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Supplementary Figure S1-S6 and Supplementary Tables S1-S8

Background: Distinguishing between seizure and neurally mediated syncope is challenging because of similar consequences and medical history. A head-up tilt test (HUTT) is a non-invasive, simple, and easy test to distinguish between epilepsy and syncope besides detailed history taking. Objectives: This study aimed to differentiate between epileptic events and reflex syncope (any different type of syncope) using the head-upright tilt test. Methods: We studied 59 patients (37 boys and 22 girls) between 4 to 18 years old (mean age, 10.5 ± 3.7 years) with a previous diagnosis of seizure who did not respond well to treatment. All patients underwent HUTT, and the test was positive in 26 patients. There were no significant differences in sex, age, provocative factors, associated syndrome, and family history between negative and positive groups. Results: There was a history of actual syncope in 26.9% of the positive tilt test group compared to 15.15% of the negative test group. Also, there was a positive family history of syncope in the positive tilt test group. Among 26 patients with a positive tilt test, 17 were diagnosed with vasovagal syncope (VVS) vasodepressor type and 9 with mixed type. Antiepileptic drugs were tapered for patients diagnosed with VVS, and they did not show any seizures after 18 ± 6 months of follow-up. Overemphasizing positive family history and inattention to history taking are 2 crucial factors leading to the misdiagnosis of epilepsy. Conclusions: Our study showed that HUTT is a non-invasive test that can be useful, especially for early and proper diagnosis in children with refractory epilepsy.

Background Anal squamous cell carcinoma (SCCA) is an uncommon malignancy with a rising incidence that has a high cure rate in its early stages. There is an unmet need for a reliable method to monitor response to treatment and assist in surveillance. Circulating tumor DNA (ctDNA) testing has shown great promise in other solid tumors for monitoring disease progression and detecting relapse in real time. This study aimed to determine the feasibility and use of personalized and tumor-informed ctDNA testing in SCCA. Patients and Methods We analyzed real-world data from 251 patients (817 plasma samples) with stages I-IV SCCA, collected between 11/5/19 and 5/31/22. The tumor genomic landscape and feasibility of ctDNA testing was examined for all patients. The prognostic value of longitudinal ctDNA testing was assessed in patients with clinical follow-up (N = 37). Results Whole-exome sequencing analysis revealed PIK3CA as the most commonly mutated gene, and no associations between mutations and stage. Anytime ctDNA positivity and higher ctDNA levels (MTM/mL) were associated with metastatic disease (P = .004). For 37 patients with clinical follow-up, median follow-up time was 21.0 months (range: 4.1-67.3) post-diagnosis. For patients with stages I-III disease, anytime ctDNA-positivity after definitive treatment was associated with reduced DFS (HR: 28.0; P = .005). Conclusions Our study demonstrates the feasibility of personalized and tumor-informed ctDNA testing as an adjunctive tool in patients with SCCA as well as potential use for detection of molecular/minuteimal residual disease, and relapse during surveillance. Prospective studies are needed to better evaluate the use of ctDNA testing in this indication.

Introduction: Two of the most important tests used in the forensic genetics are DNA fingerprinting and paternity testing. Short tandem repeats (STRs) are very frequently used in identification. Nevertheless, there are limitations on the use of these markers in identifying badly degraded DNA. In these cases, single nucleotide polymorphisms (SNPs) are used because of their shorter amplicon lengths. Recognition of SNPs of high informativeness is a vital step in preparing a list of suitable SNPs. The present research aimed to determine the maximum informative SNPs to be used for identification in Iranian Azari population. Materials and Methods: Four SNPs developed by the SNPforID Consortium were selected. The allele frequencies of the SNPs were obtained using HRM Analysis on DNA samples taken from 100 different individuals. The SNPs that satisfied the Hardy-Weinberg equilibrium and had heterozygosity of higher than 0.50 and equal allele frequency were selected as SNPs of the maximum informativeness. Results: Two polymorphisms (Rs2107612 and Rs1355366) had equal allele frequencies and heterozygosity of higher than 50 percent. Therefore, these 2 polymorphisms are considered highly informative among the studied Azeri population and can be considered in preparing a list of suitable SNPs. Conclusions: Results of the present study can be used along with other SNPs to increase the identification power for some samples. It can also help preparing a database of suitable SNPs to be used for identification in Azeri population in Iran.

