Nonsense Research Articles

Mutations analysis of FRMD7 gene in idiopathic congenital nystagmus families

Objective To reveal the pathogenic mutations in Chinese families with idiopathic congenital nystagmus(ICN) Methods Six families with ICN were recruited from Subei People's Hospital.DNA was extracted from peripheral blood samples of all participants.All coding and exon-intronic boundary regions of the targeted gene FRMD7 were amplified with PCR and sequenced using Sanger sequencing to detect potential pathogenic mutations.This study followed the Helsinki Declaration and was approved by the Ethics Committee of Subei People's Hospital (NO.2015KY-126). All patients or their guardians signed informed consent. Results Three mutations (c.902A>G, c.1944T>A and 1945G>T) were screened in two families after co-segregation validation of intrafamilial genotype-phenotype, c.1944T>A and 1945G>T were newly detected mutations which were not detected in 100 normal controls.No significant mutations were found in the FRMD7 coding region and adjacent splicing sites in the probands of the other four families. Conclusions Two novel pathogenic mutations of FRMD7 are discovered, which expands the pathogenic mutational spectrum of FRMD7 gene causing ICN. Key words: Idiopathic congenital nystagmus; FRMD7 gene; Missense mutation; Nonsense mutation

Imaging nonsense.

Imaging nonsense.

Wyrafinowany koncept. O miejscu [Królewny Lali] Adama Mickiewicza w historii literatury

The article attempts to define the place of Adam Mickiewicz’s unfinished work from the last stage of the poet’s artistic life, titled [Królewna Lala] [Princess Lala], in the history of literature, and thus in the history of literary research, and to outline the possible interpretations of the work using the category of Romantic irony and the idea of concept. A fine artistic idea behind [Królewna Lala] consists in using complex contrasts, combining fairy tale with realism, seriousness with comedy, simplicity of language with poetic mastery, dialogue with literary tradition. It is a virtuoso performance of the writer who creates a comic effect that is closer to the poetics of the absurd (pure nonsense) than to pure comedy. The work reveals a new aesthetic quality, unknown to Mickiewicz’s earlier works.

Reproductive Implications of Consanguinity: Recurrent Hydrops and a Fetal Diagnosis of Lethal Gaucher Disease

Consanguinity is defined as a relationship with one recent ancestor in common. We present a Muslim couple with known consanguinity and their fetal outcomes over multiple pregnancies. This patient is a 28 yo G4P0300 who is married to her fourth cousin. Her first pregnancy was in Egypt during which a fetal demise was diagnosed at 21 weeks. Her second pregnancy ended in another fetal demise at 27 weeks and most organs were consistent with hydrops. Expanded carrier screening of both parents was negative. Her third pregnancy was complicated by hydropic changes at 21 weeks and fetal demise at 22 weeks. She consented to a trio whole exome sequence revealing a homozygous nonsense mutation c.1534A>T in the GBA gene. The mutation was found in the heterozygous state for each parent. Homozygosity for this mutation is associated with the perinatal lethal form of Gaucher disease, type 2. The patient was already 13 weeks pregnant when these results became available and she opted for a CVS procedure which confirmed that the fetus had not inherited any copies of the c.1534A>T mutation and was unaffected. While there is several case reports describing the Gaucher disease type 2, this case highlights the relevance of consanguinity. This case highlights the utility of whole exome sequencing in the setting of prenatal diagnosis and illustrates the significant limitations of expanded carrier screening when a genotyping panel is utilized with limited mutations per gene evaluated.

