bluuebunny/crossref_metadata_2025 huggingface dataset:
Human bone marrow stromal cells simultaneously support B and T/NK lineage development from human haematopoietic progenitors: a principal role for flt3 ligand in lymphopoiesis
Authors: Nakamori Yoshiki , Liu Bing , Ohishi Kohshi , Suzuki Kei , Ino Kazuko , Matsumoto Takeshi , Masuya Masahiro , Nishikawa Hiroyoshi , Shiku Hiroshi , Hamada Hirofumi , Katayama Naoyuki
Published: April 2012

The regulation of human early lymphopoiesis remains unclear. B‐ and T‐lineage cells cannot develop simultaneously with conventional stromal cultures. Here we show that telomerized human bone marrow stromal cells supported simultaneous generation of CD19+CD34lo/−CD10+cyCD79a+CD20+/−VpreB− pro‐B cells and CD7+CD34+ CD45RA+CD56−cyCD3− early T/Natural Killer (NK) cell precursors from human haematopoietic progenitors, and the generation of both lymphoid precursors was promoted by flt3 ligand (flt3L). On the other hand, stem cell factor or thrombopoietin had little or no effect when used alone. However, both acted synergistically with flt3L to augment the generation of both lymphoid precursors. Characteristics of these lymphoid precursors were evaluated by gene expression profiles, rearrangements of IgH genes, or replating assays. Similar findings were observed with primary human bone marrow stromal cells. Notably, these two lymphoid‐lineage precursors were generated without direct contact with stromal cells, indicating that early B and T/NK development can occur, at least in part, by stromal cell‐derived humoral factors. In serum‐free cultures, flt3L elicited similar effects and appeared particularly important for B cell development. The findings of this study identified the potential of human bone marrow stromal cells to support human early B and T lymphopoiesis and a principal role for flt3L during early lymphopoiesis.
‘I am part of every decision’: client perceptions of engagement within a supportive housing child welfare programme
Authors: Farrell Anne F. , Luján Melissa L. , Britner Preston A. , Randall Kellie G. , Goodrich Samantha A.
Published: April 2012

The majority of children in the child welfare system enter because of neglect and come from poor families with high rent burden, substandard housing and risk for homelessness. In this paper, we describe a model programme for families with dual vulnerability in housing and child welfare. Clients presented with a variety of parenting, substance use and/or mental health issues. The Supportive Housing for Families (SHF) programme prioritizes prompt family access to housing and related supports and operates from an intensive, family‐centred casework that promotes client engagement as a mechanism for change. We used a mixed methods approach that included the administration of Alpert and Britner's Parent Engagement Measure (quantitative) and open‐ended interviews (qualitative) with 41 parents involved in the child welfare system. Results indicate high levels of client engagement, with convergence across the formal measure and interview themes. SHF promoted client engagement through the swift provision of tangible resources, as well as caseworker resourcefulness and responsiveness. The Parent Engagement Measure performed well psychometrically. We compare findings with prior research and discuss implications, limitations and future directions.
Assessment: when does it help and when does it hinder? Parents' experiences of the assessment process
Authors: Harris Nathan
Published: April 2012

It is widely acknowledged that there are significant benefits when statutory child protection agencies and parents are able to engage cooperatively with one another to ensure the well‐being of children. Despite promising innovations there are ongoing concerns that the current child protection model alienates and confuses many parents. It is hypothesized that a significant reason for this is that statutory agencies have become reliant on formalistic assessment, and as a consequence interactions with parents have become dominated by a focus on assessment‐compliance. A qualitative study of 40 parents who had recently been investigated in Australia is reported. The analysis suggests that many parents find characteristics of assessment processes intrusive and that this undermines engagement. It is concluded that there should be greater debate about the role that assessment plays in child protection practice.
A secure base? The adolescent–staff relationship in secure residential youth care
Authors: Harder Annemiek T. , Knorth Erik J. , Kalverboer Margrite E.
Published: April 2012

The client–therapist relationship is recognized as an important factor contributing to outcomes of child and youth care. Particularly in secure residential youth care, in which adolescents with mainly externalizing behaviour problems are often placed coercively, the client–staff relationship seems to be important for the achievement of positive outcomes. The present study aims to assess the client–staff relationship for a group of 135 adolescents in secure residential care and factors associated with a positive relationship. The results show that adolescents, group care workers and teachers experience a limited affective bond in their relationship 2 months after the adolescents were admitted to the secure care centre. Adolescents do tend to use care workers and teachers as a secure attachment figure, which suggests that an affective bond is no precondition for the adolescents to experience staff as a secure base. Main predictors of a good relationship are the positive treatment skills of both group care workers and teachers. These findings point to the need for training of care workers and teachers so that they are better prepared for working with these adolescents.
Dehydroxymethylepoxyquinomicin (DHMEQ), a novel NF‐kappaB inhibitor, inhibits allergic inflammation and airway remodelling in murine models of asthma
Authors: Shimizu K. , Konno S. , Ozaki M. , Umezawa K. , Yamashita K. , Todo S. , Nishimura M.
Published: April 2012

Background

Dehydroxymethylepoxyquinomicin (DHMEQ) is a newly developed compound that inhibits nuclear factor κB activation and is reported to ameliorate animal models of various inflammatory diseases without significant adverse effects. Because nuclear factor κB is a transcription factor that plays a critical role in the pathophysiology of asthma, DHMEQ may be of therapeutic benefit in asthma.

Objective

The purpose of this study was to evaluate the effects of DHMEQ on airway inflammation and remodelling in murine models of asthma.