The analysis of ancient DNA (aDNA) can inspire both the public and the scientific community. Knowing about ancient human genomes and comparing them with those of modern humans can give us a new perspective on evolution and the migration of humans over time. aDNA is DNA isolated from ancient specimens. It can also be loosely described as any DNA recovered from biological samples that have not been preserved specifically for later DNA analysis. Examples include DNA recovered from archaeological and historical skeletal material, mummified tissues, archival collections of non-frozen medical specimens, preserved plant remains, ice and permafrost cores, Holocene plankton in marine and lake sediments, and so on. Due to considerable anthropological, archaeological, and public interest, human remains receive ample attention from the DNA community. Genetic genealogy is the use of DNA testing in combination with traditional genealogical methods to infer relationships between individuals and to find ancestors. Genetic genealogy involves the use of genealogical DNA testing to determine the level and type of genetic relationship between individuals. DNA markers such as autosomal single nucleotide polymorphisms (SNPs), Y SNPs, and mitochondrial DNA (mtDNA) SNPs are used. By analyzing the sequence of mtDNA and the Y-chromosome, the path of human migration throughout history and the common ancestor of humans can be identified. mtDNA analysis is a field of research in genetics and molecular archaeology that is efficient in less than ideal conditions, such as with biologically degraded materials. The mtDNA molecule not only has a high copy number, but it can also be extracted from very decayed biological specimens. Its D-loop region is polymorphic, consisting of two hypervariable regions (HVI and HVII) with a large variety in different human populations. The analysis of such mtDNA regions using ancient excavated human bones will determine the genetic composition of human mtDNA known as haplogroups and can be used to identify ancient ethnic groups, trace descendants of ancestors, and follow man’s migration trails.

We sought to uncover genetic drivers of hormone receptor–positive (HR+) breast cancer, using a targeted next-generation sequencing approach for detecting expressed gene rearrangements without prior knowledge of the fusion partners. We identified intergenic fusions involving driver genes, including PIK3CA, AKT3, RAF1, and ESR1, in 14% (24/173) of unselected patients with advanced HR+ breast cancer. FISH confirmed the corresponding chromosomal rearrangements in both primary and metastatic tumors. Expression of novel kinase fusions in nontransformed cells deregulates phosphoprotein signaling, cell proliferation, and survival in three-dimensional culture, whereas expression in HR+ breast cancer models modulates estrogen-dependent growth and confers hormonal therapy resistance in vitro and in vivo. Strikingly, shorter overall survival was observed in patients with rearrangement-positive versus rearrangement-negative tumors. Correspondingly, fusions were uncommon ( Significance: By using a powerful clinical molecular diagnostic assay, we identified expressed intergenic fusions as frequent contributors to treatment resistance and poor survival in advanced HR+ breast cancer. The prevalence and biological and prognostic significance of these alterations suggests that their detection may alter clinical management and bring to light new therapeutic opportunities. Cancer Discov; 8(3); 336–53. ©2017 AACR. See related commentary by Natrajan et al., p. 272. See related article by Liu et al., p. 354. This article is highlighted in the In This Issue feature, p. 253

