First Report of Umbel Browning and Stem Necrosis Caused by Diaporthe angelicae on Carrot in France.

In 2011, carrot (Daucus carota L.) seed production occurred on 2,900 ha, which accounts for approximately 25% of the area devoted to the production of vegetable fine seeds. Since 2007, symptoms of umbel browning have been regularly observed in carrot production areas located in the central region. Initially, triangular necrotic lesions appeared on carrot umbels that later spread to the entire umbels and often progressed to the stems. Diseased umbels became dried prematurely, compromising seed development. The loss in seed production was estimated at approximately 8% of the harvested carrot umbels during the cropping seasons of spring and summer 2007 and 2008 in France. In collaboration with seed companies, diseased carrot stems were collected from seven fields of seed production (eight plants per field) and a fungus was isolated from the tissue. The cultures were grown on malt (2%) agar (1.5%) medium and incubated for 2 weeks at 22°C in darkness. Young fungal colonies were white and a brownish green pigmentation developed when the colonies became older. The same color was observed from the top and on the reverse of the colonies. To induce sporulation, isolates were grown on water agar (1.5%) medium in the presence of carrot stem fragments for 1 week at 22°C in darkness, followed by 1 week at 22°C in white light under a 16-h photoperiod. Pycnidia were produced on stem fragments and contained alpha and beta conidia typical of the genus Diaporthe (2). Alternatively, pycnidia were also obtained on malt agar medium after 2 weeks of culture at 25°C in white light under a 12-h photoperiod. The size of alpha and beta conidia was 6.3 ± 0.5 × 2.3 ± 0.4 µm and 23.3 ± 1.8 × 0.9 ± 0.2 µm, respectively (n = 170). In order to confirm the identification at the genus level and determine the species, DNA was extracted from the mycelium of three representative isolates and the ITS regions of the ribosomal DNA were amplified using universal primers (1). The sequences of the amplified products (GenBank Accession Nos. KF240772 to KF240774) were 100% identical with the ITS sequence of a Diaporthe angelicae isolate deposited in the NCBI database (CBS 111592 isolate, KC343027). To confirm pathogenicity, the three isolates of D. angelicae were inoculated on carrot umbels in the greenhouse. A total of nine plants were inoculated (three plants per isolate). Using a micropipette, 10 µl of a conidial suspension containing alpha and beta conidia (105 conidia mL-1) were deposited at the base of the primary umbel and two secondary umbels, which were wounded before inoculation using a scalpel blade. Seven inoculated plants developed triangular, necrotic lesions that were typical umbel browning. D. angelicae was re-isolated on malt agar medium from the inoculated diseased carrot umbels. To our knowledge, this is the first report of D. angelicae in carrot cultivated for seed production in France. The disease resembles the lesions described in the Netherlands in 1951 on carrot inflorescence caused by Phomopsis dauci (3). In future experiments, it would be crucial to precisely determine if D. angelicae could be transmitted to the seeds. References: (1) M. A. Innis et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990. (2) J. M. Santos and A. J. L. Philips. Fungal Divers. 34:111, 2009. (3) J. A. von Arx. Eur. J. Plant Pathol. 57:44, 1951.

Activation of natural killer T cells by alpha-galactosylceramide treatment prevents the onset and recurrence of autoimmune Type 1 diabetes.

