News Article | October 26, 2016
An international team comprising engineers, mathematicians and doctors has applied a technique used for detecting damage in underwater marine structures to identify cancerous cells in breast cancer histopathology images. Their multidisciplinary breakthrough, which has the potential to automate the screening of images and improve the detection rate, has been published in leading journal, PLOS ONE. The article is here: http://journals. Breast cancer is the most prevalent form of cancer for women worldwide. Current breast cancer clinical practice and treatment mainly relies on the evaluation of the disease's prognosis using the Bloom-Richardson grading system. The necessary scoring is based on a pathologist's visual examination of a tissue biopsy specimen under microscope, but different pathologists may assign different grades to the same specimens. However, the advent of digital pathology and fast digital slide scanners has opened the possibility of automating the prognosis by applying image-processing methods. While this undoubtedly represents progress, image-processing methods have struggled to analyse high-grade breast cancer cells as these cells are often clustered together and have vague boundaries, which makes successful detection extremely challenging. But the new method has seemingly overcome that task, according to Assistant Professor in Civil Engineering at Trinity College Dublin, Bidisha Ghosh. She said: "This unique research group could draw on a broad and deep knowledge base. Experts in numerical methods and image-processing liaised with medical pathologists, who were able to offer expert insight and could tell us precisely what information was of value to them. It is an excellent example of how multidisciplinary research collaborations can address important societal issues." Professor Joy John Mammen, Head of Department of Transfusion Medicine & Immunohaematology from the Christian Medical College, Vellore, India, said: "Detection of cancerous nuclei in high-grade breast cancer images is quite challenging and this work may be considered as a first step towards automating the prognosis." The proposed technique, previously used for detecting damaged surface areas on underwater marine structures such as bridge piers, off-shore wind turbine platforms and pipe-lines was applied to histopathology images of breast cells. The researchers considered the likelihood of every point in a histopathology image either being near a cell centre or a cell boundary. Using a belief propagation algorithm, the most suitable cell boundaries were then traced out. This technique was developed in conjunction with mathematicians in Madras Christian College, India. Lead author, Dr Maqlin Paramanandam, said: "The potential for this technology is very exciting and we are delighted that this international and inter-disciplinary team has worked so well at tackling a real bottle-neck in automating the diagnosis of breast cancer using histopathology images." Dr Michael O'Byrne, who also worked in University College Cork during this project, added: "Coming from a civil engineering background where most of our image-processing tools were designed to assess structural damage, it was nice to discover some cross-over applications and find areas where we could lend our expertise. We all found it particularly rewarding to contribute towards breast cancer research." The study was supported by a Science Foundation Ireland - International Strategic Cooperation Award.
News Article | December 7, 2016
BOSTON - A personalized cancer vaccine markedly improved outcomes for patients suffering from acute myeloid leukemia (AML), a potentially lethal blood cancer, in a clinical trial led by investigators at Beth Israel Deaconess Medical Center (BIDMC). The product of a long-term collaboration among investigators at the Cancer Center at BIDMC and Dana-Farber Cancer Institute, the vaccine stimulated powerful immune responses against AML cells and resulted in protection from relapse in a majority of patients, the team of researchers reported today in Science Translational Medicine. "Immunotherapy strategies leverage the body's own defense systems to fight cancer cells," said senior author David Avigan, MD, Chief, Section of Hematological Malignancies and Director of the Cancer Vaccine Program at the BIDMC Cancer Center and Professor of Medicine at Harvard Medical School. "By creating a personalized vaccine, we use the power of the immune system to selectively target each patient's cancer and avoid the side effects of chemotherapy." Patients with AML may achieve remission following standard chemotherapy, yet relapse is common, and most patients ultimately succumb to the disease. In this study, the team of collaborators from BIDMC and Dana-Farber generated personalized vaccines for 17 patients with AML who were in remission after undergoing standard chemotherapy. Despite an average age of 63, more than 70 percent of trial participants remained in remission at an average follow-up period of more than four years. After receiving a series of injections of the vaccine, patients demonstrated an increase in the number of leukemia-specific T cells in the blood and bone marrow. T cells are immune cells critical to the body's ability to recognize and remember pathogens like viruses, or in this case, cancer cells. Present only in low numbers prior to vaccination, T cells recognizing AML cells were expanded after vaccination, potentially providing long-term protection against the leukemia. "With the vaccine, we use the immune system to target the whole tumor including cells that may be resistant to chemotherapy," stated lead author Jacalyn Rosenblatt, MD, Co-Director of the Cancer Vaccine Program at the BIDMC Cancer Center and Associate Professor of Medicine at Harvard Medical School. "We were really excited to see that the vaccine generated a broad and durable