Medical Anatomy Commons^™

How The Manipulation Of The Ras Homolog Enriched In Striatum Alters The Behavioral And Molecular Progression Of Huntington’S Disease, Franklin A. Lee

University of New Orleans Theses and Dissertations

Huntington’s disease is an incurable, progressive neurological disorder characterized by loss of motor control, psychiatric dysfunction, and eventual dystonia leading to death. Despite the fact that this disorder is caused by a mutation in one single gene, there is no cure. The mutant Huntingtin (mHtt) protein is expressed ubiquitously throughout the brain but frank cell death is limited to the striatum. Recent work has suggested that Rhes, Ras homolog enriched in striatum, which is selectively expressed in the striatum, may play a role in Huntington’s disease neuropathology. In vitro studies have shown Rhes to be an E3 ligase for the …

Progtar: A Database Of Prognostically Inversely Correlated Mirnas And Genes (Pics) In Multiple Cancers, Chirayu Pankaj Goswami

Department of Pathology, Anatomy, and Cell Biology Posters

ABSTRACT

PROGTar is a database of Prognostically Inversely Correlated miRNA-mRNA pairs (PIC’s) in 23 cancer types. Partner miRNA and mRNA in a PIC show inverse correlation of expression and opposite hazards. We analyzed miRNA and mRNA expression data downloaded from The Cancer Genome Atlas (TCGA) in a 3 step approach to identify PICs in different cancer types. In first step we performed correlation analysis between miRNAs and mRNAs for each cancer type. This was followed by performing hazard analysis separately for miRNAs and mRNAs using expression data and survival related clinical variables. In the third step we merged the correlation …

Comprehensive Profiling Of Metaplastic Breast Carcinoma Reveals Frequent Over-Expression Of Pd-L1, Zoran Gatalica, Upasana Joneja, Md, Anatole Ghazalpour, Jeffrey Swensen, Rebecca Feldman, Fred Cai, Sting Chen, Nick Xiao, Sandeep Reddy, Semir Vranić

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

Background:

Metaplastic breast carcinoma (MBC) is a rare subtype of breast carcinoma less responsive to conventional chemotherapy relative to usual breast carcinomas such as ductal and lobular subtype. In molecular terms, MBC usually clusters with triple negative breast cancers (TNBC), but MBCs portray a worse prognosis in comparison with TNBC. Published studies investigating MBCs for specific biomarkers of therapy response are rare and limited by the methodological approaches.

Methods:

297 samples [MBC (n=75), triple-negative breast cancer of no-special-type (TNBC-NOS, n=106), HER2-positive breast cancers (n=32) and luminal breast cancers (n=84)] were profiled using direct sequencing analysis [Illumina MiSeq Next Generation Sequencing …

T(3;8)(Q26;Q24) With Myc Rearrangement In Acute Myeloid Leukemia: A Case Report, Vandi Ly, Md, Renu Bajaj, Phd, Jerald Z. Gong, Md, Zi-Xuan Wang, Phd, Stephen C Peiper, Md, Guldeep Uppal, Md

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Rearrangements of 3q26 have been described in 5% of de novo or therapy related acute myeloid leukemia, myelodysplastic syndrome (MDS), and blast phase of chronic myeloid leukemia. The most common translocations involving 3q26 are t(3;12)(q26;p13), t(3;21)(q26;q22), t(3;3)(q21;q26), t(2;3)(p15∼23;q26∼27) and rarely t(3;7)(q26;q21). However, t(3;8)(q26;q24) with or without monosomy 7 is a rare phenomenon and has been reported in only 10 patients so far. Hereby, we describe a 58 year old patient who was diagnosed with refractory anemia with multilineage dysplasia. Cytogenetic studies revealed monosomy 7. He was then lost to follow-up. A year later he was found to have worsening cytopenias …

Discovering Distinct Functional Modules Of Specific Cancer Types Using Protein-Protein Interaction Networks., Ru Shen, Xiaosheng Wang, Chittibabu Guda

Journal Articles: Genetics, Cell Biology & Anatomy

Background. The molecular profiles exhibited in different cancer types are very different; hence, discovering distinct functional modules associated with specific cancer types is very important to understand the distinct functions associated with them. Protein-protein interaction networks carry vital information about molecular interactions in cellular systems, and identification of functional modules (subgraphs) in these networks is one of the most important applications of biological network analysis. Results. In this study, we developed a new graph theory based method to identify distinct functional modules from nine different cancer protein-protein interaction networks. The method is composed of three major steps: (i) extracting modules …

