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Open in new tab Download slide OBJECTIVES The Bex-Nikaidoh operation can effectively relieve left ventricular outflow tract obstruction. However, if a conduit is used for right ventricular outflow tract reconstruction, a late reoperation can be anticipated. We examined the impact of double root translocation on outcomes. METHODS We performed a retrospective single-centre study of patients who underwent aortic root translocation between 2006 and 2019. RESULTS Aortic root translocation was performed in 23 patients at a median age of 1.6 years [interquartile range (IQR) 0.9–2.5]. Concomitant repairs were done in 52.2% of patients (12/23) including the Senning atrial switch in 34.8% (8/23). The right ventricular outflow tract was reconstructed with valved conduits in 39.1% (9/23), direct anastomoses in 4.35% (1/23) and pulmonary autografts in 56.5% of patients (13/23). Aortic cross-clamp time was significantly longer in patients with double root translocation [308 min (IQR 270–259) vs 209 min (IQR 179–281), P  =   0.02]; 2 patients in this group required temporary mechanical circulatory support. There were no early deaths. Median follow-up time was 7.5 years (IQR 3.3–10.5). The estimated 10-year survival was 90% [95% confidence interval (CI): 47.3%, 98.5%]. There was no recurrent left ventricular outflow tract obstruction. Freedom from any reoperation was 64.2% (95% CI: 40.8%, 80.3%) at 3 years and 44.5% (95% CI: 21.2%, 65.5%) at 6 years. The main indication for late reoperation was conduit degeneration. Freedom from a right ventricular outflow tract reoperation was significantly higher, and the number of reoperations per patient was lower when a double root translocation had been performed (P  =   0.03). CONCLUSIONS The Bex-Nikaidoh operation effectively relieved left ventricular outflow tract obstruction. A double root translocation further increased procedural complexity but was associated with better mid-term freedom from a right ventricular outflow tract reoperation. It should be considered in suitable patients. [ABSTRACT FROM AUTHOR]

Objectives Data regarding bone marrow (BM) sampling and cytogenetic testing rates for identification of translocation (11q13;14q32) and their changes over time in a multiple myeloma (MM) population are limited. We analyzed these metrics at a clinic specializing in the treatment of MM. Methods A total of 760 BM aspirate samples from 351 patients were collected between August 2004 and October 2021. We analyzed BM sampling statistics, cytogenetic testing frequency, and the incidence rates for the t(11;14) translocation in a single clinic specializing in the treatment of MM. Results We report that most (54.4%) patients had only 1 aspirate collected; the main reason (64.6%) for BM collection was to confirm disease progression. Less than half (47.5%) of BM samples collected for evaluation of MM disease had cytogenetic testing, but the rates have markedly increased in recent years. Our data demonstrated an incidence rate of 19.3% for t(11;14). Conclusions This report suggests that some patients may need to retest for this genetic aberration due to the possibility of false negatives and the potential benefit of identifying the t(11;14) marker for patients who may be candidates for a highly effective targeted therapy consisting of the BCL-2 inhibitor venetoclax. [ABSTRACT FROM AUTHOR]

Objectives Fluorescence in situ hybridization (FISH) assays for the detection of chromosomal rearrangements involving TFE3 and TFEB are considered the gold standard for the diagnosis of MiTF family altered renal cell carcinoma (MiTF-RCC). We reviewed 801 clinical TFE3 / TFEB FISH assays performed at our tertiary-level institution between 2014 and 2023 on kidney tumors suspicious at the morphologic or biomarker level for MiTF aberrations. Methods We summarized and analyzed clinical information, TFE3 / TFEB FISH results, and available biomarker staining results in a cohort of 453 consecutive kidney tumor cases suspicious for MiTF-RCC. Results In total, 61 of 434 (14%) kidney tumors were confirmed for TFE3 translocation; 10 of 367 cases (2.7%) were confirmed for TFEB translocation. Since TFEB amplification interpretation was implemented in our service line, 20 of 306 cases (6.5%) were diagnosed with TFEB amplification. Importantly, TFE3 and TFEB rearrangements were never co-detected within the same kidney tumor. Patients with TFEB amplification were significantly older (P <.001) than patients with TFE3 or TFEB translocation. Kidney tumors with TFEB amplification were seen to be at least 3 times as common as those with TFEB translocation. Conclusions Clinical TFE3 / TFEB FISH assays successfully identified and confirmed rare MiTF-RCC with TFE3 and TFEB rearrangements. Although morphologic and biomarker features associated with a kidney tumor may be suggestive of MiTF-RCC, clinical TFE3 / TFEB FISH assays are crucial for a confirmation and definitive subclassification of patients with MiTF-RCC. [ABSTRACT FROM AUTHOR]

Background: Uropathogenic Escherichia coli (UPEC) are a major cause of urinary tract infection (UTI), one of the most common infectious diseases in humans. UPEC are increasingly associated with resistance to multiple antibiotics. This includes resistance to third-generation cephalosporins, a common class of antibiotics frequently used to treat UTI.Methods: We employed a high-throughput genome-wide screen using saturated transposon mutagenesis and transposon directed insertion-site sequencing (TraDIS) together with phenotypic resistance assessment to identify key genes required for survival of the MDR UPEC ST131 strain EC958 in the presence of the third-generation cephalosporin cefotaxime.Results: We showed that blaCMY-23 is the major ESBL gene in EC958 responsible for mediating resistance to cefotaxime. Our screen also revealed that mutation of genes involved in cell division and the twin-arginine translocation pathway sensitized EC958 to cefotaxime. The role of these cell-division and protein-secretion genes in cefotaxime resistance was confirmed through the construction of mutants and phenotypic testing. Mutation of these genes also sensitized EC958 to other cephalosporins.Conclusions: This work provides an exemplar for the application of TraDIS to define molecular mechanisms of resistance to antibiotics. The identification of mutants that sensitize UPEC to cefotaxime, despite the presence of a cephalosporinase, provides a framework for the development of new approaches to treat infections caused by MDR pathogens. [ABSTRACT FROM AUTHOR]

