miRNA 146b mediates the regulation of nucleolar size and activity in polyploid megakaryocytes.

BACKGROUND INFORMATION: Megakaryocytes (MKs) follow a unique cell cycle duplication process, called endomitosis, resulting in polyploidisation of cells. It is hypothesised that polyploidy, as well as an increment in cytoplasm volume, allow more efficient platelets generation from MKs. Although polyploidy leads to an increase in the DNA amount, which impacts gene expression, little is known about ribosomal biogenesis in these polylobulated polyploid cells. RESULTS: The nucleolus acts as a hub for ribosomal biogenesis, which in turn governs the protein synthesis rate of the cells. We therefore estimated the size and activity of the nucleolus in polyploid cells during megakaryopoiesis in vitro. Polyploid megakaryocytic cell lines and in vitro cultured MKs, which were obtained from human cord blood-derived CD 34+ cells, revealed that miRNA 146b regulated the activity of nucleolar and coiled-body phosphoprotein 1, which plays an integral role in determining nucleolar size and activity. Additionally, miRNA-146b was up-regulated during endomitosis and was found to promote megakaryopoiesis. CONCLUSION: We propose that miRNA 146b regulates not only nucleolar size and activity, but also megakaryopoiesis. SIGNIFICANCE: This study highlights the importance of nucleolar activity and miRNA in the progression of megakaryopoiesis and thrombopoiesis.

Palliative care provision at a tertiary cancer center during a global pandemic.

COVID-19 was first reported in Wuhan, China, in December 2019; it rapidly spread around the world and was declared a global pandemic by the World Health Organization in March 2020. The palliative care program at the Princess Margaret Cancer Centre, Toronto, Canada, provides comprehensive care to patients with advanced cancer and their families, through services including an acute palliative care unit, an inpatient consultation service, and an ambulatory palliative care clinic. In the face of a global pandemic, palliative care teams are uniquely placed to support patients with cancer who also have COVID-19. This may include managing severe symptoms such as dyspnea and agitation, as well as guiding advance care planning and goals of care conversations. In tandem, there is a need for palliative care teams to continue to provide care to patients with advanced cancer who are COVID-negative but who are at higher risk of infection and adverse outcomes related to COVID-19. This paper highlights the unique challenges faced by a palliative care team in terms of scaling up services in response to a global pandemic while simultaneously providing ongoing support to their patients with advanced cancer at a tertiary cancer center.

Clinical and epidemiological profile of Indian COVID-19 patients from Jaipur: a descriptive study.

We analyzed the data of 102 confirmed patients with novel Coronavirus 2 infection (COVID-19) during the early period of nationwide lockdown announced in India after the declaration of pandemic. We analyzed epidemiological, clinical characteristics and outcome of hospitalization in 102 patients with positive results for novel corona virus (SARS-CoV-2) RNA testing which were traced on the basis of history of travel, contact with a confirmed COVID-19 case, resident of hotspot areas or presence of symptoms, thus providing an accurate estimate of the proportion of asymptomatic cases in the initial population. Of 102 patients enrolled in the study, 83.3% (85/102) were asymptomatic and 16.67% (17/102) were symptomatic. Seventy-seven (75.49%) were males and 24.50% (25/102) were females. Eighteen (17.6%) patients had associated comorbidities, the most prevalent of which were diabetes mellitus 10.8% (11/102), hypertension 7.8% (8/102), chronic obstructive pulmonary disease (COPD) in 3.92% (4/102), chronic kidney Disease (CKD) 0.98% (1/102), coronary artery Disease (CAD) 0.98% (1/102) and cerebro-vascular disease (CVD) 0.98% (1/102). The clinical spectrum among symptomatic COVID-19 patients varied from dry cough and fever to respiratory failure and multi-organ failure. Twelve (11.76%) patients were kept in intensive care unit (ICU). Ninety-nine (97.05%) patients recovered while three (2.94%) died during hospital stay. With majority of COVID-19 cases in India being asymptomatic, changes in biochemical and inflammatory profile were small and insignificant in asymptomatic patients when compared to symptomatic patients. Elevated NLR, lymphopenia, age and presence of comorbidities were associated with increased severity and poor outcome.

