Term Side Effects Research Articles

SM88/SMK non-hormonal therapy in recurrent or untreated prostate cancer.

e16540 Background: Prostate cancer (PC) has a poor prognosis. Androgen deprivation therapy is effective but has unacceptable short and long term side effects. SM88 (modulators of mTOR, mitochondrial stress, CYP3a4 and tyrosine isomers) is a novel combination of anti-cancer therapies with previously reported favorable results yet low toxicity in a heterogeneous group of 30 patients with cancer. We now report on a PC cohort treated with SM88. Methods: Retrospective chart review. Results: Between 2012 and 2014, 6 men with PC were treated with SM88, administered 5 days/week over a 6-week treatment cycle. SM88 consisted of daily oral and subcutaneous injection of the components. The average age was 54 years (range 32-66). Five (83.3%) were Caucasian, 1 (16.7%) was Hispanic. patients with prostate cancer were treated with SM88 daily, Monday-Friday, for between 6 to 31 wks (median 11). Four patients had castrate resistant disease, two declined ADT. One had failed prior chemotherapy (see Table). All subjects were not receiving other therapy. One subject died of disease complications approximately 3 months after starting treatment but only received approximately 20 days of therapy over a 6 week cycle. Three patients had PSA nadir responses of >90% reduction over pretreatment peak levels (see table). Two others had radiographic SD for between 10-14 wks. There were no significant toxicity except cutaneous hyperpigmentation. The duration of responses ranged from 10-114 weeks. Best overall responses included 2 patients with complete biochemical response. Four subjects improved their ECOG score during treatment and 2 had no change. Conclusions: SM-88 appears to be well tolerated and to have anti-tumor activity in prostate cancer without androgen deprivation toxicity. Additional confirmatory prospective studies in prostate are ongoing and planned in other cancers. [Table: see text]

Feasibility intervention to increase exercise outcome expectations among breast cancer survivors.

e21584 Background: Exercise is associated with decreased mortality and recurrence risk and improved quality of life for breast cancer survivors (BCS). Roughly 1/3 of BCS exercise enough to experience these benefits. A promising strategy to increase exercise among BCS is to target outcome expectations (OEs), or what one expects to obtain or avoid from a behavior. OE dimensions include 1) importance – value placed on the outcome(s); 2) certainty - perceived probability outcome(s) will occur; and 3) accessibility – the frequency with which outcome(s) are considered. Methods: This RCT tested the feasbility and effects of a self-directed booklet designed to increase OEs and exercise. It was created in collaboration with BCS who successfully use exercise to manage long term side effects. OEs were measured through online surveys and exercise was measure with a Fitbit accelerometer. Data was collected at baseline, 4, 8 and 12 weeks post intervention. Feasibility was defined as the extent to which participants made use of intervention materials and was assessed through 9 researcher related questions. Results: The sample is mean age of 58, 74% white, 26% black, an average 2.5 years post treatment and about 1/3 take an AI or SERM. The booklet “somewhat” (mean = 3.24, SD = 1.8 on 1-5 Likert scale) made use of the booklet, as intended. OEs increased 0.01 points over time in the intervention arm, compared to the control arm ( p = 0.3555); 0.4 points more for black participants compared to white ( p = 0.0003); 0.2 points per every unit of increased exercise self-efficacy ( p < 0.0001), and 0.01 points per month participants were closer to their time of cancer surgery ( p = 0.0037). Intervention arm participants increased their weekly steps by 970, every 4 weeks, compared to participants in the control arm ( p = 0.0283). Conclusions: Overall the intervention was well received by participants, had positive effects on OEs and exercise and can easily be translated into practice with need for minimal resources. Significant differences were noted for black compared to white participants, thus it is necessary to understand racial differences in exercise beliefs when designing future exercise interventions. Clinical trial information: NCT02348710.

IgG4-related membranous glomerulonephritis and generalized lymphadenopathy without pancreatitis: a case report

BackgroundIgG4-related disease is a recently described pathologic entity. This is the case of a patient with nephrotic syndrome and lymphadenopathy due to IgG4-related disease. Such a kidney involvement is quite peculiar and has only been described a few times recently. Renal biopsy showed a glomerular involvement with membranous glomerulonephritis in association with a tubulo-interstitial nephropathy. Moreover, the patient was not suffering from pancreatitis.Case presentationThe patient is a middle-aged man of Moroccan origin. He has developed recurrent episodes of diffuse lymphadenopathies, renal failure and nephrotic syndrome. Renal biopsies showed membranous glomerulonephritis.Discussion and conclusionThe diagnostic approach of this atypical presentation is discussed in this case report as well as diagnostic criteria, therapeutic strategies, biomarkers and pathophysiology of IgG4-related disease. IgG4-related membranous glomerulonephritis is a well-established cause of membranous glomerulonephritis. It must be sought after in every patient with a previous diagnosis of IgG4-related disease and in every patient with this histological finding on renal biopsy. Corticoids are still the first-line treatment of IgG4-related disease. New therapeutic strategies are needed to avoid glucocorticoids long term side-effects. Interestingly, the patient was prescribed cyclophosphamide in addition to glucocorticoids for an immune thrombocytopenia. This treatment had a very good impact on his IgG4-related disease.