Accumulating data suggest that metadherin (MTDH) may function as an oncogene. Our previous study showed that MTDH promotes hepatocellular carcinoma (HCC) metastasis via the epithelial-mesenchymal transition. In this study, we aim to further elucidate how MTDH promotes HCC metastasis. Using Co-immunoprecipitation (co-IP) and mass spectrometry, we found that MTDH can specifically bind to protein arginine methyltransferase 5 (PRMT5). Further functional assays revealed that PRMT5 overexpression promoted the proliferation and motility of HCC cells and that knockout of PRMT5 impeded the effect of MTDH. The immunohistochemistry assay/tissue microarray results showed that when MTDH was overexpressed in HCC cells, PRMT5 translocated from the nucleus to the cytoplasm, with the subsequent translocation of β-catenin from the cytoplasm to the nucleus and upregulation of the WNT–β-catenin signaling pathway. Further in vivo experiments suggested that PRMT5 and β-catenin played a pivotal role in MTDH-mediated HCC metastasis. We therefore concluded that the MTDH–PRMT5 complex promotes HCC metastasis by regulating the WNT–β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Epithelial ovarian cancer (EOC) has a very poor prognosis because of tumor invasiveness. Here, we reported that SET and MYND domain containing protein 3 (SMYD3), a lysine methyltransferase, was frequently upregulated in EOC and associated with poor prognosis. A series of in vitro assays demonstrated that SMYD3 significantly upgraded the migration ability of EOC cells. The results of in vivo EOC metastasis models further confirmed that overexpression of SMYD3 promoted EOC progression. Mechanistic investigations indicated that SMYD3 cloud decrease p53 protein stability and induce epithelial–mesenchymal transition in EOC cells. SMYD3 interacts with p53 directly via the post-SET domain and destabilizes p53 by inducing p53 translocation from the nucleus to the cytoplasm and promoting p53 ubiquitination modification independent of MDM2. Furthermore, the mass spectrometry results showed that SMYD3 interacts with UBE2R2, an ubiquitin-conjugating enzyme (E2) of the ubiquitin–proteasome pathway. The combination of UBE2R2–SMYD3–p53 significantly promotes the ubiquitination and degradation of p53. These results pointed that SMYD3 might be a new E3 ligase of p53. Further analysis confirmed that lysines 381, 382 and 386 of p53 are the key sites for the ubiquitination modification of SMYD3 to p53. In summary, our results define the important role of SMYD3 in the metastasis process of EOC and present a new therapeutic target against EOC. [ABSTRACT FROM AUTHOR]

Background: Adult acquired buried penis syndrome may be associated with an inability to void, sexual dysfunction, and recurrent infection. Previously published classification systems rely on intraoperative findings, such as penile skin quality.Objectives: The purpose of this study was to evaluate outcomes after adult acquired buried penis repair and to develop a classification system based on preoperative assessment.Methods: The authors reviewed data from patients who underwent buried penis reconstruction at a single institution. Patient history and physical examination guided the development of a classification system for surgical planning.Results: Of the 27 patients included, the mean age was 56 ± 15 years and mean body mass index was 49 ± 14 kg/m2. Patients were classified into 4 groups based on examination findings: (I) buried penis due to skin deficiency, iatrogenic scarring, and/or diseased penile skin (n = 3); (II) excess abdominal skin and fat (n = 6); (III) excess skin and fat with diseased penile skin (n = 16); and (IV) type III plus severe scrotal edema (n = 2). Surgical treatment (eg, excision and grafting, mons suspension, panniculectomy, translocation of testes, and/or scrotectomy) was tailored based on classification. Complications included wound breakdown (n = 3), cellulitis (n = 4), and hematoma (n = 1). Nearly all patients (96%) reported early satisfaction and improvement in their symptoms postoperatively.Conclusions: Classifying patients with buried penis according to preoperative examination findings may guide surgical decision-making and preoperative counseling and allow for optimized aesthetics to enhance self-esteem and sexual well-being.Level Of Evidence:4: [ABSTRACT FROM AUTHOR]

