IRAK-4 inhibitors. Part III: a series of imidazo[1,2-a]pyridines

Following the identification of a potent IRAK inhibitor through routine project cross screening, a novel class of IRAK-4 inhibitor was established. The SAR of imidazo[1,2-a]pyridino-pyridines and benzimidazolo-pyridines was explored.     

fMRI adaptation reveals mirror neurons in human inferior parietal cortex

Mirror neurons, as originally described in the macaque, have two defining properties [1, 2]: They respond specifically to a particular action (e.g., bringing an object to the mouth), and they produce their action-specific responses independent of whether the monkey executes the action or passively observes a conspecific performing the same action. In humans, action observation and action execution engage a network of frontal, parietal, and temporal areas. However, it is unclear whether these responses reflect the activity of a single population that represents both observed and executed actions in a common neural code or the activity of distinct but overlapping populations of exclusively perceptual and motor neurons [3]. Here, we used fMRI adaptation to show that the right inferior parietal lobe (IPL) responds independently to specific actions regardless of whether they are observed or executed. Specifically, responses in the right IPL were attenuated when participants observed a recently executed action relative to one that had not previously been performed. This adaptation across action and perception demonstrates that the right IPL responds selectively to the motoric and perceptual representations of actions and is the first evidence for a neural response in humans that shows both defining properties of mirror neurons.     

Imidazole piperazines: SAR and development of a potent class of cyclin-dependent kinase inhibitors with a novel binding mode

A piperazine series of cyclin-dependent kinase (CDK) inhibitors have been identified. The compounds exhibit excellent physiochemical properties and a novel binding mode, whereby a bridging interaction via a water molecule with Asp 86 of CDK2, leads to selectivity for the CDK family of enzymes over other kinases. Piperazines 2e and 2i were subsequently shown to inhibit tumour growth when dosed orally in a nude mouse xenograft study. Additional chemical series that exploit this unexpected interaction with Asp 86 are also described.     

The biochemistry and biology of extracellular plant lipid-transfer proteins (LTPs)

Plant lipid-transfer proteins (LTPs) are abundant, small, lipid binding proteins that are capable of exchanging lipids between membranes in vitro. Despite their name, a role in intracellular lipid transport is considered unlikely, based on their extracellular localization. A number of other biological roles, including antimicrobial defense, signaling, and cell wall loosening, have been proposed, but conclusive evidence is generally lacking, and these functions are not well correlated with in vitro activity or structure. A survey of sequenced plant genomes suggests that the two biochemically characterized families of LTPs are phylogenetically restricted to seed plants and are present as substantial gene families. This review aims to summarize the current understanding of LTP biochemistry, as well as the evidence supporting the proposed in vivo roles of these proteins within the emerging post-genomic framework.     

Changes in the de novo, salvage, and degradation pathways of pyrimidine nucleotides during tobacco shoot organogenesis.

Pyrimidine nucleotide metabolism was studied in tobacco callus cultured for 21days under shoot-forming (SF) and non-shoot-forming (NSF) conditions by following the metabolic fate of orotic acid, a precursor of the de novo pathway, and uridine and uracil, intermediates of the salvage and degradation pathways respectively. Nucleic acid synthesis was also investigated by measuring the incorporation of labeled thymidine into different cellular components. Our results indicate that with respect to nucleotide metabolism, the organogenic process in tobacco can be divided in two "metabolic phases": a de novo phase followed by a salvage phase. The initial stages of meristemoid formation during tobacco organogenesis (up to day 8) are characterized by a heavy utilization of orotic acid into nucleotides and nucleic acids. Utilization of this intermediate for the de novo synthesis of nucleotides, which is limited in NSF tissue, is mainly due to the activity of orotate phosphoribosyltransferase (OPRT), which increases in tissue cultured under SF conditions. After day 8, nucleotide synthesis during shoot growth seems to be mainly due to the salvage activity of both uridine and uracil. Both intermediates are preferentially utilized in SF tissue for the formation of nucleotides and nucleic acids through the activities of their respective salvage enzymes: uridine kinase (URK), and uracil phosphoribosyltransferase (UPRT). Metabolic studies on thymidine indicate that in SF tissue maximal nucleic acid synthesis occurs at day 4, in support of the initiation of meristemoid formation. Overall these results suggest that the organogenic process in tobacco is underlined by precise fluctuations in pyrimidine metabolism which delineate structural events culminating in shoot formation.     

Lethal and sublethal effects of spinosad-based GF-120 bait on the tephritid parasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae)

Experiments were performed to determine the likely compatibility of spinosad-based GF-120 applications and mass-releases of the braconid parasitoid Diachasmimorpha longicaudata (Ashmead) for control of tephritid fruit flies. Severe effects on D. longicaudata survival and reproduction in the laboratory contrasted with mild effects in semi-field studies. Topical application and contact with dried residues on plastic surfaces in the laboratory generally resulted in high mortality of wasps and significantly reduced survival times compared to controls. Survival times of both sexes were more severely affected when exposure to dried residues on mango leaves compared to wasps that were offered GF-120 mixed with honey. Brief (24 h) exposure to GF-120 in honey or dried residues on mango leaves did not affect total progeny production by wasps, whereas continuous exposure over a 10 d period resulted in reductions in female survival, progeny production and net fecundity (∑L_xM_x) compared to honey-fed controls. During the rainy season, wasp survival times in field cages with young mango trees that had been sprayed with GF-120 or treated with GF-120 mixed with honey were only reduced by 1 or 2 days, respectively, compared to average 10.1 d survival of honey-fed controls. During the dry season, wasp mortality was not significantly affected by exposure to GF-120 residues on mango leaves that had been subjected to natural weathering in the field, compared to controls. Our results suggest that applications of GF-120 are unlikely to adversely influence the effectiveness of mass-releases of D. longicaudata. This prediction requires validation from comparative field studies on parasitoid populations in treated and untreated areas.     