40P - Antitumoral effect and reduced systemic toxicity in mice after intra-tumoral injection of an in vivo solidifying calcium sulfate formulation with docetaxel

Background: Docetaxel is a cytostatic agent approved for treatment of non-small cell lung cancer as well as other cancers. Its effectiveness in clinical practice is associated with a variety of acute and long term side effects. To reduce the systemic side effects, a local slow-release depot formulation based on calcium sulfate with docetaxel for intra-tumoral administration was developed. Methods: Two formulations with a twofold difference in docetaxel drug load (corresponding to 12.5 and 25.0 mg/kg body weight) were manufactured with the calcium sulfate based NanoZolid™ technology. The formulations were injected intra-tumorally as a paste which solidified within the tumor. The effects of the two local depot formulations were tested in female mice (n = 60) inoculated with subcutaneous Lewis lung carcinoma cells. The slow-release depots were compared to systemic intraperitoneal injections of docetaxel (25.0 mg/kg) and a calcium sulfate placebo formulation. Tumor volumes were measured and systemic side effects were evaluated using body weight and cell counts from whole blood as well as plasma concentrations. In addition, a histological evaluation was performed. Results: Both docetaxel formulations showed a significantly higher antitumor efficacy compared to placebo, which was comparable to that of systemic administration of docetaxel. Moreover, the intra-tumoral depot formulations with docetaxel showed reduced systemic toxicity compared to systemic treatment, including less weight loss and no decrease in blood cell counts. The extracellular deposits of calcium sulfate, observed in all groups with intra-tumoral injections, did not lead to significant inflammatory changes. Conclusions: It was shown that intra-tumoral slow-release depots of calcium sulfate with docetaxel were well tolerated and released docetaxel at amounts sufficient for satisfactory local antitumor effect and for a low level plasma detection. The use of such depots can be an alternative to intravenous injection as an efficient antitumoral treatment with reduced systemic toxicity. Legal entity responsible for the study: Uppsala University, Department of Immunology, Genetics and Pathology, Marie Jeansson Funding: This study was supported by an IGP Young Investigator grant (MJ), the Swedish Research Council (521-2012-865, MJ), Åke Wiberg Foundation (738866289, MJ), Magnus Bergvalls Foundation (24942-1-2013, 2014-00055, MJ), and Ture Stenholms Foundation for Surgical Research (MS) Disclosure: S. Grudén: Consultant for LIDDS AB. N. Axén: Consultant for LIDDS AB and have equity interests in LIDDS AB. All other authors have declared no conflicts of interest.

Melatonin as an anti-inflammatory agent in radiotherapy.

Radiotherapy is one of the most relevant treatment options for cancer therapy with or without other treatment modalities including immunotherapy, surgery and chemotherapy. Exposure to heavy doses of ionizing radiation during radiotherapy results in short and long term side effects. It appears that many of these side effects are linked to inflammatory responses during treatment or after prolonged use. Inflammation is mediated by various genes and cytokines related to immune system responses caused by massive cell death following radiotherapy. This phenomenon is more obvious, particularly after exposure to clinical doses of radiotherapy. Inflammation is involved in the amplification of acute responses, genomic instability and also long term pathological changes in normal tissues. Moreover, inflammation attenuates responses of the tumor to radiotherapy through some mechanisms such as angiogenesis. Thus, the management of inflammation is one of the most interesting aims in cancer radiotherapy. Melatonin, known as a natural product in the body, has been of much interest for its anti-inflammatory properties. Some studies have proposed melatonin as a novel anti-inflammation agent. This literature review will concentrate on the anti-inflammatory properties of melatonin that may help the management of different inflammatory signaling pathways in both tumor and normal tissues.

