Null Type Research Articles

Abstract NTOC-093: SYNTHETIC LETHAL APPROACHES TO TARGET ARID1A DEFICIENT OVARIAN CANCERS

Abstract Epithelial ovarian cancer (EOC) remains the most lethal gynaecological malignancy in the Western world. Ovarian clear cell carcinoma (OCCC), a distinct histological subtype, has a notably poor prognosis in the advanced setting compared to patients with high-grade serous ovarian cancer (HGSOC). Understanding why these patients have a poor outcome may be due to the underlying genetic drivers and their response to treatment. Dysregulation of the SWI/SNF complex is one of the most commonly occurring defects in solid cancers. Mutations in ARID1A (AT-rich interactive domain-containing protein 1A), a gene that encodes for BAF250A, forming part of the SWI/SNF chromatin remodeling complex, rarely occur in HGSOC but are common in ovarian clear cell carcinomas. The vast majority of these are loss of function frameshift or nonsense mutations, resulting in loss of protein function. In addition, loss of ARID1A expression in tumour specimens has been associated with a shorter progression free survival and chemoresistance in ovarian clear cell carcinoma (OCCC). Despite the understanding that ARID1A defects are associated with tumourigenesis, targeted therapy approaches that exploit this deficiency have not as yet been developed. Our aims were to identify ways of targeting ARID1A deficient tumours by performing a large-scale functional genomics screen to identify actionable synthetic lethal effects. Using a high-throughput combination drug screen with a plate library of 80 compounds and a phase 1 compound, in isogenic ARID1A null and wild type HCT116 cells, we have identified candidate therapeutic approaches to targeting ARID1A mutant tumours that could be assessed in proof of concept clinical trials. We have undertaken subsequent high throughput drug screens in isogenic ARID1A null and wild type MCF10A cells that in we have identified a series of novel synthetic lethal effects. Assessment of this combinatorial approach in in vivo models of ARID1A mutant cancers is now underway. In conclusion, we have identified clinically actionable combinatorial approaches that may provide additional therapeutic benefit for ARID1A deficient patients. Citation Format: Saira Khalique, Chris T. Williamson, Helen Pemberton, Patty T. Wai, Malini Menon, Rachel Brough, Andri Leonidou, Barrie Peck, Susana Banerjee, Rachael C. Natrajan and Christopher J. Lord,. SYNTHETIC LETHAL APPROACHES TO TARGET ARID1A DEFICIENT OVARIAN CANCERS [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr NTOC-093.

Null hypothesis significance testing and Type I error: The domain problem

Abstract Although many common uses of p-values for making statistical inferences in contemporary scientific research have been shown to be invalid, no one, to our knowledge, has adequately assessed the main original justification for their use, which is that they can help to control the Type I error rate (Neyman & Pearson, 1928, 1933). We address this issue head-on by asking a specific question: Across what domain, specifically, do we wish to control the Type I error rate? For example, do we wish to control it across all of science, across all of a specific discipline such as psychology, across a researcher's active lifetime, across a substantive research area, across an experiment, or across a set of hypotheses? In attempting to answer these questions, we show that each one leads to troubling dilemmas wherein controlling the Type I error rate turns out to be inconsistent with other scientific desiderata. This inconsistency implies that we must make a choice. In our view, the other scientific desiderata are much more valuable than controlling the Type I error rate and so it is the latter, rather than the former, with which we must dispense. But by doing so—that is, by eliminating the Type I error justification for computing and using p-values—there is even less reason to believe that p is useful for validly rejecting null hypotheses than previous critics have suggested.

