bluuebunny/crossref_metadata_2025 huggingface dataset:
New tobacco trends: Waterpipe (hookah) smoking and implications for healthcare providers
Authors: Noonan Devon , Kulbok Pamela A.
Published: May 2009

Purpose: To inform healthcare providers about waterpipe smoking, a new trend in tobacco use that is gaining popularity among adolescents and young adults.

Data sources: American Lung Association Tobacco Policy Alert on Waterpipe Smoking, World Health Organization Tobacco Regulation Advisory Note on Waterpipe Smoking, and pertinent publications available in the literature.

Conclusions: Waterpipe smoking is a new trend in tobacco use that is associated with multiple health problems, including addiction. Healthcare providers should be aware of new tobacco trends that may affect patients, such as waterpipe smoking, that are potential gateways to nicotine addiction.

Implications for practice: Tobacco comes in many forms, all of which are addicting. Healthcare providers must be knowledgeable about new forms of tobacco to address all types of tobacco use with patients. Healthcare providers also have a responsibility to educate patients about the health risks inherent in these products to help prevent the long‐lasting problem of nicotine addiction.
Influence of angiotensin‐converting enzyme I/D gene polymorphism on clinical and histological correlates of chronic hepatitis C
Authors: Fabris Carlo , Smirne Carlo , Fangazio Stefano , Toniutto Pierluigi , Burlone Michela , Minisini Rosalba , Bitetto Davide , Falleti Edmondo , Cerutti Andrea , Pirisi Mario
Published: April 2009

Aim:  This study aimed to verify the relationship between the insertion–deletion (I/D) polymorphism of angiotensin‐converting enzyme (ACE) and clinical and histological correlates of chronic hepatitis C.

Methods:  Two‐hundred and fifty‐eight, treatment naive, unselected hepatitis C virus (HCV) RNA‐positive patients and 210 controls were studied. ACE allelic variants were determined by polymerase chain reaction.

Results:  Mean staging scores adjusted for age, body mass index (BMI) and alcohol consumption were: men, D/* = 2.283; men, I/I = 2.092; women, D/* = 2.241; and women, I/I = 3.283 (P = 0.028). Age‐adjusted mean BMI were: men, D/* = 25.01; men, I/I = 24.87; women, D/* = 23.73; and women, I/I = 22.50 (P = 0.006). Age and BMI‐adjusted mean low‐density lipoprotein (LDL)/ high‐density lipoprotein (HDL) cholesterol ratios were: men, D/* = 2.344; men, I/I = 2.283; women, D/* = 1.916; and women, I/I = 1.903 (P = 0.004). Histological grading correlated positively with triglycerides and negatively with HDL and LDL cholesterol (P < 0.0001).

Conclusion:  Female ACE I/I homozygotes have higher liver fibrosis scores in comparison to D/* women and to men; moreover, they are leaner and have a lower LDL/HDL cholesterol ratio. These observations suggest a possible mutual influence between ACE polymorphism, serum lipid concentrations and outcome of chronic HCV infection.
TRPV1 channels control synaptic plasticity in the developing superior colliculus
Authors: Maione Sabatino , Cristino Luigia , Migliozzi Anna Lucia , Georgiou Anne L. , Starowicz Katarzyna , Salt Thomas E. , Di Marzo Vincenzo
Published: May 2009

Long‐term depression (LTD) in the rodent superior colliculus (SC) is regarded as a model of synaptic refinement because it can be induced during development but not in adults. We investigated the role of transient receptor potential vanilloid type‐1 (TRPV1) channels in this type of synaptic plasticity. Experiments were carried out in pigmented mice aged between postnatal day 8 (P8) and 42 (P42) and in adult mice. Retinal axons to the SC were labelled by injection of cholera toxin‐β (CTβ) into the eye. Immunohistochemical staining for CTβ, TRPV1 and markers of glutamatergic and GABAergic cells and fibres (VGLUT1 and VGAT or GAD65, respectively) was performed by using multiple immunofluorescence. This showed that both glutamatergic retinal afferents to, and some GABAergic neurones in, the superficial SC are TRPV1 positive in juvenile but not adult mice. Field potential recordings were made from the superficial grey layer in parasagittal SC slices, and LTD (76 ± 8% of control responses) was induced with a 50 Hz, 20 s tetanus. Activation of TRPV1 with resiniferatoxin also reduced field potential amplitude to 84 ± 8% of control values. Blockade of TRPV1 with the selective antagonist 5′‐iodo‐resiniferatoxin prevented the induction of LTD (98 ± 4% of control values), but did not cause its reversal if LTD was already established. N‐acylphosphatidylethanolamine‐specific phospholipase D and 12‐lipoxygenase, two proposed endovanilloid biosynthesizing enzymes, were co‐expressed with TRPV1 in the SC at P14 and P28. These results suggest that TRPV1 modulates retinocollicular responses in the developing SC and is activated during tetanic stimulation by endovanilloid ligands to participate in the induction of LTD.
Superconductivity at the Two-Dimensional Limit
Authors: Qin Shengyong , Kim Jungdae , Niu Qian , Shih Chih-Kang
Published: May 2009

Superconducting Limits

An electron wave function extends in three dimensions. Pairing up into Cooper pairs and reducing the temperatures can induce superconductivity in a material. How robust is an inherently three-dimensional effect when the dimensions are restricted?

Qin
et al.