Abstract P5-13-17: Simplifying survivorship surveillance

Abstract Breast cancer survivors represent 4 of 10 female cancer survivors in the United States, comprising an estimated 3.1 million women living with breast cancer in the US. Addressing survivors' unique post-treatment needs is critical to providing quality health care. In fact, Survivorship Care Plans are now required for all breast cancer patients by the National Accreditation Program for Breast Centers. To ensure that we meet recommended guidelines and provide comprehensive survivorship care, we have created a computerized treatment summary and developed a unique simple assessment tool to be incorporated at the patient's survivorship visit. The goal is to increase awareness of short and long term side effects post treatment and develop a plan to address the patient's needs. The visits are carried out by the nurse practitioners and average 60 minutes. Data from the first 20 patients in our new program show that most are age 40-49 (30%), 25% are age 60-69, 20% age 50-59, 20% age 70-79 and 5% age 30-39. The majority of patients (65%) were treated with breast conserving surgery and sentinel node biopsy. Most women who had a mastectomy also had reconstruction (83%). The majority (95%) had lymph nodes removed as part of their cancer surgery and 30% report a sensation of tightness or swelling on the side where lymph nodes were resected. The majority (90%) are receiving hormonal therapy and 40% also had chemotherapy. A majority (75%) reported side effects from hormonal therapy such as hot flashes, joint pain, or vaginal dryness. 15% of patients reported concerns with sexual function or sexual activity. Over a third of patients (35%) felt anxious, nervous, or on edge, 25% of patients reported that they had little interest or pleasure in doing things, 15% felt down, depressed or hopeless. Memory problems were reported by 30% of patients, 25% complained of difficulties multitasking or paying attention and 10% reported their thinking seemed slow. Over half of the patients (55%) reported problems with insomnia, 30% complained that fatigue interfered with their usual activities, 15% had persistent fatigue, and 15% reported excessive sleepiness. 25% complained of ongoing discomfort related to their cancer treatment but only 5% felt the pain interfered with their usual activities. There were no reports of difficulties with activities of daily living. However, 25% had concerns about their body image and appearance since cancer treatment. Most patients (90%) report eating a well-balanced diet and 65% engage in regular physical activity yet 45% of patients are concerned about their weight. Only 5% are smokers, 40% have up to 3 alcoholic beverages a week, 20% have 4-7 drinks a week, 10% drink more than 7 alcoholic beverages a week, and 25% do not drink alcohol at all. Only 50 percent had a recent pap smear, 45% had a recent colonoscopy, 60% had a recent bone density, and 80% see an ophthalmologist regularly. Most patients (60%) did not need referrals for other services, but 5% were sent for mental health services, 10% for consultation with the plastic surgeon, 10% for consultation regarding sexual functioning, 5% for physical therapy, and 5% for social work intervention. We have had an overwhelmingly positive response from patients to the survivorship visits and will continue to expand this useful and effective program. Citation Format: Pories SE, Myers LE, Dusenbery KL. Simplifying survivorship surveillance [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P5-13-17.

Growing a cancer survivorship care plan program.

58 Background: More than 15.5 million cancer survivors live in the United States. This number is expected to be over 20 million by 2026. Cancer survivors have increased risk of morbidity; therefore, preventive and on-going medical treatment requires close monitoring and coordination. The Institute of Medicine’s (IOM) 2005 report, Cancer Patient to Cancer Survivor: Lost in Transition, recommended health providers raise awareness of cancer survivors’ needs and establish cancer survivorship as a distinct phase of care. The IOM also recommended patients who complete primary treatment are provided a comprehensive summary and plan that is effectively explained. A survivorship care plan maps out and improves care related to accessibility of past diagnosis and treatment history, surveillance guidelines, and potential long term side effects. In 2012, the Commission on Cancer (CoC) added Standard 3.3 Survivorship Care Planto the program standards. This met the IOM’s objective of addressing potential patients that get “lost” as they transition from care they received during treatment through phases of their life or disease. Methods: The Cancer Committee within a CoC certified organization developed multiple strategies to address the IOM and CoC standards. Strategies included a process to disseminate a comprehensive care summary for cancer patients who are completing primary treatment, adoption of the American Society of Clinical Oncology’s Treatment Summary and Survivorship Care Plan template, and adding a survivorship nurse navigator to the interprofessional treatment team. The survivorship nurse navigator monitors and reviews survivorship care plans with patients, advises when to seek treatment for symptoms, discusses surveillance guidelines, navigates patients through therapies, and educates on prevention and screening. Results: Evaluation for quality of life and compliance with individualized surveillance guidelines is ongoing. Conclusions: The oncology nurse navigator role is uniquely positioned to lead care coordination and improve outcomes through the continuum of care. Providing patients with a summary of their treatment and a plan moving forward may decrease stress related to the transition from active treatment to survivorship.

An evaluation of survivorship care plan program.