Study Question: Can reduced representation genome sequencing offer an alternative to single nucleotide polymorphism (SNP) arrays as a generic and genome-wide approach for comprehensive preimplantation genetic testing for monogenic disorders (PGT-M), aneuploidy (PGT-A) and structural rearrangements (PGT-SR) in human embryo biopsy samples?Summary Answer: Reduced representation genome sequencing, with OnePGT, offers a generic, next-generation sequencing-based approach for automated haplotyping and copy-number assessment, both combined or independently, in human single blastomere and trophectoderm samples.What Is Known Already: Genome-wide haplotyping strategies, such as karyomapping and haplarithmisis, have paved the way for comprehensive PGT, i.e. leveraging PGT-M, PGT-A and PGT-SR in a single workflow. These methods are based upon SNP array technology.Study Design, Size, Duration: This multi-centre verification study evaluated the concordance of PGT results for a total of 225 embryos, including 189 originally tested for a monogenic disorder and 36 tested for a translocation. Concordance for whole chromosome aneuploidies was also evaluated where whole genome copy-number reference data were available. Data analysts were kept blind to the results from the reference PGT method.Participants/materials, Setting, Methods: Leftover blastomere/trophectoderm whole genome amplified (WGA) material was used, or secondary trophectoderm biopsies were WGA. A reduced representation library from WGA DNA together with bulk DNA from phasing references was processed across two study sites with the Agilent OnePGT solution. Libraries were sequenced on an Illumina NextSeq500 system, and data were analysed with Agilent Alissa OnePGT software. The embedded PGT-M pipeline utilises the principles of haplarithmisis to deduce haplotype inheritance whereas both the PGT-A and PGT-SR pipelines are based upon read-count analysis in order to evaluate embryonic ploidy. Concordance analysis was performed for both analysis strategies against the reference PGT method.Main Results and the Role Of Chance: PGT-M analysis was performed on 189 samples. For nine samples, the data quality was too poor to analyse further, and for 20 samples, no result could be obtained mainly due to biological limitations of the haplotyping approach, such as co-localisation of meiotic crossover events and nullisomy for the chromosome of interest. For the remaining 160 samples, 100% concordance was obtained between OnePGT and the reference PGT-M method. Equally for PGT-SR, 100% concordance for all 36 embryos tested was demonstrated. Moreover, with embryos originally analysed for PGT-M or PGT-SR for which genome-wide copy-number reference data were available, 100% concordance was shown for whole chromosome copy-number calls (PGT-A).Limitations, Reasons For Caution: Inherent to haplotyping methodologies, processing of additional family members is still required. Biological limitations caused inconclusive results in 10% of cases.Wider Implications Of the Findings: Employment of OnePGT for PGT-M, PGT-SR, PGT-A or combined as comprehensive PGT offers a scalable platform, which is inherently generic and thereby, eliminates the need for family-specific design and optimisation. It can be considered as both an improvement and complement to the current methodologies for PGT.Study Funding/competing Interest(s): Agilent Technologies, the KU Leuven (C1/018 to J.R.V. and T.V.) and the Horizon 2020 WIDENLIFE (692065 to J.R.V. and T.V). H.M. is supported by the Research Foundation Flanders (FWO, 11A7119N). M.Z.E, J.R.V. and T.V. are co-inventors on patent applications: ZL910050-PCT/EP2011/060211- WO/2011/157846 'Methods for haplotyping single cells' and ZL913096-PCT/EP2014/068315 'Haplotyping and copy-number typing using polymorphic variant allelic frequencies'. T.V. and J.R.V. are co-inventors on patent application: ZL912076-PCT/EP2013/070858 'High-throughput genotyping by sequencing'. Haplarithmisis ('Haplotyping and copy-number typing using polymorphic variant allelic frequencies') has been licensed to Agilent Technologies. The following patents are pending for OnePGT: US2016275239, AU2014345516, CA2928013, CN105874081, EP3066213 and WO2015067796. OnePGT is a registered trademark. D.L., J.T. and R.L.R. report personal fees during the conduct of the study and outside the submitted work from Agilent Technologies. S.H. and K.O.F. report personal fees and other during the conduct of the study and outside the submitted work from Agilent Technologies. J.A. reports personal fees and other during the conduct of the study from Agilent Technologies and personal fees from Agilent Technologies and UZ Leuven outside the submitted work. B.D. reports grants from IWT/VLAIO, personal fees during the conduct of the study from Agilent Technologies and personal fees and other outside the submitted work from Agilent Technologies. In addition, B.D. has a patent 20160275239 - Genetic Analysis Method pending. The remaining authors have no conflicts of interest. [ABSTRACT FROM AUTHOR]

BACKGROUND. The disrupted in schizophrenia 1 (DISCI) gene locus was originally identified in a Scottish pedigree with a high incidence of psychiatric disorders that is associated with a balanced t(1;11)(q42.1;q14.3) chromosomal translocation. Here, we investigated whether members of this family carrying the t(1;11)(q42.1;q14.3) translocation have a common brainrelated phenotype and whether this phenotype is similar to that observed in schizophrenia (SCZ), using multivariate pattern recognition techniques. METHODS. We measured cortical thickness, cortical surface area, subcortical volumes, and regional cerebral blood flow (rCBF) in healthy controls (HC) (n = 24), patients diagnosed with SCZ (n = 24), patients diagnosed with bipolar disorder (BP) (n = 19), and members of the original Scottish family (n = 30) who were either carriers (T+) or noncarriers (T-) of the DISCI translocation. Binary classification models were developed to assess the differences and similarities across groups. RESULTS. Based on cortical thickness, 72% of the T- group were assigned to the HC group, 83% of the T+ group were assigned to the SCZ group, and 45% of the BP group were classified as belonging to the SCZ group, suggesting high specificity of this measurement in predicting brain-related phenotypes. Shared brain-related phenotypes between SCZ and T+ individuals were found for cortical thickness only. Finally, a classification accuracy of 73% was achieved when directly comparing the pattern of cortical thickness of T+ and T- individuals. CONCLUSION. Together, the results of this study suggest that the DISCI translocation may increase the risk of psychiatric disorders in this pedigree by affecting neurostructural phenotypes such as cortical thickness. FUNDING. This work was supported by the National Health Service Research Scotland, the Scottish Translational Medicine Research Collaboration, the Innovative Medicines Initiative (IMI), the Engineering and Physical Sciences Research Council (EPSRC), The Wellcome Trust, the National Institute of Health Research (NIHR), and Pfizer., Introduction Schizophrenia (SCZ) can be a chronic, debilitating psychiatric disorder; it affects approximately 1% of the adult population (1). Neuroimaging has consistently demonstrated widespread structural and functional brain abnormalities (2-5). [...]

