Metabolism of bromhexine in pig hepatocyte cultures

The metabolism of bromhexine [N-cyclohexyl-N-methyl-2-(2-amino-3,5-di-bromo-benzyl)-amine] was studied using pig hepatocyte cultures and LC/MS/MS techniques. Phase I 'single-step' reactions, i.e. hydroxylation and demethylation occurred the fastest whereas the formation of hydroxylated/demethylated and aminal hydroxylated metabolites, which can be considered as multiple-step reactions, occurred more slowly. Phase II conjugates were detected for all hydroxylated metabolites. The glucuronides of the hydroxylated/demethylated components tended to accumulate. In addition to metabolites known to be formed in vivo, three unknown components related to bromhexine were detected. Two of these metabolites accumulated during incubation. Based on the fragmentation patterns, a possible molecular structure is proposed for these components.     

EX VIVO COMPUTED TOMOGRAPHIC EVALUATION OF MORPHOLOGY VARIATIONS IN EQUINE CERVICAL VERTEBRAE

Diagnostic imaging is one of the pillars in the clinical workup of horses with clinical signs of cervical spinal disease. An improved awareness of morphologic variations in equine cervical vertebrae would be helpful for interpreting findings. The aim of this anatomic study was to describe CT variations in left–right symmetry and morphology of the cervical and cervicothoracic vertebrae in a sample of horses. Postmortem CT examinations of the cervical spine for horses without congenital growth disorders were prospectively and retrospectively recruited. A total of 78 horses (27 foals, 51 mature horses) were evaluated. Twenty‐six horses (33.3%) had homologous changes in which a transposition of the caudal part of the transverse process (caudal ventral tubercle) of C6 toward the ventral aspect of the transverse process of C7 was present (n = 10 bilateral, n = 12 unilateral left‐sided, n = 4 unilateral right‐sided). There was one horse with occipito‐atlantal malformation, two horses with rudimentary first ribs bilaterally, and one horse with bilateral transverse processes at Th1, representing homeotic (transitional) vertebral changes. Chi‐square tests identified no significant differences in the number of conformational variations between the group of mature horses with or without clinical signs (P = 0.81) or between the group of mature horses and the group of foals (P = 0.72). Findings indicated that, in this sample of horses, the most frequently identified variations were homologous variations (transposition of the caudal part of the transverse process of C6–C7) in the caudal equine cervical vertebral column. Homeotic (transitional) variations at the cervicothoracic vertebral column were less common.     

Development of a biocatalytic process for the production of c6-aldehydes from vegetable oils by soybean lipoxygenase and recombinant hydroperoxide lyase

Volatile C6- and C9-aldehydes and alcohols are widely used as food flavors to reconstitute the "fresh green" odor of fruits and vegetables lost during processing. To meet the high demand for natural flavors, an efficient, cheap, and versatile biocatalytic process was developed to produce C6-aldehydes on a large scale. Vegetable oils were converted by soybean lipoxygenase and recombinant hydroperoxide lyase into hexanal and (2E)- or (3Z)-hexenal. In contrast to plant extracts, generally used as enzyme sources, high molar conversions were obtained with recombinant hydroperoxide lyase (50% for hexanal and 26% for hexenal formation), and no side products were formed. Furthermore, recombinant hydroperoxide lyase lacks isomerase activity, allowing production of (3Z)-hexenal, which could not be obtained in previously described processes. Recombinant hydroperoxide lyase is stable and can be stored at 4 degrees C for 1 month without significant loss of activity.     

The ecology of genetically modified mosquitoes

Ecological and population biology issues constitute serious challenges to the application of genetically modified mosquitos (GMM) for disease control.     

