bluuebunny/crossref_metadata_2025 huggingface dataset:
Reassessing the Role of the Active TGF-β1 as a Biomarker in Systemic Sclerosis: Association of Serum Levels with Clinical Manifestations
Authors: Dantas Andréa Tavares , Gonçalves Sayonara Maria Calado , Almeida Anderson Rodrigues de , Gonçalves Rafaela Silva Guimarães , Sampaio Maria Clara Pinheiro Duarte , Vilar Kamila de Melo , Pereira Michelly Cristiny , Rêgo Moacyr Jesus Barreto de Melo , Pitta Ivan da Rocha , Marques Claudia Diniz Lopes , Duarte Angela Luzia Branco Pinto , Pitta Maira Galdino da Rocha
Published: November 2016

Objective. To determine active TGF-β1 (aTGF-β1) levels in serum, skin, and peripheral blood mononuclear cell (PBMC) culture supernatants and to understand their associations with clinical parameters in systemic sclerosis (SSc) patients.Methods. We evaluated serum samples from 56 SSc patients and 24 healthy controls (HC). In 20 SSc patients, we quantified spontaneous or anti-CD3/CD28 stimulated production of aTGF-β1 by PBMC. The aTGF-β1 levels were measured by ELISA. Skin biopsies were obtained from 13 SSc patients and six HC, and TGFB1 expression was analyzed by RT-PCR.Results. TGF-β1 serum levels were significantly higher in SSc patients than in HC (p< 0.0001). Patients with increased TGF-β1 serum levels were more likely to have diffuse subset (p= 0.02), digital ulcers (p= 0.02), lung fibrosis (p< 0.0001), positive antitopoisomerase I (p= 0.03), and higher modified Rodnan score (p= 0.046). Most of our culture supernatant samples had undetectable levels of TGF-β1. No significant difference in TGFB1 expression was observed in the SSc skin compared with HC skin.Conclusion. Raised active TGF-β1 serum levels and their association with clinical manifestations in scleroderma patients suggest that this cytokine could be a marker of fibrotic and vascular involvement in SSc.
Lymphocyte Antigen 75 Polymorphisms Are Associated with Disease Susceptibility and Phenotype in Japanese Patients with Inflammatory Bowel Disease
Authors: Hirayama Atsuhiro , Joshita Satoru , Kitahara Kei , Mukawa Kenji , Suga Tomoaki , Umemura Takeji , Tanaka Eiji , Ota Masao
Published: November 2016

Recent genome-wide association studies have rapidly improved our understanding of the molecular pathways leading to inflammatory bowel disease (IBD), which includes Crohn’s disease (CD) and ulcerative colitis (UC). Although several reports have demonstrated that gene single nucleotide polymorphisms (SNPs) are associated with susceptibility to IBD, its precise genetic factors have not been fully clarified. Here, we performed an association analysis between lymphocyte antigen 75 (LY75) genetic variations and IBD susceptibility or phenotype. SNPs were genotyped in 51 CD patients, 94 UC patients, and 269 healthy controls of Japanese ethnicity. We detected a significant relationship with CD susceptibility for the rs16822581 LY75 SNP (P=0.045). One haplotype (GT, P=0.042) was also associated with CD susceptibility, while another carrying the opposite SNP (CA) was linked to an absence of surgical history for CD. Our findings confirm that LY75 is involved in CD susceptibility and may play a role in disease activity in the Japanese population.
Rebamipide, an Amino Acid Analog of 2(1H)-Quinolinone, Inhibits the Formation of Human Osteoclasts
Authors: Nanke Yuki , Kobashigawa Tsuyoshi , Yago Toru , Kawamoto Manabu , Yamanaka Hisashi , Kotake Shigeru
Published: November 2016

Objectives. Drug repositioning or drug reprofiling (DR) has recently been growing in importance. DR has a significant advantage over traditional drug development because the repositioned drug has already passed toxicity tests; its safety is known, and the risk of adverse toxicology is reduced. In the current study, we investigated the role of rebamipide, a mucosa-protecting agent, with recently reported anti-inflammatory function, in human osteoclastogenesis. Methods. Peripheral blood mononuclear cells (PBMCs) were cultured in the presence of M-CSF and sRANKL. Osteoclast formation was evaluated by immunohistological staining for CD51/61 (vitronectin receptors). Osteoclast formation, in the presence or absence of rebamipide (0, 1, and 3 mM), was observed by time-lapse photography and actin ring formation. The number of absorption sites and area of absorption were calculated using Osteologic™ plates. Pit formation was studied by 3D-SEM. Results. Rebamipide inhibited human osteoclast formation at 3 mM, a pharmacological concentration, and inhibited resorbing activity dose-dependently. Rebamipide induced the degradation of actin rings in mature osteoclasts. This mechanism may involve inhibiting the osteoclast fusion pathway through reducing the expression of DC-specific transmembrane protein (DC-STAMP). Conclusions. The present study suggests that rebamipide would be useful as a novel agent for osteoporosis and rheumatoid arthritis.
Risk Factors and Symptoms of Meibomian Gland Loss in a Healthy Population
Authors: Machalińska Anna , Zakrzewska Aleksandra , Safranow Krzysztof , Wiszniewska Barbara , Machaliński Bogusław
Published: November 2016