Clinical features and ion channel gene mutations analysis in 17 cases of early-onset epileptic encephalopathy

Objective To analyze the clinical characteristics and gene mutations of early-onset epileptic encephalopathy(EOEE) caused by ion channel gene mutation, to identify the etiology, to guide the treatment and to provide the basis for genetic counseling. Methods The clinical data from 17 children with EOEE caused by ion channel gene mutation and the peripheral blood of the children and their parents were collected from June 2014 to May 2018 at the Department of Neurology, Tianjin Children′s Hospital.Epilepsy gene sequencing was performed by using disease gene targeting second generation sequencing technology.The mutation of pathogenic ion channel gene was found.The confirmed mutations were verified by Sanger sequencing and the source of the mutation was identified. Results Among 17 case with EOEE, 3 cases had genetic mutation, and 14 cases had denovo mutations.Dravet syndrome was found in 8 cases (47.1%), there were SCN1A gene missense mutation in 5 cases, SCN1A gene nonsense mutation in 3 cases, KCNQ2 gene missense mutation in 1 case (5.9%) and non-specific epileptic encephalopathy in 8 cases (47.1%). SCN2A gene missense mutation, SCN4A gene missense mutation, SCN8A gene missense mutation, KCNQ2 gene missense mutation and KCNH gene missense mutation were found in suspected pathogenic mutations.There were 1 missense mutation out of 5 genes, 1 missense mutation of CACNA1A gene, 1 missense mutation of GRIN2A gene and 1 missense mutation of GRIN3A gene.Seventeen patients were treated with 2 or more antiepileptic drugs, 4 with ketogenic diet and 1 with vitamin B6 supplementation.During 11 to 96 months of follow-up, seizures were completely controlled in 3 cases (17.6%), decreased in 7 cases (41.2%) by more than 50%, and decreased in 7 cases (41.2%) by less than 50%. Conclusions The clinical phenotypes for children with unexplained EOEE are varied, and gene mutations of ion cha-nnel are most common.Some gene sites are denovo mutations which have not been reported such as missense mutation for 3 case SCN1A gene, 1 case SCN2A gene, 1 case CACNA1A gene, 1 case KCNH5 gene, and nonsense mutation for 2 case SCN1A gene, which have enriched the mutation spectrum of EOEE. Key words: Early-onset epileptic encephalopathy; Ion channel; Etiology; Gene mutation

Clinical features of Wiskott-Aldrich syndrome: an analysis of 13 cases

To study the clinical features of Wiskott-Aldrich syndrome (WAS) in children. A retrospective analysis was performed for the clinical data of 13 children with WAS. All 13 children were boys, with a median age of onset of 3 months (range 1-48 months) and a median age of 24 months (range 1-60 months) at the time of diagnosis. Of the 13 children, only 3 had typical WAS and the remaining 10 children had X-linked thrombocytopenia (XLT). The mean WAS score was 2 (range 1-3), the mean platelet count was 20.5×109/L [range (13-46)×109/L], and the mean platelet volume was 8.1 fl (range 6.7-12.1 fl). Lymphocyte subsets and immunoglobulins were measured for 4 children, among whom 1 (25%) had a reduction in both the percentage of CD3+T cells per lymphocyte and lymphocyte per nuclear cells, 1(25%) had a reduction in CD3-CD56+ NK cells. Among these 4 children, 1 (25%) had an increase in IgG, 2 (50%) had a reduction in IgM, 1 (25%) had a reduction in IgA, and 4 (100%) had an increase in IgE. A total of 14 gene mutations belonging to 13 types were found in 13 children, among which there were 9 missense mutations (65%), 2 splicing mutations (14%), 2 nonsense mutation (14%), and 1 frameshift mutation (7%). The median follow-up time was 39 months (range 3-62 months), and all 13 children survived. Children with WAS often have a young age of onset, and most of them are boys. Major clinical features include thrombocytopenia with a reduction in platelet volume. Missense mutation is the main type of gene mutation.

