14-3-3ζ binds the proteasome, limits proteolytic function and enhances sensitivity to proteasome inhibitors.

14-3-3 proteins are a family of master regulators of intracellular signaling, yet their impact on proteasome function is unknown. We demonstrate that 14-3-3ζ binds the 11S proteasome activator, limiting proteasome assembly and cellular capacity for protein degradation. To define the functional impact of 14-3-3ζ proteasomal binding in myeloma cells, silencing and overexpression experiments are performed. We find that downregulation of 14-3-3ζ impairs myeloma cell growth and confers resistance to clinically used proteasome inhibitors. In a large cohort of newly diagnosed myeloma patients, elevated expression of 14-3-3ζ is associated with high risk myeloma genetic subtypes and worse prognosis overall. Our work demonstrates the important role of 14-3-3ζ in regulating proteasome function, myeloma cell growth and sensitivity to therapeutics, and suggests regulation of 14-3-3ζ as a new approach in myeloma therapy.

Nucleotide excision repair is a potential therapeutic target in multiple myeloma.

Despite the development of novel drugs, alkylating agents remain an important component of therapy in multiple myeloma (MM). DNA repair processes contribute towards sensitivity to alkylating agents and therefore we here evaluate the role of nucleotide excision repair (NER), which is involved in the removal of bulky adducts and DNA crosslinks in MM. We first evaluated NER activity using a novel functional assay and observed a heterogeneous NER efficiency in MM cell lines and patient samples. Using next-generation sequencing data, we identified that expression of the canonical NER gene, excision repair cross-complementation group 3 (ERCC3), significantly impacted the outcome in newly diagnosed MM patients treated with alkylating agents. Next, using small RNA interference, stable knockdown and overexpression, and small-molecule inhibitors targeting xeroderma pigmentosum complementation group B (XPB), the DNA helicase encoded by ERCC3, we demonstrate that NER inhibition significantly increases sensitivity and overcomes resistance to alkylating agents in MM. Moreover, inhibiting XPB leads to the dual inhibition of NER and transcription and is particularly efficient in myeloma cells. Altogether, we show that NER impacts alkylating agents sensitivity in myeloma cells and identify ERCC3 as a potential therapeutic target in MM.

Protein arginine methyltransferase 5 has prognostic relevance and is a druggable target in multiple myeloma.

Arginine methyltransferases critically regulate cellular homeostasis by modulating the functional outcome of their substrates. The protein arginine methyltransferase 5 (PRMT5) is an enzyme involved in growth and survival pathways promoting tumorigenesis. However, little is known about the biologic function of PRMT5 and its therapeutic potential in multiple myeloma (MM). In the present study, we identified and validated PRMT5 as a new therapeutic target in MM. PRMT5 is overexpressed in patient MM cells and associated with decreased progression-free survival and overall survival. Either genetic knockdown or pharmacological inhibition of PRMT5 with the inhibitor EPZ015666 significantly inhibited growth of both cell lines and patient MM cells. Furthermore, PRMT5 inhibition abrogated NF-κB signaling. Interestingly, mass spectrometry identified a tripartite motif-containing protein 21 TRIM21 as a new PRMT5-partner, and we delineated a TRIM21-dependent mechanism of NF-κB inhibition. Importantly, oral administration of EPZ015666 significantly decreased MM growth in a humanized murine model of MM. These data both demonstrate the oncogenic role and prognostic relevance of PRMT5 in MM pathogenesis, and provide the rationale for novel therapies targeting PRMT5 to improve patient outcome.

Expressed fusion gene landscape and its impact in multiple myeloma.

Multiple myeloma is a plasma cell malignancy characterized by recurrent IgH translocations and well described genomic heterogeneity. Although transcriptome profiles in multiple myeloma has been described, landscape of expressed fusion genes and their clinical impact remains unknown. To provide a comprehensive and detailed fusion gene cartography and suggest new mechanisms of tumorigenesis in multiple myeloma, we performed RNA sequencing in a cohort of 255 newly diagnosed and homogeneously treated multiple myeloma patients with long follow-up. Here, we report that patients have on average 5.5 expressed fusion genes. Kappa and lambda light chains and IgH genes are main partners in a third of all fusion genes. We also identify recurrent fusion genes that significantly impact both progression-free and overall survival and may act as drivers of the disease. Lastly, we find a correlation between the number of fusions, the age of patients and the clinical outcome, strongly suggesting that genomic instability drives prognosis of the disease.