Purpose. The aim of this study was to investigate the relationships between MGL and ocular symptoms, several systemic conditions, and key markers of ocular surface health.Methods. We included into the study 91 healthy volunteers between the ages of 20 and 77 years. We analyzed meibomian gland morphology, function, and lid margin alterations. We correlated our findings with self-reported ocular symptoms, systemic medical history, lifestyle factors, and tear film abnormalities.Results. We observed that a high ocular surface disease index, a history of either chalazion or hordeolum, experience of puffy eyelids upon waking, and foreign body sensation all appeared to be predictors of an abnormal meiboscore after adjusting for age and sex (p=0.0007;p=0.001;p=0.02;p=0.001, resp.). Multivariate logistic regression model including age and sex showed that there were three independent predictors of abnormal meiboscore: older age (OR = 1.03, 95% CI = 1.01–1.04 per year,p=0.006), postmenopausal hormone therapy (OR = 4.98, 95% CI = 1.52–16.30,p=0.007), and the use of antiallergy drugs (OR = 5.85, 95% CI = 2.18–15.72,p=0.0004).Conclusion. Our findings extend current knowledge on the pathophysiology of MGL.
Sinus Venosus Atrial Septal Defect Complicated by Eisenmenger Syndrome and the Role of Vasodilator Therapy
Authors: Anuwatworn Amornpol , Gedela Maheedhar , Bendaly Edgard , Prescott-Focht Julia A. , Yee Jimmy , Clark Richard , Jonsson Orvar
Published: November 2016

Sinus venosus atrial septal defect is a rare congenital, interatrial communication defect at the junction of the right atrium and the vena cava. It accounts for 5–10% of cases of all atrial septal defects. Due to the rare prevalence and anatomical complexity, diagnosing sinus venous atrial septal defects poses clinical challenges which may delay diagnosis and treatment. Advanced cardiac imaging studies are useful tools to diagnose this clinical entity and to delineate the anatomy and any associated communications. Surgical correction of the anomaly is the primary treatment. We discuss a 43-year-old Hispanic female patient who presented with dyspnea and hypoxia following a laparoscopic myomectomy. She had been diagnosed with peripartum cardiomyopathy nine years ago at another hospital. Transesophageal echocardiography and computed tomographic angiography of the chest confirmed a diagnosis of sinus venosus atrial septal defect. She was also found to have pulmonary arterial hypertension and Eisenmenger syndrome. During a hemodynamic study, she responded to vasodilator and she was treated with Ambrisentan and Tadalafil. After six months, her symptoms improved and her pulmonary arterial hypertension decreased. We also observed progressive reversal of the right-to-left shunt. This case illustrates the potential benefit of vasodilator therapy in reversing Eisenmenger physiology, which may lead to surgical repair of the atrial septal defect as the primary treatment.
Silkworm Sericin: Properties and Biomedical Applications
Authors: Kunz Regina Inês , Brancalhão Rose Meire Costa , Ribeiro Lucinéia de Fátima Chasko , Natali Maria Raquel Marçal
Published: November 2016

Silk sericin is a natural polymer produced by silkworm,Bombyx mori, which surrounds and keeps together two fibroin filaments in silk thread used in the cocoon. The recovery and reuse of sericin usually discarded by the textile industry not only minimizes environmental issues but also has a high scientific and commercial value. The physicochemical properties of the molecule are responsible for numerous applications in biomedicine and are influenced by the extraction method and silkworm lineage, which can lead to variations in molecular weight and amino acid concentration of sericin. The presence of highly hydrophobic amino acids and its antioxidant potential make it possible for sericin to be applied in the food and cosmetic industry. The moisturizing power allows indications as a therapeutic agent for wound healing, stimulating cell proliferation, protection against ultraviolet radiation, and formulating creams and shampoos. The antioxidant activity associated with low digestibility of sericin that expands the application in the medical field, such as antitumour, antimicrobial and anti-inflammatory agent, anticoagulant, acts in colon health, improving constipation and protects the body from obesity through improved plasma lipid profile. In addition, the properties of sericin allow its application as a culture medium and cryopreservation, in tissue engineering and for drug delivery, demonstrating its effective use, as an important biomaterial.
The Role of the Cognitive Control System in Recovery from Bilingual Aphasia: A Multiple Single-Case fMRI Study
Authors: Radman Narges , Mouthon Michael , Di Pietro Marie , Gaytanidis Chrisovalandou , Leemann Beatrice , Abutalebi Jubin , Annoni Jean-Marie
Published: November 2016

Aphasia in bilingual patients is a therapeutic challenge since both languages can be impacted by the same lesion. Language control has been suggested to play an important role in the recovery of first (L1) and second (L2) language in bilingual aphasia following stroke. To test this hypothesis, we collected behavioral measures of language production (general aphasia evaluation and picture naming) in each language and language control (linguistic and nonlinguistic switching tasks), as well as fMRI during a naming task at one and four months following stroke in five bilingual patients suffering from poststroke aphasia. We further applied dynamic causal modelling (DCM) analyses to the connections between language and control brain areas. Three patients showed parallel recovery in language production, one patient improved in L1, and one improved in L2 only. Language-control functions improved in two patients. Consistent with the dynamic view of language recovery, DCM analyses showed a higher connectedness between language and control areas in the language with the better recovery. Moreover, similar degrees of connectedness between language and control areas were found in the patients who recovered in both languages. Our data suggest that engagement of the interconnected language-control network is crucial in the recovery of languages.
A Previously Undescribed Presentation of Mixed Adenoneuroendocrine Carcinoma
Authors: De Luca-Johnson Javier , Zenali Maryam
Published: November 2016