Two cases of hereditary hypomagnesaemia with secondary hypocalcaemia and 4 novel mutations on TRPM6 gene

Objective To investigate the clinical, biochemical and genetic features of hereditary hypomagnesaemia with secondary hypocalcaemia. Methods Two boys came from different Chinese families.They were hospitalized at the Peking University First Hospital between 2014 and 2016 at the age of 9 years and 1 year and 2 months because of epilepsy and psychomotor retardation.Clinical investigation, laboratory examination, and medical imaging were performed for the etiological study.Whole-genome sequencing was used for the genetic analysis of the patients.Mutations of TRPM6 gene were confirmed by means of Sanger sequencing. Results Patient 1 presented with recurrent seizures and psychomotor retardation from the age of 3 months.Vision loss and psychomotor regression were noticed from the age of 9 years, accompanied with hypertension.Serum magnesium and total calcium were significantly decreased to 0.13-0.15 mmol/L and 1.43-2.00 mmol/L, respectively in patient 1.Serum potassium was reduced to 1.85-3.25 mmol/L.Blood parathyroid hormone was also decreased.On the TRPM6 gene of patient 1, 2 novel non-sense mutations, c.2771G>A (p.Trp924Ter) and c. 115C>T (p.Gln39Ter) were identified.Patient 2 presented with seizures and psychomotor retardation at the age of 2 weeks.Both of his serum magnesium (0.17-0.35 mmol/L) and serum total calcium (1.32 - 1.34 mmol/L) were significantly decreased.Blood parathyroid hormone was decreased.Two novel mutations (c.1239G>A, p.W413X and c. 146G>A, p.C49Y) were found in the TRPM6 gene of patient 2.Severe hypomagnesaemia, hypocalcaemia and TRPM6 gene mutations confirmed the diagnosis of hereditary hypomagnesaemia with secondary hypocalcaemia in the 2 patients.After the large-dose supplement of magnesium sulfate, progressive clinical improvements were observed in the 2 patients.However, because of the severe brain damage, patient 1 still had psychomotor retardation.Patient 2 completely recovered. Conclusions Hereditary hypomagnesaemia with secondary hypocalcaemia is a severe inherited metabolic disease.Early diagnosis and large-dose magnesium supplement are the key to the good prognosis of the patients.In this study, 2 Chinese children with the clinical onset of epilepsy and psychomotor retardation are reported.The diagnosis is made by way of blood biochemical assay and gene analysis.Four novel mutations on their TRPM6 gene are identified. Key words: Hypomagnesaemia; Hypocalcaemia; TRPM6 gene; Epilepsy; Inherited metabolic diseases

Mutation spectrum and new mutation of phenylalanine hydroxylase gene in Guangxi region

Objective To investigate the characteristics of the phenylalanine hydroxylase(PAH) gene mutations in patients with phenylketonuria(PKU) in Guangxi region, in order to provide clinical data for genetic counseling and prenatal gene diagnosis. Methods Thirty-seven children diagnosed as PKU in the Maternal and Children′s Hospital of Guangxi Zhuang Autonomous Region were enrolled in the study between January 2009 and December 2017. Venous blood was collected and the PAH gene sequence was determined by Sanger sequencing after amplification with the polymerase chain reaction technique. The new gene mutations were defined based on the national and international literature review and databases.Meanwhile, 100 healthy individuals were selected as the control group for gene sequencing to confirm whether the mutation was a new one. Results Thirty-seven cases of PKU were detected for 68 mutations, with the detection rate being 91.89% (68/74). Six mutations were identified in exon 7, which accounted for 31.08% of all, exon 12(18.92%), exon 8(10.81%) and exon 6 (10.81%) followed.A total of 25 different mutations were identified which including 14 missense mutations(56.00%), 7 nonsense mutations(28.00%), 3 splicing junction mutations(12.00%), and 1 deletion mutation(4.00%). The most common mutations included c. 1223G>A(p.R408Q), c.728G>A(p.R243Q) and c. 721C>T(p.R241C), accounting for 14.86%, 13.51%, and 10.81%, respectively.After querying international databases, including PAH mutation database and Human Gene Mutation Database and forecasting software, three kinds of mutations c. 314C> T (p.T105I), c.583A> G (p.K195E), c .851G> A(p.C284Y) were verified as novel PAH gene mutations. Conclusions The mutation spectrum of the PAH gene in Guangxi has been identified.And 3 kinds of mutations have been identified.This may accumulate valuable information for gene diagnosis and prenatal diagnosis of PKU in Guangxi region. Key words: Phenylketonuria; Phenylalanine hydroxylase; Gene mutation

