Characterization of leptospiral outer membrane lipoprotein LipL36: downregulation associated with late-log-phase growth and mammalian infection

We report the cloning of the gene encoding a 36-kDa leptospiral outer membrane lipoprotein, designated LipL36. We obtained the N-terminal amino acid sequence of a staphylococcal V8 proteolytic-digest fragment in order to design an oligonucleotide probe. A Lambda-Zap II library containing EcoRI fragments of Leptospira kirschneri DNA was screened, and a 2.3-kb DNA fragment which contained the entire structural lipL36 gene was identified. Several lines of evidence indicate that LipL36 is lipid modified in a manner similar to that of LipL41, a leptospiral outer membrane lipoprotein we described in a previous study (E. S. Shang, T. A. Summers, and D. A. Haake, Infect. Immun. 64:2322-2330, 1996). The deduced amino acid sequence of LipL36 would constitute a 364-amino-acid polypeptide with a 20-amino-acid signal peptide, followed by an L-X-Y-C lipoprotein signal peptidase cleavage site. LipL36 is solubilized by Triton X-114 extraction of L. kirschneri; phase separation results in partitioning of LipL36 exclusively into the hydrophobic, detergent phase. LipL36 is intrinsically labeled during incubation of L. kirschneri in media containing [3H]palmitate. Processing of LipL36 is inhibited by globomycin, a selective inhibitor of lipoprotein signal peptidase. After processing, LipL36 is exported to the outer membrane along with LipL41 and lipopolysaccharide. Unlike LipL41, there appears to be differential expression of LipL36. In early-log-phase cultures, LipL36 is one of the most abundant L. kirschneri proteins. However, LipL36 levels drop considerably beginning in mid-log phase. LipL36 expression in vivo was evaluated by examining the humoral immune response to leptospiral antigens in the hamster model of leptospirosis. Hamsters surviving challenge with culture-adapted virulent L. kirschneri generate a strong antibody response to LipL36. In contrast, sera from hamsters surviving challenge with host-adapted L. kirschneri do not recognize LipL36. These findings suggest that LipL36 expression is downregulated during mammalian infection, providing a marker for studying the mechanisms by which pathogenic Leptospira species adapt to the host environment.     

Facial skin mapping: from single point bio‐instrumental evaluation to continuous visualization of skin hydration,barrier function,skin surface pH,and sebum in different ethnic skin types

Dry skin is one of the most important concerns of consumers worldwide. Despite huge efforts over several decades, the personal care industry still does not offer a perfect solution to satisfy the unmet needs of consumers for moisturising treatments in different ethnic groups. The paucity of data for the underlying cellular and biochemical problems in, and the effects of moisturisers on photodamaged facial skin may partly explain this. Mainly, single point measurements are used to understand the effects of products on skin physiology even on surrogate skin sites such as the non‐photodamaged volar forearm. Some groups have developed discontinuous facial maps of skin biophysical properties, however, in 2014 a continuous facial analysis of bio‐instrumental evaluations was developed using a heat map approach. These maps enabled a continuous visualization of features that not only revealed an unexpected complexity of facial skin but also indicated that use of surrogate skin sites for facial skin is inappropriate. We have demonstrated that remarkable gradients of skin hydration, TEWL, skin surface pH and sebum exist within short distances across the face and the gradients are distinctive among different ethnic groups. In addition, these studies have demonstrated that darkly‐pigmented individuals do not necessarily have a better skin barrier function than their less‐pigmented counterparts and that Caucasians have a lower facial skin surface pH compared with more pigmented subjects. Overall, there are no correlations between capacitance, TEWL and skin surface pH including individual topology angle values. Novel 3D camera approaches have also been used to facilitate a more precise assignment of measurement sites and visualisation. The 3D facial colour mappings illustrated precisely the local moisturising effects of a moisturising cream. There were subtle ethnic differences in efficacy that may be related to underlying skin biochemistry and/or ethnic differences in product application. A placebo‐controlled study using conductance measurements in Chinese subjects is also reported. Finally, a new whole face statistical approach has been taken to prove differences in skin parameters but also of moisturiser treatment that adds further to our understanding of the ethnic differences in skin physiology and product application. This paper reviews the background of the development and application of this methodology.     

Haloacetic acid and trihalomethane formation from the chlorination and bromination of aliphatic beta-dicarbonyl acid model compounds

While it is known that resorcinol- and phenol-type aromatic structures within natural organic matter (NOM) react during drinking water chlorination to form trihalomethanes (THMs), limited studies have examined aliphatic-type structures as THM and haloacetic acid (HAA) precursors. A suite of aliphatic acid model compounds were chlorinated and brominated separately in controlled laboratory-scale batch experiments. Four and two beta-dicarbonyl acid compounds were found to be important precursors for the formation of THMs (chloroform and bromoform (71-91% mol/mol)), and dihaloacetic acids (DXAAs) (dichloroacetic acid and dibromoacetic acid (5-68% mol/mol)), respectively, after 24 h at pH 8. Based upon adsorbable organic halide formation, THMs and DXAAs, and to a lesser extent mono and trihaloacetic acids, were the majority (> 80%) of the byproducts produced for most of the aliphatic beta-dicarbonyl acid compounds. Aliphatic beta-diketone-acid-type and beta-keto-acid-type structures could be possible fast- and slow-reacting THM precursors, respectively, and aliphatic beta-keto-acid-type structures are possible slow-reacting DXAA precursors. Aliphatic beta-dicarbonyl acid moieties in natural organic matter, particularly in the hydrophilic fraction, could contribute to the significant formation of THMs and DXAAs observed after chlorination of natural waters.     