We report a case of mixed adenoneuroendocrine carcinoma (MANEC) of stomach with tubular adenoma and well-differentiated neuroendocrine tumor (WD-NET) in the primary tumor in the stomach giving rise to biphenotypic regional nodal metastases. A 35-year-old woman with abdominal pain was found to have a 1.8-cm gastric lesion, diagnosed as WD-NET (intermediate grade) on the biopsy. The resection specimen contained residual WD-NET; there was also a gastric adenoma adjacent to the NET and nodal metastasis with both adeno- and neuroendocrine components. The tumor was classified as MANEC. Of note, the entire gastric tissue was submitted and multiple deeper levels of the adenomatous lesion were examined; no adenocarcinoma was present in the primary lesion. While association of gastric adenoma with neuroendocrine neoplasm is rare, presence of biphenotypic metastasis originating from such a lesion is highly unusual and to the best of our knowledge has not been reported.
Magnitude of Maternal Anaemia in Rural Burkina Faso: Contribution of Nutritional Factors and Infectious Diseases
Authors: Meda Nicolas , Coulibaly Malik , Nebie Yacouba , Diallo Ibrahima , Traore Yves , Ouedraogo Laurent T.
Published: November 2016

Background. Maternal anaemia is a worldwide public health problem affecting particularly developing countries. In Burkina Faso, little data is available for rural areas. This study aimed to determine the prevalence of maternal anaemia and the risk factors associated with it in the rural health district of Hounde in Burkina Faso but also to define better control measures of maternal anaemia.Methods. This cross-sectional study conducted in 2010 had a sample of 3,140 pregnant women attending antenatal care in all the 18 primary health care facilities of the district. The women’s characteristics and their knowledge about contraceptives and sexually transmitted infections (STI) were collected. Also, physical and gynaecological examination, completed by vaginal, cervix, blood, and stool samplings, were collected.Results. A prevalence of 63.1% was recorded for maternal anaemia. Geophagy rate was 16.3% and vitamin A deficiency 69.3%. In addition, anaemia was independently associated with low education, low brachial perimeter, geophagy, and primigravida. But no statically significant relationship was found between maternal anaemia and infectious diseases or vitamin A deficiency.Conclusion. The magnitude of maternal anaemia was found to be higher in rural Hounde health district and should be addressed by adequate policy including education and the fight against malnutrition.
n-Gram-Based Text Compression
Authors: Nguyen Vu H. , Nguyen Hien T. , Duong Hieu N. , Snasel Vaclav
Published: November 2016

We propose an efficient method for compressing Vietnamese text usingn-gram dictionaries. It has a significant compression ratio in comparison with those of state-of-the-art methods on the same dataset. Given a text, first, the proposed method splits it inton-grams and then encodes them based onn-gram dictionaries. In the encoding phase, we use a sliding window with a size that ranges from bigram to five grams to obtain the best encoding stream. Eachn-gram is encoded by two to four bytes accordingly based on its correspondingn-gram dictionary. We collected 2.5 GB text corpus from some Vietnamese news agencies to buildn-gram dictionaries from unigram to five grams and achieve dictionaries with a size of 12 GB in total. In order to evaluate our method, we collected a testing set of 10 different text files with different sizes. The experimental results indicate that our method achieves compression ratio around 90% and outperforms state-of-the-art methods.
Effect of Hydrogen Peroxide Content on the Preparation of Peroxotitanate Materials for the Treatment of Radioactive Wastewater
Authors: Yang Wein-Duo , Nam Chau Thanh , Chung Jen-Chien , Huang Hsin-Ya
Published: November 2016

The modification of peroxotitanate using hydrogen peroxide significantly improved the ion-exchange capacity of titanate materials as sorbents for metal ions contained in a radioactive waste simulant solution. The effects of hydrogen peroxide content (hydrogen peroxide/titanium isopropoxide molar ratios, hereafter expressed as H/T) on the properties of as-prepared titanate synthesized at 130°C and at pH of 6-7, followed by freeze-drying, were investigated. The peroxotitanate materials thus obtained were characterized by XRD, BET, SEM, TEM, EDX, ICP, and Raman spectroscopy. At an H/T ratio of 2, peroxotitanate predominantly exhibited an amorphous structure, with a clearly observed tubular or fibrous structure. Furthermore, peroxotitanate modified at an H/T ratio of 2 exhibited the best ion-exchange capacity of 191 mg g−1 for metal ions contained in a radioactive waste simulant solution. Hence, these peroxotitanate materials are suitable for removing metal ions from wastewater, especially lanthanide ions (Ln3+) and Sr2+.
hsa-miR-376c-3p Regulates Gastric Tumor Growth BothIn VitroandIn Vivo
Authors: Tu Lin , Zhao Enhao , Zhao Wenyi , Zhang Zizhen , Tang Defeng , Zhang Yeqian , Wang Chaojie , Zhuang Chun , Cao Hui
Published: November 2016

