Autoclave method for rapid preparation of bacterial PCR-template DNA

An autoclave method for preparing bacterial DNA for PCR template is presented, it eliminates the use of detergents, organic solvents, and mechanical cellular disruption approaches, thereby significantly reducing processing time and costs while increasing reproducibility. Bacteria are lysed by rapid heating and depressurization in an autoclave. The lysate, cleared by microcentrifugation, was either used directly in the PCR reaction, or concentrated by ultrafiltration. This approach was compared with seven established methods of DNA template preparation from four bacterial sources which included boiling Triton X-100 and SDS, bead beating, lysozyme/proteinase K, and CTAB lysis method components. Bacteria examined were Enterococcus and Escherichia coli, a natural marine bacterial community and an Antarctic cyanobacterial-mat. DNAs were tested for their suitability as PCR templates by repetitive element random amplified polymorphic DNA (RAPD) and denaturing gradient gel electrophoresis (DGGE) analysis. The autoclave method produced PCR amplifiable template comparable or superior to the other methods, with greater reproducibility, much shorter processing time, and at a significantly lower cost.     

Changing Exploitation of Terrestrial Vertebrates During the Past 3000 Years on Tobago, West Indies

On the southern Caribbean island of Tobago, we excavated two archaeological deposits, the preceramic (ca. 2900 years old) Milford 1 site (TOB-3) and the ceramic (ca. 1200 to 900 years old) Golden Grove site (TOB-13). The non-fish bone assemblages from these sites are very different, with an emphasis at TOB-3 on sea turtles (which seldom nest today on Tobago) and the collared peccary Tayassu tajacu, a large (17–30 kg) mammal extremely rare now on Tobago. TOB-3 also yielded bones of two somewhat smaller mammals that are extinct on Tobago, the red howler monkey Alouatta seniculus and paca Agouti paca. The non-fish species at TOB-3 comprise only four reptiles and seven mammals, whereas they are more diverse (29 species) at TOB-13 and mostly represent small- to medium-sized vertebrates (<10 kg, often <1 kg), such as a toad, lizards, snakes, birds, opossum, armadillo, and eight species of rodents, as well as sea turtle and peccary, although the latter is uncommon. We interpret these differences as possibly being due to a local or island-wide scarcity of big game (peccaries) in ceramic times, which would have promoted diversification of hunting practices. Related to this may have been a more sedentary way of life for Tobago's ceramic peoples, with increased agriculture leading to more hunting near the site and less time being devoted to longer distance big-game hunting. While prehistoric anthropogenic extinction or population reduction of vertebrates is well documented on Caribbean islands to the north, our data from Tobago show that such depletions probably occurred as well on continental, land-bridge islands with more diverse faunas. Models of post-Pleistocene faunal relaxation toward a lower value for species richness should not ignore human-caused losses, which may be impossible to distinguish from non-anthropogenic losses.     

Molecular structure of fagopyritol A1 (O-alpha-D-galactopyranosyl-(1 --> 3)-D-chiro-inositol) by NMR

The molecular structure of fagopyritol A1, a novel galactopyranosyl cyclitol from buckwheat seeds, was determined to be O-alpha-D-galactopyranosyl-(1 --> 3)-D-chiro-inositol by 1H and 13C NMR. Fagopyritol A1 is a positional isomer of fagopyritol B1 (O-alpha-D-galactopyranosyl-(1 --> 2)-D-chiro-inositol), representing a different series of fagopyritol oligomers. Trimethylsilyl derivatives of both compounds have similar mass spectra, but each may be identified by different abundance ratios of fragments with m/z 305/318 and 318/319.     

N5-phosphonoacetyl-L-ornithine (PALO): a convenient synthesis and investigation of influence on regulation of amino acid biosynthetic genes in Saccharomyces cerevisiae

A scalable four-step synthesis of the ornithine transcarbamylase inhibitor N⁵-phosphonoacetyl-l-ornithine (PALO) is achieved through boroxazolidinone protection of ornithine. Investigations in the model organism Saccharomyces cerevisiae found that, in contrast to a previous report, PALO did not influence growth rate or expression of genes involved in arginine metabolism.     

(1H-imidazo[4,5-c]pyridin-2-yl)-1,2,5-oxadiazol-3-ylamine derivatives: further optimisation as highly potent and selective MSK-1-inhibitors

The novel imidazo[4,5-c]pyridine 1,2,5-oxadiazol-3-yl template affords an excellent start point for identification of inhibitors of a number of protein kinases. Here we report on its optimisation for mitogen and stress-activated protein kinase-1 (MSK-1) inhibitory activity, and selectivity over other kinases.     