GATA3 expression and its relationship with clinicopathological parameters in invasive breast carcinomas

GATA3, as a transcription factor, is associated with estrogen receptor (ER) expression and necessary for luminal cell differentiation in mammary glands. Association of GATA3 expression with clinicopathological parameters, molecular subtypes of tumors, disease free survival (DFS) and overall survival (OS) for breast carcinoma patients were evaluated in this study. We immunohistochemically stained GATA3, CK5/6, EGFR, CK14 and vimentin on tissue microarray blocks of 457 invasive breast carcinomas. Tumors are sub-classified as luminal A, luminal B, HER2 expressing, basal-like and null type according to their hormonal status with cerbB2, CK 5/6 and EGFR expressions. Follow-up data for 254 cases were obtained. 215/457 (47%) tumors were GATA3 positive. GATA3 expression was inversely correlated with mitotic count (p<0.0001), nuclear grade (p=0.001), histological grade (p=0.001), tumor necrosis (p=0.001), stromal lymphocytic response (p<0.01), nipple invasion (p=0.01), metastasis (p=0.03), vimentin (p=0.0003), EGFR (p=0.015) and CK14 (p=0.001) expressions; and directly associated with ER (p<0.0001) and progesterone receptor (PR) (p<0.0001) expressions. Luminal A carcinomas had the highest frequency for GATA-3 (140/245), however basal-like carcinomas had the lowest (1/42) (p<0.0001). None of the medullary and metaplastic carcinomas expressed GATA3. GATA3 was associated with good DFS and OS (p=0.001 and p=0.0009) and was an independent prognostic factor for OS. GATA3 expression, regardless of the subtype, may have a prognostic significance for breast carcinomas through its ability to promote the differentiation of luminal progenitor cells.

Investigation of mucosal pattern of gastric antrum using magnifying narrow-band imaging in patients with chronic atrophic fundic gastritis.

BackgroundMagnifying narrow-band imaging (M-NBI) can reportedly help predict the presence and distribution of atrophy and intestinal metaplasia in the gastric corpus. However, the micro-mucosal pattern of the antrum shown by M-NBI differs from that of the corpus. We studied the distribution and histology of the micro-mucosal pattern in the antrum based on magnifying endoscopy.MethodsEndoscopic images of the greater curvature of the antrum were evaluated in 50 patients with chronic atrophic fundic gastritis (CAFG). The extent of CAFG was evaluated by autofluorescence imaging. The micro-mucosal pattern was evaluated by M-NBI and classified into groove and white villiform types. The localization of white villiform type mucosa was classified into three types in relation to the areae gastricae: null, central, and segmental types. Biopsies were taken from regions showing different micro-mucosal patterns. Associations among the extent of CAFG, micro-mucosal pattern, and histology were examined.ResultsAs the extent of CAFG increased, the proportion of white villiform type mucosa increased, whereas that of groove type mucosa decreased (P=0.022). In patients with extensive CAFG, most of the areae gastricae was composed of the segmental or central type of white villiform type mucosa (P=0.044). The white villiform type mucosa had significantly higher grades of atrophy (P=0.002) and intestinal metaplasia (P<0.001) than did the groove type mucosa.ConclusionWhite villiform type mucosa is indicative of atrophy and intestinal metaplasia in the gastric antrum. It extends to the whole or central part of the areae gastricae as CAFG becomes more extensive.

Nrf2 Modulates Host Defense during Streptococcus pneumoniae Pneumonia in Mice

Nrf2 regulates the transcriptional response to oxidative stress. These studies tested the role of Nrf2 during Streptococcus pneumoniae pneumonia and identified Nrf2-dependent genes and pathways in lung tissue and in recruited neutrophils. Nrf2 null and wild type (WT) mice were studied at 6 and 24 h after instillation of S. pneumoniae or PBS. At 6 h, fewer neutrophils were recruited and the number of bacteria remaining in the lungs tended to be less (p = 0.06) in the Nrf2 null compared with WT mice. In uninfected lungs, 53 genes were already differentially expressed in Nrf2 null compared with WT mouse lungs, and gene sets involved in phagocytosis, Fc receptor function, complement, and Ig regulation are enhanced in PBS-treated Nrf2 null gene profiles compared with those of WT mice. These results suggest that initial host defense is enhanced in Nrf2 null mice, resulting in less recruitment of neutrophils. At 24 h, neutrophil recruitment was greater. The percentages of early apoptotic and late apoptotic/necrotic neutrophils were similar. At increasing inoculum numbers, mortality rates strikingly increased from 15 to 31 and 100% in Nrf2 null mice, whereas all WT mice survived, and Nrf2 null mice had a defect in clearance, particularly at the intermediate dose. The mortality was due to enhanced lung injury and greater systemic response. Gene profiling identified differentially regulated genes and pathways in neutrophils and lung tissue, including those involved in redox stress response, metabolism, inflammation, immunoregulatory pathways, and tissue repair, providing insight into the mechanisms for the greater tissue damage and increased neutrophil accumulation.

