Articles de revues sur le sujet « Concordata suspensiva »

A candidin, which is a suspension of killed yeast cells, is commonly used for intradermal tests of delayed hypersensitivity, to evaluate the immunological cellular competence of the patient, when the test is applied along with other similar tests. When working with a cellular antigen, the histopathology of positive skin tests reveals a cellular infiltrate which not only presents a characteristic hypersensitivity reaction but also a neutrophilic abscess in the central part. This research presents the results of a comparison between the yeast cell suspension and the polysaccharide antigens, both obtained from the same strains of Candida albicans. The results obtained by skin tests in one hundred individuals were 61.0% with the polysaccharide antigen and 69.0% with the yeast cell suspension antigen. Concordant results concerning the two antigens were observed in 82.0% of the individuals. The discussion section presents an assumption to explain the differences of positivity obtained with the two antigens. We conclude that the polysaccharide antigen can be utilized in the intradermal test of delayed hypersensitivity to Candida albicans.

A novel oligonucleotide suspension microarray (Luminex microsphere system) was developed for the rapid detection of avian respiratory viruses of major clinical importance. This test was optimized and validated with 70 clinical samples. The developed tool was accurate for high-throughput detection and differentiation of the most important avian respiratory viruses: avian influenza virus (AIV), Newcastle disease virus (NDV), infection bronchitis virus (IBV), and infectious laryngotracheitis virus (ILTV) in single- and mixed-virus infections. A multiplex reverse transcriptase PCR (RT-PCR), followed by a monoplex or a multiplex Luminex assays, were realized using a Luminex 200 analyzer instrument. The sensitivity, specificity, and reproducibility of the multiplex DNA suspension microarray system were evaluated. The results showed no significant differences in the median fluorescence intensity (MFI) value in monoplex and multiplex Luminex assays. The sensitivity and specificity proved to be completely concordant with monoplex real-time RT-PCR. We demonstrated that the multiplex DNA suspension microarray system is an accurate, high-throughput, and relatively simple method for the rapid detection of the main respiratory viruses of poultry.

Objective: The present study was designed to investigate the anti-stress activity of Cassia auriculata ethanolic seed extract in mice. Methodology: The anti-stress effect was evaluated by using Elevated plus maze (EPM), Force swimming test (FST) and Tail suspension test (TST). The ECS at a doses (250,500 and 1000 mg/kg p.o.) and standard (diazepam 2 mg/kg i.p and fluoxetine 20 mg/kg i.p) was administered. Results: The extract showed the increased in the number of entries and time spent in open arm in Elevated plus maze and decreased in the immobility time in both Force swimming test and Tail suspension test. Conclusion: The effect of ESC on animal behavior was concordant with a significant regulation of GABA and stress hormones. Therefore, this study was attempted to demonstrate the preventive potential of ECS against stress disorders at in vivo levels

El presente trabajo plantea un análisis de los temas eclesiásticos que fueron objeto de discusión en el primer Congreso constituyente de Michoacán de 1824 a 1825. En concreto, la investigación se centra en los debates y resoluciones que se dieron en torno a los honores a las autoridades civiles, los diezmos, la presencia de los eclesiásticos en el Congreso estatal y los asuntos eclesiásticos dentro del debate constitucional en 1825. Las discusiones se desarrollaron en un momento en que no existía un marco legal sólido por la falta de reconocimiento de la independencia de México por España y la Santa Sede y la suspensión del Patronato hasta un nuevo concordato; lo que permite ver que los conflictos entre el poder civil y religioso se hicieron presentes a pocos días de haber abierto sesiones el constituyente michoacano.

This study aimed to investigate the antistress properties of the ethanol extract ofCymbopogon schoenanthus(CSEE), growing wild in the southern part of Tunisia. The effect of extracts on H2O2-induced cytotoxicity and stress in human neuroblastoma SH-SY5Y cells. Its effect on stress-induced in ICR mice was exposed to force swim and tail suspension, in concordance with heat shock protein expression (HSP27 and HSP90), corticosterone, and catecholamine neurotransmitters level. Our results demonstrated that pretreatment of SH-SY5Y cells with CSEE at 1/2000, 1/1000, and 1/500 v/v dilutions significantly inversed H2O2-induced neurotoxicity. Moreover, CSEE treatments significantly reversed heat shock protein expression in heat-stressed HSP47-transformed cells (42°C, for 90 min) and mRNA expression of HSP27 and HSP90 in H2O2-treated SH-SY5Y. Daily oral administration of 100 mg/kg and 200 mg/kg CSEE was conducted to ICR mice for 2 weeks. It was resulted in a significant decrease of immobility time in forced swimming and tail suspension tests. The effect of CSEE on animal behavior was concordant with a significant regulation of blood serum corticosterone and cerebral cortex levels of catecholamine (dopamine, adrenaline, and noradrenaline). Therefore, this study was attempted to demonstrate the preventive potential of CSEE against stress disorders atin vitroandin vivolevels.