(p.
1314
, published online 30 April) looked at the thickness dependence of superconductivity in thin lead films. In contrast to earlier, less direct measurements, superconductivity was robust, even down to films of only two monolayers.
Lipopolysaccharide-Induced Biliary Factors Enhance Invasion ofSalmonella enteritidisin a Rat Model
Authors: Islam Abul F. M. W. , Moss Nathan D. , Dai Yung , Smith Murray S. R. , Collins Andrew M. , Jackson Graham D. F.
Published: May 2009

In this study, the role of the hepatobiliary system in the early pathogenesis ofSalmonella enteritidisinfection was investigated in a rat model. Intravenous (i.v.) challenge with lipopolysaccharide (LPS) has previously been shown to enhance the translocation of normal gut flora. We first confirmed that LPS can similarly promote the invasion ofS. enteritidis. Oral infection of outbred Australian Albino Wistar rats with 106to 107CFU ofS. enteritidisled to widespread tissue invasion after days. If animals were similarly challenged after intravenous administration ofS. enteritidisLPS (3 to 900 μg/kg of body weight), significant invasion of the livers and mesenteric lymph nodes (MLN) occurred within 24 h, with invasion of the liver increasing in a dose-dependent fashion (P< 0.01). If bile was prevented from reaching the intestine by bile duct ligation or cannulation, bacterial invasion of the liver and MLN was almost totally abrogated (P< 0.001). As i.v. challenge with LPS could induce the delivery of inflammatory mediators into the bile, biliary tumor necrosis factor alpha (TNF-α) concentrations were measured by bioassay. Biliary concentrations of TNF-α rose shortly after LPS challenge, peaked with a mean concentration of 27.0 ng/ml at around 1 h postchallenge, and returned to baseline levels (3.1 ng/ml) after 2.5 h. Although TNF-α cannot be directly implicated in the invasion process, we conclude that the invasiveness of the enteric pathogenS. enteritidisis enhanced by the presence of LPS in the blood and that this enhanced invasion is at least in part a consequence of the delivery of inflammatory mediators to the gastrointestinal tract by the hepatobiliary system.
Strain-Specific Restriction of the Antiphagocytic Property of Group A Streptococcal M Proteins
Authors: Kotarsky Heike , Thern Anette , Lindahl Gunnar , Sjöbring Ulf
Published: May 2009

Group A streptococcal M proteins are type-specific virulence factors that inhibit phagocytosis. We used two M proteins, M5 and Emm22, to analyze the influence of genetic background on the properties of M proteins. Mutant strains, engineered to lack these M proteins, were complemented with genes encoding the homologous or heterologous M protein, and the complemented strains were analyzed for phagocytosis resistance. Neither the M5 nor the Emm22 protein conferred phagocytosis resistance in the heterologous background, but they did do so in the homologous background. This was not due to lack of surface expression in the heterologous background. Moreover, the M5 and Emm22 proteins expressed in heterologous background appeared to have normal structure, since they were not affected in their ability to bind different human plasma proteins. In particular, M5 or Emm22 had normal ability to bind human complement inhibitors, a property that has been implicated in phagocytosis resistance. Results similar to those obtained with M5 and Emm22 were obtained in experiments with the M6 and Emm4 proteins. Together, these data suggest that the surface expression of M protein alone may not be sufficient to confer phagocytosis resistance and consequently that strain-specific factors other than M and Emm proteins may contribute to the ability of group A streptococci to resist phagocytosis.
Two Distinct Antigenic Types of the Polysaccharide Chains of
Helicobacter pylori
Lipopolysaccharides Characterized by Reactivity with Sera from Humans with Natural Infection
Authors: Yokota Shin-Ichi , Amano Ken-Ichi , Shibata Yoshiko , Nakajima Mizuho , Suzuki Miyuki , Hayashi Shunji , Fujii Nobuhiro , Yokochi Takashi
Published: May 2009

We have purified lipopolysaccharides (LPS) from 10
Helicobacter pylori
clinical isolates which were selected on the basis of chemotype and antigenic variation. Data from immunoblotting of the purified LPS with sera from humans with
H. pylori
infection and from absorption of the sera with LPS indicated the presence of two distinct epitopes, termed the highly antigenic and the weakly antigenic epitopes, on the polysaccharide chains. Among 68
H. pylori
clinical isolates, all smooth strains possessed either epitope; the epitopes were each carried by about 50% of the smooth strains. Thus,
H. pylori
strains can be classified into three types on the basis of their antigenicity in humans: those with smooth LPS carrying the highly antigenic epitope, those with smooth LPS carrying the weakly antigenic epitope, and those with rough LPS. Sera from humans with
H. pylori
infection could be grouped into three categories: those containing immunoglobulin G (IgG) antibodies against the highly antigenic epitope, those containing IgG against the weakly antigenic epitope, and those containing both specific IgGs; these groups made up about 50%, less than 10%, and about 40%, respectively, of all infected sera tested. In other words, IgG against the highly antigenic epitope were detected in more than 90% of
H. pylori
-infected individuals with high titers. IgG against the weakly antigenic epitope were detected in about 50% of the sera tested; however, the antibody titers were low. The two human epitopes existed independently from the mimic structures of Lewis antigens, which are known to be an important epitope of
H. pylori
LPS. No significant relationship between the reactivities toward purified LPS of human sera and a panel of anti-Lewis antigen antibodies was found. Moreover, the reactivities of the anti-Lewis antigen antibodies, but not human sera, were sensitive to particular α-
l
-fucosidases. The human epitopes appeared to be located on O-polysaccharide chains containing endo-β-galactosidase-sensitive galactose residues as the backbone. Data from chemical analyses indicated that all LPS commonly contained galactose, glucosamine, glucose, and fucose (except one rough strain) as probable polysaccharide components, together with typical components of inner core and lipid A. We were not able to distinguish between the differences of antigenicity in humans by on the basis of the chemical composition of the LPS.
Contribution of
Escherichia coli
Alpha-Hemolysin to Bacterial Virulence and to Intraperitoneal Alterations in Peritonitis
Authors: May Addison K. , Gleason Thomas G. , Sawyer Robert G. , Pruett Timothy L.
Published: May 2009

