Synthesis of novel paclitaxel prodrugs designed for bioreductive activation in hypoxic tumour tissue.

The syntheses and preliminary evaluation of the first potential bioreductive paclitaxel prodrugs are described. These prodrugs were designed as potential candidates in more selective chemotherapy by targeting hypoxic tumour tissue. Aromatic nitro and azide groups were used as the bioreductive trigger. Generation of paclitaxel occurs after reduction and subsequent 1,6-elimination or 1,8-elimination. All prodrugs are stable in buffer and indeed give paclitaxel after chemical reduction of the aromatic nitro or azide functionality. In aerobic cytotoxicity assays several prodrugs exhibit diminished cytotoxicity. These compounds are interesting candidates for further biological evaluation.     

A lamB expression plasmid for extending the host range of phage λ to other enterobacteria

Abstract We have constructed a multicopy plasmid vector (pAMH62) expressing lamB , the gene coding for the phage λ receptor protein in Escherichia coli . In this construction, the lamB structural gene was fused to the ompR promoter of E. coli . The ompR promoter was employed because: (i) it can function in other gram negative bacteria; (ii) it expresses lamB in a multicopy state at a level comparable to that of maltose-induced chromosomal lamB in E. coli . The vector pAMH62 was tested in E. coli and Salmonella typhimurium . In both cases the LamB protein was produced in similar amounts, was properly integrated to the outer membrane and was functional as phage λ receptor. Thus pAMH62 should provide a useful tool for extending the host range of phage λ and λ-derived vectors to other Gram-negative bacteria.     

Relationship between local perfusion and FFA uptake in human skeletal muscle-no effect of increased physical activity and aerobic fitness.

We investigated heredity-independent effects of increased physical activity and aerobic fitness on skeletal muscle free fatty acid (FFA) uptake, perfusion, and their heterogeneity at rest and during exercise. Also, the relationship between local skeletal muscle FFA uptake and perfusion was studied. Nine young adult male monozygotic twin pairs with significant difference in physical activity [229 min (SD 156) average time spent for conditioning exercise per week in more and 98 min (SD 71) in less active twins, P = 0.013] and aerobic fitness [18% (SD 10) difference in maximum O2 uptake] between brothers were studied using positron emission tomography. Submaximal knee-extension exercise increased perfusion, FFA uptake, and oxygen uptake in quadriceps femoris muscles 6-10 times compared with resting values (P < 0.001). More active twins tended to utilize more oxygen, while no differences were found in muscle perfusion or FFA uptake between groups. Mean perfusion and FFA uptake correlated strongly at a whole muscle level, both at rest (r = 0.97, P = 0.03 in more and r = 0.98, P = 0.02 in less active twins) and during exercise (r = 0.99, P = 0.01 and r = 0.94, P = 0.06), but at the voxel level (87 mm3) correlation was only moderate during exercise [r = 0.73 (SD 0.08) vs. r = 0.74 (SD 0.10), P = 0.92] and weak at rest [r = 0.28 (SD 0.13) vs. r = 0.33 (SD 0.21), P = 0.58]. Exercise decreased both perfusion and FFA uptake heterogeneity within the muscles (P < 0.001) similarly in both groups. In conclusion, long-term history of moderately increased physical activity tends to enhance muscle oxidative metabolism, but it does not have any significant influence on the FFA uptake or perfusion rates or their heterogeneity in skeletal muscle. Submaximal knee-extension exercise decreases heterogeneity of muscle FFA uptake and perfusion and improves matching between local muscle perfusion and FFA uptake. Thus it seems that the genetic influence is more important to determine the heterogeneity of perfusion and FFA uptake in skeletal muscle than exercise training.     