High frequencies of somaclonal phenotypic and cytogenetic variation have been observed previously among regenerants from cotton (Gossypium hirsutum L., 2n = 4x = 52). In this study we endeavored to determine if cytogenetic abnormalities would be detectable in cotton cell cultures and if so, whether or not the observed abnormalities would parallel those expected on the basis of previous cytogenetic analyses of cotton somaclones. Paired samples from suspension cultures established from 21-month-old 'Coker 312' and 8-month-old 'Coker 315' calli were pretreated or not pretreated with colchicine to detect cytogenetic abnormalities at metaphase or anaphase–telophase, respectively. Cell cultures established from both calli were found to vary in chromosome number. Hypoaneuploidy was common, but hyperaneuploidy and polyploidy were rare. Modal chromosome numbers for the 'Coker 312' and 'Coker 315' cultures were 46 and 50, respectively. Bridges at anaphase and telophase were frequent in the 'Coker 312' cultures but rare in the 'Coker 315'cultures. Cytogenetic differences between the cultures could be due to effects of culture age, genotype, their interaction, or other factors. Very small chromosomes, presumably centric fragments, as well as ring chromosomes and putative bridges between metaphase chromosomes occurred at low frequencies. The prevalence of hypoaneuploidy and rarity of hyperaneuploidy and polyploidy in cultures paralleled previous results on cotton somaclones, indicating that cytogenetic abnormalities arising in vitro probably contribute significantly to cotton somaclonal variation. The occurrence of hypoaneuploidy and bridges, including multiple bridges within single cells, is concordant with the hypothesis that breakage–fusion–bridge cycles may accumulate during in vitro culture of cotton.Key words: cotton, Gossypium, tissue culture, cytogenetics.

Abstract The Hologic Panther Fusion® Open Access™ functionality allows implementation of laboratory-developed tests (LDTs), with fully automated sample extraction, real-time PCR, and result interpretation. We report the development and validation of a multiplex LDT for norovirus G1, norovirus G2, and rotavirus from stool samples on this system. The LDT was optimized for primer and probe sequences, salt concentration, and PCR annealing temperature. Reproducibility of the PCR and extraction process was assessed. Performance of the multiplex LDT assay was evaluated with external quality assessment (EQA) samples and compared to a commercial multiplex assay (Allplex™ GI-Virus Assay, Seegene) in clinical samples. Salt concentrations and annealing/extension temperature were optimized to 4 mM MgCl2, 70 mM KCl, 20 mM Tris, and 60 °C, respectively. The user-prepared part of the LDT PCR mix (containing salts, probes, and primers) was stable for ≥ 11 days onboard the instrument. We observed reproducible results of PCR and the extraction process. The LDT had a sensitivity comparable to or greater than the commercial Allplex™ assay and showed excellent linearity. Forty-five EQA samples yielded the expected result with the LDT. There was 100% concordance between LDT and Allplex™ results in 160 clinical samples. Results from the suspension and direct swab stool sample preparation methods were highly concordant in the LDT. We report the successful development and validation of a multiplex PCR LDT for detection of norovirus G1, norovirus G2, and rotavirus from stool samples on the Panther Fusion® system.

Blueberries (Vaccinium corymbosum) have recently become an important alternative crop in different ecological regions of Argentina. In surveys, a new disease characterized by leaf spots and twig and shoot blight has been observed on plants cultivated in Arrecifes, Mercedes, and San Pedro (provinces of Buenos Aires) and Concordia (province of Entre Ríos) since July 2004. Spots initially appear brown, circular, 1 to 2 mm in diameter, and irregularly distributed on the leaves and they eventually coalesce. Fruiting twig and shoot blight developed from the tips toward the base. Affected plants of cvs. O'Neal and Reveille were distributed randomly in the field and with a low incidence (average of 2%). The objective of this work was to identify the causal agent of this disease. Symptomatic plant material was surface disinfested with 0.2% NaOCl for 1 min and 70% ethanol for 1 min, washed once with sterile distilled water, blotted dry with paper towels, and plated on potato dextrose agar. Colonies were initially white, becoming light to dark gray with the onset of sporulation with black, sphaerical to subsphaerical conidia that measured 14 to 19 × 12 to 16 μm. These characteristics agree with published descriptions of Nigrospora sphaerica (Sacc.) Mason (1,4). To evaluate pathogenicity, all leaves, petioles, and stems of seven healthy potted plants of cv. O'Neal were punctured with flamed needles and sprayed with a suspension of 1 × 108 spores of the fungus per milliliter of sterile distilled water. Another seven nonwounded plants were sprayed with the spore suspension. Seven plants similarly injured and seven nonwounded plants were sprayed with sterile distilled water and served as controls. Each plant was covered with a water-sprayed polyethylene bag and maintained in a controlled environment chamber at 20°C with a 12-h photoperiod. The bags were removed after 3 days. All wounded inoculated plants began to show disease symptoms similar to those observed in the field 20 days after inoculation. Controls and nonwounded inoculated plants remained symptomless. The pathogen was reisolated from diseased tissues fulfilling Koch's postulates. N. sphaerica is a well-known saprophyte on many plant species but has been mentioned as pathogen on many hosts (2,3). To our knowledge, this is the first reference of N. sphaerica as a wound pathogen of blueberry. In the field, the fungus would have gained access to the plant through wounds caused by insects or frost after a long-term wetness duration. References: (1) M. B. Ellis. Dematiaceous Hyphomycetes. CMI, Kew, Surrey, UK, 1971. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. American Phytopathological Society, St. Paul, MN, 1989. (3) D. F. Farr et al. Fungal Databases. Systematic Botany and Mycology Laboratory. Online publication. ARS, USDA. 2007. (4) E. W. Mason. Trans. Brit. Mycol. Soc. 12:152, 1927.