Microvesicles promote megakaryopoiesis by regulating DNA methyltransferase and methylation of Notch1 promoter.

Megakaryopoiesis is the process of formation of mature megakaryocytes that takes place in the bone marrow niche resulting in the release of platelets into the peripheral blood. It has been suggested that cell to cell communication in this dense bone marrow niche may influence the fate of the cells. Numerous studies point to the role of exosomes and microvesicles not only as a messenger of the cellular crosstalk but also in growth and developmental process of various cell types. In the current study, we explored the effects of megakaryocyte-derived microvesicles in hematopoietic cell lines in the context of differentiation. Our study demonstrated that microvesicles isolated from the induced megakaryocytic cell lines have the ability to stimulate noninduced cells specifically into that particular lineage. We showed that this lineage commencement comes from the change in the methylation status of Notch1 promoter, which is regulated by DNA methyltransferases.

Clinical Profile of Covid-19 Infected Patients Admitted in a Tertiary Care Hospital in North India.

BACKGROUND: The novel coronavirus (Covid-19) continues to wreck havoc across China, European countries, USA and now seems to gain a strong foothold in India. The aim of this report is to describe the clinical profiles of these Covid-19 infected patients admitted in Sawai Mansingh Hospital(S.M.S), Jaipur ranging from their age, sex, travel history, clinical symptoms, laboratory evaluation, radiological characteristics, treatment provided along with common side effects and the final outcome. The described cases are one of the earliest cases of Covid-19 in the Indian subcontinent. METHODS: Epidemiological, clinical, laboratory, and radiological characteristics and treatment and outcomes data were obtained with data collection forms from electronic medical records and history given by 21 Covid-19 infected patients admitted in S.M.S., Jaipur. Patients were tested for Covid-19 by real-time reverse transcription polymerase chain reaction (RT-PCR) assay of 2019-nCoVRNA. RESULTS AND DISCUSSION: During the course of this study 21 Covid-19 positive patients were admitted in S.M.S Hospital, Jaipur. Male patients constituted 66.66% of total patients and majority of the patients (80.90%) were below 60 years of age. Most of the patients (71.40%) were either foreigners or had a history of foreign travel suggesting that these cases were not community acquired except for 4 cases from textile producing district Bhilwara (known as Manchester of India), an epicenter of North India. Approximately 33.33% patients were completely asymptomatic and of those who were symptomatic cough was the most common symptom (85.71%) followed by fever (78.57%), myalgia (64.28%), headache (28.57%) and dyspnea (28.57%). Three patients (14.28 %) had underlying co morbidity in the form of hypertension, diabetes mellitus, hypothyroidism, chronic kidney disease or coronary artery disease. 11 patients (52.38%) had lymphopenia in their hemogram during the course of admission. 3 patients (14.28%) had leucocytosis and 4 patients (19.04%) presented with thrombocytopenia. All 4 patients in the severe category had raised FDP, D-Dimer levels and they needed oxygen support. These patients had deranged liver functions and had elevated pro-calcitonin levels, serum ferritin levels and LDH levels. 1 out of the these 4 cases went into ARDS during the course of treatment. 10 patients yielded negative results for Covid-19. The mean duration from admission to getting 1st Covid-19 sample negative was 8.3 days. 18 patients (85.71%) are still under treatment. CONCLUSION: Clinical investigations in initial Covid-19 patients in the Indian subcontinent reveal lymphopenia as predominant finding in hemogram. Patients with older age and associated comorbid conditions (COPD and diabetes) seem to have greater risk for lung injury thereby requiring oxygen support during the course of disease and these patients also had greater derangement in their biochemical profile.

Alteration of mitochondrial proteome due to activation of Notch1 signaling pathway.