Screening of free 17-alkyl-substituted anabolic steroids in human urine by liquid chromatography-electrospray ionization tandem mass spectrometry

A qualitative liquid chromatography-electrospray ionization tandem mass spectrometry method was developed for screening of the abuse of 4-chlorodehydromethyltestosterone, danazol, fluoxymesterone, formebolone, metandienone, oxandrolone, and stanozolol. The introduced method measures simultaneously nine different 17-alkyl-substituted anabolic androgenic steroids or their unconjugated metabolites in human urine, using methyltestosterone as an internal standard. Sample preparation involved one-step liquid extraction. Liquid chromatographic separation was achieved on a reversed-phase column with methanol-water gradient containing 5 mmol/l ammonium acetate and 0.01% (v/v) acetic acid. Compounds were ionized in the positive mode and detected by multiple reaction monitoring. All steroids within the study could be selectively detected in urine with detection limits of 0.1-2.0 ng/ml. The method showed good linearity up to 250 ng/ml with correlation coefficients higher than 0.9947. With simple and fast sample preparation, low limits of detection, and high selectivity and precision, the developed method provides advantages over the present testing methods and has the potential for routine qualitative screening method of unconjugated 17-alkyl-substituted anabolic steroids in human urine.     

Bacteriological Quality of Raw Materials Used in Finnish Mink Feed

Mink feed raw materials were analyzed for total bacterial count, the number of faecal streptococci, the coliform count, the number of haemolytic bacteria and the number of sulphite reducing bacteria. The investigation comprised samples from the following raw materials: four slaughter-house offal products, preserved and unpreserved slaughter blood, Baltic herring, cod filletting offal, fish silage, blood meal, fish meal, meat-bone meal, protein concentrate, brewer’s yeast and cereal feed. The slaughter-house offals and unpreserved slaughter blood had the poorest quality, in terms of all the bacterial types for which the samples were analyzed. There were statistically significant differences in bacterial contents between slaughter-house offals from different sources. The preserved slaughter blood had significantly lower bacterial contents as compared to the unpreserved slaughter blood. Single samples of the cod filletting offal, Baltic herring and the blood meal had relatively high total bacterial counts, but the specified mean bacterial counts were relatively low. The bacterial counts for the rest of the investigated raw materials were relatively low.     

Altering glucosinolate profiles modulates disease resistance in plants

Plant diseases are major contributing factors for crop loss in agriculture. Here, we show that Arabidopsis plants with high levels of novel glucosinolates (GSs) as a result of the introduction of single CYP79 genes exhibit altered disease resistance. Arabidopsis expressing CYP79D2 from cassava accumulated aliphatic isopropyl and methylpropyl GS, and showed enhanced resistance against the bacterial soft-rot pathogen Erwinia carotovora, whereas Arabidopsis expressing the sorghum CYP79A1 or over-expressing the endogenous CYP79A2 accumulated p-hydroxybenzyl or benzyl GS, respectively, and showed increased resistance towards the bacterial pathogen Pseudomonas syringae. In addition to the direct toxic effects of GS breakdown products, increased accumulation of aromatic GSs was shown to stimulate salicylic acid-mediated defenses while suppressing jasmonate-dependent defenses, as manifested in enhanced susceptibility to the fungus Alternaria brassicicola. Arabidopsis with modified GS profiles provide important tools for evaluating the biological effects of individual GSs and thereby show potential as biotechnological tools for the generation of plants with tailor-made disease resistance.     

A short-lived herbivore on a long-lived host: tree resistance to herbivory depends on leaf age

Short-lived insect herbivores should be able to adapt to the resistance mechanisms of their long-lived woody hosts because the life span of a single host will encompass numerous generations of herbivores. However, adaptation may be slowed down if host genotypes can create, in a single genotype, such large phenotypic variation in traits relevant for the herbivore that it matches variance among host genotypes. We tested this hypothesis by measuring leaf consumption by, and growth of, half-sibs of the geometrid moth Epirrita autumnata on individual birch trees, during three instars. The instar×tree interaction, rather than tree identity alone, was a significant variance component for both consumption and growth, indicating that different larval instars ranked individual trees differently. Both consumption and growth varied most between the 3rd and the later (4th and 5th) instars, coinciding with rapid seasonal changes in numerous nutritive and phenolic traits of maturing leaves. Thus, developmental variance in the leaf quality of individual trees may reduce the likelihood of E. autumnata genotypes adapting to the defenses of their host trees. We did not find evidence of in the ability of different half-sibs to utilize individual trees or leaf stages, indicating that E. autumnata larvae are generalists over a wide variety of host traits.