Background. In recent studies, aberrant expression of various microRNAs (miRNAs) is reported to be associated with gastric cancer metastasis.Method. Overexpression construct and inhibitor of hsa-miR-376c-3p were expressed in human gastric adenocarcinoma cell line SGC-7901. The expression level of tumor related genes was detected by qPCR, western blot, and immunostaining. Cell apoptosis was determined by flow cytometry. Xenograft of SGC-7901 cells was used to elucidate the function of hsa-miR-376c-3p in gastric tumor growthin vivo.Result. Expression of hsa-miR-376c-3p was detected in SGC-7901 cells. Downregulation of hsa-miR-376c-3p increased the expression level of BCL-2 and decreased the expression of smad4 and BAD. On the contrary, overexpression of hsa-miR-376c-3p increased the expression of BAD and smad4, while it led to the decreasing expression level of BCL-2. Overexpression of hsa-miR-376c-3p also promoted cell apoptosisin vitroand inhibited gastric tumor growthin vivo. Furthermore, the expression of BCL-2 was higher and expression of smad4 and BAD was lower in tumor tissue than the tissue adjacent to tumor from gastric cancer patients.Conclusion. This study demonstrated that hsa-miR-376c-3p plays an important role in the inhibition of gastric tumor growth and tumor related gene expression bothin vitroandin vivo.
Abstract B04: Novel Methods to Target RB Pathway Disruption in Osteosarcoma
Authors: Hinds Philip W. , Enos Miriam , Kong Elizabeth , Dignan Elisabeth , Gunduz Volkan , Pietruska Jodie
Published: November 2016

Osteosarcoma, prevalently a pediatric disease, is the most common primary non-hematologic bone tumor. The five-year survival rates range from 24-62% depending on age. A deeper understanding of the disease and new therapeutic approaches are crucial to reducing osteosarcoma's morbidity and mortality.

Data from our and other laboratories point to interconnections between bone development and cancer. Specifically, RB1 gene mutation (encoding retinoblastoma protein or pRB) is observed in 60% of sporadic osteosarcomas, and the pRB pathway is altered in osteosarcomas that retain pRB function. Mouse data suggests that loss of pRB can change osteoprogenitor function and increase osteosarcoma formation. Our work shows that: pRB loss leads to the expansion of bipotent (adipogenic/osteogenic) progenitor cells, properties also found in osteosarcomas; pRB binds to and activates the transcriptional regulator of osteogenesis Runx2, suggesting that pRB could alter promoter binding by chromatin modifying enzymes and regulate osteoblast differentiation; pRB can act as a transcriptional coactivator with Runx2 inducing Indian hedgehog (Ihh) expression late in osteoblast differentiation (Ihh can drive osteoprogenitor cells to the osteoblast lineage, potentially mediating the pRB phenotype loss in osteoblasts). Our studies also show a role for CDK6 in Runx2 activation and bone development, and a requirement for CDK6 activity in p53-dependent osteosarcoma. Thus, here we (a) analyze the role of pRB in fate specification of osteoblasts; (b) test the function of Ihh as a mediator of altered differentiation and tumorigenesis in pRB-null cells; and (c) test the role of CDK6 as a therapeutic target in osteosarcoma.

Citation Format: Philip W. Hinds, Miriam Enos, Elizabeth Kong, Elisabeth Dignan, Volkan Gunduz, Jodie Pietruska. Novel Methods to Target RB Pathway Disruption in Osteosarcoma. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr B04.
Abstract B06: Single-cell RNA-seq profiling of transcriptional transition states during human retinoblastoma development
Authors: Lee Sunhye , Singh Hardeep , Cobrinik David
Published: November 2016

Retinoblastoma is a rare childhood tumor initiated by biallelic inactivation of the RB1 gene and loss of function of retinoblastoma (Rb) protein. Although Rb loss is a key-initiating event, the molecular mechanisms controlling the transformation into malignancy remain unclear. In this project, we seek to define the cell state transitions that follow RB1 inactivation in a defined cell of origin, the cone photoreceptor precursor. Previously we revealed that Rb knockdown could induce cone precursors to proliferate in vitro and to form retinoblastoma-like tumors in orthotopic xenografts. Using post-mitotic cone precursors from normal human fetal retina, we describe the generation, isolation, and single-cell RNA sequencing (RNA-seq) on RB1-knockdown cells at multiple time points to profile the levels of all polyadenylated transcripts. We then use “Monocle” to pseudo-temporally order cells according to their transcriptome profiles into distinct states through which the cells pass during cell cycle entry and tumor progression. Through this single cell RNA-seq approach, we will define the transcriptional alteration events of each cell state transition, and identify potential mediators of such transitions during retinoblastoma development. In addition, we define effects of inhibitors targeting potential mediators of each cell state transition as therapeutic candidates, currently focusing on Nutlin-3a (targeting MDM2), JQ1 (targeting MYCN), and SNS-032 and SU9516 (targeting CDK2). Taken together, this study seeks to identify transformation-related events that can be pharmacologically targeted to suppress tumorigenesis in genetically predisposed children.

Citation Format: Sunhye Lee, Hardeep Singh, David Cobrinik. Single-cell RNA-seq profiling of transcriptional transition states during human retinoblastoma development. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr B06.
Abstract B15: Synergistic functions of E2F7 and E2F8 are critical to suppress stress induced skin cancer
Authors: Westendorp Bart , Thurlings Ingrid , Martinez-Lopez Lina , Zijp Marjolein , Kuiper Raoul , Tooten Peter , Vos HarmJan , Burgering Boudewijn , Bruin Alain de
Published: November 2016