Crispr/Cas9-Based Generation Of Knockdown Mice By Intronic Insertion Of Artificial Microrna Using Longer Single-Stranded Dna., Hiromi Miura, Channabasavaiah B. Gurumurthy, Takehito Sato, Masahiro Sato, Masato Ohtsuka

Journal Articles: Genetics, Cell Biology & Anatomy

Knockdown mouse models, where gene dosages can be modulated, provide valuable insights into gene function. Typically, such models are generated by embryonic stem (ES) cell-based targeted insertion, or pronuclear injection, of the knockdown expression cassette. However, these methods are associated with laborious and time-consuming steps, such as the generation of large constructs with elements needed for expression of a functional RNAi-cassette, ES-cell handling, or screening for mice with the desired knockdown effect. Here, we demonstrate that reliable knockdown models can be generated by targeted insertion of artificial microRNA (amiRNA) sequences into a specific locus in the genome [such as intronic …

Gonad: Genome-Editing Via Oviductal Nucleic Acids Delivery System: A Novel Microinjection Independent Genome Engineering Method In Mice., Gou Takahashi, Channabasavaiah B. Gurumurthy, Kenta Wada, Hiromi Miura, Masahiro Sato, Masato Ohtsuka

Journal Articles: Genetics, Cell Biology & Anatomy

Microinjection is considered the gold standard technique for delivery of nucleic acids (NAs; transgenes or genome editing tools such as CRISPR/Cas9 systems) into embryos, for creating genetically modified organisms. It requires sophisticated equipment as well as well-trained and highly skilled personnel to perform the micro-injection technique. Here, we describe a novel and simple microinjection-independent technique, called Genome-editing via Oviductal Nucleic Acids Delivery (GONAD). Using GONAD, we show that NAs (e.g., eGFP mRNA or Cas9 mRNA/sgRNAs) can be effectively delivered to pre-implantation embryos within the intact mouse oviduct by a simple electroporation method, and result in the desired genetic modification in …

Strategic Distribution Of The Mitochondrial Ca2+ Uniporter In Cardiac Mitochondria, Sergio De La Fuente, Caitlin Vail, Elorm J. Agra, Kira Holmstrom, Junhui Sun, Jyotsna Mishra, Toren Finkel, Elizabeth Murphy, Shey-Shing Sheu, György Csordás

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Background:

1) Control of the mitochondrial ATP production by SR-derived Ca²⁺ signals includes local, nanodomain Ca²⁺ transfer from ryanodine receptors (RyR2) to the mitochondrial matrix (excitation-bioenergetics coupling).

2) Ca²⁺ crosses the inner mitochondrial membrane (IMM) via the mtCU, a low-affinity Ca²⁺-activated Ca²⁺ channel complex.

3) The surface area of cardiac IMM is extensively enhanced by cristae folding; however, mitoplast patch clamp studies showed mtCU current density the lowest amongst a range of tissues (Fieni 2012. Nat Commun).

Low Level Lead Exposure Impairs Attentional Set Shifting Task Performance Depending Upon Sex And Developmental Periods Of Exposure, L.S. Neuwirth, D.W. Anderson, J. S. Schneider

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Introduction

Exposure to low levels of lead (Pb) impairs a variety of cognitive processes. Although children exposed to Pb developmentally present with a variety of cognitive impairments that include deficits in learning, memory, language, and executive functioning, experimental work on Pb toxicity in rats has focused mostly on learning and memory deficits and less on executive functions. However, detrimental effects on executive functioning could lead to or even underlie a variety of other cognitive problems attributed to Pb exposure. In this study, we examined the ability of Long Evans rats (control and Pb-exposed: 150ppm Pb-acetate in food given perinatally (gestation …

One-Step Generation Of Multiple Transgenic Mouse Lines Using An Improved Pronuclear Injection-Based Targeted Transgenesis (I-Pitt)., Masato Ohtsuka, Hiromi Miura, Keiji Mochida, Michiko Hirose, Ayumi Hasegawa, Atsuo Ogura, Ryuta Mizutani, Minoru Kimura, Ayako Isotani, Masahito Ikawa, Masahiro Sato, Channabasavaiah B. Gurumurthy

