate. In general, medium chain S-methyl thioesters were present in slightly higher molar concentrations than those prepared from short or long chain carboxylic acids. This variation was attributed to partial loss of the most volatile components during extraction and the lower reactivity of higher homologues. The library was used for the characterization of some physicochemical parameters of thioesters. In particular, lipophilicity coefficients (log k(w)) and thioester retention in 10, 20, and 33% triolein (used as a model lipid phase) were determined directly by reverse-phase HPLC and extrapolated from the respective data. This analysis illustrates that substantial information can be generated using a library containing a relatively large number of compounds in effectively the same way as is necessary for the analysis of a single sample.     

Occurrence of odorant polyfunctional thiols in the Super Alpha Tomahawk hop cultivar. Comparison with the thiol-rich Nelson Sauvin bitter variety

Tomahawk hop (Humulus lupulus) is a recently developed Super Alpha cultivar (14-18% α-acids w/w), already widely used by brewers to impart bitterness and a citrus-like aroma to beer. By comparison with two bitter varieties (Nelson Sauvin and Nugget) and two aromatic ones (Cascade and Saaz), the Tomahawk cultivar showed a very particular terpenoid profile, rich in both α- and β-selinenes (>600 mg/kg IST equiv in total), methyl geranate (>40 mg/kg IST equiv), and geraniol (>200 mg/kg). Tomahawk also proved to contain a wide variety of odorant polyfunctional thiols. The major β-sulfanyl acetate, 3-sulfanyl-2-ethylpropyl acetate, newly identified here, was found at similar levels in the famous Sauvignon-like Nelson Sauvin and Tomahawk varieties (15-44 μg/kg IST equiv). On the other hand, lower levels of total β-sulfanyl alcohols were measured in Tomahawk, although 3-sulfanylhexan-1-ol was found at a similar level and the 3-sulfanyl-4-methylpentan-1-ol previously claimed to be specific to the Nelson Sauvin variety was also evidenced in the Super Alpha cultivar (9-13 μg/kg IST equiv). As revealed by boiling and fermentation, Tomahawk hop also contains very interesting bound polyfunctional thiols that should be investigated for better use by brewers.     

Early enhancement of calcium currents by H-ras oncoproteins injected into Hermissenda neurons

Influx of calcium through membrane channels is an important initial step in signal transduction of growth signals. Therefore, the effects of Ras protein injection on calcium currents across the soma membrane of an identified neuron of the snail Hermissenda were examined. With the use of these post-mitotic cells, a voltage-sensitive, inward calcium current was increased 10 to 20 minutes after Harvey-ras oncoproteins were injected. The effects of oncogenic Harvey ras p21 protein (v-Ras) occurred quickly and were sustained, whereas the effects of proto-oncogenic ras protein (c-Ras) were transient. This relative potency is consistent with the activities of these oncoproteins in stimulating cell proliferation. Thus, this calcium channel may be a target for Ras action.     

A conserved mutation in an ethylene biosynthesis enzyme leads to andromonoecy in melons

Andromonoecy is a widespread sexual system in angiosperms characterized by plants carrying both male and bisexual flowers. In melon, this sexual form is controlled by the identity of the alleles at the andromonoecious (a) locus. Cloning of the a gene reveals that andromonoecy results from a mutation in the active site of 1-aminocyclopropane-1-carboxylic acid synthase. Expression of the active enzyme inhibits the development of the male organs and is not required for carpel development. A causal single-nucleotide polymorphism associated with andromonoecy was identified, which suggests that the a allele has been under recent positive selection and may be linked to the evolution of this sexual system.     

Suspected fatal venous air embolism during anaesthesia in a Pomeranian dog with pulmonary calcification

CASE HISTORY: Death occurred in a 1.25 kg, 9-month-old female Pomeranian dog undergoing anaesthesia for surgical repair of partially healed fractures of the radius and ulna.

CLINICAL FINDINGS: Following sedation, anaesthesia was induced using thiopentone and maintained with halothane in oxygen. An acute decrease in the dog's end-tidal carbon dioxide (EtCO₂) measurements was noted approximately 50 min after induction, immediately following delivery of a 5-ml bolus of lactated Ringer's solution (LRS) administered to flush a small (0.06 ml) volume of fentanyl via a pre-placed intravenous (I/V) catheter. Venous air embolism (VAE) was suspected and the dog died despite interventive therapy. On post-mortem examination, several air bubbles were noted when the right ventricle was opened under water. Histologically, the kidneys appeared abnormal with immature glomeruli, and the lungs appeared diffusely mineralised. The origin of the air was probably small bubbles and microbubbles that may have been present in the extension set and 20 ml syringe used for the administration of fentanyl and I/V fluids to the dog.

DIAGNOSIS: Renal dysplasia and diffuse pulmonary calcification, with VAE as the probable cause of death.

CLINICAL RELEVANCE: In this case of VAE-associated anaesthetic death, it is further speculated that underlying pulmonary disease, in the form of pulmonary calcification, may have contributed to an increased sensitivity to the adverse effects of VAE.     