Abstract A new method has been developed to evaluate leukemias based on binding of cells in suspension to a microarray of cluster of differentiation (CD) monoclonal antibodies (mAbs) immobilized on nitrocellulose film on a microscope slide. Cell binding is proportional to antigen expression. This method allows simultaneous evaluation of 88 different CD antigens and is feasible with homogeneous populations of mononuclear cells obtained from blood of patients with leukemia. This methodology is potentially useful in diagnosis and classification of different leukemias and lymphomas and it may further aid in identifying and distinguishing subgroups of patients within a particular diagnosis. We utilized this methodology to assess its effectiveness in distinguishing prognostic subgroups in patients with chronic lymphocytic leukemia (CLL), those with mutated versus unmutated IgVH genes and those with <20% ZAP70+ versus >20% ZAP70+ cells. Unmutated IgVH genes were defined as >98% homologous to germline. To do this we identified 101 unselected samples from CLL patients for which IgVH mutational status and/or ZAP70 expression were known. Both IgVH status and ZAP70 expression were known for 70, in 6 only IgVH status, and in 25 only ZAP70 was known. IgVH mutational status and ZAP70 expression were concordant in 62/70 cases. Patient characteristics (median and range) were as follows: age=59yrs(37–82); abs lym=23k/μL(3.6–123); β2M=2.3mg/L(1.2–9.5); # prior treatments= 0(0–6). 20 had Rai low-risk; 67 had Rai-intermediate-risk, and 14 had Rai high-risk disease; 78 were previously untreated. Reduced space linear discrimination analysis and empirical Bayes moderated t-test was used to evaluate relationships between microarray binding and patients with “good prognosis” (IgVH mutated or ZAP70-) and “poor prognosis” (IgVH unmutated or ZAP70+). Empirical Bayes analysis was used to identify CD antigens with significant differential expression between the “good prognosis” and “poor prognosis” groups. Although there was not a significant difference in overall expression between the two groups, there were antibodies that had significantly different levels of binding. For the IgVH unmutated group, increased binding was noted with CD95, CD38, CD2, CD13, CD11a, CD86, CD25, CD9, CD22 (p<.05; FDR-adjusted). For the ZAP70+ group, increased binding was noted for CD38, CD2, CD95, CD49d, CD79b, kappa, and CD11a (pFDR<.05). In the IgVH/ZAP70 concordant cases, the following had increased binding with the “poor prognosis” group (IgVH unmutated/ZAP70+): CD38, CD95, CD2, CD13, CD24, CD9, and CD11a (pFDR<.05). There is clear evidence of differential binding on the mAb microarray between prognostic groups, although it is not yet possible to accurately predict ZAP70 or IgVH mutation status based on microarray results. Work continues to correlate these findings with flow cytometry and with other meaningful prognostic factors such as β2M; Rai stage, etc. In addition, follow-up continues to assess correlations between microarray results and significant clinical endpoints including survival. This is a simple immunomicroarray method that may be useful and important to enhance the classification and prognostic assessment of patients with lymphoid malignancies, including CLL, based on correlations between an extensive immunophenotype and clinical endpoints.