Methods

The BALB/c mice were sensitized and then challenged acutely or chronically with ovalbumin and administered DHMEQ intraperitoneally before each challenge. Inflammation of airways, lung histopathology and airway hyper responsiveness to methacholine challenge were evaluated. In addition, the effect of DHMEQ on production of cytokines and eotaxin‐1 by murine splenocytes, human peripheral blood mononuclear cells and bronchial epithelial cells was investigated.

Results

Airway hyper responsiveness was ameliorated in both acutely and chronically challenged models by treatment with DHMEQ. DHMEQ significantly reduced eosinophilic airway inflammation and levels of Th2 cytokines in bronchoalveolar lavage fluid in the acute model. It also inhibited parameters of airway remodelling including mucus production, peribronchial fibrosis and the expression of α‐smooth muscle actin. Moreover, the production of Th2 cytokines from murine splenocytes and human peripheral blood mononuclear cells and the production of eotaxin‐1 by bronchial epithelial cells were inhibited by DHMEQ.

Conclusions and Clinical Relevance

These results indicate that DHMEQ inhibits allergic airway inflammation and airway remodelling in murine models of asthma. DHMEQ may have therapeutic potential in the treatment of asthma.
Phenotyping airways disease: an A to E approach
Authors: Gonem S. , Raj V. , Wardlaw A. J. , Pavord I. D. , Green R. , Siddiqui S.
Published: April 2012

The airway diseases asthma and chronic obstructive pulmonary disease (COPD) are heterogeneous conditions with overlapping pathophysiological and clinical features. It has previously been proposed that this heterogeneity may be characterized in terms of five relatively independent domains labelled from A to E, namely airway hyperresponsiveness (AHR), bronchitis, cough reflex hypersensitivity, damage to the airways and surrounding lung parenchyma, and extrapulmonary factors. Airway hyperresponsiveness occurs in both asthma and COPD, accounting for variable day to day symptoms, although the mechanisms most likely differ between the two conditions. Bronchitis, or airway inflammation, may be predominantly eosinophilic or neutrophilic, with different treatments required for each. Cough reflex hypersensitivity is thought to underlie the chronic dry cough out of proportion to other symptoms that can occur in association with airways disease. Structural changes associated with airway disease (damage) include bronchial wall thickening, airway smooth muscle hypertrophy, bronchiectasis and emphysema. Finally, a variety of extrapulmonary factors may impact upon airway disease, including rhinosinusitis, gastroesophageal reflux disease, obesity and dysfunctional breathing. This article discusses the A to E concept in detail and describes how this framework may be used to assess and treat patients with airway diseases in the clinic.
Effects of natural S‐equol supplements on overweight or obesity and metabolic syndrome in the Japanese, based on sex and equol status
Authors: Usui Takeshi , Tochiya Mayu , Sasaki Yousuke , Muranaka Kazuya , Yamakage Hajime , Himeno Akihiro , Shimatsu Akira , Inaguma Asami , Ueno Tomomi , Uchiyama Shigeto , Satoh‐Asahara Noriko
Published: April 2012

Objectives

Epidemiologic studies indicate that soy intake has an important role in the prevention of age‐related health problems. Daidzein, the principal isoflavone contained in soy, is converted to S‐equol by the intestinal bacteria. Not all individuals, however, can produce S‐equol, which is considered the most biologically active metabolite. We studied the effects of a natural S‐equol supplement on metabolic parameters associated with overweight or obesity and metabolic syndrome.

Methods

The study was a randomized, double‐blinded, placebo‐controlled, crossover design with no washout period. All subjects were considered overweight or obese if they had a body mass index ≧25 kg/m2. Placebo or natural S‐equol tablets containing 10 mg S‐equol were orally ingested each day for 12 weeks. A total of 54 Japanese overweight or obese outpatients were enrolled. The equol phenotype was determined, and various metabolic parameters, including cardio‐ankle vascular index (CAVI), were measured.

Results

Equol non‐producers comprised 67·9% of the overweight or obese subjects. The ratio of equol non‐producers in this overweight or obese subject group was higher than the previously reported ratio of equol non‐producers (approximately 50%) in the general population. Compared with the placebo group, intervention with natural S‐equol led to a significant decrease in HbA1c, serum low‐density lipoprotein cholesterol (LDL‐C) levels and CAVI score. Furthermore, the effect was more prominent in the subgroup of female equol non‐producers.

Conclusion

The ratio of equol non‐producers in overweight or obese populations might be higher than generally reported. Natural S‐equol might have a role in glycaemic control and in the prevention of cardiovascular disease by its effects to lower LDL‐C levels and CAVI scores in overweight or obese individuals.
Associations between visceral adipose tissue, inflammation and sex steroid concentrations in men
Authors: Gautier Alain , Bonnet Fabrice , Dubois Séverine , Massart Catherine , Grosheny Catherine , Bachelot Anne , Aubé Christophe , Balkau Beverley , Ducluzeau Pierre‐Henri
Published: April 2012

Context

In men, obesity and the metabolic syndrome are accompanied by decreased testosterone levels, but little is known about the associations between visceral adipose tissue (VAT), VAT‐related inflammation and sex steroids.

Objective

To examine the relative impact of VAT, abdominal subcutaneous adipose tissue (SAT) and interleukin 6 (IL‐6), a marker of VAT‐induced inflammation, on testosterone (T) and 17β‐oestradiol (E2) levels in dysmetabolic men.

Methods

We study the NUMEVOX cohort of 229 men, aged 27–77 years, who all had at least one metabolic syndrome criterion (on average three). IL‐6, C‐reactive protein, Homeostasis Model Assessment of (HOMA) insulin resistance index (HOMA‐IR), liver enzymes, E2, LH, sex hormone‐binding globulin (SHBG), T, waist circumference and body mass index (BMI) were measured; bioavailable testosterone (BT) was calculated from T and SHBG; MRI‐assessed VAT and SAT were analysed in 109 of these men.