immune response without significant side effects." This vaccine platform has been the product of collaboration among BIDMC and Dana-Farber investigators, including the initial seminal work done by Donald W. Kufe, MD, Distinguished Physician at Dana-Farber, subsequent development and clinical translation by Kufe, Rosenblatt and Avigan, and the contributions of clinical investigators including Richard Stone, MD, Chief of the Adult Leukemia Program at Dana-Farber, and Lynne Uhl, MD, Director of Transfusion Medicine at BIDMC. "The development of this personalized vaccine by our team was based on the premise that effective treatment of established cancers would require the induction of immunity against multiple antigens, including neoantigens, specifically expressed by the patient's own cancer cells," stated co-author Donald W. Kufe, MD. Based on these encouraging results, researchers are also testing this vaccine approach in other types of cancers. Avigan and colleagues are leading a national study to test the effectiveness of the vaccine in patients with multiple myeloma, another common blood cancer. Conducted under the auspices of the NIH-sponsored Clinical Trials Network, this first-of-its-kind endeavor brings together 15 leading cancer centers. Of note, the unique research effort takes an open-source approach, in which participating sites were trained in vaccine production at BIDMC and will work together to bring this therapy to patients nationwide. Study coauthors also include Lynne Uhl, MD; Robin Joyce, MD; James D. Levine, MD; Jon Arnason, MD; Malgorzata McMasters, MD; Katarina Luptakova, MD; Salvia Jain, MD; Jeffrey I Zwicker, MD; Ayad Hamdan, MD; Vassiliki Boussiotis, MD, PhD; Poorvi Somaiya Dutt, MS; Emma Logan, BSN; Mary Paty Bryant; Dina Stroopinsky, PhD; Kristen Palmer, MA; Max Coll; Abigail Washington; and Leandra Cole, all of BIDMC. Co-authors also include Richard M. Stone, MD; David P. Steensma, MD; Daniel J. deAngelo, MD, PhD; and Ilene Galinsky, MSN, all of Dana-Farber. This work was supported by grants from the National Institutes of Health/National Cancer Institute (NIH/NCI R21CA149987-02) and from the Leukemia and Lymphoma Society Translational Research Program. Additional support for early vaccine work was provided by the Louis B. Mayer Foundation, and the Barbara and James Sadowsky Fund.
News Article | November 15, 2016
UMass Memorial Health Care in Worcester, MA, the largest healthcare system in central Massachusetts, has recently contracted with Mediware Information Systems, Inc. to implement HCLL™ Transfusion, HCLL™ Analytics, LifeTrak®, and Transtem™. The best-of-breed applications are the core software solutions comprising Mediware’s Blood Management Technologies division. HCLL Transfusion, Mediware’s 510(k)-cleared transfusion management system utilized by hundreds of prestigious hospitals and the Department of Defense (DoD), features robust patient safety protocols and multi-site and multi-facility inventory management capabilities. HCLL Analytics adds performance management tools to the transfusion management system. These applications will be implemented across the UMass Memorial Health Care system, which includes four community hospitals and a Level 1 trauma center with advanced emergency services. Mediware’s LifeTrak donor software, also with 510(k) clearance, and its Transtem cellular therapy software will be used at UMass Memorial Medical Center, which is the clinical partner of UMass Medical School. LifeTrak, relied on by the nation’s most renowned blood centers, hospitals, and the DoD, will reliably manage donor information and eligibility, product manufacturing, and distribution processes. Transtem, an innovative cellular therapy solution, will provide end-to-end process management from cell or material collection through all phases of manufacturing and, ultimately, product release for use in clinical therapy. “With all four systems in place, we are confident that we can continue to maintain our commitment to improving the health of people in central Massachusetts through excellent care, comprehensive health services, and cutting-edge research,” said Dr. Robert Weinstein, chief of UMass Memorial Medical Center’s Division of Transfusion Medicine. “Mediware is pleased to partner with the UMass Memorial Health Care System by providing our innovative, stand-alone applications and supporting seamless integration across the system’s multiple facilities,” stated Thomas Mann, Mediware’s president and CEO. About Mediware Information Systems Mediware is a leading supplier of software for healthcare and human service providers and payers. For more than 30 years, we have equipped the nation’s blood banks with the most comprehensive solutions on the market. Today, more than 1,500 hospitals and blood centers rely on our blood donor and transfusion management applications for fast, error-free collection, supply and usage. Our portfolio of solutions also includes human services, cellular therapy, home care, medication management, rehabilitation, and respiratory therapy, all developed by our 600+ subject matter experts who understand business and care processes in specialized acute, non-acute, and community-based care settings. For more information about Mediware products and services, visit http://www.mediware.com. About UMass Memorial Health Care UMass Memorial Health Care is the largest healthcare system in central Massachusetts. The system includes four hospitals and is the clinical partner of UMass Medical School, which provides access to the latest technology, research, and clinical trials. The hospitals are accredited by the Joint Commission or by Det Norske Veritas (DNV), a risk management company that helps improve healthcare services. In addition to fully equipped medical centers, the system also includes home health and hospice programs, behavioral health programs, and community-based physician practices.