Aryl chloroethyl ureas (CEUs) are new protein alkylating agents exhibiting anticancer activity both in vitro and in vivo. We report herein that [sup.14]C-labeled CEU derivatives, designated CEU-025 and CEU-027, covalently bind to thioredoxin-1 (TRX1). Covalent binding of these molecules slightly decreases the disulfide-reducing activity of recombinant TRX1, when compared with the effect of strong thioalkylating agents such as N-ethylmaleimide. Moreover, site-directed mutagenesis and diamide competition assays demonstrated that TRX1 cysteinyl residues are not the prime targets of CEUs. CEU-025 abrogates the nuclear translocation of TRX1 in human cancer cells. In addition, we show that CEU-025 can block TRX1 nuclear translocation induced by cisplatin. Unexpectedly, pretreatment with sublethal CEU-025 concentrations that block TRX1 nuclear translocation protected the cells against cisplatin cytotoxicity. Overexpression of TRX1 in HT1080 fibrosarcoma cells attenuated CEU-025 cytotoxicity, while its suppression using TRX1-specific siRNA increased the effects of CEU-025, suggesting that loss of function of TRX1 is involved, at least in part, in the cytotoxic activity of CEU-025. These results suggest that CEU-025 and CEU-027 exhibit anticancer activity through a novel, unique mechanism of action. The importance of TRX1 and the dependence of the cytotoxicity of CEU-025 and CEU-027 on TRX1 intracellular localization are also discussed. Key words: alkylating agents, thioredoxins, redox signaling, cancer, nuclear translocation, apoptosis, cell proliferation. Les aryl chloroethyl urees (CEU) sont de nouveaux agents alkylants presentant une activite anticanceereuse in vitro et in vivo. Dans le present article, nous indiquons que les derives des CEU marques au [sup.14]C, CEU-025 et CEU-027, se lient de maniere covalente a la thioredoxine-l (TRX1). La liaison covalente de ces molecules diminue legerement l'activite de reduction des ponts disulfures de la TRX1 recombinante, par comparaison avec l'effet d'agents thio-alkylants puissants comme le N-ethymaleimide. Nous demontrons aussi par mutagenese dirigee et des essais de competition avec le diamide que les residus cysteinyl de la TRX1 ne sont pas les principales cibles des CEU. CEU-025 supprime la translocation nucleaire de la TRX1 dans les cellules canceereuses humaines. De plus, nous montrons que CEU-025 peut bloquer la translocation nucleaire de TRX1 induite par le cisplatine. Contre toute attente, un pretraitement avec des concentrations subletales de CEU-025 bloquant la translocation nucteaire de TRX1 a protege les cellules contre la cytotoxicite du cisplatine. La surexpression de TRX1 dans les fibrosarcomes HT1080 a attenue la cytotoxicite de CEU-025, alors que l'inhibition de son expression par siARN a augmente les effets de l'agent, ce qui laisse croire que la perte de fonction de la TRX1 est impliqueee, du moins en partie, dans l'activite cytotoxique de CEU-025. Ces resultats donnent a penser que CEU-025 et CEU-027 ont une activite anticancereuse par l'intermeediaire d'un nouveau et unique meecanisme d'action. Nous examinons aussi l'importance de TRX1 et la dependance de la toxicite de CEU-025 et de CEU-027 envers la localisation intracellulaire de TRXl. Mots-cles: agents alkylants, thioreedoxine, signalisation redox, cancer, translocation nucleeaire, apoptose, proliferation cellulaire. [Traduit par la Redaction], Introduction Aryl chloroethyl ureas (CEUs) are 'soft' alkylating agents that inhibit cell proliferation in vitro and in vivo. CEUs were designed by molecular hybridization of 2 families of 'strong' alkylating [...]

Objective: We re-evaluated reported associations between tobacco use and other factors and non-Hodgkin lymphoma (NHL) t(14; 18)-subtypes based on fluorescence in situ hybridization (FISH) assays believed to be more sensitive than polymerase chain reaction (PCR), previously used for detecting t(14; 18).Methods: Commercial FISH assays and bcl-2 immunostaining were performed on paraffin sections to determine t(14; 18) and bcl-2 case-subtypes. Polytomous logistic regression models estimated associations between NHL case-subtypes (versus 1,245 population-based controls) and tobacco use as well as other factors.Results: Adjusting for age, state, and proxy status, t(14; 18)-negative NHL was associated with any tobacco use (vs. no tobacco use, OR = 1.9, 95% CI = 1.0-3.5), including current smoking (vs. no cigarette use, OR = 1.9, 95% CI = 1.1-3.2). Tobacco exposures were not clearly associated with t(14; 18)-positive NHL or bcl-2 case-subtypes. Hair-dye use and family history of a hemolymphatic cancer were associated with t(14; 18)-negative NHL, but the number of exposed cases was small.Conclusions: The association between t(14; 18)-negative NHL and cigarette smoking was unexpected given previous evidence of associations between smoking and follicular lymphoma (which is largely t(14; 18)-positive). Future studies characterizing additional molecular characteristics of t(14; 18)-negative NHL may help determine whether the association with smoking may have been causal versus an artifact of chance or bias. [ABSTRACT FROM AUTHOR]

The Disrupted in Schizophrenia (DISC) locus on human chromosome 1q42 has been strongly implicated by genetic studies as a susceptibility locus for major mental illnesses. In humans the locus is transcriptionally complex, with multiple alternate splicing events, antisense transcription, and intergenic splicing all evident. We have compared the genomic sequence and transcription maps of this locus between human, mouse, pufferfish (Fugu rubripes), and, in part, zebrafish (Danio rerio). The order and orientation of EGLN1, TSNAX, and DISC1 genes are conserved between mammals and F. rubripes. Intergenic splicing and short intergenic transcripts are not found to be conserved features. DISC2, a putative noncoding transcript partially antisense to DISC1, is not conserved in mouse or F. rubripes. Alternate splice forms of the protein-coding DISC1 gene are conserved even though the genomic structure is not. The amino acid sequence of DISC1 is diverging rapidly, although a putative nuclear localization signal and discrete blocks of coiled coil are specifically conserved features. [Copyright &y& Elsevier]

Chromosome banding analysis was performed on five cases of human malignant melanoma. Results demonstrated a variety of chromosome alterations, including deletion or translocation of chromosome 6q, in four of five cases. A comparison of our results with previously published studies suggests that chromosome 6q may represent a specific site of chromosome aberration in malignant melanoma.