Type 1 diabetes (T1D) in non-obese diabetic (NOD) mice may be favored by immune dysregulation leading to the hyporesponsiveness of regulatory T cells and activation of effector T-helper type 1 (Th1) cells. The immunoregulatory activity of natural killer T (NKT) cells is well documented, and both interleukin (IL)-4 and IL-10 secreted by NKT cells have important roles in mediating this activity. NKT cells are less frequent and display deficient IL-4 responses in both NOD mice and individuals at risk for T1D (ref. 8), and this deficiency may lead to T1D (refs. 1,6-9). Thus, given that NKT cells respond to the alpha-galactosylceramide (alpha-GalCer) glycolipid in a CD1d-restricted manner by secretion of Th2 cytokines, we reasoned that activation of NKT cells by alpha-GalCer might prevent the onset and/or recurrence of T1D. Here we show that alpha-GalCer treatment, even when initiated after the onset of insulitis, protects female NOD mice from T1D and prolongs the survival of pancreatic islets transplanted into newly diabetic NOD mice. In addition, when administered after the onset of insulitis, alpha-GalCer and IL-7 displayed synergistic effects, possibly via the ability of IL-7 to render NKT cells fully responsive to alpha-GalCer. Protection from T1D by alpha-GalCer was associated with the suppression of both T- and B-cell autoimmunity to islet beta cells and with a polarized Th2-like response in spleen and pancreas of these mice. These findings raise the possibility that alpha-GalCer treatment might be used therapeutically to prevent the onset and recurrence of human T1D.

Interleukin 7 induces preferential expansion of V beta 8.2+CD4-8- and V beta 8.2+CD4+8- murine thymocytes positively selected by class I molecules.

We analyzed the phenotype and V beta-T cell receptor (TCR) repertoire, together with interleukin 7 receptor (IL-7R) expression in unfractionated thymocytes stimulated in vitro with IL-7. This culture system results in a specific proliferation of mature thymocytes belonging to the CD3+CD4-, CD4+8-, and CD4-8+ subsets. IL-7 induced a preferential expansion of V beta 8.2+CD4-8- and V beta 8.2+CD4-8- thymocytes. This phenomenon is not observed in beta 2-microglobulin-deficient mice, showing that a fraction of CD4+8- thymocytes, enriched in V beta 8.2+ cells, is selected by class I molecules in normal mice, as are a large proportion of CD4-8- alpha beta TCR+ thymocytes. Our findings also establish that IL-7 plays a major role in the expansion of rare thymocyte subsets, which could exert important functions in inflammatory and immune responses.

[Diagnostic performance of κ/λ serum free light chain ratio (Freelite® assay) and IgGκ/IgGλ ratio (Hevylite® assay) as prognostic biomarkers of the evolution of immune thrombocytopenia into a chronic disease in adult patients]. / Performance diagnostique des rapports κ/λ des chaines légères libres sériques (test Freelite®) et IgGκ/IgGλ (test Hevylite®) comme marqueurs pronostiques de chronicisation du purpura thrombopénique immunologique de l'adulte.

INTRODUCTION: Immune thrombocytopenia (ITP) is an acquired hemorrhagic disease due to antiplatelet antibodies, that will become a chronic disease in 70% of adults. Most of chronic ITP patients display clonal restriction of antiplatelet antibodies. To date, there is no biomarker able to predict the evolution of the disease. The objective of the study is to determine whether Hevylite® and/or Freelite® assays are prognostic factors for progression to chronic ITP. METHODS: This is a retrospective, monocentric, prognostic study of a biomarker, performed using frozen samples stored in a serum library. Freelite® and a Hevylite® assays were performed on the samples collected at diagnosis for adult patients with newly diagnosed ITP at the University Hospital of Poitiers between 2014/01/01 and 2017/05/01. To predict the evolution into a chronic disease, a ROC curve analysis was performed on four variables: IgGκ, IgGκ/IgGλ ratio, IgGκ - IgGλ, and κ/λ ratio. RESULTS: Thirty-two patients were included and analyzed. No patient had an abnormal κ/λ ratio. Three patients had an abnormal IgGκ/IgGλ ratio. The following variables IgGκ, IgGκ/IgGλ, IgGκ - IgGλ, and κ/λ ratio were not able to predict progression to chronic ITP in our study. CONCLUSION: This study did not reveal any prognostic value of the Freelite® and Hevylite® tests on the evolution of ITP into a chronic disease.

Remobilization of leaf S compounds and senescence in response to restricted sulphate supply during the vegetative stage of oilseed rape are affected by mineral N availability.