News Article | December 14, 2016
The International Association of HealthCare Professionals is pleased to welcome Yvette C. Tanhehco, MD, PhD, MS, to their prestigious organization with her upcoming publication in The Leading Physicians of the World. Dr. Tanhehco is a highly-trained and qualified Clinical Pathologist with an extensive expertise in all facets of her work, especially Transfusion Medicine. Dr. Tanhehco has been in practice for over four years and is currently serving as Attending Physician in Transfusion Medicine, Director of Apheresis and Cellular Therapy, and Assistant Director of Transfusion Medicine at NewYork-Presbyterian Hospital/Columbia University Irving Medical Center in New York City, New York. Dr. Tanhehco’s career in medicine began in 2008 when she graduated with her Medical Degree from the University of Pittsburgh, followed by her residency training in Clinical Pathology and fellowship training in Transfusion Medicine at the University of Pennsylvania. In addition to her Medical Degree, Dr. Tanhehco also holds a PhD in Viral Oncology from Johns Hopkins University and a Master of Science Degree in Translational Research obtained at the University of Pennsylvania. Furthermore, Dr. Tanhehco is double board certified in Clinical Pathology and in Transfusion Medicine. To keep up to date with the latest advances and developments in her field, Dr. Tanhehco maintains a professional membership with the American Association of Blood Banks, the American Society for Apheresis, and the College of American Pathologists. Dr. Tanhehco is interested in studying thrombotic disorders such as thrombotic thrombocytopenic purpura and heparin induced thrombocytopenia, especially the role of plasma exchange for the treatment of these disorders. She is also interested in studying novel peripheral blood stem cell mobilization agents and understanding the factor that contribute to a successful mobilization and collection. She attributes her great success in her career to her hard work, personal values, excellent training, her mentors along the way, the right opportunities, and a supportive family. When she is not assisting patients, Dr. Tanhehco enjoys swimming, reading books, watching movies, and traveling. Learn more about Dr. Tanhehco by reading her upcoming publication in The Leading Physicians of the World. FindaTopDoc.com is a hub for all things medicine, featuring detailed descriptions of medical professionals across all areas of expertise, and information on thousands of healthcare topics. Each month, millions of patients use FindaTopDoc to find a doctor nearby and instantly book an appointment online or create a review. FindaTopDoc.com features each doctor’s full professional biography highlighting their achievements, experience, patient reviews, and areas of expertise. A leading provider of valuable health information that helps empower patient and doctor alike, FindaTopDoc enables readers to live a happier and healthier life. For more information about FindaTopDoc, visit:http://www.findatopdoc.com
News Article | March 9, 2016
LAP-tTA and TRE-MYC mice were previously described and MYC expression in the liver was activated by removing doxycycline treatment (100 μg ml−1) from the drinking water of 4-week-old double transgenic mice for both TRE-MYC and LAP-tTA as previously described9, 13. C57BL/6 mice were obtained from NCI Frederick. Chemically induced HCC was established by intraperitoneal injection of diethylnitrosoamine (DEN) (Sigma) into 2-week-old male pups at a dose of 20 μg g−1 body weight13. Twelve-week-old male B6.Cg-Lepob/J (ob/ob) mice or wild-type control mice were obtained from Charles River. Foxp3–GFP mice were previously described31. NAFLD was induced by feeding mice with a methionine–choline-deficient (MCD) diet (catalogue number 960439, MP biomedical), a choline-deficient and amino-acid-defined (CDAA) diet (catalogue number 518753, Dyets) or a high-fat diet (catalogue number F3282, Bio Serv) for the indicated time10, 11, 32. The MCD diet was supplied with corn oil (10%, w/w), and no fish oil was added. Control diet was purchased from MP Biomedical (catalogue number 960441). Custom-made high- or low-linoleic-acid mouse diets were purchased from Research Diets. The modified diets were based on AIN-76A standard mouse diet, and are