Accumulation of Phosphatidic Acid Increases Vancomycin Resistance in Escherichia coli

In Gram-negative bacteria, lipopolysaccharide (LPS) contributes to the robust permeability barrier of the outer membrane, preventing entry of toxic molecules such as antibiotics. Mutations in lptD, the beta-barrel component of the LPS transport and assembly machinery, compromise LPS assembly and result in increased antibiotic sensitivity. Here, we report rare vancomycin-resistant suppressors that improve barrier function of a subset of lptD mutations. We find that all seven suppressors analyzed mapped to the essential gene cdsA, which is responsible for the conversion of phosphatidic acid to CDP-diacylglycerol in phospholipid biosynthesis. These cdsA mutations cause a partial loss of function and, as expected, accumulate phosphatidic acid. We show that this suppression is not confined to mutations that cause defects in outer membrane biogenesis but rather that these cdsA mutations confer a general increase in vancomycin resistance, even in a wild-type cell. We use genetics and quadrupole time of flight (Q-TOF) liquid chromatography-mass spectrometry (LC-MS) to show that accumulation of phosphatidic acid by means other than cdsA mutations also increases resistance to vancomycin. We suggest that increased levels of phosphatidic acid change the physical properties of the outer membrane to impede entry of vancomycin into the periplasm, hindering access to its target, an intermediate required for the synthesis of the peptidoglycan cell wall.     

A Statistical Model for Regional Tornado Climate Studies

Tornado reports are locally rare, often clustered, and of variable quality making it difficult to use them directly to describe regional tornado climatology. Here a statistical model is demonstrated that overcomes some of these difficulties and produces a smoothed regional-scale climatology of tornado occurrences. The model is applied to data aggregated at the level of counties. These data include annual population, annual tornado counts and an index of terrain roughness. The model has a term to capture the smoothed frequency relative to the state average. The model is used to examine whether terrain roughness is related to tornado frequency and whether there are differences in tornado activity by County Warning Area (CWA). A key finding is that tornado reports increase by 13% for a two-fold increase in population across Kansas after accounting for improvements in rating procedures. Independent of this relationship, tornadoes have been increasing at an annual rate of 1.9%. Another finding is the pattern of correlated residuals showing more Kansas tornadoes in a corridor of counties running roughly north to south across the west central part of the state consistent with the dryline climatology. The model is significantly improved by adding terrain roughness. The effect amounts to an 18% reduction in the number of tornadoes for every ten meter increase in elevation standard deviation. The model indicates that tornadoes are 51% more likely to occur in counties served by the CWAs of DDC and GID than elsewhere in the state. Flexibility of the model is illustrated by fitting it to data from Illinois, Mississippi, South Dakota, and Ohio.     

Effects of vertebrate growth factors on digestive gland cells from the mollusc Pecten maximus L.: an in vitro study

In relation with the digestive cycle, the digestive gland cells of bivalve molluscs undergo a sequence of cytological changes which is controlled by external and internal effectors such as putative gastrointestinal hormones and growth differentiation factors. A tissue dissociation method was developed to investigate the in vitro effect of the vertebrate growth and differentiation factors: insulin, insulin growth factor I (IGF-I), basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) on the digestive gland cells of the scallop Pecten maximus. All these vertebrate peptides induced a dose-dependent increased incorporation of ³H-leucine and ¹⁴C-uridine in whole digestive gland cell suspensions. However, after Percoll density gradient purification of the digestive cells, only stem and undifferentiated enriched cell fractions were responsive to the different peptides. In addition, insulin and IGF-I, but not EGF and bFGF, stimulated ³H-leucine incorporation in control dispersed mantle edge cells. These results suggest that insulin-related peptides could work as general growth promoting factors in molluscs. On the other hand, EGF and bFGF, or at least their molluscan counterparts, may be efficient growth differentiation factors in the regenerative processes occurring in the digestive gland of molluscs. Accepted: 26 September 1997     

Associations between Domestic-Dog Morphology and Behaviour Scores in the Dog Mentality Assessment

