D-glucosamine propanedithioacetal, an efficient chiral auxiliary in beta-lactam chemistry.

The synthesis of some monocyclic beta-lactams (monobactams) by the Staudinger reaction using D-glucosamine propanedithioacetal as chiral auxiliary is reported. The influence of several radicals at C3, C4, and C1' (sugar moiety) as well as other structural aspects are considered in relation to the antielastase activity.     

Morphological embedding and phonetic reduction: the case of triconstituent compounds

In this paper we propose that the internal bracketing of a word with more than two morphemes is reflected in the phonetic implementation. We hypothesize that embedded forms show more phonetic reduction than forms at higher structural levels (‘Embedded Reduction Hypothesis’). This paper tests the prediction of the Embedded Reduction Hypothesis with triconstituent compounds. The analysis of the durational properties of almost 500 compound tokens shows that there is a lengthening effect on the non-embedded constituent, and a shortening effect on the adjacent embedded constituent. Yet, this predicted effect of embedding interacts with other lexical factors, above all the bigram frequency of the embedded compound. At a theoretical level, these effects mean that the durational properties of the cross-boundary constituents are indicative of the hierarchical structure and of the strength of the internal boundary of triconstituent compounds. Hence, morphological structure is reflected in the speech signal.     

Desensitization, two-state receptors and pharmacological parameters

Katz' classical desensitization theory is examined and expanded. Two possible desensitization mechanisms are found to be thermodynamically identical with drug action on a two-state receptor and capable of interpretation as escape from an instability imposed by the activation of the receptor. The expanded theory of desensitization enables classical receptor theory to accomodate unexplained experimental facts such as the "fade" of both agonistic and antagonistic drug action and the metaphilic effect; it also makes it possible to interpret the variable efficacy or intrinsic activity of partial agonists as very rapid desensitization of drug-receptor complexes.     

The role of the pseudo-disaccharide neamine as an intermediate in the biosynthesis of neomycin.

By using wild-type and deoxystreptamine-negative mutants of Streptomyces fradiae grown in media containing [6(-3)H]glucose or [U-14C]glucose, and by subsequent hydrolysis of the labelled neomycin produced, neamines labelled with 3H in both rings I and II, but with 14C in ring I only, were prepared. A mixture of these two forms of neamine was converted by deoxystreptamine-negative Streptomyces rimosus forma paromomycinus into neomycin (not paromomycin) with a 30% yield. The3H: 14C ratio in this neomycin was the same as the measured in neamine produced by hydrolysis of the neomycin, and in unused neamine reisolated from the incubation medium. The 3H:14C ratio in the neomycin was not affected by the presence of unlabelled deoxystreptamine during the incubation. The radioactivity in the neomycin was associated with rings I and II only. It is concluded that the added neamine is incorporated into antibiotic intact, without initial hydrolysis, and that the probable first step in the subunit assembly of neomycin is the formation of neamine.     

Factors determining post-stimulation dilatation

Post-stimulation dilatation (PSD) of the femoral artery and vein after cessation of postganglionic sympathetic stimulation were related to the frequency and pulse number of the preceding stimulation. It was found that: 1) A minimum number of pulses (MNP) is needed to evoke PSD. MNP is inversely related to the stimulation frequency. A marked PSD develops after stimulation at 1 Hz when only 100 pulses were applied, whereas, if stimulated at 4 Hz or at higher frequencies, even 2,000 pulses fail to induce PSD. 2) The maximum value, the maximum rate and the overall diameter change of PSD (expressed either in absolute values or in relation to the preceding contraction) are a) directly related to the number of pulses at a constant stimulation frequency, b) for a constant number of pulses the above values are inversely related to the stimulation frequency. 3) The relation of PSD values to the stimulation parameters contradict the assumption that PSD is elicited either by a neurogenic transmitter released by the stimulation, or by an extraneuronal transmitter whose release is associated with the release of noradrenaline. PSD is suggested to be due to a decreased noradrenaline level within the synaptic cleft due to persistence of the reuptake after the release of noradrenaline had ceased.     

Development of allergic contact dermatitis requires activation of both tumor necrosis factor-receptors

We investigated the role of the TNF receptors, type I (p55TNFR) and type II (p75TNFR), in a mouse model of contact hypersensitivity, i.e., a model of a delayed type hypersensitivity (DTH) allergic reaction. Mice deficient for p55TNFR or p75TNFR were used to investigate the functions of these receptors in development of the DTH reaction. We show that both TNF receptors have a strong influence on the overall outcome of the DTH reaction, with the two TNF receptors exerting distinct functions. Dendritic cells of mice lacking p55TNFR had a defect in allergen uptake but showed normal migration into regional lymph nodes. In contrast, dendritic cells of p75TNFR-deficient mice showed diminished migration into regional lymph nodes after allergen contact, whereas the allergen uptake was independent of the p75TNFR. Thus, both TNF receptors are required for the development of a complete DTH reaction.     