The Notch signaling pathway, a known regulator of cell fate decisions, proliferation, and apoptosis, has recently been implicated in the regulation of glycolysis, which affects tumor progression. However, the impact of Notch on other metabolic pathways remains to be elucidated. To gain more insights into the Notch signaling and its role in regulation of metabolism, we studied the mitochondrial proteome in Notch1-activated K562 cells using a comparative proteomics approach. The proteomic study led to the identification of 10 unique proteins that were altered due to Notch1 activation. Eight of these proteins belonged to mitochondria-localized metabolic pathways like oxidative phosphorylation, glutamine metabolism, Krebs cycle, and fatty acid oxidation. Validation of some of these findings showed that constitutive activation of Notch1 deregulated glutamine metabolism and Complex 1 of the respiratory chain. Furthermore, the deregulation of glutamine metabolism involved the canonical Notch signaling and its downstream effectors. The study also reports the effect of Notch signaling on mitochondrial function and status of high energy intermediates ATP, NADH, and NADPH. Thus our study shows the effect of Notch signaling on mitochondrial proteome, which in turn affects the functioning of key metabolic pathways, thereby connecting an important signaling pathway to the regulation of cellular metabolism.

p27 and leukemia: cell cycle and beyond.

Cell division is the foundation to development and the regulation of cell cycle progression is therefore of paramount importance to the living organisms. Primary control of cell cycle is achieved by an array of cyclins and cyclin dependent kinases (CDKs). The functions of these cyclin-CDK complexes are again regulated by a host of cyclin dependent kinase inhibitors (CDKI). Till date CDKIs are broadly classified into two groups-INK4 family (p15, p16, p18, and p19) and the cip/kip family (p21, p27, and p57). Collectively these CDKIs regulate the progression from G1 to S phase of cell cycle. This review summarizes the functions of p27 while highlighting its emerging roles in leukemia.

Molecular cross talk between Notch1, Shh and Akt pathways during erythroid differentiation of K562 and HEL cell lines.

Erythropoiesis is a tightly regulated process dependent on extrinsic signals conveyed by the bone marrow niche. The signalling pathways thus activated or repressed do not act in isolation; rather an intricate cross talk among these pathways ensues homoeostasis within the erythroid compartment. In this study, we describe the effects of two such signalling pathways namely the Notch1 and the Shh pathway on erythropoiesis in immortalised K562 and HEL cell lines as well as the cross talk that ensues between them. We show that while activation of the Notch1 pathway inhibits differentiation of erythroid lineage cell lines as well as in in-vitro primary erythroid cultures from the human CD34(+) cells; Shh pathway favours erythroid differentiation. Further, the Notch1 pathway activates the Akt pathway and constitutively active Akt partially mimics the effect of Notch1 activation on erythropoiesis. Moreover, the Notch1, Akt and Shh pathways were found to cross talk with each other. In this process, activation of Notch1 was found to down regulate the Shh pathway independent of Akt activation. Significantly, Notch1 not only down regulated the Shh pathway, but also inhibited recombinant Shh mediated erythropoiesis. Our study thus reveals an intricate crosstalk among the Notch1, Shh and Akt pathways wherein Notch1 emerges as a key regulator of erythropoiesis.

The oncology palliative care clinic at the Princess Margaret Cancer Centre: an early intervention model for patients with advanced cancer.

Several recently published randomized controlled trials have demonstrated the benefits of early palliative care involvement for patients with advanced cancer. In the oncology outpatient setting, palliative care clinics are an ideal site for the provision of early, collaborative support, which can be maintained throughout the cancer trajectory. Despite this, access to ambulatory palliative care clinics is limited, even at tertiary cancer centres. Existing programs for outpatient palliative care are variable in scope and are not well described in the literature. We describe the development and expansion of an outpatient palliative care clinic at the Princess Margaret Cancer Centre, Toronto, Canada, demonstrating how the clinic functions at a local and regional level. This clinic served as the intervention for a recent large cluster-randomized trial of early palliative care. The model for this service can be adapted by other palliative care programs that aim to provide early, integrated oncology care.

Epstein-Barr virus latent membrane protein-2A alters mitochondrial dynamics promoting cellular migration mediated by Notch signaling pathway.