62 Background: The Commission on Cancer mandated the development and implementation of survivorship care plans (SCP) in 2015. Minimal evidence exists to suggest SCP are meaningful to survivors and primary care physicians (PCP). We sought to evaluate the usefulness of the SCP as perceived by both survivors and PCP. Methods: Phase I was 8-month pilot using a 7-page SCP detailing medical information and follow up sections addressed to the survivor and to the PCP. To evaluate patient and PCP views on the SCP, a telephone survey was created for patients and a two-page Likert scale survey for PCP was faxed. Phase I evaluations revealed 90% of survivors did not look past page 2 of the document and 16% of PCP felt it was too long and busy to read. The SCP was edited for a 3-page version and implemented in Phase II over 11-months. Results: In Phase I, 78 SCP were delivered to survivors, 58% were surveyed. Of PCP, 24% responded. In Phase II, 895 SCP were delivered, and 274 (31%) survivors surveyed. In Phase I and II, when asked to identify themselves as a cancer survivor, 282 (88%) confirmed and 21 (7%) were unsure. After reviewing the SCP, 93.1% understood their plan of care; 79.6% understood potential late effects; 33% were aware of support or resources available; 96% would recommend to another cancer survivor to get one. In Phase II, 16% (43) of PCP responded. Overall replies were positive: 61% found resource list helpful; 66% wanted more specific information about patient potential late effects; 87% agreed knowing symptoms of late effects is helpful. 70% wanted clarity on follow up tests needed and who should order. PCP overwhelmingly agreed (88%) the SCP is helpful. 90% agreed that knowing ongoing side effects of treatment was important. Conclusions: Survivors liked having their cancer treatment in one concise document and PCP physicians feel that the SCP is meeting their basic needs. PCP are interested in more specific long term side effects to monitor in their patients and direction on what follow-up tests are needed and who is ordering them. Further research is needed to study the impact of SCP on survivors’ health.

Antitumoral effect and reduced systemic toxicity in mice after intra-tumoral injection of an in vivo solidifying calcium sulfate formulation with docetaxel

BackgroundDocetaxel is a cytostatic agent approved for treatment of non-small cell lung cancer as well as other cancers. Although docetaxel is an effective cytostatic agent, its effectiveness in clinical practice is associated with a variety of acute and long term side-effects. To overcome systemic side-effects, a slow release formulation based on calcium sulfate with docetaxel for intra-tumoral administration was developed. MethodsTwo formulations with the calcium sulfate NanoZolid technology were generated with a twofold difference in docetaxel drug load. The formulations were injected intra-tumorally as a paste which solidified within the tumor. The effects of the two intra-tumoral injection formulations were tested in female mice (n=60) inoculated with subcutaneous Lewis lung carcinoma cells. The two formulations were compared to systemic intraperitoneal injection of docetaxel and a placebo formulation without docetaxel. Tumor volumes were measured and systemic side-effects were evaluated using body weight and cell counts from whole blood as well as plasma concentrations. ResultsBoth docetaxel formulations showed a significantly higher antitumor efficacy compared to placebo, which was comparable to that of systemic administration of docetaxel. Moreover, the intra-tumoral formulations with docetaxel showed reduced systemic toxicity compared to systemic treatment, including less weight loss and no decrease in blood cell counts. ConclusionsThe results suggest that intra-tumoral slow release calcium sulfate based formulations with docetaxel can be an alternative strategy as an efficient local antitumoral treatment with reduced systemic toxicity.

Abstract A25: Systems biology approach to delineating DNA damage response and R-loop dependencies in Ewing sarcoma

Abstract Ewing sarcoma is a rare, yet highly aggressive family of bone and soft tissue tumors that afflicts children and young adults. It is the prototypical example of mesenchymal tumors driven by a fusion oncogene involving the EWSR1 gene, most frequently EWS-FLI1. Ewing sarcoma is exquisitely sensitive to genotoxic agents, and more recently PARP1 inhibitors, predominantly when detected at an early stage. However, the molecular basis for this sensitivity is relatively under-explored. Further, the long term side effects of these drugs pose a critical barrier to effective treatment. In this study, we proposed to delineate the pathways dictating Ewing sarcoma chemosensitivity and identify alternative avenues for targeted therapy. R-loops are three nucleic acid DNA structures consisting of an RNA:DNA hybrid and a displaced complementary DNA loop. The importance of R-loops in both being causative as well as disruptive in DNA damage as well as DNA damage response has emerged through recent studies. Our previous work has demonstrated that Ewing sarcoma cell lines have high levels of endogenous damage as well as replication stress. We observe that Ewing cell lines have elevated levels of persistent R-loops and impaired homologous recombination repair of double strand breaks. In the present study, we use a multimodal, systems biology approach involving high-throughput RNAi based survival data, time-course RNA-seq in response to two chemotherapeutics, Chromatin immunoprecipitation-seq and DNA:RNA hybrid immunoprecipitation-seq datasets across multiple Ewing sarcoma and control cell lines. Our data led to the identification of a subset of genes whose expression patterns in response to DNA damage is dysregulated in Ewing sarcoma. Predominantly, among these genes is a network of RNA binding proteins that highlights the importance of RNA metabolism in DNA damage response. We also observe a concomitant alteration in damage-dependent transcription. Finally, we identified several compensatory mechanisms that are upregulated in Ewing sarcoma (RNASEH2, FEN1, Fanconi Anemia, etc.) in order to manage the exacerbated genomic stress conditions and targeting any of these pathways leads to severe tumor cell cytotoxicity. We therefore propose the use of these pathways as potential candidates for targeted therapy. Citation Format: Aparna Gorthi, Eva Loranc, Cao Lin, Alexander JR Bishop. Systems biology approach to delineating DNA damage response and R-loop dependencies in Ewing sarcoma. [abstract]. In: Proceedings of the AACR Special Conference on Engineering and Physical Sciences in Oncology; 2016 Jun 25-28; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2017;77(2 Suppl):Abstract nr A25.