Alpha-hemolysin (Hly) is a common exotoxin produced by
Escherichia coli
that enhances virulence in a number of clinical infections. The addition of hemolysin production to laboratory bacterial strains is known to increase the lethality of
E. coli
peritonitis. However, the mechanisms involved have not been determined and the contribution of hemolysin to the alterations in the host intraperitoneal environment and the leukocyte response is not known. Utilizing a rat peritonitis model, we show that wild-type hemolytic
E. coli
strains have a significant competitive advantage over nonhemolytic strains within the peritoneum. To examine the specific contribution of Hly to
E. coli
-induced virulence and alterations within the peritoneum, a mixed peritonitis model of
E. coli
,
Bacteroides fragilis
, and sterile fecal adjuvant was used. Three transformed
E. coli
strains were utilized: one strongly secretes active hemolysin (WAF 270), a second secretes active hemolysin but a reduced amount (WAF 260), and the third does not produce hemolysin (WAF 108). After an equal inoculum of each of the three strains, WAF 270 produced a markedly increased lethality and an increased recovery of both
E. coli
and
B. fragilis
from the host relative to the other strains. Changes in the intraperitoneal pH, degree of erythrocyte lysis, and recruitment and viability of leukocytes within the peritoneum following the induction of peritonitis differed significantly between the strongly hemolytic and nonhemolytic strains. Induction of peritonitis with WAF 270 caused a pronounced decrease in intraperitoneal pH, lysis of most of the intraperitoneal erythrocytes, and a marked decrease in recoverable viable leukocytes compared to WAF 108. Thus, hemolysin production by
E. coli
within the peritoneum may alter not only the host's ability to control the hemolytic strain itself but also other organisms.
Subclinical Chlamydial Infection of the Female Mouse Genital Tract Generates a Potent Protective Immune Response: Implications for Development of Live Attenuated Chlamydial Vaccine Strains
Authors: Su Hua , Messer Ronald , Whitmire William , Hughes Scott , Caldwell Harlan D.
Published: May 2009

Chlamydia trachomatisis a major cause of sexually transmitted disease (STD) for which a vaccine is needed. CD4+T-helper type 1 (Th1) cell-mediated immunity is an important component of protective immunity against murine chlamydial genital infection. Conventional vaccine approaches have not proven effective in eliciting chlamydial-specific CD4 Th1 immunity at the genital mucosa. Thus, it is possible that the development of a highly efficacious vaccine against genital infection will depend on the generation of a live attenuatedC. trachomatisvaccine. Attenuated strains ofC. trachomatisdo not exist, so their potential utility as vaccines cannot be tested in animal models of infection. We have developed a surrogate model to study the effect of chlamydial attenuation on infection and immunity of the female genital tract by treating mice with a subchlamydiacidal concentration of oxytetracycline following vaginal infection. Compared to untreated control mice, antibiotic-treated mice shed significantly fewer infectious organisms (3 log10) from the cervico-vagina, produced a minimal inflammatory response in urogenital tissue, and did not experience infection-related sequelae. Antibiotic-treated mice generated levels of chlamydia-specific antibody and cell-mediated immunity equivalent to those of control mice. Importantly, antibiotic-treated mice were found to be as immune as control untreated mice when rechallenged vaginally. These findings demonstrate that subclinical chlamydial infection of the murine female genital tract is sufficient to stimulate a potent protective immune response. They also present indirect evidence supporting the possible use of live attenuated chlamydial organisms in the development of vaccines against chlamydial STDs.
Comparative Evaluation of Low-Molecular-Mass Proteins fromMycobacterium tuberculosisIdentifies Members of the ESAT-6 Family as Immunodominant T-Cell Antigens
Authors: Skjøt Rikke Louise Vinther , Oettinger Thomas , Rosenkrands Ida , Ravn Pernille , Brock Inger , Jacobsen Susanne , Andersen Peter
Published: May 2009

Culture filtrate fromMycobacterium tuberculosiscontains protective antigens of relevance for the generation of a new antituberculosis vaccine. We have identified two previously uncharacterizedM. tuberculosisproteins (TB7.3 and TB10.4) from the highly active low-mass fraction of culture filtrate. The molecules were characterized, mapped in a two-dimensional electrophoresis reference map of short-term culture filtrate, and compared with another recently identified low-mass protein, CFP10 (F. X. Berthet, P. B. Rasmussen, I. Rosenkrands, P. Andersen, and B. Gicquel. Microbiology 144:3195–3203, 1998), and the well-described ESAT-6 antigen. Genetic analyses demonstrated that TB10.4 as well as CFP10 belongs to the ESAT-6 family of low-mass proteins, whereas TB7.3 is a low-molecular-mass protein outside this family. The proteins were expressed inEscherichia coli, and their immunogenicity was tested in cultures of peripheral blood mononuclear cells from human tuberculosis (TB) patients,Mycobacterium bovisBCG-vaccinated donors, and nonvaccinated donors. The two ESAT-6 family members, TB10.4 and CFP10, were very strongly recognized and induced gamma interferon release at the same level (CFP10) as or at an even higher level (TB10.4) than ESAT-6. The non-ESAT-6 family member, TB7.3, for comparison, was recognized at a much lower level. CFP10 was found to distinguish TB patients from BCG-vaccinated donors and is, together with ESAT-6, an interesting candidate for the diagnosis of TB. The striking immunodominance of antigens within the ESAT-6 family is discussed, and hypotheses are presented to explain this targeting of the immune response during TB infection.
Intracellular and Extracellular Cytokine Production by Human Mixed Mononuclear Cells in Response to Group B Streptococci
Authors: Kwak Daniel J. , Augustine Nancy H. , Borges Wellington G. , Joyner Joanna L. , Green Wayne F. , Hill Harry R.
Published: May 2009