isocaloric (4.45 kcal g−1) and contained the same high-fat content (23%, w/w). Linoleic-acid-rich safflower oil and saturated fatty-acid-containing coconut oil were supplied at different ratios to yield 2% (w/w) for the low-linoleic-acid diet or 12% (w/w) for the high-linoleic-acid diet. C57BL/6 mice were fed with the high- or low-linoleic-acid diet for 4 weeks. MYC mice were injected i.p. with 50 μg CD4 antibody (clone GK1.5, BioXcell) every week for the indicated time period to deplete CD4+ T cells33. N-acetylcysteine (NAC) was given in drinking water (10 mg ml−1)34 for the indicated time period to prevent excess ROS production. Mitochondrial-specific antioxidant mitoTEMPO was purchased from Sigma. Mice received mitoTEMPO at a dose of 0.7 mg kg−1 per day25 by osmotic minipumps (ALZET). At the experimental end points, mice were killed. For flow cytometry analysis, single-cell suspensions were prepared from spleen, liver and blood as described previously. Red blood cells were lysed by ACK Lysis Buffer (Quality Biologicals). Parts of live tissue were fixed by 10% formaldehyde and subjected to H&E staining. Free fatty acids were purchased from Sigma. Lipid accumulation was detected by Oil Red O staining in frozen liver sections using the custom service of Histo Serv. Cells were surface-labelled with the indicated antibodies for 15 min at 4 °C. Flow cytometry was performed on BD FACSCalibur or BD LSRII platforms and results were analysed using FlowJo software version 18.104.22.168 (TreeStar). The following antibodies were used for flow cytometry analysis: anti-CD3-FITC (clone 17A2, BD Pharmingen), anti-CD4-PE (clone RM4–4, Biolegend), anti-CD4-APC (clone RM4–5, eBioscience), anti-CD8-Alexa Fluor 700 (clone 53–6.7 Biolegend), anti-CD45, anti-CD44-PE (clone IM7, eBioscience), anti-CD62L-PerCP/Cy5.5 (MEL-14, Biolegend), anti-CD69-Pacific blue (clone H1.2F3, Biolegend), PBS57/CD1d-tetramer-APC (NIH core facility). To determine cytokine production, cells were stimulated with PMA and ionomycine for 30 min, and then were fixed and permeabilized using cytofix/cytoperm kit (BD Pharmingen) followed by anti-IFN-γ-PE (clone XMG1.2, BD Pharmingen), anti-IL-17-PerCP/Cy5.5 (clone TC11-18H10.1, Biolegend) staining. Cell death and apoptosis were detected with annexin V-PE (BD Pharmingen) and 7-AAD (BD Pharmingen) staining according to the manufacturer’s instructions. Intrahepatic CD4+ lymphocytes were gated on the CD3hiCD4+ population from total live hepatic infiltrating mononuclear cells. Absolute numbers were calculated by multiplying frequencies obtained from flow by total live mononuclear cell count, then divided by liver weight. The antibodies used for human peripheral blood mononuclear cell (PBMC) staining are the following: anti-CD3-PE (clone SK7, BD Pharmingen), anti-CD4-FITC (clone RPA-T4, BD Pharmingen), anti-CD8-APC (clone RPA-T8, BD Pharmingen). Murine T assays were performed as described31. Briefly, liver T cells were isolated as CD4+GFP+ by flow-cytometry-assisted cell sorting from Foxp3–GFP mice kept on an MCD or control diet for 4 weeks. CD4+GFP− T effector (T ) cells (5 × 104) were stimulated for 72 h in the presence of irradiated T-depleted splenocytes (5 × 104) plus CD3ε monoclonal antibody (1 μg ml−1), with or without T cells added at different ratios. 3H-Thymidine was added to the culture for the last 6 h and incorporated radioactivity was measured. Freshly isolated splenocytes from MYC-ON MCD mice were incubated with 5 μg ml−1 of mouse α-fetoprotein protein (MyBioSource) for 24 h. Golgiplug was added for the last 6 h. Then, cells were fixed and permeabilized using cytofix/cytoperm kit (BD Pharmingen) followed by anti-IFN-γ-PE (clone XMG1.2, BD Pharmingen) staining. Primary mouse hepatocytes were isolated from MYC mice and cultured according to a previous report35. Briefly, mice were anaesthetized and the portal vein was cannulated under aseptic conditions. The livers were perfused with EGTA solution (5.4 mM KCl, 0.44 mM KH PO , 140 mM NaCl, 0.34 mM Na HPO , 0.5 mM EGTA, 25 mM Tricine, pH 7.2) and Gey’s balanced salt solution (Sigma), and digested with 0.075% collagenase solution. The isolated mouse hepatocytes were then cultured with complete RPMI media in collagen-I-coated plates. Hepatic fatty acid composition was measured at LIPID MAPS lipidomics core