Since 2003, a new field disease has been observed on several cultivars of highbush blueberry (Vaccinium corymbosum L.) in Buenos Aires (Baradero, Colonia Urquiza, Lima, Mercedes, and San Pedro), Entre Ríos (Concordia, Gualeguaychú, and Larroque), and Córdoba (Capilla del Monte and La Cumbre). Infected flowers turned brown to tan with a water-soaked appearance and shriveled up. Blighted flowers typically did not produce fruits; even an entire cluster of berries could be aborted. A chlorotic area, that later became necrotic and turned light brown, developed when leaves were in contact with blighted flowers. A watery rot developed on fruit occasionally before harvest but more generally after harvest. Infected tender green twigs also became blighted, with leaf tissue becoming brown to black. Older twigs and stems were also blighted. Abundant, gray mycelium with conidial masses developed on all affected tissues under moist conditions. Sections of infected leaves, twigs, stems, flowers, and fruits were surfaced sterilized with 0.2% NaOCl, plated on 2% potato dextrose agar (pH 7), and incubated at 22°C. Pure cultures formed a whitish dense mycelial mat and turned gray after 72 h. Conidia were ellipsoid, hyaline, nonseptate, and formed on botryose heads. They ranged from 5.8 to 9 × 8.1 to 13.7 μm (average 8.6 × 10.2 μm). Black, round, and irregular microsclerotia developed on 7-day-old cultures with an average size of 1.1 × 1.7 mm. Morphological characteristics agree with those described for Botrytis cinerea Pers.:Fr (1). Pathogenicity was tested on 10 12-month-old potted blueberry plants cv. O'Neal by spraying a suspension of 1 × 106 conidia per ml of sterile distilled water. Ten plants used as controls were sprayed with sterile distilled water. Each plant was covered with a transparent polyethylene bag for 48 h and incubated at 20 ± 2°C in humid chambers for 15 days. Lesions similar to those observed in the fields developed after 4 days and asexual fructifications developed after 5 days. The same pathogen was reisolated from the lesions, thus completing Koch's postulates. Water-treated plants remained symptomless. To our knowledge, this is the first report of a disease caused by B. cinerea on blueberry in Buenos Aires, Córdoba, and Entre Ríos provinces of Argentina. References: (1) M. V. Ellis and J. M. Waller. Sclerotinia fuckeliana (conidial state: Botrytis cinerea) No. 431 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1974.

Abstract Angioimmunoblastic T-cell lymphoma (AITL) is a distinct clinicopathological entity among peripheral T-cell lymphoma in the WHO classification. Whereas antigen “loss” or “deletion” of one or several pan-T cell antigens is a hepful feature of neoplastic lymphocytes in many T-cell lymphomas, no specific immunophenotypic patterns were available to recognize the tumour T-cells of AITL until recently. Indeed, Attygalle et al. reported that in this disorder neoplastic T-cells can be recognized by the aberrant expression of CD10 using immunochemistry in lymph nodes as well as in the involved extranodal sites. Lee et al. has also confirmed this specific phenotypic feature in cell suspension of lymph nodes using flow cytometry (FCM) in 3 cases of AITL. Here, we evaluated the CD10 expression by T cells in patients with AITL using four-colour FCM. The present study included lymph nodes (LN, n=10), peripheral blood (PB, n=5), bone marrow (n=1) and skin (n=1) samples from 13 patients with a diagnosis of AITL and with available cytologic histologic, immunologic and molecular data. Lymph nodes of reactive hyperplasia (n=13), B-cell lymphoma (n=23), other T-cell lymphoma (n=6) and peripheral blood from healthy donors (n=18) were used as control group. According with previous immunohistochemistry results, a fraction of T-cells expressed CD10 (using a level of at least 5% of all CD5+ cells) in 9/10 AITL lymph nodes with a mean number of 18%. Interestingly, among these 9 cases, 5 could be studied in peripheral blood also and all cases showed a fraction of T-cells expressing CD10, whatever be the lymphocytosis (median 1.1 109/l range 0.82 to 11.32 109/l). In three of these cases, tumoral T-cells presented also lack of surface CD3. In two cases of AITL diagnosed in LN, the aberrant CD10 expression by T-cells was found in bone marrow and skin, respectively. In the control group, T-cells were CD10 negative using the cut-off of 5%. In conclusion, we demonstrate that the assessment of CD10 expression by neoplastic T-cells can be achieved by multi-colour FCM in lymph nodes and involved extranodal sites. Our results are concordant with the statement of Attygalle that CD10 expression by T-cells can be used as a marker of both malignancy and AITL type. In addition, this is to our knowledge the first description of circulating CD10 neoplastic T-cells in AITL. Further study with a larger series of patients is required to confirm these data, to standardize the cut-off of positivity and to evaluate the sensibility of FCM versus immunohistochemistry. Figure Figure