TDCS for Memory Enhancement: Analysis of the Speculative Aspects of Ethical Issues.

Transcranial direct current stimulation (tDCS) is a promising technology to enhance cognitive and physical performance. One of the major areas of interest is the enhancement of memory function in healthy individuals. The early arrival of tDCS on the market for lifestyle uses and cognitive enhancement purposes lead to the voicing of some important ethical concerns, especially because, to date, there are no official guidelines or evaluation procedures to tackle these issues. The aim of this article is to review ethical issues related to uses of tDCS for memory enhancement found in the ethics and neuroscience literature and to evaluate how realistic and scientifically well-founded these concerns are? In order to evaluate how plausible or speculative each issue is, we applied the methodological framework described by Racine et al. (2014) for “informed and reflective” speculation in bioethics. This framework could be succinctly presented as requiring: (1) the explicit acknowledgment of factual assumptions and identification of the value attributed to them; (2) the validation of these assumptions with interdisciplinary literature; and (3) the adoption of a broad perspective to support more comprehensive reflection on normative issues. We identified four major considerations associated with the development of tDCS for memory enhancement: safety, autonomy, justice and authenticity. In order to assess the seriousness and likelihood of harm related to each of these concerns, we analyzed the assumptions underlying the ethical issues, and the level of evidence for each of them. We identified seven distinct assumptions: prevalence, social acceptance, efficacy, ideological stance (bioconservative vs. libertarian), potential for misuse, long term side effects, and the delivery of complete and clear information. We conclude that ethical discussion about memory enhancement via tDCS sometimes involves undue speculation, and closer attention to scientific and social facts would bring a more nuanced analysis. At this time, the most realistic concerns are related to safety and violation of users’ autonomy by a breach of informed consent, as potential immediate and long-term health risks to private users remain unknown or not well defined. Clear and complete information about these risks must be provided to research participants and consumers of tDCS products or related services. Broader public education initiatives and warnings would also be worthwhile to reach those who are constructing their own tDCS devices.

Ketogenic diet therapy for epilepsy during pregnancy: A case series

PurposeEvaluation of ketogenic diet (KD) therapies for seizure control during pregnancy when safety and appropriate management become considerations. Until now, no information has been available on seizure reduction and human pregnancy related outcomes in women treated with KD therapies. MethodWe describe two cases of pregnant women with epilepsy treated with KD therapy either as monotherapy (Case 1) or as adjunctive therapy (Case 2). ResultsCase 1: A 27 year old woman, gravida1, started the classic KD with medium chain triglyceride (MCT) emulsion and 75g carbohydrate-restriction, later reduced to 47g. Glucose levels were 4–6mmol/L and blood ketone levels ranged from 0.2 to 1.4mmol/L. Seizure frequency decreased and seizure-free days increased. Mild side effects included intolerance to MCT, reduced serum carnitine and vitamin levels, and mild hyperlipidemia. Fetal and neonatal growth was normal as was growth and development at 12 months.Case 2: A 36 year-old nulliparous woman was treated with a 20 gram carbohydrate-restricted Modified Atkins Diet (MAD) and lamotrigine, resulting in reduction of seizure frequency to once per month prior to pregnancy. Once pregnant, carbohydrates were increased to 30g. When seizures increased, lamotrigine dose was doubled. Urine ketones trended down during second trimester. A male was born with bilateral ear deformities of unknown significance. The child had a normal neurodevelopment at eight months. ConclusionNon-pharmacological epilepsy therapies like KD and MAD may be effective during human pregnancy. However, safety still has to be established. Further monitoring to identify potential long term side effects is warranted.