6 Background: For patients with anal squamous cell carcinoma (ASCC), the current standard of care involves curative-intent definitive chemoradiation. For those that recur locally, salvage surgery maybe a consideration. ASCC, however, still lacks a non-invasive blood-based biomarker, which can be of value in this patient population for monitoring recurrence and/or response to immunotherapy later. Circulating tumor DNA (ctDNA) is a promising non-invasive tool to assess molecular residual disease (MRD) and recurrence in ASCC. Here, we evaluated real-world utility of ctDNA status to identify MRD and recurrence in ASCC patients across all stages. Methods: This is a retrospective analysis of patients with any stage ASCC receiving SOC or/and immunotherapy with immune checkpoint inhibitors. A personalized tumor-informed PCR/NGS-based assay (Signatera) was used for ctDNA detection, in the pre/on/post-treatment and surveillance setting. Results: In this study, plasma samples (n=105) were collected from 25 ASCC patients (13 females, 12 males; median age 66 years) at various timepoints for a median follow-up of 315 days (range: 59-1717), post-diagnosis; 12 patients were HPV-positive, 1 patient was HPV-negative, and 12 had unknown HPV status; 88% (22/25) of the patients had serial timepoints (≥2) available. ctDNA-positivity rates, test results, and ctDNA quantification by stage are summarized in Table. The quantitated ctDNA values (mean tumor molecules (MTM)/mL) increased in concordance with the stage of the disease, with values trending higher in stage III and IV. In addition, complete clinical outcome information was available for 22/25 patients at the time of releasing this data. Of the 96 plasma samples drawn from these patients, 68 (70%) were in the surveillance setting (post-definitive therapy). No recurrences were observed among 15 patients who cleared ctDNA on treatment and/or tested negative post-treatment, whereas, 6/7 ctDNA-positive patients were confirmed to have disease recurrence and 1 was pending confirmatory imaging. This often predated recurrence on scans. Conclusions: Measuring and monitoring MRD in patients with ASCC is feasible and has the potential to impact clinical decision making. Our study is the first to set the benchmark for the feasibility of using a tumor-informed assay in ASCC. Larger prospective studies are needed to explore the clinical utility of ctDNA status to guide disease surveillance and management of ASCC.[Table: see text]

Introduction: In psychiatric settings, aggressive events frequently occur during therapy. The use of a proper standard scale to register aggression can facilitate the assessment and control of aggression and help reduce its frequency and severity. The aim of this study is to evaluate the validity and reliability of the Staff Observation Aggression Scale—Revised (SOAS-R). Methods: This psychometric study of the scale was conducted to determine the validity and reliability of the SOAS-R. The validation of the scale was assessed on the basis of 319 aggressive events in the psychiatric wards of the Baqiyatallah and Roozbeh hospitals. Convenience sampling was used for subject selection. Psychometric properties of SOAS-R were studied in two stages. First, the standard scale was translated according to the International Quality of Life Assessment (IQOLA) translation methodology. The face validity, content, and construct validity of the translated version were then determined. The construct validity of the scale was assessed by comparing the known groups. Results: The internal consistency of the whole scale was 0.99. The intra-class correlation coefficients (ICC) were 0.85–0.99 while kappa coefficient was 0.43 to 0.65 for different aspects of the SOAS-R. The validity of the scale was concurrently assessed by using the Visual Analogue Scale (VAS), with a Spearman-Brown correlation coefficient of 0.90. Conclusion: These results showed a favourable validity and reliability for the Persian version of the SOAS-R for the assessment of aggressive behaviour in psychiatric patients.

Background: Thalassemia patients need repeated transfusion that lead to increased blood ferritin level and iron overload in the heart and liver. Because the roles of hepcidin antimicrobial peptide (HAMP) and hemocromatosis protein (HFE) in iron metabolism have been confirmed, this study investigated the effects of these gene's polymorphisms on blood ferritin levels and iron overload in the heart and liver in patients with beta thalassemia major Materials and Methods: This cross-sectional study was conducted on 91 patients referring to the Hajar Hospital in Shahrekord, Iran in 2015. After the blood samples were collected, the ferritin levels were measured, DNA was extracted from the blood cells, and the types of polymorphisms were determined using PCR-RFLP. Data of MRI T2* in the heart and liver were drawn from the patients' medical files. Data analysis was conducted by t-test, chi-square test, Fisher's exact test, and Pearson correlation coefficient. Results: There was no significant correlation between blood ferritin level and c.-582 A>G polymorphisms of hepcidin gene (p=0.58), and H63D of HFE gene (p=0.818). In addition, there was no significant association between the polymorphisms and heart and liver MRI, but there was a significant association between blood ferritin level and qualitative heart and liver MRI (r=-0.34, p=0.035 and r=-0.001, p=0.609, respectively). Conclusion: In patients with β-thalassemia major, the presence of c.-582A>G HAMP and H63D HFE polymorphisms is not effective on blood ferritin level and iron overload in the heart and liver in the studied region.