The recognition of recurring sites of chromosome change in human cancers has pinpointed the location in the genome of several important growth-regulatory sequences (e.g., cellular oncogenes). This report details the finding of a recurring translocation site involving the long arm of chromosome 6 (6q) in malignant melanoma. We have observed a translocation (t) between chromosomes 1 and 6 in five different cases of malignant metastatic melanoma. All five melanomas evidencing t(1;6) involved band regions 6q11-13, while two different regions of chromosome 1 (p22, q12-q21) were shown to be translocated to 6q. In reviewing previously published cases of melanoma, an additional two cases of t(1;6) and 13 cases of other translocations to 6q11-13 have been identified. Chromosome 6q contains several biologically important gene sequences including the proto-oncogenes ros, myb, and mas1. However, based on current mapping studies, the breakpoint of this translocation (6q11-13) is not within the region encoding these sequences. By analogy to other systems, molecular analysis of the translocation breakpoints may identify a gene(s) which plays a role in melanoma tumorigenesis.

Complex cytogenetic abnormalities have been described in patients with multiple myeloma (MM). To better understand the significance of the most frequent translocation observed in MM, we studied the clinical characteristics of patients with MM and the t(11;14)(q13;q32) abnormality. A search of the cytogenetic database at the Mayo Clinic identified patients with MM and t(11;14)(q13;q32). The medical records were reviewed for the clinical characteristics of these patients. We identified 13 patients with MM and t(11;14)(q13;q32) determined by standard cytogenetic analysis; in 10 patients the abnormality was detected at the time of relapse (three with previously normal results of cytogenetic examination). At the time the translocation was detected, plasma cell (PC) leukaemia was clinically diagnosed in two patients. The median number of circulating PCs, as determined by the cytoplasmic immunofluorescence of T-cell-depleted peripheral blood mononuclear cells, was 1.1 × 109/l (mean 1.74; range 0.0017–6.26 × 109/l). On linear regression analysis there was a strong correlation between the number of circulating PCs and the number of bone marrow PCs. The median survival after demonstration of the translocation was 8.1 months. Of all patients, 10 died of disease progression and three were alive. Patients with MM who have t(11;14)(q13;q32) seem to have an aggressive clinical course, even when the abnormality is detected at the time of diagnosis, with evidence of many circulating PCs. [ABSTRACT FROM AUTHOR]

The t(9;11)(p21–22;q23) translocation is frequently associated with acute monoblastic leukemia but may occasionally be seen in patients with acute lymphoblastic leukemia (ALL). We report a case of childhood ALL associated with t(9;11)(p21–22;q23) as the unique recurring chromosomal abnormality. A 3-month-old girl presented with “lymphomatous” ALL (renal enlargement), a high leukocyte count and central nervous system (CNS) involvement. Leukemic cell typing revealed a sIg+ B-cell immunophenotype without CD10 and CD34 antigenic expression while the blast cell morphology was of the FAB-L1 type. Splitting of a YAC encompassing the MLL gene was shown by fluorescence in situ hybridization (FISH) studies of the patient’s metaphase chromosomes. Rearrangement of the MLL gene was confirmed by Southern blot analysis. Despite treatment with an hyperintensive polychemotherapeutic regimen, the patient achieved a complete remission but relapsed 9 months later. These results provide further evidence that the t(9;11) may be observed in ALL, involves the MLL gene and is associated with a poor outcome. Moreover, this observation clearly illustrates that sIg+ B-cell ALL is not necessarily associated with a Burkitt (L3) morphology. [ABSTRACT FROM AUTHOR]

Background Healthcare-associated bloodstream infections (HABSIs) are a significant cause of mortality and morbidity in the neonatal intensive care unit (NICU) population. Our objectives were to review the epidemiology of HABSIs in our NICU and to examine the applicability of National Healthcare Safety Network (NHSN) definitions to the NICU population. Methods We performed a retrospective review of all neonates admitted to the 54-bed level IV NICU at Yale-New Haven Children's Hospital with a HABSI between January 1, 2013 and December 31, 2018. HABSI was defined as a positive blood culture at >72 hours of life growing an organism not considered a contaminant. Clinical definitions per treating NICU team and NHSN site-specific definitions were compared for source attribution using McNemar's Chi-square test. Results We identified 88 HABSIs with an incidence rate of 0.81 per 1,000 patient-days. Only 13% of these were central line-associated bloodstream infections (CLABSIs). Infants with a HABSI had median birth weight and gestational age of 830 grams and 26 weeks, respectively, with a high percentage requiring mechanical ventilation parenteral nutrition and vascular access (Table 1). Sepsis-related mortality was 24%. The majority of HABSIs were caused by gram-positive and gram-negative bacteria (Figure 1). Most were secondary to necrotizing enterocolitis, pneumonia or a source that was not identified (Table 2). NHSN definitions were less likely to identify a source compared with clinical definitions per NICU treating team (P < 0.001, Table 2). Fifty percent of patients without an identified source of infection by NHSN criteria were identified with a Mucosal Barrier Injury (MBI) organism, likely causing bacteremia from gut translocation. Conclusion HABSIs occur in premature babies with comorbidities, and are more prevalent than CLABSIs. Gut translocation with MBI organisms may be an important unidentified source of HABSIs in neonates. With the increasing focus on HABSI prevention, there is a need for better NHSN definitions for source attribution of bloodstream infections in neonates. Disclosures All authors: No reported disclosures. [ABSTRACT FROM AUTHOR]

