Generation of new landomycins with altered saccharide patterns through over-expression of the glycosyltransferase gene lanGT3 in the biosynthetic gene cluster of landomycin A in Streptomyces cyanogenus S-136

Two novel landomycin compounds, landomycins I and J, were generated with a new mutant strain of Streptomyces cyanogenus in which the glycosyltransferase that is encoded by lanGT3 was over-expressed. This mutant also produced the known landomycins A, B, and D. All these compounds consist of the same polyketide-derived aglycon but differ in their sugar moieties, which are chains of different lengths. The major new metabolite, landomycin J, was found to consist of landomycinone with a tetrasaccharide chain attached. Combined with previous results of the production of landomycin E (which contains three sugars) by the LanGT3- mutant strain (obtained by targeted gene deletion of lanGT3), it was verified that LanGT3 is a D-olivosyltransferase responsible for the transfer of the fourth sugar required for landomycin A biosynthesis. The experiments also showed that gene over-expression is a powerful method for unbalancing biosynthetic pathways in order to generate new metabolites. The cytotoxicity of the new landomycins--compared to known ones--was assessed by using three different tumor cell lines, and their structure-activity relationship (SAR) with respect to the length of the deoxysugar side chain was deduced from the results.     

Defensive Secretions of the Carabid Beetle <Emphasis Type="Italic">Chlaenius cordicollis</Emphasis>: Chemical Components and their Geographic Patterns of Variation

The defensive secretion of the ground beetle Chlaenius cordicollis is predominantly 3-methylphenol. Adult C. cordicollis were collected in Pennsylvania and Manitoba and induced to discharge defensive secretion in a vial. The headspace was sampled by solid phase microextraction, and samples were analyzed by gas chromatography–mass spectrometry. Five alkylphenolic compounds were detected: all beetles secreted 3-methlyphenol, 2,5-dimethylphenol, and 3-ethylphenol, and most beetles from each locality secreted detectable amounts of 2,3-dimethlyphenol and 3,4-dimethylphenol. In about 80% of beetles, we detected small amounts of the alkoxyphenolic compounds 2-methoxy-4-methylphenol and 2-methoxy-5-methylphenol. Multivariate compositional analysis of relative peak areas of alkylphenolic compounds revealed geographic variation and sexual dimorphism in defensive secretions. Compared with samples from Manitoba, relative peak areas of samples from Pennsylvania were lower for 2,3-dimethylphenol and higher for 3-methylphenol. Sexual dimorphism was detected only in Manitoba where, compared with samples from males, relative peak areas for samples from females were higher for 2,5-dimethylphenol and lower for 3-ethylphenol. This is the first report of geographic variation in defensive secretions of carabid beetles, and it demonstrates the need for knowledge of patterns of variation before characterizing the defensive secretions of a species as a whole.     

0.23 mu m gate length MODFETs on InAlAs/InGaAs/InP heterostructure grown by MOVPE

Modulation-doped field effect transistors (MODFETs) with 0.23 mu m gate lengths have been fabricated on an InAlAs/InGaAs/InP heterostructure grown by metal organic vapour phase epitaxy (MOVPE/MOCVD). Extrinsic DC transconductance as high as 800 mS/mm, and unity current gain cutoff frequency f/sub t/ of over 120 GHz at room temperature have been achieved. These g/sub m/ and f/sub t/ values compare favourably with the best devices of similar gate length grown by molecular-beam epitaxy (MBE) and are the highest values reported for any device grown by MOVPE.<>     

A convenient method for the preparation of hapten phosphoramidites

Total midsagittal area and seven subdivisions of the corpus callosum were measured on magnetic resonance images of 114 healthy boys and girls, aged 4 to 18. Striking variability of size was noted for all measures. Total midsagittal corpus callosum area increased in a robust and linear fashion from ages 4 to 18 (slope = 13.1 mm2/year, P = 0.0001 and slope = 11.1 mm2/year, P = 0.0001 for females and males, respectively). Posterior and mid regions demonstrated greater age-related changes than anterior regions with the rostrum and genu (anterior regions) having reached adult sizes in the youngest of our subjects. There were no significant effects of sex for any measures. These findings support anatomical studies indicating ongoing myelination of higher association areas throughout adolescence, but raise intriguing questions about anterior-posterior gradients of interhemispheric myelination.     