E2F transcription factors are important regulators of the cell cycle, and unrestrained activation of E2F-dependent transcription is considered to be an important driver of tumor formation and progression. Although highly expressed in normal skin and skin cancer, the role of the atypical E2Fs, E2F7 and E2F8, in keratinocyte homeostasis, regeneration and tumorigenesis is unknown. Surprisingly, keratinocyte-specific deletion of E2F7 and E2F8 in mice did not interfere with skin development and wound healing. However, the rate for successful isolation and establishment of E2f7/8-deficient primary keratinocyte cultures was much higher than for wild-type keratinocytes. Moreover, E2f7/8-deficient primary keratinocytes proliferate more efficiently under stress conditions, such as low/high confluence or DNA damage. Application of in vivo stress using the DMBA/TPA skin carcinogenesis protocol revealed that combined inactivation of E2f7/8 enhanced tumorigenesis and accelerated malignant progression. Loss of atypical E2Fs resulted in increased expression of E2F target genes, including E2f1. Additional loss of E2f1 did not rescue, but worsened skin tumorigenesis. We show that loss of E2F7/8 triggers apoptosis via induction of E2F1 in response to stress, indicating that the tumor-promoting effect of E2F7/8 inactivation can be partially compensated via E2F1-dependent apoptosis. Importantly, E2F7/8 repressed a large set of E2F target gene that are highly expressed in human patients with skin cancer. Together, our studies demonstrate that atypical E2Fs act as tumor suppressors, most likely via transcriptional repression of cell cycle genes in response to stress.

Citation Format: Bart Westendorp, Ingrid Thurlings, Lina Martinez-Lopez, Marjolein Zijp, Raoul Kuiper, Peter Tooten, HarmJan Vos, Boudewijn Burgering, Alain de Bruin. Synergistic functions of E2F7 and E2F8 are critical to suppress stress induced skin cancer. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr B15.
Abstract B30: Resistance to Photodynamic Therapy in Non-Melanoma Skin Cancer Cells
Authors: Leon Daniela , Silva Ramon , Inada Natalia , Kurachi Cristina , Ili Carmen Gloria , Brebi Priscilla , Roa Juan Carlos
Published: November 2016

Introduction: Squamous skin cancer is one of the most common neoplasia worldwide and may be preceded by Bowen's disease (BD) and Actinic Keratosis (AK). These preneoplastic lesions can be treated with photodynamic therapy (PDT). PDT causes cell death mainly by reactive oxygen species (ROS) produced through a combination of O2, a photosensitizer and light. BD and AK have well overcomes after PDT. However, high recurrence rates of some lesions have been reported.

Objective: To generate resistant cells to PDT as an in vitro model for future studies.

Methodology: HSC-1 cells from human squamous cell cancer (SCC) of skin were used. Cells were treated with a total of 16 cycles of PDT. Every PDT cycle was carried out incubating cells with methyl aminolevulinate (MAL), as a protoporphyrin IX (PpIX) precursor, for 4 hours in the dark. Then, cells were irradiated with red light (630 nm). The light fluence used for the four first cycles was 1 J/cm2, for subsequent cycles light fluence increased to 2, 3 and 4 J/cm2. Cell viability was evaluated 1 and 24 hours after PDT by flow cytometry and MTT assay respectively. Intracellular PpIX was detected by flow cytometry and the PpIX content was quantified using a standard curve. By flow cytometry using a CM-H2DCFDA probe intracellular ROS was detected.

Results: After 24 hours of PDT (4 J/cm2), viability in parental HSC-1 cells was 5%, meanwhile in resistant cells was 70% (P < 0.05). On the other hand, 88% of parental cells produced PpIX (40 ng/mg protein) and 18% of resistant cells produced PpIX (17 ng/mg protein) (P < 0.05). No significant differences were found in ROS detection in both parental and resistant cells.

Conclusion: The results indicate that it was possible to obtain HSC-1 cells two-fold resistant to PDT compared to their parental cells.

Acknowledgments: Financial support by CONICYT. CONICYT-PCHA/Doctorado Nacional/2012-21120665, CORFO 09CN14-5960, Corfo N°12IDL2-18157, FONDECYT N° 11150802, FONDECYT N°11150622.

Citation Format: Daniela Leon, Ramon Silva, Natalia Inada, Cristina Kurachi, Carmen Gloria Ili, Priscilla Brebi, Juan Carlos Roa. Resistance to Photodynamic Therapy in Non-Melanoma Skin Cancer Cells. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr B30.
Abstract IA02: Neomorphic functions of cyclin D1 during neoplastic development
Authors: Diehl J. Alan
Published: November 2016

Cyclin D1 is a cell cycle recipient of diverse oncogenic signals and a multifunctional transcriptional modulator. Extensive studies have characterized mutational disruption of p53 signaling in human cancers. However, the mechanisms underlying retention of wild-type p53 during neoplastic outgrowth have remained ambiguous. We have identified and functionally validated a crucial role for arginine-methylation of p53 as a mechanism for inhibition of p53-dependent apoptosis in response to oncogenic insults. Phosphorylation-dependent activation of PRMT5/MEP50 enforces catalytic targeting of p53 thereby reducing p53 occupancy on key target promoters and sustaining leukemia/lymphoma tumor cell proliferation and survival. The requisite role for PRMT5 in the context of multiple lymphoma/leukemia oncogenic drivers suggests a molecular rationale for therapeutic development.