TRANSFORMATION OF ENGLISH VICTORIAN NONSENSE IN THE 20th CENTURY: LEWIS CARROLL AND JOHN LENNON

TRANSFORMATION OF ENGLISH VICTORIAN NONSENSE IN THE 20th CENTURY: LEWIS CARROLL AND JOHN LENNON

The World Dream As Parable of Being

The bulk of the scientific fraternity still regards the world as ‘a thing as such’, a ‘reality’, in other words, that is independent of an active mind. Its proponents build their theories on facts they regard as something certain, steadfast and unalterable. The etymology of ‘fact’ reveals that its meaning has its roots in the Latin word ‘facere’, ‘to do’. Its present day meaning was established in the 17th century when it meant ‘something that has occurred’. An occurrence is based on observation, which does not enjoy universal consensus, but is an individual matter. Like a dream, which is an occurrence, waking occurrences need interpretation. In light of this a waking fact is no more certain, steadfast and unalterable than a dream fact. Neither of them is real, for the precondition of reality is something unchanging. What, however, is steadfast and unchanging is consciousness. It persists when someone is knocked unconscious and it persists even beyond the death of the body. NDEs provide evidence for this, and so does the ‘death’ of the mystic and yogi. In daily life consciousness may fluctuate, but it will never extinguish. Consciousness is the sine qua non of existence and thus its everlastingness imbues the sense of reality to dreams and waking experience. Absolute Consciousness contains all there is, was and will be. It reveals its unexpanded content by emanating it in form of dreams and waking life where it expands in an illusive show like Plato’s play of shadows or like the cinematic screening or the virtual reality headgear. Like a cinematic screening, existence, which means standing out from something, is a projection. The projector is the brain, which screens the dream world inside the skull and the waking world outside of it. Both projections are illusive. The circumstance that dreams can conjure up vast landscapes within the skull forces us to suspect that the same applies to the waking world. Radin’s innumerable Double Slit experiments that show that consciousness collapses wave functions instantly at enormous distances, together with QM entanglement, makes nonsense of the speed of light and thus of time and space. Consciousness gives up its inner reality in a manner that might be compared to white light of the sun revealing its innate rainbow colours as it passes through a crystal, which latter would be analogous to the brain. NDE subjects often ask if the light they encounter was God. The answer is always ‘no’. This is because that light is the first manifestation of God, or of absolute Consciousness, just as the rays of the sun are its first manifestation and not the sun itself. A reflection of that divine light is ever-present in all life forms. We experience it in NREM Delta sleep. Unfortunately we forget it by the time we wake up since we have passed through the commotion of the last dream.

EFFECT OF LUMOSITY TRAINING SCHEDULE ON RECALL OF NONSENSE SYLLABLES

Randomized between subjects before-after experimental research design was adopted for incontestable establishment of cause-and-effect relationship between Lumosity training schedule and recall of nonsense syllables, if it exists. Learning is mainly a function of memory or recall. A random sample of 150 Tenth Grade students, each aged 15 years, was drawn for the investigation. They were randomized into five groups assigned different schedule of Lumosity Training as experimental conditions (daily 00, 06, 12, 18, and 24 minutes Lumosity training). The experimental treatment lasted for 60 days that was judged long enough for the training to cause some changes in the subjects, if at all Lumosity training could positively or negatively affect the subjects’ memory or recall ability. The 60 brands of Lumosity brain enhancement games used as treatment conditions fall under seven different skills categories (8 on Problem-solving skills, 9 on Flexibility skills, 9 on Speed skills, 11 on Attention skills, 13 on Memory skills, 5 on Math skills, and 5 on Language skills). Results of data analysed with Analysis of covariance (ANCOVA) showed statistically significant effect of Lumosity training schedule on recall of nonsense syllables when the effect of the nonsense syllables Pre-test has been held constant or controlled for [ F (4, 144) = 126.725, p < .05, Partial ɳ 2 = .779] . To ascertain the pairwise mean differences that were significant statistically, Pairwise Multiple Comparisons done revealed significant Mean Difference between each pairwise means in favour of the group that had longer daily Lumosity training. Thus, Lumosity training schedule can be used to overwhelmingly improve recall or memory.