The inheritance of chemical phenotype in <Emphasis Type="Italic">Cannabis</Emphasis><Emphasis Type="Italic">sativa</Emphasis> L. (IV): cannabinoid-free plants

A genetic factor that blocks the cannabinoid biosynthesis in Cannabis sativa has been investigated. Crosses between cannabinoid-free material and high content, pharmaceutical clones were performed. F₁s were uniform and had cannabinoid contents much lower than the mean parental value. Inbred F₂ progenies segregated into discrete groups: a cannabinoid-free chemotype, a chemotype with relatively low cannabinoid content and one with relatively high content, in a monogenic 1:2:1 ratio. In our model the cannabinoid knockout factor is indicated as a recessive allele o, situated at locus O, which segregates independently from previously presented chemotype loci. The genotype o/o underlies the cannabinoid-free chemotype, O/o is expressed as an intermediate, low content chemotype, and O/O is the genotype of the high content chemotype. The data suggests that locus O governs a reaction in the pathway towards the phenolic cannabinoid precursors. The composition of terpenoids and various other compound classes of cannabinoid-free segregants remains unaffected. Backcrossing produced cannabinoid-free homologues of pharmaceutical production clones with potential applications in pharmacological research. A new variant of the previously presented allele ‘B ₀’, that almost completely obstructs the conversion of CBG into CBD, was also selected from the source population of the cannabinoid knockout factor.     

Variation in performance of western flower thrips populations on a susceptible and a partially resistant chrysanthemum cultivar

Variation in host plant performance among populations of a phytophagous insect pest is a potential threat to the durability of host plant resistance. Aggressive biotypes may overcome the protective properties of formerly resistant cultivars. Therefore, it is of interest to study such variation in breeding programs for host plant resistance to insects. In the present study, the performance of ten populations of Frankliniella occidentalis, the western flower thrips, was determined on a susceptible and a partially resistant cultivar of chrysanthemum, Dendranthema grandiflora. Damage, reproduction, and adult survival were determined using an excised leaf assay. Significant differences between the two cultivars and among the ten populations were found for all three characteristics. In general, damage, reproduction and adult survival were reduced on the resistant cultivar when compared to the susceptible cultivar. Some populations showed, in comparison to the reference population from the Netherlands, much higher damage and reproduction on one or both chrysanthemum cultivars. But also in these populations performance on the resistant cultivar was poor compared to the susceptible cultivar. This revised version was published online in August 2006 with corrections to the Cover Date.     

Granular envelope research in Pakistan

In 1989, research on granular envelope design for subsurface horizontal pipe drains was started when existing design procedures for envelope design did not result in successful application of a granular envelope (crushed rock) material at the Fourth Drainage Project. Although construction problems were, in part, the reason for the failure of the crushed rock envelope, it was also found that standard design criteria need additional rules to prevent a similar occurrence of failure. Tests were performed in the laboratory with permeameters, sieves, and other equipment and preliminary results are reported. The effect of number, size and arrangement of perforations on the test results are indicated. Test results of blended granular materials such as crushed rock, river-run gravel and sand from various sources in Pakistan are outlined.Proceedings of 5th International Drainage Workshop, Lahore, Pakistan, ICID-CIID, IWASRI, 1992, Vol. III, 5.38–5.49.     

Land-use related organic matter dynamics in North Cameroon soils assessed by 13C analysis of soil organic matter fractions

Topsoil samples from cultivated and adjacent non‐cultivated fields on three major agricultural soils in North Cameroon were fractionated into particle‐size fractions that were analysed subsequently for their C and ¹³C contents. The aim was to obtain further insight into the dynamics of soil organic matter (SOM) in relation to land use in Cameroon. Since organic carbon contents of the fractions were often very small, samples and analyses were extensively replicated to obtain robust statistical estimates of observed differences. For each soil type, differences in δ¹³C values between fields could be related to changes in the input and decomposition of organic matter arising from soil type, land management and, for example, the nature and abundance of weeds. Turnover of organic matter appeared to be fastest in the sand fraction, which is in line with results from earlier studies. In the finer fractions, clear differences in reaction to changes in input and decomposition were observed, that seem to be linked to differences in clay mineralogy. The results illustrate that SOM in the various fractions is much less stable and more strongly affected by changes in land use than might be assumed on the basis of changes in total SOM contents alone. At the same time, they demonstrate the relevance of ¹³C isotope analyses of SOM for studies on the impact of land use on these savannah soils with little SOM that are highly susceptible to degradation.