Ru-catalysed C-H silylation of unprotected gramines, tryptamines and their congeners.

Selective Ru-catalysed C2-H silylation of heteroarenes is presented. The transformation works with or without directing group assistance and requires no protecting groups. Gramines and tryptamines may be converted efficiently whilst avoiding deleterious elimination side-reactions. Mechanistic studies reveal an unusual activation of the indole C4-H bond by an electron-rich metal.

Electrospray ionisation Fourier-transform ion cyclotron resonance mass spectrometry of dynamic combinatorial libraries.

The use of electrospray ionisation Fourier-transform ion cyclotron resonance tandem mass spectrometry (ESI-FTICR-MS/MS) for the analysis of dynamic combinatorial libraries (DCLs) of pseudo-peptide macrocyclic hydrazone oligomers is presented. The design of library building blocks results in mixtures of compounds with greater diversity than libraries generated by conventional combinatorial chemistry and so presents increased demands for analysis. The extended capabilities of the FTICR technique, specifically selective ion trapping, sensitivity, high resolution and mass accuracy over a broad mass range, are compatible with these increased demands and, most importantly, without the need for chromatography. Preliminary studies on the sequencing of cyclic oligomers and confirmation of the presence of sequence isomers are presented. These studies highlight the potential of FTICR-MS as a superior technique for the analysis of combinatorially generated compounds.

Structural elucidation studies of erythromycins by electrospray tandem mass spectrometry II.

Erythromycin A (EryA), sec-butyl erythromycin B (SEryB), oleandomycin (Olean) and a synthetic derivative, roxithromycin (Rox), were used to investigate the fragmentation of polyketide macrolide antibiotics by collision induced dissociation (CID) tandem mass spectrometry (MS/MS). Analyses were performed with two commercially available mass spectrometers: a Q-TOF hybrid quadrupole time-of-flight instrument and a BioApex II (4.7 Tesla) Fourier transform ion cyclotron resonance (FTICR) instrument both equipped with electrospray ionisation (ESI) sources. One of the first fragmentation processes is the loss of an H(2)O molecule from the [M+H](+) ion. EryA has three hydroxyl groups on the polyketide ring and loses three H(2)O molecules during CID. This study indicates that these facts are not necessarily related. Deuterium exchange experiments were carried out in order to isotopically label free hydroxyl groups. (18)O-exchange experiments were also carried out in order to label the carbonyl group at the 9-position. In EryA and its analogue the first H(2)O loss shifts in mass from loss of 18 Da to loss of 20 Da in deuterated solvents. For both molecules the loss also shifts in mass from loss of 18 Da to loss of 20 Da during the (18)O-exchange experiments. This suggests that the first loss of H(2)O is from the 9-position carbonyl group, indicating that this, and not the nitrogen of the amino sugar, is the site of protonation of the activated MH(+) ions. For Rox the initial loss of H(2)O is replaced by loss of the 9-position oxime group, the rest of the fragmentation sequence being the same as for EryA. For Olean, there is no H(2)O loss from the parent ion. The results have allowed the proposal of a mechanism for the first loss of H(2)O in the EryA MS/MS fragmentation.

Structural elucidation studies of erythromycins by electrospray tandem mass spectrometry.

Erythromycin A (EryA) was studied by electrospray ionisation tandem mass spectrometry (ESI-MS/MS) with the aim of developing a methodology for the structural elucidation of novel erythromycins developed by biological synthetic methods. Skimmer dissociation along with sequential mass spectrometry studies (up to MS5) have been employed in this study. In the low-resolution MS/MS analysis of the polyketides, there are several fragment ions that are easily assigned to various neutral losses. These have all been confirmed by accurate-mass measurements. There is also a series of peaks due to ring opening and fragmentation that can only be assigned by high-resolution MSn analysis. Further experiments were performed in deuterated media (D2O/CD3OD 50%) which, along with the high-resolution MSn of erythromycin analogues, has enabled us to identify some of the steps in the ring fragmentation, particularly the loss of the polyketide starter acid. This is an essential step for determining structural alterations in the novel polyketides, but further labelling experiments and studies on more erythromycin analogues are required before the complete fragmentation pathway can be confirmed.

Effects of melatonin, progestagens, and the ram on out-of-season reproduction in Swedish Landrace finewool sheep.

One hundred twenty-one Swedish Landrace Finewool ewes were treated with progestagen sponges (P), teaser ram stimulation (R), or melatonin implants plus teaser ram stimulation (M) in preparation for breeding with whole rams in August. Blood progesterone analyses from ewes in the R and M groups gave no evidence of luteal activity before the introduction of teaser rams. There were no significant differences between treatments for pregnancy rate (approximately 90%). The P group had the most compact lambing season, while median breeding dates for M and R groups were delayed by one cycle. In those groups, the introduction of breeding rams was later found to have been too late. M and R differed significantly for probable conception date but not for lambing dates. Circa 30% of M ewes did not have a short 6 day ovulation cycle after the first ovulation, which resulted in a less concentrated lambing season than the other methods. Although no significant differences in litter size were seen among the 3 treatments, M had the highest group average, 2.25. The ewes in this study were not in very deep anestrous in the middle of August. This supports the conclusion that treatment with exogenous hormones is not necessary to breed Swedish Landrace Finewool ewes successfully in late August/early September.

A cDNA encoding an S-locus specific glycoprotein from Brassica oleracea plants containing the S5 self-incompatibility allele.

A cDNA sequence homologous to the Brassica self-incompatibility locus specific glycoprotein (SLSG) sequence was isolated from stigmas of B. oleracea plants homozygous for the S5 allele. The nucleotide sequence of this cDNA was obtained and compared with the S6 allelic form of the SLSG. Evidence is presented which indicates that this sequence does not specify the self-incompatibility response of pollen.