Recently, migration and invasion of breast cancer cells have been linked with dysregulated mitochondrial dynamics. Mitochondria are essential cellular organelles that undergo continuous dynamic cycles of fission and fusion. It has been proposed that a delicate balance between these two processes is important for many pathophysiological outcomes including cancer. Epstein-Barr virus (EBV) is a gamma herpesvirus that is associated with various lymphoid and epithelial malignancies. The viral latent membrane protein 2A (LMP2A) has been shown to increase the invasive ability and induce epithelial-mesenchymal transition in nasopharyngeal carcinoma. Our present study reveals that mitochondrial dynamics also plays a critical role in Epstein-Barr virus-associated epithelial cancers. Our data indicate that viral LMP2A causes an elevated mitochondrial fission in gastric and breast cancer cells, which is manifested by elevated fission protein dynamin-related protein 1 (Drp1). Furthermore, LMP2A-mediated Notch pathway is responsible for this enhanced fission since inhibitors of the pathway decrease the expression of Drp1.

Differential regulation of MCM7 and its intronic miRNA cluster miR-106b-25 during megakaryopoiesis induced polyploidy.

Megakaryocytes exit from mitotic cell cycle and enter a phase of repeated DNA replication without undergoing cell division, in a process termed as endomitosis of which little is known. We studied the expression of a DNA replication licensing factor mini chromosome maintenance protein 7 (MCM7) and its intronic miR-106b-25 cluster during mitotic and endo-mitotic cycles in megakaryocytic cell lines and in vitro cultured megakaryocytes obtained from human cord blood derived CD34(+) cells. Our results show that contrary to mitotic cell cycle, endomitosis proceeds with an un-coupling of the expression of MCM7 and miR-106b-25. This was attributed to the presence of a transcript variant of MCM7 which undergoes nonsense mediated decay (NMD). Additionally, miR-25 which was up regulated during endomitosis was found to promote megakaryopoiesis by inhibiting the expression of PTEN. Our study thus highlights the importance of a transcript variant of MCM7 destined for NMD in the modulation of megakaryopoiesis.

Adaptive fractional-order sliding-mode disturbance observer-based robust theoretical frequency controller applied to hybrid wind-diesel power system.

This work presents design and theoretical analysis of an adaptive fractional-order sliding-mode disturbance observer (FO-SM-DOB)-aided fractional-order robust controller for frequency regulation of a hybrid wind-diesel based power system, considering endogenous/exogenous system disturbances. Adaptive FO-SM-DOB is designed to estimate unknown/uncertain lumped system disturbances, including parametric uncertainty and exogenous disturbances. Afterwards, an improved fractional-order sliding mode controller (FOSMC) augmented with the estimated output of FO-SM-DOB is designed and applied to accelerate system dynamics with minimum chattering in the control effort. The Mittag-Leffler stability theorem affirms the finite-time convergence of disturbance estimation error. Moreover, the closed-loop asymptotic stability of the overall control system has been guaranteed by applying Lyapunov argument. The effectiveness of the suggested resilient fractional-order nonlinear frequency controller is theoretically validated by performing an extensive comparative study with SMC, FOSMC (without DOB), state observer-based SMC (SOB-SMC), second-order SMC (without DOB), and conventional integer/fractional-order controllers. Simulation results establish the supremacy of the proposed resilient fractional-order nonlinear frequency controller over its other counterparts concerning fast disturbance rejection, weaker chattering, and a high degree of robustness against unknown lumped system disturbances. Further, to demonstrate the practicability and validate the effectiveness of the proposed control strategy, magnetic levitation system and IEEE 39-bus New England power system are considered and successfully tested on MATLAB platform.

Family physicians' involvement in palliative cancer care.