539 Background: Male BC accounts for < 1% of all BC and is often diagnosed at later stage, which can result in higher mortality. Due to the rarity of this diagnosis, limited data exist on genomic alterations and prevalence of cancer susceptibility genes (CSG). We aimed to comprehensively describe the genomics of male BC and compare these to a FBC cohort across subtypes to provide insight on tumor biology and opportunities for targeted therapies. Methods: 275 male BC TBx or LBx were sequenced by Foundation Medicine (FM) using hybrid capture-based CGP. Both TBx and LBx were evaluated for all classes of genomic alterations (GA). Histological subtype, receptor status, and biopsy site were extracted from pathology reports. Paired samples with both TBx and LBx were available in 7 cases. The male BC TBx cohort with known receptor status (n = 253) was compared to a FBC cohort (n = 2855). Mutational prevalence in 5 breast CSG (ATM/BRCA1/BRCA2/CHEK2/PALB2) were compared along with their associated genomic LOH (gLOH) values. Results: Among male BCs, subtype distribution was: ER+/HER2- n = 210 (83%), ER+/HER2+ n = 22 (9%), TNBC n = 20 (8%). ER+/HER2+ male BC cases had higher rates of ERBB2 SV (22.7% v. 0.62%, p < 0.0001), PIK3CA (68.2% vs. 34%, p = 0.01), MDM2 amplifications (36% v. 4%, p < 0.0001) and GATA3 (36.6% v. 6.2%, p = 0.0002) than ER+/HER2+ FBC. In the ER+/HER2- cohort, male BC had more alterations in BRCA2 (13.8% v. 5.3%, p < 0.0001) and GATA3 (26% vs. 15%, p = 0.0004) and less alterations in TP53 and ESR1 (p < 0.0001 both). 28.6% of male BC v. 16.6% of FBC (p = 0.004) had one or more variants in one of 5 CSG of potential germline origin with a higher % of BRCA mutations in male BC vs. FBC (17.5% v. 9.9%, p = 0.0006). In the paired male BC Bx’s we saw genomic heterogeneity in a case showing 5 unique ESR1 alts and a PIK3CA SV unique to LBx done at the same time as TBx. We saw evidence of resistance with a shared BRCA1 alteration and several reversion mutations unique to LBx taken 471 days later, and a longitudinal pair with unique ESR1,PIK3CA and MTOR mutations in a LBx 547 days apart. Conclusions: Although male and female BC share some common alterations, our study revealed potentially important findings that may explain biological differences and provide treatment opportunities. Despite HR+/HER2+ male BC being rare, it was notable for increased co-mutations with ERBB2 SV, PIK3CA SV, and GATA3 SV, which can be associated with a worse prognosis but perhaps allow more novel combinations. ESR1 mutations appear more common in ER+/HER2- FBC reflective of treatment with aromatase inhibitors versus tamoxifen for male BC. TP53 mutations were more common in all subtypes of FBC. BRCA2 mutations and other potential germline CSG variants were more common in male BC, suggesting an opportunity for PARP inhibitors. LBx identified additional biomarkers and resistance mutations not seen in TBx.

Aim: Nowadays, life is endangering women’s health, so that without organizational support, women are engaged in unhealthy life styles. Osteoporosis is the most common metabolic bone disease that increases the risk of bone fracture by creating a structural abnormality in the bone. Osteoporosis is a common disease among middle-age and older persons, especially women. Therefore, the necessity of implementing strategic plans to prevent osteoporosis is significantly important. This study was designed to determine the prediction of osteoporosis preventive behaviors using the Health Belief Model (HBM).Methods: This cross-sectional study was done among 200 women referred to the health centers in Taft in 2016. A stage simple random sampling was used. In order to gather the data a questionnaire consistent with the structures of HBM was used. Data were analyzed using the SPSS v16 and descriptive statistics (frequency, percentage, and mean±SD) and analysis (Linear regression). The significance level was 0.5.Findings: The mean age of the participants was 53 years. Linear regression analysis showed the perceived awareness among the participants (p> 0.001).Conclusion: The results showed that HBM is able to predict well the osteoporosis preventive behaviors in women. The findings of this survey confirm the efficiency of HBM in adopting preventive actions of osteoporosis.