Abstract 1857: Dysregulation of transsulfuration enzymes contribute to malignant transformation in a murine model of colitis-associated carcinogenesis

Abstract Introduction: Ulcerative colitis (UC) is a highly morbid, chronic inflammatory disease characterized by mucosal ulceration of the colonic mucosa and is associated with the higher risk of colitis-associated cancer (CAC). The exact mechanism(s) causing UC progression to CAC is currently unknown. The balanced activity of transsulfuration pathway enzymes, cystathionine-gamma-lyase (CSE) and cystathionine-beta-synthase (CBS), and their production of endogenous hydrogen sulfide gas, are critical to maintenance of the colonic homeostasis. CSE activity is suggested to be important for wound healing and mucosal protection. Consistent with this, we have previously showed a decrease CSE expression in human colonic mucosa obtained from patients with chronic UC, compared to normal colonic mucosa by immunocytochemistry. By contrast, increased CBS expression is implicated in the progression of sporadic colorectal carcinoma. However, the role of these enzymes in CAC is unknown. We hypothesize that the dysregulation in CSE/CBS expression and activity is important to the progression from UC to CAC. Methods: CSE null mice and wild type Sv129/B6 (control) mice were used in azoxymethane-dextran sodium sulfate (AOM-DSS) colon cancer model which mimics human CAC. The disease development was followed up to day 80. Confocal microscopy and Western blot was used to assess the gene expression during cancer development. Size, number, and time interval to tumor formation, as well inflammation were assessed. Results: Abrogation of CSE expression using CSE null animals in AOM-DSS model of CAC accelerated the time to tumor development and resulted in an increase in both tumor size (p&amp;lt;0.001) and number (p&amp;lt;0.001) compared to wild-type controls. CBS protein expression was increased within the colonic tumor when compared to the normal margin in AOM-DSS treated animals by Western blot analysis and tissue immunostaining. Interestingly, CAC liver metastases, an exceedingly rare finding in this mouse model, were identified. Conclusion: Taken together, our human and murine data suggest that dysregulation in the transsulfuration pathway enzymes CSE and CBS expression/activity may be critical contributor to the CAC development in UC and may serve as a potential biomarker for disease progression in the future. Citation Format: Paul Johnson, Ches’Que M. Phillips, Carl Grim, John R. Zatarain, Aakash H. Gajjar, Suimin Qiu, Rui Wang, Celia Chao, Iryna V. Pinchuk, Mark R. Hellmich. Dysregulation of transsulfuration enzymes contribute to malignant transformation in a murine model of colitis-associated carcinogenesis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1857.

550 - In vitro and in vivo testing of a novel DNA methyl transferase inhibitor (DNMTI) with activity against hematogenous and solid tumors whether p53 null or wild type (WT)

550 - In vitro and in vivo testing of a novel DNA methyl transferase inhibitor (DNMTI) with activity against hematogenous and solid tumors whether p53 null or wild type (WT)

Expression patterns of GATA3 and the androgen receptor are strongly correlated in patients with triple-negative breast cancer