at the University of California (San Diego) using an esterified and non-esterified (total) fatty acid panel. Briefly, liver tissues were homogenized and lipid fraction was extracted using a modified Bligh Dyer liquid/liquid extraction method. The lipids were saponified and the hydrolysed fatty acids were extracted using a liquid/liquid method. The extracted fatty acids were derivatized using pentaflourylbenzylbromine (PFBB) and analysed by gas chromatography (GC) using an Agilent GC/mass spectrometry (MS) ChemStation. Individual analytes were monitored using selective ion monitoring (SIM). Analytes were monitored by peak area and quantified using the isotope dilution method using a deuterated internal standard and a standard curve. Isolated primary hepatocytes from MYC mice fed with MCD or control diet were cultured in complete RPMI for 24 h. Supernatant were harvested and FFAs were identified by GC/MS. Splenocytes from MYC mice were cultured with or without 50 μM C18:2 for 24 h. CD4+ and CD8+ T lymphocytes were sorted and total RNA was extracted using miRNeasy mini kit (Qiagen). Array analysis was performed in the Department of Transfusion Medicine, clinical centre at NIH. Mouse gene 2.0 ST array (Affymetrix) was used and performed according to the manufacturer’s instruction. Data were log-transformed (base 2) for subsequent statistical analysis. The Partek Genomic Suite 6.4 was used for the identification of differentially expressed transcripts. The Ingenuity Pathway Analysis tool (http://www.ingenuity.com) was used for analysis of functional pathways. RNA was extracted from frozen tissues with RNeasyMini Kit (Qiagen). Complementary DNA was synthesized by iScriptcDNA synthesis kit (BioRad). Sequence of primers used for quantitative RT–PCR can be obtained from the authors. The reactions were run in triplicates using iQSYBR green supermix kit (BioRad). The results were normalized to endogenous GAPDH expression levels. CD4+ T lymphocytes were isolated from the spleen of MYC mice by negative autoMACS selection using a CD4+ T lymphocytes isolation kit (Miltenyi Biotec) or flow cytometry cell sorting. Human CD4+ T lymphocytes were prepared from PBMCs by autoMACS using a CD4+ T lymphocytes isolation kit (Miltenyi Biotec). The purity of CD4+ T lymphocytes was above 90% after autoMACS separation and above 95% after flow cytometry cell sorting. C16:0, C18:0, C18:1,and C18:2 were purchased from Sigma. Fatty acids were dissolved in DMEM with 2% fatty-acid-free bovine serum albumin (BSA; Sigma, catalogue number A8806) after solvent was evaporated, then followed by two rounds of vortexing and 30 s of sonication. Isolated CD4+ T lymphocytes or splenocytes were incubated with different fatty acids or conditioned medium from hepatocyte culture for 3 days. Unless specifically described, fatty acids were used at 50 μM concentration. For fatty acid depletion, active charcoal (catalogue number C-170, Fisher) was used as described before36. Briefly, 0.5 g of active charcoal was added into every 10 ml of conditioned medium. Then pH was lowered to 3.0 by addition of 0.2 N HCl. The solution was rotated at 4 °C for 2 h. Charcoal was then removed by centrifugation, and the clarified solution was brought back to pH 7.0 by addition of 0.2 N NaOH. NAC (10 mM), catalase (1,000 U ml−1) or mitoTEMPO (10 μM) was used to inhibit ROS production, mitochondrial ROS levels were determined by mitoSOX staining 24 h after treatment, cell death and apoptosis were measured by annexin V and 7-AAD staining 3 days after treatment. Caspase activity assay was measured by caspase-Glo 3/7 assay kit (Promega) according to the manufacturer’s protocol. Fresh prepared liver-infiltrating mononuclear cells were washed and resuspended in 500 μl of BODIPY 493/503 at 0.5 μg ml−1 in PBS. Cells were stained for 15 min at room temperature. Then cells were subjected to flow cytometry analysis. Two pZIP lentiviral shRNA vectors targeting human CPT1a and a control vector (NT#4) were purchased from TransOMIC Technologies. Lentivirus was packed in 293T cells. Jurkat cells were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ), and no authentication test was performed by us. Cells were cultured in complete RPMI medium and were tested to be mycoplasma free. Jurkat cells were infected with shRNA lentivirus. Puromycin was added to eliminate non-transduced cells. Doxycycline (100 ng ml−1) was added to induce shRNA and GFP expression for 3 days. Efficiency of shRNAs was confirmed by western blot. Jurkat cells with CPT1a knockdown were treated with 200 μM C18:2 for 24 h. Mitochondrial ROS production and cell survival were measured in GFP+-transduced cells. Fatty acid oxidation was measured according to a previous publication37. 