Citation Format: J. Alan Diehl. Neomorphic functions of cyclin D1 during neoplastic development. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr IA02.
Abstract IA11: Transient CDK4/6 inhibition protects hematopoietic progenitors from chemotherapy-induced exhaustion
Authors: Sharpless Norman E.
Published: November 2016

Conventional cytotoxic chemotherapy is highly effective in certain cancers, but causes dose-limiting damage to normal proliferating cells, especially hematopoietic stem and progenitor cells (HSPCs). Serial exposure to cytotoxics causes a long-term hematopoietic compromise (“exhaustion”), which limits the use of chemotherapy and success of cancer therapy. I will discuss the use of small-molecule inhibitors of cyclin-dependent kinases 4 and 6 (CDK4/6) contemporaneously with DNA damaging stimuli such as cytotoxic chemotherapy to protect HSPC from chemotherapy-induced exhaustion. Specifically, CDK4/6 inhibitors will reduce HSPC proliferation, leading to decreased damage from cytotoxic agents, resulting in preserved HSPC function acutely after bone marrow injury as well as late time points well after hematopoietic recovery. In particular, I will show this approach leads to enhanced long-term HSC function resulting in enhanced serial transplantation and reduced myeloid skewing. I will discuss similarities between chemotherapy-induced HSC dysfunction and HSC aging. Moreover, these finding suggest that the combination of CDK4/6 inhibitors with DNA damaging chemotherapy provides a means to prevent therapy-induced bone marrow exhaustion.

Citation Format: Norman E. Sharpless. Transient CDK4/6 inhibition protects hematopoietic progenitors from chemotherapy-induced exhaustion. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr IA11.
Abstract PR04: Characterizing the sequence of cell-cycle events during proliferation and quiescence
Authors: Gookin Sara , Spencer Sabrina Leigh
Published: November 2016

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR04) of the Conference Proceedings.

Although the cell-cycle field has identified most of the key biochemical events involved in cell-cycle progression, we do not have a clear picture of overall cell-cycle dynamics. The prevalence of cell-to-cell variability is increasingly appreciated, but this heterogeneity is blurred when bulk analysis approaches are used, potentially resulting in incorrect interpretations of biological data. Using a live-cell sensor for CDK2 activity, we recently discovered that unperturbed cycling cells bifurcate into two subpopulations as they exit mitosis: one subpopulation begins to immediately increase CDK2 activity (CDK2inc cells), whereas the other subpopulation turns off CDK2 activity and enters a period of quiescence (CDK2low cells). These cells can later re-enter the cell cycle by building up CDK2 activity (CDK2low->inc). Here we use single-cell time-lapse microscopy followed immediately by immunofluorescence to determine the dynamics of over 15 cell-cycle proteins in unperturbed cycling mammary epithelial cells. By sorting individual cells by their CDK2inc versus CDK2low trajectory, and aligning them as a function of time-since-anaphase, we identified several unexpected differences in protein levels (or modification states) in the two subpopulations. Moreover, we identified certain proteins whose concentrations steadily decrease the longer the cells are in quiescence, suggesting the existence of multiple levels of quiescence depth. Cells can also emerge from quiescence by engaging CDK2 activity. When we align these CDK2low->inc cells to the time of initial increase in CDK2 activity, we recapitulate the cell-cycle dynamics seen in the CDK2inc cells. These results argue that cell-cycle commitment is not a point in time, but rather a molecular event (CDK2 activation) that varies temporally across cells. In summary, our single-cell measurements provide a critical resource for the field by describing the dynamics and sequence of many key cell-cycle proteins in unperturbed proliferating and quiescent cells.

This abstract is also being presented as Poster A05.

Citation Format: Sara Gookin, Sabrina Leigh Spencer. Characterizing the sequence of cell-cycle events during proliferation and quiescence. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr PR04.
Abstract PR07: Recruitment of Pontin/Reptin by E2F1 amplifies E2F transcriptional response during cancer progression
Authors: Tarangelo Amy , Lo Nathanael , Teng Rebecca , Kim Eunsun , Raman Pichai , Viatour Patrick
Published: November 2016

Unrestricted E2F activity and increased E2F1 expression are hallmarks of liver cancer, but the consequences of aberrant E2F activity for liver cancer progression remain ill defined. Our data show that aberrant E2f activity, following inactivation of the Rb gene family in a mouse model of liver cancer, initially triggers a robust gene expression program associated with the cell cycle and predominantly driven by E2f2 and E2f3. During liver cancer progression, slowly accumulating E2f1 recruits a Pontin/Reptin complex to insert the H2a.z histone variant and open the chromatin conformation at E2f1-bound target genes. This epigenetic mechanism amplifies the E2f transcriptional response, leading to enhanced transactivation of cell cycle genes and de novo activation of low binding affinity E2f target genes that regulate non-cell cycle oncogenic features, such as the Warburg effect. In addition, extensive mouse genetic approaches indicate that E2f-driven liver cancer initiation and progression is cell type specific, suggesting that a permissive chromatin environment is necessary to enable E2f oncogenic functions. Collectively, these data indicate that sustained E2F activity expands E2F transcriptional response and connects the regulation of cell cycle with multiple non cell cycle functions that drive tumor progression.

This abstract is also being presented as Poster B13.

Citation Format: Amy Tarangelo, Nathanael Lo, Rebecca Teng, Eunsun Kim, Pichai Raman, Patrick Viatour. Recruitment of Pontin/Reptin by E2F1 amplifies E2F transcriptional response during cancer progression. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr PR07.
Abstract PR11: APC/CCdh1 maintains primordial follicles, germinal vesicle arrest and ensures balanced segregation of chromosomes by enabling removal of Shugoshin-2 from chromosomes arms
Authors: Rattani Ahmed , Mejia Randy Ballestros , Roberts Katherine , Roig Maurici B , Godwin Jonathan , Eguren Manuel , Ogushi Sugako , Okaz Elwy , Pendas Alberto , Marcos Malumbres , Herbert Mary , Nasmyth Kim
Published: November 2016