GATA-binding protein 3 (GATA3) is a diagnostically useful immunohistochemical marker of breast cancer. Because of its strong association with estrogen receptor expression, GATA3 has markedly reduced sensitivity in triple-negative breast cancer (TNBC). We constructed a tissue microarray using a large series of TNBCs and evaluated GATA3 expression by TNBC subtype as defined by surrogate immunohistochemical markers. A total of 205 TNBCs were classified into cancers of the molecular apocrine type (n=23, 11.2%), claudin-low type (n=21, 10.2%), basal-like type (n=91, 44.4%), mixed type (n=62, 30.2%), and null type (n=8, 3.9%). The GATA3 scores (staining intensity × proportion) were categorized as negative (0), focally positive (1-10), or positive (11-300). GATA3 staining was negative in 153 cancers (74.6%), focally positive in 11 (5.4%), and positive in 41 (20.0%). The rate of focal positivity or positivity for GATA3 was significantly higher in the molecular apocrine type (73.9%, 17/23) than in other types of TNBCs (P=.001). The mean GATA3 score of molecular apocrine-type TNBC was significantly higher than that of the other types (P=.001) and differed significantly between androgen receptor (AR)-positive and AR-negative TNBCs (P<.001). In conclusion, GATA3 expression was correlated strongly with AR-positive, molecular apocrine-type TNBCs. Co-expression of AR and GATA3 is a specific feature of molecular apocrine-type TNBC, which may serve as a diagnostic aid for cancer of unknown primary.

The epithelial-mesenchymal transition phenotype of metastatic lymph nodes impacts the prognosis of esophageal squamous cell carcinoma patients.

Epithelial-mesenchymal transition (EMT) plays a key role in tumor metastasis, but the significance of EMT phenotype to the prognosis of esophageal squamous cell carcinoma (ESCC) patients remains unclear. We used immunohistochemistry to examine the expression of the EMT-related proteins E-cadherin, N-cadherin and vimentin in samples of T3N1-3M0 ESCC from 155 primary tumors (PTs) with paired metastatic lymph nodes (MLNs) and 58 PTs without paired MLNs. Based on the expression pattern of the EMT markers, PTs and MLNs were classified as EMT wild, hybrid, null or complete type. The hybrid (42.7%) and complete (39.4%) types predominated among PTs, whereas the wild (34.2%) and hybrid (52.9%) types predominated among MLNs, and EMT phenotypes differed between the paired PTs and MLNs (P < 0.001). Univariate analysis revealed that, for PTs, the EMT phenotype was associated with N-stage (P = 0.039) but not patient survival, and that patients with complete or hybrid type MLNs had better overall survival (OS, P = 0.001) and disease-free survival (DFS, P = 0.005) than patients with null and wild type MLNs, especially those with N1-stage disease (P = 0.017 for OS, and P = 0.017 for DFS, respectively). Multivariate analysis revealed that wild and null type MLNs as well as older age and N2-3 stage were independent predictors of OS and DFS (P < 0.05). Thus MLNs exhibit EMT phenotypes that are distinct from those of their PT and may serve as a novel independent prognostic indicator in ESCC.

Large-cell lung carcinoma by the 2015 WHO classification: a clinicopathologic analysis of 93 cases

To analyze the clinicopathologic characteristics of large-cell lung carcinoma (LCC) subtypes according to the 2015 WHO classification. A total of 93 cases of LCC were identified among 1 634 cases of morphologically undifferentiated non-small cell lung carcinoma. Clinicopathologic and immunohistochemical characteristics were retrospectively reviewed. Among 93 cases of LCC, 50 cases were solid adenocarcinoma (53.8%), 21 cases were non-keratinzing squamous cell carcinoma (22.6%), 3 cases were adenosquamous carcinoma (3.2%), 8 cases were large cell carcinoma with unclear immunohistochemical features (8.6%), and 11 cases were large cell carcinoma with null immunohistochemical features (11.8%). No significant differences in clinical features were found among various immunophenotypes. The overall survival of 1, 3 and 5 years of all cases was 51.0%, 5.4% and 1.1%, respectively and the mean survival was (15.2±1.2)months. Notably the null type had the lowest survival rate of 18% in one year, significantly worse than those of solid adenocarcinoma and non-keratinzing squamous cell carcinoma. The 2015 WHO classification based on immunohistochemical phenotype replaces the morphological approach in separating poorly differentiated non-small cell lung carcinoma from the former large cell lung carcinoma. The null immunohistochemical subtype may represent a distinct subtype of undifferentiated carcinoma with the worst prognosis.