Background: A cost effective and comprehensive genomic profiling (CGP) approach for diagnosis, risk stratification and therapy would be useful for the evaluation of oncologic specimens. Available approaches involving additive testing for DNA and RNA abnormalities through traditional methods (e.g. Sanger, FISH, cytogenetics, qRT-PCR) are not comprehensive, require multiple different workflows and are sample consuming, often resulting in incomplete testing. While there are next generation sequencing (NGS) assays designed for detecting DNA and RNA abnormalities, they have separate workflows that require twice the amount of sample and effort. To address this, we developed a novel total nucleic acid (TNA) extraction method and single tube workflow utilizing TNA and a custom multimodal chemistry designed for hematologic malignancies. This consolidated workflow enables an efficient discovery based approach for both DNA/RNA abnormalities including single nucleotide variants (SNVs), InDels, copy number variants (CNVs), large structural changes from DNA and gene fusions and gene expression levels from RNA. This method maximizes data derived from valuable samples while delivering a comprehensive profile of the patient's tumor which can help guide therapeutic and clinical decisions. Methods: Total nucleic acid (TNA) was extracted from bone marrow and peripheral blood of 95 patients (CML, CMML, CLL, AML and myeloid disorders). 297 genes that have DNA mutations specific to hematological cancers were targeted, along with 213 genes that were targeted for clinically significant RNA abnormalities. Enriched genomic and transcriptomic regions of interest from 85 patients were successfully sequenced with unique dual indices on an Illumina NovaSeq 6000. DNA variant detection as well as fusion detection from RNA were compared to traditional orthogonal NGS assays that use DNA input or compared to qRT-PCR and Sanger sequencing assays that use RNA as input. Results: In this study, we developed an efficient and high-quality TNA extraction method that can purify enough total nucleic acid from bone marrow, peripheral blood, cytogenetic pellets, flow suspension, and FFPE samples for the downstream NGS assay. The average OD 260/280 value was 1.9 and the OD 260/230 was 2.18. After sequencing, 256/262 (97.7% accuracy) SNV and Indel variants that were candidate pathogenic mutations were concordant from 38 patients. Meanwhile, 100% (7/7) of all BCR/ABL1 gene fusions which had an international scale (IS) value above 6.4% were concordant. In addition, 69 fusion positive samples containing 20 unique gene fusions which had been previously reported by an independent ArcherDX assay designed specifically for gene fusions were also evaluated with this chemistry. Analysis revealed a 92.5% (64/69) concordance. More importantly, the QIAseq multimodal TNA NGS assay detected both DNA and RNA abnormalities in a single tube. For example, in one myeloid leukemia patient, we not only identified pathogenic variants of ASXL1 and JAK2 which had been previously detected by a DNA NGS assay, but also detected a concurrent BCR-FGFR1 fusion which had been previously reported by a FISH assay. Moreover, we were able to provide more comprehensive genomic profiling by investigating many DNA and RNA abnormalities simultaneously. In our study, for 5 patients that previously been tested for BCR-ABL1 fusion only, we are able to assess BCR-ABL1 fusion status from RNA as well as identify pathogenic DNA variants at the same time, including JAK2 p.V617F, U2AF1 p.S34F, ASXL1 p.E635Rfs*15, BRCA p.S1982Rfs*22, and DNMT3A p.S708Vfs*71, which provides valuable information to assist diagnosis and treatment in a cost effective and efficient way. Conclusions: We developed a single tube TNA based workflow with a custom multimodal chemistry that simultaneously detects many DNA and RNA abnormalities in a cost effective and efficient way while reducing sample requirements. This unique TNA NGS assay provides comprehensive genomic profiling for hematologic malignancies and improves the diagnostic testing options for precise patient care. Disclosures Yu: NeoGenomics: Current Employment. Alarcon:NeoGenomics: Current Employment. Mou:NeoGenomics: Current Employment. Jung:NeoGenomics: Current Employment. Nam:NeoGenomics: Current Employment. Thomas:NeoGenomics: Current Employment. Keeler:NeoGenomics: Current Employment. Shinbrot:NeoGenomics: Current Employment. Magnan:NeoGenomics: Current Employment. Bender:NeoGenomics: Current Employment. Jiang:NeoGenomics: Current Employment. Agersborg:NeoGenomics: Current Employment. Weiss:Bayer: Other: speaker; Genentech: Other: Speaker; Merck: Other: Speaker; NeoGenomics: Current Employment. Ye:NeoGenomics: Current Employment. Funari:NeoGenomics: Current Employment.