BACKGROUND: Family physicians' (FPs) long-term relationships with their oncology patients position them ideally to provide primary palliative care, yet their involvement is variable. We examined perceptions of FP involvement among outpatients receiving palliative care at a cancer center and identified factors associated with this involvement. METHODS: Patients with advanced cancer attending an oncology palliative care clinic (OPCC) completed a 25-item survey. Eligible patients had seen an FP within 5 years. Binary multivariable logistic regression analyses were conducted to identify factors associated with (1) having seen an FP for palliative care within 6 months, and (2) having a scheduled/planned FP appointment. RESULTS: Of 258 patients, 35.2% (89/253) had seen an FP for palliative care within the preceding 6 months, and 51.2% (130/254) had a scheduled/planned FP appointment. Shorter travel time to FP (odds ratio [OR] = 0.67, 95% confidence interval [CI] = 0.48-0.93, p = 0.02), the FP having a 24-h support service (OR = 1.96, 95% CI = 1.02-3.76, p = 0.04), and a positive perception of FP's care (OR = 1.05, 95% CI = 1.01-1.09, p = 0.01) were associated with having seen the FP for palliative care. English as a first language (OR = 2.90, 95% CI = 1.04-8.11, p = 0.04) and greater ease contacting FP after hours (OR = 1.33, 95% CI = 1.08-1.64, p = 0.008) were positively associated, and female sex of patient (OR = 0.51, 95% CI = 0.30-0.87, p = 0.01) and travel time to FP (OR = 0.66, 95% CI = 0.47-0.93, p = 0.02) negatively associated with having a scheduled/planned FP appointment. Number of OPCC visits was not associated with either outcome. CONCLUSION: Most patients had not seen an FP for palliative care. Accessibility, availability, and equity are important factors to consider when planning interventions to encourage and facilitate access to FPs for palliative care.

Investigating the potential molecular players and therapeutic drug molecules in carfilzomib resistant multiple myeloma by comprehensive bioinformatics analysis.

Multiple myeloma (MM), second most common hematological malignancy, still remains beyond cure because of acquirement of drug resistance. Proteasome inhibitor such as carfilzomib (Cfz) therapy which has been used as one of the key therapies against MM recently, is obstructed by the incidence of Cfz resistance. The underlying mechanism of this acquired Cfz resistance in MM is very little understood. Therefore, the current study was aimed to investigate the differentially expressed genes (DEGs), associated micro RNAs (miRNAs), and transcription factors (TFs) from the microarray datasets of Cfz resistant and Cfz sensitive MM cell lines, obtained from Gene Expression Omnibus (GEO) database. DEGs were detected using GEO2R tool from two datasets and common DEGs were identified through Venn diagram. Gene ontology (GO) and pathway enrichment analysis were performed on DAVID database. Through STRING database and Cytoscape tool, protein-protein interaction (PPI) network of DEGs was constructed. Genetic alterations in DEGs were investigated using COSMIC database. Interaction network between DEGs and miRNAs as well as TFs were obtained and constructed by using mirDIP, TRRUST, and miRNet tools. Drug gene interaction analysis was performed to identify potential drug molecules on DGIdb tool. Several common DEGs were identified in Cfz resistant MM. PDGF, VEGF, and Wnt signaling pathways were significantly enriched and might be involved in MM progression. miRNA analysis identified hsa-mir-124-3p, hsa-mir-26a-5p that can target DEGs. Various drug molecules such as dabrafenib, vemurafenib, and venetoclax that could potentially attenuate the MM pathophysiology, were detected. The entire study might provide a new understanding about the Cfz resistance in MM.

In silico bioinformatics analysis for identification of differentially expressed genes and therapeutic drug molecules in Glucocorticoid-resistant Multiple myeloma.