Blockade of deubiquitylating enzyme Rpn11 triggers apoptosis in multiple myeloma cells and overcomes bortezomib resistance.

Proteasome inhibition is an effective therapy for multiple myeloma (MM) patients; however, the emergence of drug resistance is common. Novel therapeutic strategies to overcome proteasome inhibitor resistance are needed. In this study, we examined whether targeting deubiquitylating (DUB) enzymes upstream of 20S proteasome overcomes proteasome inhibitor resistance. Gene expression analysis, immunohistochemical studies of MM patient bone marrow, reverse transcription-PCR and protein analysis show that Rpn11/POH1, a DUB enzyme upstream of 20S proteasome, is more highly expressed in patient MM cells than in normal plasma cells. Importantly, Rpn11 expression directly correlates with poor patient survival. Loss-of-function studies show that Rpn11-siRNA knockdown decreases MM cell viability. Pharmacological inhibition of Rpn11 with O-phenanthroline (OPA) blocks cellular proteasome function, induces apoptosis in MM cells and overcomes resistance to proteasome inhibitor bortezomib. Mechanistically, Rpn11 inhibition in MM cells activates caspase cascade and endoplasmic stress response signaling. Human MM xenograft model studies demonstrate that OPA treatment reduces progression of tumor growth and prolongs survival in mice. Finally, blockade of Rpn11 increases the cytotoxic activity of anti-MM agents lenalidomide, pomalidomide or dexamethasone. Overall, our preclinical data provide the rationale for targeting DUB enzyme Rpn11 upstream of 20S proteasome to enhance cytotoxicity and overcome proteasome inhibitor resistance in MM.

MUC1-C integrates PD-L1 induction with repression of immune effectors in non-small-cell lung cancer.

Immunotherapeutic approaches, particularly programmed death 1/programmed death ligand 1 (PD-1/PD-L1) blockade, have improved the treatment of non-small-cell lung cancer (NSCLC), supporting the premise that evasion of immune destruction is of importance for NSCLC progression. However, the signals responsible for upregulation of PD-L1 in NSCLC cells and whether they are integrated with the regulation of other immune-related genes are not known. Mucin 1 (MUC1) is aberrantly overexpressed in NSCLC, activates the nuclear factor-κB (NF-κB) p65→︀ZEB1 pathway and confers a poor prognosis. The present studies demonstrate that MUC1-C activates PD-L1 expression in NSCLC cells. We show that MUC1-C increases NF-κB p65 occupancy on the CD274/PD-L1 promoter and thereby drives CD274 transcription. Moreover, we demonstrate that MUC1-C-induced activation of NF-κB→︀ZEB1 signaling represses the TLR9 (toll-like receptor 9), IFNG, MCP-1 (monocyte chemoattractant protein-1) and GM-CSF genes, and that this signature is associated with decreases in overall survival. In concert with these results, targeting MUC1-C in NSCLC tumors suppresses PD-L1 and induces these effectors of innate and adaptive immunity. These findings support a previously unrecognized central role for MUC1-C in integrating PD-L1 activation with suppression of immune effectors and poor clinical outcome.

miR-23b/SP1/c-myc forms a feed-forward loop supporting multiple myeloma cell growth.

Deregulated microRNA (miR)/transcription factor (TF)-based networks represent a hallmark of cancer. We report here a novel c-Myc/miR-23b/Sp1 feed-forward loop with a critical role in multiple myeloma (MM) and Waldenstrom's macroglobulinemia (WM) cell growth and survival. We have found miR-23b to be downregulated in MM and WM cells especially in the presence of components of the tumor bone marrow milieu. Promoter methylation is one mechanism of miR-23b suppression in myeloma. In gain-of-function studies using miR-23b mimics-transfected or in miR-23b-stably expressing MM and WM cell lines, we observed a significant decrease in cell proliferation and survival, along with induction of caspase-3/7 activity over time, thus supporting a tumor suppressor role for miR-23b. At the molecular level, miR-23b targeted Sp1 3'UTR and significantly reduced Sp1-driven nuclear factor-κB activity. Finally, c-Myc, an important oncogenic transcription factor known to stimulate MM cell proliferation, transcriptionally repressed miR-23b. Thus MYC-dependent miR-23b repression in myeloma cells may promote activation of oncogenic Sp1-mediated signaling, representing the first feed-forward loop with critical growth and survival role in myeloma.