Absence of TNF-α enhances inflammatory response in the newborn lung undergoing mechanical ventilation.

Bronchopulmonary dysplasia (BPD), characterized by impaired alveolarization and vascularization in association with lung inflammation and apoptosis, often occurs after mechanical ventilation with oxygen-rich gas (MV-O2). As heightened expression of the proinflammatory cytokine TNF-α has been described in infants with BPD, we hypothesized that absence of TNF-α would reduce pulmonary inflammation, and attenuate structural changes in newborn mice undergoing MV-O2 Neonatal TNF-α null (TNF-α(-/-)) and wild type (TNF-α(+/+)) mice received MV-O2 for 8 h; controls spontaneously breathed 40% O2 Histologic, mRNA, and protein analysis in vivo were complemented by in vitro studies subjecting primary pulmonary myofibroblasts to mechanical stretch. Finally, TNF-α level in tracheal aspirates from preterm infants were determined by ELISA. Although MV-O2 induced larger and fewer alveoli in both, TNF-α(-/-) and TNF-α(+/+) mice, it caused enhanced lung apoptosis (TUNEL, caspase-3/-6/-8), infiltration of macrophages and neutrophils, and proinflammatory mediator expression (IL-1β, CXCL-1, MCP-1) in TNF-α(-/-) mice. These differences were associated with increased pulmonary transforming growth factor-β (TGF-β) signaling, decreased TGF-β inhibitor SMAD-7 expression, and reduced pulmonary NF-κB activity in ventilated TNF-α(-/-) mice. Preterm infants who went on to develop BPD showed significantly lower TNF-α levels at birth. Our results suggest a critical balance between TNF-α and TGF-β signaling in the developing lung, and underscore the critical importance of these key pathways in the pathogenesis of BPD. Future treatment strategies need to weigh the potential benefits of inhibiting pathologic cytokine expression against the potential of altering key developmental pathways.

TGFβ regulates Galectin-3 expression through canonical Smad3 signaling pathway in nucleus pulposus cells: implications in intervertebral disc degeneration

Galectin-3 is highly expressed in notochordal nucleus pulposus (NP) and thought to play important physiological roles; however, regulation of its expression remains largely unexplored. The aim of the study was to investigate if TGFβ regulates Galectin-3 expression in NP cells. TGFβ treatment resulted in decreased Galectin-3 expression. Bioinformatic analysis using JASPAR and MatInspector databases cross-referenced with published ChIP-Seq data showed nine locations of highly probable Smad3 binding in the LGALS3 proximal promoter. In NP cells, TGFβ treatment resulted in decreased activity of reporters harboring several 5′ deletions of the proximal Galectin-3 promoter. While transfection of NP cells with constitutively active (CA)-ALK5 resulted in decreased promoter activity, DN-ALK5 blocked the suppressive effect of TGFβ on the promoter. The suppressive effect of Smad3 on the Galectin-3 promoter was confirmed using gain- and loss-of-function studies. Transfection with DN-Smad3 or Smad7 blocked TGFβ mediated suppression of promoter activity. We also measured Galectin-3 promoter activity in Smad3 null and wild type cells. Noteworthy, promoter activity was suppressed by TGFβ only in wild type cells. Likewise, stable silencing of Smad3 in NP cells using sh-Smad3 significantly blocked TGFβ-dependent decrease in Galectin-3 expression. Treatment of human NP cells isolated from tissues with different grades of degeneration showed that Galectin-3 expression was responsive to TGF-β-mediated suppression. Importantly, Galectin-3 synergized effects of TNF-α on inflammatory gene expression by NP cells. Together these studies suggest that TGFβ, through Smad3 controls Galectin-3 expression in NP cells and may have implications in the intervertebral disc degeneration.