Results

Visceral adipose tissue was strongly correlated with E2 (Spearman r = 0·38, P < 0·001) and with BT/E2 ratio (r = −0·42, P < 0·001), while SAT was not correlated with either. IL‐6 was correlated with E2 (r = 0·19, P = 0·007), BT (r = −0·19, P = 0·006) and BT/E2 ratio (r = −0·30 P < 0·001). In multivariate linear analysis, the relation between VAT and E2 was independent of age, BMI (P = 0·008), leptin (P < 0·001), T and SHBG. Log(IL‐6) was significantly inversely related with log(BT) (P = 0·032) independently of age, VAT, leptin and HOMA‐IR.

Conclusions

17β‐oestradiol levels were positively associated with VAT, but not with SAT, while T and BT were negatively and independently associated with IL‐6. The significant inverse association between IL‐6 and T suggests an important role of low‐grade visceral fat inflammation in the central hypogonadism associated with the metabolic syndrome.
Healthy birth after testicular extraction of sperm and ICSI from an azoospermic man with mild androgen insensitivity syndrome caused by an androgen receptor partial loss‐of‐function mutation
Authors: Massin Nathalie , Bry Hélène , Vija Lavinia , Maione Luigi , Constancis Elizabeth , Haddad Bassam , Morel Yves , Claessens Frank , Young Jacques
Published: April 2012

Context

The androgen receptor (AR) is essential for the development and maintenance of the male phenotype, and for spermatogenesis. Mutations in the AR gene cause a wide variety of androgen insensitivity syndromes (AIS), ranging from complete feminization to phenotypic males with infertility.

Objective

We report the first birth achieved after intracytoplasmic sperm injection (ICSI) with sperm from an azoospermic man with an AR mutation associated with mild AIS (MAIS).

Patients and methods

A couple with primary infertility was referred to our centre. The man had azoospermia with testicular hypotrophy and an undervirilized phenotype despite a normal plasma testosterone level. His androgen sensitivity index and serum anti‐mullerian hormone (AMH) levels were elevated, pointing to AIS. Molecular analysis of the AR gene revealed a point mutation resulting in an F754S substitution (renumbered F755S in the 2012 McGill University AR gene database), in the ligand‐binding domain of the protein, and further analysis indicated impaired receptor function.

Results

After genetic counselling of the couple, oocytes were retrieved after controlled ovarian hyperstimulation, and sperm were obtained simultaneously by testicular extraction for ICSI. Nine embryos were obtained. Two were transferred and two were suitable for cryopreservation. A pregnancy was obtained and a healthy girl, carrying the F754S AR mutation, was born at 37 weeks of gestation. AR and AMH were detected by immunohistochemistry in the patient's testicular specimens. AMH immuno‐staining was intense in tubules without spermatogenesis and weak in those with ongoing spermatogenesis.

Conclusion

A healthy child can be obtained by testicular extraction and ICSI despite azoospermia in MAIS. The parents must be informed of the X‐linked transmission of the mutation to their descendants. The relationship between AR signalling, testicular AMH expression and spermatogenesis in this patient is discussed.
What are the keys to successful thyroid FNA interpretation?
Authors: Sebo Thomas J.
Published: April 2012

There is much concern expressed in the literature regarding the lack of predictive power of the thyroid fine needle aspiration/biopsy (TFNAB) approach to defining the nature of clinically detected thyroid nodules. This has been exacerbated in the past decade or more by the routine use of ultrasonography (US) in examining the thyroid as well as the introduction of molecular testing in the realm of thyroid pathology. Some have even gone so far as to suggest replacing the TFNAB with molecular signature testing to reduce the degree of uncertainty for a specific cytological diagnosis. This review addresses those concerns with a re‐emphasis on understanding the basic keys to successfully evaluating a patient with a thyroid nodule by routine TFNAB examination. These keys include bringing to bear an experienced group of physicians in an integrated team approach, a sharpened focus on the TFNAB cytological categories and a grounded understanding of the predictive ability of molecular testing in a given patient when the cytological interpretation creates too much uncertainty in the minds of the patients and clinicians in attempting to reach a decision on how to manage a thyroid nodule. With this practical approach in mind, the false‐negative and false‐positive rates of “negative (benign)” and “positive (malignant)” thyroid aspirates should be no more than 1%; and the prevalence of an “indeterminate” aspirate – the area in TFNAB attracting the most attention for improvement with novel biomarkers – should be 10% or less. Thus, physicians should be capable of managing at least 90% of patients undergoing TFNAB in a confident manner without further testing beyond the routine, future re‐examination of the patient's nodule to re‐assess for any change in its nature or its impact on the patient's quality of life. The other 10% can then be considered for molecular testing in a manner tailored to those individuals truly in need of a more sophisticated – and expensive – approach to the characterization of their thyroid nodules. As the era of US has matured – just as with the era of serum PSA testing for prostate cancer in men – we have experienced a paradigm shift: Given the incidence of thyroid nodules in the U.S. adult population in comparison with the risk of dying of thyroid malignancy, the main role of TFNAB is to reduce the need for surgical intervention.
Trophic positioning and microphytobenthic carbon uptake of biofilm‐dwelling meiofauna in a temperate river
Authors: MAJDI NABIL , TACKX MICHÈLE , BUFFAN–DUBAU EVELYNE
Published: April 2012

1. δ13C and δ15N stable isotope signatures combined with an in situ microphytobenthic 13C labelling experiment were performed on epilithic biofilms of a large temperate river (the Garonne, France) to infer the trophic positioning of biofilm‐dwelling meiofauna and their uptake of microphytobenthic carbon.