Group B streptococci (GBS) are a major cause of severe infection in newborns, pregnant females, and other immunocompromised hosts. Infection often includes septicemia, shock, pneumonia, and respiratory failure. In previous studies, we have reported that GBS induce marked production of tumor necrosis factor alpha (TNF-α) by human mononuclear cells. The present study was designed to measure the production of TNF-α as well as additional cytokines, including interleukin 1β (IL-1β), IL-6, IL-8, IL-12, and gamma interferon (IFN-γ) but also to determine from what cells and at what time point during incubation with GBS that these cytokines are produced. Mixed mononuclear cells were incubated with heat-killed GBS, media alone, or 1 μg ofEscherichia colilipopolysaccharide (LPS). Brefeldin A was added to each sample prior to staining, which prevented the export of cytokines by the Golgi apparatus. The cells were then stained with the appropriate conjugated antibodies and analyzed by using a flow cytometer. Results indicate that intracellular cytokines appear, in almost all cases, simultaneous to or before secreted proteins are detected. In contrast to the response to LPS, where TNF-α, IL-1β, IL-6, and IL-8 appear almost simultaneously, the human monocyte response to GBS results in the production of TNF-α but delayed appearance of IL-1β, IL-6, and IL-8. The lymphocyte response to GBS was also strikingly different from that to LPS in that both secreted IFN-γ and IL-12 was detected, while LPS failed to induce production of these critical cytokines. This suggests an important role for TNF-α, IFN-γ, and IL-12 in GBS pathogenesis and/or immunity.
Depletion of CD8+T Cells In Vivo Impairs Host Defense of Mice Resistant and Susceptible to Pulmonary Paracoccidioidomycosis
Authors: Cano Luz E. , Singer-Vermes Lúcia M. , Costa Tania A. , Mengel José O. , Xidieh Cynthia F. , Arruda Celina , André Denise C. , Vaz Celidéia A. C. , Burger Eva , Calich Vera L. G.
Published: May 2009

Using a pulmonary model of infection, we demonstrated previously that A/Sn and B10.A mice are, respectively, resistant and susceptible toParacoccidioides brasiliensisinfection. Employing the same experimental model, we examined herein the role of CD8+T cells in the course of paracoccidioidomycosis. Treatment with anti-CD8 monoclonal antibodies caused a selective depletion of pulmonary and splenic CD8+T cells in both mouse strains. The number of pulmonary CD4+T cells and immunoglobulin-positive cells was independent of the number of CD8+T cells. In susceptible mice, the loss of CD8+T cells by in vivo treatment with anti-CD8 monoclonal antibodies impaired the clearance of yeasts from the lungs and increased the fungal dissemination to the liver and spleen. The same treatment in resistant mice increased fungal dissemination to extrapulmonary tissues but did not alter the pulmonary fungal load. Furthermore, CD8+T-cell depletion did not modify delayed-type hypersensitivity reactions of A/Sn mice but increased these reactions in B10.A mice. The production ofP. brasiliensis-specific antibodies by resistant and susceptible mice depleted of CD8+T cells was similar to that of mice given control antibody. Histopathologically, depletion of CD8+T cells did not disorganize the focal granulomatous lesions developed by both mouse strains. These results indicate that CD8+T cells are necessary for optimal clearance of the fungus from tissues of mice infected withP. brasiliensisand demonstrate more prominent protective activity by those cells in the immune responses mounted by susceptible animals.
Interaction of Enteropathogenic or Enterohemorrhagic
Escherichia coli
with HeLa Cells Results in Translocation of Cortactin to the Bacterial Adherence Site
Authors: Cantarelli Vlademir V. , Takahashi Akira , Akeda Yukihiro , Nagayama Kenichi , Honda Takeshi
Published: May 2009

Infection of cultured HeLa epithelial cells with enteropathogenic
Escherichia coli
(EPEC) or enterohemorrhagic
E. coli
(EHEC) O157:H7 results in accumulation of cortactin under the adherent bacteria. Tyrosine phosphorylation of cortactin is not induced following HeLa cell infection with EHEC or EPEC, contrary to what has been reported to occur with
Shigella flexneri.
Plasmodium falciparum
Rosette Formation Is Uncommon in Isolates from Pregnant Women
Authors: Rogerson Stephen J. , Beeson James G. , Mhango Chisale G. , Dzinjalamala Fraction K. , Molyneux Malcolm E.
Published: May 2009

We examined the formation of
Plasmodium falciparum
erythrocyte rosettes using parasite isolates from placental or peripheral blood of pregnant Malawian women and from peripheral blood of children. Five of 23 placental isolates, 23 of 38 maternal peripheral isolates, and 136 of 139 child peripheral isolates formed rosettes. Placental isolates formed fewer rosettes than maternal isolates (range, 0 to 7.5% versus 0 to 33.5%;
P
= 0.002), and both formed fewer rosettes than isolates cultured from children (range, 0 to 56%;
P
< 0.0001). Rosette formation is common in infections of children but uncommon in pregnancy and rarely detected in placental isolates.
High Levels of Inducible Nitric Oxide Synthase mRNA Are Associated with Increased Monocyte Counts in Blood and Have a Beneficial Role inPlasmodium falciparumMalaria
Authors: Chiwakata Collins Batsirai , Hemmer Christoph Josef , Dietrich Manfred
Published: May 2009