// Laurence Booth 1 , Thomas Albers 3 , Jane L. Roberts 1 , Mehrad Tavallai 1 , Andrew Poklepovic 2 , Iryna O. Lebedyeva 3 , Paul Dent 1 1 Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA 23298, USA 2 Department of Medicine, Virginia Commonwealth University, Richmond, VA 23298, USA 3 Department of Chemistry and Physics, Augusta University, Summerville Campus, Augusta GA 30912, USA Correspondence to: Paul Dent, email: pdent@vcu.edu Keywords: sorafenib, pazopanib, chaperones, ERBB, PI3K Received: April 25, 2016 Accepted: May 20, 2016 Published: May 31, 2016 ABSTRACT We have recently demonstrated that multi-kinase inhibitors such as sorafenib and pazopanib can suppress the detection of chaperones by in situ immuno-fluorescence, which is further enhanced by phosphodiesterase 5 inhibitors. Sorafenib and pazopanib inhibited the HSP90 ATPase activity with IC50 values of ~1.0 μM and ~75 nM, respectively. Pazopanib docked in silico with two possible poses into the HSP90 ATP binding pocket. Pazopanib and sildenafil combined to reduce the total protein levels of HSP1H/p105 and c-MYC and to reduce their co-localization. Sorafenib/pazopanib combined with sildenafil in a [GRP78+HSP27] –dependent fashion to: (i) profoundly activate an eIF2α/Beclin1 pathway; (ii) profoundly inactivate mTOR and increase ATG13 phosphorylation, collectively resulting in the formation of toxic autophagosomes. In a fresh PDX isolate of NSCLC combined knock down of [ERBB1+ERBB3] or use of the ERBB1/2/4 inhibitor afatinib altered cell morphology, enhanced ATG13 phosphorylation, inactivated NFκB, and further enhanced [sorafenib/pazopanib + sildenafil] lethality. Identical data to that with afatinib were obtained knocking down PI3K p110α/β or using buparlisib, copanlisib or the specific p110α inhibitor BYL719. Afatinib adapted NSCLC clones were resistant to buparlisib or copanlisib but were more sensitive than control clones to [sorafenib + sildenafil] or [pazopanib + sildenafil]. Lapatinib significantly enhanced the anti-tumor effect of [regorafenib + sildenafil] in vivo ; afatinib and BYL719 enhanced the anti-tumor effects of [sorafenib + sildenafil] and [pazopanib] in vivo , respectively.

// Mehrad Tavallai 1 , Laurence Booth 1 , Jane L. Roberts 1 , William P. McGuire 2 , Andrew Poklepovic 2 and Paul Dent 1 1 Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA 2 Department of Medicine, Virginia Commonwealth University, Richmond, VA, USA Correspondence to: Paul Dent, email: // Keywords : ruxolitinib, JAK-STAT, DMF, tecfidera, ROS Received : January 28, 2016 Accepted : February 29, 2016 Published : March 15, 2016 Abstract We determined whether the myelofibrosis drug ruxolitinib, an inhibitor of Janus kinases 1/2 (JAK1 and JAK2), could interact with the multiple sclerosis drug dimethyl-fumarate (DMF) to kill tumor cells; studies used the in vivo active form of the drug, mono-methyl fumarate (MMF). Ruxolitinib interacted with MMF to kill brain, breast, lung and ovarian cancer cells, and enhanced the lethality of standard of care therapies such as paclitaxel and temozolomide. MMF also interacted with other FDA approved drugs to kill tumor cells including Celebrex ® and Gilenya ® . The combination of [ruxolitinib + MMF] inactivated ERK1/2, AKT, STAT3 and STAT5; reduced expression of MCL-1, BCL-XL, SOD2 and TRX; increased BIM expression; decreased BAD S112 S136 phosphorylation; and enhanced pro-caspase 3 cleavage. Expression of activated forms of STAT3, MEK1 or AKT each significantly reduced drug combination lethality; prevented BAD S112 S136 dephosphorylation and decreased BIM expression; and preserved TRX, SOD2, MCL-1 and BCL-XL expression. The drug combination increased the levels of reactive oxygen species in cells, and over-expression of TRX or SOD2 prevented drug combination tumor cell killing. Over-expression of BCL-XL or knock down of BAX, BIM, BAD or apoptosis inducing factor (AIF) protected tumor cells. The drug combination increased AIF : HSP70 co-localization in the cytosol but this event did not prevent AIF : eIF3A association in the nucleus.