2. Chironomidae larvae and Chromadorina spp. nematodes rapidly incorporated freshly produced microphytobenthic carbon in contrast to Rhyacophilidae larvae and Naididae oligochaetes. Quantitatively, macrofaunal Chironomidae incorporated more microphytobenthic carbon per day than did meiofauna. Moreover, Chironomidae seemed more involved in the spatial export of microphytobenthic carbon than nematodes.

3. Rhyacophilidae larvae were predators feeding on large meiofauna (Naididae and Chironomidae) but not on nematodes. Naididae oligochaetes primarily gained their carbon from allochthonous and/or microbial‐loop recycled sources.

4. A rapid and significant loss of labelled microphytobenthic carbon was observed. Feeding activity of biofilm‐dwelling invertebrates seemed not to be primarily involved in this loss.
No significant nitrous oxide emissions during spring thaw under grazing and nitrogen addition in an alpine grassland
Authors: Li Kaihui , Gong Yanming , Song Wei , Lv Jinling , Chang Yunhua , Hu Yukun , Tian Changyan , Christie Peter , Liu Xuejun
Published: April 2012

A recent study (Wolf et al., 2010) suggests that short—lived pulses of N2O emission during spring thaw dominate the annual N2O budget and that grazing decreases N2O emissions during the spring thaw. To verify this we conducted year—round N2O flux measurements from June 2010 to May 2011 in Tianshan alpine grassland in central Asia. No pulse emissions of N2O were found at grazing management sites and nitrogen addition sites during the spring thaw. The contribution of the spring thaw to the total annual N2O budget was small and accounted for only 6.6% of the annual fluxes, with winter emissions accounting for 16.7% and growing season emissions accounting for 76.7%. The difference in N2O emissions attributable to grazing management was not significant (P > 0.05). Nitrogen input tended to increase N2O emissions at N addition sites during the grass growing season compared with those at unfertilized sites. N2O fluxes showed a significant correlation with air temperature and also with both soil temperature and soil water content at 10 cm depth.
Conserving energy at a cost to biodiversity? Impacts of LED lighting on bats
Authors: Stone Emma L. , Jones Gareth , Harris Stephen
Published: April 2012

Artificial lighting is a key biodiversity threat and produces 1900 million tonnes of
CO

2 emissions globally, more than three times that produced by aviation. The need to meet climate change targets has led to a global increase in energy‐efficient light sources such as high‐brightness light‐emitting diodes (LEDs). Despite the energetic benefits of LEDs, their ecological impacts have not been tested. Using an experimental approach, we show that LED street lights caused a reduction in activity of slow‐flying bats (
Rhinolophus hipposideros and
Myotis spp.). Both
R. hipposideros and
Myotis spp. activities were significantly reduced even during low light levels of 3.6 lux. There was no effect of LED lighting on the relatively fast‐flying
Pipistrellus pipistrellus, Pipistrellus pygmaeus and
Nyctalus/Eptesicus spp. We provide the first evidence of the effects of LED lights on bats. Despite having considerable energy‐saving benefits, LED lights can potentially fragment commuting routes for bats with associated negative conservation consequences. Our results add to the growing evidence of negative impacts of lighting on a wide range of taxa. We highlight the complexities involved in simultaneously meeting targets for reduction of greenhouse gas emissions and biodiversity loss. New lighting strategies should integrate climate change targets with the cultural, social and ecological impacts of emerging lighting technologies.
Increased temperature and altered summer precipitation have differential effects on biological soil crusts in a dryland ecosystem
Authors: Johnson Shannon L. , Kuske Cheryl R. , Carney Travis D. , Housman David C. , Gallegos‐Graves La Verne , Belnap Jayne
Published: April 2012

Biological soil crusts (biocrusts) are common and ecologically important members of dryland ecosystems worldwide, where they stabilize soil surfaces and contribute newly fixed C and N to soils. To test the impacts of predicted climate change scenarios on biocrusts in a dryland ecosystem, the effects of a 2–3 °C increase in soil temperature and an increased frequency of smaller summer precipitation events were examined in a large, replicated field study conducted in the cold desert of the Colorado Plateau, USA. Surface soil biomass (DNA concentration), photosynthetically active cyanobacterial biomass (chlorophyll a concentration), cyanobacterial abundance (quantitative PCR assay), and bacterial community composition (16S rRNA gene sequencing) were monitored seasonally over 2 years. Soil microbial biomass and bacterial community composition were highly stratified between the 0–2 cm depth biocrusts and 5–10 cm depth soil beneath the biocrusts. The increase in temperature did not have a detectable effect on any of the measured parameters over 2 years. However, after the second summer of altered summer precipitation pattern, significant declines occurred in the surface soil biomass (avg. DNA concentration declined 38%), photosynthetic cyanobacterial biomass (avg. chlorophyll a concentration declined 78%), cyanobacterial abundance (avg. gene copies g−1 soil declined 95%), and proportion of Cyanobacteria in the biocrust bacterial community (avg. representation in sequence libraries declined 85%). Biocrusts are important contributors to soil stability, soil C and N stores, and plant performance, and the loss or reduction of biocrusts under an altered precipitation pattern associated with climate change could contribute significantly to lower soil fertility and increased erosion and dust production in dryland ecosystems at a regional scale.
Forage production and nutritive value of Dactylis glomerata and Trifolium subterraneum mixtures under different shading treatments
Authors: Kyriazopoulos A. P. , Abraham E. M. , Parissi Z. M. , Koukoura Z. , Nastis A. S.
Published: April 2012