1-14C-C18:2 and 1-14C-C16:0 were purchased from PerkinElmer. Briefly, isolated CD4+ or CD8+ T lymphocytes were pretreated with C18:2 or kept in regular media. After 24 h, cell media was changed to media containing 50 μM cold C18:2 plus 1 μCi 1-14C-C18:2 per ml or 50 μM cold C16:0 plus 1 μCi 1-14C-C16:0 per ml. After 2 h, medium was removed and mixed with concentrated perchloric acid (final concentration 0.3 M) plus BSA (final concentration 2%) to precipitate the radiolabelled fatty acids. Samples were vortexed and centrifuged (10,000g for 10 min). Radioactivity was determined in the supernatant to measure water-soluble β-oxidation products. Mitochondrial membrane potential was measured by TMRM (ImmunoChemistry Technologies) staining according to the manufacturer’s protocol. Briefly, cells were kept in culture medium with 100 nM of TMRM for 20 min in a CO incubator at 37 °C. After washing twice, cells were processed to flow cytometry analysis. Mitochondria-associated superoxide was detected by mitoSOX (Life Technologies) staining according to the manufacturer’s protocol. Briefly, cells were first subjected to surface marker staining. Then cells were stained with 2.5 μM mitoSOX for 30 min in a CO incubator at 37 °C. After washing twice, cells were processed for flow cytometry analysis. OCR was measured using an XFe96 Extracellular Flux Analyzer (Seahorse Bioscience) as previously described38. AutoMACS-sorted mouse CD4+ and CD8+ T lymphocytes were attached to XFe96 cell culture plates using Cell-Tak (BD Bioscience) in RPMI media with 11 mM glucose. Cells were activated with 1:1 CD3:CD28 beads (Miltenyi BioTech) and vehicle or 50 μM C18:2 was added. Twenty-four hours after activation, cells were incubated in serum-free XF Base Media (Seahorse Bioscience) supplemented with 10 mM glucose, 2 mM pyruvate and 2 μM glutamine, pH 7.4, along with 50 μM C18:2 if previously present, for 30 min at 37 °C in a CO -free cell culture incubator before beginning the assay. Five consecutive measurements, each representing the mean of 8 wells, were obtained at baseline and after sequential addition of 1.25 μM oligomycin, 0.25 μM trifluorocarbonylcyanide phenylhydrazone (FCCP), and 1 μM each of rotenone and antimycin A (all drugs from Seahorse Bioscience). OCR values were normalized to cell number as measured by the CyQUANT Cell Proliferation Assay Kit (Life Technologies). Human liver samples were stained as previously described8. For immunostaining, formalin-fixed, paraffin-embedded human liver tissue samples were retrieved from the archives of the Institute of Surgical Pathology, University Hospital Zurich. Fibrosis grade was analysed for NASH according to NAFLD activity score (NAS)39 and for others according to METAVIR score40. The study was approved by the local ethics committee (Kantonale Ethikkommission Zürich, application number KEK-ZH-Nr. 2013-0382). Human PBMCs from healthy donors were obtained on an NIH-approved protocol and prepared as described previously41. Informed consent was obtained from all subjects. The sample sizes for animal studies were guided by a previous study in our laboratory in which the same MYC transgenic mouse stain was used. No animals were excluded. Neither randomization nor blinding were done during the in vivo study. However, mice from the same littermates were evenly distributed into control or treatment groups whenever possible. The sample size for the patient studies was guided by a recent publication also studying NASH-induced HCC, but focused on different aspects8. Statistical analysis was performed with GraphPad Prism 6 (GraphPad Software). Significance of the difference between groups was calculated by Student’s unpaired t-test, one-way or two-way ANOVA (Tukey’s and Bonferroni’s multiple comparison test). Welch’s corrections were used when variances between groups were unequal. P < 0.05 was considered as statistically significant.