Journal Articles: Genetics, Cell Biology & Anatomy

BACKGROUND: The pronuclear injection (PI) is the simplest and widely used method to generate transgenic (Tg) mice. Unfortunately, PI-based Tg mice show uncertain transgene expression due to random transgene insertion in the genome, usually with multiple copies. Thus, typically at least three or more Tg lines are produced by injecting over 200 zygotes and the best line/s among them are selected through laborious screening steps. Recently, we developed technologies using Cre-loxP system that allow targeted insertion of single-copy transgene into a predetermined locus through PI. We termed the method as PI-based Targeted Transgenesis (PITT). A similar method using PhiC31-attP/B system …

Insertion Of Sequences At The Original Provirus Integration Site Of Mouse Rosa26 Locus Using The Crispr/Cas9 System., Rolen M. Quadros, Donald W. Harms, Masato Ohtsuka, Channabasavaiah B. Gurumurthy

Journal Articles: Genetics, Cell Biology & Anatomy

Targeted transgenic mouse models, where an exogenous gene is inserted into a specified genomic locus to achieve its stable and reliable expression, have been widely used in biomedical research. However, the available methodologies for targeted insertion of sequences require many laborious steps that involve the use of embryonic stem (ES) cells. We recently developed Pronuclear Injection-based Targeted Transgenesis (PITT), a method that uses a recombinase-mediated cassette exchange (RMCE) to enable insertion of sequences at a predetermined genomic locus, such as ROSA26. The PITT technique uses fertilized eggs (instead of ES cells) collected from 'seed mice' that contain the RMCE landing …

Generation Of A Retinoblastoma (Rb)1-Inducible Dominant-Negative (Dn) Mouse Model., Shikha Tarang, Songila M.S.R Doi, Channabasavaiah B. Gurumurthy, Donald W. Harms, Rolen M. Quadros, Sonia M. Rocha-Sanchez

Journal Articles: Genetics, Cell Biology & Anatomy

Retinoblastoma 1 (Rb1) is an essential gene regulating cellular proliferation, differentiation, and homeostasis. To exert these functions, Rb1 is recruited and physically interacts with a growing variety of signaling pathways. While Rb1 does not appear to be ubiquitously expressed, its expression has been confirmed in a variety of hematopoietic and neuronal-derived cells, including the inner ear hair cells (HCs). Studies in transgenic mice demonstrate that complete germline or conditional Rb1 deletion leads to abnormal cell proliferation, followed by massive apoptosis; making it difficult to fully address Rb1's biochemical activities. To overcome these limitations, we developed a tetracycline-inducible TetO-CB-myc6-Rb1 (CBRb) mouse …

Improved Accuracy Of Surgical Pathology Accessioning, Coding, And Billing Practices After Self-Evaluation, Allison F. Goldberg, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

INTRODUCTION

Accurate accessioning, coding, and billing of specimens are vital, but often disregarded, aspects of a surgical pathologist’s job. This study had two aims related to those aspects. First, to evaluate the quality of our systems based practices in order to identify areas for improvement. Second, to assess if changes made based on our self-evaluation improved accuracy.

Tumor Metabolism In The Microenvironment Of Nodal Metastasis In Head And Neck Squamous Cell Carcinoma, Mehri Mollaee, Md, Patrick T. Tassone, Md, David Cognetti, Md, Adam J. Luginbuhl, Md, Joanie Johnson, Ubaldo E. Martinez-Outshoorn, Md, Tingting Zhan, Joseph M. Curry, Md, Madalina Tuluc, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

ABSTRACT

Introduction: Monocarboxylate transporter 4 (MCT4) is a cell membrane transporter of lactate. MCT4 is a tumor-specific marker of oxidative stress, glycolysis and hypoxia in tumor stromal cells. We investigated HPV positive and negative tumors with regional metastases to cervical lymph nodes (LN) to study how the metastatic tumor cells interact with their microenvironment. By selecting cancers with extracapsular extension (ECE), we intended to evaluate the interaction between metastases and the surrounding extranodal tissue.

Methods:

Clinical data were collected from 24 advanced stage oropharyngeal squamous cell carcinoma (OPSCC) patients with neck LN metastasis. All patients presented with at least N1 …

Diagnostic Value Of Endoscopic Ultrasound-Guided Fine Needle Aspiration Of Intra-Abdominal Lymph Nodes In Patients With Concurrent Biopsy Of Intra-Abdominal Tumors, Allison F. Goldberg, Md, Charalambos C. Solomides, Md, Marluce Bibbo, Md, Rossitza Draganova-Tacheva, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

BACKGROUND

• Diagnostic endoscopic biopsy of intra-abdominal lesions may be performed prior to surgical resections.