Production and nutritive value of forage in silvopastural systems can be improved by introducing shade‐ tolerant grass and legume species in appropriate mixtures. The management of these systems can present a challenge regarding the selection of the proper grass and legume species as well as the maintenance of the optimum balance between the two species in the grass–legume stand. The objectives of the present study were to evaluate the performance of pure stands and mixtures (75:25, 50:50, 25:75) of Dactylis glomerata:Trifolium subterraneum under full sun, 60% shade and 90% shade. Dry matter production of D. glomerata was not affected by shading, while that of T. subterraneum was drastically reduced. Shading increased the crude protein (CP) and acid detergent lignin content, but did not affect the acid detergent fibre and neutral detergent fibre content. The dry matter production of the 75:25 and 25:75 mixtures was higher compared to the one of the 50:50 mixture, while the CP content of the 75:25 mixture was the highest under moderate shading. However, relative yield, aggressivity index and competitive balance index indicated higher competitive ability for D. glomerata compared with T. subterraneum especially under shade. Therefore, the 25:75 mixture is suggested as the most suitable under moderate shade (60%), to perpetuate the stand and avoid the frequent re‐sowing of T. subterraneum.
Chemical–physical properties of TheraCal, a novel light‐curable MTA‐like material for pulp capping
Authors: Gandolfi M. G. , Siboni F. , Prati C.
Published: April 2012

Gandolfi MG, Siboni F, Prati C. Chemical–physical properties of TheraCal, a novel light‐curable MTA‐like material for pulp capping. International Endodontic Journal, 45, 571–579, 2012.

Aim  To evaluate the chemical–physical properties of TheraCal, a new light‐curable pulp‐capping material composed of resin and calcium silicate (Portland cement), compared with reference pulp‐capping materials (ProRoot MTA and Dycal).

Methodology  Calcium (Ca) and hydroxyl (OH) ion release over 28 days, solubility and water uptake (weight percentage variation, Δ%) at 24 h, cure depth and radiopacity of TheraCal, ProRoot MTA and Dycal were evaluated. Statistical analysis (P < 0.05) of release of ion was carried out by two‐way repeated measures anova with Tukey, whilst one‐way anova with Tukey test was used for the other tests.

Results  TheraCal released significantly more calcium than ProRoot MTA and Dycal throughout the test period. TheraCal was able to alkalinize the surrounding fluid initially to pH 10–11 (3 h–3 days) and subsequently to pH 8–8.5 (7–14 days). TheraCal had a cure depth of 1.7 mm. The solubility of TheraCal (Δ−1.58%) was low and significantly less than that of Dycal (Δ−4.58%) and ProRoot MTA (Δ−18.34%). The amount of water absorbed by TheraCal (Δ +10.42%) was significantly higher than Dycal (Δ +4.87%) and significantly lower than ProRoot MTA (Δ +13.96%).

Conclusions   TheraCal displayed higher calcium‐releasing ability and lower solubility than either ProRoot MTA or Dycal. The capability of TheraCal to be cured to a depth of 1.7 mm may avoid the risk of untimely dissolution. These properties offer major advantages in direct pulp‐capping treatments.
T‐cell phenotypes in bronchoalveolar lavage fluid, blood and lymph nodes in pulmonary sarcoidosis – indication for an airborne antigen as the triggering factor in sarcoidosis
Authors: Darlington P. , Haugom‐Olsen H. , von Sivers K. , Wahlström J. , Runold M. , Svjatoha V. , Porwit A. , Eklund A. , Grunewald J.
Published: April 2012

Abstract.  Darlington P, Haugom‐Olsen H, von Sivers K, Wahlström J, Runold M, Svjatoha V, Porwit A, Eklund A, Grunewald J (Karolinska Institutet, Stockholm, Sweden). T‐cell phenotypes in bronchoalveolar lavage fluid, blood and lymph nodes in pulmonary sarcoidosis – indication for an airborne antigen as the triggering factor in sarcoidosis. J Intern Med 2012; 272: 465–471.

Background.  An increased percentage of CD4+ T cells is usually observed in bronchoalveolar lavage fluid (BALF) from patients with sarcoidosis. In HLA‐DRB1*03‐positive patients, such T cells express the T‐cell receptor (TCR) AV2S3+ gene segment. It is not known whether cells found in BALF reflect those in enlarged regional lymph nodes (LNs). Therefore, the aim of this study was to compare T‐cell phenotypes in BALF, blood and mediastinal LNs.

Methods.  Fifteen patients underwent clinical investigation including bronchoscopy with bronchoalveolar lavage. Blood samples were drawn, and endoscopic ultrasound‐guided fine‐needle aspiration of enlarged mediastinal LNs was performed via the oesophagus. T cells from all three compartments were analysed by flow cytometry for markers of activity, differentiation and T regulatory function.

Results.  The CD4/CD8 ratio was significantly higher in BALF compared with regional LNs and was also significantly higher in LNs than in blood. The CD4+ T cells were recently activated and more differentiated in BALF than in blood and LNs. There was an accumulation of T regulatory cells (FOXP3+) in LNs and a correlation between high levels of FOXP3+ cells in BALF and in LNs. In HLA‐DRB1*03‐positive patients, TCR AV2S3+ CD4+ T cells were predominantly localized within BALF.