Armand-Ugon R.,Sant Joan Despi Moises Broggi Hospital |
Cheong T.,Francisco Palau Community Hospital |
Matapandewu G.,Francisco Palau Community Hospital |
Rojo-Sanchis A.,Sant Joan Despi Moises Broggi Hospital |
And 2 more authors.
Journal of Women's Health | Year: 2011
Background: Postpartum hemorrhage may lead to maternal morbidity and mortality, increases risks of transfusion, and incurs costs. We report on the feasibility and efficacy of in-hospital intravenous (IV) iron for treating postoperative anemia at Mtengo wa Nthenga, Malawi. Patients and methods: Twenty-eight consecutive women undergoing surgery for complicated pregnancy or complicated childbirth entered the study. Patients with hemoglobin (Hb) <10g/dL on postoperative day 1 (n=14) received IV iron sucrose (200mg/day, 3 consecutive days), and those with Hb ≥10g/dL (n=14) received oral iron (ferrous sulfate, 256mg/day). In-hospital postoperative Hb increase and blood transfusion were recorded. Results: Mean changes in Hb from postoperative day 1 to postoperative day 7 were-0.6±1.2g/dL and 2.1±1.7g/dL, for the oral and IV iron groups, respectively (p=0.001). No side effect was seen with IV iron. Only 1 of 4 women receiving allogeneic blood was transfused after the initiation of IV iron treatment. Conclusions: Our results suggest that IV iron sucrose is an effective drug for treating puerperal anemia, leading to a rapid recovery of Hb levels. The current availability of generic iron sucrose preparations, with considerably lower acquisition costs, may facilitate in-hospital access to this treatment option in low-resource countries. © 2011, Mary Ann Liebert, Inc.
Franchini M.,Transfusion Medicine |
Favaloro E.J.,Westmead Hospital |
Lippi G.,Academic Hospital of Parma
Seminars in Thrombosis and Hemostasis | Year: 2015
The mainstay of treatment of inherited coagulation disorders is based on the infusion of the deficient clotting factor, when available. Significant advances have been made over the past two decades in the production and availability of factor replacement products. In spite of such progression, several issue are still unsolved, the most important being the need for frequent factor concentrate infusions and the development of inhibitory alloantibodies. To overcome these important limitations, several newer hemostatic agents with an extended half-life are at an advanced stage of clinical development. After a brief overview of hemostasis, this narrative review summarizes the current knowledge on the most promising novel products for hemostasis. The current status of gene therapy for hemophilia, the only therapeutic option to definitively cure this inherited bleeding disorder, is also concisely discussed. © 2015 by Thieme Medical Publishers, Inc.
Armenian S.H.,Outcomes Research |
Ding Y.,Outcomes Research |
Mills G.,Outcomes Research |
Sun C.,Outcomes Research |
And 7 more authors.