Emerging technologies for identifying superior dairy cows in New Zealand

The performance of animals is determined by the interaction of their genes with environmental circumstances. Accordingly, animals exhibiting superior performance are not necessarily the animals with the best genes nor are they the best choice of parents. Statistical analyses of production records for repeated traits, e.g. lactation yields and reproductive performance, show that part of the variation in performance among animals in the same herd and year is due to genetic differences, and the remainder is due to so-called residual or environmental factors that are not passed on to offspring. These within-herd environmental factors can be partitioned into a component that affects performance throughout an animal's lifetime, and a part that is unique to each observation. The process of animal evaluation from pedigree and performance records partitions the superiority of each cow into these three components. Reliable assessment of the genetic merit of bulls has required progeny testing, and for cows has required observation of their own individual performance. Selection on the genetic or breeding value component has systematically improved animal performance over recent decades, but has been limited by the age at which assessments of genetic merit are available.

Emerging molecular technologies can read DNA sequences or measure RNA expression and have allowed the identification of a number of chromosome regions, and a few specific genes in those regions, that influence economic performance. This information allows better characterisation of the relationships between animals and more accurate predictions of genetic merit in bulls without progeny information and in cows that have yet to produce their own performance record. At some stage, enough genes responsible for variation in performance will be identified to allow faster genetic progress through selection of animals at young ages and therefore more rapid turnover of the generations. Mechanisms that modify gene expression have been identified and these may ultimately allow animals to be selected at an early age for lifetime productivity, accounting for processes that modify gene expression and lead to differences in performance that are not reflected by DNA sequence information. This review describes the status of these emerging technologies and their likely role in the improvement of dairy cattle.     

Abortions in sheep caused by Salmonella Brandenburg: Pathological findings

CASE HISTORY: A 7-year-old cat developed sporadic vomiting, reduced appetite, and weight loss over the previous 3 months.

CLINICAL FINDINGS: Palpation revealed a large mid-abdominal mass and the cat had marked eosinophilia. The cat progressively lost weight over the next 7 weeks when euthanasia was performed.

PATHOLOGICAL FINDINGS: Necropsy revealed a 3?cm diameter firm white intramural mass in the colon and another in the pylorus. Mesenteric and cranial mediastinal lymph nodes were firm, pale, and enlarged. Histopathological examination revealed foci of necrosis surrounded by thick dense collagen trabeculae and predominantly eosinophilic inflammation within the intestine and lymph nodes. Marked eosinophilic infiltration of the liver was also present.

DIAGNOSIS: The lesions were consistent with gastrointestinal eosinophilic sclerosing fibroplasia (FGESF).

CLINICAL RELEVANCE: This is the first report of FGESF in a New Zealand cat and the first time lesions of FGESF have been observed in extra-abdominal tissues. Intestinal neoplasia can be clinically identical to FGESF and histopathology is required for differentiation. Evidence suggests that FGESF has a more favourable prognosis than intestinal neoplasia.     

Salmonella Brandenburg — emergence of a variant strain on a sheep farm in the South Island of New Zealand

AIM: To compare the rectal and I/V administration of tramadol in dogs, to assess both its pharmacokinetic properties and absolute bioavailability.

METHODS: After rectal administration via suppositories and I/V injection of tramadol (4 mg/kg), the concentration of tramadol and its main metabolites, O-desmethyl-tramadol (M1), N-desmethyl-tramadol (M2) and N,O-didesmethyl-tramadol (M5), were determined in plasma, using high-performance liquid chromatography (HPLC). A balanced cross-over study was used, involving six male Beagle dogs.

RESULTS: Plasma concentrations after rectal and I/V administration were fitted on the basis of mono- and bi-compartmental models, respectively. Following rectal administration tramadol was detected from 5 minutes up to 10 hours, in lesser amounts than M5 and M2, while M1 was detected in negligible amounts. Following I/V administration tramadol was detected up to 10 hours, M2 and M5 were detected at similar concentrations, and M1 was present at low concentrations. The area under the curve (AUC) of the three metabolites did not differ significantly after either route of administration of tramadol. The absolute bioavailability of tramadol via rectal administration was 10 (SD 4)%.

CONCLUSIONS: After rectal administration of tramadol suppositories, absorption of the active ingredient was rapid, but its metabolism quickly transformed the parent drug to high levels of M2 and M5.

CLINICAL RELEVANCE: In the dog, rectal pharmaceutical formulation of tramadol would have a different pharmacokinetic behaviour than in humans.     

In vivo selection for salt tolerance in Coleus blumei

The aim of the investigation was to examine by in vivo screening the potential for salt tolerance in a selected cultivar of the amenity annual, Coleus blumei. Seedlings were placed on a capillary mat soaked in salinized nutrient solution. An initial dose response over the range 0-120 mM NaCl showed that at 90 mM NaCl only 15% of seedlings survived. A large scale screen of seedlings at 90 mM produced a number of survivors of which about 2% grew normally. Cuttings from these survivors showed more tolerance than cuttings from unselected plants. Seed progeny from some of these surviving plants were vegetatively more vigorous in the presence of 90 mM NaCl than seed progeny from unselected plants. Using this simple technique it was possible to screen large numbers of seedlings for salt tolerance in this species. It is suggested that the technique could also be used to screen for a wide range of other characters.