Researches of microRNA-200a on malignant biological behavior of human glioma cells

Objective To investigate the effect of microRNA (miRNA, miR)-200a on proliferation, apoptosis, migration and cell cycle of glioma cell line U251 and its possible mechanism. Methods miR-200a mimics (miR-200a group) and nonsense sequences (Mock group) were transfected into human glioma cell line U251 in vitro by liposome transient transfection. Blank group (Blank group) group). The expression of miR-200a was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) after transfection for 48 hours. The cell proliferation ability of the three groups was detected by cell counting kit-8 (CCK-8) method. The cell migration ability was detected by scratches. Cell apoptosis and cell cycle were detected by flow cytometry The expression of ZEB1 and Cyclin D1 protein was detected by Western blotting. Results Compared with the Mock and Blank groups, the relative expression of miR-200a in miR-200a cells was significantly higher than that in the untreated group (P=0.000, 0.000). The cells of miR-200a cells were significantly different (P=0.028, 0.010). The cell migration ability of miR-200a group was significantly decreased (P=0.031, 0.030), the proportion of G0/G1 phase cells in miR-200a group was increased (P=0.011, 0.013), the proportion of S phase cells was significantly decreased (P=0.024, 0.008), and the apoptosis rate of miR-200a group was significantly lower than that of miR-200a group (P=0.001, 0.001). Western blotting showed that the expression of ZEB1 (P=0.042, 0.034) and the expression of Cyclin D1 were significantly down-regulated (P=0.020, 0.013). Conclusion Upregulation of miR-200a can promote glioma cell apoptosis, block its cell cycle progression and inhibit its migration ability, which is expected to be a new target for the treatment of glioma. Key words: Glioma; MicroRNA-200a; Apoptosis; Cell Cycle; Cell Migration

Wiersze dla niegrzecznych dzieci. Danuta Wawiłow i angielskie nursery rhymes (przekłady, inspiracje)

Among numerous books of poetry for children written by Danuta Wawiłow – famous XXth century Polish poet and writer – one book is special: Poems for Naughty Children [Wiersze dla niegrzecznych dzieci] published in 1987. The book is unique not only because of the poetical qualities it represents – rhythmicity, melodiousness, catchiness, melancholy and lyricism equally with pure nonsense, absurdity, grotesque, (black) humour and numerous equivoques and puns, which are characteristic for Wawiłow’s poetical works; above all else Poems for Naughty Children is a collection of translations of the traditional English nursery rhymes, made by Danuta Wawiłow, who claimed more than once that she was a translator long before she had become a writer. The article aims to explore Wawiłow’s translation work which (up to now) remains almost unknown and un-described in Polish theoretical discourse. Moreover, the author presents the outcome of her research conducted to find “missing originals” of translated nursery rhymes, offers the classification of them and undertakes critical reflection over translation strategy adopted by Danuta Wawiłow.

Mutation analysis of the ADAR1 gene in a pedigree with dyschromatosis symmetrica hereditaria

Objective To detect mutations in the ARAD1 gene in a pedigree with dyschromatosis symmetrica hereditaria (DSH) . Methods Genomic DNA was extracted from the peripheral blood of 8 family members (including 5 patients with DSH and 3 unaffected members) in the pedigree with DSH, as well as 100 unrelated healthy controls. All the 15 exon sequences of the ADAR1 gene were amplified by polymerase chain reaction (PCR) followed by direct sequencing. Then, mutations were detected in comparison with the standard sequence of the ADAR1 gene in Genebank. Results A nonsense mutation C.1420C > T (p.Arg474X) was identified at position 1 420 in exon 2 of the ADAR1 gene in the 5 patients with DSH, but not in the 3 unaffected members or 100 unrelated healthy controls. Conclusion The nonsense mutation C.1420C > T in the ADAR1 gene is the causative mutation in the pedigree with DSH. Key words: Codon, nonsense; Gene, ADAR1; Dyschromatosis symmetrica hereditaria