Multiple myeloma (MM), second most common hematological malignancy, still remains irremediable because of acquisition of drug resistance. Glucocorticoid (GC) therapy, which is used as one of the key therapies against MM, is hindered by the incidence of GC resistance. The underlying mechanism of this acquired GC resistance in MM is not fully elucidated. Therefore, the present study was aimed to identify the differentially expressed genes (DEGs), associated micro RNAs (miRNAs), and transcription factors (TFs) from the microarray datasets of GC-resistant and GC-sensitive MM cell lines, obtained from Gene Expression Omnibus (GEO) database. DEGs were identified using GEO2R tool from two datasets and common DEGs were obtained by constructing Venn diagram. Then the Gene ontology (GO) and pathway enrichment analysis were performed using DAVID database. Genetic alterations in DEGs were examined using COSMIC database. Protein-protein interaction (PPI) network of DEGs was constructed using STRING database and Cytoscape tool. Network of interaction of DEGs and miRNAs as well as TFs were obtained and constructed using mirDIP, TRRUST, and miRNet tools. Drug gene interaction was studied to identify potential drug molecules by DGIdb tool. Six common DEGs, CDKN1A, CDKN2A, NLRP11, BTK, CD52, and RELN, were found to be significantly upregulated in GC-resistant MM and selected for further analysis. miRNA analysis detected hsa-mir-34a-5p that could interact with maximum target DEGs. Two TFs, Sp1 and Sp3, were found to regulate the expression of selected DEGs. The entire study may provide a new understanding about the GC resistance in MM.

Performance evolution of different controllers for frequency regulation of a hybrid energy power system employing chaotic crow search algorithm.

The work described herein compares the performance of different optimized controllers, viz. proportional-integral, proportional-integral-derivative (PID) with filter, two-degree-of-freedom (2DOF)-PID, 3DOF-PID, fractional-order-PID, cascade PI-PID, tilt-integral-derivative (TID), and cascade-TID (CC-TID) controllers in frequency regulation of a hybrid energy distributed power system (HEDPS). The HEDPS is integrated with a multi-unit hydrothermal power plant for ensuring stable power supply. Crow search algorithm has been adopted with chaotic mapping (CCSA) for fine-tuning of the controller settings mentioned above. Extensive analysis has been presented to confirm the superiority of the CC-TID controller compared to other prevalent controllers of state-of-art in terms of different performance specifications. The tuning competence of the CCSA has been demonstrated over conventional CSA and other available optimization techniques. To enhance the mastery of the controller, disturbance-observer (Dob) is developed to estimate fast-changing disturbance profiles and subsequently refines the control law. The controller's robustness is affirmed under random perturbations, presence of nonlinearities, and variation of parameters. The effect of integration of a geothermal power plant on the system performance has also been outlined. The efficacy of Dob-aided CC-TID controller in frequency regulation is validated thereof.

DNA binding domain of RFX5: interactions with X-box DNA and RFXANK.

Regulatory factor X (RFX) is a heterotrimeric protein complex having RFX5, RFXANK and RFXAP as its three subunits. It is involved in the regulation of the transcription of MHCII molecules in antigen presenting cells. The RFX complex binds to X-box DNA, using the DNA binding domain, present in RFX5. The DNA binding domain (DBD) of RFX5 (12kD) and intact RFXANK (35 kD) were subcloned, expressed and purified. The associations of RFX5DBD with the X-box DNA and between RFX5DBD and RFXANK were measured in this study. The interaction of RFX5DBD and X-box DNA was studied using steady state fluorescence quenching and circular dichroism. The binding dissociation constant (K(d)) of the DNA-protein complex was determined from fluorescence measurements. The van't Hoff plot was linear over the temperature range 10-25 degrees C and the binding was found to be entropy-driven and enthalpy-favorable. The effect of electrolytes in RFX5DBD-DNA association was also studied. Molecular association between RFX5DBD and RFXANK has been observed by fluorescence resonance energy transfer (FRET) measurements, changes in the ratio of the two vibronic intensities of pyrene labeled RFX5DBD in presence of RFXANK and chemical cross-linking followed by tandem mass spectrometry. Results showed that the two proteins could interact in the absence of the third subunit RFXAP, in vitro with an apparent dissociation constant (K(d)) of 128 nM.

Outpatient palliative medicine consultations: urgent or routine?