Evaluation of the usability of a serious game aiming to teach facial expressions to schizophrenic patients.

In some psychological disorders such as autism and schizophrenia, loss of facial expression recognition skill may complicate patient's daily life. Information technology may help to develop facial expression recognition skill by educational software and games. We designed and developed an interactive web-based educational program with which we performed a usability study before investigating its effectiveness on the schizophrenia patients' ability of emotion perception. The purpose of this study is to describe the usability evaluation for a web-based game set that has been designed to teach facial expressions to schizophrenic patients. The usability study was done at two steps; first, we applied heuristic evaluation and the violations were rated in a scale from most to least severe and the major problems were solved. In the second step, think-aloud method was used and the web site was assessed by five schizophrenic patients. Eight experts participated in the heuristic evaluation, in which a total of 60 violations were identified with a mean severity of 2.77 (range: 0-4). All of the major problems (severity over 2.5) were listed and the usability problems were solved by the development team. After solving the problems, five users with a diagnosis of schizophrenia used the web site with the same scenario. They reported to have experienced minor, but different problems. In conclusion, we suggest that a combination of heuristic evaluation and think-aloud method may be an effective and efficient way for usability evaluations for the serious games that have been designed for special patient groups.

Características clínicas de pacientes ingresados con diagnóstico de insuficiencia cardiaca en un hospital de la familia y la comunidad / Clinical characteristics of patients admitted with diagnosis of heart failure in a hospital of family and community

INTRODUCCIÓN: La Insuficiencia Cardiaca corresponde a una patología cardiovascular muy frecuente en nuestra población. Causa gran número de hospitalizaciones al año, significando un gasto importante en salud. Hay clara relación de factores demográficos con el desarrollo, evolución y pronóstico de IC, siendo uno de los más aceptados la edad, y uno de los más discutidos, el sexo. El objetivo de este trabajo ha sido caracterizar a los pacientes con insuficiencia cardiaca que se hospitalizaron en el Servicio de Medicina del Hospital de Familia y Comunidad de Mulchén, Chile, durante el año 2011. MATERIAL Y MÉTODO: Se realizó un estudio retrospectivo, observacional y descriptivo, mediante la revisión de los registros oficiales de ingreso-egreso del Servicio de Medicina del Hospital de Mulchén durante el 2011, analizando variables como edad, sexo y días de hospitalización. Se incluyó un total de 162 pacientes; 84 varones (51,85 por ciento). RESULTADOS: Las edades variaron entre 38 y 96 años, con un promedio de 67 años y con un 73,45 por ciento de pacientes con 60 años o más. La duración de la hospitalización varió entre 3 y 19 días, con un promedio de11. Casi el 80 por ciento de los pacientes provino del servicio de urgencia. Los diagnósticos asociados más frecuentes fueron hipertensión/cardiopatía hipertensiva (22 por ciento), y cardiopatía coronaria (11,2 por ciento). Se registraron 12 fallecimientos (7 por ciento). El 55 por ciento de los pacientes corresponde al Fondo Nacional de Salud B. DISCUSIÓN: Los resultados obtenidos están de acuerdo a lo descrito en la literatura para la población general y aportan datos útiles para enfrentar al paciente con insuficiencia cardiaca que se atiende en el servicio.

INTRODUCTION: Heart Failure is very frequent in our adult population. It causes a great number of hospitalizations a year and means an important expense in health. There is a clear relationship between diverse demographic factors and the development, evolution and prognosis of heart failure, being one of the accepted the age and of the discussed the sex. The objective of this study was to characterize hospitalized patients with heart failure in the Department of Medicine of the Familia y Comunidad Hospital of Mulchén, Chile, in 2011. MATERIAL AND METHOD: Retrospective, observational and descriptive study, there were checked the records of admission-discharge of the department and were selected all the patients with heart failure diagnosis at the income analyzing variables as age, sex, days and others. Included a total of 162 patients; 84 males (51.85 percent).RESULTS: The ages ranged from 38 to 96 years, with an average of 67 years, with a 73.45 percent of patients 60 years or older. The length of hospitalization ranged between 3 and 19 days, with an average of 11 days. Almost 80 percent of the patients came from to Emergency Department. The most frequent associated diagnoses were hypertension/hypertensive cardiomyopathy (22 percent) and coronary heart disease (11.2 percent. There were 12 deaths (7 percent). 55 percent of patients were affiliated to Fondo Nacional de Salud B.DISCUSSION: We conclude that the results obtained are concordant with those described in the literature for the general population and provide useful data to confront the patient with heart failure served in our service.