Diagnosis of red blood cell hexokinase deficiency and literature review

Objective To investigate the value of gene sequencing and protein conformation in the diagnosis process by reporting a case of erythrocyte hexokinase deficiency and reviewing relevant literature. Methods One patient with erythrocyte hexokinase deficiency and his families in the pediatrics of Sun Yat-sen Memorial Hospital of Sun Yat-sen University on May 23, 2016 were selected as the research objects. Clinical data was collected from the initial records by retrospective analysis. Pathogenic gene was detected by target sequence capture and next generation high-throughput sequencing, and the effects of mutant genes on hexokinase function were studied by observing changes in the 3-dimensional structure of proteins. Results The patient exhibited chronic hemolytic anemia after birth. The high-throughput second-generation gene sequencing result of the DNA sample of the child showed that a de novo mutation in the HK1 gene located on chromosome 10, the substitution of cytidine at site 34 of exon1 by thymine : NM_033496: exon1: c. C34T: p. Arg12X, which was a nonsense mutation. Bioinformatics analysis and protein function simulation confirmed that the mutation of the gene can lead to the premature termination of the synthesis of the peptide chain. The short peptide synthesized after the mutation does not contain the binding group and catalytic group of the erythrocyte hexokinase, resulting in the loss of enzyme activity. The patient was eventually diagnosed with erythrocyte hexokinase-deficient hemolytic anemia caused by the HK1 gene mutation. Conclusions In clinical practice, patients with chronic non-spherical erythrocyte hemolytic anemia should be alert to erythrocyte hexosaminidase deficiency disease. Those with conditions should be diagnosed by gene sequencing and protein configuration analysis. Key words: Hexokinase deficiency; Hemolytic anemia; Sequence analysis; Protein structure, quaternary; HK1 gene

Molecular biology study of RHD gene in Chinese Han voluntary blood donors with RhD-negative blood type

Objective To explore the RHD gene structure of Chinese Han voluntary blood donors with RhD-negative blood type, and to conduct a molecular biological analysis of a newly discovered RHD gene mutation. Methods From 2016 to 2017, a total of 85 Chinese Han voluntary blood donors who participated in blood donation in Shandong Blood Center were selected as the subject by randomized digital table. Real-time fluorescence quantitative (RQ)-PCR method was used to detect exon 7 of RHD gene in DNA samples from 85 blood donors. The multiplex PCR-sequence specific primer (SSP) method was used to detect exons 3 to 7 and 9 of RHD gene in DNA samples from these 85 blood donors. The RHD gene in DNA samples from 30 blood donors whose DNA samples expressed RHD gene exon 7 were sequenced using a fluorescently labeled Sanger dideoxy chain termination method. The procedure of this study is in accordance with the requirements of the revised World Medical Association Declaration of Helsinki in 2013. Informed consent was obtained from each blood donor. Results ① In 85 blood donors with RhD-negative blood type, DNA samples of 30 donors had exon 7 of RHD gene, and of 55 blood donors had deletion of exon 7 of RHD gene.② Among the 30 donors with RHD gene exon 7, the DNA samples of 29 donors had exons 3 to 7 and 9 of the RHD gene. Exon 3, 4, 5, 7 and 9 of RHD gene were present in DNA sample of one donor, but exon 6 was deleted. DNA samples from 55 donors without RHD gene exon 7, were deletion of RHD gene exons 3 to 7 and 9. ③ Among the 30 blood donors with RHD gene exon 7, 29 DNA samples had one c. 1227G>A point mutation on RHD gene exon 9. There was a non-sense mutation in c. 933C>A on RHD gene exon 6 in one blood donor′s DNA sample with deletion of the exon 6 of RHD gene. This mutation produced a stop codon, resulting in inconsistent multiplex PCR-SSP and DNA sequencing results of RHD gene in this blood donor. Conclusions The deletion of RHD gene is the main molecular mechanism that causes the RhD-negative phenotype of Chinese Han people. The RHD gene c. 1227G>A mutation is a major feature of Del phenotype. A new c. 933C>A mutation in the RHD gene results in a RhD-negative phenotype. Key words: Rh-Hr blood system; RhD-negativity; Red cells; Immunohematology; Immunogenetics