Abstract Abstract 2265 Background: Tissue factor (TF) is essential for the hemostatic initiation of the extrinsic pathway of blood coagulation. We found that cancer and benign tumor cells have different TF-dependent procoagulant activities (PCA). Intravascular microparticle-, cancer-, or other cell-associated TF may initiate thrombosis but circulating TF levels are not necessarily concordant with thrombosis. We investigated the role of TF carrier burden on the procoagulant activity of circulating TF. Methods: We utilized human monocytic-like U937 cells and thromboplastin-coated polymer microspheres as TF carriers. U937 cells were stimulated with endotoxin to induce surface expression of TF. Particle effect on clotting was measured in a closed transport system using a coagulometer. Occlusive thrombus formation was measured in an open transport system using a flow chamber under a constant pressure gradient. Zymogen activation was recorded with a chromogenic substrate in a closed transport system. Results: This study was designed to assess whether the spatial separation of intravascular TF-carriers in blood, demonstrated with TF-inducible human monocytic cell line U937 or TF-coated polymer microspheres, affected procoagulant activity and hence thrombogenic potential. Experiments were performed to characterize the effects of TF carrier number on the kinetics of clot formation in both open and closed systems. The procoagulant activity of TF carriers was found to correlate with spatial separation in both closed, well-mixed systems as well as open, flowing systems. TF carriers enhanced the amidolytic activity of FVIIa towards the chromogenic substrate, S-2366, as a function of carrier count. Initiation time correlated with spatial separation, whereas enzyme reaction rates correlated with inverse spatial separation. Conclusions: These findings suggest that transport of coagulation factors to the surface of a circulating TF carrier may constrain the enzymatic rate of TF-dependent coagulation reactions, and therefore modulate circulating TF procoagulant activity. By shortening the separation of TF carriers in suspension, the proximity of plasma coagulation factors to a TF surface is increased, and we measured a resulting increase in procoagulant activity for these carriers. Our results suggest that patient-specific sensitivity to circulating TF may depend upon plasma concentrations of TF-dependent coagulation factors (FVII, FX and FII) as the concentration of factors in plasma influences their transport rate. Further, the distribution of circulating TF within the vasculature may influence procoagulant activity of circulating TF by altering the proximity of TF-carriers to plasma coagulation factors. Thus, clinical relevance of circulating TF may not depend on whole blood TF concentration, but rather on the distribution of circulating TF over different carriers (tumor cells versus tumor cell-derived microparticles) or patient-specific concentration of plasma coagulation factors in the extrinsic pathway (i.e. FVII) downstream of TF. Disclosures: Gruber: Aronora, LLC: Consultancy, Equity Ownership.

Introduction: Multiple myeloma (MM) is characterized by expansion of neoplastic plasma cells (PCs). Disease presentation is heterogenous with hypercalcemia, renal failure, osteolytic lesions or a combination thereof. The overall survival of patients and their response to treatment has improved considerably however, MM stays incurable. Patients develop treatment resistance and progress to relapse. Disease progression to treatment resistance and relapse are attributed to complex signaling molecules. In vitro experiments on cell lines are pivotal to determine the functional characterization of these molecules leading to identification of potential therapeutic targets. Cell lines have served as instruments for research in varied disciplines including cancer. Currently, majority of the cell lines in cancer research are from European origin and to a lesser extent from Africans, Asians and Hispanic populations. This lacuna of inadequate racial and genetic representation leads to major consequences such as delays in the benefits of precision medicine, drug efficacy and partial understanding of molecular aspects of the disease. The study was undertaken to establish and characterize a cell line from a patient diagnosed with MM. Methods AKU-MY01 was established from a 38-year male patient diagnosed with ISS stage III, kappa light chain myeloma in 2013. Plasma cells were isolated from the BM aspirate using sterile conditions and cultured in RPMI-1640 supplemented with 10% FBS. Conditioned media from urinary bladder carcinoma cell line 5637 was used to provide growth factors. Isolated suspension cells were observed in culture which were subcultured by splitting in a 1:3 ratios. Detailed characterization of AKU-MY01 was undertaken using gene expression analysis, population doubling time (PDT), determination of clonality, fluorescent in situ hybridization, immunocytochemistry (ICC), karyotyping, transwell assay and short tandem repeat (STR) analysis. Results: AKU-MY01 showed expression of ABCG2dim, Oct4, CD38, CD138, CD19, CD20, CD45, CD56 dim, XBP1, MUC1, EBNA1, and monoclonal Lambda light chain. AKU-MY01 exhibited a human diploid karyotype with 46XY and IGH deletions/translocation in 20% of the cells. It has a PDT of 48-53 hrs. and high invasive potential (15.6%) compared to other MM cell lines. Expression of EBNA at mRNA level was further confirmed through fluorescent ICC. The results from Immunofluorescent ICC showed clusters of tumor cells with concordant expression of membranous CD138 and cytosolic and nuclear expression of LMP1. Further, the expression of LMP1 in CD138 positive tumor cells was confirmed in Patient's trephine from whome AKU-MY01 was derived. Conclusion: To the best of our knowledge, AKU-MY01 is the first MM cell line characterized for LMP1 expression in CD138 expressing myeloma cells. Salient features of AKU-MY01 which make it a unique addition to the existing repertoire of MM cell lines are a) Asian origin, b) diploid karyotype and c) expression of EBV protein and mRNA. Further studies using this cell line model may contribute to understand the biology of EBV positive MM cases. Disclosures No relevant conflicts of interest to declare.