Características clínicas de adultos con diabetes mellitus 2 del consultorio del Hospital de Penco-Lirquén, Chile / Profile of adult patients with type-2 diabetes mellitus at the outpatient clinic of Penco-Lirquén Hospital, Chile

INTRODUCCIÓN: La Diabetes Mellitus es un conjunto de trastornos metabólicos cuya característica común es la hiperglicemia. Es importante destacar que dentro de sus complicaciones, la principal causa de muerte en el paciente diabético es cardiovascular. De este modo, un control adecuado de la patología y el manejo de los factores de riesgo asociados tienen una repercusión significativa en la morbimortalidad de este grupo de pacientes. OBJETIVO: Caracterizar clínicamente a los pacientes adultos con diabetes mellitus tipo 2 atendidos en el programa de salud cardiovascular del Consultorio del Hospital de Penco –Lirquén. MATERIAL Y MÉTODO: Estudio retrospectivo, observacional y descriptivo, revisando la última atención de los tarjetones de control crónico de los pacientes del programa de salud cardiovascular del Consultorio del Hospital de Penco-Lirquén, entre enero y diciembre del 2011, excluyéndose los adultos mayores(con edad ≥65 años). RESULTADOS: El 68,8 por ciento de los pacientes(n=86) eran mujeres. El promedio de edad fue de 52,88 años. El 58,1 por ciento presentó valores de presión arterial en rango de hipertensión.En relación a los valores de hemoglobina glicosilada, en el 63,8 por ciento fue mayor a 8. El promedio de índice de masa corporal fue de 32,4. DISCUSIÓN: La característica clínico-epidemiológica más destacada del estudio fue la mala compensación de los pacientes tanto en el control de diabetes mellitus como en los factores de riesgo asociados al desarrollo de patologías cardiovasculares, lo que requiere una intervención urgente para una mejor calidad de vida futura.

INTRODUCTION: Diabetes Mellitus is a group of metabolic disorders characterized by hyperglycemia that has increased in prevalence in the last years. Between its complications, we can find the main cause of death in diabetic patients: cardiovascular. Therefore, an adequate control and management of the associated risk factors can have a significant repercussion on the morbidity and mortality of this group of patients. OBJECTIVE: To characterize the adult patients with type 2 – diabetes mellitus, controlled at the cardiovascular health program of the Outpatient Clinic of the Penco-Lirquén Hospital, during 2011, excluding patients with 65 years old or more. MATERIAL AND METHOD: Retrospective, observational and descriptive study,by checking the last control registered in the chronic control cards of the patients of the cardiovascular health program of the Outpatient Clinic of the Penco-Lirquén Hospital, from January to December, 2011. RESULTS: 68.8 percent of the patients (n=86) were women. The average age was 52.88 years. 58.1 percent had blood pressure in hypertensive levels. The 63.8 percent had glycosylated hemoglobin above 8, and only 26 percent had normal cholesterol levels. DISCUSSION: The most important epidemiologic and clinic aspect of the study was the bad control of the patients, in the particular goals of diabetes mellitus, and in the risk factors associated with the development of cardiovascular diseases, situation that requires an urgent intervention for a better future quality of life.

A simple and accurate prediction model to estimate the intrahospital mortality risk of hospitalised cancer patients.