The impact of sulphur limitation on the remobilization of endogenous S compounds during the rosette stage of oilseed rape, and the interactions with N availability on these processes, were examined using a long-term (34)SO(4)(2-) labelling method combined with a study of leaf senescence progression (using SAG12/Cab as a molecular indicator) and gene expression of the transporters, BnSultr4;1 and BnSultr4;2, involved in vacuolar sulphate efflux. After 51 d on hydroponic culture at 0.3 mM (34)SO(4)(2-) (1 atom% excess), the labelling was stopped and plants were subject for 28 d to High S-High N (HS-HN, control), Low S-High N (LS-HN) or Low S-Low N (LS-LN) conditions. Compared with the control, LS-HN plants showed delayed leaf senescence and, whilst the shoot growth and the foliar soluble protein amounts were not affected, S, (34)S, and SO(4)(2-) amounts in the old leaves declined rapidly and were associated with the up-regulation of BnSultr4;1. In LS-LN plants, shoot growth was reduced, leaf senescence was accelerated, and the rapid S mobilization in old leaves was accompanied by decreased (34)S and SO(4)(2-), higher protein mobilization, and up-regulation of BnSultr4;2, but without any change of expression of BnSultr4;1. The data suggest that to sustain the S demand for growth under S restriction (i) vacuolar SO(4)(2-) is specifically remobilized in LS-HN conditions without any acceleration of leaf senescence, (ii) SO(4)(2-) mobilization is related to an up-regulation of BnSultr4;1 and/or BnSultr4;2 expression, and (iii) the relationship between sulphate mobilization and up-regulation of expression of BnSultr4 genes is specifically dependent on the N availability.

IL-33 receptor ST2 deficiency attenuates renal ischaemia-reperfusion injury in euglycaemic, but not streptozotocin-induced hyperglycaemic mice.

AIM: Kidney hypoxia can predispose to the development of acute and chronic renal failure in diabetes. Ischaemia-reperfusion injury (IRI) causes inflammation, and diabetes is known to exacerbate this inflammatory response in the kidney, whereas alarmin IL-33 could act as an innate immune mediator during kidney IRI. Thus, the present study examined the impact of genetic IL-33 receptor ST2 deficiency (ST2-/-) on renal IRI in euglycaemic and hyperglycaemic mice. METHODS: Hyperglycaemia was induced with streptozotocin (STZ) in adult male C57BL/6JRj wild-type (WT) mice and ST2-/- mice. Unilateral renal IRI was achieved 3months after STZ treatment by left kidney nephrectomy (non-ischaemic control kidney) and clamping of the right renal artery for 32min in STZ- and vehicle-treated animals. At 24h after reperfusion, renal function and injury were determined by levels of plasma creatinine, blood urea nitrogen (BUN) and histological tubule scores. Also, in a complementary pilot clinical study, soluble ST2 concentrations were compared in diabetics and non-diabetics. RESULTS: Urinary albumin was significantly increased in STZ-induced hyperglycaemic mice, regardless of genotypic background. At 24h post-ischaemia, plasma creatinine, BUN and tubular injury were significantly reduced in ST2-/- mice compared with vehicle-treated WT mice, but this protective effect was lost in the STZ-induced hyperglycaemic ST2-/- animals. Plasma concentrations of soluble ST2 were significantly greater in type 2 diabetes patients vs non-diabetics. CONCLUSION: Our data suggest that the IL-33/ST2 pathway exerts differential effects depending on the glucose environment, opening-up new avenues for future research on alarmins and diabetes in ischaemia-related diseases.

Modulation of human lymphocyte proliferation by FLFQPQRFamide, a FMRFamide-like peptide with anti-opiate properties.