Matrix metalloproteinase-20 mediates dental enamel biomineralization by preventing protein occlusion inside apatite crystals

Reconstruction of enamel-like materials is a central topic of research in dentistry and material sciences. The importance of precise proteolytic mechanisms in amelogenesis to form a hard tissue with more than 95% mineral content has already been reported. A mutation in the Matrix Metalloproteinase-20 (MMP-20) gene results in hypomineralized enamel that is thin, disorganized and breaks from the underlying dentin. We hypothesized that the absence of MMP-20 during amelogenesis results in the occlusion of amelogenin in the enamel hydroxyapatite crystals. We used spectroscopy and electron microscopy techniques to qualitatively and quantitatively analyze occluded proteins within the isolated enamel crystals from MMP-20 null and Wild type (WT) mice. Our results showed that the isolated enamel crystals of MMP-20 null mice had more organic macromolecules occluded inside them than enamel crystals from the WT. The crystal lattice arrangements of MMP-20 null enamel crystals analyzed by High Resolution Transmission Electron Microscopy (HRTEM) were found to be significantly different from those of the WT. Raman studies indicated that the crystallinity of the MMP-20 null enamel crystals was lower than that of the WT. In conclusion, we present a novel functional mechanism of MMP-20, specifically prevention of unwanted organic material entrapped in the forming enamel crystals, which occurs as the result of precise amelogenin cleavage. MMP-20 action guides the growth morphology of the forming hydroxyapatite crystals and enhances their crystallinity. Elucidating such molecular mechanisms can be applied in the design of novel biomaterials for future clinical applications in dental restoration or repair.

MMP‐9 null mice display elevated IOP due to reduced aqueous humor drainage from the trabecular meshwork

PurposeAqueous humor outflow resistance depends on a complex equilibrium of extracellular matrix biosynthesis versus proteolysis in the trabecular meshwork. Several members of the MMP family, including MMP‐9, are essential to the regulation of aqueous humor outflow resistance, i.e. to intraocular pressure (IOP) homeostasis. Here, we characterized the baseline ocular phenotype of MMP‐9 null mice, with emphasis on the dysregulation of IOP homeostasis.MethodsMMP‐9 null and wild type mice, from 3 to 13 months of age, were studied. IOP and central corneal thickness were measured via rebound tonometry and pachymetry, resp. Anterior chamber morphology and trabecular meshwork organization were studied (1) in vivo with OCT, (2) via light microscopy and (3) by means of transmission electron microscopy (TEM), and its collagen composition was studied using Sirius Red and immunostainings. Integrity of the retina and optic nerve were evaluated with OCT and histological stainings on tissue sections and retinal flatmounts.ResultsMMP‐9 null mice present with early‐onset ocular hypertension, and fluorophotometric measurements of aqueous humor turnover revealed a reduced aqueous humor drainage. While OCT, light microscopy and TEM analysis did not disclose any abnormalities in the cellular organization of the trabecular meshwork, collagen staining indicated that there is an aberrant extracellular matrix composition in MMP‐9 null mice. Remarkably, the observed IOP elevation in MMP‐9 null mice did not result in a glaucomatous phenotype at the level of the retina and optic nerve at the ages studied.ConclusionsOur observations corroborate the role of MMP‐9 as an important remodeler of the trabecular meshwork, and evidence for a causal link between MMP‐9 deficiency, trabecular meshwork composition and IOP elevation is revealed.