MYD88 and CXCR4 Mutation Profiling in Lymphoplasmacytic Lymphoma/Waldenstrom's Macroglobulinaemia.

Recurrent mutations affecting MYD88 and CXCR4 gene nowadays form the basis for the diagnosis, risk stratification and use of inhibitors targeting these signalling pathways in LPL/WM which are rare B cell neoplasms. MYD88 L265P mutation analysis was performed on 33 cases of LPL/WM by AS-PCR (positivity-84.8%, n = 28/33) and by Sanger sequencing (positivity-39.3%, n = 13/33). We had only two cases with CXCR4 non-sense (NS) mutation (p.S338*) using Sanger sequencing. MYD88 (L265P) mutation detection by AS-PCR can form reliable biomarker for the diagnosis of LPL/WM in molecular labs. Although the cohort is small, still the CXCR4 mutation frequency in our study is low as compared to the published literature.

Abstract 1814: NF1 as an estrogen receptor-α co-repressor in breast cancer

Abstract NF1 has been best known as a GAP (GTPase Activating Protein) that inactivates Ras. However, we are now finding evidence that it also functions as an ER co-repressor, whose loss leads to endocrine therapy resistance. Sequencing tumor DNA from &amp;gt;600 ER+ breast cancers treated by tamoxifen adjuvant monotherapy, we found that frameshift (FS) and nonsense (NS) NF1 mutations, which can create an NF1-null state, strongly correlate with relapse risk (HR=2.6, submitted). Surprisingly, no recurrent missense NF1 mutations inactivating GAP activity were found in our cohort, and such mutations are rare in primary cancers in general. We thus posulated that complete loss of NF1 protein (e.g., caused by NS/FS mutations), but not GAP inactivation alone, is required to drive endocrine therapy resistance. Here we demonstrate that NF1 loss (by gene silencing) in ER+ breast cancer cells greatly enhances ligand-dependent ER transcriptional activity in vitro and in vivo, causing estradiol (E2) hypersensitivity and tamoxifen agonism. Mechanistically we show that NF1 can bind directly to ER, an interaction enhanced by tamoxifen but not by E2. Binding is mediated by leucine/isoleucine-rich motifs in NF1, analogous to other ER co-repressors. Mutations in these motifs (some of which are targeted by somatic mutation in cancer) inhibit ER binding and transcriptional activity without impacting GAP activity; conversely, inactivating GAP activity does not impact ER binding and repression. To validate NF1 as an ER co-repressor, we examined proteomic data from &amp;gt;100 breast cancer patients in the CPTAC data base and found that proteins whose levels are positively correlated with NF1 are highly enriched with factors known to bind nuclear receptors; by contrast, levels of another GAP, p120, which lacks ER binding sites, are negatively correlated with these molecules. Importantly, preclinical treatment studies indicate that while NF1-deficient ER+ breast cancer should not be treated by tamoxifen or aromatase inhibitors, fulvestrant, which degrades ER, remains effective. However, fulvestrant monotherapy can activate the Ras-MAP pathway, which may promote cell survival and acquired fulvestrant resistance unless combined with dabrafinib and trametinib to inhibit Raf and MEK —a clinical trial for this combination is in development. Our data suggest that NF1 is a dual negative regulator at the intersection of two potent oncogenic signaling pathways, Ras and ER. Combination therapy targeting both the ER and the Ras-Raf pathways should be investigated for NF1-deficient cancers driven by ER. Citation Format: Eric C. Chang, zeyi Zheng, Meenakshi Anurag, Jin Gao, Burcu Cakar, Xinhui Du, Jing Li, Philip Lavere, Jonathan T. Lei, Purba Singh, Sinem Seker, Wei Song, Jianheng Peng, Tiffany Nguyen, Doug Chan, Xi Chen, Kimberly C. Banks, Richarad B. Lanman, Maryam Shafaee, Susan Hilsenbeck, Charles Foulds, Matthew J. Ellis. NF1 as an estrogen receptor-α co-repressor in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1814.