Dynein supports motility of endoplasmic reticulum in the fungus Ustilago maydis

The endoplasmic reticulum (ER) of most vertebrate cells is spread out by kinesin-dependent transport along microtubules, whereas studies in Saccharomyces cerevisiae indicated that motility of fungal ER is an actin-based process. However, microtubules are of minor importance for organelle transport in yeast, but they are crucial for intracellular transport within numerous other fungi. Herein, we set out to elucidate the role of the tubulin cytoskeleton in ER organization and dynamics in the fungal pathogen Ustilago maydis. An ER-resident green fluorescent protein (GFP)-fusion protein localized to a peripheral network and the nuclear envelope. Tubules and patches within the network exhibited rapid dynein-driven motion along microtubules, whereas conventional kinesin did not participate in ER motility. Cortical ER organization was independent of microtubules or F-actin, but reformation of the network after experimental disruption was mediated by microtubules and dynein. In addition, a polar gradient of motile ER-GFP stained dots was detected that accumulated around the apical Golgi apparatus. Both the gradient and the Golgi apparatus were sensitive to brefeldin A or benomyl treatment, suggesting that the gradient represents microtubule-dependent vesicle trafficking between ER and Golgi. Our results demonstrate a role of cytoplasmic dynein and microtubules in motility, but not peripheral localization of the ER in U. maydis.     

Significant role for Fas in the pathogenesis of autoimmune diabetes

Programmed cell death represents an important pathogenic mechanism in various autoimmune diseases. Type I diabetes mellitus (IDDM) is a T cell-dependent autoimmune disease resulting in selective destruction of the beta cells of the islets of Langerhans. beta cell apoptosis has been associated with IDDM onset in both animal models and newly diagnosed diabetic patients. Several apoptotic pathways have been implicated in beta cell destruction, including Fas, perforin, and TNF-alpha. Evidence for Fas-mediated lysis of beta cells in the pathogenesis of IDDM in nonobese diabetic (NOD) mice includes: 1) Fas-deficient NOD mice bearing the lpr mutation (NOD-lpr/lpr) fail to develop IDDM; 2) transgenic expression of Fas ligand (FasL) on beta cells in NOD mice may result in accelerated IDDM; and 3) irradiated NOD-lpr/lpr mice are resistant to adoptive transfer of diabetes by cells from NOD mice. However, the interpretation of these results is complicated by the abnormal immune phenotype of NOD-lpr/lpr mice. Here we present novel evidence for the role of Fas/FasL interactions in the progression of NOD diabetes using two newly derived mouse strains. We show that NOD mice heterozygous for the FasL mutation gld, which have reduced functional FasL expression on T cells but no lymphadenopathy, fail to develop IDDM. Further, we show that NOD-lpr/lpr mice bearing the scid mutation (NOD-lpr/lpr-scid/scid), which eliminates the enhanced FasL-mediated lytic activity induced by Fas deficiency, still have delayed onset and reduced incidence of IDDM after adoptive transfer of diabetogenic NOD spleen cells. These results provide evidence that Fas/FasL-mediated programmed cell death plays a significant role in the pathogenesis of autoimmune diabetes.     

Chromosome aberrations in in vitro-produced bovine embryos at days 2-5 post-insemination

Availability of embryos of high quality is required to obtain satisfactory embryonic developmental rates and normal calves following transfer of in vitro-produced (IVP) bovine embryos. One relevant quality parameter is the frequency of chromosome aberrations, which can be evaluated using multicolor fluorescent in situ hybridization (FISH) with chromosome 6- and chromosome 7-specific probes in cattle. In this study, interphase nuclei (n = 3805) were analyzed from 426 bovine IVP embryos. We found that 73%, 72%, 81%, and 58% of the embryos from Days 2, 3, 4, and 5 post-insemination (pi), respectively, displayed a normal diploid chromosome number in all cells. When looking at the types of chromosome aberrations, the percentages of mixoploidy at Days 2, 3, 4, and 5 pi were 22%, 15%, 16%, and 42%, respectively, whereas the percentages of polyploidy (i.e., all nuclei in an embryo were analyzed and were polyploid) were 5%, 13%, 3%, and 0%, respectively. In conclusion, numerical chromosome aberrations were detected as early as Day 2 pi. The development of polyploid embryos is slow and is apparently arrested during the third cell cycle, whereas the mixoploid embryos seem to continue development.