From 2006 to 2009, crown gall and hairy root symptoms were observed on blueberry (Vaccinium corymbosum cvs. O'Neil, Millennia, and Misty) plants from six nurseries in Tucumán, Concordia, Pilar, Morón, and Baradero, Argentina. Bacteria were isolated from galls of all three cultivars and from hairy roots of Millenia and O'Neil onto D1 and D1M agar media at 27°C. Typical Agrobacterium colonies developed in 5 days (2). Seven bacterial strains (five from galls and two from hairy roots) were studied further. All were gram negative, aerobic, and catalase positive with rod-shaped cells that synthesized β–galactosidase and metabolized D-glucose, D-arabinose, n-acetyl-glucosamine, maltose, mannitol, and malonate. Strains were negative for lysine decarboxylase, H2S production, indole, and 3-ketolactose production. While gall strains were urease positive and citrate variable (mostly positive), hairy root strains were urease negative, citrate positive, had poly-β-hydroxybutyrate inclusion granules, and clarified acid on potato dextrose agar containing 0.5% CaCO3 (2). Agrobacterium tumefaciens ATCC 15955 and LBA 958 were included as controls. PCR with virA/C primers amplified a 338-bp product corresponding to the virD2 operon and confirmed that the strains harbored a pathogenic plasmid (1). Bacterial strains were assigned to biovars with a multiplex PCR assay targeting 23S rRNA sequences (3). Two strains produced PCR amplicons typical of A. rhizogenes bv. 2. The other five strains produced PCR amplicons typical of A. rubi, which were insensitive to agrocin in a bioassay with A. radiobacter strain K1026. Identity was confirmed by sequencing the 16S rDNA of strains F 266 (GenBank No. GU580894) and F 289 (No. GU580895), which had 99% homology to 16sRNA sequences of A. rubi ICMP 11833 (AY626395.1) and A. rhizogenes ATCC 11325 (AY945955.1), respectively. Pathogenicity of all seven strains was tested on V. corymbosum cv. Misty, Bryophyllum daigremontiana, tobacco cv. Xanthi, tomato cv. Presto, and pepper cv. California Wonder. Plants were inoculated by a needle stabbed into the stems with the appropriate cell suspension (108 CFU/ml) of each strain or with sterile distilled water (control treatment). Two plants of each species were tested per strain. Plants were grown for at least 45 days at 23 ± 3°C and symptoms were recorded. Inoculations with the five strains isolated from galls caused development of spherical, white to flesh-colored, rough, spongy wart-like galls at the inoculation sites. Root strains induced root proliferation on all inoculated plants as well as in a carrot disk bioassay (4). On blueberry plants, galls were dark brown to black, rough, and woody 6 months after inoculation. No lesions were observed on control plants. Bacteria were reisolated from symptomatic tissues of inoculated plants. Enterobacterial repetitive intergeneric consensus-PCR confirmed that the DNA fingerprints of the reisolated strains were identical to those of the original strains. To our knowledge, this is the first report of A. rubi and A. rhizogenes causing hairy root and crown gall on blueberry in Argentina. References: (1) J. H. Haas et. al. Appl. Environ. Microbiol. 61:2879,1995. (2) L. W. Moore et al. Page 17 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. N. W. Schaad et al., eds. The American Phytopathological Society, St. Paul, MN, 2001. (3) J. Pulawska et al. Syst. Appl. Microbiol. 29:470, 2006. (4) M. H. Ryder et al. Plant Physiol. 77:215, 1985.