BACKGROUND: Although outpatient palliative care clinics (OPCCs) provide a venue for early, pre-emptive referral to palliative care on a routine basis, some patients will continue to require urgent referrals. The purpose of this study was to characterise these urgent referrals to determine whether they reflect clinical need or convenience. METHODS: We retrospectively compared new patients in an OPCC who were seen urgently versus those seen at routine appointments. Descriptive statistics compared the two groups in terms of clinical characteristics, referring teams, symptoms, performance status and outcomes. Logistic regression was used to identify factors associated with urgent referral to the OPCC. Overall survival was compared using the log-rank test. RESULTS: Between January 2016 and December 2017, a total of 113 urgent referrals were reviewed in the OPCC; these were compared with a random sample of 217 routine referrals. Patients seen urgently were more likely to be referred by surgical oncology, and to report worse symptom scores for pain (p=0.0007), tiredness (p=0.02), well-being (p=0.001), constipation (p=0.02) and sleep (p=0.01). More patients seen urgently required direct admission to hospital following the visit (17.7% vs 0.9%, p<0.001). Median survival was shorter for patients seen urgently (4.3 months, 95% CI 3.4 to 7.8) versus routinely (8.1 months, 95% CI 7.2 to 9.5). CONCLUSIONS: Compared with routine referrals, new patients seen urgently in the OPCC had higher symptom burden, shorter median survival and a greater chance of direct admission to hospital. Palliative care clinics should consider how best to accommodate urgent referrals.

Palliative medicine outpatient clinic 'no-shows': retrospective review.

OBJECTIVES: Patients who do not attend outpatient palliative care clinic appointments ('no-shows') may have unmet needs and can impact wait times. We aimed to describe the characteristics and outcomes associated with no-shows. METHODS: We retrospectively reviewed new no-show referrals to the Princess Margaret Cancer Centre Oncology Palliative Care Clinic (OPCC) in Toronto, Canada, between January 2017 and December 2018, compared with a random selection of patients who attended their first appointment, in a 1:2 ratio. We collected patient information, symptoms, performance status (Eastern Cooperative Oncology Group (ECOG) and outcomes. Univariable and multivariable logistic regression analyses were used to identify significant factors. RESULTS: Compared with those who attended (n=214), no-shows (n=103), on multivariable analysis, were at higher odds than those who attended of being younger (OR 0.98, 95% CI 0.96 to 1.00, p=0.019), living outside Toronto (OR 2.67, 95% CI 1.54 to 4.62, p<0.001) and having ECOG ≥2 (OR 2.98, 95% CI 1.41 to 6.29, p=0.004). No-shows had a shorter median survival compared with those who attended their first appointment (2.3 vs 8.7 months, p<0.001). CONCLUSION: Compared with patients who attended, no-shows lived further from the OPCC, were younger, and had a poorer ECOG. Strategies such as virtual visits should be explored to reduce no-shows and enable attendance at OPCCs.

Downregulation of HLA-ABC expression through promoter hypermethylation and downmodulation of MIC-A/B surface expression in LMP2A-positive epithelial carcinoma cell lines.

Epstein Barr Virus (EBV) is a human herpesvirus, and has been reported to be associated with nasopharyngeal carcinoma, gastric carcinoma, Burkitt's lymphoma and Hodgkin's lymphoma. In most of the associated tumors, the virus remains in a latently infected state. During latency, EBV expresses Latent Membrane Protein 2A (LMP2A) along with few other genes. We previously showed that LMP2A causes downregulation of HLA-ABC surface expression in EBV associated gastric carcinomas. However, the mechanism that leads to this downregulation remain unclear. We therefore analyzed methylation-mediated regulation of HLA-ABC expression by LMP2A. Interestingly, according to the 'missing self' hypothesis, when there is a decrease in HLA-ABC surface expression, expression of NKG2D ligands' must be upregulated to facilitate killing by Natural Killer (NK) cells. Analysis of NKG2D ligands' expression, revealed downregulation of MIC-A/B surface expression in response to LMP2A. Furthermore, the role of Unfolded Protein Response (UPR) in the regulation of MIC-A/B surface expression in cells expressing LMP2A was also investigated. Protein Disulfide Isomerase (PDI) mediated inhibition of MIC-A/B surface expression was observed in LMP2A expressing cells. Our current findings provide new insights in LMP2A arbitrated dysregulation of host immune response in epithelial cell carcinomas.