Conclusions.  The CD4+ T‐cell phenotype in BALF indicates an active ongoing specific immune response primarily localized to the alveolar space.
The Bacillus subtilis cannibalism toxin SDP collapses the proton motive force and induces autolysis
Authors: Lamsa Anne , Liu Wei‐Ting , Dorrestein Pieter C. , Pogliano Kit
Published: April 2012

Bacillus subtilis SDP is a peptide toxin that kills cells outside the biofilm to support continued growth. We show that purified SDP acts like endogenously produced SDP; it delays sporulation, and the SdpI immunity protein confers SDP resistance. SDP kills a variety of Gram‐positive bacteria in the phylum Firmicutes, as well as Escherichia coli with a compromised outer membrane, suggesting it participates in defence of the B. subtilis biofilm against Gram‐positive bacteria as well as cannibalism. Fluorescence microscopy reveals that the effect of SDP on cells differs from that of nisin, nigericin, valinomycin and vancomycin‐KCl, but resembles that of CCCP, DNP and azide. Indeed, SDP rapidly collapses the PMF as measured by fluorometry and flow cytometry, which triggers the slower process of autolysis. This secondary consequence of SDP treatment is not required for cell death since the autolysin‐defective lytC, lytD, lytE, lytF strain fails to be lysed but is nevertheless killed by SDP. Collapsing the PMF is an ideal mechanism for a toxin involved in cannibalism and biofilm defence, since this would incapacitate neighbouring cells by inhibiting motility and secretion of proteins and toxins. It would also induce autolysis in many Gram‐positive species, thereby releasing nutrients that promote biofilm growth.
Role of a Candida albicans Nrm1/Whi5 homologue in cell cycle gene expression and DNA replication stress response
Authors: Ofir Ayala , Hofmann Kay , Weindling Esther , Gildor Tsvia , Barker Katherine S. , Rogers P. David , Kornitzer Daniel
Published: April 2012

To explore cell cycle regulation in the dimorphic fungus Candida albicans, we identified and characterized CaNrm1, a C. albicans homologue of the Saccharomyces cerevisiae Whi5 and Nrm1 transcription inhibitors that, analogous to mammalian Rb, regulate the cell cycle transcription programme during the G1 phase. CaNRM1 is able to complement the phenotypes of both whi5 and nrm1 mutants in S. cerevisiae. In C. albicans, global transcription analysis of the CaNRM1 deletion mutant reveals a preferential induction of G1‐ and G1/S‐specific genes. CaNrm1 interacts genetically with the C. albicans MBF functional homologue, and physically with its subunit CaSwi4. Similar to S. cerevisiae Whi5, CaNrm1 subcellular localization oscillates with the cell cycle between the nucleus and the cytoplasm. Deletion of CaNRM1 further results in increased resistance to hydroxyurea, an inhibitor of DNA replication; analysis of the expression of ribonucleotide reductase, the target of hydroxyurea, suggests that its transcriptional induction in response to hydroxyurea is regulated via CaNrm1, and biochemical analysis shows that hydroxyurea causes disruption of the interaction of CaNrm1 with CaSwi4. Furthermore, induction of the hyphal‐specific genes is dampened under certain conditions in the Canrm1−/− mutant, suggesting that the cell cycle transcription programme can influence the morphogenetic transcription programme of C. albicans.
Manganese is required for oxidative metabolism in unstressed Bradyrhizobium japonicum cells
Authors: Hohle Thomas H. , O'Brian Mark R.
Published: April 2012

Recent studies of Mn2+ transport mutants indicate that manganese is essential for unstressed growth in some bacterial species, but is required primarily for induced stress responses in others. A Bradyrhizobium japonicum mutant defective in the high‐affinity Mn2+ transporter gene mntH has a severe growth phenotype under manganese limitation, suggesting a requirement for the metal under unstressed growth. Here, we found that activities of superoxide dismutase and the glycolytic enzyme pyruvate kinase were deficient in an mntH strain grown under manganese limitation. We identified pykM as the only pyruvate kinase‐encoding gene based on deficiency in activity of a pykM mutant, rescue of the growth phenotype with pyruvate, and pyruvate kinase activity of purified recombinant PykM. PykM is unusual in that it required Mn2+ rather than Mg2+ for high activity, and that neither fructose‐1,6‐bisphosphate nor AMP was a positive allosteric effector. The mntH‐dependent superoxide dismutase is encoded by sodM, the only expressed superoxide dismutase‐encoding gene under unstressed growth conditions. An mntH mutant grew more slowly on pyruvate under manganese‐limited conditions than did a pykM sodM double mutant, implying additional manganese‐dependent processes. The findings implicate roles for manganese in key steps in unstressed oxidative metabolism in B. japonicum.
IgG Subclasses Against Helicobacter pylori Isolates: An important Tool for Disease Characterization
Authors: Martínez‐Becerra F. , Castillo‐Rojas G. , de León S. Ponce , López‐Vidal* Y.
Published: April 2012

Helicobacter pylori infects around 50% of the world’s population and is associated with diverse pathologies. In the most severe cases, the bacterium causes peptic ulcers and gastric cancer. The interplay between H. pylori and the host’s immune response may help to determine the specific outcome of the infection. To study the relationship between antibody subclasses and variation in immune recognition, we determined the immunoglobulin G1 and 2 (IgG1 and IgG2) titres of sera obtained from patients with different H. pylori‐associated pathologies. IgG1 and IgG2 titres were determined by ELISA in 44 sera of patients with different H. pylori‐associated diseases (peptic ulcer, bleeding peptic ulcers, gastric cancer and dyspepsia). Soluble proteins from lysates were obtained from 12 different clinical isolates from similar associated diseases. We found that soluble proteins from lysates of H. pylori strains (SPLHP) recognition patterns in these sera were highly variable. Overall, IgG2 titres were higher than the IgG1 titres in the infected patients. In particular, those with peptic ulcers showed marked elevation in IgG2/IgG1 ratios, while SPLHPs from dyspeptic patients resulted in high IgG1 titres. Our results reveal that correlation of antibody subclass titres with Th1/Th2 markers may aid pathology characterization and show a potential diagnosis that could be formally evaluated in other studies.
Langerhans Cells and T Cells Sense Cell Dysplasia in Oral Leukoplakias and Oral Squamous Cell Carcinomas – Evidence for Immunosurveillance
Authors: Öhman J. , Magnusson B. , Telemo E. , Jontell M. , Hasséus B.
Published: April 2012