Intra‐floral resource partitioning between endemic and invasive flower visitors: consequences for pollinator effectiveness

1. Sympatric flower visitor species often partition nectar and pollen and thus affect each other's foraging pattern. Consequently, their pollination service may also be influenced by the presence of other flower visiting species. Ants are solely interested in nectar and frequent flower visitors of some plant species but usually provide no pollination service. Obligate flower visitors such as bees depend on both nectar and pollen and are often more effective pollinators. 2. In Hawaii, we studied the complex interactions between flowers of the endemic tree Metrosideros polymorpha (Myrtaceae) and both, endemic and introduced flower‐visiting insects. The former main‐pollinators of M. polymorpha were birds, which, however, became rare. We evaluated the pollinator effectiveness of endemic and invasive bees and whether it is affected by the type of resource collected and the presence of ants on flowers. 3. Ants were dominant nectar‐consumers that mostly depleted the nectar of visited inflorescences. Accordingly, the visitation frequency, duration, and consequently the pollinator effectiveness of nectar‐foraging honeybees (Apis mellifera) strongly decreased on ant‐visited flowers, whereas pollen‐collecting bees remained largely unaffected by ants. Overall, endemic bees (Hylaeus spp.) were ineffective pollinators. 4. The average net effect of ants on pollination of M. polymorpha was neutral, corresponding to a similar fruit set of ant‐visited and ant‐free inflorescences. 5. Our results suggest that invasive social hymenopterans that often have negative impacts on the Hawaiian flora and fauna may occasionally provide neutral (ants) or even beneficial net effects (honeybees), especially in the absence of native birds.     

Mimicking a semi-arid tropical environment achieves dormancy alleviation for seeds of Australian native Goodeniaceae and Asteraceae

Background and Aims: Seed physiological dormancy (PD) limits the use and conservationof some of Queensland's (Qld) native forb species. It was hypothesisedthat optimum dormancy-alleviating treatments would reflect environmentalconditions that seeds experience in situ, and this premise wastested for PD seeds of four species native to south-west Qld. Methods: High temperatures and increased rainfall during summer are characteristicof this semi-arid tropical environment. Ex situ treatments weredesigned to mimic conditions that seeds dispersed in springexperience during the summer months before germinating in coolerautumn temperatures. Seeds received between 4 and 20 weeks ofa dry after-ripening (DAR), warm stratification or dry/wet cyclingtreatment (DAR interspersed with short periods of warm stratification),in darkness, before being transferred to germination test conditions.In addition, natural dormancy alleviation of one of the Goodeniaceaespecies was investigated in situ. Key Results: Dry/wet cycling resulted in higher levels of germination ofActinobole uliginosum (Asteraceae), Goodenia cycloptera andVelleia glabrata (Goodeniaceae) when compared with constantDAR or stratification, while Goodenia fascicularis (Goodeniaceae)responded better to short durations of warm stratification.Long durations of DAR partially alleviated PD of A. uliginosum;however, stratification induced and maintained dormancy of thisspecies. Modifications to the dry/wet cycling treatment andgermination test conditions based on data collected in situenabled germination of G. cycloptera and V. glabrata to be furtherimproved. Conclusions: Treatments designed using temperature, relative humidity andrainfall data for the period between natural seed dispersaland germination can successfully alleviate PD. Differences betweenthe four species in conditions that resulted in maximum germinationindicate that, in addition to responding to broad-scale climatepatterns, species may be adapted to particular microsites and/orseasonal conditions.     

Warfare related trauma at Orendorf, a middle Mississippian site in west-central Illinois

Skeletal evidence of nonritual interpersonal trauma in the central Illinois valley is currently limited to the terminal prehistoric period in the region. Sixteen percent of the entire Norris Farms Oneota skeletal sample died violently, presumably because they intruded upon small groups of Mississippians who had not yet abandoned the region. Archaeological evidence of palisades, however, suggests that the region was embroiled in conflict before the Oneota arrived though the skeletal evidence supporting more than ritualized or geographically sporadic cases of scalping or embedded projectiles has been elusive. This study examines the frequency and nature of interpersonal trauma at Orendorf, a Middle Mississippian (AD 1150-1250) site at the northern periphery of the region. Nine percent (N = 25) of all 268 individuals documented at Orendorf suffered warfare-related trauma, including 13 cases of scalping, six instances of decapitation, five individuals with healed cranial blunt force trauma, three projectile point impacts, and eight cases of projectile injuries inferred by the burial context. All of the traumatized individuals were at or above the age of 15 years and males and females were victimized equally. The trauma rate among adults is 16%, which is less than that of the Norris Farms Oneota (34%) but higher than other Mississippian groups in the Southeast. The nature of the injuries is more consistent with attacks by outsiders than codified or ritualized intragroup violence.