Statistics and accuracy of magnetic null identification in multispacecraft data

AbstractComplex magnetic topologies are ubiquitous in astrophysical plasmas. Analyzing magnetic nulls, regions of vanishing magnetic field, is one way to characterize 3‐D magnetic topologies. Magnetic nulls are believed to be important in 3‐D reconnection and turbulence. In the vicinity of a null, plasma particles become unmagnetized and can be accelerated to high energies by electric fields. We present the first statistical study of the occurrence of magnetic nulls and their types in the Earth's nightside magnetosphere. We are able to identify the nulls both in the tail and in the magnetopause current sheets. On average, we find one null for every few current sheet crossings. We show that the type identification of magnetic nulls may be sensitive to local fluctuations in the magnetic field. We develop and demonstrate a method to estimate the reliability of the magnetic null type identification.

CD8+ T Cell Response to Gammaherpesvirus Infection Mediates Inflammation and Fibrosis in Interferon Gamma Receptor-Deficient Mice.

Idiopathic pulmonary fibrosis (IPF), one of the most severe interstitial lung diseases, is a progressive fibrotic disorder of unknown etiology. However, there is growing appreciation for the role of viral infection in disease induction and/or progression. A small animal model of multi-organ fibrosis, which involves murine gammaherpesvirus (MHV68) infection of interferon gamma receptor deficient (IFNγR-/-) mice, has been utilized to model the association of gammaherpesvirus infections and lung fibrosis. Notably, several MHV68 mutants which fail to induce fibrosis have been identified. Our current study aimed to better define the role of the unique MHV68 gene, M1, in development of pulmonary fibrosis. We have previously shown that the M1 gene encodes a secreted protein which possesses superantigen-like function to drive the expansion and activation of Vβ4+ CD8+ T cells. Here we show that M1-dependent fibrosis is correlated with heightened levels of inflammation in the lung. We observe an M1-dependent cellular infiltrate of innate immune cells with most striking differences at 28 days-post infection. Furthermore, in the absence of M1 protein expression we observed reduced CD8+ T cells and MHV68 epitope specific CD8+ T cells to the lungs—despite equivalent levels of viral replication between M1 null and wild type MHV68. Notably, backcrossing the IFNγR-/- onto the Balb/c background, which has previously been shown to exhibit weak MHV68-driven Vβ4+ CD8+ T cell expansion, eliminated MHV68-induced fibrosis—further implicating the activated Vβ4+ CD8+ T cell population in the induction of fibrosis. We further addressed the role that CD8+ T cells play in the induction of fibrosis by depleting CD8+ T cells, which protected the mice from fibrotic disease. Taken together these findings are consistent with the hypothesized role of Vβ4+ CD8+ T cells as mediators of fibrotic disease in IFNγR-/- mice.

J -kink domain walls and the DBI action

We study $J$-kink domain walls in $D=4$ massive $\mathbb{C}P^1$ sigma model. The domain walls are not static but stationary, since they rotate in an internal $S^1$ space with a frequency $\omega$ and a momentum ${\bf k}$ along the domain wall. They are characterized by a conserved current $J_\mu = (Q,{\bf J})$, and are classified into magnetic ($J^2 < 0$), null ($J^2=0$), and electric ($J^2 > 0$) types. Under a natural assumption that a low energy effective action of the domain wall is dual to the $D=4$ DBI action for a membrane, we are lead to a coincidence between the $J$-kink domain wall and the membrane with constant magnetic field $B$ and electric field ${\bf E}$. We also find that $(Q, {\bf J}, \omega, {\bf k})$ is dual to $(B, {\bf E}, H, {\bf D})$ with $H$ and ${\bf D}$ being a magnetizing field and a displacement field, respectively.

How to find magnetic nulls and reconstruct field topology with MMS data?