Hierarchical growth is necessary and (sometimes) sufficient to self-assemble discrete self-similar fractals

Natural Computing - In this paper, we prove that in the abstract Tile Assembly Model (aTAM), an accretion-based model which only allows for a single tile to attach to a growing assembly at each...     

Vinyl composites—fiberglass reinforced PVC

Glass fiber reinforced PVC has several distinctive and useful properties. Its tensile strength can be twice that of unreinforced PVC. Excellent coupling of the glass fibers to the PVC matrix is required for good retention of tensile strength when exposed to warm water, Its modulus can be twice that of unreinforced PVC and equal to that of wood. PVC's high load carrying capability is not significantly increased to higher temperatures by adding glass fibers as judged by its 264 psi heat deflection temperature. However, by changing the polymeric matrix, glass reinforced vinyl with an increased HDT of 86°C has been produced for higher use temperature. The coefficient of linear thermal expansion of PVC can be cut in half by the addition of glass fibers and has a coefficient equal to that of aluminum. Glass fiber reinforced PVC has exceptionally good resistance to crack propagation and resists shattering as judged by sawing, punching, stapling, and hammering. When properly formulated for weathering resistance, glass fiber reinforced PVC has good color retention, impact retention, and outstanding dimensional stability. As for all glass fiber reinforced plastics, processing equipment must be built for high abrasion resistance for long economical manufacturing runs.     

Gamma linolenic acid attenuates cardiovascular responses to stress in borderline hypertensive rats

The purpose of the present study was to investigate the effects of gamma linolenic acid (GLA) on cardiovascular responses to psychosocial stress (isolation) and to pressor hormones in the genetically borderline hypertensive rat (SHR×WKY). Adult male SHR×WKY were divided into two groups following five weeks of group housing. One group (GLA) received eight weeks constant flow osmotic pumps releasing 0.04 mg GLA in olive oil/kg-hr, while the second group received dummy pumps (DUM). One week following pump implantation, each group was divided into two subgroups and exposed to a four-week experimental period of either continued group housing (no stress) or isolation (stress). A two-week recovery period of group housing followed the experimental period. Blood pressure and heart rate were determined weekly by the tail cuff technique. At the end of the recovery period, animals in the no stress condition were anesthetized and received an arterial cannula for NOR and ANG infusion and direct BP recording. Then the responses to an ED₅₀ of NOR and ANG were determined. All animals were then killed for determination of heart weight and adrenal weight. All groups had mean control period systolic BP values ranging from 143–146 mm Hg. In the no stress condition, neither GLA nor DUM altered BP over the course of the study. However, BP increased in the DUM group durign all four weeks of the isolation period vs the control period (p<0.01), whereas BP increased only in week 1 in the GLA group (p<0.05). Heart rate increased during stress in the DUM group (p<0.05), but not in the GLA group. Vascular reactivity to NOR was unaffected by GLA administration. In contrast, GLA increased the duration of the pressor response to ANG (p<0.01), but tended to decrease the magnitude of the pressor response (p<0.09) vs DUM. GLA had no effect on heart weight/body weight ratio. Adrenal weight/body weight ratio was lower in the DUM/no stress group than all other treatment groups. These data indicate that GLA administration attenuates the cardiovascular responses to chronic stress in animals with a genetic predisposition to hypertension, in the absence of an effect on resting BP. They also demonstrate a limitation of GLA effect, in mature animals, to epigenetic pressor factors. Furthermore, GLA action may involve an alteration of the cardiovascular responses to ANG, but not NOR. These findings suggest that GLA may be useful in preventing the neurogenic triggering of hypertension by chronic stress in genetically stress-sensitive animals.     

Hard prey, soft jaws and the ontogeny of feeding mechanics in the spotted ratfish Hydrolagus colliei.

