Expression cloning and periplasmic orientation of the Francisella novicida lipid A 4'-phosphatase LpxF

Francisella tularensis and related intracellular pathogens synthesize lipid A molecules that differ from their Escherichia coli counterparts. Although a functional orthologue of lpxK, the gene encoding the lipid A 4'-kinase, is present in Francisella, no 4'-phosphate moiety is attached to Francisella lipid A. We now demonstrate that a membrane-bound phosphatase present in Francisella novicida U112 selectively removes the 4'-phosphate residue from tetra- and pentaacylated lipid A molecules. A clone that expresses the F. novicida 4'-phosphatase was identified by assaying lysates of E. coli colonies, harboring members of an F. novicida genomic DNA library, for 4'-phosphatase activity. Sequencing of a 2.5-kb F. novicida DNA insert from an active clone located the structural gene for the 4'-phosphatase, designated lpxF. It encodes a protein of 222 amino acid residues with six predicted membrane-spanning segments. Rhizobium leguminosarum and Rhizobium etli contain functional lpxF orthologues, consistent with their lipid A structures. When F. novicida LpxF is expressed in an E. coli LpxM mutant, a strain that synthesizes pentaacylated lipid A, over 90% of the lipid A molecules are dephosphorylated at the 4'-position. Expression of LpxF in wild-type E. coli has no effect, because wild-type hexaacylated lipid A is not a substrate. However, newly synthesized lipid A is not dephosphorylated in LpxM mutants by LpxF when the MsbA flippase is inactivated, indicating that LpxF faces the outer surface of the inner membrane. The availability of the lpxF gene will facilitate re-engineering lipid A structures in diverse bacteria.     

Evidence that complement protein C1q interacts with C-reactive protein through its globular head region

C1q acts as the recognition unit of the first complement component, C1, and binds to immunoglobulins IgG and IgM, as well as to non-Ig ligands, such as C-reactive protein (CRP). IgG and IgM are recognized via the globular head regions of C1q (C1qGR), whereas CRP has been postulated to interact with the collagen-like region (C1qCLR). In the present study, we used a series of nine mAbs to C1q, five directed against C1qGR and four against C1qCLR, to inhibit the interaction of C1q with CRP. The F(ab')(2) of each of the five mAbs directed against C1qGR inhibited binding of C1q to polymerized IgG. These five mAbs also successfully inhibited the interaction of C1q with CRP. Moreover, these five mAbs inhibited C1 activation by CRP as well as by polymerized IgG in vitro. In contrast, none of the four mAbs against C1qCLR inhibited C1q interaction with CRP or IgG, or could reduce activation of complement by CRP or polymerized IgG. These results provide the first evidence that the interaction of C1q with CRP or IgG involves sites located in the C1qGR, whereas sites in the CLR do not seem to be involved in the physiological interaction of C1q with CRP.     

Structure-guided design of peptide-based tryptase inhibitors

Improved peptide-based inhibitors of human beta tryptase were discovered using information gleaned from tripeptide library screening and structure-guided design methods, including fragment screening. Our efforts sought to improve this class of inhibitors by replacing the traditional Lys or Arg P1 element. The optimized compounds display low nanomolar potency against the mast cell target and several hundred-fold selectivity with respect to serine protease off targets. Thus, replacement of Lys/Arg at P1 in a peptide-like scaffold does not need to be accompanied by a loss in target affinity.     

Resilience of inshore, juvenile snapper Pagrus auratus to angling and release

This study assessed the mortality of 157 snapper Pagrus auratus (9-29 cm, total length, L(T) ) after being conventionally angled and then released into cages (along with 48 controls) for 4 days off south-eastern Australia. Fatalities were restricted to 12 angled fish (7·6%) and mostly attributed to the ingestion of hooks and especially their subsequent removal, which caused substantial blood loss and immediate death. Hook ingestion was significantly biased towards smaller fish (<21 cm L(T)) and attributed to a lower chance of anglers initially detecting these individuals on the line (allowing them to consume more of the baits). While mortalities might be reduced in future via (1) choosing terminal rigs that promote mouth hooking and (2) cutting the line on any-hook ingested fish, the results nevertheless validate releasing unwanted angled inshore juvenile P. auratus as a means for managing their exploitation.     

Long-term removal of wheat straw decreases soil amorphous silica at Broadbalk, Rothamsted

Aims

Most cereals accumulate Si in their shoots. Soil bioavailability of Si may be a constraint on the beneficial role of silica in cereals but it is not yet well supported by field data. The aim of this study is to evaluate the long-term impact of wheat straw exports on the pool of soil phytoliths, which, it is suggested, represents the most labile and renewable pool of soil Si.

Methods

We measured the amorphous Si (ASi) in soils from several experiments at Rothamsted Research (UK), which provided long-term soil data back to the middle of the 19th century, using two alternative extraction techniques: Na₂CO₃ (referred to as AS_nc) or zinc bromide extraction (referred to as ASi_zb).

Results

All samples showed a similar range of AS_nc and ASi_zb but low values (0.1–3.4?mg?g^?1 DW) compared to published data on natural ecosystems. In the Broadbalk experiment, a decrease over time in ASi in the topsoil samples is in good agreement with the hypothesis that cropping and exports of straw leads to depletion of soil phytoliths. A decrease in Si concentration in straw samples was observed between 1883 and 1944. From 1944 to the present, Si concentration increased irregularly in the straw, probably as the result of liming, which enhanced the dissolution of the remaining phytoliths through increasing pH. In the reforested Geescroft field the higher phytolith concentration in the modern topsoil samples is in good agreement with a re-building of phytolith storage from litter input in an acidic environment.