Background Strongyloides stercoralis often causes an asymptomatic infection despite continuous autoinfection for the lifetime of the host. Both HIV and recurrent enteric parasitic infections cause gut damage and increased microbial translocation, but little is known about the effects of co-infection. We aimed to evaluate changes in immune activation, mucosal damage, and microbial translocation in people with HIV-1 (PWH) and parasite co-infection. Methods In this pilot prospective cohort study, we enrolled foreign-born PWH on suppressive antiretroviral therapy (ART) in an ambulatory clinic in Houston, Texas. We evaluated serum Strongyloides IgG using ELISA with an S. stercoralis -specific recombinant protein. Intestinal fatty acid-binding protein (I-FABP), soluble CD14 (sCD14), sCD163, IL-6, and sTNFRII were analyzed as markers of enterocyte turnover, inflammation, and immune activation. Non-parametric tests were used for analysis. Results 52 participants born in 14 countries were enrolled February–March 2019. Median CD4 count was 464/uL [95% CI 315–598]. Fourteen (27%) were positive for Strongyloides IgG. Strongyloides IgG levels correlated positively with sCD14 levels [r=0.36; P = 0.008]. Strongyloides + participants had significantly higher sCD14 levels compared with Strongyloides− participants [1.67 vs. 1.48 μg/mL, P = 0.031]. Among the Strongyloides + participants, Strongyloides IgG levels correlated with sCD163 levels [r=0.65, P = 0.026]. There were no difference in the other biomarkers. Logistical regression analysis showed that predictors of Strongyloides + include absolute eosinophil count (AEC) (OR 1.45 for every 100 increase of AEC [95% CI: 1.02, 2.15; P = 0.047]). CD4 count, number of years living in the United States, country of origin, and years from HIV diagnosis were not associated with test positivity. Conclusion Strongyloides co-infection is common among foreign-born PWH and may contribute to chronic monocyte/macrophage activation, a predictor of morbidity and mortality in PWH. Future directions include stool PCR confirmation of these infections, continued enrollment, and follow-up assays 6 months after treatment of Strongyloides to determine the impact on inflammation and risk of co-morbidities. Disclosures All authors: No reported disclosures. [ABSTRACT FROM AUTHOR]

Background Molecular profiling of tumour tissue to guide treatment of patients (pts) with advanced solid tumours has previously been reported. Most series were reported from large volume academic cancer centres in the West. There is a paucity of data reporting the experience and applicability of such an approach in Asia. We report our single-centre experience of conducting a prospective molecular profiling programme for pts in advanced solid tumours. Methods Pts with advanced solid tumours aged >/= 18 years, good performance status and archival tumour tissue available were prospectively consented. The RNA extracted from the FFPE tissue sections were subjected to RNA-sequencing using Archer FusionPlex Solid Tumor Kit and Archer Analysis Software 5.1. The putative gene translocations or alternative splicing were further confirmed with RT-PCR or FISH. Results 253 pts consented between Feb 2017 and Feb 2019. Demographics: M:F 138 : 115; Median age = 58 (range: 19-89). Diseases included sarcomas (n = 88, 35%), NSCLC (n = 61, 24%), glioma (n = 42, 17%), melanoma (n = 7, 3%) and others (n = 55, 21%). Median turn-around time from consent to availability of report: 24 days (range: 3-112 days). 89% of samples passed quality control assessments. 8% (n = 21) of samples could not be processed due to insufficient material (n = 18), decalcified specimen (n = 2) and poor RNA quality (n = 1). 20 pts (9% of tested samples) had actionable genomic alterations identified that resulted in off-label use of directed anti-cancer therapies, referral to clinical trials or compassionate access programmes. These include EGFRvIII mutations (n = 7) and MET alterations (n = 2) in gliomas; ROS1 (n = 4), ALK (n = 2), MET exon 14-skipping (n = 1) and RET (n = 1) rearrangements in NSCLC; ALK rearranged sarcoma (n = 1). Across all diseases, 3 pts with NTRK fusions were identified. Conclusions We report the first institutional-based molecular profiling programme in Hong Kong. Such programmes are feasible in Asia. Pts can potentially benefit from identification of actionable alterations within a reasonable time frame and be channelled to appropriate therapies or clinical trials. Legal entity responsible for the study The Chinese University of Hong Kong. Funding F. Hoffmann-La Roche Ltd (formerly Ignyta Inc.). Disclosure H. Loong: Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): Roche; Advisory / Consultancy: Celgene; Advisory / Consultancy, Research grant / Funding (institution), Travel / Accommodation / Expenses: Eli Lilly; Advisory / Consultancy, Travel / Accommodation / Expenses: Boehringer Ingelheim; Advisory / Consultancy, Research grant / Funding (institution), Travel / Accommodation / Expenses: Merck Sereno; Advisory / Consultancy, Speaker Bureau / Expert testimony: AbbVie; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution): Bristol-Myers Squibb; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Eisai; Speaker Bureau / Expert testimony: Guardant Health; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Novartis; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Bayer; Research grant / Funding (institution), Travel / Accommodation / Expenses: Merck Sharp & Dohme; Travel / Accommodation / Expenses: Pfizer; Advisory / Consultancy: Takeda ; Research grant / Funding (institution): Mundipharma. B.B.Y. Ma: Speaker Bureau / Expert testimony: Roche. E.P. Hui: Speaker Bureau / Expert testimony: Merck Sharp & Dohme; Speaker Bureau / Expert testimony: Merck Sereno; Advisory / Consultancy: Merck Sharp & Dohme; Research grant / Funding (institution): Merck Sharp & Dohme; Research grant / Funding (institution): Pfizer. All other authors have declared no conflicts of interest. [ABSTRACT FROM AUTHOR]