• Endoscopic ultrasound-guided fine needle aspiration of intra-abdominal lymph nodes may also be performed to:

• Yield more information

• Allow for more accurate staging

• We evaluate the additional staging information concurrent fine needle aspiration of intra-abdominal lymph nodes provided

DESIGN

• We included all patients at our institution from January 1, 2000 to March 30, 2015 who, during the same endoscopic procedure, had:

• Endoscopic ultrasound guided fine needle aspiration of an intra-abdominal lymph node AND

• Endoscopic ultrasound guided fine needle aspiration …

Mcm2 And Chromogranin Are Markers Of Serrated Polyp Progression, Danielle Fortuna, Md, Bruce M. Boman, Md, Phd, Juan P. Palazzo, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

Objectives:

Using immunohistochemistry for MCMs and CGA:

Examine the proliferative compartment of SPs

Assess neuroendocrine cell population in SPs

Goal: Identify potential trends in the proliferative and the neuroendocrine cell compartments in SP progression compared to the background normal mucosa

Analysis Of Patient Results Distributions To Reevaluate A Reference Range Change For Calcium, After A Change In Assay Reagents On The Roche Cobas C500 Analyzer, Upasana Joneja, Md, Laura J. Mccloskey, Md, Douglas F. Stickle, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

Introduction:

A change in reagents for calcium (Ca) on the Roche Cobas c500 used in our laboratory analyzer took place in 2013. The previous reference range (8.5-10.5 mg/dL) was replaced with that from the manufacturer's study (8.6-10.0 mg/dL), based on correlation of results between the new and old assays. As a matter of quality assurance, we undertook a post-assay-change reevaluation of the reference range change, using a method based on that of Bhattacharya [1]. In short, the method relies on the assumption that the reference range is a normal distribution, which assumption enables this distribution to be isolated mathematically from …

Is Myeloproliferative Neoplasm With Splanchnic Vein Thrombosis A Distinct Clinical Entity?, Upasana Joneja, Md, Jerald Z. Gong, Md, Guldeep Uppal, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

Clinical History:

A 29 year-old previously healthy female presented with sub-acute symptoms of weight loss, right upper quadrant pain and nausea.

CBC results: WBC-11.7, Hb- 12.5, Platelet- 286, MCV- 90, MCV- 26.8, RDW- 18%.

Imaging: Ultrasound and MRI of the abdomen were suggestive of Budd-Chiari syndrome that was supported by a liver biopsy showing features of hepatic outflow obstruction(Figure 1). There was no clinical or radiological evidence of splenomegaly. The hypercoagulable work-up was negative. An underlying Myeloproliferative Neoplasm (MPN) was suspected.

Stromal Monocarboxylate Transporter Mct4 Is A Poor Prognostic Factor In Squamous Cell Carcinoma, Mehri Mollaee, Md, P. Cotzia, Patrick Tassone, Md, Adam J. Luginbuhl, Md, Ubaldo E. Martinez-Outshoorn, Md, Tingting Zhan, Joseph M. Curry, Md, Madalina Tuluc, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

ABSTRACT

Introduction:

Monocarboxylate transporter 4 (MCT4) is the main exporter of lactate out of cells. It is also a critical component in the glycolytic metabolism of cancer cells. In this study, stromal MCT4 in oral SCC was correlated with risk of recurrence (ROR), extent of primary tumor (pT) and nodal metastasis (pN), perineural invasion (PNI), lymphovascular invasion (LVI), HPV status, extracapsular extension (ECE) and positive margin.

Methods:

Clinical data were collected for 86 consecutive patients with oral HNSCC. Tissue microarrays (TMA) were constructed from paraffin blocks of resection specimens and stained for MCT4. Immunohistochemistry (IHC) staining was assessed and quantified …

The Predictive Value Of The Proliferation Marker Ki-67 In Patients With Fulminant Hepatic Failure, Ashesh P. Shah, Md, Upasana Joneja, Md, David Walls, Md, John Farber, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

INTRODUCTION

In the United States, acute liver failure (ALF) affects an estimated 2,000 people per year and accounts for 6% of all liver transplants. Without transplantation, however, less than 50% of patients survive with medical management alone. Early identification of patients with survivable AFL is important to guiding their management and early referral to transplantation.