To date, there have been conflicting reports concerning the clinical significance of nitric oxide (NO) inPlasmodium falciparummalaria. Some authors have proposed that NO contributes to the development of severe and complicated malaria, while others have argued that NO has a protective role. To investigate these apparently contradictory reports, reverse transcription-coupled PCR was used to study inducible NO synthase (iNOS) in whole-blood RNA samples from patients with severe and complicated malaria or uncomplicated malaria and from healthy donors. This work produced three principal findings. First, samples of patients with severe and complicated malaria were variably positive, with weak to moderate intensity. Markedly higher iNOS RNA levels were observed in samples of patients with uncomplicated malaria than in patients with severe and complicated malaria. Samples of healthy donors were uniformly negative. Second, since we initially demonstrated iNOS expression in whole-blood RNA samples, we extended our investigations to individual blood cells such as monocytes, lymphocytes, neutrophils, and platelets to identify the cellular source of iNOS. We found that iNOS was expressed predominantly in monocytes. Third, retrospective statistical analysis of monocyte counts clearly demonstrated that patients with uncomplicated malaria had higher monocyte counts at the time of presentation than patients with severe and complicated malaria. Taken together, our findings give room to the interpretation that NO may have a beneficial rather than a deleterious role in falciparum malaria.
Archaeosome Vaccine Adjuvants Induce Strong Humoral, Cell-Mediated, and Memory Responses: Comparison to Conventional Liposomes and Alum
Authors: Krishnan Lakshmi , Dicaire Chantal J. , Patel Girishchandra B. , Sprott G. Dennis
Published: May 2009

Ether glycerolipids extracted from various archaeobacteria were formulated into liposomes (archaeosomes) possessing strong adjuvant properties. Mice of varying genetic backgrounds, immunized by different parenteral routes with bovine serum albumin (BSA) entrapped in archaeosomes (∼200-nm vesicles), demonstrated markedly enhanced serum anti-BSA antibody titers. These titers were often comparable to those achieved with Freund's adjuvant and considerably more than those with alum or conventional liposomes (phosphatidylcholine-phosphatidylglycerol-cholesterol, 1.8:0.2:1.5 molar ratio). Furthermore, antigen-specific immunoglobulin G1 (IgG1), IgG2a, and IgG2b isotype antibodies were all induced. Association of BSA with the lipid vesicles was required for induction of a strong response, and >80% of the protein was internalized within most archaeosome types, suggesting efficient release of antigen in vivo. Encapsulation of ovalbumin and hen egg lysozyme within archaeosomes showed similar immune responses. Antigen-archaeosome immunizations also induced a strong cell-mediated immune response: antigen-dependent proliferation and substantial production of cytokines gamma interferon (Th1) and interleukin-4 (IL-4) (Th2) by spleen cells in vitro. In contrast, conventional liposomes induced little cell-mediated immunity, whereas alum stimulated only an IL-4 response. In contrast to alum and Freund's adjuvant, archaeosomes composed ofThermoplasma acidophilumlipids evoked a dramatic memory antibody response to the encapsulated protein (at ∼300 days) after only two initial immunizations (days 0 and 14). This correlated with increased antigen-specific cell cycling of CD4+T cells: increase in synthetic (S) and mitotic (G2/M) and decrease in resting (G1) phases. Thus, archaeosomes may be potent vaccine carriers capable of facilitating strong primary and memory humoral, and cell-mediated immune responses to the entrapped antigen.
Perturbations in Eosinophil Homeostasis following Treatment of Lymphatic Filariasis
Authors: Gopinath Ramya , Hanna L. E. , Kumaraswami V. , Perumal V. , Kavitha V. , Vijayasekaran V. , Nutman Thomas B.
Published: May 2009

Treatment of patients with patentWuchereria bancroftiinfection results in an acute clinical reaction and peripheral eosinophilia. To investigate the dynamics of the eosinophil response, changes in eosinophil activation and degranulation and plasma levels of eosinophil-active chemokines and cytokines were studied in 15 microfilaremic individuals in south India by sequential blood sampling before and after administration of 300 mg of diethylcarbamazine (DEC). Clinical symptoms occurred within 24 h. Plasma interleukin-5 (IL-5) and RANTES levels peaked 1 to 2 days posttreatment, preceding a peak peripheral eosinophil count at day 4. Major basic protein secretion from eosinophils paralleled IL-5 secretion, while levels of eosinophil-derived neurotoxin peaked at day 13 after treatment. Expression of the activation markers HLA-DR and CD25 on eosinophils rose markedly immediately after treatment, while expression of VLA-4 and α4β7 showed an early peak within 24 h and a second peak at day 13. Thus, the posttreatment reactions seen in filarial infections can be divided into an early phase with killing of microfilariae, clinical symptomatology, increases in plasma IL-5 and RANTES levels, and eosinophil activation and degranulation and a later phase with expression of surface integrins on eosinophils, recruitment of eosinophils from the bone marrow to tissues, and clearance of parasite antigen.
X-ray absorption spectroscopy of aqueous solutions in the X-ray energy region from 1800 to 3700 eV
Authors: Jürgensen Astrid
Published: May 2009

A methodology for the X-ray XANES and EXAFS analyses of aqueous solutions was developed for the Double Crystal Monochromator (DCM) beamline of the Canadian Synchrotron Radiation Facility (CSRF). The technique is based on the implementation of a liquid cell, which is compatible with the standard solid-state sample holding mechanism used at the beamlines of CSRF and the Canadian Light Source (CLS). Aluminized Mylar film with a thickness of 5 μm has been thoroughly investigated for use as a window material for the cell, and found acceptable for the collection of the S, Cl, and K K-edge spectra of ions in aqueous solution, but not for P K-edge or Rb L2,3-edge spectra. Some of the limitations of this methodology, such as diffusion of the solution through the window and the detection limit at low ion concentration, are investigated and discussed.
Matrix effects on mass spectrometric determinations of four pharmaceuticals and personal care products in water, sediments, and biota
Authors: Dussault Ève B. , Balakrishnan Vimal K. , Solomon Keith R. , Sibley Paul K.
Published: May 2009