Germ cells in mammals, including humans, remain arrested at the primordial follicle stage from a few days to many years depending on the species. At each hormonal cycle, a few of these cells grow in size to become fully-grown oocytes but remain arrested in meiotic prophase. A resumption of meiosis is heralded by germinal vesicle breakdown (GVBD), a gradual rise in Cdk1 activity, condensation of bivalent chromosomes, and their eventual alignment on metaphase plates with maternal and paternal kinetochore pairs pulled in opposite directions (co-orientation). The first meiotic division is triggered by activation of a form of the Anaphase-promoting complex/cyclosome associated with Cdc20 (APC/CCdc20), which activates separase and thereby destroys sister chromatid cohesion along chromosome arms, resolving chiasmata. Because cohesion around centromeres is protected from separase by protein phosphatase 2A (PP2A) recruited by shugoshin (Sgol2), bivalents are converted at this division to dyad chromosomes in which sister chromatids are held together by peri-centric cohesin. We show here that an alternative version of the APC/C associated with the Cdc20-like Cdh1 protein (APC/CCdh1) has equally important roles. By promoting proteolysis of mitotic cyclins and Cdc25B, APC/CCdh1 delays the increase in Cdk1 activity leading to GVBD. More surprising, by moderating the rate at which Cdk1 is activated following GVBD, APC/CCdh1 creates conditions necessary for the removal of Sgol2 from chromosome arms by the Aurora B/C kinase, an event crucial for the efficient resolution of chiasmata. Lastly, by targeting proteins other than cyclins, APC/CCdh1 prevents apoptosis of primordial follicles during their extended prophase (G2) arrest.

This abstract is also being presented as Poster A24.

Citation Format: Ahmed Rattani, Randy Ballestros Mejia, Katherine Roberts, Maurici B Roig, Jonathan Godwin, Manuel Eguren, Sugako Ogushi, Elwy Okaz, Alberto Pendas, Malumbres Marcos, Mary Herbert, Kim Nasmyth. APC/CCdh1 maintains primordial follicles, germinal vesicle arrest and ensures balanced segregation of chromosomes by enabling removal of Shugoshin-2 from chromosomes arms. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Cancer Cell Cycle - Tumor Progression and Therapeutic Response; Feb 28-Mar 2, 2016; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(11_Suppl):Abstract nr PR11.
Docosahexaenoic Acid
Authors: Calder Philip C.
Published: November 2016

Docosahexaenoic acid (DHA) is a long-chain, highly unsaturated omega-3 (n-3) fatty acid. It has a structure that gives it unique physical and functional properties. DHA is metabolically related to other n-3 fatty acids: it can be synthesised from the plant essential fatty acid α-linolenic acid (ALA). However, this pathway does not appear to be very efficient in many individuals, although the conversion of ALA to DHA is much better in young women than in young men. Furthermore, young infants may be more efficient converters of ALA to DHA than many adults, although the conversion rate is variable among infants. Many factors have been identified that affect the rate of conversion. The implication of poor conversion is that preformed DHA needs to be consumed. DHA is found in fairly high amounts in seafood, especially fatty fish, and in various forms of n-3 supplements. The amount of DHA in seafood and in supplements varies. Breast milk contains DHA. DHA is found esterified into complex lipids within the bloodstream, in adipose stores and in cell membranes. Its concentration in different compartments varies greatly. The brain and eye have high DHA contents compared to other organs. DHA is especially concentrated in the grey matter of the brain and in the rod outer segments of the retina. In the brain DHA is involved in neuronal signalling, while in the eye it is involved in the quality of vision. DHA is accumulated in the brain and eye late in pregnancy and in early infancy. A lower DHA content is linked to poorer cognitive development and visual function. DHA affects cell and tissue physiology and function through numerous mechanisms, including alterations in membrane structure and function, in membrane protein function, in cellular signalling and in lipid mediator production.
Docosahexaenoic Acid and Preterm Birth
Authors: Makrides Maria , Best Karen
Published: November 2016

Preterm birth accounts for more than 85% of all perinatal complications and deaths. There are many short- and long-term consequences of being born too soon. These infants often require intensive care and are at increased risk of early morbidities often with life-long sequelae. Approximately 50% of all preterm births have unknown or unclear causes, and there are no effective primary prevention strategies in widespread clinical use. Epidemiological studies have observed an increased length of gestation in populations with high fish consumption. These findings have led to randomised controlled trials of omega-3 (n-3) long-chain polyunsaturated fatty acid (LCPUFA) supplementation which show that these dietary agents may delay the timing of birth and may have value as a prophylactic intervention in some women. This review presents the available evidence and discusses the relationship between prenatal n-3 LCPUFA supplementation during pregnancy and the incidence of preterm birth.
Docosahexaenoic Acid and Neurodevelopmental Outcomes of Term Infants
Authors: Meldrum Suzanne , Simmer Karen
Published: November 2016

Docosahexaenoic acid (DHA), a long-chain polyunsaturated fatty acid, is essential for normal brain development. DHA is found predominantly in seafood, fish oil, breastmilk and supplemented formula. DHA intake in Western countries is often below recommendations. Observational studies have demonstrated an association between DHA intake in pregnancy and neurodevelopment of offspring but cannot fully adjust for confounding factors that influence child development. Randomised clinical trials of DHA supplementation during pregnancy and/or lactation, and of term infants, have not shown a consistent benefit nor harm on neurodevelopment of healthy children born at term. The evidence does not support DHA supplementation of healthy pregnant and lactating women, nor healthy infants.
Reference Ranges of Reticulocyte Haemoglobin Content in Preterm and Term Infants: A Retrospective Analysis
Authors: Lorenz Laila , Peter Andreas , Arand Jörg , Springer Fabian , Poets Christian F. , Franz Axel R.
Published: November 2016