Moldy core is a fungal disease of apple fruits that is characterized by mycelial growth in the seed locules and is sometimes accompanied by penetration of the immediate surrounding flesh. The disease can go undetected until the fruit is cut open, as no external symptoms appear on the fruit. Alternaria, Aspergillus, Cladosporium, Coniothyrium, Epicoccum, Phoma and Stemphylium are some of the common pathogens associated with moldy core (Serdani et al. 2002; Gao et al. 2013; McLeod 2014). The disease is more common in apple cultivars with an open calyx, where spores may initiate infections during the growing season or at the post-harvest storage stage (Spotts et al. 1988). In 2018, a shipment of ‘Sweet Tango’ apples from New Zealand to Scotian Gold Co-operative Ltd., Nova Scotia, Canada, was found to be affected by moldy core. Moderate to severe moldy core symptoms were observed when 10 apples were cut open (Figure S1). In comparison, ‘Sweet Tango’ apples grown in Nova Scotia showed no moldy core symptoms when 10 random fruits were cut open. Small pieces of the diseased fruit tissue from the core region were surface-disinfected for 1 min in 1% NaOCl, rinsed three times with sterilized water and placed onto potato dextrose agar (PDA) dishes. The PDA dishes were incubated in dark at 22 oC and single spore isolation was carried out to fresh PDA dishes. These isolate produced colonies of regular shape, tan black with prominent white gray margin and gray colour conidia (Figure S2 AB). The colonies turn dark black after 3 weeks of growth on PDA. Mycelia were septate and conidia were oval or obclavate or club-shaped with a tapering end with 4-6 longitudinal and transverse septa (Figure S2 C-D). The size of conidia ranges from 12.5-20 x 8.7-12.5 µM on 20 days old PDA dishes. Based on the size and shape of conidia and other morphological characteristics the isolated fungi were identical to Alternaria spp. (Simmons 2007). To assess the identity of the isolated pathogen species by multi-locus sequence analysis, genomic DNA was extracted from the pure cultures of two isolates (5.8 and 8) using the E.Z.N.A. SP Fungal DNA Kit (Omega Bio-Tek). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH), major allergen (Alt a 1), OPA10-2, the internal transcribed spacer (ITS) region of ribosomal DNA and the translation elongation factor 1-α (TEF1-α) region from two Alternaria spp. isolates (5.8 and 8) were amplified and sequenced using primers gpd1/2 (Berbee et al. 1999), A21F/A21R (Gabriel 2015), OPA10-2/ OPA10-2L (Andrew et al. 2009), ITS1/ITS4 (White et al. 1990) and EF1-up /EF1-low (O’Donnell et al. 1998) respectively. The resulting sequences of both isolates were deposited in the NCBI GenBank (GAPDH; MW411052, MW411053, Alt a 1; MW411050, MW411051, OPA10-2; MW415762, MW415763, ITS; MK140445, MT225559, TEF1-α; MT305773 and MT305774 ). Sequences of GAPDH, Alt a 1, OPA-10-2, ITS and TEF1-α genes of both isolates were identical to each other and showed 100 %, 100 %, 99.21 %, 100% and 100% identity to A. arborescens S. (AY278810.1, AY563303.1, KP124712.1, KY965831.1, KY965831.1) respectively. Identity with reference strain CBS 102605 confirms that both of the isolated strains 5.8 and 8 are A. arborescens. The pathogenicity of the two A. arborescens isolates were confirmed by artificially inoculating healthy ‘Sweet Tango’ fruit by dispensing the conidial suspension directly on the seed locule. Briefly, surface-disinfected fruits were air-dried for 5 min and then peeled using a sterilized knife and cut transversally. Each half of the fruit was inoculated with 100 µl of conidial suspensions (∼1 × 104 conidia/ml) in potato dextrose broth (PDB) and incubated at 22 °C in a humid chamber for 7–10 days, or until symptoms with visible mycelial growth were observed. The control fruits were treated with 100 µl of sterilized PDB. Both A. arborescens isolates produced visible moldy core symptoms on the inoculated ‘Sweet Tango’ fruits, whereas no symptoms were observed on the control fruits (Figure S1). The experiment was repeated three times with at least three replicates with similar results. A. arborescens was successfully re-isolated from the artificially-inoculated fruits to complete Koch’s postulates. To our knowledge, this is the first report of Alternaria arborescens causing moldy core disease in ‘Sweet Tango’ apples from New Zealand. Acknowledgments We thank Eric Bevis for his help in sample preparation for DNA sequencing, Willy Renderos for pathogenicity assay. We also thank Joan Hebb (Scotian Gold Cooperative Ltd.,) for providing the apple sample for this study. This research was made possible through financial support from Agriculture and Agri-Food Canada. The authors(s) declare no conflict of interest. Literature Cited Andrew M., Peever T.L., Pryor B.M. An expanded multilocus phylogeny does not resolve species among the small-spored Alternaria species complex. 2009. Mycologia. 101:95–109. Berbee, M. L. et al. 1999. Cochliobolus phylogenetics and the origin of known, highly virulent pathogens, inferred from ITS and glyceraldehyde-3-phosphate dehydrogenase gene sequences Mycologia. 91:964. Gabriel, M.F. I. Postigo, A. Gutiérrez-Rodríguez, E. Suñén, C.T. Tomaz, J. Martínez 2015. Development of a PCR-based tool for detecting immunologically relevant Alt a 1 and Alt a 1 homologue coding sequences. Medical Mycology. 53 (6):636–642. Gao, L. L., Zhang, Q., Sun, X. Y., Jiang, L., Zhang, R., Sun, G. Y., Zha, Y. L., and Biggs, A. R. 2013. 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