Simple analytical methods were developed for the extraction and determination of four pharmaceuticals and personal care products (PPCPs) from water, sediments, and biota. PPCPs were determined using tandem LC–MS in electrospray ionization mode, and interactions with matrix co-eluents were investigated. Extractions of water samples were performed using solid-phase extraction (SPE), sediments were extracted by pressurized liquid extraction (PLE), and biota was extracted by liquid extraction. The selected analytical methods yielded recoveries ≥ 61% in all matrixes. Matrix interactions were investigated throughout the linear range of quantification of each compound, revealing that dissolved salts had relatively minor effects on ionization (between 14% suppression to 12% enhancement), but that sediment and biota extracts caused significant matrix effects (ranging from 56% suppression to 25% enhancement). The direction and magnitude of matrix interactions reflected the physico-chemical properties of each analyte, particularly their pKa. Among the compounds analyzed in electrospray positive mode, carbamazepine was insensitive to matrix interactions, because it is a strong proton acceptor (pKa= 14.0). In contrast, atorvastatin (pKa= 4.5), a weaker proton acceptor, was particularly sensitive to matrix effects. For those compounds analyzed in negative-ion mode, sample alkalinity was found to be important. With a pKaof 10.4, 17α-ethinylestradiol generally exhibited matrix enhancement with increased sample alkalinity. However, the presence of acidic co-eluents contributed to matrix suppression. Lastly, TCS was particularly sensitive to matrix suppression, as its circumneutral pKa(7.9) caused even slight changes in sample pH to considerably impact ionization. We conclude that while different matrixes have clear impacts on ionization of these PPCPs, matrix effects can be quantified and overcome.
On the reactivity of indium(III) benzenechalcogenolates (chalcogen = sulfur and selenium) towards organyl halides for the synthesis of organyl phenyl chalcogenides
Authors: Peppe Clovis , Borges de Castro Liérson
Published: May 2009

The reactivity of indium(III) benzenechalcogenolates (chalcogen = sulfur, selenium) towards organyl halides (organyl = alkyl, allyl, benzyl, acyl) was examined. A practical one-pot method to prepare organyl phenyl chalcogenides from indium metal and diphenyl dichalcogenide was found. The coupling is fairly broad in scope and generally works better for organyl halides capable to produce stable carbocations.
Hyperoxia impairs postnatal alveolar epithelial development via NADPH oxidase in newborn mice
Authors: Auten Richard L. , Mason S. Nicholas , Auten Kathryn M. , Brahmajothi Mulugu
Published: May 2009

Hyperoxia disrupts postnatal lung development in part through inducing inflammation. To determine the contribution of leukocyte-derived reactive oxygen species, we exposed newborn wild-type and NADPH oxidase p47 phox subunit null (p47 phox−/−) mice to air or acute hyperoxia (95% O2) for up to 11 days. Hyperoxia-induced pulmonary neutrophil influx was similar in wild-type and p47−/− mice at postnatal days (P) 7 and 11. Macrophages were decreased in wild-type hyperoxia-exposed mice compared with p47 phox−/− mice at P11. Hyperoxia impaired type II alveolar epithelial cell and bronchiolar epithelial cell proliferation, but depression of type II cell proliferation was significantly less in p47−/− mice at P3 and P7, when inflammation was minimal. We found reciprocal results for the expression of the cell cycle inhibitor p21 cip/waf in type II cells, which was induced in 95% O2-exposed wild-type mice, but significantly less in p47 phox−/− littermates at P7. Despite partial preservation of type II cell proliferation, deletion of p47 phox did not prevent the major adverse effects of hyperoxia on alveolar development estimated by morphometry at P11, but hyperoxia impairment of elastin deposition at alveolar septal crests was significantly worse in wild-type vs. p47 phox−/− mice at P11. Since we found that p47 phox is expressed in a subset of alveolar epithelial cells, its deletion may protect postnatal type II alveolar epithelial proliferation from hyperoxia through effects on epithelial as well as phagocyte-generated superoxide.
Knockdown of ERp57 increases BiP/GRP78 induction and protects against hyperoxia and tunicamycin-induced apoptosis
Authors: Xu Dong , Perez Ricardo E. , Rezaiekhaligh Mohammad H. , Bourdi Mohammed , Truog William E.
Published: May 2009