11029 Background: In recent years, genomic profiling has become standard of care for several gastrointestinal (GI) cancers. In addition to standard of care indications, comprehensive genomic profiling has led to novel and expanded applications of targeted therapy, chemotherapy, and immunotherapy and facilitated identification of potential clinical trials. A GI molecular tumor board (MTB) was developed with a goal of improving understanding of the biological effects of genomic alterations and their therapeutic implications to enhance personalized therapy. Methods: Foundation Medicine (FM) collaborated with physicians in the GI oncology group of an academic medical center to develop a GI MTB starting March 2019. As of December 2019, 27 GI oncology cases were presented where FoundationOneCDx testing was performed and a clinical question was posed. Cases were discussed by faculty, fellows, research staff, and a clinical genomic scientist and oncologist from FM. Impacted signaling pathways and biomarkers were discussed for each case alongside clinical content so that physicians could consider therapeutic options and clinical trials. Presenting faculty were asked to complete a questionnaire for each case presented to assess the impact of the MTB discussion on clinician knowledge and patient-level treatment recommendations. Results: Of 27 questionnaires sent to 7 providers, 17 (63%) were completed. Respondents indicated that as a result of the MTB, the treatment plan was changed in 2 cases (12%), reinforced in 9 cases (53%) and in 6 cases (35%) there was no effect. On a Likert scale of 1-4 where 1 is “rare/poorly” and 4 is “great” mean scores were as follows: Did this MTB help you understand the biological effects of the main genomic alteration(s) reported in the case presented? 3.3. Did this MTB help you understand the possible therapeutic implications of the main genomic alterations in the case presented? 3.3. Did this MTB improve your understanding of the role of next generation sequencing and comprehensive genomic profiling in making treatment decisions? 3.4. Conclusions: The results of our questionnaire indicate that treatment decisions were changed in a minority of cases based on the MTB. In most cases, clinical decision making was reinforced and understanding of the biological effects of genomic alterations and their therapeutic implications were improved. Based on this feedback we will continue to refine and integrate the GI MTB into clinical care for patients with GI malignancies, and share our experience locally with other disease groups.

Background: Thalassemia patients need repeated transfusion that lead to increased blood ferritin level and iron overload in the heart and liver. Because the roles of hepcidin antimicrobial peptide (HAMP) and hemocromatosis protein (HFE) in iron metabolism have been confirmed, this study investigated the effects of these gene's polymorphisms on blood ferritin levels and iron overload in the heart and liver in patients with beta thalassemia major Materials and Methods: This cross-sectional study was conducted on 91 patients referring to the Hajar Hospital in Shahrekord, Iran in 2015. After the blood samples were collected, the ferritin levels were measured, DNA was extracted from the blood cells, and the types of polymorphisms were determined using PCR-RFLP. Data of MRI T2* in the heart and liver were drawn from the patients' medical files. Data analysis was conducted by t-test, chi-square test, Fisher's exact test, and Pearson correlation coefficient. Results: There was no significant correlation between blood ferritin level and c.-582 A>G polymorphisms of hepcidin gene (p=0.58), and H63D of HFE gene (p=0.818). In addition, there was no significant association between the polymorphisms and heart and liver MRI, but there was a significant association between blood ferritin level and qualitative heart and liver MRI (r=-0.34, p=0.035 and r=-0.001, p=0.609, respectively). Conclusion: In patients with β-thalassemia major, the presence of c.-582A>G HAMP and H63D HFE polymorphisms is not effective on blood ferritin level and iron overload in the heart and liver in the studied region.