Leukoplakias (LPLs) are lesions in the oral mucosa that may develop into oral squamous cell carcinoma (OSCC). The objective of this study was to assess presence and distribution of dendritic Langerhans cells (LCs) and T cells in patients with LPLs with or without cell dysplasia and in oral squamous cell carcinoma (OSCC). Biopsy specimens from patients with leukoplakias (LPLs) with or without dysplasia and oral squamous cell carcinoma (OSCC) were immunostained with antibodies against CD1a, Langerin, CD3, CD4, CD8 and Ki67, followed by quantitative analysis. Analyses of epithelium and connective tissue revealed a significantly higher number of CD1a + LCs in LPLs with dysplasia compared with LPLs without dysplasia. Presence of Langerin + LCs in epithelium did not differ significantly between LPLs either with or without dysplasia and OSCC. T cells were found in significantly increased numbers in LPLs with dysplasia and OSCC. The number of CD4+ cells did not differ significantly between LPLs with and without dysplasia, but a significant increase was detected when comparing LPLs with dysplasia with OSCC. CD8+ cells were significantly more abundant in OSCC and LPLs with dysplasia compared with LPLs without dysplasia. Proliferating cells (Ki67+) were significantly more abundant in OSCC compared to LPLs with dysplasia. Confocal laser scanning microscopy revealed colocalization of LCs and T cells in LPLs with dysplasia and in OSCC. LCs and T cells are more numerous in tissue compartments with dysplastic epithelial cells and dramatically increase in OSCC. This indicates an ongoing immune response against cells with dysplasia.
Lithofacies and cyclicity of Mohilla evaporite basins on the rifted margin of the Levant in the Late Triassic, Makhtesh Ramon, southern Israel
Authors: BIALIK OR M. , KORNGREEN DORIT , BENJAMINI CHAIM
Published: April 2012

Marine‐connected basins with evaporites occur beneath most extensional continental margins that originated at low‐latitudes and often are of major economic significance. Cyclicity in the evaporite lithofacies reflects the degree of restriction of the basin, overprinted by sea‐level changes, and caused by structural movements in the barrier region, whether by fault‐block rotation, footwall uplift or hanging wall subsidence, in both extensional and compressional basins. The Upper Triassic evaporites of the Ramon section in southern Israel model cyclic sedimentation in such environments. The Mohilla Formation is a carbonate–evaporate–siliciclastic succession of Carnian age that fills a chain of basins extending along the Levant margin from southern Israel to Jordan and Syria. The basins developed in half‐grabens adjacent to normal faults that formed during a period of regional extension. Evaporites of this formation are well‐exposed in outcrops at Makhtesh Ramon, the southernmost of these basins. The M2 Member of the Mohilla Formation is composed of 42 sub‐metre cycles of alternating dolostone, gypsum and calcareous shales. Field and microfacies analysis showed these cycles to conform mostly to restricted shallow and marginal marine environments, spatially limited by the uplifted shoulders of the half‐graben systems. A total of 10 facies types belonging to six depositional environments have been identified. From stacking patterns and analysis of bed to bed change, cycles can be categorized into three groupings: (i) low frequency exposure to exposure cycles that developed under eustatic or climate control; (ii) high frequency deepening/shallowing‐upward cycles, characterized by gradual transitions due to short‐term sea‐level or runoff‐event oscillations possibly referable to orbital forcing; and (iii) high frequency shallowing‐upward cycles, characterized by abrupt transitions, attributable to sporadic tectonic events affecting accommodation space or barrier effectiveness. The way facies and cycling of the sedimentary environments was deciphered in the Mohilla evaporite basin can be used to unravel the genesis of many other evaporite basins with barriers of tectonic origin.
Structural homology in the Solanaceae: analysis of genomic regions in support of synteny studies in tomato, potato and pepper
Authors: Peters Sander A. , Bargsten Joachim W. , Szinay Dóra , van de Belt José , Visser Richard G.F. , Bai Yuling , de Jong Hans
Published: April 2012

We have analysed the structural homology in euchromatin regions of tomato, potato and pepper with special attention for the long arm of chromosome 2 (2L). Molecular organization and colinear junctions were delineated using multi‐color BAC FISH analysis and comparative sequence alignment. We found large‐scale rearrangements including inversions and segmental translocations that were not reported in previous comparative studies. Some of the structural rearrangements are specific for the tomato clade, and differentiate tomato from potato, pepper and other Solanaceous species. Although local gene vicinity is largely preserved, there are many small‐scale synteny perturbations. Gene adjacency in the aligned segments was frequently disrupted for 47% of the ortholog pairs as a result of gene and LTR retrotransposon insertions, and occasionally by single gene inversions and translocations. Our data also suggests that long distance intra‐chromosomal rearrangements and local gene rearrangements have evolved frequently during speciation in the Solanum genus, and that small changes are more prevalent than large‐scale differences. The occurrence of sonata and harbinger transposable elements and other repeats near or at junction breaks is considered in the light of repeat‐mediated rearrangements and a reconstruction scenario for an ancestral 2L topology is discussed.
The mitochondrial genome of Malus domestica and the import‐driven hypothesis of mitochondrial genome expansion in seed plants
Authors: Goremykin Vadim V. , Lockhart Peter J. , Viola Roberto , Velasco Riccardo
Published: April 2012