The octapeptide Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2 (F8Fa), originally detected in mammalian brain by antisera raised against the invertebrate peptide Phe-Met-Arg-Phe-NH2 (FMRFamide) is a neuropeptide able to antagonize the actions of both endogenous and exogenous opiates. Since it is well accepted that lymphocytes are targets for opiates, we have tested the effect of F8Fa on T cell proliferation from normal human peripheral blood lymphocytes. Our study shows that F8Fa exerts a concentration-dependent diphasic modulation of human T lymphocyte proliferation. Thus, despite a great variability between individuals, 10(-13) M F8Fa was found to enhance the proliferation of T cells induced by phytohemagglutinin or anti-CD2 monoclonal antibodies, while 10(-7) M F8Fa inhibited T cell proliferation, without affecting cell viability. When F8Fa was tested on monocyte-depleted cell preparations, only the inhibitory effect was observed. These results indicate that F8Fa may stimulate T cells via monocytes, but may also directly inhibit T lymphocyte proliferation. Given the presence of F8Fa-like peptide in human plasma, we suggest that F8Fa may act as a neurohormone in the control of the immune system.

Characterization of binding sites for neuropeptide FF on T lymphocytes of the Jurkat cell line.

Neuropeptide FF (NPFF) is a neuropeptide with antiopiate properties able to antagonize the action of both endogenous and exogenous opiates. Because we have recently shown that NPFF modulates the proliferation of human T lymphocytes, we have searched for binding sites for this peptide on T lymphocytes. Our study shows that T lymphocytes of the Jurkat cell line express binding sites for [125I]YLFQPQRFamide, an iodinated analogue of NPFF. This binding is time and dose dependent, reversible, saturable, and may be resolved in two distinct components of high and low affinity. The opiate receptor agonists mu, delta, and kappa, as well the antagonist naloxone, were unable to affect binding. Beside the effects of opiates on immune cells, our results suggest that an antiopiate peptide, such as NPFF, could play a role in the modulation of the immune system.

Identification of the B cell derived T cell colony promoting activity to soluble CD23.

We previously reported that supernatants of phytohemagglutinin (PHA)-stimulated normal human B cells (NBCsup) contain a T cell colony promoting activity. NBCsup were able to (a) increase the number of secondary colonies generated under PHA and interleukin-2 (IL-2) stimulation by peripheral blood-derived primary T colony cells, (b) enhance the ability of CD4+ but not CD8+ peripheral blood T cells to form agar colonies in the presence of PHA and IL-2 and (c) support in vitro differentiation of CD2-3-4-8- prothymocytes into CD2+3+4+ T cells. This activity was therefore refered to as Prothymocyte Differentiating Activity (PTDA). Subsequent studies pointed to striking biochemical and cell source homologies between this B cell derived factor and the 25-kDa soluble CD23 (sCD23). sCD23 has been recently found to display prothymocyte differentiating activity.

How do illiterate adults react to metalinguistic training?

The present study focuses on the capacity of illiterate adults to master three different metalinguistic tasks: judgment of phonological length of words, initial consonant deletion, and lexical segmentation of sentences. Illiterates' performance, during a pre-test and after training, was compared with that of literates and partial illiterates (adults at the beginning of the process of acquiring literacy) who received the same training. In the pre-test, illiterates were lower than literates in the three tasks; and partial-illiterates were at an intermediate level in two of the tasks. The three groups profited from the training, especially illiterates and partial-illiterates for whom improvement was dramatic across all three tasks. Finally, the results revealed a hierarchy of difficulty across the tasks. The capacity to focus on the phonological dimension seemed to be a prerequisite for the phoneme deletion ability. The task of lexical segmentation seemed to be more a measure of syntactic awareness than a measure of phonological awareness.

C-reactive protein levels at the onset of labour and at day 3 post-partum in normal pregnancy.