Conclusions

Our results therefore support the hypothesis that export of wheat straw leads to a decrease in bioavailable Si.     

A study of trait anhedonia in non-clinical Chinese samples: evidence from the Chapman Scales for Physical and Social Anhedonia

Background

Recent studies suggest that anhedonia, an inability to experience pleasure, can be measured as an enduring trait in non-clinical samples. In order to examine trait anhedonia in a non-clinical sample, we examined the properties of a range of widely used questionnaires capturing anhedonia.

Methods

887 young adults were recruited from colleges. All of them were administered a set of checklists, including Chapman Scale for Social Anhedonia (CRSAS) and the Chapman Scale for Physical Anhedonia Scale (CPAS), The Temporal Experience of Pleasure Scale(TEPS), and The Schizotypal Personality Questionnaire (SPQ).

Results

Males showed significantly higher level of physical (F = 5.09, p<0.001) and social (F = 4.38, p<0.005) anhedonia than females. As expected, individuals with schizotypal personality features also demonstrated significantly higher scores of physical (t = 3.81, p<0.001) and social (t = 7.33, p<0.001) trait anhedonia than individuals without SPD features, but no difference on self-report anticipatory and consummatory pleasure experience.

Conclusions

Concerning the comparison on each item of physical and social anhedonia, the results indicated that individuals with SPD feature exhibited higher than individuals without SPD features on more items of social anhedonia than physical anhedonia scale. These preliminary findings suggested that trait anhedonia can be identified a non-clinical sample. Exploring the demographic and clinical correlates of trait anhedonia in the general population may provide clues to the pathogenesis of psychotic disorder.     

Are Neoclassical Canons Valid for Southern Chinese Faces?

Background

Proportions derived from neoclassical canons, initially described by Renaissance sculptors and painters, are still being employed as aesthetic guidelines during the clinical assessment of the facial morphology.

Objective

1. to determine the applicability of neoclassical canons for Southern Chinese faces and 2. to explore gender differences in relation to the applicability of the neoclassical canons and their variants.

Methodology

3-D photographs acquired from 103 young adults (51 males and 52 females) without facial dysmorphology were used to test applicability of four neoclassical canons. Standard anthropometric measurements that determine the facial canons were made on these 3-D images. The validity of the canons as well as their different variants were quantified.

Principal Findings

The neoclassical cannons seldom applied to these individuals, and facial three-section and orbital canons did not apply at all. The orbitonasal canon was most frequently applicable, with a frequency of 19%. Significant sexual dimorphism was found relative to the prevalence of the variants of facial three-section and orbitonasal canons.

Conclusion

The neoclassical canons did not appear to apply to our sample when rigorous quantitative measurements were employed. Thus, they should not be used as esthetic goals for craniofacial surgical interventions.     

Implication of new WHO growth standards on identification of risk factors and estimated prevalence of malnutrition in rural Malawian infants

Background

The World Health Organization (WHO) released new Child Growth Standards in 2006 to replace the current National Center for Health Statistics (NCHS) growth reference. We assessed how switching from the NCHS to the newly released WHO Growth Standards affects the estimated prevalence of wasting, underweight and stunting, and the pattern of risk factors identified.

Methodology/Principal Findings

Data were drawn from a village-informant driven Demographic Surveillance System in Northern Malawi. Children (n = 1328) were visited twice at 0–4 months and 11–15 months. Data were collected on the demographic and socio-economic environment of the child, health history, maternal and child anthropometry and child feeding practices. Weight-for-length, weight-for-age and length-for-age were derived in z-scores using the two growth references. In early infancy, prevalence estimates were 2.9, 6.1, and 8.5 fold higher for stunting, underweight, and wasting respectively using the WHO standards compared to NCHS reference (p<0.001 for all). At one year, prevalence estimates for wasting and stunting did not differ significantly according to reference used, but the prevalence of underweight was half that with the NCHS reference (p<0.001). Patterns of risk factors were similar with the two growth references for all outcomes at one year although the strength of association was higher with WHO standards.

Conclusions/Significance

Differences in prevalence estimates differed in magnitude but not direction from previous studies. The scale of these differences depends on the population''s nutritional status thus it should not be assumed a priori. The increase in estimated prevalence of wasting in early infancy has implications for feeding programs targeting lactating mothers and ante-natal multiple micronutrients supplementation to tackle small birth size. Risk factors identified using WHO standards remain comparable with findings based on the NCHS reference in similar settings. Further research should aim to identify whether the young infants additionally diagnosed as malnourished by this new standard are more appropriate targets for interventions than those identified with the NCHS reference.     

Pharmacokinetic genes do not influence response or tolerance to citalopram in the STAR*D sample

Background

We sought to determine whether clinical response or tolerance to the Selective Serotonin Reuptake Inhibitor (SSRI) citalopram is associated with genetic polymorphisms in potentially relevant pharmacokinetic enzymes.

Methodology

We used a two-stage case-control study design in which we split the sample of 1,953 subjects from the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) trial into a discovery (n = 831) and validation set (n = 1,046). Fifteen polymorphisms from five (CYP2D6, ABCB1, CYP2C19, CYP3A4, and CYP3A5) pharmacokinetic genes were genotyped. We examined the associations between these polymorphisms and citalopram response and tolerance. Significant associations were validated in the second stage for those polymorphism found to be statistically significant in the first stage.

Conclusions

No genetic polymorphism in the pharmacokinetic genes examined was significantly associated with our response or tolerance phenotypes in both stages. For managing pharmacological treatment with citalopram, routine screening of the common pharmacokinetic DNA variants that we examined appears to be of limited clinical utility.