British Journal of Haematology | Year: 2013
Summary: Haematopoietic cell transplantation (HCT) survivors are at increased risk for developing congestive heart failure (CHF), primarily due to pre-HCT exposure to anthracyclines. We examined the association between the development of CHF after HCT and polymorphisms in 16 candidate genes involved in anthracycline metabolism, iron homeostasis, anti-oxidant defence, and myocardial remodelling. A nested case-control study design was used. Cases (post-HCT CHF) were identified from 2950 patients who underwent HCT between 1988 and 2007 at City of Hope and had survived ≥1 year. This cohort formed the sampling frame for selecting controls (without CHF) matched on: age, race/ethnicity, cumulative anthracycline exposure, stem cell source (allogeneic, autologous), and length of follow-up. Seventy-seven cases with pre-HCT germline DNA and 178 controls were genotyped. Multivariate analysis revealed that the odds of CHF was higher in females [Odds Ratio (OR) = 2·9, P < 0·01], individuals with pre-HCT chest radiation (OR = 4·7, P = 0·05), hypertension (OR = 2·9, P = 0·01), and with variants of genes coding for the NAD(P)H-oxidase subunit RAC2 (rs13058338, 7508T→A; OR = 2·8, P < 0·01), HFE (rs1799945, 63C→G; OR = 2·5, P = 0·05) or the doxorubicin efflux transporter ABCC2 (rs8187710, 1515G→A; OR = 4·3, P < 0·01). A combined (clinical and genetic) CHF predictive model performed better [area under the curve (AUC), 0·79] than the genetic (AUC = 0·67) or the clinical (AUC = 0·69) models alone. © 2013 John Wiley & Sons Ltd.
Ius F.,Hannover Medical School |
Sommer W.,Hannover Medical School |
Tudorache I.,Hannover Medical School |
Kuhn C.,Hannover Medical School |
And 12 more authors.
Journal of Heart and Lung Transplantation | Year: 2015
Objective De novo donor-specific anti-human leukocyte antigen antibodies develop in a high proportion of lung transplant recipients early after lung transplantation. We recently showed that de novo donor-specific antibodies (DSA) occurrence is associated with significantly increased mortality. Here, we studied the efficacy of a preemptive treatment protocol.Methods A retrospective observational study was conducted on all lung transplantations at Hanover Medical School between January 2009 and May 2013.Results Among the 500 transplant recipients, early DSA developed in 86 (17%). Of these, 56 patients (65%; Group A) received therapeutic plasma exchange, and 30 patients (35%; Group B) did not. Among Group A patients, 51 also received rituximab. Between groups, there was no statistically significant difference in mortality, incidence of pulsed steroid therapies, rejections diagnosed by biopsy specimen, incidence of bronchitis obliterans syndrome (BOS), or infections requiring hospitalization at 1 year and 3 years. Also, there were no statistically significant differences after matching 21 Group A with 21 Group B patients through propensity score analysis. Significantly more Group A patients (65%) than Group B patients (34%) cleared DSA at hospital discharge (p = 0.01). At the last control after transplantation (median, 14 months; interquartile range, 5-24 months), 11 Group A (22%) and 9 Group B patients (33%) still showed DSA (p = 0.28).Conclusions Preemptive treatment with therapeutic plasma exchange and rituximab led to improved elimination of DSA early after lung transplantation (p = 0.01). However, spontaneous elimination in untreated Group B patients also occurred frequently. This treatment protocol was not associated with significantly improved outcome. © 2015 International Society for Heart and Lung Transplantation.
Holgersson J.,Transfusion Medicine |
Rydberg L.,Transfusion Medicine |
Breimer M.E.,Gothenburg University
International Reviews of Immunology | Year: 2014
In recent years ABO incompatible kidney transplantation (KTx) has become a more or less clinical routine procedure with graft and patient survival similar to those of ABO compatible transplants. Antigen-specific immunoadsorption (IA) for anti-A and anti-B antibody removal constitutes in many centers an important part of the treatment protocol. ABO antibody titration by hemagglutination is guiding the treatment; both if the recipient can be transplanted as well as in cases of suspected rejections if antibody removal should be performed. Despite the overall success of ABO incompatible KTx, there is still room for improvements and an extension of the technology to include other solid organs. Based on an increased understanding of the structural complexity and tissue distribution of ABH antigens and the fine epitope specificity of the ABO antibody repertoire, improved IA matrices and ABO antibody diagnostics should be developed. Furthermore, understanding the molecular mechanisms behind accommodation of ABO incompatible renal allografts could make it possible to induce long-term allograft acceptance also in human leukocyte antigen (HLA) sensitized recipients and, perhaps, also make clinical xenotransplantation possible. © 2014 Informa Healthcare USA, Inc.