OBJECTIVES: To record maternal serum C-reactive protein levels during normal onset of labour and normal puerperium and to evaluate if inflammation or infection could be predicted during these two periods when serum C-reactive protein is increased. METHODS: Eighty-five pregnant women were enrolled in a longitudinal prospective study and had a blood sample to assess serum C-reactive protein levels on admission to the labour ward for normal onset of labour and at day three post-partum. Inclusion criteria were no previous history, a normal single pregnancy, normal vaginal delivery and an uneventful post-partum course. Twelve non-pregnant women of the same age constitued a control group. An automatic Behring Nephelometer was used to measure serum C-reactive protein concentrations. The Student's t-test (significance p < 0.05) was used for statistical analysis. FINDINGS: C-reactive protein was significantly increased during the onset of labour (4.10 +/- 2.79 mg/L) and reached very high levels during the post-partum period (24.07 +/- 18.28 mg/L) compared to the standard normal serum C-reactive protein level in a population of non-pregnant women of the same age (2.39 +/- 0.07 mg/L). INTERPRETATION: Increased serum C-reactive protein has been reported to be a marker for subclinical infection during pregnancy in various situations including premature labour and premature rupture of membranes and for complications occurring during puerperium such as thrombophlebitis, thromboembolism or endometritis. This interpretation depends on which upper limit is considered as abnormal. Because serum C-reactive protein was raised during the onset of labour, values of less than 10 mg/L could not be considered as a marker for infection during this period. Elevated serum concentrations of estrogen, progestogen and prostaglandins during labour might be one explanation for those physiological changes. Normal vaginal delivery could be compared to a surgical procedure and tissue injury consecutive to vaginal birth as reflected by a dramatic increase in C-reactive protein. More studies using nephelometry are needed to determine normal and upper values of C-reactive protein during pregnancy.

Do deaf children use phonological syllables as reading units?

This study aimed at examining whether deaf children process written words on the basis of phonological units. In French, the syllable is a phonologically and orthographically well-defined unit. French deaf children and hearing children matched on word recognition level were asked to copy written words and pseudo-words. The number of glances at the item, copying duration, and the locus of the first segmentation (i.e., after the first glance) within the item were measured. The main question was whether the segments copied by the deaf children corresponded to syllables as defined by phonological and orthographic rules.The results showed that deaf children, like hearing children, used syllables as copying units when the syllable boundaries were marked both by orthographic and phonological criteria. However, in a condition in which orthographic and phonological criteria were differentiated, the deaf children did not perform phonological segmentations while the hearing children did. We discuss two explanatory hypotheses. First, items in this condition were difficult to decode for deaf children; second, orthographic units were probably easier to process for deaf children than phonological units because of a lack of automaticity in their phonological conversion processes for pseudo-words. Finally, incidental observations during the experimental task raised the question of the use of fingerspelled units.

Identification of invariant natural killer T cells in porcine peripheral blood.

The pig is a relevant preclinical model for numerous pathologies used to validate therapeutic strategies for translation to human. Although invariant natural killer T (iNKT) lymphocytes are a component of innate immunity implicated in many pathological processes, little is known on their characterization in swine. By addressing this issue using mouse α-galactosylceramide-loaded CD1d tetramers (α-GC-CD1dTT), which are commonly used to track iNKT cells, we were able to unequivocally identify CD3(+)α-GC-CD1dTT(+) cells in porcine peripheral blood, hereafter referred to as swine iNKT cells. These lymphocytes are enriched in CD4(-)CD8(+) and CD4(-)CD8(-) cells, harbor an activated-memory phenotype (SLA-DR(+)CD45RA(-)), express the intracellular promyelocytic-leukemia-zinc-finger (PLZF) transcription factor and are significantly enriched in IFN-γ-producing cells after in vitro activation in comparison with conventional T cells. Importantly, in presence of IL-2 and IL-15, the iNKT cell ligand α-GC induces selective expansion of CD3(+)α-GC-CD1dTT(+) cells, confirming the reactivity of swine iNKT cells against α-GC. When associated with α-GC, IL-33, an alarmin of IL-1 family recently described to target iNKT cells, leads to a greater expansion of CD3(+)α-GC-CD1dTT(+) cells than IL-2 and IL-15. Altogether, our results provide the first phenotypic and functional description of swine iNKT cells allowing to further study the critical role of iNKT cells in porcine models of organ injury.