Background Around 30% of non-small cell lung cancer (NSCLC) patients harbor sensitizing mutations in the epidermal growth factor receptor (EGFR) gene. Most common EGFR mutations are exon 19 deletions and EGFR-L858R, that comprise 85% of EGFR-mutated cases, and whose drug sensitivity profile is already known. However, rare EGFR mutations represent 10-15% of EGFR-mutated NSCLCs with diverse sensitivity to different EGFR inhibitors. Methods Scientific literature and database search were conducted to identify pathogenicity and drug sensitivity of rare and common EGFR mutations. Structured data were uploaded in the database of the RealTime Oncology Treatment CalculatorTM (Calculator) to enable automated interpretation of variants. A structured database of clinical trials testing EGFR targeted therapies was also established for matching trial search. Molecular diagnostics of an NSCLC patient were performed (next-generation sequencing of 58 genes and FISH analyses) for therapeutic decision support. A rare EGFR mutation was identified (H773_V774insNPH) and automated classification was carried out using the Calculator. Results The Calculator is now able to automatically classify the most frequent 100+ EGFR mutations based on structured scientific evidence and by using a proprietary algorithm it prioritizes the associated EGFR inhibitors. The system can also handle complex molecular profiles so that resistance-associated alterations (e.g. KRAS mutation or ALK translocation) can be identified and optimal targeted therapies can be suggested. Matching clinical trials can be listed within seconds based on histology, biomarker criteria, compounds, previous therapies, etc. The NSCLC patient with the rare EGFR mutation (who had quickly progressed on gefitinib) had stable disease for 2 years on afatinib, which compound was suggested by the Calculator. Gefitinib was negatively associated with the molecular profile. Conclusions This case clearly demonstrates that knowledge on the drug sensitivity profile of the exact mutation is very important for the therapeutic decision. We demonstrate that RealTime Oncology Treatment CalculatorTM is an efficient tool for evidence-based prioritization of EGFR inhibitors even in the case of rare EGFR variants. Legal entity responsible for the study Oncompass Medicine Hungary Ltd. Funding National Research, Development and Innovation Office, National Oncogenomics and Precision Oncotherapy Program (NVKP_16-1-2016-0005), WESPED (KFI_16-1-2016-0048). Disclosure All authors have declared no conflicts of interest. [ABSTRACT FROM AUTHOR]

Background Structural variants in DNA such as translocations are increasingly found as key oncogenic events in tumors, and detection is clinically relevant for therapy choice in many conditions. Despite developments in targeted and whole genome gene sequencing, the robust detection of structural variants remains a challenge, especially in FFPE specimens. Methods Targeted proximity ligation assays selectively amplify and sequence entire genes based on the crosslinking of physically proximal sequences, and thereby enable complete sequencing of genes of interest, including single nucleotide and structural variants. Because these assays are based on the crosslinking and fragmenting of DNA, they have particular advantages in the analysis of FFPE samples, in which DNA is inherently crosslinked and fragmented. Results We will show that we have successfully developed targeted proximity ligation assays on representative clinical FFPE samples with considerably fragmented DNA. By making use of the DNA crosslinks in these samples, we can generate very broad sequence information over 100's of kb's, where other targeted NGS technologies are limited by the fragment size. We will show that we can detect NTRK fusions at DNA level, including determination of the fusion partner and the exact translocation breakpoint sequence. The latter can be used as a very sensitive and cancer-specific marker for minimal residual disease (MRD) testing and ctDNA monitoring with PCR. Conclusions Our newly developed targeted proximity ligation assays enable robust detection of structural variation in FFPE samples and the development of personalized MRD/ctDNA tests. Legal entity responsible for the study Cergentis. Funding Cergentis via Horizon2020 SME funding. Disclosure H. Feitsma: Full / Part-time employment: Cergentis. M. Yilmaz: Full / Part-time employment: Cergentis. J. Swennenhuis: Full / Part-time employment: Cergentis. A. Rakszewska: Full / Part-time employment: Cergentis. K. Hajo: Full / Part-time employment: Cergentis. E. Splinter: Full / Part-time employment: Cergentis. M. Simonis: Full / Part-time employment: Cergentis. M. Van Min: Shareholder / Stockholder / Stock options, Full / Part-time employment: Cergentis. All other authors have declared no conflicts of interest. [ABSTRACT FROM AUTHOR]