AbstractIn this study, we apply a new method—the first‐order Taylor expansion (FOTE)—to find magnetic nulls and reconstruct magnetic field topology, in order to use it with the data from the forthcoming MMS mission. We compare this method with the previously used Poincare index (PI), and find that they are generally consistent, except that the PI method can only find a null inside the spacecraft (SC) tetrahedron, while the FOTE method can find a null both inside and outside the tetrahedron and also deduce its drift velocity. In addition, the FOTE method can (1) avoid limitations of the PI method such as data resolution, instrument uncertainty (Bz offset), and SC separation; (2) identify 3‐D null types (A, B, As, and Bs) and determine whether these types can degenerate into 2‐D (X and O); (3) reconstruct the magnetic field topology. We quantitatively test the accuracy of FOTE in positioning magnetic nulls and reconstructing field topology by using the data from 3‐D kinetic simulations. The influences of SC separation (0.05~1 di) and null‐SC distance (0~1 di) on the accuracy are both considered. We find that (1) for an isolated null, the method is accurate when the SC separation is smaller than 1 di, and the null‐SC distance is smaller than 0.25~0.5 di; (2) for a null pair, the accuracy is same as in the isolated‐null situation, except at the separator line, where the field is nonlinear. We define a parameter ξ ≡ |( λ1 + λ2 + λ3 )|/|λ|max in terms of the eigenvalues (λi) of the null to quantify the quality of our method—the smaller this parameter the better the results. Comparing to the previously used parameter (η≡|∇ ⋅ B|/|∇ × B|), ξ is more relevant for null identification. Using the new method, we reconstruct the magnetic field topology around a radial‐type null and a spiral‐type null, and find that the topologies are well consistent with those predicted in theory. We therefore suggest using this method to find magnetic nulls and reconstruct field topology with four‐point measurements, particularly from Cluster and the forthcoming MMS mission. For the MMS mission, this null‐finding algorithm can be used to trigger its burst‐mode measurements.

Human neutrophil antigens: a nomenclature update based on new alleles and new antigens

In 1998, the ISBT Granulocyte Antigen Working Party established a new nomenclature for the human neutrophil antigens (HNA) which included five antigen systems and a total of seven antigens. In the meantime, new antigens and new alleles have been described and these now make it necessary for the ISBT HNA nomenclature subcommittee to present a proposal for an HNA nomenclature update. HNA‐1 includes four antigens, HNA‐1a, HNA‐1b, HNA‐1c and HNA‐1d, which can result from five different Fcγ receptor IIIb (FcγRIIIb)‐encoding alleles, FCGR3B*01, *02, *03, *04 and *05. Due to copy number variation, individuals may exhibit between zero and four alleles leading to either the HNA‐1 null type or combinations of the known HNA‐1 antigens. Current data indicate that HNA‐2 is an isoantigen without allelic variation, encoded by the CD177 gene. Individuals without any HNA‐2 expression on their neutrophils are addressed as HNA‐2 null. The two antithetical antigens of the choline‐transporter‐like protein 2 (CTL2), HNA‐3a and HNA‐3b, are encoded by the three different alleles SLC44A2*01, *02 and *03, which discriminate in one or two nucleotide substitutions. HNA‐4a, which is encoded by the ITGAM*01 allele, is an antigen of the αMβ2‐integrin. The αLβ2‐integrin contains HNA‐5a as the only antigen which is encoded by the ITGAL*01 allele. Antigens and alleles other than those listed have to be approved by the ISBT Granulocyte Immunobiology Working Party (GIWP) before they can be assigned to the official nomenclature.

110 Missense Mutations of SLC9A3 in Patients With Congenital Sodium Diarrhea Are Associated With Reduced Na+/H+ Exchanger 3 (NHE3) Activity: Identification of the Cause of the Phenotype

Myo5b null mice display microvillus inclusions. Ezrin and p120 immunostaining were assessed in Myo5b null and wild type littermate mice. In a wildtype littermate, ezrin (left panel and red-right panel) was only expressed on the apical surface of the enterocytes. In all Myo5b null mice examined to date, ezrin was found in inclusions within the enterocytes. p120 localization (green-right panel) was unchanged in null mice. Heterozygote mice were identical to wild type mice and did not show microvillus inclusions. Scale bars = 50 μm.