Mitochondrial genomes of spermatophytes are the largest of all organellar genomes. Their large size has been attributed to various factors; however, the relative contribution of these factors to mitochondrial DNA (mtDNA) expansion remains undetermined. We estimated their relative contribution in Malus domestica (apple). The mitochondrial genome of apple has a size of 396 947 bp and a one to nine ratio of coding to non‐coding DNA, close to the corresponding average values for angiosperms. We determined that 71.5% of the apple mtDNA sequence was highly similar to sequences of its nuclear DNA. Using nuclear gene exons, nuclear transposable elements and chloroplast DNA as markers of promiscuous DNA content in mtDNA, we estimated that approximately 20% of the apple mtDNA consisted of DNA sequences imported from other cell compartments, mostly from the nucleus. Similar marker‐based estimates of promiscuous DNA content in the mitochondrial genomes of other species ranged between 21.2 and 25.3% of the total mtDNA length for grape, between 23.1 and 38.6% for rice, and between 47.1 and 78.4% for maize. All these estimates are conservative, because they underestimate the import of non‐functional DNA. We propose that the import of promiscuous DNA is a core mechanism for mtDNA size expansion in seed plants. In apple, maize and grape this mechanism contributed far more to genome expansion than did homologous recombination. In rice the estimated contribution of both mechanisms was found to be similar.
Antigen persistence of rapid diagnostic tests in pregnant women in Nanoro, Burkina Faso, and the implications for the diagnosis of malaria in pregnancy
Authors: Kattenberg Johanna H. , Tahita Christian M. , Versteeg Inge A. J. , Tinto Halidou , Traoré‐Coulibaly Maminata , Schallig Henk D. F. H. , Mens Petra F.
Published: April 2012

Objectives  To evaluate persistence of several Plasmodium antigens in pregnant women after treatment and compare diagnostics during treatment follow‐up.

Methods  Thirty‐two pregnant women (N = 32) with confirmed malaria infection by a histidine‐rich protein 2 (HRP2)‐based rapid diagnostic test (RDT) and microscopy were followed for 28 days after artemisinin‐based combination therapy (ACT). A Plasmodium lactate dehydrogenase (pLDH)‐based RDT and two ELISAs based on the detection of dihydrofolate reductase–thymidylate synthase (DHFR‐TS) and haeme detoxification protein (HDP) were compared with each other and to RT‐PCR at each visit.

Results  The mean visit number (95% confidence interval) on which the HRP2‐based RDT was still positive after treatment was 3.4 (2.7–4.1) visits with some patients still positive at day 28. This is significantly later than the pLDH‐based RDT [0.84 (0.55–1.1)], microscopy (median 1, range 1–3), DHFR‐TS‐ELISA [1.7 (1.1–2.3)] and RT‐PCR (median 2, range 1–5) (P < 0.05), but not significantly later than HDP‐ELISA [2.1 (1.6–2.7)]. Lower gravidity and higher parasite density at day 0 resulted in significantly longer positive results with most tests (P < 0.05).

Conclusions  HRP2 can persist up to 28 days after ACT treatment; therefore, this test is not suitable for treatment follow‐up in pregnant women and can generate problems when using this test during intermittent preventive treatment (IPTp). DHFR‐TS is less persistent than HRP2, making it a potentially interesting target for diagnosis.
Expression profiles of regulatory and helper T‐cell‐associated genes in nasal polyposis
Authors: Li C. W. , Zhang K. K. , Li T. Y. , Lin Z. B. , Li Y. Y. , Curotto de Lafaille M. A. , Shi L. , Wang D. Y.
Published: April 2012

Background

Nasal polyposis (NP) is a Th2‐skewed inflammatory disorder, but it is unclear what role regulatory T cells (T‐reg) play in disease pathology. We investigated the expression profiles of T‐reg and T‐helper‐cell‐associated genes and their response to glucocorticosteroid (GC) treatment in Chinese patients with NP.

Methods

Biopsies were obtained from 29 non‐treated NP patients for comparison with inferior turbinates collected from healthy controls. In 13 patients, NP samples were collected both before and after short‐term oral GC treatment. Levels of mRNA for T‐cell markers were determined by microarray and quantitative PCR. Cellular infiltrates were assessed by histo‐ and immunohistochemistry.

Results

FOXP3+ T‐reg were increased in GC‐naïve NP, and numbers were negatively correlated with eosinophil infiltration. Helios staining was not detected, suggesting that FOXP3+ cells in NP are not thymus‐derived T‐reg. Compared with controls, mRNA levels corresponding to T‐reg genes were significantly increased in NP (FOXP3, TGFB1, IL10, SMAD3, IL2RA, and JAK3), but transcription factors associated with Th2 (GATA3) or Th17 responses (RORc) were significantly reduced. FOXP3 mRNA levels positively correlated with other T‐reg cell markers. Microarray analysis showed that most Th2‐related markers (e.g., Eotaxin‐1, CCL13, and CCL18) were upregulated in GC‐naïve NP vs controls. GC therapy significantly suppressed eosinophilic inflammation in NP, but did not significantly alter the expression levels of T‐reg/Th2‐associated genes.

Conclusions

Upregulation of FOXP3+‐inducible T‐reg cells and downregulation of Th2 and Th17 markers in NP indicate a regulatory response occurring at a site of persistent mucosal inflammation. However, immune regulation fails to control the underlying tissue pathology. Expression of T‐reg/Th2 markers after GC treatment was unaltered, suggesting that T‐cell‐driving NP inflammatory mediators are GC resistant.