Prior tumor cell studies have shown that the drugs sorafenib (Nexavar) and regorafenib (Stivarga) reduce expression of the chaperone GRP78. Sorafenib/regorafenib and the multi‐kinase inhibitor pazopanib (Votrient) interacted with sildenafil (Viagra) to further rapidly reduce GRP78 levels in eukaryotes and as single agents to reduce Dna K levels in prokaryotes. Similar data were obtained in tumor cells in vitro and in drug‐treated mice for: HSP70, mitochondrial HSP70, HSP60, HSP56, HSP40, HSP10, and cyclophilin A. Prolonged ‘rafenib/sildenafil treatment killed tumor cells and also rapidly decreased the expression of: the drug efflux pumps ABCB1 and ABCG2; and NPC1 and NTCP, receptors for Ebola/Hepatitis A and B viruses, respectively. Pre‐treatment with the ‘Rafenib/sildenafil combination reduced expression of the Coxsackie and Adenovirus receptor in parallel with it also reducing the ability of a serotype 5 Adenovirus or Coxsackie virus B4 to infect and to reproduce. Sorafenib/pazopanib and sildenafil was much more potent than sorafenib/pazopanib as single agents at preventing Adenovirus, Mumps, Chikungunya, Dengue, Rabies, West Nile, Yellow Fever, and Enterovirus 71 infection and reproduction. ‘Rafenib drugs/pazopanib as single agents killed laboratory generated antibiotic resistant E. coli which was associated with reduced Dna K and Rec A expression. Marginally toxic doses of ‘Rafenib drugs/pazopanib restored antibiotic sensitivity in pan‐antibiotic resistant bacteria including multiple strains of bla kpc Klebsiella pneumoniae. Thus, Dna K is an antibiotic target for sorafenib, and inhibition of GRP78/Dna K has therapeutic utility for cancer and for bacterial and viral infections. J. Cell. Physiol. 230: 2552–2578, 2015. © 2015 The Authors. Journal of Cellular Physiology published by Wiley Periodicals, Inc.

Pancreatic cancer has the lowest 5-year survival rate of all major cancers despite decades of effort to design and implement novel, more effective treatment options. In this study, we tested whether the dual phosphoinositide 3-kinase/mechanistic target of rapamycin inhibitor BEZ235 (BEZ) potentiates the antitumor effects of doxorubicin (DOX) against pancreatic cancer. Cotreatment of BEZ235 with DOX resulted in dose-dependent inhibition of the phosphoinositide 3-kinase/mechanistic target of rapamycin survival pathway, which corresponded with an increase in poly ADP ribose polymerase cleavage. Moreover, BEZ cotreatment significantly improved the effects of DOX toward both cell viability and cell death in part through reduced Bcl-2 expression and increased expression of the shorter, more cytotoxic forms of BIM. BEZ also facilitated intracellular accumulation of DOX, which led to enhanced DNA damage and reactive oxygen species generation. Furthermore, BEZ in combination with gemcitabine reduced MiaPaca2 cell proliferation but failed to increase reactive oxygen species generation or BIM expression, resulting in reduced necrosis and apoptosis. Treatment with BEZ and DOX in mice bearing tumor xenographs significantly repressed tumor growth as compared with BEZ, DOX, or gemcitabine. Additionally, in contrast to the enhanced expression seen in MiaPaca2 cells, BEZ and DOX cotreatment reduced BIM expression in H9C2 cardiomyocytes. Also, the Bcl-2/Bax ratio was increased, which was associated with a reduction in cell death. In vivo echocardiography showed decreased cardiac function with DOX treatment, which was not improved by combination treatment with BEZ. Thus, we propose that combining BEZ with DOX would be a better option for patients than current standard of care by providing a more effective tumor response without the associated increase in toxicity.