Studies have shown that the current prognostic scoring systems used for FHF, including the King’s College Criteria (KCC) and the Model for End- Stage Liver Disease (MELD), have poor sensitivity and negative predictive value for predicting outcome, highlighting a need for more accurate predictive models. …

Assessment Of Utility Of Daily Patient Results Averages As Adjunct Quality Control In A Weekday-Only Satellite Chemistry Laboratory, Dimath Alyemni, Md, Laura J. Mccloskey, Md, Barbara Goldsmith, Phd, Facb, Aryana Treweek, Robert T. Stapp, Do, Douglas F. Stickle, Phd

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

ABSTRACT

Background:

Our department operates a weekday-only (8AM-5PM) satellite laboratory in an infusion center with a menu of 18 chemistry tests on a Roche c501 analyzer. We examined whether daily patient results averages (PRA) in this setting might be useful as a patient-based quality control (PBQC) adjunct to standard daily liquid quality control (LQC) measurements. First, we evaluated the reproducibility (coefficient of variation, CV) of daily PRAs for each analyte, and compared these to CVs of LQC. Second, for select analytes found to have relatively low PRA CVs, we evaluated the extent to which use of daily PRA measurements could …

Human Parechovirus And Enterovirus Initiate Divergent Innate Immune Responses In The Cns: Pathogenic And Diagnostic Implications, Danielle Fortuna, Md, Ana Maria Cardenas, Phd, Erin H. Graf, Phd, Larry A. Harshyne, Kevin Quann, Bs, D. Craig Hooper, Phd, Mark T. Curtis, Md, Phd

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

The picornaviruses human parechovirus (HPeV) and enterovirus (EV) cause a wide range of diseases, including CNS infections, which can be severe and potentially fatal. EV causes most cases of pediatric meningoencephalitis worldwide, and HPeV type 3 (HPeV3) is the most common cause of viral meningitis in young infants. Each year in the United States, there are over 75,000 cases of aseptic meningitis. Despite reassuring short-term outcomes, negative neurodevelopmental sequalae are increasingly associated with HPeV and EV.

The pathogenesis and severity of HPeV and EV infections are undoubtedly linked to the innate and adaptive immune responses elicited by these viruses. Until …

Quantification Of Csf Chemokines And Cytokines Allows For Rapid Laboratory Detection Of Cns Infections And Further Discrimination Between Viral And Non-Viral Pathogens, Danielle Fortuna, Md, Am Cardenas, Eh Graf, Amity L. Roberts, Phd, D(Abmm), Larry A. Harshyne, Phd, D. Craig Hooper, Phd, Mark T. Curtis, Md, Phd

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

Background:

Prompt diagnosis of central nervous system (CNS) disease is critical to guide intervention and appropriate therapy. Development of novel laboratory approaches to rapidly classify acute-onset CNS disease is in great demand. Serious microbial pathogens, especially viruses, are quickly expanding beyond their historic geographic range and may not even be considered in the clinician’s differential diagnosis. Unlike bacterial cultures, current viral testing targets a limited number of viruses. Additionally, despite diversity in etiology, signs and symptoms of both infectious and non-infectious CNS disorders can be remarkably similar, which can confuse the clinical picture and delay treatment. Bacterial, viral, fungal and …

Is Hemoglobin Variant Analysis Helpful In The Diagnostic Work-Up Of Patients Revealing Microcytic Erythrocytosis On Complete Blood Count?, Upasana Joneja, Md, Alina Dulau-Florea, Gene Gulati, Ph.D, Jerald Z. Gong, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

Introduction:

Microcytic erythrocytosis is an abnormal CBC (complete blood count) finding that is under-recognized, poorly understood, and consequently under-utilized in patient care. It is characterized by decreased MCV and increased RBC count. Its etiology is likely multifactorial and includes thalassemias and hemoglobinopathies. The focus of our study was to determine the relative prevalence of hemoglobin-associated disorders in patients revealing microcytic erythrocytosis on CBC and to demonstrate whether or not hemoglobin variant analysis should be included in the diagnostic work-up of such cases.

A Distinct Pattern Of Beclin-1 Staining Helps Distinguish Sessile Serrated Adenomas, Danielle Fortuna, Md, Juan P. Palazzo, Md

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

Autophagy, a lysosomal degradation system, has both cell survival and cell death-promoting capabilities, and its versatility is exploited in many pathologic entities. In neoplastic processes, autophagy has been demonstrated to contribute to both tumor suppression and tumorigenesis in a relatively tumor-specific fashion. Beclin-1 is a protein involved in the formation of the autophagosome, the core unit of autophagy, and serves as one of the general markers for this process. Previous studies have shown Beclin-1 overexpression in both pre-neoplastic and invasive colon carcinoma but weak to absent expression in normal colonic mucosa.