Conditioning and experiential factors affecting the development of sensitization to apomorphine.

In 3 experiments, the role of conditioning and experiential factors in producing behavioral sensitization to apomorphine (APO) was examined. In each experiment, male rats received intermittent injections of APO (5.0 mg/kg s.c.) or vehicle and were tested for locomotor activity in photocell arenas. Activity test experience was paired or unpaired with drug exposure or not given. After the pretreatment phase in each experiment, all rats were tested for activity after an APO injection. The results indicated that behavioral sensitization to APO develops with repeated treatments in the absence of drug-associated contextual environmental stimuli. The magnitude of the sensitization effect observed, however, was always greater in rats for which specific environmental cues were reliably associated with drug exposure. These findings indicate that behavioral sensitization to APO develops through both associational and non-associational mechanisms. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Immunoassay reagents for thyroid testing. 2. Binding properties and energetic parameters of a T4 monoclonal antibody and its Fab fragment with a library of thyroxine analog biosensors using surface plasmon resonance

Over the past decade only a limited number of public health initiatives have been subjected to systematic monitoring and evaluation and, in many instances, there is growing pressure to estimate which approaches work best for a given level of inputs in order to allocate resources effectively. However, evaluation is very often seen as punitive, and a change in perception is needed to allow evaluation to be owned by all stakeholders in public programs. In the field of HIV/AIDS prevention and care, the first difficulty is that many national AIDS programs lack clearly stated objectives and involve a wide variety of players. These players each have their own guidelines for project/program design, monitoring, and evaluation. The second difficulty relates to the fact that evaluation involves "multiple methods, multiple audiences, multiple funding sources, multiple perspectives, multiple paradigms, multiple roles, and multiple solutions to multiple problems" (Quinn Patton, 1986). To some people, evaluation calls for complex experimental studies while to others it means pausing at the end of an activity to sort out what went well and what went less successfully. This paper examines briefly some of the problems and challenges facing the evaluation of HIV prevention and care and summarizes the approaches adopted by the World Health Organization (WHO) to assist AIDS programs around the world in evaluating their initiatives. The paper also provides an update on the progress of developments, training, and implementation of these approaches.     

The impact of host immune status on the within-host and population dynamics of antigenic immune escape

Antigenically evolving pathogens such as influenza viruses are difficult to control owing to their ability to evade host immunity by producing immune escape variants. Experimental studies have repeatedly demonstrated that viral immune escape variants emerge more often from immunized hosts than from naive hosts. This empirical relationship between host immune status and within-host immune escape is not fully understood theoretically, nor has its impact on antigenic evolution at the population level been evaluated. Here, we show that this relationship can be understood as a trade-off between the probability that a new antigenic variant is produced and the level of viraemia it reaches within a host. Scaling up this intra-host level trade-off to a simple population level model, we obtain a distribution for variant persistence times that is consistent with influenza A/H3N2 antigenic variant data. At the within-host level, our results show that target cell limitation, or a functional equivalent, provides a parsimonious explanation for how host immune status drives the generation of immune escape mutants. At the population level, our analysis also offers an alternative explanation for the observed tempo of antigenic evolution, namely that the production rate of immune escape variants is driven by the accumulation of herd immunity. Overall, our results suggest that disease control strategies should be further assessed by considering the impact that increased immunity—through vaccination—has on the production of new antigenic variants.     