SiRNA-mediated in vivo gene knockdown by acid-degradable cationic nanohydrogel particles

Cationic nanohydrogel particles have become an attractive tool for systemic siRNA delivery, but improvement of their in vivo tolerance is desirable, especially to prevent potential long term side effects by tissue and cellular accumulation. Here, we designed novel ketal cross-linked cationic nanohydrogel particles that were assessed for reduced tissue accumulation and robust siRNA delivery in vitro and in vivo. An oligo-amine cross-linker equipped with a ketal moiety in its core was synthesized and applied to nanohydrogel cross-linking of self-assembled reactive ester block copolymers in DMSO. The resulting acid-sensitive cationic nanoparticles spontaneously disassembled over time in acidic milieu, as investigated by dynamic light scattering. Fluorescent correlation spectroscopy showed effective complexation with siRNA as well as its release upon particle degradation at endosomal pH. These properties resulted in an enhanced in vitro gene knockdown for the acid-degradable cationic nanoparticles compared to their non-degradable spermine analogues. In a murine liver fibrosis model enhanced carrier and payload accumulation in the fibrotic tissue facilitated sequence-specific gene knockdown and prevented fibrosis progression. Long-term monitoring of the carrier in the body showed an enhanced clearance for the acid-degradable carrier, even after multiple dosing. Therefore, these acid-degradable cationic nanohydrogel particles can be considered as promising siRNA carriers for in vivo purposes towards therapeutic applications.

Target Site Mutagenesis during Crispr/ Cas 9/Single-Stranded- Oligonucleotide Directed Gene Editing for Sickle Cell Anemia

IntroductionSickle Cell Disease (SCD) results from a simple substitution of valine for glutamic acid at codon 6 in the β globin gene, resulting in an AàT transition in the third position. The mutation results in the production of hemoglobin HbS which differs from the normal HbAin that it tends to polymerize into long strands that deform the erythrocyte. While a variety of traditional treatment regimens or reagents, such as hydroxyurea and chronic transfusions have been used widely, these therapies are wrought with short and long term side effects that limit efficacy. There is great interest in the hypothesis that the repair of a single nucleotide is facilitated by the combined action of CRISPR/Cas 9 and single-stranded oligonucleotides (ssODNs) could prove a significant therapeutic advance for sickle cell disease. CRISPR/Cas 9 induces a site-specific double-stranded break while the single-stranded oligonucleotide provides a DNA template to improve the rate of accurate genetic correction. There is a great effort to understand off-site mutagenesis caused by editing of DNA regions remote to the target sequence. It is critical that investigators understand the frequency and types of DNA alterations induced by the gene editing reaction and affecting the region surrounding the targeted nucleotide site (herein termed on-site mutagenesis).MethodsWe targeted beta globin genes with a CRISPR/Cas9 system designed to cleave 2 bases to the 5'side of the targeted (A) nucleotide (the PAM site is located 2 bases to the 3' side on the complimentary strand) coupled to the electroporation of a single-stranded oligonucleotide (72-mer) designed to convert the wild-type (A) to the mutant (T) nucleotide. To evaluate the rate of on-site mutagenesis among individual alleles, we clonally expanded populations of edited K562 cells. We hypothesize that the evaluation of individual clones will permit a clear identification of intended and un-intended on-site DNA alteration. Allelic heterogeneity is analyzed using Tracking of Indels by DEcompositon (TIDE) methodology combined with Sanger sequencing.ResultsWe isolated 26 clonal lines for continued expansion after evidence of CRISPR/Cas 9 plasmid uptake and activity. Sanger sequencing with TIDE analysis revealed that the DNA sequence surrounding the targeted base is altered significantly as a result of the CRISPR/Cas9 gene editing process. Twenty three percent of the clones contain at least one corrected allele but one hundred percent of the clones exhibited mutagenicity of the DNA sequence surrounding the targeted base. All clones analyzed displayed varying degrees of sequence alteration (Figure 1). Interestingly, one clone contained a DNA insertion homologous to a region of the delta globin gene, suggesting that gene editing of the beta globin gene may have been repaired, in part, by delta globin DNA. The sequence of the delta globin locus was not changed.ConclusionsTaken together, our data suggest that combinatorial approaches to beta globin gene editing using CRISPR/Cas 9 and single-stranded oligonucleotides induce significant onsite mutagenesis and potential genetic swapping between related members of the same gene family. These observations provide insight into the type of molecular activity that accompanies combinatorial gene editing, particularly surrounding the target site.This work is supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number P20GM109021 [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Over-using chemotherapy in the adjuvant setting