Background The ATP-binding cassette (ABC) transporters are superfamily proteins of cellular translocation machinery. Many ABC transporters release drugs to give multidrug resistance. However, ABC transporters existing in intracellular compartments give drug resistance by regulating biological processes, rather than direct release of drugs. The current study aimed to investigate the molecular mechanism that causes the drug resistance of the mitochondrial ABC transporters in cancer cells. Methods RNA-sequencing and bioinformatics analyses were performed to identify key mitochondrial ABC transporters and ABC transporter subfamily B member 7 (ABCB7)-mediated transcriptomic alterations. We investigated role of ABCB7 in iron homeostasis by examining iron concentration and reactive oxygen species (ROS) generation. Drug resistance and cellular survival mechanisms were understood by comparing the type and degree of cell death according to the ABCB7 expression. Results ABCB7 is highly expressed in patients with glioblastoma and has a negative correlation with prognosis. The increase in ABCB7 gives resistance to drug and the loss of ABCB7 leads to cell death. ABCB7 increases the expression of hypoxia-inducible factor 1 alpha (HIF1α) and activates HIF1α signaling by regulating iron concentration. When ABCB7 expression is maintained high in cancer cells, hypoxia-independent activation of HIF1α inhibits cell death by suppressing leucine zipper downregulated in cancer 1 (LDOC1). However, when ABCB7 is decreased, non-apoptotic cell death is induced by the change of iron concentration, and apoptosis which is inhibited by HIF1α signal is also induced. Additionally, because ABCB7 is essential for survival of universal cancer cells, it is presented as a pan-cancer core fitness gene. Conclusions Our findings demonstrate the ABCB7 inhibits both apoptotic and non-apoptotic cell death via maintenance of iron homeostasis and hypoxia-independent HIF1α signaling. Editorial acknowledgement Cancer Growth Regulation Laboratory for their discussion and technical assistance. Legal entity responsible for the study The authors. Funding National Research Foundation of Korea (NRF) [grant numbers: 2015R1A5A1009024 and 2017R1E1A1A01074205], the School of Life Sciences and Biotechnology for BK21 Plus, Korea University and Institute of Animal Molecular Biotechnology Korea University. Disclosure All authors have declared no conflicts of interest. [ABSTRACT FROM AUTHOR]

Background One of the most frequently diagnosed oncological diseases among young persons is Low-grade glioma (LGG). More than 80% of LGGs progress to higher-grade tumors always lethal to a patient. LGG is an extremely heterogeneous group of primary glial brain tumors with a quite variable rate of recurrence, survival, and response to treatment. Thus a study of new biomarkers associated with disease progression and patient outcome is useful to improve patient management. Genes involved in key cellular processes, e.g. apoptosis, are promising prognostic and predictive cancer markers. Phosphatidylserine (PS) plays an essential role in apoptosis signaling. Several studies suggested PS is dysregulated in the tumor microenvironment and offends the development of anti-cancer immune response. The scramblases PLSCR1 and XKR8 allegedly could be associated with the progression and outcome of LGG since they could be involved in the development of immunity via regulation of PS activity by its bidirectional translocation in the plasma membrane. Thus this study was aimed to evaluate associations of PLSCR1 and XKR8 with disease-free and overall survival of LGG patients. Methods Gene expression values and clinical information of 530 LGG patients were downloaded from TCGA Low-Grade Glioma study using the public resource CBioPortal. According to gene expression values (RNA Seq V2 RSEM) patients were divided into two groups: with downregulated (< mean) or upregulated (> mean) PLSCR1 or XKR8 gene. The Kaplan-Meier procedure and Log-Rank test were used for survival analysis. All calculations were performed in the integrated development environment - RStudio. Results It was shown the LGG patients with upregulated PLSCR1 (p = 2.54e-14) and XKR8 (p = 7.33e-15) gene had worse disease prognosis than other patients. Similar associations were seen in the study of LGG progression, where patients with upregulated PLSCR1 (p = 7.75e-07) and XKR8 (p = 5.59e-10) gene had a higher risk of cancer recurrence. Conclusions Consequently, PLSCR and XKR8 genes could be considered as new prognostic biomarkers of LGG progression and outcome. Legal entity responsible for the study Research Laboratory "Biomarker". Funding The work was performed according to the Russian Government Program of Competitive Growth of KFU. Disclosure All authors have declared no conflicts of interest. [ABSTRACT FROM AUTHOR]

Background In the United States, the most commonly transplanted tissues are bones, tendons, skin, heart valves, vessels and corneas. Over two million tissue grafts are distributed each year.Efforts to advance tissue transplant safety are less defined compared with organ transplant, and continuously being improved and regulated by national agencies. Environmental contamination, postmortem translocation of microorganisms, cross-contamination, and tissue processing are critical components which can be prevented with hand hygiene education thus preventing the transmission of infectious diseases. Microorganisms from technicians' skin can potentially result in infection and nontransplantable tissue. This is particularly true of regenerative tissue, such as Vivigen, that undergo no sterilization process. Methods A review was done of all 2018 ViviGen culture with these organisms: Bacillus, Cornebacterium, Diptheroids, Propionibacterium, Staphylococcus Coagulase negative, and Streptococcus viridians defined as Category 1. Results In 2018, 384 out of 1418 ViviGen donors were discarded due to positive cultures. Of the 384, 190 were due the presence of a Cat. 1 microorganism in post-processing cultures, for a discard rate of 17.6%. In these donors, preprocessing cultures were initially negative, thus indicating contamination was due to technician contamination. In the first quarter of 2019, and since implementation of Hand Hygiene education, we have already start to see a decline in acquisition of Cat 1 organism for a total reduction of 6% from Q1 of 2018 compared with 2019. Conclusion Tissue banking is a complex process of many steps to assure quality and safety of the tissue. Contamination is critical even when the microorganisms are considered low-virulent skin flora (i.e. Cat 1). This is especially important if these Cat 1 organisms are found in Post-processing cultures of grafts not terminally irradiated or sterilized. Implementation of proper hand hygiene education and aseptic techniques among tissue recovery and processing staff can significantly reduce the bioburden. Furthermore, a reduction in bioburden equates to higher likelihood in honoring the gift of donation in this selfless act of the donor and their loved one while also decreasing the financial loss due to culture results. Disclosures All authors: No reported disclosures. [ABSTRACT FROM AUTHOR]