Roles of Dectin-1 in phagocytosis of Candida albicans by macrophage-like cells derived from a human acute monocytic leukemia cell line THP-1

Objective To investigate the roles of Dectin-1 in phagocytosis of Candida albicans (C. albicans) by macrophage-like cells derived from a human acute monocytic leukemia cell line THP-1. Methods THP-1 macrophage-like cells served as the target cells, and were transfected with small interfering RNA (siRNA) targeting Dectin-1 to down-regulate the expression of Dectin-1 receptor (siRNA-Dectin-1 group) . THP-1 macrophage-like cells transfected with nonsense siRNA (siRNA-NC) served as a negative control group. After transfection, the THP-1 macrophage-like cells in the above 2 groups were co-cultured with heat-killed C. albicans separately. And then, fluorescence microscopy was performed to count THP-1 macrophage-like cells phagocytosing C. albicans, and flow cytometry was used to determine the mean fluorescence intensity (MFI) of dihydrorhodamine (DHR) -123 fluorescent cells. Statistical analysis was done by one-way analysis of variance (ANOVA) and t test with the SPSS19.0 software. Results After transfection with siRNA-Dectin-1, the mRNA and protein expression of Dectin-1 significantly decreased in THP-1 macrophage-like cells (t= 26.163, P < 0.001) . After 1-, 2-, 4-hour co-culture of THP-1 macrophage-like cells with C. albicans, fluorescence microscopy showed that the phagocytosis rates of C. albicans by THP-1 macrophage-like cells were significantly lower in the siRNA-Dectin-1 group than in the negative control group (17.5% vs. 22.1%, 18.6% vs. 24.3%, 39.2% vs. 59.1%, respectively, all P < 0.05) , so were the percentage of THP-1 macrophage-like cells phagocytosing more than 3 C. albicans cells (2.2% vs. 4.7%, 2.5% vs. 5.4%, 5.1% vs. 8.3%, respectively, all P < 0.05) . After 30-minute, 1-, 2- and 4-hour co-culture of THP-1 macrophage-like cells with DHR-123-labelled C. albicans, flow cytometry showed that the MFI of C. albicans-phagocytosing cells was significantly lower in the siRNA-Dectin-1 group than in the negative control group (36.8 vs. 45.7, 54.3 vs. 62.4, 72.1 vs. 84.9, 93.6 vs. 116.7, respectively, all P < 0.05) . Conclusion Dectin-1 receptor plays an important role in the phagocytosis of C. albicans by macrophages. Key words: Candida albicans; Macrophages; Phagocytosis; Leukemia, monocytic, acute; Dectin-1

The development of the Humor Structure Appreciation Scale and its relation to Sensation Seeking Inventory and Need for Closure Scale

Three related studies are presented with the aim of investigating the individual differences in humour appreciation, sensation seeking and need for closure. In order to accomplish this, the construction and initial validation of a new instrument will be presented. The Humour Structure Appreciation Scale (HSAS) distinguishes between incongruity-resolution (INC-RES), and nonsense (NON) humour. The first study analyses the content validity of a broad item pool. The second study focused on the metric properties of the selected items. In the third study, the remaining jokes were given to a quota sample from Macedonia, along with measures of sensation seeking (Arnett Inventory of Sensation Seeking) and need for closure (Need for Closure Scale). Psychometric properties of the scales were investigated and showed to be satisfactory. Intercorrelations among the scales provide evidence for the construct validity of the HSAS.