We aimed to form a risk prediction model to assess the probability of intrahospital death in cancer patients at the time of hospitalisation. The medical records and the relevant clinical parameters of cancer patients who died in or who were discharged from a teaching hospital between 1997 and 2000 (n = 334) were reviewed to explore the determinants of intrahospital death, which later were verified prospectively (n = 131). Eastern Cooperative Oncology Group (ECOG) performance status of four, short duration of disease (on a logarithmic scale), emergency admission, low haemoglobin (Hb) value (on a linear scale) and lactate dehydrogenase (LDH) value greater than 378 micro/ml were significantly and independently associated with the risk of intrahospital death. This model had a receiver operating characteristic area of 0.88 in the derivation cohort and 0.82 in the validation cohort. Using readily available clinical parameters, it is possible to devise an accurate and applicable risk prediction model for the hospitalised cancer patients.

The value of CEA and CA 19-9 in detecting cancer in a group of high-risk subjects with gastrointestinal symptoms.

The aim of this study was to evaluate the clinical value of CEA and CA 19-9 in a potential high-risk population of subjects with gastrointestinal complaints. The basic question was whether the determination of these markers, in addition to some other clinical features in this high risk population, could be helpful in diagnosing intraabdominal cancer. Two hundred and two patients with gastrointestinal complaints underwent standard diagnostic procedures and were followed for at least one year. For every patient, CEA and CA 19-9 levels were obtained at the first examination; the evaluating physician was blinded to the marker levels. The determinants of the likelihood of cancer were evaluated by multivariate analysis. Seventeen patients were diagnosed as having intraabdominal cancers. With the presence of melena (RR = 101.63, p = 0.007), nonspecific gastrointestinal symptoms (RR = 12.54, p = 0.026), increasing age (RR = 1.09, p = 0.028) and abnormal CEA (RR = 240.79, p = 0.000), the risk of having cancer increased significantly and independently. The presence of a primary gastric complaint was associated with a lower risk of cancer in this cohort (RR = 0.01, p = 0.04). Markers were not used in the diagnostic workup. In conclusion, in patients presenting with gastrointestinal complaints, the finding of elevated CEA levels may help in the diagnosis of cancer by prompting a more extensive search for intraabdominal cancer.

Does awareness of diagnosis make any difference to quality of life? Determinants of emotional functioning in a group of cancer patients in Turkey.

The object of this study was to investigate how the information status with regard to diagnosis, in addition to social and clinical factors, influenced emotional functioning and quality of life in a group of cancer patients in Turkey. A consecutive sample of 100 cancer patients being treated for different diagnoses in a tertiary care centre were prospectively evaluated. Data on patient disease and social characteristics, clinical factors, and scores on the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30) were analysed by logistic regression models. A significant proportion (44%) of the patients did not know of their diagnosis of cancer. The scores on functional and symptom scales and global health status/quality of life according to QLQ-C30 did not differ according to the information given or not given on diagnosis. However, the independently significant determinants of good emotional functioning were male gender (P=0.002), low serum alanine transferase levels (P=0.025), good social functioning (P=0.002), and the absence of constipation (P=0.005). In Turkey, it is still common for cancer patients not to be informed of their diagnosis, and there is a great need to improve this situation. Honest disclosure of the truth does not worsen any dimension of quality of life in general or emotional functioning in particular. On the contrary, those with hepatic dysfunction, female gender, poor social functioning and constipation are the ones at increased risk of poor emotional functioning, and these patients may benefit from psychological screening.

Predictors of distant metastasis at presentation in breast cancer: a study also evaluating associations among common biological indicators.

BACKGROUND: To investigate the correlation among some of the commonly used clinical, pathological factors and newer biological indicators, and to identify the independent predictors of distant metastasis at presentation in patients with breast cancer. METHODS: The pathological specimens from 73 patients with breast cancer were retrospectively evaluated by immunohistochemistry. Data on 13 biological indicators; ER, PR, P53, c-erbB-2, PCNA, CEA, Ki-67, Vimentin, Ulex, Nm23, Cathepsin D, Factor VIII, PS2 together with clinical and pathological factors were collected. RESULTS: A number of highly significant correlations were found among the biological indicators studied. By logistic regression analysis, the predictors of distant metastasis at presentation in univariate tests were tumor diameter, number of lymph nodes involved, P53, c-erbB-2 and grade. In multivariate analysis, tumor diameter (P = 0.042, HR: 1.88(1.02-3.44)), c-erbB-2 expression (P=0.035, HR: 18.20 (1.23-268.66)) and grade (P=0.010, HR: 8.05(1.66-39.00)) retained their significance. CONCLUSION: Our findings show that inactivation of suppressor genes, expression of oncogenes, loss of differentiation, augmentation of proliferative activity, metastatic potential, angiogenesis and hormone receptor status are all interrelated facets of breast cancer pathogenesis. Patients with tumors overexpressing c-erbB-2 or with bigger or higher-grade tumors probably need to be more carefully evaluated for the presence of distant metastasis, thus be better staged, at presentation. This may be a new reason to test c-erbB-2 routinely in all patients with breast cancer in addition to its well-known prognostic and predictive uses.