The domestic dog shows a wide range of morphologies, that humans have selected for in the process of creating unique breeds. Recent studies have revealed correlations between changes in morphology and behaviour as reported by owners. For example, as height and weight decrease, many undesirable behaviours (non-social fear, hyperactivity and attention seeking) become more apparent. The current study aimed to explore more of these correlations, but this time used reports from trained observers. Phenotypic measurements were recorded from a range of common dog breeds (n = 45) and included cephalic index (CI: the ratio of skull width to skull length), bodyweight, height and sex. These data were then correlated with results from the Dog Mentality Assessment (DMA), which involves trained observers scoring a dog’s reaction to stimuli presented over 10 standardised subtests. Each subtest is designed to evoke a behavioural response. Backward elimination and weighted step-wise regression revealed that shorter dogs demonstrated more aggressive tendencies, reacting defensively toward both assistants dressed as ghosts (p = 0.045), and to a dummy (p = 0.008). Taller dogs were more affectionate when greeting and being handled by humans (p = 0.007, p = <0.001, respectively). Taller dogs were also more cooperative (p = <0.001), and playful (p = 0.001) with humans than shorter dogs. Heavier dogs were more inquisitive toward a dummy (p = 0.011), to the source of a metallic noise (p = 0.010) and to an assistant (p = 0.003). Heavier dogs were also more attentive to the ghosts (p = 0.013). In comparison, lighter dogs were cautious of a dummy (p = <0.001) and fearful of the sound of a gunshot (p = <0.001). Lighter dogs were also cautious of, and demonstrated prolonged fearfulness toward, the source of metallic noise (p = <0.001, p = <0.034, respectively). With a far larger sample and the advantage of third-party reporting (which overcomes potential owner bias), the current findings build on previous studies in this field, further supporting covariance between morphology and behaviour.     

Quantitative Comparison of the Efficacy of Various Compounds in Lowering Intracellular Cholesterol Levels in Niemann-Pick Type C Fibroblasts

Niemann-Pick Type C disease (NPC) is a lethal, autosomal recessive disorder caused by mutations in the NPC1 and NPC2 cholesterol transport proteins. NPC’s hallmark symptoms include an accumulation of unesterified cholesterol and other lipids in the late endosomal and lysosomal cellular compartments, causing progressive neurodegeneration and death. Although the age of onset may vary in those affected, NPC most often manifests in juveniles, and is usually fatal before adolescence. In this study, we investigated the effects of various drugs, many of which modify the epigenetic control of NPC1/NPC2 gene expression, in lowering the otherwise harmful elevated intracellular cholesterol levels in NPC cells. Our studies utilized a previously described image analysis technique, which allowed us to make quantitative comparisons of the efficacy of these drugs in lowering cholesterol levels in a common NPC1 mutant model. Of the drugs analyzed, several that have been previously studied (vorinostat, panobinostat, and β-cyclodextrin) significantly lowered the relative amount of unesterified cellular cholesterol, consistent with earlier observations. In addition, a novel potential treatment, rapamycin, likewise alleviated the NPC phenotype. We also studied combinations of effective compounds with β-cyclodextrin; the addition of β-cyclodextrin significantly enhanced the cholesterol-lowering activity of vorinostat and panobinostat, but had mixed effects with rapamycin. Collectively, these results may provide a basis for the eventual development of improved NPC therapies.     

Heads First: Visual Aftereffects Reveal Hierarchical Integration of Cues to Social Attention

Determining where another person is attending is an important skill for social interaction that relies on various visual cues, including the turning direction of the head and body. This study reports a novel high-level visual aftereffect that addresses the important question of how these sources of information are combined in gauging social attention. We show that adapting to images of heads turned 25° to the right or left produces a perceptual bias in judging the turning direction of subsequently presented bodies. In contrast, little to no change in the judgment of head orientation occurs after adapting to extremely oriented bodies. The unidirectional nature of the aftereffect suggests that cues from the human body signaling social attention are combined in a hierarchical fashion and is consistent with evidence from single-cell recording studies in nonhuman primates showing that information about head orientation can override information about body posture when both are visible.