Avoidance of unnecessary or ineffective treatment should be one of the main goals in adjuvant breast oncology today. Unfortunately, both patients and doctors hunt for tiny statistical differences in survival curves. This search could not only lead to an oncological approach of unlimited addition that we will not be able to afford, but would also end inevitably in indeterminate overtreatment with substantial risks of unexpected toxic effects eating away whatever progress we might make. “Do not harm” remains the main principle in medicine. To be able to follow this rule, we need to better understand the biology of breast cancer. The mistake of “one treatment fits all” can only be changed when we critically review trial designs of adjuvant breast oncology. The risk of overtreatment is there and selection of precisely defined cohorts for phase 3 trials is necessary, despite pressure of scientific ambition, pragmatism, and demands of industry. The “add on” clinical trial design model accepts the inability to confirm that standard therapy is still necessary if a positive result from the addition of the new therapy is obtained. The same model can be applied to “extended” adjuvant treatments in breast cancer subtypes. Addition of “miraculin” to the standard of care should generate a new standard. Such trials that show a modest benefit on average at a population level take us a step away from refining care for the individual, and might support the use of multiple and costly interventions with potential short and long term side effects. It is essential to escalate treatment when necessary and to de-escalate when un-necessary.

Syncope Following Sofosbuvir Therapy for Chronic Hepatitis C: A Report of Two Cases

The recently approved direct acting antiviral drug, Sofosbuvir‎, represents a revolution in treatment of chronic hepatitis C infection. Little is known about its long term side effects. Recent reports had drawn the attention to the possible cardio-toxic effect of sofosbuvir when combined with amiodarone. In our cases we reported occurrence of syncopal attacks in two cases few days after administration of sofosbuvir. Syncope was triggered by suddenly standing up from setting position in one patient and by micturition in the other. We suggest the occurrence of autonomic disturbance following sofosbuvir intake, an observation which need further post marketing evaluation in a large number of cases to clarify the cardio-autonomic effects of sofosbuvir.

The Association Between Testosteron Levels and Relationship Quality in Reproductive Aged Couples

<p>Couple relationship is often interfered by disorder in sexual activities. Most of the problems are in women than men. One of the main factor tha contributed to sexual function are sex hormones. Sex hormones selectively responsive to sexual incentives inducing a neurochemical state that favourable to sexual response. Androgens play an important role in sexual desire, arousal, orgasm and satisfaction by interacting with receptors in the hypothalamus, together with the dopaminergic, serotoninergic and opiatergic path, and the receptor genitals. Testosterone, molecular weight of 288.41 Dalton, is one of sex steroid hormones. It is the main androgenic hormone produced by the interstitial cells (Leydig). However, testosterone is an important precursor for the production of estradiol in the target tissue. Both testosteron and estrogen may affect sexual arousal. Decrease of testoterone in men is also related to declines in sexual desire, which can be restored with testosteron administration. Therefore, adding testosteron to estrogen in sexual disorder may be benefited. However, this practice was not widely use. Furhter sutdy is needed to assess its long term side effect.</p>

Anticancer chemotherapy in teenagers and young adults: managing long term side effects.

#### What you need to know Cancer is the leading cause of disease related death in teenagers and young adults (TYAs) in Western countries.1 2 In the UK, cancer in TYAs accounts for 9% and 15% of all male and female deaths respectively, and its incidence has risen by 19% since the mid-1990s, leading to 2300 new cases a year between 2011 and 2013.1 In Japan it accounted for nearly 7000 deaths between 2000 and 2006.3 TYA cancer survivors are likely to live for many decades but are at risk of late effects of their treatment. This article provides information for generalists on late effects of anticancer chemotherapy (see infographic and supplementary table A) that may affect quality of life. Radiotherapy related effects are not discussed but are summarised elsewhere.4 There is no internationally accepted age definition for TYAs. In the UK, TYA age is 15-24 years, whereas the US National Cancer Institute defines adolescents and young adults as aged 15-39 years. In this review we use the UK definition of 15-24 years. #### Sources and selection criteria This article is an evidence based review of late effects of chemotherapy in teenagers and young adults (TYAs). Our review encompasses studies using all age ranges termed as TYA or adolescents and young adult (AYA). We searched PubMed and the Cochrane databases between 1990 and 2016 using the search terms “teenage and young …