Supplemental oxygen therapy (hyperoxia) in preterm babies with respiratory stress is associated with lung injury and the development of bronchopulmonary dysplasia. Endoplasmic reticulum (ER) homeostasis plays critical roles in maintaining cellular functions such as protein synthesis, folding, and secretion. Interruption of ER homeostasis causes ER stress and triggers the unfolded protein response, which can lead to apoptosis in persistently stressed cells. ERp57 is an ER protein and is associated with calreticulin and calnexin in protein glycosylation. In this study, we found hyperoxia downregulated ERp57 in neonatal rat lungs and cultured human endothelial cells. Transient transfection of ERp57 small interfering RNA significantly knocked down ERp57 expression and reduced hyperoxia- or tunicamycin-induced apoptosis in human endothelial cells. Apoptosis was decreased from 26.8 to 9.9% in hyperoxia-exposed cells and from 37.8 to 5.0% in tunicamycin-treated cells. The activation of caspase-3 induced by hyperoxia or tunicamycin was diminished and immunoglobulin heavy chain-binding protein/glucose-regulated protein 78-kDa (BiP/GRP78) induction was increased in ERp57 knockdown cells. Overexpression of ERp57 exacerbated hyperoxia- or tunicamycin-induced apoptosis in human endothelial cells. Apoptosis was increased from 10.1 to 14.3% in hyperoxia-exposed cells and from 14.0 to 21.2% in tunicamycin-treated cells. Overexpression of ERp57 also augmented tunicamycin-induced caspase-3 activation and reduced BiP/GRP78 induction. Our results demonstrate that ERp57 can regulate apoptosis in human endothelial cells. It appears that knockdown of ERp57 confers cellular protection against hyperoxia- or tunicamycin-induced apoptosis by inhibition of caspase-3 activation and stimulation of BiP/GRP78 induction.
Sonic Hedgehog Induces Notch Target Gene Expression in Vascular Smooth Muscle Cells via VEGF-A
Authors: Morrow David , Cullen John P. , Liu Weimin , Guha Shaunta , Sweeney Catherine , Birney Yvonne A. , Collins Nora , Walls Dermot , Redmond Eileen M. , Cahill Paul A.
Published: May 2009

Objective—

Notch, VEGF, and components of the Hedgehog (Hh) signaling pathway have been implicated in vascular morphogenesis. The role of Notch in mediating hedgehog control of adult vascular smooth muscle cell (SMC) growth and survival remains unexplored.

Methods and Results—

In cultured SMCs, activation of Hh signaling with recombinant rShh (3.5 μg/mL) or plasmid encoded Shh increased Ptc1 expression, enhanced SMC growth and survival and promoted Hairy-related transcription factor (Hrt) expression while concomitantly increasing VEGF-A levels. These effects were significantly reversed after Hh inhibition with cyclopamine. Shh-induced stimulation of
Hrt-3
mRNA and SMC growth and survival was attenuated after inhibition of Notch-mediated CBF-1/RBP-Jk-dependent signaling with RPMS-1 while siRNA knockdown of
Hrt-3
inhibited SMC growth and survival. Recombinant VEGF-A increased
Hrt-3
mRNA levels while siRNA knockdown abolished rShh stimulated VEGF-A expression while concomitantly inhibiting Shh-induced increases in
Hrt-3
mRNA levels, proliferating cell nuclear antigen (PCNA), and Notch 1 IC expression, respectively. Hedgehog components were expressed within intimal SMCs of murine carotid arteries after vascular injury concomitant with a significant increase in mRNA for Ptc1, Gli
2
, VEGF-A, Notch 1, and Hrts.

Conclusion—

Hedgehog promotes a coordinate regulation of Notch target genes in adult SMCs via VEGF-A.
Inactivation of the Adenosine A
2A
Receptor Protects Apolipoprotein E–Deficient Mice From Atherosclerosis
Authors: Wang Huan , Zhang Weiyu , Zhu Chuhong , Bucher Christoph , Blazar Bruce R. , Zhang Chunxiang , Chen Jiang-Fan , Linden Joel , Wu Chaodong , Huo Yuqing
Published: May 2009

Background—

Atherosclerosis is a chronic inflammatory disease of the arterial vessel wall. The A
2A
receptor (A
2A
R) plays a central role in many antiinflammatory effects of adenosine. However, the role of A
2A
R in atherosclerosis is not clear.

Methods and Results—

The knockout of A
2A
R in apolipoprotein E–deficient (Apoe
−/−
/A
2A
R
−/−
) mice led to an increase in body weight and levels of blood cholesterol and proinflammatory cytokines, as well as the inflammation status of atherosclerotic lesions. Unexpectedly, Apoe
−/−
/A
2A
R
−/−
mice developed smaller lesions, as did chimeric Apoe
−/−
mice lacking A
2A
R in bone marrow–derived cells (BMDCs). The lesions of those mice exhibited a low density of foam cells and the homing ability of A
2A
R-deficient monocytes did not change. Increased foam cell apoptosis was detected in atherosclerotic lesions of Apoe
−/−
/A
2A
R
−/−
mice. In the absence of A
2A
R, macrophages incubated with oxidized LDL or in vivo–formed foam cells also exhibited increased apoptosis. A
2A
R deficiency in foam cells resulted in an increase in p38 mitogen–activated protein kinase (MAPK) activity. Inhibition of p38 phosphorylation abrogated the increased apoptosis of A
2A
R-deficient foam cells.

Conclusion—

Inactivation of A
2A
R, especially in BMDCs, inhibits the formation of atherosclerotic leisons, suggesting that A
2A
R inactivation may be useful for the treatment of atherosclerosis.
Regulation of Oxygen Distribution in Tissues by Endothelial Nitric Oxide
Authors: Victor Victor M. , Nuñez Cristina , D'Ocón Pilar , Taylor Cormac T. , Esplugues Juan V. , Moncada Salvador
Published: May 2009

Nitric oxide (NO) decreases cellular oxygen (O
2
) consumption by competitively inhibiting cytochrome
c
oxidase. Here, we show that endogenously released endothelial NO, either basal or stimulated, can modulate O
2
consumption both throughout the thickness of conductance vessels and in the microcirculation. Furthermore, we have shown that such modulation regulates O
2
distribution to the surrounding tissues. We have demonstrated these effects by measuring O
2
consumption in blood vessels in a hypoxic chamber and O
2
distribution in the microcirculation using the fluorescent oxygen-probe Ru(phen)
3
2+
. Removal of NO by physical or pharmacological means, or in eNOS
−/−
mice, abolishes this regulatory mechanism. Our results indicate that, in addition to its well-known effect on the regulation of vascular tone, endothelial NO plays a major role in facilitating the distribution of O
2
, an action which is crucial for the adaptation of tissues, including the vessel wall itself, to hypoxia. It is possible that changes in the distribution of O
2
throughout the vessel wall may be implicated in the origin of vascular pathologies such as atherosclerosis.
A Reduced Abbreviated Indirect Calorimetry Protocol Is Clinically Acceptable for Use in Spontaneously Breathing Patients With Traumatic Brain Injury
Authors: McEvoy Claire , Cooke Stephen R. , Young Ian S.
Published: May 2009