Factors associated with utilization of nonproven cancer therapies in Turkey. A study of 135 patients from a single center.

In their search for a cure, a significant number of cancer patients use nonproven treatment (NPT) methods. However, little is known about patient and disease characteristics associated with the use of these methods. In this trial, we evaluated the prevalence of and the factors associated with the usage of nonproven cancer remedies in a teaching hospital in Turkey. A self-administered questionnaire was given to 135 cancer patients attending the outpatient clinics of a medical oncology department. Patients' demographic data, their usage of nonproven methods, and possible contributing factors were explored. Our cohort mainly comprised poor patients with only primary school education. Overall, 50% of our patients had used or were using NPT methods. Medicinal herbs (mainly stinging nettle) were the most frequently used remedy. In contrast, such "complementary therapies" as exercise, relaxation, and meditation were not employed. In multivariate analysis, only duration of disease was found to be significantly associated with NPT utilization [P=0.05, relative risk (RR)=1.94]. In addition, patient education level was marginally significant (P=0.07, RR=0.36). Apart from long duration of disease and being better educated, no other clinical, social, economic and cultural factors evaluated were associated with the use of NPT in our group of Turkish patients. Since these treatments are sometimes costly and have questionable efficacy and toxicity, proper scientific trials are needed to clarify whether such methods have a real role in cancer management.

Primary lymphoma of bones.

Primary malignant lymphoma of bone (PLB) is an uncommon tumour. A survey of 278 primary malignant cases in our clinic showed that 20 cases of PLB were registered between 1986 and 1997. Fourteen of the 20 cases underwent surgical excision. The mean follow-up time was 36.3 months. The rate of response to treatment was 65% with a rate of complete recovery of 55%. The clinical stage of tumours correlated well with the prognosis. Our results seem worse than most of the series in the literature. A high proportion of stage IV disease and pathological fractures at presentation may be a factor.

Prospective randomized comparison of morning versus night daily single subcutaneous administration of granulocyte-macrophage-colony stimulating factor in patients with soft tissue or bone sarcoma.

BACKGROUND: Hematopoietic growth factors (HGFs) have been used to reduce the neutropenic complications of cytotoxic chemotherapy so that higher doses may be given. The authors have previously shown that endogenous serum granulocyte-colony stimulating factor (G-CSF) and granulocyte-macrophage-colony stimulating factor (GM-CSF) levels at night (p.m.) were significantly higher than those in the morning (a.m.). METHODS: Twenty-four patients with soft tissue or bone sarcoma who were treated with high dose ifosfamide-based chemotherapy were enrolled in this study. Patients were randomized to either a.m. or p.m. treatment. GM-CSF was administered at a dose of 5 microg/kg/day at 10 a.m. or 10 p.m., beginning 36-48 hours after the last chemotherapy dose. GM-CSF therapy was continued until the neutrophil count exceeded 1,000/mm3 for 2 consecutive days. Leukocyte, neutrophil, monocyte, and platelet counts were measured immediately before GM-CSF administration and exactly 12 hours after the first dose of GM-CSF, and every 24 hours until 3 days after the cessation of GM-CSF. RESULTS: The mean duration of Grade 3-4 neutropenia was 5.3 +/- 0.4 days for the a.m. treatment arm and 6.5 +/- 0.3 days for the p.m. treatment arm (P = 0.017). Although the duration of neutropenia in the a.m. arm was significantly shorter than in the p.m. arm, there were no differences related to the number of febrile neutropenic episodes or the duration of antibiotic administration. Also, there were no differences in the side effects observed in the a.m. and p.m. arms. CONCLUSIONS: The finding of 1.2 days' difference in the duration of Grade 3-4 neutropenia warrants further study of chronotherapy with HGFs.