Providing the missing link: the exposure science ontology ExO

Environmental health information resources lack exposure data required to translate molecular insights, elucidate environmental contributions to diseases, and assess human health and ecological risks. We report development of an Exposure Ontology, ExO, designed to address this information gap by facilitating centralization and integration of exposure data. Major concepts were defined and the ontology drafted and evaluated by a working group of exposure scientists and other ontology and database experts. The resulting major concepts forming the basis for the ontology are "exposure stressor", "exposure receptor", "exposure event", and "exposure outcome". Although design of the first version of ExO focused on human exposure to chemicals, we anticipate expansion by the scientific community to address exposures of human and ecological receptors to the full suite of environmental stressors. Like other widely used ontologies, ExO is intended to link exposure science and diverse environmental health disciplines including toxicology, epidemiology, disease surveillance, and epigenetics.     

Inactivation of the Ketoreductase gilU Gene of the Gilvocarcin Biosynthetic Gene Cluster Yields New Analogues with Partly Improved Biological Activity

Four new analogues of the gilvocarcin‐type aryl‐C‐glycoside antitumor compounds, namely 4′‐hydroxy gilvocarcin V (4′‐OH‐GV), 4′‐hydroxy gilvocarcin M, 4′‐hydroxy gilvocarcin E and 12‐demethyl‐defucogilvocarcin V, were produced through inactivation of the gilU gene. The 4′‐OH‐analogues showed improved activity against lung cancer cell lines as compared to their parent compounds without 4′‐OH group (gilvocarcins V and E). The structures of the sugar‐containing new mutant products indicate that the enzyme GilU acts as an unusual ketoreductase involved in the biosynthesis of the C‐glycosidically linked deoxysugar moiety of the gilvocarcins. The structures of the new gilvocarcins indicate substrate flexibility of the post‐polyketide synthase modifying enzymes, particularly the C‐glycosyltransferase and the enzyme responsible for the sugar ring contraction. The results also shed light into biosynthetic sequence of events in the late steps of biosynthetic pathway of gilvocarcin V.     

Optimal actuator placement for large scale systems: a reduced-order modelling approach

Because of advances in surgical and cardiopulmonary bypass techniques it is now possible to definitively repair the vast majority of congenital heart disease in infancy or childhood. Although the majority of survivors do not have obvious cerebral sequelae, there is increasing disquiet about the high incidence of acute neurological events in the immediated postoperative period as well as evidence that at long-term follow-up there are subtle cognitive and motor deficits in many. Some children are more at risk of neurodevelopmental problems, either because of their cardiac (e.g. , extensive aortopulmonary collaterals) or cerebrovascular (e.g., the propensity to large vessel dissection) anatomy or because of genetic predisposition (e.g., to prothrombotic disorders). The incidence may vary with the surgery (e.g., the Fontan operation) and the cardiopulmonary bypass technique necessary to achieve an adequate technical repair (e.g., low or no flow at deep hypothermia). Recognition of the population at risk will lead to prevention of serious sequelae. Data collected in adults may be misleading, and many pediatric units have developed their own practice, but recent studies in animal models of child surgery and in children have produced some evidence to guide management to ensure the optimal cerebral as well as cardiac outcome. Pump flow should be maintained at least 30 ml/kg/min where possible, with inotropic support to maintain blood pressure if necessary. If pump flow must be lowered or circulatory arrest is essential, thorough cerebral cooling to deep hypothermic temperatures is mandatory; a pH-stat strategy may make this easier, but an alpha-stat strategy may be better in those operations that can be performed at moderate hypothermia. There is no evidence that the available pulsatile pumps offer an advantage. Tissue oxygenation may reach critical levels and a high hematocrit and oxygen tension may reduce the risk of significant hypoxia. There is a risk of embolization in children, which can be reduced with membrane oxygenators and careful monitoring; the role of arterial filtration remains controversial. The only protective agent that can currently be recommended is methylprednisolone to protect the spinal cord (e.g., in operations on the aortic arch). Further studies are needed in this important area.