A steady state should be established when using indirect calorimetry to assess resting energy expenditure, reduce error, and improve the validity of the measurement. Steady‐state criteria are often rigorous and difficult to achieve in hospitalized patients, particularly in patients recovering from traumatic brain injury. This study aimed to assess whether reducing the timeframe for steady‐state conditions to 2, 3, or 4 minutes when using indirect calorimetry was significantly different from the widely accepted 5‐minute steady‐state protocol in patients recovering from traumatic brain injury. Indirect calorimetry using an open‐circuit calorimeter was used to measure resting energy expenditure in 20 spontaneously breathing patients with traumatic brain injury. Each patient underwent repeated measurements once per day at various stages of his or her recovery. Bland‐Altman methods comparison was used for statistical analysis. In total, 124 measurements were recorded, of which 59% met 5‐minute, 70% met 4‐minute, 76% met 3‐minute, and 84% met 2‐minute steady‐state conditions. Mean within‐patient coefficient of variation was 13% (range, 0%‐35%). Agreement at the ±10% level using Bland‐Altman plots was 100% for a 4‐minute steady‐state protocol, 96% for a 3‐minute protocol, and 90% using a 2‐minute protocol. A 4‐minute or 3‐minute abbreviated steady‐state indirect calorimetry protocol is clinically acceptable to measure resting energy expenditure in spontaneously breathing patients with a traumatic brain injury.
Analysis of the complement and molecular evolution of tRNA genes in cow
Authors: Tang Dave TP , Glazov Evgeny A , McWilliam Sean M , Barris Wesley C , Dalrymple Brian P
Published: April 2009

Background

Detailed information regarding the number and organization of transfer RNA (tRNA) genes at the genome level is becoming readily available with the increase of DNA sequencing of whole genomes. However the identification of functional tRNA genes is challenging for species that have large numbers of repetitive elements containing tRNA derived sequences, such as Bos taurus. Reliable identification and annotation of entire sets of tRNA genes allows the evolution of tRNA genes to be understood on a genomic scale.

Results

In this study, we explored the B. taurus genome using bioinformatics and comparative genomics approaches to catalogue and analyze cow tRNA genes. The initial analysis of the cow genome using tRNAscan-SE identified 31,868 putative tRNA genes and 189,183 pseudogenes, where 28,830 of the 31,868 predicted tRNA genes were classified as repetitive elements by the RepeatMasker program. We then used comparative genomics to further discriminate between functional tRNA genes and tRNA-derived sequences for the remaining set of 3,038 putative tRNA genes. For our analysis, we used the human, chimpanzee, mouse, rat, horse, dog, chicken and fugu genomes to predict that the number of active tRNA genes in cow lies in the vicinity of 439. Of this set, 150 tRNA genes were 100% identical in their sequences across all nine vertebrate genomes studied. Using clustering analyses, we identified a new tRNA-GlyCCC subfamily present in all analyzed mammalian genomes. We suggest that this subfamily originated from an ancestral tRNA-GlyGCC gene via a point mutation prior to the radiation of the mammalian lineages. Lastly, in a separate analysis we created phylogenetic profiles for each putative cow tRNA gene using a representative set of genomes to gain an overview of common evolutionary histories of tRNA genes.

Conclusion

The use of a combination of bioinformatics and comparative genomics approaches has allowed the confident identification of a set of cow tRNA genes that will facilitate further studies in understanding the molecular evolution of cow tRNA genes.
Lack of association between polymorphisms of MASP2and susceptibility to SARS coronavirus infection
Authors: Wang Yan , Yan Jiangwei , Shi Yuling , Li Ping , Liu Chuanxuan , Ma Qingjun , Yang Ruifu , Wang Xiaoyi , zhu Lina , Yang Xiao , Cao Cheng
Published: May 2009

Background

The pathogenesis of severe acute respiratory disease syndrome (SARS) is not fully understood. One case-control study has reported an association between susceptibility to SARS andmannan-binding lectin(MBL) in China. As the downstream protein ofMBL, variants of theMBL-associated serine protease-2 (MASP2) gene may be associated with SARS coronavirus (SARS-CoV) infection in the same population.

Methods

Thirty individuals with SARS were chosen for analysis ofMASP2polymorphisms by means of PCR direct sequencing. Tag single nucleotide polymorphisms (tagSNPs) were chosen using pairwise tagging algorithms. The frequencies of four tag SNPs (rs12711521, rs2261695, rs2273346 and rs7548659) were ascertained in 376 SARS patients and 523 control subjects, using the Beckman SNPstream Ultra High Throughput genotyping platform.

Results

There is no significant association between alleles or genotypes of theMASP2tagSNP and susceptibility to SARS-CoV in both Beijing and Guangzhou populations. Diplotype (rs2273346 and rs12711521)were analyzed for association with susceptibility to SARS, no statistically significant evidence of association was observed. The Beijing and Guangzhou sample groups were homogeneous regarding demographic and genetic parameters, a joined analysis also showed no statistically significant evidence of association.

Conclusion

Our data do not suggest a role forMASP2polymorphisms in SARS susceptibility in northern and southern China.