Background: Despite iron supplementation, some preterm infants develop iron deficiency (ID). The optimal iron status parameter for early detection of ID has yet to be determined. Objective: To establish reference ranges for reticulocyte haemoglobin content (Ret-He) in preterm and term infants and to identify confounding factors. Methods: Retrospective analyses of Ret-He and complete blood count in infants with a clinically indicated blood sample obtained within 24 h after birth. Results: Mean (SD) Ret-He was 30.7 (3.0) pg in very preterm infants with a gestational age (GA) of <30 weeks (n = 55), 31.2 (2.6) pg in moderately preterm infants (GA 30-36 weeks, n = 241) and 32.0 (3.2) pg in term infants (GA ≥37 weeks, n = 216). The 2.5th percentile of Ret-He across all GA groups was 25 pg, with a weak correlation between Ret-He and GA (r = 0.18). Moreover, only weak/no correlations were found between Ret-He and C-reactive protein (r = 0.18), interleukin 6 (IL-6) (r = 0.03) and umbilical artery pH (r = -0.07). There was a slight variation in Ret-He with mode of delivery [normal vaginal delivery: 32.3 (3.2) pg, secondary caesarean section (CS): 31.4 (3.0) pg, instrumental delivery: 31.3 (2.7) pg and elective CS: 31.2 (2.8) pg]. Conclusion: GA at birth has a negligible impact on Ret-He, and the lower limit of the normal reference range in newborns within 24 h after birth can be set to 25 pg. Moreover, Ret-He seems to be a robust parameter which is not influenced by perinatal factors within the first 24 h after birth.
Hypoxic-Ischemic Encephalopathy-Associated Liver Fatty Degeneration and the Effects of Therapeutic Hypothermia in Newborn Piglets
Authors: Kubo Hiroyuki , Shimono Ryuichi , Nakamura Shinji , Koyano Kosuke , Jinnai Wataru , Yamato Satoshi , Yasuda Saneyuki , Nakamura Makoto , Tanaka Aya , Fujii Takayuki , Kanenishi Kenji , Chiba Yoichi , Miki Takanori , Kusaka Takashi , Ueno Masaki
Published: November 2016

Background: Although liver can be injured under the hypoxic-ischemic encephalopathy (HIE) condition, there is currently no histopathological evidence. Therapeutic hypothermia is used to protect the brain; however, the therapeutic potential for concomitant liver injury is unknown. Objectives: This study aimed to histopathologically prove HIE-associated liver injury and to investigate the influence of therapeutic hypothermia in a newborn piglet HIE model. Methods: Eighteen newborn piglets were divided into 3 groups: control (n = 4), HIE (n = 8), and therapeutic hypothermia (n = 6) groups. The hypoxic insult was induced by decreasing the fraction of inspiratory oxygen from 21 to 2-4% over 40 min while monitoring cerebral blood volume and cerebral hemoglobin oxygen saturation. For therapeutic hypothermia, whole-body cooling at 33-34°C was administered for 24 h after the hypoxic insult. We hematologically and histopathologically investigated the liver injury in all groups. Results: Alanine transaminase and lactate dehydrogenase levels in the HIE group were significantly elevated compared with those in the control group. Micro-lipid droplet accumulation in the periportal zone, but not in the perivenous zone, was significantly greater in the HIE group than in the control group and significantly smaller in the therapeutic hypothermia group than in the HIE group. Conclusions: We demonstrated that micro-lipid droplet accumulation in the cytoplasm of hepatocytes in the periportal zone occurs under the HIE condition and that this accumulation is suppressed by therapeutic hypothermia.
Prevention of Recurrent Bladder Tumors after Nephroureterectomy for Primary Upper Urinary Tract Urothelial Carcinoma: An Individualized Selection Strategy Based on Patient Risk Stratification Is Needed
Authors: Long Xingbo , Qi Lin , Zu Xiongbing , Li Yuan , He Wei , Pi Shuohuang , Chen Xiang , Zhou Kebing , Hu Xiheng , Chen Minfeng
Published: November 2016

Objective: To evaluate the efficacies of different chemotherapy regimens in different risk sub-groups for the prevention of bladder recurrence after radical nephroureterectomy (RNU). Materials and Methods: Between 2004 and 2012, we recruited 685 patients who underwent RNU for upper tract urothelial carcinoma at 4 Chinese institutions. We assessed whether the type of intravesical chemotherapy regimen affected the bladder recurrence rate in patients with different risk levels after RNU. Results: For all patients, the bladder recurrence rate was lower with intravesical chemotherapy than without, but no significant differences were found between the 2 intravesical chemotherapy regimens (single dose or relatively long-term therapy). We used multivariate analysis to define the risk factors for bladder recurrence and stratified patients into low-, intermediate-, and high-risk sub-groups accordingly. The bladder recurrence rate in the low-risk patients was not significantly different between patients with or without intravesical chemotherapy. However, in the intermediate-risk and high-risk patients, the rate was greater in patients without intravesical chemotherapy than in patients with. Furthermore, the rate was not significantly different between the 2 intravesical chemotherapy regimens in the intermediate-risk patients, while the efficacy of the relatively long-term regimen in high-risk patients remains unclear. Conclusions: Our study showed that an individualized strategy based on patient risk stratification is needed.