Development and mapping of EST-derived simple sequence repeat markers for hexaploid wheat.

Expressed sequence tags (ESTs) are a valuable source of molecular markers. To enhance the resolution of an existing linkage map and to identify putative functional polymorphic gene loci in hexaploid wheat (Triticum aestivum L.), over 260,000 ESTs from 5 different grass species were analyzed and 5418 SSR-containing sequences were identified. Using sequence similarity analysis, 156 cross-species superclusters and 138 singletons were used to develop primer pairs, which were then tested on the genomic DNA of barley (Hordeum vulgare), maize (Zea mays), rice (Oryza sativa), and wheat. Three-hundred sixty-eight primer pairs produced PCR amplicons from at least one species and 227 primer pairs amplified DNA from two or more species. EST-SSR sequences containing dinucleotide motifs were significantly more polymorphic (74%) than those containing trinucleotides (56%), and polymorphism was similar for markers in both coding and 5' untranslated (UTR) regions. Out of 112 EST-SSR markers, 90 identified 149 loci that were integrated into a reference wheat genetic map. These loci were distributed on 19 of the 21 wheat chromosomes and were clustered in the distal chromosomal regions. Multiple-loci were detected by 39% of the primer pairs. Of the 90 mapped ESTs, putative functions for 22 were identified using BLASTX queries. In addition, 80 EST-SSR markers (104 loci) were located to chromosomes using nullisomic-tetrasomic lines. The enhanced map from this study provides a basis for comparative mapping using orthologous and PCR-based markers and for identification of expressed genes possibly affecting important traits in wheat.     

EVIDENCE FOR INCREASES IN ANTARCTIC BLUE WHALES BASED ON BAYESIAN MODELLING

Antarctic blue whales ( Balaenoptera musculus intermedia ) are the largest and formerly most abundant blue whale subspecies, but were hunted to near extinction last century. Estimated whaling mortality was unsustainable from 1928 to 1972 (except during 1942–1944), depleting them from 239,000 (95% interval 202,000–311,000) to a low of 360 (150–840) in 1973. Obtaining statistical evidence for subsequent increases has proved difficult due to their scarcity. We fitted Bayesian models to three sighting series (1968–2001), constraining maximum rates of increase to 12% per annum. These models indicated that Antarctic blue whales are increasing at a mean rate of 7.3% per annum (1.4%–11.6%). Informative priors based on blue whale biology (4.3%, SD = 1.9%) and a Bayesian hierarchical meta-analysis of increase rates in other blue whale populations (−3%, SD = 11.6%), suggest plausible increase rates are lower (although the latter has wide intervals), but a meta-analysis of other mysticetes obtains similar rates of increase (6.7%, SD = 4.0%). Possible biases affecting the input abundance estimates are discussed. Although Antarctic blue whales appear to have been increasing since Sovier illegal whaling ended in 1972, they still need to be protected-their estimated 1996 population size, 1,700 (860–2,900), was just 0.7% (0.3%–1.3%) of the pre-exploitation level.     

Temporal and Spatial Occurrence of Female Chinook Salmon Carrying a Male-Specific Genetic Marker in the Columbia River Watershed

Recent declines in many chinook salmon, Oncorhynchus tshawytscha, populations within the Columbia River watershed have prompted an examination of their reproductive biology. In a previous study many female fall chinook salmon collected in 1999 from the Hanford Reach of the Columbia River tested positive for a male-specific DNA marker (OtY1) found on the Y chromosome. The purpose of this study was to determine if females testing positive for the OtY1 marker could be found in other populations of fall chinook salmon from the Columbia River, and to assess the prevalence of OtY1 incidence in different female cohorts. Post-spawned male and female fall chinook salmon from three different naturally spawning populations (Hanford Reach, Yakima River and Ives Island) and one hatchery population (Priest Rapids Hatchery) on the Columbia River were tested in 2000 and 2001 for the OtY1 marker. Among naturally spawning populations, 57.4% of the females tested positive from the Hanford Reach, 33.3% tested positive from the Yakima River, and 32.5% tested positive from Ives Island. Of the Priest Rapids Hatchery fish, 62.5% of the females tested positive, and significant differences were detected between the 1995–1996 and 1997–1998 female cohorts from this population. No significant differences were detected between any of the female cohorts from the naturally spawning populations. All male chinook salmon samples, tested positive for OtY1.     

Cortico-striatal representation of time in animals and humans

Interval timing in the seconds-to-minutes range is crucial to learning, memory, and decision-making. Recent findings argue for the involvement of cortico-striatal circuits that are optimized by the dopaminergic modulation of oscillatory activity and lateral connectivity at the level of cortico-striatal inputs. Striatal medium spiny neurons are proposed to detect the coincident activity of specific beat patterns of cortical oscillations, thereby permitting the discrimination of supra-second durations based upon the reoccurring patterns of subsecond neural firing. This proposal for the cortico-striatal representation of time is consistent with the observed psychophysical properties of interval timing (e.g. linear time scale and scalar variance) as well as much of the available pharmacological, lesion, patient, electrophysiological, and neuroimaging data from animals and humans (e.g. dopamine-related timing deficits in Huntington's and Parkinson's disease as well as related animal models). The conclusion is that although the striatum serves as a 'core timer', it is part of a distributed timing system involving the coordination of large-scale oscillatory networks.