Serrated polyps (SPs) of the colon represent morphologically and …

Hiv Genotyping Cost Analysis With Follow-Up As An Indicator, Laura Biederman, Md, Amity L. Roberts, Phd, D(Abmm)

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

HIV-1 genotype (GHIV), HIV-1 Integrase (HIV1I) and HIV-1 Trophile (HIV1T) assays are sendout tests that incur a significant financial burden on the laboratory when ordered on inpatients who do not receive follow-up clinic visits. For these assays to be utilized in guiding antiretroviral therapy, the patient must receive follow-up. It will reduce the sendout budget by restricting these tests to the outpatient clinic setting.

Cerebrospinal Fluid Cytokine And Chemokine Patterns In Central Nervous System Infections, Hemorrhage And Neoplasms, Danielle Fortuna, Md, Larry A. Harshyne, Phd, D. Craig Hooper, Phd, Amity L. Roberts, Phd, D(Abmm), Danielle Hutchings

Department of Pathology, Anatomy, and Cell Biology Resident's Posters

Cytokines and chemokines are soluble proteins that act as regulators of cellular functions throughout the body. Cytokines and chemokines released in the setting of various CNS disorders appear in the CSF compartment where determination of their levels can provide insight into pathogenic processes such as neuroinflammation. We utilized the Millipore HCYTOMAG 60K assay/kit/system to perform multiplex profiling of 42 different cytokines/chemokines in the CSF of patients with a variety of distinct CNS disease processes, including infection, hemorrhage and neoplasia. CNS infections included viral (Chronic Parechovirus type 3 (HPeV3), Enterovirus (EV) 68, Adenovirus, JC virus, West Nile virus), bacterial (Mycobacterium tuberculosis, …

Validation Of Simple Sequence Length Polymorphism Regions Of Commonly Used Mouse Strains For Marker Assisted Speed Congenics Screening., Channabasavaiah B. Gurumurthy, Poonam S. Joshi, Scott G. Kurz, Masato Ohtsuka, Rolen M. Quadros, Donald W. Harms, K.C. Kent Lloyd

Journal Articles: Genetics, Cell Biology & Anatomy

Marker assisted speed congenics technique is commonly used to facilitate backcrossing of mouse strains in nearly half the time it normally takes otherwise. Traditionally, the technique is performed by analyzing PCR amplified regions of simple sequence length polymorphism (SSLP) markers between the recipient and donor strains: offspring with the highest number of markers showing the recipient genome across all chromosomes is chosen for the next generation. Although there are well-defined panels of SSLP makers established between certain pairs of mice strains, they are incomplete for most strains. The availability of well-established marker sets for speed congenic screens would enable the …

Assessment Of Artificial Mirna Architectures For Higher Knockdown Efficiencies Without The Undesired Effects In Mice., Hiromi Miura, Hidetoshi Inoko, Masafumi Tanaka, Hirofumi Nakaoka, Minoru Kimura, Channabasavaiah B. Gurumurthy, Masahiro Sato, Masato Ohtsuka

Journal Articles: Genetics, Cell Biology & Anatomy

RNAi-based strategies have been used for hypomorphic analyses. However, there are technical challenges to achieve robust, reproducible knockdown effect. Here we examined the artificial microRNA (amiRNA) architectures that could provide higher knockdown efficiencies. Using transient and stable transfection assays in cells, we found that simple amiRNA-expression cassettes, that did not contain a marker gene (-MG), displayed higher amiRNA expression and more efficient knockdown than those that contained a marker gene (+MG). Further, we tested this phenomenon in vivo, by analyzing amiRNA-expressing mice that were produced by the pronuclear injection-based targeted transgenesis (PITT) method. While we observed significant silencing of the …

De Novo Assembly Of The Chimpanzee Transcriptome From Nextgen Mrna Sequences, Mnirnal D. Maudhoo, Jacob D. Madison, Robert B. Norgren

Journal Articles: Genetics, Cell Biology & Anatomy

BACKGROUND: Common chimpanzees (Pan troglodytes) and bonobos (Pan paniscus) are the species most closely related to humans. For this reason, it is especially important to have complete and accurate chimpanzee nucleotide and protein sequences to understand how humans evolved their unique capabilities. We provide transcriptome data from four untransformed cell types derived from the reference Pan troglodytes, "Clint", to better annotate the chimpanzee genome and provide empirical validation for proposed gene models of this important species.

FINDINGS: RNA was extracted from primary cells cultured from four tissues: skin, adipose stroma, vascular smooth muscle and skeletal muscle. These four RNA samples …