Mechanical and neuromuscular changes with lateral trunk lean gait modifications

Lateral trunk lean (LTL) is a proposed intervention for knee osteoarthritis but increased muscular demands have not been considered. The objective was to compare lower extremity and trunk muscle activation and joint mechanics between normal and increased LTL gait in healthy adults. Participants (n=20, mean age 22 years) were examined under two gait conditions: normal and increased LTL. A motion capture system and force plates sampled at 100 and 2000Hz respectively were used to determine joint angles and external moments including LTL angle and external knee adduction moment (KAM). Surface electromyography, sampled at 2000Hz, measured activation of six trunk/hip muscles bilaterally. Peak LTL angle, peak KAM, gait speed, and mean values from electromyography waveforms were compared between normal and LTL conditions using paired t-tests or 2-way analysis of variance. There was a significant (p<0.05) increase in peak LTL angle, decrease in first but not second peak KAM, and decrease in gait speed during LTL gait. There were significant (p<0.01) increases in external oblique and iliocostalis muscle activation during LTL gait. There was no change in activation for internal oblique, rectus abdominis, longissimus, and gluteus medius. LTL gait decreased early/mid-stance KAM demonstrating its ability to decrease medial compartment knee loading. Increases in external oblique and iliocostalis activation were present but small to moderate in size and unlikely to lead to short term injury. Longitudinal studies should evaluate the effectiveness of increased LTL for knee osteoarthritis and if the increase in muscular demands leads to negative long term side effects.

Abstract 1340: Dextran-Catechin conjugate: An anticancer nano-modified natural compound targeting copper metabolism in neuroblastoma

Abstract Background: Neuroblastoma is an aggressive childhood cancer that is poorly responsive to therapy. Moreover, survivors experience long term side effects from their treatment highlighting the need for effective and less toxic therapies. Catechin is a natural polyphenol with anti-cancer properties and limited side effects. Low serum stability of Catechin limits its clinical use that we overcame by chemical functionalization with Dextran. However its mechanism of action is unknown. Here, we investigated the mechanism of action of Dextran-Catechin, and its efficacy against neuroblastoma in vitro and in vivo. Methods: Anticancer activity was tested in 4 independent neuroblastoma cell lines using the cell viability assay Alamar Blue and apoptosis was assessed using PARP cleavage. Gene expression was determined using qRT-PCR and protein expression of CTR1 by western blotting. Intracellular copper was measured by spectrophotometric analysis. Fluorescence-lifetime imaging microscopy was used to study NADH/NAD+ ratio to determine the induction of oxidative stress. Cellular levels of the antioxidant GSH was examined using a colorimetric assay. PET imaging studies with Cu64 were performed in a xenograft neuroblastoma model to monitor copper uptake in tumors. In vivo anticancer activity of Dextran-Catechin was assessed in human xenograft and syngeneic models of neuroblastoma. Results: The neuroblastoma cell lines SH-SY5Y, IMR-32, BE(2)C and doxorubicin-resistant BE(2)C-ADR were sensitive to Dextran-Catechin (IC50 9.7 μg/ml, 17.83 μg/ml, 16 μg/ml and 18.2 μg/ml, respectively) at concentrations that were not toxic to non-malignant MRC-5 cells. However, Cisplatin-resistant neuroblastoma cells, IMR-32-CisRes, were 2.5-fold resistant against Dextran-Catechin compared to the parental cells. Copper transporter 1 (CTR1) mediates cisplatin uptake and IMR-32-CisRes exhibited 50% lower expression of CTR1 and lower intracellular copper compared to the IMR-32 cells. In contrast, Dextran-Catechin sensitive neuroblastoma cells had elevated intracellular copper implicating copper in the activity of this conjugate. We demonstrated that Dextran-Catechin reacts with copper generating reactive oxygen species and inducing cancer cell death. Dextran-Catechin treatment caused a decrease of NADH/NAD+ ratio and GSH levels, confirming oxidative stress. PET imaging analysis of the IMR-32-neuroblastoma xenograft model revealed high accumulation of copper in the tumor mass. The high levels of copper were maintained in the tumor mass for up to 48h, while Cu64 was cleared by the other organs. Importantly, we showed that Dextran-Catechin significantly reduced tumor growth in human xenograft and syngeneic models of neuroblastoma with no evidence of side effects. Conclusion: Dextran-Catechin targets copper, inhibits tumour growth, and has therapeutic potential for cancers dependent on copper for their growth. Citation Format: Orazio Vittorio, Miriam Brandl, Giuseppe Cirillo, Kathleen Kimpton, Elizabeth Hinde, Hien T. T. Duong, Cyrille Boyer, Claudia Flemming, Eugene M.H. Yee, Naresh Kumar, Arvind Parmar, Giancarlo Pascali, Arnaud Charil, Michelle Haber, Murray Norris, Maria Kavallaris. Dextran-Catechin conjugate: An anticancer nano-modified natural compound targeting copper metabolism in neuroblastoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1340.