The spotted ratfish Hydrolagus colliei is a holocephalan fish that consumes hard prey (durophagy) but lacks many morphological characters associated with durophagy in other cartilaginous fishes. We investigated its feeding biomechanics and biting performance to determine whether it can generate bite forces comparable with other durophagous elasmobranchs, how biting performance changes over ontogeny (21-44 cm SL) and whether biomechanical modelling can accurately predict feeding performance in holocephalans. Hydrolagus colliei can generate absolute and mass-specific bite forces comparable with other durophagous elasmobranchs (anterior=104 N, posterior=191 N) and has the highest jaw leverage of any cartilaginous fish studied. Modelling indicated that cranial geometry stabilizes the jaw joint by equitably distributing forces throughout the feeding mechanism and that positive allometry of bite force is due to hyperallometric mechanical advantage. However, bite forces measured through tetanic stimulation of the adductor musculature increased isometrically. The jaw adductors of H. colliei fatigued more rapidly than those of the piscivorous spiny dogfish Squalus acanthias as well. The feeding mechanism of H. colliei is a volume-constrained system in which negative allometry of cranial dimensions leaves relatively less room for musculature. Jaw adductor force, however, is maintained through ontogenetic changes in muscle architecture.     

Automatic correction of level set based subvoxel precise centerlines for virtual colonoscopy using the colon outer wall

Virtual colonoscopy (VC) is becoming a more prevalent method to detect and diagnose colorectal cancer. An essential component of using VC to detect cancerous polyps, especially in conjunction with computer-aided diagnosis, is the accurate calculation of the centerline of the colon. While the colon is often modeled as a simple cylinder, the amount of colonic distention may vary between patients and within the same patient often causing loops and multiple disconnected segments to be present in the colon segmentation. These variations have caused previous centerline algorithms to fail to capture a complete and accurate centerline for all colons. We have developed an automatic method to determine from a computed tomography (CT) VC a subvoxel precise centerline that is accurate even in cases of over-distended or under-distended colons. In this algorithm, the loops in the colon caused by over-distention are detected and removed when the centerline calculation is performed. Also, a newly developed method for the detection and segmentation of the outer wall of the colon is used to connect collapsed portions of the colon where the lumen segmentation fails to produce a continuous centerline. These two methods allow for a complete and accurate centerline to be calculated in uniformly distended colons as well as in colons containing segments which are over-distended and/or under-distended. We have demonstrated successfully the effectiveness of our algorithm on 50 cases, 25 of which resulted in erroneous solutions by previous centerline algorithms due to variability in the colon distention.     

Expression and distribution of leptospiral outer membrane components during renal infection of hamsters

The outer membrane of pathogenic Leptospira species grown in culture media contains lipopolysaccharide (LPS), a porin (OmpL1), and several lipoproteins, including LipL36 and LipL41. The purpose of this study was to characterize the expression and distribution of these outer membrane antigens during renal infection. Hamsters were challenged with host-derived Leptospira kirschneri to generate sera which contained antibodies to antigens expressed in vivo. Immunoblotting performed with sera from animals challenged with these host-derived organisms demonstrated reactivity with OmpL1, LipL41, and several other proteins but not with LipL36. Although LipL36 is a prominent outer membrane antigen of cultivated L. kirschneri, its expression also could not be detected in infected hamster kidney tissue by immunohistochemistry, indicating that expression of this protein is down-regulated in vivo. In contrast, LPS, OmpL1, and LipL41 were demonstrated on organisms colonizing the lumen of proximal convoluted renal tubules at both 10 and 28 days postinfection. Tubular epithelial cells around the luminal colonies had fine granular cytoplasmic LPS. When the cellular inflammatory response was present in the renal interstitium at 28 days postinfection, LPS and OmpL1 were also detectable within interstitial phagocytes. These data establish that outer membrane components expressed during infection have roles in the induction and persistence of leptospiral interstitial nephritis.     

Effect of diesel soot contaminated oil on engine wear — investigation of novel oil formulations

Exhaust Gas Re-circulation (EGR) has been found to be very effective in reducing emissions of oxides of nitrogen, for light duty diesel engines. However, EGR results in a sharp increase in particulate matter emissions in heavy-duty diesel engines.The effects of soot contaminated engine oil on wear of engine components was examined using a statistically designed experiment. The three oil properties studied were phosphorous level, dispersant level and sulfonate substrate level. The above three variables were formulated at two levels: High (1) and Low (−1). This resulted in a 2³ matrix (eight oil blends). The effect of soot was also taken into consideration, which resulted in a 2⁴ factorial experiment.A three-body wear machine was designed and developed to simulate and estimate the extent of wear. Ball-on-flat-disk tests were conducted to qualitatively study wear by comparing wear scars due to soot with wear scars due to a known abrasive (alumina). A Scanning Electron Microscope (SEM) was used to study the microstructures of the wear scars. Surface chemical analysis was performed on soot particles and wear scars using Energy–Dispersive X-ray Analysis (EDAX).Results show that diesel soot interacts with oil additives reducing the oil's anti-wear properties possibly by abrasive wear mechanism. Statistical analysis (GLM) showed that the phosphorous level plays a dominant role on oil's wear performance. The effect of dispersant level was not very significant, though on an average, higher dispersant levels reduced wear. The effect of sulfonate was not revealed within the range of these concentrations. Ball-on-flat-disk type tests also revealed the increased wear due to the presence of soot. SEM studies of Wear Scar Diameters suggest that soot is abrasive.